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Methanogenesis in rumen ciliate cultures ofEntodinium caudatum andEpidinium ecaudatum after long-term cultivation in a chemically defined medium

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Abstract

The methanogenic activity in the presence ofEntodinium caudatum andEpidinium ecaudatum was well preserved after long-term cultivation. Microscopic observation revealed that methane production in the presence ofE. caudatum was probably caused by their intracellular methanogenic activity, while methane production in the presence ofE. ecaudatum f.caudatum etecaudatum could be atributed to both the methanogenic bacterial fraction of their external surface and their intracellular activity. Methane production per protozoan cell ofE. caudatum andE. ecaudatum was 2.1 nmol per cell per d and 6.0 nmol. per cell per d, respectively.E. caudatum was responsible for almost the entire methane production in the culture. The activity of free methanogens constituted approximately 50% of the total methane production in thee. ecaudatum culture. Decrease of digestibility of substrates and differences in the fermentation end products accompanied the inhibition of methanogenesis in both cultures by penicillin G. streptomycin, chloramphenicol, 2-bromoethanesulfonate, and pyromellitic diimideE. caudatum appeared to be more sensitive thanE. ecaudatum to the compounds tested. Hydrogen recoveries based on both volatile fatty acids and methane production suggested that the methanogenic population appeared not to be fully able to consume hydrogen produced in the protozoan cultures. The culture conditions tested were found to be suitable for experiments on the relationship between rumen ciliate and rumen bacteria.

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References

  • Cottyn B.G., Boucque C.V.: Rapid method for the gas chromatographic determination of volatile fatty acids in rumen fluid.J. Agric. Chem.16, 105–107 (1968).

    Article  CAS  Google Scholar 

  • Demeyer D.I., Van Nevel C.J.: Methanogenesis, an integrated part of carbohydrate fermentation, and its control, pp. 366–382 inProc. 4th Internat. Symp. Ruminant Physiology. University New England Publ. Unit. Armidale (Australia) 1975.

    Google Scholar 

  • Doddema H.J., Vogels G.D.: Improved identification of methanogenic bacteria by fluorescence microscopy.Appl. Environ. Microbiol.36, 752–756 (1978).

    PubMed  CAS  Google Scholar 

  • Evans V., Bryant J.C., Fioramonti M.C., McQuilkin W.T., Sanford K.K., Earle W.R.: Studies of nutrient media for tissue cellsin vitro. I. A protein-free chemically defined medium for cultivation of strain L-cells.Cancer Res.16, 77–86 (1956).

    PubMed  CAS  Google Scholar 

  • Evans V., Bryant J.C., Kerr H.A., Schilling E.L.: Chemically defined media for cultivation of long-term cell strains from foal mammalian species.Exp. Cell. Res.36, 439–474 (1964).

    Article  PubMed  CAS  Google Scholar 

  • Finlay B.J., Esteban G., Clarke K.J., Williams A.G., Embley T.M., Hirt R.P.: Some rumen ciliates have endosymbiotic methanogens.FEMS Microbiol. Lett.117, 157–162 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Gunsalus R.P., Romesser J.A., Wolfe R.S.: Preparation of coenzyme M analogues and their activity in the methyl-coenzyme M reductase system ofMethanobacterium thermoautotrophicum.Biochemistry16, 2374–2382 (1978).

    Article  Google Scholar 

  • Hillman K., Lloyd D., Williams A.G.: Interactions between the methanogenMethanosarcina barkeri and rumen holotrich ciliate protozoa.Lett. Appl. Microbiol.7, 49–53 (1988).

    Article  Google Scholar 

  • Hino T., Kametaka M., Kandatsu M.: The cultivation of rumen oligotrich protozoa. I. Factors affecting the life ofEntodinium.J. Gen. Appl. Microbiol.19, 305–315 (1973a).

    Article  Google Scholar 

  • Hino T., Kametaka M., Kandatsu M.: The cultivation of rumen oligotrich protozoa. 2. Growth ofEntodinia in vitro.J. Gen. Appl. Microbiol.19, 325–337 (1973b).

    Article  Google Scholar 

  • Hino T., Kametaka M., Kandatsu M.: The cultivation of rumen oligotrich protozoa. 3. White clover factors which stimulate the growth ofEntodinia.J. Gen. Appl. Microbiol.19, 397–413 (1973c).

    Article  CAS  Google Scholar 

  • Itabashi H., Kobayashi T., Matsumoto M.: The effects of rumen ciliate protozoa on energy metabolism and some constituents in rumen fluid and blood plasma of goats.Japan. J. Zootech. Sci.55, 248–256 (1984).

    CAS  Google Scholar 

  • Kišidayova S.: Two-step freezing of the rumen ciliate protozoonEntodinium caudatum.J. Microbiol. Meth.22 185–192 (1995).

    Article  Google Scholar 

  • Kišidayová S.: The cryopreservation of some large ciliate entodiniomorphid protozoa taken from the rumen.Lett. Appl. Microbiol.23, 389–392 (1997).

    Article  Google Scholar 

  • Kišidayová S.: Parameters associated with optimum two-step freezing of rumen ciliatesEntodinium caudatum.J. Microbiol. Meth.30, 119–124 (1997).

    Article  Google Scholar 

  • Kišidayová S.: Cultivation of rumen ciliateEntodinium caudatum in chemicaly defined conditions.J. Eukar. Microbiol.46, 9A-10A (1999).

    Article  Google Scholar 

  • Krumholz L.R., Forsberg C.W.: Association of methanogenic bacteria with rumen protozoa.Can. J. Microbiol.29, 676–680 (1983).

    Article  PubMed  CAS  Google Scholar 

  • Lloyd D., Williams A.G., Amann R., Hayes A.J., Durrant L., Ralphs J.R.: Intracellular prokaryotes in rumen ciliate protozoa: detection by confocal laser scanning microscopy afterin situ hybridization with fluorescent 16S rRNA probes.Eur. J. Protistol.32, 523–531 (1996).

    Google Scholar 

  • Martin S.A., Macy J.M.: Effect of monensin, pyromellitic diimide and 2-bromoethanesulfonic acid on rumen fermentationin vitro.J. Animal. Sci.60, 544–550 (1985).

    CAS  Google Scholar 

  • Menke K.H., Steingass H.: Estimation of energetic feed value obtained from chemical analysis andin vitro gas production using rumen fluid.Animal Res. Dev.28, 7–15 (1989).

    Google Scholar 

  • Michalowski T.: Importance of protein solubility and nature of dietary nitrogen for growth of rumen ciliatesin vitro. pp. 223–231 in J.V. Nolan, R.A. Leng, D.I. Demeyer (Eds):The Roles of Protozoa and Fungi in Ruminant Digestion. Penambul Books. Armidale (Australia) 1989.

    Google Scholar 

  • Muszynski P., Michalowski T., Szcepkowski P.: The influence of protein on the number of rumen ciliatesEntodinium caudatum andDiploplastron affine in vitro.Acta Protozool.24, 307–317 (1985).

    CAS  Google Scholar 

  • Newbold C.J., Lassalas B., Jouany J.P.: The importance of methanogens associated with ciliated with ciliate protozoa in ruminal methane productionin vitro.Lett. Appl. Microbiol.21, 230–234 (1995).

    Article  PubMed  CAS  Google Scholar 

  • Newbold C.J., Ushida K., Morvan B., Fonty G., Jouany J.P.: The role of ciliate protozoa in the lysis of methanogenic archae in rumen fluid.Lett. Appl. Microbiol.23, 421–425 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Quinn L.Y., Burroughs W., Christiansen W.C.: Continuous culture of ruminal microorganisms in chemically defined medium. II. Culture medium studies.Appl. Microbiol.10, 583–592 (1962).

    PubMed  CAS  Google Scholar 

  • Smith M.R.: Reversal of 2-bromoethansulfonate inhibition of metanogenesis inMethanosarcina sp.J. Bacteriol.156, 516–523 (1983).

    PubMed  CAS  Google Scholar 

  • Stumm C.K., Gijzen H.J., Vogels G.D.: Association of methanogenic bacteria with ovine rumen ciliates.Brit. J. Nutr.47, 95–99 (1982).

    Article  PubMed  CAS  Google Scholar 

  • Tokura M., Ushida K., Miyazaki K., Kojima Y.: Methanogens associated with rumen ciliates.FEMS Microbiol. Ecol.22, 137–143 (1997).

    Article  CAS  Google Scholar 

  • Ushida K., Jouany J.I.: Methane production from ciliated rumen protozoa and its effect on protozoan activity.Lett. Appl. Microbiol.23, 129–132 (1996).

    Article  PubMed  CAS  Google Scholar 

  • Ushida K., Jouany J.P., Thivend P.: Role of protozoa in nitrogen digestion in sheep given two isonitrogenous diets.Brit. J. Nutr.56, 407–419 (1986).

    Article  PubMed  CAS  Google Scholar 

  • Ushida K., Newbold C.J., Jouany J.P. Interspecies hydrogen transfer between the rumen ciliatePolyplastron multiversiculatum andMethanosarcina barkeri.J. Gen. Appl. Microbiol.43, 129–131 (1997a).

    Article  PubMed  CAS  Google Scholar 

  • Ushida K., Tokura M., Takenaka A., Itabashi H.: Ciliate protozoa and ruminal methanogenesis, pp. 209–220 in R. Onodera, H. Itabashi, K. Ushida, H. Yano, Y. Sasaki (Eds.):Rumen Microbes and Digestive Physiology in Ruminants. Karger, Basel 1997b.

    Google Scholar 

  • Váradyová Z., Zelenák I., Siroka P.: The comparison ofin vitro fermentation kinetics estimated by three different methods.Arch. Anim Nutr.51, 319–326 (1998).

    Article  Google Scholar 

  • Vogels G.D., Hoppe W.F., Stumm C.K.: Association of methanogenic bacteria with rumen ciliates.Appl. Environ. Microbiol.40, 608–612 (1980).

    PubMed  CAS  Google Scholar 

  • White R.W.: Viable bacteria inside the rumen ciliateEntodinum caudatum.J. Gen. Microbiol.56, 403–408 (1969).

    PubMed  CAS  Google Scholar 

  • Whitelaw F.G., Eadie J.M., Bruce L.A., Shand W.J.: Methane formation in faunated and ciliate-free cattle and its relationship with rumen volatile fatty acid proportions.Brit. J. Nutr.52, 261–275 (1984).

    Article  PubMed  CAS  Google Scholar 

  • Williams A.G., Coleman G.S.:The Rumen Protozoa. Springer-Verlag, New York 1992.

    Google Scholar 

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Authors and Affiliations

  1. Institute of Animal Physiology, Slovak Academy of Sciences, 040 01, Košice, Slovakia

    S. Kišidayová, Z. Váradyová, I. Zeleňák & P. Siroka

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  1. S. Kišidayová
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  2. Z. Váradyová
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  3. I. Zeleňák
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  4. P. Siroka
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Correspondence to S. Kišidayová.

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Kišidayová, S., Váradyová, Z., Zeleňák, I. et al. Methanogenesis in rumen ciliate cultures ofEntodinium caudatum andEpidinium ecaudatum after long-term cultivation in a chemically defined medium. Folia Microbiol 45, 269–274 (2000). https://doi.org/10.1007/BF02908958

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