Subacute ruminal acidosis (SARA) negatively impacts the dairy industry by decreasing dry matter intake, milk production, profitability, and increasing culling rate and death loss. Six ruminally cannulated, lactating Holstein cows were used in a replicated incomplete Latin square design to determine the effects of SARA induction on the ruminal microbiome and epithelium. Experimental periods were 10 days with days 1–3 for ad libitum intake of control diet, followed by 50% feed restriction on day 4, and ad libitum access on day 5 to the basal diet or the basal diet with an additional 10% of a 50:50 wheat/barley pellet. Based on subsequent ruminal pH, cows were grouped (SARA grouping; SG) as Non-SARA or SARA based on time <5.6 pH (0 and 3.4 h, respectively). Ruminal samples were collected on days 1 and 6 of each period prior to feeding and separated into liquid and solid fractions. Microbial DNA was extracted for bacterial analysis using 16S rRNA gene paired-end sequencing on the MiSeq Illumina platform and quantitative PCR (qPCR). Ruminal epithelium biopsies were taken on days 1 and 6 before feeding. Quantitative RT-PCR was used to determine gene expression in rumen epithelium. Bray–Curtis similarity indicated samples within the liquid fraction separated by day and coincided with an increased relative abundance of genera Prevotella, Ruminococcus, Streptococcus, and Lactobacillus on day 6 (P < 0.06). Although Firmicutes was the predominant phyla in the solid fraction, a SG × day interaction (P < 0.01) indicated a decrease on day 6 for SARA cows. In contrast, phylum Bacteroidetes increased on day 6 (P < 0.01) for SARA cows driven by greater genera Prevotella and YRC22 (P < 0.01). Streptococcus bovis and Succinivibrio dextrinosolvens populations tended to increase on day 6 but were not affected by SG. In ruminal epithelium, CLDN1 and CLDN4 expression increased on day 6 (P < 0.03) 24 h after SARA induction and a tendency for a SG × day interaction (P < 0.10) was observed for CLDN4. Overall, results indicate more rapid adaptation to an induced bout of SARA in the solid fraction ruminal microbiome compared with ruminal epithelium.

The usage of direct-fed microbials (DFM) has become common in the dairy industry, but questions regarding choice of strain, mode of action, and efficacy remain prevalent. The objective of this study was to evaluate the effects of a DFM (Bacillus pumilus 8G-134) on pre- and postpartum performance and incidence of subclinical ketosis in early lactation. Forty-three multiparous Holstein cows were assigned to 2 treatments in a randomized complete block design; cows in the direct-fed microbial treatment (DFMt, n=21) received 5.0×10(9) cfu/cow of B. pumilus in 28 g of a maltodextrin carrier, whereas cows in the control treatment (CON, n=22) received 28 g of maltodextrin carrier alone. Treatments were top-dressed on the total mixed ration daily. Treatments were applied from 21 d before expected calving date to 154 d after calving. Cows on treatment DFMt tended to have lower serum haptoglobin concentration than CON cows on d 14. Cows on treatment DFMt had higher IgA concentrations in milk than CON cows during the first week after calving. Cows fed DFMt had higher yields of milk, fat-corrected milk, energy-corrected milk, milk fat, and milk protein during the second week of lactation than CON; however, we found no differences between treatments on milk yield and milk components overall. Cows on DFMt tended to have higher feed conversion and to have lower prevalence of subclinical ketosis (beta-hydroxybutyrate >1.2 mmol/L) on d 5 than cows fed CON. Dry matter intake, body weight, and body condition score were not affected by DFMt supplementation. Milk production efficiencies (calculated based on fat-corrected milk and energy-corrected milk) were higher by 0.1 kg of milk per kilogram of dry matter intake in cows that received DFMt compared with cows that received CON. In conclusion, cows receiving DFMt tended to have lower incidence of subclinical ketosis than cows receiving CON. Cows fed DFMt tended to have higher feed conversion and evidence for greater immunity than CON. Supplementation with B pumilus 8G-134 may provide benefits for transition cow health and milk production efficiency.

The effects of a grain challenge on ruminal, urine, and fecal pH, apparent total-tract starch digestibility, and milk composition were determined. Six Holstein cows, 6 rumen-cannulated Holstein cows, and 6 Jersey cows were used in a replicated 3 × 3 Latin square design balanced to measure carryover effects. Periods (10 d) were divided into 4 stages (S): S1, d 1 to 3, served as baseline with regular total mixed ration ad libitum; S2, d 4, served as restricted feeding, with cows offered 50% of the total mixed ration fed on S1 (dry matter basis); S3, d 5, a grain challenge was performed, in which cows were fed total mixed ration ad libitum and not fed (CON) or fed an addition of 10% (MG) or 20% (HG) pellet wheat-barley (1:1) top-dressed onto the total mixed ration, based on dry matter intake obtained in S1; S4, d 6 to 10, served as recovery stage with regular total mixed ration fed ad libitum. Overall, cows had a quadratic treatment effect for milk yield where CON (22.6 kg/d) and HG (23.5 kg/d) had lower milk yield than cows in MG (23.7 kg/d). Jersey cows had a quadratic treatment effect for dry matter intake where cows in CON (13.2 kg/d) and HG (12.4 kg/d) had lower dry matter intake than cows in MG (14 kg/d). Holstein cows had a linear treatment effect for dry matter intake (17.7, 18.4, and 18.6 kg/d for CON, MG, and HG, respectively). Rumen pH for the rumen-cannulated cows had a linear treatment effect (6.45, 6.35, and 6.24 for CON, MG, and HG, respectively). Cows in HG spent more time with rumen pH below 5.8 (4.33 h) than MG (2 h) or CON (2.17 h) as shown by the quadratic treatment effect. Holstein cows in HG (8.46) had lower urine pH than MG (8.51) or CON (8.54) as showed by the linear treatment effect for urine pH. Apparent total-tract starch digestibility had a tendency for a linear treatment effect on S3 (97.62 ± 1.5, 97.47 ± 1.5, and 91.84 ± 1.6%, for CON, MG, and HG, respectively). Fecal pH was associated with rumen pH depression as early as 15 h after feeding for Holstein cows. In conclusion, a grain challenge reduced urine pH in Holstein cows but not in Jersey cows. Holstein cows' health were not affected when rumen pH was depressed. A potentially useful link between rumen pH and systemic (urine) pH within 2 h after feeding was quantified in Holstein cows.

The objective of the present study was to investigate the effects of virginiamycin (VM ) supplementation on ruminal fermentation and microbial populations in steers. Four ruminally cannulated Chinese Luxi steers (BW 559.4 &plusmn; 30.1 kg) were used in a crossover design experiment with an experimental period of 28 days. The forage to concentrate ratio of the basal diet was 35:65 on dry matter basis. The experiment consisted of control treatment and treatment with control diet plus VM at a dose of 30 mg/kg concentrate (DM basis). Rumen fluid was collected at 07:30 prefeeding, at 11:30 and 17:30 postfeeding on day 27 and 28. A part of the pooled sample from rumen fluid was transferred to anaerobic culture by a roll-tube technique and analysed for species-specific real-time PCR quantification. The remaining pooled rumen fluid sample was analyzed for pH, VFA, ammonia N and l-lactic acid. The results showed that VM increased the ruminal pH (6.70 vs. 6.63; P &lt; 0.05), but it decreased ammonia nitrogen (4.94 vs. 6.19 mg/100 ml; P &lt; 0.01) and mean counts of amylolytic bacteria and proteolytic bacteria (P &lt; 0.01) as compared to the control. The additive VM did not affect the l-lactic acid concentration (1.39 vs. 1.26 mmol/l) in rumen fluid. Compared to the control, the steers receiving VM have altered a trend of quantification of Selenomonas ruminantium, Anaerovibrio lipolytica, Ruminococcus albus and Streptococcus bovis in rumen fluid (0.05&lt;p&lt;0.1) as compared to the control. However, VM had no significant effect on Lactobacillus spp. (P = 0.41), Butyrivibrio fibrisolvens (P = 0.35), on the genus Ruminococcus (P = 0.25), Ruminococcus flavefaciens (P = 0.52), Prevotella ruminicola (P = 0.54), on the genus Prevotella (P = 0.67) and Megasphaera elsdenii (P = 0.97). In this study, we found that VM had selective effects on ruminal bacteria and influenced ruminal fermentation by changing a part of the specific ruminal bacteria populations.