Winery by-products as a feed source with functional properties: dose-response effect of grape pomace, grape seed meal, and grape seed extract on rumen microbial community and their fermentation activity in RUSITEC.

BACKGROUND: Grape and winery by-products have nutritional values for cattle and also contain functional compounds like phenols, which not only bind to protein but can also directly affect microbiota and their function in the rumen. We characterized the nutritional and functional effects of grape seed meal and grape pomace as well as an effective dosage of grape phenols on ruminal microbiota and fermentation characteristics using a rumen simulation technique. RESULTS: Six diets (each n = 8) were compared including a control diet (CON, no by-product), a positive control diet (EXT, CON + 3.7% grape seed extract on a dry matter (DM) basis), two diets with grape seed meal at 5% (GS-low) and 10% (GS-high), and two diets with grape pomace: at 10% (GP-low) and 20% (GP-high), on a DM basis. The inclusion of the by-product supplied total phenols at 3.4%, 0.7%, 1.4%, 1.3%, and 2.7% of diet DM for EXT, GS-low, GS-high, GP-low, and GP-high, respectively. Diets were tested in four experimental runs. All treatments decreased ammonia concentrations and the disappearances of DM and OM (P < 0.05) compared to CON. EXT and GP-high lowered butyrate and odd- and branch-chain short-chain fatty acids while increased acetate compared to CON (P < 0.05). Treatments did not affect methane formation. EXT decreased the abundance of many bacterial genera including those belonging to the core microbiota. GP-high and EXT consistently decreased Olsenella and Anaerotipes while increased Ruminobacter abundances. CONCLUSION: The data suggest that the inclusion of winery by-products or grape seed extract could be an option for reducing excessive ammonia production. Exposure to grape phenols at a high dosage in an extract form can alter the rumen microbial community. This, however, does not necessarily alter the effect of grape phenols on the microbial community function compared to feeding high levels of winery by-products. This suggests the dominant role of dosage over the form or source of the grape phenols in affecting ruminal microbial activity. In conclusion, supplementing grape phenols at about 3% of diet DM is an effective dosage tolerable to ruminal microbiota.

Influence of Direct-fed Microbial Blend and Ferula Elaeochytris on in Vitro Rumen Fermentation Pattern and Degradability During Simulated Ruminal Acidosis.

Introduction: The use of probiotics and phytobiotics has attracted interest because of their protective effect against acidosis. Ferula elaeochytris (FE) is considered a good source of bioactive compounds, mainly monoterpene α-pinene. This study aimed to investigate the effect of a direct-fed microbial blend (Pro) and FE on rumen fermentation parameters in vitro under normal and acidosis conditions. Material and Methods: An in vitro experiment using the Hohenheimer Futterwerttest (HFT) gas production system was conducted. An acidosis challenge was made to compare the effectiveness of the probiotics blend and FE extract on ruminal pH regulation. To generate different ruminal fermentation parameters, the design of the trial considered the 2 additives (Pro and FE) × 6 incubation times (2, 4, 8, 12, 24 and 48 h) × 2 conditions (acidosis and normal) × 2 incubation runs for each feedstuff (barley, alfalfa and straw). Results: An acidosis challenge was successfully induced. The Pro and FE additives had no impact on the observed rumen fermentation parameters such as volatile fatty acid concentration or ammonia (P = 0.001). The acidosis condition decreased total in vitro degradability (IVD) by 3.5% and 21.9% for barley and straw, respectively (P < 0.001). The additives had different significant effects on the IVD of nutrients during both normal and acidosis conditions. In alfalfa samples, FE supplementation significantly decreased the IVD of all observed nutrients under the ruminal acidosis condition, although it had no effect during the normal condition. Conclusion: An acidosis challenge was successfully induced and the effect of additives was varied on fermentation parameters and rumen degradability of different feeds either under normal or acidosis conditions.

Ameliorative Effect of Oxytocin on FBN1 and PEPCK Gene Expression, and Behavioral Patterns in Rats' Obesity-Induced Diabetes.

Amelioration of hyperinsulinemia and insulin resistance associated with obesity is a cardinal target for therapeutics. Therefore, we investigated the relation of Fibrilln-1 (FBN1) mRNA expression and hepatic phosphoenolpyruvate caboxykinase (PEPCK) enzyme to the ameliorative impact of oxytocin on obesity-induced diabetes, suggesting glycogenolysis markers in diabetic models. Four groups of forty male Wistar rats were formed (n = 10): a control group fed basal diet and intraperitoneal injections of saline; an oxytocin-injected group; a diet-induced obese group fed a high-fat/high-sugar diet and injected with saline; a diet-induced obese group injected with oxytocin. Depending on blood glucose levels, obese groups were further sub-grouped into prediabetic, and diabetic rats, with 5 rats each, at the ninth and the 16th week of the feeding period, respectively. FBN1 expression and PEPCK activity were determined using the qPCR technique and some biochemical parameters (glycemic, lipid profile, kidney, and liver functions) were determined using kits. Obese groups showed an elevation of brain FBN1 expression, high serum lipid profile, high glucose level, and a deleterious impact on liver and kidney functions. Obese groups showed the stimulator effect of the PEPCK enzyme and time-dependent pathological changes in renal and hepatic tissues. The motor activities were negatively correlated with FBN1 gene expression in prediabetic and diabetic rats. In addition to our previous review of the crucial role of asprosin, here we showed that oxytocin could ameliorate obesity-induced diabetes and decrease FBN1 gene expression centrally to block appetite. Oxytocin caused decreases in PEPCK enzyme activity as well as glycogenolysis in the liver. Therefore, oxytocin has a potential effect on FBN1 expression and PEPCK enzyme activity in the obesity-induced diabetic-rat model.

Unveiling the Bovine Epimural Microbiota Composition and Putative Function.

Numerous studies have used the 16S rRNA gene target in an attempt to characterize the structure and composition of the epimural microbiota in cattle. However, comparisons between studies are challenging, as the results show large variations associated with experimental protocols and bioinformatics methodologies. Here, we present a meta-analysis of the rumen epimural microbiota from 11 publicly available amplicon studies to assess key technical and biological sources of variation between experiments. Using the QIIME2 pipeline, 332 rumen epithelial microbiota samples were analyzed to investigate community structure, composition, and functional potential. Despite having a significant impact on microbial abundance, country of origin, farm, hypervariable region, primer set, animal variability, and biopsy location did not obscure the identification of a core microbiota. The bacterial genera Campylobacter, Christensenellaceae R-7 group, Defluviitaleaceae UCG-011, Lachnospiraceae UCG-010, Ruminococcaceae NK4A214 group, Ruminococcaceae UCG-010, Ruminococcaceae UCG-014, Succiniclasticum, Desulfobulbus, and Comamonas spp. were found in nearly all epithelium samples (>90%). Predictive analysis (PICRUSt) was used to assess the potential functions of the epithelial microbiota. Regularized canonical correlation analysis identified several pathways associated with the biosynthesis of precursor metabolites in Campylobacter, Comamonas, Desulfobulbus, and Ruminococcaceae NK4A214, highlighting key metabolic functions of these microbes within the epithelium.

Effects of the supplementation of plant-based formulations on microbial fermentation and predicted metabolic function in vitro.

This study aimed at testing the effects of three different formulations of feed supplements based on three different combinations of plant derived alkaloids, prebiotics, tannins, vitamins and minerals on rumen fermentation and the microbiome in vitro. A Rusitec experiment was conducted in 2 identical runs using a complete randomized design with 3 replicates per treatment resulting in total of 6 treatment combinations (n = 6). Each run lasted 12 d with sampling occurring in the last 5 d. Diets were a standard dairy ration (60:40; concentrate:forage) supplemented with one of 3 different plant-based combinations (PI, PII, and PIII) at a level of 100 mg/l and a non-supplemented control (basal diet, control). Microbial DNA samples were taken on the last day of each run and the 16S rRNA target gene sequenced using Illumina MiSeq technology. The supplementations had no effect on the pH, methane and carbon dioxide production. However, both total SCFA (P = 0.08) and molar concentrations of acetate (P = 0.06) tended to be increased in the treatment groups in comparison to control, with PII having the highest overall values (102.7 mmol/L and 43.3 mmol/L, respectively). Alpha diversity indices Shannon, Simpson and Chao1 showed no effect of supplementations or combinations. The addition of PII increased the relative abundance of Bacteroidetes compared to all other treatments (P = 0.05). Supplementation with plant-based combinations reduced the relative abundance of Pyramidobacter from the family Dethiosulfovibrionaceae in comparison with the control diet (P = 0.05). Evaluation of predicted gene function through PICRUSt analysis showed variation in predicted cellular function and metabolism between bacterial communities supplemented with plant-based combinations compared to the control diet. This shows that the addition of plant-based combinations can have the potential to modulate the metabolic function of rumen microbes, and likely the production of small-sized rumen metabolites, without disrupting the rumen microbial community structure and diversity.

Rumen microbial abundance and fermentation profile during severe subacute ruminal acidosis and its modulation by plant derived alkaloids in vitro.

Rumen microbiota have important metabolic functions for the host animal. This study aimed at characterizing changes in rumen microbial abundances and fermentation profiles using a severe subacute ruminal acidosis (SARA) in vitro model, and to evaluate a potential modulatory role of plant derived alkaloids (PDA), containing quaternary benzophenanthridine and protopine alkaloids, of which sanguinarine and chelerythrine were the major bioactive compounds. Induction of severe SARA strongly affected the rumen microbial composition and fermentation variables without suppressing the abundance of total bacteria. Protozoa and fungi were more sensitive to the low ruminal pH condition than bacteria. Induction of severe SARA clearly depressed degradation of fiber (P < 0.001), which came along with a decreased relative abundance of fibrolytic Ruminococcus albus and Fibrobacter succinogenes (P < 0.001). Under severe SARA conditions, the genus Prevotella, Lactobacillus group, Megasphaera elsdenii, and Entodinium spp. (P < 0.001) were more abundant, whereas Ruminobacter amylophilus was less abundant. SARA largely suppressed methane formation (-70%, P < 0.001), although total methanogenic 16S rRNA gene abundance was not affected. According to principal component analysis, Methanobrevibacter spp. correlated to methane concentration. Addition of PDA modulated ruminal fermentation under normal conditions such as enhanced (P < 0.05) concentration of total SCFA, propionate and valerate, and increased (P < 0.05) degradation of crude protein compared with the unsupplemented control diet. Our results indicate strong shifts in the microbial community during severe SARA compared to normal conditions. Supplementation of PDA positively modulates ruminal fermentation under normal ruminal pH conditions.