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Microbial amino acid synthesis and utilization in rats: incorporation of 15N from 15NH4Cl into lysine in the tissues of germ-free and conventional rats

Published online by Cambridge University Press:  09 March 2007

David Torrallardona
Affiliation:
Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB2 9SB
C. Ian Harris
Affiliation:
Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB2 9SB
Marie E. Coates
Affiliation:
School of Biological Sciences, University of Surrey, Guildford, Surrey GU2 5XH
Malcolm F. Fuller
Affiliation:
Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB2 9SB
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Abstract

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The absorption of lysine synthesised by the gastrointestinal microflora was estimated by comparing the15N incorporated into body lysine in four germ-free (15N-GF) and four conventional (15N-CV) rats. They were fed for 10d on a protein-free diet containing fermentable carbohydrates and 15NHM4Cl; another four conventional rats (control), fed on the same diet but with unlabelled NH4Cl, were used to estimate the natural abundance of 15N. The eviscerated carcass of each rat was homogenized and a sample hydrolysed. Lysine was isolated by ion-exchange chromatography and its 15N enrichment was measured by isotope-ratio mass spectrometry. The 15N-CV rats significantly incorporated 15N into their body lysine. The 15N-GF rats had a statistically significant, although small, incorporation of 15body lysine, probably arising from a measurement artifact. It was concluded, therefore, that all [15N]lysine was of microbial origin. The total lysine content in the body and the 15N enrichment of lysine in the microbial fraction of the faeces of the 15N-CV rats were also determined. The amount of microbial lysine absorbed by the 15N-CV rats was estimated by dividing the total amount of [15N]lysine in the body by the enrichment of microbial lysine. It was estimated that the daily absorption of microbial lysine by the conventional rats was 21·3 (SE 2·04) mg/kg body weight0·75

Type
Animal Nutrition
Copyright
Copyright © The Nutrition Society 1996

References

REFERENCES

Åqvist, S. E. G. (1951 a). Metabolic interrelationships among amino acids studied with isotopic nitrogen. Acta Chemica Scandinavica 5, 10461064.CrossRefGoogle Scholar
Åqvist, S. E. G. (1951 b). Amino acid interrelationships during growth, studied with N15-labelled glycine in regenerating rat liver. Acta Chemica Scandinavica 5, 10651073.CrossRefGoogle Scholar
Bender, D. A. (1985). Amino Acid Metabolism. Chichester: John Wiley & Sons.Google Scholar
Bergner, H., Bergner, U. & Adam, K. (1984). Investigations into endogenous N metabolism in 15N labelled pigs. 1. Development of 15N labelling and 15N excretion in urine and faeces after feeding 4 different diets. Archiv für Tierernährung 34, 441455.CrossRefGoogle Scholar
Church, F. C., Porter, D. H., Catignani, G. L. & Swaisgood, H. E. (1985). An o-phthalaldehyde spectro-photometric assay for proteinases. Analytical Biochemistry 146, 343348.CrossRefGoogle Scholar
Coates, M. E. (1984). Diets for germ-free animals. Part 1. Sterilization of diets. In The Germ-Free Animal in Biomedical Research, pp. 8590 [Coates, M. E. and Gustafsson, B. E., editors]. London: Laboratory Animals Ltd.Google Scholar
Coates, M. E. & Gustafsson, B. E. (1984). The Germ-Free Animal in Biomedical Research. London: Laboratory Animals Ltd.Google Scholar
Deguchi, E., Niiyama, M., Kagota, K. & Namioka, S. (1978). Incorporation of 15N administered to germfree and SPF piglets as 15N-urea into amino acids of hydrolyzed liver and muscle protein. Japanese Journal of Veterinary Research 26, 6873.Google Scholar
Deguchi, E., Niiyama, M., Kagota, K. & Namioka, S. (1980). Incorporation of nitrogen-15 from dietary [15N]diammonium citrate into amino acids of liver and muscle proteins in germfree and specific-pathogen-free neonatal pigs. American Journal of Veterinary Research 41, 212214.Google ScholarPubMed
Foster, G. L., Schoenheimer, R. & Rittenberg, D. (1939). Studies in protein metabolism. V. The utilization of ammonia for amino acid and creatine formation in animals. Journal of Biological Chemistry 127, 319327.CrossRefGoogle Scholar
Frape, D. (1986). Equine Nutrition and Feeding. Harlow, Essex: Longman.Google Scholar
Fürst, P. (1972). 15N-studies in severe renal failure. II. Evidence for the essentiality of histidine. Scandinavian Journal of Clinical Laboratory Investigation 30, 307312.CrossRefGoogle Scholar
Giordano, C., de Pascale, C., Balestrieri, C., Cittadini, D. & Crescenzi, A. (1968). The incorporation of urea 15N in amino acids of patients with chronic renal failure on low nitrogen diet. American Journal ofclinical Nutrition 21, 394404.Google ScholarPubMed
Hauck, R. D. (1982). Nitrogen isotope-ratio analysis. In Methods of Soil Analysis, pp. 735779 [Page, A. L., Miller, R. H. and Keeney, D. R., editors]. Madison, WI: American Society of Agronomy Inc.Google Scholar
Heneghan, J. B. (1984). Physiology of the alimentary tract. In The Germ-Free Animal in Biomedical Research, pp. 169191 [Coates, M. E. and Gustafsson, B. E., editors]. London: Laboratory Animals Ltd.Google Scholar
Hentges, D. J. (1992). Gut microflora and disease resistance. In Probiotics, The Scientific Basis, pp. 87110 [Fuller, R., editor]. London: Chapman & Hall.CrossRefGoogle Scholar
Klasing, K. C., Johnstone, B. J. & Benson, B. N. (1991). Implications of an immune response on growth and nutrient requirements of chicks. In Recent Advances in Animal Nutrition, pp. 135146 [Haresign, W. and Cole, D. J. A., editors]. Oxford: Butterworth Heinemann.Google Scholar
Lang, J. (1981). The nutrition of the commercial rabbit. Part I. Physiology, digestibility and nutrient requirements. Nutrition Abstracts and Reviews 51B, 197302.Google Scholar
Laplace, J. P., Darcy Vrillon, B., Duval Iflah, Y. & Raibaud, P. (1985). Proteins in the digesta of the pig; amino acid composition of endogenous, bacterial and fecal fractions. Reproduction, Nutrition, Developpement 25, 10831099.CrossRefGoogle ScholarPubMed
McGinty, D. A., Lewis, H. B. & Marvel, C. S. (1924). Amino acid synthesis in the animal organism. The availability of some caproic acid derivatives for the synthesis of lysine. Journal of Biological Chemistry 62, 7592.CrossRefGoogle Scholar
Markham, R. (1942). A steam distillation apparatus suitable for micro-Kjeldahl analysis. Biochemical Journal 36, 790791.CrossRefGoogle ScholarPubMed
Minitab Inc. (1989). MZNZTAB. State College, PA: Minitab Inc.Google Scholar
National Research Council (1978). Nutrient Requirements of Laboratory Animals. Washington, D.C.: National Academy Press.Google Scholar
Patterson, B. W., Carraro, F. & Wolfe, R. R. (1993). Measurement of 15N enrichment in multiple amino acids and urea in a single analysis by gas chromatography/mass spectrometry. Biological Mass Spectrometry 22, 518523.CrossRefGoogle Scholar
Salter, D. N. (1984). Nitrogen metabolism. In The Germ-Free Animal in Biomedical Research, p. 254 [Coates, M. E. and Gustafsson, B. E., editors]. London: Laboratory Animals Ltd.Google Scholar
Sheng, Y.-B., Badger, T. M., Asplund, J. M. & Wixom, R. L. (1977). Incorporation of 15H4Cl into histidine in adult man. Journal of Nutrition 107, 621630.CrossRefGoogle ScholarPubMed
Spackman, D. H., Stein, W. H. & Moore, S. (1958). Automatic recording apparatus for use in the chromatography of amino acids. Analytical Chemistry 30, 11901206.CrossRefGoogle Scholar
Spedding, F. H., Powell, J. E. & Svec, H. J. (1955). A laboratory method for separating nitrogen isotopes by ion exchange. Journal of the American Chemical Society 77, 61256132.CrossRefGoogle Scholar
Stein, W. H. & Moore, S. (1950). Chromatographic determination of the amino acid composition of proteins. Cold Spring Harbor Symposia on Quantitative Biology, pp. 179190.CrossRefGoogle Scholar
Tanaka, N. (1982). Urea utilization in protein deficient rats. Journal of the Japanese Society of Nutrition and Food Science 35, 175180.Google Scholar
Tanaka, N., Kubo, K., Shiraki, K., Koishi, H. & Yoshimura, H. (1980). A pilot study on protein metabolism in the Papna New Guinea highlanders. Journal of Nutritional Science and Vitaminology 26, 247259.CrossRefGoogle Scholar
Torrallardona, D., Harris, C. I., Milne, E. & Fuller, M. F. (1993 a). Contribution of intestinal microflora to lysine requirements in rats. Proceedings of the Nutrition Society 52, 153A.Google Scholar
Torrallardona, D., Harris, C. I., Milne, E., Ronaasen, V., Coates, M. E. & Fuller, M. F. (1993 b). Contribution of intestinal microflora to lysine requirements in nonruminants. Proceedings of the Nutrition Society 52, 368A.Google Scholar
Torrallardona, D., Harris, C. I. & Fuller, M. F. (1996). Microbial amino acid synthesis and utilization in rats: the role of coprophagy. British Journal of Nutrition 76, 701709.CrossRefGoogle ScholarPubMed
Trexler, P. C. & Reynolds, L. I. (1957). Flexible film apparatus for the rearing and use of germ-free animals. Applied Microbiology 5, 406412.CrossRefGoogle Scholar
Vitti, T. G. & Gaebler, O. H. (1963). Effects of growth hormone on metabolism of nitrogen from several amino acids and ammonia. Archives of Biochemistry and Biophysics 101, 292298.CrossRefGoogle Scholar
Vitti, T. G., Vukmirovich, R. & Gaebler, O. H. (1964). Utilization of ammonia nitrogen, administered by intragastric, intraperitoneal, and subcutaneous routes: effects of growth hormone. Archives of Biochemistry and Biophysics 106, 475482.CrossRefGoogle ScholarPubMed