Evaluation of Ruminal Dry Matter Disappearance and pH of Dry Corn, High-Moisture Corn, and Rye under in Vitro Conditions

An in vitro experiment was conducted to evaluate the effects of grain type on in vitro dry matter disappearance (IVDMD) and pH using ruminal fluid and a buffer reagent. Five beef cows were used for ruminal fluid collection and ruminal fluid was pooled prior to use. The cows used for ruminal fluid collection were maintained on a forage-based diet (60:40 forage to concentrate) for 28 d prior to ruminal fluid collection. Three grain types were evaluated: 1) dry corn (89% dry matter;DRC), 2) high-moisture corn (72% dry matter;HMC) and 3) rye (89% dry matter;RYE). Data were analyzed as a completely randomized design. A total of twenty-one replications of each grain type were used for statistical analysis (n = 63 tubes total). IVDMD was greater (P ≤ 0.05) by 97.1% and 46.4% for RYE compared to DRC and HMC, respectively. Additionally, IVDMD was greater (P = 0.05) by 34.6% for HMC compared to DRC. Final pH was decreased (P ≤ 0.05) for RYE by 12.3% and 2.8% for RYE compared to DRC and HMC, respectively. Also, final pH was decreased (P = 0.05) by 9.8% for HMC compared to DRC. These data indicate that corn harvest method (dry vs. high-moisture) influences IVDMD and final pH and that rye has a greater disappearance of dry matter and lower final pH than corn under in vitro conditions.

Susceptibility of dairy cows to subacute ruminal acidosis is reflected in both prepartum and postpartum bacteria as well as odd-and branched-chain fatty acids in feces

Background:The transition period is a challenging period for high-producing dairy cattle.Cows in early lactation are considered as a group at risk of subacute ruminal acidosis(SARA).Variability in SARA susceptibility in early lactation is hypothesized to be reflected in fecal characteristics such as fecal pH,dry matter content,volatile and odd-and branched-chain fatty acids(VFA and OBCFA,respectively),as well as fecal microbiota.This was investigated with 38 periparturient dairy cows,which were classified into four groups differing in median and mean time of reticular pH below 6 as well as area under the curve of pH below 6.Furthermore,we investigated whether fecal differences were already obvious during a period prior to the SARA risk(prepartum).Results:Variation in reticular pH during a 3-week postpartum period was not associated with differences in fecal pH and VFA concentration.In the postpartum period,the copy number of fecal bacteria and methanogens of unsusceptible(UN)cows was higher than moderately susceptible(MS)or susceptible(SU)cows,while the genera Ruminococcus and Prevotellacea_UCG-001 were proportionally less abundant in UN compared with SU cows.Nevertheless,only a minor reduction was observed in iso-BCFA proportions in fecal fatty acids of SU cows,particularly iso-C15:0and iso-C16:0,compared with UN cows.Consistent with the bacterial changes postpartum,the lower abundance of Ruminococcus was already observed in the prepartum fecal bacterial communities of UN cows,whereas Lachnospiraceae_UCG-001 was increased.Nevertheless,no differences were observed in the prepartum fecal VFA or OBCFA profiles among the groups.Prepartum fecal bacterial communities of cows were clustered into two distinct clusters with 70%of the SU cows belonging to cluster 1,in which they represented 60%of the animals.Conclusions:Inter-animal variation in postpartum SARA susceptibility was reflected in post-and prepartum fecal bacterial communities.Differences in prepartum fecal bacterial communities could alert for susceptibility to develop SARA postpartum.Our results generated knowledge on the association between fecal bacteria and SARA development which could be further explored in a prevention strategy.

Coordinated response of milk bacterial and metabolic profiles to subacute ruminal acidosis in lactating dairy cows

Background Bovine milk is an important source of nutrition for human consumption,and its quality is closely associated with the microbiota and metabolites in it.But there is limited knowledge about the milk microbiome and metabolome in cows with subacute ruminal acidosis.Methods Eight ruminally cannulated Holstein cows in mid lactation were selected for a 3-week experiment.The cows were randomly allocated into 2 groups,fed either a conventional diet(CON;40%concentrate;dry matter basis)or a high-concentrate diet(HC;60%concentrate;dry matter basis).Results The results showed that there was a decreased milk fat percentage in the HC group compared to the CON group.The amplicon sequencing results indicated that the alpha diversity indices were not affected by the HC feeding.At the phylum level,the milk bacteria were dominated by Proteobacteria,Actinobacteria,Bacteroidetes,and Firmicutes both in the CON and HC groups.At the genus level,the HC cows displayed an improved proportion of Labrys(P=0.015)compared with the CON cows.Results of both the principal components analysis and partial least squares of discriminant analysis of milk metabolome revealed that samples of the CON and HC groups clustered separately.A total of 31 differential metabolites were identified between the two groups.Of these,the levels of 11 metabolites decreased(α-linolenic acid,prostaglandin E2,L-lactic acid,L-malic acid,3-hydroxysebacic acid,succinyladenosine,guanosine,pyridoxal,L-glutamic acid,hippuric acid,and trigonelline),whereas the levels of the other 20 metabolites increased in the HC group with respect to the CON group(P<0.05).Conclusion These results suggested that subacute ruminal acidosis less impacted the diversity and composition of milk microbiota,but altered the milk metabolic profiles,which led to the decline of the milk quality.

Biochar’s Electrochemical Properties Impact on Methanogenesis: Ruminal vs. Soil Processes

The chemical composition of biochar and the pyrolysis temperature, under which biochar was produced, determine its electrochemical properties. Electrical conductivity, pseudo-capacitance, and double layer capacitance are the three main electrochemical properties of biochar. Due to the electrical conductivity biochar is able to interfere with the electrons flow and play a dual role of an electron donor or an electron acceptor. The average conductivity of biochar is 229.20 S/m. Pseudocapacitance of biochar lets it serve as a hydrogen sink, taking up the hydrogen produced by protozoa and preventing it from participating in methane-producing reactions in the rumen environment. The average value of biochar’s pseudocapacitance is 228 F·g-1. Positive and negative charges get stored due to the absorption of ions onto the carbon surface, which happens because of the existence of double layer capacitance as one of biochar’s electrochemical properties. Biochar’s double layer capacitance values can reach the point of 110.8 F·g-1. The electrochemical properties of biochar are directly co-dependent with its redox potential and pH. Electrical conductivity, pseudocapacitance, and double layer capacitance can significantly influence biochemical processes in the rumen and, thus, need to be studied practically.

Residual feed intake in beef cattle and its association with carcass traits, ruminal solid-fraction bacteria, and epithelium gene expression

Background: Residual feed intake(RFI) describes an animal’s feed efficiency independent of growth performance.The objective of this study was to determine differences in growth performance, carcass traits, major bacteria attached to ruminal solids-fraction, and ruminal epithelium gene expression between the most-efficient and the least-efficient beef cattle. One-hundred and forty-nine Red Angus cattle were allocated to three contemporary groups according to sex and herd origin. Animals were fed a finishing diet in confinement for 70 d to determine the RFI category for each. Within each group, the two most-efficient(n = 6; RFI coefficient =-2.69 ± 0.58 kg dry matter intake(DMI)/d) and the two least-efficient animals(n = 6; RFI coefficient = 3.08 ± 0.55 kg DMI/d) were selected. Immediately after slaughter, ruminal solids-fraction and ruminal epithelium were collected for bacteria relative abundance and epithelial gene expression analyses, respectively, using real-time PCR.Results: The most-efficient animals consumed less feed(P = 0.01; 5.03 kg less DMI/d) compared with the leastefficient animals. No differences(P > 0.10) in initial body weight(BW), final BW, and average daily gain(ADG) were observed between the two RFI classes. There were no significant RFI × sex effects(P > 0.10) on growth performance.Compared with the least-efficient group, hot carcass weight(HCW), ribeye area(REA), and kidney, pelvic, and heart fat(KPH) were greater(P ≤ 0.05) in the most-efficient cattle. No RFI × sex effect(P > 0.10) for carcass traits was detected between RFI groups. Of the 10 bacterial species evaluated, the most-efficient compared with least efficient cattle had greater(P ≤ 0.05) relative abundance of Eubacterium ruminantium, Fibrobacter succinogenes, and Megasphaera elsdenii, and lower(P ≤ 0.05) Succinimonas amylolytica and total bacterial density. No RFI × sex effect on ruminal bacteria was detected between RFI groups. Of the 34 genes evaluated in ruminal epithelium, the mostefficient cattle had greater(P ≤ 0.05) abundance of genes involved in VFA absorption, metabolism, ketogenesis, and immune/inflammation-response. The RFI × sex interactions indicated that responses in gene expression between RFI groups were due to differences in sex. Steers in the most-efficient compared with least-efficient group had greater(P ≤ 0.05) expression of SLC9 A1, HIF1 A, and ACO2. The most-efficient compared with least-efficient heifers had greater(P ≤ 0.05) m RNA expression of BDH1 and lower expression(P ≤ 0.05) of SLC9 A2 and PDHA1.Conclusions: The present study revealed that greater feed efficiency in beef cattle is associated with differences in bacterial species and transcriptional adaptations in the ruminal epithelium that might enhance nutrient delivery and utilization by tissues. The lack of RFI × sex interaction for growth performance and carcass traits indicates that sex may not play a major role in improving these phenotypes in superior RFI beef cattle. However, it is important to note that this result should not be considered a solid biomarker of efficient beef cattle prior to further examination due to the limited number of heifers compared with steers used in the study.

High-concentrate feeding upregulates the expression of inflammation-related genes in the ruminal epithelium of dairy cattle

Background： The objective of this study was to characterize the mRNA expression profile related to rumen epithelial inflammation through the in vivo and in vitro experiments. In the in vivo experiment, rumen papillae were collected from four dairy cows adapted to either a 40 % （LC） or 70 % （HC） concentrate feeds for microarray analysis. Results： Results showed that 245 differentially expressed genes （DEGs） were detected in the cows fed the HC relative to the LC diet. The DEGs were first annotated, and results revealed that the expression of inflammation- related genes, including IL-1t8, 1L-2, IL-22, CCL19, CCLS, CX3CR1, CXCL6, INHBE, LEPR, PRL, and TNFRSF9 found in the cytokine-cytokine receptor pathway were up-regulated in the HC-fed cows, indicating local inflammation in the rumen epithelium was triggered. The expression of IL-1~, 1l_-2, and IL-6 was further validated by qRT-PCR. To demonstrate whether there were relationships between cytokine mRNA expression and ruminal factors （pH and LPS）, the isolated ruminal epithelial cells were cultured in vitro. Results showed that the mRNA expression of IL-1, IL-2, IL-6, and IL-8 increased after the LPS treatment, while Iow-pH treatment elevated the mRNA expression of TNF-a, suggesting that Iow-pH coupled with higher levels of LPS in rumen of cows fed the HC may be mainly responsible for the triggered local ruminal inflammation. Conclusions： Our results indicate that ruminal local inflammation response might be triggered during HC feeding and these findings also enhance the knowledge of rumen epithelial adaptation to HC at the molecular level.

Changes in feed intake, nutrient digestion, plasma metabolites, and oxidative stress parameters in dairy cows with subacute ruminal acidosis and its regulation with pelleted beet pulp

The objectives of this study were to 1） determine the variation of nutrient digestion, plasma metabolites and oxidative stress parameters triggered by induced subacute ruminal acidosis （SARA）; and 2） evaluate the ability of pelleted beet pulp （BP） as a replacement for ground corn to alleviate SARA. Eight Holstein-Friesian cows were fed four diets during four successive17 day periods： 1） total mixed ration （TMR） containing 0% finely ground wheat （FGW） （WO）; 2） TMR containing 10% FGW （W10）; 3） TMR containing 20% FGW （W20）; and 4） TMR containing 10% BP as a replacement for 10% ground corn （BP10）. The SARA induction protocol reduced the mean ruminal pH from 6.37 to 5.94, and the minimum ruminal pH decreased from 5.99 to 5.41 from baseline to challenge period. Mean ruminal pH increased from 5.94 to 6.05, and minimum daily ruminal pH increased from 5.41 to 5.63, when BP was substituted for corn. The apparent digestibility of nutrients was not affected by the dietary treatments, except that the digestibility of neutral detergent fibre （NDF） and acid detergent fibre （ADF） was reduced in cows fed the W20 diet compared with cows fed the W0 and W10 diets, and cows fed the BP10 diet had higher NDF and ADF digestibility than the cows fed the W20 diet. Cows fed the W20 diet had a lower plasma concentration of 13-hydroxybutyrate （BHBA）, non-esterified fatty acids （NEFA）, cholesterol, triglyceride, and total antioxidative capacity （TAC）, and a higher plasma concentration of glucose, insulin, malonaldehyde （MDA）, super oxygen dehydrogenises （SOD）, and glutathione peroxidase （GSH-Px） than cows fed the W0 diet. Substitution of BP for corn increased concentrations of plasma BHBA and TAC, but decreased concentrations of plasma MDA. Our results indicate that reduction of fibre digestion; the concomitant increase of plasma glucose and insulin; the decrease of plasma BHBA, NEFA, cholesterol, and triglyceride; and changes of plasma oxidative stress parameters are highly related to SARA induced by W20 diets. These variables may be alternative candidates for SARA diagnosis. We also suggest that the substitution of BP for corn could reduce the risk of SARA, increase fibre digestion, and improve the antioxidant status in dairy cows.

Effects of Neutral Detergent Soluble Fiber and Sucrose Supplementation on Ruminal Fermentation, Microbial Synthesis, and Populations of Ruminal Cellulolytic Bacteria Using the Rumen Simulation Technique (RUSITEC)

We evaluated the effects of neutral detergent soluble fiber （NDSF） and sucrose supplementation on ruminal fermentation, microbial synthesis, and populations of ruminal cellulolytic bacteria using the rumen simulation technique （RUSITEC）. The experiment had a 2x2 factorial design with two dosages of sucrose, low （ca. 0.26 g d-1, low-sucrose） and high （ca. 1.01 g d-1, high-sucrose）, and two dosages of supplied NDSF, low （1.95 g d-1, Iow-NDSF） and high （2.70 g d-1, high-NDSF）. Interactions between NDSF and sucrose were detected for xylanase activity from solid fraction and apparent disappearance of neutral detergent fiber （NDF） and hemicellulose, with the lowest values observed for high-NDSF and high-sucrose treatment. Supplemental NDSF appeared to increase the molar proportion of acetate and reduce that of butyrate; however, the effects of supplemental sucrose on VFA profiles depended upon NDSF amount. There was a NDSFxsucrose interaction for the production of methane. High-NDSF fermenters had lower ammonia-N production, greater daily N flow of solid- associated microbial pellets and total microorganisms, and greater microbial synthesis efficiency compared with low- NDSF fermenters. Supplementation with NDSF resulted in an increase in 16S rDNA copies of Ruminococcusflavefaciens and a reduction in copies of Ruminococcus albus. Supplementation with sucrose tended to increase the 16S rDNA copies ofR. albus from liquid fraction, but did not affect daily total microbial N flow and cellulolytic bacterium populations from solid fraction. These data indicate that the effects of the interaction between NDSF and sugars on ruminal fermentation and fiber digestion should be taken into account in diet formulation. Ruminal fermentation and metabolism of sugars warrant further investigation.

Screening and characterization of a novel ruminal cellulase gene(Umcel-1) from a metagenomic library of gayal(Bos frontalis)

Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments. The aim of this study was to identify specific cellulase in the gayal rumen. A metagenomic fosmid library was constructed using genomic DNA isolated from the ruminal contents of four adult gayals. This library contained 38400 clones with an average insert size of 35.5 kb. The Umcel-1 gene was isolated from this library. Investigation of the cellulase activity of 24 random clones led to the identification of the Umcel-1 gene, which exhibited the most potent cellulase activity. Sequencing the Umcel-1 gene revealed that it contained an open reading frame of 942 base pairs that encoded a product of 313 amino acids. The putative gene Umcel-1 product belonged to the glycosyl hydrolase family 5 and showed the highest homology to the cellulase （GenBank accession no. YP_004310852.1 ） from Clostridium lentocellum DSM 5427, with 44% identity and 62% similarity. The Umcel-1 gene was heterologously expressed in Escherichia coil BL21, and recombinant Umcel-1 was purified. The activity of purified recombinant Umcel-1 was assessed, and the results revealed that it hydrolyzed carboxymethyl cellulose with optimal activity at pH 5.5 and 45~C. To our knowledge, this study provides the first evidence for a cellulase produced by bacteria in gayal rumen.

Effects of Molasses-Urea Supplementation on Weight Gain,Ruminal Fermentation and Major Microbe Populations of Winter-Grazing Sheep in Inner Mongolia

The present experiment was conducted to determine the effects of Molasses-Urea Supplementation （MUS） on weight gain, ruminal fermentation and major microbial populations in sheep on a winter grazing regime in Inner Mongolia. Total 40 sheep, allowed free consumption of MUS after grazing, served as a treatment group, while 30 sheep, fed only by pasture grazing, served as a control group. Ruminal fermentation parameters, consisted of pH, Bacterial Crude Protein （BCP） and ammonia nitrogen （NH3-N） were measured. In addition, numbers of five symbiotic bacteria were investigated. The results showed as follows： the average daily weight gain, concentration of NH3-N and numbers of protozoa were significantly higher （p〈0.05） in the treatment group than those in the control group. Contrastingly, no significant difference was found in BCP concentration and pH between the two groups. At the end of the experiment, the populations of Selenomonas ruminantium, Anaerovibrio lipolytica, Fibrobacter succinogenes, Ruminococcus flaveciens and Ruminococcus albus in the treatment group were significantly higher than those of the control group （p〈0.05）. These results demonstrated that greater weight gain could be induced during winter in Inner Mongolia by improved nutritional status through promotion of microbial populations using urea and sugar.

Effects of dietary forage to concentrate ratio and wildrye length on nutrient intake, digestibility, plasma metabolites, ruminal fermentation and fecal microflora of male Chinese Holstein calves

Twenty-eight male, weaned Chinese Holstein calves（（156.8±33.4） kg） were used to investigate the effects of dietary forage to concentrate ratio（F：C） and forage length on nutrient digestibility, plasma metabolites, ruminal fermentation, and fecal microflora. Animals were randomly allocated to four treatments in a 2×2 factorial arrangement： whole-length forage（WL） with low F：C（50：50）; WL with high F：C（65：35）; short-length forage（SL） with high F：C（65：35）; and SL with low F：C（50：50）. Chinese wildrye was used as the only forage source in this trial. The grass in the SL treatments was chopped using a chaff cutter to achieve small particle size（-50% particles 〉19 mm）. Dry matter intake（DMI） and organic matter（OM） intake was increased by increasing both F：C（P〈0.01） and forage length（FL）（P〈0.05）, while acid detergent fiber（ADF） and neutral detergent fiber（NDF） intakes were only increased by increasing the F：C（P〈0.01）. The digestibility of NDF was increased as the FL increased（P〈0.01）, and it was also affected by interaction between F：C and FL（P〈0.05）. Cholesterol（CHO）（P〈0.01）, leptin（LP）（P〈0.05）, and growth hormone（GH）（P〈0.01） concentrations in plasma were increased as dietary F：C increased. A significant increase in plasma triglyceride（TG）（P〈0.01）, insulin（INS）（P〈0.05）, and GH（P〈0.01） levels was observed with decreasing dietary FL. Ruminal p H values of calves fed with low F：C diets were significantly lower than those in high F：C treatment（P〈0.05）. Increasing the F：C enhanced ruminal acetic acid（P〈0.05） and acetic acid/propionic acid（P〈0.01）. Fecal Lactobacillus content was significantly higher, while Escherichia coli and Salmonella contents were significantly lower in WL and high F：C groups（P〈0.05）. Lower fecal scores（higher diarrhea rate） were observed in calves fed with SL hay compared to WL hay（P〈0.05）. Denatured gradient gel electrophoresis（DGGE） bands and richness index（S） were significantly affected by the interaction between F：C and FL（P〈0.05）, under high F：C, band numbers and richness index from WL group were higher than that from SL group（P〈0.05）, whereas there were no differences between WL andSL groups under low F：C（P〉0.05）. Microflora similarity was 50–73% among the different treatments. It is concluded that the WL with high F：C（65：35） diet is suitable for weaned calves.

Long-term effect of subacute ruminal acidosis on the morphology and function of rumen epithelial barrier in lactating goats

Grain-induced subacute ruminal acidosis(SARA)impairs rumen epithelial barrier function,but it is yet to be determined if SARA can cause persistent damage to the morphology and function of the rumen epithelial barrier.The objective of the present study was to investigate if SARA has persistent effects on the morphological structure and permeability of ruminal epithelium and the expression of the genes involved in epithelial barrier function using a lactating goat model.Twelve mid-lactating Saanen goats with rumen cannulas were randomly assigned to 1 of 2 groups:control group(Ctrl,n=4)fed a basal diet with a non-fiber carbohydrate(NFC)to neutral detergent fiber(NDF)ratio of 1.40,and SARA group(SARA,n=8)fed the same basal diet but with increasing NFC to NDF ratio from 1.4 to 1.79,2.31,and 3.23 overtime to induce SARA.At the end of the SARA challenge(post-SARA),4 goats were randomly selected from the SARA group and fed only hay mixture ad libitum for another 4 weeks to allow for restitution(post-SARA).Ruminal pH was continuously recorded to monitor the severity of SARA.Samples of the ventral ruminal epithelium were collected after slaughter to examine the structural and functional changes of the ruminal epithelium using transmission electron microscopy(TEM),Ussing chambers,qRT-PCR,and Western bolt analyses.Compared with the Ctrl group,ruminal papilla length,width,surface area and thickness of stratum corneum increased(P<0.05),while stratum spinosum and basale thickness,and total depth of the epithelium decreased(P<0.05)in the SARA group.These changes diminished or tended to return to the levels of the Ctrl group in the post-SARA group(P>0.05).The SARA challenge also decreased cellular junction and widened the intercellular space between epithelial cells.Rumen transepithelial short-circuit current(Isc),tissue conductance(Gt),and mucosa-to-serosa flux of paracellular horseradish peroxidase(HRP)all increased(P<0.05)both in the SARA and post-SARA groups,which indicates that SARA can induce a sustained increase in epithelial permeability and barrier dysfunction.Moreover,the mRNA and protein expressions of CLDN1,OCLN and ZO-1 were down-regulated(P<0.01)in both the SARA and post-SARA groups.The results of this study showed that SARA could result in sustained epithelial barrier dysfunction,at both structural and functional levels,which is associated with decreased expression of rumen epithelial tight junction proteins,and the restitution of rumen epithelial barrier function is slower than that of its morphology.

Effect of Soybean Peptides on in vitro Ruminal Fermentation and Microbial Population

This study was conducted to investigate the effect of Soybean Peptides(SPs) on ruminal fermentation and microbial diversity in vitro. Four levels of SPs supplements(0, 0.25%, 0.50% and 0.75% dry matter basis) were tested. p H, NH3-N, Neutral Detergent Fiber(NDF), Acid Detergent Fiber(ADF), Dry Matter Digestibility(DMD) and the Total Volatile Fatty Acid(TVFA) were measured at 6, 24, 48 h of in vitro mixed incubation the fluids. Microbial populations were determined at 24 h and Microbial Proteins(MCP) were determined at 24 and 48 h. The gas production after 48 h in vitro incubation presented linear growth(p<0.05) and the highest content was the level of 0.5% SPs(dry matter basis). NH3-N concentration reached the highest concentration with 0.75%(dry matter basis) at 48 h. p H linearly increased(linear, p<0.05) from 6 to 48 h. The digestion of DMD increased with increasing doses of SPs at 24 h. NDF and ADF linearly(p<0.01) decreased by adding SPs. The concentration of TVFA linearly increased(p<0.05) at 6, 24 and 48 h, and reached the highest concentration at the level of 0.75%(dry matter basis). SPs decreased(p<0.05) the molar proportion of acetate and propionate, respectively at 24 and 48 h. Acetat to Propionate ratio(A/P; linear, p<0.05) increased at 48 h, and reached the greatest value at the level of 0.75%(dry matter basis) at 48 h. The populations of rumen Ruminococcus flavefaciens(R. flavefaciens), Ruminococcusalbus(R. albus), Fibrobacter succinogenes(F. succinogenes), Butyrivibrio fibrisolvens(B. fibrisolvens), Streptococcus bovis(S.bovis), Ruminobacter amylophilus(R. amylophilus) and Succinimonas amylolytica(S. amylolytica) were analyzed based on the total rumen bacterial 16 S ribosomal deoxyribonucleic acid(r DNA). The relative abundance of R. flavefaciens and R. albus increased at 24 h, but the relative abundance of F. succinogenes decreased at this time. The incubation of SPs had no effect on the abundance of S. bovis and R. amylophilus. The relative abundance of B. fibrisolvens and S. amylolytica reached the greatest value(p<0.05) at middle doses of SPs inclusion at 24 h. The value of MCPs linearly increased(p<0.05) at 24 and 48 h. These results showed that SPs could improve in vitro fermentation and nutrient digestion of feed substrates, and had the ability to modulate the ruminal fermentation pattern by regulating the composition of functional rumen microbes. Hence, SPs might be a potential feed additive applied in the diets of ruminants.

Effects of Supplementation with Increasing Levels of Energy Concentrate on the Productive Response and Ruminal Digestion of Dairy Cows Grazing Lucerne Pasture

The aim of the study was to determine the effect of three levels of energy concentrate intake on dry matter (DM) and energy intake, milk yield and composition, rumen environment and pasture neutral detergent fiber (NDF) digestion. Twelve Holstein multiparous cows in early lactation (69.0 ± 5 days postpartum) producing 32.8 (±4.0) kg milk were assigned to three treatments at (kg/cow day) 3.5 (T3.5), 7.0 (T7.0) and 10.5 (T10.5) kg concentrate in a 3 × 3 Latin Square design. Parameters of ruminal environment and neutral detergent fiber (NDF) digestion were obtained using 3 additional rumen cannulated cows. Concentrate was composed (as fed) by corn grain (68%), soybean meal (22%), wheat bran (8%) and a vitamin-mineral premix including monensin and thoroughly consumed. Yields (kg/cow day-1) of milk, 4% fat corrected milk (4% FCM 4%) and energy corrected milk (ECM) resulted higher (p p -1 without effects on yield of milk fat. Concentrations (g/100 g) of milk fat (3.19), protein (2.97), total solids (11.75), non-fat solids (8.60) and casein (2.40) did not differ. Milk lactose content (g/100 g) was linearly increased (p p p p p < 0.05) in T3.5 due to the increase in acetate and butyrate while the acetate: Propionate ratio remained unchanged. Pasture NDF digestion was affected as concentrate intake increased. To increase milk protein yield and reduce concentrations of N-NH3 in rumen and milk, feeding an energy concentrate at 41% of total DM intake resulted an effective tool.

Effect of Molasses Addition at Ensiling on Ruminal In Situ Dry Matter and Nutrient Degradation of Whole-Plant Soybean Silage Harvested at Different Phenological Stages

Whole-plant soybean ensiling has limitations and challenges that affect silage fermentation patterns and reduce ruminal nutrient degradation. Perhaps either the addition of molasses at ensiling or harvesting at different phenological stages has the potential to enhance whole-plant soybean silage (WSS) ruminal degradation. This experiment was a completely randomized design with a 3 × 2 factorial arrangement of treatments evaluating the effects of molasses (0 and 40 g/kg fresh forage) and phenological stage (R5: beginning seed, R6: full seed, and R7: beginning maturity) on ruminal in situ degradation. Molasses increased effective ruminal degradability (ERD) of dry matter (DM) regardless of the phenological stage and increased ERD of crude protein at R5 and R6. The addition of molasses at later phenological stages increased ERD of neutral detergent fiber (NDF). There was a molasses × phenological stage interaction effect with greater fractions A and B of NDF at R7 and R5, respectively, but lower undegraded NDF at R5 and R6 with molasses addition. Although the benefits of adding molasses were more pronounced in R5 and R6, adding molasses to whole-plant soybean at ensiling is recommended regardless of phenological stage. Increasing the proportion of digestible nutrients at harvesting either by harvesting at later phenological stages or the addition of molasses is a powerful strategy to manage ruminal degradation of DM and nutrients in WSS.

Dietary Coleus amboinicus Lour. decreases ruminal methanogenesis and biohydrogenation, and improves meat quality and fatty acid composition in longissimus thoracis muscle of lambs

Background:Methane production and fatty acids(FA)biohydrogenation in the rumen are two main constraints in ruminant production causing environmental burden and reducing food product quality.Rumen functions can be modulated by the biologically active compounds(BACs)of plant origins as shown in several studies e.g.reduction in methane emission,modulation of FA composition with positive impact on the ruminant products.Coleus amboinicus Lour.(CAL)contains high concentration of polyphenols that may potentially reduce methane production and modulate ruminal biohydrogenation of unsaturated FA.This study aimed to investigate the effect of BAC of Coleus amboinicus Lour.(CAL)fed to growing lambs on ruminal methane production,biohydrogenation of unsaturated FA and meat characteristics.In this study,the in vitro experiment aiming at determining the most effective CAL dose for in vivo experiments was followed by two in vivo experiments in rumen-cannulated rams and growing lambs.Experiment 1(RUSITEC)comprised of control and three experimental diets differing in CAL content(10%,15%,and 20%of the total diet).The two in vivo experiments were conducted on six growing,rumen-cannulated lambs(Exp.2)and 16 growing lambs(Exp.3).Animals were assigned into the control(CON)and experimental(20%of CAL)groups.Several parameters were examined in vitro(pH,ammonia and VFA concentrations,protozoa,methanogens and select bacteria populations)and in vivo(methane production,digestibility,ruminal microorganism populations,meat quality,fatty acids profiles in rumen fluid and meat,transcript expression of 5 genes in meat).Results:CAL lowered in vitro methane production by 51%.In the in vivo Exp.3,CAL decreased methane production by 20%compared with the CON group,which corresponded to reduction of total methanogen counts by up to 28%in all experiments,notably Methanobacteriales.In Exp.3,CAL increased or tended to increase populations of some rumen bacteria(Ruminococcus albus,Megasphaera elsdenii,Butyrivibrio proteoclasticus,and Butyrivibrio fibrisolvens).Dietary CAL suppressed the Holotricha population,but increased or tended to increase Entodiniomorpha population in vivo.An increase in the polyunsaturated fatty acid(PUFA)proportion in the rumen of lambs was noted in response to the CAL diet,which was mainly attributable to the increase in C18:3 cis-9 cis-12cis-15(LNA)proportion.CAL reduced the mRNA expression of four out of five genes investigated in meat(fatty acid synthase,stearoyl-CoA desaturase,lipoprotein lipase,and fatty acid desaturase 1).Conclusions:Summarizing,polyphenols of CAL origin(20%in diet)mitigated ruminal methane production by inhibiting the methanogen communities.CAL supplementation also improved ruminal environment by modulating ruminal bacteria involved in fermentation and biohydrogenation of FA.Besides,CAL elevated the LNA concentration,which improved meat quality through increased deposition of n-3 PUFA.Highlight·Coleus amboinicus Lour.(CAL)into sheep diet decreased CH4emission.·CAL did not reduce nutrient digestibility,but inhibited the methanogen community.·CAL increased ruminal propionate proportion and decreased acetate/propionate ratio.·CAL elevated n-3 fatty acid concentration in ruminal fluid and meat.·Supplementation of CAL improved some meat quality traits.

Ruminal bacteria lipopolysaccharides:an immunological and microbial outlook

Lipopolysaccharides(LPS)are outer membrane components of Gram-negative bacteria made of three regions:the O-antigen;the core oligosaccharide;and a glucosamine disaccharide linked to hydroxy fatty acids,which is named lipid A.The number phosphate groups,and hydroxy fatty acid chains is associated with the immunopotency and the immunomodulatory activity of LPS,where six-acyl chain lipid A with two phosphate groups is found in virulent strains and five-or four-acyl chain lipid A with one phosphate group are found in non-virulent bacteria strains.Ruminal bacteria are predominantly Gram-negative and their LPS have not been thoroughly investigated.In the rumen,LPS is comprised of mixed ruminal LPS.Drawing upon a body of theoretical and applied work,this paper aims to critically review the scientific literature regarding single-species and mixed ruminal bacteria LPS,highlighting the importance of ruminal LPS to the host.Lastly,future research directions are suggested in order to further our understanding of the roles of LPS in the rumen.Possible suggestions for further understanding ruminal LPS include(1)in silico evaluation of major bacteria contributing to ruminal LPS,(2)structural characterization of LPS from prominent ruminal bacteria species,such as ruminal selenomonads and Megasphaera elsdenii,and,(3)ruminal epithelial tissue immune response evaluation from single-species and mixed ruminal LPS.In conclusion,this review identifies numerous areas for future research,including setting the basis for future modeling and simulation of host microbiome interactions in ruminants.

Heat stress impacts the multi-domain ruminal microbiota and some of the functional features independent of its effect on feed intake in lactating dairy cows

Background:Heat stress(HS)affects the ruminal microbiota and decreases the lactation performance of dairy cows.Because HS decreases feed intake,the results of previous studies were confounded by the effect of HS on feed intake.This study examined the direct effect of HS on the ruminal microbiota using lactating Holstein cows that were pair-fed and housed in environmental chambers in a 2×2 crossover design.The cows were pair-fed the same amount of identical total mixed ration to eliminate the effect of feed or feed intake.The composition and structure of the microbiota of prokaryotes,fungi,and protozoa were analyzed using metataxonomics and compared between two thermal conditions:pair-fed thermoneutrality(PFTN,thermal humidity index:65.5)and HS(87.2 for daytime and 81.8 for nighttime).Results:The HS conditions altered the structure of the prokaryotic microbiota and the protozoal microbiota,but not the fungal microbiota.Heat stress significantly increased the relative abundance of Bacteroidetes(primarily Gram-negative bacteria)while decreasing that of Firmicutes(primarily Gram-positive bacteria)and the Firmicutes-toBacteroidetes ratio.Some genera were exclusively found in the heat-stressed cows and thermal control cows.Some co-occurrence and mutual exclusion between some genera were also found exclusively for each thermal condition.Heat stress did not significantly affect the overall functional features predicted using the 16S rRNA gene sequences and ITS1 sequences,but some enzyme-coding genes altered their relative abundance in response to HS.Conclusions:Overall,HS affected the prokaryotes,fungi,and protozoa of the ruminal microbiota in lactating Holstein cows to a different extent,but the effect on the structure of ruminal microbiota and functional profiles was limited when not confounded by the effect on feed intake.However,some genera and co-occurrence were exclusively found in the rumen of heat-stressed cows.These effects should be attributed to the direct effect of heat stress on the host metabolism,physiology,and behavior.Some of the“heat-stress resistant”microbes may be useful as potential probiotics for cows under heat stress.

Effects of Supplemental Carbohydrate Sources on Lactation Performance, Nitrogen Balance,and Ruminal Fermentation in Dairy Cows Fed Energy-Deficient Total Mixed Rations

The effects of supplemental carbohydrate （CHO） sources on milk yield and composition, nitrogen （N） balance,and ruminal fermentation were evaluated in lactating dairy cows fed energy-deficient total mixed rations （TMR） . Fifty-six lactating Holstein cows （36. 8 ± 3. 4 kg / d of milk yield; 152 ± 26 days in milk [DIM]） were randomly assigned to one of four diets： （1） basal TMR （control） ; （2） basal TMR ＋ 1. 25 kg / d sucrose （SUC） ; （3） basal TMR ＋ 2. 12 kg /d steam-flaked corn （SFC） ; （4） basal TMR ＋ 2. 23 kg / d wheat （WHE） . The trial lasted 70 days,including the first 10 days of adaptation. Simultaneously, a 4 × 4 Latin square trial was conducted with four ruminally cannulated cows of similar DMI （Dry Matter Intake） fed on the above four diets. Each testing period lasted 18 days： 15 days for adaptation and 3 days for sampling. Cows fed SUC-,SFC-,and WHE-supplemented diets produced 1. 6,2. 9,and 0. 8 kg / d,respectively,more milk than those on the basal TMR control diet,but the difference was not significant （P = 0. 160） . However,the production of energy-corrected milk of cows fed CHO-supplemented diets improved significantly （P = 0. 020） ,and the yield of 4% fatcorrected milk tended to be higher （P = 0. 063） than in control animals. Percentages of milk protein,lactoseand solids-not-fat （SNF） increased significantly in cows fed CHO-supplemented diets （P 〈 0. 05） ,and yields of milk protein and SNF were significantly higher （P 〈0. 05） ; yields of milk fat and lactose were somewhat higher （P 〈 0. 10） . Cows receiving supplementary CHO diets consumed more DM （Dry Matter） （P = 0. 023） ; however,there was no significant difference in N retention,digestibility,or utilization efficiency among the treatments （P 〉 0. 10） . The average ruminal pH was lowest in cows supplemented with SUC,while that in cows supplemented with SFC and WHE was only slightly reduced （P = 0. 025） . Ruminal NH 3 -N concentration was highest with no supplementary CHO （P = 0. 017） ,and changes in postfeeding time were similar across the diets. Total volatile fatty acid concentration was significantly lower in control animals than in those on SUCand WHE-supplemented diets （P = 0. 001） ; significant dietary responses were observed in the concentrations of acetate,butyrate,and branchedchain fatty acids and in the acetate-to-propionate ratio （P 〈0. 001） . These results indicate that supplementation with different CHO sources in energy-deficient diets substantially improved the balance of energy and N in the rumen,altered ruminal fermentation,and improved lactation performance in dairy cows.

Effect of Hydropqinone on Ruminal Urease in the Sheep and its Inhibition Kinetics in Vitro

Effect of hydropuinone (HQ) on rumen urease acivity was studied. Hydroquinone at concentration of 0.01 mg·L -1 ,1 mg·L -1 and 10 mg·L -1 inhibited urease of intact rumen microbes in vitro by 25%, 34%, 55% and 64% respectively. In the present of low concentration of βmercaptoethanol,rumen urease could be solubilized and partially purified. The Km for the enzyme was 2×10 -3 mol·L -1 with Vmax of 319.144 μmoles/mg/min.The kinetics of inhibition with partially purified rumen urease was investigated.The result showed that the inhibuitory effect was not eliminated by increasing urea concentration indicating a noncompetitive in nature with inhibition constant 1.2×10 -5 mol·L -1 .Hydropuinone at a concentration that produced 64% urease inhibition did not affect ruminal total dehydrogenase, proteolytic enzyme( P >0.05)but increased cellulase activity by 28%( P <0.05)in vitro.These results demonstrated that hydropuinone was a specific inhibutor of rumen urease and could delay urea hydrolysis effectively without negative effect.The inhibitor appeared to offer the potential to improve nitrogen utilization by ruminants fed diets containing urea.