instituto de recursos naturales y agrobiología de sevilla
DESCRIPTION
José Julio Ortega. Instituto de Recursos Naturales y Agrobiología de Sevilla. Consejo Superior de Investigaciones Científicas. ASSESSING THE MICROBIAL AVAILABILITY OF POLYCYCLIC AROMATIC HYDROCARBONS PRESENT IN MARINE SPILLS. OBJECTIVES. - PowerPoint PPT PresentationTRANSCRIPT
Instituto de Recursos Naturales y Agrobiología de SevillaConsejo Superior de Investigaciones Científicas
José Julio Ortega
ASSESSING THE MICROBIAL AVAILABILITY OF POLYCYCLIC AROMATIC HYDROCARBONS PRESENT IN
MARINE SPILLS
Proyect VEM 2004-08556: “Microbial availability and metabolism of polycyclic aromatic hydrocarbons present in marine oil spills. Implications for their natural attenuation and bioremediation”
Examine PAH bioavailability and metabolism in different scenarios relevant to marine oil spills: water column (dispersants), shoreline (bioremediation) and sediments (nat. attenuation)
OBJ. 1 Microbial physiology and metabolism of PAH biodegradation in marine microorganisms
OBJ. 2 Study of physicochemical processes involved in PAH bioavailability and theirmodification by biological factors and spill management strategies
OBJ. 3 Bioavailability and metabolism of PAH in microcosms
OBJECTIVES
Dissolved in macropores
Sorbed to organic matter and clay
fractions1
Sorbed in micropores smaller than bacteria
2
Dissolved in non-aqueous phase
liquids
3
LOW BIOAVAILABILITY
(slow kinetics, residual fractions)
BIOSURFACTANT PRODUCTION
RE (g/ml)0 50 100 150
Sur
face
tens
ion
(mN
/m)
30
40
50
60
70
80
CMC
Effect of Pseudomonas aeruginosa 19SJ biosurfactants on surface tension: critical micelle concentration
Minutes0 20 40 60
Phe
nan
thre
ne
(g/
ml)
0
1
2
3
4
60
01
25
156
Growth and biosurfactant production byPseudomonas aeruginosa 19SJ from solid phenanthrene (10 mg/ml)
Hours
0 200 400 600 800
log
CF
U
7,00
7,75
8,50
9,25
RE
(µ
g/m
l)
0
50
100
150
200
250
M. Garcia-Junco et al., Environ. Microbiol. 2001, 3, 561-569
Effect of biosurfactants from Pseudomonas aeruginosa 19SJ on partitioning of pyrene from a NAPL
Biosurf. Solids Part. rate CeqCeqsolids
(µg/mL) (mg/mL) (ng/mL/h) (ng/mL) (ng/mg)
0
10
100
0
100
0
0
0
1
1
0,5
1,6
11,6
0,7
27,4
18
85
547
27
650
-
-
-
16
40
M. Garcia-Junco et al., Environ. Sci. Technol. 2003, 37, 2988-2996
CHEMOTAXIS
CHEMOTAXISCAPILLARY METHOD
slide
coverslip
Bacterial suspension
Chemoattractant or control
Capillary tube
1 L
Chemotaxis towards phenanthrene of Pseudomonas putida 10D
J.J. Ortega-Calvo et al., FEMS Microbiol. Ecol. 2003, 44, 373-381
0,0
0,2
0,4
0,6
0,8
1,0
1,2
1,4
1,6
control 0,32 0,65 1,29
Phen conc (µg/ml)
Sp
eed
(m
m/m
in)
Environmental sample
TENAX
SAMPLESUSPENSION
DESORPTION BIODEGRADATION
SAMPLESUSPENSION
NaOH TRAP
- TENAX EXTRACTION- HPLC ANALYSIS
- MC EXTRACTION- HPLC ANALYSIS (native PAH)- 14CO2 MEASUREMENTS (14C-PAH)
BIOAVAILABILITY ASSAY
250 ml erlenmeyer flask
B. HPLC analysis
40 g Sample
Teflon-lined stopper
Lateral body
A. 14CO2 production
Syringe sampling
Alkali trap40 g sample + radiolabelled PAH
Main body
Teflon-lined stoppers
Biometric flask
14CO2
St / S0 = Frap * exp (-Krap * t) + Fslow * exp (-Kslow * t)
Tiempo (h)
0 200 400 600 800 1000
Ln St /
S 0
-5
-4
-3
-2
-1
0
Tiempo (h)
0 200 400 600 800 1000
Ln St /
S0
-6
-5
-4
-3
-2
-1
0
Flu Fen
Tiempo (h)
0 200 400 600 800 1000
Ln St /
S0
-4
-3
-2
-1
0
Tiempo (h)
0 200 400 600 800 1000
Ln S
t / S
0
-6
-5
-4
-3
-2
-1
0
AntFlut
Tiempo (h)
0 200 400 600 800 1000
Ln S
t / S
0
-5
-4
-3
-2
-1
0
Tiempo (h)
0 200 400 600 800 1000
Ln S
t / S
0
-4
-3
-2
-1
0
Pyr Benz(a)pyr
DESORPTION OF PAHs WITH TENAX
CREOSOTE-POLLUTED SOIL
DESORPTION & BIODEGRADATION
Time (h)
0 200 400 600 800 1000
Ln St /
S0
-6
-5
-4
-3
-2
-1
0
Phen desorpPhen biodeg
Time (h)
0 200 400 600 800 1000
Ln S
t / S
0
-4
-3
-2
-1
0
BaP desorpBaP biodeg
Phenanthrene Benz(a)pyrene Tenax Biodegradation Tenax Biodegradation
Frap (%) 96,1 94,9 86,7 31,4
Fslow (%) 3,9 5,1 13,3 68,6
Krap (h-1) 0,38 0,15 0,11 0,03
Kslow (x10-3 h-1) 2,70 2,20 1,80 0,08
St / S0 = Frap * exp (-Krap * t) + Fslow * exp (-Kslow * t)
Tiempo (h)
0 200 400 600 800 1000
Ln St /
S 0
-0,5
-0,4
-0,3
-0,2
-0,1
0,0
Ant
Tiempo (h)
0 200 400 600 800 1000
Ln St /
S 0
-0,5
-0,4
-0,3
-0,2
-0,1
0,0
Fen
Tiempo (h)
0 200 400 600 800 1000
Ln St /
S 0
-0,5
-0,4
-0,3
-0,2
-0,1
0,0Fluor
Tiempo (h)0 200 400 600 800 1000
Ln St /
S 0
-0,5
-0,4
-0,3
-0,2
-0,1
0,0
Pyr
Tiempo (h)
0 200 400 600 800 1000
Ln St /
S 0
-0,5
-0,4
-0,3
-0,2
-0,1
0,0Benz(a)ant
Tiempo (h)
0 200 400 600 800 1000
Ln St /
S 0
-0,5
-0,4
-0,3
-0,2
-0,1
0,0Benz(a)pyr
DESORPTION OF PAHs WITH TENAX
BIOREMEDIATED SOIL
Time (days)
0 5 10 15 20 25 30 35 40 45
% 1
4 C m
ine
rali
zed
0
10
20
30
40
50
60
70
80
HA
P (
mg
/Kg
)
0
20
40
60
80
Fen Ftno Pir
31.49 %
23.80 %0.35 %
BIOAVAILABILITY OF PAH IN BIOREMEDIATED SOIL (SOILREM E6068)(0.2 g/Kg PAH)
Fen
Pir
Ftno
42
98 186
1366
1250
372
118
404
59111
85
346142
222
16612455
429
GIBRALTAR
SAMPLING POINTS
ROADS
URBAN ZONES
INDUSTRIAL ZONES
Background PAH pollution in Gibraltar Area (Cádiz)(µg/kg 16 EPA PAH)
Phenanthrene content in soils from Gibraltar Area
*Wild, S.R. & Jones, K.C., Environ. Pollut. 1995, 88, 91-108
% Organic matter
0 1 2 3 4 5 6 7 8 9 10 11
µg
/kg
ph
en
an
thre
ne
0
20
40
60
80
100
120
Average: 40.6 µg/kg
UK rural soils: 14 µg/kg*
Bioaccessibility of native phenanthrene to autochtonous microbial populations in soils with different % OM
Time (days)
0 20 40 60 80 100 120
% 14
C m
iner
aliz
ed
0
10
20
30
40
50
1.72
10.04 4.504.55
2.01
Conditions: • 40 g soil amended with 11.8 µg/kg 14C-phen dissolved in distilled water
14C
% OM
Ph
e (
g/k
g)
0
20
40
60
80
100
120
140
160
total concentrationnot bioaccessible
1.72 2.01 4.5 4.55 10.04
31.6 %17.4 %
24.6 %
native
CONCLUSIONS
1. FAR FROM BEHAVING AS PASSIVE CATALISTS, MICROORGANISMS INHABITING PAH-POLLUTED SOILS AND SEDIMENTS CAN SOLVE A RANGE OF LOW-BIOAVAILABILITY SITUATIONS BY IMPROVING THEIR MODES OF POLLUTANT ACQUISITION (BIOSURFACTANTS, CHEMOTAXIS, ETC.)
2. HOWEVER, THERE ARE A NUMBER OF ENVIRONMENTAL SITUATIONS, FOR EXAMPLE INVOLVING DESORPTION-RESISTANTCOMPOUNDS, WHERE THE PHYSICAL CONSTRAINTS FOR BIODEGRADATION ARE HARDLY MODIFIABLE BY PHYSIOLOGICAL MEANS
Instituto de Recursos Naturales y Agrobiología de SevillaC.S.I.C.
Grupo “Biodegradación y Biorremediación” (PAI-RNM312)
Marisa BuenoMarta García-Junco*César Gómez*Mohammed Lahlou*Jose Luis NiquiRosa Posada Patricia VelascoJosé Julio Ortega
M. Grifoll (Univ. Barcelona)Hauke Harms (UFZ, Leipzig)Anatoly Marchenko (R. Center Toxicol., Rusia)