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A history of in vitro techniques

  • D. J. Minson (a1)


Regressions relating in vivo digestibility to chemical composition of the food have residual standard deviations that are unacceptably high. The development of the two-stage in vitro technique inoculated with rumen liquor (Tilley and Terry, 1963) allowed dry-matter digestibility to be predicted with greater accuracy. This success was followed by a series of developments which replaced rumen liquor with inoculum produced from fresh or preserved faeces collected from sheep or cattle. Other methods used inoculum from a continuous fermentation containing rumen micro-organisms and enzymes produced by fungi. Another modification was to use gas production as a measure of in vitro digestion. The range of nutritional problems that could be measured by in vitro techniques was extended to include the estimation of voluntary food intake and protein degradation. All these in vitro techniques require standardization using food samples that have previously been analysed in vitro or offered as the sole diet to animals. The relative merits of these two calibration methods are discussed. Special facilities are required for storing and distributing these standard foods.



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Burroughs, W., Franks, N. A., Gerlaugh, P. and Beaulieu, Y. 1950. Preliminary observation upon factors influencing cellulose digestion by rumen micro-organisms. Journal of Nutrition 40: 924.
Crocker, B. A., Sniffen, C. J., Hoover, W. H. and Johnson, L. L. 1978. Solvents for soluble nitrogen measurements in feedstuffs. Journal of Dairy Science 61: 437 447.
Czerkawski, J. W. and Breckenridge, G. 1977. Design and development of a long-term rumen simulator technique (Rusitec). British Journal of Nutrition 38: 371384.
Dowman, M. G. and Collins, F. C. 1982. The use of enzymes to predict the digestibility of animal feeds. Journal of the Science of Food and Agriculture 33:689696.
El Shaer, H. M., Omed, H. M., Chamberlain, A. G. and Axford, R. F. E. 1987. Use of faecal organisms from sheep for the in vitro determination of digestibility. Journal of Agricultural Science, Cambridge 109: 257259.
Greenhill, W. L., Couchman, J. F. and Freitas, J. de., 1961. Storage of hay. 3. Effect of temperature and moisture on loss of dry matter and changes in composition. Journal of the Science of Food and Agriculture 12: 293297.
Hungate, R. E. 1966. The rumen and its microbes. Academic Press, New York.
Jarrige, R., Thivend, P. and Demarquilly, C. 1970. Development of a cellulolytic enzyme digestion for predicting the nutritive value of forages. Proceedings of the ninth international grassland congress, Surfers Paradise, Australia, pp. 762766.
Jones, D. I. H. and Hayward, M. V. 1975. The effect of pepsin pretreatment of herbage on the prediction of dry matter digestibility from solubility in fungal cellulase solutions. Journal of the Science of Food and Agriculture 26: 711718.
Kivimae, A. 1960. Estimation of the digestibility of grassland crops from their chemical composition. Proceedings of the eighth international grassland congress, Reading, England,pp. 466470.
Laredo, M. A. and Minson, D. J. 1973. The voluntary intake and digestibility by sheep of leaf and stem fractions of five grasses. Australian Journal of Agricultural Research 24: 875888.
McBee, R. H. 1953. Manometric method for evaluation of microbial activity of rumen with application to utilization of cellulose and hemicellulose. Applied Microbiology 1: 106110.
McLeod, M. N. and Minson, D. J. 1969. Sources of variation in the in vitro digestibility of tropical grasses. Journal of the British Grassland Society 24:244249.
McMeekan, C. P. 1943. A note on the relationship between crude fibre and the digestibility of the organic matter. New Zealand Journal of Science and Technology, A 25: 152153.
Menke, K. H., Raab, L., Salewski, A., Steingass, H., Fritz, D. and Schneider, W. 1979. The estimation of the digestibility and metabolizable energy content of ruminant feedingstuffs from the gas production when they are incubated with rumen liquor in vitro. Journal of Agricultural Science, Cambridge 93: 217222.
Minson, D. J. 1982. Effect of chemical composition on feed digestibility and metabolizable energy. Nutrition Abstracts and Reviews, Series B 52: 591615.
Minson, D. J. 1990. Forage in ruminant nutrition. Academic Press, San Diego.
Minson, D. J. and McLeod, M. N. 1972. The in vitro technique: its modification for estimating digestibility of large numbers of pasture samples. Division of Tropical Crops and Pastures technical paper no 8, pp. 115. CSIRO, Australia.
Owen, E., Jayasuriya, M. C. N., Hamilton, R. and Lalenta, M. 1991. Use of a long-term rumen simulation technique (Rusitec) to provide micro-organisms for in vitro digestibility assays. Journal of Agricultural Science, Cambridge 116: 297301.
Theodorou, M. K. and Brooks, A. E. 1990. Evaluation of a new laboratory procedure for estimating the fermentation kinetics of tropical feeds.Report of the AFRC Institute for Grassland and Environmental Research, Plas Gogerddan, Aberystwyth.
Tilley, J. M. A., , Deriaz, R. E. and Terry, R. A. 1960. The in vitro measurement of herbage digestibility and assessment of nutritive value. Proceedings of the eighth international grassland congress, Reading, England, pp. 533537.
Tilley, J. M. A. and Terry, R. A. 1963. A two stage technique for the in vitro digestion of forage crops. Journal of the British Grassland Society 18:104111.

A history of in vitro techniques

  • D. J. Minson (a1)


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