Ruminal acidosis and its definition: A critical review.

Ruminal acidosis occurs as a continuum of disorders, stemming from ruminal dysbiosis and disorders of metabolism, of varying severity. The condition has a marked temporal dynamic expression resulting in cases expressing quite different rumen concentrations of VFA, lactic acid, ammonia, and rumen pH over time. Clinical ruminal acidosis is an important condition of cattle and subclinical ruminal acidosis (SRA) is very prevalent in many dairy populations with estimates between 10 to 26% of cows in early lactation. Estimates of the duration of a case suggest the lactational incidence of the condition may be as high as 500 cases per 100 cows in the first 100 d of lactation. Historical confusion about the etiology and pathogenesis of ruminal acidosis led to definitions that are not fit for purpose as acidic ruminal conditions solely characterized by ruminal pH determination at a single point fail to reflect the complexity of the condition. Use of a model, based on integrated ruminal measures including VFA, ammonia, lactic acid, and pH, for evaluating ruminal acidosis is fit for purpose, as indicated by meeting postulates for assessing metabolic disease, but requires a method to simplify application in the field. While it is likely that this model, that we have termed the Bramley Acidosis Model (BAM), will be refined, the critical value in the model is that it demonstrates that ruminal acidosis is much more than ruminal pH. Disease, milk yield and milk composition are more associated with the BAM than rumen pH alone. Two single VFA, propionate and valerate are sensitive and specific for SRA, especially when compared with rumen pH. Even with the use of such a model, astute evaluations of the condition whether in experimental or field circumstances will be aided by ancillary measures that can be used in parallel or in series to enhance diagnosis and interpretation. Sensing methods including rumination detection, behavior, milk analysis, and passive analysis of rumen function have the potential to improve the detection of SRA; however, these may advance more rapidly if SRA is defined more broadly than by ruminal pH alone.

Postbiotics from Saccharomyces cerevisiae fermentation stabilize rumen solids microbiota and promote microbial network interactions and diversity of hub taxa during grain-based subacute ruminal acidosis (SARA) challenges in lactating dairy cows.

Background: High-yielding dairy cows are commonly fed high-grain rations. However, this can cause subacute ruminal acidosis (SARA), a metabolic disorder in dairy cows that is usually accompanied by dysbiosis of the rumen microbiome. Postbiotics that contain functional metabolites provide a competitive niche for influential members of the rumen microbiome, may stabilize and promote their populations, and, therefore, may attenuate the adverse effects of SARA. Methods: This study used a total of 32 rumen-cannulated lactating dairy cows, which were randomly assigned into four treatments: no SCFP (control), 14 g/d Original XPC (SCFPa), 19 g/d NutriTek (SCFPb-1X), and 38 g/d NutriTek (SCFPb-2X) (Diamond V, Cedar Rapids, IA) from 4 weeks before until 12 weeks after parturition. Grain-based SARA challenges were conducted during week 5 (SARA1) and week 8 (SARA2) after parturition by replacing 20% dry matter of the base total mixed ration (TMR) with pellets containing 50% ground barley and 50% ground wheat. The DNA of rumen solids digesta was extracted and subjected to V3-V4 16S rRNA gene sequencing. The characteristics of rumen solids microbiota were compared between non-SARA (Pre-SARA1, week 4; Post-SARA1, week 7; and Post-SARA2, weeks 10 and 12) and SARA stages (SARA1/1, SARA1/2, SARA2/1, SARA2/2), as well as among treatments. Results: Both SARA challenges reduced the richness and diversity of the microbiota and the relative abundances of the phylum Fibrobacteres. Supplementation with SCFP promoted the growth of several fibrolytic bacteria, including Lachnospiraceae UCG-009, Treponema, unclassified Lachnospiraceae, and unclassified Ruminococcaceae during the SARA challenges. These challenges also reduced the positive interactions and the numbers of hub taxa in the microbiota. The SCFPb treatment increased positive interactions among microbial members of the solids digesta and the number of hub taxa during the SARA and non-SARA stages. The SCFPb-2X treatment prevented changes in the network characteristics, including the number of components, clustering coefficient, modularity, positive edge percentage, and edge density of the microbiota during SARA challenges. These challenges reduced predicted carbohydrate and nitrogen metabolism in microbiota, whereas SCFP supplementation attenuated those reductions. Conclusions: Supplementation with SCFP, especially the SCFPb-2X attenuated the adverse effects of grain-based SARA on the diversity and predicted functionality of rumen solids microbiota.

Inhibiting methanogenesis by targeting thermodynamics and enzymatic reactions in mixed cultures of rumen microbes in vitro.

Mitigation of enteric methane (CH4) emissions from ruminant livestock represents an opportunity to improve the sustainability, productivity, and profitability of beef and dairy production. Ruminal methanogenesis can be mitigated via two primary strategies: (1) alternative electron acceptors and (2) enzymatic inhibition of methanogenic pathways. The former utilizes the thermodynamic favorability of certain reactions such as nitrate/nitrite reduction to ammonia (NH3) while the latter targets specific enzymes using structural analogs of CH4 and methanogenic cofactors such as bromochloromethane (BCM). In this study, we investigated the effects of four additives and their combinations on CH4 production by rumen microbes in batch culture. Sodium nitrate (NaNO3), sodium sulfate (Na2SO4), and 3-nitro-1-propionate (3NPA) were included as thermodynamic inhibitors, whereas BCM was included as a enzymatic inhibitor. Individual additives were evaluated at three levels of inclusion in experiments 1 and 2. Highest level of each additive was used to determine the combined effect of NaNO3 + Na2SO4 (NS), NS + 3NPA (NSP), and NSP + BCM (NSPB) in experiments 3 and 4. Experimental diets were high, medium, and low forage diets (HF, MF, and LF, respectively) and consisted of alfalfa hay and a concentrate mix formulated to obtain the following forage to concentrate ratios: 70:30, 50:50, and 30:70, respectively. Diets with additives were placed in fermentation culture bottles and incubated in a water bath (39°C) for 6, 12, or 24h. Microbial DNA was extracted for 16S rRNA and ITS gene amplicon sequencing. In experiments 1 and 2, CH4 concentrations in control cultures decreased in the order of LF, MF, and HF diets, whereas in experiments 3 and 4, CH4 was highest in MF diet followed by HF and LF diets. Culture pH and NH3 in the control decreased in the order of HF, MF, to LF as expected. NaNO3 decreased (p < 0.001) CH4 and butyrate and increased acetate and propionate (p < 0.03 and 0.003, respectively). Cultures receiving NaNO3 had an enrichment of microorganisms capable of nitrate and nitrite reduction. 3NPA also decreased CH4 at 6h with no further decrease at 24 h (p < 0.001). BCM significantly inhibited methanogenesis regardless of inclusion levels as well as in the presence of the thermodynamic inhibitors (p < 0.001) while enriching succinate producers and assimilators as well as propionate producers (p adj < 0.05). However, individual inclusion of BCM decreased total short chain fatty acid (SCFA) concentrations (p < 0.002). Inhibition of methanogenesis with BCM individually and in combination with the other additives increased gaseous H2 concentrations (p < 0.001 individually and 0.028 in combination) while decreasing acetate to propionate ratio (p < 0.001). Only the cultures treated with BCM in combination with other additives significantly (padj < 0.05) decreased the abundance of Methanobrevibacter expressed as log fold change. Overall, the combination of thermodynamic and enzymatic inhibitors presented a promising effect on ruminal fermentation in-vitro, inhibiting methanogenesis while optimizing the other fermentation parameters such as pH, NH3, and SCFAs. Here, we provide a proof of concept that the combination of an electron acceptor and a methane analog may be exploited to improve microbial efficiency via methanogenesis inhibition.

Effect of replacing concentrates with cassava root-top silage on feed utilization, rumen fermentation, blood parameters and growth performance in beef cattle.

OBJECTIVE: This experiment aimed to evaluate the effects of replacing concentrates with cassava root-top silage (CARTOS) on feed intake, digestibility, rumen fermentation, blood parameters, and growth performance of beef cattle. METHODS: Twenty crossbred bulls with a body weight (BW) of 226±56 kg were randomly assigned to one of five treatments for 90 d in a randomized complete block design having four blocks based on BW. The concentrates were replaced by CARTOS at levels of 0%, 25%, 50%, 75%, and 100% dry matter (DM). Animals were fed dietary treatments at 1.8% BW, with rice straw offered ad libitum. RESULTS: The DM and crude protein (CP) intake were decreased (p<0.01, p = 0.04) when the diet's CARTOS level was increased. The digestibility of DM, OM, and CP were not different among treatments, while fiber digestibility was increased with the inclusion of CARTOS (p = 0.03). The addition of CARTOS to replace concentrates did not change ruminal pH or volatile fatty acid proportions except for acetic acid, which increased with the addition of CARTOS (p = 0.03). The ruminal ammonia-nitrogen (NH3-N) was decreased (p<0.01) with increasing levels of CARTOS. The blood glucose and blood urea nitrogen decreased (p = 0.01) with the addition of CARTOS at 100%, whereas total protein and hematological parameters did not change with increasing levels of CARTOS. The use of CARTOS to substitute concentrates at 75% and 100% decreased average daily gain (ADG) and gain to feed ratio (G:F) (p<0.01); therefore, the addition of CARTOS up to 50% maintained ADG and G:F in beef cattle. CONCLUSION: CARTOS can replace concentrates up to 50% in beef cattle diets without adversely affecting feed intake, nutrient digestibility, rumen fermentation characteristics, blood parameters, or growth performance of beef cattle.

Feed characteristics and potential effects on ruminal bacteria of ensiled sugar kelp and winged kelp for Holstein dairy cows.

Seaweed silage has potential as an alternative feed ingredient for dairy cows. This study aims to investigate seaweed's and seaweed silages nutrient digestibility as well as their impact on the ruminal bacterial composition. The cultivated S. latissima and A. esculenta were preserved by freezing at - 40 °C or ensiling (16 °C, 3 months) with four different treatments: no additives, 4 g formic acid/kg wet seaweed, lactic acid bacteria (LAB) inoculant, and LAB inoculant in prewilted biomass (ca. 300 g DM/kg wet biomass). The nutrient digestibility was estimated using standard feed evaluation methods. The bacterial composition in ruminal fluid after 48 h in vitro anaerobic incubation with seaweeds and common feedstuffs was analysed using 16S ribosomal RNA (rRNA) amplicon sequencing (V3-V4) and quantitative PCR (qPCR). The results suggest that S. latissima was more digestible than A. esculenta and that the preservation treatments had only a small effect on the nutrient digestibility and ruminal bacteria compositions. The rumen DM degradability of S. latissima was comparable to common perennial and corn forage; however, the total tract CP digestibility of S. latissima (460 g/kg CP) was lower than common forages (620 - 820 g/kg CP) and was not improved by ensiling. There was a lack of insoluble but rumen-degradable CP in A. esculenta, making it unsuitable as a nutrient ingredient for dairy cows. The ruminal bacterial composition changed depending on the seaweed species used as substrate: The dominant bacterial taxa when incubated with S. latissima belonged to the genus Prevotella (relative abundance: 79 - 93%), known for its ability to degrade polysaccharides in various ecosystems. Moreover, the fibrolytic bacteria including Fibrobacter succinogenes and Ruminococcus flavefaciens were > 2.5 Log2FoldChange higher when incubating with S. latissima than with A. esculenta. These bacterial taxa may play an important role in the in vitro organic matter digestibility, noted as 2 times higher in S. latissima compared to A. esculenta. The qPCR results indicated potential methane mitigation properties of the studied seaweed species, with significantly lower gene copies of Archaea 16S rRNA and methyl coenzyme-M reductase subunit A genes when the ruminal fluid was incubated with the seaweed substrates. Our study suggested that ensiled S. latissima biomass can be included in the diet of dairy cows as an alternative forage-like ingredient with the potential of methane mitigation.

INVITED REVIEW: Research on ruminal biohydrogenation: Achievements, gaps in knowledge, and future approaches from the perspective of dairy science.

Scientific knowledge about ruminal biohydrogenation (BH) has improved greatly since this metabolic process was empirically confirmed in 1951. For years, BH had mostly been perceived as a process to be avoided to increase the post-ruminal flow of UFA from the diet. Two milestones changed this perception and stimulated great interest in BH intermediates themselves: In 1987, the in vitro anticarcinogenic properties of CLA were described, and in 2000, the inhibition of milk fat synthesis by trans-10 cis-12 CLA was confirmed. Since then, numerous BH metabolites have been described in small and large ruminants, and the major deviation from the common BH pathway (i.e., the trans-10 shift) has been reasonably well established. However, there are some less well-characterized alterations, and the comprehensive description of new BH intermediates (e.g., using isotopic tracers) has not been coupled with research on their biological effects. In this regard, the low quality of some published fatty acid profiles may also be limiting the advance of knowledge in BH. Furthermore, although BH seems to no longer be considered a metabolic niche inhabited by a few bacterial species with a highly specific metabolic capability, researchers have failed to elucidate which specific microbial groups are involved in the process and the basis for alterations in BH pathways (i.e., changes in microbial populations or their activity). Unraveling both issues may be beneficial for the description of new microbial enzymes involved in ruminal lipid metabolism that have industrial interest. From the perspective of diary science, other knowledge gaps that require additional research in the coming years are evaluation of the relationship between BH and feed efficiency and enteric methane emissions, as well as improving our understanding of how alterations in BH are involved in milk fat depression. Addressing these issues will have relevant practical implications in dairy science.

Ruminal passage rate and digestibility of fiber from dairy cows consuming diets containing alfalfa and orchardgrass hays with different concentrations of undegradable neutral detergent fiber (uNDF).

This study aimed to evaluate the effect of feeding diets with different fractions of undegraded NDF (uNDF) and potentially degradable (pdNDF) on ruminal NDF degradation and passage kinetics of lactating dairy cows. Six rumen-cannulated (533 ± 43 kg BW and 122 ± 15 DIM) and 6 non-cannulated (558 ± 62 kg BW and 126 ± 16 DIM) primiparous Holstein dairy cows were randomly assigned to 1 of 2 experimental diets in a crossover design with 2 28-d periods. The experimental diets were formulated to include either alfalfa hay (ALFA) or orchardgrass hay (ORCH) in addition to corn silage. Rations were formulated to contain 30% NDF (DM basis), where the concentrate, corn silage, and each of the hays provided one third of the dietary NDF. The marker dilution technique was used to measure the passage rate utilizing a pulse dose of marked corn silage fiber. On d 17 and 24 of each period, ruminal contents were evacuated to determine ruminal pool size. Following the return of the ruminal contents containing the pulse dose of marked corn silage to the rumen, ruminal grab samples were collected at 0, 3, 6, 9, 12, 24, 36, 48, 60, and 72 h. Samples from each time point were separated into solids and liquid, and the solids were analyzed for NDF, uNDF, and marker concentration. Alfalfa hay had a higher concentration of CP (16.4 vs. 10.7%) and a lower concentration of NDF (38.0 vs. 63.2) than orchardgrass hay. Alfalfa hay had a greater concentration of uNDF than orchardgrass hay (36.5 vs. 32.8% uNDF; NDF basis). Cows consuming the ALFA diet had similar milk yield (39.1 kg/d) and similar milk fat and protein concentrations (3.72% fat and 3.24% protein, respectively) than cows consuming the ORCH diet. Cows consuming the ALFA diet consumed more DM (26.7 vs. 24.6 kg/d) and uNDF (2.7 vs. 2.3 kg/d), than cows consuming the ORCH diet. Cows consuming the ALFA diet digested more NDF and pdNDF than cows consuming the ORCH diet (3.3 vs. 2.8 kg/d). Even though cows consuming the ALFA diet had a smaller pool size of NDF than cows consuming the ORCH diet (5.4 vs. 6.7 kg), the pool size of uNDF did not differ between groups (2.4 kg). Cows consuming the ALFA diet had a faster rate of passage of uNDF than cows consuming the ORCH diet (5.02 vs. 4.03%/h). This translated into a shorter mean retention time of uNDF for cows consuming the ALFA diet relative to cows consuming the ORCH diet (21.0 vs. 26.2 h). In conclusion, cows consuming diets containing alfalfa hay had a faster ruminal passage rate and a shorter mean retention time of uNDF than cows consuming diets containing orchardgrass hay, and this occurred despite the greater concentrations of dietary uNDF in the alfalfa-based diet. These findings suggest that the kinetics of ruminal digestion and passage influence NDF degradation in ways beyond uNDF concentration or forage quality.

Deciphering functional groups of rumen microbiome and their underlying potentially causal relationships in shaping host traits.

Over the years, microbiome research has achieved tremendous advancements driven by culture-independent meta-omics approaches. Despite extensive research, our understanding of the functional roles and causal effects of the microbiome on phenotypes remains limited. In this study, we focused on the rumen metaproteome, combining it with metatranscriptome and metabolome data to accurately identify the active functional distributions of rumen microorganisms and specific functional groups that influence feed efficiency. By integrating host genetics data, we established the potentially causal relationships between microbes-proteins/metabolites-phenotype, and identified specific patterns in which functional groups of rumen microorganisms influence host feed efficiency. We found a causal link between Selenomonas bovis and rumen carbohydrate metabolism, potentially mediated by bacterial chemotaxis and a two-component regulatory system, impacting feed utilization efficiency of dairy cows. Our study on the nutrient utilization functional groups in the rumen of high-feed-efficiency dairy cows, along with the identification of key microbiota functional proteins and their potentially causal relationships, will help move from correlation to causation in rumen microbiome research. This will ultimately enable precise regulation of the rumen microbiota for optimized ruminant production.

Moringa leaf meal exerts growth benefits in small ruminants through modulating the gastrointestinal microbiome.

This study investigated the impact of feeding 17% moringa leaf meal (MLM) on the ruminal and fecal microbial composition and body weight gain (BWG) performance of lambs (Ovis aries) and kids (Capra hircus). A total of n = 28 lambs (n = 14, no-moringa, n = 14, 17% moringa) and 24 kids (n = 12, no-moringa, n = 12, 17% moringa) were involved in the experiment and body weight was recorded fortnightly. Metagenomic shotgun sequencing was performed on 28, 22, and 26 ruminal solid, liquid fraction, and fecal samples from lambs, and 23, 22, and 23 samples from kids. Moringa supplementation significantly increased BWG in lambs (21.09 ± 0.78 to 26.12 ± 0.81 kg) and kids (14.60 ± 1.29 to 18.28 ± 1.09 kg) (p-value ≤ 0.01). Microbiome analysis revealed an elevated Firmicutes:Bacteroidetes ratio in the moringa diet group. Moringa-fed animals exhibited increased microbial genera associated with volatile fatty acids (VFAs) production (Prevotella, Anaerovibrio, Lachnospiraceae, Butyrivibrio, Christensenella) and starch and fiber digesters (Proteobacteria, Ruminococcus). The increase in the bacterial genus Sharpea suggested possible methane reduction and decreased proportion of pathogens, Aliarcobacter_ID28198, Campylobacter_ID194 and Campylobacter_ID1660076 suggest health benefits. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated significant alterations in microbial gene pool and metabolic pathways related to carbohydrate, protein, lipid and energy metabolism, indicating potential improvements in animal health. Overall, moringa feeding showed higher energy recovery, improved growth, and potential benefits in methane reduction and reduced pathogenic bacteria.

Comparative effects of nisin and monensin supplementation on growth performance, rumen fermentation, nutrient digestion, and plasma metabolites of fattening Hu sheep.

Introduction: This study was conducted to compare the effects of nisin (NIS) and ionophore antibiotic monensin (MON) on the growth performance, rumen fermentation, nutrient digestion and plasma metabolites of fattening Hu sheep. Methods: Thirty-six male Hu sheep (23.5 ± 1.0 kg) were divided into two blocks based on BW (low BW and high BW). Sheep within each block were then allotted to 9 pens respectively (two sheep/pen). Pens within each block were randomly assigned to one of three dietary treatments: (1) basal diet (CON); (2) basal diet + 40 mg/kg DM of MON; (3) basal diet + 274.5 mg/kg DM of NIS. The study lasted 9 weeks, with the initial 2 weeks for adaptation and the subsequent 7 weeks for treatment. Results: The results showed that both NIS and MON addition had no impacts on average daily gain (ADG), dry matter intake (DMI), and feed conservation rate (G:F) of sheep (p > 0.05). The digestibility of ether extract (EE) was lower in the MON-fed and NIS-fed sheep (p < 0.01) than in the CON group, whereas crude protein (CP) digestibility was higher in the MON-fed sheep compared to those fed NIS (p < 0.05). Both NIS and MON supplementation decreased acetate levels and acetate/propionate ratio in the rumen of Hu sheep (p < 0.05). Sheep fed MON exhibited higher total cholesterol concentrations (p < 0.05) compared to the CON and NIS groups. However, there were no significant differences in other plasma metabolites, including blood urea nitrogen (BUN), total bile acid, triglyceride, total protein, albumin, globulin, glucose, etc., among the three groups (p > 0.05). Discussion: In conclusion, dietary addition of NIS and MON altered the rumen fermentation mode by reducing acetate levels, with no discernible effects on the growth performance of the fattening Hu sheep.

Effects of rumen-protected 5-hydroxytryptophan on circulating serotonin concentration, behaviour, and mammary gland involution in goats.

The risk of acquiring new intramammary infections is high at the end of lactation, especially for the high milk-producing dairy animals. Resistance to bacterial infection increases following the completion of mammary gland involution after milking cessation. The serotonin precursor 5-hydroxytryptophan (5-HTP) could accelerate involution by increasing circulating serotonin levels, but ruminal microbes may degrade 5-HTP if orally administered to adult ruminants. It is unclear whether rumen-protected 5-HTP could effectively mediate circulating serotonin (5-hydroxytryptamine, 5-HT) and therefore accelerate mammary gland involution in ruminants. Goats were used as a model in the current study to investigate the effects of rumen-protected 5-HTP on behaviour, 5-HT metabolism, and mammary involution in ruminants. In the first experiment, 16 female Dazu black goats were assigned to one of four groups in a randomised block design. The treatments included a basal diet plus 0, 4, 20, or 100 mg/kg BW of rumen-protected 5-HTP. Serum was collected at 0, 3, 6, 12, and 24 h after offering the rumen-protected 5-HTP in the morning feed, and the behaviours were monitored. In the second experiment, 12 female Dazu black goats (Somatic cell count < 250 000) were randomly assigned to the control (basal diet) or rumen-protected 5-HTP group (basal diet plus 20 mg/kg BW). Milk or mammary secretions were manually collected aseptically on d -1, 1, 2, 3, 4, and 5 around weaning. The results depicted that rumen-protected 5-HTP supplementation elevated circulating 5-HTP and 5-hydroxyindole acetic acid concentrations, while 20 mg/kg BW of rumen-protected 5-HTP supplementation lowered the goats' locomotive activity. A high concentration of rumen-protected 5-HTP (100 mg/kg BW) increased serum alkaline phosphatase and gamma-glutamyl transpeptidase concentrations. Moreover, oral supplementation with 20 mg/kg BW of rumen-protected 5-HTP accelerated mammary gland involution and reduced feed intake in goats after weaning. These results demonstrate that oral supplementation with rumen-protected 5-HTP influences 5-HT metabolism and accelerates mammary gland involution after milking cessation in ruminants.

Analysis of the relationship between energy balance and properties of rumen fermentation of primiparous dairy cows during the perinatal period.

Short-chain fatty acids (SCFAs) produced in the rumen are key factors affecting dairy cows' energy balance (EB). This study aimed to quantitatively evaluate the effects of SCFAs production on EB in dairy cows. Primiparous dairy cows were divided into high non-esterified fatty acid (NEFA; group H) and low NEFA (group L) groups based on their blood NEFA levels at week 3 postpartum, which served as an indicator of EB. The amounts of SCFAs produced in the rumen, including acetate, propionate, and butyrate (SCFAsP), were calculated using the predicted rumen volume. Because there were no differences between the groups in SCFAsP/dry matter intake, whereas 4% fat-corrected milk (FCM)/SCFAsP was significantly higher in group H, it was suggested that more body fat was mobilized for milk production in group H. However, group L, which showed better EB, had propionate dominant and lower FCM/SCFAsP and milk energy/SCFAs energy at 3 and 7 weeks postpartum, indicating that group L had a better energy supply for milk production. These results suggest that SCFAsP produced by rumen fermentation and the composition of SCFAs in the rumen affect milk production and EB.

Effect of the dietary supplementation with sunflower oil-enriched bromoform from Asparagopsis taxiformis on lambs' growth, health, and ruminal methane production.

The red seaweed Asparagopsis taxiformis has a potent antimethanogenic effect, which has been proven both in vitro and in vivo. Vegetable oil immersions of this seaweed (hereafter Bromoil) help stabilise the bromoform (CHBr3) responsible for its antimethanogenic effect. We evaluate the effects of increasing the levels of CHBr3 in lamb diets on growth performance, methane (CH4) production, animal health and meat quality. Twenty-four Merino Branco ram lambs were fed a ground complete compound feed, supplemented with 50 mL/kg DM of sunflower oil with different CHBr3 content. The treatments were defined by the CHBr3 doses in the oil: 0 mg (control - B0), 15 mg (B15), 30 mg (B30) and 45 mg (B45) of CHBr3 per kg of feed DM. The feed was prepared daily by mixing Bromoil with the compound feed. At the end of the experiment, the lambs were sacrificed, the ruminal content was collected for in vitro fermentation to evaluate CH4 production and organic matter (OM) degradability, and the rumen mucosa was sampled for histological examination. Meat samples were collected for chemical composition and CHBr3 analysis. The half-life of CHBr3 in the air-exposed feed was 3.98 h making it very difficult to establish the practiced level of CHBr3 supplementation. Lambs-fed treatments B30 and B45 decreased DM intake by up to 28%. Average daily gain was also reduced due to CHBr3 supplementation, with B45 showing results 40% lower than B0. DM feed conversion ratio was similar for all treatments. The degradability of OM, the volume of total gas and of gas without CH4 were unaffected by the experimental treatments, evaluated by the in vitro method. However, the volume of CH4 decreased by up to 75% for treatments above 30 mg/kg DM, while the yield of CH4/g OM degraded was reduced by up to 78% with treatments above 30 mg/kg DM. Meat chemical composition was not affected by Bromoil supplementation and no traces of CHBr3 were found in meat samples. During this experiment, the animals presented normal health and behaviour. However, postslaughter examination of the rumen showed distinct lesions on the ventral region of the rumen mucosa of animals supplemented with Bromoil. These lesions were more severe in the animals receiving treatments B30 and B45. This research determined that although concentrations of CHBr3 in the diet above 30 mg/kg DM helped to reduce CH4 emissions, it negatively affected the performance and rumen wall.

Adding rumen microorganisms to improve fermentation quality, enzymatic efficiency, and microbial communities of hybrid Pennisetum silage.

Hybrid Pennisetum, a top biomass energy source, faces usage limitations because of its scarce lactic acid bacteria and high fiber content. This study assessed the influence of rumen fluid pretreatment on hybrid Pennisetum's silage, with focus on silage duration and rumen fluid effects on quality and fiber decomposition. Advanced third-generation sequencing was used to track microbial diversity changes and revealed that rumen fluid considerably enhanced dry matter, crude protein, and water-soluble carbohydrates, thus improving fermentation quality to satisfactory pH levels (3.40-3.67). Ideal results, including the highest fiber breakdown and enzymatic efficiency (47.23 %), were obtained with 5 % rumen fluid in 60 days. The addition of rumen fluid changed the dominant species, including Paucilactobacillus vaccinostercus (0.00 % vs. 18.21 %) and Lactiplantibacillus plantarum (21.03 % vs. 47.02 %), and no Enterobacter was detected in the high-concentration treatments. Moreover, strong correlations were found between specific lactic acid bacteria and fermentation indicators, revealing the potential of achieving efficient and economically beneficial hybrid Pennisetum production.

Effects of fermented jujube powder on growth performance, rumen fermentation, and antioxidant properties of simmental bulls.

Introduction: Fermented jujube powder (FJP) promotes a balance between the intestinal microflora and immune factors in animals. In this study, we aimed to investigate the effects of FJP on the production performance, nutrient digestion, rumen fermentation, and antioxidant properties of bulls. Methods: Forty Simmental bulls were randomly divided into four groups based on body weight and fed a basal diet with [5, 7.5, or 10% dry matter (DM)] or without FJP. The experimental period was 20 d for adaptation and 60 d for the feeding trial. Results: Dietary FJP supplementation did not affect DM intake (P > 0.05) but increased the average daily gain quadratically (P = 0.049) and decreased the feed conversion ratio linearly (P = 0.042). FJP quadratically enhanced DM and crude protein digestibility (P = 0.026 and P = 0.041, respectively) and linearly enhanced acid detergent fiber digestibility (P = 0.048). It also increased the total volatile fatty acid concentration quadratically (P = 0.037), acetate molar percentage, and acetate-to-propionate ratio linearly (P = 0.002 and 0.001), and reduced the ammonia nitrogen concentration linearly (P = 0.003). Additionally, xylanase and protease activities and Ruminococcus flavefaciens abundance increased linearly (P = 0.006, 0.018, and 0.009, respectively), and total bacteria, Ruminococcus albus, and Ruminobacter amylophilus abundance increased quadratically (P = 0.047, 0.011, and 0.021, respectively). FJP linearly increased serum total protein concentration and antioxidant capacity (P = 0.003 and 0.018, respectively) and decreased malonaldehyde content (P = 0.006). Discussion: FJP supplementation (7.5%) enhanced production performance, nutrient digestion, rumen fermentation, and serum antioxidant capacity in bulls. The improved nutrient digestion may be due to an increase in ruminal microorganisms and total volatile fatty acids from the FJP. High blood antioxidant levels indicate that FJP may preserve proteins, thereby boosting the production performance of bulls.

High concentrate diets altered the structure and function of rumen microbiome in goats.

This study used metatranscriptomics to investigate the effects of concentrate diet level on rumen microbiome composition and function in goats. A total of 12 healthy 120-day-old Da'er goats were randomly allotted into two treatments: L group (low dietary concentrate level group, concentrate: forage ratio was 25: 75) and H group (high dietary concentrate level group, concentrate: forage ratio was 80: 20). The study included a 10-day pre-feeding period and a 60-day growth experiment. The results showed that compared with the L group, the average daily gain and the slaughter rate in the H group were increased, while the F/G was decreased; the concentration of lactate and ammonia nitrogen, and the proportion of butyrate and valerate in the rumen of the H group were increased, while the proportion of acetate, and the ratio of acetate to propionate were decreased (p < 0.05). Among rumen bacteria, compared with the L group, the H group significantly decreased the relative abundance of Firmicutes and Fibrobacteria at the phylum level, decreased the relative abundance of Bacteroidetes, Fibrobacter, and Sarcina and increased the relative abundance of Clostridium at the genus level, and decreased the relative abundance of Fibrobacter succinogenes, Sarcina sp. DSM 11001, Oscillibacter sp. KLE 1728, and Ruminococcus flavefaciens and increased the relative abundance of Clostridium sp. ND2 and Firmicutes bacteria CAG: 103 at the species level (p < 0.05). Among rumen fungi, the relative abundance of Basidiomycota, Neocallimastigomycota, Mortierella, Mortierella elongata, and Gonapodyna prolifera was lower in the H group than that in the L group (p < 0.05). Functional annotation results showed that the abundance of Glycoside hydrolases genes in rumen microbiome was significantly decreased in the H group compared to the L group (p < 0.05). The result of KEGG DEGs enrichment analysis showed that the gene expression of cellulose 1,4-ß-cellobiosidase, acetyl-CoA hydrolase, lactate dehydrogenase, succinate-semialdehyde dehydrogenase, D-malate dehydrogenase and related genes in methane production pathways of rumen microbiome was decreased in the H group. In summary, feeding high concentrate diets improved the production performance of goats, altered the structure and composition of rumen microbiome and changed the function of rumen microbiome.

Impact of ginger powder (Zingiber officinale) supplementation on the performance, biochemical parameters, antioxidant status, and rumen fermentation in Ossimi rams.

Background and Aim: Ginger (Zingiber officinale) has great potential as a growth promoter and immunostimulant in ruminant nutrition. This study assessed the impact of ginger powder supplementation on Ossimi rams' rumen fermentation, biochemical parameters, and antioxidant levels. Materials and Methods: Fifteen Ossimi rams, aged 10 ± 1.3 months and weighing 30 ± 1.5 kg. Rams were randomly divided into three experimental groups: The control group (G1) received standard feed, while ginger powder (5 g and 7 g/kg body weight [BW] for G2 and G3, respectively) mixed in water was administered to groups G2 and G3 before their standard feed. Results: The control group recorded higher dry matter (DM) intake values (p < 0.05) than the ginger-treated groups. The ginger-treated groups showed superiority (p < 0.05) in weight gain and feed conversion compared to the control group. The digestion coefficients of DM, crude protein, and crude fiber were significantly (p < 0.05) increased by a high dose (7 g/Kg BW) of ginger supplementation, whereas organic matter, ether extract, and nitrogen-free extract digestibility remained unchanged. Compared to the control group, the rams given 5 g of ginger had significantly less (p < 0.05) total protein and globulin in their serum, but the rams given 7 g of ginger had significantly more (p < 0.05) of these proteins. In the ginger groups, these levels were significantly (p < 0.01) lower than those in the control group for serum creatinine, uric acid, urea, total lipids, triglycerides, total cholesterol, glucose, serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase. Rams given ginger had significant growth hormone, insulin-like growth factor-1, total superoxide dismutase, GSH-Px, TAC, immunoglobulin (Ig) A, and IgG enhancement (p < 0.01), and a decrease (p < 0.01) in malondialdehyde concentration compared to the control group. Significant increases in total short-chain volatile fatty acids, acetic, propionic, and isovaleric acids (p < 0.05), and significant decreases in NH3N and protozoa (p < 0.01). Conclusion: Ginger powder (5 g and 7 g) can improve growth, immune responses, antioxidant status, and ruminal parameters in rams. Further study is needed to evaluate the effect of ginger on different types of animals (cow, buffalo, and goat) to develop new feed additives.

Effects of yeast culture supplementation on milk yield, rumen fermentation, metabolism, and bacterial composition in dairy goats.

The effects of yeast culture (YC) on dairy goat milk yield and potential effects of rumen microbial population changes on rumen fermentation are poorly understood. This study aimed to evaluate the effects of YC on milk yield and rumen fermentation in dairy goats and explore the potential microbial mechanisms. Forty Laoshan dairy goats with a weight of 51.23 ± 2.23 kg and daily milk yield of 1.41 ± 0.26 kg were randomly divided into 4 groups: control (no YC), YC1 (10 g/day per goat), YC2 (25 g/day per goat), and YC3 (40 g/day per goat). The pre-feeding period was 15 days, and the official period was 60 days. Laoshan dairy goats were milked twice daily, and the individual milk yield was recorded. On the last day of the official period, rumen fluid was collected to measure rumen fermentation, perform quantitative polymerase chain reaction (PCR), and detect metabolites. Compared to the control group, the YC group had greater milk yield; higher acetic acid, butyric acid, and total volatile fatty acid contents; and lower ammonia-N (NH3-N) content in the rumen (p < 0.05). YC increased the abundance of Clostridia_UCG-014 and Paraprevotella (p < 0.05). Differential metabolites L-leucine and aspartic acid were screened. This study revealed the microbial mechanisms linking the relative abundance of Paraprevotella and Clostridia_UCG-014 to L-leucine and aspartic acid utilization. These results describe the potential benefits of supplementing 10 g/day per goat YC in the diets of Laoshan dairy goats for improving the rumen environment and milk yield.

Structural, Biochemical, and Phylogenetic Analysis of Bacterial and Fungal Carbohydrate Esterase Family 15 Glucuronoyl Esterases in the Rumen.

Glucuronoyl esterases (GEs) are carbohydrate active enzymes in carbohydrate esterase family 15 which are involved in the hydrolysis of lignin-carbohydrate complexes. They are encoded by a wide range of aerobic and anaerobic fungi and bacteria inhabiting diverse environments. The rumen microbiome is a complex microbial community with a wide array of enzymes that specialize in deconstructing plant cell wall carbohydrates. Enzymes from the rumen tend to show low similarity to homologues found in other environments, making the rumen microbiome a promising source for the discovery of novel enzymes. Using a combination of phylogenetic and structural analysis, we investigated the structure-function relationship of GEs from the rumen bacteria Fibrobacter succinogenes and Ruminococcus flavefaciens, and from the rumen fungus, Piromyces rhizinflata. All adopt a canonical α/ß hydrolase fold and possess a structurally conserved Ser-His-Glu/Asp catalytic triad. Structural variations in the enzymes are localized to loops surrounding the active site. Analysis of the active site structures in these enzymes emphasized the importance of structural plasticity in GEs with non-canonical active site conformations. We hypothesize that interkingdom HGT events may have contributed to the diversity of GEs in the rumen, and this is demonstrated by the phylogenetic and structural similarity observed between rumen bacterial and fungal GEs. This study advances our understanding of the structure-function relationship in glucuronoyl esterases and illuminates the evolutionary dynamics that contribute to enzyme diversity in the rumen microbiome.

Effect of Different Slow-Release Urea on the Production Performance, Rumen Fermentation, and Blood Parameter of Angus Heifer.

This study investigated the effect of replacing part of the dietary soybean meal with either polymer-coated urea or gelatinized starch urea on the production performance, blood indexes, and ruminal fermentation of Angus heifers. A total of 210 purebred Angus cattle (BW = 314.26 kg) were divided into three groups: the no urea group (CON), the polymer-coated urea group (PCU), and the gelatinized starch urea group (GSU); 20 g/kg polymer-coated urea or 25 g/kg gelatinized starch urea was used to replace part of soybean meal in the concentrate feed, according to the principle of isometabolic energy and isonitrogenous. The result showed that the PCU group had higher ADG and ADF apparent digestibility, while it had a lower feed-weight ratio. On the 86th day, the serum albumin (ALB) content in the PCU group was significantly higher than that in the CON group. In rumen, compared with the CON group, the contents of acetic acid and total volatile fatty acid were significantly higher in the PCU group, whereas butyric acid and propionic acid were significantly higher in the PCU group and GSU group. Ruminal bacterial diversity analysis found that the abundance of Firmicutes was higher in the PCU group at the phylum level, and an inverse result was observed in Bacteroidetes. The abundance of Paraprevotella was higher in the PCU group, whereas higher abundance of Prevotella was found in the GSU group at the genus level. These results indicate that slow-release urea can replace part of soybean meal in the diet, and the amount of substitution in this trial had no diverse effect on the performance of Angus heifers.