Main Article Content

Abstract

The increasing significance of aged refuse biofilters for biodegradation processes in wastewater treatment necessitated the constant search for its bacterial properties. This study focused on the diversity of the bacteria present in aged refuse by DNA extraction. The PCR amplifier was used for DNA sequencing using DNA polymerase primer. For PCR amplicons purification, amplicons were extracted from 2% agarose gels and purified using DNA Gel Extraction Kit while quantification was done using QuantiFluor™ -ST. Sample libraries were pooled in equimolar and paired-end sequenced on an Illumina MiSeq platform while Raw fastq files were demultiplexed, quality-filtered using QIIME. The taxonomy of each 16S rRNA gene sequence was evaluated by RDP using confidence threshold of 0.7 while operational Units (OTUs) were clustered with 97% similarity cut-off using UPARSE. The result revealed twelve phyla clusters which include; Firmicutes, Chloroflexi, Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, Planctomycetes, Nitrospirae, Cyanobacteria, Latescibacteria and Gemmatimonadetes. Furthermore, additional group classification revealed up to 513 reads of bacteria kingdom with dominant family classification of Anaerolineaceae, Ruminococcaceae, Nitrosomonadaceae, Planctomycetaceae, Clostridiaceae, Beijerinkiaceae, Xanthobacteracaea, Veillonellacaea, Cemmatimonadacaea. The presence of such diverse organisms in the aged refuse biofilter, show that the aged refuse biofilter has biodegradation potentials when treatment conditions are efficiently optimized.

Keywords

Diversity microbial community aged refuse biofilter Commamox nitrification metagenomics

Article Details

How to Cite
Anijiofor , S. C. ., Che Man, H. ., & Nik Daud, N. N. . (2021). Molecular Diversity and Compositional Analysis of Microbiota in Aged Refuse Biofilter Revealed by Amplicon Sequencing . Basrah Journal of Agricultural Sciences, 34, 138–148. https://doi.org/10.37077/25200860.2021.34.sp1.14

References

  1. Allen, H. K., Moe, L. A., Rodbumrer, J., Gaarder, A., & Handelsman. J., (2009). Functional metagenomics reveals diverse beta-lactamases in a remote Alaskan soil. ISME Journal, 3, 243-251. https://doi.org/10.1038/ismej.2008.86
  2. Anijiofor, S. C., Nor Azreen, M. J., Sarah, J., Saad, S., & Chandima, G. (2017). Aerobic and anaerobic sewage biodegradable processes: The gap analysis. International Journal of Research in Environmental Science, 3(3), 9-19. http://dx.doi.org/10.20431/2454-9444.0303002
  3. Anijiofor, S. C., Nik Daud, N.N., Idrus, S., & Che Man, H. (2018). Recycling of fishpond wastewater by adsorption of pollutants using aged refuse as an alternative low-cost adsorbent. Sustainable Environment Research, 6, 1-7. https://doi.org/10.1016/j.serj.2018.05.005
  4. Cole, J. R., Wang, Q., Cardenas, E., Fish, J., Chai, B., Farris, R. J., Kulam-Syed-Mohideen, A. S., McGarrell, D. M., Marsh, T., Garrity, G. M., & Tiedje, J. M. (2009). The Ribosomal Database Project: improved alignments and new tools for rRNA analysis. Nucleic Acids Research, 37(1), D141-5. https://doi.org/10.1093/nar/gkn879
  5. Cydzik-Kwiatkowska, A., & Zielińska, M. (2016). Bacterial communities in full-scale wastewater treatment systems. World Journal of Microbiology and Biotechnology, 32, 66. https://doi.org/10.1007/s11274-016-2012-9
  6. D’Costa, V. M., McGrann, K. M., Hughes, D. W., & Wright, G. D. (2006). Sampling the antibiotic resistome. Science, 311, 374-377. http://dx.doi.org/ 10.1126/science.1120800
  7. Daims, H., Lebedeva, E. V., Pjevac, P., Han, P., Herbold, C., Albertsen, M., Jehmlich, N., Palatinszky, M., & Vierheilig, J., (2015) Complete nitrification by Nitrospira bacteria. Nature, 528, 504–9. https://doi.org/10.1038/nature16461
  8. DeSantis, T. Z., Hugenholtz, P., Larsen, N., Rojas, M., Brodie, E. L., Keller, K., Huber, T., Dalevi, D., Hu, P., & Andersen, G. L. (2006) Greengenes, a chimera-checked 16s rrna gene database and workbench compatible with ARB. Applied Environmental Microbiology, 72, 5069-5072. https://doi.org/10.1128/AEM.03006-05
  9. Edgar, R.C., Haas, B.J., Clemente, J.C., Quince, C., & Knight, R. (2011). UCHIME improves sensitivity and speed of chimera detection. Bioinformatics, 27 (16), 2194-2200. https://doi.org/ 10.1093/bioinformatics/btr381
  10. Fish, J.A., Chai, B., Wan, Q.; Sun, Y., Titus Brown, C., Tiedje, J.M., Cole, J.R. (2013). FunGene: the functional gene pipeline and repository. Frontiers Microbiology, 4, 291. https://doi.org/10.3389/fmicb.2013.00291
  11. Han, Z., Liu, D., & Liu, Q. B. (2012). A removal mechanism for organics and nitrogen in treating leachate using a semi-aerobic aged refuse biofilter. Journal of Environmental Management, 114, 336-342.https://doi.org/10.1016/j.jenvman.2012.10.026
  12. Jones, R. T., Robeson, M. S., Lauber, C. L., Hamady, M., Knight, R., & Fierer, N. (2009). A comprehensive survey of soil acidobacterial diversity using pyro-sequencing and clone library analyses. ISME Journal, 3(4), 442–453. https://doi.org/10.1038/ismej.2008.127
  13. Justin, J. D., Luke, A. M., Brandon, J. C., Keith, D. S., Flora, C. B., Patricia, S. M., & Jo, H. (2010). Metagenomic analysis of apple orchard soil reveals antibiotic resistance genes encoding predicted bifunctional proteins. Applied and Environmental Microbiology, 76(13), 4396-4401. https://doi.org/10.1128/AEM.01763-09
  14. Koch, H., Lücker, S. Albertsen, M., Kitzinger, K., Herbold, C., Spieck, E., Nielsen, P. H., Wagner, M., & Daims, H., (2015). Expanded metabolic versatility of ubiquitous nitrite-oxidizing bacteria from the genus. Proceedings of the National Academy of Sciences, 112(36), 11371–11376. https://doi.org/10.1073/pnas.1506533112
  15. Kuske, C. R., Ticknor, L. O., Miller, M. E., Dunbar, J. M., Davis, J. A., Barns, S. M., & Belnap, J. (2002). Comparison of soil bacterial communities in rhizospheres of three plant species and the interspaces in an arid grassland. Applied Environmental Microbiology, 68, 1854-1863. https://doi.org/10.1128/AEM.68.4.1854-1863.2002
  16. Liles, M. R., Williamson, L.L., Rodbumrer, J., Torsvik, Goodman, V. R., & Handelsman, J. (2008). Recovery, purification, and cloning of high-molecular-weight DNA from soil microorganisms. Applied Environmental Microbiology, 74, 3302-3305. https://doi.org/10.1128/AEM.02630-07
  17. Lykidis, A., Chen, C.L., Tringe, S.G., McHardy, A.C.,Copeland, A.,Kyrpides, N.C., Hugenholtz, P.,Macarie, H., Olmos, A., Monroy, O., & Liu, W.T. (2010). Multiple syntrophic interactions in a terephthalate-degrading methanogenic consortium. ISME Journal, 5, 122-130. https://doi.org/10.1038/ismej.2010.125
  18. Martín, M.F., & Liras, P. (1989). Organization and expression of genes involved in the biosynthesis of antibiotics and other secondary metabolites. Annual Review of Microbiology, 43, 173-206. https://doi.org/10.1146/annurev.mi.43.100189.00113
  19. McLellan, S. L., Huse, S. M., Mueller-Spitz, S. R., Andreishcheva, E. N., & Sogin, M. L. (2010). Diversity and population structure of sewage-derived microorganisms in wastewater treatment plant influent. Environmental Microbiology, 12, 378-392. https://doi.org/10.1111/j.1462-2920.2009.02075.x
  20. Mohammadi, M., Hassan, M. A., Phang, L.-Y., Shirai, Y., Hasfalina C. M., & Hidayah, A. (2012). Intracellular polyhydroxyalkanoates recovery by cleaner halogen-free method towards zero emission in the palm oil mill. Journal of Cleaner Production, 37, 353-360. https://doi.org/10.1016/j.jclepro.2012.07.038
  21. Muyzer, G., de Waal, E. C., & Uitterlinden, A. G., (1993). Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes encoding for 16S rRNA. Applied Environmental Microbiology, 59, 695-700. https://doi.org/10.1128/aem.59.3.695-700.1993
  22. Pester, M., Maixner, F., Berry, D., Rattei, T., Koch, H., Lücker, S., Nowka, B., Richter, A., & Spieck, E. (2014). NxrB encoding the beta subunit of nitrite oxidoreductase as functional and phylogenetic marker for nitrite- oxidizing Nitrospira. Environmental Microbiology, 16(10), 3055-3071. https://doi.org/10.1111/1462-2920.12300
  23. Prosser, J. I. (2002). Molecular and functional diversity in soil micro-organisms. Plant and Soil, 244, 9-17. https://doi.org/10.1023/A:1020208100281
  24. Quast, C., Pruesse, E., Yilmaz, P., Gerken, J., Schweer, T., Yarza, P., Peplies, J., & Glöckner, F. O. (2013). The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Research, 41(D1), 590-596. https://doi.org/ 10.1093/nar/gks1219
  25. Sogin, M. L., Morrison, H. G., Huber, J. A., Mark, Welch, D., Huse, S. M., Neal, P. R., Arrieta, J. M., & Herndl, G. J. (2006). Microbial diversity in the deep sea and the underexplored ‘‘rare biosphere’’. Proceedings of the National Academy of Sciences of the United States of America, 103, 12115–12120. https://doi.org/10.1073/pnas.0605127103
  26. Van Kessel, M. A. H. J., Speth, D. R., Albertsen, M., Nielsen, P. H., Den Camp O. H. J. M., Kartal, B., Jetten, M. S. M., & Lücker, S. (2015). Complete nitrification by a single microorganism. Nature, 528, 555-559. https://doi.org/10.1038/nature16459
  27. Xie, B., Xiong, S., Liang, S., Hu, C., Zhang, X., & Lu, J. (2012). Performance and bacterial compositions of aged refuse reactors treating mature landfill leachate. Bioresource Technology, 103, 71-77. https://doi.org/10.1016/j.biortech.2011.09.114
  28. Zhang, T., Ye, L., Tong A. H. Y., Shao, M. F., & Lok S., (2011). Ammonia-oxidizing archaea and ammonia-oxidizing bacteria in six full-scale wastewater treatment bioreactors. Applied Microbiology Biotechnology, 91, 1215-1225. https://doi.org/10.1007/s00253-011-3408-y.
  29. Zhao, Y. C., Li, H., Wu, J., & Gu, G. W. (2002). Treatment of leachate by aged-refuse-based biofilter. Journal of Environmental Engineering, 128, 7. https://doi.org/10.1061/(ASCE)0733-9372(2002)128:7(662)

Most read articles by the same author(s)