The aim of this study was to investigate the effect of high temperature on the viability of probiotic organisms (Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae) mixed with animal feed under controlled conditions by simulating a farm feed bin in the summer. Following inoculation of probiotics into the feed, the pH and probiotic viability were monitored during an 8-day incubation at room temperature. Sterile and non-sterile feeds displayed different patterns of pH changes, with increased pH in non-sterile feed at 2 days, but a pattern of decreasing pH at 4 days. The viabilities of S. cerevisiae and B. subtilis after mono/co-inoculation were maintained without substantial changes during the incubation, whereas L. plantarum viability tended to decline. In both non-sterile and sterile feeds, the probiotics were maintained or grew without any antagonistic effects. Probiotic viability was also tested upon a shift to high temperature (60°C). There was no distinct change in pH between sterile and non-sterile feeds after the temperature shift. L. plantarum and S. cerevisiae could not survive at the high temperature, whereas B. subtilis displayed normal growth, and it inhibited the growth of contaminant microbes. Fungal growth was not observed in non-sterile feed 2 days after supplementation with B. subtilis. Therefore, heat resistant B. subtilis could be safely used in feed bins to inhibit microbial contamination, even at high temperatures. The prevention of elevated temperature in feed bins is necessary for the utilization of L. plantarum and S. cerevisiae during the summer season.
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