Rumen microbial diversity under influence of a polyclonal antibody preparation against lactate-producing and proteolytic bacteria in cows fed different energy sources

Diversidade microbiana ruminal sob efeito de um preparado de anticorpos policlonais contra bactérias proteolíticas e produtoras de lactato em vacas alimentadas com três diferentes fontes energéticas

Abstracts

Nine ruminally cannulated cows fed different energy sources were used to evaluate an avianderived polyclonal antibody preparation against specific ruminal bacteria and monensin on microbial community diversity. The experimental design was three Latin squares 3 x 3 distinguished by the main energy source in the diet [dry-ground corn grain, high moisture corn silage or citrus pulp]. Inside each Latin square, animals received one of the feed additives per period [control, monensin or polyclonal antibody preparation]. Each period lasted 21 days where 20 were used for treatments adaptation and the last one for sampling collection. Microbial diversity was evaluated by protozoa counts and denaturing gradient gel electrophoresis. Polyclonal antibodies plus citrus pulp (CiPu) addition in the diet resulted in an increase of relative counting of Isotricha protozoa that indicates a possible effect on this ruminal ciliate population. In general lines, in the present experiment, it was not possible to assign that there was a pattern in the structures of amplification of Bacteria and Archaea communities of the ruminal content. Oral passive immunization is a technology that arises as an effective alternative for feed additive production. Further research is still necessary to better understand its mechanisms of action.

denaturing gradient gel by electrophoresis; feed additive; ionophores; passive immunity; protozoa


Nove fêmeas bovinas canuladas no rúmen alimentadas com diferentes fontes energéticas foram utilizadas para avaliar um preparado de anticorpos policlonais aviário contra bactérias ruminais específicas e monensina na diversidade da comunidade microbiana. O design experimental foi três quadrados latinos 3 x 3 distintos pela principal fonte energética na dieta [milho seco e moído, silagem de grão úmido de milho ou polpa cítrica]. Dentro de cada quadrado latino, os animais receberam um dos aditivos alimentares [controle, monensina ou preparado de anticorpos policlonais]. Cada período teve duração de 21 dias onde 20 dias foram utilizados para adaptação aos tratamentos e o último dia para colheita das amostras. A diversidade microbiana foi avaliada pela contagem de protozoários e pela técnica da eletroforese em gel com gradiente desnaturante. Anticorpos policlonais adicionado a polpa cítrica na dieta resultou em um aumento na contagem relativa de Isotricha o que pode indicar um efeito sobre esta população de ciliado ruminal. Em linhas gerais, no presente estudo, não foi possível determinar um padrão nas estruturas de amplificação das comunidades Bacteria e Archaea do conteúdo ruminal. A imunização passiva oral é uma tecnologia que surge como alternativa eficiente para a produção de aditivos alimentares. Novas pesquisas são ainda necessárias para melhor compreender seu mecanismo de ação.

aditivo alimentar; eletroforese em gel com gradiente desnaturante; imunidade passiva; ionóforos; protozoário


  • BASTOS, J.P.S.T. Effect of different doses of specific polyclonal antibody preparation on ruminal variables, in situ degradability and in vivo digestibility in cattle fed high concentrate diets 2009. 111p. Diss (MSc). São Paulo State University, Botucatu, Brazil.
  • BERGHMAN, L.R.; WAGHELA, S.D. Antibodies: an alternative for antibiotics? Journal of Animal Science, v.82, p.660-666, 2004.
  • BERGMAN, E. N. Energy contributions of volatile fatty acids from the gastrointestinal tract in various species. Physiological Reviews, v.70, n.2, p.567-590, 1990.
  • BLANCH, M.; CALSAMIGLIA, S.; DiLORENZO, N.; DiCOSTANZO, A.; MUETZEL, S.; WALLACE, R. J. Physiological changes in rumen fermentation during acidosis induction and its control using a multivalent polyclonal antibody preparation in heifers. Journal of Animal Science, v.87, n.5, p.1722-1730, 2009.
  • CARVALHO, L.F.P.B.; AMORIM, G.L.; MATOS, D.S.D.; BATISTA, A.M.V.; MORAES, A.C.A.D.; CABRAL, A.M.D. Protozoários do rúmen de caprinos submetidos a dieta com casca de soja. Revista Brasileira de Sáude e Produção Animal [Online], v.12, n.1, p.244-253, 2011.
  • DEHORITY, B.A. Laboratory manual for classification and morphology of rumen ciliate protozoa Florida, USA: CRC Press Inc., 1993.
  • DEHORITY, B.A. Effect of pH on viability of Entodinium caudatum, Entodinium exiguum, Epidinium caudatum and Ophryoscolex purkynjei in vitro. The Journal of Eukaryotic Microbiology, v.52, n.4, p.339-342, 2005.
  • DiLORENZO, N.; DIEZ-GONZALEZ, F.; DiCONSTANZO, A. Effects of feeding polyclonal antibodies preparations on ruminal bacterial populations and ruminal pH of steers fed high-grain diets. Journal of Animal Science, v.84, n.8, p.2178-2185, 2006.
  • DiLORENZO, N.; DAHLEN, C.R.; DIEZ-GONZALEZ, F.; LAMB, G.C.; LARSON, J.E.; DiCOSTANZO, A. Effects of feeding polyclonal antibody preparations on rumen fermentation patterns, performance, and carcass characteristics of feedlot steers. Journal of Animal Science, v.86, n.11, p.3023-3032, 2008.
  • EDWARDS, J.E.; MCEWAN, N.R.; TRAVIS, A.J.; WALLACE, R.J. 16S rDNA library-based analysis of ruminal bacterial diversity. Antonie Van Leeuwenhoek, v.86, p.263-281, 2004.
  • FOX, D.G.; TEDESCHI, L.O.; TYLUTKI, T.P.; RUSSELL, J.B.; Van AMBURGH, M.E.; CHASE, L.E.; PELL, A.N.; OVERTON, T.R. The Cornell Net Carbohydrate and Protein System model for evaluating herd nutrition and nutrient excretion. Animal Feed Science Technology, v.112, n.1-4, p.29-78, 2004.
  • FRANZOLIN, R.; DEHORITY, B.A. Effect of prolonged high concentrate feeding on ruminal protozoa concentrations. Journal of Animal Science, v.74, n.11, p.2803-2809, 1998.
  • GRIFFITHS, R.I.; WHITELEY, A.S.; O'DONNELL, A.G.O.; BAILEY, M.J. Rapid method for coextraction of DNA and RNA from natural environments for analysis of ribosomal DNA- and RNA-based microbial community composition. Applied Environmental Microbiology, v.66, n.12, p.5488-5491, 2000.
  • HALL, M.B. Recent advanced in nonndf carbohydrates for the nutrition of lactating cows. In: INTERNATIONAL SYMPOSIUM IN DAIRY CATTLE, 2001, Lavras, MG. Anais.. Lavras, MG: UFLA/FAEPE, 2001. p.139-148.
  • HARDY, B. The issue of antibiotic use in livestock industry. What have we learned? Animal Biotechnology, v.13, n.1, p.129-147, 2002.
  • HEUER, H.: KRSEK, M.; BAKER, P.; SMALLA, K.; WELLINGTON, E.M.H. Analysis of Actinomycete communities by specific amplification of genes encoding 16S rRNA and gel-electrophoretic separation in denaturing gradients. Applied and Environmental Microbiology, v.63, n.8, p.3233-3241, 1997.
  • JANSSEN, P.H.; KIRS, M. Structure of the Archaeal community of the rumen. Applied and Environmental Microbiology, v.74, n.12, p.3619-3625, 2008.
  • KOCHERGINSKAYA, S.A.; AMINOV, R.I.; WHITE, B.A. Analysis of the rumen bacterial diversity under two different diet conditions using denaturing gradient gel electrophoresis, random sequencing, and statistical ecology approaches. Anaerobe, v.7, n.3, p.119-134, 2001.
  • KONONOFF, P.J.; HEINRICHS, A.J.; BUCKMASTER, D.R. Modification of the Penn State Forage and Total Mixed Ration Particle Separator and the effects of moisture content on its measurements. Journal of Dairy Science, v.86, n.5, p.1858-1863, 2003.
  • KUDO, Y.; NAKAJIMA, T.; MIYAKI, T.; OYAIZU, H. Methanogen flora of paddy soils in Japan. FEMS Microbiology Ecology, v.22, n.1, p.39-48, 1997.
  • MARINO, C.T.; OTERO, W.G.; RODRIGUES, P.H.M.; DICOSTANZO, A.; MILLEN, D.D.; PACHECO, R.D.L.; DILORENZO, N.; MARTINS, C.L.; ARRIGONI, M.D.B. Effects of adding polyclonal antibody preparation on ruminal fermentation patterns and digestibility of cows fed different energy sources. Journal of Animal Science, v.89, n.5, p.3228-3235, 2011.
  • MARTINELE, I.; SIQUEIRACASTRO, I.C.V.; D'AGOSTO, M. Protozoários ciliados no rúmen de bovinos alimentados com dietas de capim-elefante e dois níveis de concentrado. Revista Brasileira de Sáude e Produção Animal [Online], v.9, n.1, p.74-81, 2008.
  • MILLEN, D.D. Performance, rumen papillae evaluation and metabolic profile of feedlot cattle supplemented with sodic monensin or polyclonal antibodies 2008. 131p. MSc Diss. São Paulo State University, São Paulo.
  • MIN, B.R.; PINCHAK, W.E.; ANDERSON, R.C.; HUME, M.E. In vitro bacterial growth and in vivo ruminal microbiota populations associated with bloat in steers grazing wheat forage. Journal of Animal Science, v.84, n.10, p.2873-2882, 2006.
  • MUYER, G. DGGE/TGGE, a method for identifying genes from natural communities. Current Opinion in Microbiology, v.2, n.3, p.317-322, 1999.
  • NÜBEL, U.; ENGELEN, B.; FELSKE, A.; SNAIDR, J.; WIESHUBER, A.; AMANN, R.I.; LUDWIG, W.; BACKHAUS, H.J. Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis. Journal of Bacteriology, v.178, n.19, p.5636-5643, 1996.
  • SAS INSTITUTE. User's guide: statistics Version 9.2. Cary, 2008.
  • SILVA, D.J.; QUEIROZ, A.C. Análise de alimentos: métodos químicos e biológicos 2ed. Viçosa: Universidade Federal de Viçosa, 2002. 165p.
  • TAJIMA, K.; AMINOV, R.I.; NAGAMINE, T.; MATSUI, H.; NAKAMURA, M.; BENNO, Y. Dietdependent shifts in the bacterial population of the rumen revealed with real-time PCR. Applied Environmental Microbiology, v.67, n.6, p.2766-2774, 2001.
  • VAN SOEST, P.J.; ROBERTSON, J.B.; LEWIS, B.A. Symposium: Carbohydrate methodology, metabolism, and nutritional implications in dairy cattle. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. Journal of Dairy Science, v.74, n.10, p.3583-3597, 1991.
  • WILLIAMS, A.G.; COLEMAN, G.S. The rumen protozoa. In: HOBSON, P.N. The rumen microbial ecosystem London, UK: Elsevier Applied Science, 1988. p.77-128.

Publication Dates

  • Publication in this collection
    29 Sept 2014
  • Date of issue
    June 2012

History

  • Received
    11 Apr 2011
  • Accepted
    19 Mar 2012
UFBA - Universidade Federal da Bahia Avenida Adhemar de Barros nº 500 - Ondina , CEP 41170-110 Salvador-BA Brasil, Tel. 55 71 32836725, Fax. 55 71 32836718 - Salvador - BA - Brazil
E-mail: rbspa@ufba.br