Feeding live yeast (Saccharomyces cerevisiae) improved performance of mid-lactation dairy cows by altering ruminal bacterial communities and functions of serum antioxidation and immune responses.

BACKGROUND: The utilization of live yeast (Saccharomyces cerevisiae, YE) in dairy cows is gaining traction in dairy production as a potential strategy to improve feed efficiency and milk yield. However, the effects of YE on dairy cow performance remain inconsistent across studies, leaving the underlying mechanisms unclear. Hence, the primary aim of this study was to investigate the impact of YE supplementation on lactation performance, ruminal microbiota composition and fermentation patterns, as well as serum antioxidant capacity and immune functions in dairy cows. RESULTS: Supplementation with YE (20 g/d/head) resulted in enhancements in dairy cow's dry matter intake (DMI) (P = 0.016), as well as increased yields of milk (P = 0.002) and its components, including solids (P = 0.003), fat (P = 0.014), protein (P = 0.002), and lactose (P = 0.001) yields. The addition of YE led to significant increases in the concentrations of ammonia nitrogen (NH3-N) (P = 0.023), acetate (P = 0.005), propionate (P = 0.025), valerate (P = 0.003), and total volatile fatty acids (VFAs) (P < 0.001) in rumen fermentation parameters. The analysis of 16s rRNA gene sequencing data revealed that the administration of YE resulted in a rise in the relative abundances of three primary genera including Ruminococcus_2 (P = 0.010), Rikenellaceae_RC9_gut_group (P = 0.009), and Ruminococcaceae_NK4A214_group (P = 0.054) at the genus level. Furthermore, this increase was accompanied with an enriched pathway related to amino acid metabolism. Additionally, enhanced serum antioxidative (P < 0.05) and immune functionalities (P < 0.05) were also observed in the YE group. CONCLUSIONS: In addition to improving milk performance, YE supplementation also induced changes in ruminal bacterial community composition and fermentation, while enhancing serum antioxidative and immunological responses during the mid-lactation stage. These findings suggest that YE may exert beneficial effects on both rumen and blood metabolism in mid-lactation dairy cows.

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

BACKGROUND: 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. RESULTS: 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). CONCLUSION: 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.

High-energy diet modify rumen microbial composition and microbial energy metabolism pattern in fattening sheep.

Higher dietary energy is often used to achieve better animal performance in mutton sheep production. Notably, changing the diet formula affects rumen fermentation and the microbiota of ruminants. In this study, we investigated the effect of dietary energy on rumen fermentation and ruminal microbiota in fattening sheep. Fifteen 2-month-old white-headed Suffolk sheep (♂) × Hu sheep (♀) crossbred lambs were randomly divided into three treatments based on the dietary energy of the feeds fed: 8.67 MJ/kg (Low energy (LE); n = 5), 10.38 MJ/kg (standard energy (CON); n = 5), and 12.31 MJ/kg (high energy (HE); n = 5) groups. After 70 days of feeding, sheep were slaughtered and the ruminal fluids were collected and analyzed to determine fermentation parameters. Microbiota was determined using metagenomics sequencing. Notably, the microbial cell protein (MCP) and butyric acid concentrations were significantly high in the HE group. Metagenomic sequencing revealed that ACE and Chao indexes of the HE group were significantly decreased. Four genera among the major classified taxa across all the kingdoms differed in relative abundance in the three dietary energy levels. The relative abundances of Prevotella_brevis, Succiniclasticum_ruminis, Prevotellace-ae_bacterium, and Lachnospiraceae_bacterium were significantly correlated with rumen fermentation. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis further revealed that a high-energy diet increased lipid metabolism of microbiota. The Carbohydrate Active enzymes (CAZy) gene, which participates in energy metabolism, was upregulated, while genes regulating plant cell wall degradation were downregulated in the HE group. These results suggest that a high-energy diet had minimal influence on the rumen fermentation pattern but altered the composition of the rumen microbiota, enhancing microbial lipid metabolism and limiting crude fiber metabolism. The findings of this study provide scientific evidence of the effect of dietary energy on ruminant fermentation and fattening sheep production.

Effect of whole oilseeds in the diet on bacterial diversity in the solid fraction of the ruminal content of steers.

Our hypothesis was that different whole oilseeds included in the diet for steers confined could alter the diversity of rumen bacteria compared to a diet without oilseeds or an exclusively forage diet. It was aimed to evaluate the effects of oilseeds inclusion in the diet on bacterial diversity in the solid fraction of the ruminal content of steers, by gene sequences of the conserved 16S rDNA region. Six crossbred steers castrated males, fitted with ruminal cannula were used in a 6 × 6 Latin square design, using 21-day period. At the start of the experiment, the live weight of the animals averaged 416 ± 9.7 kg (mean ± SD). A total of 2,180,562 16S rDNA sequences were generated for the Bacteria domain by MiSeq sequencing. The bacterial diversity was composed of 24 bacterial phyla, with the most abundant being Firmicutes, Bacteroidetes, and Proteobacteria. Other phyla with less diversity were also identified including Eurychaeota, Tenericutes, SR1 Absconditalbacteria, Synergistetes, Actinobacteria, Saccharibacteria, Elusimicrobia, Cyanobacteria, Verrucomicrobia, Fusobacteria, Lentisphaerae. The similarity in the bacterial community averaged 50% for all the experimental diets. Steers-fed corn silage exhibited a great diversity of bacteria of the Firmicutes phylum. The steers-fed oilseeds in the diet had a great diversity of bacteria from the phylum Bacteroidetes and Proteobacteria. The inclusion of whole oilseeds in the steer diets can alter the rumen bacteria population by up to 50% of total diversity.

Sex differences in rumen fermentation and microbiota of Tibetan goat.

BACKGROUND: The gut microbiota play an important role in maintaining host metabolism, the immune system and health, while sex, genotype, diet and health have specific effects on the composition of the gut microbiota. Therefore, to explore the sex differences in the structure and function of rumen microbiota in Tibetan goats, herein we analyzed sex differences in rumen fermentation parameters, rumen microbiota and the expression of genes related to VFA transport in Tibetan goats. RESULTS: The results showed that the contents of acetic acid and propionic acid in the rumen of TGM (Tibetan goat male) were significantly higher than those in TGFm (Tibetan goat female) (P < 0.05), and total VFAs was significantly higher in TGM than TGFm (P < 0.05). Expression of the VFA transport-related genes DRA, AE2, MCT-1, NHE1, and NHE2 in the rumen epithelium of TGFm was significantly higher than that in TGM. Analysis of the composition and structure of the rumen microbiota revealed significant sex differences. At the phylum level, Firmicutes and Bacteroidetes were the dominant phyla in Tibetan goats. In addition, Fibrobacteres and Spirochaetes had significantly greater relative abundances in TGFm than in TGM (P < 0.05). At the genus level, the relative abundance of Fibrobacter, Ruminococcus_1 and Pyramidobacter was significantly higher in TGFm than in TGM (P < 0.05). The functional prediction results showed that replication, recombination and repair, RNA processing and modification were mainly enriched in TGFm (P < 0.05). CONCLUSIONS: Correlation analysis revealed significant associations of some rumen microbiota with the fermentation product VFAs and VFA transport-related genes. We concluded that yearling TGM and TGFm have distinct fermentation and metabolism abilities when adapting to the plateau environment, which provides a certain sex reference basis for Tibetan goat adaptation to the plateau environment.

Evaluation of gallnut tannin and Lactobacillus plantarum as natural modifiers for alfalfa silage: Ensiling characteristics, in vitro ruminal methane production, fermentation profile and microbiota.

AIMS: The potential of gallnut tannin (GT) and Lactobacillus plantarum (LP) on fermentation characteristics, in vitro ruminal methane (CH4 ) production and microbiota of alfalfa silage was investigated. METHODS AND RESULTS: Alfalfa was ensiled with GT (20 and 50 g kg-1 dry matter [DM]) and LP (3 × 108  CFU per gram fresh matter) alone or in combination for 60 days. The GT and LP alone or in combination decreased DM losses, pH and non-protein nitrogen contents of alfalfa silage. All additive treatments decreased ruminal CH4 production, and increased propionic acid molar proportions and Fibrobacter succinogenes numbers. The LP treatment increased nutrient degradation, cellobiase, pectinase and protease activities, and Prevotella ruminicola abundance, whereas high-dose GT treatment inhibited these variables. Importantly, LP together with GT alleviated the adverse effects of high-dose GT supply alone by enhancing pectinase and protease activities as well as Rumincoccus flavefaciens and P. ruminicola growth. CONCLUSIONS: Combination of GT and LP can be used as an efficient additive to improve silage quality and utilization by ruminants. SIGNIFICANCE AND IMPACT OF THE STUDY: Using GT-LP combination has practical implications, particularly concerning effects of tannins on ruminal CH4 mitigation, which may alleviate inhibitory effects of tannins on feed digestion through modulating ruminal microbiota.

Residual feed intake in peripartal dairy cows is associated with differences in milk fat yield, ruminal bacteria, biopolymer hydrolyzing enzymes, and circulating biomarkers of immunometabolism.

Residual feed intake (RFI) measures feed efficiency independent of milk production level, and is typically calculated using data past peak lactation. In the current study, we retrospectively classified multiparous Holstein cows (n = 320) from 5 of our published studies into most feed-efficient (M-eff) or least feed-efficient (L-eff) groups using performance data collected during the peripartal period. Objectives were to assess differences in profiles of plasma biomarkers of immunometabolism, relative abundance of key ruminal bacteria, and activities of digestive enzymes in ruminal digesta between M-eff and L-eff cows. Individual data from cows with ad libitum access to a total mixed ration from d -28 to d +28 relative to calving were used. A linear regression model including dry matter intake (DMI), energy-corrected milk (ECM), changes in body weight (BW), and metabolic BW was used to classify cows based on RFI divergence into L-eff (n = 158) and M-eff (n = 162). Plasma collected from the coccygeal vessel at various times around parturition (L-eff = 60 cows; M-eff = 47 cows) was used for analyses of 30 biomarkers of immunometabolism. Ruminal digesta collected via esophageal tube (L-eff = 19 cows; M-eff = 29 cows) was used for DNA extraction and assessment of relative abundance (%) of 17 major bacteria using real-time PCR, as well as activity of cellulase, amylase, xylanase, and protease. The UNIVARIATE procedure of SAS 9.4 (SAS Institute Inc.) was used for analyses of RFI coefficients. The MIXED procedure of SAS was used for repeated measures analysis of performance, milk yield and composition, plasma immunometabolic biomarkers, ruminal bacteria, and enzyme activities. The M-eff cows consumed less DMI during the peripartal period compared with L-eff cows. In the larger cohort of cows, despite greater overall BW for M-eff cows especially in the prepartum (788 vs. 764 kg), no difference in body condition score was detected due to RFI or the interaction of RFI × time. Milk fat content (4.14 vs. 3.75 ± 0.06%) and milk fat yield (1.75 vs. 1.62 ± 0.04 kg) were greater in M-eff cows. Although cumulative ECM yield did not differ due to RFI (1,138 vs. 1,091 ± 21 kg), an RFI × time interaction due to greater ECM yield was found in M-eff cows. Among plasma biomarkers studied, concentrations of nonesterified fatty acids, ß-hydroxybutyrate, bilirubin, ceruloplasmin, haptoglobin, myeloperoxidase, and reactive oxygen metabolites were overall greater, and glucose, paraoxonase, and IL-6 were lower in M-eff compared with L-eff cows. Among bacteria studied, abundance of Ruminobacter amylophilus and Prevotella ruminicola were more than 2-fold greater in M-eff cows. Despite lower ruminal activity of amylase in M-eff cows in the prepartum, regardless of RFI, we observed a marked linear increase after calving in amylase, cellulase, and xylanase activities. Protease activity did not differ due to RFI, time, or RFI × time. Despite greater concentrations of biomarkers reflective of negative energy balance and inflammation, higher feed efficiency measured as RFI in peripartal dairy cows might be associated with shifts in ruminal bacteria and amylase enzyme activity. Further studies could help address such factors, including the roles of the liver and the mammary gland.

Supplementation of Ascophyllum nodosum meal and monensin: Effects on diversity and relative abundance of ruminal bacterial taxa and the metabolism of iodine and arsenic in lactating dairy cows.

Previous research has shown that the brown seaweed Ascophyllum nodosum (ASCO) has antimicrobial and antioxidant properties and also increases milk I concentration. We aimed to investigate the effects of supplementing ASCO meal or monensin (MON) on ruminal fermentation, diversity and relative abundance of ruminal bacterial taxa, metabolism of I and As, and blood concentrations of thyroid hormones, antioxidant enzymes, and cortisol in lactating dairy cows. Five multiparous ruminally cannulated Jersey cows averaging (mean ± standard deviation) 102 ± 15 d in milk and 450 ± 33 kg of body weight at the beginning of the study were used in a Latin square design with 28-d periods (21 d for diet adaptation and 7 d for data and sample collection). Cows were fed ad libitum a basal diet containing (dry matter basis) 65% forage as haylage and corn silage and 35% concentrate and were randomly assigned to 1 of the following 5 dietary treatments: 0, 57, 113, or 170 g/d of ASCO meal, or 300 mg/d of MON. Supplements were placed directly into the rumen once daily after the morning feeding. Diets had no effect on ruminal pH and NH3-N concentration, which averaged 6.02 and 6.86 mg/dL, respectively. Total volatile fatty acid concentration decreased linearly in cows fed incremental amounts of ASCO meal. Supplementation with ASCO meal did not change the ruminal molar proportions of volatile fatty acids apart from butyrate, which responded quadratically with the lowest values observed at 56 and 113 g/d of ASCO supplementation. Compared with the control diet or diets containing ASCO meal, cows fed MON showed greater molar proportion of propionate. Diets did not affect the α diversity indices Shannon, Simpson, and Fisher for ruminal bacteria. However, feeding incremental levels of ASCO meal linearly decreased the relative abundance of Tenericutes in ruminal fluid. Monensin increased the relative abundance of the CAG:352 bacterial genus in ruminal fluid compared with the control diet. Linear increases in response to ASCO meal supplementation were observed for the concentrations and output of I in serum, milk, urine, and feces. Fecal excretion of As increased linearly in cows fed varying amounts of ASCO meal, but ASCO did not affect the concentration and secretion of As in milk. The plasma activities of the antioxidant enzymes and the serum concentrations of thyroid hormones did not change. In contrast, circulating cortisol decreased linearly in diets containing ASCO meal. The apparent total-tract digestibilities of dry matter, organic matter, and crude protein increased linearly with ASCO meal, but those of neutral and acid detergent fiber were not affected. In summary, feeding incremental amounts of ASCO meal decreased serum cortisol concentration, and increased I concentrations and output in serum, milk, feces, and urine.

Microbial mechanisms of using feruloyl esterase-producing Lactobacillus plantarum A1 and grape pomace to improve fermentation quality and mitigate ruminal methane emission of ensiled alfalfa for cleaner animal production.

This study was conducted to investigate the influence of feruloyl esterase-producing Lactobacillus plantarum A1 (Lp A1) and grape pomace (GP) alone, or in combination (LG) on ensiling characteristics and bacterial community, in vitro ruminal fermentation, methane (CH4) emission, and the microbiota of ensiled alfalfa. Alfalfa at 42% dry matter (DM) was treated in a 2 × 2 factorial design: with the application of Lp A1 at 0 (control) or 1 × 106 cfu/g of fresh forage, and GP at 0 or 5% of fresh forage. After 60 d of ensiling, a decrease in nonprotein nitrogen (NPN) was observed in GP treated silage. Lp A1 inoculated silage had a lower fiber content than silages without Lp A1. The lowest NPN was found in silage treated with LG, and an obvious increase in the relative abundance of Lactobacillus paracasei was detected in silages treated with Lp A1 and LG, respectively. In vitro ruminal experiments indicated that, although the application of GP deceased ruminal total gas, CH4 production, nitrogen degradation and the number of methanogenic archaea in alfalfa silage, it also reduced silage DM digestibility. In contrast, inoculation with Lp A1 not only increased DM digestibility and populations of ruminal Ruminococcus flavefaciens and fungi, but also improved ruminal total gas and CH4 production. As expected, LG treatment decreased alfalfa silage ruminal total gas and CH4 production relative to Lp A1 treatment alone, and increased silage DM digestibility compared with GP treated silage. In conclusion, the application of LG before ensiling alfalfa, balanced silage proteolysis, feed digestibility, and CH4 emission, and could be a promising strategy for using food industry by-products to produce a nutritional and environmentally-friendly legume silage that will mitigate N and greenhouse gas emissions from ruminants.

Bacterial species in the ruminal content of steers fed oilseeds in the diet.

The aim of this study was to evaluate bacterial species and diversity of methanogenic Archaea in the solid fraction of the ruminal content, through the gene sequences of the conserved 16S rDNA region, in response to the following diets: canola, cottonseed, sunflower, soybean, corn silage, and control diet. Six rumen-fistulated crossbred steers, with body weight (BW) of 416.33 ± 93.30 kg, were distributed in a 6 × 6 Latin square design. Regardless of the diet provided, amylolytic, proteolytic, and lactic bacteria were identified in the rumen fluid. Cellulolytic bacteria were predominant for all diets, reaching 47.75% of operational taxonomic units (OTUs) in animals fed with the cottonseed diet. Amylolytic bacteria reach 62.51% of OTU in animal fed sunflower diet, while proteolytic bacteria correspond to 65.96% of OTU in this same diet. Also, Megasphaera elsdenii bacterium was identified for all diets, with a greater percentage of OTU in steers fed the cottonseed diet. The diversity analysis of the species identified the methanogenic Archaea Methanobrevibacter ruminantium in all diets. We conclude that the control and corn silage diets have the most similar bacterial flora; diets with oilseeds had 47.5% similarity in rumen flora bacteria species. Animals fed with soybean showed a reduced number of methanogenic Archaea in the rumen content, which could be an alternative feed for cattle due to their low potential for energy losses with the production of methane.

Astragalus root extract improved average daily gain, immunity, antioxidant status and ruminal microbiota of early weaned yak calves.

BACKGROUND: Early weaning in yak calves is being attempted to improve yak reproduction rate. However, this has to be done with caution because of the high mortality rate of calves due to the lack of nutrients and the harsh environmental conditions. Twenty-four weaned male yak calves were used in a 60 day feeding trial in which astragalus root extract (ARE) was supplemented. They were assigned randomly to one of four dietary treatments (n = six per treatment) that differed in ARE level: 0 g kg-1 (control), ARE0 ; 20 g kg-1 , ARE20 ; 50 g kg-1 , ARE50; and 80 g kg-1 dry matter intake (DMI), ARE80 . RESULTS: Final bodyweight and average daily gain (ADG) were significantly higher and the DMI/ADG ratio was significantly lower in calves with ARE supplementation than control (ARE0 ) calves. Ruminal concentrations of acetate and propionate and serum concentration of superoxide dismutase in ARE80 calves were higher than in the other groups and serum concentration of insulin was higher in ARE80 calves than in ARE20 calves. Serum immunoglobulin G (IgG) and interleukin-2 (IL-2) concentrations in ARE-fed calves were higher than in controls. Serum tumor necrosis factor (TNF-α) concentration was higher in ARE50 and ARE80 groups than ARE0 calves and serum interleukin-6 (IL-6) concentration was higher in ARE80 than in ARE0 calves. Serum immunoglobulin A (IgA), IgG and immunoglobulin M (IgM) concentrations increased with age in ARE-fed calves. ARE supplementation increased the abundance of fiber degrading bacteria. CONCLUSION: ARE at a dosage of 5% to 8% DMI can be supplemented to early weaned yak calves to improve growth performance, antioxidant capacity and immunity. © 2020 Society of Chemical Industry.

Effect of replacing conventional feeds with tropical agricultural by-products on the growth performance, nutrient digestibility and ruminal microbiota of water buffaloes.

This study investigated the effect of replacing corn grain, soybean meal and wheat bran with tropical agricultural by-products, such as palm kernel cake (PKC), cassava residue and dried distiller's grain with solubles (DDGS), on the dry matter intake (DMI), growth performance, apparent nutrient digestibility, ruminal short-chain fatty acids (SCFA) and ruminal microbial communities of water buffaloes. Thirty healthy 15-month-old crossbred water buffaloes with a similar initial body weight of 353.1 ± 23.7 kg were randomly allocated into three dietary experimental groups, and they were fed with same forage but three different concentrates for 50 days fattening. The dietary treatments were as following: typical concentrate (TC, 65% corn + 15% wheat bran + 15% soybean meal), partial replacement concentrate Ⅰ (PRC I, 50% corn + 22.5% corn gluten + 22.5% PKC) and partial replacement concentrate Ⅱ (PRC II, 50% corn + 22.5% cassava residue + 22.5% DDGS). The results showed that the average daily gain of the PRC II group was the highest, and the DMI, acid detergent fibre digestibility and neutral detergent fibre digestibility value of the three groups were different and in the following order: PRC II group > TC group > PRC I group. The crude protein digestibility of PRC II was higher than that of the TC and PRC I groups (p < .05). The ruminal concentrations of total SCFA, acetate, propionate and butyrate of TC group were higher than the other two groups (p < .05). The PRC I group had the highest Bacteroidetes-to-Firmicutes ratio (B/F) and relative abundance of the genus Prevotella, while the PRC II group had the lowest B/F and relative abundance of Prevotella. In conclusion, using PKC and corn gluten to completely replace common feed ingredients in the buffalo concentrate ration decreased, while using cassava residue and DDGS increased animal growth performance, mainly due to the different combination influenced nutrient digestibility and ruminal microbial community composition was shifted.

Effect of dietary concentrate to forage ratios on ruminal bacterial and anaerobic fungal populations of cashmere goats.

The rumen contains a highly complex microbial ecosystem that plays an important role in converting solar energy in plants into nutrients for ruminants and generates animal food products, such as meat and milk for humans. Therefore, understanding the effect of the dietary concentrate to forage (C:F) ratio on ruminal microbiota is of great significance for the growth and development of ruminants. In this study, changes in the ruminal bacterial and anaerobic fungal populations of Shaanbei white-cashmere (SWC) goats that were reared under different dietary C:F ratios were evaluated by high-throughput sequencing analysis. It was found that dietary C:F ratio has a significant impact on the composition of the ruminal bacteria in SWC goats. The levels of Actinobacteria and Proteobacteria were significantly increased (P < 0.05), whereas the level of Bacteroidetes was significantly decreased when the proportion of dietary concentrate was increased (P < 0.05); as the proportion of dietary concentrate increased, Prevotella, Selenomonas, and Treponema were significantly increased (P < 0.05), whereas Oscillospira and Succiniclasticum were significantly reduced (P < 0.05). Furthermore, different dietary C:F ratios significantly affected the composition of anaerobic fungi in SWC goats. As the proportion of dietary concentrate increased, Ascomycota, Basidiomycota, and Zygomycota were significantly increased (P < 0.05), while Neocallimastigomycota was significantly reduced (P < 0.05); the levels of Alternaria, Aspergillus, Neocallimastix, Orpinomyces, Piromyces, and Stachybotrys were significantly increased, while those of Candida, Penicillium, and Trichosporon were significantly decreased when the proportion of dietary concentrate increased (P < 0.05). These findings will help us to better understand the changes in ruminal bacterial and anaerobic fungal populations of SWC goats under different dietary C:F ratios, which could provide a theoretical basis for microecological regulation of SWC goats.

Milk fat depression in dairy ewes fed fish oil: Might differences in rumen biohydrogenation, fermentation, or bacterial community explain the individual variation?

Dairy ewes show large individual variation in the extent of diet-induced milk fat depression (MFD) but reasons behind this variability remain uncertain. Previous results offered no convincing support for these differences being related to relevant changes in the milk fatty acid (FA) profile, including potentially antilipogenic FA, or in the transcript abundance of candidate genes involved in mammary lipogenesis. Therefore, we hypothesized that alterations in the processes of rumen biohydrogenation and fermentation, as well as in the bacterial community structure, might account for individual variation in fish oil-induced MFD severity. To test this explanation, 15 ewes received a total mixed ration without lipid supplementation (control; n = 5) or supplemented with 20 g of fish oil/kg of dry matter [10 animals divided into those showing a strong (RESPON+; -25.4%; n = 5) or a mild (RESPON-; -7.7%; n = 5) decrease in milk fat concentration] for 5 wk. Rumen fermentation parameters, biohydrogenation metabolites, and bacterial structure and diversity were analyzed in rumen samples collected before and after treatments. Although the fish oil supplementation increased the concentration of demonstrated or putative antilipogenic FA (e.g., cis-9 16:1, cis-11 18:1, or trans-10,cis-12 CLA), surprisingly, none of them differed significantly in relation to the extent of MFD (i.e., between RESPON- and RESPON+), and this was the case only for a few minor FA (e.g., cis-6+7 16:1 or 17:0 anteiso). Changes in total volatile FA, acetate, and propionate concentrations were associated with MFD severity, with higher decreases in more susceptible animals. Individual responses were not related to shifts in rumen bacterial structure but some terminal restriction fragments compatible with Clostridiales, Ruminococcaceae, Lachnospiraceae, and Succiniclasticum showed greater abundances in RESPON-, whereas some others that may correspond to Prevotella, Mogibacterium, and Quinella-related spp. were more abundant in RESPON+. Overall, the results suggest that individual variation in MFD severity in dairy ewes fed fish oil cannot be fully explained by differences in the processes of rumen biohydrogenation and fermentation or in the bacterial community, and further research would be necessary to elucidate the large variability in the responsiveness to MFD-inducing marine lipids.

Graded substitution of grains with bakery by-products modulates ruminal fermentation, nutrient degradation, and microbial community composition in vitro.

A new segment of feed industry based on bakery by-products (BBP) has emerged. Yet, information is lacking regarding the effects of inclusion of BBP in ruminant diets on ruminal fermentation and microbiota. Therefore, the aim of this study was to evaluate the effect of the gradual replacement of grains by BBP on ruminal fermentation, nutrient degradation, and microbial community composition using the rumen-simulation technique. All diets consisted of hay and concentrate mixture with a ratio of 42:58 (dry matter basis), but differed in the concentrate composition with either 45% cereal grains or BBP, whereby 15, 30, or 45% of BBP were used in place of cereal grains. The inclusion of increasing levels of BBP in the diet linearly enhanced ruminal degradation of starch from 84% (control) to 96% (45% BBP), while decreasing degradation of crude protein and fiber. The formation of methane was lowered in the 45% BBP diet compared with all other diets. Whereas the ammonia concentration was similar in the control and 15% BBP, a significant decrease was found in 30% BBP (-23%) and 45% BBP (-33%). Also, BBP feeding shifted fermentation profile toward propionate at the expense of acetate. Moreover, isobutyrate linearly decreased with increasing BBP inclusion. Bacterial 16S rRNA Illumina MiSeq (Microsynth AG, Balach, Switzerland) sequencing revealed a decreased microbial diversity for the 45% BBP diet. Furthermore, the replacement of cereal grains with BBP went along with an increased abundance of the genera Prevotella, Roseburia, and Megasphaera, while decreasing Butyrivibrio and several OTU belonging to Ruminococcaceae. In conclusion, the inclusion of BBP at up to 30% of the dry matter had no detrimental effects on pH, fiber degradability, and microbial diversity, and enhanced propionate production. However, a higher replacement level (45%) impaired ruminal fermentation traits and fiber degradation and is not recommended.

Changes in Microbiota in Rumen Digesta and Feces Due to a Grain-Based Subacute Ruminal Acidosis (SARA) Challenge.

The effects of a grain-based subacute ruminal acidosis (SARA) challenge on bacteria in the rumen and feces of lactating dairy cows were determined. Six lactating, rumen-cannulated Danish Holstein cows were used in a cross-over study with two periods. Periods included two cows on a control diet and two cows on a SARA challenge. The control diet was a total mixed ration containing 45.5% dry matter (DM), 43.8% DM neutral detergent fiber, and 19.6% DM starch. The SARA challenge was conducted by gradually substituting the control diet with pellets containing 50% wheat and 50% barley over 3 days to reach a diet containing 55.6% DM, 31.3% DM neutral detergent fiber, and 31.8% DM starch, which was fed for four more days. Rumen fluid samples were collected at day 7 and 10 of experimental periods. Feces samples were collected on days 8 and 10 of these periods. Extracted DNA from the rumen and feces samples was analyzed to assess their bacterial communities using MiSeq Illumina sequencing of the V4 region of the 16S rRNA gene. The induction of SARA reduced the richness, diversity, and stability of bacterial communities and resulted in distinctly different microbiota in the rumen and feces. Bacteroidetes and Firmicutes were the most abundant phyla and, combined, they represented 76.9 and 94.4% of the bacterial community in the rumen fluid and the feces, respectively. Only the relative abundance of Firmicutes in the rumen was increased by the SARA challenge. In rumen fluid and feces, the abundances of nine out of the 90 and 25 out of the 89 taxa, respectively, were affected by the challenge. Hence, SARA challenge altered the composition of the bacterial community at the lower taxonomical level in the feces and therefore also likely in the hindgut, as well as in the rumen. However, only reductions in the bacterial richness and diversity in the rumen fluid and feces were in agreement with those of other studies and had a biological basis. Although the composition of the bacterial community of the feces was affected by the SARA challenge, bacterial taxa in the feces that can be used for accurate and non-invasive diagnosis of SARA could not be identified.

Effects of co-inoculating rice straw with ruminal microbiota and anaerobic sludge: digestion performance and spatial distribution of microbial communities.

Ruminal microbiota (RM) were co-inoculated with anaerobic sludge (AS) at different ratios to study the digestion of rice straw in batch experiments. The CH4 yield reached 273.64 mL/g volatile solid (VS) at a co-inoculum ratio of 1:1. The xylanase and cellulase activities were 198.88-212.88 and 24.51-29.08 U/mL in co-inoculated samples, respectively, and were significantly different compared to the results for single inoculum (p < 0.05). Higher ratios of AS enhanced acetoclastic methanogenesis, and propionate accumulation could be the main reason for the longer lag phase observed in samples with a higher RM ratio. The microbial compositions were clearly altered after digestion. Fibrobacter, Ruminococcus and Butyrivibrio from the rumen did not settle in the co-inoculated system, whereas Clostridiales members became the main polysaccharide degraders. Microbial interactions involving hydrolytic bacteria and acetoclastic methanogens in the residue were considered to be significant for hydrolysis activities and methane production. Syntrophy involving propionate oxidizers with associated methanogens occurred in the liquid phase. Our findings provide a better understanding of the anaerobic digestion of rice straw that is driven by specific microbial populations.

Rumen microbial and fermentation characteristics are affected differently by acarbose addition during two nutritional types of simulated severe subacute ruminal acidosis in vitro.

Little information is available on whether or not the effect of an alpha-glucosidase inhibitor on the prevention of ruminal acidosis is influenced by the type of diet during ruminant feeding. This study was conducted to explore the effect of acarbose addition on the prevention of severe subacute ruminal acidosis induced by either cracked wheat or beet pulp in vitro. Cracked wheat and beet pulp were fermented in vitro by rumen microorganisms obtained from three dairy cows. When cracked wheat was used as the substrate and fermented for 24 h, compared with the control, acarbose addition decreased the concentrations of acetate, propionate, butyrate, total volatile fatty acids, and lactate (P < 0.05), while linearly increased the ratio of acetate to propionate, pH value, and the ammonia-nitrogen level (P < 0.05). Applying Illumina MiSeq sequencing of a fragment of the 16S rRNA gene revealed that the relative abundance of Firmicutes and Bacteroidetes as well as the ACE (abundance-based coverage estimator) value, Chao 1 value, and Shannon index increased significantly (P < 0.05), while there was a significant reduction (P < 0.05) in the relative abundance of Tenericutes as well as Proteobacteria after adding acarbose compared to the control. On the other hand, when beet pulp was used as the substrate, acarbose addition had no significant effects (P > 0.05) on the fermentation parameters and the Chao 1 value, the Shannon index, and the proportion of Firmicutes and Bacteroidetes. In general, these findings indicate that acarbose had more effects on ruminal fermentation when wheat was used as the substrate, whereas it exhibited little effect on ruminal fermentation when beet pulp was used as the substrate.

Sheep fed with banana leaf hay reduce ruminal protozoa population.

A ciliate protozoa suppression can reduce methane production increasing the energy efficiency utilization by ruminants. The physicochemical characteristics of rumen fluid and the profile of the rumen protozoa populations were evaluated for sheep fed banana leaf hay in replacement of the Cynodon dactylon cv. vaqueiro hay. A total of 30 male sheep were raised in intensive system during 15 days of adaptation and 63 days of experimental period. The animals were distributed in a completely randomized design that included six replicates of five treatments with replacement levels (0, 25, 50, 75, and 100%) of the grass vaquero for the banana leaf hay. Samples of fluid were collected directly from the rumen with sterile catheters. Color, odor, viscosity, and the methylene blue reduction potential (MBRP) were evaluated and pH estimated using a digital potentiometer. After decimal dilutions, counts of genus protozoa were performed in Sedgewick Rafter chambers. The averages of pH, MBRP, color, odor, and viscosity were not influenced by the inclusion of the banana leaf hay. However, the total number of protozoa and Entodinium spp. population significantly decreased at 75 and 100% inclusions of banana leaf hay as roughage.

Fortification of dried distillers grains plus solubles with grape seed meal in the diet modulates methane mitigation and rumen microbiota in Rusitec.

The role of dried distillers grains plus solubles (DDGS) and associative effects of different levels of grape seed meal (GSM) fortified in DDGS, used as both protein and energy sources in the diet, on ruminal fermentation and microbiota were investigated using rumen-simulation technique. All diets consisted of hay and concentrate mixture with a ratio of 48:52 [dry matter (DM) basis], but were different in the concentrate composition. The control diet contained soybean meal (13.5% of diet DM) and barley grain (37%), whereas DDGS treatments, unfortified DDGS (19.5% of diet DM), or DDGS fortified with GSM, either at 1, 5, 10, or 20% were used entirely in place of soybean meal and part of barley grain at a 19.5 to 25% inclusion level. All diets had similar DM, organic matter, and crude protein contents, but consisted of increasing neutral detergent fiber and decreasing nonfiber carbohydrates levels with DDGS-GSM inclusion. Compared with the soy-based control diet, the unfortified DDGS treatment elevated ammonia concentration (19.1%) of rumen fluid associated with greater crude protein degradation (~19.5%). Methane formation decreased with increasing GSM fortification levels (≥ 5%) in DDGS by which the methane concentration significantly decreased by 18.9 to 23.4 and 12.8 to 17.6% compared with control and unfortified DDGS, respectively. Compared with control, unfortified DDGS decreased butyrate proportion, and GSM fortification in the diet further decreased this variable. The proportions of genus Prevotella and Clostridium cluster XIVa were enhanced by the presence of DDGS without any associative effect of GSM fortification. The abundance of methanogenic archaea was similar, but their composition differed among treatments; whereas Methanosphaera spp. remained unchanged, proportion of Methanobrevibacter spp. decreased in DDGS-based diets, being the lowest with 20% GSM inclusion. The abundance of Ruminococcus flavefaciens, anaerobic fungi, and protozoa were decreased by the GSM inclusion. As revealed by principal component analysis, these variables were the microorganisms associated with the methane formation. Grape seed meal fortification level in the diet decreased DM and organic matter degradation, but this effect was more related to a depression of nonfiber carbohydrates degradation. It can be concluded that DDGS fortified with GSM can favorably modulate ruminal fermentation.