DOI QR코드

DOI QR Code

Isolation and Characterization of Cellulolytic Anaerobic Fungi from the Guts of the Hanwoo Cattle and the Korean Native Goat

한우 및 산양의 장내 섬유소 분해 혐기 곰팡이의 분리 및 특성 구명

  • 김창현 (한경대학교 동물생명자원학과) ;
  • 이성실 (경상대학교 동물자원과학부)
  • Published : 2003.12.31

Abstract

The study was conducted to isolate and identify highly fibrolytic anaerobic fungi from the guts of a Hanwoo steer and a Korean native goat, and then investigate the characterization of cellulolytic activity of an anaerobic fungus. Twenty-one anaerobic fungal colonies were isolated in the study, in which 16 colonies were isolated from the rumen contents of the Hanwoo steer and 5 colonies from the duodenal fluids of the Korean native goat. Four anaerobic fungi were selected based on higher cellulolytic enzyme activities to identify under a optical microscope. NLRI-M003 and -T004 belong to Neocallimastix genus and NLRI-M014 belongs to Piromyces genus based on the morphology of their thallus, sporangia, rhizoid and the number of flagella. NLRI-M001 appeared to be an unknown strain of anaerobic fungi due to its different morphology from existing types of anaerobic fungi, though the morpholgoy is similar to Orpinomyces sp. Supplementation of 2% anaerobic fungal culture(NLRI-M003) in rumen-mixed microorganisms increased in vitro DM degradability of rice straw and filter paper up to 4 and 11%, respectively, compared with non-supplementation(control). CMCase and xylanase activities in in vitro culture were also higher in 2% fungal supplementation than controls in both rice straw and filter paper substrates.

Keywords

Anaerobic fungi;Rumen;Duodenum;DM degradability;Cellulolytic activity

References

  1. Akin, D. E. and Benner, R. 1988. Degradation of polysaccharides and lignin by ruminal bacteria and fungus. Appl. Environ. Microbiol. 54:1117.
  2. Akin, D. E. 1989. Histrological and physical factors affecting digestibility of forages. Agron. J. 81:17.
  3. Barr, D. J. S. 1980. An outline for the reclassification of the Chytridiales, and for a new order, the Spizellomycetales. Can. J. Bot. 58:2380.
  4. Barr, D. J. S., Kudo, H., Jakober, K. and Cheng, K.-J. 1989. Morphology and development of rumen fungi: Neocallimastix sp, Piromyces communis and Orpinomyces bovis gen. nov., sp. nov. Can. J. Bot. 67:2815.
  5. Bauchop, T. 1979. Rumen anaerobic fungi of cattle and sheep. Appl. Environ. Microbiol. 38:148.
  6. Breton, A., Dusser, M. and Fonty, G. 1989. Morphological and metabolic characterization of a new species of strictly anaerobic rumen fungus; Neocallimastix jouonii. FEMS Microbiol. Lett. 58:309.
  7. Breton, A., Bernalier, A., Dusser, M., Fonty, G., Gaillard-Martinie, B. and Guilot, J. 1990. Anaeromyces mucronatus nov. gen., nov. sp. A new strictly anaerobic rumen fungus with polycentric thallus. FEMS Microbiol. Lett. 70:177.
  8. Breton, A., Dusser, M., Gaillare-Martinie, B., Guillot, J., Millet, L. and Prensier, G. 1991. Piromyces rhizinflata nov. sp., A strictly anaerobic fungus from faeces of the Saharan ass: a morphological, metabolic and ultrastructural study. FEMS Microbiol. Lett. 82:1.
  9. Bryant, M. P. and Burkey, L. A. 1953. Culture methods and some characteristics of some of the mo-re numerous groups of bacteria in the bovine ru-men. J. Dairy Sci. 36:205.
  10. Davies, D. R., Theodorou, M. K., Lawrence, M. I. and Trinci, A. P. J. 1993a. Distribution of anaerobic fungi in the digestive tract of cattle and their survival in faeces. J. Gen. Microbiol. 139:1395.
  11. Davies, D. R., Theodorou, M. K., Brooks, A. E. and Trinci, A. P. J. 1993b. Influence of drying on the survival of anaerobic fungi in rumen digesta and faeces of cattle. FEMS Microbiol. Lett. 106:59.
  12. Gaillard-Martinie, B., Breton, A., Martine, D. and Guillot, J. 1992. Contribution to the morphological and ultrastructural characterization of Piromyces mae, a strictly anaerobic rumen fungus. Current Microbiol. 24:159.
  13. Gold, J. J., Heath, I. B. and Bauchop, T. 1988. Ultrastructural description of new chytrid genus of caecum anaerobe. Caecomyces equi gen. nov. Biosystmes. 21:403.
  14. Gordon, G. L. R. and Phillips, M. W. 1989a. Comparative fermentation properties of anaerobic fungi from the rumen. In 'The Roles of Protozoa and Fungi in Ruminant Digestion' Eds by J. V. Nolan, R. A. Leng and D. I. Demeyer, p. 127. Penambul Books, Armidale, Australia.
  15. Gordon, G. L. R. and Phillips, M. W. 1989b. Degradation and utilization of cellulose and straw by three different anaerobic fungi from the ovine rumen. Appl. Environ. Microbiol. 55:1703.
  16. Heath, I. B., Bauchop, T. and Skipp, R. A. 1983. Assignment of the rumen anaerobe Neocallimastix frontalis to the Spizellomycetales (Chytridiomycetes) on the basis of its polyflagellate zoospore ultrastructure. Can. J. Bot. 61:295.
  17. Ho, Y. W., Barr, D. J. S., Abdullah, N., Jalaludin, S. and Kudo, H. 1993a. Priomyces spiralis, A new species of anaerobic fungus from the rumen of goat. Mycotaxon. XLVII:59.
  18. Ho, Y. W., Barr, D. J. S., Abdullah, N., Jalaludin, S. and Kudo, H. 1993b. Neocallimastix variabilis, A new species of anaerobic fungus from the rumen of cattle. Mycotaxon. XLVI:241.
  19. Ho, Y. W., Bauchop, T., Abdullah, N. and Jalaludin, S. 1990. Ruminomyces elegans gen. et sp. nov. a polycentric anaerobic rumen fungus from cattle. Mycotaxon. 38: 397.
  20. Hungate, R. E. 1966. The rumen and its microbes. Academic Press, Inc., New York, USA.
  21. Joblin, K. N. 1981. NOTES: Isolation enumeration and maintenance of rumen anaerobic fungi in roll tubes. Appl. Environ. Microbiol. 42:1119.
  22. Jones, D. I. H. and Hayward, M. V. 1975. The effect of pepsin pre-treatment of herbage on the prediction of dry matter digestibility from solubility in fungal cellulase solutions. J. Sci. Food Agric. 26:711.
  23. Lee, S. S., Ha, J. K. and Cheng, K.-J., 2000. Influence of an anaerobic fungal culture administration on in vivo ruminal fermentation and nutrient digestion. Anim. Feed Sci. Technol. 88: 201.
  24. Lee, S. S., Moon, Y. H., Kim, C.-H. and Ha, J. K. 2003. In vitro stimulation of rumen microbial fermentation by a rumen anaerobic fungal culture. Anim. Feed Sci. Technol. In press.
  25. Li, J., Heath, I. B. and Bauchop, T. 1990. Piromyces mae and Piromyces dumbonica, two new species of monoflagellated anaerobic chytridiomycete fungi from the hindgut of the horse and elephant. Can. J. Bot. 68: 1021.
  26. Lowe, S. E., Theodorou, M. K., Trinci, A. P. J. and Hespell, R. B. 1985. Growth of anaerobic rumen fungi on defined and semidefined media lacking rumen fluid. J. Gen. Microbiol. 13:2225.
  27. Lowe, S. E., Griffith, G. G., Milne, A., Theodorou, M. K. and Trinci, A. P. J. 1987a. The life cycle and growth kinetics of an anaerobic rumen fungus. J. Gen. Microbiol. 133:1815.
  28. Lowe, S. E., Theodorou, M. K. and Trinci, A. P. J. 1987b. Isolation of anaerobic fungi from saliva and feces of sheep. J. Gen. Microbiol. 133:1829.
  29. Miller, J. L., Blum, R., Glennon, W. E. and Burton, A. L. 1960. Measurement of carboxymethyl cellulase activity. Anal. Biochem. 1:127.
  30. Milne, A., Theodorou, M. K., Jordan, M. G. C., King-Spooner, C. and Trinci, A. P. J. 1989. Survival of anaerobic fungi in faeces, in saliva, and in pure culture. Exp. Mycol. 13:27.
  31. Mountfort, D. O. and Asher, R. A. 1989. Production of xylanase by the ruminal anaerobic fungus Neocallimastix frontalis. Appl. Environ. Microbiol. 55:1016.
  32. Orpin, C. G. 1975. Studies on the rumen flagellate Neocallimastix frontalis. J. Gen. Microbiol. 91:249.
  33. Orpin, C. G. 1976. Studies on the rumen flagellate Sphaeromonas communis. J. Gen. Microbiol. 94:270.
  34. Orpin, C. G. 1977. Invasion of plant tissue in the rumen by the flagellate Neocallimastix frontalis. J. Gen. Microbiol. 98:423.
  35. Orpin, C. G. and Joblin, K. N. 1997. The rumen anaerobic fungi. In 'The Rumen Microbial Ecosystem' Eds by P. N. Hobson and C. S. Stewart, p. 140. St. Edmundsbury Press, Suffolk, UK.
  36. Orpin, C. G. and Munn, E. A. 1986. Neocallimastix patriciarum: new member of the Neocallimsticaccae inhabitating the sheep rumen. Trans. Br. Mycol. Soc. 86: 178.
  37. SAS. 1996. User's Guide: Statistics, Version 6 Editions. SAS Inst., Inc., Cary, NC. USA.
  38. Teunissen, M. J., Op den Camp, H. J. M., Orpin, C. G., Huis in't Veld, J. H. J. and Vogels, G. D. 1991. Comparison of growth characteristics of anaerobic fungi isolated from ruminant and non-ruminant herbivorous during cultivation in a defined medium. J. Gen. Microbiol. 137:1401.
  39. Theodorou, M. K., Gill, M., King-Spooner, C. and Beever, D. E. 1990. Enumeration of anaerobic chytridiomycetes as thallus-forming units: Novel method for quantification of fibrolytic fungal populations from the digestive tract ecosystem. Appl. Environ. Microbiol. 56:1073.
  40. Webb, J. and Theodorou, M. K. 1988. A rumen anaerobic fungus of the genus Neocallimastix: Ultrastructure of the polyflagellate zoospore and young thallus. BioSystems. 21:393.
  41. Williams, A. G. and Orpin, C. G. 1987. Polysaccharide degrading enzymes formed by three species of rumen fungi grown on a range of carbohydrate substrates. Can. J. Microbiol. 33: 418.
  42. Wood, T. M., Wilson, C. A., McCrae, S. I. and Joblin, K. N. 1986. A highly active extracellular cellulase from the anaerobic rumen fungus Neocallimastix frontalis. FEMS Microbiol. Lett. 34:37.
  43. 이성실, 하종규, 최윤재. 1995. 반추위 곰팡이의 분리.동정과 섬유소 분해효소의 특성구명 및 이들의 산업적 이용에 관한 연구. VI. 재래산양에서 분리된 곰팡이의 동정에 관한 연구. 한영사지. 19:152.