Effects of fumaric acid supplementation on methane production and rumen fermentation in goats fed diets varying in forage and concentrate particle size

Background: In rumen fermentation, fumaric acid(FA) could competitively utilize hydrogen with methanogenesis to enhance propionate production and suppress methane emission, but both effects were diet-dependent. This study aimed to explore the effects of FA supplementation on methanogenesis and rumen fermentation in goats fed diets varying in forage and concentrate particle size.Methods: Four rumen-cannulated goats were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments: low or high ratio of forage particle size: concentrate particle size(Fps:Cps), without or with FA supplementation(24 g/d). Fps:Cps was higher in the diet with chopped alfalfa hay plus ground corn than in that with ground alfalfa hay plus crushed corn.Results: Both increasing dietary Fps:Cps and FA supplementation shifted ruminal volatile fatty acid(VFA) patterns toward more propionate and less acetate in goats. An interaction between dietary Fps:Cps and FA supplementation was observed for the ratio of acetate to propionate(A:P), which was more predominant when FA was supplemented in the low-Fps:Cps diet. Methane production was reduced by FA, and the reduction was larger in the low-Fps:Cps diet(31.72%) than in the high-Fps:Cps diet(17.91%). Fumaric acid decreased ruminal total VFA concentration and increased ruminal p H. No difference was found in ruminal DM degradation of concentrate or alfalfa hay by dietary Fps:Cps or FA. Goats presented a lower ruminal methanogen abundance with FA supplementation and a higher B. fibrisolvens abundance with high dietary Fps:Cps.Conclusions: Adjusting dietary Fps:Cps is an alternative dietary model for studying diet-dependent effects without changing dietary chemical composition. Fumaric acid supplementation in the low-Fps:Cps diet showed greater responses in methane mitigation and propionate increase.

High rumen degradable starch decreased goat milk fat via trans-10,cis-12 conjugated linoleic acid-mediated downregulation of lipogenesis genes,particularly,INSIG1

Background:Starch is an important substance that supplies energy to ruminants.To provide sufficient energy for high-yielding dairy ruminants,they are typically fed starch-enriched diets.However,starch-enriched diets have been proven to increase the risk of milk fat depression(MFD)in dairy cows.The starch present in ruminant diets could be divided into rumen-degradable starch(RDS)and rumen escaped starch(RES)according to their different degradation sites(rumen or intestine).Goats and cows have different sensitivities to MFD.Data regarding the potential roles of RDS in milk fat synthesis in the mammary tissue of dairy goats and in regulating the occurrence of MFD are limited.Results:Eighteen Guanzhong dairy goats(day in milk=185±12 d)with similar parity,weight,and milk yield were selected and randomly assigned to one of three groups(n=6),which were fed an LRDS diet(Low RDS=20.52%),MRDS diet(Medium RDS=22.15%),or HRDS diet(High RDS=24.88%)for 5 weeks.Compared with that of the LRDS group,the milk fat contents in the MRDS and HRDS groups significantly decreased.The yields of short-,mediumand long-chain fatty acids decreased in the HRDS group.Furthermore,increased RDS significantly decreased ruminal B.fibrisolvens and Pseudobutyrivibrio abundances and increased the trans-10,cis-12 conjugated linoleic acid(CLA)and trans-10 C18:1 contents in the rumen fluid.A multiomics study revealed that the HRDS diet affected mammary lipid metabolism down-regulation of ACSS2,MVD,AGPS,SCD5,FADS2,CERCAM,SC5D,HSD17B7,HSD17B12,ATM,TP53RK,GDF1 and LOC102177400.Remarkably,the significant decrease of INSIG1,whose expression was depressed by trans-10,cis-12 CLA,could reduce the activity of SREBP and,consequently,downregulate the downstream gene expression of SREBF1.Conclusions:HRDS-induced goat MFD resulted from the downregulation of genes involved in lipogenesis,particularly,INSIG1.Specifically,even though the total starch content and the concentrate-to-fiber ratio were the same as those of the high-RDS diet,the low and medium RDS diets did not cause MFD in lactating goats.

Dynamics of methanogenesis,ruminal fermentation and fiber digestibility in ruminants following elimination of protozoa: a meta-analysis

Background: Ruminal microbes are vital to the conversion of lignocellulose-rich plant materials into nutrients for ruminants.Although protozoa play a key role in linking ruminal microbial networks,the contribution of protozoa to rumen fermentation remains controversial; therefore,this meta-analysis was conducted to quantitatively summarize the temporal dynamics of methanogenesis,ruminal volatile fatty acid(VFA) profiles and dietary fiber digestibility in ruminants following the elimination of protozoa(also termed defaunation).A total of 49 studies from 22 publications were evaluated.Results: The results revealed that defaunation reduced methane production and shifted ruminal VFA profiles to consist of more propionate and less acetate and butyrate,but with a reduced total VFA concentration and decreased dietary fiber digestibility.However,these effects were diminished linearly,at different rates,with time during the first few weeks after defaunation,and eventually reached relative stability.The acetate to propionate ratio and methane production were increased at 7 and 11 wk after defaunation,respectively.Conclusions: Elimination of protozoa initially shifted the rumen fermentation toward the production of more propionate and less methane,but eventually toward the production of less propionate and more methane over time.

1,25-Dihydroxyvitamin D_3 modulates calcium transport in goat mammary epithelial cells in a dose- and energydependent manner

Background： Calcium is a vital mineral and an indispensable component of milk for ruminants. The regulation of transcellular calcium transport by 1,25-dihydroxyvitamin D3 （1,25-（OH）2D3, the active form of vitamin D） has been confirmed in humans and rodents, and regulators, including vitamin D receptor （VDR）, calcium binding protein Dgk （calbindin-Dgk）, plasma membrane Ca2＋-ATPase ] b （PMCAlb）, PMAC2b and Oral1, are involved in this process. However, it is still unclear whether 1,25-（OH）2D3 could stimulate calcium transport in the ruminant mammary gland. The present trials were conducted to study the effect of 1,25-（OH）2D3 supplementation and energy availability on the expression of genes and proteins related to calcium secretion in goat mammary epithelial cells. Methods： An in vitro culture method for goat secreting mammary epithelial cells was successfully established. The cells were treated with different doses of 1,25-（OH）2D3 （0, 0.1, 1.0, 10.0 and 100.0 nmol/L） for calcium transport research, followed by a 3-bromopyruvate （3-BrPA, an inhibitor of glucose metabolism） treatment to determine its dependence on glucose availability. Cell proliferation ratios, glucose consumption and enzyme activities were measured with commercial kits, and real-time quantitative polymerase chain reaction （RT-qPCR）, and western blots were used to determine the expression of genes and proteins associated with mammary calcium transport in dairy goats, respectively. Results： 1,25-（OH）2D3 promoted cell proliferation and the expression of genes involved in calcium transport in a dose-dependent manner when the concentration did not exceed 10.0 nmol/L. In addition, 100.0 nmol/L 1,25-（OH） 2D3 inhibited cell proliferation and the expression of associated genes compared with the 10.0 nmol/L treatment. The inhibition of hexokinase 2 （HK2）, a rate-limiting enzyme in glucose metabolism, decreased the expression of PMCA1 b and PMCA2b at the mRNA and protein levels as well as the transcription of Oral1, indicating that glucose avaitability was required for goat mammary calcium transport. The optimal concentration of 1,25-（OH）2D3 that facilitated calcium transport in this study was 10.0 nmol/L. Conclusions： Supplementation with 1,25-（OH）2D3 influenced cell proliferation and regulated the expression of calcium transport modulators in a dose- and energy-dependent manner, thereby highlighting the role of 1,25-（OH）2D3 as an efficacious regulatory agent that produces calcium-enriched milk in ruminants when a suitable energy status was guaranteed.

Potential roles of the rectum keystone microbiota in modulating the microbial community and growth performance in goat model

Background Ruminal microbiota in early life plays critical roles in the life-time health and productivity of ruminant animals.However,understanding of the relationship between gut microbiota and ruminant phenotypes is very limited.Here,the relationship between the rectum microbiota,their primary metabolites,and growth rate of a total of 76 young dairy goats(6-month-old)were analyzed,and then 10 goats with the highest or lowest growth rates respectively were further compared for the differences in the rectum microbiota,metabolites,and animal’s immune parameters,to investigate the potential mechanisms by which the rectum microbiota contributes to the health and growth rate.Results The analysis of Spearman correlation and microbial co-occurrence network indicated that some keystone rectum microbiota,including unclassified Prevotellaceae,Faecalibacterium and Succinivibrio,were the key modulators to shape the rectum microbiota and closely correlated with the rectum SCFA production and serum IgG,which contribute to the health and growth rate of young goats.In addition,random forest machine learning analysis suggested that six bacterial taxa in feces could be used as potential biomarkers for differentiating high or low growth rate goats,with 98.3%accuracy of prediction.Moreover,the rectum microbiota played more important roles in gut fermentation in early life(6-month-old)than in adulthood stage(19-month-old)of goats.Conclusion We concluded that the rectum microbiota was associated with the health and growth rate of young goats,and can be a focus on the design of the early-life gut microbial intervention.

Yak rumen microbiome elevates fiber degradation ability and alters rumen fermentation pattern to increase feed efficiency

Rumen microbes play an important role in ruminant energy supply and animal performance.Previous studies showed that yak(Bos grunniens)rumen microbiome and fermentation differ from other ruminants.However,little is understood about the features of the rumen microbiome that make yak adapted to their unique environmental and dietary conditions.This study was to investigate the rumen microbiome and metabolome to understand how yak adapt to the coarse forage and harsh environment in the Qinghai-Tibetan plateau.Nine female Qaidam yellow cattle(Bos taurus),9 dzomo(hybrids of cattle and yak)and 9 female plateau yak(B.grunniens),about 5 to 6 years old,were used in this study.Rumen fermentation parameters,fibrolytic enzyme activities,and rumen metataxonomic were determined.Then 18(6 samples per group)were selected for rumen metagenomic and metabolome analysis.Metataxonomic analysis revealed that the rumen microbiota was significantly different among plateau yak,Qaidam yellow cattle,and dzomo(P<0.05).Metagenomic analysis displayed a larger gene pool encoding a richer repertoire of carbohydrate-active enzymes in the rumen microbiome of plateau yak and dzomo than Qaidam yellow cattle(P<0.05).Some of the genes encoding glycoside hydrolases that mediate the digestion of cellulose and hemicellulose were significantly enriched in the rumen of plateau yak than Qaidam yellow cattle,but glycoside hydrolase 57 that primarily includes amylases was abundant in Qaidam yellow cattle(P<0.05).The rumen fermentation profile differed also,Qaidam yellow cattle having a higher molar proportion of acetate but a lower molar proportion of propionate than dzomo and plateau yak(P<0.05).Based on metabolomic analysis,rumen microbial metabolic pathways and metabolites were different.Differential metabolites are mainly amino acids,carboxylic acids,sugars,and bile acids.Changes in rumen microbial composition could explain the above results.The present study showed that the rumen microbiome of plateau yak helps its host to adapt to the Qinghai-Tibetan plateau.In particular,the plateau yak rumen microbiome has more enzymes genes involved in cellulase and hemicellulase than that of cattle,resulting higher fibrolytic enzyme activities inyak,further providing stronger fiber degradation function.

Regulation of pancreatic exocrine in ruminants and the related mechanism: The signal transduction and more

The unique structure of the stomach,including the rumen,reticulum,omasum,and abomasum,indicates the differences between the ruminant and monogastric animals in the digestion of nutrients.This difference is reflected in the majority of dietary nutrients that may be fermented in the rumen.Significant proteins and a certain amount of starch can flow to the small intestine apart from rumen.The initial phase of small intestinal digestion requires pancreatic digestive enzymes.In theory,the enzymatic digestion and utilization efficiency of starch in the small intestine are considerably higher than that in the rumen,but the starch digestibility in the small intestine is quite low in ruminants.Therefore,improving the digestion of nutrients,especially starch in the small intestine is more urgent for high-yield ruminants.Although the pancreas plays a central role in nutrient digestion,the progress of research investigating pancreatic exocrine regulation in the ruminant is slow due to some factors,such as the complex structure of the pancreas,the selection of experimental model and duration,and internal(hormones or ages) and external(diet) influences.The present review is based on the research findings of pancreatic exocrine regulation of dairy animals and expounded from the physiological structure of the ruminant pancreas,the factors affecting the digestion and exocrine processing of carbohydrates,and the regulatory mechanism governing this process.The review aims to better understand the characteristics of enzymatic digestion,thereby advancing pancreatic exocrine research and improving the digestion and utilization of nutrients in ruminants.Additionally,this review provides the theoretical basis for improving nutrient utilization efficiency,reducing wastage of feed resources,and promoting the efficient development of the dairy industry.

Rumen bacterial cluster identification and its influence on rumen metabolites and growth performance of young goats

Enterotypes,which are defined as bacterial clusters in the gut microbiome,have been found to have a close relationship to host metabolism and health.However,this concept has never been used in the rumen,and little is known about the complex biological relationships between ruminants and their rumen bacterial clusters.In this study,we used young goats(n=99)as a model,fed them the same diet,and analyzed their rumen microbiome and corresponding bacterial clusters.The relationships between the bacterial clusters and rumen fermentation and growth performance in the goats were further investigated.Two bacterial clusters were identified in all goats:the P-cluster(dominated by genus Prevotella,n=38)and R-cluster(dominated by Ruminococcus,n=61).Compared with P-cluster goats,Rcluster goats had greater growth rates,concentrations of propionate,butyrate,and 18 free amino acids)and proportion of unsaturated fatty acids,but lower acetate molar percentage,acetate to propionate ratio,and several odd and branched chain and saturated fatty acids in rumen fluid(P<0.05).Several members of Firmicutes,including Ruminococcus,Oscillospiraceae NK4A214 group,and Christensenellaceae R-7 group were significantly higher in the R-cluster,whereas Prevotellaceae members,such as Prevotella and Prevotellaceae UCG-003,were significantly higher in P-cluster(P<0.01).Co-occurrence networks showed that R-cluster enriched bacteria had significant negative correlations with P-cluster enriched bacteria(P<0.05).Moreover,we found the concentrations of propionate,butyrate and free amino acids,and the proportions of unsaturated fatty acids were positively correlated with R-cluster enriched bacteria(P<0.05).The concentrations of acetate,acetate to propionate ratio,and the proportion of odd and branched chain and saturated fatty acids were positively correlated with P-cluster enriched bacteria(P<0.05).Overall,our results indicated that rumen bacterial clusters can influence rumen fermentation and growth performance of young goats,which may shed light on modulating the rumen microbiome in early life to improve the growth performance of ruminant animals.