report acceptance: 2020 ltemp sampling & mussel

Report Acceptance: 2020 LTEMP Sampling & Mussel Transcriptomics 4-12

951.104.210406.4-12LTEMPRPT

Briefing for PWSRCAC Board of Directors – May 2021

ACTION ITEM

Sponsor: Austin Love and the Scientific Advisory Committee

Project number and name or topic: 9510 – Long-Term Environmental Monitoring Program

1. Description of agenda item: This agenda item seeks Board approval for two reports.The first report is titled “Long-Term Environmental Monitoring Program: 2020 SamplingResults and Interpretations” by James R. Payne, of Payne Environmental ConsultantsIncorporated, and William B. Driskell. This report is produced annually to provide an analyticalsummary and interpretation of the passive sampling device, mussel, and sediment samplestaken each summer as part of the Council’s environmental monitoring work. Those samples arecollected and analyzed in order to monitor hydrocarbon (oil) contamination associated with theoperation of the Valdez Marine Terminal and associated tankers. The report goes into greatdetail regarding the results of the 2020, and historic field sampling and lab analyses, but theExecutive Summary, Introduction, Summary Points, and Conclusions sections should providethe average reader with enough information to understand the most important aspects of thiswork.

This agenda item also seeks Board approval for the report titled “Using Mussel Transcriptomics for Environmental Monitoring in Port Valdez, Alaska: 2019 and 2020 Pilot Study Results” by Lizabeth Bowen, Austin Love, Shannon Waters, Katrina Counihan, Brenda Ballachey, Heather Coletti, William B. Driskell, and James R. Payne. This report summarizes the results of a two-year pilot study investigating the utility of using mussel transcriptomics as part of the Council’s Long-Term Environmental Monitoring Program. Transcriptomics is an environmental monitoring technique in which the genes of an organism are analyzed to understand how environmental stressors, such as oil pollution or temperature, are impacting an organism – in this case blue mussels. This is a very technical report meant to be used by the Scientific Advisory Committee to decide if transcriptomics should continue to be used as part of the Council’s Long-Term Environmental Monitoring Program.

A summary presentation of the results from the two reports will be provided by Austin Love and the authors of the reports, James Payne, William Driskell, and Lizabeth Bowen, will be available to address comments and questions regarding their work.

2. Why is this item important to PWSRCAC: The Long-Term Environmental MonitoringProgram helps PWSRCAC fulfill one of its duties detailed in the Oil Pollution Act of 1990. TheAct instructs the PWSRCAC to “devise and manage a comprehensive program of monitoring theenvironmental impacts of the operations of terminal facilities and of crude oil tankers whileoperating in Prince William Sound.” The work done under the Council’s Long-TermEnvironmental Monitoring Program has been designed by the Scientific Advisory Committee toachieve that Act mandate.


951.104.210406.4-12LTEMPRPT

3. Previous actions taken by the Board on this item: The Long-Term Environmental Monitoring Program has been conducted by PWSRCAC since 1993, and many actions have been taken by the Board on this item since that time. In the interest of providing currently pertinent information regarding actions items taken by the Board on this item, only the last five years of actions are presented below. However, all historic actions pertaining to this agenda item are available for review upon request (please contact Austin Love for that information). Meeting Date Action Board 9/17/2015 The Board accepted and approved the 2008-2013 Combined Annual LTEMP Report by

contractor Payne Environmental Consultants titled “Long Term Environmental Monitoring Program Results and Interpretations from Sampling, 2008-2013.”

Board 5/5/2016 The Board approved the following: 1. Contractor selection and contracting with Payne Environmental Consulting, Inc., for sampling, training, and analytical reporting work on mussels and sediments to be performed under the Long-Term Environmental Monitoring Project (LTEMP) for FY2017 at an amount not to exceed $52,390. 2. Contractor selection and contracting with NewFields-Environmental Forensics Practice for analytical laboratory work to be performed under LTEMP for FY2017 at an amount not to exceed $28,625. 3. Contractor selection and contracting with Oregon State University for analytical laboratory work on passive sampling devices to be performed under LTEMP for FY2017 at an amount not to exceed $27,750.

Board 5/4/2017 The Board approved: (1) Contractor selection and contract negotiation with Payne Environmental Consulting Inc. for sampling, training and analytical reporting work on mussels and sediments to be performed under the LTEMP for FY18 at an amount not to exceed $45,960. (2) Contractor selection and negotiation with NewFields Environmental Forensics Practice for analytical laboratory work and sample storage to be performed under LTEMP for FY18 at an amount not to exceed $51,592. (3) Contractor selection and contract negotiation with Oregon State University for analytical laboratory work on passive sampling devices to be performed under LTEMP for FY18 at an amount not to exceed $27,750 and (4) The 2017 LTEMP Report “Long-Term Environmental Monitoring Program Final Report: 2015 Sampling Results and Interpretations” as approved by SAC at its January 31, 2017 meeting.

XCOM 6/7/2017 The Executive Committee authorized a change order to contract number 951.17.03 with NewFields Companies, LLC by adding $2,619 to the contract. This would increase their contract from the current amount of $41,300 to $43,919.

Board 1/18/2018 The Board accepted the report titled “Long-Term Environmental Monitoring Program - Final Report: 2016 Sampling Results and Interpretations” (prepared by James R. Payne Ph.D. and William B. Driskell) as meeting the terms and conditions of the contract and for posting on the PWSRCAC website.

Board 5/3/2018 The Board approved: contract with Payne Environmental (PECI) for sampling and analytical reporting work on mussels and sediments to be performed under FY19 LTEMP not to exceed $139,086; contract with NewFields Companies, LLC for analytical laboratory work and sample storage under LTEMP FY19 not to exceed $61,402; contract with Oregon State University (OSU) for passive sample device purchase and analytical lab work on passive sampling devices under LTEMP for FY19 not to exceed $27,310; and authorized this contract work to commence prior to the start of FY19 to accommodate tidal considerations in an estimated amount of $20,000.

Board 5/3/2018 The Board accepted the report titled “September 2017 Berth 5 Oil Spill - Sampling Results and Interpretations” by James R. Payne, Ph.D., and William Driskell for distribution & posting on PWSRCAC’s website.

Board 5/2/2019 The Board authorized contract negotiations with Payne Environmental Consultants for sampling and analytical report work on mussels and sediments to be performed under LTEMP for FY20, at an amount not to exceed $65,866; and authorized contract negotiations with Newfields Environmental Forensics Practice for analytical laboratory work and sample storage to be performed under LTEMP for FY20 at an amount not to


951.104.210406.4-12LTEMPRPT

exceed $28,506. Authorized contract negotiations with Oregon State University for passive sample device purchase and analytical laboratory work on passive sampling devices to be performed under LTEMP for FY20, at an amount not to exceed $20,590; and authorized contract work to commence prior to the start of FY20, as approximately $20,000 of these funds will need to be expended in May and June 2019 because of the supply prerequisites and sampling timing.

Board 9/19/2019 The Board accepted the report titled “Long Term Environmental Monitoring Program: 2018 Sampling Results and Interpretations” by Dr. James R. Payne and William B. Driskell, dated July 2019 as meeting the terms of the contract and for distribution.

Board 1/23/2020 The Board accepted of the “Port Valdez Mussel Transcriptomics” report by Lizabeth

Bowen of the U.S. Geological Survey, dated November 20, 2019, as meeting the terms and conditions of contract number 951.20.06, and for distribution to the public.

Board 5/7/2020 The Board accepted the report titled “Long-Term Environmental Program: 2019 Sampling Results and Interpretations,” by Dr. James Payne and William B. Driskell, dated March 2020, as meeting the terms and conditions of contract number 951.20.04, and for distribution to the public.

Board 5/21/2020 Approval of FY2021 Contracts for Project 9510 LTEMP - The Board approved the following: Authorizing a contract negotiation with Payne Environmental Consultants Inc., for work to be performed under LTEMP, at an amount not to exceed $115,064. Authorizing a contract negotiation with Newfields Environmental Forensics Practice, for work to be performed under LTEMP, at an amount not to exceed $95,807. Authorizing a contract negotiation with the United States Geological Survey, for work to be performed under LTEMP, at an amount not to exceed $65,371. Authorizing a contract negotiation with Oregon State University, for work to be performed under LTEMP, at an amount not to exceed $22,030. Authorizing a contract work to commence prior to the start of FY2021, as approximately $33,000 of these funds will need to be expended in May and June 2020.

4. Summary of policy, issues, support or opposition: Regarding the transcriptomics report, this is a pilot study project that is being considered for regular inclusion in the Council’s Long-Term Environmental Monitoring Program. The reasons why this is considered a pilot study project are because transcriptomics monitoring represents a cutting edge method to conduct environmental monitoring and the Council had never used this technique before. The Scientific Advisory Committee wanted to conduct this pilot work for two years before deciding if a recommendation should be made to include this as a permanent part of Long-Term Environmental Monitoring Program. The results of the 2019 and 2020 work have been promising, supporting the continued use of mussel transcriptomics monitoring. 5. Committee Recommendation: The Scientific Advisory Committee recommends that the Board of Directors accept both reports and make them available for public distribution. 6. Relationship to LRP and Budget: Project 951 – Long Term Environmental Monitoring is in the approved FY2021 budget and annual work plan.

9510--Long Term Environmental Monitoring As of March 22, 2021

FY-2021 Budget Original $310,947.00 Modifications ($6,000.00)


951.104.210406.4-12LTEMPRPT

Revised Budget $304,947.00

Actual and Commitments Actual Year-to-Date $233,879.18 Commitments (Professional Services) $9,194.00 Actual + Commitments $243,073.18

Amount Remaining $61,873.82

7. Action Requested of the Board of Directors:• Accept the report titled “Long-Term Environmental Monitoring Program: 2020

Sampling Results and Interpretations,” by Dr. James R. Payne and William B. Driskell,dated March 2021, as meeting the terms and conditions of contract number 951.21.04,and for distribution to the public.

• Accept the report titled “Using Mussel Transcriptomics for Environmental Monitoringin Port Valdez, Alaska: 2019 and 2020 Pilot Study Results”, dated February 17, 2021, asmeeting the terms and conditions of contract number 951.21.06 and for distribution tothe public.

8. Alternatives: Do not accept the reports or accept the reports with recommendedrevisions.

9. Attachments:A: Draft report titled “Long-Term Environmental Monitoring Program: 2020 Sampling Resultsand Interpretations” by James R. Payne and William B. Driskell.

B: Draft report titled “Using Mussel Transcriptomics for Environmental Monitoring in Port Valdez, Alaska: 2019 and 2020 Pilot Study Results” by Lizabeth Bowen, Austin Love, Shannon Waters, Katrina Counihan, Brenda Ballachey, Heather Coletti, William B. Driskell, and James R. Payne.

LONG-TERM ENVIRONMENTAL MONITORING PROGRAM:2020 SAMPLING RESULTS AND INTERPRETATIONS

JAMES R. PAYNE, PH.D. PAYNE ENVIRONMENTAL CONSULTANTS, INC. ENCINITAS, CA

WILLIAM B. DRISKELL INDEPENDENT CONSULTANT SEATTLE, WA

APRIL 2021

PWSRCAC Contract No.

951.21.04

The opinions expressed in this commissioned

report are not necessarily those of PWSRCAC.

4-12 Attachment A

2020 LTEMP Report

ii

CONTENTS List of Figures ................................................................................................................................................................ iii

List of Tables ................................................................................................................................................................. vi

Abbreviations .............................................................................................................................................................. viii

Executive Summary ....................................................................................................................................................... x

Introduction ................................................................................................................................................................... 1

Valdez Marine Terminal BWTF Effluent .................................................................................................................... 5

Port Valdez Sediments .............................................................................................................................................. 9

Sediment TPAH Trends 1993-2020 ....................................................................................................................... 9

Sediment Biomarkers .......................................................................................................................................... 11

Valdez Marine Terminal Sediments .................................................................................................................... 11

Gold Creek Sediments ......................................................................................................................................... 15

Port Valdez Mussel Tissues ..................................................................................................................................... 20

Historical Trends in Port Valdez Mussel Tissues ................................................................................................. 20

Non-Spill-Related Patterns in Port Valdez Mussels ............................................................................................ 22

Valdez Marine Terminal Historical Mussel Patterns, Pre-2020 Spill ................................................................... 23

Jackson Point Mussels ......................................................................................................................................... 26

Gold Creek Mussels ............................................................................................................................................. 30

Supplemental Mussel Stations in 2020 ................................................................................................................... 32

Greater PWS and GOA Stations ............................................................................................................................... 33

Passive Sampling Devices ........................................................................................................................................ 34

Related Topics ............................................................................................................................................................. 38

Summary Points ........................................................................................................................................................... 40

Conclusions .................................................................................................................................................................. 42

Sediments ........................................................................................................................................................... 42

Tissues ................................................................................................................................................................. 42

Passive Samplers ................................................................................................................................................. 42

Acknowledgements ..................................................................................................................................................... 43

References ................................................................................................................................................................... 43

Appendix 1. Polycyclic Aromatic Hydrocarbon (PAH), Saturated Hydrocarbon (SHC), And Biomarker Analytes ........ 51

Appendix 2. Analytic Results for 2020 Field Samples and Blanks ................................................................................ 55

Appendix 3. Project History ......................................................................................................................................... 66

Appendix 4. Methods ................................................................................................................................................. 70

4-12 Attachment A

2020 LTEMP Report

iii

Data analysis ........................................................................................................................................................... 72

Biomarkers .............................................................................................................................................................. 73

Appendix 5. Laboratory Performance ......................................................................................................................... 74

Laboratory Quality Control ................................................................................................................................. 74

Method Detection Limits .................................................................................................................................... 74

Appendix 6. Sediment Grain Size ................................................................................................................................. 76

Appendix 7. Sediment TOC .......................................................................................................................................... 80

Appendix 8. Oxygenated Products in Treated Ballast Water Discharges ................................................................... 82

Appendix 9. Related Topics......................................................................................................................................... 84

Appendix 10. Beyond LTEMP ....................................................................................................................................... 87

Cover image – “Staging at Berth 5” Photo by William Driskell.

LIST OF FIGURES Figure 1. LTEMP sampling stations in Port Valdez adjacent to the terminal (AMT-B, AMT-S, and JAP) and 6 km NW of

the VMT (GOC). New stations in 2020 include Red and Green sites in the Valdez Harbor entrance plus Jack

and Galena Bay in Valdez Arm. This Google Earth image (June 2019) shows a tanker docked at Berth 5. ...... 2

Figure 2. Map of the LTEMP sites with station abbreviations. PWS and GOA sites are sampled every five years,

whereas Port Valdez sites are sampled annually. ............................................................................................. 3

Figure 3. PAH and biomarker profiles (ng/L) of raw (unfiltered) BWTF-BTT effluent samples from July 2016 (upper

plots) and March 2017 (lower plots). The dotted red line represents an overlay of fresh ANS crude oil

normalized (scaled) to hopane (T19, colored gold in the biomarker profiles). Excess dissolved PAH

constituents are observed in 2017 as N-, F- and DBT-group analytes above the source reference line (lower

left). ................................................................................................................................................................... 5

Figure 4. PAH, SHC, and biomarker profiles (ng/L) of the filtered, particulate/oil-phase droplets in the BWTF-BTT

effluent. The red line overlay represents fresh ANS scaled to the sample’s hopane (T19, colored gold in the

biomarker plots). The gaps between the measured PAH and the overlay portray the extent of weathering.

The TPAH concentrations are similar (1,639 and 2,083 ng/L) but there is additional loss of the higher-

molecular-weight (FPs, NBTs, and Cs) in the summer due to enhanced biodegradation and longer residence

time in the BTT. Biomarkers show essentially no degradation in both seasons. SHC (middle plots) show losses

of lower-molecular-weight C9 through C15 components in fresh ANS crude oil largely due to evaporation. In

summer SHC, C32 was a matrix interferent; SHC are scaled to C27. ................................................................... 7

Figure 5. PAH profiles of BTT effluent samples collected under summer (July 2016) and late winter (March 2017)

conditions: A) whole unfiltered sample; B) particulate/oil phase trapped on the glass-fiber-filter; and C)

dissolved phase. The dotted red lines represent fresh ANS crude oil PAH profile normalized to hopane to

show evaporation and dissolution effects on lower-molecular-weight PAH (C profiles lack the indissolvable

hopane for scaling). Note, however, that while the dissolved-phase patterns are similar, the winter TPAH

concentrations are an order of magnitude higher. ........................................................................................... 8

Figure 6. Time series of log (TPAH43) in sediments at AMT-S and GOC-S, 1993-2020. .............................................. 10

Figure 7. PAH and biomarker profiles of a representative 2016 AMT-S sediment sample overlaid with fresh ANS crude

oil reference (dotted red lines) when alternatively scaled by the highly conservative biomarker, hopane

4-12 Attachment A

2020 LTEMP Report

iv

(upper plots), versus the less recalcitrant PAH, NBT2 (lower plots). The lower plot biomarkers exceeding the

reference profile (here pointing out hopane, colored gold) demonstrate an accumulated excess relative to

the more easily degraded, residual PAH in this sample, plus an accumulation of the more recalcitrant NBT

and chrysene PAH homologues. ..................................................................................................................... 11

Figure 8. PAH, SHC, and biomarker concentrations and profiles of 2020 AMT-S sediment replicates with mixed

background, petrogenic, and combustion sources. The dotted red line in the PAH and biomarker profiles is

the July 2016 particulate-phase BWTF reference (Figure 4 and Figure 5) normalized against the sample’s

hopane; SHC ref normalized to C27. Biomarkers confirm the presence of a weathered ANS profile in the PAH.

........................................................................................................................................................................ 13

Figure 9. Representative PAH and SHC signatures of AMT sediments between 2011 and 2020 showing the

progression from a primarily pyrogenic PAH signature to a mix of pyrogenic and water-washed petrogenic

components with increasing terrestrial biogenic SHC and decreasing higher-molecular-weight residual

petrogenic waxes through 2020. Note dominance of 2019 background naphthalenes (N-N4). .................... 14

Figure 10. Time-series AMT-S sediment PAH profiles from 2011 and 2017 through 2020 along with the biomarker

profiles obtained after introducing those analyses to LTEMP in 2011. The dotted red line denotes the PAH

and biomarker profiles from the July 2016 BWTF particulate-phase normalized to hopane (see Figure 4 top).

Biomarkers confirm ANS oil in all samples. Reporting of TAS biomarkers began in 2017, MAS biomarkers in

2018. ............................................................................................................................................................... 15

Figure 11. PAH, SHC, and biomarker profiles of 2020 GOC-S sediment replicates. The dotted red line is July 2016

BWTF particulate phase reference (Figure 4) normalized against the sample’s hopane; for SHC normalized to

C27 (Figure 4). The T32 biomarker spikes are laboratory artifacts. ................................................................. 17

Figure 12. Representative PAH and SHC from GOC-S sediments between 2011 and 2020 showing very similar,

background naphthalene components and pyrogenic, parent-dominated, PAH and higher-molecular-weight

combustion products. SHC patterns and concentrations reflect terrestrial (plant wax) biogenic inputs since

2011. Red dashed line is sample-specific MDL. .............................................................................................. 18

Figure 13. GOC-S sediment PAH and biomarker profiles between 2011 and 2020. The dotted red line is July 2016

BWTF particulate-phase reference. Analyses in 2011 by ABL did not include the triaromatic steroid

biomarkers (TAS); later included in the analyses by Alpha/NewFields. .......................................................... 19

Figure 14. Time series of mean mussel TPAH43 concentrations comparing 2020 AMT-B, JAP-B and GOC-B with prior

LTEMP collections at other regional sites (open circles). Note the log scale for TPAH concentrations. ......... 21

Figure 15. PAH and biomarker patterns at AMT-B relating to the tanker loading arm spill at the terminal in September

2017. The samples show the background profile from the normal LTEMP collections pre-spill (top, July 2017,

TPAH 7 ng/g), the weathered oil in the mussels ~one-week post spill (middle, 108 ng/g), and three months

post-spill (bottom, 10 ng/g). Dotted red lines represent fresh ANS crude oil profiles normalized to the

sample’s hopane. The below-red-line gaps (middle left panel) show loss (evaporation and dissolution

weathering) of lower-molecular-weight PAH. Biomarker T26 is a laboratory artifact. .................................. 22

Figure 16. Representative LTEMP Tissue results from June 2020 showing near- or below-MDL, mixed pyrogenic-

dominated PAH patterns and biogenic SHC at AMT, JAP, and GOC along with an associated laboratory

method blank. The dotted red line is the sample’s method detection limit. There is a slightly elevated (but

still below MDL) and more complex PAH pattern at AMT and JAP which may indicate traces of residual PAH

from the VMT spill in April 2020 but this is not reflected in either the SHC or biomarker patterns. ............. 23

Figure 17. Time-series mussel PAH and SHC profiles from AMT-B. In 2008, the patterns show primarily below-MDL

water-washed (possibly petrogenic) naphthalenes and combustion products (P/A and FL plus PY). In 2015,

there are above-MDL dissolved-phase naphthalenes and trace-level combustion-product PAHs. In both 2018

and 2019, most of the below-MDL components are also associated with the lab blank. In June 2020, all of

4-12 Attachment A

2020 LTEMP Report

v

the below MDL PAH are derived from combustion products. Planktonic biogenic SHC (n-C15, n-C17, and

pristane) are also present in most of the samples. The dotted red line denotes the sample-specific MDL. .. 25

Figure 18. Time-series PAH and SHC profiles of mussels collected at JAP in 2016 through 2020 showing primarily

trace-level dissolved- and (possibly) particulate-phase background naphthalenes plus combustion product

PAHs (Ph, FL, PY, C, BBF, BKE, BEP), and perylene (PER) in 2016 and 2017. The below-MDL PAH in 2018 are

suspected of being procedural artifacts associated with the laboratory method blanks). The dotted red line

denotes the sample-specific MDL. .................................................................................................................. 27

Figure 19. April 2020 VMT spilled oil, Hot Zone mussels (TPAH 271,000 ng/g DW), and Jackson Point mussels (TPAH

350 ng/g DW) collected approximately three weeks after the spill. TAS and higher-molecular-weight

biomarkers are measured in oil but are not available for mussel extracts (gap on right). The dotted red line

denotes the sample-specific MDL. .................................................................................................................. 28

Figure 20. Jackson Point Mussels histograms showing the June 2019 unoiled background pattern, the May 2020

petrogenic profile collected three weeks post spill, and the June 2020 LTEMP profile demonstrating the

recovery to a mixed petrogenic and biogenic pattern. The dotted red line denotes the sample-specific MDL.

........................................................................................................................................................................ 29

Figure 21. Time-series PAH and SHC profiles of GOC mussels collected between 2008 and 2020. The 2008 PAH suggest

possibly particulate-phase, water-washed petrogenic naphthalenes, fluorenes, and DBTs (red tents) plus

below-MDL combustion products. 2015 shows only above-MDL dissolved-phase naphthalenes (also

observed at AMT-B at that time). In 2018 (and 2019, not shown), only at- or below-MDL traces of dissolved

naphthalenes and combustion products are suggested but these same patterns were observed in the

laboratory method blanks. The 2020 PAH profiles are derived exclusively from combustion products. SHC in

all years are derived from marine phytoplankton and copepods. Dotted red line denotes the sample-specific

MDLs. .............................................................................................................................................................. 31

Figure 22. Supplemental stations at Galena Bay, Jack Bay, and Valdez Small Boat Harbor entrance (Red and Green

navigation lights). Image from Google Earth dated 12/2016. ......................................................................... 32

Figure 23. Representative PAH, SHC, and S/T patterns from the four new stations Red and Green (below the red and

green channel navigation lights on the Valdez Small Boat Harbor entrance breakwater) and Jack Bay and

Galena Bay. The dotted red line represents ANS oil from the VMT spill incident. ........................................ 33

Figure 24. Passive sampling devices (PSD) consisting of a low-density polyethylene membrane enclosed in a stainless-

steel container and deployed subsurface in near-shore subtidal waters adjacent to LTEMP mussel collection

sites for up to 30 days prior to mussel sampling. Sampling photo courtesy of David Janka. ......................... 34

Figure 25. PAH profiles from 2018 and 2020 PSD deployments. The dominant naphthalenes (blue) are scaled to the

left axis and the two order-of-magnitude lower concentrations of other PAH (Fs. P/As, DBTs, and Cs) (red)

are scaled to the right axis of each plot. ......................................................................................................... 37

Figure 26. Example PAH profiles of dissolved-phase PAH, A) leaching from remarkably fresh residual Exxon Valdez oil

on Knight Island, 15 years post-spill (from Payne et al., 2005d); B) dissolved-phase BWTF effluent from March

2017. Descending naphthalene patterns are the compliment to ascending patterns in water-washed

particulate oil. ................................................................................................................................................. 38

Figure 27. Time series of various DMR parameters as reported in Alyeska’s monthly DMRs from October 2015

through January 2021. Red dots represent LTEMP’s June or July sampling events to demonstrate the variance

not captured by the “snapshot” mussel and sediment collection frequencies. ............................................. 39

Figure 28. LTEMP sampling stations in Port Valdez adjacent to (AMT-B, AMT-S, and JAP) and 6 km northwest (GOC)

of the VMT. This Google Earth image shows a tanker docked at Berth 5. ...................................................... 66

Figure 29. Map of the LTEMP sites with station abbreviations. .................................................................................. 67

Figure 30. Annual pipeline oil throughput (M barrels) from Alyeska statistics. .......................................................... 70

4-12 Attachment A

2020 LTEMP Report

vi

Figure 31. Deploying the Van Veen grab (upper left). View of benthic sediments collected with intact surface layer

(and residual water) in jaws (upper right) before sampling (bottom). Work photos courtesy of David Janka.

........................................................................................................................................................................ 71

Figure 32. Example of passive sampling device (PSD) consisting of a low-density polyethylene membrane strip

enclosed in a stainless-steel container and deployed subsurface in near-shore subtidal waters adjacent to

LTEMP mussel collection sites. Work photos courtesy of David Janka. .......................................................... 72

Figure 33. Individual replicate cumulative grain-size curves (%) for GOC-S and AMT-S, 2006-2020. 2020’s reps are

indicated by dotted red lines. ......................................................................................................................... 77

Figure 34. 3D plots of grain size components from GOC-S and AMT-S 2006-2020. Sampling years are color coded:

blue to orange, 2006-2019; Red, 2020. Note the clipped axes emphasize a decreasing shift in clay vs silt at

GOC-S in 2015. The two 2019 AMT-S outlier samples with anomalously high sand content (~20%) appear to

be sampling method errors. ............................................................................................................................ 79

Figure 35. LTEMP Total Organic Carbon trends in AMT-S and GOC-S sediments (% DW), 1993-2020........................ 80

Figure 36. Total organic carbon for sediments in Alyeska’s 2017 and 2018 monitoring program (from Shaw &

Blanchard, 2018, 2019) in vicinity of the terminal (near shallow sites) and deeper within the fjord (far deep

sites). ............................................................................................................................................................... 81

Figure 37. Iatroscan (TLC-FID) of ANS oil, BWTF raw effluent, and BWTF dissolved components from July 2016

showing relative abundance of single- and double-oxygenated (weathered) hydrocarbons relative to SHC and

PAH components. Courtesy of Christoph Aeppli. ........................................................................................... 83

Figure 38. PAH plot of shrimp eggs taken east of the VMT, which in our assessment, shows a water-washed,

weathered particulate-oil pattern absorbed through the chitin, lipid membrane and into the fat of the eggs.

From Carls et al., 2016. ................................................................................................................................... 84

Figure 39. Historic monthly stream discharge rates (cubic ft/sec) for Lowe River and Valdez Glacier Stream, 2015-

2019 (from USGS 2019). .................................................................................................................................. 86

Figure 40. Status and trends result from National Mussel Watch data (Kimbrough et al., 2008). All Alaska sites

characterized as low concentrations. ............................................................................................................. 89

Figure 41. Summary page of Alaska regional Mussel Watch results and trends based on 2004-05 report from

Kimbrough et al., 2008. ................................................................................................................................... 90

LIST OF TABLES Table 1. LTEMP tissue sampling history showing change in annual events coded for seasons. Spring, summer (SS);

spring, summer, autumn (SSA); or summer only (S). Sediments (not shown) have only been sampled in spring

and summer at AMT-S and GOC-S from 1993-2008, and afterwards only in summer. .................................... 4

Table 2. Ratios of n-C17/pristane and n-C18/phytane for July 2016 and March 2017 BTT effluent samples

(concentrations in ng/L). Lower ratios indicate extent of microbes preferentially degrading the alkanes over

the pristane and phytane isoprenoids. ............................................................................................................. 9

Table 3. Summary statistics for 2020 TPAH50 and TPAH43 concentrations (ng/g DW). ............................................. 10

Table 4. Historic average sediment TPAH43 values (ng/g DW), 2000-2020. ............................................................... 10

Table 5. Time series of mean TPAH43 (ng/g DW, n=3) from AMT-B, GOC-B, and JAP-B mussels, 2008-2020. ........... 21

Table 6. LTEMP tissue sampling history showing change in annual events coded for seasons. Spring, summer (SS);

spring, summer, autumn (SSA); or summer only (S). Sediments (not shown) were sampled in spring and

summer at AMT-S and GOC-S from 1993-2008, and afterwards only in summer. ......................................... 68

Table 7. Recent average daily throughput of Alyeska Pipeline and ballast water treatment (MGD). ......................... 70

Table 8. Surrogate recovery statistics by matrix from 2020 Alpha Laboratory analyses............................................. 74

4-12 Attachment A

2020 LTEMP Report

vii

Table 9. Alpha Analytical MDL target ranges. .............................................................................................................. 75

Table 10. Average grain size components for GOC-S and AMT-S, 2006-2020. ............................................................ 78

Table 11. LTEMP Total Organic Carbon in AMT-S and GOC-S sediments (% DW), 1993-2020. ................................... 81

Table 12. Most recent TPAH concentrations in LTEMP mussel tissues (ppb, ng/g DW) relative to 2004-2010 NOAA

Mussel Watch monitoring data and a recovered Alaska oil spill event. ......................................................... 88

4-12 Attachment A

2020 LTEMP Report

viii

ABBREVIATIONS Stations:

AMT-B Alyeska Pipeline Service Company’s Valdez Marine Terminal, Saw Island, Port Valdez AMT-S Alyeska Pipeline Service Company’s Valdez Marine Terminal, Berth 4, Port Valdez AIB Aialik Bay, west of Seward COH Constantine Harbor, Hinchinbrook Entrance, PWS (no longer sampled) DII Disk Island, Knight Island Group, western PWS GOC Gold Creek, Port Valdez JAP Jackson Point, Port Valdez KNH Knowles Head, eastern PWS SHB Sheep Bay, eastern PWS SHH Shuyak Harbor, Kodiak SLB Sleepy Bay, Latouche Island, western PWS WIB Windy Bay, outer Kenai Peninsula ZAB Zaikof Bay, Montague Island, central PWS *-B suffix code for biological (tissue) sample, e.g., AMT-B *-S suffix code for sediment sample, e.g., AMT-S

ABL Auke Bay Laboratory, NOAA/NMFS, Juneau, Alaska AHC Aliphatic hydrocarbons (same as saturated hydrocarbons – SHC) ANS Alaska North Slope APDES Alaska Pollutant Discharge Elimination System, successor to NPDES BTT Biological Treatment Tank BWTF Ballast Water Treatment Facility cm Centimeter DMR Discharge monitoring report DW Dry weight EMAP Environmental Mapping Project, EPA/Cook Inlet Regional Citizens Advisory Council EMP Environmental Monitoring Program, Alyeska Terminal EPA Environmental Protection Agency EVOS Exxon Valdez oil spill g Gram GC/FID Gas chromatography/flame ionization detector GC/MS Gas chromatography/mass spectrometry GERG Geochemical and Environmental Research Group, Texas A&M University GOA Gulf of Alaska GPS Global Positioning System KLI Kinnetic Laboratories, Inc., Anchorage, Alaska km Kilometers L Liter LTEMP Long-Term Environmental Monitoring Program m Meter MDL Analytic method detection limit MGD Million gallons per day mL Milliliter ng/g nanogram per gram NIST National Institute of Standards and Technology NMFS National Marine Fisheries Service NOAA National Oceanic and Atmospheric Administration NPDES National Pollutant Discharge Elimination System NRDA Natural Resource Damage Assessment OSU Oregon State University PAH Polycyclic (or polynuclear) aromatic hydrocarbons (listed in App. I)

4-12 Attachment A

2020 LTEMP Report

ix

PECI Payne Environmental Consultants, Inc., Encinitas, California PGS Particle grain size PSD Passive sampling device PW Produce waters PWS Prince William Sound PWSRCAC Prince William Sound Regional Citizens’ Advisory Council QC Quality control RL Reporting level SHC Saturated hydrocarbons (same as AHC: n-alkanes + pristane and phytane) (listed in App. I) SIM Selected ion monitoring SOP Standard operating procedure SQV Sediment quality values SRM Standard reference material, National Institute of Standards and Technology S/T Sterane/triterpane oil biomarkers (listed in App. I) TAS Triaromatic steroids TOC Total organic carbon TPAH Total PAH TSHC Total saturated hydrocarbons (same as total alkanes) UAF University of Alaska Fairbanks VMT Valdez Marine Terminal, Alyeska Pipeline Service Company NOTE: The abbreviation lists for PAH, SHC, and biomarker analytes can be found in Appendix 1.

4-12 Attachment A

2020 LTEMP Report

x

RESULTS AND INTERPRETATIONS FROM LTEMP SAMPLING, 2020

EXECUTIVE SUMMARY The Prince William Sound Regional Citizens’ Advisory Council (PWSRCAC) Long-Term Environmental Monitoring

Program (LTEMP) was begun in 1993, following the 1989 Exxon Valdez oil spill (EVOS), with the goal of monitoring

environmental impacts from oil transportation activities. To accomplish this task, the program has historically

sampled for oil-related contaminants in both mussel tissues and sediments in Port Valdez next to and across from

Alyeska’s Valdez Marine Terminal (VMT) in addition to sampling mussels at locations along the track of the EVOS

through Prince William Sound (PWS) and across the Gulf of Alaska (GOA) (last sampled in 2018 on a five-year cycle).

Over the last 27 years, the program has documented EVOS oil’s disappearance at the spill-impacted sites (albeit

buried oil still exists at a few unique sheltered locations in PWS). Occasionally within the Port, a few tanker- and

diesel-spill incidents have been documented, notably with the recent VMT sump spill in April 2020. Data on mussel

contamination in the VMT spill zone and nearby LTEMP sites, as well as oil-purging rates and gene transcription

responses are being prepared as a separate report and manuscript. Four additional sampling sites were added this

year, two on the Valdez Small Harbor breakwater to assess harbor contamination as possible input to the Port, and

two remote sites in Jack and Galena Bays to validate and/or evaluate alternatives to the Gold Creek (GOC) control

site.

4-12 Attachment A

2020 LTEMP Report

xi

In brief, mussels from the traditional and these added control sites came back exceptionally clean of oil hydrocarbons

except within the immediate vicinity of the VMT spill. For a fifth year, passive sampling devices (PSD) were again

deployed for a month nearby the traditional sites. These devices were intended to compliment the mussel samples

in accumulating only the most bioavailable, dissolved hydrocarbons. And indeed, they returned exceptionally clean

with only a low-level dissolved-phase signal. Subtidal sediment samples again showed that Alaska North Slope (ANS)

oil continues to accumulate at low concentrations near the terminal’s tanker Ballast Water Treatment Facility (BWTF)

discharge outfall.

The program’s field samples (sediments, mussels, and PSD) are processed and analyzed by certified laboratories,

wherein three groups of petroleum hydrocarbons are examined in the data set: polycyclic aromatic hydrocarbons

(PAH), saturated hydrocarbons (SHC), and oil biomarker steranes and terpanes (S/T). The data are then reviewed

from two perspectives: 1) assessing concentrations of oil contaminants, and 2) characterizing the chemical profiles

as to the likely source and degradation state of the hydrocarbons.

Traditionally looking at just total PAH concentrations (conventionally summing 43 analytes into TPAH), it is obvious

that Port Valdez contaminant-oil from Alyeska’s VMT and tanker operations has been trending downward over the

last two decades in both the mussels and sediments. This trend reflects a combination of 1) reduced BWTF discharge

volumes from historically decreasing ANS crude oil production, 2) the transition to double-hulled tankers with

segregated ballast tanks, and 3) improved BWTF efficiency in removing particulate/oil-phase PAH.

From the second perspective, subtly viewing the data as profile patterns, the presence/absence of individual

analytes and their relative concentrations, enables a forensic assessment to determine the source of the oil

components and perhaps their fate as they diminish and weather away in the environment. For example, rather

than just viewing oil contamination by its proxy sum (TPAH), LTEMP mussel profiles from Saw Island (AMT-B),

D

B – Tissue samples

D – Diffuser outfall

P – Passive samplers

S – Sediment grab

S

Valdez

Gold Creek (GOC)

Saw Island

Jackson Point (JAP)

Valdez Marine Terminal

S

AMT

B/P

B/P B/P

(Jack and Galena Bay off map)

Red & Green

2020 Spill

B B

4-12 Attachment A

2020 LTEMP Report

xii

adjacent to the VMT, have been seen generally shifting over the last several years, away from the terminal’s earlier

oil-dominated patterns and into trace-level background or combustion-derived PAH patterns. This assessment task

can be complex or confounded as there are non-oil sources for some analytes in the profiles (i.e., combustion

products and natural hydrocarbons from marine plankton and terrestrial plant waxes). But using all three analyte

groups, PAH, SHC, and biomarkers can create a high confidence in the task. Interpreting the profiles permits insight

into the fate and transport of the oil, information obscured by the common industry practice of just reporting TPAH,

or worse, a subset of PAH.

In 2018, tissue hydrocarbon concentrations at all 11 LTEMP stations (both inside and outside Port Valdez) were

barely detectible as most individual analytes were below the instruments’ Method Detection Limit (MDL) and TPAH

totals reached lows in the 6-40 ng/g dry weight range. Results were similar in 2019, when the three Port Valdez

stations showed the same trace-level components that were essentially indistinguishable from the laboratory

quality-control blanks. At the Gold Creek reference site (GOC), where there had been a minor diesel spill in summer

2016, only trace-level background and combustion profiles were reported in 2018-19. Against these low background

levels, elevated TPAH values and ANS crude oil fingerprints from the April 2020 spill incident at the terminal were

easily detected at the two terminal stations (AMT and JAP).

Comparing nationwide environments, the most recent (albeit dated) National Oceanic and Atmospheric

Administration (NOAA) West Coast Mussel Watch data (2004-05) and the more recent 2008-10 Alaska Mussel Watch

sites, strongly contrast with 2018-2020 LTEMP mussel-tissue results that show 10 to 1,000 times lower TPAH

concentrations. The 2020 LTEMP sites are exceptionally clean of the PAH oil indicators. But during the April 2020

VMT spill incident, mussels within the spill’s “Hot Zone” exhibited TPAH concentrations approaching 230,000 ng/g.

PSDs are specifically designed to sample only the more bioavailable, dissolved hydrocarbons (versus oil

microdroplets that mussels may ingest) and from their month-long submerged deployment, can theoretically

increase the detection sensitivity. All PSD results from the May-June 2020 deployment, however, continued to show

1

10

100

1000

10000

100000

1000000

Jul-

91

Jun

-93

Jun

-95

Jun

-97

Jun

-99

Jun

-01

Jun

-03

Jun

-05

Jun

-07

Jun

-09

Jun

-11

Jun

-13

Jun

-15

Jun

-17

Jun

-19

Jun

-21

Jun

-23

TPA

H4

3 n

g/g

dry

wt

LTEMP MUSSEL TISSUES

AIB-B AMT-BCOH-B GOC-BJAP-B KNH-BSHB-B SHH-BSLB-B WIB-BZAB-B Misc

325617

Eastern Lion tanker spill diesel spills

terminal spills

3933

71

229K

4-12 Attachment A

2020 LTEMP Report

xiii

only a low-level, but heavily weathered, dissolved naphthalene profiles. There was a three-to-five-fold increase in

the reported concentrations at the Saw Island (traditionally, AMT) and Jackson Point (JAP) sites, respectively. But

the PAH patterns were essentially identical to those observed at the Gold Creek (GOC) reference site, which showed

concentrations in line with those observed earlier in 2018 and 2019. In previous year’s deployments both inside and

outside the Port, the commonality of concentrations and patterns suggested that these signals were unrelated to

the BWTF effluent. This year’s elevated PSD concentrations at JAP and AMT probably reflected dissolved-phase PAH

released in the April 2020 spill incident. But it is almost impossible to identify dissolved-PAH sources without other

lines of evidence. We speculate, without water samples, the possibility that the increased concentrations at JAP and

AMT after the April 2020 spill may have broached toxicity thresholds for sensitive marine organisms and life stages.

PSD-measured dissolved PAH concentrations in water at JAP following the April 2020 VMT spill were at a level that

has been shown to cause cardiotoxicity in oil-exposed early life stages of Pacific herring.

These mussel and PSD results differ greatly from the sediments near the terminal that are still a repository for the

BWTF’s chronic hydrocarbon inputs. Sampling bottom sediments near the outfall, the oil compounds are still

measurable at low levels and, in part, still directly traceable to the BWTF discharge. But the patterns are changing.

AMT-S sediment patterns, although historically dominated by an ANS oil signature from the outfall, began changing

to mostly combustion sources in 2011-2015. More recent 2016-2020 patterns again changed and now reflect a mix

of low-level background, combustion, and weathered BWTF oil components. The biomarker profiles (the analytic

hydrocarbon group that are highly resistant to degradation) still solidly confirm that the major oil contaminant

source is the ANS-patterned, BWTF effluent. The 2020 SHC profiles show higher levels of marine and terrestrial

biological components compared to the higher-molecular-weight oil waxes, which have remained relatively constant

between 2008 and 2020. Together, these patterns suggest variable low-level inputs of PAH from weathered ANS oil

(BWTF discharge), plus combustion products from local vessel traffic, runoff, or aerial deposition, and the ubiquitous

trace background. We speculate that being variable and low level, these patterns were likely underlying the earlier

years’ dominant oil signature and are now just more apparent as the total oil concentrations diminish.

1

10

100

1000

10000

Jul-

90

Jul-

92

Jul-

94

Jul-

96

Jul-

98

Jul-

00

Jul-

02

Jul-

04

Jul-

06

Jul-

08

Jul-

10

Jul-

12

Jul-

14

Jul-

16

Jul-

18

Jul-

20

Jul-

22

me

an T

PA

H (

ng/

g d

ry w

t)

AMT-S

GOC-S

114

30

± SE of means

242

47

Eastern Lion tanker spill

LTEMP Sediments

4-12 Attachment A

2020 LTEMP Report

xiv

At the GOC-S reference site, the situation is different: total concentrations are lower, the patterns are mostly from

background and combustion products and there is no evidence of BWTF-derived PAH accumulation in sediments at

the site.

In companion projects, gene transcription data were assessed for the LTEMP mussels and mussels from two

alternate control sites away from the terminal and harbor and nearer to the mouth of Port Valdez (Jack and Galena

Bays). Additional samples were collected near the April 2020 terminal spill location for both oil chemistry and gene

transcription to document acute oiling levels, their relation to gene transcription, and their eventual return to normal

levels. From the panel of selected genes, several showed positive correlation with degree of oiling (i.e., the genes

were turned on to physiologically mitigate exposure to oiling). The LTEMP transcription results are as a separate

report. A manuscript will combine both chemistry and transcription results from the spill samples.

4-12 Attachment A

2020 LTEMP Report

1

INTRODUCTION The primary goal of the Prince William Sound Regional Citizens’ Advisory Council (PWSRCAC) Long-Term

Environmental Monitoring Program (LTEMP) is to monitor impacts from Alyeska’s Valdez Marine Terminal (VMT)

operations and oil transportation activities on the environment at selected sites within Port Valdez, Prince William

Sound (PWS), and Gulf of Alaska (GOA) for “as long as the oil flows through the pipeline.” The project was begun in

1993 as a mandate to the PWSRCAC’s charter. To streamline this report, additional static sections on the project’s

history have been moved into Appendix 3.

The program consists of field samples from sediments, mussels, and passive sampling devices (PSDs), processed and

analyzed by two laboratories: one for tissues, sediments, and occasional water and oil matrices (Alpha

Analytical/NewFields Environmental Forensics), and another for the PSDs (Oregon State University). The labs report

three groups of petroleum hydrocarbons: polycyclic aromatic hydrocarbons (PAH), saturated hydrocarbons (SHC),

and sterane/terpane oil biomarkers (S/T). For LTEMP, the data are then compiled and assessed from two

perspectives: 1) the concentrations of oil contaminants, and 2) characterizing the whole suite of analytes, the

chemical profiles for weathering and source determinations.

From just the profiles, presence/absence patterns of individual analytes and their relative concentrations can

forensically establish the source, transport, and fate of the oil components as they diminish and weather away in

the environment. The task can be complex as there are non-oil sources for some compounds that may be mixed into

the profiles (e.g., combustion products, and natural plankton and plant waxes). The components are generally

described as being from petrogenic (oil sourced), pyrogenic (combustion sourced) or biogenic (natural biological

sourced) origins, or in some mixed combination. In Port Valdez, the levels of oil hydrocarbons have been decreasing

over the past two decades and they are currently at low levels that reflect the reduced inputs from the BWTF

effluent. Spiking above these low levels, spill events are easily detected.

LTEMP sampling locations have been mostly fixed since the program’s 1993 inception. At Alyeska’s VMT, the Ballast

Water Treatment Facility (BWTF) treats and discharges oil-contaminated ballast water offloaded from tankers prior

to onloading the Alaska North Slope (ANS) oil. Here, two stations serve to assess direct exposures from BWTF

effluent: one adjacent to the offshore discharge diffusers near Berth 4 for sediments (AMT-S) and a second at Saw

Island near Berth 5 for mussels (AMT-B). Jackson Point (JAP) was added in 2016 on the opposite (eastern) side of the

diffuser, near Berth 3. Gold Creek (GOC) is sampled as a reference station for both sediments and mussels, 6

kilometers (km) across the Port (Figure 1). Also in 2016, the program was expanded to deploy PSDs at all three Port

stations to measure just the bioavailable, dissolved-phase hydrocarbons. Beyond Port Valdez, eight additional

permanent stations, comprising the geographic reach of the Exxon Valdez oil spill (EVOS) out to Kodiak, were

sampled for mussels but now only every five years (last sampled in 2018) (Figure 2, Table 1). Multi-seasonal sampling

was more frequent during the program’s early years but now has been reduced to just summer or incident sampling.

Sediment samples were also consistently collected throughout the program at two stations, one near the BWTF’s

underwater discharge diffuser and the other, at the GOC reference site across the Port (Figure 1). These are analyzed

for the same hydrocarbon chemistry components as the tissues, plus particle grain size and total organic carbon

content. Sampling and analytical methods are modelled after the protocols developed by the National Oceanic and

Atmospheric Administration (NOAA) Status and Trends Mussel Watch Program as fully detailed in previous annual

monitoring reports.

4-12 Attachment A

2020 LTEMP Report

2

Figure 1. LTEMP sampling stations in Port Valdez adjacent to the terminal (AMT-B, AMT-S, and JAP) and 6 km NW of

the VMT (GOC). New stations in 2020 include Red and Green sites in the Valdez Harbor entrance plus Jack and Galena

Bay in Valdez Arm. This Google Earth image (June 2019) shows a tanker docked at Berth 5.

D

B – Tissue



S – Sediment grab

S

Valdez

Gold Creek (GOC)

Saw Island

Jackson Point (JAP)


S

AMT

B/P

B/P B/P

Red & Green

2020 Spill

B B

VMT

Jack Bay

Galena Bay

4-12 Attachment A

2020 LTEMP Report

3

Figure 2. Map of the LTEMP sites with station abbreviations. PWS and GOA sites are sampled every five years,

whereas Port Valdez sites are sampled annually.

The ensuing report sections introduce and assess the sampled matrices, sediments, tissues, and PSDs. The written

style is intended for the technical reader to fully assess our processes and interpretations; however, many of the

methods and ancillary results sections have been moved to appendices to streamline the main report results.

In this report, the annual results begin with the BWTF effluent characterization. Re-reported here from previous

years (2016-17), this section shows our only examples of what the degraded ANS oil currently looks like as it is

discharged into the Port (versus the fresh ANS oil from the pipeline). We caution that only two seasonal samples,

highly different, were collected to compare to the current field samples. Using these as references against which to

evaluate field samples requires a discerning eye and some flex in judgment but it is possible to confirm or parse out

the presence of ANS oil from biogenic and pyrogenic inputs in a mixed-source sample. Subtle details regarding the

state of weathering (degradation from the original patterns) usually suggests an interpretive scenario of the sources,

transport, and fate for the contaminated sample.

Reported next are results from the sediments near the BWTF outfall that are still accumulating oil, in contrast to the

sediments across the Port at GOC that are generally free of oil. To discriminate just current conditions, the grab-

sampler sediments are collected from just the intact top layer of recently deposited, unconsolidated, fine sediment

(~0.5-1 cm deep). PAH patterns near the outfall are typically extensively weathered (microbially) but the samples

still contain a fairly intact suite of the more recalcitrant S/T biomarkers. Sediments from the GOC reference site

generally do not contain PAH from the BWTF oil, but occasionally, traces of the S/T biomarkers can be observed.

Next sections in this report are the mussel tissues and PSD results, both of which show that, in most samples, there

are essentially only near-method-detection-limit (MDL) traces of oil. Exceptions to these typically low traces were

from the April 2020 VMT spill, as shown in the spill’s “Hot Zone” and nearby Jackson Point (JAP) mussels. Additional

details on depuration kinetics and genetic transcriptomic responses in these samples are contained in a separate

4-12 Attachment A

2020 LTEMP Report

4


spring, summer, autumn (SSA); or summer only (S). Sediments (not shown) have only been sampled in spring and

summer at AMT-S and GOC-S from 1993-2008, and afterwards only in summer.

LTEMP Station Mussel Samplings

Port Valdez Prince William Sound Gulf of Alaska

AMT-B JAP GOC-B KNH DII SLB ZAI SHB COH AIB WIB SHH

1993 SS SS SS SS SS SS SS SS SS SS

1994 SS SS SA SA SA SA SA SA SA SA





1999 SSA SSA SS SS SS SS SS SS SS SS






2005 SSA SSA SS SS SS SS SS S SS SS SS

2006 SSA SSA SS SS SS SS SS SS SS SS SS


2008 SA SSA S S S S S S S S

2009 SS SS S S S S S S S S

2010 S S S

2011 S S S

2012 S S S

2013 S S S S S S S S S S

2014

2015 S S S S

2016 S S S

2017 S S S

2018 S S S S S S S S S S S

2019 S S S

2020 S S S

report (in preparation). Traditionally, the mussels would ingest both dissolved-phase and particulate-phase (micro-

droplets) of oil while the PSDs are designed to sample only dissolved-phase hydrocarbons and, after a month-long

deployment, with more sensitive detection limits. Both sample types agree, showing only background dissolved-

phase contaminants and combustion products in the water column. Furthermore, the low-level PAH concentrations

are too low to be toxic.

4-12 Attachment A

2020 LTEMP Report

5

The mostly static and more ancillary sections from prior reports (e.g., Collection and Analytic Methods, Lab

Performance, Grain Size and TOC, Oxygenated Compounds, and “Beyond LTEMP” that compares LTEMP TPAH with

historic studies) have been moved to appendices. Relevant conclusions still appear in summary points.

Valdez Marine Terminal BWTF Effluent

The primary source of oil contamination in Port Valdez has historically been the partially degraded, ANS crude oil

discharged from the VMT’s BWTF. Last analyzed in 2004-2005 prior to current low production levels and the BWTF

redesign (Payne et al., 2005b, 2005c), re-sampling the effluent was added as an element to the 2016/2017 program.

Sampled during July 2016, the effluent turned out to be nearly 80% freshwater, an unexpectedly low salinity value

that reflects the collected runoff from the terminal and smaller treated tanker-ballast volumes during the summer

months. Anticipating less runoff and more ballast water in the system during winter operations, effluent samples

were again collected in March 2017. For both sampling events, raw effluent as well as filtered samples were obtained

to examine particulate, oil-phase, and dissolved-phase constituents (Payne et al., 1999).

As expected, compared to the summer samples, winter effluent had higher TPAH values (7,605 ng/L vs. 2,885ng/L),

and were less weathered and biodegraded relative to initially fresh ANS crude oil (red line overlay in

Figure 3). During winter, more frequent and stronger winter storms necessitate additional ballast in the tanker cargo

holds and thus, higher volumes and throughput for the BWTF. Also, there is reduced freshwater runoff at the

terminal during the colder winter months. In both seasons, however, particulate/oil-phase droplets were present in

the effluent (Figure 4) at similar concentrations and with essentially identical degrees of weathering. At the same

time, the winter Biological Treatment Tank (BTT) effluent sample had a much higher proportion of bioavailable,

Figure 3. PAH and biomarker profiles (ng/L) of raw (unfiltered) BWTF-BTT effluent samples from July 2016 (upper

plots) and March 2017 (lower plots). The dotted red line represents an overlay of fresh ANS crude oil normalized

(scaled) to hopane (T19, colored gold in the biomarker profiles). Excess dissolved PAH constituents are observed in

2017 as N-, F- and DBT-group analytes above the source reference line (lower left).

0

500

1000

1500

2000

2500

3000

3500

4000

N N1

N2

N3

N4 BI

AC

YA

CN F F1 F2 F3 F4 A Ph

PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L

BWT raw effluent1607003-04

2885Water TPAH7/9/2016

0

100

200

300

400

500

600

T4 T5 T6TR

25b T6

aT6

bT6

cT7 T8 T9 T1

0T1

1T1

1aT1

1b T12

T14a

T14b T1

5T1

6 XT1

7T1

8T1

9N

or3

0H T20

T21

T22

T22A T2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5S2

2 S4 S5 S8 S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS1

TAS2

TAS3

TAS4

ng

/L

BWT raw effluent

7/9/2016

0

200

400

600

800

1000

1200

1400

1600

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L


7605Water TPAH3/20/2017

0

20

40

60

80

100

120

140

T4 T5 T6TR

25b T6

aT6

bT6

cT7 T8 T9 T1

0T1

1T1

1aT1

1b T12

T14a

T14b T1

5T1

6 XT1

7T1

8T1

9N

or3

0H T20

T21

T22

T22A T2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5S2

2 S4 S5 S8 S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS1

TAS2

TAS3

TAS4

ng

/L

BWT raw effluent

3/20/2017

Summer 2016

Winter 2017

ANS reference oil scaled to

sample’s hopane

hopane

PAH Biomarkers

4-12 Attachment A

2020 LTEMP Report

6

dissolved-phase components (Figure 5). It must be cautioned that this profile “snapshot” of the BWTF winter

operation occurred as the BTT was recovering from a shutdown from an overnight power outage; the profile may

be unknowingly biased as normal conditions were reestablished.

Lower-molecular-weight SHC are subject to both dissolution/evaporation losses and microbial degradation (NAS

1975, 1985, and 2003). In a sample’s profile, microbial degradation processes initially appear as decreases in the

more easily assimilated n-alkanes, n-C17 and n-C18, relative to the branched-chain isoprenoids, pristane and phytane.

In the BWTF’s BTT, particulate/oil-phase SHC are well degraded by the combined abiotic and microbial processes in

both seasons, but also partially due to the longer summer residence-time within the tank (Figure 4).

4-12 Attachment A

2020 LTEMP Report

7

Figure 4. PAH, SHC, and biomarker profiles (ng/L) of the filtered, particulate/oil-phase droplets in the BWTF-BTT effluent. The red line overlay represents fresh

ANS scaled to the sample’s hopane (T19, colored gold in the biomarker plots). The gaps between the measured PAH and the overlay portray the extent of

weathering. The TPAH concentrations are similar (1,639 and 2,083 ng/L) but there is additional loss of the higher-molecular-weight (FPs, NBTs, and Cs) in the

summer due to enhanced biodegradation and longer residence time in the BTT. Biomarkers show essentially no degradation in both seasons. SHC (middle plots)

show losses of lower-molecular-weight C9 through C15 components in fresh ANS crude oil largely due to evaporation. In summer SHC, C32 was a matrix interferent;

SHC are scaled to C27.

0

50

100

150

200

250

300

350

400

450

500

T4 T5 T6TR

25b T6

aT6

bT6

cT7 T8 T9 T1

0T1

1T1

1aT1

1b T12

T14a

T14b T1

5T1

6 XT1

7T1

8T1

9N

or3

0H T20

T21

T22

T22A T2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5S2

2 S4 S5 S8 S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS1

TAS2

TAS3

TAS4

ng

/L

BWT effluent filter

7/9/2016

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

1650

C17 Pri

sC

18P

hyt

C19

C20

C21

C22

C23

C24

C25

C26

C27

C28

C29

C30

C31

C32

C33

C34

C35

C36

C37

C38

C39

C40

ng

/L

22,581 TSHCBWT effluent filter

0

500

1000

1500

2000

2500

3000

3500

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L

BWT effluent filter1607004-01

1659Filter (liquid) TPAH7/9/2016 282 Hopane

0

50

100

150

200

250

T4 T5 T6TR

25b T6

aT6

bT6

cT7 T8 T9 T1

0T1

1T1

1aT1

1b T12

T14a

T14b T1

5T1

6 XT1

7T1

8T1

9N

or3

0H T20

T21

T22

T22A T2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5S2

2 S4 S5 S8 S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS1

TAS2

TAS3

TAS4

ng

/L

BWT effluent filter

3/20/2017

0

200

400

600

800

1000

1200

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

1650

C17 Pri

sC

18P

hyt

C19

C20

C21

C22

C23

C24

C25

C26

C27

C28

C29

C30

C31

C32

C33

C34

C35

C36

C37

C38

C39

C40

ng

/L

8,900 TSHCBWT effluent filter

0

200

400

600

800

1000

1200

1400

1600

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L


2083Filter (liquid) TPAH3/20/2017 133 Hopane

PAH Biomarkers SHC

Summer 2016

Winter 2017

4-12 Attachment A

2020 LTEMP Report

8

Figure 5. PAH profiles of BTT effluent samples collected under summer (July 2016) and late winter (March 2017)

conditions: A) whole unfiltered sample; B) particulate/oil phase trapped on the glass-fiber-filter; and C) dissolved

phase. The dotted red lines represent fresh ANS crude oil PAH profile normalized to hopane to show evaporation

and dissolution effects on lower-molecular-weight PAH (C profiles lack the indissolvable hopane for scaling). Note,

however, that while the dissolved-phase patterns are similar, the winter TPAH concentrations are an order of

magnitude higher.

The lower summer BTT throughput requires facility operators to retain and recycle a portion of the BTT contents just

to keep the biological system active. This prolonged recycling practice produces a highly weathered oil profile as

reflected in the two SHC isoprenoids, pristane and phytane, that microbially degrade slower than the straight-chain

alkanes, C17 and C18. Thus, the reduced n-C17/pristane and n-C18/phytane ratios for the oil-phase droplets in the BTT

effluent (Table 2) indicate they have undergone extensive microbial degradation compared to fresh ANS oil.

0

500

1000

1500

2000

2500

3000

3500

4000

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/LBWT raw effluent

1607003-04


0

500

1000

1500

2000

2500

3000

3500

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L


1659Filter (liquid) TPAH7/9/2016

0

200

400

600

800

1000

1200

1400

1600

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L


7605Water TPAH3/20/2017

0

200

400

600

800

1000

1200

1400

1600

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L


2083Filter (liquid) TPAH3/20/2017

0

50

100

150

200

250

300

350

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L

BWT dissolved effluent1607003-05


0

500

1000

1500

2000

2500

3000

3500N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/L

BWT dissolved effluent1703004-04

11295Water TPAH3/20/2017

A

B

C

Summer 2016 Winter 2017

4-12 Attachment A

2020 LTEMP Report

9

Table 2. Ratios of n-C17/pristane and n-C18/phytane for July 2016 and March 2017 BTT effluent samples

(concentrations in ng/L). Lower ratios indicate extent of microbes preferentially degrading the alkanes over the

pristane and phytane isoprenoids.

n-C17 pristane Ratio n-C18 phytane Ratio

ANS Crude Oil 3060 2190 1.40 2710 1410 1.93

July 2016 BTT

Raw effluent 813 944 0.86 602 662 0.91

Particulate phase 658 836 0.79 682 642 1.06

Dissolved phase 243 0 n.a. 0 0 n.a.

March 2017 BTT

Raw effluent 1200 1370 0.88 1140 716 1.59

Particulate phase 359 1040 0.35 548 739 0.74

Dissolved phase 0 290 n.a. 500 189 2.65

PORT VALDEZ SEDIMENTS

In the subsequent discussions, note that we consider TPAH concentrations to be a very rough proxy of oil

contamination (like discussing weather but only talking about the temperature); a truer picture and relevant insights

are in the weathering-profile interpretations. But for historical interests and trend perspectives, TPAH

concentrations are presented and discussed.

SEDIMENT TPAH TRENDS 1993-2020 Between 1993 and 2004, with TPAH levels dropping from historic highs in the hundreds-thousands of ng/g dry weight

(DW) (including a spike in 1995 from the Eastern Lion tanker spill at the terminal), average sediment TPAH

concentrations at the 68-72 meter (m) deep terminal Berth 4 site (AMT-S) continued to decrease from values in the

low hundreds of ng/g in the 2002-2004 period until they dropped to around 50-60 ng/g DW in March 2005. This

decline continued in a range between 20-50 ng/g until April 2012 (Figure 6 and Table 3). Then in 2013, concentrations

unexpectedly dropped further to all-time lows, around 4 ng/g. Sediment samples were not collected in 2014 due to

a temporary hiatus in the program but in July 2015, the TPAH concentrations rebounded slightly to around 15 ng/g

and further up to a range of 55 – 80 ng/g between 2016 and 2019 (Table 3 and Table 4).

The most recent, June 2020 TPAH concentration increase from 66 to 114 ng/g in the AMT-S sediments was at first

believed to possibly suggest a minor contribution from oiled intertidal substrate erosion or oil/suspended

particulate loads and sedimentation after the April 2020 intertidal spill between Berths 4 and 5. The biomarker

profiles observed in the June 2020 AMT-S samples match the April 2020 spilled oil as well as the July 2016 particulate-

phase BWTF effluent reference; however, closer examination of the most recent sediment PAH profiles (discussed

further below) suggests that the increase in the mean TPAH value is actually derived from higher levels of combustion

products in two of the three 2020 AMT sediment replicates.

Sediment TPAH trends at GOC-S have generally tracked with those observed at the terminal (Figure 6), but the

concentrations are usually 2-4 times lower, now around 20-40 ng/g since 2016 (Table 3 and Table 4). The TPAH

concentrations and PAH profiles at GOC-S remained essentially the same in 2019 and 2020.

4-12 Attachment A

2020 LTEMP Report

10

Figure 6. Time series of log (TPAH43) in sediments at AMT-S and GOC-S, 1993-2020.

Table 3. Summary statistics for 2020 TPAH50 and TPAH43 concentrations (ng/g DW).

2020 TPAH50 TPAH43

avg max min count ± SE avg max min count ± SE

AMT-S 130.6 182.1 88.1 3 27.5 113.6 163.0 73.9 3 26.2

GOC-S 30.9 34.4 24.2 3 3.4 29.9 33.0 24.0 3 1.3

Table 4. Historic average sediment TPAH43 values (ng/g DW), 2000-2020.

AMT-S GOC-S AMT-S GOC-S

Apr-00 353 111 Jul-07 65 11

Jul-00 472 374 Jul-08 27 34

Mar-01 828 126 Apr-09 29 12

Jul-01 335 433 Jul-09 17 10

Mar-02 76 312 Jul-10 30 15

Jul-02 464 54 Jul-11 22 8

Mar-03 172 56 Jul-12 25 10

Jul-03 187 32 Jul-13 4 2

Mar-04 167 28 Jul-15 15 3

Jul-04 175 24 Jul-16 56 23

Mar-05 51 19 Jul-17 75 19

Jul-05 86 28 Jun-18 81 20

Mar-06 54 21 Jun-19 66 44

Jul-06 61 16 Jun-20 114 30

Mar-07 35 11

1

10

100

1000

10000

Jul-

90

Jul-

92

Jul-

94

Jul-

96

Jul-

98

Jul-

00

Jul-

02

Jul-

04

Jul-

06

Jul-

08

Jul-

10

Jul-

12

Jul-

14

Jul-

16

Jul-

18

Jul-

20

Jul-

22

me

an T

PA

H (

ng/

g d

ry w

t)

AMT-S

GOC-S

114

30

± SE of means

242

47


LTEMP Sediments

4-12 Attachment A

2020 LTEMP Report

11

SEDIMENT BIOMARKERS Biomarkers’ persistence in the BWTF effluent (and spilled oil) enables tracking the ANS signal in the surrounding

sediments even as the PAH and SHC components are severely degraded. Conceptually, as small oil droplets are

discharged with the effluent, they readily adsorb onto waterborne suspended particulates (e.g., glacial flour) and

eventually settle to the seafloor where microbial degradation and dissolution preferentially weather the SHC and

PAH. In an example profile where PAH are plotted with a fresh ANS profile overlay re-scaled to the sample’s hopane

(Figure 7, top), the individual components almost completely disappear relative to the reference, thus suggesting

nearly complete loss of PAH due to in situ weathering during or after sedimentation (upper-left plot in Figure 7).

Note that there is some degradation/loss of the biomarkers (the small gaps between the hopane-normalized ANS

profile and the individual components in the upper-right panel). Although the biomarkers are recalcitrant (here

microbially non-preferred), they are not invincible. Yet, a sufficiently diagnostic profile exists to confidently assign

the signal to the BWTF effluent. For illustration, when the PAH and biomarker data are re-scaled to the sample’s

NBT2 rather than hopane (Figure 7, bottom plots), another forensic phenomena emerges; an excess of biomarkers

above the fresh-ANS overlay demonstrates the accumulation of slowly degrading biomarkers and slightly less

recalcitrant, higher-alkylated NBT and chrysene homologues over time.

Figure 7. PAH and biomarker profiles of a representative 2016 AMT-S sediment sample overlaid with fresh ANS crude

oil reference (dotted red lines) when alternatively scaled by the highly conservative biomarker, hopane (upper plots),

versus the less recalcitrant PAH, NBT2 (lower plots). The lower plot biomarkers exceeding the reference profile (here

pointing out hopane, colored gold) demonstrate an accumulated excess relative to the more easily degraded,

residual PAH in this sample, plus an accumulation of the more recalcitrant NBT and chrysene PAH homologues.

VALDEZ MARINE TERMINAL SEDIMENTS The PAH, SHC and biomarker profiles from all three 2020 AMT-S sediment samples superficially appear very similar

(Figure 8) but closer examination of PAH patterns from the first two replicates in the upper and middle plots show

significantly elevated levels of combustion products outside of the BWTF particulate-phase template. Parent PAH-

0

20

40

60

80

100

120

140

160

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/g

AMT-S-16-2-2

1607005-02

50Sediment TPAH7/6/2016

0

5

10

15

20

25

30

T4 T5 T6TR

25b T6

aT6

bT6

cT7 T8 T9 T1

0T1

1T1

1aT1

1b T12

T14a

T14b T1

5T1

6 XT1

7T1

8T1

9N

or3

0H T20

T21

T22

T22A T2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5S2

2 S4 S5 S8 S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS1

TAS2

TAS3

TAS4

ng

/g

AMT-S-16-2-2

7/6/2016

0

5

10

15

20

25

N N1

N2

N3

N4 BI

AC

YA


PA

1P

A2

PA

3P

A4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C C1

C2

C3

C4

BB

FB

KF

BA

FB

EPB

AP

PER

IND

PD

AH

AB

GH

I

ng

/g

AMT-S-16-2-2

1607005-02

50Sediment TPAH7/6/2016

0

2

4

6

8

10

12

14

16

18

20

T4 T5 T6TR

25b T6

aT6

bT6

cT7 T8 T9 T1

0T1

1T1

1aT1

1b T12

T14a

T14b T1

5T1

6 XT1

7T1

8T1

9N

or3

0H T20

T21

T22

T22A T2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5S2

2 S4 S5 S8 S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS1

TAS2

TAS3

TAS4

ng

/g

AMT-S-16-2-2

7/6/2016

ANS reference scaled

to sample’s NBT2

fresh ANS crude scaled to

sample’s hopane

weathering

depletion

excess

accumulated

biomarkers surplus residual

NBT and C

PAH Biomarkers

4-12 Attachment A

2020 LTEMP Report

12

dominated phenanthrenes, fluoranthene/pyrenes, chrysenes, and HMW combustion products (BBF through BGHIP)

contribute to the overall measured TPAH value, but these constituents are completely missing in the third replicate

(lower left panel in the figure). The excess parent and alkylated naphthalenes (N, N1, N2, and N3) in all three samples

are most likely derived from the Port’s glacial and riverine sediment inputs (Payne et al., 2010a, 2010b, 2015, Payne

and Driskell, 2017b, Saupe et al., 2005). However, the fluorene, dibenzothiophene, naphthobenzothiophene, and

chrysene groups (Fs, DBTs, NBTs and Cs) in the third replicate (bottom left panel) show an ascending water-washed

pattern consistent with a petrogenic source. The SHC profiles show the almost complete loss of the n-alkane

components that would be expected from the particulate BWTF effluent (or recently spilled ANS oil) due to

evaporation/dissolution processes and microbial degradation. The SHC patterns reflect the typical dominant

contribution from terrestrial plant waxes (n-C25, n-C27, n-C29, and n-C33) with only a minor contribution from higher-

molecular-weight C34 – C40 petroleum waxes. Although the biomarkers are a missing a few components, possibly

reflecting background sediment dilution from riverine and glacial flour, the remaining constituents are a close match

to the expected S/T profile template and demonstrate that they are sourced as ANS oil from the BWTF effluent. The

norhopane (T15)/hopane (T19) ratios and the triaromatic steranes (TAS1 through TAS5) are particularly noteworthy

in this regard. Thus, the overall patterns suggest a mixture of petroleum-sourced and combustion-derived PAH plus

background naphthalenes.

Between 2011 and 2020, the changes in the PAH and SHC patterns reflect different sources and concentration-

dependent, weathering behavior (Figure 9). Specifically, in 2011, the higher-molecular-weight PAH were almost

exclusively pyrogenic as recognized by their patterns of a dominant parent PAH with the alkylated homologues

decreasing in a descending stair-step pattern (note red downward sloping arrows in Figure 9, top panel). This pattern

persisted with the phenanthrenes in 2017 through 2019; however, the other PAH over the last three sampling

periods have been characterized by more of a petrogenic pattern where the parent PAH (FL, DBT, PY, C) within each

group is generally less than the C-2 or C-3 homologues yielding the hump patterns denoted by the red “tents” in

2017 and 2019. In 2020, a mixed source pattern was again noted but with differences among the replicates. Two

replicates (top two panels in Figure 8) had mixed sources, as shown in the bottom profile from the time-series (Figure

9) but the third replicate (bottom profile in Figure 8) is almost exclusively a petrogenic water-washed profile (parent

PAH < C1 < C2 < C3, etc.). This is one of the first instances where source signatures are so dramatically different

among sediment replicates.

The SHC profile trends since 2011 reflect more background, terrestrial-plant-wax, biogenic inputs as reflected in the

alternating spiking of the odd- vs. even-numbered-carbon n-alkanes between n-C23 and n-C29 (Figure 9). In the 2011

SHC plots, the alkanes exhibited a mix of biogenic n-alkanes and higher-molecular-weight C32-C36 petroleum waxes

(Figure 9 top right panel). The 2017 and 2018, AMT-S SHC patterns were very similar in reflecting lower relative

contributions of high-molecular-weight petrogenic waxes compared to 2011. In 2019 and 2020, the petrogenic

waxes were much, much lower compared to the terrestrial plant waxes. Because the absolute concentrations of the

petrogenic waxes in 2017 and 2028 are relatively constant (generally between 40 to 100 ng/g), the increasing

terrestrial-plant-wax contributions since then suggest greater background sediment contributions (riverine and

glacial flour) over this period.

Considering the very low but higher-trending TPAH levels since 2013 (Figure 6), the transitions from pyrogenic

patterns between 2011 and 2015 to petrogenic or mixed petrogenic/combustion sources in 2017 through 2020

presumably reflect a dynamic balance between variable PAH loads from BWTF effluent and accumulation of

pyrogenics (soot). Because background combustion product or soot accumulation at GOC-S is over two-times lower

4-12 Attachment A

2020 LTEMP Report

13

Figure 8. PAH, SHC, and biomarker concentrations and profiles of 2020 AMT-S sediment replicates with mixed background, petrogenic, and combustion sources.

The dotted red line in the PAH and biomarker profiles is the July 2016 particulate-phase BWTF reference (Figure 4 and Figure 5) normalized against the sample’s

hopane; SHC ref normalized to C27. Biomarkers confirm the presence of a weathered ANS profile in the PAH.

0

2

4

6

8

10

12

14

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/gAMT-S-20-2-1

TPAH182

Hopane15

11-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

AMT-S-20-2-1

TSHC3

0

5

10

15

20

25

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

AMT-S-20-2-1

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-20-2-2

TPAH122

Hopane15

11-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

AMT-S-20-2-2

TSHC2

0

5

10

15

20

25

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

AMT-S-20-2-2

0

2

4

6

8

10

12

14

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-20-2-3

TPAH88

Hopane19

11-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

AMT-S-20-2-3

TSHC2

0

5

10

15

20

25

30

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

AMT-S-20-2-3

PAH SHC

S

Biomarkers

4-12 Attachment A

2020 LTEMP Report

14

Figure 9. Representative PAH and SHC signatures of AMT sediments between 2011 and 2020 showing the

progression from a primarily pyrogenic PAH signature to a mix of pyrogenic and water-washed petrogenic

components with increasing terrestrial biogenic SHC and decreasing higher-molecular-weight residual petrogenic

waxes through 2020. Note dominance of 2019 background naphthalenes (N-N4).

(see next section), the likely source at AMT-S is tanker and workboat exhaust while working at or near the berths.

Despite the variable PAH patterns, the biomarker patterns over this timeframe (Figure 10) suggest a consistent,

continued accumulation of BWTF-derived ANS components even as the PAH patterns reflect extensively weathered

oil and the variable addition from combustion sources.

0.00

1.00

2.00

3.00

4.00

5.00

6.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g d

ry

AMT-S-11-2-3

TPAH44

Hopane#N/A

1-Jul-11

0

2

4

6

8

10

12

14

16

18

20

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ng

/g d

ry

AMT-S-11-2-3

TSHC155

0.00

1.00

2.00

3.00

4.00

5.00

6.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-17-2-2

TPAH82

Hopane#N/A

9-Jul-17

0

0.05

0.1

0.15

0.2

0.25

0.3

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

AMT-S-17-2-2

TSHC2

0.00

1.00

2.00

3.00

4.00

5.00

6.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-19-2-3

TPAH94

Hopane#N/A

14-Jun-19

0

200

400

600

800

1000

1200

1400

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

AMT-S-19-2-3

TSHC3,955

0.00

2.00

4.00

6.00

8.00

10.00

12.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-20-2-2

TPAH122

Hopane#N/A

11-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

AMT-S-20-2-2

TSHC2

PAH SHC

2011

2017

2019

2020

petrogenic

waxes in

excess over

other biogenic

alkanes

Parent-PAH

dominated

pyrogenic

patterns

plant wax as

alternating odd

C23-C31 n-alkane

pattern plus HMW

(> C32) petrogenic

waxes.

Water-washed

petrogenic

patterns

pyrogenic and water-washed

petrogenic patterns

Combustion

products

4-12 Attachment A

2020 LTEMP Report

15

Figure 10. Time-series AMT-S sediment PAH profiles from 2011 and 2017 through 2020 along with the biomarker

profiles obtained after introducing those analyses to LTEMP in 2011. The dotted red line denotes the PAH and

biomarker profiles from the July 2016 BWTF particulate-phase normalized to hopane (see Figure 4 top). Biomarkers

confirm ANS oil in all samples. Reporting of TAS biomarkers began in 2017, MAS biomarkers in 2018.

GOLD CREEK SEDIMENTS

Sediments at the shallower (28-30m deep), GOC-S reference site have consistently exhibited lower TPAH

concentrations than AMT-S throughout the duration of the program (Figure 6). In 2013, the GOC-S samples, like

AMT-S, showed record-low PAH concentrations but unlike the sediments at the terminal that rebounded slightly in

2015, the GOC-S levels remained in single digits (no sampling occurred in 2014). TPAH concentrations then increased

modestly from ~6 ng/g in 2015 to around 20 ng/g DW in 2016 and remained at this level through 2018. In 2019, the

TPAH concentrations increased again to around 44 ng/g, which was tentatively attributed to a surge in background

0

1

2

3

4

5

6

7

8

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g d

ry

AMT-S-11-2-3

TPAH44

Hopane11

9-Jun-20

0

2

4

6

8

10

12

14

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-S-11-2-3

0

2

4

6

8

10

12

14

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-17-2-2

TPAH82

Hopane18

9-Jun-20

0

5

10

15

20

25

30

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-S-17-2-2

0

2

4

6

8

10

12

14

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-19-2-3

TPAH94

Hopane20

9-Jun-20

0

5

10

15

20

25

30

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-S-19-2-3

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-20-2-2

TPAH122

Hopane15

9-Jun-20

0

5

10

15

20

25

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-S-20-2-2

PAH Biomarkers

2011

2017

2019

2020

4-12 Attachment A

2020 LTEMP Report

16

naphthalenes and increased levels of combustion products in one replicate. In 2020, the TPAH levels at GOC-S ranged

from 24-33 with a mean of 30 ng/g DW (Figure 6, Table 3). Although the TPAH rise from 6 to 30 ng/g represents a

5-fold increase since 2015, the values are still quite low.

In 2020, remarkable fidelity in both profiles and concentrations was observed in the three GOC sediment grabs

(Figure 11). As in previous years, the PAH profiles suggest little or no petrogenic inputs from the terminal. They are

instead dominated by ubiquitous low-level background naphthalenes and combustion products. This is nicely

demonstrated by the cluster of naphthalenes (N), fluorenes (F), phenanthrenes/anthracenes (PA), and chrysenes (C)

analytes appearing in excess of the red reference overlay of the particulate-phase BWTF effluent (Figure 11). The

SHC patterns show a mix of trace-level marine planktonic alkanes (n-C18 plus odd-carbon-numbered, terrestrial-

plant-wax components (C23, C25, C27, C29, C31) but none of the residual higher-molecular-weight (C33-C40) petrogenic

waxes associated with the terminal (Payne et al., 2015, 2017b).

From 2011 through 2017, the GOC sediment PAH patterns have been essentially identical, dominated by background

naphthalenes and pyrogenics, with TPAH levels modestly increasing (particularly in 2019 due to a spike in higher-

molecular-weight combustion products; Figure 12, bottom panel). The SHC profiles during this period have always

been biogenic, reflecting primarily only background inputs of terrestrial plant waxes. From biomarker data (Figure

13), in addition to natural background biomarkers, low-level traces of some ANS-derived biomarkers are

accumulating in the GOC sediments. While these data are sparse (with many missing components), the observed

biomarkers and diagnostic ratios, norhopane (T15) to hopane (T19), confirm ANS-derived hydrocarbons, presumably

from the BWTF. But despite the biomarkers, there is no evidence of BWTF-derived PAH or SHC accumulation at GOC.

The few biomarkers appear to be the sparse remains of a highly weathered signal transported across the fjord.

The similarity of the PAH and SHC profiles in time-series plots (Figure 12) further supports the notion of a relatively

consistent source over time. In addition to the pyrogenic components, GOC sediments also contain a moderate and

relatively invariant suite of above-MDL parent (N) to N4 naphthalenes (Figure 12), which are also present in the

sediments at AMT-S (Figure 10). These background PAH are believed to derive from glacial and riverine sediment

input to the Port (Payne et al., 2010a, 2010b; Saupe et al., 2005). Similar naphthalene contents were seen to varying

degrees in all 10 major Cook Inlet rivers surveyed during the 2008 EMAP program (ICIEMAP, Susan Saupe, personal

communication, 2009). In these Cook Inlet sediment samples, there was a tentative link to peat inputs (Lees et al.,

2000; Saupe et al., 2005).

4-12 Attachment A

2020 LTEMP Report

17

Figure 11. PAH, SHC, and biomarker profiles of 2020 GOC-S sediment replicates. The dotted red line is July 2016 BWTF particulate phase reference (Figure 4)

normalized against the sample’s hopane; for SHC normalized to C27 (Figure 4). The T32 biomarker spikes are laboratory artifacts.

0

1

1

2

2

3

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/gGOC-S-20-2-1

TPAH24

Hopane2

12-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-S-20-2-1

TSHC2

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

GOC-S-20-2-1

0

1

1

2

2

3

3

4

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-20-2-2

TPAH34

Hopane3

12-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-S-20-2-2

TSHC2

0

1

2

3

4

5

6

7

8

9

10

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

GOC-S-20-2-2

0

1

1

2

2

3

3

4

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-20-2-3

TPAH34

Hopane3

12-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-S-20-2-3

TSHC1

0

1

2

3

4

5

6

7

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

GOC-S-20-2-3

PAH Biomarkers SHC

4-12 Attachment A

2020 LTEMP Report

18

Figure 12. Representative PAH and SHC from GOC-S sediments between 2011 and 2020 showing very similar,

background naphthalene components and pyrogenic, parent-dominated, PAH and higher-molecular-weight

combustion products. SHC patterns and concentrations reflect terrestrial (plant wax) biogenic inputs since 2011. Red

dashed line is sample-specific MDL.

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g d

ry

GOC-S-11-2-2

TPAH8

Hopane1

30-Jun-11

0

5

10

15

20

25

30

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ng

/g d

ry

GOC-S-11-2-2

TSHC71

0.00

0.50

1.00

1.50

2.00

2.50

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-17-2-2

TPAH19

Hopane1

8-Jul-17

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-S-17-2-2

TSHC1

0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

4.00

4.50

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-19-2-3

TPAH59

Hopane3

14-Jun-19

0

100

200

300

400

500

600

700

800C

9C

10C

11C

12C

1313

80C

1414

70C

15C

16N

pri

sC

17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-S-19-2-3

TSHC2,157

0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-20-2-3

TPAH34

Hopane3

12-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-S-20-2-3

TSHC1

pyrogenic

patterns

plant wax as alternating

odd C25-C35 n-alkane

pattern

PAH SHC

2011

2017

2019

2020

background

naphthalenes combustion

products

4-12 Attachment A

2020 LTEMP Report

19

Figure 13. GOC-S sediment PAH and biomarker profiles between 2011 and 2020. The dotted red line is July 2016

BWTF particulate-phase reference. Analyses in 2011 by ABL did not include the triaromatic steroid biomarkers

(TAS); later included in the analyses by Alpha/NewFields.

0

0.2

0.4

0.6

0.8

1

1.2

1.4

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g d

ryGOC-S-11-2-2

TPAH8

Hopane1

9-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-S-11-2-2

0

0.5

1

1.5

2

2.5

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-17-2-2

TPAH19

Hopane1

9-Jun-20

0

2

4

6

8

10

12

14

16

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-S-17-2-2

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-19-2-3

TPAH59

Hopane3

9-Jun-20

0

1

2

3

4

5

6

7

8

9

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-S-19-2-3

0

0.5

1

1.5

2

2.5

3

3.5

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-20-2-3

TPAH34

Hopane3

9-Jun-20

0

1

2

3

4

5

6

7

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-S-20-2-3

pyrogenic

patterns

PAH Biomarkers

2011

2017

2019

2020

4-12 Attachment A

2020 LTEMP Report

20

PORT VALDEZ MUSSEL TISSUES

HISTORICAL TRENDS IN PORT VALDEZ MUSSEL TISSUES Reflecting the changing operations at the terminal, oil discharge into Port Valdez from terminal operations has been

declining over the last two decades. This trend reflects a combination of reduced BWTF discharge volumes from

historically decreased ANS oil production, the transition from single-hulled to double-hulled tankers with segregated

ballast tanks, and improved BWTF efficiency in removing particulate/oil-phase PAH. As a result, over the last several

years, contamination in mussels at the AMT-B sampling site has also been generally shifting away from the terminal’s

earlier petrogenic profiles to background dissolved-phase or pyrogenic (combustion-derived) PAH patterns. As noted

in the sediments section (above), decreasing petrogenic inputs have made the background and pyrogenic inputs

more visible.

Although historically TPAH concentrations in mussels sampled from both the AMT-B and the background-reference

site at GOC-B were commonly reported in hundreds of ng/g, the concentrations dropped to ~80 ng/g levels in 2002

(Figure 14), and with several spill-related exceptions discussed below, they have generally ranged from ~10 to less

than 100 ng/g DW through 2020. The first major exception occurred with a mystery diesel spill at GOC-B in the

summer 2004 when TPAH concentrations approached 1,000 ng/g. By the 2005 collections, the diesel PAH at GOC-B

were long purged and concentrations were back in the pre-spill range around 80 ng/g. They continued to gradually

decline at both locations until 2010, after which the concentrations at GOC-B equaled or slightly exceed those at

AMT-B in 2011 and 2012. In the summer of 2013, collections were very low and only near-MDL, traces of petrogenic

components were present. No samples were collected in 2014 due to a program hiatus, but in 2015, AMT mussels’

PAH increased slightly and transitioned into a primarily dissolved-phase, naphthalene pattern while the GOC-B

tissues were more equivocal (see below). In 2016, another mussel collection site at Jackson Point (JAP), east of the

terminal, was added with the intent of capturing any east-to-west gradients associated with the BWTF discharge. At

that time, the TPAH concentrations near the Terminal ranged from ~70-100 ng/g, while they jumped to 195 ng/g at

GOC-B due to yet another localized diesel spill across the Port. In 2017, the TPAH concentrations at all three Port

Valdez stations ranged from 46 to 63, clustering together around 30-35 ng/g in 2018 and 2019 (Figure 14 and Table

5). At these exceptionally low levels, the individual PAH components at all three Port Valdez sites in 2018-19 were

all below-MDL. Against these low values it was easy to observe the diesel spill at GOC in 2016 and two terminal spills

of ANS crude oil in September 2017 and April 2020 (discussed further below).

Because of the below-MDL concentrations and the presence of the same PAH profiles in all the field samples and

associated laboratory method blanks, source identifications for the routinely collected LTEMP tissue samples from

2018 and 2019 were not possible (Payne and Driskell, 2019, 2020). These findings again reflected the clean

environment from which the samples were collected. However, an isolated and localized September 2017 spill

incident with the Berth 5 Tanker Loading Arm again demonstrated the utility of mussels for monitoring and detecting

oil contamination (Payne and Driskell, 2018b, 2019). Mussels from the event exhibited elevated TPAH levels (Figure

14) from the spill and then, within three months, purged themselves to background levels (Figure 15).

Similarly, in April 2020, an estimated 635 gallons (16 barrels) of ANS crude oil from an overflow sump at the terminal

reached the intertidal zone (ADEC 2020). This resulted in elevated TPAH at all three Port Valdez sites, JAP, AMT and

GOC (438, 256, and 194 ng/g respectively) (Figure 14). The PAH profiles at the traditional LTEMP station at the

eastern Jackson Point site showed a distinct petrogenic signal but by June 2020, the levels had dropped back to the

51-165 ng/g range (discussed further below). Mussels within the “Hot Zone” immediately adjacent to the spill were

heavily oiled (approaching 230,000 ng/g) with obvious ANS-crude-oil profiles. Another report and journal manuscript

are in preparation addressing mussel depuration rates and transcriptomic responses.

4-12 Attachment A

2020 LTEMP Report

21

Figure 14. Time series of mean mussel TPAH43 concentrations comparing 2020 AMT-B, JAP-B and GOC-B with prior

LTEMP collections at other regional sites (open circles). Note the log scale for TPAH concentrations.

Table 5. Time series of mean TPAH43 (ng/g DW, n=3) from AMT-B, GOC-B, and JAP-B mussels, 2008-2020.

Jul-08

Sep-08

Apr-09

Jul-09

Jul-10

Jul-11

Jul-12

Jul-13

Jul-15

Jul-16

Jul-17

Jun-18

Jun-19

Jun-20

mean TPAH43

AMT-B 39 19 38 15 14 30 27 17 51 69 46 35 24 39

GOC-B 58 32 50 21 19 27 45 19 29 195 54 34 70 33

JAP-B 102 63 29 25 71

+ SE of means

AMT-B 5.3 1.8 6.9 0.8 0.9 2.1 2.8 3.0 9.1 13.9 0.8 4.5 12.9 19.9

GOC-B 6.0 3.8 2.8 1.1 1.2 3.0 6.6 2.2 3.1 91.0 3.0 2.2 37.1 16.3

JAP-B 12.0 17.6 0.8 12.7 41.3

1

10

100

1000

10000

100000

1000000

Jul-

91

Jun

-93

Jun

-95

Jun

-97

Jun

-99

Jun

-01

Jun

-03

Jun

-05

Jun

-07

Jun

-09

Jun

-11

Jun

-13

Jun

-15

Jun

-17

Jun

-19

Jun

-21

Jun

-23

TPA

H4

3 n

g/g

dry

wt

AMT-B

JAP-B

GOC-B

other sites

325617


diesel spills

terminal spills

3933

71

229K

4-12 Attachment A

2020 LTEMP Report

22

Figure 15. PAH and biomarker patterns at AMT-B relating to the tanker loading arm spill at the terminal in September

2017. The samples show the background profile from the normal LTEMP collections pre-spill (top, July 2017, TPAH 7

ng/g), the weathered oil in the mussels ~one-week post spill (middle, 108 ng/g), and three months post-spill (bottom,

10 ng/g). Dotted red lines represent fresh ANS crude oil profiles normalized to the sample’s hopane. The below-red-

line gaps (middle left panel) show loss (evaporation and dissolution weathering) of lower-molecular-weight PAH.

Biomarker T26 is a laboratory artifact.

NON-SPILL-RELATED PATTERNS IN PORT VALDEZ MUSSELS

From 2020 LTEMP samplings, representative PAH, SHC, and biomarker plots sampled two months post spill (Figure

16) show near- or below-MDL, mixed pyrogenic-dominated PAH patterns and biogenic SHC. All replicate mussel

samples from the three Port Valdez sites are shown in Appendix 2. There are slightly elevated (and more complex

but still below MDL) traces of residual components from the April 2020 VMT spill at AMT and JAP (seen in water-

washed dibenzothiophene, naphthobenzothiophene, and chrysenes (red tent) PAH patterns) but the GOC profile is

derived solely from local combustion products. The associated quality-control (QC) method blank run with these

samples confirms that the observed PAH in the field samples most likely truly represent trace-level products and not

laboratory artifacts. Identical SHC patterns from these sites reflect only marine biogenic (n-C15, n-C17, and pristane)

plus odd-carbon-number n-alkane traces (n-C23, n-C25, n-C27, n-C29, and n-C31) derived from terrestrial plant

waxes. Biomarker traces were only observed at Jackson Point but with no definitive pattern, no source inference can

be asserted.

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

T4

T5

T6

TR

25

bT6

aT

6b

T6c

T7

T8

T9

T1

0T1

1T1

1aT1

1b T1

2T1

4aT1

4b T1

5T

16 X

T17

T1

8T1

9N

or3

0H

T2

0T

21

T2

2T

22

AT

26

T27

T3

0T3

1T

32

T33

T3

4T

35

S22 S4 S5 S8

S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

Pre

gM

Preg

EPre

gAEP

regB

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

AMT-B-17-2-1

7/8/2017

0

0.5

1

1.5

2

2.5

3

NN

1N

2N

3N

4 BI

AC

YA

CN F

F1 F2 F3 F4 A Ph

PA1

PA2

PA3

PA4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT

1N

BT

2N

BT

3N

BT

4B

AA C

C1

C2

C3

C4

BB

FB

KFB

AF

BE

PB

AP

PER

IND

PD

AH

AB

GH

I

ng

/g

AMT-B-17-2-11707006-01

7Tissue TPAH7/8/2017 0 Hopane

0

5

10

15

20

25

T4

T5

T6

TR

25

bT6

aT

6b

T6c

T7

T8

T9

T1

0T1

1T1

1aT1

1b T1

2T1

4aT1

4b T1

5T

16 X

T17

T1

8T1

9N

or3

0H

T2

0T

21

T2

2T

22

AT

26

T27

T3

0T3

1T

32

T33

T3

4T

35

S22 S4 S5 S8

S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

Pre

gM

Preg

EPre

gAEP

regB

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

SAW-1709-2

9/29/2017

0

20

40

60

80

100

120

140

160

NN

1N

2N

3N

4 BI

AC

YA

CN F

F1 F2 F3 F4 A Ph

PA1

PA2

PA3

PA4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT

1N

BT

2N

BT

3N

BT

4B

AA C

C1

C2

C3

C4

BB

FB

KFB

AF

BE

PB

AP

PER

IND

PD

AH

AB

GH

I

ng

/g

SAW-1709-2L1736093-03

108ET TPAH9/29/2017 14 Hopane

0

2

4

6

8

10

12

14

16

18

T4

T5

T6

TR

25

bT6

aT

6b

T6c

T7

T8

T9

T1

0T1

1T1

1aT1

1b T1

2T1

4aT1

4b T1

5T

16 X

T17

T1

8T1

9N

or3

0H

T2

0T

21

T2

2T

22

AT

26

T27

T3

0T3

1T

32

T33

T3

4T

35

S22 S4 S5 S8

S12

S17

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

Pre

gM

Preg

EPre

gAEP

regB

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

SAW-1712-3

12/7/2017

0

2

4

6

8

10

12

NN

1N

2N

3N

4 BI

AC

YA

CN F

F1 F2 F3 F4 A Ph

PA1

PA2

PA3

PA4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT

1N

BT

2N

BT

3N

BT

4B

AA C

C1

C2

C3

C4

BB

FB

KFB

AF

BE

PB

AP

PER

IND

PD

AH

AB

GH

I

ng

/g

SAW-1712-3L1747478-06


0

5

10

15

20

25

30

35

40

45

50

NN

1N

2N

3N

4 BI

AC

YA

CN F

F1 F2 F3 F4 A Ph

PA1

PA2

PA3

PA4

RET

DB

TD

BT1

DB

T2D

BT3

DB

T4B

BF Fl Py

FP1

FP2

FP3

FP4

NB

TN

BT

1N

BT2

NB

T3N

BT4

BA

A CC

1C

2C

3C

4B

BF

BKF

BA

FB

EP

BA

PPE

RIN

DP

DA

HA

BG

HI

ng

/g

SAW-1709-2L1736093-03


PAH Biomarkers

4-12 Attachment A

2020 LTEMP Report

23

Figure 16. Representative LTEMP Tissue results from June 2020 showing near- or below-MDL, mixed pyrogenic-

dominated PAH patterns and biogenic SHC at AMT, JAP, and GOC along with an associated laboratory method blank.

The dotted red line is the sample’s method detection limit. There is a slightly elevated (but still below MDL) and

more complex PAH pattern at AMT and JAP which may indicate traces of residual PAH from the VMT spill in April

2020 but this is not reflected in either the SHC or biomarker patterns.

VALDEZ MARINE TERMINAL HISTORICAL MUSSEL PATTERNS, PRE-2020 SPILL

To illustrate relevant points in the VMT times series, selected years’ results are presented and discussed in

comparison to the current year. For the AMT site (Figure 17), recent PAH data showed the presence of water-washed

naphthalenes (possibly petrogenic) in 2008 and, in 2015, an increase in dissolved-phase naphthalenes (plus above-

MDL traces of combustion products) (Figure 14). Dissolved-phase naphthalenes were also observed to a lesser extent

in 2015 at GOC (discussed in later section). Recall that the forensically sourcing dissolved-phase PAH is problematic;

as the most abundant and dissolvable PAH, naphthalenes (and other low-molecular-weight PAH) can

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-B-20-2-2

TPAH66

Hopane0

11-Jun-20

0

0.5

1

1.5

2

2.5

3

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

AMT-B-20-2-2

TSHC6

0

0.5

1

1.5

2

2.5

3

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

AMT-B-20-2-2

0

2

4

6

8

10

12

14

16

18

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAP-B-20-2-1

TPAH98

Hopane7

11-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

JAP-B-20-2-1

TSHC5

0

1

2

3

4

5

6

7

8

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

JAP-B-20-2-1

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-B-20-2-1

TPAH59

Hopane0

12-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-B-20-2-1

TSHC4

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

GOC-B-20-2-1

0

0

0

0

0

1

1

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

WG1389790-1-BLANK

TPAH2

Hopane#N/A

0-Jan-00

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

#N/A

WG1389790-1-BLANK

TSHC-

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

#N/A

WG1389790-1-BLANK

AMT-B

JAP-B

GOC-B

Method Blank

PAH SHC Biomarkers

4-12 Attachment A

2020 LTEMP Report

24

derive from any source and cannot be identified from just their profiles. For the 2015 AMT pattern, however, the

naphthalene pattern did not match with BWTF effluent’s naphthalenes (where the parent PAH is depleted in the

processing), so presumably they were derived from another unknown source.

In addition to the dissolved-phase components, parent-PAH-dominated combustion products (e.g., phenanthrene

(Ph), fluoranthenes/pyrenes (FL/PY), and perylene (PER)) are occasionally observed (Figure 17).1 In both 2018 and

2019 (not shown) collections, most of the below-MDL components with the exception of the higher-molecular-

weight (BBF through BGHI) combustion products in 2018 were also associated with the laboratory blanks and were

thus ignored. In June 2020 (two months after the VMT spill), the PAH were mostly at below-MDL levels and showed

a distinct pyrogenic (combustion-derived) source.

The SHC data for 2008 and 2015 AMT samples (Figure 17) show contributions from marine biogenic sources (Payne

et al., 2015) and in the majority of AMT tissues examined since 2008, the SHC have been dominated by biogenic

constituents (e.g., n-C15, n-C17, and pristane) with only very rare observations of petrogenic components. The 2018

and 2020 SHC profiles are nearly identical and show only biogenic-marine-plankton and terrestrial-plant-wax input

with concentrations more that 10-times higher than, and profiles different from, those observed in the method

blanks. Also, there was no indication of the sediment-bound, higher-molecular-weight n-C31 to n-C40 petroleum

waxes (Figure 9) observed in the AMT mussels adjacent to the terminal.

1 Perylene, a 5-ringed PAH, occurs in crude oil but also is naturally generated from biologic processes or early stages of diagenesis

in marine sediments (Bence et al., 2007) and thus, potentially being of non-petroleum origins, is not considered for forensics nor

included in TPAH summations when evaluating non-oil matrices.

4-12 Attachment A

2020 LTEMP Report

25

Figure 17. Time-series mussel PAH and SHC profiles from AMT-B. In 2008, the patterns show primarily below-MDL

water-washed (possibly petrogenic) naphthalenes and combustion products (P/A and FL plus PY). In 2015, there are

above-MDL dissolved-phase naphthalenes and trace-level combustion-product PAHs. In both 2018 and 2019, most

of the below-MDL components are also associated with the lab blank. In June 2020, all of the below MDL PAH are

derived from combustion products. Planktonic biogenic SHC (n-C15, n-C17, and pristane) are also present in most of

the samples. The dotted red line denotes the sample-specific MDL.

0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

4.00

4.50

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g d

ry

AMT-B-08-3-1

TPAH17

Hopane#N/A

29-Sep-08

0

100

200

300

400

500

600

700

800

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ng

/g d

ry

AMT-B-08-3-1

TSHC756

0.00

5.00

10.00

15.00

20.00

25.00

30.00

35.00

40.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g d

ry

AMT-B-15-2-2

TPAH69

Hopane#N/A

2-Jul-15

0

100

200

300

400

500

600

700

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ng

/g d

ry

AMT-B-15-2-2

TSHC1,665

0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

8.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-B-18-2-2

TPAH43

Hopane43

29-Jun-18

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/gAMT-B-18-2-2

TSHC3

0.00

2.00

4.00

6.00

8.00

10.00

12.00

14.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-B-20-2-1

TPAH66

Hopane0

11-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

1.4

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

AMT-B-20-2-1

TSHC5

PAH SHC

2008

2015

2018

2020

4-12 Attachment A

2020 LTEMP Report

26

JACKSON POINT MUSSELS Because mussel samples were only collected from JAP starting in 2016, prolonged time-series data before that are

not available. But comparisons of representative samples from 2016 through June 2020 (Figure 18) show the possible

background contribution of glacial flour/riverine-sourced naphthalenes in 2017 along with trace-level, combustion-

derived PAH in patterns not corresponding with the BWTF effluent. Unfortunately, 2018 and 2019 PAH sources

cannot be assigned because the same components were detected in laboratory method blanks at similar below-MDL

concentrations. The SHC reflect only background marine biogenic components (n-C15, n-C17, and pristane) plus

terrestrial (n-C25, n-C27, and n-C29) plant waxes.

Extra Jackson Point mussels were collected in connection with the April 12, 2020 VMT sump overflow spill. In these

samples, there is unequivocal evidence of the uptake and depuration of PAH and biomarker components from the

spilled oil (Figure 19 and Figure 20). Interestingly, the SHC did not show much accumulation at Jackson Point itself.

Additional mussel collections in the “Hot Zone” closer to the release location did show significant uptake of all three

analyte groups (Figure 19). These findings will be covered in another PWSRCAC report and manuscript (in

preparation).

4-12 Attachment A

2020 LTEMP Report

27

Figure 18. Time-series PAH and SHC profiles of mussels collected at JAP in 2016 through 2020 showing primarily

trace-level dissolved- and (possibly) particulate-phase background naphthalenes plus combustion product PAHs (Ph,

FL, PY, C, BBF, BKE, BEP), and perylene (PER) in 2016 and 2017. The below-MDL PAH in 2018 are suspected of being

procedural artifacts associated with the laboratory method blanks). The dotted red line denotes the sample-specific

MDL.

0.00

2.00

4.00

6.00

8.00

10.00

12.00

14.00

16.00

18.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAP-B-16-2-2

TPAH127

Hopane2

6-Jul-16

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

#N/A

JAP-B-16-2-2

TSHC-

0.00

2.00

4.00

6.00

8.00

10.00

12.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAP-B-17-2-1

TPAH46

Hopane2

8-Jul-17

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

#N/A

JAP-B-17-2-1

TSHC-

0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

8.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAP-B-18-2-3

TPAH29

Hopane0

29-Jun-18

0

0.2

0.4

0.6

0.8

1

1.2C

9C

10C

11C

12C

1313

80C

1414

70C

15C

16N

pri

sC

17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

JAP-B-18-2-3

TSHC4

0.00

2.00

4.00

6.00

8.00

10.00

12.00

14.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAP-B-20-2-3

TPAH73

Hopane5

11-Jun-20

0

0.5

1

1.5

2

2.5

3

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

JAP-B-20-2-3

TSHC6

2016

2017

PAH SHC

2018

2020

4-12 Attachment A

2020 LTEMP Report

28

Figure 19. April 2020 VMT spilled oil, Hot Zone mussels (TPAH 271,000 ng/g DW), and Jackson Point mussels (TPAH 350 ng/g DW) collected approximately three

weeks after the spill. TAS and higher-molecular-weight biomarkers are measured in oil but are not available for mussel extracts (gap on right). The dotted red

line denotes the sample-specific MDL.

PAH SHC Biomarkers

Petrogenic

patterns

Weathered, water-washed

petrogenic PAH Weathered

alkanes plus

biogenics

Sparse but definitive

biomarkers

0

100

200

300

400

500

600

700

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BFL

AB

BF

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

mg/

kgHOT S003

TPAH7940

Hopane289

1-May-20

0

500

1000

1500

2000

2500

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

mg/

kg

HOT S003

TSHC31,298

0

100

200

300

400

500

600

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

mg/

kg

HOT S003

0

5000

10000

15000

20000

25000

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BFL

AB

BF

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ug

/kg

HOT M001

TPAH271306

Hopane3607

30-Apr-20

0

5

10

15

20

25

30

35

40

45

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

mg/

kg

HOT M001

TSHC550

0

500

1000

1500

2000

2500

3000

3500

4000

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ug

/kg

HOT M001

0

5

10

15

20

25

30

35

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BFL

AB

BF

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ug

/kg

JAC M002

TPAH350

Hopane13

1-May-20

0

0.5

1

1.5

2

2.5

3

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

mg/

kg

JAC M002

TSHC6

0

2

4

6

8

10

12

14

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ug

/kg

JAC M002

Spill Oil

April 12, 2020

Hot Zone Mussels

April 30, 2020

Jackson Pt Mussels

May 1, 2020

4-12 Attachment A

2020 LTEMP Report

29

Figure 20. Jackson Point Mussels histograms showing the June 2019 unoiled background pattern, the May 2020 petrogenic profile collected three weeks post

spill, and the June 2020 LTEMP profile demonstrating the recovery to a mixed petrogenic and biogenic pattern. The dotted red line denotes the sample-specific

MDL.

0

1

2

3

4

5

6

7

8

9

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/gJAP-B-19-2-1

TPAH34

Hopane0

15-Jun-19

0

500

1000

1500

2000

2500

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

JAP-B-19-2-1

TSHC4,367

0

0.5

1

1.5

2

2.5

3

3.5

4

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

JAP-B-19-2-1

0

5

10

15

20

25

30

35

40

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ug

/kg

JAC M001

TPAH273

Hopane15

1-May-20

0

0.5

1

1.5

2

2.5

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

mg/

kg

JAC M001

TSHC5

0

2

4

6

8

10

12

14

16

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ug

/kg

JAC M001

0

1

2

3

4

5

6

7

8

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAK-B-20-2-2

TPAH34

Hopane0

9-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

JAK-B-20-2-2

TSHC3

0

1

2

3

4

5

6

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

JAK-B-20-2-2

PAH SHC

June 15, 2019

Biomarkers

May 1, 2020

June 11, 2020

4-12 Attachment A

2020 LTEMP Report

30

GOLD CREEK MUSSELS At GOC, average mussel TPAH levels during the early years of the program (1993-2003) were consistently lower than

or very close to those at AMT-B (Figure 14 and Table 5). In those GOC profiles, mixed dissolved-phase petrogenic

and pyrogenic signals were common and roughly trending with similar-phase patterns or discharge events at AMT-

B (Payne et al., 2008a; 2008b; 2010a; 2015); BWTF oil was commonly present at both sites. After 2002, as TPAH

levels at both stations trended lower, the TPAH levels at GOC have been close to or just slightly above those at AMT-

B, largely due to pyrogenic and occasional petrogenic components--except in 2004 when the PAH and SHC profiles

at GOC documented a fresh diesel spill (Payne et al., 2006). By summer 2005, the diesel signal had largely cleared,

and TPAH levels again generally tracked with AMT through 2015. In 2016, TPAH concentrations at GOC-B increased

dramatically again from 29 to 195 ng/g due to another localized diesel spill (Payne and Driskell, 2019) while the

corresponding levels at AMT-B only increased modestly from 51 to 69 ng/g (Figure 14). By 2017, there was no

evidence of residual diesel at GOC and the TPAH levels at both GOC and AMT-B were 46 and 54 ng/g, respectively.

They have remained in the 40-79 ng/g range through 2020, notwithstanding the ANS crude oil spill at the terminal

in April 2020, and they are generally dominated by combustion products (Figure 16). There was clearly a spike in

TPAH concentrations in the mussel tissues at all three Port Valdez stations in April/May 2020 (Figure 14), but by the

scheduled and routine/traditional LTEMP collections in June, the levels and PAH patterns had almost returned to

background.

The time-series PAH and SHC GOC profiles during non-spill years (2008, 2015, and 2018 through 2020) show variable

sources of dissolved, pyrogenic, and occasionally petrogenic hydrocarbons (Figure 21). In 2008, the at- or just-above-

MDL PAH suggest possible water-washed petrogenic naphthalenes (N-N4), fluorenes (F-F2), and dibenzothiophenes

(DBT-DBT2) plus combustion-derived phenanthrenes (Ph>P/A1>P/A2>P/A3). In 2015, dissolved-phase naphthalene

(N) was the only PAH detected at elevated concentrations (13 ng/g, significantly above MDL) and an even more

complete and descending dissolved-phase (N–N3) naphthalene pattern was also observed at the same time in the

mussels at AMT-B (Figure 17) suggesting a possible common background source. In 2018 and 2019, the GOC PAH

concentrations were all below MDLs (Figure 21) and like the two stations adjacent to the terminal during this period,

the PAH profiles were remarkably similar with only traces of dissolved-phase naphthalenes plus combustion

products at- or just-below MDL. But as noted in previous sections, these patterns were almost identical to those

observed in laboratory method blanks run in parallel with the samples. In 2020, the PAH patterns in the mussels at

GOC were exclusively derived from mostly below MDL combustion products with only the parent PAH from

phenanthrene and fluoranthene above the MDL (Figure 21).

SHC in 2008 and 2015 are mostly trace-level biogenic components (e.g., n-C15, n-C17, and pristane) derived from

marine phytoplankton, algae, and copepods. These same components plus odd-carbon n-alkanes (n-C25, n-C27, n-C29,

n-C31) from terrestrial plant waxes were observed in 2018, 2019, and 2020.

4-12 Attachment A

2020 LTEMP Report

31

Figure 21. Time-series PAH and SHC profiles of GOC mussels collected between 2008 and 2020. The 2008 PAH suggest

possibly particulate-phase, water-washed petrogenic naphthalenes, fluorenes, and DBTs (red tents) plus below-MDL

combustion products. 2015 shows only above-MDL dissolved-phase naphthalenes (also observed at AMT-B at that

time). In 2018 (and 2019, not shown), only at- or below-MDL traces of dissolved naphthalenes and combustion

products are suggested but these same patterns were observed in the laboratory method blanks. The 2020 PAH

profiles are derived exclusively from combustion products. SHC in all years are derived from marine phytoplankton

and copepods. Dotted red line denotes the sample-specific MDLs.

0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g d

ry

GOC-B-08-2-2

TPAH50

Hopane#N/A

21-Jul-08

0

50

100

150

200

250

300

350

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ng

/g d

ry

GOC-B-08-2-2

TSHC651

0.00

5.00

10.00

15.00

20.00

25.00

30.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g d

ry

GOC-B-15-2-2

TPAH25

Hopane#N/A

2-Jul-15

0

50

100

150

200

250

300

350

400

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ng

/g d

ry

GOC-B-15-2-2

TSHC784

0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

8.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-B-18-2-2

TPAH36

Hopane0

29-Jun-18

0

0.1

0.2

0.3

0.4

0.5

0.6

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-B-18-2-2

TSHC4

0.00

2.00

4.00

6.00

8.00

10.00

12.00

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-B-20-2-2

TPAH49

Hopane4

12-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GOC-B-20-2-2

TSHC4

PAH SHC

2008

2015

2018

2020

4-12 Attachment A

2020 LTEMP Report

32

SUPPLEMENTAL MUSSEL STATIONS IN 2020

Four new mussel-sampling stations were added to the 2020-2021 LTEMP collection effort for two purposes: 1) to

resample the Valdez Small Boat Harbor from a matrix more benign than a previous creosote-piling sample (collected

in 2019 as part of the transcriptomics program) and 2) to evaluate other control sites within Port Valdez both for

chemistry and transcriptomics.

Using LTEMP protocols, mussels were collected in the intertidal zone: 1) beneath the red and green harbor lights on

the breakwater riprap entrance to the Valdez Small Boat Harbor and 2) at two more distal locations within the Port,

Jack Bay and Galena Bay (Figure 22). The Jack Bay and Galena Bay stations, further to the west, were added to scope

out potential reference/control sites further from anthropogenic sources associated with the VMT and harbor

activities. They were also intended to support PWSRCAC’s Transcriptomics Project being undertaken by Dr. Lizabeth

Bowen of U.S. Geological Survey (Davis, CA).

Figure 22. Supplemental stations at Galena Bay, Jack Bay, and Valdez Small Boat Harbor entrance (Red and Green

navigation lights). Image from Google Earth dated 12/2016.

The PAH profiles from the Red and Green harbor stations, occupied in June following the April 2020 terminal spill,

show elevated TPAH concentrations (977 and 916 ng/g, respectively; upper plots in Figure 23) with combustion

products dominating the higher-molecular-weight components (parent phenanthrene, fluoranthene, pyrene, and

chrysene with trailing alkylated homologues). The SHC show primarily biogenic n-alkanes and isoprenoids (n-C15, n-

C17, pristane), and higher-molecular-weight, odd-carbon-number, terrestrial plant waxes (n-C23, n-C25, n-C27 and n-

C29). The more complex, underlying pattern of odd and even-carbon numbered n-alkanes in the n-C12 to n-C20 range

plus phytane suggest traces of lighter distillate products (e.g., IFO 180, diesel fuel oil #4; Wang et al., 2007). The PAH

profiles lack the expected patterns for distillates, which either offers little support for the conjecture or they are

overwhelmed by the dominant combustion products. In the biomarker plots, the descending T4-T6 terpanes also

hint at diesel but relative to T4-T6, the other biomarker levels exceed any expectation for diesel and are instead a

close match to the crude ANS oil from the VMT spill. So, it is possible that traces of the spilled oil may have reached

the entrance to the small boat harbor, but the signal is confounded by harbor pyrogenics and diesel contaminants.

Jack Bay

Galena Bay

Red & Green

Valdez

Alyeska Marine

Terminal

Tatitlek

4-12 Attachment A

2020 LTEMP Report

33

PAH profiles from the much cleaner Jack Bay and Galena Bay sites (lower plots in Figure 23) show mostly below-MDL

combustion products at TPAH concentrations more than 30-times lower than the Red and Green harbor sites.

Likewise, from the SHC and biomarker plots, it is apparent that neither Jack Bay nor Galena Bay were contaminated

from the spill oil (there are no tell-tale biomarkers present). The SHC profiles show primarily marine biogenic

components (n-C15, n-C17, pristane) and higher-molecular-weight, odd-carbon-number, terrestrial plant waxes (n-

C23, n-C25, n-C27 and n-C29). At these remote sites, there is no evidence of the evenly repeating series of

n-C12 through n-C20 alkanes plus phytane associated with lighter distillates (Wang et al., 2007).

Figure 23. Representative PAH, SHC, and S/T patterns from the four new stations Red and Green (below the red and

green channel navigation lights on the Valdez Small Boat Harbor entrance breakwater) and Jack Bay and Galena Bay.

The dotted red line represents ANS oil from the VMT spill incident.

GREATER PWS AND GOA STATIONS

Beginning with the 2009 LTEMP program, sampling frequencies in the greater PWS and GOA region were reduced

from twice annually to once every five years. These outer stations were last sampled in 2018 (reported in Payne and

Driskell, 2019). The next sampling will occur in 2023. Like the Port Valdez stations, 2018 TPAH trends continued to

0

50

100

150

200

250

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BFL

AB

BF

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

RED-B-20-2-2

TPAH977

Hopane14

8-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

RED-B-20-2-2

TSHC5

0

2

4

6

8

10

12

14

16

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

RED-B-20-2-2

0

50

100

150

200

250

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BFL

AB

BF

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GRN-B-20-2-2

TPAH916

Hopane25

8-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GRN-B-20-2-2

TSHC5

0

5

10

15

20

25

30

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

GRN-B-20-2-2

0

5

10

15

20

25

30

35

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BFL

AB

BF

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAK-B-20-2-3

TPAH35

Hopane5

9-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

JAK-B-20-2-3

TSHC3

0

1

2

3

4

5

6

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

JAK-B-20-2-3

0

1

2

3

4

5

6

7

8

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BFL

AB

BF

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GAL-B-20-2-1

TPAH27

Hopane0

9-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

C9

C10

C11

C12

C13

1380

C14

1470

C15

C16

Np

ris

C17 Pri

sC

18 Ph

yC

19C

20C

21C

22C

23C

24C

25C

26C

27C

28C

29C

30C

31C

32C

33C

34C

35C

36C

37C

38C

39C

40

ug

/g

GAL-B-20-2-1

TSHC3

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR30

aTR

30b

Tm 14a

14b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA29

SD

IA29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

GAL-B-20-2-1

PAH SHC Biomarkers

Red

Green

Jack Bay

Galena Bay

pyrogenic

patterns

4-12 Attachment A

2020 LTEMP Report

34

decline to all below-MDL lows ranging from 21-38 ng/g (Figure 14) and comprised primarily of components that were

also associated with the laboratory method blanks and occasional variations of the dissolved-phase, background

patterns.

PASSIVE SAMPLING DEVICES

Starting in 2016, the LTEMP program incorporated passive sampling devices (PSDs) deployed in subsurface near-

shore waters adjacent to established LTEMP sites (Figure 24) to monitor PAHs and other petroleum hydrocarbons in

the water column. This sampling effort was motivated by a multi-year trend of observing trace concentrations of

PAHs in mussel tissues, many of which were below the MDLs, and an interest in having data that are toxicologically

relevant to sensitive marine resources, such as early life stages of fish. The goal was to compliment the LTEMP mussel

tissue and sediment data with an integrative, highly sensitive sampling approach that could be used to evaluate the

potential for oil exposure and toxic effects in water-column organisms.

Figure 24. Passive sampling devices (PSD) consisting of a low-density polyethylene membrane enclosed in a stainless-

steel container and deployed subsurface in near-shore subtidal waters adjacent to LTEMP mussel collection sites for

up to 30 days prior to mussel sampling. Sampling photo courtesy of David Janka.

The PSD, a low-density polyethylene membrane in this case, is intended to only sample a fraction of the total

hydrocarbon analytes present, namely, freely dissolved compounds and labile complexes that diffuse into the

membrane that, for biota, are the most bioavailable hydrocarbons. The LTEMP devices were expected to sample

dissolved PAHs and other non-polar or semi-polar hydrocarbons discharged from the BWTF or other sources. The

analytic laboratory at Oregon State University reports 61 PAH isomers as their normal PSD analyte list but in 2018,

the list was expanded to include 40 parent and alkylated PAH homologs used routinely for forensic interpretations.

As a critical part of the method, various deuterated surrogate compounds are pre-infused into the membrane prior

to deployment. Their estimated rate of diffusion out of the membrane while the environmental dissolved-phase

hydrocarbons are infusing calibrates the results for the desired calculation of average dissolved-phase water

concentrations. High detection sensitivity is attained from longer-term deployments in which minute ambient

concentrations are integrated into detectible amounts, similar to how chemicals bioconcentrate in tissues and

organisms.

Beginning in 2016 and 2017, LTEMP PSDs were anchored and constantly submerged for approximately 30 days in

shallow nearshore locations adjacent to the LTEMP mussel sites (Minick and Allan, 2016; Allan, 2018). In 2018, the

program was expanded to encompass Knowles Head (KNH), a clean site outside of Port Valdez located near a

4-12 Attachment A

2020 LTEMP Report

35

transient tanker anchorage, and Disk Island (DII), a site known to contain residual EVOS oil. In 2019 and 2020, only

the three Port Valdez sites were sampled.

In 2020, the PSDs were deployed between May 12 and June 11 at JAP and SAW and June 12 at GOC. PAHs were

detected in the PSDs at all three sites with summed dissolved-PAH concentrations in water (similar to TPAH43) of

213, 68, and 29 ng/L at JAP, SAW, and GOC, respectively. Given their proximity to the April 12, 2020 VMT spill, this

concentration gradient could be expected. A similar trend was also noted in the mussel TPAH values evaluated for

the June spill impacts at these sites (Table 5 and Figure 16). In contrast, the 2019 non-spill-impacted, dissolved-PAH

concentrations were 27.7, 23.7, and 34.9 ng/L at the same three sites.

The dissolved-PAH water concentrations in Port Valdez are low compared to other marine ports in the United States

and comparable to background levels in other parts of Prince William Sound (Lindeberg, Maselko et al. 2017). More

importantly, except for post-spill concentrations at Jackson Point in April 2020, the water concentrations in the Port

are all at least two or three orders of magnitude below published water quality standards and an order of magnitude

below published toxic effects thresholds for aquatic organisms. Again, except for the post-spill concentrations at

JAP, the PSD-derived concentrations of both total PAHs and the sum of 3-ring PAHs in the Port are also less than

demonstrated embryonic exposure concentration thresholds for cardiotoxicity in herring and salmon (Incardona,

Vines et al. 2012, Incardona, Carls et al. 2015). The concentration of dissolved PAHs at JAP following the spill (213

ng/L) is equivalent to summed dissolved-PAH (TPAH43) concentrations shown to cause cardiotoxicity and associated

metabolic impacts in Pacific herring (230 ± 10 ng/L) (Incardona, Carls, et al. 2015). However, the PSD samples from

JAP contained a lower proportion of the three-ring PAHs thought to be primarily associated with cardiotoxicity than

the herring embryos that were exposed to dissolved PAHs from weathered oil in Incardona, Carls, et al 2015.

From a forensic perspective, there is still uncertainty regarding the origin of the PSD signals. The dissolved-phase

PAH patterns were essentially identical at the terminal and the control station (Figure 25) and indeed, for all samples

within and outside the Port throughout the five years’ sampling. All PSD samples have a supra-dominant and largely

invariant evaporatively-weathered naphthalene pattern and two-order-of-magnitude-lower traces of water-washed

alkylated PAH (Figure 25). These observations would strongly suggest some ubiquitous background hydrocarbons

but with the increased and graduated levels correlating with the 2020 spill, the Port Valdez results seem to be truly

recording some form of environmental loadings related to VMT inputs.

The enigma is that the observed dominance of the ascending N<N1<N2<N3<N4 pattern observed at all PSD stations

is the reverse pattern expected from a dissolution process. Specifically, the commonly encountered, dissolved-phase

pattern as based on oil/water partition coefficients (Payne and McNabb, 1984) such that seawater partitioning

against fresh oil or observed in near-shore intertidal waters adjacent to oiled shorelines (Figure 26) shows a more

parent-PAH dominated profile. This pattern was observed from nearshore waters immediately adjacent to

remarkably fresh oil still sequestered in Knight Island intertidal 15 years after EVOS (Payne et al, 2005d). It was also

observed in the dissolved-phase BWTF effluent sampled in March 2017 when the oil was less weathered under

colder, late-winter conditions (Figure 5c, lower right). The reverse pattern with ascending naphthalenes in the PSDs

suggests that to create the pattern either 1) there have been evaporative losses to transform the sequence (slim

possibility to create similar patterns across PWS), 2) some unknown earlier dissolution or aerial process has pre-

determined the pattern that is eventually transported into Port Valdez and PWS waters, or 3) the PSDs are normally

seeing a ubiquitous background but the spill contributed a stronger petrogenic signal on top of the background to

create the gradient.

Still more to consider is that the PSD profiles are similar to a number of the 2013 regional mussel profiles (Payne et

al., 2015). Those low-level, dissolved patterns appearing from Valdez to Kodiak invoked a hypothesis that natural

4-12 Attachment A

2020 LTEMP Report

36

background inputs of PAH were likely derived from large-scale phenomena such as wildfires, glacial melts, riverine

inputs, or terrestrial runoff (e.g., peat and coal are naphthalene rich). As an example of these large-scale processes,

recall that following the EVOS event, there was debate over the unique PAH profile found in the depths of PWS.

Eventually, it was resolved to have originated from source rock formations in the Yakutat region and transported by

coastal currents into PWS (Deepthike et al., 2009). Later Environmental Mapping Project (EMAP) survey work, a joint

project of the Environmental Protection Agency (EPA) and Cook Inlet Regional Citizens Advisory Council, traced those

same offshore, PAH-laden particulates completely across the northern GOA and down the Alaska Peninsula (Saupe

et al., 2005). We suspect that the ubiquitous naphthalene levels in many Port Valdez sediment profiles are tentatively

linked with sediment loads from terrestrial runoff and increased glacial melt. While most of the PAH associated with

these inputs are believed to be very tightly bound to the sediment particles, they may still be a source of low-level

dissolved-phase naphthalenes in the region (Payne et al., 2010b).

With the exceptions of the naphthalene group, concentrations for all other PAH analytes were below 1 ng/L (ppt).

PAH detected at lower levels include fluorenes (F), fluoranthenes (FL), dibenzothiophenes (DBT), phenanthrenes

(Ph), and anthracenes (A) but as dissolved patterns, they defy source characterization. Finally, no matter the source,

from 2016-2020, all the mussels and PSD analyzed throughout all regions (Port Valdez, PWS, and the GOA) were

exceptionally clean with hydrocarbon concentrations below toxicity levels.

4-12 Attachment A

2020 LTEMP Report

37

Figure 25. PAH profiles from 2018 and 2020 PSD deployments. The dominant naphthalenes (blue) are scaled to the

left axis and the two order-of-magnitude lower concentrations of other PAH (Fs. P/As, DBTs, and Cs) (red) are

scaled to the right axis of each plot.

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0

2

4

6

8

10

12

14

16

18

N N1 N2 N3 N4 ACY

ACN F F1 F2 F3 F4

ANT

PHN

PA1

PA2

PA3

PA4

DBT

DBT1

DBT2

DBT3

DBT4 FL P

FP1 C C1 C2 C3 C4 BAA

BBF

BKF

BEP

BAP

PER IP

DAHA

BGHI

P

naph

thale

nes n

g/L

JAC 02-18 TPAH28

0

0.2

0.4

0.6

0.8

1

1.2

0

20

40

60

80

100

120

140

160

N N1 N2 N3 N4 ACY

ACN F F1 F2 F3 F4

ANT

PHN

PA1

PA2

PA3

PA4

DBT

DBT1

DBT2

DBT3

DBT4 FL P


BBF

BKF

BEP

BAP

PER IP

DAHA

BGHI

P

naph

thale

nes n

g/L

JAC-03-20 TPAH205

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0

5

10

15

20

25

N N1 N2 N3 N4 ACY

ACN F F1 F2 F3 F4

ANT

PHN

PA1

PA2

PA3

PA4

DBT

DBT1

DBT2

DBT3

DBT4 FL P


BBF

BKF

BEP

BAP

PER IP

DAHA

BGHI

P

naph

thale

nes n

g/L

SAW 02-18 TPAH35

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

0

10

20

30

40

50

60

N N1 N2 N3 N4 ACY

ACN F F1 F2 F3 F4

ANT

PHN

PA1

PA2

PA3

PA4

DBT

DBT1

DBT2

DBT3

DBT4 FL P


BBF

BKF

BEP

BAP

PER IP

DAHA

BGHI

P

naph

thale

nes n

g/L

SAW-02-20 TPAH88

0

0.2

0.4

0.6

0.8

1

1.2

0

5

10

15

20

25

N N1 N2 N3 N4 ACY

ACN F F1 F2 F3 F4

ANT

PHN

PA1

PA2

PA3

PA4

DBT

DBT1

DBT2

DBT3

DBT4 FL P


BBF

BKF

BEP

BAP

PER IP

DAHA

BGHI

P

naph

thale

nes n

g/L

GOC 01-18 TPAH39

0

0.1

0.2

0.3

0.4

0.5

0.6

0

2

4

6

8

10

12

14

16

18

N N1 N2 N3 N4 ACY

ACN F F1 F2 F3 F4

ANT

PHN

PA1

PA2

PA3

PA4

DBT

DBT1

DBT2

DBT3

DBT4 FL P


BBF

BKF

BEP

BAP

PER IP

DAHA

BGHI

P

naph

thale

nes n

g/L

GOC-01-20 TPAH30

0

0.1

0.2

0.3

0.4

0.5

0.6

0

1

2

3

4

5

6

7

8

9

N N1 N2 N3 N4 ACY

ACN F F1 F2 F3 F4

ANT

PHN

PA1

PA2

PA3

PA4

DBT

DBT1

DBT2

DBT3

DBT4 FL P


BBF

BKF

BEP

BAP

PER IP

DAHA

BGHI

P

naph

thale

nes n

g/L

DSK 02-18 TPAH20

0

0.1

0.2

0.3

0.4

0.5

0.6

0

2

4

6

8

10

12

14

16

18

20

N N1 N2 N3 N4 ACY

ACN F F1 F2 F3 F4

ANT

PHN

PA1

PA2

PA3

PA4

DBT

DBT1

DBT2

DBT3

DBT4 FL P


BBF

BKF

BEP

BAP

PER IP

DAHA

BGHI

P

naph

thale

nes n

g/L

KNH 02-18 TPAH37

PSD Site Abbreviations

JAC – Jackson Point = JAP

SAW – Saw Island = AMT-B

GOC – Gold Creek

DSK – Disk Island = DII

KNH – Knight Island

4-12 Attachment A

2020 LTEMP Report

38

Figure 26. Example PAH profiles of dissolved-phase PAH, A) leaching from remarkably fresh residual Exxon Valdez oil

on Knight Island, 15 years post-spill (from Payne et al., 2005d); B) dissolved-phase BWTF effluent from March 2017.

Descending naphthalene patterns are the compliment to ascending patterns in water-washed particulate oil.

RELATED TOPICS In last year’s report, we presented discussion on topics indirectly related to LTEMP Monitoring. These included: 1)

hydrocarbon bioavailability may not be just limited to dissolved-phase exposures, 2) low level toxicity effects on fish,

3) potential climate change effects on stream flows in relation to enhanced sorption and settling of particulate oil,

and 4) seasonal variability of TPAH due to lipid loss in spawning events. These topics are still relevant but static and

so have been moved to Appendix 9.

A fundamental problem with the LTEMP program is the frequency of sampling. Essentially, the annual data

collections within the Port are equivalent to just a single snapshot of the constantly varying conditions both within

the treatment system, the discharge, stratification of and transport in the receiving waters, subsequent oil

weathering, and the seasonal condition of the mussel populations (feeding, purging, spawning, thermal and

freshwater stresses, etc.). From Alyeska’s Discharge Monitoring Reports (DMR) to EPA (now under ADEC),

documenting the performance and any violations in the BWTF system, various parameters’ time series demonstrate

how little of the variability is captured with LTEMP’s annual, snapshot collections (Figure 27).

0

0.5

1

1.5

2

2.5

3

3.5

4

NN

1N

2N

3N

4 BI

AC

AE F

F1

F2

F3

F4 A P

P/A

1P

/A2

P/A

3P

/A4 D

D1

D2

D3

D4

FL

PY

F/P

1F

/P2

F/P

3F

/P4

BA C

C1

C2

C3

C4

BB

BK

BE

PB

AP

PE

R IP DA

BP

ng/L

TPAH1303704 Dissolved 13KN0117A-waters edge

0

0.5

1

1.5

2

2.5

3

3.5

4

NN

1N

2N

3N

4 BI

AC

AE F

F1

F2

F3

F4 A P

P/A

1P

/A2

P/A

3P

/A4 D

D1

D2

D3

D4

FL

PY

F/P

1F

/P2

F/P

3F

/P4

BA C

C1

C2

C3

C4

BB

BK

BE

PB

AP

PE

R IP DA

BP

TPAH1303704 Dissolv ed 13 ng/LKN0117A-waters edge

0

0.5

1

1.5

2

2.5

3

3.5

4

NN

1N

2N

3N

4 BI

AC

AE F

F1

F2

F3

F4 A P

P/A

1P

/A2

P/A

3P

/A4 D

D1

D2

D3

D4

FL

PY

F/P

1F

/P2

F/P

3F

/P4

BA C

C1

C2

C3

C4

BB

BK

BE

PB

AP

PE

R IP DA

BP

TPAH1303704 Dissolv ed 13 ng/LKN0117A-waters edge

0

500

1000

1500

2000

2500

3000

3500

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/L

BWT dissolved

TPAH11342

Hopane1

20-Mar-17

0

0.5

1

1.5

2

2.5

3

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

µg

/L

BWT dissolved

TSHC9

4-12 Attachment A

2020 LTEMP Report

39

Figure 27. Time series of various DMR parameters as reported in Alyeska’s monthly DMRs from October 2015 through January 2021. Red dots represent LTEMP’s

June or July sampling events to demonstrate the variance not captured by the “snapshot” mussel and sediment collection frequencies.

0

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0.08

0.09

Sep

-15

Dec

-15

Mar

-16

Jun

-16

Sep

-16

Dec

-16

Mar

-17

Jun

-17

Sep

-17

Dec

-17

Mar

-18

Jun

-18

Sep

-18

Dec

-18

Mar

-19

Jun

-19

Sep

-19

Dec

-19

Mar

-20

Jun

-20

Sep

-20

Dec

-20

Mar

-21

Jun

-21

mg/

LAvg and Max BTEX 2015-2020

DMR Avg

DMR Max

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

Sep

-15

Dec

-15

Mar

-16

Jun

-16

Sep

-16

Dec

-16

Mar

-17

Jun

-17

Sep

-17

Dec

-17

Mar

-18

Jun

-18

Sep

-18

Dec

-18

Mar

-19

Jun

-19

Sep

-19

Dec

-19

Mar

-20

Jun

-20

Sep

-20

Dec

-20

Mar

-21

Jun

-21

tho

usa

nd

gal

/day

Avg and Max Flow 2015-2020

Max

Avg

0

10

20

30

40

50

60

Sep

-15

Dec

-15

Mar

-16

Jun

-16

Sep

-16

Dec

-16

Mar

-17

Jun

-17

Sep

-17

Dec

-17

Mar

-18

Jun

-18

Sep

-18

Dec

-18

Mar

-19

Jun

-19

Sep

-19

Dec

-19

Mar

-20

Jun

-20

Sep

-20

Dec

-20

Mar

-21

Jun

-21

mg/

L

Effluent Hydrocarbons 2015-2020

Max Petroleum HC

Avg Aqueous HC

0

2

4

6

8

10

12

Sep

-15

Dec

-15

Mar

-16

Jun

-16

Sep

-16

Dec

-16

Mar

-17

Jun

-17

Sep

-17

Dec

-17

Mar

-18

Jun

-18

Sep

-18

Dec

-18

Mar

-19

Jun

-19

Sep

-19

Dec

-19

Mar

-20

Jun

-20

Sep

-20

Dec

-20

Mar

-21

Jun

-21

mg/

L

Oil and Grease 2015-2020

4-12 Attachment A

2020 LTEMP Report

40

It was fortunate that in 2016 and 2017, our forensics understanding was greatly improved by full analyses of

phase-separated, seasonal effluent samples from the BTT at Alyeska’s BWTF (Figure 3, Figure 4, and Figure

5). These data provided a detailed fingerprint of the dominant hydrocarbon source to the Port, which when

compared to the LTEMP data, enables us to discriminate the petrogenic (oil-based) vs. pyrogenic

(combustion-derived) sources. However, unlike the ANS-source oil, the weathered BWTF effluent has no

standard effluent profile for forensic interpretations. These two seasonally dissimilar profiles emphasize the

effluent’s dynamically changing character. Considering the cycling Oil and Grease DMR chart and its trend-

busting 2020 fluctuations, or similar behavior of the effluent hydrocarbons (Figure 27), a time-series effluent

sampling program might be revealing. If the currently proposed Oxygenated Hydrocarbon project is accepted

by PWSRCAC, this time-series goal may be achieved. But note that additional analyses may be necessary for

full-suite forensic data.

SUMMARY POINTS As oft stated, due to a combination of reduced BWTF discharge volumes from historically decreased North

Slope oil production, the transition to double-hulled tankers with segregated ballast tanks, and improved

BWTF efficiency in recent years, TPAH levels in both sediments and tissues have been trending down since

LTEMP’s inception (see Appendix 3).

In sediments, Port Valdez TPAH levels have been decreasing and reached all-time lows in 2013 but unlike the

trend observed with the mussel tissues (excluding site-specific spill events), sediment TPAH concentrations

at both stations have slightly increased (~50%) over the last four years.

• At the terminal (AMT-S), there has been a transition in the sediment PAH profiles from a dominant

pyrogenic pattern in 2011-2015, to the current mix of background naphthalenes (possibly from

variable riverine and glacial flour input), combustion products, and highly weathered petrogenic

components derived from the BWTF effluent. Biomarker profiles strongly confirm the linkage to the

BWTF effluent throughout the entire period. Recent SHC patterns show a mixture of marine and

terrestrial biogenic components at increasing relative levels compared to the higher-molecular-

weight petrogenic waxes from BWTF discharges that have been observed at this site for years. These

signatures suggest variable or increased background inputs of glacial or riverine flour, weathered

ANS oil from the BWTF, and combustion products from local vessel traffic, runoff, or aerial

deposition.

• At the GOC-S reference site, sediment PAH profiles since 2000 have shown a dominant pyrogenic

pattern with little or no input from the terminal. Instead, they are characterized by variable

naphthalenes and dominant combustion products. SHC profiles continue to be biogenic, reflecting

only phytoplankton sources and terrestrial plant waxes. Trace-level accumulations of biomarkers

associated with the BWTF effluent are accumulating in GOC sediments. They can be attributed to

the terminal even though their associated petrogenic PAH and SHC are largely absent.

• It is speculated that climate-change-accelerated glacial melt and increased stream flows might cause

increased suspended sediment loads and thus, oil adhering to the particles and settling from the

water column (Appendix 9).

TPAH concentrations in mussels at AMT-B, JAP-B, and GOC-B were impacted by the April 2020 VMT spill, but

they largely purged back to normal levels with only slightly elevated levels by the June LTEMP sampling.

• At AMT-B, mussel PAH contamination over the last 10-plus years has been shifting away from the

earlier petrogenic profiles towards trace-level background, dissolved-phase or mixed pyrogenic and

4-12 Attachment A

2020 LTEMP Report

41

petrogenic patterns. In 2018-19 the concentrations were so low, that the PAH patterns could not be

differentiated from trace level components in the laboratory blanks. The immediate impacts of spills

at the terminal were easily detected in mussels collected in September 2017 after the tanker loading-

arm spill but to a lesser extent in May 2020 following the sump overflow incident on April 12, 2020.

• JAP-B mussels have generally tracked those at AMT-B and sampling since 2016 has not demonstrated

any obvious east-west gradients during BWTF operating conditions. Like the 2018-2019 AMT-B

mussels, the concentrations were so low that the PAH patterns could not be differentiated from

laboratory blanks' trace-level components. By way of comparison, the Jackson Point mussels did

show the greatest impacts from the 2020 sump overflow incident (covered in a separate PWSRCAC

report and manuscript in preparation).

• GOC-B mussels have generally shown only low-level pyrogenic PAH profiles since 2006. Like the other

two Port Valdez stations, the 2018 and 2019 PAH concentrations were extremely low with profiles

that could not be differentiated from the laboratory blanks. In the June 2020 collections, the parent

PAH associated with combustion products were again above the MDL, but all other alkylated

homologues were below it. SHC profiles showed only biogenic marine and terrestrial input.

• The mussels from the recently added RED and GRN stations from the riprap entrance to the Valdez

Small Boat Harbor showed significantly elevated (> 900 ng/g) concentrations of combustion products

and possible traces of intermediate fuel oil or diesel from local boat traffic. Additional mussel

sampling along the northern shoreline of the Port between the Harbor and Gold Creek will be needed

to assess the possible influence of combustion products from the Valdez Small Boat Harbor on the

tissues and sediments at Gold Creek.

• The more remote mussel from Jack Bay and Galena Bay showed only trace-level combustion

products and no evidence of any oil from the spill at the VMT.

• Except during the most recent spill incident, current mussel tissue results from the traditional LTEMP

stations are all below TPAH concentrations reported from anywhere else in the United States and

are mostly below even what the National Mussel Watch program categorizes as “low levels” (~63-

1,187 ppb dry weight of PAHs) (Appendix 10).

• These results suggesting an exceptionally clean environment are based on a once-a-year assessment

of tissue (and PSD) contaminant levels. Variable biological and physical conditions plus variable

effluent composition during the rest of the year may greatly influence contaminant levels.

Like their corresponding mussel samples, PSDs showed only a low-level background pattern at all stations;

however, there does appear to be a spatial relationship between measured TPAH concentrations and

proximity to the recent spill at the terminal. With the possible exception of the PSDs from Jackson Point, the

concentrations appear to be below levels of toxic concern.

Pilot analyses of BWTF effluent showed high levels of oxygenated hydrocarbons from weathered ANS oil after

biological degradation in the BTT (Appendix 8). Concentrations of these compounds exceed those of the

analyzed PAH suite. Academic and agency research is ongoing in determining what compounds are present

and which are relevant to toxicity.

In 2016 and 2017, our interpretations were greatly improved by analyses of effluent samples from the BTT at

Alyeska’s BWTF. But these seasonal samples are merely two snapshots of a variable discharge product. We

recommend effluent sampling as a reoccurring component of the program. Also, sampling suspended glacial

flour and river silt from the Valdez Glacier and Lowe Rivers may be warranted to characterize their

hydrocarbon inputs to the Port and further understand the effects of glacial flour scavenging oil in water and

sediment profiles (Appendix 9).

4-12 Attachment A

2020 LTEMP Report

42

CONCLUSIONS For the list of reasons (atop the previous section and in Appendix 3), petrogenic hydrocarbon (oil) inputs into

Port Valdez from the VMT and tanker operations, as reflected in TPAH concentrations in both sediments and

mussels, have been declining.

SEDIMENTS Interpreting the patterns, between 2016 and 2020, the sediments from near the outfall (AMT-S) showed

mixed PAH profiles that included low-level oil components from the BWTF discharge. The profiles were

significantly weathered but were confirmed by the biomarkers to contain ANS-derived oil. The mixture also

suggested variable inputs of background and combustion products (from local vessel traffic, runoff, or aerial

deposition).

At the reference site (GOC-S), the sediments’ PAH patterns are dominated by low-level background

naphthalenes plus combustion products; no oil. The naphthalenes may derive from riverine silt and glacial

flour from nearby Mineral Creek and the Lowe and Valdez Glacier Rivers. Combustion products are likely

introduced by local fishing or pleasure boat traffic, atmospheric input, or as recently hypothesized, possible

contributions from the Valdez Small Boat Harbor. The biomarker data suggests that in addition to natural

background biomarkers, minor residual traces of some ANS-derived biomarkers are accumulating in the GOC

sediments.

TISSUES From 2018-20 mussels, both terminal locations (AMT-B and JAP-B) showed very low-level background PAH

(<70 ng/g) but with no suggestion of BWTF-derived oil.

However, just prior to LTEMP sampling, the April 2020 sump overflow spill at the terminal occurred, which

led to exceptionally high TPAH loads, spiking to ~230,000 ng/g within the spill zone (see Figure 14 and

forthcoming report). Much lower concentrations (438 and 256 ng/g) were observed in May at the nearby

LTEMP stations JAP-B and AMT-B, respectively. When the LTEMP stations were reoccupied in June 2020, the

PAH patterns and concentrations had almost returned to baseline.

GOC-B mussels in both May (spill samples) and June 2020 (LTEMP samples) showed similar low-level

background patterns, including some combustion products; no oil.

PASSIVE SAMPLERS Data from the PSDs corroborate mussel-tissue measurements that only extremely low concentrations of

dissolved-phase PAH are generally present in the region. The 2020 PAH patterns were identical with those

observed in the 2016-2019 series; however, there was a concentration gradient that corresponded with

proximity to the April sump-overflow spill at the terminal. But even at the station closest to the spill (JAP-B),

the observed PSD concentrations from the May to June deployment were below any known toxicity

thresholds for sensitive marine organisms and life stages. The source of the background PAH observed both

within and outside Port Valdez remains enigmatic.

Finally, compared to the recent West Coast Mussel Watch data (2004-05) and the more recent 2008-10 Alaska

Mussel Watch sites, the 2018-20 LTEMP results continue to demonstrate that the sampled region is

exceptionally clean.

4-12 Attachment A

2020 LTEMP Report

43

ACKNOWLEDGEMENTS The beat goes on—but not without the generous support and understanding of our project manager, Austin

Love, the PWSRCAC support staff, and our colleagues on the PWSRCAC Scientific Advisory Committee. Of

course, none of this would happen without the polished field aplomb of our charter skipper, friend, and fellow

scientist, Dave Janka nor the congenial generosity and analytic acumen of our chemistry coordinator, Eric

Litman, of Alpha/NewFields Environmental. And this year, we enthusiastically welcome our new-found

collaborator, Liz Bowen at UC Davis, who’s insights and genetic expertise are taking us in new directions and

adding depth to our interpretations. And again, to our spouses who tolerate our semi-retirement activities.

REFERENCES ADEC 2016. An Evaluation of Remedial Options for Lingering Oil from the Exxon Valdez Oil Spill. Prepared by

the Alaska Department of Environmental Conservation Division of Spill Prevention and Response for

the Exxon Valdez Oil Spill Trustee Council. 18 pgs.

Aeppli, C., C.A. Carmichael, R.K. Nelson, K.L. Lemkau, W.M. Graham, M.C. Redmond, D.L. Valentine, and C.M.

Reddy. 2012. Oil weathering after the Deepwater Horizon disaster led to the formation of

oxygenated residues. Environ. Sci. Technol. 2012, 46 (16), 8799−8807.

Allan, S.E. 2018. Long-Term environmental Monitoring Program – Draft Supplemental report on monitoring

for polycyclic aromatic hydrocarbons using passive samplers: 2016 sampling results and

interpretation. 17 pp.

Bence, A.E., K.A. Kvenvolden, M.C. Kennicutt II. 1996. Organic geochemistry applied to environmental

assessments of Prince William Sound, Alaska, after the Exxon Valdez oil spill—a review. Organic

Geochemistry, Volume 24, Issue 1, January 1996, Pages 7-42, ISSN 0146-6380,

http://dx.doi.org/10.1016/0146-6380(96)00010-1.

Bence, A.E., D.S. Page, and P.D. Boehm. 2007. Advances in forensic techniques for petroleum hydrocarbons:

the Exxon Valdez Experience. Pp 449-487 In Wang, Z. and S.A. Stout (eds). Oil Spill Environmental

Forensics: Fingerprinting and Source Identification. Academic Press, Burlington, MA, USA. 554 pp.

Bouchard, C., M. Geoffroy, M. LeBlanc, A. Majewski, S. Gauthier, W. Walkusz, J.D. Reist, and L Fortier, 2017.

Climate warming enhances polar cod recruitment, at least transiently. Prog. Oceanogr. 156, 121–

129.

Bowen L., A.K. Miles, B. Ballachey, S. Waters, J. Bodkin, M. Lindeberg, D. Esler. 2018. Gene transcription

patterns in response to low level petroleum contaminants in Mytilus trossulus from field sites and

harbors in southcentral Alaska. Deep Sea Research Part II, 147: 27-35.

Bowen L., A. Love, S. Waters, K. Counihan, B. Ballachey, and H. Coletti. 2019. Report: Port Valdez Mussel

Transcriptomics. Prepared for Prince William Sound Regional Citizen’s Advisory Council. Nov 20,

2019. 33 pg.

Brette, F., B. Machado, C. Cros, J.P. Incardona, N.L. Scholz, and B.A Block. 2014. Crude oil impairs cardiac

excitation-contraction coupling in fish. Science 343, 772–776.

Brette, F., H.A. Shiels, G.L.J. Galli, C. Cros, J.P. Incardona, N.L. Scholz, and B.A. Block. 2017. A novel cardiotoxic

mechanism for a pervasive global pollutant. Sci. Rep. 7, 41476.

4-12 Attachment A

2020 LTEMP Report

44

Carls, M.G., S.D. Rice, and J.E. Hose. 1999. Sensitivity of fish embryos to weathered crude oil: Part I. Low-level

exposure during incubation causes malformations, genetic damage, and mortality in larval Pacific

herring (Clupea pallasi). Environ. Toxicol. Chem. 18, 481–493.

Carls, M.G., J.W. Short, and J.R. Payne. 2006. Accumulation of polycyclic aromatic hydrocarbons by

Neocalanus copepods in Port Valdez, Alaska. Marine Pollution Bulletin 52: 1480-1489.

Carls, M.G., Holland L., Pihl E., Zaleski M.A., Moran J., Rice S.D. 2016. Polynuclear Aromatic Hydrocarbons in

Port Valdez Shrimp and Sediment. Arch Environ Contam Toxicol. 71(1):48-59.

Copeman, L.A., B.J. Laurel, M. Spencer, and A. Sremba. 2017. Temperature impacts on lipid allocation among

juvenile gadid species at the Pacific Arctic-Boreal interface: an experimental laboratory approach.

Mar. Ecol. Prog. Ser. 566, 183–198.

Counihan, K.L., L. Bowen, B. Ballachey, H. Coletti, T. Hollmen, B. Pister, T.L. Wilson. 2019. Physiological and

gene transcription assays to assess responses of mussels to environmental changes. PeerJ 7:e7800

https://doi.org/10.7717/peerj.7800

Deepthike, H.U., R. Tecon, G. Van Kooten, J.R. Van der Meer, H. Harms, M. Wells, J. Short. 2009. Unlike PAHs

from Exxon Valdez crude oil, PAHs from Gulf of Alaska coals are not readily bioavailable. Environ Sci

Technol. 2009 Aug 1;43(15):5864-70.

Douglas, G.S. and Liu, B. 2015. Chemical Evidence for Exposure of Red Crabs and Other Deep Benthic

Organisms to Macondo Oil. (CHEM_TR.24). Seattle, WA. DWH Natural Resource Exposure NRDA

Technical Working Group Report.

Driskell, W.B. and J.R. Payne. 2018a. Macondo oil in northern Gulf of Mexico waters – Part 2: Dispersant-

accelerated PAH Dissolution in the Deepwater Horizon plume. Marine Pollution Bulletin 129(1): 412-

419.

Driskell, W. B., and J.R. Payne. 2018b. Development and application of phase-specific methods in oiled-water

forensic studies. Chapter 14 in (S. Stout and Z. Wang, eds). Oil Spill Environmental Forensics Case

Studies. Butterworth-Heinemann/Elsevier Publishers, Oxford, United Kingdom and Cambridge,

Massachusetts. 2018: 289-323.

Edmunds, R.C., J.A. Gill, D.H. Baldwin, T.L. Linbo, B.L. French, T.L. Brown, A.J. Esbaugh, E.M. Mager, J.D.

Stieglitz, R. Hoenig, et al. 2015. Corresponding morphological and molecular indicators of crude oil

toxicity to the developing hearts of mahi mahi. Sci. Rep. 5, 17326.

EPA. 2012. U.S. EPA Fact Sheet for reissue of a National Pollutant Discharge Elimination System (NPDES)

permit to Alyeska Pipeline Service Co. for the Valdez Marine Terminal (AK-002324-8). February 8,

2012. 57 pp.

Hansen, B.H., L. Sørensen, P.A. Carvalho, S. Meier, A.M. Booth, D. Altin, J. Farkas, T. Nordtug. 2018. Adhesion

of mechanically and chemically dispersed crude oil droplets to eggs of Atlantic cod (Gadus morhua)

and haddock (Melanogrammus aeglefinus), Science of The Total Environment, 640–641:138-

143.ISSN 0048-9697, https://doi.org/10.1016/j.scitotenv.2018.05.207.

Helton, D., A. Moles, J. Short, J. Rice. 2004. Results of the M/V Kuroshima Oil Spill Shellfish Tissue Analysis

1999, 2000 and 2004. Report to the M/V Kuroshima Trustee Council. Prepared by NOAA Office of

4-12 Attachment A

https://doi.org/10.7717/peerj.7800

2020 LTEMP Report

45

Response and Restoration, Seattle, Washington and NOAA Auke Bay Laboratory, Juneau, Alaska.

December 2004, 7 pp.

Heintz, R.A. 2007. Chronic exposure to polynuclear aromatic hydrocarbons in natal habitats leads to

decreased equilibrium size, growth, and stability of pink salmon populations. Integr. Environ. Assess.

Manag. 3, 351–363.

Heintz, R.A., S.D. Rice, A.C. Wertheimer, R.F. Bradshaw, F.P. Thrower, F. Joyce, and J.W. Short. 2000. Delayed

effects on growth and marine survival of pink salmon Oncorhynchus gorbuscha after exposure to

crude oil during embryonic development. Mar. Ecol. Prog. Ser. 208, 205–216.

Heintz, R.A., E.C. Siddon, E.V. Farley, and J.M. Napp. 2013. Correlation between recruitment and fall condition

of age-0 pollock (Theragra chalcogramma) from the eastern Bering Sea under varying climate

conditions. Deep Sea Res. II 94, 150–156.

Huckins, J. N.; Petty, J. D.; Booij, K. 2006. Monitors of Organic Chemicals in the Environment: Semipermeable

Membrane Devices; Springer: New York, 2006. 229 pp.

Incardona, J.P. 2017. Molecular mechanisms of crude oil developmental toxicity in fish. Arch. Environ.

Contam. Toxicol. 73, 19–32.

Incardona, J.P., and N.L. Scholz. 2016. The influence of heart developmental anatomy on cardiotoxicity-based

adverse outcome pathways in fish. Aquat. Toxicol. 177, 515–525.

Incardona, J.P., and N.L. Scholz. 2018. Chapter 10: case study: the 2010 Deepwater Horizon oil spill. In

Development, Physiology, and Environment, A. Synthesis, W.W. Burggren, and B. Dubansky, eds.

(Springer), pp. 235–283.

Incardona, J.P., M.G. Carls, L. Holland, T.L. Linbo, D.H. Baldwin, M.S. Myers, K.A. Peck, S.D. Rice, and N.L.

Scholz. 2015. Very low embryonic crude oil exposures cause lasting cardiac defects in salmon and

herring. Sci. Rep. 5, 13499

Incardona, J.P., L.D. Gardner, T.L. Linbo, T.L. Brown, A.J. Esbaugh, E.M. Mager, J.D. Stieglitz, B.L. French, J.S.

Labenia, C.A. Laetz, et al. 2014. Deepwater Horizon crude oil impacts the developing hearts of large

predatory pelagic fish. Proc. Natl. Acad. Sci. U S A 111, E1510–E1518.

Incardona, J.P., C.A. Vines, B.F. Anulacion, D.H. Baldwin, H.L. Day, B.L. French, J.S. Labenia, T.L. Linbo, M.S.

Myers, O.P. Olson, C.A. Sloan, S. Sol, F.J. Griffin, K. Menard, S.G. Morgan, J.E. West, T.K. Collier,

G.M. Ylitalo, G.N. Cherr, N.L. Scholz. 2012. Unexpectedly high mortality in Pacific herring embryos

exposed to the 2007 Cosco Busan oil spill in San Francisco Bay. Proceedings of the National

Academy of Sciences. 109 (2) E51-E58; DOI: 10.1073/pnas.1108884109

Kimbrough, K.L., W. E. Johnson, G.G. Lauenstein, J.D. Christensen and D.A. Apeti. 2008. An Assessment of Two

Decades of Contaminant Monitoring in the Nation’s Coastal Zone. Silver Spring, MD. NOAA Technical

Memorandum NOS NCCOS 74. 105 pp.

Laurel, B.J., L.A. Copeman, P. Iseri, M.L. Spencer, G. Hutchinson, T. Nordtug, C.E. Donald, S. Meier, S.E. Allan,

D.T. Boyd, G.M. Ylitalo, J.R. Cameron, B.L. French, T.L. Linbo, N.L. Scholz, J.P. Incardona. 2019.

Embryonic Crude Oil Exposure Impairs Growth and Lipid Allocation in a Keystone Arctic Forage Fish.

Science. 2019 Sep 27; 19:1101-1113. doi: 10.1016/j.isci.2019.08.051.

4-12 Attachment A

2020 LTEMP Report

46

Lees D.C., W.B. Driskell, J.R. Payne, M.O. Hayes. 2000. Intertidal Reconnaissance Survey in Middle and Upper

Cook Inlet. Report prepared for Cook Inlet Regional Citizens Advisory Council.

Lindeberg, M.R., J. Maselko, R.A. Heintz, C.J. Fugate, L. Holland. 2018. Conditions of persistent oil on beaches

in Prince William Sound 26 years after the Exxon Valdez spill. Deep-Sea Research Part II, 1(47) 9-19.

http://dx.doi.org/10.1016/j.dsr2.2017.07.011

McKenna, A.M., R.K. Nelson, C.M. Reddy, J.J. Savory, N.K. Kaiser, J.E. Fitzsimmons, A.G. Marshall, and R.P.

Rodgers 2013. Expansion of the analytical window for oil spill characterization by ultrahigh resolution

mass spectrometry: beyond gas chromatography. Environ. Sci. Technol. 2013 47 (13), 7530-7539.

DOI: 10.1021/es305284t

Minick, J. and S. Allan. 2016. Long-Term Environmental Monitoring Program – Supplemental Report on monitoring for polycyclic aromatic hydrocarbons using passive samplers: 2016 Sampling Results and Interpretation. PowerPoint presented to PWSRCAC Scientific Advisory Committee.

Morris, J.M., M. Gielazyn, M.O. Krasnec, R. Takeshita, H.P. Forth, J.S. Labenia, T.L. Linbo, B.L. French, J.A. Gill,

D.H. Baldwin, et al. 2018. Crude oil cardiotoxicity to red drum embryos is independent of oil

dispersion energy. Chemosphere 213, 205–214.

National Academy of Sciences (NAS). 1975. Petroleum in the Marine Environment. National Academy of

Sciences, Washington, D.C., 107 pp.

National Academy of Sciences (NAS). 1985. Oil in the Sea: Inputs, Fates, and Effects. National Academy Press,

Washington, D.C., 601 pp.

National Academy of Sciences (NAS). 2003. Oil in the Sea III: Inputs, Fates, and Effects. National Academies

Press, Washington, D.C., 265 pp.

O’Connell, S. G.; Haigh, T.; Wilson, G.; Anderson, K. A. 2013. An analytical investigation of 24 oxygenated-

PAHs (OPAHs) using liquid and gas chromatography−mass spectrometry. Anal. Bioanal. Chem. 2013,

1−12.

Payne, J.R. and W.B. Driskell. 2015a. 2010 DWH offshore water column samples—Forensic assessments and

oil exposures. PECI Technical Report to the Deepwater Horizon Oil Spill Trustees in support of the

PDARP (www.doi.gov/deepwaterhorizon/admin record, DWH-AR0039118, 37 pp.).

Payne, J.R. and W.B. Driskell. 2015b. Forensic fingerprinting methods and classification of DWH offshore

water samples. PECI Technical Report to the Deepwater Horizon Oil Spill Trustees in support of the

PDARP. (www.doi.gov/deepwaterhorizon/admin record, DWH-AR0039170, 31 pp.).

Payne, J.R. and W.B. Driskell. 2016. Water column sampling for forensics. In: Standard Handbook Oil Spill

Environmental Forensics – Fingerprinting and Source Identification (2nd Edition), S. Stout and Z. Wang

(eds.) Elsevier/Academic Press, 2016: 983-1014.

Payne, J.R. and Driskell, W.B. 2017a. Water-column measurements and observations from the Deepwater

Horizon oil spill Natural Resource Damage Assessment. Proceedings of the 2017 Oil Spill Conference.

American Petroleum Institute, Washington, DC. Paper No. 2017-167.

Payne, J.R. and W.B. Driskell. 2017b. Long-Term Environmental Monitoring Program – Final Report: 2016

Sampling Results and Interpretations. Final Report Prepared for Prince William Sound Regional

4-12 Attachment A

2020 LTEMP Report

47

Citizens' Advisory Council Contract No. 951.17.01. Prepared by Payne Environmental Consultants,

Inc., Encinitas, CA. August 2017. 69 pp.

Payne, J.R. and W.B. Driskell. 2018a. Macondo oil in northern Gulf of Mexico waters – Part 1: Assessments

and forensic methods for Deepwater Horizon offshore water samples. Marine Pollution Bulletin

129(1): 399-411.

Payne, J.R. and W.B. Driskell. 2018b. September 2017 Berth 5 Oil Spill – Sampling results and interpretations.

Final Report Prepared for Prince William Sound Regional Citizens' Advisory Council Contract No.

951.18.08. Prepared by Payne Environmental Consultants, Inc., Encinitas, CA. April 2018. 14 pp plus

appendices.

Payne, J.R. and W.B. Driskell. 2018c. Long-Term Environmental Monitoring Program – Final Report: 2017



Inc., Encinitas, CA. September 2018. 60 pp.

Payne, J.R. and W.B. Driskell. 2019. Long-Term Environmental Monitoring Program – Final Report: 2018



Inc., Encinitas, CA. August 2019. 100 pp.

Payne, J.R. and W.B. Driskell. 2020. Long-Term Environmental Monitoring Program – Final Report: 2019



Inc., Encinitas, CA. March 2020. 92 pp.

Payne, J.R. and G.D. McNabb, Jr. 1984. Weathering of petroleum in the marine environment, Marine

Technology Society Journal, 18(3): 24-42.

Payne, J.R., W.B. Driskell, and D.C. Lees. 1998. Long Term Environmental Monitoring Program data analysis of

hydrocarbons in intertidal mussels and marine sediments, 1993-1996. Final Report prepared for the

Prince William Sound Regional Citizens Advisory Council, Anchorage, Alaska 99501. (PWSRCAC

Contract No. 611.98.1). March 16, 1998. 97 pp plus appendices.

Payne, J.R., T.J. Reilly, and D.P. French. 1999. Fabrication of a portable large-volume water sampling system

to support oil spill NRDA efforts. Proceedings of the 1999 Oil Spill Conference, American Petroleum

Institute, Washington, D.C., 1179-1184.

Payne, J.R., W.B. Driskell, M.G. Barron, D.C. Lees. 2001. Assessing transport and exposure pathways and

potential petroleum toxicity to marine resources in Port Valdez, Alaska. Final Report Prepared for

Prince William Sound Regional Citizens' Advisory Council Contract No. 956.02.1. Prepared by Payne

Environmental Consultants, Inc., Encinitas, CA. December 21, 2001. 64 pp plus appendices.

Payne, J.R., W.B. Driskell, M.G. Barron, D.C. Lees, L. Ka’aihue, and J.W. Short. 2003a. Assessing transport and

exposure pathways and potential petroleum toxicity to marine resources in Port Valdez, Alaska.

Poster No. PT214 presented at the SETAC 24th Annual Meeting in North America. November 9-13,

2003, Austin, Texas.

4-12 Attachment A

2020 LTEMP Report

48

Payne, J.R., W.B. Driskell, and J.W. Short. 2003b. 2002-2003 LTEMP Monitoring Report. Final Report prepared

for the Prince William Sound Regional Citizens’ Advisory Council, Anchorage, Alaska 99051.

PWSRCAC Contract No. 951.03.1. Prepared by Payne Environmental Consultants, Inc., Encinitas, CA.

Nov. 6, 2003. 107 pp.

Payne, J.R., W.B. Driskell, M.G. Barron, J. A. Kalmar, and D.C. Lees. 2003c. Public comment regarding the Draft

NPDES Permit for BWTF at Alyeska Marine Terminal. Final Report prepared for the Prince William

Sound Regional Citizens’ Advisory Council, Anchorage, Alaska 99051. PWSRCAC Contract No.

551.02.01. Prepared by Payne Environmental Consultants, Inc., Encinitas, CA. June 2, 2003, 21 p.

Payne, J.R., J.R. Clayton, Jr., and B.E. Kirstein. 2003d. Oil/suspended particulate material interactions and

sedimentation. Spill Science & Technology 8(2): 201-221.

Payne, J.R., W.B. Driskell, and J.W. Short. 2005a. 2003-2004 LTEMP Monitoring Report. Final Report prepared

for the Prince William Sound Regional Citizens’ Advisory Council, Anchorage, Alaska 99051.

PWSRCAC Contract No. 951.04.1. Prepared by Payne Environmental Consultants, Inc., Encinitas, CA.

April 18, 2005. 123 pp.

Payne, J.R., W.B. Driskell, J.F. Braddock, J. Bailey. 2005b. Hydrocarbon biodegradation in the Ballast Water

Treatment Facility, Alyeska Marine Terminal. Final Report prepared for the Prince William Sound

Regional Citizens’ Advisory Council, Anchorage, Alaska 99051. PWSRCAC Contract Numbers

558.04.01 and 560.2004.01. Prepared by Payne Environmental Consultants, Inc., Encinitas, CA. May

2, 2005. 48 pp.

Payne, J.R., W.B. Driskell, J.F. Braddock, J. Bailey, J.W. Short, L. Ka’aihue, T.H. Kuckertz. 2005c. From tankers

to tissues – tracking the degradation and fate of oil discharges in Port Valdez, Alaska. Proceedings of

Arctic Marine Oil Spill Conference 2005, Calgary, Alberta, Canada. pp 959-991.

Payne, J.R. and W.B. Driskell, M.R. Lindeberg, W. Fournier, M.L. Larsen, J.W. Short, S.D. Rice, and D. Janka.

2005d. Dissolved- and particulate-phase hydrocarbons in interstitial water from Prince William

Sound beaches containing buried oil thirteen years after the Exxon Valdez oil spill. Proceedings of

the 2005 International Oil Spill Conference, American Petroleum Institute, Washington, D.C., pp 83-

88.

Payne, J.R., W.B. Driskell, J.W. Short, and M.L. Larsen. 2006. 2004-2005 LTEMP Monitoring Report. Final

Report prepared for the Prince William Sound Regional Citizens’ Advisory Council, Anchorage, Alaska

99051. PWSRCAC Contract No. 951.05.1. (Restoration Project No. 040724). Prepared by Payne

Environmental Consultants, Inc., Encinitas, CA. November 2006. 149 pp.

Payne, J.R., W.B. Driskell, J.W. Short, M.L. Larsen. 2008a. Final 2005-2006 LTEMP Oil Monitoring Report, Exxon

Valdez Oil Spill Restoration Project Final Report (Restoration Project 050763), Prince William Sound

Regional Citizen’s Advisory Council, Anchorage, Alaska. 137 pp.

Payne, J.R., W.B. Driskell, J.W. Short and M.L. Larsen. 2008b. Long-term monitoring for oil in the Exxon Valdez

spill region. Marine Pollution Bulletin 56: 2067-2081.

Payne, J.R., W.B. Driskell, J.W. Short, and M. Larsen. 2010a. Long-Term Environmental Monitoring Program:

Final 2006-2008 LTEMP Monitoring Report. Prince William Sound Regional Citizen’s Advisory Council,

Anchorage, Alaska. 198 pp

4-12 Attachment A

2020 LTEMP Report

49

Payne, J.R., W.B. Driskell, and D. Janka 2010b. A possible source for dissolved-phase PAH signals observed in

Mytilus samples throughout Prince William Sound, Alaska. Proceedings of Arctic Marine Oil Spill

Conference 2010. Halifax, Nova Scotia, CA.

Payne, J.R., W.B. Driskell, J. A. Kalmar. 2012. Review of EPA Draft Permit, Fact Sheet, and other documents

for Proposed Reissuance of Valdez Marine Terminal NPDES Wastewater Discharge Permit (AK-

002324-8). Final Report prepared for the Prince William Sound Regional Citizens’ Advisory Council,

Anchorage, Alaska 99051. PWSRCAC Contract No. 551.09.01. Prepared by Payne Environmental

Consultants, Inc., Encinitas, CA. March 6, 2012. 29 pp.

Payne, J.R., W.B. Driskell, M.G. Carls, M. Larsen, and L.G. Holland. 2013. Long-Term Environmental Monitoring

Program: Results and Interpretations from sampling, 2008-2012. Prince William Sound Regional

Citizen’s Advisory Council, Anchorage, Alaska. 89 pp.

Payne, J.R., W.B. Driskell, M.G. Carls, M.L. Larsen, and L.G. Holland. 2015. Long-Term Environmental

Monitoring Program: Results and Interpretations from sampling, 2008-2013. Prince William Sound

Regional Citizen’s Advisory Council, Anchorage, Alaska. 125 pp.

Payne, J.R., W.B. Driskell, M.G. Carls, and L.G. Holland. 2016. Long-Term Environmental Monitoring Program

– Final Report: 2015 Sampling Results and Interpretations. Prince William Sound Regional Citizen’s

Advisory Council, Anchorage, Alaska. 47 pp.

Peters, K., C. Walters, and J. Moldowan. 2004. The Biomarker Guide: Volume 1 Biomarkers in the Environment

and Human History. 2nd edition. Cambridge: Cambridge University Press. doi:10.1017/

CBO978051152486

Prince William Sound Regional Citizens’ Advisory Council (PWSRCAC). 2019. Public comment on Alaska

Pollutant Discharge Elimination System Individual Permit No. AK0023248 for Alyeska Pipeline Service

Company’s Valdez Marine Terminal. April 26, 2019. 38 pp.

Saupe, S.M., J. Gendron, and D. Dasher. 2005. The Condition of Southcentral Alaska Coastal Bays and

Estuaries. A Statistical Summary for the National Coastal Assessment Program. Alaska Department

of Environmental Conservation. 136 pp.

Shaw, D.G., and A.L Blanchard. 2018. Environmental Studies in Port Valdez, Alaska: 2017. Final Report to

Alyeska Pipeline Service Co., Institute of Marine Science, University of Alaska Fairbanks, 127 pp.

Shaw, D.G., and A.L Blanchard. 2019. Environmental Studies in Port Valdez, Alaska: 2018. Final Report to

Alyeska Pipeline Service Co., Institute of Marine Science, University of Alaska Fairbanks, 126 pp.

Short, J.W., G.V. Irvine, D.H. Mann, J.M. Maselko, J.J. Pella, M.R. Lindeberg, J.R. Payne, W.B. Driskell, and S.D.

Rice. 2007. Slightly weathered Exxon Valdez oil persists in Gulf of Alaska beach sediments after 16

years. Environmental Science & Technology. 41(4): 1245-1250.

Sørensen, L., E. Sørhus, T. Nordtug, J.P. Incardona, T.L. Linbo, L. Giovanetti, O. Karlsen, O., and S. Meier. 2017.

Oil droplet fouling and differential toxicokinetics of polycyclic aromatic hydrocarbons in embryos of

Atlantic haddock and cod. PLoS One 12, e0180048.

Sørensen, L., P. McCormack, D. Altin, W.J. Robson, A.M. Booth, L.-G. Faksness, S.J. Rowland, T.R. Størseth.

2019. Establishing a link between composition and toxicity of offshore produced waters using

4-12 Attachment A

2020 LTEMP Report

50

comprehensive analysis techniques – A way forward for discharge monitoring? Science of the Total

Environment 694, 133682.

Sørhus, E., R.B. Edvardsen, Ø. Karlsen, T. Nordtug, T. van der Meeren, A. Thorsen, C. Harman, S. Jentoft, S.

Meier. 2015. Unexpected interaction with dispersed crude oil droplets drives severe toxicity in

Atlantic Haddock embryos. PLoS ONE 10(4): e0124376. https://doi.org/10.1371/journal.

pone.0124376

Sørhus, E., J.P. Incardona, Ø. Karlsen, T.L. Linbo, L. Sørensen, T. Nordtug, T. van der Meeren, A. Thorsen, M.

Thorbjørnsen, S. Jentoft, R.B. Edvardsen and S. Meier. 2016. Effects of crude oil on haddock reveal

roles for intracellular calcium in craniofacial and cardiac development. Sci. Rep. 6, 31058.

Sower, G.J., K.A. Anderson. 2008. Spatial and temporal variation of freely dissolved polycyclic aromatic

hydrocarbons in an urban river undergoing superfund remediation. Environmental Science &

Technology. 2008, 42 (24), 9065−9071.

Stout, S.A. and J.R. Payne. 2016a. Chemical composition of floating and sunken in-situ burn residues from the

Deepwater Horizon oil spill. Marine Pollution Bulletin 108: 186-202.

Stout, S.A. and J.R. Payne. 2016b. Macondo oil in deep-sea sediments: Part 1 – sub-sea weathering of oil

deposited on the seafloor. Marine Pollution Bulletin, 111, 365-380.

Stout, S.A. and J.R. Payne. 2017. Footprint, weathering, and persistence of synthetic-base drilling mud olefins

in deep-sea sediments following the Deepwater Horizon disaster. Marine Pollution Bulletin 118: 328-

340.

Stout, S.A., J.R. Payne, S.D. Emsbo-Mattingly, and G. Baker. 2016a. Weathering of field-collected floating and

stranded Macondo oils during and shortly after the Deepwater Horizon oil spill. Marine Pollution

Bulletin 105: 7-22.

Stout, S.A., J.R. Payne, R.W. Ricker, G. Baker, C. Lewis. 2016b. Macondo oil in deep-sea sediments: Part 2 –

Distribution and distinction from background and natural oil seeps. Marine Pollution Bulletin 111:

381-401.

Stout, S.A. and Z. Wang, 2007. Chemical fingerprinting of spilled or discharged petroleum – methods and

factors affecting petroleum fingerprints in the environment. Chapter 1, Oil Spill Environmental

Forensics – Fingerprinting and Source Identification. 1st Edition. Wang, Z. and S.A. Stout (eds),

Academic Press (an imprint of Elsevier), Burlington, MA. 554 pp. ISBN 10: 0-12-369523-6.

Stout, S.A. and Z. Wang, 2016. Chemical Fingerprinting Methods and Factors Affecting Petroleum Fingerprints

in the Environment. Chapter 3, Standard Handbook Oil Spill Environmental Forensics: Fingerprinting

and Source Identification. 2nd Edition. Stout, S.A. and Z. Wang (eds), Academic Press, Burlington, MA.

1107 pp. DOI: 10.1016/B978-0-12-809659-8.00003-6

Wang, Z, C. Yang, M. Fingas, B. Hollebone, L.H. Yim, and J.R. Oh, 2007. Petroleum biomarker fingerprinting

for oil spill characterization and source identification. Chapter 3, Oil Spill Environmental Forensics –

Fingerprinting and Source Identification. 1st Edition. Wang, Z. and S.A. Stout (eds), Academic Press

(an imprint of Elsevier), Burlington, MA. 554 pp. ISBN 10: 0-12-369523-6.

4-12 Attachment A

2020 LTEMP Report

51

APPENDIX 1. POLYCYCLIC AROMATIC HYDROCARBON (PAH), SATURATED

HYDROCARBON (SHC), AND BIOMARKER ANALYTES

Analytes Abbreviation

Naphthalene N

C1-Naphthalene N1

C2-Naphthalene N2

C3-Naphthalene N3

C4-Naphthalene N4

Biphenyl BI

Acenaphthylene ACY

Acenaphthene ACN

Fluorene F

C1-Fluorene F1

C2-Fluorene F2

C3-Fluorene F3

C4-Fluorene F4

Anthracene A

Phenanthrene Ph

C1-Phenanthrene/Anthracene PA1




Retene RET

Dibenzothiophene DBT

C1-Dibenzothiophene DBT1




Benzo(b)fluorene BF

Analytes Abbreviation

Fluoranthene FL

Pyrene PY

C1-Fluoranthene/Pyrene FP1




Napthobenzothiophene NBT

C1-Napthobenzothiophene NBT1




Benzo(a)Anthracene BAA

Chrysene C

C1-Chrysene C1

C2-Chrysene C2

C3-Chrysene C3

C4-Chrysene C4

Benzo(b)fluoranthene BBF

Benzo(k)fluoranthene BKF

Benzo(e)pyrene BEP

Benzo(a)pyrene BAP

Perylene PER

Indeno(1,2,3-cd)pyrene IND

Dibenzo(a,h)anthracene DAHA

Benzo(g,h,i)perylene BGH

Total PAH TPAH

ANS Crude oil example

0

200

400

600

800

1000

1200

1400

1600

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F

F1 F2 F3A

NT

PH

NP

A1

PA

2P

A3

PA

4R

ETD

BT0

DB

T1D

BT2

DB

T3D

BT4

CA

RB

BB

FFL

AP

YR FP1

FP2

FP3

FP4

NB

TN

BT1

NB

T2N

BT3

NB

T4B

AA C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

mg

/kg

WG1370244-1-ANS

TPAH9395Hopane#N/A

0-Jan-00

4-12 Attachment A

2020 LTEMP Report

52

Saturated hydrocarbons (SHC or n-alkanes)

Analyte Abbrev

Nonane (C9) C9

Decane (C10) C10

Undecane (C11) C11

Dodecane (C12) C12

Tridecane (C13) C13

2,6,10 Trimethyldodecane (1380) 1380

Tetradecane (C14) C14

2,6,10-Trimethyltridecane (1470) 1470

Pentadecane (C15) C15

Hexadecane (C16) C16

Norpristane (1650) Pristane

Heptadecane (C17) C17

Pristane Phytane

Octadecane (C18) C18

Phytane Phy

Nonadecane (C19) C19

Eicosane (C20) C20

Heneicosane (C21) C21

Docosane (C22) C22

Tricosane (C23) C23

Tetracosane (C24) C24

Pentacosane (C25) C25

Hexacosane (C26) C26

Heptacosane (C27) C27

Octacosane (C28) C28

Nonacosane (C29) C29

Triacontane (C30) C30

Hentriacontane (C31) C31

Dotriacontane (C32) C32

Tritriacontane (C33) C33

Tetratriacontane (C34) C34

Pentatriacontane (C35) C35

Hexatriacontane (C36) C36

Heptatriacontane (C37) C37

Octatriacontane (C38) C38

Nonatriacontane (C39) C39

Tetracontane (C40) C40

Total SHC TSHC

0

1000

2000

3000

4000

5000

6000

7000

C9

C1

0C

11

C1

2C

13

13

80

C1

41

47

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

mg

/kg

WG1307766-1-ANS

TSHC65,519

4-12 Attachment A

2020 LTEMP Report

53

Petroleum Biomarkers

Class Biomarker Abbrev

Terpanes C23 Tricyclic Terpane (T4) T4

C24 Tricyclic Terpane (T5) T5

C25 Tricyclic Terpane (T6) T6

C24 Tetracyclic Terpane (T6a) T6a

C26 Tricyclic Terpane-22S (T6b) T6b

C26 Tricyclic Terpane-22R (T6c) T6c

C28 Tricyclic Terpane-22S (T7) T7

C28 Tricyclic Terpane-22R (T8) T8

C29 Tricyclic Terpane-22S (T9) T9

C29 Tricyclic Terpane-22R (T10) T10

18a-22,29,30-Trisnorneohopane-TS (T11)

Ts

C30 Tricyclic Terpane-22S C30Ts

C30 Tricyclic Terpane-22R C30Tr

Hopanes 17a(H)-22,29,30-Trisnorhopane-TM

Tm

17a/b,21b/a 28,30-Bisnorhopane (T14a)

14a

17a(H),21b(H)-25-Norhopane (T14b)

14b

30-Norhopane (T15) T15

18a(H)-30-Norneohopane-C29Ts (T16)

T16

17a(H)-Diahopane (X) X

30-Normoretane (T17) T17

18a(H)&18b(H)-Oleananes (T18) T18

Hopane (T19) T19

Moretane (T20) T20

30-Homohopane-22S (T21) T21

30-Homohopane-22R (T22) T22

Gammacerane/C32-Diahopane T22a

30,31-Bishomohopane-22S (T26) T26

30,31-Bishomohopane-22R (T27) T27

30,31-Trishomohopane-22S (T30) T30

30,31-Trishomohopane-22R (T31) T31

Tetrakishomohopane-22S (T32) T32

Tetrakishomohopane-22R (T33) T33

Pentakishomohopane-22S (T34) T34

Pentakishomohopane-22R (T35) T35

Steranes 13b(H),17a(H)-20S-Diacholestane (S4) S4

13b(H),17a(H)-20R-Diacholestane (S5) S5


13b,17a-20S-Methyldiacholestane (S8) S8

17a(H)20SC27/C29dia DIA29S

17a(H)20rc27/C29dia DIA29R

Unknown Sterane (S18) S18

13a,17b-20S-Ethyldiacholestane (S19) S19

14a,17a-20S-Methylcholestane (S20) S20

14a,17a-20R-Methylcholestane (S24) S24

14a(H),17a(H)-20S-Ethylcholestane (S25) S25

14a(H),17a(H)-20R-Ethylcholestane (S28) S28

14b(H),17b(H)-20R-Cholestane (S14) S14

14b(H),17b(H)-20S-Cholestane (S15) S15

14b,17b-20R-Methylcholestane (S22) S22

14b,17b-20S-Methylcholestane (S23) S23

14b(H),17b(H)-20R-Ethylcholestane (S26) S26

14b(H),17b(H)-20S-Ethylcholestane (S27) S27

C20 Pregnane Preg

C21 20-Methylpregnane MPreg

C22 20-Ethylpregnane (a) EPregA

C22 20-Ethylpregnane (b) EPregB

Triaromatic Steroids C26,20S TAS TAS0

C26,20R+C27,20S TAS TAS1

C28,20S TAS TAS2

C27,20R TAS TAS3

C28,20R TAS TAS4

C29,20S TAS TAS5

C29,20R TAS TAS6

Mono-aromatic Steroids 5b(H)-C27 (20S) MAS+ MAS1

5b(H)-C27 (20R) MAS+ MAS2

4-12 Attachment A

2020 LTEMP Report

54


5a(H)-C27 (20S) MAS MAS3

5b(H)-C28 (20S) MAS+ MAS4

5a(H)-C27 (20R) MAS MAS5


5b(H)-C28 (20R) MAS+ MAS7


5b(H)-C29 (20S) MAS+ MAS8



5b(H)-C29 (20R) MAS+ MAS11


Spill oil from 2017

0

50

100

150

200

250

300

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9T1

0 TsTR

30

aTR

30

bTm 14

a1

4b

T15

T16

T17

T18

OL

T19

T20

T21

T22

Gam

mT2

6T2

7T3

0T3

1T3

2T3

3T3

4T3

5 S4 S5 S8D

IA2

9S

DIA

29

RS1

8S1

9S2

0S2

4S2

5S2

8S1

4S1

5S2

2S2

3S2

6S2

7P

reg

MP

reg

EPre

gAEP

regB

TAS0

TAS1

TAS2

TAS3

TAS4

TAS5

TAS6

MA

S1M

AS2

MA

S3M

AS4

MA

S5M

AS6

MA

S7M

AS8

MA

S9M

AS1

0M

AS1

1M

AS1

2

ng

/g

BAR-1709-1

4-12 Attachment A

2020 LTEMP Report

55

APPENDIX 2. Analytic Results for 2020 Field Samples and Blanks

2020 AMT Sediments (PAH, SHC, and Biomarkers)

The dotted red line represents the sample-specific method detection limits (MDL) for PAH, SHC, and biomarkers. The biomarker, hopane (T19; highlighted in

gold), is used for scaling reference overlay patterns.

0

2

4

6

8

10

12

14

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-20-2-1

TPAH182

Hopane15

11-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

AMT-S-20-2-1

TSHC3

0

2

4

6

8

10

12

14

16

18

20

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-S-20-2-1

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-20-2-2

TPAH122

Hopane15

11-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

AMT-S-20-2-2

TSHC2

0

5

10

15

20

25

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-S-20-2-2

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-S-20-2-3

TPAH88

Hopane19

11-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

AMT-S-20-2-3

TSHC2

0

5

10

15

20

25

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-S-20-2-3

4-12 Attachment A

2020 LTEMP Report

56

2020 GOC Sediments (PAH, SHC, and Biomarkers)

The dotted red line represents the sample-specific method detection limits (MDL) for PAH, SHC, and biomarkers.

0

0.5

1

1.5

2

2.5

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-20-2-1

TPAH24

Hopane2

12-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GOC-S-20-2-1

TSHC2

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-S-20-2-1

0

0.5

1

1.5

2

2.5

3

3.5

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-20-2-2

TPAH34

Hopane3

12-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GOC-S-20-2-2

TSHC2

0

1

2

3

4

5

6

7

8

9

10

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-S-20-2-2

0

0.5

1

1.5

2

2.5

3

3.5

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-S-20-2-3

TPAH34

Hopane3

12-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GOC-S-20-2-3

TSHC1

0

1

2

3

4

5

6

7

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR30

bTm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-S-20-2-3

4-12 Attachment A

2020 LTEMP Report

57

2020 AMT Tissues (PAH, SHC, and Biomarkers)


0

2

4

6

8

10

12

14

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-B-20-2-1

TPAH66Hopane0

11-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

1.4

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

AMT-B-20-2-1

TSHC5

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-B-20-2-1

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-B-20-2-2

TPAH66Hopane0

11-Jun-20

0

0.5

1

1.5

2

2.5

3

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

AMT-B-20-2-2

TSHC6

0

0.5

1

1.5

2

2.5

3

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-B-20-2-2

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

AMT-B-20-2-3

TPAH55Hopane0

11-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

AMT-B-20-2-3

TSHC5

0

0.5

1

1.5

2

2.5

3

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

AMT-B-20-2-3

4-12 Attachment A

2020 LTEMP Report

58

2020 JAP Tissues (PAH, SHC, and Biomarkers)


0

2

4

6

8

10

12

14

16

18

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAP-B-20-2-1

TPAH98Hopane7

11-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

JAP-B-20-2-1

TSHC5

0

1

2

3

4

5

6

7

8

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

JAP-B-20-2-1

0

10

20

30

40

50

60

70

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAP-B-20-2-2

TPAH158Hopane42

11-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

JAP-B-20-2-2

TSHC4

0

5

10

15

20

25

30

35

40

45

50

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

JAP-B-20-2-2

0

2

4

6

8

10

12

14

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAP-B-20-2-3

TPAH73Hopane5

11-Jun-20

0

0.5

1

1.5

2

2.5

3

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

JAP-B-20-2-3

TSHC6

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

JAP-B-20-2-3

4-12 Attachment A

2020 LTEMP Report

59

2020 GOC Tissues (PAH, SHC, and Biomarkers)


0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-B-20-2-1

TPAH59Hopane0

12-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GOC-B-20-2-1

TSHC4

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-B-20-2-1

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-B-20-2-2

TPAH49Hopane4

12-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GOC-B-20-2-2

TSHC4

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-B-20-2-2

0

2

4

6

8

10

12

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GOC-B-20-2-3

TPAH49Hopane0

12-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GOC-B-20-2-3

TSHC4

0

0.5

1

1.5

2

2.5

3

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GOC-B-20-2-3

4-12 Attachment A

2020 LTEMP Report

60

2020 Tissue Lab Method Blanks


0

0.1

0.2

0.3

0.4

0.5

0.6

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

WG1389790-1-BLANK

TPAH2

Hopane#N/A

0-Jan-00

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

#N/A

WG1389790-1-BLANK

TSHC-

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

#N/A

WG1389790-1-BLANK

0

0.1

0.2

0.3

0.4

0.5

0.6

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

WG1390678-1-BLANK

TPAH2

Hopane#N/A

0-Jan-00

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

#N/A

WG1390678-1-BLANK

TSHC-

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

#N/A

WG1390678-1-BLANK

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

#N/A

WG1392670-1-BLANK

TPAH#N/A

Hopane0

0-Jan-00

0

0.01

0.02

0.03

0.04

0.05

0.06

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

WG1392670-1-BLANK

TSHC0

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

WG1392670-1-BLANK

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

#N/A

WG1393174-1-BLANK

TPAH#N/A

Hopane0

0-Jan-00

0

0.01

0.02

0.03

0.04

0.05

0.06

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

WG1393174-1-BLANK

TSHC0

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

WG1393174-1-BLANK

4-12 Attachment A

2020 LTEMP Report

61

2020 Sediment Lab Method Blanks


0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

WG1386048-1-BLANK

TPAH0Hopane#N/A

0-Jan-00

0

0.01

0.02

0.03

0.04

0.05

0.06

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

WG1386048-1-BLANK

TSHC0

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

#N/A

WG1386048-1-BLANK

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

WG1389307-1-BLANK

TPAH1Hopane#N/A

0-Jan-00

0

0.005

0.01

0.015

0.02

0.025

0.03

0.035

0.04

0.045

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

WG1389307-1-BLANK

TSHC0

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

#N/A

WG1389307-1-BLANK

4-12 Attachment A

2020 LTEMP Report

62

2020 Red Tissues (Valdez Small Boat Harbor Entrance)


0

50

100

150

200

250

300

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

RED-B-20-2-1

TPAH1139Hopane15

8-Jun-20

0

0.2

0.4

0.6

0.8

1

1.2

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

RED-B-20-2-1

TSHC7

0

5

10

15

20

25

30

35

40

45

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

RED-B-20-2-1

0

50

100

150

200

250

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

RED-B-20-2-2

TPAH977Hopane14

8-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

RED-B-20-2-2

TSHC5

0

2

4

6

8

10

12

14

16

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

RED-B-20-2-2

0

50

100

150

200

250

300

350

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

RED-B-20-2-3

TPAH1273Hopane13

8-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

RED-B-20-2-3

TSHC5

0

2

4

6

8

10

12

14

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

RED-B-20-2-3

4-12 Attachment A

2020 LTEMP Report

63

2020 Green Tissues (Valdez Small Boat Harbor Entrance)


0

20

40

60

80

100

120

140

160

180

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GRN-B-20-2-1

TPAH763Hopane15

8-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GRN-B-20-2-1

TSHC5

0

2

4

6

8

10

12

14

16

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GRN-B-20-2-1

0

50

100

150

200

250

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GRN-B-20-2-2

TPAH916Hopane25

8-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GRN-B-20-2-2

TSHC5

0

5

10

15

20

25

30

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GRN-B-20-2-2

0

20

40

60

80

100

120

140

160

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GRN-B-20-2-3

TPAH712Hopane18

8-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GRN-B-20-2-3

TSHC5

0

2

4

6

8

10

12

14

16

18

20

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GRN-B-20-2-3

4-12 Attachment A

2020 LTEMP Report

64

2020 Jack Bay Tissues (PAH, SHC and Biomarkers)


0

1

2

3

4

5

6

7

8

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAK-B-20-2-1

TPAH36Hopane0

9-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

JAK-B-20-2-1

TSHC3

0

0.5

1

1.5

2

2.5

3

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

JAK-B-20-2-1

0

1

2

3

4

5

6

7

8

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAK-B-20-2-2

TPAH34Hopane0

9-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

JAK-B-20-2-2

TSHC3

0

1

2

3

4

5

6

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

JAK-B-20-2-2

0

1

2

3

4

5

6

7

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

JAK-B-20-2-3

TPAH35Hopane5

9-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

JAK-B-20-2-3

TSHC3

0

1

2

3

4

5

6

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

JAK-B-20-2-3

4-12 Attachment A

2020 LTEMP Report

65

2020 Galena Bay Tissues (PAH, SHC and Biomarkers)


0

1

2

3

4

5

6

7

8

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GAL-B-20-2-1

TPAH27Hopane0

9-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GAL-B-20-2-1

TSHC3

0

0.5

1

1.5

2

2.5

3

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GAL-B-20-2-1

0

1

2

3

4

5

6

7

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GAL-B-20-2-2

TPAH27Hopane0

9-Jun-20

0

0.05

0.1

0.15

0.2

0.25

0.3

0.35

0.4

0.45

0.5

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GAL-B-20-2-2

TSHC2

0

0.5

1

1.5

2

2.5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GAL-B-20-2-2

0

1

2

3

4

5

6

7

DEC

DEC

1D

EC2

DEC

3D

EC4

BT0

BT1

BT2

BT3

BT4 N

0N

1N

2N

3N

4B

PD

BF

AC

YA

CN F F1 F2 F3

AN

TP

HN

PA

1P

A2

PA

3P

A4

RET

DB

T0D

BT1

DB

T2D

BT3

DB

T4C

AR

BB

BF

FLA

PYR FP

1FP

2FP

3FP

4N

BT

NB

T1N

BT2

NB

T3N

BT4

BA

A C0

C1

C2

C3

C4

BB

FB

JKF

BA

FB

EPB

AP

PER IP

DA

HA

BG

HIP

ng

/g

GAL-B-20-2-3

TPAH59Hopane0

9-Jun-20

0

0.1

0.2

0.3

0.4

0.5

0.6

C9

C1

0C

11

C1

2C

13

138

0C

14

147

0C

15

C1

6N

pri

sC

17

Pri

sC

18

Ph

yC

19

C2

0C

21

C2

2C

23

C2

4C

25

C2

6C

27

C2

8C

29

C3

0C

31

C3

2C

33

C3

4C

35

C3

6C

37

C3

8C

39

C4

0

ug

/g

GAL-B-20-2-3

TSHC2

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

T4 T5 T6 T6a

T6b

T6c

T7 T8 T9 T10 Ts

TR3

0a

TR3

0b

Tm 14a

14b T1

5T1

6T1

7T1

8O

LT1

9T2

0T2

1T2

2G

amm

T26

T27

T30

T31

T32

T33

T34

T35 S4 S5 S8

DIA

29S

DIA

29R

S18

S19

S20

S24

S25

S28

S14

S15

S22

S23

S26

S27

Pre

gM

Pre

gEP

regA

EPre

gBTA

S0TA

S1TA

S2TA

S3TA

S4TA

S5TA

S6M

AS1

MA

S2M

AS3

MA

S4M

AS5

MA

S6M

AS7

MA

S8M

AS9

MA

S10

MA

S11

MA

S12

ng

/g

GAL-B-20-2-3

4-12 Attachment A

2020 LTEMP Report

66

APPENDIX 3. PROJECT HISTORY The Long-Term Environmental Monitoring Program (LTEMP) data serve to monitor and provide independent quality-

control for Alyeska Pipeline Service Company’s Valdez Marine Terminal (VMT) and tanker operations throughout the

Prince William Sound (PWS) and Gulf of Alaska (GOA) region. The primary goal of this on-going Prince William Sound

Regional Citizens’ Advisory Council (PWSRCAC) program is to monitor impacts from oil transportation activities on

the environment at selected sites from PWS and GOA for “as long as the oil flows through the pipeline.”

At the VMT, the Ballast Water Treatment Facility (BWTF) treats and discharges oil-contaminated ballast water

offloaded from tankers utilizing the terminal. Since the Program’s inception in 1993, two stations have been

traditionally sampled to assess impacts from the effluent: at Alyeska’s terminal adjacent to the offshore BWTF

discharge diffusers near Berth 4 for sediments (AMT-S) and at Saw Island near Berth 5 for mussels (AMT-B); and at

Gold Creek (GOC), a reference station 6 km across the Port (Figure 28) for both sediments and mussels. Another

station, Jackson Point (JAP), was added in 2016 near Berth 3, towards the opposite (eastern) end of the terminal.

Currently measured variables include levels of polycyclic aromatic hydrocarbons (PAH) and saturated hydrocarbons

(SHC), as well as oil biomarkers in mussel (Mytilus trossulus) tissues from the three stations within the Port. Eight

additional stations, comprising the geographic reach of the Exxon Valdez oil spill (EVOS), are now sampled every five

years (last sampled in 2018) between Valdez and Kodiak (Figure 29, Table 6).

Figure 28. LTEMP sampling stations in Port Valdez adjacent to (AMT-B, AMT-S, and JAP) and 6 km northwest (GOC) of the VMT.

This Google Earth image shows a tanker docked at Berth 5.

D

B – Tissue



S – Sediment grab

S

Valdez

Gold Creek (GOC)

Saw Island

Jackson Point (JAP)


S

AMT

B/P

B/P B/P

(Jack and Galena Bay off map)

Red & Green

2020 Spill

B B

4-12 Attachment A

2020 LTEMP Report

67

Figure 29. Map of the LTEMP sites with station abbreviations.

Sediment samples collected from the two Port stations are analyzed for PAH, SHC, particle grain size, and total

organic carbon content, with oil biomarkers added in recent years to confirm petrogenic sources. Sampling and

analytical methods are modelled after the protocols developed by the National Oceanic and Atmospheric

Administration (NOAA) Status and Trends Mussel Watch Program as fully detailed in previous annual monitoring

reports prepared by Kinnetic Laboratories, Inc. (KLI), the Geochemical and Environmental Research Group (GERG),

and Payne Environmental Consultants, Inc. (PECI).

Following the first five years of the program, the collective results from the KLI/GERG team were reviewed in a

synthesis paper (Payne et al., 1998). At that time, sampling was more extensive, and identification of weathered

sources was important (Table 5-1 in Payne et al., 1998). The results effectively documented higher background oil

levels while identifying hot spots and both large and small spill events. Subsequently, the PWSRCAC reduced the

scope of the program from triannual to just spring and summer sampling of regional mussel tissues and Port Valdez

sediments. Fall mussel sampling (without sediments) was then re-added just in Port Valdez (AMT-B and GOC-B) to

better track the terminal’s discharge. Mussel-tissue SHC analyses that were dropped from the original program in

1995 (due to results being confounded by lipid interference) were reinstated in 1998 using improved laboratory

methods.

4-12 Attachment A

2020 LTEMP Report

68


spring, summer, autumn (SSA); or summer only (S). Sediments (not shown) were sampled in spring and summer at

AMT-S and GOC-S from 1993-2008, and afterwards only in summer.

LTEMP Station Mussel Samplings

Port Valdez Prince William Sound Gulf of Alaska

AMT-B JAP GOC-B KNH DII SLB ZAI SHB COH AIB WIB SHH


1994 SS SS SA SA SA SA SA SA SA SA












2006 SSA SSA SS SS SS SS SS SS SS SS SS


2008 SA SSA S S S S S S S S

2009 SS SS S S S S S S S S

2010 S S S

2011 S S S

2012 S S S

2013 S S S S S S S S S S

2014

2015 S S S S

2016 S S S

2017 S S S

2018 S S S S S S S S S S S

2019 S S S

2020 S S S

In 2001, another comprehensive data evaluation and synthesis was completed on just the Port Valdez sites’ LTEMP

results (Payne et al., 2001). From AMT-B and the GOC-B reference site, Alaska North Slope (ANS) crude oil residues

from the terminal’s BWTF were shown to accumulate in the intertidal mussels. However, the sediment and tissue

(and the estimated water-column) PAH and SHC levels were very low.

More importantly, the pattern and trend of signatures suggested a novel transport/exposure mechanism;

discrimination of particulate (oil droplet) and dissolved-phase signals in the water column correlated with seasonal

4-12 Attachment A

2020 LTEMP Report

69

uptake in Port Valdez mussels (Payne et al., 2001). Stratified waters formed during the milder seasonal winds of late-

spring/summer kept the discharge plume’s particulate oil-phase droplets from the surface but dissolved-phase

components could be observed in the intertidal mussels. The wind-mixed, non-stratified waters of fall/winter

allowed some portion of the plume’s particulate/oil phase to surface, which was then visible in the mussel tissues.

The results suggested a surface microlayer mechanism may be responsible for seasonal transport of ANS weathered

oil residues from the BWTF diffuser to intertidal zones across the fjord. Combined with other study results showing

toxic absorption in herring eggs at trace levels, the authors warned that the potential for photo-enhanced toxicity

of concentrated contaminants in a surface microlayer should be considered in future impact investigations (Payne

et al., 2001, 2003a, 2003b, 2005c). This effect has likely disappeared with the current diminished discharge flows.

In July 2002, PECI and the NOAA/National Marine Fisheries Service (NMFS) Auke Bay Laboratory (ABL) began

collecting and analyzing the LTEMP samples. Changing laboratories can be problematic; detailed discussions of the

transitional 2002/2003 LTEMP samples and inter-laboratory comparisons of split samples and Standard Reference

Materials (SRMs) analyzed by both GERG and ABL are presented in Payne et al., (2003b). The results from the

2003/2004 LTEMP and a comprehensive review and synthesis of all analyses completed since the beginning of the

program are available in Payne et al., (2005a, 2006, 2008a). Results and discussion of the program through 2006

were also published in the journal, “Marine Pollution Bulletin” (Payne et al., 2008b).

The program again changed analytical services in 2016 when ABL closed its hydrocarbon facilities in Juneau, Alaska.

This decision necessitated a switch to Alpha/NewFields in Mansfield, Massachusetts as the PWSRCAC contract

laboratory for LTEMP. Alpha/NewFields was the primary laboratory used by NOAA and other State and Federal

Trustees for the 2010-2016 Natural Resource Damage Assessment (NRDA) effort following the BP Deepwater Horizon

oil spill (Driskell and Payne, 2018a, 2018b; Payne and Driskell, 2017a, 2018a; Stout and Payne, 2016a, 2016b, 2017;

Stout et al., 2016a, 2016b). For that event, Alpha/NewFields analyzed ~30 thousand sediment, water, and tissue

samples, all with independent, third-party quality control (QC) validation as part of that litigation-sensitive effort.

The LTEMP lab transition also involved performance-based round-robin intercalibration programs in which both ABL

and Alpha/NewFields participated to demonstrate they were generating comparable data with known precision,

accuracy, method detection limits, and representativeness.

Prior to this report, all 10 LTEMP sites were most recently visited in July 2008, April 2009, and then, beginning the

reduced effort, 5-year interval scheme, in July 2013 and 2018 (Table 6). Visits included three sites in or near the Port

to monitor terminal and tanker operations, six others to monitor the more remote sites for lingering EVOS impacts,

plus Sheep Bay (SHB) which serves as a non-EVOS-impacted control. Initially, to more thoroughly monitor Port

operations, LTEMP collections had been taken tri-annually at the two Port sites and nearby Knowles Head (KNH) but

efforts were later reduced to annual sampling. Under this modified plan, in 2015, sampling occurred at four of the

10 LTEMP stations: AMT, GOC, KNH, and SHB (Payne et al., 2016). In 2016, another mussel sampling site was added

at JAP at the terminal but on the opposite side (east) of the active berths and the traditional station at Saw Island

(AMT-B). This site was meant to evaluate a potential PAH gradient to either side of the BWTF outfall and to correlate

tissue data with passive-sampling devices (PSDs) that were concurrently deployed subsurface at the same terminal

locations (Minick and Allan, 2016; Allan, 2018).

Sampling in 2018 fell on the five-year cycle and covered all 11 sites (Payne and Driskell, 2019). In 2019-20, just the 3

Port Valdez sites were visited (Payne and Driskell, 2020).

Recent years have brought change to both the system and the environment as pipeline throughput has dropped

from 85.3 million gallons per day (MGD) at its peak in 1988 to current levels of 20.2 MGD (Figure 30). Likewise, tanker

regulations have instituted double-hulled tankers with segregated ballast. Aboard segregated-ballast vessels, empty

4-12 Attachment A

2020 LTEMP Report

70

cargo tanks are typically used for supplemental ballast only when operationally necessary (e.g., during winter

storms), (i.e., the normal segregated ballast waters are uncontaminated seawater that do not require treatment for

hydrocarbons). Treated-ballast water discharges to the Port have also swung from a maximum of around 15 MGD in

1990 to currently only 0.96 MGD (Table 7). Facility operators estimate, and Alyeska’s Discharge Monitoring Report

(DMR) data confirm, that more than half of the current BWTF effluent discharge in summer is from the terminal’s

stormwater runoff (Rich Loftin, personal communication, 2016). In summary, less tanker traffic, cleaner ballast, and

an improved ballast-water-treatment configuration at the VMT have resulted in substantial reductions in detected

hydrocarbon concentrations and composition in the field samples. All discharges are made under an Alaska Pollutant

Discharge Elimination System (APDES) Permit for which the PWSRCAC submitted detailed reviews during the last

three permit renewal cycles (Payne et al., 2003c and 2012, PWSRCAC, 2019).

Figure 30. Annual pipeline oil throughput (M barrels) from Alyeska statistics.

Table 7. Recent average daily throughput of Alyeska Pipeline and ballast water treatment (MGD).

2015 2016 2017 2018 2019 2020

Pipeline throughput 20.6 21.8 22.3 21.4 20.6 20.2

BWTF throughput 1.10 1.00 0.88 1.11 0.81 0.96

APPENDIX 4. METHODS Collection and analytical methods have been described in previous LTEMP reports (Payne et al., 2003b, 2005a, 2006,

2008a, 2010a, 2013, 2015, 2016; Payne and Driskell 2017b, 2018c, 2019). Briefly, three replicates of 30 mussels are

collected by hand at each site while triplicate sediment samples are collected from the two Port locations (AMT-S

and GOC-S) using a modified Van Veen grab (Figure 31). Sampling protocols have remained the same but as noted

in Appendix 3, Alpha Analytical Laboratory (Mansfield, MA) under the guidance of NewFields Environmental

Forensics Practice (Rockland, MA) now provides the analytical services.

0

100

200

300

400

500

600

700

800

1977

1979

1981

1983

1985

1987

1989

1991

1993

1995

1997

1999

2001

2003

2005

2007

2009

2011

2013

2015

2017

2019

2021

Mill

ion

bar

rels

Total Pipeline Oil Throughput

4-12 Attachment A

2020 LTEMP Report

71

Figure 31. Deploying the Van Veen grab (upper left). View of benthic sediments collected with intact surface layer

(and residual water) in jaws (upper right) before sampling (bottom). Work photos courtesy of David Janka.

The usual hydrocarbon data are reported: polycyclic aromatic hydrocarbons (PAH), sterane/triterpane biomarkers

(S/T), and saturated hydrocarbons (SHC). Semi-volatile compounds, the PAH, alkylated PAH, and petroleum

biomarkers, are analyzed using selected ion monitoring gas chromatography/mass spectrometry (SIM GC/MS) via a

modified Environmental Protection Agency (EPA) Method 8270 (aka 8270M). This analysis provides the

concentration of 1) approximately 80 PAH, alkylated PAH homologues, individual PAH isomers, and sulfur-containing

aromatics and 2) approximately 50 tricyclic and pentacyclic triterpanes, regular and rearranged steranes, and

triaromatic and monoaromatic steroids. Complete lists of PAH, SHC, and biomarker (S/T) analytes are presented in

Appendix 1 along with the analyte abbreviations used in figures throughout this report.

Using a modified EPA Method 8015B, SHC in sediments and tissues are quantified as total extractable materials

(TEM; C9-C44) and as concentrations of n-alkanes (C9-C40) and selected (C15-C20) acyclic isoprenoids (e.g., pristane and

phytane). A high-resolution gas chromatography-flame ionization detector (GC/FID) fingerprint of the sediment and

tissue samples is also provided.

Added to the project in 2017, low-density polyethylene, PSD were deployed for ~30 days following Oregon State

University (OSU) protocols (Figure 32) (Sowers et al., 2008, Huckins et al., 2006, O’Connell et al., 2013). Laboratory

handling, sample extraction and analyses of the PSDs followed respective OSU, Food Safety and Environmental

Stewardship (FSES) Program Standard Operating Procedures (SOPs).

4-12 Attachment A

2020 LTEMP Report

72

Figure 32. Example of passive sampling device (PSD) consisting of a low-density polyethylene membrane strip

enclosed in a stainless-steel container and deployed subsurface in near-shore subtidal waters adjacent to LTEMP

mussel collection sites. Work photos courtesy of David Janka.

DATA ANALYSIS

Port Valdez LTEMP data are interesting in a couple of respects; the data are quite rich in number of analytes and

long in time span but only represent three locations in the Port. From these data, there are two main questions: 1)

do the detected hydrocarbons come from the ballast-water operations and 2) do the hydrocarbons reach a level for

concern?

There have been three approaches to addressing these questions. Initially, the PWSRCAC contractors, Kinnetic

Laboratories, Inc. (KLI), used hypothesis-testing statistical methods (e.g., ANOVAs and t-tests) to see whether various

values, ratios, and indices were significantly different in order to highlight trends. University of Alaska Fairbanks

(UAF) contractors for Alyeska tend towards exploratory multivariate approaches (e.g., PCA, MDS, kriging) on various

values to understand sources, correlations, and trends. The results are impressive with a much larger set of stations

sampled but considering the subset of PAH analytes they report, which constrains any fuller understanding, the

project leaves many questions unanswered. In contrast, since the LTEMP data set represents only three Port stations,

we emphasize expert pattern recognition to tease out/confirm the source and phase state of the sample and then

apply those results for the Port’s environmental trends scenario.

Per common practice, analytical chemistry results are presented in this report as bar plot profiles for each analyte.

Note that for the alkylated PAH (whereby C1-, C2-, C3- and C4- are the alkyl members, meaning carbons attached to

the parent molecule), the plotted bars are actually histograms representing the sum of alkylated homologue

components. For example, C1-naphthalene, a two-ringed compound with an attached methyl group (one carbon)

has two isomer forms with each appearing as a unique peak on the instrument. They are individually quantified but

4-12 Attachment A

2020 LTEMP Report

73

reported and plotted as a combined sum (abbreviated as “N1”); C2-naphthalenes (with two attached methyl groups

or one ethyl group) have 12 isomers (abbreviated as “N2”), and C4-chrysenes have 1,016 possible isomers

(abbreviated as “C4” when plotted). Again, these alkylated isomer homologues are summed into one value for the

bar plots. Also, as described in each sample’s figure caption in this report, some appropriate reference is depicted

as a red-colored reference line scaled and overlaid on the selected individual tissue or sediment’s profile (e.g., the

summer BWTF-effluent profile or ANS source oil). Method detection limit (MDL) overlays are commonly shown for

selected time-series data profiles and are shown for all 2020 samples in Appendix 2. Details of our BWTF-effluent

sampling and analytical procedures and the importance of differentiating between dissolved and particulate/oil

phases are published in Appendix 2 of Payne and Driskell (2017b).

BIOMARKERS

Petroleum biomarkers are conservative, weathering-resistant, hydrocarbon compounds, unique to each oil

formation, which present a less degraded hydrocarbon signature than those of the SHC and PAH. In the environment,

the SHC are quickly consumed by microbes, which gives forensic reviewers a perspective of “freshness” of crude oil

patterns while also distinguishing petroleum distillates (diesel vs. fuel oil vs. crude oil, etc.), and tagging contributions

from other (primarily biogenic) sources. PAH compounds are more persistent, weathering slower in predictable

patterns and rates, which serve to track a longer-term fate, behavior, and mixing with other sources. In contrast, the

biomarkers are the hydrocarbon “tattoos,” enduring telltales of oil’s presence even as the PAH and SHC patterns are

weathering and disappearing. For LTEMP, biomarkers can facilitate and confirm detection of VMT-derived, ANS

crude-oil constituents in Port sediments – even when the PAH are heavily degraded. 2

Beginning in 2011, biomarkers were analyzed for sediments and in 2016, also for tissues. For routine monitoring,

however, mussel tissue biomarkers are generally less effective than in sediments as the mussels regularly purge and,

currently at most LTEMP stations, only carry trace-level, dissolved-phase, and combustion-derived PAH components

(i.e., no biomarkers). Note that the biomarkers are water-insoluble and thus, would only be detected in tissues when

particulate oil was present, for example, free oil droplets from a chronic or recent release such as the 2020 VMT

sump spill (report in prep).

Multiple approaches have been suggested for interpreting biomarker data, but some degree of expert-guided

pattern matching must be employed. Most schemes involve various diagnostic ratios (Stout and Wang, 2016) with

several ratios normalized to the highly conservative 17α(H),21β(H)-hopane (also labeled T19 or C30 hopane and

herein tagged in the bar plots with a golden fill color for visual reference). But despite the purported persistence of

biomarkers, depending on the local environs and microbial adeptness, all ratios are not equally effective and must

be individually evaluated for a given spill/habitat. For interpreting LTEMP data, we initially screen PAH and biomarker

results graphically with an ANS-oil-reference overlay normalized to the sample’s hopane. For biomarkers, the

frequently reliable, Ts/Tm and norhopane (T15)/hopane ratios were used to confirm the visual similarities. For this

report, we utilize all appropriate and available ratio data and present the overall patterns to facilitate their

interpretations.

2 For readers who are not familiar with oil spill fingerprinting or forensics, see Appendix 6 in our 2015 LTEMP Report (Payne et al., 2015) for a background primer specific to ANS crude oil, combustion products, and other potential oil sources in Port Valdez and the PWS/GOA region. Additional details are available in Stout and Wang (2007 and 2016).

4-12 Attachment A

2020 LTEMP Report

74

APPENDIX 5. LABORATORY PERFORMANCE

LABORATORY QUALITY CONTROL All Alpha/NewFields-analyzed constituents (Appendix 1) are reported on a ng/g dry weight (DW) basis uncorrected

for blanks or surrogate recoveries. Surrogates are novel or deuterated compounds added in known amounts to each

raw sample to assess, by their final percent recovery, the efficiency of extraction and analysis. Surrogate recoveries

are considered acceptable if they are between 40% and 120%. A single recovery deviance flags the sample with

cautionary remarks; multiple recovery deviations would require batch reanalysis. Surrogate recovery standards were

met for all PAH, biomarker and alkane surrogate hydrocarbons analyzed during the 2020 reporting period (Table 8).

Laboratory method blanks for each analytic sample batch demonstrated (sub-ng/g) laboratory or analytic

background PAH interference.

Table 8. Surrogate recovery statistics by matrix from 2020 Alpha Laboratory analyses.

Tissue Sediment

Surrogate Average (%) Max Min Count StdDev Average (%) Max Min Count StdDev

5B(H)Cholane 97 109 87 25 6 94 100 89 15 3

Benzo[a]pyrene-d12 83 94 73 29 5 95 102 87 15 5

d50-Tetracosane 91 101 81 24 5 93 102 85 15 4

Naphthalene-d8 64 76 55 29 6 82 95 68 15 7

o-terphenyl 93 96 85 15 3

Phenanthrene-d10 80 91 74 29 3 99 108 90 15 4

Mussel-tissue hydrocarbon levels are often so low throughout the study region that individual PAH were reported

at below-MDL concentrations and patterns in both tissues and their associated laboratory/method blanks. At these

exceptionally low PAH levels, it is not possible to assure that the measured analytes in the field samples accurately

quantify the analytes (discussed in next section).

In addition, some matrix interferents were flagged for two sediment biomarkers, T26 and T32, and one SHC, C32, in

their lab qualifiers. As obvious anomalies, they were ignored for sediment data-pattern interpretations. Per our

standard forensic reporting practices, the data discussed herein are neither blank-corrected nor surrogate-recovery-

corrected but are simply reported as raw data (with below-MDL values flagged as estimated). All PAH, biomarker,

and SHC profiles presented in Appendix 2 are shown with their analytical-batch-associated method blanks along with

sample-specific MDLs overlaid on the histogram profiles.

METHOD DETECTION LIMITS

One lab-performance quality control (QC) measure is the EPA-formulated, statistically-derived, analyte-specific,

Method Detection Limit (MDL) that EPA defines as “the minimum measured concentration of a substance that can

be reported with 99% confidence that the measured concentration is distinguishable from method blank results.”

Alpha Analytical Laboratory’s MDLs for hydrocarbons exceed the performance of most commercial labs, falling

within the accepted stricter levels for forensic purposes (Table 9).

4-12 Attachment A

2020 LTEMP Report

75

Table 9. Alpha Analytical MDL target ranges.

Analytes

Sediment

(30 g, sample size)

Tissue

(15 g sample size)

Water

(1 L sample size)

Oil Reporting

Level (RL)

PAH and biomarkers 0.1-0.5 ng/g DW 0.2-1.0 ng/g DW 1-5 ng/L 2.0 µg/g

SHC 0.05 µg/g DW 0.01-0.08 µg/g DW 0.8 µg/L 200 µg/g

For data interpretation, there are generally two approaches on the use of MDLs: 1) censor all below-MDL data to

some pre-decided level (which leads to further issues on how to interpret partially censored, multi-analyte data sets

such as LTEMP) or 2) treat below-MDL data as estimated real values. For reasons described below, it is felt that the

second option best serves the purpose of the LTEMP program. For both the readers and our benefit in reviewing

data, individual analyte MDLs (adjusted to sample weight) appear as red dotted lines on PAH and SHC plots where

appropriate in report figures and in all sample plots presented in Appendix 2.3

By definition, EPA’s MDL protocol is designed to control against false positives at the 99% confidence level in an ideal

matrix. In other words, MDLs are meant to represent a trustworthy value of low detection, below which, due to

expected uncontrolled factors, lower results are not as reliable—the values are estimates of lesser confidence. This

reporting bulwark is certainly required when reviewing a crucial single-analyte result (e.g., water arsenic

concentrations, where the statistically determined MDL value ensures against toxic consequences). But there are

two differences between this example and the LTEMP dataset.

First, there are no “critical values” involved in the current LTEMP data review; false positives will not affect the

overall findings of “PAH are dropping to lower historic lows.” While the MDL procedure is designed to avoid false

positives at the 99% confidence level; if a lower confidence level is acceptable, then EPA-defined MDL levels are

unnecessarily stringent for the application.

Secondly, because LTEMP data interpretations are based on multi-analyte patterns rather than single values,

additional confidence accrues from “pattern expectations.” Conceptually, the more information known about a

system or data set, the higher the confidence when seeing recognizable patterns. Such is the case with LTEMP data.

Oil weathers predictably (see Appendix 6 in Payne et al., 2015) and if a sample’s PAH profile appears to represent a

recognizable pattern, then applying the statistically established, single-analyte MDLs to censure the data would be

unnecessarily conservative. For example, if a sample’s phenanthrenes/anthracenes (P/As) were reported above MDL

levels while dibenzothiophenes and chrysenes (DBTs and Cs) were reported below MDLs but in the same pattern

and ratio as the source oil, there would be sufficient confidence in the expected patterns that those detected

analytes, albeit below-MDL, were not false positives and that the values had been reasonably estimated. In past

LTEMP efforts where near-trace-level tissue data were reported, this added-confidence was further bolstered by

seeing higher-level coincidence patterns of within-site fidelity and regional-wide commonalities that collectively

changed between years—which could only occur if the patterns were real, reflecting conditions in the field, and not

false positives from lab or procedural artifacts (e.g., see Appendix 3 in Payne et al., 2015). In LTEMP data, MDLs

mainly serve to tag when reported values have become, to some degree, estimated. Conversely, when an

unrecognizable pattern or anomalous spike appears, it is easily spotted, flagged as an outlier, and closely examined

along with any corroborating evidence (e.g., lab QC and field notes) to see if it makes any sense or is indeed a lab,

sampling, or field anomaly.

3 For forensics evaluations, PAH, SHC, and S/T plots in the main body of this report are typically shown with dotted-red-line overlays corresponding to a relevant reference source oil or BWTF effluent.

4-12 Attachment A

2020 LTEMP Report

76

In the 2018 and 2019 mussel-tissue samples, however, similar trace-level, below-MDL PAH patterns were observed

in all the field samples and their batch-associated laboratory method blanks. With these PAH data, source-

identifications based on pattern recognition are not possible; it can only be reported that the individual and total

PAH (TPAH) concentrations are below a background level. In 2020, concentrations rose above MDLs.

APPENDIX 6. SEDIMENT GRAIN SIZE The LTEMP sediment-sampling locations comprise a heterogeneous slope at AMT-S and sediment shelf at GOC-S

inside a fjord dynamically swept by tidal currents (and prop wash at AMT-S). Plus, with LTEMP sampling guided by

GPS, the sites have been accruing grab-sampler pock marks and drag scars at the same locations for 20 years.

Reassuringly, when the sampling vessel gets off-site at GOC-S, we begin to see gravel tell-tales in the grab. The grain-

size component trends are presented here with only modest confidence considering the non-rigorous collection

methods (i.e., spooning up 250 mL of sediment remnants after collecting the less consolidated, top cm of surface

floc for hydrocarbon analyses). There was also a change in analytical labs in 2016.

Sediment particle-grain-size (PGS) samples are presented for all 2006-2020 collections in two formats: the standard

cumulative (%) grain-size curves and a 3D trend plot. For this project, the grain-size data only serve to demonstrate

the long-term constancy and comparability of the sampling site environs.

Both sites are dominated by glacial flour inputs, showing approximately equal portions of clay and silt with minor

sand components (Figure 33 and Table 10) and with both sites showing minor trends and outliers. In the 3D plots

(Figure 34), note there are annual shifts at GOC-S (~30m depth) to higher sand content and back (albeit still a minor

component, mostly ~10%) and with a return to earlier conditions in 2013. But in 2015, a shift to coarser particles

occurred when silt increased and clay decreased dramatically. In 2016, 2017, and 2018, there was further return

towards a clay- and slit-dominated substrate. The 2019 GOC-S samples have remained close to the average portions.

At AMT-S (~70 m depth), there has been a cycle of increasing clay content through 2009 and then a decrease,

returning to 2006 levels by 2015-16. In 2016, there was also a halving of sand, albeit still a minor component (~3%).

In 2017, the sand portion decreased further as clay became dominant, but in 2018 all of the percentages more closely

matched the 12-year averages with clay still being slightly dominant (Table 10). In 2019, an anomaly occurs in AMT-

S data; an unexpectedly high portion of sand appears in the second and third of the three samples collected as the

first replicate shows the normal fine sand portion with a dramatic increase in silt (Figure 34). We suspect the second

and third PGS replicates, taken by a new sampling technician on the project, reflect digging deeper into the grab

sample than normal, i.e., rather than just the surface layer. Note that the PGS sample is always taken after the

chemistry sample and would not affect the chemistry assessment.

In 2020, all PGS sample parameters returned to their respective station’s centroids (Table 10).

4-12 Attachment A

2020 LTEMP Report

77

Figure 33. Individual replicate cumulative grain-size curves (%) for GOC-S and AMT-S, 2006-2020. 2020’s reps are

indicated by dotted red lines.

0

20

40

60

80

100

120

Clay Silt Sand

Cu

mu

lati

ve p

erc

en

t

GOC-S 2006-2020

0

20

40

60

80

100

120

Clay Silt Sand

Cum

ulat

ive

perc

ent

AMT-S 2006-2020

4-12 Attachment A

2020 LTEMP Report

78

Table 10. Average grain size components for GOC-S and AMT-S, 2006-2020.

AMT-S GOC-S

Year % Sand % Silt % Clay Year % Sand % Silt % Clay

2006 1 55 43 2006 6 44 51

20071 2 54 44 20071 7 37 56

2008 2 54 44 2008 9 36 55

20091 4 40 56 20091 10 35 55

2010 3 49 48 2010 9 38 53

2011 3 45 51 2011 16 32 52

2012 7 40 53 2012 17 27 56

2013 5 42 53 2013 7 37 56

2015 6 54 40 2015 8 64 28

2016 3 55 42 2016 4 56 40

2017 1 42 57 2017 2 49 50

2018 3 44 53 2018 2 52 46

2019 162 46 38 2019 8 42 49

2020 3 53 44 2020 2 50 48

avg 4 48 48 avg 8 43 50 1Combines two seasons of sampling 2Likely biased high

4-12 Attachment A

2020 LTEMP Report

79

Figure 34. 3D plots of grain size components from GOC-S and AMT-S 2006-2020. Sampling years are color coded:

blue to orange, 2006-2019; Red, 2020. Note the clipped axes emphasize a decreasing shift in clay vs silt at GOC-S in

2015. The two 2019 AMT-S outlier samples with anomalously high sand content (~20%) appear to be sampling

method errors.

AMT-S Grain Size

Replicates 2006-2020

2019

GOC-S Grain Size

Replicates 2006-2020

2020

2020

4-12 Attachment A

2020 LTEMP Report

80

APPENDIX 7. SEDIMENT TOC Total organic carbon (TOC as the percentage of sediment dry weight) serves as a non-specific measurement of all

organics in a sample. Typically ranging from 0.1 to 30% in marine sediments, it is used to express the nutritional

quality of food available to benthic organisms. For pollution work, metals and anthropogenic organic compounds

tend to sorb and concentrate in or on finer grained sediments and TOC, respectively, and thus TOC can be used to

normalize contaminant concentrations to do site-to-site contaminant comparisons.

During the more frequent samplings early in the LTEMP program, TOC values showed seasonal cycles (reflecting

spring plankton blooms) and with a slow increase in concentration plateauing around 2003 (Figure 35 and Table 11).

There was an uptick at GOC-S in 2012 and 2013 and mildly so at AMT-S in 2013. Since then, GOC-S TOC levels have

dropped in 2015-2019 and risen again in 2020, while AMT-S has ranged between 0.53 and 0.63% since 2012. Perhaps

the only conclusion is to note that TOC tends to fall within the low 0.5 – 0.8% DW range and suggests similar organic-

sparse sources at both locations within the fjord. This is not surprising considering the dominance of primarily

inorganic glacial flour in the sediments. For comparison, Port Valdez sediments collected for Alyeska’s Environmental

Monitoring Program (EMP, Shaw & Blanchard 2018, 2019) ranged between 0.3-0.6% (Figure 36). Intertidal sediments

collected in Cook Inlet ranged from 0.03 to 0.98% (Lees et al., 2001).

Also, note that the data are not continuous, that sampling prior to 2002 was performed by KLI, and that 2016 reflects

the third laboratory change for the project.

Figure 35. LTEMP Total Organic Carbon trends in AMT-S and GOC-S sediments (% DW), 1993-2020.

0.00

0.10

0.20

0.30

0.40

0.50

0.60

0.70

0.80

0.90

TOC

% d

ry w

t

AMT-S

GOC-S

4-12 Attachment A

2020 LTEMP Report

81

Table 11. LTEMP Total Organic Carbon in AMT-S and GOC-S sediments (% DW), 1993-2020.

Sample Date

AMT GOC Sample Date

AMT GOC Sample Date

AMT GOC

Apr-93 0.77 0.70 Mar-01 0.46 0.34 Jul-07 0.53 0.53

Jul-93 0.67 0.63 Jul-01 0.61 0.45 Apr-09 0.58 0.60

Mar-94 0.58 0.54 Mar-02 0.48 0.48 Jun-11 0.55 0.55

Jul-94 0.65 0.55 Jul-02 0.67 0.55 Jul-12 0.53 0.65

Apr-95 0.63 0.55 Mar-03 0.64 0.52 Jul-13 0.59 0.72

Jul-95 0.77 0.65 Jul-03 0.62 0.67 Jul-15 0.55 0.59

Mar-96 0.54 0.53 Mar-04 0.66 0.71 Jul-16 0.63 0.60

Jul-96 0.69 0.55 Jul-04 0.67 0.61 Jul-17 0.58 0.48

Mar-97 0.83 0.69 Mar-05 0.59 0.59 Jun-18 0.56 0.53

Jul-97 0.59 0.62 Jul-05 0.62 0.61 Jun-19 0.63 0.49

Mar-98 0.65 0.55 Mar-06 0.59 0.61 Jun-20 0.63 0.58

Apr-00 0.56 0.47 Jul-06 0.52 0.54

Jul-00 0.66 0.47 Apr-07 0.43 0.49

Figure 36. Total organic carbon for sediments in Alyeska’s 2017 and 2018 monitoring program (from Shaw &

Blanchard, 2018, 2019) in vicinity of the terminal (near shallow sites) and deeper within the fjord (far deep sites).

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

14

3A

143B

143C

14

5A

145B

145C

D2

5A

D25

B

D2

5C

D3

3A

D33

B

D3

3C

D5

1

D6

9 80

82

D7

3

D7

7 16

11

A

11B

11C 40

50

% T

OC

EMP TOC 2017 & 2018

2017

2018

Near Shallow Sites Far Deep Sites

4-12 Attachment A

2020 LTEMP Report

82

APPENDIX 8. OXYGENATED PRODUCTS IN TREATED BALLAST WATER DISCHARGES During the 2010 Deepwater Horizon event, a whole oil sample was analyzed on a high-resolution instrument, Fourier

transform-ion cyclotron resonance (FT-ICR; McKenna et al., 2013). While traditional oil spill forensics has relied upon

GC/MS and GC-FID instruments capably reporting ~300 hydrocarbons, the newer instrument saw an unexpected

universe of ~10,000 compounds. While it is unknown whether these are likely to be bioavailable or perhaps even

toxic, environmental monitoring based upon traditional chemical detection, to date, has “just been looking where

the light is good.”

As described in the 2017 report (Payne and Driskell 2018c), three effluent samples, raw, filtered (particulate oil

droplets), and dissolved phase, were collected from the BWTF discharge pipe in July 2016 and March 2017. In

addition to the standard PAH, SHC, and biomarkers analyzed as part of LTEMP, we independently (without PWSRCAC

support) had the July 2016 sample extracts screened for oxygenated products by a colleague, Dr. Christoph Aeppli

of Bigelow Laboratory (Maine). Oxygenated hydrocarbons, whether created microbially, by solar radiation, or by

chemical processes, are currently considered by hydrocarbon fate-and-weathering scientists to be the “Holy Grail”

in understanding oil-degradation products (Aeppli et al., 2012). Conceptually, the effluent from a ballast water

treatment facility designed to promote oil biodegradation would be an ideal substrate to use for oxidized-product

method development and validation.

To date, Dr. Aeppli has used an Iatroscan (TLC-FID) method to separate components in the July 2016 extracts into

saturated, aromatic, mono-oxygenates, and di-oxygenates. Extracts from fresh ANS crude oil and the three effluent

samples from July 2016 (Figure 37) showed the expected depletion of the saturate (SHC) and aromatic (PAH) in the

fresh ANS oil with their subsequent conversion into oxygenated products with one- and two-oxygen additions.

Because of the increased water solubility of oxygenated products, the highest relative concentrations of mono- and

di-oxygenated constituents were observed in the filtered, dissolved-phase fraction sample. Over 93% of the

measured components in that fraction were oxygenates compared to only 36% in the starting oil. These results

confirmed our expectations and help to document the biological treatment tank’s efficacy in converting

hydrocarbons into water-soluble, biodegradation-products. Subsequently, after discharge into the Port, oxygenated

products are more easily diluted and further weathered.

Continuing with method development, additional analyses are being undertaken using HPLC/MS, GC x GC/MS, and

by selected ion monitoring GC/MS after chemical derivatization into tri-methyl-silane (TMS) ethers and esters. To

date, a series of alcohols, carboxylic acids, diols, and dioic acids have been detected although explicit compound

identifications have not been completed. Nevertheless, these unfunded studies are expected to help expand this

line of investigation and may eventually help to track oxygenated products as they further degrade following

discharge from the BWTF or in future oil-spill releases.

In an approach that may be instructive to LTEMP projects, Sørensen et al., (2019) used very innovative chemical

analyses to characterize previously unquantified constituents of produced waters (PW) from five North Sea oil

platforms. Processing by fractionation and elution into polar and apolar fractions and derivatization allowed analysis

using gas chromatography (GC), GC-mass spectrometry (GC/MS), two dimensional GC/MS (GC × GC/MS) and liquid

chromatography with high-resolution spectrometry (LC-HRMS) techniques. A rich suite of polar and apolar

constituents were characterized and quantified within each fraction. Acute toxicity tests were then run using a

marine copepod subject. Toxicity varied significantly for different produced waters’ extractions and subfractions

with some different polar and apolar fractions being toxic within different produced water samples. Sørensen et al.

4-12 Attachment A

2020 LTEMP Report

83

Figure 37. Iatroscan (TLC-FID) of ANS oil, BWTF raw effluent, and BWTF dissolved components from July 2016 showing relative abundance of single- and double-

oxygenated (weathered) hydrocarbons relative to SHC and PAH components. Courtesy of Christoph Aeppli.

Raw effluent

Dissolved phase

ANS oil

SHC PAH Single-oxygen HC

Double-oxygen HC

4-12 Attachment A

2020 LTEMP Report

84

conclude, “Although, due to the vast chemical complexity even of the sub-fractions of the PW extracts, specific

compounds driving the observed toxicity could not be elucidated in this study, the proposed approach may suggest

a way forward for future revisions of monitoring regimes for PW discharges.” The BWTF extracts from previous (or

possibly future) LTEMP efforts may be ideal candidates for such analyses of biodegradation products as the extracts

may not contain the high levels of production chemicals (PCs) that were major constituents identified in the North

Sea produced waters.

APPENDIX 9. RELATED TOPICS These topics were presented in last year’s LTEMP report but have no additional updates this year. Being still relevant

to LTEMP monitoring, they are presented here for completeness.

Bioavailability of Particulate Phase Oil Hydrocarbons

Three studies should be mentioned regarding the assumption that only dissolved-phase hydrocarbons would be

bioavailable. In 2015, Auke Bay Lab (ABL) did a study for the PWSRCAC looking for oil in Port Valdez shrimp eggs

(Carls et al., 2016). They posited, as did others, that a clutch of eggs attached to the pleopods of gravid shrimp would

be exposed to and absorb a dissolved-PAH signal from residual hydrocarbons in the soft sediments. Although ABL

did not analyze for particulate-phase-confirming biomarkers, in at least one sample (Figure 38), the profile appears

to be a water-washed particulate profile. Either particulate oil is capable of infusing through a shrimp’s chitin eggshell

and inner lipid membranes or there was a problem with the lab’s sample cleanup methods, and a minor contribution

from particulate oil was present in the sample. Another study was more conclusive in showing a particulate signal in

eggs, this time on red crabs sampled on the abyssal plain in the Gulf of Mexico during the Deepwater Horizon

incident, which included a full suite of biomarkers (Douglas and Liu, 2015; G.S. Douglas, personal communication,

2015). Thus, crustacean eggs appear to absorb more than just dissolved-phase contaminants. If this is the case, then

the general supposition of dissolved-phase hydrocarbons’ exclusive bioavailability versus particulate-phase non-

bioavailability is perhaps over-simplified. This would certainly be the case when copepods that have ingested oil

micro-droplets in Port Valdez (Carls et al., 2006) are later consumed by predators (e.g., salmon smolt) or when

mussels filter micro-droplets from the water column.

Figure 38. PAH plot of shrimp eggs taken east of the VMT, which in our assessment, shows a water-washed,

weathered particulate-oil pattern absorbed through the chitin, lipid membrane and into the fat of the eggs. From

Carls et al., 2016.

4-12 Attachment A

2020 LTEMP Report

85

Low Level Toxicity Effects on Fish

Also notable is recent work documenting oil droplets unexpectedly adhering directly to certain fish species’ eggs

(but not others) (Laurel et al., 2019, Hansen et al., 2018, Sørenson et al., 2017, Sørhus et al., 2015) and demonstrating

increased toxicity relative to eggs with just dissolved component exposures. This work, so far, has just been looking

at three commercial fish species – cod, arctic cod, and haddock – with the latter one not susceptible to egg

membrane (chorion) oiling. Obviously, this research topic is in initial phases and far from garnering a deeper

understanding of the biomechanical differences in adsorbent versus non-adsorbent chorions. Therefore, it is

currently prudent to assume that all eggs in the water column, both vertebrate and invertebrate may be susceptible

to adhering oil droplets. Furthermore, this research is nascent; adhered or absorbed, insights into oils’ physiologic

and teratologic impacts on embryos are just developing. As concisely summarized by Laurel et al., (2019):

“Crude oil contains polycyclic aromatic hydrocarbons (PAHs) that are cardiotoxic. Three-ringed PAH families (e.g., phenanthrenes) enriched in crude oil block K+ and Ca2+ ion conductances in cardiomyocytes, disrupting the normal rhythmic pumping of the heart (Brette et al., 2014, 2017). When this occurs in oil exposed fish embryos, disruption of cardiac function leads to abnormal heart development (Incardona, 2017; Incardona and Scholz, 2016). Although cardiocirculatory defects alone would be sufficient to impact growth, more recent findings indicate that reduced cardiac function during embryonic and early larval development has other indirect effects that may be equally if not more consequential for individual fitness. Specifically, recent advances in RNA sequencing of oil-exposed Atlantic haddock (Melanogrammus aeglefinus) embryos identified alterations in the expression of genes involved in lipid metabolism (Sørhus et al., 2017). This suggests that disruption of bioenergetics during early development may be a prominent mechanism underlying latent impacts on fish growth and survival at later life stages. Oil spill science in marine systems has thus far focused on fish species with distinct ecophysiological characteristics (Incardona and Scholz, 2016). This includes nearshore and pelagic species spawning in cold northern waters (Carls et al., 1999; Incardona et al., 2015) and rapidly developing sub-tropical species (Incardona and Scholz, 2018). In general, cold water species or those with strong cold tolerance are more sensitive to oil-induced toxicity (Edmunds et al., 2015; Incardona et al., 2014, 2015; Morris et al., 2018; Sørensen et al., 2017; Sørhus et al., 2016). Although common morphological and functional abnormalities are usually evident shortly after embryonic exposure, delayed reductions in growth and juvenile survival have been documented in pink salmon exposed to low concentrations of oil that did not cause externally visible malformation (Heintz, 2007; Heintz et al., 2000). These effects on growth could reflect a latent and lasting dysregulation of lipid metabolism. If so, this would have important consequences for global marine fisheries because management paradigms are premised on a positive relationship between juvenile bioenergetics and successful recruitment to adult populations (Bouchard et al., 2017; Copeman et al., 2017; Heintz et al., 2013).”

Effects of Climate Change Enhanced Stream Flows on Sorptive Floc in the Water Column

In a more speculative vein, while the decline in environmental contaminants is obviously related to declining inputs

into the Port, another factor also seems relevant. Because discharged oil droplets sorb onto particulates in the water

column (Payne et al., 2003d and references therein), an increase in glacial flour brought into the Port from the nearby

Lowe and Valdez Glacier Rivers would affect dilution and settling rates of the oil. Streamflow records from the U.S.

Geographical Survey (USGS) from 2015-2019 do show high stream flows in the Valdez Glacier River but mostly

normal flows from Lowe River (Figure 39). We might speculate that with climate change accelerating the melt of

glaciers, it seems intuitive to expect higher flows and thus, greater flocculant loads delivered to the Port environs

and potentially increased dilution of oil signatures in the sediment. The degree of relevance to sediment

hydrocarbon loads measured by LTEMP and Alyeska is unknown.

4-12 Attachment A

2020 LTEMP Report

86

Figure 39. Historic monthly stream discharge rates (cubic ft/sec) for Lowe River and Valdez Glacier Stream, 2015-

2019 (from USGS 2019).

0

1000

2000

3000

4000

5000

6000

Jul-

14

Sep

-14

No

v-1

4

Jan-

15

Mar

-15

May

-15

Jul-

15

Sep

-15

No

v-1

5

Jan-

16

Mar

-16

May

-16

Jul-

16

Sep

-16

No

v-1

6

Jan-

17

Mar

-17

May

-17

Jul-

17

Sep

-17

No

v-1

7

Jan-

18

Mar

-18

May

-18

Jul-

18

Sep

-18

No

v-1

8

Jan-

19

Mar

-19

cu F

t/se

c

Monthly mean stream discharge

Lowe River

Valdez Glacier

4-12 Attachment A

2020 LTEMP Report

87

Seasonal Variability of TPAH Related to Lipid-reducing Spawning Events

Finally, recall three observations from previous sections, 1) there are substantial seasonal differences in the BWTF

effluent profiles and concentrations, 2) the mussels sampled at the Berth 5 spill in 2017 purged the oil in less than

three months, and 3) spiking volumes of freshwater inputs loaded with glacial flour occur during the summer

sampling period. A fourth point is that hydrocarbons tend to infuse into and accumulate in an organism’s lipids (e.g.,

eggs). Therefore, a spawning event may greatly reduce accumulated oil-contaminant loads in mussels if they’ve

spawned just prior to sampling. LTEMP sampling generally occurs in late June-early July, presumably post-spawning.

Together, these observations suggest that hydrocarbon contaminate loads likely vary substantially throughout the

year. While the limited LTEMP results suggest an exceptionally clean environment, it is a presumptuous conclusion

when little is known about off-schedule conditions. As such, we suggest a pilot project doing more frequent mussel

sampling be considered to gather more insights into current mussel contaminant variations.

APPENDIX 10. BEYOND LTEMP How do the measured tissue-hydrocarbon levels compare with other Alaska sites? Unfortunately, comparable

studies are scarce, no longer current, and variable concerning which analytes are actually summed. Nevertheless,

earlier reported values still seem reasonable (Table 12). Follow-up mussel sampling in 2004 for oil residues from the

1997 M/V Kuroshima grounding in Summer Bay, Unalaska, found TPAH levels between 25 and 85 ng/g DW, with an

average of 57 ng/g DW (Helton et al., 2004). This is actually higher than LTEMP’s July 2013 range of 9-28 ng/g DW

from the six stations inside the Sound but similar to the 13-65 ng/g DW range at three GOA sites (overall average 50

ng/g DW) collected at the same time. Compared to the 2018 data for all three LTEMP regions, the 2004 Kuroshima

sites were significantly higher, but there are no more recent data against which to compare. The current LTEMP data

suggest a natural dissolved-phase background TPAH somewhere between 33-71 ng/g DW.

Reaching farther, data from the 2004-2005 National Status and Trends, Mussel Watch Program (Figure 40) and 2008-

2010 Alaska sites (Figure 41 and Table 12) (now summing only 38 parent and alkylated PAH homologues versus 43

LTEMP PAH analytes) show that average TPAH concentrations in mussels for other West Coast sites have been nearly

12-25 times higher (825 ng/g DW) than LTEMP’s current levels. The highest level reported on the West Coast was

6,962 ng/g DW in Seattle, Washington. The lowest, 63 ng/g DW, was from mussels collected on Santa Catalina Island,

26 miles offshore of Orange County in Southern California. In 2004, the average TPAH mussel concentration in

mussels from the five Alaska Mussel Watch sites (Ketchikan, Nahku Bay, Port Valdez, Unakwik Inlet, and Cook Inlet)

was 267 ng/g DW with levels ranging from 105-441 ng/g DW (Kimbrough et al., 2008). In 2010, the average from

four Alaska sites (Nanwalek, Port Gram (two stations), and Seldovia Bay) was 413 ng/g DW while Nash Road in

Resurrection Bay exceeded all the other stations at 1,570 ng/g DW. Considering these and even more recent 2008-

2010 values from the Mussel Watch data portal, the LTEMP results demonstrate that these remote locations are still

exceptionally clean.

Finally, a 2005 EVOS Trustee Council Program, Long-Term Monitoring of Anthropogenic Hydrocarbons in the Exxon

Valdez Oil Spill Region, examined 10 intertidal sites within the Naked-Knight-Southwest Island complex to measure

the extent of buried oil still present 16 years after the spill. At previously heavily oiled EVOS sites, 10 to 50 random

pits (depending on the beach width) were excavated to a depth of ~0.5 m to look for residual oil. If oil was detected,

available nearby mussels were also collected. The results have been published elsewhere (Short et al., 2007) but, as

co-authors, PAH and SHC sample profiles were included in Appendix E of our 2005/2006 Report (Payne et al., 2008a).

Briefly, TPAH levels in the oiled pits ranged from a low of 42 ng/g (on Knight Island) to a high of 567,000 ng/g (on

4-12 Attachment A

2020 LTEMP Report

88

Table 12. Most recent TPAH concentrations in LTEMP mussel tissues (ppb, ng/g DW) relative to 2004-2010 NOAA

Mussel Watch monitoring data and a recovered Alaska oil spill event.

LTEMP 2020 2019

Port Valdez tissues Port Valdez tissues

AMT-B, JAP & GOC AMT-B, JAP & GOC

33-71 24-34

2018 Port Valdez tissues AMT-B, JAP & GOC 29-40 2017 Port Valdez tissues AMT-B, JAP & GOC 46-63 2016 Port Valdez tissues AMT-B, JAP & GOC 69-195

2015 Port Valdez tissues AMT-B & GOC 39-87 2013 Port Valdez tissues AMT-B & GOC 17-20

2018 PWS Five stations 22-38 GOA Three stations 21-29

2015 PWS (KNH & SHB) KNH & SHB 15-27 2013 PWS Six stations 9-28

GOA Three stations 13-65

West Coast Mussel Watch

average (Kimbrough et al., 2008) 825

So. Calif. Santa Catalina Island 63 Seattle Elliot Bay, WA 6,962

Alaska Mussel Watch 2010

Resurrection Bay Nash Road 1,570

Nanwalek Nanwalek 194 Port Graham Port Graham 376 Port Graham Murphy's Slough 428 Seldovia Bay Powder Island 652

2009 Kachemak Bay Chrome Bay 173 Kachemak Bay Tutka Bay 485

Ketchikan Mountain Point 231 Nahku Bay East Side 229 Port Valdez Mineral Creek Flats 332 Resurrection Bay Nash Road 602 Resurrection Bay Nash Road 765 Resurrection Bay Nash Road 929 Resurrection Bay Nash Road 713 Unakwit Inlet Siwash Bay 257

2008 Cook Inlet Bear Cove 119 Cook Inlet Homer Spit 208 Port Valdez Alyeska Marine Terminal 52 Port Valdez Gold Creek 31

M/V Kuroshima (1997) 2004

Unalaska

25-85

Latouche Island) with the oil showing states of weathering varying from extensively degraded to very fresh. On the

other hand, nearby mussel samples only showed low dissolved-phase TPAH concentrations (11-42 ng/g DW, derived

primarily from naphthalenes and phenanthrenes/anthracenes) that were in the same concentration range but

compositionally different from the signals observed at LTEMP PWS and GOA sites sampled in April 2009. From these

studies, it was concluded that although in 2005, there were still persistent buried EVOS residues at a number of

beaches, they were highly sequestered and did not appear to be bioavailable unless disturbed.

4-12 Attachment A

2020 LTEMP Report

89

Figure 40. Status and trends result from National Mussel Watch data (Kimbrough et al., 2008). All Alaska sites characterized as low concentrations.

4-12 Attachment A

2020 LTEMP Report

90

Figure 41. Summary page of Alaska regional Mussel Watch results and trends based on 2004-05 report from Kimbrough et al.,

2008.

The low 2018 TPAH and 2019 values in mussels collected from these sites and the PSD data from the formerly oiled

DII site vs. the clean-control site at KNH (Figure 25) seem to confirm these findings, although recent studies with oil-

sniffing dogs suggest that bioavailability at concentrations below our analytical detection limits may still be a concern

for sub-lethal effects with some species. In 2005, rates of EVOS oil disappearance had diminished to an estimated

4% per year. If left undisturbed, Short et al., (2007) predicted that sequestered hydrocarbons would be there for

decades. Revisiting the sites in 2015, Auke Bay researchers found mostly unchanged conditions since 2001

(Lindeberg et al., 2018). Lindeberg again concluded that an estimated 0.6% EVOS oil would remain sequestered

unless disturbed and will likely persist in the environment on a decadal scale. They also comment that viewing this

survey in the context of previous surveys makes it clear that Exxon researcher claims made after the spill that

beaches would clean themselves were overly optimistic. To address these residual deposits, the EVOS Trustee

Council has sponsored various beach remediation studies and pilot projects (ADEC 2016).

4-12 Attachment A

Using Mussel Transcriptomics for Environmental Monitoring in Port Valdez,

Alaska: 2019 and 2020 Pilot Study Results

PWSRCAC Contract number: 951.21.06

February 17, 2021

Lizabeth Bowen1, Austin Love2, Shannon Waters1, Katrina Counihan3, Brenda Ballachey4,

Heather Coletti5, William B. Driskell6, and James R. Payne7

1U.S. Geological Survey, Western Ecological Research Center, Davis, CA 95616 USA.

[email protected]. 530-752-5365

2Prince William Sound Regional Citizens’ Advisory Council, Valdez, AK, USA.

3Alaska SeaLife Center, Seward, AK 99664 USA

4U.S. Geological Survey (Emeritus), Alaska Science Center, Anchorage, AK, USA

5National Park Service, Inventory & Monitoring Program, Southwest Alaska Network,

Anchorage, AK, USA

6Independent Consultant, Seattle, WA, USA

7Payne Environmental Consultants, Inc., Encinitas, CA, USA

The opinions expressed in this PWSRCAC-commissioned report are not necessarily those

of PWSRCAC.

4-12 Attachment B

mailto:[email protected]

1

TABLE OF CONTENTS

ACRONYMS .................................................................................................................................... 2

INTRODUCTION ............................................................................................................................ 4

METHODS ....................................................................................................................................... 6

RESULTS ....................................................................................................................................... 15

CONCLUSIONS............................................................................................................................. 39

RECOMMENDATIONS ................................................................................................................ 40

REFERENCES ............................................................................................................................... 43

APPENDIX ..................................................................................................................................... 47

4-12 Attachment B

2

ACRONYMS

AMT – Saw Island sample site

ANS – Alaska North Slope

B[a]P – benzo[a]pyrene

CaM – Calmodulin gene

Casp8 – Caspase 8 gene

CCOIV – Cytochrome C Oxidase IV gene

cDNA – complementary deoxyribonucleic acid

CHI – Chitinase gene

CNN – Calponin gene

Cyp3 – Cytochrome P450 family 3 gene

CT – cycle threshold crossing values

GB – Galena Bay sample site

GoC – Gold Creek sample site

Harbor – Whittier Harbor, Seward Harbor and Cordova Harbor sample sites

HIFa – hypoxia-inducible factor alpha gene

HSP70 – heat shock protein 70 gene

HSP90 – heat shock protein 90 gene

JPO – Jackson Point sample site

JB – Jack Bay sample site

KATM – Katmai National Park

LACL – Lake Clark National Park

LTEMP – Long-Term Environmental Monitoring Program

mRNA – messenger ribonucleic acid

MIF – macrophage migration inhibitory factor gene

MT20 – Metallothionein 20 gene

Myt – Mytilin gene

MytB – Myticin B gene

NMDS – non-parametric multidimensional scaling

NPP- National Park and Preserve

PAH – polycyclic aromatic hydrocarbons

PCR – polymerase chain reaction

PWS – Prince William Sound

PWS Field – Herring Bay, Hogan Bay, Iktua Bay, Johnson Bay, and Whale Bay sample sites

PWSRCAC – Prince William Sound Regional Citizens’ Advisory Council

P53 – tumor protein 53 gene

4-12 Attachment B

3

RNA – ribonucleic acid

SAC – Scientific Advisory Committee

USGS – United States Geological Survey

VH – Valdez Small Boat Harbor sample site

18S – ribosomal reference gene

4-12 Attachment B

4

INTRODUCTION

This project was inspired by a Prince William Sound Regional Citizens’ Advisory Council

(PWSRCAC) 2018 Science Night presentation of work by Dr. Katrina Counihan from the Alaska

SeaLife Center. The results comprised mussel and razor clam genetic transcription experiments she

conducted in Southcentral Alaska. Through Dr. Counihan’s work, the Council was introduced to

another researcher, Dr. Lizabeth Bowen, an ecologist with the United States Geological Survey

(USGS), who was the lead author of a 2018 research paper titled “Gene transcription patterns in

response to low level petroleum contaminants in Mytilus trossulus [Bay mussels] from field sites

and harbors in southcentral Alaska,” in Deep-Sea Research Part II.

Dr. Bowen’s 2018 paper looked at whether mussel gene transcription is affected by oil

pollution and other environmental factors. Dr. Bowen’s research included five locations in Prince

William Sound (PWS), Alaska, that were oiled during the Exxon Valdez oil spill and three harbors

in the Exxon Valdez oil spill region: Seward, Whittier, and Cordova.

Exposure to contaminants, including polycyclic aromatic hydrocarbons (PAHs) found in

crude oil, can lead to pathophysiological changes that may be subtle but significant, and difficult to

detect using classical biological or chemical diagnostic methods. The earliest observable

indications of physiological impairment can be altered levels of gene transcripts, evident prior to

clinical signs (Farr and Dunn, 1999; McLoughlin et al., 2006; Poynton and Vulpe, 2009). Gene

transcription is the process by which information from the DNA template of a particular gene is

transcribed into messenger ribonucleic acid (mRNA) and eventually translated into a functional

protein. Quantitative analysis of mRNA therefore is used as a measure of gene transcription (Heid

et al., 1996). The amount of a particular gene that is transcribed is physiologically dictated by a

number of intrinsic and extrinsic factors, including stimuli such as infectious agents, toxin

exposure, trauma, or neoplasia. For the interests of this study, gene transcription assays measure the

4-12 Attachment B

5

physiological response of an organism to xenobiotic oil contaminants. Furthermore, the deleterious

effects of toxic exposure may persist beyond metabolism and excretion of the toxin. Gene

expression assays are advantaged by the ability to measure the persistent physiologic responses of

an individual to the metabolic stimuli, independent of the continued presence of the original toxin

or its metabolites.

This two-year project is a pilot study designed to mirror previous mussel transcriptomic

research conducted by Dr. Bowen and Dr. Counihan in Southcentral Alaska. The goal is to

determine if transcriptomic analysis of mussel tissue would be useful as a part of the Council’s

Long-Term Environmental Monitoring Program (LTEMP).

4-12 Attachment B

6

METHODS

Mussel collection and tissue preservation

Mussels were collected in June 2019 and 2020, from each of four locations in Port Valdez:

Saw Island (AMT), Jackson Point (JPO), Gold Creek (GoC), and the Valdez Small Boat Harbor

(VH) (Figure 1). AMT, JPO, and GoC are “traditional” LTEMP sites. VH, a location of known

mixed petroleum and pyrogenic contamination, was sampled as a strong “positive control” site to

better understand how transcription reacts to chronic levels of petroleum contamination. In 2019,

the VH mussels were collected from a creosote piling. We note that creosote leachates are not the

same weathered Alaska North Slope (ANS) crude oil being released at the terminal albeit similar

constituents are present. In 2020, the VH mussels were collected from boulder riprap exposed to

vessel traffic in the harbor entrance where, although still not weathered ANS crude oil, there are

sources of ANS derived fuel and oils in the chemical profiles.

In contrast to VH, GoC is relatively far from known sources of hydrocarbon contamination;

therefore, it was chosen as the clean control site to compare to other sampling locations. However,

the June 2019 transcriptomics results indicated that GoC may not be as free from petroleum

contamination as hoped. In June 2020, two additional sites (Jack Bay (JB) and Galena Bay (GB))

more remote in Valdez Arm were sampled as alternative clean control sites (Figure 2).

The GB location was an intertidal site previously sampled through the Gulf Watch Alaska’s

environmental monitoring program – but is currently not actively sampled in that program. The JB

site is not known to have been previously sampled by the Gulf Watch or other environmental

monitoring programs but was selected due to its distance from known sources of petroleum

contamination.

Procedurally, after the mussels were collected their gill and adductor muscle tissues were

extracted, preserved in RNAlater, and frozen for further analysis. At each sampling location

4-12 Attachment B

7

mussels were sampled from various heights of the intertidal zone. Three replicate samples of 10

mussels were collected from each location. The 10 mussels for each replicate were generally

collected along a 2 meter section of shoreline. Relatively larger mussels were sampled for ease of

dissecting gill and adductor tissues, and no morphometric data was collected.

On April 12, 2020, there was an oil spill from the Valdez Marine Terminal. Additional

mussels were collected from April through August 2020, for transcriptomic and chemical

analysis. The results of those analyses associated with the mussels collected in response to the

April 12 oil spill will be described in a separate report and manuscript.

4-12 Attachment B

8

Figure 1. Map of Port Valdez showing the four mussel transcriptomics locations sampled in

2019 & 2020 (Source: Google Earth).

Figure 2. Map of the Valdez Arm showing the two mussel transcriptomics locations

sampled in 2020 (Source: Google Earth).

4-12 Attachment B

9

Previously Available Samples

Although this study was focused on sites in Port Valdez and the Valdez Arm (LTEMP

sites), we had the opportunity to compare these samples with previously published data (Bowen et

al. 2018, Counihan et al. 2019). These data include mussel samples from Katmai (KATM) and

Lake Clark National Parks (LACL) in 2015-2016 (Figure 3), as well as various locations in and

around PWS in 2012-2015 (Figure 4). The 2012-2015 PWS samples include mussels taken from

three harbor locations (Whittier, Seward, and Cordova Harbor) and five field locations (Herring

Bay, Hogan Bay, Iktua Bay, Johnson Bay, and Whale Bay). The PWS samples included both gill

and adductor tissue, whereas the KATM and LACL samples only included gill tissue (Table 1).

During the 2020 LTEMP sampling event, 69 samples were collected and analyzed (Table 1). The

transcription of the 15 genes was analyzed in all 69 samples (Table 2).

Figure 3. Mussel sampling sites in Lake Clark National Park and Preserve (NPP) and

Katmai NPP. Sites were sampled during June of 2015 and 2016.

4-12 Attachment B

10

Figure 4. Location of harbor and field mussel sampling sites in Prince William Sound.

Table 1. Numbers of samples of gill and adductor muscle tissues sampled by site. KATM and

LACL each include three separate sites. PWS includes five field sites and three harbor sites.

Abbreviation Gill Adductor Location & Collection Year

AMT19 10 10 Saw Island 2019

AMT20 10 10 Saw Island 2020

GB20 10 10 Galena Bay 2020

GoC19 10 10 Gold Creek 2019

GoC20 10 10 Gold Creek 2020

JB20 10 10 Jack Bay 2020

JPO19 10 10 Jackson Point 2019

JPO20 10 10 Jackson Point 2020

KATM 60 0 Katmai National Park 2015-2016

LACL 60 0 Lake Clark National Park 2015-2016

PWS 23 90 Prince William Sound 2012-2015

VH19 10 10 Valdez Small Boat Harbor 2019

VH20 19 19 Valdez Small Boat Harbor 2020

4-12 Attachment B

11

Table 2. Genes selected for the transcription panel, the primary biological processes they are

associated with, and what types of environmental interactions are known to affect their

transcription.

Gene Biological Process Environmental

Interaction References

Calmodulin (CaM) Metabolism, shell

formation

Ocean acidification

Temperature Dissolved oxygen

Chen et al. (2012);

Li et al. (2004)

Caspase 8 (Casp8) Programmed cell death, necrosis, inflammation

Pathogens Contaminants

Romero et al. (2011)

Macrophage

migration

inhibitory factor

(MIF)

Innate immunity

Pathogens

Parisi et al. (2012),

Philipp et al. (2012)

Calponin (CNN) Hypoxia Ocean acidification Dissolved oxygen

Hüning et al. (2013),

Li et al. (2016)

Chitinase (CHI) Metabolism, hypoxia Ocean acidification Dissolved oxygen

Banni et al. (2011),

Hüning et al. (2013)

Cytochrome C Oxidase IV

(CCOIV)

Hypoxia

Dissolved oxygen Fukuda et al. (2007)

Heat shock protein

70 (HSP70)

Thermal stress

Temperature, Pathogens, Contaminants

De Maio (1999),

Iwama et al. (1999),

Tsan & Gao

(2004)

Heat shock protein

90 (HSP90)

Thermal stress Temperature Pathogens

Contaminants

De Maio (1999),

Iwama et al. (1999),

Tsan & Gao

(2004)

Hypoxia-inducible factor alpha (HIFa)

Hypoxia Dissolved oxygen Wu (2002)

Myticin B (MytB) Innate immunity Pathogens Balseiro et al. (2011)

Mytilin (Myt) Innate immunity Pathogens

Ocean acidification Balseiro et al. (2011),

Mitta et al. (2000)

Metallothionein 20 (MT20)

Detoxification Contaminants -

metals Banni et al. (2007)

Cytochrome P450, family 3 (Cyp3)

Detoxification Contaminants Giuliani et al.

(2013)

Tumor protein 53 (P53)

Programmed cell

death

Contaminants - PAHs

Goodson et al.

(2006),

Banni et al. (2009)

Ribosomal 18S

(18S)

Method reference

gene

Low interaction

potential Counihan et al.

2019

4-12 Attachment B

http://en.wikipedia.org/wiki/Necrosis

http://en.wikipedia.org/wiki/Inflammation

12

RNA extraction

Total RNA was extracted from homogenized adductor muscle and gill tissue using the

RNeasy Lipid Tissue Mini Kit (Qiagen, Valencia, CA). To remove contaminating genomic

DNA, the spin columns were treated with 10 U μL−1 of RNase-free DNase I (DNase, Amersham

Pharmacia Biotech Inc.; www.apbiotech.com) at 20 °C for 15 minutes (min) followed by

extraction of total RNA and stored at −80 °C.

cDNA synthesis

A standard complementary deoxyribonucleic acid (cDNA) synthesis was performed on 2

μg of RNA template from each tissue. Reaction conditions included 4 units reverse transcriptase

(Omniscript, Qiagen, Valencia, CA), 1 μM random hexamers, 0.5 mM each dNTP, and 10 units

RNase inhibitor, in RT buffer (Qiagen, Valencia, CA). Reactions were incubated for 60 min at

37 °C, followed by an enzyme inactivation step of 5 min at 93 °C, and then stored at –30 °C until

further analysis. Real-time polymerase chain reaction (PCR) reactions for the individual, mussel-

specific housekeeping gene (18S) and genes of interest were run in separate wells (Table 2).

Briefly, 1 µl of cDNA was added to a mix containing 12.5 μl of Applied Biosystems Fast

SYBR Green® Master Mix [5 mM Mg2+] (Qiagen, Valencia, CA), 0.5 μl each of forward and

reverse sequence specific primers (Invitrogen, Carlsbad, CA), and 10.5 μl of RNase-free water;

total reaction mixture was 25 μl. The reaction mixture cDNA samples for each gene of interest

and 18S were loaded into Fast SYBR Green® 96 well plates in duplicate and sealed with optical

sealing tape (Applied Biosystems, Foster City, CA). Reaction mixtures that contained water but

no cDNA were used as negative controls. Amplifications were conducted on a Step-One Plus

Real-time Thermal Cycler (Applied Biosystems, Foster City, CA). Reaction conditions were as

follows: 50 °C for 2 min, 95 °C for 15 min, 40 cycles of 94 °C for 30 seconds (s), 60 °C for 30 s,

4-12 Attachment B

13

72 °C for 31 s, and an extended elongation phase at 72 °C for 10 min. Reaction specificity was

monitored by melting curve analysis using a final data acquisition phase of 60 cycles of 65 °C for

30 s and verified by direct sequencing of randomly selected amplicons. Cycle threshold crossing

values (CT) for the genes of interest were normalized to the 18S housekeeping gene. The CT

value of a reaction is defined as the PCR cycle number when the fluorescence of a PCR product

can be detected above the background signal and is associated with the amount of PCR product in

the reaction. Thus, the lower the CT value, the more PCR product that is present.

Statistical analysis

Separate statistical analyses were conducted for the two tissue types, adductor muscle and

gill. Transcriptomic responses to stress are likely related to the specific physiological role of

each tissue in the organism (Kadota et al. 2003) and thus, generally tissue specific. Analysis of

quantitative PCR data was conducted using normalized values (ribosomal housekeeping gene

threshold crossing subtracted from the gene of interest’s threshold crossing). Note that while

lower normalized values are indicative of higher numbers of transcripts, we have inverted the

values in the box plots for ease of interpretation. A change in normalized value of two is

approximately equivalent to a four-fold change in the amount of the transcript. The measured

gene expression variation between samples is the sum of the true biological variation and

several confounding factors (e.g., method limit differences in pipetting volume or sample

adhesion to plastic wells, etc.) resulting in non-specific variation. The goal of normalization is

to remove the non-biological variation as much as possible.

For descriptive analysis, median transcript values were calculated for each tissue at each

location (Table 3 and Table 4 in APPENDIX). Tissues were analyzed individually, not as

composites, and data are generally represented by site medians. Medians were used to avoid the

4-12 Attachment B

14

effects of outliers, which are kept in the data as they convey important information. Correlations

between genes were calculated using a Pearson correlation matrix with heat map visualization

(NCSS© Statistical Software, 2007, Kaysville, Utah).

Conventional median responses per group (based on location) were assessed for

statistical significance between classification ranks using Kruskal-Wallis with Dunns’ Multiple

Comparison Tests and Bonferroni correction, with reported Z values (NCSS© Statistical

Software, 2007, Kaysville, Utah). The 2019 and 2020 LTEMP sites mussel adductor and gill

tissue samples were compared separately by location. Another analysis grouped the 2019

LTEMP adductor muscle samples and the 2020 LTEMP adductor muscle samples to then

compare them with the median of the PWS adductor samples. The 2019 and 2020 LTEMP gill

tissue samples were compared with mussel gill samples collected in 2015 and 2016 at KATM

and LACL, as well as with samples collected in 2012-2015 in PWS.

We constructed three-dimensional scatter plots to visualize transcript levels by the

various genes, tissue type, location, and year (Miner 3D ENTERPRISE, Paris, France). The 3-D

graphical representations show median transcript levels by location clustered by similarity in

transcription and not by pre-defined groups. Similar transcript levels group closer together in

three-dimensional space.

4-12 Attachment B

15

RESULTS

Median responses

Median CT values for genes of interest for mussel adductor and gill tissues were calculated

based on location and year (Table 3 and Table 4 in APPENDIX). Note that smaller values

indicate greater levels of transcription. For comparison, median responses are depicted from

mussels sampled at the complied PWS sites (field and harbors), as well as KATM and LACL

(Bowen et al. 2018, Counihan et al. 2019).

Correlations

The Pearson correlation coefficient, r, can take a range of values from +1 to -1 where a

value of 0 indicates that there is no association between the two variables and a value closer to

+1 or -1 indicates a stronger association. Values greater than 0 indicate a positive association (as

the value of one variable increases, so does the value of the other variable), while values less

than 0 indicate a negative association (as the value of one variable increases, the value of the

other variable decreases). Heat maps were generated for the correlation matrices for adductor

and gill (Figure 5 and Figure 6). Note that the rows and columns are sorted in the order

suggested by hierarchical clustering. This plot graphically associates various “like” subsets of

the variables that seem to be highly correlated within the subset. For example, adductor muscle

tissues show no negative (blue) correlations, while for gill tissue, there are slight negative (light

blue) correlations.

4-12 Attachment B

16

Figure 5. Heat map of LTEMP adductor muscle tissue correlation matrix

4-12 Attachment B

17

Figure 6. Heat map of LTEMP gill tissue correlation matrix

Considerations for adductor and gill tissue results

Transcriptional patterns are generally tissue specific. These tissue-specific responses are

likely related to the specific physiological role of each tissue in the mussel, a phenomenon that

has been found in many studies. Additionally, it has been hypothesized that the specialized

functions of different tissues could make some tissues more or less susceptible to disruptions

4-12 Attachment B

18

from contaminant exposure. Thus, we have chosen to examine both gill and adductor muscle

tissues for breadth of understanding.

For both adductor muscle and gill tissues, the next analyses were focused on the three

genes most directly linked with response to hydrocarbon exposures, MT20, Cyp3, and P53. For

both adductor and gill tissue, gene transcript levels associated with contaminant presence that

were significantly different among locations included MT20, Cyp3, and P53. Although the

remainder of the genes in our panel are not generally associated with detoxification, many are

influenced by the presence of contaminants. For example, recent research by Banni et al. 2017,

shows that controlled exposure to B[a]P results in transcriptional changes of CNN, CaM, Myt,

CCOIV, and CHI. These are only a few of the indirect or “downstream” effects of contaminant

exposure. The mechanism is as yet unclear, but an initiation of the detoxification pathway in this

case results in effects on shell formation, mitochondrial activity, and immune function (Banni et

al. 2017). In general, the 2019 and 2020 LTEMP samples, from Port Valdez and the Valdez

Arm, had higher levels of transcription in genes associated with detoxification, MT20, Cyp3, and

P53, than KATM, LACL, and PWS samples.

Adductor muscle tissue

The 2019 and 2020 transcript profiles of adductor tissue from Port Valdez and Valdez Arm

mussels (LTEMP sites) were compared with transcript profiles from 2012 – 2015 PWS mussels.

Gene transcript levels associated with contaminant presence that were significantly different

among locations included MT20, Cyp3, and P53 (Figures 7-12). Figure 7, Figure 8, and Figure

9 depict genes in mussel adductor tissue primarily associated with detoxification of contaminants

including PAHs, compared among individual LTEMP sites (AMT, GB, GoC, JB, JPO, and VH).

When sampled twice, sites were split into the 2019 and 2020 sampling years. These genes were

4-12 Attachment B

19

also compared among broader groupings of sites (LTEMP 2019, LTEMP 2020, and PWS; Figure

10, Figure 11, and Figure 12). The following box-and-whisker quartile plots use bars to depict

the range from the 10th to the 90th percentile of individual normalized transcription values for

each gene. The rectangular box indicates the 1st and 3rd quartiles around the median mid-line. Red

circles, if present, represent 5th and 95th percentile outliers. Interpretation of sampling location

and gene abbreviations are provided in Table 1 and Table 2.

4-12 Attachment B

20

Figure 7. MT20 transcript levels in adductor muscle tissues collected as part of 2019 and

2020 LTEMP.

For MT20, indicative of hydrocarbon as well as metal exposure, the highest transcript levels in

adductor muscle tissue were found in mussels sampled in VH19. Data were assessed for

statistical significance between locations/years using Kruskal-Wallis with Dunns’ Multiple


Software, 2007, Kaysville, Utah). “>” can be translated as “has higher transcription than.”

Significant differences: GoC19 > AMT19, VH20; VH19 > AMT19, AMT20, VH20 (P = 0.00).

4-12 Attachment B

21

Figure 8. Cyp3 transcript levels in adductor muscle tissues collected as part of 2019 and 2020

LTEMP.

For Cyp3, indicative of contaminant detoxification activities including PAHs, the highest transcript

levels in adductor muscle tissue were found in mussels sampled at VH19. Data were assessed for


Comparison Tests and Bonferroni correction, with reported Z values (NCSS© Statistical Software,

2007, Kaysville, Utah). “>” can be translated as “has higher transcription than.” Significant

differences: VH19 > AMT19, GB20, GoC20, JPO19 (P = 0.00)

4-12 Attachment B

22

Figure 9. P53 transcript levels in adductor muscle tissues collected as part of 2019 and 2020

LTEMP.

For P53, primarily indicative of cell death and malignant transformation, as well as PAH

exposure, the highest transcript levels in adductor muscle tissue were found in mussels

sampled in VH19. Data were assessed for statistical significance between locations/years

using Kruskal-Wallis with Dunns’ Multiple Comparison Tests and Bonferroni correction, with

reported Z values (NCSS© Statistical Software, 2007, Kaysville, Utah). “>” can be translated

as “has higher transcription than.” Significant differences: VH19 > AMT19, AMT20, GB20,

GoC20, JB20, VH20 (P = 0.00).

4-12 Attachment B

23

Figure 10. MT20 transcript levels in adductor muscle tissues collected as part of LTEMP

2019 and 2020, and Prince William Sound.


adductor muscle tissue were found in mussels sampled at the LTEMP 2019 and 2020 sites in

comparison with sites in PWS (Field and Harbors). Data were assessed for statistical

significance between locations/years using Kruskal-Wallis with Dunns’ Multiple Comparison

Tests and Bonferroni correction, with reported Z values (NCSS© Statistical Software, 2007,

Kaysville, Utah). “>” can be translated as “has higher transcription than.” Significant

differences: LTEMP19 and LTEMP20 > PWS (P=0.00).

4-12 Attachment B

24

Figure 11. Cyp3 transcript levels in adductor muscle tissues collected as part of LTEMP 2019

and 2020, and PWS.

For Cyp3, indicative of contaminant detoxification activities including PAHs, the highest

transcript levels in adductor muscle tissue were found in mussels sampled at the LTEMP 2019

and 2020 sites in comparison with sites in PWS (including harbors). Data was assessed for




Significant differences: LTEMP19 and LTEMP20 > PWS (P=0.00).

4-12 Attachment B

25

Figure 12. P53 transcript levels in adductor muscle tissues collected as part of LTEMP 2019

and 2020, and PWS.

For P53, primarily indicative of cell death and malignant transformation, as well as PAH exposure,

the highest transcript levels in adductor muscle tissue were found in mussels sampled at the

LTEMP 2019 and 2020 sites in comparison with sites in PWS (including harbors). Data was

assessed for statistical significance between locations/years using Kruskal-Wallis with Dunns’

Multiple Comparison Tests and Bonferroni correction, with reported Z values (NCSS© Statistical


Significant differences: LTEMP19 and LTEMP20 > PWS (P=0.00).

4-12 Attachment B

26

Figure 13 and Figure 14 depict three-dimensional scatter plots of the median values for

MT20, Cyp3, and P53, in adductor tissue, at the 2019 and 2020 LTEMP sites (VH, JPO, JB,

GoC, GB, and AMT) compared to the 2012-2015 PWS locations. Figure 13 and Figure 14 only

differ in the angle of their orientation. Our results show that the highest levels of transcription of

the three genes are found in mussels sampled at the VH in 2019 (black cube closest to axes

origin). These samples were taken from mussels attached to a creosote piling and thus served as

a strong reference for contaminant exposure. The lowest levels of transcription for each of the

genes is found in mussels sampled in PWS (blue pyramid), while the remaining LTEMP

samples fell in between VH19 and PWS.

4-12 Attachment B

27

Figure 13. Three-dimensional scatter plots of the median values for MT20, Cyp3, and P53, in

adductor tissue, at the 2019 and 2020 LTEMP sites (VH, JPO, JB, GoC, GB, and AMT)

compared to the 2012-2015 PWS locations.

4-12 Attachment B

28


adductor tissue, at the 2019 and 2020 LTEMP sites (VH, JPO, JB, GoC, GB, and AMT)

compared to the 2012-2015 PWS locations.

4-12 Attachment B

29

Gill tissue

The 2019 and 2020 transcript profiles of gill tissue from mussels sampled in Port Valdez

and Valdez Arm (LTEMP sites) were statistically compared with transcript profiles of gill tissue

from mussels sampled in LACL and KATM (collected in 2015 and 2016) and PWS (collected in

2012 – 2015). Gene transcript levels associated with contaminant presence that were

significantly different among locations included MT20, Cyp3, and P53 (Figures 15-20). Figure

15, Figure 16, and Figure 17 depict genes in mussel gill tissue primarily associated with

detoxification of contaminants including PAHs, compared among individual LTEMP sites

(AMT, GB, GoC, JB, JPO, and VH). Sites are split into sampling years (if available) 2019 and

2020. The same genes were also compared among broader groupings of sites (LTEMP 2019,

LTEMP 2020, KATM and LACL) (Figure 18, Figure 19, Figure 20).

The following box-and-whisker quartile plots use bars to depict the range from the 10th

to the 90th percentile of individual normalized transcription values for each gene. The

rectangular box indicates the 1st and 3rd quartiles around the median mid-line. Red circles, if

present, represent 5th and 95th percentile outliers. Interpretation of sampling location and gene

abbreviations are provided in Table 1 and Table 2.

4-12 Attachment B

30

Figure 15. MT20 transcript levels in gill tissues collected as part of 2019 and 2020 LTEMP.


gill tissue were found in mussels sampled at GoC19 and VH19. Data were assessed for statistical

significance between locations/years using Kruskal-Wallis with Dunns’ Multiple Comparison

Tests and Bonferroni correction, with reported Z values (NCSS© Statistical Software, 2007,

Kaysville, Utah). “>” can be translated as “has higher transcription than.” Significant differences:

GoC19 > JPO20, VH20; VH19 > VH20 (P = 0.00).

4-12 Attachment B

31

Figure 16. Cyp3 transcript levels in gill tissues collected as part of 2019 and 2020 LTEMP.


levels in gill tissue were found in mussels sampled at AMT 2019, GoC 2019, JPO19, VH19. Data

were assessed for statistical significance between locations/years using Kruskal-Wallis with Dunns’

Multiple Comparison Tests and Bonferroni correction, with reported Z values (NCSS© Statistical


Significant differences: no significant differences.

4-12 Attachment B

32

Figure 17. P53 transcript levels in gill tissues collected as part of 2019 and 2020 LTEMP.

For P53, primarily indicative of cell death and malignant transformation, as well as PAH

exposure, the highest transcript levels in gill tissue were found in mussels sampled in GoC19

and JPO19. Data were assessed for statistical significance between locations/years using

Kruskal-Wallis with Dunns’ Multiple Comparison Tests and Bonferroni correction, with

reported Z values (NCSS© Statistical Software, 2007, Kaysville, Utah). “>” can be translated

as “has higher transcription than.” Significant differences: GoC19 > GB20, JB20, VH20 (P =

0.00).

4-12 Attachment B

33

Figure 18. MT20 transcript levels in gill tissues collected as part of LTEMP 2019 and 2020,

and KATM LACL, and PWS.


gill tissue were found in mussels sampled at the LTEMP 2019 and 2020 sites in comparison

with sites in KATM, LACL, and PWS. Data were assessed for statistical significance between

locations/years using Kruskal-Wallis with Dunns’ Multiple Comparison Tests and Bonferroni

correction, with reported Z values (NCSS© Statistical Software, 2007, Kaysville, Utah). “>” can

be translated as “has higher transcription than.” Significant differences: LTEMP19 > KATM,

LACL, PWS; LTEMP20 > KATM, LACL, PWS; LACL > PWS (P = 0.00).

4-12 Attachment B

34

Figure 19. Cyp3 transcript levels in gill tissues collected as part of LTEMP 2019 and 2020,

and KATM, LACL, and PWS.


levels in gill tissue were found in mussels sampled at the LTEMP 2019 sites in comparison with

LTEMP 2020 and sites in KATM and LACL. Data were assessed for statistical significance

between locations/years using Kruskal-Wallis with Dunns’ Multiple Comparison Tests and

Bonferroni correction, with reported Z values (NCSS© Statistical Software, 2007, Kaysville, Utah).

“>” can be translated as “has higher transcription than.” Significant differences: LTEMP19 >

KATM, LACL, LTEMP20, PWS; LTEMP20 > KATM, PWS; LACL > PWS (P = 0.00).

4-12 Attachment B

35

Figure 20. P53 transcript levels in gill tissues collected as part of LTEMP 2019 and 2020, and

KATM, LACL, and PWS.

For P53, primarily indicative of cell death and malignant transformation, as well as PAH exposure,

the highest transcript levels in gill tissue were found in mussels sampled at the LTEMP 2019 and

2020 sites in comparison with sites in KATM and LACL. Data was assessed for statistical

significance between locations/years using Kruskal-Wallis with Dunns’ Multiple Comparison Tests

and Bonferroni correction, with reported Z values (NCSS© Statistical Software, 2007, Kaysville,

Utah). “>” can be translated as “has higher transcription than.” Significant differences: LTEMP19

> KATM, LACL, PWS; LTEMP20 > KATM, LACL, PWS (P = 0.00).

4-12 Attachment B

36

Three-dimensional scatter plots of the median values for MT20, Cyp3, and P53, in

mussel gill tissue, form a gradient from the 2019 and 2020 LTEMP sites (VH, JPO, JB, GoC,

GB, and AMT), compared to the 2012-2015 PWS, and 2015-2016 KATM and LACL samples

(Figures 21 and 22 differ only in the angle of their orientation). Our results show that the

highest levels of transcription in gill tissues of the three genes are found in mussels sampled at

GoC in 2019 (the yellow cube closest to the axes origin). The lowest levels of transcription for

each of the genes is found in mussels sampled in PWS, LACL, and KATM (blue pyramids),

while transcription of remaining LTEMP samples fell into the higher transcription range.


gill tissue, at the 2019 and 2020 LTEMP sites (VH, JPO, JB, GoC, GB, and AMT) compared

to the PWS, KATM, and LACL locations.

4-12 Attachment B

37


gill tissue, at the 2019 and 2020 LTEMP sites (VH, JPO, JB, GoC, GB, and AMT) compared

to the PWS, KATM, and LACL locations.

Tissue comparison

Three-dimensional scatter plots of the median values for MT20, Cyp3, and P53, in both

mussel gill and adductor tissues from the 2019 and 2020 LTEMP sites, 2012 -2015 PWS, and

2015-2016 KATM and LACL locations show similar but mostly separate response gradients

(Figure 23 and Figure 24). In the scatter plots, gill tissue is represented by cubes and adductor

muscle tissue by spheres. Our results show that the highest levels of transcription of the three

genes are found in gill tissues (cubes), although transcript levels of adductor muscles from

VH19 (red sphere) group with the gill tissues. In the primarily gill cluster, the highest levels of

4-12 Attachment B

38

transcription are found in the LTEMP samples while the lowest levels are found in KATM,

LACL, and PWS. In the adductor cluster (spheres), the highest transcript levels are found in the

LTEMP samples (VH19 is the highest) with lowest samples seen in the PWS samples.


gill and adductor muscle tissue, at the 2019 and 2020 LTEMP sites (VH, JPO, JB, GoC, GB,

and AMT) compared to the PWS, KATM, and LACL locations.

4-12 Attachment B

39


gill and adductor muscle tissue, at the 2019 and 2020 LTEMP sites (VH, JPO, JB, GoC, GB,

and AMT) compared to the PWS, KATM, and LACL locations.

CONCLUSIONS

We have conducted a two-year analysis of gene transcription in two tissues, adductor

muscle and gill, collected from mussels at four sites in Port Valdez in June 2019 and 2020

(AMT, GoC, JPO, VH) with an additional two sites in the Valdez Arm added in June 2020 to

potentially act as clean reference sites (JB, GB). For comparison with these samples, we have

gene expression data for mussels from sites considered to be relatively clean (for adductor

muscle: western PWS Field sites, collected 2012-2015, and for gill: PWS (2012 – 2015), and

KATM and LACL sites, collected 2015-2016). Additionally, we have gene expression data for

4-12 Attachment B

40

adductor tissue in mussels collected from regional harbors (Cordova, Whittier, and Seward) in

2014-2015, representing a group exposed to relatively higher levels of contaminants.

In general, for both adductor muscle and gill, our analyses indicate the expression of

genes most directly associated with contaminant exposure (MT20, Cyp3, P53) was elevated in

the samples from the Valdez area in 2019 and 2020 when compared to samples from “clean”

sites, either in western PWS Field or in KATM and LACL. Additionally, the same gene

expression was also elevated in samples from the Port of Valdez relative to other harbor sites

(Whittier, Seward, and Cordova Harbor). Levels of MT20, Cyp3, and P53 transcripts increase

in response to hydrocarbon exposure. These findings suggest that all six sites sampled in the

Valdez area in 2019 and 2020 have levels of contaminant exposure higher than the

background levels found at more remote sites in PWS, KATM, and LACL. These findings

support the use of gene expression analyses in mussels as a method to monitor the presence of

contaminants in Port Valdez.

RECOMMENDATIONS

1. Add at least one additional site outside Port Valdez to act as a control. When compared

with expression profiles from mussels sampled in KATM, LACL, and PWS, samples from

potential control sites (JB and GB) clustered with samples from the Port Valdez sites

(Figure 13, Figure 14, Figures 21-24). This is an indication that although JB and GB may

be less impacted by hydrocarbon exposure than sites closer to the Valdez Marine

Terminal, they are clearly impacted more than sites in KATM, LACL, and PWS. Thus, for

a broader perspective, it is recommended to add at least one and potentially two control

sites chosen from sites analyzed previously (Bowen et al. 2018). These sites could

potentially be sampled by Gulf Watch Alaska’s environmental monitoring program crews

during yearly monitoring cruises, effectively avoiding any additional field costs.

4-12 Attachment B

41

2. Assess gene transcript levels from all sites at least once per year during the same season;

transcript patterns in mussels have been shown to fluctuate seasonally. However, a more

robust sampling plan and design would help ensure more substantial long-term results.

3. Although overall patterns were similar, transcript profiles were tissue specific to gill

and adductor muscle tissues. To get a more accurate picture of mussel responding to

their environment, dual tissue sampling should continue.

4. Continue to assess hydrocarbon chemistry in mussel tissues. Current levels of

hydrocarbons are below the limits of detection. As per William B. Driskell, consider

adding to the existing panel of hydrocarbon chemistry analyses. It is possible that the

mussels are responding physiologically to a hydrocarbon not being measured.

5. The gene transcript panel was developed prior to pilot trials for PWSRCAC and was

intended to identify a broad range of stimuli. If PWSRCAC decides to include

transcriptomics in future monitoring, we propose modifying this gene panel to

include optimal genes specific to the needs of PWSRCAC. We propose conducting

an experiment with samples from the April 12, 2020, oil spill at the Valdez Marine

Terminal (already collected and in our freezer). We would perform a full

transcriptome analysis (RNAseq) of mussels exposed to the spill. This would allow

for identification of genes specific to the contaminants in a carefully monitored real-

world spill event such as the spill from the Valdez Marine Terminal in April 2020.

This will increase both the specificity and sensitivity of the gene panel for the needs

of the PWSRCAC.

6. Include simple mussel morphometric measurements such as length and height in future

sampling. As PWSRCAC Scientific Advisory Committee member Roger Green stated,

collecting such morphometric measurements is “not advocating for using size, shape,

4-12 Attachment B

42

age or sex as “response to anthropogenic impact” variables; rather one should use

them to control variation in impact which is in fact caused by biological variability in

the animals. All we are talking about is good study design which minimizes “noise”

due to factors other than the contaminants. It is similar to maintaining consistency of

field and lab procedure across locations.”

4-12 Attachment B

43

REFERENCES

Balseiro P, Falcó A, Romero A, Dios S, Martínez-López A, Figueras A, Estepa A, Novoa B. 2011.

Mytilus galloprovincialis Myticin C: a chemotactic molecular with antiviral activity and

immunoregulatory properties. PLOS ONE 6:e23140 DOI 10.1371/journal.pone.0023140.

Banni M, Dondero F, Jebali J, Guerbej H, Boussetta H, Viarengo A. 2007. Assessment of heavy

metal contamination using real-time PCR analysis of mussel metallothionein mt10 and

mt20 expression: a validation along the Tunisian coast. Biomarkers 12:369_383 DOI

10.1080/13547500701217061.

Banni M, Negri A, Rebelo M, Rapallo F, Boussetta H, Viarengo A, Dondero F. 2009. Expression

analysis of the molluscan p53 protein family mRNA in mussels (Mytilus spp.) exposed to

organic contaminants. Comparative Biochemistry and Physiology Part C: Toxicology &

Pharmacology 149:414_418 DOI 10.1016/j.cbpc.2008.09.017.

Banni M, Negri A, Mignone F, Boussetta H, Viarengo A, Dondero F. 2011. Gene expression

rhythms in the mussel Mytilus galloprovincialis (Lam.) across an annual cycle. PLOS ONE

6:e18904 DOI 10.1371/journal.pone.0018904.

Banni M, Sforzini S, Arlt VM, Barranger A, Dallas LJ, Oliveri C, Aminot Y, Pacchioni B,

Millino C, Lanfranchi G, Readman JW, Moore MN, Viarengo A, Jha AN. 2017.

Assessing the impact of Benzo [a] pyrene on marine Mussels: Application of a novel

targeted low density microarray complementing classical biomarker responses. PloS

ONE 12: e0178460.

Bowen L, Miles AK, Ballachey B, Waters S, Bodkin J, Lindeberg M, Esler D. 2018. Gene

transcription patterns in response to low level petroleum contaminants in Mytilus

trossulus from field sites and harbors in southcentral Alaska. Deep Sea Research Part II,

147: 27-35.

4-12 Attachment B

44

Chen ZF, Wang H, Matsumura K, Qian PY. 2012. Expression of calmodulin and myosin light

chain kinase during larval settlement of the barnacle Balanus amphitrite. PLOS ONE

7:e31337 DOI 10.1371/journal.pone.0031337.

Counihan K, Bowen L, Ballachey B, Coletti H, Hollmen T, Pister B. 2019. Physiological and

gene transcription assays to assess responses of mussels to environmental changes. PeerJ

7: e7800.

De Maio AM. 1999. Heat shock proteins: facts, thoughts, and dreams. Shock 11:1_2.

Farr S, Dunn RT. 1999. Concise review: gene expression applied to toxicology. Toxicological

Sciences 50: 1-9.

Fukuda R, Zhang H, Kim JW, Shimoda L, Dang CV, Semenza GL. 2007. HIF-1 regulates

cytochrome oxidase subunits to optimize efficiency of respiration in hypoxic cells.

Cell 129:111_122 DOI 10.1016/j.cell.2007.01.047.

Giuliani ME, Benedetti M, Arukwe A, Regoli F. 2013. Transcriptional and catalytic responses of

antioxidant and biotransformation pathways in mussels, Mytilus galloprovincialis, exposed

to chemical mixtures. Aquatic Toxicology 134_135:120_127 DOI

10.1016/j.aquatox.2013.03.012.

Goodson MS, Crookes-Goodson WJ, Kimbell JR, McFall-Ngai MJ. 2006. Characterization and

role of p53 family members in the symbiont-induced morphogenesis of the Euprymna

scolopes light organ. The Biological Bulletin 211:7_17 DOI 10.2307/4134573.

Heid, CA, Stevens J, Livak KJ, Williams PM. 1996. Real time quantitative PCR. Genome

Research 6: 986-994.

Hüning AK, Melzner F, Thomsen J, Gutowska MA, Kramer L, Frickenhaus S, Rosenstiel P,

Pörtner HO, Philipp EER, Lucassen M. 2013. Impacts of seawater acidification on mantle

4-12 Attachment B

45

gene expression patterns of the Baltic Sea blue mussel: implications for shell formation and

energy metabolism. Marine Biology 160:1845_1861 DOI 10.1007/s00227-012-1930-9.

Iwama GK, Mathilakath MV, Forsyth RB, Ackerman PA. 1999. Heat shock proteins and

physiological stress in fish. American Zoology 39:901_909

DOI10.1093/icb/39.6.901.

Kadota K, Nishimura SI, Bono H, Nakamura S, Hayashizaki Y, Okazaki Y, Takahashi K. 2003.

Detection of genes with tissue-specific expression patterns using Akaike’s information

criterion procedure. Physiological Genomics 12: 251-259.

Li S, Xie L, Zhang C, Zhang Y, Gu M, Zhang R. 2004. Cloning and expression of a pivotal

calcium metabolism regulator: calmodulin involved in shell formation from pearl oyster

(Pinctada fucata). Comparative Biochemistry and Physiology Part A: Molecular &

Integrative Physiology 138:235_243 DOI 10.1016/j.cbpc.2004.03.012.

Li S, Huang J, Liu C, Liu Y, Zheng G, Xie L, Zhang R. 2016. Interactive effects of seawater

acidification and elevated temperature on the transcriptome and biomineralization in the

pearl oyster Pinctada fucata. Environmental Science & Technology 50:1157_1165 DOI

10.1021/acs.est.5b05107.

McLoughlin K, Turteltaub K, Bankaitis-Davis D, Gerren R, Siconolfi L, Storm K, Cheronis J,

Trollinger D, Macejak D, Tryon V, Bevilacqua M. 2006. Limited dynamic range of

immune response gene expression observed in healthy blood donors using RT-PCR.

Molecular Medicine 12: 185-195.

Mitta G, Vandenbulcke F, Hubert F, Salzet M, Roch P. 2000. Involvement of mytilins in mussel

antimicrobial defense. Journal of Biological Chemistry 275:12954_1262 DOI

10.1074/jbc.275.17.12954.

4-12 Attachment B

46

Parisi MG, Toubiana M, Mangano V, Parrinello N, Cammarata M, Roch P. 2012. MIF from

mussel: coding sequence, phylogeny, polymorphism, 3D model and regulation of

expression. Developmental & Comparative Immunology 36:688_696 DOI

10.1016/j.dci.2011.10.014.

Philipp EER, Kraemer L, Melzner F, Poustka AJ, Thieme S, Findeisen U, Schreiber S, Rosenstiel

P. 2012. Massively parallel RNA sequencing identifies a complex immune gene repertoire

in the lophotrochozoan Mytilus edulis. PLOS ONE 7:e33091 DOI

10.1371/journal.pone.0033091.

Poynton HC, Vulpe CD. 2009. Ecotoxicogenomics: emerging technologies for emerging

contaminants. Journal of the American Water Resources Association 45: 83-96.

Romero A, Estévez-Calvar N, Dios S, Figueras A, Novoa B. 2011. New insights into the apoptotic

process in mollusks: characterization of caspase genes in Mytilus galloprovincialis. PLOS

ONE 6:e17003 DOI 10.1371/journal.pone.0017003.

Tsan MF, Gao B. 2004. Cytokine function of heat shock proteins. American Journal of Physiology-

Cell Physiology 286:C739_C744 DOI 10.1152/ajpcell.00364.2003.

Wu R. 2002. Hypoxia: from molecular responses to ecosystem responses. Marine Pollution

Bulletin 45:35_45 DOI 10.1016/S0025-326X(02)00061-9.

4-12 Attachment B

47

APPENDIX

Table 3. Medians and ranges calculated for mussels (adductor muscle tissue) sampled at LTEMP sites in 2019 and 2020, as well

as from PWS sites from 2012-2015.

AMT19 AMT20 GB20 CoC19 GoC20 JB20

Median Range Median Range Median Range Median Range Median Range Median Range

CaM 19.58 4.41 19.05 3.96 19.58 4.41 19.05 3.96 20.82 3.57 19.87 2.91

Casp8 14.11 3.41 14.54 4.29 14.11 3.41 14.54 4.29 15.63 4.77 13.33 2.28

MIF 19.59 8.99 20.23 3.91 19.59 8.99 20.23 3.91 18.56 11.01 20.60 7.99

Calponin 19.94 3.62 20.07 4.86 19.94 3.62 20.07 4.86 19.28 4.14 20.44 4.55

Chitinase 19.15 3.02 19.06 3.55 19.15 3.02 19.06 3.55 20.58 4.12 20.10 2.79

CCOIV 21.32 7.37 20.27 3.18 21.32 7.37 20.27 3.18 22.59 5.28 21.68 6.91

HSP70 14.06 5.98 11.55 2.99 14.06 5.98 11.55 2.99 13.75 4.66 13.44 4.21

HSP90 13.73 5.12 14.01 5.55 13.73 5.12 14.01 5.55 15.02 6.17 14.17 5.70

HIFa 15.65 2.30 14.64 3.78 15.65 2.30 14.64 3.78 16.14 4.27 16.37 3.46

MyticinB 16.34 6.49 14.88 12.20 16.34 6.49 14.88 12.20 16.45 12.29 13.93 11.16

Mytilin 18.73 3.70 17.52 6.91 18.73 3.70 17.52 6.91 17.96 8.15 16.86 7.03

MT20 9.85 2.83 8.87 2.58 9.85 2.83 8.87 2.58 10.19 5.25 9.45 3.93

Cyp3 17.30 3.49 16.13 3.69 17.30 3.49 16.13 3.69 17.11 4.76 15.98 4.32

P53 16.34 3.10 15.77 4.15 16.34 3.10 15.77 4.15 17.53 3.29 16.21 2.54 JPO19 JPO20 VH19 VH20 PWS

Median Range Median Range Median Range Median Range Median Range

CaM 21.24 11.09 20.32 6.43 16.82 5.04 19.92 5.56 19.96 9.60

Casp8 15.34 9.95 14.12 3.47 12.14 5.27 13.92 5.96 16.66 10.08

MIF 17.77 17.62 20.01 7.40 14.52 10.31 17.57 7.51 18.71 13.60

Calponin 21.35 10.38 18.36 5.05 18.40 5.33 18.89 5.57 21.58 32.39

Chitinase 21.24 7.94 19.65 3.65 18.11 1.80 20.48 5.32 20.27 23.04

CCOIV 21.99 9.97 21.20 6.94 17.83 11.78 21.06 8.14 22.11 14.28

HSP70 12.59 9.04 13.08 3.94 9.79 3.57 12.99 6.23 12.38 16.46

HSP90 13.12 9.49 14.96 4.18 13.28 4.58 14.29 6.16 14.36 11.64

HIFa 14.07 8.18 16.37 3.28 13.48 1.48 15.87 4.51 15.67 7.50

MyticinB 17.75 12.29 16.20 10.92 9.15 8.56 15.10 9.65 14.76 17.04

Mytilin 19.78 10.96 17.94 12.93 14.67 8.14 17.87 5.39 18.71 13.30

MT20 9.95 12.36 9.90 1.97 6.84 6.03 10.59 3.96 14.84 11.07

Cyp3 17.27 13.13 15.71 3.03 13.42 4.76 16.40 5.51 17.74 9.64

P53 15.83 10.17 16.58 3.15 13.15 4.28 16.07 5.30 18.61 28.03

4-12 Attachment B

48

Table 4. Medians and ranges calculated for mussels (gill tissues) sampled at LTEMP sites in 2019 and 2020, as well as from other

studies in KATM and LACL (2015-2016) and PWS (2012-2015).

AMT19 AMT20 GB20 GoC19 GoC20 JB20 JPO19

Median Range Median Range Median Range Median Range Median Range Median Range Median Range

CaM 17.11 5.11 16.76 4.76 17.08 2.63 14.61 1.91 16.57 2.74 16.63 2.35 17.05 3.10

Casp8 9.47 3.54 8.80 2.65 9.74 3.28 8.62 2.55 9.73 4.94 9.46 1.82 9.59 3.34

MIF 17.36 9.52 17.00 8.00 18.75 8.49 17.75 2.88 16.48 9.81 18.75 7.89 16.33 14.68

Calponin 25.87 3.65 23.54 2.36 24.23 2.50 23.15 4.43 23.81 3.76 24.64 2.86 26.22 4.75

Chitinase 20.90 5.08 20.04 5.89 19.82 4.54 19.57 5.41 21.15 5.37 20.15 3.97 21.42 3.39

CCOIV 21.00 6.92 17.04 8.98 18.79 5.21 16.71 5.50 19.48 5.26 19.05 7.63 18.51 4.48

HSP70 11.87 6.04 12.21 5.94 13.03 9.98 10.25 1.78 11.90 4.03 13.02 4.53 11.93 6.07

HSP90 13.39 4.34 12.72 5.19 12.51 4.19 13.80 5.88 13.34 4.17 12.82 5.33 12.10 6.86

HIFa 13.52 2.50 14.24 3.12 13.20 3.97 13.01 2.76 13.90 4.24 13.74 4.61 12.73 3.78

MyticinB 9.01 13.36 13.10 10.26 12.79 8.03 10.93 9.51 12.64 13.73 12.75 11.24 15.05 12.36

Mytilin 15.79 4.26 14.26 2.99 14.88 3.79 14.35 5.01 14.03 8.98 15.13 6.67 16.10 4.44

MT20 6.51 4.35 7.11 5.02 6.57 3.11 5.24 1.62 6.91 5.72 6.80 2.08 5.99 7.19

Cyp3 12.53 3.67 13.58 3.98 14.15 3.57 12.98 4.22 15.33 5.11 13.60 3.72 13.31 4.17

P53 11.00 2.23 10.97 1.98 11.96 2.03 10.37 2.09 11.31 2.54 11.80 2.14 10.81 6.20 JPO20 VH19 VH20 PWS KATM LACL

Median Range Median Range Median Range Median Range Median Range Median Range

CaM 17.26 7.33 14.79 3.09 16.77 6.29 17.06 6.02 16.85 7.89 15.77 8.72

Casp8 9.63 4.08 9.51 4.30 9.46 4.65 11.79 5.89 10.60 5.56 10.27 9.89

MIF 18.09 6.48 14.22 8.17 15.87 12.12 19.49 8.93 18.06 10.31 17.84 11.30

Calponin 24.35 3.66 23.44 3.43 24.20 4.29 25.59 6.49 25.21 6.76 24.56 10.29

Chitinase 20.67 3.64 19.51 7.96 20.44 4.88 19.17 3.62 18.55 11.94 18.69 7.97

CCOIV 18.53 5.76 17.01 8.00 18.47 6.27 18.94 11.89 18.31 11.59 18.42 11.22

HSP70 12.51 5.32 10.47 4.21 12.21 4.79 11.92 5.70 12.13 8.59 11.03 11.37

HSP90 14.52 4.08 13.55 6.83 13.89 5.96 14.19 5.18 13.97 8.58 13.20 6.22

HIFa 13.80 3.90 12.84 6.17 14.28 3.11 14.83 3.42 14.33 3.87 13.89 5.78

MyticinB 12.89 10.03 7.14 8.64 12.50 13.58 11.79 13.42 13.73 13.74 11.56 15.05

Mytilin 15.04 10.83 13.86 6.37 14.66 6.22 15.94 8.62 16.40 10.25 15.54 8.76

MT20 6.93 4.19 5.44 3.02 7.61 6.35 12.77 10.20 10.60 13.46 8.83 14.51

Cyp3 13.66 4.39 13.39 2.97 14.16 5.94 15.79 7.10 14.96 5.61 13.95 11.44

P53 11.45 3.03 11.44 2.09 11.96 3.91 13.83 5.53 13.01 9.51 12.68 12.53

4-12 Attachment B

report acceptance: 2020 ltemp sampling & mussel

Documents