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Comparison of microbial markers (15N and purine bases) and bacterial isolates for the estimation of rumen microbial protein synthesis

Published online by Cambridge University Press:  18 August 2016

M. D. Carro*
Affiliation:
Departamento de Producción Animal I, Universidad de León, 24071 León, Spain
E. L. Miller
Affiliation:
Department of Clinical Veterinary Medicine, University of Cambridge, Nutrition Laboratory, 307 Huntingdon Road, Cambridge CB3 OJQ, UK
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Abstract

The first objective of this experiment was to investigate the effect of using different bacterial isolates on the estimation of microbial protein production in semi-continuous fermenters (RUSITEC) given four diets, and to test whether a ‘mixed’ bacterial pellet was representative of the whole bacterial population in the fermenters. A second objective was to compare two different microbial markers (nitrogen-fifteen (15N) and purine bases (PB)). Diets consisted of neutral-detergent fibre from grass hay (10 g/day) and sugar-beet pulp (2 g/day) and 280 mg/day of one of four N forms (isolated soya-bean protein, soya-bean peptides, amino acids blended to profile soya-bean protein and NH4Cl). Two 14-day incubation runs were carried out and in each run each of the four different diets were given to two vessels. On days 12 and 13, total digesta (effluent plus nylon bags residues) was collected for analyses of non-ammonia N, 15N enrichment and PB concentration, and for isolation of total mixed bacterial pellets (TB). On the last day of each incubation run, the system was stopped for isolation of liquid- (LAB) and solid-associated (SAB) bacteria. Microbial N flow was estimated from the 15N enrichment and PB concentration in both total digesta and in the three different bacterial pellets (TB, LAB, and SAB). For all diets, LAB presented a greater (P < 005) 15N enrichment and PB: N ratio than SAB, with TB having an intermediate value. For both markers, the use of LAB produced the lowest (P < 005) estimates of microbial N flow and the use of SAB produced the greatest (P < 005) estimates. The use of TB produced intermediate values with all diets, suggesting that TB consisted of SAB and LAB. For all bacterial pellets, PB produced greater (P 005) values of microbial N flow than 15N. However, there was a positive relationship (r = 0·883; P 0001; no. = 15) between the values of microbial N flow determined with the two markers when TB were used as reference.

Type
Research Article
Copyright
Copyright © British Society of Animal Science 2002

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References

Arambel, M. J., Bartley, E. E., Dufva, G. S., Nagaraja, T. G. and Dayton, A. D. 1982. Effect of diet on amino and nucleic acids of rumen bacteria and protozoa. Journal of Dairy Science 65: 20952101.CrossRefGoogle ScholarPubMed
Barrie, S. and Workman, C. T. 1984. An automated analytical system for nutritional investigations using N-15 tracers. Spectroscopy International Journal 3: 439447.Google Scholar
Broderick, G. A. and Merchen, N. R. 1992. Markers for quantifying microbial protein synthesis in the rumen. Journal of Dairy Science 75: 26182632.CrossRefGoogle ScholarPubMed
Calsamiglia, S., Stern, M. D. and Firkins, J. L. 1996. Comparison of nitrogen-15 and purines as microbial markers in continuous culture. Journal of Animal Science 74: 13751381.CrossRefGoogle ScholarPubMed
Carro, M. D. and Miller, E. L. 1999. Effect of supplementing a fibre basal diet with different nitrogen forms on ruminal fermentation and microbial growth in an in vitro semi-continuous culture system (RUSITEC). British Journal of Nutrition 82: 149157.CrossRefGoogle Scholar
Cecava, M. J., Merchen, N. R., Berger, L. L., Mackie, R. I. and Fahey Jr, G. C. 1991. Effects of dietary energy level and protein source on nutrient digestion and ruminal nitrogen metabolism in steers. Journal of Animal Science 69: 22302243.CrossRefGoogle ScholarPubMed
Cecava, M. J., Merchen, N. R., Gay, L. C. and Berger, L. L. 1990. Composition of ruminal bacteria harvested from steers as influenced by dietary energy level, feeding frequency, and isolation techniques. Journal of Dairy Science 73: 24802488.CrossRefGoogle ScholarPubMed
Czerkawski, J. W. and Breckenridge, G. 1977. Design and development of a long-term rumen simulation technique (Rusitec). British Journal of Nutrition 38: 371384.CrossRefGoogle ScholarPubMed
Faichney, G. J. 1980. Measurements in sheep of the quantity and composition of rumen digesta and the fractional outflow rates of digesta constituents. Australian Journal of Agricultural Research 31: 11291137.CrossRefGoogle Scholar
Firkins, J. L., Lewis, S. M., Montgomery, L., Berger, L. L., Merchen, N. R. and Fahey Jr, G. C. 1987. Effects of feed intake and dietary urea concentration on ruminal dilution rate and efficiency of bacterial growth in steers. Journal of Dairy Science 70: 23122321.CrossRefGoogle ScholarPubMed
Komisarczuk, S., Durand, M., Beaumatin, Ph. and Hannequart, G. 1987. Utilisation de l’azote 15 pour la mesure de la protéosynthèse microbienne dans les phases solide et liquide d’un fermenteur semi-continu (Rusitec) (The use of nitrogen-15 for determination of microbial synthesis in the solid and liquid phase of a semi-continuous fermenter (Rusitec)). Reproduction, Nutrition, Développement 27: 261262.CrossRefGoogle Scholar
Legay-Carmier, F. and Bauchart, D. 1989. Distribution of bacteria in the rumen contents of dairy cows given a diet supplemented with soya-bean oil. British Journal of Nutrition 61: 725740.CrossRefGoogle Scholar
Makkar, H. P. S. and Becker, K. 1999. Purine quantification in digesta from ruminants by spectrophotometric and HPLC methods. British Journal of Nutrition 81: 107112.CrossRefGoogle ScholarPubMed
Martin, C., Williams, A. G. and Michalet-Doreau, B. 1994. Isolation and characteristics of the protozoal and bacterial fractions from bovine ruminal contents. Journal of Animal Science 72: 29622968.CrossRefGoogle ScholarPubMed
Martín-Orúe, S. M., Balcells, J., Guada, J. A. and Castrillo, C. 1995. Endogenous purine and pyrimidine derivative excretion in pregnant sows. British Journal of Nutrition 73: 375385.CrossRefGoogle ScholarPubMed
Martín-Orúe, S. M., Balcells, J., Zakraoui, F. and Castrillo, C. 1998. Quantification and chemical composition of mixed bacteria harvested from solid fractions of rumen digesta: effect of detachment procedure. Animal Feed Science and Technology 71: 269282.CrossRefGoogle Scholar
Merry, R. J. and McAllan, A. B. 1983. A comparison of the chemical composition of mixed bacteria harvested from the liquid and solid fractions of rumen digesta. British Journal of Nutrition 50: 701709.CrossRefGoogle ScholarPubMed
Minato, H. and Suto, T. 1978. Technique for fractionation of bacteria in rumen microbial ecosystem. II. Attachment of bacteria isolated from bovine rumen to cellulose powder in vitro and elution of bacteria attached therefrom. Journal of General and Applied Microbiology 24: 116.CrossRefGoogle Scholar
Morrison, M. and Mackie, R. I. 1996. Nitrogen metabolism by ruminal micro-organisms: current understanding and future perspectives. Australian Journal of Agricultural Research 47: 227246.CrossRefGoogle Scholar
Obispo, N. E. and Dehority, B. A. 1999. Feasibility of using total purines as a marker for ruminal bacteria. Journal of Animal Science 77: 30843095.CrossRefGoogle ScholarPubMed
Pérez, J. F., Balcells, J., Fondevilla, M. and Guada, J. A. 1998. Composition of liquid- and particle-associated bacteria and their contribution to the rumen outflow. Australian Journal of Agricultural Research 49: 907914.CrossRefGoogle Scholar
Pérez, J. F., Balcells, J., Guada, J. A. and Castrillo, C. 1997. Rumen microbial production estimated either from urinary purine derivative excretion or from direct measurements of 15N and purine bases as microbial markers: effect of protein source and rumen bacteria isolates. Animal Science 65: 225236.CrossRefGoogle Scholar
Pérez, J. F., Rodriguez, C. A., Gonzalez, J., Balcells, J. and Guada, J. A. 1996. Contribution of dietary purine bases to duodenal digesta in sheep. In situ studies of purine degradability corrected for microbial contamination. Animal Feed Science and Technology 62: 251262.Google Scholar
Rodríguez, C. A., González, J., Alvir, M. R., Repetto, J. L., Centeno, C. and Lamrani, F. 2000. Composition of bacteria harvested from the liquid and solid fractions of the rumen of sheep as influenced by feed intake. British Journal of Nutrition 84: 369376.CrossRefGoogle ScholarPubMed
Smith, R. H., McAllan, A. B., Hewitt, P. and Lewis, P. E. 1978. Estimation of amounts of microbial and dietary nitrogen compounds entering the duodenum of cattle. Journal of Agricultural Science, Cambridge 90: 557568.CrossRefGoogle Scholar
Statistical Analysis Systems Institute. 1989. SAS/STAT® user’s guide (release 6•3). SAS Institute Inc., Cary, NC.Google Scholar
Van Soest, P. J., Robertson, J. B. and Lewis, B. A. 1991. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74: 35833597.CrossRefGoogle ScholarPubMed
Volden, H. 1999. Effects of level of feeding and ruminally undegraded protein on ruminal bacterial protein synthesis, escape of dietary protein, intestinal amino acid profile, and performance of dairy cows. Journal of Animal Science 77: 19051918.CrossRefGoogle ScholarPubMed
Zinn, R. A. and Owens, F. N. 1986. A rapid procedure for purine measurement and its use for estimating net ruminal protein synthesis. Canadian Journal of Animal Science 66: 157166.CrossRefGoogle Scholar