fixate the cells and bacteria for 30 minutes and then washed with pbs three times finally 100 u03bcl crystal violet was added for staining and was quickly rinsed off with a small amount of 75 alcohol to remove the dye on cells after 5 minutes d phase contrast microscope was used to observe the adherence of lactobacilli to intestinal epithelial cells and the number of adherent lactobacilli per 50 cells was counted in randomized microscopic fields finally the average adherent lactobacilli per cell were calculated in vivo animal testing was carried out using a modified version of usman and hosono 2000 syrian male hamsters used for the tests were purchased from the national laboratory animal breeding amp research center in taiwan the hamsters were 7 u02dc 8 weeks old and weighed 82 98 grams averaging 95 3 grams before the test each hamster was separately bred in individual ventilated cage ivc under 25 u00b0 c and the light dark cycle of 12 hours in light and 12 hours in darkness feeds and sterile ro water were provided ad libitum during the one week acclimatization period the hamsters were on ain 93 diet but feeds with different proportions of ingredients were provided after the hamsters were grouped.