Oral-stomach sampling to replace rumen-fistulated animals in ruminant nutrition research - a case study
Studies using fistulated ruminants have shed light on digestion and fermentation in the rumen and yielded tools to determine the feed efficiency of diets, limit effluents or anticipate variability in the quality of animal products. However, the ethical acceptability of such studies has been called into question. Our objective was to determine whether oral-stomach sampling (OSS) is an acceptable alternative to sampling through a ruminal cannula in characterizing the variability of rumen fluid composition induced by an acidogenic dietary challenge in dairy cows. During three four-week periods (P1, P2, P3), six rumen-fistulated cows were fed a standard diet based on supplemented corn silage (periods 1 and 3) alternating with a diet enriched with starch (period 2). Rumen juice was collected through cannulas at three locations in the rumen (reticulum, ventral sac or a mix of both) and by OSS, once per week at 0830 h, i.e. before morning feeding, and once every third week of each period at 1330 h, i.e. 4.5 hours after morning feeding. Whatever the sampling method or location, ruminal pH was lower in period 2 compared to periods 1 and 3 at 1330 h (6.21 vs. 5.57 vs. 6.12 in periods 1, 2 and 3). Ruminal pH was higher when obtained by OSS rather than cannulation, whatever the sampling location (on average, +0.44 points at 0830 h and +0.56 points at 1330 h). Mineral composition indicated a presumed dilution by saliva of OSS samples. This was also consistent with lower concentrations of volatile fatty acids. The next steps will be to analyze the associated variations in in vitro rumen fermentation parameters and microbiota composition. ImplicationsThe use of fistulated ruminants to elucidate ruminal physiology is currently controversial in society. In order to minimise, reduce or replace these experimental models, alternatives to ruminal fistula sampling are being evaluated. Here we focus on oral stomach sampling (OSS). We show that OSS can be a satisfactory alternative to cannula sampling in the evaluation of rumen fermentation parameters, especially for the molar proportion of volatile fatty acids, rumen pH, and rumen volatile fatty acid and ammonia concentrations. We also show that OSS must be standardized to avoid salivary contamination affecting the quality of the results.