Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
권호 표지
DOI QR코드

DOI QR Code

Isolation and characterization of a new Methanoculleus bourgensis strain KOR-2 from the rumen of Holstein steers

  • Battumur, Urantulkhuur (Graduate School of Future Convergence, Hankyong National University) ;
  • Lee, Manhee (Department of Animal Life and Environment Science, General Graduate School, Hankyong National University) ;
  • Bae, Gui Sek (Department of Animal Science and Technology, Chung-Ang University) ;
  • Kim, Chang-Hyun (Department of Animal Life and Environment Science, Hankyong National University)
  • Received : 2018.05.30
  • Accepted : 2018.07.06
  • Published : 2019.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

Objective: To isolate and identify new methanogens from the rumen of Holstein steers in Korea. Methods: Representative rumen contents were obtained from three ruminally cannulated Holstein steers ($793{\pm}8kg$). Pre-reduced media were used for the growth and isolation of methanogens. Optimum growth temperature, pH, and sodium chloride (NaCl) concentration as well as substrate utilization and antibiotic tolerance were investigated to determine the physiological characteristics of the isolated strain. Furthermore, the isolate was microscopically studied for its morphology. Polymerase chain reaction of 16S rRNA and mcrA gene-based amplicons was used for identification. Results: One strain designated as KOR-2 was isolated and found to be a non-motile irregular coccus with a diameter of 0.2 to $0.5{\mu}m$. KOR-2 utilized $H_2+CO_2$ and formate but was unable to metabolize acetate, methanol, trimethylamine, 2-propanol, and isobutanol for growth and methane production. The optimum temperature and pH for the growth of KOR-2 were $38^{\circ}C$ and 6.8 to 7.0, respectively, while the optimum NaCl concentration essential for KOR-2 growth was 1.0% (w/v). KOR-2 tolerated ampicillin, penicillin G, kanamycin, spectromycin, and tetracycline. In contrast, the cell growth was inhibited by chloramphenicol. Phylogenetic analysis of 16S rRNA and mcrA genes revealed the relatedness between KOR-2 and Methanoculleus bourgensis. Conclusion: Based on the physiological and phylogenetic characteristics, KOR-2 was thought to be a new strain within the genus Methanoculleus and named Methanoculleus bourgensis KOR-2.

Keywords

References

  1. Knapp JR, Laur GL, Vadas PA, Weiss WP, Tricarico JM. Invited review: Enteric methane in dairy cattle production: Quantifying the opportunities and impact of reducing emissions. J Dairy Sci 2014;97:3231-61. https://doi.org/10.3168/jds.2013-7234
  2. Johnson KA, Johnson DE. Methane emissions from cattle. J Anim Sci 1995;73:2483-92. https://doi.org/10.2527/1995.7382483x
  3. Patra A, Park T, Kim M, Yu Z. Rumen methanogens and mitigation of methane emission by anti-methanogenic compounds and substances. J Anim Sci Biotechnol 2017;8:13. https://doi.org/10.1186/s40104-017-0145-9
  4. Rother M, Krzycki JA. Selenocysteine, pyrrolysine, and the unique energy metabolism of methanogenic archaea. Archaea 2010;2010:453642. https://doi.org/10.1155/2010/453642
  5. Valle ER, Henderson G, Janssen PH, et al. Considerations in the use of fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy to characterize rumen methanogens and define their spatial distributions. Can J Microbiol 2015;61:417-28. https://doi.org/10.1139/cjm-2014-0873
  6. Battumur U, Yoon YM, Kim CH. Isolation and characterization of a new Methanobacterium formicicum KOR-1 from an anaerobic digester using pig slurry. Asian-Australas J Anim Sci 2016;29:586-93. https://doi.org/10.5713/ajas.15.0507
  7. Battumur U, Yoon YM, Bae GS, Kim CH. Isolation and characterization of new Methanosarcina mazei strains KOR-3, -4, -5, and -6 from an anaerobic digester using pig slurry. Asian-Austalas J Anim Sci 2017;30:1198-205. https://doi.org/10.5713/ajas.16.0830
  8. Ziemer CJ, Sharp R, Stern MD, et al. Comparison of microbial populations in model and natural rumens using 16S ribosomal RNA-targeted probes. Environ Microbiol 2000;2:632-43. https://doi.org/10.1046/j.1462-2920.2000.00146.x
  9. Garrity, GM, Lilburn TG, Cole JR, et al. Taxonomic outline of the Bacteria and Archaea. Part 1. The Archaea, phyla Crenarchaeota and Euryarchaeota [Internet]. Release 7.7. Lansing, MI, USA: Michigan State University; 2007 [cited 2007 Mar 6]. Available from: www.taxonomicoutline.org.
  10. DeLong EF, Pace NR. Environmental diversity of bacteria and archaea. Syst Biol 2001;50:470-8. https://doi.org/10.1080/10635150118513
  11. Janssen PH, Kirs M. Structure of the archaeal community of the rumen. Appl Environ Microbiol 2008;74:3619-25. https://doi.org/10.1128/AEM.02812-07
  12. Lee J-H, Kumar S, Lee G-H, et al. Methanobrevibacter boviskoreani sp. nov., isolated from the rumen of Korean native cattle. Intl J Syst Evol Microbiol 2013;63:4196-201. https://doi.org/10.1099/ijs.0.054056-0
  13. AOAC. Official methods of analysis. 16th ed. Association of Official Analytical Chemists. Washington, DC, USA: AOAC International; 1995.
  14. Van Soest PJ, Robertson JB, Lewis BA. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci 1991;74:3583-97. https://doi.org/10.3168/jds.S0022-0302(91)78551-2
  15. Hungate RE. The anaerobic mesophilic cellulolytic bacteria. Bacteriol Rev 1950;14:1-49. https://doi.org/10.1128/MMBR.14.1.1-49.1950
  16. Balch WE, Fox GE, Magrum LJ, Woese CR, Wolfe RS. Methanogens: reevaluation of a unique biological group. Microbiol Rev 1979;43:260-96. https://doi.org/10.1128/MMBR.43.2.260-296.1979
  17. Sowers KR, Schreier HJ. Media for methanogens. In: Sowers KR, Schreier HJ, editors. Archaea - a laboratory manual methanogens. New York, USA: Cold Spring Harbor Laboratory Press; 1995. p. 459-89.
  18. Sly LI, Blackall LL, Kraat PC, Tian-Shen T, Sangkhobol V. The use of second derivative plots for the determination of mol% guanine plus cytosine of DNA by the thermal denaturation method. J Microbiol Methods 1986;5:139-56. https://doi.org/10.1016/0167-7012(86)90009-6
  19. Grosskopf R, Janssen PH, Liesack W. Diversity and structure of the methanogenic community in anoxic rice paddy soil microcosms as examined by cultivation and direct 16S rRNA gene sequence retrieval. Appl Environ Microbiol 1998;64:960-9. https://doi.org/10.1128/AEM.64.3.960-969.1998
  20. Shlimon AG, Friedrich MW, Niemann H, Ramsing NB, Finster K. Methanobacterium aarhusense sp. nov., a novel methanogen isolated from a marine sediment (Aarhus Bay, Denmark). Int J Syst Evol Micrbiol 2004;54:759-63. https://doi.org/10.1099/ijs.0.02994-0
  21. Luton PE, Wayne JM, Sharp RJ, Riley PW. The mcrA gene as an alternative to 16S rRNA in the phylogenetic analysis of methanogen populations in landfill. Microbiology 2002;148: 3521-30. https://doi.org/10.1099/00221287-148-11-3521
  22. Hales BA, Edwards C, Ritchie DA, et al. Isolation and identification of methanogen-specific DNA from blanket bog feat by PCR amplification and sequence analysis. Appl Environ Microbiol 1996;62:668-75. https://doi.org/10.1128/AEM.62.2.668-675.1996
  23. Simankova MV, Kotsyurbenko OR, Lueders T, et al. Isolation and characterization of new strains of methanogens from cold terrestrial habitats. Syst Appl Microbiol 2003;26:312-8. https://doi.org/10.1078/072320203322346173
  24. Tamura K, Dudley J, Nei M, Kumar S. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 2007;24:1596-9. https://doi.org/10.1093/molbev/msm092
  25. Ollivier BM, Mah RA, Garcia JL, Boone DR. Isolation and characterization of Methanogenium bourgense sp. nov. Int J Syst Bacteriol 1986;36:297-301. https://doi.org/10.1099/00207713-36-2-297
  26. Maestrojuan GM, Boone DR, Xun L, Mah RA, Zhang L. Transfer of Methanogenium bourgens, Methanogenium marisnigri, Methanogenium olentangyi, and Methanogenium thermophilicum to the genus Methanoculleus gen. nov., emendation of Methanoculleus marisnigri and Methanogenium, and description of new strains of Methanoculleus bourgense and Methanoculleus marisnigri. Int J Syst Bacteriol 1990;40:117-22. https://doi.org/10.1099/00207713-40-2-117
  27. Asakawa S, Nagaoka K. Methanoculleus bourgensis, Methanoculleus olentangyi and Methanoculleus oldenburgensis are subjective synonyms. Int J Syst Evol Microbiol 2003;53:1551-2. https://doi.org/10.1099/ijs.0.02508-0
  28. Boone DR, Whitman WB, Koga Y. Family I. Methanomicrobiaceae Barker 1956, 15, AL emend. Balch and Wolfe in Balch, Fox, Magrum, Woese and Wolfe 1979, 286. In: Boone DR, Castenholz RW, Garrity GM, editors. Bergey's manual of systematic bacteriology, vol 1. 2nd ed. The Archaea and the deeply branching and phototrophic Bacteria. New York, USA: Springer; 2001. p. 247.
  29. Oren A. The family Methanomicrobiaceae. In: Rosenberg E, DeLong EF, Lory S, Stackebrandt E, Thompson F, editors. The Prokaryotes: other major lineages of bacteria and the archaea. Berlin, Germany: Springer; 2014. p. 231-46.
  30. Corder RE, Hook LA, Larkin JM, Frea JI. Isolation and characterization of two new methane-producing cocci: Methanogenium olentangyi, sp. nov., and Methanococcus deltae, sp. nov. Arch Microbiol 1983;134:28-32. https://doi.org/10.1007/BF00429402
  31. Blotevogel K-H, Gahl-Janssen R, Jannsen S, et al. Isolation and characterization of a novel mesophilic, fresh-water methanogen from river sediment Methanoculleus oldenburgensis sp. nov. Arch Microbiol 1991;157:54-9. https://doi.org/10.1007/BF00245335
  32. Ferry JG. Methanogenesis: ecology, physiology, biochemistry and genetics. New York, NY, USA: Chapman & Hall; 1993.
  33. Chong SC, Boone DR. Genus II. Methanoculleus Maestrojuan, Boone, Xun, Mah and Zhang 1990, 121VP. In: Boone DR, Castenholz RW, Garrity GM, editors. Bergey's manual of systematic bacteriology, vol 1, 2nd ed. The Archaea and the deeply branching and phototrophic Bacteria. New York, USA: Springer; 2001. p. 251-2.
  34. Cheng L, Qiu TL, Li X, et al. Isolation and characterization of Methanoculleus receptaculi sp. nov. from Shengli oil field, China. FEMS Microbiol Lett 2008;285:65-71. https://doi.org/10.1111/j.1574-6968.2008.01212.x
  35. Lai MC, Chen SC. Methanofollis aquaemaris sp. nov., a methanogen isolated from an aquaculture fish pond. Int J Syst Evol Microbiol 2001;51:1873-80. https://doi.org/10.1099/00207713-51-5-1873
  36. Wu SY, Chen SC, Lai MC. Methanofollis formosanus sp. nov., isolated from a fish pond. Int J Syst Evol Microbiol 2005;55: 837-42. https://doi.org/10.1099/ijs.0.63475-0
  37. Harris JE, Pinn PA, Davis RP. Isolation and characterization of a novel thermophilic, freshwater methanogen. Appl Environ Microbiol 1984;48:1123-8. https://doi.org/10.1128/AEM.48.6.1123-1128.1984
  38. Huber H, Huber G, Stetter KO. Genus VI. Methanoplanus Wildgruber, Thomm and Stetter 1984, 270VP (Effective publication: Wildgruber, Thomm, Konig, Ober, Ricchiuto and Stetter 1982, 36). In: Boone DR, Castenholz RW, Garrity GM, editors. Bergey's manual of systematic bacteriology, vol 1, 2nd ed. The Archaea and the deeply branching and phototrophic Bacteria. New York, USA: Springer; 2001. p. 259-61.
  39. Hilpert R, Winter J, Hammes W, Kandler O. The sensitivity of archaebacteria to antibiotics. Zbl Bakt Mik Hyg I C 1981;2: 11-20.
  40. Luton PE, Wayne JM, Sharp RJ, Riley PW. The mcrA gene as an alternative to 16S rRNA in the phylogenetic analysis of methanogen populations in landfill. Microbiology 2002;148: 3521-30. https://doi.org/10.1099/00221287-148-11-3521

Cited by

  1. Methanogens Diversity during Anaerobic Sewage Sludge Stabilization and the Effect of Temperature vol.8, pp.7, 2019, https://doi.org/10.3390/pr8070822