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Fungal probiotics have the potential as feed additives, but less has been explored in ruminant feed up to date. This study aimed to determine the effect of compound probiotics (CPs) with Aspergillus oryzae 1, Aspergillus oryzae 2 and Candida utilis on Hu sheep's growth performance, rumen fermentation and microbiota. A total of 120 male Hu sheep, aged 2 months and with the body weight of 16.95 ± 0.65 kg were divided into 4 groups. Each group consisted of 5 replicates, with 6 sheep per replicate. Group A was the control group fed with the basal diet. Group B, C and D was supplemented with the basal diet by adding 400, 800 and 1,200 grams per ton (g/t) CPs, respectively. The feeding trial lasted for 60 days after a 10-day adaptation period. The results showed that the average daily gain (ADG) of sheep in the CPs groups were significantly higher, the feed/gain were significantly lower than those in group A in the later stage and the overall period. The addition of CPs increased the economic benefit. The levels of CD4+ and the CD4+/CD8+ ratio in the CPs groups were higher than those in Group A. The levels of GSH, IgG, IL-2, IL-6, and IFN- γ in group C were significantly elevated compared with group A. Group B showed a significant increase in rumen NH3-N and cellulase activity. There was no difference in VFAs content between group A and group B, however, with the increasing addition of CPs, the butyric acid and isobutyric acid content tended to decrease. The rumen microbiota analysis indicated that the CPs addition increased the Firmicutes and Proteobacteria abundances, decreased the Bacteroidetes abundance. The correlation analysis showed that Prevotella was negatively correlated with ADG, and the addition of 400 CPs in group B reduced Prevotella 's relative abundance, indicating CPs increased sheep growth by decreasing Prevotella abundance. The CPs addition reduced caspase-3, NF-κB and TNF- α expression in liver, jejunum and rumen tissues. In conclusion, the addition of CPs increased the sheep production performance, reduced inflammation, improved rumen and intestinal health. Considering the above points and economic benefits, the optimal addition of CPs as an additive for Hu sheep is 800 g/t. [ABSTRACT FROM AUTHOR]

Dietary crude protein concentration oscillation can improve the nitrogen utilization efficiency of ruminants. However, little is known about the dynamic changes in microbiota and fermentation in the rumen of calves during the oscillation period. In this study, six calves were fed an oscillating diet at 2-day intervals, including a high-protein diet (HP) and a low-protein diet (LP). The rumen fermentation parameters, plasma urea-N concentration, and rumen bacterial diversity were characterized throughout the oscillation period. The concentrations of volatile fatty acids, NH3-N, and plasma urea-N in rumen changed significantly with an oscillating diet. The abundance of Prevotella_1, Selenomonadales, Succiniclasticum, Clostridiales, Ruminococcaceae, Lachnospiraceae, and Rikenellaceae_RC9_gut_group showed significant changes with diet. Prevotella_1 was positively correlated, and Lachnospiraceae_AC2044_group and Saccharofermentans were negatively correlated with NH3-N. The abundance of Amino Acid Metabolism, Metabolism of Other Amino Acids, and Glycan Biosynthesis and Metabolism pathways, annotated by bacterial functional genes, decreased when the diet changed from HP to LP. The abundance of the Carbohydrate Metabolism pathway increased after the two dietary changes. In conclusion, the plasma urea-N concentration was not as sensitive and quick to adapt to diet changes as the rumen fermentation parameters. Rumen bacteria were responsible for increasing the nitrogen utilization efficiency of calves fed an oscillating diet. [ABSTRACT FROM AUTHOR]

ABSTRACT: The nutritional components and quality of milk are influenced by the rumen microbiota and its metabolites at different lactation stages. Hence, rumen fluid and milk samples from 6 dairy cows fed the same diet were collected during peak lactation, early mid-lactation, and later mid-lactation. Untargeted metabolomics and 16S rRNA sequencing were applied for analyzing milk and rumen metabolites, as well as rumen microbial composition, respectively. The levels of lipid-related metabolites, l-glutamate, glucose-1-phosphate, and acetylphosphate in milk exhibited lactation-dependent attenuation. Maltol, N-acetyl-d-glucosamine, and choline, which are associated with milk flavor or coagulation properties, as well as l-valine, lansioside A, clitocine, and ginsenoside La, increased significantly in early mid-lactation and later mid-lactation, especially in later mid-lactation. The obvious increase in rumen microbial diversities (ACE and Shannon indices) were observed in early mid-lactation compared with peak lactation. Twenty-one differential bacterial genera of the rumen were identified, with Succinivibrionaceae_UCG-001, Candidatus Saccharimonas, Fibrobacter, and SP3-e08 being significantly enriched in peak lactation. Rikenellaceae_RC9_gut_group, Eubacterium_ruminantium_group, Lachnospira, Butyrivibrio, Eubacterium_hallii_group, and Schwartzia were most significantly enriched in early mid-lactation. In comparison, only 2 bacteria (unclassified_f__Prevotellaceae and Prevotellaceae_UCG-001) were enriched in later mid-lactation. For rumen metabolites, LysoPE(16:0), l-glutamate, and l-tyrosine had higher levels in peak lactation, whereas PE(17:0/0:0), PE(16:0/0:0), PS(18:1(9Z)/0:0), l-phenylalanine, dulcitol, 2-(methoxymethyl)furan, and 3-phenylpropyl acetate showed higher levels in early mid-lactation and later mid-lactation. Multiomics-integrated analysis revealed that a greater abundance of Fibrobacter contributed to phospholipid content in milk by increasing ruminal acetate, l-glutamate, and LysoPE(16:0). Prevotellaceae_UCG-001 and unclassified_f_Prevotellaceae provide substrates for milk metabolites of the same category by increasing ruminal l-phenylalanine and dulcitol contents. These results demonstrated that milk metabolomic fingerprints and critical functional metabolites during lactation, and the key bacteria in rumen related to them. These findings provide new insights into the development of functional dairy products.

Crop residues have shown promise as non-conventional feed sources to enhance animal health and growth. This study evaluated the effects of chili straw (CS) on rumen fermentation, meat quality, amino and fatty acid composition, and rumen microbial diversity in sheep. Fifty F1 Dorper×Hu lambs (29.58 ± 2.06 kg) were randomly assigned to five groups, fed pelleted feed with 0%, 5%, 10%, 15%, or 20% CS over a 63-day period, including a 7-day pre-test. Post-trial, rumen fluid was sampled to assess fermentation and microbial profiles, and slaughter performance and meat quality were evaluated. Key findings include: (1) No significant differences were observed in rumen pH, NH3-N, or acetic acid-to-propionic acid ratio across groups (P > 0.05). (2) Rumen microbial diversity indices did not vary significantly between groups (P > 0.05), though the relative abundance of Firmicutes and Proteobacteria increased, and Bacteroidota decreased in CS-fed groups, with specific genus-level changes. (3) Carcass weight decreased in the CS20% group (P < 0.01). (4) Cooking loss decreased in CS10%, 15%, and 20% groups (P < 0.05), and meat redness increased in CS15% and 20% groups (P < 0.01). (5) Saturated fatty acids decreased, while the PUFA/SFA ratio and amino acid profiles, including sulfur-containing amino acids (SAA), dibasic amino acids (DAA), essential amino acids (EAA), and total amino acids (TAA), increased with CS, with a significant rise in Gly content in the CS15% group (P < 0.05). In conclusion, incorporating CS into lamb diets can enhance meat quality without adversely affecting rumen fermentation, with recommended levels between 10% and 15%.

Abstract The Yunshang black goat is a renowned mutton specialist breed mainly originating from China that has excellent breeding ability with varying litter sizes. Litter size is an important factor in the economics of goat farming. However, ruminal microbiome structure might be directly or indirectly regulated by pregnancy-associated factors, including litter sizes. Therefore, the current experiment aimed to evaluate the association of different litter sizes (low versus high) with ruminal microbiome structure by 16S rRNA gene sequencing and metabolomic profiling of Yunshang black does. A total of twenty does of the Yunshang Black breed, approximately aged between 3 and 4 years, were grouped (n = 10 goats/group) into low (D-l) and high (D-h) litter groups according to their litter size (the lower group has ≤ 2 kids/litter and the high group has ≧ 3 kids/litter, respectively). All goats were sacrificed, and collected ruminal fluid samples were subjected to 16S rRNA sequencing and LC–MS/MC Analysis for ruminal microbiome and metabolomic profiling respectively. According to PCoA analysis, the ruminal microbiota was not significantly changed by the litter sizes among the groups. The Firmicutes and Bacteroidetes were the most dominant phyla, with an abundance of 55.34% and 39.62%, respectively. However, Ruminococcaceae_UCG-009, Sediminispirochaeta, and Paraprevotella were significantly increased in the D-h group, whereas Ruminococcaceae_UCG-010 and Howardella were found to be significantly decreased in the D-l group. The metabolic profiling analysis revealed that litter size impacts metabolites as 29 and 50 metabolites in positive and negative ionic modes respectively had significant differences in their regulation. From them, 16 and 24 metabolites of the D-h group were significantly down-regulated in the positive ionic mode, while 26 metabolites were up-regulated in the negative ionic mode for the same group. The most vibrant identified metabolites, including methyl linoleate, acetylursolic acid, O-desmethyl venlafaxine glucuronide, melanostatin, and arginyl-hydroxyproline, are involved in multiple biochemical processes relevant to rumen roles. The identified differential metabolites were significantly enriched in 12 different pathways including protein digestion and absorption, glycerophospholipid metabolism, regulation of lipolysis in adipocytes, and the mTOR signaling pathway. Spearman’s correlation coefficient analysis indicated that metabolites and microbial communities were tightly correlated and had significant differences between the D-l and D-h groups. Based on the results, the present study provides novel insights into the regulation mechanisms of the rumen microbiota and metabolomic profiles leading to different fertility in goats, which can give breeders some enlightenments to further improve the fertility of Yunshang Black goats.

The Yunshang black goat is a renowned mutton specialist breed mainly originating from China that has excellent breeding ability with varying litter sizes. Litter size is an important factor in the economics of goat farming. However, ruminal microbiome structure might be directly or indirectly regulated by pregnancy-associated factors, including litter sizes. Therefore, the current experiment aimed to evaluate the association of different litter sizes (low versus high) with ruminal microbiome structure by 16S rRNA gene sequencing and metabolomic profiling of Yunshang black does. A total of twenty does of the Yunshang Black breed, approximately aged between 3 and 4 years, were grouped (n = 10 goats/group) into low (D-l) and high (D-h) litter groups according to their litter size (the lower group has ≤ 2 kids/litter and the high group has ≧ 3 kids/litter, respectively). All goats were sacrificed, and collected ruminal fluid samples were subjected to 16S rRNA sequencing and LC–MS/MC Analysis for ruminal microbiome and metabolomic profiling respectively. According to PCoA analysis, the ruminal microbiota was not significantly changed by the litter sizes among the groups. The Firmicutes and Bacteroidetes were the most dominant phyla, with an abundance of 55.34% and 39.62%, respectively. However, Ruminococcaceae_UCG-009, Sediminispirochaeta, and Paraprevotella were significantly increased in the D-h group, whereas Ruminococcaceae_UCG-010 and Howardella were found to be significantly decreased in the D-l group. The metabolic profiling analysis revealed that litter size impacts metabolites as 29 and 50 metabolites in positive and negative ionic modes respectively had significant differences in their regulation. From them, 16 and 24 metabolites of the D-h group were significantly down-regulated in the positive ionic mode, while 26 metabolites were up-regulated in the negative ionic mode for the same group. The most vibrant identified metabolites, including methyl linoleate, acetylursolic acid, O-desmethyl venlafaxine glucuronide, melanostatin, and arginyl-hydroxyproline, are involved in multiple biochemical processes relevant to rumen roles. The identified differential metabolites were significantly enriched in 12 different pathways including protein digestion and absorption, glycerophospholipid metabolism, regulation of lipolysis in adipocytes, and the mTOR signaling pathway. Spearman's correlation coefficient analysis indicated that metabolites and microbial communities were tightly correlated and had significant differences between the D-l and D-h groups. Based on the results, the present study provides novel insights into the regulation mechanisms of the rumen microbiota and metabolomic profiles leading to different fertility in goats, which can give breeders some enlightenments to further improve the fertility of Yunshang Black goats. [ABSTRACT FROM AUTHOR]

This study used next-generation sequencing to assess the impact of combined capric acid (C10) and lauric acid (C12) on the ruminal bacterial composition. Eight Holstein cows were randomly assigned to two groups using a cross-over design. The cows were fed two silage-based diets with the addition of either 100 g of stearic acid per cow per day (control), or 50 g of capric acid and 50 g of lauric acid per cow per day (C10 + C12). On day 18, 250 mL of rumen fluid was collected from each cow, and DNA was isolated, amplified, and sequenced. Treatment did not alter bacterial diversity indices, the relative abundance of archaea, nor the fiber-degrading microorganisms, except for a decrease in Fibrobacter (from 2.9% to 0.7%; p = 0.04). The relative abundance of Prevotellaceae decreased (from 39.9% to 29.6%; p = 0.009), which is notable because some members help to efficiently utilize ammonia by releasing it slowly into the rumen. Furthermore, the relative abundance of Clostridia increased (from 28.4% to 41.5%; p = 0.008), which may have aided the increased ammonia–nitrogen levels in the rumen, as this class contains hyperammonia-producing members. Our study reveals alterations in bacterial abundances with implications for rumen ammonia levels, offering insights into potential strategies for modulating rumen fermentation processes and methane production in ruminant livestock. [ABSTRACT FROM AUTHOR]

Fungal probiotics have the potential as feed additives, but less has been explored in ruminant feed up to date. This study aimed to determine the effect of compound probiotics (CPs) with Aspergillus oryzae 1, Aspergillus oryzae 2 and Candida utilis on Hu sheep’s growth performance, rumen fermentation and microbiota. A total of 120 male Hu sheep, aged 2 months and with the body weight of 16.95 ± 0.65 kg were divided into 4 groups. Each group consisted of 5 replicates, with 6 sheep per replicate. Group A was the control group fed with the basal diet. Group B, C and D was supplemented with the basal diet by adding 400, 800 and 1,200 grams per ton (g/t) CPs, respectively. The feeding trial lasted for 60 days after a 10-day adaptation period. The results showed that the average daily gain (ADG) of sheep in the CPs groups were significantly higher, the feed/gain were significantly lower than those in group A in the later stage and the overall period. The addition of CPs increased the economic benefit. The levels of CD4+ and the CD4+/CD8+ ratio in the CPs groups were higher than those in Group A. The levels of GSH, IgG, IL-2, IL-6, and IFN-γ in group C were significantly elevated compared with group A. Group B showed a significant increase in rumen NH3-N and cellulase activity. There was no difference in VFAs content between group A and group B, however, with the increasing addition of CPs, the butyric acid and isobutyric acid content tended to decrease. The rumen microbiota analysis indicated that the CPs addition increased the Firmicutes and Proteobacteria abundances, decreased the Bacteroidetes abundance. The correlation analysis showed that Prevotella was negatively correlated with ADG, and the addition of 400 CPs in group B reduced Prevotella’s relative abundance, indicating CPs increased sheep growth by decreasing Prevotella abundance. The CPs addition reduced caspase-3, NF-κB and TNF-α expression in liver, jejunum and rumen tissues. In conclusion, the addition of CPs increased the sheep production performance, reduced inflammation, improved rumen and intestinal health. Considering the above points and economic benefits, the optimal addition of CPs as an additive for Hu sheep is 800 g/t.

In this study, organic acids were demonstrated as a promising carbon source for bisabolene production by the non-conventional yeast, Rhodosporidium toruloides, at microscale with a maximum titre of 1055 ± 7 mg/L. A 125-fold scale-up of the optimal process, enhanced bisabolene titres 2.5-fold to 2606 mg/L. Implementation of a pH controlled organic acid feeding strategy at this scale lead to a further threefold improvement in bisabolene titre to 7758 mg/L, the highest reported microbial titre. Finally, a proof-of-concept sequential bioreactor approach was investigated. Firstly, the cellulolytic bacterium Ruminococcus flavefaciens was employed to ferment cellulose, yielding 4.2 g/L of organic acids. R. toruloides was subsequently cultivated in the resulting supernatant, producing 318 ± 22 mg/L of bisabolene. This highlights the feasibility of a sequential bioprocess for the bioconversion of cellulose, into biojet fuel candidates. Future work will focus on enhancing organic acid yields and the use of real lignocellulosic feedstocks to further enhance bisabolene production.

Numerous studies have demonstrated that Broussonetia papyrifera is an unconventional feed resource with significant developmental potential. This research aimed to explore the effects of Broussonetia papyrifera silage on the growth performance, blood parameters, immunity, antioxidation, cytokine levels, and rumen bacterial composition of Kazakh lamb. Forty healthy male Kazakh lambs, aged 5 months and weighing 30.12 ± 1.14 kg, were randomly divided into control and experimental groups, each consisting of four replicates (five lambs per replicate). The control group was fed a basal diet, while the experimental group received a diet supplemented with 20% Broussonetia papyrifera silage (dry matter basis). Following a 10-day pre-feeding period, a 60-day formal experiment was conducted. The results indicated no significant difference in growth performance between the experimental and control groups. However, compared to the control group, the use of Broussonetia papyrifera silage significantly reduced (p < 0.05) neutrophil, lymphocyte, and eosinophil counts, as well as creatinine levels in the blood. Furthermore, Broussonetia papyrifera silage (p < 0.01) enhanced total serum antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, immunoglobulin A, immunoglobulin M, immunoglobulin G, interleukin-2, interleukin-6, and interleukin-8, and decreased malondialdehyde and interleukin-4 levels. Additionally, the use of Broussonetia papyrifera silage increased the diversity and richness of the rumen bacterial community, notably enhancing the relative abundance of Firmicutes such as Rikenellaceae_RC9_gut_group and Christensenellaceae_R-7_group. In conclusion, feeding Kazakh lamb with Broussonetia papyrifera silage (20% DM) did not adversely affect their growth performance but improved their immunity and antioxidant capacity and enhanced the relative abundance of beneficial bacteria in the rumen, thereby promoting animal health.

ABSTRACT: Some cellulolytic bacteria cannot transport branched-chain AA (BCAA) and do not express complete synthesis pathways, thus depending on cross-feeding for branched-chain volatile fatty acid (BCVFA) precursors for membrane lipids or for reductive carboxylation to BCAA. Our objective was to assess BCVFA uptake for BCAA synthesis in continuous cultures administered high forage (HF) and low forage (LF) diets without or with corn oil (CO). We hypothesized that BCVFA would be used for BCAA synthesis more in the HF than in LF diets. To help overcome bacterial inhibition by polyunsaturated fatty acids in CO, BCVFA usage for bacterial BCAA synthesis was hypothesized to decrease when CO was added to HF diets. The study was an incomplete block design with 8 dual-flow fermenters used in 4 periods with 8 treatments (n = 4) arranged as a 2 × 2 × 2 factorial. The factors were: HF or LF (67 or 33% forage, 33:67 alfalfa:orchardgrass pellets), without or with supplemental CO (3% of dry matter), and without or with 2.15 mmol/d (5 mg/d 13C) each of isovalerate, isobutyrate, and 2-methylbutyrate for one combined BCVFA treatment. The flow of bacterial BCAA increased by 10.7% by supplementing BCVFA and 9.14% with LF versus HF; similarly, dosing BCVFA versus without BCVFA increased BCAA by 1.98% in total bacterial AA, whereas LF increased BCAA by 1.92% versus HF. Additionally, BCVFA supplementation increased bacterial AA flow by 16.6% when supplemented in HF − CO and 12.4% in LF + CO diets, but not in the HF + CO (−1.5%) or LF − CO (+6.7%) diets (Diet × CO × BCVFA interaction). The recovery of 13C in bacterial AA flow was 31% lower with LF than with HF. Of the total 13C recovered in bacteria, 13.8, 17.3, and 30.2% were recovered in Val, Ile, and Leu, respectively; negligible 13C was recovered in other AA. When fermenters were dosed with BCVFA, nonbacterial and total effluent flows of AA, particularly of alanine and proline, suggest decreased peptidolysis. Increased ruminal outflow of bacterial AA, especially BCAA, but also nonbacterial AA could potentially support postabsorptive responses from BCVFA supplementation to dairy cattle.

Dietary crude protein concentration oscillation can improve the nitrogen utilization efficiency of ruminants. However, little is known about the dynamic changes in microbiota and fermentation in the rumen of calves during the oscillation period. In this study, six calves were fed an oscillating diet at 2-day intervals, including a high-protein diet (HP) and a low-protein diet (LP). The rumen fermentation parameters, plasma urea-N concentration, and rumen bacterial diversity were characterized throughout the oscillation period. The concentrations of volatile fatty acids, NH3-N, and plasma urea-N in rumen changed significantly with an oscillating diet. The abundance of Prevotella_1, Selenomonadales, Succiniclasticum, Clostridiales, Ruminococcaceae, Lachnospiraceae, and Rikenellaceae_RC9_gut_group showed significant changes with diet. Prevotella_1 was positively correlated, and Lachnospiraceae_AC2044_group and Saccharofermentans were negatively correlated with NH3-N. The abundance of Amino Acid Metabolism, Metabolism of Other Amino Acids, and Glycan Biosynthesis and Metabolism pathways, annotated by bacterial functional genes, decreased when the diet changed from HP to LP. The abundance of the Carbohydrate Metabolism pathway increased after the two dietary changes. In conclusion, the plasma urea-N concentration was not as sensitive and quick to adapt to diet changes as the rumen fermentation parameters. Rumen bacteria were responsible for increasing the nitrogen utilization efficiency of calves fed an oscillating diet.

The aim of the study was to assess the impact of potential probiotic strains of lactic acid bacteria (LAB), Apilactobacillus kunkeei EIR/BG-1 isolated from the gut of honeybees, and Enterococcus hirae EIR/CM-2 isolated from cow's milk, on rumen fermentation parameters and microbial population in a highconcentrate diet using a rumen simulation technique (Rusitec). The experiment consisted of 7 days of adaptation and 7 days of data collection. The dietary treatments were as follows: no additives (control), addition of 1 ml/fermenter (108 CFU) of Al. kunkeei EIR/BG-1, and addition of 1 ml/fermenter (108 CFU) of E. hirae EIR/CM-2. Alongside rumen fermentation characteristics, rumen microbial composition was investigated using real-time PCR. Supplementation with LAB strains did not affect ruminal pH, production of methane, and total and individual short chain fatty acids, ammonia-N concentration, dry matter digestibility and total protozoa. However, the abundance of Ruminococcus flavefaciens increased in the Al. kunkeei EIR/BG-1 treatment (P < 0.05). Moreover, the size of R. flavefaciens population in E. hirae EIR/CM-2 was comparable to that in the Al. kunkeei EIR/BG-1 treatment. Selenomonas ruminantium was more abundant in the Al. kunkeei EIR/BG-1 treatment compared to the E. hirae EIR/CM-2 treatment (P < 0.05). The abundance of Streptococcus bovis and Megasphaera elsdenii decreased with both Al. kunkeei EIR/BG-1 and E. hirae EIR/CM-2 supplementations (P < 0.05). In conclusion, Al. kunkeei EIR/BG-1 and E. hirae EIR/CM-2 exhibited beneficial effects on some members of the rumen microbial population, although these effects did not manifest in significant alterations in ruminal fermentation. Further research is required to clarify the probiotic potentials of these LAB strains as feed additives for ruminant rations. [ABSTRACT FROM AUTHOR]

This study investigated the effects of replacing hybrid giant napiers with sugarcane bagasse and fermented sugarcane bagasse on the growth performance, apparent nutrient digestibility, rumen fermentation characteristics, and rumen microorganisms of Simmental crossbred cattle. Twenty-one Simmental crossbred cattle with similar initial body weight (363.42 ± 8.67 kg) were randomly divided into three groups: Group CON (20% hybrid giant napier +45% distillers grains +35% concentrate mixture), Group SB (20% sugarcane bagasse +45% distillers grains +35% concentrate mixture), and Group FSB (20% fermented sugarcane bagasse +45% distillers grains +35% concentrate mixture). The average daily weight gain in the SB group was lower than in the CON group, no significant difference was found between the CON and FSB groups. The feed conversion ratio of the CON and FSB groups was lower compared to the SB group. The apparent digestibility of neutral detergent fiber and acid detergent fiber in the SB group was lower than in the CON group, no significant difference was found between the CON and FSB groups. The levels of NH3-N, microbial protein, acetate, propionate, butyrate, isobutyrate, and total volatile fatty acids were higher in the CON and FSB groups than in the SB group, no significant difference was found between the CON and FSB groups. The relative abundances of Christensenellaceae_R-7_group, Rikenellaceae_RC9_gut_group, Prevotellaceae_UCG-003, Saccharofermentans, and Eubacteriumcoprostanoligenes_group were lower in the CON and FSB groups compared to the SB group. The relative abundance of Succiniclasticum was highest in the FSB group, followed by the CON group and then the SB group. Correlation analysis showed that the relative abundance of Succiniclasticum was positively correlated with propionate and NH3-N content, while the relative abundance of Rikenellaceae_RC9_gut_group was inversely correlated with NH3-N content. Gene function prediction indicated that fermented sugarcane bagasse promoted rumen microbial amino acid metabolism. In conclusion, replacing hybrid giant napiers with 20% sugarcane bagasse negatively affected the growth performance of Simmental crossbred cattle, while the addition of 20% fermented sugarcane bagasse had no adverse effects on growth performance and rumen fermentation characteristics, and did not alter the abundance of the rumen core flora in Simmental crossbred cattle. [ABSTRACT FROM AUTHOR]

This study used next-generation sequencing to assess the impact of combined capric acid (C10) and lauric acid (C12) on the ruminal bacterial composition. Eight Holstein cows were randomly assigned to two groups using a cross-over design. The cows were fed two silage-based diets with the addition of either 100 g of stearic acid per cow per day (control), or 50 g of capric acid and 50 g of lauric acid per cow per day (C10 + C12). On day 18, 250 mL of rumen fluid was collected from each cow, and DNA was isolated, amplified, and sequenced. Treatment did not alter bacterial diversity indices, the relative abundance of archaea, nor the fiber-degrading microorganisms, except for a decrease in Fibrobacter (from 2.9% to 0.7%; p = 0.04). The relative abundance of Prevotellaceae decreased (from 39.9% to 29.6%; p = 0.009), which is notable because some members help to efficiently utilize ammonia by releasing it slowly into the rumen. Furthermore, the relative abundance of Clostridia increased (from 28.4% to 41.5%; p = 0.008), which may have aided the increased ammonia–nitrogen levels in the rumen, as this class contains hyperammonia-producing members. Our study reveals alterations in bacterial abundances with implications for rumen ammonia levels, offering insights into potential strategies for modulating rumen fermentation processes and methane production in ruminant livestock.

ObjectiveThe objective of this study was to compare the effects of Leymus chinensis hay and alfalfa hay as the roughage on the rumen bacterial and the meat metabolomics in lambs.MethodsFourteen male lambs were randomly assigned to two dietary treatments (one group was fed with concentrate and Leymus chinensis hay; another was fed with concentrate and alfalfa hay) with seven replicates per treatment. The feeding experiment lasted for 60 days. Lambs were slaughtered at the end of the feeding experiment. Growth performance, carcass performance, and weights of various viscera were determined. The longissimus dorsi and rumen contents were collected for untargeted metabolomics and 16S rDNA amplicon sequencing analysis, respectively.ResultsThe lambs fed with alfalfa hay showed a significantly increased in average daily gain, carcass weight, dressing percentage, loin-eye area, and kidney weight. Feeding Leymus chinensis hay and alfalfa hay diets resulted in different meat metabolite deposition and rumen bacterial communities in the lambs. The relative abundance of phyla Fibrobacteres, Bacteroidetes, and Spirochaetes were greater in the Leymus Chinensis hay group, while, the relative abundance of Firmicutes, Proteobacteria, Fusobacteria, and Verrucomicrobia were greater in the alfalfa hay group. Based on untargeted metabolomics, the main altered metabolic pathways included alanine, aspartate and glutamate metabolism, D-glutamine and D-glutamate metabolism, phenylalanine metabolism, nitrogen metabolism, and tyrosine metabolism. Several bacteria genera including BF31, Alistipes, Faecalibacterium, Eggerthella, and Anaeroplasma were significantly correlated with growth performance and meat metabolites.ConclusionAlfalfa hay improved growth performance and carcass characteristics in lambs. Leymus chinensis hay and alfalfa hay caused different meat metabolite deposition by modifying the rumen bacterial community. These findings will be beneficial to future forage utilization for sheep growth, carcass performance, and meat quality improvement.

Introduction: Milk fat is the most variable nutrient in milk, and recent studies have shown that rumen bacteria are closely related to milk fat. However, there is limited research on the relationship between rumen bacteria and milk fatty. Fatty acids (FAs) are an important component of milk fat and are associated with various potential benefits and risks to human health. Methods: In this experiment, forty-five healthy Holstein dairy cows with alike physiological and productive conditions were selected from medium-sized dairy farms and raised under the same feeding and management conditions. The experimental period was two weeks. During the experiment, raw milk and rumen fluid were collected, and milk components were determined. In this study, 8 high milk fat percentage (HF) dairy cows and 8 low milk fat percentage (LF) dairy cows were selected for analysis. Results: Results showed that the milk fat percentage in HF group was significantly greater than that of the dairy cows in the LF group. 16S rRNA gene sequencing showed that the rumen bacterial abundance of HF dairy cows was significantly higher than that in LF dairy cows; at the genus level, the bacterial abundances of Prevotellaceae_UCG-001, Candidatus_Saccharimonas, Prevotellaceae_UCG-003, Ruminococcus_1, Lachnospiraceae_XPB1014_group, Lachnospiraceae_AC2044_ group, probable_genus_10 and U29-B03 in HF group were significantly higher than those in the LF group. Spearman rank correlation analysis indicated that milk fat percentage was positively related to Prevotellaceae_UCG-001, Candidatus_ Saccharimonas, Prevotellaceae_UCG-003, Ruminococcus_1, Lachnospiraceae_ XPB1014_group, Lachnospiraceae_AC2044_group, probable_genus_10 and U29-B03. Furthermore, Prevotellaceae_UCG-001 was positively related to C14:0 iso, C15:0 iso, C18:0, Ruminococcus_1 with C18:1 t9, Lachnospiraceae_AC2044_ group with C18:1 t9 and C18:1 t11, U29-B03 with C15:0 iso. Discussion: To sum up, rumen bacteria in dairy cows are related to the variation of milk fat, and some rumen bacteria have potential effects on the deposition of certain fatty acids in raw milk. [ABSTRACT FROM AUTHOR]

Background: High-producing dairy cows face varying degrees ofmetabolic stress and challenges during the late perinatal period, resulting in ruminal bacteria abundance and their fermentative ability occurring as a series of changes. However, the dynamic changes are still not clear. Aims/methods: Ten healthy, high-producing Holstein dairy cows with similar body conditions and the same parity were selected, and ruminal fluid from the dairy cows at postpartum0, 7, 14, and 21 d was collected beforemorning feeding. 16S rRNA high-throughput sequencing, GC-MS/MS targeted metabolomics, and UPLC-MS/MS untargeted metabolomics were applied in the study to investigate the dynamic changes within 21 d postpartum. Results: The results displayed that the structures of ruminal bacteria were significantly altered from 0 to 7 d postpartum (R = 0.486, P = 0.002), reflecting the significantly declining abundances of Euryarchaeota and Chloroflexi phyla and Christensenellaceae, Methanobrevibacter, and Flexilinea genera (P < 0.05) and the obviously ascending abundances of Ruminococcaceae, Moryella, Pseudobutyrivibrio, and Prevotellaceae genera at 7 d postpartum (P < 0.05). The structures of ruminal bacteria also varied significantly from 7 to 14 d postpartum (R = 0.125, P = 0.022), reflecting the reducing abundances of Christensenellaceae, Ruminococcaceae, and Moryella genera (P < 0.05), and the elevating abundances of Sharpea and Olsenella genera at 14 d postpartum (P < 0.05). The metabolic profiles of ruminal SCFAs were obviously varied from 0 to 7 d postpartum, resulting in higher levels of propionic acid, butyric acid, and valeric acid at 7 d postpartum (P < 0.05); the metabolic profiles of other ruminal metabolites were significantly shifted from 0 to 7 d postpartum, with 27 significantly elevated metabolites and 35 apparently reduced metabolites (P < 0.05). The correlation analysis indicated that propionic acid was positively correlated with Prevotellaceae and Ruminococcaceae (P < 0.05), negatively correlated with Methanobrevibacter (P < 0.01); butyric acid was positively associated with Prevotellaceae, Ruminococcaceae, and Pseudobutyrivibrio (P < 0.05), negatively associated with Christensenellaceae (P < 0.01); valeric acid was positively linked with Prevotellaceae and Ruminococcaceae (P < 0.05); pyridoxal was positively correlated with Flexilinea and Methanobrevibacter (P < 0.05) and negatively correlated with Ruminococcaceae (P < 0.01); tyramine was negatively linked with Ruminococcaceae (P < 0.01). Conclusion: The findings contribute to the decision of nutritional management and prevention ofmetabolic diseases in high-producing dairy cows during the late perinatal period. [ABSTRACT FROM AUTHOR]

Hydrogen (H2) is the primary electron donor for methane formation in ruminants, but the H2-producing organisms involved are largely uncharacterized. This work integrated studies of microbial physiology and genomics to characterize rumen bacterial isolate NK3A20 of the family Lachnospiraceae. Isolate NK3A20 was the first recognized isolate of the NK3A20 group, which is among the ten most abundant bacterial genera in 16S rRNA gene surveys of rumen microbiota. NK3A20 produced acetate, butyrate, H2, and formate from glucose. The end product ratios varied when grown with different substrates and at different H2 partial pressures. NK3A20 produced butyrate as a major product using glucose or under high H2 partial pressures and switched to mainly acetate in the presence of galacturonic acid (an oxidized sugar) or in coculture with a methanogen. Growth with galacturonic acid was faster at elevated H2 concentrations, while elevated H2 slowed growth with glucose. Genome analyses revealed the presence of multiple hydrogenases including a membrane-bound Ech hydrogenase, an electron bifurcating butyryl-CoA dehydrogenase (Bcd-Etf), and an Rnf complex that may be involved in modulating the observed metabolic pathway changes, providing insight into H2 formation in the rumen. [ABSTRACT FROM AUTHOR]

Background. Previous studies have discussed the effects of grazing and house feeding on yaks during the cold season when forage is in short supply, but there is limited information on the effects of these feeding strategies on Jersey cows introduced to the Tibetan Plateau. The objective of this study was to use genomics and metabolomics analyses to examine changes in rumen microbiology and organism metabolism of Jersey cows with different feeding strategies. Methods. We selected 12 Jersey cows with similar body conditions and kept them for 60 days under grazing (n D 6) and house-feeding (n D 6) conditions. At the end of the experiment, samples of rumen fluid and serum were collected from Jersey cows that had been fed using different feeding strategies. The samples were analyzed for rumen fermentation parameters, rumen bacterial communities, serum antioxidant and immunological indices, and serum metabolomics. The results of the study were examined to find appropriate feeding strategies for Jersey cows during the cold season on the Tibetan plateau. Results. The results of rumen fermentation parameters showed that concentrations of acetic acid, propionic acid, and ammonia nitrogen in the house-feeding group (Group B) were significantly higher than in the grazing group (Group G) (P <0:05). In terms of the rumen bacterial community 16S rRNA gene, the Chao1 index was significantly higher in Group G than in Group B (P D0:038), while observed species, Shannon and Simpson indices were not significantly different from the above-mentioned groups (P >0:05). Beta diversity analysis revealed no significant differences in the composition of the rumen microbiota between the two groups. Analysis of serum antioxidant and immune indices showed no significant differences in total antioxidant capacity between Group G and Group B (P > 0:05), while IL-6, Ig-M, and TNF-α were significantly higher in Group G than in Group B (P <0:05). LC≥MS metabolomics analysis of serum showed that a total of 149 major serum differential metabolites were found in Group G and Group B. The differential metabolites were enriched in the metabolic pathways of biosynthesis of amino acids, protein digestion and absorption, ABC transporters, aminoacyl-tRNA biosynthesis, mineral absorption, and biosynthesis of unsaturated fatty acids. These data suggest that the house-feeding strategy is more beneficial to improve the physiological state of Jersey cows on the Tibetan Plateau during the cold season when forages are in short supply. [ABSTRACT FROM AUTHOR]

Aim: The aim of this study was to investigate the in vitro dose-dependent effects of sigla storax (Styrax liquidus) on rumen microbiota and rumen microbial fermentation in comparison to monensin as a positive control. Methods and Results: This study was carried out using a rumen simulation model (Rusitec). Treatments consisted of no additive (control), 10 mg l−1 of monensin sodium salt, 100 mg l−1 (Low-Sigla), and 500 mg l−1 (High-Sigla) of sigla storax (n = 6/treatment). In addition to rumen fermentation characteristics, rumen microbial composition was investigated using 16S rRNA sequencing. The methane variables and the acetate to propionate ratio decreased in the both High-Sigla and monensin groups (P < 0.05). High-Sigla had no effect on ammonia, total SCFA and nutrition degradation, while monensin decreased these parameters (P < 0.05). Unlike monensin, the sigla storax treatments did not affect the alpha or beta diversity indexes of the microbiota. The relative abundance of Methanomethylophilaceae and Ruminococcaceae decreased with High-Sigla and monensin (P < 0.05), and Atopobiaceae and Eggerthellaceae decreased with the both doses of sigla storax as well as monensin treatments (P < 0.05). Syntrophococcus, DNF00809, and Kandleria were among the genera that most decreased with High-Sigla and monensin (Q < 0.07) and were strongly positively correlated with methane production (r = 0.52–0.56). Conclusions: The high dose of sigla storax (500 mg l−1) decreased methane in the rumen ecosystem without adverse effects on nutrient degradation and SCFA production, and without dramatically impacting the microbial composition. Sigla storax might be a novel feed additive to mitigate methane in cattle. [ABSTRACT FROM AUTHOR]

ABSTRACT: This research aimed to investigate the effects of replacing soybean meal with high-oil pumpkin seed cake (HOPSC) on ruminal fermentation, lactation performance, milk fatty acid, and ruminal bacterial community in Chinese dairy cows. Six multiparous Chinese Holstein cows at 105.50 ± 5.24 d in milk (mean ± standard deviation) and 36.63 ± 0.74 kg/d of milk yield were randomly allocated, in a 3 × 3 Latin square design, to 3 dietary treatments in which HOPSC replaced soybean meal. Group 1 was the basal diet with no HOPSC (0HOPSC); group 2 was a 50% replacement of soybean meal with HOPSC and dried distillers grains with solubles (DDGS; 50HOPSC), and group 3 was a 100% replacement of soybean meal with HOPSC and DDGS (100HOPSC). We found no difference in the quantity of milk produced or milk composition among the 3 treatment groups. Feed efficiency tended to increase linearly as more HOPSC was consumed. In addition, rumen fermentation was not influenced when soybean meal was replaced with HOPSC and DDGS; the relative abundance of ruminal bacteria at the phylum and genus levels was altered. We also observed that as the level of HOPSC supplementation increased, the relative abundance of Firmicutes and Tenericutes linearly increased, whereas that of Bacteroidetes decreased. However, with increasing HOPSC supplementation, the relative abundance of Ruminococcus decreased linearly at the genus level in the rumen, and the relative abundance of Prevotella showed a linear downward tendency. Changes in dietary composition and rumen bacteria had no significant effect on the fatty acid composition of milk. In conclusion, our results indicated that replacing soybean meal with a combination of HOPSC and DDGS can meet the nutritional needs of high-yielding dairy cows without adversely affecting milk yield and quality; however, the composition of rumen bacteria could be modified. Further study is required to investigate the effects of long-term feeding of HOPSC on rumen fermentation and performance of dairy cows.

The effects of different inclusion rates of Poncirus trifoliata (PT) extract on in vitro ruminal fermentation parameters were investigated. Six dose levels were referred to as follows: PT0, control (without PT); PT1, (3 mg/L); PT3, (9 mg/L); PT5, (15 mg/L); PT7, (21 mg/L); PT9, (27 mg/L), each incubated with 50 mL of buffered rumen fluid and 300 mg of timothy hay as a substrate for 72 h of incubation using an in vitro batch culture system. The in vitro dry matter digestibility (IVDMD) was significantly affected by the PT dose, except after 6 and 48 h of incubation. Total volatile fatty acids (VFAs) increased quadratically (12 h of incubation) within the range of PT1–PT7, whereas decreased at PT9 dose. Methane (CH4) production and the proportion of CH4 in the total gas produced decreased linearly after 12 h of incubation. The relative abundance of Ruminococcus albus and Ruminoccocus flavefaciens was greater at the doses PT1 than PT0, whereas that of Fibrobacter succinogenes was greater at PT0 than at PT9 after 24 h of incubation. Methanogenic archaea decreased linearly with increasing PT dose after 12 h of incubation. The abundance of ciliate-associated methanogens decreased linearly over 24 h of incubation under PT7 and PT9 treatments. These results suggest that the dose of PT can modulate ruminal fermentation to alter VFA concentration and reduce CH4 production by altering ruminal bacterial abundance.Highlights This study evaluated the dose-response effects of Poncirus trifoliata as a methane mitigation agent in ruminants. Inclusion of Poncirus trifoliate lowered in vitro dry matter digestibility and total volatile fatty acid concentration. Inclusion of Poncirus trifoliata extract reduced methane production after 12 h of incubation but did not maintained. Inclusion of Poncirus trifoliata extract reduced methanogenic archaea and ciliate-associated methanogens abundance

Introduction: The Tibetan Plateau is characterized by low temperature and hypoxia. N-carbamylglutamic acid (NCG) can increase blood oxygen saturation, and have the potential to be used to prevent the high-altitude hypoxia stress state of cows. However, its beneficial effect on the rumen microbiota of Holstein dairy cows remains unclear. Methods: Hence, the experiments 12 multiparous (parity ranged from 2 to 7) Holstein dairy cows (413.0 ± 42 kg) were randomly assigned to 2 treatments with 6 replicates in each treatment: basal diet (CON, control group) and basal diet plus 20 g/d/cow of NCG (NCG, experiment group), respectively. To study the effects of dietary NCG supplementation on rumen microbiota of Holstein dairy cows in Tibet. The experiment lasted for 45 days, with 15 days of pre-feeding and 30 days of formal trail period. Results: The results showed that ruminal NH3-N concentration in NCG group was lower (p < 0.05) than that in the CON group, while molar proportion of acetic acid and total volatile fatty acid (VFA) concentration were increased (p < 0.05) with the addition of NCG. Microbial diversity increased (p < 0.05) in NCG group, with Bacteroidetes, Firmicutes, and Patescibacteria as the most abundant phyla. The KEGG pathway analysis showed that the potential function of ruminal bacteria was mainly enriched in metabolism (carbohydrates, amino acids, lipids, energy, and nucleotides) and genetic information processing (replication, repair, and translation). Conclusion: In conclusion, NCG can improve rumen nitrogen utilization, total VFA and acetic acid production, and increase rumen microbial diversity, all of which could make the introduced Holstein dairy cows to better adapt to the harsh environment in Tibet and improve their production performance. [ABSTRACT FROM AUTHOR]

Efficient feed utilization in dairy cows is crucial for economic and environmental reasons. The rumen microbiota plays a significant role in feed efficiency, but studies utilizing microbial data to predict host phenotype are limited. In this study, 87 primiparous Nordic Red dairy cows were ranked for feed efficiency during their early lactation based on residual energy intake, and the rumen liquid microbial ecosystem was subsequently evaluated using 16S rRNA amplicon and metagenome sequencing. The study used amplicon data to build an extreme gradient boosting model, demonstrating that taxonomic microbial variation can predict efficiency (rtest = 0.55). Prediction interpreters and microbial network revealed that predictions were based on microbial consortia and the efficient animals had more of the highly interacting microbes and consortia. Rumen metagenome data was used to evaluate carbohydrate-active enzymes and metabolic pathway differences between efficiency phenotypes. The study showed that an efficient rumen had a higher abundance of glycoside hydrolases, while an inefficient rumen had more glycosyl transferases. Enrichment of metabolic pathways was observed in the inefficient group, while efficient animals emphasized bacterial environmental sensing and motility over microbial growth. The results suggest that inter-kingdom interactions should be further analyzed to understand their association with the feed efficiency of animals. [ABSTRACT FROM AUTHOR]

ObjectiveThe purpose of this study was to investigate the effects of different doses of Sophora alopecuroides (SA) on the rumen fermentation and microbial diversity of sheep.MethodsA total of 32 healthy Dumont crossbred male lambs weighing 25.73 ± 2.17 kg were randomly assigned to 4 treatment groups with 8 replicates each: a control group (CG) fed a basal diet with a concentrate-to-forage ratio of 7:3 and three experimental groups - the 0.1% group(TG1), 0.3% group (TG2), and 0.5% group (TG3), which were fed the same basal diet but supplemented with increasing doses of SA.Results(1) Increasing the SA dose led to a significant linear increase (p-< 0.05) in acetate, propionate, butyrate, and total volatile fatty acid (TVFA) concentrations in the rumen, as well as a significant quadratic effect (p-< 0.05) on the propionate concentration. In contrast, there was a significant linear decrease (p-< 0.05) in the NH3-N concentration in the rumen. (2) At the level of rumen bacterial phyla, the abundance of Bacteroidetes in the rumen increased, and that of Firmicutes decreased (p = 0.08). At the genus level, the rumen abundances of Ruminococcus and Phocaeicola of sheep in the three experimental groups were significantly higher than in the control group (p-< 0.05), and the abundances of Clostridiales and Candidatus-Hepatincola were significantly increased in the 0.1% and 0.3% groups (p < 0.05). (3) Regarding rumen anaerobic fungi, the differences between the control group and experimental groups at the phylum level and genus level were not significant (p > 0.05), but the relative abundances of Neocallimastigomycota and Piromyces in the 0.1% group were significantly higher than that in the control group.ConclusionSA addition to a high grain diet could increase the VFA concentration and pH in the sheep rumen, reduce the NH3-N concentration in the rumen and improve rumen fermentation function. Although there was no significant change in rumen bacterial or fungal diversity, SA addition increased the rumen abundances of Bacteroidetes, Ruminococcus, Phocaeicola, Clostridiales, Neocallimastigomycota and Piromyces, decreased the rumen abundance of Firmicutes, and had a positive effect on the rumen microbiota to improve sheep health.

Sweet sorghum silage (SS; Sorghum dochna ‘Dochna’) has been extensively studied in recent years as a supplementary forage-to-corn silage (CS; Zea mays L.), but there are still relatively few studies on its effects on the rumen environment of sheep. Determining the short-term impact of converting roughage from corn straws to SS compared to CS on rumen fermentation and bacterial population dynamics was the main goal of the current study. Twelve female thin-tailed Han sheep (29.8 ± 1.34 kg) were randomly divided into one of two treatments: concentrate supplemented with SS or CS, respectively. During the 15-day pretest period, concentrate was fed in two separate feedings at 0800 h and 1800 h, and ensure that the animals were all consumed within an hour of being fed. Thereafter, the animals had free access to corn straw. The feeding procedures during the pretest period were the same as during the measurement period. Rumen fluid was collected via sheep esophageal tube on the last day of adaptation phase (1–7 days) and stabilisation phase (8–30 days), respectively. The results showed that there was a similarity in the total concentration of VFA (volatile fatty acid) and the proportions of acetate, propionate, butyrate, and branched-chain VFA (P > 0.05) and microbial diversity indices (P > 0.05) between the two silage groups throughout the experimental period. The concentration of Ammonia nitrogen (P = 0.001) and proportion of valerate (P = 0.028) decreased in the CS and SS groups, respectively. The abundance and predicted function of rumen bacteria in the SS group did not differ significantly (P > 0.05) between the two measurement phases. However, the abundance of Prevotella_1 (P = 0.038) was higher in the CS group than in the SS group at 7 d. The abundances of Firmicutes (P = 0.005) and Ruminococcaceae_NK4A214_group (P = 0.002) increased, while the abundances of Bacteroidetes (P = 0.044), Proteobacteria (P = 0.046), and Prevotella_1 (P = 0.009) decreased in the CS group at 30 d. Genes related to pyruvate metabolism (P = 0.020) were significantly higher at 30 d than at 7 d, whereas purine metabolism (P = 0.007), pyrimidine metabolism (P = 0.007), and metabolic pathways (P = 0.010) were lower at 30 d in the CS group. In conclusion, this study indicated that SS maintained a steady rumen environment, while CS caused high fluctuations in bacterial abundance and predicted function for sheep.

Microbial fuel cells (MFCs) using rumen bacteria have been proposed as a power source for running devices inside cattle. In this study, we explored the key parameters of the conventional bamboo charcoal electrode in an attempt to improve the amount of electrical power generated by the microbial fuel cell. We evaluated the effects of the electrode's surface area, thickness, and rumen content on power generation and determined that only the electrode's surface area affects power generation levels. Furthermore, our observations and bacterial count on the electrode revealed that rumen bacteria concentrated on the surface of the bamboo charcoal electrode and did not penetrate the interior, explaining why only the electrode's surface area affected power generation levels. A Copper (Cu) plate and Cu paper electrodes were also used to evaluate the effect of different electrodes on measuring the rumen bacteria MFC's power potential, which had a temporarily higher maximum power point (MPP) compared to the bamboo charcoal electrode. However, the open circuit voltage and MPP decreased significantly over time due to the corrosion of the Cu electrodes. The MPP for the Cu plate electrode was 775 mW/m2 and the MPP for the Cu paper electrode was 1240 mW/m2, while the MPP for bamboo charcoal electrodes was only 18.7 mW/m2. In the future, rumen bacteria MFCs are expected to be used as the power supply of rumen sensors. [ABSTRACT FROM AUTHOR]

Cattle milk is an important food for the growing calf and humans because of its concentrated macro and micro nutrients. However, the quantity and quality of milk nutrients vary depending on several factors. This study evaluated the effect of dietary modification on rumen bacteria as well as the potential role of rumen bacteria in modifying milk production and composition profile. Using a 16S rRNA metagenomic approach, the study characterized the rumen bacterial community composition in four dairy cows and their milk production and composition. The results indicated that Bacteroidetes and Firmicutes were the dominant bacteria, totaling ∼83.7% of the rumen bacteria. An increase in concentrate proportion in diet led to an increase in the abundance of Bacteroidetes (P  ≤ 0.05) and Proteobacteria while Firmicutes and Fibrobacter decreased. Milk production and composition were highly correlated with the abundance of various rumen bacterial members. Lentispaerae (P  = 0.010), and Synergistetes (P  = 0.011) exhibited a positive and significant correlation while Tenericutes (P  = 0.009) showed a negative correlation with milk protein. Fusobacteria (P  = 0.016) showed a negative correlation with milk lactose. Similarly, several genera showed correlations with milk parameters. The correlation between microbes with milk parameters implies that the bacterial community possesses the potential to influence milk production and composition. [ABSTRACT FROM AUTHOR]

The performance of ruminants is affected mainly by the rumen bacterial community. The composition and properties of the rumen bacterial community depend largely on the diet components that are fed to the ruminant. The aim of this study was to determine the effect of grape pomace intake on the rumen bacterial community of sheep. Four different diets, two of which contained dried grape pomace (DGP), were used in this study. Rumen fluid samples from 12 wethers were used for 16S rRNA gene sequencing and subsequent bacterial identification. At the phylum level, Bacteroidetes and Firmicutes; at the family level, Prevotellaceae and Porphyromonadaeceae; and at the genera level, Prevotella and Verrucomicrobia_Subdivision5_genera_incertae_sedis were the most common regardless of the diet the animals were fed. After the addition of DGP to the diet, the relative abundance of Methanobrevibacter, Butytirivibrio, Fretibacterium, and Verrucomicrobia _Subdivision3 _genera_incertae_sedis significantly increased, whereas that of Succiniclasticum and Selenomonas significantly decreased. The upregulated pathway of methanogenesis from H2 and CO2 was supported by a significant increase in Methanobacteriaceae after the diet was supplemented with DGP. The rumen bacterial community diversity indices (Richness and Shannon) were significantly affected by diet composition as well as by the change of housing location. The addition of DGP into the wethers' diet increased the richness of the rumen bacterial community, which is good for maintaining rumen bacterial homeostasis. No adverse effect of the addition of DPG on the rumen bacterial community was noted. [ABSTRACT FROM AUTHOR]

Total milk solid (TMS) content directly reflects the quality of milk. Rumen bacteria ferment dietary components, the process of which generates the precursors for the synthesis of milk solid, therefore, the variation in rumen bacterial community could be associated with milk solid in dairy cows. In this study, 45 healthy mid-lactation Holstein dairy cows with the similar body weight, lactation stage, and milk yield were initially used for the selection of 10 cows with high TMS (HS) and 10 cows with low TMS (LS). All those animals were under the same feeding management, and the individual milk yield was recorded for 14 consecutive days before milk and rumen fluid were sampled. Rumen fluid was used to determine bacterial community by 16S rRNA gene sequencing technique. The HS cows had significantly greater feed intake and milk TMS, fat, protein content than LS cows (P < 0.05). Among the volatile fatty acids (VFA), propionic acid and valeric acid concentrations were significantly greater in HS cows than those in LS cows (P < 0.05). There was no significant difference in the concentrations of acetate, butyrate, isobutyrate, valerate, and the total VFA (P > 0.05), nor was the acetate-to-propionate ratio, pH value, ammonia nitrogen and microbial crude protein concentrations (P > 0.05). Significant differences in the relative abundances of some bacterial genera were found between HS and LS cows. Spearman’s rank correlation analysis revealed that TMS content was correlated positively with the abundances of Ruminococcaceae UCG-014, Ruminococcaceae NK4A214 group, Prevotellaceae UCG-001, Butyrivibrio 2, Prevotellaceae UCG-003, Candidatus Saccharimonas, Ruminococcus 2, Lachnospiraceae XPB1014 group, probable genus 10, Eubacterium ventriosum group, but negatively correlated with Pyramidobacte. In addition, Ruminococcaceae UCG-014, Ruminococcus 2, Ruminococcaceae UCG001, probable genus 10 and Eubacterium ventriosum group might boost the total VFA production in the rumen. In conclusion, the dry matter intake of dairy cows and some special bacteria in rumen were significantly associated with TMS content, which suggests the potential function of rumen bacteria contributing to TMS content in dairy cows.

ABSTRACT: Forty-eight newborn Holstein dairy calves [40 ± 3.4 (SD) of kg of body weight (BW); 24 females and 24 males] were used in a completely randomized design to investigate the effects of a mixture of phytobiotic-rich herbal extract (Immunofin, IMPE) incorporated into milk on performance, ruminal fermentation, bacterial population, and serum biochemical metabolites during the preweaning period. Calves had free access to calf starter and clean water from d 6 until weaning. The treatments were the control (CON; without additive) and IMPE at 4, 8, and 12 mL/d. The treatments had no significant effect on total dry matter intake, weight gain, and BW at weaning. The incidence of diarrhea was lower in calves fed 8 mL of IMPE/d compared with CON. At weaning, body measurements (except for front leg circumference) were not affected by IMPE treatment. Relative to the CON group, front leg circumference was significantly decreased by IMPE supplementation. Serum IgG concentration was not significantly increased by IMPE supplementation compared with the CON group. Triglyceride concentration decreased in calves receiving 4, 8, and 12 mL/d of IMPE compared with the CON groups. In contrast to the CON group, serum albumin and total serum protein concentrations increased with IMPE supplementation. Calves receiving 4 mL/d of IMPE had a greater abundance of total bacteria, Ruminococcus albus, Ruminococcus flavefaciens, and Fibrobacter succinogenes compared with the other treatments. Molar proportions of acetate increased in calves fed IMPE (at 12 mL/d) compared with calves fed CON. Ruminal N-NH3 concentrations decreased linearly with the increase in IMPE supplementation. The results of the present study suggest that the addition of IMPE to milk may improve some health and immunity conditions, blood metabolite concentrations, and increase the abundance of some cellulolytic bacteria in the rumen of Holstein dairy calves. The use of IMPE may be an alternative to feeding antibiotics at subtherapeutic concentrations to improve calf health and immunity status.

The objective of this study was to investigate the effect of waste milk with antibiotic residue on rumen fermentation and rumen bacterial composition of dairy calves during pre-weaned and post-weaned periods. A total of 24 Holstein male calves (43.4 ± 0.93 kg body weight, mean ± standard error) were allocated into four blocks based on birth date. Dairy calves were supplied 100% milk replacer (MR, n = 8), 50% milk replacer mixed with 50% waste milk (MM, n = 8), or 100% waste milk (WM, n = 8). Ruminal samples were collected at 49 and 63 days of age and then subjected to determinations of pH value, volatile fatty acids (VFA), ammonia nitrogen (NH3-N) and 16S rRNA gene amplicon sequencing. The results showed that feeding WM had no effect on the pH value, the concentrations of VFA (acetic acid, propionic acid, butyric acid, isovaleric acid, valeric acid), and NH3-N in dairy calves compared to feeding MR. However, from 49 to 63 days of age, the pH value (p < 0.001) was significantly increased, while the levels of total VFA (p = 0.004), acetic acid (p = 0.01), propionic acid (p = 0.003) and valeric acid (p < 0.001) were significantly decreased. For rumen microorganisms, there was no differences in bacterial diversity among the treatments. But the relative abundance of Veillonellaceae was significantly lower (p = 0.05) in the calves fed WM than that from MR group at 49 days of age; however, no difference was detected at 63 days of age. Feeding WM to calves tended to reduce family Veillonellaceae and genus Olsenella in the rumen at 49 days of age (p = 0.049). Analysis of temporal changes in rumen bacteria based on alpha-diversity and beta-diversity as well as the microbial relative abundances did not exhibit any difference. In addition, relative abundances of Clostridia_UCG-014, Prevotella, Syntrophococcus, Eubacterium_nodatum_group, Pseudoramibacter and Solobacterium were correlated with rumen pH value and the concentrations of TVFA, propionic acid, isovaleric acid, valeric acid and NH3-N. In conclusion, compare to MR, calves supplied with WM had little changes on the rumen pH value, NH3-N or VFAs contents. Additionally, limited effects could be found on rumen microbiota in the calves fed WM. However, further studies needed to explore if there exist any long-term effects of early-life rumen microbiota modulation on dairy cows. [ABSTRACT FROM AUTHOR]

This study investigated the effects of active dry yeast (ADY) and yeast culture (YC) supplementation on rumen bacteria and metabolites in finishing bulls fed high-concentrate diets using the full-length 16S rDNA gene sequencing and liquid chromatography-mass spectrometry. Supplementation with ADY improved the alpha diversity and relative abundance of rumen bacteria, while YC only affected relative abundance of rumen bacteria at the genus level. Sixty-three differential metabolites were identified in rumen fluid after ADY supplementation, and 17 after YC. PICRUSt2 functional prediction showed that ADY supplementation improved the capacity of amino acid metabolism, lipid metabolism, carbohydrate metabolism, metabolism of terpenoids and polyketides, and energy metabolism in rumen bacteria (all P < 0.05). Correlation analysis showed that the rumen differential metabolites following ADY supplementation were mainly related to Oligosphaera, Verruc, Mycoplasma, and Anaeroplasma. Supplementation with ADY was more effective than YC in remodeling the rumen bacterial flora structure and metabolite composition under high-concentrate diets. [ABSTRACT FROM AUTHOR]

Simple Summary: Protein is a crucial nutrient for improving performance and fiber utilization by grazing ruminants. The use of non-protein sources can be used to improve the intake and digestibility of poor-quality forages, since rumen microbes can use NPN to synthesize amino acids. Urea supplementation has become common for cattle. However, urea shows a rapid hydrolysis in the rumen, which can lead to excess ammonia and can impair animal performance and risk intoxication. Supplementation with post-ruminal available urea can prevent ammonia accumulation and can be an alternative source of nitrogen for grazing cattle. The objective of this study was to evaluate the effects of post-ruminal urea supplementation in different types of supplement formulation for Nellore bulls in the growing phase during the seasonal period. Performance and rumen environmental parameters were also evaluated. Our results emphasize that replacing soybean meal as a source of true protein with non-protein nitrogen resulted in reduced performance, and that conventional urea and post-ruminal urea have similar effects on animal performance and rumen metabolism. The objective was to evaluate the effects of urea with post-ruminal absorption in the supplementation of growing Nellore cattle reared on pasture during a seasonal period. For the study, two experiments were conducted. In experiment 1, rumen and blood parameters were evaluated using eight rumen-cannulated Nellore bulls with initial body weight (BW) of 763 ± 44 kg, distributed in a double Latin square 4 × 4. In experiment 2, 120 Nellore steers with initial BW of 380 ± 35 kg were used for performance evaluation, distributed in a randomized block design (blocking factor or initial BW). The evaluated treatments were 1: (TP-U) (control) = supplement with 24% crude protein (CP) containing urea as a source of non-protein nitrogen (NPN; 3%) and soybean meal, 2: (TP-PRU) = 24% CP supplement containing post-ruminal urea (PRU; 3.6%) and soybean meal; 3: (NPN-U-PRU) = 24% CP supplement containing urea + post-ruminal urea (U = 3% and PRU = 3.9%), without soybean meal; 4: (NPN-PRU) = supplement with 24% CP containing post-ruminal urea (7.5%), without soybean meal. The supplement was offered at 3 g/kg BW per animal, daily, once a day. All animals were kept on Urochloa brizantha cv. Marandu pasture. Statistical analyses were performed using the SAS PROC MIXED, and the data were evaluated by the following contrasts: C1 = TP-U/TP-PRU vs. NPN-U-PRU/NPN-PRU (Soybean meal replacement by NPN); C2 = TP-U vs. TP-PRU (conventional urea vs. post-immune urea); C3 = NPN-U-PRU vs. NPN-PRU (low and high post-ruminal urea-PRU level). The digestibility of dry matter, organic matter, and NDF was lower when soybean meal was replaced by non-protein nitrogen, also being different between the levels of post-ruminal urea used in the supplement. Ruminal pH was different when soybean meal was replaced by NPN (p = 0.003). Total concentration of short-chain fatty acids, concentrations of isobutyrate (p = 0.003), valerate (p = 0.001), and isovalerate (p = 0.001) were different, and blood urea was different when soybean meal was replaced by NPN (p = 0.006). Simpson's diversity index was higher in the rumen of animals supplemented with TP-U than in those supplemented with TP-PRU (p = 0.05). A total of 27 phyla, 234 families, and 488 genera were identified. Nitrospirota and Gemmatimonadota phyla were detected just in the rumen of steers supplemented with TP-PRU. The performance (final BW, weight gain and gain per area) of the animals was different, being higher (p = 0.04) in animals supplemented with soybean meal, compared to NPN. The removal of soybean meal from the supplement and its replacement with either conventional urea plus post-ruminal urea or only post-ruminal urea compromises the performance of the animals. The lower the post-ruminal urea inclusion level, the lower the apparent digestibility of dry matter, organic matter, and NDF, when compared to animals supplemented with higher levels. [ABSTRACT FROM AUTHOR]

The effects of different inclusion rates of Poncirus trifoliata (PT) extract on in vitro ruminal fermentation parameters were investigated. Six dose levels were referred to as follows: PT0, control (without PT); PT1, (3 mg/L); PT3, (9 mg/L); PT5, (15 mg/L); PT7, (21 mg/L); PT9, (27 mg/L), each incubated with 50 mL of buffered rumen fluid and 300 mg of timothy hay as a substrate for 72 h of incubation using an in vitro batch culture system. The in vitro dry matter digestibility (IVDMD) was significantly affected by the PT dose, except after 6 and 48 h of incubation. Total volatile fatty acids (VFAs) increased quadratically (12 h of incubation) within the range of PT1–PT7, whereas decreased at PT9 dose. Methane (CH4) production and the proportion of CH4 in the total gas produced decreased linearly after 12 h of incubation. The relative abundance of Ruminococcus albus and Ruminoccocus flavefaciens was greater at the doses PT1 than PT0, whereas that of Fibrobacter succinogenes was greater at PT0 than at PT9 after 24 h of incubation. Methanogenic archaea decreased linearly with increasing PT dose after 12 h of incubation. The abundance of ciliate-associated methanogens decreased linearly over 24 h of incubation under PT7 and PT9 treatments. These results suggest that the dose of PT can modulate ruminal fermentation to alter VFA concentration and reduce CH4 production by altering ruminal bacterial abundance. This study evaluated the dose-response effects of Poncirus trifoliata as a methane mitigation agent in ruminants. Inclusion of Poncirus trifoliate lowered in vitro dry matter digestibility and total volatile fatty acid concentration. Inclusion of Poncirus trifoliata extract reduced methane production after 12 h of incubation but did not maintained. Inclusion of Poncirus trifoliata extract reduced methanogenic archaea and ciliate-associated methanogens abundance [ABSTRACT FROM AUTHOR]

Mulberry flavonoids can modulate the composition of rumen microbiota in ruminants to improve nutrient digestibility, owing to their strong biological activities. This study aimed to explore the effect of mulberry leaf flavonoids (MLF) on rumen bacteria, fermentation kinetics, and metagenomic functional profile in water buffalo. Forty buffaloes (4 ± 1 lactations) with almost same body weight (av. 600 ± 50 Kg) and days in milk (90 ± 20 d) were randomly allocated to four treatments having different levels of MLF: 0 g/d (control), 15 g/d (MLF15), 30 g/d (MLF30), and 45 g/d (MLF45) supplemented in a basal diet. After 35 days of supplementation, rumen contents were collected to determine rumen fermentation parameters. The 16S rRNA gene sequencing was performed to elucidate rumen bacteria composition. The obtained taxonomic data were analyzed to explore the rumen bacteriome and predict the associated gene functions and metabolic pathways. Results demonstrated a linear increase (p < 0.01) in rumen acetate, propionate, and total VFAs in the MLF45 group as compared to control. No effect of treatment was observed on rumen pH and butyrate contents. Acetate to propionate ratio in the MLF45 group linearly and quadratically decreased (p = 0.001) as compared to MLF15 and control groups. Similarly, MLF45 linearly increased (p < 0.05) the microbial protein (MCP) and NH3-N as compared to other treatments. Treatment adversely affected (p < 0.01) almost all alpha diversity parameters of rumen bacteria except Simpson index. MLF promoted the abundance of Proteobacteria while reducing the relative abundances of Actinobacteria, Acidobacteria, Chloroflexi, and Patescibacteria. The MLF supplementation tended to substantially reduce (0.05 < p < 0.1) the abundance of Actinobacteria, and Patescibacteria while completely eliminating Acidobacteria (p = 0.029), Chloroflexi (p = 0.059), and Gemmatimonadetes (p = 0.03) indicating the negative effect of flavonoids on the growth of these bacteria. However, MLF45 tended to substantially increase (p = 0.07) the abundance (~21.5%) of Acetobacter. The MLF treatment exhibited negative effect on five genera by significantly reducing (Sphingomonas) or eliminating (Arthobactor, unclassified_c__Actinobacteria, norank_c__Subgroup_6, norank_o__Saccharimonadales, and Nocardioides) them from the rumen microbiota. Pearson correlation analysis revealed 3, 5 and 23 positive correlations of rumen bacteria with milk yield, rumen fermentation and serum antioxidant parameters, respectively. A positive correlation of MCP was observed with three bacterial genera (Acetobacter, Enterobacter, and Klebsiella). The relative abundance of Pseudobutyrivibrio and Empedobacter also showed a positive correlation with the ruminal acetate and propionate. The present study indicated 45 g/d as an appropriate dose of MLF which modulated rumen bacteria and its functional profile in water buffalo. [ABSTRACT FROM AUTHOR]

Background and Aim: Understanding the regulations of rumen microbiota and their fibrolytic capabilities under different forages are essential to improve rumen fermentation and animal feed efficiency. This study aimed to evaluate the changes in the rumen fermentation and the structure and fibrolytic activities of rumen bacteria in camels fed barley straw and Egyptian clover hay. Materials and Methods: Three fistulated camels were fed a diet containing barley straw for 30 days; then transitioned to a diet containing Egyptian clover hay for 30 days. In addition, bacterial media enriched with xylan and different cellulose sources, namely, filter paper, wheat straw, and alfalfa hay, were used to evaluate the ability of camel rumen bacteria to produce xylanase and cellulase enzymes. Results: The camel group fed Egyptian clover hay showed higher crude protein intake, rumen ammonia, total volatile fatty acids, and acetic acid. Moreover, the camel group fed barley straw showed higher neutral detergent fiber intake, rumen pH, and propionic and butyric acids. Principal component analysis showed that bacterial communities were separated based on the forage type. Forage type affected the composition of rumen bacteria and most of the bacterial community was assigned to phylum Bacteroidetes and Firmicutes. Egyptian clover hay diet increased the proportions of genus Prevotella and Ruminococcus; while fed barley straw diet increased the Butyrivibrio, RC9_gut_group, and Fibrobacteres. The bacterial culture of the Egyptian clover hay fed group produced the greatest xylanase and the bacterial culture of the barley straw fed group produced the maximum cellulase. Conclusion: Egyptian clover hay is recommended to feed camels in intensive production. Moreover, the bacterial community in the camel rumen is a promising source of lignocellulolytic enzymes.

This study examined the effect of the ratio of dietary metabolizable energy (MJ) to nitrogen (g) content (ME:N) on average daily gain (ADG), blood biochemical indices, rumen fermentation parameters, and rumen bacterial community in yaks. Thirty-six male yaks, aged 2-3 years, were divided into three groups and received a ME:N ratio of 0.42 (HY), 0.36 (MY,) or 0.32 (LY) MJ/g. Dry matter intake ranged between 3.16 and 3.63 kg/d and was lesser (p < 0.001) in the LY group than the other two groups. ME intake increased (p < 0.001) with an increase in the ME:N ratio, while N intake did not differ among groups. The ADG was 660 g/day for the MY group, which was higher (p < 0.005) than the 430 g/day in the LY group, while the HY group gained 560 g/day and did not differ from the other two groups. Feed intake to ADG ratio ranged between 5.95 and 7.95, and numerically was highest in the LY group and lowest in the MY group. In general, the concentration of ruminal total volatile fatty acids (p < 0.03) and molar proportions of propionate (p < 0.04), increased, while the molar proportion of acetate (p < 0.005) and the acetate:propionate ratio decreased (p < 0.001) with a decrease in the ME:N ratio. The molar proportion of butyrate did not differ among groups (p = 0.112). Group MY had higher ruminal NH3-N content than group HY and had a higher serum glucose content but lower urea content, lactate dehydrogenase, and creatine kinase content than group LY. In ruminal bacteria at the phylum level, the relative abundance of Firmicutes (F) was greater and of Bacteroidetes (B) was lesser, while the F:B ratio was greater in group MY than in groups HY an LY. We concluded that the yaks consuming the diet containing a ME:N ratio of 0.36 MJ/g had the best performance of the three groups. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effect of Allium mongolicum Regel ethanol extract (AME) on the concentration of three branched-chain fatty acids (BCFAs) related to flavor, fermentation parameters and the bacteria and their correlations in the rumen of lambs. A total of thirty 3-month-old male, Small-tailed Han sheep (33.60 ± 1.23 kg) were randomly distributed into 2 groups as follows: control group (CON) was fed a basal diet and AME group was fed a basal diet supplemented with 2.8 g.lamb-1.d-1 A. mongolicum Regel ethanol extract. AME supplementation decreased (P = 0.022) 4-methyloctanoic acid (MOA) content and tended to lower (P = 0.055) 4-methylnonanoic acid (MNA) content in the rumen. Compared to CON group, the ruminal concentrations of valerate and isovalerate were higher (P = 0.046 and P = 0.024, respectively), and propionate was lower (P = 0.020) in the AME group. At the phylum level, the AME group had a lower abundance of Bacteroidetes (P = 0.014) and a higher abundance of Firmicutes (P = 0.020) than the CON group. At the genus level, the relative abundances of Prevotella (P = 0.001), Christensenellaceae_R-7_group (P = 0.003), Succiniclasticum (P = 0.004), and Selenomonas (P = 0.001) were significantly lower in the AME group than in the CON group, while the relative abundances of Ruminococcus (P < 0.001), Quinella (P = 0.013), and Lachnospiraceae_XPB1014_group (P = 0.001) were significantly higher. The relative abundances of Prevotella (P = 0.029, R = 0.685; P = 0.009, R = 0.770), Christensenellaceae_R-7_group (P = 0.019, R = 0.721; P = 0.029, R = 0.685), and Succiniclasticum (P = 0.002, R = 0.842; P = 0.001, R = 0.879) was positively correlated with MOA and MNA levels, and the relative abundance of Lachnospiraceae_XPB1014_group (P = 0.033, R = -0.673) was negatively correlated with MOA. The relative abundance of Christensenellaceae_R-7_group (P = 0.014, R = -0.744) and Prevotellaceae_UCG-003 (P = 0.023, R = -0.706) correlated negatively with the EOA content. In conclusion, these findings suggest that the AME affected the concentration of BCFAs, fermentation parameters and the rumen bacteria in the rumen of lambs. [ABSTRACT FROM AUTHOR]

Simple Summary: During the antler-growing period, sika deer (Cervus nippon) need a large amount of protein from feed for rapid antler growth. Antlers are the most important economic product of sika deer. Methionine is the first or second limiting amino acid in the diets of sika deer, which greatly limits the effective utilization of protein in the feed, while methionine has biological functions such as antioxidant and immune function improvement. In this study, we evaluated for the first time the effect of methionine supplementation in the diet on antler-growing sika deer and tried to investigate the changes in rumen microorganisms and obtain better production efficiency through methionine regulation. The results showed that supplementation with appropriate methionine improved the antioxidant and immune function of the sika deer, while obtaining better antler weight. Methionine is the first or second limiting amino acid for ruminants, such as sika deer, and has a variety of biological functions such as antioxidant activity, immune response, and protein synthesis. This study aimed to investigate the effects of methionine supplementation on antler growth, serum biochemistry, rumen fermentation, and the bacterial community of sika deer during the antler-growing period. Twelve 4-year-old male sika deer were randomly assigned to three dietary groups supplemented with 0 g/day (n = 4, CON), 4.0 g/day (n = 4, LMet), and 6.0 g/day (n = 4, HMet) methionine. No significant difference (p > 0.05) was found in the production performance between the three groups, but antler weight was higher in both the LMet and HMet groups than in the CON group. Methionine supplementation significantly increased the serum glutathione peroxidase activity (p < 0.05). The serum immunoglobulin G level was significantly higher in the HMet group than in the other two groups (p < 0.05). No significant effect was found on the apparent amino acid digestibility of the three groups, but cysteine and methionine digestibility were higher in the LMet group. The serum hydroxylysine level was significantly lower in the LMet and HMet groups, whereas the serum lysine level was significantly lower in the HMet group compared with the CON group (p < 0.05). The LMet group had the highest but a nonsignificant total volatile fatty acid content and significantly higher microbial protein content in the rumen than the CON group (p < 0.05). The phyla Bacteroidetes, Firmicutes, and Proteobacteria were dominant in the rumen of the sika deer. The principal coordinate analysis (PCoA) and analysis of similarities (ANOSIM) results showed a significant change in the bacterial composition of the three groups (p < 0.05). The relative abundance of Prevotella and Rikenellaceae-RC9 was significantly higher in the LMet group compared with the CON group and CON and HMet groups, respectively. These results revealed that methionine supplementation improved the antioxidant activity and immune status, affecting amino acid metabolism and rumen microbial composition of the sika deer. [ABSTRACT FROM AUTHOR]

Soybean peptides (SPs), a feed additive derived from soybean, exhibit nutritional function and biological activity in monogastric animals, but limited studies have been conducted in dairy cows. Our experiments were conducted to evaluate the effects of SPs on the nutrient degradability of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), and acid detergent fiber (ADF) in vitro and milk production, rumen fermentation and bacterial community, and blood parameters of dairy cows. For in vitro experiment, ruminal fluids were collected from three ruminal cannulated Holstein dairy cows. A total of three levels of SPs (0, 0.38, and 1.92 g/kg DM of SPs) were added to the total mixed ration (TMR). Nutrient degradability and fermentation fluid pH were determined at 24 and 48 h using 3.0 g samples of the substrate. Gas production after 48 h was recorded by an automated trace gas recording system using 0.5 g samples of the substrate. The results showed that DM, NDF, ADF (p < 0.01), and CP (p < 0.05) degradabilities were significantly increased at 1.92 g/kg DM of SPs at 24 h, and asymptotic gas production (p = 0.05) was increased at 48 h. For in vivo experiment, 110 lactating Holstein cows (209.7 ± 65.2 DIM; 37.2 ± 6.4 kg/d milk yield) were randomly assigned to 0 (control group, CON) or 50 g/head/day SPs (SP-supplemented group). Yields of milk (p < 0.05), milk protein (p < 0.05), and milk lactose (0.05 < p < 0.10) increased on SPs supplementation; however, the milk fat percentage decreased (p < 0.05). The concentrations of individual volatile fatty acids (VFAs) (p < 0.05) and superoxide dismutase (SOD) (p < 0.01) were also increased. Rumen bacterial diversity in SP-supplemented cows was higher (p < 0.05). The relative abundances of Rikenellaceae_RC9_gut_group, Butyrivibrio, Selenomonas, and Shuttleworthia were significantly increased and that of Coprococcus was decreased (p < 0.05). Our results showed that supplementing 1.92 g/kg DM of SPs could improve the nutrient degradability in vitro and 50 g/head/day of SPs could improve milk production and antioxidant ability of dairy cows. The rumen bacterial diversity was also enhanced by SP supplementation.

Precision dietary interventions (e.g., altering proportions of dietary protein fractions) has significant implications for the efficiency of nutrient use in ruminants, as well as lowering their environmental footprint, specifically nitrogen (N) emissions. Soluble protein (SP) is defined as the protein fraction that is rapidly degraded in the rumen (e.g., non-protein N and true protein), and our previous study found that regulating SP levels could improve N efficiency in Hu sheep. Thus, the present study was conducted to explore in vitro how protein fractions with different SP levels modulate the rumen microbial community and its association with N metabolism. Four dietary treatments with different SP proportions and similar crude protein (CP) content (~14%) were formulated (% of CP): 20 (S20), 30 (S30), 40 (S40) and 50 (S50). Results showed that NH3-N content increased with increasing SP levels at 4, 12 and 24 h; TVFA, acetate, propionate and valerate were higher in S30 and S40 (p < 0.05) and had quadratic effects (p < 0.05). Moreover, dry matter digestibility (DMD) and N digestibility (ND) were all decreased with S20 and S50 (p < 0.05). The S30 and S40 treatments increased the abundance of Bacteroidetes and Prevotella (Prevotella_ruminicola) but decreased the abundance of Firmicutes and Proteobacteria (p < 0.05). Bacterial pathways related to amino acid and fatty acid metabolism also were enriched with S30 and S40. The abundance of Entodinium was increased with S30 and S40 and had a positive correlation with Prevotella, and these two genera also played an important role in N metabolism and VFA synthesis of this study. In conclusion, bacterial and protozoal communities were altered by the level of SP (% of CP), with higher SP levels (~50% of CP) increasing the microbial diversity but being detrimental to rumen N metabolism. [ABSTRACT FROM AUTHOR]

Bacteria are the most abundant microorganisms in the rumen microbiota and play essential roles, mainly fermenting plant compounds that yield fatty acids. In this study, we aimed at assessing stability of both bacterial composition and of its associations with rumen and milk fatty acids phenotypes over a 1-week period. The study was performed using 118 Lacaune dairy ewes from the INRAE Experimental Unit of La Fage. Rumen and milk samples were obtained from the ewes twice, 1 week apart, and microbiota composition, volatile and long-chain fatty acid concentrations were analyzed. Bacterial composition was assessed using 16S rRNA gene sequencing, and microbiota and fatty acids were analyzed as compositional data. As we worked with relative abundances expressed in a constrained space, the centered log-ratio transformation enabled to transform data to work with multivariate analyses in the Euclidian space. Bacterial composition differed between the 2 weeks of sampling, characterized by different proportions of the two main phyla, Bacteroidetes and Firmicutes. The repeatability of the operational taxonomic units (OTUs) was low, although it varied significantly. However, 66 of them presented a repeatability of over 0.50 and were particularly associated with fatty acid phenotypes. Even though the OTUs from the same bacterial families presented similar correlations to fatty acids in both weeks, only a few OTUs were conserved over the 2 weeks. We proved with the help of sequencing data that there is significant change in microbial composition over a week in terms of abundance of different families of bacteria. Further studies are required to determine the impact of bacterial composition alterations over 1 week, and the specificities of the highly repeatable OTUs. [ABSTRACT FROM AUTHOR]

Objective The objective of this study was to compare fenugreek (FG) with alfalfa (Alf) in ruminal fermentation and methane (CH4) production in vitro. Methods Whole-plant FG harvested at 11- and 15-wk and Alf harvested at early and mid-bloom maturities, alone or as 50:50 mixture of FG and Alf at the respective maturity, were assessed in a series of 48-h in vitro batch culture incubations. Total fermentation gas and methane gas production, dry matter (DM) disappearance, volatile fatty acids, microbial protein and 16S RNA gene copy numbers of total bacteria and methanogens were determined. Results Compared to early bloom Alf, FG harvested at 11-wk exhibited higher (p

Bacteria are the most abundant microorganisms in the rumen microbiota and play essential roles, mainly fermenting plant compounds that yield fatty acids. In this study, we aimed at assessing stability of both bacterial composition and of its associations with rumen and milk fatty acids phenotypes over a 1-week period. The study was performed using 118 Lacaune dairy ewes from the INRAE Experimental Unit of La Fage. Rumen and milk samples were obtained from the ewes twice, 1 week apart, and microbiota composition, volatile and long-chain fatty acid concentrations were analyzed. Bacterial composition was assessed using 16S rRNA gene sequencing, and microbiota and fatty acids were analyzed as compositional data. As we worked with relative abundances expressed in a constrained space, the centered log-ratio transformation enabled to transform data to work with multivariate analyses in the Euclidian space. Bacterial composition differed between the 2 weeks of sampling, characterized by different proportions of the two main phyla, Bacteroidetes and Firmicutes. The repeatability of the operational taxonomic units (OTUs) was low, although it varied significantly. However, 66 of them presented a repeatability of over 0.50 and were particularly associated with fatty acid phenotypes. Even though the OTUs from the same bacterial families presented similar correlations to fatty acids in both weeks, only a few OTUs were conserved over the 2 weeks. We proved with the help of sequencing data that there is significant change in microbial composition over a week in terms of abundance of different families of bacteria. Further studies are required to determine the impact of bacterial composition alterations over 1 week, and the specificities of the highly repeatable OTUs.

Antimicrobial peptides (AMPs) are gaining popularity as alternatives for treatment of bacterial infections and recent advances in omics technologies provide new platforms for AMP discovery. We sought to determine the antibacterial activity of a novel antimicrobial peptide, buwchitin, against Enterococcus faecalis. Buwchitin was identified from a rumen bacterial metagenome library, cloned, expressed and purified. The antimicrobial activity of the recombinant peptide was assessed using a broth microdilution susceptibility assay to determine the peptide's killing kinetics against selected bacterial strains. The killing mechanism of buwchitin was investigated further by monitoring its ability to cause membrane depolarization (diSC3(5) method) and morphological changes in E. faecalis cells. Transmission electron micrographs of buwchitin treated E. faecalis cells showed intact outer membranes with blebbing, but no major damaging effects and cell morphology changes. Buwchitin had negligible cytotoxicity against defibrinated sheep erythrocytes. Although no significant membrane leakage and depolarization was observed, buwchitin at minimum inhibitory concentration (MIC) was bacteriostatic against E. faecalis cells and inhibited growth in vitro by 70% when compared to untreated cells. These findings suggest that buwchitin, a rumen derived peptide, has potential for antimicrobial activity against E. faecalis.