Hostname: page-component-76fb5796d-22dnz Total loading time: 0 Render date: 2024-04-29T08:48:32.525Z Has data issue: false hasContentIssue false

In vitro microbial growth and rumen fermentation of different substrates as affected by the addition of disodium malate

Published online by Cambridge University Press:  09 March 2007

M. L. Tejido
Affiliation:
Departamento de Producción Animal I, Universidad de León, León 24071, Spain
M. J. Ranilla
Affiliation:
Departamento de Producción Animal I, Universidad de León, León 24071, Spain
R. García-Martínez
Affiliation:
Departamento de Producción Animal I, Universidad de León, León 24071, Spain
M. D. Carro*
Affiliation:
Departamento de Producción Animal I, Universidad de León, León 24071, Spain
*
Get access

Abstract

The effects of two concentrations of disodium malate on the in vitro fermentation of three substrates differing in their forage: concentrate ratio (0·8: 0·2, 0·5: 0·5 and 0·2: 0·8; g/g dry matter; low-, medium- and high-concentrate substrates, respectively) by rumen micro-organisms were studied using batch cultures. Rumen contents were collected from four Merino sheep offered lucerne hay ad libitum and supplemented daily with 400 g concentrate. Disodium malate was added to the incubation bottles to achieve final concentrations of 0, 4 and 8 mmol/l malate and 15N was used as a microbial marker. Gas production was measured at regular intervals from 0 to 120 h of incubation to study fermentation kinetics. When gas production values were corrected for gas released from added malate, no effects (P > 0·05) of malate were detected for any of the estimated gas production parameters. In 17-h incubations, the final pH and total volatile fatty acid (VFA) production were increased (P < 0·001) by the addition of malate, but no changes (P > 0·05) were detected in the final amounts of ammonia-N and lactate. When net VFA productions were corrected for the amount of VFA produced from malate fermentation itself, adding malate did not affect (P > 0·05) the production of acetate, propionate and total VFA. Malate reduced methane (CH4) production by proportionately 0·058, 0·013 and 0·054 for the low-, medium- and high-concentrate substrates, respectively. Adding malate to batch cultures increased (P < 0·01) rumen microbial growth (mean values of 16·6, 18·3 and 18·4 mg of microbial N for malate at 0, 4 and 8 mmol/l, respectively), but did not affect (P > 0·05) its efficiency of growth (55·5, 56·7 and 54·3 mg of microbial N per g of organic matter apparently fermented for malate at 0, 4 and 8 mmol/l, respectively). There were no interactions (P > 0·05) malate × substrate for any of the measured variables, and no differences (P > 0·05) in pH, CH4 production and microbial growth were found between malate at 4 and 8 mmol/l. The results indicate that malate had a beneficial effect on in vitro rumen fermentation of substrates by increasing VFA production and microbial growth, and that only subtle differences in the effects of malate were observed between substrates. Most of the observed effects, however, seem to be due to fermentation of malate itself.

Type
Research Article
Copyright
Copyright © British Society of Animal Science 2005

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

Association of Official Analytical Chemists. 1999. Official methods of analysis, 16th edition, fifth revision. AOAC Inc., Arlington, VA.Google Scholar
Caldwell, D. R. and Bryant, M. P. 1966. Medium without rumen fluid for non-selective enumeration and isolation of rumen bacteria. Applied Microbiology 14: 794801.CrossRefGoogle Scholar
Callaway, T. R. and Martin, S. A. 1996. Effects of organic acid and monensin treatment on in vitro mixed ruminal microorganism fermentation of cracked corn. Journal of Animal Science 74: 19821989.CrossRefGoogle ScholarPubMed
Callaway, T. R., Martin, S. A., Wampler, J. L., Hill, N. S. and Hill, G. M. 1997. Malate content of forage varieties commonly fed to cattle. Journal of Dairy Science 80: 16511655.CrossRefGoogle ScholarPubMed
Carro, M. D., López, S., Valdés, C. and Ovejero, F. J. 1999. Effect of DL-malate on mixed ruminal microorganism fermentation using the rumen simulation technique (RUSITEC). Animal Feed Science and Technology 79: 279288.CrossRefGoogle Scholar
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 semicontinuous culture system (RUSITEC). British Journal of Nutrition 82: 149157.CrossRefGoogle Scholar
Carro, M. D. and Miller, E. L. 2002. Comparison of microbial markers (15N and purine bases) and bacterial isolates for the estimation of rumen microbial protein synthesis. Animal Science 75: 315321.CrossRefGoogle Scholar
Carro, M. D. and Ranilla, M. J. 2003. Effect of the addition of malate on in vitro rumen fermentation of cereal grains. British Journal of Nutrition 89: 279288.CrossRefGoogle ScholarPubMed
Carro, M. D., Ranilla, M. J., Giráldez, F. J., Mantecón, A. R. and Balcells, J. 2003. Effect of malate on digestibility and rumen microbial protein synthesis in growing lambs fed a high-concentrate diet. Book of abstracts of the 54th annual meeting of the European Association for Animal Production, no. 9, p. 142. Wageningen Academic Publishers, Wageningen.Google Scholar
Castillo, C., Benedito, J. L., Méndez, J., Pereira, V., López-Alonso, M., Miranda, M. and Hernández, J. 2004. Organic acids as a substitute for monensin in diets for beef cattle. Animal Feed Science and Technology 115: 101116.CrossRefGoogle Scholar
Demeyer, D. I. 1991. Quantitative aspects of microbial metabolism in the rumen and hindgut. In Rumen microbial metabolism and ruminant digestion (ed. Jouany, J. P.), pp. 217237. INRA Editions, Paris.Google Scholar
Demeyer, D. I. and Henderickx, M. K. 1967. Competitive inhibition of in vitro methane production by mixed rumen bacteria. Archives Internationales de Physiology et de Biochemie 75: 157159.Google ScholarPubMed
Dewhurst, R. J., Davies, D. R. and Merry, R. J. 2000. Microbial protein supply from the rumen. Animal Feed Science and Technology 85: 121.CrossRefGoogle Scholar
European Commission. 2003. Regulation (EC) no. 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition. Official Journal of the European Communities L268: 2943.Google Scholar
Goering, M. K. and Van Soest, P. J. 1970. Forage fiber analysis (apparatus, reagents, procedures and some applications). Agricultural handbook, no. 379. Agricultural Research Services, USDA, Washington, DC.Google Scholar
Gómez, J. A., Tejido, M. L. and Carro, M. D. 2005. Influence of disodium malate on microbial growth and fermentation in Rusitec fermenters receiving medium- and high-concentrate diets. British Journal of Nutrition In press.CrossRefGoogle ScholarPubMed
Harmeyer, H., Höller, H., Martens, H. and Grabe, C. von. 1976. Estimate of microbial protein synthesis in vitro by the simultaneous use of three different isotopic markers. In Tracer studies on non-protein nitrogen for ruminants III, pp. 6979. International Atomic Energy Agency, Vienna.Google Scholar
Illg, D. J. and Stern, M. D. 1994. In vitro and in vivo comparisons of diaminopimelic acid and purines for estimating protein synthesis in the rumen. Animal Feed Science and Technology 48: 4955.CrossRefGoogle Scholar
Martin, S. A. 1998. Manipulation of ruminal fermentation with organic acids: a review. Journal of Animal Science 76: 31233132.CrossRefGoogle ScholarPubMed
Martin, S. A. 2004. Effects of DL-malate on in vitro forage fiber digestion by mixed ruminal micro-organisms. Current Microbiology 48: 2731.CrossRefGoogle Scholar
Martin, S. A. and Streeter, M. N. 1995. Effect of malate on in vitro mixed ruminal microorganism fermentation. Journal of Animal Science 73: 21412145.CrossRefGoogle ScholarPubMed
Martin, S. A., Streeter, M. N., Nisbet, D. J., Hill, G. M. and Williams, S. E. 1999. Effects of DL-malate on ruminal metabolism and performance of cattle fed a high-concentrate diet. Journal of Animal Science 77: 10081015.CrossRefGoogle ScholarPubMed
Montaño, M. F., Chai, W., Zinn-Ware, T. E. and Zinn, R. A. 1999. Influence of malic acid supplementation on ruminal pH, lactic acid utilization, and digestive function in steers fed high-concentrate finishing diets. Journal of Animal Science 77: 780784.CrossRefGoogle ScholarPubMed
Nisbet, D. J. and Martin, S. A. 1990. Effect of dicarboxylic acids and Aspergillus oryzae fermentation extract on lactate uptake by the ruminal bacterium Selenomonas ruminantium. Applied and Environmental Microbiology 56: 35153518.CrossRefGoogle ScholarPubMed
Nisbet, D. J. and Martin, S. A. 1991. Effect of a Saccharomyces cerevisiae culture on lactate utilization by the ruminal bacterium Selenomonas ruminantium. Journal of Animal Science 69: 46284633.CrossRefGoogle ScholarPubMed
Nisbet, D. J. and Martin, S. A. 1993. Effects of fumarate, L-malate, and an Aspergillus oryzae fermentation extract on D-lactate utilization by the ruminal bacterium Selenomonas ruminantium. Current Microbiology 26: 133136.CrossRefGoogle Scholar
Ranilla, M. J., Carro, M. D., López, S., Newbold, J. C. and Wallace, J. 2001. Influence of N source on the fermentation of fibre from barley straw and sugarbeet pulp by ruminal micro-organisms in vitro. British Journal of Nutrition 86: 717724.CrossRefGoogle Scholar
Russell, J. B. and Van Soest, P. J. 1984. In vitro ruminal fermentation of organic acids common in forage. Applied and Environmental Microbiology 47: 155159.CrossRefGoogle ScholarPubMed
Statistical Analysis Systems Institute. 1988. SAS/STAT® user's guide (release 6·03). SAS Institute, Cary, NC.Google Scholar
Taylor, K. A. C. C. 1996. A simple colorimetric assay for muramic acid and lactic acid. Applied Biochemistry and Biotechnology 56: 4958.CrossRefGoogle Scholar
Tejido, M. L., Carro, M. D., Ranilla, M. J. and López, S. 2001. In vitro microbial growth as affected by the type of carbohydrate and the source of nitrogen. Proceedings of the British Society of Animal Science, 2001, p. 152.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
Weatherburn, M. W. 1967. Phenol-hypochlorite reaction for determination of ammonia. Analytical Chemistry 39: 971974.CrossRefGoogle Scholar