Evaluation of the microbial population in ruminal fluid using real time PCR in steers treated with virginiamycin

Abstract

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 ± 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 < 0.05), but it decreased ammonia nitrogen (4.94 vs. 6.19 mg/100 ml; P < 0.01) and mean counts of amylolytic bacteria and proteolytic bacteria (P < 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<p<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.

Keywords

• virginiamycin
• roll-tube technique
• ruminal fermentation parameters
• real-time pcr