This study evaluated the effects of different dietary n-6/n-3 polyunsaturated fatty acid (PUFA) ratios on in vitro ruminant fermentation. Methane production, fatty acid composition, and microbial profiles were compared after the in vitro fermentation of rumen fluid collected from cows that had been fed isoenergetic and isoproteic experimental diets at three different n-6/n-3 ratios: 3.04 (HN6, high n-6 source), 2.03 (MN6, medium n-6 source), and 0.8 (LN6, low n-6 source). The fermented rumen fluid pH and total volatile fatty acid (VFA) levels were significantly decreased (p < 0.05) in the HN6 group as compared with those in the MN6 and LN6 groups. Additionally, the HN6 group produced a significantly lower (p < 0.05) proportion of methane than the MN6 group during in vitro fermentation. The MN6 and LN6 groups had significantly increased (p < 0.05) levels of C18:2n6 and C18:3n3 in the fermented rumen fluid, respectively, as compared with the HN6 group. The Chao 1 diversity index value was lower (p < 0.05) in the HN6 group than in the MN6 and LN6 groups. The observed species richness was significantly lower (p < 0.05) in the HN6 group than in the MN6 group. The reduced relative abundances of Lachnospiraceae UCG-006 and Selenomonas in the HN6 group resulted in lower pH and VFA levels (i.e., acetate, propionate, butyrate, and total VFA) during in vitro fermentation. Furthermore, n-6 and n-3 PUFAs were toxic to Butyrivibrio_2 growth, resulting in high levels of incomplete biohydrogenation. Taken together, the study findings suggest that supplementation of high-forage diets with high levels of n-6 PUFAs could reduce methane emissions, whereas both VFA concentration and pH are reduced.

This study aimed to investigate whether agricultural by-products with a high NDF content and small-particle-size substitute for forage could cause hindgut acidosis and dysbacteriosis in lactating dairy cows. We investigated the impact of soybean hull and beet pulp on the fecal fermentation, bacterial community, and digestibility of cows. Sixteen lactating Holstein cows were treated as follows (% of dry matter (DM)): amount of by-product added was 0 (control, CON), 1.67% (low by-products, LB), 3.33% (medium by-products, MB), and 5% (high by-products, HB). The results showed the fecal pH of cows to be 7.23–7.29, implying no hindgut acidosis. With increased inclusion of by-products in the diets, the proportion of fecal propionate; relative abundance of the phylum Bacteroidetes, the family Lachnospiraceae, and genera unclassified_f_Lachnospiraceae, Acetitomaculum, and Prevotella; and the DM and NDF digestibility of cows all increased linearly. Meanwhile, the fecal genera Turicibacter and Clostridium_sensu_stricto_1 decreased linearly. By-products promoted the abundance of fecal bacteria genes related to energy metabolism, glycolysis/gluconeogenesis, and propanoate metabolism; and correlations between fecal short chain fatty acids, digestibility, and the bacteria genera were seen. Overall, our study suggested that adding 5% by-products could be a viable dietary formulation strategy that promotes digestibility and makes positive changes in hindgut fermentation and bacteria.

It is well known that dairy cows are fed diets with high fat content, which can adversely affect rumen fermentation. However, whether the effects of high fat content on rumen fermentation are related to the composition of fatty acids (FA) is for further study. We explored the effects of unsaturated fatty acid (UFA) ratios in vitro on rumen, methane concentration and microbial composition under the same fat levels. The experiment included a low-unsaturated group (LU, UFA proportion: 42.8%), a medium-unsaturated group (MU, UFA proportion: 56.9%), and a high-unsaturated group (HU, UFA proportion: 70.9%). The incubation fluid pH and NH₃-N levels were not significantly different in the three groups. Total volatile fatty acid (TVFA), acetate, propionate, butyrate, and valerate in the MU group had a decreased trend compared to the LU group (0.05 < p < 0.1), and no difference was found in other volatile fatty acids (VFAs) among the three groups. Furthermore, gas production kinetic parameters among the three groups did not differ significantly. The LU group’s CH₄ concentration was significantly higher than the HU group (p < 0.05). The CO₂ concentration in the LU group was also significantly higher than the MU and LU groups (p < 0.05). Additionally, 16S rRNA microbial sequencing results showed that the Shannon diversity value significantly increased in the MU group (p < 0.05) compared to the LU group. Other alpha diversity indices (Chao 1, observed species, and ACE) did not differ among the three groups. The increased proportion of UFA significantly decreased the relative abundance of Succinivibrionaceae_UCG_001 and Fibrobacter (p < 0.05). Meanwhile, the multiple Lachnospiraceae bacteria significantly increased in the MU group (p < 0.05). Overall, our findings indicated that the microbial community in the incubation system could be affected by elevating proportions of UFA, affecting the yield of VFA, whereas the CH₄ concentration was reduced.

Lipid metabolism plays an important role in the energy economy of ruminants. However, its interactions of fat, rumen fermentation, gas emission, and microorganisms are not yet clear. This study evaluated the effect of adding raw oilseeds to high-forage diets on in vitro ruminal fermentation, gas composition, and microbial profile. Three isoenergetic and isoproteic experimental diets were designed and used as fermentation substrate: control treatment (CON group) was the basal diet lacking oilseeds, the other two treatments were the basal diet supplemented by 100 g/kg dry matter (DM) raw whole soybean (S group) and 50 g/kg DM raw flaxseed (F group), respectively. Data showed that the acetate, butyrate, and total VFA concentration of culture fluids in the S group were lower (p < 0.05) than in the F group. There was a tendency to a higher level (p = 0.094) of propionate concentration in the F group compared with the other two groups. The gas production in the F group was higher (p < 0.05) than in the control group. There was a lower abundance of Sutterella (p < 0.05) and a greater abundance of Butyrivibrio (p < 0.05) in both of the two oilseed treatments. Methanobrevibacter (p = 0.078) in the F group was the lowest. Our results suggested that CH₄ emission could be inhibited with flaxseed supplementation by propionate production metabolism, biohydrogenation of unsaturated fatty acid (FA), and toxicity to Methanobrevibacter, while regarding soybean seed supplementation, the emission of CH₄ was more likely to be reduced through biohydrogenation of unsaturated FA modulated by Butyrivibrio.

This study was conducted to investigate the effect of three rates of prepartum dietary concentrate feeding on the milk performance, energy balance, and rumen fermentation of Montbéliarde–Holstein crossbred cows. Eighteen transition Montbéliarde–Holstein crossbred cows with similar days of gestation (258 ± 12 day) and body weights (622 ± 44 kg) were selected and randomly divided into three groups. In the prepartum period, the addition of concentrates accounted for 0.3% (low concentrate, LC), 0.6% (medium concentrate, MC), and 0.9% (high concentrate, HC) of the cow’s body weight. The forage was corn stover, which was fed to the cows ad libitum with free access to water. Postpartum, all of the cows were fed a common lactation total mixed ration. The experimental period lasted from 21 days prepartum to 28 days postpartum. The energy balance (EB), net energy intake (NEI), and dry matter intake (DMI) of the HC group were greater than those of the other groups (p < 0.05). Likewise, the non-esterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), and total bilirubin (TBIL) in the blood of the LC group had significantly higher concentrations than they did in the other groups (p < 0.05). Moreover, the increase in the level of dietary concentrate had no significant effect on the rumen fermentation parameters (p > 0.05), and the total intestinal digestibility of dry matter (DM), crude protein (CP), and ether extract (EE) in the HC group was significantly higher (p < 0.05) than it was in the other groups during the prepartum period. In conclusion, the administration of the MC diet in the prepartum period had no negative effect on the performance and rumen fermentation of postpartum dairy cows and can satisfy the energy needs of prepartum dairy cows. Therefore, under our experimental conditions, the 0.6% prenatal concentrate feeding amount was the most appropriate for Montbéliarde–Holstein crossbred cows.