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The objective of this study was to systematically investigate how sodium butyrate (SB) affects the gastrointestinal bacteria in newborn calves at different stages before weaning. Forty female newborn Holstein calves (4-day-old, 40 ± 5 kg of body weight) were randomly divided into four groups; each group was supplemented with four SB doses: 0, 15, 30, and 45 g/day (ten replicates) in SB0, SB15, SB30, and SB45 groups, respectively. SB was fed with milk replacer from day 4 to day 60. Rumen fluid and feces were collected on days 2, 14, 28, 42, and 60 for 16S rRNA high-throughput sequencing. Data were analyzed in a complete randomized design and analyzed on the online platform of Majorbio Cloud Platform. The results showed that SB significantly increased the α-diversity in feces, especially Shannon and Chao indices in SB45 and SB30 at day 60 more than in SB15 (P < 0.05). Additionally, SB significantly enhanced Firmicutes growth from day 2 to 28 and also increased Bacteroides abundance from day 28 to 42 in rumen and feces (P < 0.05). SB also significantly inhibited Proteobacteria abundance in rumen and feces during the study period (P < 0.05). SB also promoted some potential beneficial bacterial abundance, including Prevotella, Lachnospiraceae, Clostridium, Ruminococcus, and Muribaculaceae (P < 0.05). Additionally, Escherichia-Shigella abundance at SB0 was significantly lower than in the other groups (P < 0.05). In conclusion, this study firstly reported a dynamic curve showing of the SB effects on bacteria in calves before weaning. This study provides valuable evidence for the development of the gastrointestinal tract of the calves in the early stage of the life. SB supplementation improved the gastrointestinal health by regulating the bacterial populations.Key points• The gastrointestinal tract of calves has been improved after the SB supplementation.• Microbes were the vital influential factor in the development of calves.• Intervention before weaning is an effective strategy for calf health.

Digestibility trials need a viable rumen fluid as inoculum to degrade feeds. The variability of rumen fluid depends on the animal’s diet, while its viability is greatly influenced by the sampling and handling procedures. In this article, we present a replicable protocol for sampling the rumen fluid from slaughtered animals for in vitro digestibility trials. A detailed list of the tools and a step-by-step standardized procedure for the collection, storage and the transportation of the rumen fluid from the slaughterhouse to the laboratory is presented. We also describe a digestibility trial for establishing the maximum storage time of rumen fluid from sampling to its use. The results show that the rumen fluid, collected and maintained according to the proposed protocol, can be stored and used from 30 to 300 min from sampling without significantly compromising the fermentative activity of the microbial population.

Abstract Background Rumen juice analysis (RJA) involves analysis of a 10mL sample within minutes after sampling. However, it can be challenging to collect 10 mL of rumen juice (RJ) from some ruminants, and clinical circumstances can delay RJA. Objectives Quantify the effect of sample volume (2, 5, 10, 50, and 100 mL), and time-to-analysis (0, 30, and 60 minutes) on RJA. Animals Cannulated cow. Methods Observational experimental study. Two liters of RJ were collected at 26 separate times. The samples were subdivided into 2 duplicates of each sample volume at each sampling time; and analyzed at 0, 30, and 60 minutes after collection. Rumen juice analysis included pH measurement, methylene blue reduction time (MBRT), and protozoal motility. Results The pH of 2 and 5 mL samples was significantly (P = .01) higher than the pH of 50 and 100 mL samples at all time points. The MBRT was significantly lower (faster bacterial reduction) for 100 mL samples compared to all other samples at 0 minute and to 2, 5, and 50 mL samples at 30 min. The pH and MBRT at 60 minutes were significantly higher than at 0 minute for all volumes (P < .05 and P < .01, respectively). For large protozoa, small sample volumes (2 and 5 mL) had significantly lower protozoal motility (scores of 5 and 4.5, respectively) compared to 100 mL samples at 60 minutes (score of 4; P < .05). Conclusions and Clinical Importance Interpretation of RJA could be affected by small sample volumes and delays to analysis. Sample volumes of ≥10 mL analyzed within 30 minutes after collection are recommended.

To identify differences in rumen function as a result of feeding monensin to beef cattle, rumen fluid metagenomics and metabolomics analyses were used to evaluate the functional attributes and metabolites of rumen microbiota in beef steers fed no or 200 mg/d of monensin. Eight rumen-fistulated steers were used in the study for a period of 53 days. Rumen fluid samples were collected on the last day of the experiment. Monensin increased the relative abundance of Selenomonas sp. ND2010, Prevotella dentalis, Hallella seregens, Parabacteroides distasonis, Propionispira raffinosivorans, and Prevotella brevis, but reduced the relative abundance of Robinsoniella sp. KNHs210, Butyrivibrio proteoclasticus, Clostridium botulinum, Clostridium symbiosum, Burkholderia sp. LMG29324, and Clostridium butyricum. Monensin increased the relative abundance of functional genes involved in amino acid metabolism and lipid metabolism. A total of 245 metabolites were identified. Thirty-one metabolites were found to be differentially expressed. Pathway analysis of the differentially expressed metabolites revealed upregulated metabolic pathways associated with metabolism of linoleic acid and some amino acids. These findings confirm that monensin affects rumen fermentation of forage-fed beef cattle by modulating the rumen microbiome, and by reducing amino acid degradation and biohydrogenation of linoleic acid in the rumen.

Fermentative kitchen waste to produce high-value chemicals (e.g., acetic acid) has been investigated actively in the past decades. Creating an alkaline condition is widely used to improve the hydrolysis of polysaccharide and inhibit the methanogenesis, but this method significantly increases the overall cost. Herein, the present study investigated the bioaugmentation with rumen fluid to improve acetic acid production from kitchen waste at neutral condition via strengthening hydrolytic and acid-forming bacteria. Results showed that the highest acetic acid yield reached 1.52 g/L at rumen fluid and granular sludge ratio of 1:1. The proportion of acetic acid in volatile fatty acids (VFAs) has increased by 10% compared to control. Microbial community analysis revealed that bioaugmentation with rumen fluid increased the relative abundance of Prevotella and Rikenellaceae_RC9_gut_group which has the ability to degrade polysaccharides and produce acetic acid. Moreover, the proliferation of butyric acid producers (Clostridium_sensu_stricto_1 and Clostridium_sensu_stricto_7) were inhibited significantly, which was in agreement with high acetic acid proportion in VFAs. The bioaugmentation strategy and process optimization provided an energy and cost-saving method for acetic acid production from kitchen waste.

Rumen cud transfaunation re-establishes rumen micro environment and improves fermentation in recipient animals affected with digestive disorders. Preserving rumen cud or fluid will increase its availability for the treatment of rumen fermentation disorders, without having to maintain donor animals. Rumen fluid collected from healthy goats, fed standard ration having roughage 70% and concentrate 30%, was lyophilized (prefreezing -80 °C, 48 h; lyophilization -45 °C, 32 h) using 5% glycerol as cryoprotectant. The 16 S metagenome analysis of the lyophilized rumen fluid (LRF) revealed an abundance of Prevotella (33.2%). Selenomonas ruminantium (1.87%) and Megasphaera elsdenii (0.23%) were also present. Twenty-four goats having history of high grain feeding and exhibiting clinical symptoms of rumen fermentation disorders were randomly distributed into either one of the two treatment groups viz . , T 1  = oral administration of LRF 31 g/animal/day and T 2  = oral administration of sodium bicarbonate (SB) 15 g/animal/day. Post intervention LRF and SB, improved animal body condition, feed intake, fecal consistency, elevated the ruminal pH at 48 h, reduced propionate and lactate at 48 h, reduced total volatile fatty acids (TVFA) and ammonia nitrogen at 24 h. Significant reduction in serum blood urea nitrogen (BUN) and urea levels were observed even from 24 h post intervention irrespective of the treatments. LRF significantly improved acetate and decreased propionate production compared to SB. LRF at 7.5% (v/v) can thus be used to counteract ruminal fermentation disorders in goats sequel to high grain ration.

Alpaca ( Vicugna pacos ) is a South American camelid that was introduced recently to Egypt and considered a zoo animal in different country zoos. There are no available data regarding the normal values of rumen fluid and hematobiochemical constituents of alpacas bred in Egypt. The aim of this study was to estimate normal values for rumen fluid and blood constituents of alpaca and determine the influence of season and sex on these parameters under Egyptian circumstances. The study was conducted on seventeen (8 female and 9 male); 2–6 years apparently healthy alpacas. Rumen fluid and blood samples were taken from each animal (from August 2022 to February 2023) and divided into summer and winter samples. Rumen fluid constituents were influenced by seasonal changes and showed a significant ( p  < 0.05) increase in the total protozoal count in summer, while in winter, rumen fluid pH and ammonia values were significantly ( p  < 0.05) increased. For hematology, the effect of season was evident in the red blood cells count and packed cell volume, which increased significantly ( p  < 0.05) in winter. Regarding serum biochemistry, winter showed significant ( p  < 0.05) elevation in glucose, urea and magnesium levels, while summer had significantly ( p  < 0.05) increased chloride levels. The effect of sex was minimal, and only glucose and creatinine values showed significant ( p  < 0.05) increases in males compared to females. The effect of season was evident in rumen fluid, hematology and serum biochemical parameters of alpaca, while sex has minimal effect on these parameters. Rumen fluid and hematobiochemical constituent values of apparently healthy alpacas bred under Egyptian conditions were consistent with those in other countries, while slight differences were observed in total protozoal count and serum minerals. The data obtained in this study can be used as preliminary data for the rumen fluid and hematobiochemical constituents of alpacas kept under Egyptian circumstances.

Monitoring rumination activity is considered a useful indicator for the early detection of diseases and metabolic disorders. Accelerometer-based sensor systems provide health alerts based on individual thresholds of rumination times in dairy cows. Detailed knowledge of the relationship between sensor-based rumination times and rumen physiology would help detect conspicuous animals and evaluate the treatment’s success. This study aimed to investigate the association between sensor-based health alerts and rumen fluid characteristics in Holstein-Friesian cows at different stages of lactation. Rumen fluid was collected via a stomach tube from 63 pairs of cows with and without health alerts (ALRT vs NALRT). Pairs were matched based on the day of lactation, the number of lactations, and health criteria. Rumen fluid was collected during and after health alerts. The parameters of color, odor, consistency, pH, redox potential, sedimentation flotation time, and the number of protozoa were examined. Results showed differences between both groups in odor, rumen pH, sedimentation flotation time, and protozoan count at the first rumen fluid collection. Within the groups, greater variations in rumen fluid parameters were found for ALRT cows compared to NALRT cows. The interaction between health alert and stage of lactation did not affect the rumen fluid parameters.

Amino-acid-fermenting bacteria are wasteful organisms within the rumens of beef cattle that remove dietary amino nitrogen by producing ammonia, which is then excreted renally. There are currently no on-label uses for the control of this microbial guild, but off-label use of broad-spectrum antimicrobials has shown efficacy, which contributes to antimicrobial resistance. Plant-derived antimicrobials supplemented into the diets of cattle may offer worthwhile alternatives. This study sought to investigate the role of cannabidiol (CBD) as a terpenophenolic antimicrobial. Ex vivo cell suspensions were harvested from the rumen fluid of Angus × Holstein steers in non-selective media with amino acid substrates. The suspensions were treated with five concentrations of CBD (860 μg mL−1–0.086 μg mL−1) and incubated (24 h), after which ammonia production and viable number of cells per substrate and treatment were measured. The data demonstrated a ~10–15 mM reduction in ammonia produced at the highest concentration of CBD and negligible changes in the viable number of amino-acid-fermenting bacteria. CBD does not appear to be a biologically or economically viable terpenophenolic candidate for the control of amino acid fermentation in beef cattle.

This study aimed to explore the dynamic variations in fermentation characteristics, bacterial diversity and community composition at two preservation temperatures as preservation time extended. Six rumen fluid samples collected from high-grain feeding sheep were stored at −20 °C or −80 °C for 0 day, 7 days, 14 days, 30 days, 60 days, 120 days, and 240 days. The results showed that the current preservation temperature did not alter the fermentation characteristics, bacterial diversity and community composition (p > 0.05). The concentrations of ammonia, microbial crude protein, acetate, propionate, butyrate, valerate, and total volatile fatty acids were higher when stored at 60 days (p < 0.05). Preservation time had no influence on bacterial richness and evenness (p > 0.05), whilst the relative abundances of Bacteroidota and Prevotella were numerically higher when stored at 30 days, and the opposite results were observed regarding Firmicutes. Both principal co-ordinates analysis (PCoA) and non-metric multidimensional scaling (NMDS) showed clusters among treatments in terms of preservation time and preservation temperature. Analysis of similarities (ANOSIM) also revealed similarities between treatments (p > 0.05). This study indicates that most fermentation characteristics in rumen fluid were altered after a 60-day preservation, whilst the preservation time for rumen bacterial community profile alteration was 30 days. It is recommended to finish the sample determination of rumen fluid within 30 days. This study may assist decision-making regarding the practicable time for rumen fluid determination, as well as viable preservation conditions for inoculum used for in vitro fermentation testing.