[Truncated abstract] Antimicrobial growth promoters are added to feed to prevent lactic acidosis in ruminant animals by selectively inhibiting rumen bacteria that produce lactic acid. However, recently imposed or impending bans on the use of antimicrobial growth promoters in animal production have lead to a critical need to find practical alternatives that are safe for the animal and consumer and that obtain similar production benefits. I investigated bioactive plants of Australia for their potential to prevent lactic acidosis in ruminants. The unifying hypothesis tested was that plants would be identified that selectively inhibit lactic acid-producing bacteria and consequently protect against lactic acidosis. This hypothesis was tested in a three phase process: phase 1, plant selection and collection; phase 2, a three stage protocol for screening plants and essential oils; phase 3, in vivo experiments and chemical fractionation of the most promising plant. I developed an in vitro bioassay that simulated acidosis by adding glucose to rumen fluid in Bellco tubes and incubating for 5 h (Chapter 4). The pH and gas production were used as indicators of acidosis and fermentation activity. I used this bioassay to screen ninety-five plants (dried and ground material from 79 species) and ten essential oils and included a negative control (oaten chaff) and a positive control (virginiamycin). One plant, Eremophila glabra, produced a similar pH (5.63) to the positive control (5.43) although it inhibited gas production to a moderate extent (P < 0.05). ... Seven serrulatane diterpenes were identified to be the major secondary metabolites in E. glabra. The metabolites were screened using a broth dilution and microtitre spectrophotometry method and were selective against S. bovis at between 320 and 1077 [mu]g/ mL. The serrulatanes from E. glabra were probably responsible for the activity against acidosis that I observed in vitro, because they selectively inhibited lactateproducing bacteria. It is also possible that a synergy between serrulatanes and possibly other metabolites are responsible for the activity observed in vitro. The results from my experiments support the role that bioactive plants may have to replace the antibiotics that are added to livestock feed. Australian plants were identified containing compounds that were active against the bacterial processes responsible for ruminant acidosis. To my knowledge this is the first work undertaken to identify bioactive plants of Australia for their potential to prevent acidosis. I developed in vitro screening bioassays that targeted key indicators of acidosis. These bioassays enabled me to identify 5 plants from the 104 screened that could potentially control acidosis. One of these plants in particular, E. glabra, showed a level of activity in vitro that was comparable to antibiotic protection against acidosis. The exciting in vitro results were not demonstrated in vivo but only one dose level of E. glabra was used, which was based on the in vitro work. In contrast to the in vitro system the rumen is a continuous flow system with greater complexity and it is possible that the concentration of E. glabra that I used in vivo was not optimum. This places importance on future dose response experiments to confirm the efficacy of E. glabra in vivo.

Made available in DSpace on 2015-03-26T13:55:24Z (GMT). No. of bitstreams: 1 texto completo.pdf: 614632 bytes, checksum: eac726a0efd1eec87578c9745a474e81 (MD5) Previous issue date: 2014-02-26 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / This work aimed to evaluate the effects of ruminal and/or abomasal protein supplementation in cattle fed tropical forages on nutritional variables, degradation of insoluble fiber, activity of fibrolytic enzymes and rumen bacterial communities profile. Two experiments were carried out sequentially, and in the same way, differing only on the forage quality that was defined based on crude protein content: medium quality (784 g of crude protein/ kg of dry matter) and high quality (986 g of crude protein/ kg of dry matter). Four Nellore young bulls, averaging 227±11 kg body weight and fitted with ruminal and abomasal cannullas, were used. The following treatments were evaluated: control (only forage), ruminal nitrogenous compounds supplementation (daily supply of 230 g of crude protein in the rumen), abomasal nitrogenous compounds supplementation (daily supply of 230 g of crude protein in the abomasum), and ruminal and abomasal nitrogenous compounds supplementation (daily supply of 230 g of protein, being 115 g in the rumen and 115 g in the abomasum). Casein was used as protein source for supplementation and basal diet consisted of Tifton 85 (Cynodon spp) hay. Two 4 x 4 Latin squares experimental trials, balanced for residual effects, with four treatments, four animals and four experimental periods lasting 29 days each, were implemented sequentially, one for each forage as stated above. There were no effects (P>0.05) of supplementation scheme or forage quality on organic matter (OM) intake. On the other hand, neutral detergent fiber (NDF) intake was higher (P<0.05) when animals were fed high quality forage. The total digestibility of OM was increased (P<0.05) by supplementation and by increasing forage quality. The digestibility coefficient of NDF was not affected by supplementation scheme (P>0.05), but was increased by the enhancing in forage quality. The apparent ruminal nitrogen balance (RNB) was increased (P<0.05) when animals were supplemented in rumen or in both rumen and abomasum cannula. It is worth highlighting that no supplementation or exclusively abomasal supplementation led to negative values of RNB. The apparent nitrogen balance (NB) was increased (P>0.05) when high quality forage was fed to the animals and by supplementation, no matter if the supplement was infused into rumen, abomasum or both. Ruminal supplementation enhanced (P<0.05) the concentration of rumen ammonia nitrogen (RAN). Although, animals supplemented in rumen and abomasum or exclusively in abomasum did not exhibit increase in RAN concentrations (P>0.05) when compared to no supplemented animals. The bacterial richness and diversity (Shannon-Wiener index) associated with the solid phase of rumen digesta were not affected (P>0.05). However, richness and diversity associated with the liquid phase were increased (P<0.05) by the improvement in forage quality. NDF degradation rate (kp) was enhanced (P<0.05) by ruminal supplementation. Carboximetilcellulase activity was decreased (P<0.05) by the abomasal and ruminal/abomasal supplementation. Xylanase activity was decreased (P<0.05) by both post-ruminal supplementation schemes, when high quality forage was fed to the animals. It was not detected correlations among variables associated with bacterial diversity and activity and variables associated with nutritional performance, excepted for the positive correlation between kp and RNB Clustering analyses based on the Unweighted Pair Group Method (UPGMA) evidenced that distinct bacterial communities are associated with liquid and solid fraction, independently of the forage quality. Clustering analyses also showed that forage quality affects the diversity of liquid phase associated bacteria, even though, effects on solid phase was not clear. None of the evaluations related to the microbial community were able to detect clear effects of supplementation on the microbial community diversity. Improving forage quality enhances fiber digestibility, nitrogen balance and modifies the bacterial community associated with liquid phase. On the other hand, protein supplementation enhances the nitrogen retention in animal body. / Objetivou-se avaliar os efeitos da suplementação proteica ruminal e/ou abomasal em bovinos consumindo forragens tropicais sobre as características nutricionais, a atividade enzimática fibrolítica, a degradação in vitro da fibra insolúvel e o perfil da comunidade bacteriana ruminal. Dois experimentos foram realizados sequencialmente diferindo apenas na qualidade do feno ofertado, sendo de média qualidade (78,4 g de proteína bruta/kg de matéria seca) e alta qualidade (98,6 g de proteína bruta/kg de matéria seca), respectivamente. Os procedimentos experimentais foram os mesmos para ambas as forragens. Foram utilizados quatro novilhos Nelore, não castrados, com peso corporal médio de 227±11 kg, fistulados no rúmen e no abomaso. Foram avaliados os seguintes esquemas de suplementação: controle (somente forragem); suplementação nitrogenada ruminal (fornecimento diário de 230 g de proteína suplementar no rúmen); suplementação nitrogenada abomasal (fornecimento diário de 230 g de proteína suplementar no abomaso) e suplementação nitrogenada ruminal e abomasal (fornecimento diário de 230 g de proteína suplementar, sendo 115 g no rúmen e 115 g no abomaso). A alimentação volumosa basal foi constituída por feno de tifton 85 (Cynodon spp) e como fonte de compostos nitrogenados suplementares foi utilizada a caseína (caseína pura, Labsynth). O experimento foi conduzido segundo delineamento em quadrado latino 4 x 4 balanceado para efeitos residuais, sendo dois quadrados sequenciais, um para cada forragem, com quatro esquemas de suplementação, quatro animais e quatro períodos experimentais com 29 dias em cada quadrado. Não foram observados efeitos (P>0,05) da qualidade da foragem ou do esquema de suplementação sobre o consumo de matéria orgânica (MO). Por outro lado, o consumo de fibra em detergente neutro (FDN) foi superior (P<0,05) quando forragem de alta qualidade foi ofertada aos animais. O coeficiente de digestibilidade total da MO foi incrementado (P>0,05) pela suplementação e pela melhoria na qualidade da forragem. O coeficiente de digestibilidade da FDN não foi afetado pelo esquema de suplementação (P>0,05), mas foi superior quando os animais receberam forragem de alta qualidade (P<0,05). O balanço aparente de compostos nitrogenados no rúmen (BNR) foi ampliado (P<0,05) pela suplementação no rúmen e no rúmen-abomaso. Ressalta-se que a ausência de suplementação ou a suplementação exclusiva no abomaso proporcionam valores negativos de BNR. O balaço aparente de compostos nitrogenados no organismo animal foi ampliado (P<0,05) pelo fornecimento de suplemento, independentemente do local de suplementação, e pelo fornecimento de forragem de melhor qualidade A suplementação no rúmen incrementou (P<0,05) a concentração de nitrogênio amoniacal ruminal (NAR). Por outro lado, animais recebendo suplementação no rúmen e abomaso ou exclusivamente no abomaso apresentaram concentrações de NAR similares (P>0,05) a animais não suplementados. A riqueza e a diversidade microbiana da fase sólida da digesta ruminal não foram afetadas (P>0,05). Entretanto, a riqueza e a diversidade bacteriana associada à fase líquida da digesta ruminal foram incrementadas (P<0,05) pela melhoria na qualidade da forragem. A taxa de degradação da FDN foi ampliada (P<0,05) pela suplementação no rúmen. A atividade da carboximetilcelulase foi reduzida (P<0,05) pela suplementação no abomaso e rúmen-abomaso. A atividade da xilanase foi deprimida (P<0,05) pela suplementação pós-ruminal somente quando forragem de alta qualidade foi fornecida aos animais. Não foi observada correlação (P>0,05) entre as variáveis associadas à atividade enzimática e diversidade microbiana e as características ligadas ao desempenho nutricional, exceção feita à correlação positiva (P<0,05) entre a taxa de degradação da FDN e o BNR. A avaliação multivariada da diversidade permitiu evidenciar que comunidades bacterianas distintas estão associadas às frações líquida e sólida, independente da qualidade da forragem ofertada. Percebeu- se que a variação na qualidade da forragem influencia a diversidade microbiana associada à fração líquida do fluido ruminal, embora não tenha exercido tal efeito sobre a fração sólida. Em nenhuma avaliação percebeu-se efeito claro do esquema de suplementação sobre o perfil da comunidade microbiana ruminal. A melhoria da qualidade da forragem amplia a digestibilidade da fibrae a retenção de nitrogênio e é capaz de modificar a comunidade bacteriana associada à fração liquida do conteúdo ruminal. A suplementação proteica, por sua vez, independentemente do local de suplementação, amplia a retenção de nitrogênio no organismo animal.