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Study on Heterotrophic-Autotrophic Denitrification Permeable Reactive Barriers (HAD PRBs) for In Situ Groundwater Remediation pp 93–103Cite as

Bacterial Community in the Inoculum

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Part of the book series: SpringerBriefs in Water Science and Technology ((BRIEFSWATER))

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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Author information

Authors and Affiliations

  1. China University of Geosciences, 29 Xueyuan Road, 100083, Beijing, People’s Republic of China

    Fei Liu

  2. Beijing Academy of Food Sciences, 70 Yangqiao, 100068, Beijing, People’s Republic of China

    Guoxin Huang

  3. Flinders University, Sturt Road, Adelaide, SA, 5001, Australia

    Howard Fallowfield & Huade Guan

  4. Geological Publishing House, 31 Xueyuan Road, 100083, Beijing, People’s Republic of China

    Lingling Zhu

  5. Institute of Heilongjiang Province, Hydrogeology and Engineering Geology Prospecting, 29 Diantan Street, 150030, Harbin, People’s Republic of China

    Hongyan Hu

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  1. Fei Liu
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  2. Guoxin Huang
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  3. Howard Fallowfield
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  4. Huade Guan
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Correspondence to Fei Liu .

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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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