A New Selective Medium for Detection of Klebsiella from Dairy Environments

Abstract

Klebsiella causes severe mastitis in dairy cattle and is a great concern for dairy farm management. The media previously used to detect Klebsiella, MacConkey-Inositol-Carbenicillin agar (MCIC) and Simons-Citrate agar supplemented with 1% inositol (SCAI), were not selective enough for samples from the dairy environment such as cow feces or cow bedding materials. We developed a new selective medium for Klebsiella. BIND (Brilliant green containing Inositol-Nitrate-Deoxycholate agar) medium contains myo-inositol and sodium nitrate as a sole carbon and nitrogen source, respectively, and is supplemented with brilliant green and deoxycholate. All Klebsiella strains isolated from the dairy environment grew well on the BIND plates. The growth of non-Klebsiella contaminants, which grew well on MCIC or SCAI, was effectively suppressed on BIND. All isolates that grew on BIND were identified as Klebsiella both by API testing and by 16S rDNA sequence alignment. We concluded that BIND was selective enough for the detection of Klebsiella species from dairy samples and therefore, helpful for monitoring Klebsiella populations in the dairy environment and for preventing the mastitis caused by Klebsiella.