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Page 1: Earthworms as Ecoengineers Following Remediation

Earthworms as Ecoengineers in the Restoration of Oil and Brine-

Impacted Soils Following RemediationNitya AlahariKerry Sublette

Eleanor JenningsCenter for Applied Biogeosciences

University of TulsaGreg ThomaDuane Wolf

University of ArkansasKathleen Duncan

University of OklahomaTim Todd

Kansas State UniversityMac A. Callaham, Jr.USDA-Forest Service

Page 2: Earthworms as Ecoengineers Following Remediation

Remediation of Oil and Brine Spills

• Oil– Fertilizer

• Increase rates• Prevents depletion of soil nutrient pool

– Organic matter• Increases O2 infiltration• Decay products help rebuild soil structure• Substrate for soil fauna

– Tilling• Aeration• Mixing• Distribute oil in the soil to create more oil-water interface

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Remediation of Oil and Brine Spills

• Brine– Organic matter

• Increases permeability to water• Decay products help rebuild soil structure• Substrate for soil fauna

– Tilling• Mixing• Improving permeability

– Fertilizer• Promote biodegradation of organic matter• Prevents impact on soil nutrient pool

– Gypsum• Combat sodicity

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Restoration of Oil- and Brine-impacted Sites

• Both the original spill and the remediation process disrupt soilecology– Disruptions in N and P cycling– Reduced diversity of soil microbes and invertebrates– Loss of vegetation

• All levels of ecosystem affected– Producers– Consumers– Decomposers

• Is restoration of the soil ecosystem the real definition of “clean” for a high value site?– Left to nature restoration is a lengthy process

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Increasing the Rate of Restoration of Soil Ecosystems

• Are earthworms the answer?– Earthworm castings

• contain higher concentrations of SOM and bioavailable nutrients than the surrounding bulk soil

• exhibit greater microbial activity and higher rates of respiration than bulk soil

• lead to the formation of stable soil aggregates which increase the permeability of the soil to air and water

– Earthworm burrows create pathways for root growth, water movement, and nutrient transport

– Earthworm-related effects stimulate the uptake of nutrients by plants which results in increased growth rates of plants and greater levels of biomass

– All of these effects are in proportion to the density of earthworms in the soil and can persist for long periods of time

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Project Objectives

• Determine the appropriate amendments to optimize the re-introduction of earthworms to oil- and brine-impacted sites which have been remediated but not fully restored.

• These data will – Lead to a cost-effective protocol for re-

introduction and cultivation of earthworms in these sites

– Demonstrate the benefits of earthworm re-introduction on re-vegetation of these sites in terms of increased plant biomass and greater species diversity.

Page 7: Earthworms as Ecoengineers Following Remediation

Previous Work (Callaham et al., 2002*)

• Greenhouse study of the survival and effects of earthworms (Eisenia fetida) in landfarm soil containing TPH concentrations averaging 33,000 mg/kg.

• Results:– earthworms will survive in bioremediated soil with high

residual TPH concentrations; – organic matter is necessary for their long-term survival; – earthworm activity resulted in greater accumulation of

above- and below-ground plant biomass.

*Env. Toxicology and Chem., 21, 1658-1663 (2002)

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Added NaCl(g/kg of

soil)

Fraction of replicate microcosms showing

evidence of reproduction*

Mean survival(%)

0 4/4 90.0

1 4/4 95.0

3 2/4 90.0

5 0/4 97.5

7.5 0/4 95.0

10 0/4 95.0

15 0/4 90.0

*Art Stewart (Oak Ridge National Lab)

Results of 17-d Test to Determine Sensitivity of the Earthworm Eisenia

fetida to NaCl in soil*

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Test Sites

• G7– 2000 spill of produced fluids (W/O ratio of 10-15)– Four treatments: combinations of hay, fertilizer (13:13:13),

and no treatment– Treatment terminated in 2004

• LF– Site of crude oil landfarm closed in 1997– Final TPH (EPA 418.1) < 9000 mg/kg

Page 10: Earthworms as Ecoengineers Following Remediation

Treatments / Experimental Design

• Worms only• Fertilizer only• Hay only• Worms + Hay• Worms + Fertilizer• Fertilizer + Hay• Worms + Hay

+Fertilizer• No treatment

• Four blocks each site

• Four replicates of each treatment in each block

• Sacrificial sampling of one replicate of each treatment per block per site

Page 11: Earthworms as Ecoengineers Following Remediation

BlockG7

Na+

(mg/kg)N=3

Cl-(mg/kg)N=3

BlockLF

TPH*(mg/kg)N=4

1 711 ± 198 900 ± 298 5 11546 ± 24042 652 ± 39 788 ± 94 6 16634 ± 21843 633 ± 201 576 ± 171 7 9535 ± 19034 567 ± 79 301 ± 84 8 16511 ± 5350

Initial Test Site Conditions

*CH2Cl2 extractables (gravimetric)

TPH (EPA 418.1) < 9000 mg/kg

Page 12: Earthworms as Ecoengineers Following Remediation

Block 1 Block 2 Block 3 Block 4

G7

Worms + Hay + Fertilizer

Worms + Hay

Worms + Fertilizer

Hay + Fertilizer

Worms

Hay

Fertilizer

No Treatment

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G7

Page 14: Earthworms as Ecoengineers Following Remediation

LF

Page 15: Earthworms as Ecoengineers Following Remediation

Project Timeline

• Rip and till sites; homogenize to extent possible• Install earthworm enclosures and add amendments

(fertilizer and/or hay)– G7: May 2, 2005 – LF: May 31, 2005

• Inoculate with Eisenia fetida ( 5 worms per enclosure per worm treatment); cover with panty hose– G7: June 23, 2005– Lf: July 7, 2005

• First sampling– G7: July 21, 2005– LF: August 2, 2005

• Second sampling– G7: Oct. 15, 2005– LF: Oct. 14, 2005

Page 16: Earthworms as Ecoengineers Following Remediation

Why Eisenia fetida?

• Readily available commercially all over the U.S. for a reasonable cost ($15-$20/1000 worms).

• Easily cultivated by inexperienced personnel

• Requires high concentrations of soil organic matter and is likely to be replaced by indigenous species when they begin to migrate into the restored sites

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Both sites ripped to 12” and tilledG7

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LF

Page 19: Earthworms as Ecoengineers Following Remediation

Homogenizing soil from Block 5 for earthworm enclosures at LF

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Filling enclosure with homogenized soil at LF

Page 21: Earthworms as Ecoengineers Following Remediation

Earthworm enclosures installed and amendments added at LF

Page 22: Earthworms as Ecoengineers Following Remediation

Entire site covered with hay for moisture and temperature control

Page 23: Earthworms as Ecoengineers Following Remediation

Sampling enclosure; note that space surrounding enclosure has also been sampled

Page 24: Earthworms as Ecoengineers Following Remediation

Site Maintenance

• Barb wire fence to keep out buffalo; electric fence to keep out coyotes

• Each site watered every other day unless there was sufficient rain

Page 25: Earthworms as Ecoengineers Following Remediation

Spring Fall Spring FallNutrients X X X XBrine X X X XTPH X X X XPLFA X X X XDNA XN cycling bacteria

X X X X

Nematodes X X X XPlants X X XEarthworms X X X X

Sampling and Analysis

Page 26: Earthworms as Ecoengineers Following Remediation

G7 Worm Count (July 21, 2005)

F FH H N W WF WFH WH

Treatment

0

2

4

6

8

10

12

14W

orm

Cou

nt

E. fetida A. trapezoides Diplocardia sp.

E. fetida is missing in action (mostly)

Local species have moved in – more so in G7 than LF

Page 27: Earthworms as Ecoengineers Following Remediation

0

2

4

6

8

10

12

16-18 18-20 20-22 22-24 24-26 26-28 28-30 30-32

Percent Moisture

No.

of O

bser

vatio

ns o

f W

orm

s

G7LF

Frequency of worm observations related to soil moisture

On average soil moistures in LF were 4% lower than in G7 (avg. 26%)

Page 28: Earthworms as Ecoengineers Following Remediation

FH F H N

G7 Modified Treatment

20

22

24

26

28

30%

Moi

stur

e Mean Mean±SE

*If native worms are widely distributed at the sites and enter and exit enclosures freely, then there are only four treatments

Page 29: Earthworms as Ecoengineers Following Remediation

FH F H N

G7 Modified Treatment

5

10

15

20

25

30

35

40

45P

(mg/

kg)

Mean Mean±SE a a

b b

Page 30: Earthworms as Ecoengineers Following Remediation

FH F H N

G7 Modified Treatment

4

6

8

10

12

14

16

18NH

4-N

(mg/

kg)

Mean Mean±SE a a

bb

Simple decomposition of hay with a C/N of 25-30 should result in net immobilization

Page 31: Earthworms as Ecoengineers Following Remediation

FH F H N

G7 Modified Treatment

1.0

1.2

1.4

1.6

1.8

2.0

2.2

2.4NO

3-N

(mg/

kg)

Mean Mean±SE

aa

ab

b

Page 32: Earthworms as Ecoengineers Following Remediation

FH F H N

G7 Modified Treatment

1.30

1.35

1.40

1.45

1.50

1.55

1.60

1.65

1.70

1.75

1.80To

tal C

arbo

n (%

) Mean Mean±SE

aa

a

a

p=0.31

Page 33: Earthworms as Ecoengineers Following Remediation

FH F H N

G7 Modified Treatment

0.096

0.100

0.104

0.108

0.112

0.116

0.120

0.124To

tal N

(%)

Mean Mean±SE

aa

aa

p=0.27

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FH F H N

LF Modified Treatment

20

30

40

50

60

70

80

90

100

110

120P

(mg/

kg)

Mean Mean±SE

a

a

b

b

Page 35: Earthworms as Ecoengineers Following Remediation

FH F H N

G7 Modified Treatment

0

100

200

300

400

500

600

700

800To

tal N

emat

odes

(# /

100

g)

Mean Mean±SE

a

b

b b

Page 36: Earthworms as Ecoengineers Following Remediation

FH F H N

G7 Modified Treatment

1.3

1.4

1.5

1.6

1.7

1.8

1.9

2.0

2.1N

emat

ode

Mat

urity

Inde

x Mean Mean±SE

a

ab

b

b

Page 37: Earthworms as Ecoengineers Following Remediation

Preliminary Conclusions

• Earthworms will invade and survive in remediated oil- or brine-impacted soil– organic matter– moisture

• Earthworm activity increases bioavailability of nutrients in these damaged sites (?)

Page 38: Earthworms as Ecoengineers Following Remediation

1 2 3 4

Block

0

5

10

15

20

25

30G

7 W

orm

Bur

row

s(1

ft2 d

iam

eter

circ

le)

Oct

ober

200

5 Mean Mean±SE

Page 39: Earthworms as Ecoengineers Following Remediation

5 6 7 8Block

0

5

10

15

20

25

30LF

Wor

m B

urro

ws

(1 ft

2 dia

met

er c

ircle

)O

ctob

er 2

005

Mean Mean±SE

Page 40: Earthworms as Ecoengineers Following Remediation

Acknowledgement

This work was funded by

the Integrated Petroleum Environmental Consortium (IPEC)

Page 41: Earthworms as Ecoengineers Following Remediation

Extra Slides

Page 42: Earthworms as Ecoengineers Following Remediation

WFH WF WH W FH F H N

Treatment

0

10

20

30

40

50

60P

(mg/

kg)

Mean Mean±SE

Page 43: Earthworms as Ecoengineers Following Remediation

WFH WF WH W FH F H N

Treatment

2

4

6

8

10

12

14

16

18

20

22

24N

H4-

N (m

g/kg

) Mean Mean±SE

Page 44: Earthworms as Ecoengineers Following Remediation

WFH WF WH W FH F H N

Treatment

21

22

23

24

25

26

27

28

29

30%

Moi

stur

e Mean Mean±SE

Page 45: Earthworms as Ecoengineers Following Remediation

WFH WF WH W FH F H N

Treatment

0

200

400

600

800

1000

1200

1400To

tal N

emat

odes

(# /

100

g)

Mean Mean±SE

Page 46: Earthworms as Ecoengineers Following Remediation

WFH WF WH W FH F H N

Treatment

1.2

1.4

1.6

1.8

2.0

2.2

2.4N

emat

ode

Mat

urity

Inde

x

Mean Mean±SE

Page 47: Earthworms as Ecoengineers Following Remediation

WFH WF WH W FH F H N

Treatment

200

250

300

350

400

450

500

550EC

(1:1

, um

hos/

cm)

Mean Mean±SE

Page 48: Earthworms as Ecoengineers Following Remediation

WFH WF WH W FH F H N

G7 Treatment

1.2

1.3

1.4

1.5

1.6

1.7

1.8

1.9To

tal C

arbo

n (%

)

Mean Mean±SE

Page 49: Earthworms as Ecoengineers Following Remediation

G7 July 21, 2005 Sampling

WFH WF WH W FH F H N

Treatment

10

12

14

16

18

20

22

24

26

28

30%

Moi

stur

e

ab ab ab b ab ac c ac

Page 50: Earthworms as Ecoengineers Following Remediation

LF August 2, 2005 Sampling

WFH WF WH W FH F H N

Treatment

10

12

14

16

18

20

22

24

26

28

30%

Moi

stur

e

a a a a a a a b


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