Rumen Microbiota Research Articles

Dietary Additive Combination for Dairy Calves After Weaning Has a Modulating Effect on the Profile of Short-Chain Fatty Acids in the Rumen and Fecal Microbiota

Dietary Additive Combination for Dairy Calves After Weaning Has a Modulating Effect on the Profile of Short-Chain Fatty Acids in the Rumen and Fecal Microbiota

Substituting imported soybean meal with locally produced novel yeast protein in concentrates for Norwegian Red dairy cows: implications for rumen microbiota and fatty acid composition.

This research paper addresses the hypothesis that substituting soybean meal with locally produced yeast protein from Cyberlindnera jadinii in barley-based concentrates for Norwegian Red (NR) dairy cows does not have adverse effects on milk fatty acid (FA) composition, rumen microbiota and sensory quality of milk. As soybeans also represent valuable protein sources for human consumption, alternative protein sources need to be investigated for animal feed. A total of 48 NR dairy cows were allocated into three feeding treatments, with the same basal diet of grass silage, but different concentrates. The concentrates were all based on barley, but 7% of the barley in the barley-concentrate (BAR; negative control) was replaced by either soybean meal (SBM; conventional control) or yeast microbial protein (YEA). The experiment lasted for a total of 10 weeks, including 2 weeks of adaptation with the soybean meal concentrate. Analysis of the feed revealed some differences in the FA composition of the YEA concentrate compared to the SBM and BAR concentrates. In milk, only two FAs (C17:1n-8cis9 and an unidentified isomer of C18:3) were significantly different between the YEA- and SBM-group, while six FAs differed between the BAR- and SBM-group. However, the amount of these FAs was low compared to the entire FA profile (<0.7 g/100 g). The experimental diets did not affect rumen microbiota nor the milk sensory quality. This study shows that C. jadinii can replace soybean meal as a protein source in concentrates (7% inclusion) for NR dairy cows fed a diet composed of grass silage and concentrates without any effects on rumen microbiota, and without compromising the FA composition or sensory quality of milk.

Effect of different genetic backgrounds on rumen microbiota and serum metabolic phenotypes in beef cattle

Species with different genetic backgrounds exhibit distinct metabolic traits. Nine beef cattle were selected for the experiment to study changes in serum metabolic phenotypes, rumen microbiota diversity, and composition in beef cattle from different genetic backgrounds. Three groups were Chinese Simmental (S group), Simmental×Chinese Holstein (SH group), and Simmental × Mongolian (SM group) cattle. We used ELISA to detect serum biochemical indicators. The Short-chain fatty acids (SCFAs) in the rumen were examined, and a significant difference was observed in the acetic acid content of the three experimental groups (p < 0.01). The propionic acid content in the rumen of the S group was significantly higher than that of the SH and SM groups (p < 0.05). The A/P ratios of both the S and SM groups were significantly higher than that of the SH group (p < 0.05). We analyzed rumen microbiota composition and diversity in each group of cattle using 16 S rRNA sequencing and found that their composition was generally similar in the three groups of crossbred fattening cattle; however, the f_Bacteroidales_RF16_group and g_norank_f_Bacteroidales_RF16_group were significantly enriched in the SH group, whereas Treponema and Spirochaetia were significantly enriched in the SM group. Spirochaetia was significantly enriched in the SM group. Differences in rumen bacterial enrichment indicated that starch, protein, and cellulolytic abilities differed among the S, SH, and SM groups. The results of Spearman correlation analysis confirmed the correlation between rumen genera and serum biochemical indices. Overall, differences in rumen microflora play an important role in influencing the serum metabolic phenotype.

Environmental Driving of Adaptation Mechanism on Rumen Microorganisms of Sheep Based on Metagenomics and Metabolomics Data Analysis.

Natural or artificial selection causes animals to adapt to their environment. The adaptive changes generated by the rumen population and metabolism form the basis of ruminant evolution. In particular, the adaptive drive for environmental adaptation reflects the high-quality traits of sheep that have migrated from other places or have been distant from their origins for a long time. The Hu sheep is the most representative sheep breed in the humid and low-altitude environments (Tai Lake region) in East Asia and has been widely introduced into the arid and high-altitude environments (Tibetan Plateau and Hotan region), resulting in environmental adaptive changes in the Hu sheep. In this study, a joint analysis of the rumen microbial metagenome and metabolome was conducted on Hu sheep from different regions (area of origin and area of introduction) with the objective of investigating the quality traits of Hu sheep and identifying microorganisms that influence the adaptive drive of ruminants. The results demonstrated that the growth performance of Hu sheep was altered due to changes in rumen tissue and metabolism following their introduction to the arid area at relatively high altitude. Metagenomic and metabolomic analyses (five ramsper area) revealed that 3580 different microorganisms and 732 different metabolites were identified in the rumen fluid of arid sheep. Among these, the representative upregulated metabolites were 4,6-isocanedione, methanesulfonic acid and N2-succinyl-L-arginine, while the dominant microorganism was Prevotella ruminicola. The downregulated metabolites were identified as campesterol, teprenone and dihydroclavaminic acid, while the disadvantaged microorganisms were Dialister_succinatiphilus, Prevotella_sp._AGR2160, Prevotella_multisaccharivorax and Selenomonas_bovis. The results of the Pearson analysis indicated that the rumen microbiota and metabolite content of sheep were significantly altered and highly correlated following their relocation from a humid lowland to an arid upland. In particular, the observed changes in rumen microorganisms led to an acceleration of body metabolism, rendering sheep highly adaptable to environmental stress. Prevotella_ruminicola was identified as playing an important role in this process. These findings provide insights into the environmental adaptation mechanisms of sheep.

GnRH Immunocastration in Male Xizang Sheep: Impacts on Rumen Microbiome and Metabolite Profiles for Enhanced Health and Productivity.

Castration is a prevalent and indispensable practice in sheep husbandry, aiding in enhancing meat quality, mitigating aggressive behavior, and managing unwanted reproduction. Nevertheless, the conventional surgical castration procedure poses several challenges, including heightened stress and pain, detrimental impacts on animal welfare, and diminished economic efficacy in farming operations. Consequently, immunocastration methods, serving as substitutes for surgical castration, are progressively finding application in livestock. The rumen, an essential and distinctive digestive and absorptive organ in ruminants, has been associated with enhanced meat quality and productive performance following castration in previous research studies, albeit fewer investigations have explored the potential impacts of GnRH immunization on the rumen's internal milieu in sheep post-de-escalation. Hence, the present study delved into evaluating the impact of GnRH immunocastration on the rumen microbiome and metabolomics in male Xizang sheep. This was achieved through the establishment of a GnRH immunocastration animal model and the collection of rumen fluid for microbiological and comprehensive metabolomics investigations. The outcomes of this investigation unveiled that the impact of GnRH immunocastration on body weight gain was more pronounced during the achievement of the castration objective. In addition, the Firmicutes-to-Bacteroidota ratio in the immune male (IM) group exceeded that of the control group (EM), suggesting that GnRH immunodeficiency may enhance the digestion and absorption of feed in male Xizang sheep. At the taxonomic level, the elevated presence of Prevotella and Quinella bacteria in the IM group compared to the EM group indicated that castration influenced a segment of the rumen microbiota in male Xizang sheep, thereby bolstering the digestive and metabolic efficacy of the rumen concerning nutrient utilization, particularly in the breakdown and absorption of proteins, carbohydrates, and lipids, ultimately expediting the fattening process and weight gain in male Xizang sheep following castration. Moreover, analysis of ruminal fluid metabolomics revealed that GnRH immunization had notable impacts on certain metabolites in the ruminal fluid of male Xizang sheep, with metabolites like 5-hydroxyindole acetic acid and 3-hydroxyindole acetic acid showing significant downregulation in the IM group compared to the EM group, while niacin and tyramine exhibited significant upregulation. These findings indicate a profound influence of GnRH immunization on the maintenance of ruminal equilibrium and ruminal health (including the health of ruminal epithelial cells). This study validates that GnRH immunocastration not only achieves the objectives of castration but also enhances ruminal health in male Xizang sheep, thus laying a foundational theoretical basis for the application and dissemination of GnRH immunocastration technology.

Unique rumen micromorphology and microbiota-metabolite interactions: features and strategies for Tibetan sheep adaptation to the plateau.

The rumen microbiota-a symbiont to its host and consists of critical functional substances-plays a vital role in the animal body and represents a new perspective in the study of adaptive evolution in animals. This study used Slide Viewer slicing analysis system, gas chromatography, RT-qPCR and other technologies, as well as 16S and metabolomics determination methods, to measure and analyze the microstructure of rumen epithelium, rumen fermentation parameters, rumen transport genes, rumen microbiota and metabolites in Tibetan sheep and Hu sheep. The results indicate that the rumen nipple height and cuticle thickness of Tibetan sheep are significantly greater than those of Hu sheep (p < 0.01) and that the digestion and absorption of forage are greater. The levels of carbohydrate metabolism, lipid metabolism, and protein turnover were increased in Tibetan sheep, which enabled them to ferment efficiently, utilize forage, and absorb metabolic volatile fatty acids (VFAs). Tibetan sheep rumen metabolites are related to immune function and energy metabolism, which regulate rumen growth and development and gastrointestinal homeostasis. Thus, compared with Hu sheep, Tibetan sheep have more rumen papilla and cuticle corneum, and the synergistic effect of the microbiota and its metabolites is a characteristic and strategy for adapting to high-altitude environments.

Multi-omics insights into the energy compensation of rumen microbiota of grazing yaks in cold season.

The ability of yaks to adapt to the extreme environment of low temperatures and hypoxia at cold seasons on the Qinghai-Tibet Plateau (QTP) is related to the host genome; however, the convergent evolution of rumen microbiomes in host adaption is unknown. Here, we conducted a multi-omics study on the rumen fluid of grazing yaks from warm (July) and cold (December) seasons on the QTP to evaluate the convergent evolution of rumen microbiomes in the adaptation of grazing yaks to cold-seasons environments. The results showed that grazing yaks at cold seasons had higher fibrolytic enzyme activities and volatile fatty acids (VFAs) concentrations, and the relative abundance of Firmicutes and the ratio Firmicutes to Bacteroidetes was significantly higher than that of yaks at warm seasons. Macrogenomic analyses showed that genes involved in forming VFAs and arginine were significantly enriched in cold-season yaks. Transcriptome analyses of the rumen epithelium showed that 72 genes associated with VFAs absorption and transport were significantly upregulated in cold-season yaks. Metabolomic analyses showed that the levels of ornithine, related to efficient nitrogen utilization, were significantly upregulated in cold-season yaks. The synergistic role of rumen microbiomes in the adaptation of grazing yaks to extreme environments at cold seasons was revealed by multi-omics study.

Dietary thiamine supplementation modulates ruminal microbiota and partly restores lactation performance in lactating Hu ewes under heat-stress conditions

This study aimed to investigate the impact of dietary thiamine supplementation on milk production, rumen microbiota profile and inflammatory response of Hu ewes under heat-stress conditions. Twenty-seven multiparous pregnant and lactating Hu ewes of 71.6 ± 1.4 kg body weight, 11 ± 0.5 months gestation age, 31 ± 1 days in milk, 1.68 ± 0.15 L/d milk yield, and 2.14 ± 0.12 kg of daily dry matter intake (DMI), were used in the study. Following an acclimation period of 7 days, ewes were divided into 3 blocks of 9 each (one Hu ewe per pen), in a complete randomized design and assigned to one of three environmental temperature conditions for 15 days: thermoneutrality without supplementation (CON), heat-stressed without supplementation (HS) or heat-stressed with dietary thiamine supplementation (200 mg/kg of DMI; HST). The results indicated that the HS group experienced notable declines in milk protein, fat, and lactose content from day 10–15 (P < 0.05) compared to the CON group. In contrast, thiamine supplementation significantly elevated milk protein content from day 13–15 and raised milk fat and lactose content on days 14 and 15 (P < 0.05) compared to the HS group, thereby enhancing milk quality to a certain extent. Additionally, thiamine supplementation (HST group) significantly increased the circulating thiamine and Immunoglobulin A (IgA) concentrations accompanied by a significant decline in LPS-binding protein, interleukin-1 (IL-1), and serum amyloid A protein (SAA) levels compared to the HS group (P < 0.05). Rumen microbial composition analysis revealed that thiamine supplementation increased the abundance of Pyramidobacter, Butyrivibrio, Bacteroides, and Saccharofermentans. In conclusion, heat exposure during lactation in Hu ewes lowered milk production and quality, affecting rumen microbes. Dietary thiamine supplementation could improve rumen fermentation, mitigate heat stress-induced inflammation, and restore milk production in Hu ewes under heat stress conditions.

Effect of fermented total mixed rations on rumen microbial communities and serum metabolites in lambs

AbstractBackgroundDiet regulates rumen microbiota, which in turn affects animal health. The present study evaluated the response of rumen microbiota and the immune system of lambs to a fermented total mixed ration diet.MethodsA total of 30 lambs were assigned into two groups: a group fed an unfermented high‐fiber diet (total mixed ration [TMR]) and a group fed an fermented low‐fiber diet (fermented TMR [FTMR]).ResultsThe results showed that FTMR markedly (p &lt; 0.05) increased average daily gain and dry matter intake compared to TMR. The FTMR diet increased the relative abundance of Veillonellaceae_UCG‐001 and decreased the diversity of undesirable microbiota despite stable overall microbial community diversity. Serum metabolomic analysis combined with enrichment analysis showed that serum metabolites were affected by the FTMR and metabolic pathways, and the FTMR diet significantly (p &lt; 0.05) influenced amino acid metabolism of lambs. There was a decrease in inflammatory factors in the FTMR treatment, indicating that inflammatory factors followed the same trajectory as changes in microbial community structure and function.ConclusionsOverall, the FTMR diet reduced undesirable microbiota diversity, thereby regulating host amino acid metabolism and improving immune status.

Effect of Dietary Benzoic Acid Supplementation on Growth Performance, Rumen Fermentation, and Rumen Microbiota in Weaned Holstein Dairy Calves

Supplementation with benzoic acid (BA) in animal feed can reduce feeds’ acid-binding capacity, inhibit pathogenic bacterial growth, enhance nutrient digestion, and increase intestinal enzyme activities. This study aimed to investigate the effects of different doses of BA on the growth performance, rumen fermentation, and rumen microbiota of weaned Holstein dairy calves. Thirty-two Holstein calves at 60 days of age were randomly assigned into four groups (n = 8): a control group (fed with a basal diet without BA supplementation; CON group) and groups that were supplemented with 0.25% (LBA group), 0.50% (MBA group), and 0.75% (HBA group) BA to the basal diet (dry matter basis), respectively. The experiment lasted for 42 days, starting at 60 days of age and ending at 102 days of age, with weaning occurring at 67 days of age. Supplementation with BA linearly increased the average daily gain of the weaned dairy calves, which was significantly higher in the LBA, MBA, and HBA groups than that in the CON group. The average daily feed intake was quadratically increased with increasing BA supplementation, peaking in the MBA group. Supplementation with BA linearly decreased the feed-to-gain (F/G) ratio, but did not affect rumen fermentation parameters, except for the molar proportion of butyrate and iso-butyrate, which were linearly increased with the dose of BA supplementation. Compared with the CON group, the molar proportions of iso-butyrate in the LBA, MBA, and HBA groups and that of butyrate in the HBA group were significantly higher than those in the CON group. Supplementation with BA had no significant effect on the alpha and beta diversity of the rumen microbiota, but significantly increased the relative abundances of beneficial bacteria, such as Bifidobacterium, and reduced those of the harmful bacteria, such as unclassified_o__Gastranaerophilales and Oscillospiraceae_UCG-002, in the rumen. Functional prediction analysis using the MetaCyc database revealed significant variations in the pathways associated with glycolysis across groups, including the GLYCOLYSIS-TCA-GLYOX-BYPASS, GLYCOL-GLYOXDEG-PWY, and P105-PWY pathways. In conclusion, BA supplementation improved the composition and function of rumen microbiota, elevated the production of butyrate and iso-butyrate, and increased the growth performance of weaned Holstein dairy calves.

The effect of different forages on rumen microbiota and milk production performance in Holstein dairy cows

The effect of different forages on rumen microbiota and milk production performance in Holstein dairy cows

Hybridization promotes growth performance by altering rumen microbiota and metabolites in sheep.

Hybridization can substantially improve growth performance. This study used metagenomics and metabolome sequencing to examine whether the rumen microbiota and its metabolites contributed to this phenomenon. We selected 48 approximately 3 month-old male ♂Hu × ♀Hu (HH, n = 16), ♂Poll Dorset × ♀Hu (DH, n = 16), and ♂Southdown × ♀Hu (SH, n = 16) lambs having similar body weight. The sheep were fed individually under the same nutritional and management conditions for 95 days. After completion of the trial, seven sheep close to the average weight per group were slaughtered to collect rumen tissue and content samples to measure rumen epithelial parameters, fermentation patterns, microbiota, and metabolite profiles. The final body weight (FBW), average daily gain (ADG), and dry matter intake (DMI) values in the DH and SH groups were significantly higher and the feed-to-gain ratio (F/G) significantly lower than the value in the HH group; additionally, the papilla height in the DH group was higher than that in the HH group. Acetate, propionate, and total volatile fatty acid (VFA) concentrations in the DH group were higher than those in the HH group, whereas NH3-N concentration decreased in the DH and SH groups. Metagenomic analysis revealed that several Prevotella and Fibrobacter species were significantly more abundant in the DH group, contributing to an increased ability to degrade dietary cellulose and enrich their functions in enzymes involved in carbohydrate breakdown. Bacteroidaceae bacterium was higher in the SH group, indicating a greater ability to digest dietary fiber. Metabolomic analysis revealed that the concentrations of rumen metabolites (mainly lysophosphatidylethanolamines [LPEs]) were higher in the DH group, and microbiome-related metabolite analysis indicated that Treponema bryantii and Fibrobacter succinogenes were positively correlated with the LPEs. Moreover, we found methionine sulfoxide and N-methyl-4-aminobutyric acid were characteristic metabolites in the DH and SH groups, respectively, and are related to oxidative stress, indicating that the environmental adaptability of crossbred sheep needs to be further improved. These findings substantially deepen the general understanding of how hybridization promotes growth performance from the perspective of rumen microbiota, this is vital for the cultivation of new species and the formulation of precision nutrition strategies for sheep.

Nitrogen fertiliser application rate to temperate mid-season grass: Digestion, nitrogen balance and rumen microbiota in beef cattle, and rumen fermentation and methane production in vitro

Nitrogen (N) excretion by cattle, particularly urinary N, can have detrimental environmental impacts on air and water quality, and contributes to greenhouse gas emissions. The effects of increasing the application rate of inorganic N fertiliser from 15 (N15) to 80 (N80) kg/ha per cut to Lolium perenne dominant swards in summer, on intake, rumen fermentation, rumen microbial populations, apparent total-tract digestibility and N-balance in beef cattle, and in vitro fermentation and methane output, were studied. Sixteen suckler-bred Charolais steers, used in a randomised block design experiment, were offered fresh grass mechanically harvested 21-d after N fertiliser application. Similar grass was incubated in an eight-vessel in vitro RUSITEC system. Grass crude protein concentration was 50 g/kg dry matter (DM) higher for N80 compared to N15. There was no difference in grass DM intake between treatments. Rumen fermentation variables did not differ between treatments, except for the molar proportion of propionate, which was greater for N80 than N15. Gross microbial community structure in the rumen was not significantly altered by inorganic N fertiliser application rate. The relative abundance of individual genera Lachnospiraceae_NK3A320_group and Ruminococcaceae_NK4A214_group, and Acetitomaculum were significantly lower at the higher N application rate. Mean plasma urea concentration was greater for N80 compared to N15. In vivo digestibility of DM, organic matter, neutral detergent fibre and acid detergent fibre was unaffected by fertiliser N application rate. Nitrogen intake was 75 g/d greater, and urinary and faecal N excretion were 20 and 5 g/d greater, respectively, for N80 than N15. The quantity of N retained and N use efficiency was greater for N80 compared to N15. In vitro NH3 concentration was greater for N80 than N15, whereas other rumen fermentation variables, and in vitro methane and total gas output, did not differ between treatments. Reducing the inorganic N fertiliser rate applied to mid-season temperate grass reduced N excretion from beef cattle, which is environmentally beneficial, with no effect on in vitro methane production.

Dietary secoisolariciresinol diglucoside crude extract improves growth through modulating rumen bacterial community and epithelial development in lambs.

Flaxseed lignans, types of polyphenolic compounds, primarily consist of secoisolariciresinol diglucoside (SDG). Natural plant extracts are becoming increasingly important as feed for ruminant animals. An underutilized plant bioactive component, SDG shows promising benefits for young ruminant production. The objective of this study was to assess the impact of SDG on rumen fermentation using an in vitro rumen simulation technology. Additionally, we tested the effects of SDG (0.20 g kg-1 body weight) on rumen development and production performance of lambs in a production setting. The in vitro addition of 100 mg L-1 SDG demonstrated significant regulatory effects, with a notable decrease in the acetate/propionate ratio (P < 0.05). Feeding trials revealed that SDG significantly increased average daily feed intake and average daily weight gain (P < 0.05), and reduced the acetate/propionate ratio (P < 0.05). This led to a significant increase in the relative abundance of Eubacterium ruminantium (P = 0.038) and Butyrivibrio (P = 0.002). Furthermore, it promoted rumen development and upregulated the relative expression of mRNA of Cyclin E1 and CDK2 in rumen epithelial cells (P < 0.05). The SDG extract optimizes the composition of rumen microbiota and the development of rumen epithelial cells, promoting the growth of pre-weaning lambs. The SDG additive exhibits potential as a novel growth promoter for ruminant animals, offering a promising solution for sustainable livestock production. © 2024 Society of Chemical Industry.

PSIX-30 Interaction between rumen microbiome and host feed efficiency phenotype across contrasting breeds and dietary sources

Abstract Provision of feed accounts for a high proportion of the variable cost in beef production systems, and consequently is a major determinant of overall profitability. Thus, improving the conversion of feed to animal product by identifying and breeding cattle with improved feed efficiency potential provides a method through which feed input costs may be reduced, whilst also contributing to more environmentally sustainable beef production. The rumen microbiome dictates the feed degradation capacity and subsequent nutrient supply in ruminants and is thus potentially impacted by feed efficiency phenotype. However, the relationship between rumen microbiota and host feed efficiency phenotype is yet to be fully elucidated. This is further complicated when contrasting diets, cattle breeds as well as various stages of animal development are considered. The objective of this study was to undertake network analysis on rumen microbiome data generated from two contrasting steer breed types (Charolais and Holstein-Friesian), divergent within-breed for residual feed intake (RFI), and offered contrasting diets during different stages of development: i) high-concentrate during the growing phase; ii) grass silage during the growing phase; iii) zero-grazed grass during the growing phase, and iv) high-concentrate during the finishing phase. From approximately 10 mo of age, Charolais (n = 90) and Holstein-Friesian (n = 77) steers were individually fed each of the aforementioned diets for 70 d, following an adaptation period between each diet. At the end of each dietary phase, RFI was determined for every steer and rumen fluid was sampled using a transesophageal sampling device from the 10 most-efficient (Low-RFI) and the 10 least-efficient (High-RFI) steers within each breed. Rumen fluid samples were then subjected to 16S rRNA sequencing and a network-based systems biology analysis subsequently undertaken using PCIT incorporating the 16S sequencing data and individual RFI values across each breed and dietary phase. A total of 110 microbial taxa were connected (P &amp;lt; 0.05; r &amp;gt; 0.8) to the breed and diet contrasts examined; however, no single microbe was commonly connected to all contrasts. The Pyramidobacter genus was the most represented phylotype, displaying a positive connection with the Charolais steers during the grass silage diet, as well as a negative association with Charolais steers during the first high-concentrate diet and Holstein-Friesian steers during the zero-grazed grass diet. Similarly, of the 35 microbial taxa significantly connected to more than one breed/diet contrast, 12 displayed altered direction of association, indicating a differential relationship between the rumen microbiome and RFI phenotype depending on the prevailing dietary management. Acknowledgement: The animal model used in this study was funded by the Irish Department of Agriculture, Food and the Marine (RSF13/S/519). Kate Keogh received funding for this work from the Research Leaders 2025 program (co-funded by Teagasc and the European Union’s Horizon 2020, Marie Skłodowska-Curie grant agreement number 754380).

400 Effect of pH on in vitro ruminal metabolism of trans-10 18:1 and trans-11 18:1

Abstract Dietary C18 unsaturated fatty acids are extensively biohydrogenated by rumen microbiota into 18:0 and trans-18:1 isomers. Usually, the biohydrogenation (BH) pathways produce mostly 18:0 and trans-11 18:1 as the main intermediate. However, when fed low-forage diets that decrease the rumen pH, the main BH intermediate often is the trans-10 18:1 isomer. This change in BH, called the trans-10 shift, is associated with milk fat depression, but despite that, little is known about the triggers of the trans-10 shift and its ecological role in rumen microbiota. Microbiota stressed by low rumen pH may benefit from increased trans-18:1 availability favoring the production of the trans-18:1 isomer with a slower rate of BH. Thus, we hypothesize that at low rumen pH conditions, the BH of trans-10 18:1 will be slower than that of trans-11 18:1. We tested that with an in vitro experiment using batch incubation in Hungate tubes, replicated in 5 wk. In each run, 24 tubes were allocated at 6 treatments and 4 incubation times (0, 6, 24 and 48 h). The treatments resulted from the combination of 2 buffering solution pH (6.74 vs. 5.85) and 3 substrates: 1) Control (60 mg feed DM) 2) T10 (Control plus 1.2 mg of trans-10 18:1) and 3) T11 (Control plus 1.2 mg of trans-11 18:1). Volatile fatty acids (VFA) and long-chain fatty acids were analyzed by gas chromatography. Data were analyzed using the 2 × 3 factorial arrangement for each incubation time. Low pH resulted in less 30 to 37% DM disappearance than those with high pH at all incubation times (P &amp;lt; 0.05). The decrease in VFA with low pH was less expressive (≈17%) and only evident at 24 and 48 h incubation times. The low pH also decreased (P &amp;lt; 0.05) the BH of 18:1n-9 (less ≈47 to 53%), 18:2n-6 (less 30 to 34%) and 18:3n-3 (less 20 to 22%) at 24h and 48h incubation times, respectively. The disappearance of trans-11 18:1 was 14.6% at 6 h, 29.0% at 24 h and 32.8% at 48 h and was not affected by pH. The disappearance of trans-10 18:1 was dependent on pH. At 6 h and high pH the disappearance of both was similar, but at low pH the disappearance of trans-10 18:1 was much less (9.8%) than that of trans-11 18:1. However, at 48 h and low pH, the disappearance was similar between isomers and the disappearance of trans-10 18:1 was much greater (46.6%) than trans-11 18:1. In summary, the BH of trans-10 18:1 is less at low pH than at high pH, and this could in part explain its abundance in some practical ruminant feeding conditions. Acknowledgements: Fundação para a Ciência e a Tecnologia (FCT) funding through PTDC/CAL-ZOO/29654/2017(RumOmics), PTDC/CAL-ZOO/4515/2021(Gene2Rumen), UIDB/00276/2020 (CIISA), and LA/P/0059/2020 (AL4Animals) projects.

93 Ruminal archaea and bacteria metatranscriptomic responses to supplementation in steers fed low-quality forage

Abstract Forage comprises the majority of beef cattle diets globally, and its quality can vary. Supplementation presents a significant opportunity to improve the efficiency of production. Supplementation has been shown to increase ruminally available nitrogen, improve forage utilization, and reduce livestock methane (CH4) emissions. However, corresponding effects of supplementation on gene expression of rumen microbiota, in particular metabolic pathways involved in methanogenesis, have yet to be studied. This study aimed to evaluate the effect of supplement composition before and after feeding on rumen archaeal and bacterial gene expression in steers consuming a low-quality forage diet. Ruminal samples were collected from five ruminally cannulated Angus steers [body weight (BW) = 375 ± 45 kg] used in a 4 × 4 Latin square fed a basal diet of King Ranch Bluestem Hay (3.5% CP, 73% NDF), and one of four isonitrogenous (130 mg N/kg BW) supplements. Steers provided the two most divergent supplements (2% starch, 43% CP; 56% starch, 21% CP) were used for metatranscriptomic analysis. Rumen fluid samples were collected via rumen cannula 0 and 4 h post-feeding. Total RNA was extracted and sequenced. Differentially expressed genes (DEGs) were identified and functionally annotated. Pathway enrichment analysis was completed to determine enriched pathways (broad metabolic pathways) and modules (specific metabolic reactions). Only main effects are presented in this abstract. Archaeal hydrogenotrophic and methylotrophic methanogenesis reactions and tryptophan biosynthesis reactions were upregulated in the 2% starch supplement (P &amp;lt; 0.05). The 2% starch supplement resulted in fewer bacterial total upregulated carbohydrate-active enzymes compared with the 56% starch supplement (957 vs. 1,695 DEGs, respectively; P &amp;lt; 0.05), and an increased proportion of cellulose and hemicellulose-specific enzymes (35 vs. 29%, respectively; P &amp;lt; 0.05). The 56% starch supplement resulted in upregulation of the bacterial 3-Hydroxypropionate bi-cycle reaction (P &amp;lt; 0.05). Collection time exerted a larger effect than supplement composition on both archaeal and bacterial gene expression, with 83% and 63% of DEGs associated with time, respectively (P &amp;lt; 0.05). Archaeal hydrogenotrophic methanogenesis and carbon metabolism were upregulated 4 h post-feeding. In addition, 1,704 bacterial carbohydrate-active enzymes were upregulated 4 h post-feeding, as compared with 975 upregulated at 0 h (P &amp;lt; 0.05). However, these differences in DEGs did not correlate with significantly enriched pathways of bacterial carbohydrate metabolism. These findings demonstrate improvements to low-quality forage utilization, through CP and starch supplementation, can be correlated to relevant changes in gene expression of rumen microbiota. Furthermore, they demonstrate that supplementation influences the expression of methanogenesis reactions, suggesting an opportunity to affect CH4 production. Additional studies quantifying forage utilization, rumen microbiota gene expression and CH4 production are needed to further optimize supplementation strategies for improved efficiency of beef production whilst maintaining or reducing its carbon footprint.

Succession of rumen microbiota and metabolites across different reproductive periods in different sheep breeds and their impact on the growth and development of offspring lambs

BackgroundThe microbiota and metabolites in the gastrointestinal tracts of female animals at different reproductive periods are very important to the growth, development, and health of themselves and their offspring. However, the changes in the gastrointestinal microbiota and metabolites throughout reproductive period of different sheep breeds and their effects on the growth and development of offspring lambs are still unclear. Hence, this study presents an assessment of the reproductive hormone levels, immune levels, rumen microbiota, and metabolites in Hu sheep and Suffolk ewes at different reproductive periods and their effects on the growth and development of offspring lambs.ResultsHu sheep and Suffolk during non-pregnancy, pregnancy, and lactation were used as the research objects to determine reproductive and immune indexes of ewes at different periods, analyze rumen microbiome and metabolome, and track the growth performance and development of offspring lambs. The results showed that the reproductive hormone and immune levels of Hu sheep and Suffolk underwent adaptive changes across different reproductive periods. Compared with non-pregnancy, the microbial energy metabolism and lipid metabolism function decreased during Hu sheep pregnancy, and energy metabolism function decreased during lactation. In Suffolk, energy metabolism, glycan biosynthesis, and metabolism function were enhanced during pregnancy, and the metabolism of cofactors and vitamins was enhanced during lactation. Prevotella increased in Suffolk during pregnancy and lactation (P < 0.05) and was positively correlated with the birth weight and body size of the lambs (P < 0.05). Moreover, the abundances of Butyrivibrio and Rikenellaceae_RC9_gut_group during pregnancy were positively correlated with the intestinal immunity of the offspring lambs (P < 0.05), thereby regulating the intestinal immunity level of the lambs. Metabolomic analysis revealed that the protein digestion, absorption, and amino acid metabolism of Hu sheep were enhanced during pregnancy, which provided amino acids for the growth and development of pregnant ewes and fetuses and was significantly correlated with the birth weight, body size, and intestinal immunity of lambs (P < 0.05). Simultaneously, there was an increase in acetate and propionate during the pregnancy and lactation period of both Hu sheep and Suffolk, providing energy for ewes during reproductive period. Moreover, the microbiota during the lactation period was significantly correlated with the milk quality and lambs daily gain (P < 0.05).ConclusionsThis study revealed the characteristic succession changes in the rumen microbiota and its metabolites at different reproductive periods in sheep breeds and their regulation of reproductive hormone and immune levels and identified their potential effects on the growth and development of offspring lambs. The findings provide valuable insights into the health and feeding management of different sheep breeds during the reproductive stage.Cnu7uEJBESQuA-vs1kGzfaVideo

Multi-omics analysis reveals the effects of host-rumen microbiota interactions on growth performance in a goat model.

The growth rate of young ruminants has been associated with production performance in later life, with recent studies highlighting the importance of rumen microbes in supporting the health and growth of ruminants. However, the specific role of rumen epithelium bacteria and microbiota-host interactions in influencing the early life growth rate of ruminants remains poorly understood. In this study, we investigated the rumen fermentation pattern, microbiota characteristics, and global gene expression profiles of the rumen epithelium in 6-month-old goats with varying growth rates. Our results showed that goats with high average daily gain (HADG) exhibited higher rumen propionate concentrations. Goats with low average daily gain (LADG) had the higher relative abundances of rumen epithelium bacteria genera U29-B03 and Quinella, while exhibiting a lower relative abundance of Lachnospiraceae UCG-009. In the rumen fluid, the relative abundances of bacteria genus Alloprevotella were lower and Desulfovibrio were higher in LADG goats compared to HADG goats. Additionally, the relative abundance of fungal genus Symmetrospora was lower in LADG goats compared to HADG goats. Transcriptome analysis showed that 415 genes were differentially expressed between LADG and HADG goats, which were enriched in functions related to cell junction and cell adhesion, etc. Correlation analysis revealed that rumen epithelium bacteria genera UCG-005 and Candidatus Saccharimonas were negatively associated, while Lachnospiraceae NK3A20 group and Oscillospiraceae NK4A214 group were positively associated with average daily gain (ADG) and genes related to barrier function. The rumen fluid bacteria genus Alloprevotella was positively correlated, while Desulfovibrio was negatively correlated with rumen propionate and ammoniacal nitrogen (NH3-N) concentrations, as well as genes related to barrier function and short chain fatty acids (SCFAs) transport. In summary, our study reveals that the higher ruminal fermentation efficiency, improved rumen epithelial barrier functions, and enhanced SCFAs transport in HADG goats could be attributed to the rumen microbiota, particularly the rumen epithelium bacteria, such as Lachnospiraceae and Oscillospiraceae NK4A214 group.

In vitro and ex vivo metabolism of chemically diverse fructans by bovine rumen Bifidobacterium and Lactobacillus species

BackgroundInulin and inulin-derived fructooligosaccharides (FOS) are well-known prebiotics for use in companion animals and livestock. The mechanisms by which FOS contribute to health has not been fully established. Further, the fine chemistry of fructan structures from diverse sources, such as graminan-type fructans found in cereal crops, has not been fully elucidated. New methods to study fructan structure and microbial responses to these complex carbohydrates will be key for evaluating the prebiotic potency of cereal fructans found in cattle feeds. As the rumen microbiome composition is closely associated with their metabolic traits, such as feed utilization and waste production, prebiotics and probiotics represent promising additives to shift the microbial community toward a more productive state.ResultsWithin this study, inulin, levan, and graminan-type fructans from winter wheat, spring wheat, and barley were used to assess the capacity of rumen-derived Bifidobacterium boum, Bifidobacterium merycicum, and Lactobacillus vitulinus to metabolize diverse fructans. Graminan-type fructans were purified and structurally characterized from the stems and kernels of each plant. All three bacterial species grew on FOS, inulin, and cereal crop fructans in pure cultures. L. vitulinus was the only species that could metabolize levan, albeit its growth was delayed. Fluorescently labelled polysaccharides (FLAPS) were used to demonstrate interactions with Gram-positive bacteria and confirm fructan metabolism at the single-cell level; these results were in agreement with the individual growth profiles of each species. The prebiotic potential of inulin was further investigated within naïve rumen microbial communities, where increased relative abundance of Bifidobacterium and Lactobacillus species occurred in a dose-dependent and temporal-related manner. This was supported by in situ analysis of rumen microbiota from cattle fed inulin. FLAPS probe derived from inulin and fluorescent in situ hybridization using taxon-specific probes confirmed that inulin interacts with Bifidobacteria and Lactobacilli at the single-cell level.ConclusionThis research revealed that rumen-derived Bifidobacteria and Lactobacilli vary in their metabolism of structurally diverse fructans, and that inulin has limited prebiotic potential in the rumen. This knowledge establishes new methods for evaluating the prebiotic potential of fructans from diverse plant sources as prebiotic candidates for use in ruminants and other animals.