Prevotella Ruminicola Research Articles

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

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

Effects of Combined Organic Selenium and Zinc Supplementation on In Vitro Ruminal Enzyme Activities and Relative Populations of Several Bacterial Species

Selenium (Se) and zinc (Zn) are essential animal microminerals. Combining Se and Zn (Se-Zn) as a feed additive in its influence on rumen fermentation patterns is still very limited, so further investigation is needed. The present study explored the supplementation impact of combined Se-Zn from organic sources on rumen enzyme activity and relative abundance of several bacterial species through an in vitro method. Five treatments, each with six replicates were used in the study. The first group treated without Se and Zn supplementation (T0, control), the second group treated with 0.3 ppm Se + 60 ppm Zn (T1), the third group treated with 0.45 ppm Se + 60 ppm Zn (T2), the fourth group treated with 0.3 ppm Se + 90 ppm Zn (T3), and the fifth group treated with 0.45 ppm Se + 90 ppm Zn (T4). The parameters observed included rumen microbial enzyme activities (carboxyl methyl cellulase, amylase, protease) and the relative abundance of rumen microbes (Ruminococcus sp., Ruminococcus flavefaciens, Ruminococcus albus, Streptococcus sp., Prevotella ruminicola, and Eubacterium ruminantium). Results indicated that carboxyl methyl cellulase (CMC-ase) and amylase activities raised in T2, T3, and T4 in comparison to T1 and T0 treatments. Protease activity and protein enzyme content increased in T2 compared to all treatments. The relative abundance of Ruminococcus sp. and Ruminococcus albus was higher in T2 and T3 compared to T0 treatment. Furthermore, an elevated Ruminococcus flavefaciens was indicated in T2 compared to other treatments. The T2, T3, and T4 led to higher abundances of Eubacterium ruminantium, Prevotella ruminicola, and Ruminococcus albus compared to T0 and T1. It is concluded that organic Se and Zn enhanced the relative abundance of several bacterial species and the activity of enzymes in the rumen; optimal results are recommended when combining 0.45 ppm Se + 60 ppm Zn.

Metagenomic sequencing identified microbial species in the rumen and cecum microbiome responsible for niacin treatment and related to intramuscular fat content in finishing cattle.

Niacin is one of the essential vitamins for mammals. It plays important roles in maintaining rumen microecological homeostasis. Our previous study indicated that dietary niacin significantly elevated intramuscular fat content (IMF) in castrated finishing steers. Whether niacin affects fat deposition by regulating the microbial composition and functional capacities of gastrointestinal microbiome has been unknown yet. In this study, 16 castrated Xiangzhong Black cattle were randomly assigned into either control group fed with a basal concentrate diet (n = 8) or niacin group fed with a basal concentrate diet added 1000 mg/kg niacin (n = 8). Seven rumen samples and five cecum content samples were randomly collected from each of control and niacin groups for metagenomic sequencing analysis. A total of 2,981,786 non-redundant microbial genes were obtained from all tested samples. Based on this, the phylogenetic compositions of the rumen and cecum microbiome were characterized. We found that bacteria dominated the rumen and cecum microbiome. Prevotella ruminicola and Ruminococcus flavefaciens were the most abundant bacterial species in the rumen microbiome, while Clostridiales bacterium and Eubacterium rectale were predominant bacterial species in the cecum microbiome. Rumen microbiome had significantly higher abundances of GHs, GTs, and PLs, while cecum microbiome was enriched by CBMs and AAs. We found a significant effect of dietary niacin on rumen microbiome, but not on cecum microbiome. Dietary niacin up-regulated the abundances of bacterial species producing lactic acid and butyrate, fermenting lactic acid, and participating in lipid hydrolysis, and degradation and assimilation of nitrogen-containing compounds, but down-regulated the abundances of several pathogens and bacterial species involved in the metabolism of proteins and peptides, and methane emissions. From the correlation analysis, we suggested that niacin improved nutrient digestion and absorption, but reduced energy loss, and Valine, leucine and isoleucine degradation of rumen microbiome, which resulted in the increased host IMF. The results suggested that dietary manipulation, such as the supplementation of niacin, should be regarded as the effective and convenient way to improve IMF of castrated finishing steers by regulating rumen microbiome.

Metagenomics reveals the temporal dynamics of the rumen resistome and microbiome in goat kids

BackgroundThe gut microbiome of domestic animals carries antibiotic resistance genes (ARGs) which can be transmitted to the environment and humans, resulting in challenges of antibiotic resistance. Although it has been reported that the rumen microbiome of ruminants may be a reservoir of ARGs, the factors affecting the temporal dynamics of the rumen resistome are still unclear. Here, we collected rumen content samples of goats at 1, 7, 14, 28, 42, 56, 70, and 84 days of age, analyzed their microbiome and resistome profiles using metagenomics, and assessed the temporal dynamics of the rumen resistome in goats at the early stage of life under a conventional feeding system.ResultsIn our results, the rumen resistome of goat kids contained ARGs to 41 classes, and the richness of ARGs decreased with age. Four antibiotic compound types of ARGs, including drugs, biocides, metals, and multi-compounds, were found during milk feeding, while only drug types of ARGs were observed after supplementation with starter feed. The specific ARGs for each age and their temporal dynamics were characterized, and the network inference model revealed that the interactions among ARGs were related to age. A strong correlation between the profiles of rumen resistome and microbiome was found using Procrustes analysis. Ruminal Escherichia coli within Proteobacteria phylum was the main carrier of ARGs in goats consuming colostrum, while Prevotella ruminicola and Fibrobacter succinogenes associated with cellulose degradation were the carriers of ARGs after starter supplementation. Milk consumption was likely a source of rumen ARGs, and the changes in the rumen resistome with age were correlated with the microbiome modulation by starter supplementation.ConclusionsOur data revealed that the temporal dynamics of the rumen resistome are associated with the microbiome, and the reservoir of ARGs in the rumen during early life is likely related to age and diet. It may be a feasible strategy to reduce the rumen and its downstream dissemination of ARGs in ruminants through early-life dietary intervention.ALjrPAe7hUkeePWTGYd3KaVideo

258 Investigating the Ruminal Metagenome of Neonatal Holstein Dairy Calves fed an Essential oil-Based Feed Additive

Abstract Antibiotics remain the most effective tool for treating and preventing animal diseases in livestock production. However, the widespread use of antibiotics in food animals, as well as concerns about antibiotic resistance among bacterial pathogens, have raised awareness of the need to limit their use. As neonatal dairy calves are very susceptible to gastrointestinal infections that can result in high mortality and morbidity, the need to find alternatives to antibiotics has become a priority. Although essential oils (EO) have emerged as promising anti-microbial alternatives, their potential effects on the rumen microbiome are still poorly understood due to the complexity of their chemical components and limited understanding of their mechanisms of action. To gain further insight into the effects of EO on the bacterial communities of pre-weaned calves, we investigated the rumen metagenome of neonatal Holstein dairy calves that had been supplemented with 3.75g/feeding of a blend of EO (carvacrol, caryophyllene, p-cymene, cineole, terpinene, and thymol) as part of a previous study. Genomic DNA extracted from rumen contents collected from one of the EO-supplemented animals was used for high throughput sequencing using an Illumina Miseq (2x250) platform. Raw sequence reads were assembled into genomic contigs using custom-written Perl scripts. Coding sequences (CDS) were identified and annotated using RAST, with assignment of open reading frames (ORFs) to metabolic pathways using KEGG pathways. After assembly, a total of 3,052 contigs between 279 and 259,890 nt in length were obtained. From this set, 571 contigs affiliated to Prevotella ruminicola, representing 18.7% of total contigs built, were further analyzed. Data mining for specific functions within categories such as ‘protein metabolism’, ‘carbohydrates metabolism’, and ‘amino acids and derivatives’ was performed. This analysis resulted in the identification of all the enzymes involved in glycolysis. Two outcomes were predicted for pyruvate: conversion to lactate through lactate dehydrogenases (EC 1.1.1.27, EC 1.1.1.28), and the production of formate and acetyl-CoA through the activity of pyruvate formate-lyase (EC 2.3.1.54). Acetate (acetate kinase, EC 2.7.2.1) and propionate (Propionate kinase, EC 2.7.2.15) were other predicted end products. In addition to glucose fermentation pathways, enzymes potentially involved in the utilization of plant-based secondary metabolites were also identified, such as enzymes involved in Quinate-Chorismate metabolism: 3-dehydroquinate dehydratase (EC 4.2.1.10), Shikimate 5-dehydrogenase I alpha (EC 1.1.1.25), Shikimate kinase (EC 2.7.1.71), 5-Enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.19), and Chorismate synthase (EC 4.2.3.5). Identification of such metabolic pathways suggests that perhaps the supplemented EO additive could be used as a precursor for the synthesis of aromatic amino acids. Together, these findings indicate that EO supplementation may benefit animal health and performance by promoting the growth of bacterial species that can metabolize compounds in EO additives.

The oxidoreductase activity of Rnf balances redox cofactors during fermentation of glucose to propionate in Prevotella

Propionate is a microbial metabolite formed in the gastrointestinal tract, and it affects host physiology as a source of energy and signaling molecule. Despite the importance of propionate, the biochemical pathways responsible for its formation are not clear in all microbes. For the succinate pathway used during fermentation, a key enzyme appears to be missing—one that oxidizes ferredoxin and reduces NAD. Here we show that Rnf [ferredoxin—NAD+ oxidoreductase (Na+-transporting)] is this key enzyme in two abundant bacteria of the rumen (Prevotella brevis and Prevotella ruminicola). We found these bacteria form propionate, succinate, and acetate with the classic succinate pathway. Without ferredoxin:NAD+ oxidoreductase, redox cofactors would be unbalanced; it would produce almost equal excess amounts of reduced ferredoxin and oxidized NAD. By combining growth experiments, genomics, proteomics, and enzyme assays, we point to the possibility that these bacteria solve this problem by oxidizing ferredoxin and reducing NAD with Rnf [ferredoxin—NAD+ oxidoreductase (Na+-transporting)]. Genomic and phenotypic data suggest many bacteria may use Rnf similarly. This work shows the ferredoxin:NAD+ oxidoreductase activity of Rnf is important to propionate formation in Prevotella species and other bacteria from the environment, and it provides fundamental knowledge for manipulating fermentative propionate production.

Effects of Italian ryegrass silage-based total mixed ration on rumen fermentation, growth performance, blood metabolites, and bacterial communities of growing Hanwoo heifers.

This study utilized Italian ryegrass silage (IRGS) - based total mixed ration (TMR) as feedstuff and evaluated its effects on rumen fermentation, growth performance, blood parameters, and bacterial community in growing Hanwoo heifers. Twenty-seven Hanwoo heifers (body weight [BW], 225.11 ± 10.57 kg) were randomly allocated to three experimental diets. Heifers were fed 1 of 3 treatments as follows: TMR with oat, timothy, and alfalfa hay (CON), TMR with 19% of IRGS (L-IRGS), and TMR with 36% of IRGS (H-IRGS). Feeding high levels of IRGS (H-IRGS) and CON TMR to heifers resulted in a greater molar proportion of propionate in the rumen. The impact of different TMR diets on the BW, average daily gain, dry matter intake, and feed conversion ratio of Hanwoo heifers during the growing period did not differ (p > 0.05). Furthermore, the blood metabolites, total protein, albumin, aspartate aminotransferase, glucose, and total cholesterol of the heifers were not affected by the different TMR diets (p > 0.05). In terms of rumen bacterial community composition, 264 operational taxonomic units (OTUs) were observed across the three TMR diets with 240, 239, and 220 OTUs in CON, L-IRGS, and H-IRGS, respectively. IRGS-based diets increased the relative abundances of genera belonging to phylum Bacteroidetes but decreased the abundances of genus belonging to phylum Firmicutes compared with the control. Data showed that Bacteroidetes was the most dominant phylum, while Prevotella ruminicola was the dominant species across the three TMR groups. The relative abundance of Ruminococcus bromii in the rumen increased in heifers fed with high inclusion of IRGS in the TMR (H-IRGS TMR). The relative abundance of R. bromii in the rumen significantly increased when heifers were fed H-IRGS TMR while P. ruminicola increased in both L-IRGS and H-IRGS TMR groups. Results from the current study demonstrate that the inclusion of IRGS in the TMR is comparable with the TMR containing high-quality forage (CON). Thus, a high level of IRGS can be used as a replacement forage ingredient in TMR feeding and had a beneficial effect of possibly modulating the rumen bacterial community toward mainly propionate-producing microorganisms.

Expanding the rumen Prevotella collection: The description of Prevotella communis, sp. nov. of ovine origin

27 strains representing eight new Prevotella species were isolated from rumen of a single sheep in eight weeks interval. One of the putative species encompassing the highest number of isolated strains which also exhibited some genetic variability in preliminary data, was then selected for description of a novel species. We examined six strains in genomic and phenotypic detail, two of which may actually be the same strain isolated nearly three weeks apart. Other strains formed clearly diverged intraspecies lineages as evidenced by core genome phylogeny and phenotypic differences. Strains of the proposed new Prevotella species are strictly saccharolytic as is usual for rumen Prevotella, and use plant cell-wall xylans and pectins for growth. However, the range of cell-wall polysaccharides utilised for growth is rather limited compared to rumen generalists such as Prevotella bryantii or Prevotella ruminicola and this extends also to the inability to utilise starch, which is unexpected for the members of the genus Prevotella. Based on the data obtained, we propose Prevotella communis sp. nov. to accommodate strain E1-9T as well as other strains with the similar properties. The proposed species is widespread: two other strains were previously isolated from sheep in Japan and is also common in metagenomic data of cattle and sheep rumen samples from Scotland and New Zealand. It was also found in a collection of metagenome-assembled genomes originating from cattle in Scotland. Thus, it is a ubiquitous bacterium of domesticated ruminants specialising in degradation of a somewhat restricted set of plant cell wall components.

Effects of Different Yeast Selenium Levels on Rumen Fermentation Parameters, Digestive Enzyme Activity and Gastrointestinal Microflora of Sika Deer during Antler Growth.

The aim of this experiment was to study the effects of different selenium supplemental levels on rumen fermentation microflora of sika deer at the velvet antler growth stage. A total of 20 5-year-old, healthy sika deer at the velvet antler growth stage with an average body weight of (98.08 ± 4.93) kg were randomly divided into 4 groups, and each group was fed in a single house. The SY1 group was the control group, and the SY2 group, SY3 group and SY4 group were fed a basal diet supplemented with 0.3, 1.2 and 4.8 mg/kg selenium, respectively. The pretest lasted for 7 days, and the formal trial period lasted for 110 days. The results show that: At the velvet antler growth stage, the digestibility of neutral detergent fiber and acid detergent fiber of sika deer in the SY2 group was significantly higher than that in the control group (p < 0.01). The digestibility of cellulose and crude fiber of sika deer in the SY2 group was significantly higher than those in the SY3 and SY4 groups (p < 0.01) and significantly higher than that in the control group (p < 0.05). The contents of acetic acid and propionic acid in the rumen fluid of sika deer in the SY2 group were significantly higher than those in the SY1 group (p < 0.05). Digestive enzyme analysis of rumen fluid at the velvet antler growth stage showed that the activity of protease in rumen fluid in the SY2 group was significantly lower than those in the SY1 group and SY4 group (p < 0.05). The relative abundance of Fibrobacter succinogenes in the SY2 group was significantly higher than that in the SY1 group (p < 0.05) and extremely significantly higher than those in the SY3 and SY4 groups (p < 0.01). Correlation analysis between yeast selenium level and bacterial abundance showed that the yeast selenium content in rumen fluid was significantly positively correlated with Butyrivibrio and Succiniclasticum (p < 0.01). Further verification of bacterial flora functioning showed that the SY2 group was more inclined to the degradation and utilization of fiber. In conclusion, 0.3 mg/kg selenium supplementation can increase the abundance of Prevotella ruminicola and Fibrobacter succinogenes in the rumen of sika deer and improve the degradation of fibrous substances by mediating the catabolite repression process.

Application of different proportions of sweet sorghum silage as a substitute for corn silage in dairy cows.

This experiment explored the effects of different proportions of sweet sorghum silage as a substitute for corn silage on dry matter intake (DMI), milk yield, milk quality, apparent digestibility, rumen fermentation parameters, serum amino acid profile, and rumen microbial composition of dairy cows. A total of 32 mid-lactation Holstein dairy cows with similar body weights and parities were randomly divided into four treatments: 100% corn silage +0% sorghum silage (CON), 75% corn silage +25% sorghum silage (CS1), 50% corn silage +50% sorghum silage (CS2), and 25% corn silage +75% sorghum silage (CS3). The milk yield was increased (linear, p = .048) as the proportion of sweet sorghum increased. Linear (p = .003) and quadratic (p = .046) increased effects were observed in milk fat as corn silage was replaced with sorghum silage. Compared with the CON diet group, the CS2 and CS3 diet groups had lower dry matter (DM) (linear, p < .001), ether extract (EE) (linear, p < .001), and gross energy (GE) (linear, p = .001) digestibility of the dairy cows. The ruminal fluid aspartate (Asp) level decreased (linear, p = .003) as the proportion of sweet sorghum increased. Linear (p < .05) and quadratic (p < .05) increased effects were observed for the contents of threonine (Thr), glycine (Gly), valine (Val), leucine (Leu), tyrosine (Tyr), and histidine (His) in rumen fluid with the replacement of corn silage with sorghum silage. Cows fed the CS3 diet had greater Faecalibacterium, Bacteroides, and Prevotella ruminicola content/copy number than those fed the CON diet (p < .05). In conclusion, feeding sorghum silage as a replacement for corn silage could increase the milk yield and fat, promote the growth of rumen microbes, and provide more rumen fluid amino acids for the body and microbial utilization. We believe that sorghum silage is feasible for dairy cows, and it is reasonable to replace corn silage with 75% sorghum silage.

Effects of Curcumin on Growth Performance, Ruminal Fermentation, Rumen Microbial Protein Synthesis, and Serum Antioxidant Capacity in Housed Growing Lambs.

This experiment was conducted to investigate growth performance, ruminal fermentation, rumen microbial protein synthesis, and serum antioxidant capacity with different doses of curcumin (CUR) included in the diet of housed growing lambs. Forty-eight four-month-old Dorper × Thin-tailed Han F1 crossbred male lambs (body weight = 20.89 ± 1.15 kg, age = 120 ± 10 days; mean ± SD) were randomly divided into four groups for a single-factor, completely randomized experiment. Treatments comprised the following: the basal diet supplemented with 0 (Control), 300 mg/kg (300 CUR), 600 mg/kg (600 CUR), or 900 mg/kg (900 CUR) CUR, respectively. The results showed that dietary CUR increased average daily gain (ADG), and the 300 CUR group evidenced the highest value. There were no significant effects on dry matter intake (DMI) and DMI/ADG. Lambs in the 300 CUR group showed higher totals of volatile fatty acids (VFA) and acetate than other groups, while decreased valerate was observed with supplementary CUR. The ruminal pH and ammonia N (NH3-N) concentration decreased with increasing CUR, with the greatest effect in the 300 CUR group. The quadratic effects were found in pectinase, carboxymethyl cellulose, and protease, with the greatest value in the 300 CUR group. The microbial populations of total bacteria and Ruminococcus albus also responded quadratically, and the methanogens, protozoan, and Fibrobacter succinogenes populations decreased linearly with increasing CUR. Lambs receiving additional CUR showed increased Prevotella ruminicola population. Microbial protein (MCP) synthesis was promoted by supplementary CUR. As supplementation with CUR increased, the serum activity of total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) was enhanced, with the greatest value in the 300 CUR group. In conclusion, dietary CUR improved ruminal fermentation, promoted rumen microbial protein (MCP) synthesis, and enhanced serum antioxidant activity, as well as promoting growth performance in housed growing lambs.

Effect of galangal essential oils on rumen microbial population and biodiversity on in vitro rumen fermentation

The study aimed to evaluate the effect of administering galangal essential oil (EO) on the abundance of rumen bacteria using the 16s rRNA method. The treatments included a control (no EO addition), galangal EO (30, 60, 120 µL), and cineole (5 µL). The treatments were assessed using a 48‐hour in vitro batch culture of rumen fluid containing a 60:40 ratio of forage to concentrate. For amplification of the prokaryotes (bacteria and archaea) in region V4, 16s rRNA primer 5’GTGCCAGCMGCCGCGTAA, GGACTACHVGGGTWTCTAAT3’ was employed. The data for rumen microbial abundance were analysed descriptively, while the data for rumen microbial diversity were obtained from the report on the Next Generation Sequencing Method. The microbial composition of each sample was tested for operational taxonomic units (OTUs) with a 97% identity rate on a valid label. The 16S rRNA gene sequencing yielded a total of 3,977 OTUs. Adding galangal and cineole EOs resulted in the same variation of the Shannon index. The population index (chao1 index) was highest when 60 µL of galangal EO was added, compared to 30 and 120 µL of galangal EO and cineole. In addition, providing 60 µL of galangal EO decreased the abundance of Prevotella ruminicola compared to the control and cineole doses. The addition of galangal EO also led to a decline in the number of Methanobacteriales. The population of the fibre‐degrading bacteria group (Ruminococcus albus and Ruminococcus flavefaciens) was higher in a dose of galangal EO than the control and cineole. Therefore, it can be concluded that the effective dose of galangal EO, i.e. 60 µL/300 mg (DM feed) in vitro, can reduce the abundance of Prevotella bacteria and methanogens.

Effect of Dietary Inclusion of Riboflavin on Growth, Nutrient Digestibility and Ruminal Fermentation in Hu Lambs

The study evaluated the influences of riboflavin (RF) supply on the growth performance, nutrient digestibility and ruminal fermentation in lambs. Forty-eight Hu lambs were randomly assigned into four groups receiving RF of 0, 15, 30 and 45 mg/kg dry mater (DM), respectively. Increasing RF supply did not affect the DM intake, but quadratically increased the average daily gain and linearly decreased feed conversion ratio. Total-tract DM, neutral detergent fibre, acid detergent fibre and crude protein digestibility increased quadratically. Rumen pH and propionate molar percentage decreased linearly, total volatile fatty acids concentration, acetate proportion and the ratio of acetate to propionate increased linearly, but ammonia nitrogen concentration was unchanged with increasing RF supply. Linear increases were observed on the activities of carboxymethyl-cellulase, xylanase, pectinase and protease, and the populations of bacteria, fungi, protozoa, dominant cellulolytic bacteria, Ruminobacter amylophilus and Prevotella ruminicola. Methanogens population was not affected by RF supplementation. The microbial protein amount and urinary total purine derivatives excretion increased quadratically. The results indicated that 30 mg/kg DM RF supply improved growth performance, rumen fermentation and nutrient digestion in lambs.

Interactions between Entodinium caudatum and an amino acid-fermenting bacterial consortium: fermentation characteristics and protozoal population in vitro.

Ruminal protozoa, especially entodiniomorphs, engulf other members of the rumen microbiome in large numbers; and they release oligopeptides and amino acids, which can be fermented to ammonia and volatile fatty acids (VFAs) by amino acid-fermenting bacteria (AAFB). Studies using defaunated (protozoa-free) sheep have demonstrated that ruminal protozoa considerably increase intraruminal nitrogen recycling but decrease nitrogen utilization efficiency in ruminants. However, direct interactions between ruminal protozoa and AAFB have not been demonstrated because of their inability to establish axenic cultures of any ruminal protozoan. Thus, this study was performed to evaluate the interaction between Entodinium caudatum, which is the most predominant rumen ciliate species, and an AAFB consortium in terms of feed degradation and ammonia production along with the microbial population shift of select bacterial species (Prevotella ruminicola, Clostridium aminophilum, and Peptostreptococcus anaerobius). From an Ent. caudatum culture that had been maintained by daily feeding and transfers every 3 or 4 days, the bacteria and methanogens loosely associated with Ent. caudatum cells were removed by filtration and washing. An AAFB consortium was established by repeated transfers and enrichment with casamino acids as the sole substrate. The cultures of Ent. caudatum alone (Ec) and AAFB alone (AAFB) and the co-culture of Ent. caudatum and AAFB (Ec + AAFB) were set up in three replicates and incubated at 39°C for 72 h. The digestibility of dry matter (DM) and fiber (NDF), VFA profiles, ammonia concentrations, pH, and microscopic counts of Ent. caudatum were compared among the three cultures. The co-culture of AAFB and Ent. caudatum enhanced DM degradation, VFA production, and Ent. caudatum cell counts; conversely, it decreased acetate: propionate ratio although the total bacterial abundance was similar between Ec and the Ec + AAFB co-culture after 24 h incubation. The ammonia production and relative abundance of C. aminophilum and P. anaerobius did not differ between AAFB alone and the Ec + AAFB co-culture. Our results indicate that Ent. caudatum and AAFB could have a mutualistic interaction that benefited each other, but their interactions were complex and might not increase ammoniagenesis. Further research should examine how such interactions affect the population dynamics of AAFB.

Altered gut microbiome composition by appendectomy contributes to colorectal cancer

Appendectomy impacts the homeostasis of gut microbiome in patients. We aimed to study the role of appendectomy in colorectal cancer (CRC) risk through causing gut microbial dysbiosis. Population-based longitudinal study (cohort 1, n = 129,155) showed a 73.0% increase in CRC risk among appendectomy cases throughout 20 years follow-up (Adjusted sub-distribution hazard ratio (SHR) 1.73, 95% CI 1.49–2.01, P < 0.001). Shotgun metagenomic sequencing was performed on fecal samples from cohort 2 (n = 314). Gut microbial dysbiosis in appendectomy subjects was observed with significant enrichment of 7 CRC-promoting bacteria (Bacteroides vulgatus, Bacteroides fragilis, Veillonella dispar, Prevotella ruminicola, Prevotella fucsa, Prevotella dentalis, Prevotella denticola) and depletion of 5 beneficial commensals (Blautia sp YL58, Enterococcus hirae, Lachnospiraceae bacterium Choco86, Collinsella aerofaciens, Blautia sp SC05B48). Microbial network analysis showed increased correlation strengths among enriched bacteria and their enriched oncogenic pathways in appendectomy subjects compared to controls. Of which, B. fragilis was the centrality in the network of the enriched bacteria. We further confirmed that appendectomy promoted colorectal tumorigenesis in mice by causing gut microbial dysbiosis and impaired intestinal barrier function. Collectively, this study revealed appendectomy-induced microbial dysbiosis characterized by enriched CRC-promoting bacteria and depleted beneficial commensals, signifying that the gut microbiome may play a crucial role in CRC development induced by appendectomy.

Identification of Mutations Responsible for Improved Xylose Utilization in an Adapted Xylose Isomerase Expressing Saccharomyces cerevisiae Strain

Economic conversion of biomass to biofuels and chemicals requires efficient and complete utilization of xylose. Saccharomyces cerevisiae strains engineered for xylose utilization are still considerably limited in their overall ability to metabolize xylose. In this study, we identified causative mutations resulting in improved xylose fermentation of an adapted S. cerevisiae strain expressing codon-optimized xylose isomerase and xylulokinase genes from the rumen bacterium Prevotella ruminicola. Genome sequencing identified single-nucleotide polymorphisms in seven open reading frames. Tetrad analysis showed that mutations in both PBS2 and PHO13 genes were required for increased xylose utilization. Single deletion of either PBS2 or PHO13 did not improve xylose utilization in strains expressing the xylose isomerase pathway. Saccharomyces can also be engineered for xylose metabolism using the xylose reductase/xylitol dehydrogenase genes from Scheffersomyces stipitis. In strains expressing the xylose reductase pathway, single deletion of PHO13 did show a significant increase xylose utilization, and further improvement in growth and fermentation was seen when PBS2 was also deleted. These findings will extend the understanding of metabolic limitations for xylose utilization in S. cerevisiae as well as understanding of how they differ among strains engineered with two different xylose utilization pathways.

Effects of excessive urea on rumen morphology and microbiota in Jianzhou Da'er goat (Capra hircus)

Urea is frequently used as a protein supplement in ruminant diets, but if used improperly, it can easily result in urea poisoning. The purpose of this study was to explore the effects of excessive urea feeding on rumen pathology and microbial diversity of the Jianzhou Da'er goat (Capra hircus). In this study, 9 male Jianzhou Da'er goats with an average weight of 22.9 kg were randomly divided into the control group (0% urea), 5% urea group and 10% urea group (dry matter basis, 3 goats per group). Morphological differences of rumen tissues were analyzed by Hematoxylin-Eosin staining and Prussian blue staining, and rumen microorganisms differences were analyzed by 16S rDNA sequencing on the Illumina Novaseq platform. Histopathological analysis showed that the length of rumen papilla in the 5% urea and 10% urea groups were shortened compared to the control group, and the thickness of the stratum corneum and muscular layer were significantly thinned (P < 0.05). 16S rRNA sequencing analysis indicated that microbial richness and diversity were significantly lower in goats fed on 5% urea or 10% urea, the abundance of Prevotella ruminicola was significantly decreased (P < 0.05), and the abundance of Ruminobacter amylophilus was significantly increased (P < 0.05), as compared with the control group. This study indicated that feeding 5% and 10% urea could damage rumen tissue morphology. Feeding 10% urea significantly reduced rumen microbial diversity and the abundance of Prevotella ruminicola, but increased the abundance of Ruminobacter amylophilus.

Effects of guanidinoacetic acid supplementation on lactation performance, nutrient digestion and rumen fermentation in Holstein dairy cows.

Considering the high energy demand of lactation and the potential of guanidinoacetic acid (GAA) addition on the increase in creatine supply for cows, the present study investigated the effects of 0, 0.3, 0.6 and 0.9g kg-1 dry matter (DM) of GAA supplementation on lactation performance, nutrient digestion and ruminal fermentation in dairy cows. The study used 40 mid-lactation multiparous Holstein cows and the study duration was 100 days. DM intake was not affected, but milk and milk component yields and feed efficiency increased linearly with increasing GAA addition. The total-tract digestibility of DM, organic matter, neutral detergent fibre, acid detergent fibre and non-fibre carbohydrates increased linearly and that of crude protein increased quadratically with increasing GAA addition. When the addition level of GAA increased, ruminal pH, molar percentages of butyrate, isobutyrate and isovalerate and the acetate-to-propionate ratio decreased linearly, and the total volatile fatty acids concentration and propionate molar percentage also increased linearly, whereas the acetate molar percentage and ammonia-N concentration were unaltered. The activities of fibrolytic enzymes, α-amylase and protease increased linearly. The populations of total bacteria, fungi, Ruminococcus albus, Fibrobacter succinogenes, Ruminococcus flavefaciens, Ruminobacter amylophilus and Prevotella ruminicola increased linearly, whereas protozoa and methanogens decreased linearly with increasing GAA addition. As for the blood metabolites, concentrations of glucose, urea nitrogen and methionine were unchanged, total protein, albumin, creatine and homocysteine increased linearly, and folate decreased linearly with increasing GAA supply. The results of the present study indicate that supplementation of GAA improved milk performance and rumen fermentation in lactating dairy cows. © 2022 Society of Chemical Industry.

The gut microbiome, mild cognitive impairment, and probiotics: A randomized clinical trial in middle-aged and older adults

Advancing age coincides with changes in the gut microbiome and a decline in cognitive ability. Psychobiotics are microbiota-targeted interventions that can result in mental health benefits and protect the aging brain. This study investigated the gut microbiome composition and predicted microbial functional pathways of middle-aged and older adults that met criteria for mild cognitive impairment (MCI), compared to neurologically healthy individuals, and investigated the impact of probiotic Lactobacillus rhamnosus GG (LGG) in a double-blind, placebo-controlled, randomized clinical trial. A total of 169 community-dwelling middle-aged (52-59 years) and older adults (60-75 years) received a three-month intervention and were randomized to probiotic and placebo groups. Participants were further subdivided based on cognitive status into groups with intact or impaired cognition and samples were collected at baseline and post supplementation. Microbiome analysis identified Prevotella ruminicola, Bacteroides thetaiotaomicron, and Bacteroides xylanisolvens as taxa correlated with MCI. Differential abundance analysis at baseline identified Prevotella as significantly more prevalent in MCI subjects compared to cognitively intact subjects (ALDEx2 P =0.0017, ANCOM-BC P =0.0004). A decrease in the relative abundance of the genus Prevotella and Dehalobacterium in response to LGG supplementation in the MCI group was correlated with an improved cognitive score. Our study points to specific members of the gut microbiota correlated with cognitive performance in middle-aged and older adults. Should findings be replicated, these taxa could be used as key early indicators of MCI and manipulated by probiotics, prebiotics, and symbiotics to promote successful cognitive aging. Registered under ClinicalTrials.gov Identifier no. NCT03080818.

Distinguishing the milk microbiota of healthy goats and goats diagnosed with subclinical mastitis, clinical mastitis, and gangrenous mastitis.

Mastitis, mainly caused by bacterial intramammary infections, is the main problem in the breeding of dairy animals. The inflammations of the mammary gland is separated by types of mastitis, being subclinical, clinical, and the most severe, gangrenous mastitis. Here, we used 16S rRNA amplicon sequencing to characterize the bacterial microbiota of goat milk in the different types of goat mastitis caused by bacteria. We used 72 goat milk samples from a region of the state of Minas Gerais in Brazil, of which 12 were from clinically healthy animals, 42 from animals diagnosed with subclinical mastitis, 16 from animals with clinical mastitis, and 2 from animals with gangrenous mastitis. The group related to gangrenous mastitis was the most divergent in terms of alpha and beta diversity. The most abundant genus among samples of the groups was Staphylococcus spp., and we found a high abundance of Mycoplasma sp. in the milk of animals diagnosed with clinical mastitis. The most statistically relevant microorganisms among the groups were Prevotella sp., Ruminococcaceae, Prevotella ruminicola sp., and Providencia sp. We highlight a new association of bacterial agents in gangrenous mastitis among Escherichia sp./Shigella sp. and Enterococcus sp. and provide the second report of the genus Alkalibacterium sp., in milk samples. Only the taxa Staphylococcus sp., Bacteroides sp., Enterococcus, and Brevidabacterium sp., were present in all groups. The superpathway of L-tryptophan biosynthesis metabolites and the sucrose degradation III (sucrose invertase) pathway were the most prominent ones among the groups. In this study, we demonstrate how a rich microbiota of goat milk from healthy animals can be altered during the aggravation of different types of mastitis, in addition to demonstrating new bacterial genera in milk not previously detected in other studies as well as new associations between agents.