Seasonal effects on rumen microbiome and enteric methane (CH4) emissions are poorly documented. In this study, 6 Holstein and 6 Jersey steers were fed the same total mixed ration diet during winter, spring, and summer seasons under a 2×3 factorial arrangement for 30 days per season. The dry matter intake (DMI), rumen fermentation characteristics, enteric CH4 emissions and rumen microbiota were analyzed. Holstein had higher total DMI than Jersey steers regardless of season. However, Holstein steers had the lowest metabolic DMI during summer, while Jersey steers had the lowest total DMI during winter. Jersey steers had higher CH4 yields and intensities than Holstein steers regardless of season. The pH was decreased, while ammonia nitrogen concentration was increased in summer regardless of breed. Total volatile fatty acids concentration and propionate proportions were the highest in winter, while acetate and butyrate proportion were the highest in spring and in summer, respectively, regardless of breed. Moreover, Holstein steers produced a higher proportion of propionate, while Jersey steers produced a higher proportion of butyrate regardless of season. Metataxonomic analysis of rumen microbiota showed that operational taxonomic units and Chao 1 estimates were lower and highly unstable during summer, while winter had the lowest Shannon diversity. Beta diversity analysis suggested that the overall rumen microbiota was shifted according to seasonal changes in both breeds. In winter, the rumen microbiota was dominated by Carnobacterium jeotgali and Ruminococcus bromii, while in summer, Paludibacter propionicigenes was predominant. In Jersey steers, Capnocytophaga cynodegmi, Barnesiella viscericola and Flintibacter butyricus were predominant, whereas in Holstein steers, Succinivibrio dextrinosolvens and Gilliamella bombicola were predominant. Overall results suggest that seasonal changes alter rumen microbiota and fermentation characteristics of both breeds; however, CH4 emissions from steers were significantly influenced by breeds, not by seasons.

Previous studies have focused on the rumen microbiome and enteric methane (CH4) emissions in dairy cows, yet little is known about steers, especially steers of dairy breeds. In the present study, we comparatively examined the rumen microbiota, fermentation characteristics, and CH4 emissions from six non-cannulated Holstein (710.33 ± 43.02 kg) and six Jersey (559.67 ± 32.72 kg) steers. The steers were fed the same total mixed ration (TMR) for 30 days. After 25 days of adaptation to the diet, CH4 emissions were measured using GreenFeed for three consecutive days, and rumen fluid samples were collected on last day using stomach tubing before feeding (0 h) and 6 h after feeding. CH4 production (g/d/animal), CH4 yield (g/kg DMI), and CH4 intensity (g/kg BW0.75) were higher in the Jersey steers than in the Holstein steers. The lowest pH value was recorded at 6 h after feeding. The Jersey steers had lower rumen pH and a higher concentration of ammonia-nitrogen (NH3-N). The Jersey steers had a numerically higher molar proportion of acetate than the Holstein steers, but the opposite was true for that of propionate. Metataxonomic analysis of the rumen microbiota showed that the two breeds had similar species richness, Shannon, and inverse Simpson diversity indexes. Principal coordinates analysis showed that the overall rumen microbiota was different between the two breeds. Both breeds were dominated by Prevotella ruminicola, and its highest relative abundance was observed 6 h after feeding. The genera Ethanoligenens, Succinivibrio, and the species Ethanoligenens harbinense, Succinivibrio dextrinosolvens, Prevotella micans, Prevotella copri, Prevotella oris, Prevotella baroniae, and Treponema succinifaciens were more abundant in Holstein steers while the genera Capnocytophaga, Lachnoclostridium, Barnesiella, Oscillibacter, Galbibacter, and the species Capnocytophaga cynodegmi, Galbibacter mesophilus, Barnesiella intestinihominis, Prevotella shahii, and Oscillibacter ruminantium in the Jersey steers. The Jersey steers were dominated by Methanobrevibacter millerae while the Holstein steers by Methanobrevibacter olleyae. The overall results suggest that sampling hour has little influence on the rumen microbiota; however, breeds of steers can affect the assemblage of the rumen microbiota and different mitigation strategies may be needed to effectively manipulate the rumen microbiota and mitigate enteric CH4 emissions from these steers.

Butyrate is known to play a significant role in energy metabolism and regulating genomic activities that influence rumen nutrition utilization and function. Thus, this study investigated the effects of an isolated butyrate-producing bacteria, Clostridium saccharobutylicum, in rumen butyrate production, fermentation parameters and microbial population in Holstein-Friesian cow. An isolated butyrate-producing bacterium from the ruminal fluid of a Holstein-Friesian cow was identified and characterized as Clostridium saccharobutylicum RNAL841125 using 16S rRNA gene sequencing and phylogenetic analyses. The bacterium was evaluated on its effects as supplement on in vitro rumen fermentation and microbial population. Supplementation with 10 6 CFU/ml Clostridium saccharobutylicum increased (p < 0.05) microbial crude protein, butyrate and total volatile fatty acids concentration but had no significant effect on NH 3-N at 24 h incubation. Butyrate and total VFA concentrations were higher (p < 0.05) in supplementation with 10 6 CFU/ml Clostridium saccharobutylicum compared with control, with no differences observed for total gas production, NH 3-N and propionate concentration. However, as the inclusion rate (CFU/ml) of C. saccharobutylicum was increased, reduction of rumen fermentation values was observed. Furthermore, butyrate-producing bacteria and Fibrobacter succinogenes population in the rumen increased in response with supplementation of C. saccharobutylicum, while no differences in the population in total bacteria, protozoa and fungi were observed among treatments. Overall, our study suggests that supplementation with 10 6 CFU/ml C. saccharobutylicum has the potential to improve ruminal fermentation through increased concentrations of butyrate and total volatile fatty acid, and enhanced population of butyrate-producing bacteria and cellulolytic bacteria F. succinogenes.

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The effects of Ptecticus tenebrifer and Tenebrio molitor (PTM), as alternative fishmeal, on the growth performance, haematological values, biochemical and immune response and gut microbial diversity of Cyprinus carpio L. were investigated. Two hundred and forty hatchery‐reared juvenile C. carpio were divided among four experimental diet groups, formulated with the insect replacements: Control—30% fishmeal; Treatment 1—20% fishmeal + 10% PTM; Treatment 2—10% fishmeal + 20% PTM; Treatment 3—30% PTM. C. carpio fed 30% PTM had the highest final length and the largest increase in length. The red blood cell count was the highest in 10% PTM (0.92 M/µL) and lowest in the control group (0.23 M/µL). Fusobacteria was the dominant gut bacterial phylum, followed by Bacteroidetes, Firmicutes and Proteobacteria. Cetobacterium somerae was the dominant gut bacterial species, and its relative abundance was significant in the 10% and 30% PTM treatments. The relative abundance of Bacteroides massiliensis was the highest in the control group while there was decline in the treatments with PTM. These results suggest that P. tenebrifer and T. molitor as a fishmeal replacement lower the biochemical oxygen demand and chemical oxygen demand in the culture tanks and increase the length and change the gut microbiome of C. carpio.

The effects of rumen buffer agents on ruminal fermentation parameters and bacterial community composition were determined using in vitro and in vivo experiments in three rumen-cannulated, high-concentrate fed Holstein Friesian dairy cows. Experiment 1 in vitro treatments included bentonite, calcium carbonate, calcium oxide, sodium bicarbonate, sodium sesquicarbonate, and processed coral, and unbuffered samples served as the control. Experiment 2 in vitro treatments were based on the formulation of various combinations of the buffer agents used in Experiment 1. Combinations were selected for the in vivo study based on their buffering ability. Calcium oxide, sodium bicarbonate, and sodium sesquicarbonate stabilized the ruminal pH and improved in vitro rumen fermentation. The combined buffer agents had a significant effect on pH, buffering capacity, total gas, and total volatile fatty acids. Firmicutes and Bacteroidetes were the dominant phyla in both treatments and the control. Ruminococcus and Prevotella were found to be the dominant genera. Ruminococcus bromii was predominant in the treatment group. Prevotella jejuni was more abundant in the control group compared to the treatment group, in which its abundance was very low. Ruminococcus flavefaciens and Intestinimonas butyriciproducens gradually increased in abundance as cows received treatment. Overall, a high-concentrate diet administered to cows induced adverse changes in ruminal pH; however, buffer supplementation enhanced ruminal fermentation characteristics and altered bacterial community, which could contribute to preventing ruminal acidosis.

Influence of dietary organic trace minerals on enteric methane emissions and rumen microbiota of heat-stressed dairy steers

Kimchi cabbage (Brassica rapa L.) by-products treated or untreated with calcium oxide and alkaline hydrogen peroxide as substitutional ingredient of total mixed ration for Holstein steers Running Title (within 10 words)Kimchi cabbage by-products as total mixed ration ingredient