Consistent with the 25% forage and 10% protein diet that these cattle were being fed, the RF comprised a higher percentage of acetate [28–31]. Acetate ranged from 72-62%, compared to the 13-18% propionate and 6-13% butyrate concentrations across the uRF, dRF and fRF samples in both experiments, irrespective of procedures used to prepare dRF and fRF (Tables 1 and 2). LB broth (pH 7.0-7.2) did not contain added VFAs. O157 growth characteristics Log phase O157 cultures, set up for the two experiments, were at 0.5-0.6 OD600, respectively, with viable counts around 1 × 108 cfu/ml. Hence, when each medium was inoculated selleck chemicals to a starting 0.05-0.06 OD600, the corresponding O157 counts were at ~1-5
× 107 cfu/ml. In both experiments, O157 grew to an OD600 of 1.0 within
2 h in LB media, aerobically and anaerobically as anticipated, Everolimus concentration with an increase in viable count to 4 × 108 cfu/ml and the final culture pH at 6.0-6.2. However, significant differences were observed between aerobic and anaerobic growth patterns of O157 when cultured in dRF, fRF and uRF preparations. In Experiment I, O157 cultured in dRF and fRF achieved an average OD600 of 0.6-1.0 in 48 h aerobically, but remained at a low OD600 of ≤0.2 anaerobically, even after 14 days of incubation. Irrespective of the ODs, viable O157 was recovered from all cultures, but the viable counts at 106 (dRF)-2 × 107 (fRF) cfu/ml aerobically, and at 105 (dRF)-2 × 105 (fRF) Palbociclib chemical structure cfu/ml anaerobically (data not shown) appeared to be static or decreasing. The pH for dRF and fRF cultures at the end of incubation was around 7.7 (aerobic)–7.3 (anaerobic). Similar O157 growth results were observed upon anaerobic culture for 48 h in dRF, fRF and uRF, in Experiment II (Figure 1), with the pH for uRF cultures being
6.8 at end of incubation. This was despite these media being prepared with RF from a separate animal and a shorter anaerobic incubation period than in the first experiment, thereby verifying the observations made initially. Here, the cultures reached an average OD600 of 0.97 (LB), ~0.03 (dRF), ~0.04 (fRF) and ~0.03 (uRF) in 48 h, with O157 viable counts of 2 × 108 cfu/ml (LB), 4 × 105 cfu/ml (dRF), 3 × 106 cfu/ml (fRF) and 1 × 106 cfu/ml (uRF), respectively. Figure 1 Growth characteristics of O157 in Experiment II, following anaerobic incubation for 48 h, in LB and RF-preparations. Optical densities (OD600) and viable counts (colony forming units [cfu]/ml), with the standard error of means, are shown in graph A and B, respectively. The p values shown on the graphs were calculated using the Student t-Test (significant, p < 0.05). Significant differences were observed among the optical densities and viable counts of LB cultures versus RF-preparation cultures, under all growth conditions. However, differences between the PLX3397 ic50 RF-preparations were not always significant (Figure 1).