The mixed samples of contaminated soil, sludge and coke wastewater showed great phenanthrene methanogenic degradation potential.
Comamonadaceae, Nocardiaceae and Methanobacterium were dominant members.
Hexane, hexadecane and benzene could enhance phenanthrene degradation.
Polycyclic aromatic hydrocarbons (PAHs) often occur in oil-contaminated soil, coke wastewater and domestic sludge; however, associated PAH degraders in these environments are not clear. Here we evaluated phenanthrene degradation potential in the mixed samples of above environments, and obtained a methanogenic community with different microbial profile compared to those from sediments. Phenanthrene was efficiently degraded (1.26 mg/L/d) and nonstoichiometric amount of methane was produced simultaneously. 16S rRNA gene sequencing demonstrated that bacterial populations were mainly associated with Comamonadaceae Nocardiaceae and Thermodesulfobiaceae, and that methanogenic archaea groups were dominated by Methanobacterium and Methanothermobacter. Substances such as hexane, hexadecane, benzene and glucose showed the most positive effects on phenanthrene degradation. Substrate utilization tests indicated that this culture could not utilize other PAHs. These analyses could offer us some suggestions on the putative phenanthrene-degrading microbes in such environments, and might help us develop strategies for the removal of PAHs from contaminated soil and sludge.
Fig.1 Phenanthrene degradation rates of zero-order reaction in big flask during different enrichment phases; (phase 1: day 0–5; phase 2: day 7–13; phase 3: day 15?24; phase 4: day 25?39; phase 5: day 45?61; phase 6: day 66?80; phase 7: day 81?101; phase 8: day 105?121; phase 9: day 126?136; phase 10: day 137?155; phase 11: day 158?176; phase 12: day 180?198).
Fig.2 Bacterial members of the initial sample and of the methanogenic enrichment culture; (A) Bacterial groups at class level; (B) Bacterial groups at family level.
Fig.3 Archaeal members at genus level of the initial sample and of the methanogenic enrichment culture; (A) Archaeal groups of initial mixed samples; (B) Archaeal groups of methanogenic enrichment culture.
Fig.4 Relationship of phenanthrene degradation and methane production.
Substrate
degradation capability
Benzene
+
Naphthalene
-
2-methylnaphthalene
-
Phenanthrenecarboxylic acid
+
Phenanthrene
+
Anthracene
?
Fluoranthene
?
Pyrene
?
Tab.1 Substrate utilization tests
Treatment
K
K change
Inoculated control
0.031
Hexane
0.053
67.87%
Hexadecane
0.062
96.60%
Benzene
0.052
65.39%
Phenanthrenecarboxylic acid
0.045
44.62%
Sodium propionate
0.042
34.92%
Sodium acetate
0.031
0.12%
Glucose
0.053
69.29%
Fulvic acid
0.031
?0.23%
Humic acid
0.039
24.18%
AQDS
0.045
42.57%
BES
0.021
?31.53%
Magnetite
0.039
23.22%
SDS
0.049
55.98%
L-rhamnose
0.048
54.34%
Twain-80
0.017
-47.42%
Tab.2 Effects of different substances on phenanthrene degradation rate (K)
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