72 preliminary data on the presence of bacteria in the uterus of pregnant cows

Bacterial invasion of the uterus during the postpartum period has been well described. Recent papers using 16S rRNA gene sequencing techniques suggest that the nonpregnant uterus contains a diverse flora of bacteria that are not necessarily pathogenic. In contrast, the pregnant uterus has until now been considered a sterile environment. The aim of the present study was to investigate whether bacteria were present in the uteri of pregnant cows. Uteri from pregnant, slaughtered animals (n=47) were sampled. The surface of the uterus was wiped with alcohol, flame sterilized, and cut open with sterile scissors. Samples were taken from the endometrium and from the placentomes. The samples were embedded in paraffin, sectioned at 3 microns, and prepared for fluorescence in situ hybridization using a probe targeting the 16S rRNA of the domain bacteria, so that all bacteria regardless of species were visualised. Using fluorescence microscopy, the presence of bacteria within or on the surface of the endometrium and within the placentomes was noted. The stage of pregnancy was estimated to range from 26 to 263 days by measuring the size of the embryo or fetus. The endometrial samples from 85.1% (40/47) of pregnant cows contained bacteria. In 22 cows, the bacteria were localised within the endometrial tissue, whereas in the remaining 18 cows, the bacteria were on the epithelial surface. Placental samples were obtained from 43 cows, and 76.7% (33/43) of these contained bacteria. The presence of bacteria in the pregnant uterus may suggest that a cow can carry a pregnancy despite the presence of few potentially pathogenic bacteria or that normal flora exist in the uterus as in, for example, the vagina. In conclusion, bacteria were present in the endometrium and placentomes of pregnant cows. Further analyses using rRNA gene sequencing techniques will aim to confirm the presence of bacteria in the bovine pregnant uterus and to investigate which species of bacteria are present in the uterus during pregnancy.