Abstract
Although our previous studies indicated the two heterotrophic-autotrophic denitrification permeable reactive barriers (HAD PRBs) contained heterotrophic and autotrophic denitrifying bacteria and aerobic heterotrophs, convincing molecular and biochemical evidence for their existence is lacking and the bacterial communities remain largely unknown. Using polymerase chain reaction (PCR) and 16S rRNA, the bacterial community composition in the inoculum introduced into the two HAD PRBs were assessed in this study. The extracted deoxyribonucleic acid (DNA) fragment of about 23 kb in length indicated integral genomic DNA was successfully achieved. The A 260/A 280 ratio of approximately 1.72 suggested the genomic DNA could be directly used for subsequent PCR amplification. The 27F/1492R primer pair was successfully able to obtain an approximately 1500-bp specific band. The inoculum contained aerobic heterotrophic bacteria (belonging to Adhaeribacter and Flavisolibacter), heterotrophic denitrifiers (belonging to Bacillus, Clostridium, Flavobacterium, Steroidobacter and Novosphingobium), hydrogenotrophic denitrifiers (belonging to Pseudomonas) and the other anaerobic bacteria (belonging to Anaerovorax, Azoarcus, Geobacter and Desulfobulbu). The diversity of bacteria from the inoculum was high, with at least 13 bacterial genera present.
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Abbreviations
- BD:
-
Biological denitrification
- DNA:
-
Deoxyribonucleic acid
- dNTP:
-
deoxynucleoside triphosphate
- HAD:
-
Heterotrophic-autotrophic denitrification
- IPTG:
-
Isopropyl-D-thiogalactopyranoside
- LB:
-
Luria–Bertani
- MEGA:
-
Molecular evolutionary genetics analysis
- NCBI:
-
National Center for Biotechnology Information
- PCR:
-
polymerase chain reaction
- PRB:
-
Permeable reactive barrier
- RDP:
-
Ribosomal Database Project
- X-Gal:
-
5-bromo-4-chloro-3-indolyl-D-galactopyranoside
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Liu, F., Huang, G., Fallowfield, H., Guan, H., Zhu, L., Hu, H. (2014). Bacterial Community in the Inoculum. In: Study on Heterotrophic-Autotrophic Denitrification Permeable Reactive Barriers (HAD PRBs) for In Situ Groundwater Remediation. SpringerBriefs in Water Science and Technology. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-38154-6_4
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