A lack of predatory interaction between rumen ciliate protozoa and Shiga-toxin producing Escherichia coli

Aims: To investigate interactions between rumen protozoa and Shiga toxin-producing Escherichia coli (STEC) and to ascertain whether it is likely that rumen protozoa act as ruminant hosts for STEC. Methods and Results: The presence of stx genes in different microbial fractions recovered from cattle and sheep rumen contents and faeces was examined using PCR. In animals shedding faecal STEC, stx genes were not detected in the rumen bacterial or rumen protozoal fractions. Direct interactions between ruminal protozoa and STEC were investigated by in vitro co-incubation. Rumen protozoa did not appear to ingest STEC, a STEC lysogen or non-STEC E. coli populations when co-incubated. Conclusions: The ruminal environment is unlikely to be a preferred habitat for STEC. Bacterial grazing by rumen protozoa appears to have little, if any, effect on STEC populations. Significance and Impact of the Study: This study indicates that ruminal protozoa are unlikely to be a major factor in the survival of STEC in ruminants. They appear as neither a host that protects STEC from the ruminal environment nor a predator that might reduce STEC numbers.

Organophosphonates revealed: new insights into the microbial metabolism of ancient molecules.

Organophosphonates are ancient molecules that contain the chemically stable C–P bond, which is considered a relic of the reducing atmosphere on primitive earth. Synthetic phosphonates now have a wide range of applications in the agricultural, chemical and pharmaceutical industries. However, the existence of C–P compounds as contemporary biogenic molecules was not discovered until 1959, with the identification of 2-aminoethylphosphonic acid in rumen protozoa. Here, we review advances in our understanding of the biochemistry and genetics of microbial phosphonate metabolism, and discuss the role of these compounds and of the organisms engaged in their turnover within the P cycle.

Efeitos de rações com níveis crescentes de cana-de-açúcar em substituição à silagem de milho sobre a população de protozoários ciliados no rúmen de ovinos

Oito ovinos machos com fístulas ruminais foram delineados em dois quadrados latinos, em quatro períodos de 35 dias cada e quatro tratamentos, compreendendo níveis crescentes de cana-de-açúcar (CA) em substituição à silagem de milho (SM): A) 100% SM; B) 67% SM e 33% CA; C) 33% SM e 67% CA; e D 100% CA, para avaliação da população de protozoários ciliados no rúmen. As concentrações totais de protozoários ciliados no rúmen e de Entodinium diminuíram e os valores de pH do conteúdo ruminal aumentaram linearmente com o incremento das quantidades de cana-de-açúcar na ração. Não houve diferenças entre tratamentos para o volume ruminal e a taxa de passagem da fase líquida do rúmen.

População protozoários ciliados e degradabilidade ruminal em búfalos e bovinos zebuínos sob dieta à base de cana-de-açúcar

Três búfalos e três bovinos zebuínos adultos com cânulas ruminais foram alimentados à vontade com dieta de cana-de-açúcar fresca e picada, suplementada com 3 kg de concentrado/animal, durante 13 semanas. Após duas semanas de adaptação, amostras do conteúdo ruminal foram coletadas semanalmente, em dois tempos de amostragem: antes da alimentação e 40 minutos após para determinação da concentração e composição da fauna ruminal. Na quinta semana do experimento, os conteúdos ruminais foram misturados e reinoculados entre animais de mesma espécie e, na última semana, estimadas as degradabilidades in situ da MS, PB e FDN da cana-de-açúcar e MS e PB do concentrado. A concentração média de protozoários foi maior em bovinos, de 4,85 x 10(5)/mL, que em búfalos, de 3,82 x 10(5)/mL. As composições genéricas para Entodinium e subfamília Diplodiniinae foram 79,2 e 6,2% em bovinos e 32,0 e 54,9% em búfalos, respectivamente. Não houve interação significativa entre espécies animais e tempo de amostragem. Houve aumento da concentração média dos holotricos após 40 minutos da alimentação. As degradabilidades efetivas dos nutrientes estudados foram semelhantes entre as duas espécies animais, observando-se diferenças na cinética da degradação. Concluiu-se que os búfalos apresentaram menor concentração de ciliados no rúmen, maior composição de Diplodiniinae e menor de Entodinium que os bovinos; os holotricos exibiram capacidade de migração e seqüestro no rúmen em ambas as espécies; e as diferenças na fauna ruminal não influenciaram a degradabilidade efetiva dos nutrientes da cana-de-açúcar e do concentrado.

Efeitos de Dietas com Diferentes Níveis de Fibra em Detergente Neutro e do Tamanho de Poros de Sacos de Náilon Incubados no Rúmen sobre a Fauna Ruminal em Bubalinos e Bovinos

Quatro bubalinos e quatro bovinos, fistulados no rúmen, foram distribuídos aleatoriamente a dois quadrados latinos (4 x 4) para avaliar os efeitos de rações com quatro níveis de FDN (54, 60, 66 e 72%) e de dois tamanhos de poros em sacos de náilon (53 e 100 mm) sobre a população de protozoários ciliados no conteúdo ruminal e dentro dos sacos incubados em diversos tempos. Não houve diferença nos diversos gêneros estudados e no total de protozoários nas amostras dos sacos de náilon entre 53 e 100 mm. As concentrações de Entodinium e de total de protozoários do conteúdo ruminal elevaram-se até o nível de 66% de FDN, com queda acentuada em 72% nos bubalinos, enquanto nos bovinos houve redução com o aumento de FDN na ração. Houve diferenças nas concentrações dos protozoários da subfamília Diplodiniinae, Epidinium e Dasytricha entre bubalinos e bovinos, dentro das amostras incubadas em sacos com poros de 53 mm. A concentração de Diplodiniinae aumentou com 72% de FDN na dieta em ambas as espécies animais. Os bubalinos apresentaram valores médios de pH e taxa de passagem do líquido ruminal mais elevados e volumes ruminais menores que os bovinos.

Efeitos de dietas com polpa cítrica em substituição ao milho em grãos no concentrado sobre a degradabilidade e a fauna ruminal em bubalinos

Quatro búfalos com cânulas ruminais foram distribuídos aleatoriamente em quadrado latino 4 x 4, compreendendo quatro rações com feno de capim-coastcross (Cynodon dactylon) (50% MS) e níveis crescentes de polpa cítrica peletizada em substituição ao milho em grão moído no concentrado (0, 33, 66 e 100% MS). Foram avaliadas a degradabilidade dos nutrientes e a população de protozoários no rúmen. Houve efeito quadrático da fração solúvel dos nutrientes entre os tratamentos, com valores maiores nos dois extremos. A taxa de degradação da PB do feno e da MS da polpa cítrica apresentou-se linear com aumento da polpa cítrica na ração. As rações contendo polpa cítrica promoveram menores concentrações de Entodinium e número total de ciliados/mL que a dieta só com milho. Houve predominância dos Entodinium nas dietas contendo somente milho ou somente polpa cítrica, enquanto em mistura predominaram os ciliados da subfamília Diplodiniinae. Observou-se efeito de interação da polpa cítrica com o milho, promovendo modificações no metabolismo ruminal, especialmente na solubilidade dos nutrientes, na curva do pH e na composição da fauna.

Effects of different lipid levels on protozoa population, microbial protein synthesis and rumen degradability in cattle

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A dynamic mechanistic model of lactic acid metabolism in the rumen

Current feed evaluation systems for ruminants are too imprecise to describe diets in terms of their acidosis risk. The dynamic mechanistic model described herein arises from the integration of a lactic acid (La) metabolism module into an extant model of whole-rumen function. The model was evaluated using published data from cows and sheep fed a range of diets or infused with various doses of La. The model performed well in simulating peak rumen La concentrations (coefficient of determination = 0.96; root mean square prediction error = 16.96% of observed mean), although frequency of sampling for the published data prevented a comprehensive comparison of prediction of time to peak La accumulation. The model showed a tendency for increased La accumulation following feeding of diets rich in nonstructural carbohydrates, although less-soluble starch sources such as corn tended to limit rumen La concentration. Simulated La absorption from the rumen remained low throughout the feeding cycle. The competition between bacteria and protozoa for rumen La suggests a variable contribution of protozoa to total La utilization. However, the model was unable to simulate the effects of defaunation on rumen La metabolism, indicating a need for a more detailed description of protozoal metabolism. The model could form the basis of a feed evaluation system with regard to rumen La metabolism.

Effects of dry sugar cane yeast on the diet intake, digestibility and bovine rumen microbial populations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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

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.

Digestibility, fermentation and rumen microbiota of crossbred heifers fed diets with different soybean oil availabilities in the rumen

The goal of this study was to evaluate the effects of different soybean oil availabilities on the intake and partial and total digestibility of dry matter (DM) and nutrients, rumen fermentation parameters, efficiency of microbial synthesis, and the rumen microbiota of crossbred beef heifers. Nine crossbred heifers fitted with rumen and duodenal cannulae were evaluated in a triple 3 × 3 Latin square design with three treatments and three periods in three simultaneous repetitions. Heifers approximately 18 months old, with mean initial and final body weights of 316.3±28.8 and 362.6±34.4 kg, respectively, were fed a diet containing 600. g/kg of corn silage and 400. g/kg concentrate with a 58.0. g/kg fat content in the total diet. The sources of lipids included soybean grain, rumen-protected fat, and soybean oil. The statistical analyses were conducted using PROC MIXED from SAS, and the means were compared using Tukey's test (P<0.05). Dietary lipid sources did not affect nutrient intake (P>0.05). Except the apparent digestibility of organic matter (P=0.024), the apparent digestibility of the other nutrients did not differ among the treatment groups. Regarding body nitrogen retention, the soybean grain treatment was more effective than the rumen-protected fat treatment (P=0.045); however, the soybean oil treatment did not differ from the other two treatments. In relation to the efficiency of microbial protein synthesis (g. N/kg of organic matter apparently digested in the rumen corrected for microbial organic matter), the soybean oil and soybean grain treatments were more efficient than the rumen-protected fat treatment (P=0.001). Animals fed rumen-protected fat had larger numbers of protozoa (P<0.001) and fungi (P<0.001) than those supplemented with soybean grain and soybean oil. The dietary lipid sources did not affect pH, the molar concentration of propionate and total volatile fatty acids (P>0.05), whereas the concentrations of ammonia nitrogen and acetate were higher in animals fed with rumen-protected fat than in those submitted to the other treatments. The use of different soybean oil availabilities did not affect nutrient intake; however, treatments with soybean oil and soybean grain were more efficient regarding nutrient intake than rumen-protected fat because they reduced the numbers of fungi and protozoa and consequently improved the efficiency of microbial protein synthesis. © 2013 Elsevier B.V.

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

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Feeding increasing concentrate to Tifton 85 hay ratios modulated rumen fermentation and microbiota in Nellore feedlot steers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cellulosic waste degradation by rumen-enhanced anaerobic digestion

Anaerobic digestion of lignocellulosic material is carried out effectively in many natural microbial ecosystems including the rumen. A rumen-enhanced anaerobic sequencing batch reactor was used to investigate cellulose degradation to give analysis of overall process stoichiometry and rates of hydrolysis. The reactor achieved VFA production rates of 207-236 mg COD/L/h at a loading rate of 10 g/L/d. Overloading of the reactor resulted in elevated production of propionic acid, and on occasion, the presence of succinic acid. With improvements in mixing and solids wasting, the anaerobic sequencing batch reactor system could enable full-scale application of the process for treatment of cellulosic waste material.

Ecology of uncultivated oscillospira species in the rumen of cattle, sheep, and reindeer as assessed by microscopy and molecular approaches

The ecology of the uncultured, but large and morphologically conspicuous, rumen bacterium Oscillospira spp. was studied. Oscillospira-specific 16S rRNA gene sequences were detected in North American domestic cattle, sheep from Australia and Japan, and Norwegian reindeer. Phylogenetic analysis of the sequences obtained allowed definition of three operational taxonomic units within the Oscillospira clade. Consistent with this genetic diversity, we observed atypical smaller morphotypes by using an Oscillospira-specific fluorescence in situ hybridization probe. Despite the visual disappearance of typical large Oscillospira morphotypes, the presence of Oscillospira spp. was still detected by Oscillospira-specific PCR in the rumen of cattle and sheep. These observations suggest the broad presence of Oscillospira species in various rumen ecosystems with the level, and most likely the morphological form, dependent on diet. An ecological analysis based on enumeration of the morphologically conspicuous, large-septate form confirms that the highest counts are associated with the feeding of fresh forage diets to cattle and sheep and in two different subspecies of reindeer investigated.