ISSN: 0973-7510

E-ISSN: 2581-690X

Research Article | Open Access
Chelladurai Mumtha, Dhanasekaran Subashri and Pambayan Ulagan Mahalingam
Department of Biology, The Gandhigram Rural Institute (Deemed to be University), Gandhigram, Dindigul – 624 302, Tamil Nadu, India.
Article Number: 7727 | © The Author(s). 2022
J Pure Appl Microbiol. 2022;16(3):1861-1872.
Received: 01 April 2022 | Accepted: 23 June 2022 | Published online: 03 August 2022
Issue online: September 2022

This study aims to isolate and characterize efficient biohydrogen generating facultative anaerobic bacteria from various samples, viz., biogas plant (BGP), municipal sewage (MS), and dairy industry treatment plant (DTP). The physicochemical properties of various untreated anaerobic sludge samples reflect the anoxic state and appropriateness of the substrate for separating biohydrogen generating bacteria. The biohydrogen producing bacterial strains were separated from methanogens using the heat-treatment method. The facultative anaerobic bacterial load of heat-treated test samples was determined viz., 27.2±0.57×106 (BGP), 21.8±0.43×106 (MS), and 18.6±0.92×106 (DTP) CFU mL-1 (Colony forming unit), which decreased from the total anaerobic bacterial load of untreated anaerobic sludge viz., 32.1±0.28×106 (BGP), 42.2±0.16×106 (MS), and 34.7±0.12×106 (DTP) CFU mL-1. The 28 predominant bacterial isolates strains were isolated from the heat-treated test samples. All 28 bacterial strains were identified using microscopic and biochemical techniques. Biohydrogen producing potential bacterial strains were screened using the Hungate technique with glucose as a carbon source. Among them, 12 strains were capable of producing biohydrogen, among these 5 strains being excellent biohydrogen producers. Based on the16s rRNA molecular sequencing, the 5 selected biohydrogen generating organisms were authenticated as viz., Salmonella bongori (MZ636759), Escherichia coli (MZ636716), Staphylococcus hominis (MZ636713), Yersinia enterocolitica (OM009292), and Shewanella oneidensis (MZ636800). The gas composition study by GC-TCD in a fermentative medium shows that Shewanella oneidensis (MZ636800) could produce the best biohydrogen (111.4±8.3 mLH2/L), followed by Salmonella bongori (MZ636759) with 98.1±2.9 mL H2/L and Escherichia coli (MZ636716) with 86.7±6.2 mLH2/L.


Biohydrogen, Facultative Anaerobic Bacteria, Anaerobic Sludge, Biogas Plant, Municipal Sewage, Dairy Industry

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© The Author(s) 2022. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.