Czech J. Anim. Sci., 2010, 55(7):276-285 | DOI: 10.17221/74/2009-CJAS
Evaluation of the microbial population in ruminal fluid using real time PCR in steers treated with virginiamycin
- 1 Institute of Animal Science, State Key Laboratory of Animal Nutrition, Chinese Academy of Agricultural Science, Beijing, China
- 2 Xinjiang Agricultural University, Urumqi, China
The objective of the present study was to investigate the effects of virginiamycin (VM ) supplementation on ruminal fermentation and microbial populations in steers. Four ruminally cannulated Chinese Luxi steers (BW 559.4 ± 30.1 kg) were used in a crossover design experiment with an experimental period of 28 days. The forage to concentrate ratio of the basal diet was 35:65 on dry matter basis. The experiment consisted of control treatment and treatment with control diet plus VM at a dose of 30 mg/kg concentrate (DM basis). Rumen fluid was collected at 07:30 prefeeding, at 11:30 and 17:30 postfeeding on day 27 and 28. A part of the pooled sample from rumen fluid was transferred to anaerobic culture by a roll-tube technique and analysed for species-specific real-time PCR quantification. The remaining pooled rumen fluid sample was analyzed for pH, VFA, ammonia N and l-lactic acid. The results showed that VM increased the ruminal pH (6.70 vs. 6.63; P < 0.05), but it decreased ammonia nitrogen (4.94 vs. 6.19 mg/100 ml; P < 0.01) and mean counts of amylolytic bacteria and proteolytic bacteria (P < 0.01) as compared to the control. The additive VM did not affect the l-lactic acid concentration (1.39 vs. 1.26 mmol/l) in rumen fluid. Compared to the control, the steers receiving VM have altered a trend of quantification of Selenomonas ruminantium, Anaerovibrio lipolytica, Ruminococcus albus and Streptococcus bovis in rumen fluid (0.05<p<0.1) as compared to the control. However, VM had no significant effect on Lactobacillus spp. (P = 0.41), Butyrivibrio fibrisolvens (P = 0.35), on the genus Ruminococcus (P = 0.25), Ruminococcus flavefaciens (P = 0.52), Prevotella ruminicola (P = 0.54), on the genus Prevotella (P = 0.67) and Megasphaera elsdenii (P = 0.97). In this study, we found that VM had selective effects on ruminal bacteria and influenced ruminal fermentation by changing a part of the specific ruminal bacteria populations.
Keywords: virginiamycin; roll-tube technique; ruminal fermentation parameters; real-time PCR
Published: July 31, 2010 Show citation
ACS | AIP | APA | ASA | Harvard | Chicago | Chicago Notes | IEEE | ISO690 | MLA | NLM | Turabian | Vancouver |
References
- Boyne A.W., Eadie J.M., Raitt K. (1957): The development and testing of a method of counting rumen ciliate protozoa. Journal of General Microbiology, 17, 414-423.
Go to original source...
Go to PubMed...
- Bramley E., Lean I.J., Fulkerson W.J., Stevenson M.A., Rabiee A.R., Costa N.D. (2008): The definition of acidosis in dairy herds predominantly fed on pasture and concentrates. Journal of Dairy Science, 91, 308-321.
Go to original source...
Go to PubMed...
- Broderick G.A., Kang J.H. (1980): Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media. Journal of Dairy Science, 63, 64-75.
Go to original source...
Go to PubMed...
- Brown M.S., Krehbiel C.R., Galyean M.L., Remmengas M.D., Peters J.P., Hibbard B., Robinson J., Moseley W. M. (2000): Evaluation of models of acute and subacute acidosis on dry matter intake, ruminal fermentation, blood chemistry and endocrine profiles of beef steers. Journal of Animal Science, 78, 3155-3168.
Go to original source...
Go to PubMed...
- Candanosa E., Villa-Godoy A., Castillo D.A., Mendoza G.D. (2008): Effects of monensin, virginiamycin and sodium bicarbonate on ruminal fermentation and acidbase status in sheep. Journal of Animal and Veterinary Advances, 7, 184-189.
- Cheng K.J., Costerton J.W. (1977): Ultrastructure of butyrivibrio fibrisolvens: a gram-positive bacterium. Journal of Bacteriology, 129, 1506-1512.
Go to original source...
Go to PubMed...
- Clayton E.H., Lean I.J., Rowe J.B., Cox J.W. (1999): Effects of feeding virginiamycin and sodium bicarbonate to grazing lactating dairy cows. Journal of Dairy Science, 82, 1545-1554.
Go to original source...
Go to PubMed...
- Coe M.L., Nagaraja T.G., Sun Y.D., Wallace N., Towne E.G., Kemp K.E., Hutcheson J.P. (1999): Effect of virginiamycin on ruminal fermentation in cattle during adaptation to a high concentrate diet and during an induced acidosis. Journal of Animal Science, 77, 2259-2268.
Go to original source...
Go to PubMed...
- Counotte G.H.M., Prins R.A., Janssen R.H.A., DeBie M.J.A. (1981): Role of Megasphaera elsdenii in the fermentation of DL-(2-13C) lactate in the rumen of dairy cattle. Applied and Environmental Microbiology, 42, 649-655.
Go to original source...
Go to PubMed...
- Dehority B.A. (1984): Evaluation of subsampling and fixation procedures used for counting rumen protozoa. Applied and Environmental Microbiology, 48, 182.
Go to original source...
Go to PubMed...
- Dutta G.N., Devriese L.A. (1981): Sensitivity and resistance to growth-promoting agents in animal lactobacilli. Journal of Applied Bacteriology, 51, 283.
Go to original source...
Go to PubMed...
- Garrett E. (1996): Subacute rumen acidosis. Clinical signs and diagnosis in dairy herds. Large Animal Veterinarian, 11, 6-10.
- Godfrey S.I., Boyce M.D., Rowe J.B., Speijers E.J. (1992): Changes within the digestive tract of sheep following engorgement with barley. Australian Journal of Agricultural Research, 44, 1093-1101.
Go to original source...
- Gurtler V., Stanisich V.A. (1996): New approaches to typing and identification of bacteria using the 16S-23S rDNA spacer region. Microbiology, 142, 3-16.
Go to original source...
Go to PubMed...
- Hedde R.D., Armstrong D.G., Parish R.C., Quach R. (1980): Virginiamycin effect on rumen fermentation in cattle. Journal of Animal Science, 51, 366-367.
- Hedde R.D., Shor L., Quach R., Free S.M., Parish R.C., Di Cuollo C.J. (1982): Virginiamycin activity and safety in ruminants. In: Proceedings 2nd European Congress for Veterinary Pharmacology and Toxicology. Toulouse, France.
Go to original source...
- Hedde R.D., Shor L., Quach R. (1983): Virginiamycin activity and safety in ruminants. In: Ruckebush Y., Toutain P., Koritz G.D. (eds.): Veterinary pharmacology and toxicology. MTP Press Ltd., Boston, USA, 768-770.
Go to original source...
- Heilig H.G., Zoetendal E.G., Vaughan E.E., Marteau P., Akkermans A.D., de Vos W.M. (2002): Molecular diversity of Lactobacillus spp. and other lactic acid bacteria in the human intestine as determined by specific amplification of 16S ribosomal DNA. Applied and Environmental Microbiology, 68, 114-123.
Go to original source...
Go to PubMed...
- Hungate R.E. (1969): A roll tube method for cultivation of strict anaerobes. In: Norris J.R., Ribbons D.W. (eds.): Methods in Microbiology. Academic Press, London and New York, 3B, 117-132.
Go to original source...
- Ives S.E., Titgemeyer E.C., Nagaraja T.G., del Barrio A., Bindel D.J., Hollis L.C. (2002): Effects of virginiamycin and monensin plus tylosin on ruminal protein metabolism in steers fed corn-based finishing diets with or without wet corn gluten feed. Journal of Animal Science, 80, 3005-3015.
Go to original source...
Go to PubMed...
- Jalč D., Čertík M. (2005): Effect of microbial oil, monensin and fumarate on rumen fermentation in artificial rumen. Czech Journal of Animal Science, 50, 467-472.
Go to original source...
- Jalč D., Čertík M., Kundríková K., Kubelková P. (2009): Effect of microbial oil and fish oil on rumen fermentation and metabolism of fatty acids in artificial rumen. Czech Journal of Animal Science, 54, 229-237.
Go to original source...
- Klieve A.V., Hennessy D., Ouwerkerk D., Forster R.J., Mackie R.I., Attwood G.T. (2003): Establishing populations of Megasphaera elsdenii YE34 and Butyrivibrio fibrisolvens YE44 in the rumen of cattle fed high grain diets. Journal of Applied Bacteriology, 95, 621-630.
Go to original source...
Go to PubMed...
- Koike S., Yabuki H., Kobayashi Y. (2007): Validation and application of real-time PCR assays for representative rumen bacteria. Journal of Animal Science, 78, 135-141.
Go to original source...
- McDonald C.L., Rowe J.B., Gittins S.P. (1994): Feeds and feeding methods for assembly of sheep before export. Australian Journal of Experimental Agriculture, 34, 589-594.
Go to original source...
- Morris F.E., Branine M.E., Galyean M.L., Hubbert M.E., Freeman A.S., Lofgreen G.P. (1990): Effect of rotating monensin plus tylosin and lasalocid on performance, ruminal fermentation, and site and extent of digestion in feedlot cattle. Journal of Animal Science, 68, 3069-3078.
Go to original source...
Go to PubMed...
- Muir L.A., Barreto A. (1979): Sensitivity of Streptococcus bovis to various antibiotics. Journal of Animal Science, 48, 468.
Go to original source...
Go to PubMed...
- Nagaraja T.G., Taylor M.B. (1987): Susceptibility and resistance of ruminal bacteria to antimicrobial feed additives. Applied and Environmental Microbiology, 53, 1620-1625.
Go to original source...
Go to PubMed...
- Nagaraja T.G., Titgemeyer E.C. (2007): Ruminal acidosis in beef cattle: The current microbiological and nutritional outlook. Journal of Dairy Science, 90, E17-E38.
Go to original source...
Go to PubMed...
- Nagaraja T.G., Taylor M.B., Harmon D.L., Boyer J.E. (1987): In vitro lactic acid inhibition and alterations in volatile fatty acid production by antimicrobial feed additives. Journal of Animal Science, 65, 1064-1076.
Go to original source...
Go to PubMed...
- Nagaraja T.G., Godfrey S.I., Winslow S.W., Rowe J.B. (1995a): Effect of virginiamycin on ruminal fermentation in faunated or ciliate-free sheep overfed with barley grain. Small Ruminant Research, 17, 1-8.
Go to original source...
- Nagaraja T.G., Godfrey S.I., Winslow S.W., Rowe J.B. (1995b): Responses in ciliated protozoa and rumen fermentation in sheep supplemented with barley and virginiamycin. Australian Journal of Agricultural Research, 46, 523-529.
Go to original source...
- Nagaraja T.G., Newbold C.J., Van Nevel C.J., Demeyer D.I. (1997): Manipulation of ruminal fermentation. In: Hobson P.N., Stewart C.S. (eds.): Rumen Microbial Ecosystem. 2nd ed. Blackie Academic and Professional, London, UK, 523-632.
Go to original source...
- Paterson H., Irwin R., Costeron J.W., Cheng K.J. (1975): Ultrastructure and adhesion properties of Ruminococcus albus. Journal of Bacteriology, 122, 278-287.
Go to original source...
Go to PubMed...
- Rogers J.A., Branine M.E., Miller C.R., Wray M.I., Bartle S.J., Preston R.L., Gill D.R., Pritchard R.H., Stillborn R.P., Bechtol D.J. (1995): Effects of dietary virginiamycin on performance and liver abcess incidence in feedlot cattle. Journal of Animal Science, 73, 9-20.
Go to original source...
Go to PubMed...
- Rowe J.B., Pethick D.W. (1994): Starch digestion in ruminants problems, solutions and opportunities. Proceedings of the Nutrition Society of Australia, 18, 40-52.
- Russell J.B., Strobel H.J., Chen G.J. (1988): Enrichment and isolation of a ruminal bacterium with a very high specific activity of ammonia production. Applied and Environmental Microbiology, 54, 872-877.
Go to original source...
Go to PubMed...
- Sambrook J., Fritsch E.F., Maniatis T. (1989): Molecular cloning: a laboratory manual. 2nd ed. In: Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
- Stevenson D.M., Weimer P.J. (2007): Dominance of prevotella and low abundance of classical ruminal bacterial species in the bovine rumen as revealed by relative quantification real-time PCR. Applied Microbiology and Biotechnology, 75, 165-174.
Go to original source...
Go to PubMed...
- Tajima K., Aminov S., Arai T., Nagamine H., Matsui M., Nakamura R.I., Benno Y. (2001): Diet-dependent shift in the bacterial population of the rumen revealed with real-time PCR. Applied and Environmental Microbiology, 67, 2766-2774.
Go to original source...
Go to PubMed...
- Valentine S.C., Clayton E.H., Judson G.J., Rowe J.B. (2000): Effect of virginiamycin and sodium bicarbonate on milk production, milk composition and metabolism of dairy cows fed high levels of concentrates. Australian Journal of Experimental Agriculture, 40, 773-781.
Go to original source...
- Van Nevel C.J., Demeyer D.I. (1990): Effect of antibiotics, a deaminase inhibitor and sarsaponin on nitrogen metabolism of rumen contents in vitro. Animal Feed Science and Technology, 31, 323-348.
Go to original source...
- Van Nevel C.J., Demeyer D.I., Henderickx H.K. (1984): Effect of virginiamycin on carbohydrate and protein metabolism in the rumen in vitro. Archiv für Tierernaehrung, 34, 149-155.
Go to original source...
Go to PubMed...
- Vanzant E.S., Cochran R.C. (1994): Performance and forage utilization by beef cattle receiving increasing amounts of alfalfa hay as a supplement to low-quality, tall grass-prairie forage. Journal of Animal Science, 72, 1059-1067.
Go to original source...
Go to PubMed...
- Váradyová Z., Kišidayov S., Siroka P., Jalč D. (2007): Fatty acid profiles of rumen fluid from sheep fed diets supplemented with various oils and effect on the rumen ciliate population. Czech Journal of Animal Science, 52, 399-406.
Go to original source...
- Yu Z.T., Morrison M. (2004): Improved extraction of PCR-quality community DNA from digesta and fecal samples. Biotechnology, 36, 808-812.
Go to original source...
Go to PubMed...
- Zinn R.A. (1987): Influence of lasalocid and monensin plus tylosin on comparative feeding value of steamflaked versus dry-rolled corn in diets for feedlot cattle. Journal of Animal Science, 65, 256-266.
Go to original source...
Go to PubMed...
This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.