Interaction between methanotrophy and gastrointestinal nematodes infection on the rumen microbiome of lambs.

Complex cross-talk occurs between gastrointestinal nematodes and gut symbiotic microbiota, with consequences for animal metabolism. To investigate the connection between methane production and endoparasites, this study evaluated the effect of mixed infection with Haemonchus contortus and Trichostrongylus colubriformis on methanogenic and methanotrophic community in rumen microbiota of lambs using shotgun metagenomic and real-time quantitative PCR (qPCR). The rumen content was collected from six Santa Inês lambs, (7 months old) before and after 42 days infection by esophageal tube. The metagenomic analysis showed that the infection affected the microbial community structure leading to decreased abundance of methanotrophs bacteria, i.e. α-proteobacteria and ß-proteobacteria, anaerobic methanotrophic archaea (ANME), protozoa, sulfate-reducing bacteria, syntrophic bacteria with methanogens, geobacter, and genes related to pyruvate, fatty acid, nitrogen, and sulfur metabolisms, ribulose monophosphate cycle, and Entner-Doudoroff Pathway. Additionally, the abundance of methanogenic archaea and the mcrA gene did not change. The co-occurrence networks enabled us to identify the interactions between each taxon in microbial communities and to determine the reshaping of rumen microbiome associations by gastrointestinal nematode infection. Besides, the correlation between ANMEs was lower in the animal's postinfection. Our findings suggest that gastrointestinal parasites potentially lead to decreased methanotrophic metabolism-related microorganisms and genes.

Potential use of cassava by-product as ruminant feed.

Cassava (Manihot esculenta Crantz) bagasse is the by-product from industry (BCI), generated during manufacturing of cassava flour; this material has significant amounts of carbohydrates consisting in a potential energy source for ruminants. We hypothesized that the inclusion of BCI in the diets may lead to fermentation parameters equivalent to those of conventional feedstuff such as tropical grasses or grains; therefore, we aimed to evaluate ruminal fermentation parameters of BCI in in vitro conditions. Three different substrates were prepared: 100% BCI (BCI diet), 100% tifton (Cynodon spp.) hay (CTL diet), and 50% tifton hay +50% BCI (THB diet). Ruminal fermentation parameters of these diets were evaluated in in vitro gas production assays. In a 24-h incubation, increased values for total gas production, organic matter degradability, and methane production were observed for BCId and THB as compared to CTL (p < 0.05), while neutral THB showed the highest value for neutral detergent fiber degradability (p < 0.05). Fermentation profile was evaluated in a 48-h assay: shorter lag time as well as increased gas production potential and fractional fermentation rate were observed for the BCId and THB as compared to CTL (p < 0.05). Our results suggested that by-product from cassava industry is a suitable feed for ruminant production, providing desirable in vitro ruminal fermentation performance and organic matter degradability.

Fibrolytic enzymes improving in vitro rumen degradability of tropical forages.

During specific times of the year, especially dry seasons, tropical forages typically have poor nutritional value due to high contents of neutral and acid detergent fibres, and low crude protein in their composition, which may reduce productivity of ruminant livestock production and lead to increased enteric methane (CH4 ) emissions per unit of generated product in forage-bases systems. In order to increase fibre degradability and the efficiency of energy utilisation from low-quality forages, exogenous fibrolytic enzymes have been studied. In this assay, we evaluated the effects of increasing dose levels of fibrolytic enzymes extract (FEE) produced by Trichoderma reesei on in vitro rumen organic matter degradability, fermentation parameters, total gas and CH4 production of tropical forages. Forage samples were analysed for their bromatological composition, and enzyme activity from FEE was performed for xylanase and endoglucanase. The in vitro gas production technique was used in a 5 × 3 factorial arrangement with five FEE dose levels (0, 5, 50, 500 and 5,000 µl) and three substrates (Cynodon spp., Panicum maximum and Cenchrus ciliaris L.). The highest dose level of FEE increased degradability, total gas and CH4 production in all substrates (p < .05). Butyrate concentration also increased while acetate:propionate ratio and pH decreased with the addition of FEE (p < .05). These results indicated that the use of fibrolytic enzymes can be a reliable strategy to improve degradability of low-quality forages, contributing to the sustainability and intensification of livestock production in tropical countries.

Effects of Acacia mearnsii supplementation on nutrition, parasitological, blood parameters and methane emissions in Santa Inês sheep infected with Trichostrongylus colubriformis and Haemonchus contortus.

Gastrointestinal nematodes are a main problem for ruminant production, reducing animal performance and increasing environmental impact per unit of animal product generated. Tannin supplementation may lead to positive results regarding aspects such as parasitic infections and methane (CH4) emissions. Therefore, the objective of this experiment was to evaluate the effects of the condensed tannins (CT) extract made of powdered Acacia mearnsii bark (PAB) on nutrition, parasitic status and CH4 emissions in sheep artificially infected with Trichostrongylus colubriformis and Haemonchus contortus. Twenty 10-month old Santa Inês lambs (24.7 ± 3.14 kg of initial body weight) were used in a 50-day trial. Animals were divided in four treatment groups according to parasitic infection and PAB supplementation: two control groups without infections, one without PAB (C-) (n = 4) and one with PAB (C+) (n = 4); two infected groups, one without PAB (I-) (n = 6) and another receiving PAB (I+) (n = 6). Initially, animals were kept in individual pens where they were fed ad libitum chopped tifton 85 hay (Cynodon spp.) and 210 g/animal/day of concentrate. On the first day of experiment, animals of I- and I+ groups were artificially infected with infective larvae (L3) of T. colubriformis and H. contortus. Lambs were weighed fortnightly to calculate average daily body weight gain (ADG). Blood and faeces samples were also collected in the same moment of weighing for the evaluation of blood parameters and faecal egg count (FEC) respectively. After 40 days of experiment, measurements of CH4 emissions in small chamber system started and following that, apparent total tract digestibility (ATTD) assay was carried out in metabolic cages. In the end of experimental period (50 days), lambs were slaughtered and samples of abomasum and small intestine content were collected for worm count, identification, and eggs/female count. No significant (p > 0.05) treatment effects were verified for ADG, ATTD and worm count. Blood parameters were affected in both infected groups (p < 0.05) from the 28th experimental day onwards, when these animals started to show reduced red blood cells, haemoglobin and packed cell volume when compared to C- and C+. Decreased FEC was verified in I+ when compared to I- and also, H. contortus eggs/female worm for I+ was lower than for I- (p < 0.05). Both infected groups showed higher CH4 emissions than the control groups (p < 0.05). Results highlighted the anthelmintic potential of PAB and indicated methanogenic effect of parasitic nematode infections.

Methane emission of Santa Inês sheep fed cottonseed by-products containing different levels of gossypol.

The aim of this study was to evaluate the methane (CH4) emission of Santa Inês sheep fed cottonseed by-products, verifying if the gossypol content of these feedstuffs affects CH4 emission. Twelve late-lactating Santa Inês sheep (44.8 ± 7.5 kg body weight (BW)) were allocated in metabolic cages for an experimental period of 19 days, 14 days for adaptation and 5 days for measuring CH4 emission and dry matter intake (DMI). The animals were divided into four treatments, established in accordance with the cottonseed by-product used in concentrate formulation: Control (CON - no cottonseed by-product), Whole cottonseed (WCS), Cottonseed cake (CSC), and Cottonseed meal (CSM). The free gossypol level of the concentrates were 0, 1,276, 350, and 190 ppm for CON, WCS, CSC, and CSM, respectively. Also, the animals received Cynodon dactylon cv. Coast Cross hay, water, and mineral salt ad libitum. The ether extract content of the diets was balanced between treatments by including soybean oil in concentrates. The technique used to measure the CH4 emission was the sulfur hexafluoride (SF6) tracer technique, and the gas samples collected were quantified by analysis in gas chromatography system. The CH4 emission was evaluated considering the daily emission (g CH4/day); DMI (g CH4/kg DMI); and BW (g CH4/kg BW). No statistical difference was found (P > 0.05) between treatments for DMI and CH4 parameters. In the regression analysis, no significant relation (P > 0.05) between gossypol content and CH4 emission was observed. These results suggest that gossypol does not affect rumen methanogenesis.

Tropical tanniniferous legumes used as an option to mitigate sheep enteric methane emission.

This study presents the first results from Brazil using SF(6) tracer technique adapted from cattle to evaluate the capability of condensed tannin (CT) present in three tropical legume forages, Leucaena leucocephala (LEU), Styzolobium aterrimum (STA), and Mimosa caesalpiniaefolia Benth (MIM) to reduce enteric CH(4) production in Santa Inês sheep. Twelve male lambs [27.88 ± 2.85 kg body weight (BW)] were allocated in individual metabolic cages for 20-day adaptation followed by 6 days for measuring dry matter intake (DMI) and CH(4) emission. All lambs received water, mineral supplement, and Cynodon dactylon v. coast-cross hay ad libitum. The treatments consisted of soybean meal (710 g/kg) and ground corn (290 g/kg) [control (CON)]; soybean meal (150 g/kg), ground corn (30 g/kg), and Leucaena hay (820 g/kg) (LEU); soybean meal (160 g/kg), ground corn (150 g/kg), and Mucuna hay (690 g/kg) (STA); and soybean meal (280 g/kg), ground corn (190 g/kg), and Mimosa hay (530 g/kg) (MIM); all calculated to provide 40 g/kg CT (except for CON). DMI (in grams of DMI per kilogram BW per day) was lower for LEU (22.0) than CON (29.3), STA (31.2), and MIM (31.6). The LEU group showed emission of 7.8 g CH(4)/day, significantly lower than CON (10.5 g CH(4)/day), STA (10.4 g CH(4)/day), and MIM (11.3 g CH(4)/day). However, when the CH(4) emission per DMI was considered, there were no significant differences among treatments (0.37, 0.36, 0.33, and 0.35 g CH(4)/g DMI/kg BW/day, respectively, for CON, LEU, STA, and MIM). The sheep receiving STA had shown a tendency (p = 0.15) to reduce methane emission when compared to the CON group. Therefore, it is suggested that tropical tanniniferous legumes may have potential to reduce CH(4) emission in sheep, but more research is warranted to confirm these results.