Both ruminal microbial structure and functionality might play a role in inter-individual variation in susceptibility for subacute rumen acidosis (SARA) observed in dairy cows. The aims of this study were to determine whether differences between cows with distinct SARA susceptibility were reflected in distinct (1) ruminal microbial communities, (2) salivary bacterial communities, and (3) fermentative capacity of ruminal microbiota assessed in vitro. To test this hypothesis, rumen samples were collected via an esophageal tube on 21 d postpartum from 38 multiparous Holstein cows, which were classified into 4 groups differing in median and mean time of reticular pH below 6 as well as area under the curve of pH below 6.0. During the 21 d postpartum, all cows within a group fulfilled following criteria: susceptible (S, n = 10; mean or median ≥180 min/d), moderately susceptible (MS, n = 7; 60 min/d < mean time of pH below 6 < 180 min/d, and median time of pH below 6 <180 min/d), moderately unsusceptible (MU, n = 11; 10 min/d < mean < 60 min/d, and median time of pH below 6 ≤30 min/d), or unsusceptible (U, n = 10; median = 0 min/d, and mean <10 min/d). Groups did not differ in total daily dry matter intake nor in total, roughage, or concentrate intake during daily 6-h time intervals. Rumen bacterial α-diversity did not differ among groups, but β-diversity varied and bacterial 16S rRNA gene copy numbers were lower in S compared with U cows. The relative abundance of genera Streptococcus, Sharpea, Prevotellaceae_YAB2003, Succinivibrionaceae_UCG-001, Ruminococcus, and Ruminococcaceae_UCG-001 were higher in S compared with U cows. In contrast, Lachnospiraceae_ND3007 and Oscillospiraceae_V9D2013 were more abundant in U cows. Although pH-associated, inter-animal differences were also observed in the salivary bacteria, common differences in ruminal and salivary bacterial genera were limited. The functionality of the rumen microbiota was evaluated in vitro through exposure of the microbial inoculum of S and U cows to an anaerobic buffer at pH 5.8 and 6.8, in the presence of sterile supernatant of their own and of dry cows' rumen fluid (2 × 2 design). Generally, the S inoculum produced more volatile fatty acids, except at low pH with dry cows' supernatant, where volatile fatty acid production was completely impaired and lactate accumulation was highest. Compared with the microbes of U cows, microbes of S cows showed less fermentative activity in situations with 2 stress factors (low pH and an unfamiliar environment, i.e., rumen fluid supernatant of dry cows).

The use of the fermented total mixed ration (FTMR) is a promising approach for the preservation of homogeneous feed, but changes during fermentation and links with the bacterial community of FTMR are not fully understood. This study investigated the effects of adding oat silage (OS) to the fermented total mixed ration (FTMR) in terms of fermentation, chemical composition, and the bacterial community. The fermentation quality of FTMR with 22% OS was greatly improved, as demonstrated by decreases in the butyric acid concentration, a lower lactic acid/acetic acid ratio, a larger population of lactic acid bacteria (LAB), and quicker spoilage yeast death. Further examination of the effects of various ensiling days on nutritive values showed stable crude protein and nonprotein nitrogen (NPN) contents. The concentrations of acetic acid, propionic acid, and ammonia–nitrogen (NH₃–N) were increased following all FTMR treatments after 15 d, while the concentration of water-soluble carbohydrates (WSC) was decreased. More heterofermentative LAB, such as Lentilactobacillus buchneri, Lentilactobacillus brevis, and Companilactobacillus versmoldensis were found after adding 11% and 22% OS. Moreover, the addition of 22% OS caused a marked increase in both bacterial richness and diversity, dominated by the Lactobacillus genus complex. Among species of the Lactobacillus genus complex, the occurrence of Loigolactobacillus coryniformis was positively correlated with lactic acid, NPN, and NH₃–N concentrations, suggesting its potential role in altering the fermentation profiles.

The main objective of this study was to establish the metabolic pathway of a biogas fermentation system active at low temperature, and to describe the bacterial and archaeal methanogenic species responsible. A biogas fermentation inoculum adapted to be active at 9°C was used in a 10‐L batch‐type temperature‐controlled fermentor, using pig manure as the raw material. Fermentation at 9°C was followed for 120 days. A combination of nonbiological and biological factors was analyzed. Species were identified from OTU analysis based on 16S rDNA amplicon sequencing, and correlation analysis was used to study the effect of nonbiological factors, bacterial communities, and archaea communities. Biogas production was most effective between days 50 and 90, where gas production was above 0.70 L/d and the CH₄ yield was above 0.36 L/g‐volatile solid. The relative abundance of the first dominant bacterial OTU with 99% identity to Clostridium cellulovorans varied between 16.17% and 27.30%. These bacteria typically degrade cellulose and hemicellulose. The second dominant bacterial OTU (relative abundance 15.23%‐27.15%) was 99% identical to Terrisporobacter petrolearius, a typical fermentative species. The most abundant (4.15%‐37.14%) archaeal OTU was 98% identical to Methanocorpusculum sinense, which is a typical hydrogenotrophic methanogen. A low‐temperature biogas metabolic pathway was constructed, based on abundant bacterial and methanogen OTUs. The most suitable metabolic pathway describes that hydrogen is produced by one part of the community and then used to reduce the produced carbon dioxide into CH₄; this seems to be the major pathway by which biogas is produced under low‐temperature conditions.

To improve the low energy production efficiency of fermentative hydrogen production (FHP) and fermentative methane production (FMP), the two types of fermentation were effectively combined in this study. Using sucrose as feedstock, both successive fermentative production of hydrogen followed by methane (SFPHM) and successive fermentative production of methane followed by hydrogen (SFPMH) were performed to improve the energy production efficiency of the overall anaerobic process. The results showed that the energy conversion efficiency of SFPHM was 79.85% whereas that of SFPMH was only 43.12%. Therefore, SFPHM can significantly improve the energy production efficiency of traditional anaerobic biological treatment.