Isolation and Polyphasic Identification of Hydrolytic Bacterial Communities as a Prospective Bio-remediation Agents for the Waters of River Yamuna
Abstract
Nine isolates of indigenous hydrolytic bacteria with potential as bioremediation agents for the polluted water of river Yamuna, namely Klebsiella pneumoniae, Enterococcus faecium, Aeromonas veronii, Providencia stuartii, Aeromonas veronii, Bacillus sp., E. coli, Bacillus altitudinis, Pseudomonas azotoformans, were obtained by using procedures of isolation, selection and molecular identification by polymerase chain reaction targeting 16s rRNA gene. These bacteria were initially isolated on Nutrient Agar (NA) media and further selection of bacteria was done through hydrolysis tests on Skimmed Milk Agar (SMA), Starch Agar, Tween Agar, CMC and cellulose media as well as hemolysis test for pathogenicity on Blood Agar Plate (BAP). Finally, the isolates were molecularly identified. Bioremediation prospects for river Yamuna have been analyzed.
Keywords:
Bioremediation, Hydrolytic Bacteria, Pollution, River Yamuna
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References
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Cheng, X., Sheng, L., Peng, S., Thorley, E., Cao, H., & Li, K. (2022). Integrated mechanism of heavy metal bioremediation from soil to rice (Oryza sativa L.) mediated by Enterococcus faecium. Plant Growth Regulation, 97(3), 523-535. https://doi.org/10.1007/s10725-022-00811-2
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Deibel, R. H., Lake, D. E., & Niven Jr, C. F. (1963). Physiology of the enterococci as related to their taxonomy. Journal of Bacteriology, 86(6), 1275-1282. https://doi.org/10.1128/jb.86.6.1275-1282.1963
Ekhalil, E. A. I., Gaffar, F. Y., Siddig, M. A. E., & Osman, H. A.H. (2015). Isolation and molecular characterization of cellulolytic Bacillus isolates from soil and compost. American Journal of Microbiological Research, 3(2), 55-58. https://doi.org/10.12691/ajmr-3-2-2
Ethica, S. N., & Raharjo, T. J. (2014). Detection of genes involved in glycerol metabolism of Alcaligenes sp. JG3 (Doctoral dissertation, Universitas Gadjah Mada). https://etd.repository.ugm.ac.id/penelitian/detail/75357
Ethica, S. N., Muchlissin, S. I., Saptaningtyas, R., & Sabdono, A. (2018). Protease producers predominate cultivable hydrolytic bacteria isolated from liquid biomedical waste. Asian Journal of Chemistry, 30(9), 2035-2038. https://doi.org/10.14233/ajchem.2018.21400
Galkiewicz, J. P., & Kellogg, C. A. (2008). Cross-kingdom amplification using bacteria-specific primers: complications for studies of coral microbial ecology. Applied and Environmental Microbiology, 74(24), 7828-7831.https://doi.org/10.1128/AEM.01303-08
Goswami, M., Singh, V., Gusain, P., & Giri, A. (2025). Bioremediation potential of indigenous bacteria species isolated from Yamuna river for the removal of copper and lead. Bioremediation Journal, 1-10. https://doi.org/10.1080/10889868.2025.2504148
Green, M. R., & Sambrook, J. (2012). Molecular cloning. A Laboratory Manual 4th, 448. https://rnajournal.cshlp.org/content/18/7/local/advertising.pdf
Gupta, N., Yadav, K. K., Kumar, V., & Singh, D. (2013). Assessment of physicochemical properties of Yamuna River in Agra City. International Journal of ChemTech Research, 5(1), 528-531. https://www.sphinxsai.com/2013/janmar/chempdf/CT=72(528-531)JM13.pdf
Gupta, P., Samant, K., & Sahu, A. (2012). Isolation of cellulose‐degrading bacteria and determination of their cellulolytic potential. International Journal of Microbiology, 2012(1), 578925. https://doi.org/10.1155/2012/578925
Hashem, J. S., Ismail, W., Chiang, Y. R., Sangal, V., Hentati, D., Abotalib, N., & Bekhit, A. A. (2025). A native bacterial consortium degrades estriol in domestic sewage and activated sludge via the 4, 5-seco pathway and requires estriol to retain its biodegradation phenotype. Microbiology Spectrum, 13(10), e00741-25. https://doi.org/10.1128/spectrum.00741-25
He, T., Zhang, M., Chen, M., Wu, Q., Yang, L., & Yang, L. (2023). Klebsiella oxytoca (EN-B2): A novel type of simultaneous nitrification and denitrification strain for excellent total nitrogen removal during multiple nitrogen pollution wastewater treatment. Bioresource Technology, 367, 128236. https://doi.org/10.1016/j.biortech.2022.128236
Humphrey, C., Underwood, J., Iverson, G., Etheridge, R., O’Driscoll, M., & White, A. (2024). Evaluation of Phosphate and E. coli Attenuation in a Natural Wetland Receiving Drainage from an Urbanized Catchment. Hydrology, 11(6), 74. https://doi.org/10.3390/hydrology11060074
Hussain, A., Rehman, F., Rafeeq, H., Waqas, M., Asghar, A., Afsheen, N., ... & Iqbal, H. M. (2022). In-situ, Ex-situ, and nano-remediation strategies to treat polluted soil, water, and air–A review. Chemosphere, 289, 133252. https://doi.org/10.1016/j.chemosphere.2021.133252
Iqbal, A., Arshad, M., Hashmi, I., Karthikeyan, R., Gentry, T. J., & Schwab, A. P. (2018). Biodegradation of phenol and benzene by endophytic bacterial strains isolated from refinery wastewater-fed Cannabis sativa. Environmental Technology, 39(13), 1705-1714. https://doi.org/10.1080/09593330.2017.1337232
Joshi, P., Chauhan, A., Dua, P., Malik, S., & Liou, Y. (2022). Physicochemical and biological analysis of river Yamuna at Palla station from 2009 to 2019. Scientific Reports, 12(1), 2870. https://doi.org/10.1038/s41598-022-06900-6
Kumar, D., Agrawal, S., & Sahoo, D. (2023). Assessment of the intrinsic bioremediation capacity of a complexly contaminated Yamuna River of India: a algae-specific approach. International Journal of Phytoremediation, 25(13), 1844-1858. https://doi.org/10.1080/15226514.2023.2200862
Kumar, D., Kumar, L., Nagar, S., Raina, C., Parshad, R., & Gupta, V. K. (2012). Screening, isolation and production of lipase/esterase producing Bacillus sp. strain DVL2 and its potential evaluation in esterification and resolution reactions. Archives of Applied Science Research, 4(4), 1763-1770. https://www.scholarsresearchlibrary.com/articles/screening-isolation-and-production-of-lipaseesterase-producing-bacillus-sp-strain-dvl2-and-its-potential-evaluation-in-e.pdf
Kumari, N., & Thakur, S. K. (2014). Randomly amplified polymorphic DNA-a brief review. American Journal of Animal and Veterinary Sciences, 9(1), 6-13. https://doi.org/10.3844/ajavssp.2014.6.13
Mahajan, M., & Prakash, A. (2025). Bacterial Consortia as potential Bioremediation agents for Wastewater Treatment: A Comprehensive Review. International Journal of Advancement in Life Sciences Research, 8(1), 16-33. https://doi.org/https://doi.org/10.31632/ijalsr.2025.v08i01.002
Malik, D., Singh, S., Thakur, J., Singh, R. K., Kaur, A., & Nijhawan, S. (2014). Heavy metal pollution of the Yamuna River: An introspection. International Journal of Current Microbiology and Applied Sciences (IJCMAS), 3(10), 856-863. https://www.ijcmas.com/vol-3-10/Darshan%20Malik,%20et%20al.pdf
McKew, B. A., Coulon, F., Yakimov, M. M., Denaro, R., Genovese, M., Smith, C. J., ... & McGenity, T. J. (2007). Efficacy of intervention strategies for bioremediation of crude oil in marine systems and effects on indigenous hydrocarbonoclastic bacteria. Environmental Microbiology, 9(6), 1562-1571. https://doi.org/10.1111/j.1462-2920.2007.01277.x.
Mishra, T., Tiwari, P. B., Kanchan, S., & Kesheri, M. (2025). Advances in Microbial Bioremediation for Effective Wastewater Treatment. Water, 17(22), 3196. https://doi.org/10.3390/w17223196
Năcescu, N., Israil, A., Cedru, C., & Caplan, D. (1992). Hemolytic properties of some Aeromonas strains. Roumanian Archives of Microbiology and Immunology, 51(3), 147-156. https://www.semanticscholar.org/paper/Hemolytic-properties-of-some-Aeromonas-strains.-N%C4%83cescu-Israil/1dbad4882d6b6fcb1fa25c87d65921cd5852754b
Ozyurek, S. B., & Bilkay, I. S. (2018). Biodegradation of petroleum by Klebsiella pneumoniae isolated from drilling fluid. International Journal of Environmental Science and Technology, 15(10), 2107-2116. https://doi.org/10.1007/s13762-017-1581-y
Paisio, C. E., Quevedo, M. R., Talano, M. A., González, P. S., & Agostini, E. (2014). Application of two bacterial strains for wastewater bioremediation and assessment of phenolics biodegradation. Environmental Technology, 35(14), 1802-1810. https://doi.org/10.1080/09593330.2014.882994
Paisio, C. E., Talano, M. A., González, P. S., Pajuelo-Domínguez, E., & Agostini, E. (2013). Characterization of a phenol-degrading bacterium isolated from an industrial effluent and its potential application for bioremediation. Environmental technology, 34(4), 485-493. https://doi.org/10.1080/09593330.2012.701238
Piyush, P., Kumar, P.S., Madhav, N., V. A., & Mohan, C. (2024). Untreated sewage and industrial effluents in the yamuna river: The impact of urban waste on riverine health and ecosystems. Journal of Emerging Technologies and Innovative Research, 11(10), d763- d767. http://www.jetir.org/papers/JETIR2410382.pdf
Purwaningrum, E., Zulaikhah, S. T., & Ethica, S. N. (2021). Characterization of bacteria from liquid clinical laboratory waste with potential as bioremediation agent. World Journal of Pharmaceutical & Lifa Sciences, 7(9), 16-26. https://www.wjpls.org/home/article_abstract/2351
Ramadhan, A. R., Bachruddin, Z., Erwanto, Y., & Hanim, C. (2021, June). Isolation and selection of proteolytic lactic acid bacteria from colostrum of dairy cattle. In IOP Conference Series: Earth and Environmental Science (Vol. 788, No. 1, p. 012077). IOP Publishing. https://doi.org/10.1088/1755-1315/788/1/012077
Rashid, A., Mirza, S. A., Keating, C., Ali, S., & Campos, L. C. (2022). Indigenous Bacillus paramycoides spp. and Alcaligenes faecalis: sustainable solution for bioremediation of hospital wastewater. Environmental Technology, 43(12), 1903-1916. https://doi.org/10.1080/09593330.2020.1858180
Reza, Z. M., Mohammad, A., Salomeh, K., Reza, A. G., Hossein, S., Maryam, S., ... & Saeed, F. (2014). Rapid detection of coliforms in drinking water of Arak city using multiplex PCR method in comparison with the standard method of culture (Most Probably Number). Asian Pacific Journal of Tropical Biomedicine, 4(5), 404-409. https://doi.org/10.12980/APJTB.4.2014C896
Sharma, A. K., Sharma, V., Saxena, J., Chandra, R., Alam, A., & Prakash, A. (2015). Isolation and screening of amylolytic bacteria from soil. International Journal of Scientific Research in Agricultural Sciences, 2(7), 159-165. http://dx.doi.org/10.12983/ijsras-2015-p0159-0165
Shrivastava, R. K., Shrivastava, S., & Shukla, A. K. (2001). River pollution in India–A brief review. J. Environ. Res, 11(2), 111-115.
Sierra, G. (1957). A simple method for the detection of lipolytic activity of micro-organisms and some observations on the influence of the contact between cells and fatty substrates. Antonie van Leeuwenhoek, 23(1), 15-22. https://doi.org/10.1007/BF02545855
Tantawiwat, S., Tansuphasiri, U., Wongwit, W., Wongchotigul, V., & Kitayaporn, D. (2005). Development of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water. The Southeast Asian Journal of Tropical Medicine and Public Health, 36(1), 162–169. https://www.thaiscience.info/Journals/Article/TMPH/10601144.pdf
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Mahajan, M., & Prakash, A. (2026). Isolation and Polyphasic Identification of Hydrolytic Bacterial Communities as a Prospective Bio-remediation Agents for the Waters of River Yamuna. International Journal of Advancement in Life Sciences Research, 9(1), 219-237. https://doi.org/https://doi.org/10.31632/ijalsr.2026.v09i01.016
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