Hostname: page-component-cd9895bd7-jn8rn Total loading time: 0 Render date: 2024-12-23T15:17:14.987Z Has data issue: false hasContentIssue false

The amylase activity of 14 species of entodiniomorphid protozoa and the distribution of amylase in rumen digesta fractions of sheep containing no protozoa or one of seven different protozoal populations

Published online by Cambridge University Press:  27 March 2009

G. S. Coleman
Affiliation:
Department of Biochemistry, AFRC Institute of Animal Physiology, Babraham, Cambridge, CB2 4AT

Extract

The amylolytic activity of the cytoplasmic fraction prepared from 14 single species of rumen ciliate protozoa and two natural mixed populations has been determined using five different assays. All the protozoa contained an a-amylase and the major product of amylose digestion was maltose. The highest activities were found in Eremoplastron bovis, Diploplastron affine, Ophryoscolex caudatus and Polyplastron multivesiculatum grown in vitro on grass and wholemeal flour and the lowest in Ostracodinium obtusum bilobum and Diplodinium pentacanthum under all growth conditions and in cultured Entodinium bursa and E. caudatum. The maximum activities depended on the substrate and with Eremoplastron bovis varied from 2·3 (with starch grains) to 48·8 (with amylose) μtmol maltose produced/mg protein per h.

The distribution of amylase between various rumen fractions has been determined at three times after feeding in sheep containing no ciliate protozoa or seven different protozoal populations. A method to give maximal release of the enzyme from mixed rumen bacteria was devised and three different assay methods were compared. The total rumen amylase varied by up to 2·6 times and was highest in animals that contained only Entodinium caudatum or natural A- or B-type populations. The distribution of amylase activity between rumen fractions varied considerably with the various protozoal populations, that in the protozoal cytoplasm being highest in animals containing Epidinium ecaudatum caudatum (46–53%) and the natural A- and B-type populations (35–57 %) and lowest in the animals containing no protozoa (3–11 %) or only Ostra-codinium obtusum bilobum (6–21%). The reverse was true for the activity in the bacterial fraction which contained little activity in animals with natural A- and B-type populations and over 58% of the total activity in an animal containing only Entodinium caudatum. The lowest activities in all fractions were obtained 1·5 h after feeding. Comparatively little activity was found in the plant debris or clarified rumen fluid fractions from any animal. Using results obtained previously the ratios of carboxymethylcellulase to amylase activities have been calculated and shown to be highest in the protozoal cyto-plasmic fraction of Diplodinium pentacanthum and Ostracodinium obtusum bilobum and lowest in Entodinium spp., Diploplastron affine and Eremoplastron bovis.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1986

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Abe, M., Shibui, H., Iriki, T. & Kumeno, F. (1973). Relation between diet and protozoa population in the rumen. Journal of Animal Science 29, 197202.Google Scholar
Abou Akkada, A. R. & Howard, B. H. (1960). The biochemistry of rumen protozoa. 3. The carbohydrate metabolism of Entodinium. Biochemical Journal 76, 445457.CrossRefGoogle Scholar
Bailey, R. W. (1958). Bloat in cattle. X. The carbohydrases of the cattle rumen ciliate Epidinium ecaudatum Crawley isolated from cows fed on red clover (Trifolium pratense L.). New Zealand Journal of Agricultural Research 1, 825833.CrossRefGoogle Scholar
Bailey, R. W. & Clarke, R. T. J. (1963). Carbohydrases of the rumen Oligotrich Eremoplastron bovis. Nature (London) 199, 12911292.Google ScholarPubMed
Coleman, G. S. (1969). The metabolism of starch, maltose, glucose and some other sugars by the rumen ciliate Entodinium caudalum. Journal of General Microbiology 57, 303332.CrossRefGoogle Scholar
Coleman, G. S. (1978). Rumen entodiniomorphtd protozoa. In Methods of Cultivating Parasites in vitro (ed. Baker, J. R. and Taylor, A. E. R.), pp. 3954. London: Academic Press.Google Scholar
Coleman, G. S. (1980). Rumen ciliate protozoa. Advances in Parasitology 18, 121173.CrossRefGoogle ScholarPubMed
Coleman, G. S. (1985). The cellulase content of 15 species of entodiniomorphid protozoa, mixed bacteria and plant debris isolated from the ovine rumen. Journal of Agricultural Science, Cambridge 104, 349360.CrossRefGoogle Scholar
Coleman, G. S. (1986). The distribution of carboxymothylcellulaso between fractions taken from the rumens of sheep containing no protozoa or one of five different protozoal populations. Journal of Agricultural Science, Cambridge 106, 121127.CrossRefGoogle Scholar
Coleman, G. S. & Laurie, J. I. (1974). The metabolism of starch, glucose, amino acids, purines, pyrimidines and bacteria by three Epidinium spp. isolated from the rumen. Journal of General Microbiology 85, 244256.CrossRefGoogle Scholar
Coleman, G. S. & Laurie, J. I. (1977). The metabolism of starch, glucose, amino acids, purines, pyrimidines and bacteria by the rumen ciliate Polyplastron multiveaiculatum. Journal of General Microbiology 98, 679–588.CrossRefGoogle Scholar
Coleman, G. S., Laurie, J. I. & Bailey, J. E. (1977). The cultivation of the rumen ciliate Entodinium bursa in the presence of Entodinium caudatum. Journal of General Microbiology 101, 253258.CrossRefGoogle ScholarPubMed
Coleman, G. S., Laurie, J. I., Bailey, J. E. & Holdgate, S. A. (1976). The cultivation of cellulolytio protozoa isolated from the rumen. Journal of General Microbiology 95, 144150.CrossRefGoogle ScholarPubMed
Coleman, G. S. & Sandford, D. C. (1980). The uptake and metabolism of bacteria, amino acids, glucose and starch by the spined and spineless forms of the rumen ciliate Entodinium caudatum. Journal of General Microbiology 117, 411418.Google ScholarPubMed
Eadie, J. M. (1962). Interrelationships between certain rumen ciliate protozoa. Journal of General Microbiology 29, 579588.CrossRefGoogle Scholar
Eadie, J. M. (1967). Studies on the ecology of certain rumen ciliate protozoa. Journal of General Microbiology 49, 175194.CrossRefGoogle ScholarPubMed
Hnoggrtt, A. St G. & Nixon, D. A. (1957). Use of glucose oxidase, peroxidase and o-dianisidine in determination of blood and urinary glucose. Lancet 2, 368.Google Scholar
Hungate, R. E. (1966). The Rumen and its Microbes. New York: Academic Press.Google Scholar
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265275.CrossRefGoogle ScholarPubMed
Nelson, N. (1944). A photometric adaptation of the Somogyi method for the determination of glucose. Journal of Biological Chemistry 153, 375380.CrossRefGoogle Scholar
Veira, D. M., Ivan, M. & Jui, P. Y. (1983). Rumen ciliate protozoa: effect on digestion in the stomach of sheep. Journal of Dairy Science 66, 10151022.CrossRefGoogle ScholarPubMed
Williams, V. J. & Mckenzie, D. D. S. (1965). The absorption of lactic acid from the reticulo-rumen of the sheep. Australian Journal of Biological Sciences 18, 917934.CrossRefGoogle ScholarPubMed