IDENTIFICATION OF BACTERIA FROM FOOD COURT WASTEWATER BASED ON 16S rRNA GENE SEQUENCES
Main Article Content
Abstract
Environmental safety and public health are greatly influenced by the microbiological elements such as bacterial community, where these organisms can be used as indicators of environmental quality and sources of potential biological hazards. The identification of bacteria in wastewater is an important aspect of environmental microbiology studies. The purpose of this study was to identify the bacterial species in food court wastewater at State University of Surabaya (UNESA) Campus 1. The methods used were exploratory, including bacterial cultivation, DNA extraction, quantitative and qualitative DNA analysis, amplification of 16S rRNA gene fragments, sequencing, and phylogenetic tree reconstruction. From phylogenetic analysis, five test isolates were related to the families Enterobacteriaceae, Aeromonadaceae, Chromatiaceae, and Halomonadaceae. Isolates BU4 and BU5 were most closely related to genus Aeromonas with a similarity of 98.60% and 97.27%, respectively, while isolate BU3 was closely related to genus Klebsiella with a similarity of 96.91%. Meanwhile, isolates BU1 and BU2 cannot yet be identified into a bacterial genus. The results of this study provide important information regarding the prediction of pathogenic bacteria types, which can serve as the basis for recommending further action to prevent disease, improve water quality, and increase the sterility of the food court area.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
Authors who publish with this journal agree to the following terms:
a). Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Attribution-NonCommercial-ShareAlike 4.0 International that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
b). Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
c). Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
References
Abdelsalam, N. R., Hasan, M. E., Javed, T., Rabie, S. M., El-Wakeel, H. E. D. M., Zaitoun, A. F., Abdelsalam, A. Z., Aly, H. M., Ghareeb, R. Y., Hemeida, A. A., & Shah, A. N. (2022). Endorse-ment and phylogenetic analysis of some Fabaceae plants based on DNA barcoding. Molecular Biology Re-ports, 49(6), 5645-5657. https://doi.org/10.1007/s11033-022-07574-z
Akhtar, N., Syakir Ishak, M. I., Bhawani, S. A., & Umar, K. (2021). Various natural and anthropogenic factors responsible for water quality degradation: A review. Water, 13(19), 2660.
Aminu, R. F., Emurotu, M. O., & Mofolorunsho, K. C. (2024). Applications of Molecular Phylogeny in Disease Diagnosis. Int J Res Virol, 1(1), 1-8.
Ammar, E., & Farag, M. (2019). Microbiological Quality and Contamination Sources of Urban Wastewater in Egypt. Environmental Monitoring and Assessment, 191(9), 550. doi:10.1007/s10661-019-7692-5.
Aragaw, T. A. (2021). Functions of various bacteria for specific pollutants degradation and their application in wastewater treatment: a review. International Journal of Environmental Science and Technology, 18, 2063-2076. https://doi.org/10.1007/s13762-020-03022-2
Byappanahalli, M. N., Nevers, M. B., Korajkic, A., Staley, Z. R., & Harwood, V. J. (2012). Enterococci in the environment. Microbiology and Molecular Biology Reviews, 76(4), 685-706. https://doi.org/10.1128/MMBR.00023-12
Chan, S. S., Khoo, K. S., Chew, K. W., Ling, T. C., & Show, P. L. (2022). Recent advances biodegradation and biosorption of organic compounds from wastewater: Microalgae-bacteria consortium-A review. Bioresource Technology, 344, 126159. https://doi.org/10.1016/j.biortech.2021.126159
Cortés-Sánchez, A. D. J., Diaz-Ramírez, M., Rayas-Amor, A. A., Espinosa-Chaurand, L. D., Torres-Ochoa, E., & Salgado-Cruz, M. D. L. P. (2025). Mi-crobiological Hazards in the Food Chain of Fish and Products, a Focus on Klebsiella spp. Veterinary Scienc-es, 12(2), 133. https://doi.org/10.3390/vetsci12020133
Dong, N., Yang, X., Chan, E. W. C., Zhang, R., & Chen, S. (2022). Klebsiella spe-cies: taxonomy, hypervirulence and multidrug resistance. EBioMedicine, 79. https://doi.org/10.1016/j.ebiom.2022.103998
Eslami, H., Shariatifar, A., Rafiee, E., Shiranian, M., Salehi, F., Hosseini, S. S., Eslami, G., Ghanbari, R., & Ebrahimi, A. A. (2019). Decolorization and biodegradation of reactive Red 198 Azo dye by a new Enterococcus faecalis–Klebsiella variicola bacterial consortium isolated from textile wastewater sludge. World Journal of Microbiology and Biotechnology, 35(3), 38.
Feng, P., Weagant, S. D., & Grant, M. A. (2018). Enumeration of Escherichia coli and the Coliform Bacteria. In Bacteriological Analytical Manual. U.S. Food and Drug Administration. Retrieved from FDA BAM.
Fernández-Bravo, A., & Figueras, M. J. (2020). An update on the genus Aeromonas: taxonomy, epidemiology, and pathogenicity. Microorganisms, 8(1), 129. https://doi.org/10.3390/microorganisms8010129
Gerace, E., Mancuso, G., Midiri, A., Poidomani, S., Zummo, S., & Biondo, C. (2022). Recent advances in the use of molecular methods for the di-agnosis of bacterial infections. Patho-gens, 11(6), 663. https://doi.org/10.3390/pathogens11060663
Gogate, P. R., & Pandit, A. B. (2004). A review of imperative technologies for wastewater treatment II: hybrid methods. Advances in environmental research, 8(3-4), 553-597. https://doi.org/10.1016/S1093-0191(03)00031-5
Golnari, M., Bahrami, N., Milanian, Z., Rabbani Khorasgani, M., Asadollahi, M. A., Shafiei, R., & Fatemi, S. S. A. (2024). Isolation and characterization of novel Bacillus strains with superior probiotic potential: Comparative analysis and safety evaluation. Scientific Reports, 14(1), 1457. https://doi.org/10.1038/s41598-024-51823-z
Guerra, M. E. S., Destro, G., Vieira, B., Li-ma, A. S., Ferraz, L. F. C., Hakansson, A. P., Darrieux, M., & Converso, T. R. (2022). Klebsiella pneumoniae biofilms and their role in disease pathogenesis. Frontiers in cel-lular and infection microbiology, 12, 877995. https://doi.org/10.3389/fcimb.2022.877995
Islam, A. K. (2023). Domestic and industrial wastewater generation and its energy recovery potential in Bangladesh. Cleaner Energy Systems, 6, 100092. https://doi.org/10.1016/j.cles.2023.100092
Jamrah, A., Al-Zghoul, T. M., & Darwish, M. M. (2023). A comprehensive review of combined processes for olive mill wastewater treatments. Case Studies in Chemical and Environmental Engineering, 8, 100493. https://doi.org/10.1016/j.cscee.2023.100493
Janda, J. M., & Abbott, S. L. (2021). The changing face of the family Entero-bacteriaceae (Or-der:“Enterobacterales”): New mem-bers, taxonomic issues, geographic expansion, and new diseases and disease syndromes. Clinical microbi-ology reviews, 34(2), 10-1128. doi: https://doi.org/10.1128/cmr.00174-20
Khan, M. A., Sahile, A. A., Jan, R., Asaf, S., Hamayun, M., Imran, M., ... & Lee, I. J. (2021). Halotolerant bacteria miti-gate the effects of salinity stress on soybean growth by regulating sec-ondary metabolites and molecular re-sponses. BMC Plant Biology, 21(1), 176. https://doi.org/10.1186/s12870-021-02937-3
Kumar, S., Chakraborty, D., Tripathi, A., & Ghangrekar, M. M. (2025). Prospect and challenges of production of high value-added products using purple bacteria in integration with wastewater treatment. In Microbial Biotechnology: Integrated Microbial Engineering for B3–Bioenergy, Bioremediation, and Bioproducts (pp. 307-333). Elsevier. https://doi.org/10.1016/B978-0-443-23796-6.00013-4
Kumari, A., Kamaraj, N., Selvaraj, R., & Nanoth, R. (2025). Emerging trends and future outlook on chromium removal in the lab, pilot scale, and industrial wastewater system: an updated review exploring 10 years of research. Environmental Monitoring and Assessment, 197(5), 547. https://doi.org/10.1007/s10661-025-13904-y
Lau, T. T. V., Puah, S. M., Tan, J. A. M. A., Merino, S., Puthucheary, S. D., & Chua, K. H. (2023). Flagellar motility mediates biofilm formation in Aer-omonas dhakensis. Microbial Patho-genesis, 177, 106059. https://doi.org/10.1016/j.micpath.2023.106059
Lee, P. Y., Costumbrado, J., Hsu, C. Y., & Kim, Y. H. (2012). Agarose gel elec-trophoresis for the separation of DNA fragments. Journal of visualized ex-periments: JoVE, (62), 3923. Doi: https://doi.org/10.3791/3923
Moxley, R. A. (2022). Family Enterobacte-riaceae. Veterinary Microbiology, 41-55.
Rahi, P., Mühle, E., Scandola, C., Touak, G., & Clermont, D. (2024). Genome sequence-based identification of En-terobacter strains and description of Enterobacter pasteurii sp. nov. Micro-biology Spectrum, 12(1), e03150-23. https://dx.doi.org/10.1128/spectrum.03150-23
Rawis, L., Mangangka, I. R., & Legrans, R. R. (2022). Analisis kinerja instalansi pengolahan air limbah (ipal) di rumah sakit bhayangkara tingkat iii manado. Tekno, 20(81). Doi: https://doi.org/10.35793/jts.v20i81.42567
Ringler, C., Rosegrant, M. W., & Paisner, M. S. (2000). Irrigation and water resources in Latin America and the Caribbean: Challenges and strategies.
Sadeghi, H., Aslanimehr, M., Nikkhahi, F., Safari, R., Vafaie, M., & Gholamzadeh Khoei, S. (2024). A Systematic Review and Meta-Analysis of Aeromonas-Associated Diarrhea Among Children in Asia. Surgical Infections, 25(7), 538-545. Doi: https://doi.org/10.1089/sur.2024.090
Santoshi, & Kiran, U. V. (2020, December). Microbial Growth in Wastes and Its Health Impact on Rag Pickers. In In-ternational Conference of the Indian Society of Ergonomics (pp. 349-355). Singapore: Springer Nature Singa-pore. Doi: https://doi.org/10.1007/978-981-16-6982-8_31
Sebayang, A. A., Aritonang, C., Silaban, M. O., & Arwita, W. (2025). Analisis Ana-lisis Cemaran Bakteri Coliform pada Air Limbah Domestik dengan Metode MPN. BIO-CONS: Jurnal Biologi dan Konservasi, 7(1). Doi: https://doi.org/10.31537/biocons.v7i1.2294
Selvarajan, R., Sibanda, T., Pandian, J., & Mearns, K. (2021). Taxonomic and functional distribution of bacterial communities in domestic and hospital wastewater system: implications for public and environmental health. Anti-biotics, 10(9), 1059. Doi: https://doi.org/10.3390/antibiotics10091059
Semenov, A. V., & van Overbeek, L. S. (2021). The Role of Microbial Diversi-ty in Wastewater Treatment and Its Management. Waste Management, 119, 267-275. doi:10.1016/j.wasman.2020.08.013.
Singh, G., Singh, A., Singh, P., Shukla, R., Tripathi, S., & Mishra, V. K. (2021). The fate of organic pollutants and their microbial degradation in water bodies. Pollutants and water management: resources, strategies and scarcity, 210-240. https://doi.org/10.1002/9781119693635.ch9
Sophian, A., & Syukur, A. (2021). Analysis of Purity and Concentration of Isolated DNA in Making Raw DNA of Rat Species: Analysis of Purity and Concentration of Isolated DNA in Making Raw DNA of Rat Species. Eruditio: Indonesia Journal of Food and Drug Safety, 1(2), 1-5. Doi: https://doi.org/10.54384/eruditio.v1i2.75
Stackebrandt, E,, & Ebers, J. (2006) Taxonomic parameters revisited: tarnished gold standards. Microbiol Today 33:152–155
Sun, Y., Ren, X., Rene, E. R., Wang, Z., Zhou, L., Zhang, Z., & Wang, Q. (2021). The degradation performance of different microplastics and their effect on microbial community during composting process. Bioresource Technology, 332, 125133.
Sun, R., Zhang, H., Xu, Y., Zhu, H., Yu, X., & Xu, J. (2021). Klebsiella pneumoni-ae-related invasive liver abscess syn-drome complicated by purulent men-ingitis: a review of the literature and description of three cases. BMC In-fectious Diseases, 21(1), 15. Doi: https://doi.org/10.1186/s12879-020-05702-3
Sun, W., Zhang, Y., Zhang, H., Wu, H., Liu, Q., Yang, F., ... & Zhang, W. (2024). Exploitation of Enterobacter hormaechei for biodegradation of multiple plastics. Science of The Total Environment, 907, 167708. https://doi.org/10.1016/j.scitotenv.2023.167708
Tariq, A., & Mushtaq, A. (2023). Untreated wastewater reasons and causes: A review of most affected areas and cit-ies. Int. J. Chem. Biochem. Sci, 23(1), 121-143.
Weinroth, M. D., Belk, A. D., Dean, C., Noyes, N., Dittoe, D. K., Rothrock Jr, M. J., ... & Wells, J. E. (2022). Considerations and best practices in animal science 16S ribosomal RNA gene sequencing microbiome studies. Journal of Animal Science, 100(2), 1-18. https://doi.org/10.1093/jas/skab346
Wheeler, D., & Bhagwat, M. (2008). BLAST QuickStart: example-driven web-based BLAST tutorial (pp. 149-175). Humana Press.
Yang, L., Wu, T., Li, J., & Li, J. (2018). Bac-terial infections in acute-on-chronic liver failure. In Seminars in Liver Dis-ease (Vol. 38, No. 02, pp. 121-133). Thieme Medical Publishers. Doi: https://doi.org/10.1055/s-0038-1657751
Zhu, Y. G., & Chen, D. (2020). Microbial Contamination in Wastewater Treatment Systems: Impact and Mitigation Strategies. Applied Microbiology and Biotechnology, 104(8), 3507-3521. doi:10.1007/s00253-020-10427-6
