The objectives of the present study were to investigate the bacteria accompanying hydrosalpinx of the buffalo cow and investigate the correlation between bacterial infection of the uterus and hydrosalpinx. Buffalo cows' reproductive tracts were collected from Mosul abattoir. A total 385 uterine samples were examined of which 25 were having hydrosalpinx. Swabs for bacteriology, fluid for cytology and biopsies for histopathology were collected from the hydrosalpinx and the uterus from each samples included in this study. Results of this study indicated high prevalence of hydrosalpinx (6.5%) including unilateral (n = 19; 76%) and bilateral (n = 6; 24%) hydrosalpinx. Although 16 samples (64%) of the hydrosalpinx samples had no bacterial growth, the most prevalent bacteria recovered from hydrosalpinx were Corynebacterium hemolyticum and Actinomyces bovis, 42.8% and 28.6%, respectively. The most prevalent bacteria in the uterus were Archanobacterium pyogenes (18.5%), Staphylococcus aureus (14.8%), and Listeria monocytogenes (11.0%). Higher rates of leukocytes infiltration (p < 0.01) were observed in the uterine discharge than hydrosalpinx. A significant (p < 0.01) increase in lymphocytes was found in uterine discharge. Microscopic examination of the hydrosalpinx showed mucosal atrophy and dilatation of oviductal lumen without any signs of inflammation. It could be concluded that there is no correlation between bacteria isolated from uterus and hydrosalpinx. No association was found between bacteriological cultures and hydrosalpinx. Inflammation of the uterine tissue could be extended to utero-tubal junction producing local inflammation resulting in fibrosis and tubal obstruction. The obstruction in the lumen of the oviducts resulted in accumulation of fluid.