Rapid determination of pathogens in mastitic milk of dairy cows using Gram staining

This study aimed to determine whether causative pathogens in mastitic milk can be determined by Gram staining after the centrifugation of milk. Gram staining was performed using unconcentrated and concentrated milk cells. Using this method, we found that the background of microscopic image of unconcentrated milk cells was complex and bacteria were difficult to detect. In contrast, the background of the smears in the concentrated milk cells was translucent, and bacterial and somatic cells were clearly visible. The sensitivity and specificity of the Gram staining of concentrated milk cells were 84.4% and 86.0% and 50.0% and 94.5% for the detection of gram-positive and gram-negative bacteria, respectively. The presented method provides a simple and inexpensive means of determining mastitis-causing pathogens.