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This study was aimed to evaluate the potential of a herbal mixture (HM) to improve production performance, rumen fermentation, and milk fatty acid profile in water buffaloes. Sixteen Murrah buffaloes (in four groups) were fed for 10 weeks with the same basal diet supplemented with 0 (control); 20 (HM20), 30 (HM30), and 40 (HM40) g/buffalo per day. The herbal mixture contained an equal quantity of black pepper (fruit), ginger (tubers), cinnamon (bark), peppermint (leaves), ajwain (seeds) and garlic (bulbs). After two weeks of adaptation, daily milk yield, and weekly milk composition were recorded. On the last day of the experiment, rumen contents were collected to determine rumen fermentation parameters and bacterial diversity through 16S rRNA sequencing. Results revealed no effect of treatment on dry matter intake (DMI), rumen fermentation parameters, and daily milk yield. However, milk fat (%) showed a tendency to increase (p = 0.07) in HM20 as compared with the control group. A significant increase in mono and polyunsaturated fatty acids (C14:1, C16:1, C18:2n6 and C18:3) whereas a decrease in saturated fatty acids (C18:0) in milk was observed in HM20 as compared with the control group. No significant change in bacterial diversity parameters (alpha and beta diversity) was observed in response to the treatment. Despite the substantial variation observed in the relative abundance of bacteria among treatment groups, no significant effect of treatment was observed when compared with the control group. Correlation analysis revealed several positive and negative correlations of rumen bacteria with rumen volatile fatty acids (VFA) and milk yield traits. Bacterial genera including Succinivibrionaceae, Butyrivibrio, Pseudobutyrivibrio, and Lachnospiraceae showed a positive correlation with VFA and milk yield traits. Overall, we observed 52 positive and 10 negative correlations of rumen bacteria with milk fatty acid contents. Our study revealed the potential of the herbal mixture at a lower supplemental level (20 g/day) to increase milk fat (%) and unsaturated fatty acid content in buffalo.

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]

Purpose: To investigate the effect of dietary supplementation of two omega fatty acids on in vitro rumen fermentation, microbial populations, total gas and methane (CH4) production． Methods: Both linoleic and linolenic acids were supplemented at 0 (control), 1, 3, 5 and 7 % of dry matter (DM) in a ration with a high roughage to concentrate ratio (70: 30). Total gas and CH4 were measured at 3, 6, 9, 12 and 24 h of fermentation while pH, volatile fatty acids (VFA), and ammonia nitrogen (NH3-N) concentrations were measured at 24 h using buffalo rumen fluid in an in vitro batch culture system. Microbial populations were determined using 16S-rDNA gene primers by RT-PCR. Results: The results revealed that linoleic acid at 3, 5 and 7 % decreased the concentration of NH3-N (p < 0.05) but linolenic acid at 5 and 7 % increased NH3-N (p < 0.05). A linear decrease (p <0.001) in acetate and butyrate, coupled with linear increase (p <0.001) in propionate was observed in response to treatment. Furthermore, supplementation of 3, 5 and 7 % of both fatty acids linearly (p < 0.001) decreased total gas and CH4 production when compared to the control. The addition of linoleic acid linearly (p < 0.001) decreased the number of protozoa without affecting methanogens, while linolenic acid linearly and quadratically (p < 0.001) reduced the population of both protozoa and methanogens (p < 0.05). Conclusion: Linolenic acid is more effective at a 3 % level in reducing methane production (up to 63 %) in high roughage diets. [ABSTRACT FROM AUTHOR]

This study was aimed to evaluate the potential of a herbal mixture (HM) to improve production performance, rumen fermentation, and milk fatty acid profile in water buffaloes. Sixteen Murrah buffaloes (in four groups) were fed for 10 weeks with the same basal diet supplemented with 0 (control); 20 (HM20), 30 (HM30), and 40 (HM40) g/buffalo per day. The herbal mixture contained an equal quantity of black pepper (fruit), ginger (tubers), cinnamon (bark), peppermint (leaves), ajwain (seeds) and garlic (bulbs). After two weeks of adaptation, daily milk yield, and weekly milk composition were recorded. On the last day of the experiment, rumen contents were collected to determine rumen fermentation parameters and bacterial diversity through 16S rRNA sequencing. Results revealed no effect of treatment on dry matter intake (DMI), rumen fermentation parameters, and daily milk yield. However, milk fat (%) showed a tendency to increase (p=0:07) in HM20 as compared with the control group. A significant increase in mono and polyunsaturated fatty acids (C14:1, C16:1, C18:2n6 and C18:3) whereas a decrease in saturated fatty acids (C18:0) in milk was observed in HM20 as compared with the control group. No significant change in bacterial diversity parameters (alpha and beta diversity) was observed in response to the treatment. Despite the substantial variation observed in the relative abundance of bacteria among treatment groups, no significant effect of treatment was observed when compared with the control group. Correlation analysis revealed several positive and negative correlations of rumen bacteria with rumen volatile fatty acids (VFA) and milk yield traits. Bacterial genera including Succinivibrionaceae, Butyrivibrio, Pseudobutyrivibrio, and Lachnospiraceae showed a positive correlation with VFA and milk yield traits. Overall, we observed 52 positive and 10 negative correlations of rumen bacteria with milk fatty acid contents. Our study revealed the potential of the herbal mixture at a lower supplemental level (20 g/day) to increase milk fat (%) and unsaturated fatty acid content in buffalo. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate potential of Moringa oleifera seed oil (MOSO) to modulate rumen microflora to mitigate methane (CH4) production in different total mixed rations (TMRs). Three TMRs with different roughage (R) to concentrate (C) ratio were used as substrates (R70:C30, R50:C50 and R30:C70) for in vitro fermentation study using batch culture technique. Results revealed that supplementation of MOSO in different rations with variable roughage to concentrates ratio altered (P < 0.05) CH4 production and fermentation parameters. M. oleifera seed oil at 3 and 4% increased (P < 0.01) microbial protein (MCP) and propionate concentration for all rations but decreased acetate concentration in R70:C30 TMR. Two levels of MOSO (3 and 4%) quadratically increased (P = 0.001) MCP and decreased acetate in R30:C70 while all levels increased propionate. For R30:C70 and R50:C50 rations, MOSO linearly and quadratically decreased (P < 0.001) protozoa and methanogen counts; however, protozoa, methanogens and bacteria were significantly increased in R70:C30 ration without any change in fungal counts. Supplementation of higher level of MOSO (4%) in high roughage ration (R70:C30) showed negative effects on diversity (Shannon index) and evenness of bacterial species as compared to control and lower oil level. Moreover, it also decreased Firmicutes to Bacteroidetes ratio in high roughage rations more obviously at lower levels. Moringa oil also stimulated Prevotella in both high and low roughages diets that indicates its potential to mediate rumen acidosis. Conclusively, MOSO enhanced fermentation kinetics and decreased CH4 production through effective modulation of rumen microbiome. [ABSTRACT FROM AUTHOR]