We investigated the relationship between ruminal pH and bacteria in calves fed calf starter with and without forage during weaning transition. First, 16 Holstein bull calves were obtained from dairy farms and equipped with rumen cannulas by cannulation surgery. Then, calves (73.5 ± 4.2 kg; mean ± SE) were assigned to groups fed calf starter either with forage (HAY, n = 8) or without forage (CON, n = 8), and all calves were weaned at 8 weeks of age. Ruminal pH was measured continuously, and rumen fluid samples were collected at 7, 8, 9, and 11 weeks of age, namely −1, 0, 1, and 3 weeks after weaning, respectively, to assess volatile fatty acid concentrations and bacterial DNA. The 24-h mean ruminal pH was significantly (P < 0.05) different between the two groups. Diurnal changes in the 1-h mean ruminal pH were observed throughout the study in the HAY group; however, they were not observed at 0 and 1 weeks after weaning in the CON group. Moreover, the HAY group had significantly (P < 0.05) higher proportions of acetate and butyrate and lower proportion of propionate, and significantly (P < 0.05) lower ruminal acetate-to-propionate ratios were observed in the CON group. The ruminal bacterial diversity indices decreased after −1 week in both groups and increased at 0 and 1 weeks after weaning in the HAY and CON groups, respectively. From the 454 pyrosequencing analysis, significant differences (P < 0.05) were observed in the relative abundance of several phyla (Bacteroidetes, Actinobacteria, and Tenericutes) and one genus (Prevotella) between the two groups. From quantitative real-time PCR analysis, the HAY group had the higher copy numbers of cellulolytic bacteria (Ruminococcus flavefaciens and Ruminococcus albus) compared with the CON group. This study demonstrated that feeding of dietary forage alleviates subacute ruminal acidosis due to diurnal changes in ruminal pH. Furthermore, changes in ruminal pH affect the ruminal bacterial diversity and relative abundance, and these changes might have influenced the establishment of fermentative ruminal functions during weaning transition.

We investigated the effect of repeated subacute ruminal acidosis (SARA) challenges on the pH, fermentative function, and bacterial community in the rumen. Four rumen-cannulated Holstein bulls were fed a high-forage diet for 7 d (HF period) followed by a high-grain diet for 7 d (HG period). Four SARA challenges were carried out consecutively (first, second, third, and fourth challenges). The ruminal pH was measured continuously during the experiment, and rumen fluid samples during the first to fourth challenges were collected at 0800, 1400, and 2000 h on the last days of each feeding period for analysis; volatile fatty acid components, NH-N, and lactic acid concentrations were measured. Bacterial community structure was analyzed at 0800 h during the first and fourth challenges on the last days of each period. The 24-h mean ruminal pH was decreased during the transition from high-forage to high-grain diet and tended to differ between the HF and HG periods. During the HG period, ruminal pH <5.6 was maintained for a longer period in the first and second challenges (350 and 405 min/d, respectively) than in the third and fourth challenges (both 120 min/d). A marked increase in total volatile fatty acid and NH-N concentrations during the HG period was observed in the later challenges. In addition, lower and higher proportions of acetic and butyric acids, respectively, were observed during the HG period than during the HF period. A total of 37 core bacterial genera were found in all samples; however, the relative abundance of several genera differed significantly between the HF and HG periods (Prevotella, Ruminococcus, Eubacterium, and Oscillibacter) and between the first and fourth challenges (Eubacterium and unclassified Clostridiaceae). During the HG period, lower relative abundances of Prevotella, Eubacterium, and Oscillibacter and higher relative abundance of Ruminococcus were detected compared with during the HF period. The relative abundances of Eubacterium and unclassified Clostridiaceae were lower in the first challenge than in the fourth challenge. Bacterial diversity was greater during the HF period than during the HG period and was greater during the fourth challenge than during the first challenge. Interestingly, diversity indices during the HG period of the fourth challenge were higher than those during the HF period of the first challenge. These results suggested that rumen fermentation in Holstein cattle can adapt to repeated SARA challenges by minimizing the adverse changes in ruminal pH. Moreover, the composition and diversity of the ruminal bacterial community may be affected by ruminal pH and vice versa.

BackgroundSubacute ruminal acidosis (SARA) is characterized by a ruminal pH depression, and microbiota can also be affected by a higher acidity and/or dietary changes. Previous studies have revealed similar patterns in pH reduction in the rumen and reticulum, whereas changes in reticular pH and bacterial community following a high-grain diet are not fully understood. The aim of this study was to investigate the changes in reticular pH and bacterial community structure following a high-grain diet simultaneously with those in the rumen.ResultsSARA was diagnosed when ruminal and reticular pH remained under 5.6 for 350 ± 14 and 312 ± 24 min/d, respectively, on the last day of the CON period. During the CON period, lower proportion of acetic acid and higher proportion of butyric acid were observed compared with the HAY period. The proportions of acetic acid and propionic acid were lower and higher, respectively, in the rumen compared with the reticulum. From 454 pyrosequencing analysis, the relative abundance of several genera differed significantly between the two periods and the two locations. During the HAY period, higher relative abundances of Prevotella, Eubacterium, Oscillibacter, and Succiniclasticum and lower relative abundances of Ruminococcus, Clostridium, and Olsenella were identified compared with the CON period. Furthermore, the relative abundance of Eubacterium was lower in the rumen compared with the reticulum. Bacterial diversity indices were significantly different between the HAY and CON periods, being higher in the HAY period. The quantitative real-time PCR showed that the copy numbers of several cellulolytic bacteria (Fibrobacter succinogenes and Ruminococcus albus) were higher during the HAY period.ConclusionA high-grain diet showed similar impacts on the pH, fermentation, and bacterial community structure in the rumen and reticulum. During the CON period, ruminal and reticular pH decreased following the high-grain challenge, and lower bacterial diversity and changes in the bacterial composition, similarity, and bacterial copy numbers were observed due to a higher acidity and dietary changes compared with the HAY period. These changes may influence the fermentative ability of the rumen and reticulum.Electronic supplementary materialThe online version of this article (10.1186/s12917-018-1637-3) contains supplementary material, which is available to authorized users.

We investigated the relationship between ruminal pH and transcriptomic adaptation of the rumen epithelium (RE) of calves fed calf starter with and without forage during the weaning transition. Holstein calves were assigned to groups fed calf starter either with forage (HAY group, n = 3) or without forage (CON group, n = 4). Ruminal pH was measured continuously, and rumen fluid and epithelium were collected 3 wk after weaning. mRNA expression profiles of the RE were examined by one-color microarray. Differentially expressed genes (DEGs) were investigated using the Ingenuity Pathway Analysis (IPA). Mean and maximum ruminal pH were significantly (P < 0.05) higher, and the duration of pH < 5.8 during 1 day was significantly (P < 0.05) shorter, in the HAY group. The proportion of ruminal acetate and the acetate-to-propionate ratio were significantly (P < 0.05) lower in the CON group. DEGs encoding transcription regulators (SREBP1), insulin-like growth factor binding proteins (IGFBP7 and CTGF), ketogenic enzymes (HMGCL, BDH1, and BDH2), and a transporter (SLC16A3) were identified (P < 0.05) between the two groups. A growth factor (TGFB1) and signaling pathway (EGF and EGFR) were activated as upstream regulators. These results suggest that dietary forage alleviates ruminal acidosis, and the decrease in ruminal pH may damage the RE, leading to changes in gene expression to repair the damage. Furthermore, rumen development may be regulated by growth factor (TGFB1) and signaling pathways (EGF and IGFBP) for adaptation to feeding on calf starter with and without forage during the weaning transition.

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