Comparison of bacterial communities in leachate from decomposing bovine carcasses

  • Yang, Seung Hak (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration) ;
  • Ahn, Hee Kwon (Department of Animal Biosystems Science, Chungnam National University) ;
  • Kim, Bong Soo (Department of Life Science, Hallym University) ;
  • Chang, Sun Sik (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration) ;
  • Chung, Ki Yong (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Eun Mi (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration) ;
  • Ki, Kwang Seok (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration) ;
  • Kwon, Eung Gi (Hanwoo Research Institute, National Institute of Animal Science, Rural Development Administration)
  • Received : 2017.07.26
  • Accepted : 2017.09.15
  • Published : 2017.11.01


Objective: Burial is associated with environmental effects such as the contamination of ground or surface water with biological materials generated during the decomposition process. Therefore, bacterial communities in leachates originating from the decomposing bovine carcasses were investigated. Methods: To understand the process of bovine (Hanwoo) carcass decomposition, we simulated burial using a lab-scale reactor with a volume of $5.15m^3$. Leachate samples from 3 carcasses were collected using a peristaltic pump once a month for a period of 5 months, and bacterial communities in samples were identified by pyrosequencing of the 16S rRNA gene. Results: We obtained a total of 110,442 reads from the triplicate samples of various sampling time points (total of 15 samples), and found that the phylum Firmicutes was dominant at most sampling times. Differences in the bacterial communities at the various time points were observed among the triplicate samples. The bacterial communities sampled at 4 months showed the most different compositions. The genera Pseudomonas and Psychrobacter in the phylum Proteobacteria were dominant in all of the samples obtained after 3 months. Bacillaceae, Clostridium, and Clostridiales were found to be predominant after 4 months in the leachate from one carcass, whereas Planococcaceae was found to be a dominant in samples obtained at the first and second months from the other two carcasses. The results showed that potentially pathogenic microbes such as Clostridium derived from bovine leachate could dominate the soil environment of a burial site. Conclusion: Our results indicated that the composition of bacterial communities in leachates of a decomposing bovine shifted continuously during the experimental period, with significant changes detected after 4 months of burial.


Hanwoo;Decomposition;Leachate;Bacterial Community;Firmicutes;Pyrosequencing


Grant : Cooperative Research Program for Agriculture Science & Technology Development

Supported by : Rural Development Administration


  1. Ahn H. 2010 FMD outbreak in Korea: Government's response to this emergency and important lessons learned. Dearborn, MI, USA: University of Maine; 2012.
  2. NRC. Committee on toxins and pathogens in biosolids applied to land: advancing standards and practices. Washington DC, USA: National Research Council; 2002.
  3. Brown P. BSE: the final resting place. Lancet 1998;351:1146-7.
  4. Nechitaylo TY, Timmis KN, Byzov BA, et al. Fate of prions in soil: degradation of recombinant prion in aqueous extracts from soil and casts of two earthworm species. Soil Biol Biochem 2010;42:1168-71.
  5. Vass AA, Bass WM, Wolt JD, Foss JE, Ammons JT. Time since death determinations of human cadavers using soil solution. J Forensic Sci 1992;37:1236-53.
  6. Gill-King H. Chemical and ultrastructural aspects of decomposition. In: Haglund WD, Sorg MH, editors. Forensic taphonomy: the postmortem fate of human remains. Boca Raton, FL, USA: CRC Press; 1997.
  7. Carter DO, Tibbett M. Microbial decomposition of skeletal muscle tissue (Ovis aries) in a sandy loam soil at different temperatures. Soil Biol Biochem 2006;38:1139-45.
  8. Roling WFM, van Breukelen BM, Braster M, Lin B, van Verseveld HW. Relationships between microbial community structure and hydrochemistry in a landfill leachate-polluted aquifer. Appl Environ Microbiol 2001;67:4619-29.
  9. Acosta-Martinez V, Acosta-Mercado D, Sotomayor-Ramirez D, Cruz-Rodriguez L. Microbial communities and enzymatic activities under different management in semiarid soils. Appl Soil Ecol 2008;38:249-60.
  10. Keener HM, Elwell DL, Monnin JJ. Procedures and equations for sizing of structures and windrows for composting animal mortalities. Appl Eng Agric 2000;16:681-92.
  11. Yang SH, Hong SH, Cho SB, et al. Characterization of microbial community in the leachate associated with the decomposition of entombed pigs. J Microbiol Biotechnol 2012;22:1330-5.
  12. Yang SH, Lim JS, Khan MA, et al. High-throughput nucleotide sequence analysis of diverse bacterial communities in leachates of decomposing pig carcasses. Genet Mol Biol 2015;38:373-80.
  13. Hamady M, Lozupone C, Knight R. Fast UniFrac: facilitating highthroughput phylogenetic analyses of microbial communities including analysis of pyrosequencing and PhyloChip data. ISME J 2010;4:17-27.
  14. Kim BS, Kim JN, Yoon SH, Chun J, Cerniglia CE. Impact of enrofloxacin on the human intestinal microbiota revealed by comparative molecular analysis. Anaerobe 2012;18:310-20.
  15. Jeon YS, Chun J, Kim BS. Identification of household bacterial community and analysis of species shared with human microbiome. Curr Microbiol 2013;67:557-63.
  16. Kim OS, Cho YJ, Lee K, et al. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol 2012;62:716-21.
  17. Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R. UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 2011;27:2194-200.
  18. Schloss PD, Westcott SL, Ryabin T, et al. Introducing mothur: opensource, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 2009;75:7537-41.
  19. Xu J, Chiang HC, Bjursell MK, Gordon JI. Message from a human gut symbiont: sensitivity is a prerequisite for sharing. Trends Microbiol 2004;12:21-8.
  20. Li A, Chu YN, Wang X, et al. A pyrosequencing-based metagenomic study of methane-producing microbial community in solid-state biogas reactor. Biotechnol Biofuels 2013;6:3.
  21. Sanapareddy N, Hamp TJ, Gonzalez LC, et al. Molecular diversity of a North Carolina wastewater treatment plant as revealed by pyrosequencing. Appl Environ Microbiol 2009;75:1688-96.
  22. Poulsen PHB, Al-Soud WA, Bergmark L, et al. Effects of fertilization with urban and agricultural organic wastes in a field trial-prokaryotic diversity investigated by pyrosequencing. Soil Biol Biochem 2013;57:784-93.
  23. Kragelund C, Levantesi C, Borger A, et al. Identity, abundance and ecophysiology of filamentous Chloroflexi species present in activated sludge treatment plants. FEMS Microbiol Ecol 2007;59:671-82.
  24. Harms C, Schleicher A, Collins MD, Andreesen JR. Tissierella creatinophila sp. nov., a gram-positive, anaerobic, non-spore-forming, creatinine-fermenting organism. Int J Syst Bacteriol 1998;48 Pt 3:983-93.
  25. Berge ACB, Glanville TD, Millner PD, Klingborg DJ. Methods and microbial risks associated with composting of animal carcasses in the United States. J Am Vet Med Assoc 2009;234:47-56.
  26. Kelch WJ, Kerr LA, Pringle JK, Rohrbach BW, Whitlock RH. Fatal Clostridium botulinum toxicosis in eleven Holstein cattle fed round bale barley haylage. J Vet Diagn Invest 2000;12:453-5.
  27. Tracy BP, Jones SW, Fast AG, Indurthi DC, Papoutsakis ET. Clostridia: the importance of their exceptional substrate and metabolite diversity for biofuel and biorefinery applications. Curr Opin Biotechnol 2012;23:364-81.
  28. Stahl DA, Tiedje JM. Microbial ecology and genomics: a crossroads of opportunity. American Academy of Microbiology; 2002. pp. 5-12.
  29. Bowman JP, The genus Psychrobacter. In: Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E, editors. The prokaryotes - a handbook on the biology of bacteria. New York, USA: Springer; 2006. pp. 920-30.

Cited by

  1. with malodour in bloodhound dogs, and the effects of a topical product composed of essential oils and plant-derived essential fatty acids in a randomized, blinded, placebo-controlled study pp.09594493, 2018,