Time of rumen fluid collection relative to feeding alters in vitro fermentation volatile fatty acid production

The objective of this study was to determine the effect of time of rumen fluid collection relative to feeding on volatile fatty acid (VFA) production for in vitro rumen fermentation. Three ruminally cannulated Holstein heifers were used as rumen fluid donors. Feed was removed from heifers 12 h prior to feeding, rumen fluid was collected from each heifer before feeding (0 h), and at 2, 4, and 6 h after feeding, repeated on three separate incubation days. Buffered rumen fluid (100 mL) was incubated in 250-mL bottles containing 1.4 g of dried TMR, in duplicate for each heifer at each collection time. All bottles were incubated for 24 h at 39°C and constant agitation (60 rpm), and capped with monitors to capture temperature and pressure every 15 min (RF1, Ankom Technology, Macedon, NY). At the end of incubation, final pH and a sample of rumen fluid were collected for VFA and ammonia nitrogen. Data were analyzed using PROC GLIMMIX of SAS, with donor as the experimental unit and day as the random blocking factor; significance is defined as P ≤ 0.05. Time of rumen fluid collection significantly affected acetate (mmol/liter; P = 0.0004), propionate (mmol/liter; P = 0.02), isobutyrate (mmol/liter; P < 0.0001), valerate (mmol/liter; P = 0.004), isovalerate (mmol/liter; P < 0.00001), and total VFA concentrations (mmol/liter; P = 0.004). All VFA relative proportions were altered due to time of rumen fluid collection (P < 0.02). VFA production was highest when rumen fluid was collected 4-h post-feeding. There was little to no effect on pH. Our findings suggest that VFA production is maximized when rumen fluid is collected between 2 and 4 h after feeding.