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Yellowish discharge after application of intravaginal progesterone releasing inserts is frequently observed in cows. The objective of this study was to compare the bacteriological contamination of the vagina and uterus before and after a treatment with a progesterone insert in heifers. Forty-two Holstein heifers received a progesterone releasing insert [Eazi-Breed controlled internal drug release (CIDR) insert; Pfizer Animal Health, Berlin, Germany] for 7d. The protruding tail had been removed from half of the inserts (no tail group: n=21; tail group: n=21). Nine heifers from the tail group lost the insert within the 7-d treatment interval and were excluded. Heifers identified in estrus were artificially inseminated on d 9 or 10. Vaginal discharge was scored on a 4-point scale [vaginal discharge score (VDS) 0 to 3] and vaginal swabs were taken for bacteriological examination on d 0 and 7 and the day of artificial insemination (AI). Furthermore, cytological and bacteriological samples were obtained from the uterus on d 7 and the day of AI. On d 0, coliforms and Streptococcus spp. were found in vaginal swabs of 21 heifers (64%). On d 7, all heifers showed purulent vaginal discharge (VDS 2 to 3). The VDS was higher in the tail group compared with the no tail group. Arcanobacterium pyogenes, coliforms, and Streptococcus spp. were isolated from the vaginal swabs in 32 of 33 (96%) heifers on d 7. On the day of AI, VDS had improved to 0 or 1 in 96% of the heifers. However, A. pyogenes, coliforms, and Streptococcus spp. were still isolated in 17 of 33 (53%) heifers from the vagina and in 32 of 33 (96%) heifers from the endometrium. Endometrial cytology revealed polymorphonuclear neutrophils (PMN) in 11 heifers (6 to 32% PMN). Five samples exceeded the threshold of 5% PMN, and 2 samples exceeded the 10% PMN threshold, indicative of subclinical endometritis. In conclusion, pyogenic bacteria were found in the vagina and uterus on d 7 and the day of AI after intravaginal progesterone treatment. The severity of the discharge was affected by the protruding tail of the insert.