Preservation of gastrointestinal bacteria and their microenvironmental associations in rats by freezing

The use of frozen rat gastrointestinal tissue samples for both the recovery of viable bacteria and for observation of microbial communities associated with the tissue was investigated. A decrease of 1 log in lactobacilli, bifidobacteria, and anaerobes was observed when the numbers of bacteria recoverable from frozen tissue (stored 7 to 9 days) were compared to those recoverable from fresh nonfrozen tissue (zero time control). However, freezing did not appear to decrease the numbers of recoverable coliforms. Tissues, cleaved with razor blades after being frozen and stored for 7 to 9 days, showed bacterial communities situated on the mucosa and in the lumen of gastrointestinal specimens. This freezing technique preserved structures not previously observed in the gastrointestinal tract. This indicates that freezing is a good method to use to study such fragile microenvironments.