In vitro evaluation, in vivo quantification, and microbial diversity studies of nutritional strategies for reducing enteric methane production

The main objective of the present work was to study nutritive strategies for lessening the CH4 formation associated to ruminant tropical diets. In vitro gas production technique was used for evaluating the effect of tannin-rich plants, essential oils, and biodiesel co-products on CH4 formation in three individual studies and a small chamber system to measure CH4 released by sheep for in vivo studies was developed. Microbial rumen population diversity from in vitro assays was studied using qPCR. In vitro studies with tanniniferous plants, herbal plant essential oils derived from thyme, fennel, ginger, black seed, and Eucalyptus oil (EuO) added to the basal diet and cakes of oleaginous plants (cotton, palm, castor plant, turnip, and lupine), which were included in the basal diet to replace soybean meal, presented significant differences regarding fermentation gas production and CH4 formation. In vivo assays were performed according to the results of the in vitro assays. , when supplemented to a basal diet (Tifton-85 hay sp, corn grain, soybean meal, cotton seed meal, and mineral mixture) fed to adult Santa Ines sheep reduced enteric CH4 emission but the supplementation of the basal diet with EuO did not affect ( > 0.05) methane released. Regarding the microbial studies of rumen population diversity using qPCR with DNA samples collected from the in vitro trials, the results showed shifts in microbial communities of the tannin-rich plants in relation to control plant. This research demonstrated that tannin-rich , essential oil from eucalyptus, and biodiesel co-products either in vitro or in vivo assays showed potential to mitigate CH4 emission in ruminants. The microbial community study suggested that the reduction in CH4 production may be attributed to a decrease in fermentable substrate rather than to a direct effect on methanogenesis.

Comparative in vitro evaluation of forage legumes (prosopis, acacia, atriplex, and leucaena) on ruminal fermentation and methanogenesis

Two experiments in vitro were conducted to evaluate four Egyptian forage legume browses, i.e., leaves of prosopis (Prosopis juliflora), acacia (Acacia saligna), atriplex (A triplex halimus), and leucaena (Leucaena leucocephala), in comparison with Tifton (Cynodon sp.) grass hay for their gas production, methanogenic potential, and ruminal fermentation using a semi-automatic system for gas production (first experiment) and for ruminal and post ruminal protein degradability (second experiment). Acacia and leucaena showed pronounced methane inhibition compared with Tifton, while prosopis and leucaena decreased the acetate:propionate ratio (P<0.01). Acacia and leucaena presented a lower (P<0.01) ruminal NH3-N concentration associated with the decreasing (P<0.01) ruminal protein degradability. Leucaena, however, showed higher (P<0.01) intestinal protein digestibility than acacia. This study suggests that the potential methanogenic properties of leguminous browses may be related not only to tannin content, but also to other factors.

Effects of Aspergillus spp. exogenous fibrolytic enzymes on in vitro fermentation of tropical forages

BACKGROUND: Cellulose and hemicellulose are quantitatively the most important structural carbohydrates present in ruminant diets. Rumen micro-organisms produce enzymes that catalyse their hydrolysis, but the complex network formed by structural carbohydrates and lignin reduces their digestibility and restricts efficient utilisation of feeds by ruminants. This study aimed to produce two enzymatic extracts, apply them in ruminant diets to determine the best levels for ruminal digestibility and evaluate their effects on in vitro digestibility. RESULTS: In experiment 1 a two-stage in vitro technique was used to examine the effects of different enzymatic levels of Aspergillus japonicus and Aspergillus terricola on tropical forages. Enzyme addition had minor effects on corn silage at the highest enzymatic level. In experiment 2 an in vitro gas production (GP) technique was applied to determine apparent in vitro organic matter digestibility and metabolisable energy. The addition of enzymes in GP showed interesting results. Good data were obtained using sugar cane and Tifton-85 hay supplemented with extracts of A. japonicus and A. terricola respectively. CONCLUSION: Overall, the study suggests that addition of crude extracts containing exogenous fibrolytic enzymes to ruminant diets enhances the effective utilisation of ruminant feedstuffs such as forages. Copyright (c) 2012 Society of Chemical Industry

Streptomyces herbaceus sp nov., Streptomyces incanus sp nov and Streptomyces pratens sp nov., isolated from the soil of a hay meadow

The taxonomic positions of three streptomycetes isolated from a soil sample from a hay meadow were determined using a polyphasic approach. The isolates had chemical and morphological properties typical of the genus Streptomyces and, in phylogenetic analyses based on 16S rRNA gene sequences, formed a distinct subclade that was most closely related to the Streptomyces prasinus subclade. DNA-DNA relatedness studies showed that the novel strains belonged to three different genomic species. The novel strains could be distinguished from one another and from the type strains of the species classified in the S. prasinus subclade using a combination of genotypic and phenotypic properties. On the basis of these data, it is proposed that the novel strains be assigned to the genus Streptomyces as Streptomyces herbaceus sp. nov., Streptomyces incanus sp. nov. and Streptomyces pratens sp. nov., with BK119(T) (=KACC 21001(T) =CGMCC 4.5797(T)), BK128(T) (=KACC 21002(T) =CGMCC 4.5799(T)) and BK138(T) (=KACC 20904(T) =CGMCC 4.5800(T)) as the respective type strains.

Comparison of indigestible markers from in situ and in vivo incubation to predict apparent digestibility in hay- and corn-fed horses

Four castrated crossbred horses were used in a randomized block design to study the use of indigestible internal markers iNDF and iADF obtained in situ (from bovines) or in vivo (from equines). Treatments consisted of determining digestibility by the direct method comprising total feces collection (TC) and by the indirect method comprising internal markers iNDF and iADF obtained by in situ incubation in bovine rumen or in vivo by the mobile nylon bag (MNB) technique with horses. iNDF-IV and iADF-IV resulted in better marker recovery rate (RR) (91.50%), similar to TC. The in situ technique resulted in lower RR values for the two indigestible markers, averaging 86.50% (p < 0.05). Estimates of the nutrient coefficient of digestibility (CD) were adequately predicted by iADF-IV, for horses fed on hay exclusively, with rates 46.41, 48.16, 47.92 and 45.51% for dry matter (DM), organic matter (OM), FDN and gross energy, respectively. Results show that MNB may be used to obtain iADF in horses fed on coast-cross hay exclusively, whereas NDFi and ADFi were selected for horses fed on mixed diets to predict the coefficient of nutrient digestibility.