Molecular epidemiology of nonsevere clinical mastitis caused by Klebsiella pneumoniae occurring in cows on 2 Wisconsin dairy farms

Abstract

The objective of this study was to describe diversity of Klebsiella pneumoniae isolated from milk collected at detection of nonsevere (abnormal milk or abnormal udder) clinical mastitis (CM) and during a follow-up period. Cases were detected in cows enrolled in a randomized clinical trial (n = 168) conducted using 2 related Wisconsin dairy farms. Cases were randomly assigned to receive 2 d (n = 18) or 8 d (n = 18) of intramammary infusions with an approved product containing ceftiofur hydrochloride or assigned to a negative control group (n = 17). Milk samples were collected from affected quarters at detection and during a 28-d follow-up period. Sufficient DNA was recovered from 53 of 54 Kleb. pneumoniae cultured from quarter milk samples collected at detection of the incident case. Additional Kleb. pneumoniae were recovered from milk samples collected from the same quarters at 14, 21, and 28 d after case detection (n = 35), at detection of recurrent cases in the same quarter (n = 14), and from new cases of CM (n = 3) occurring in enrolled quarters. All Kleb. pneumoniae were used for molecular typing by pulsed-field gel electrophoresis and 90% similarity was used to define homology. Of Kleb. pneumoniae recovered from incident cases, unique strains (n = 41) were identified in milk samples collected from cows on farm A (n = 19) and farm B (n = 22), whereas 12 clonal strains were identified with 8 found only in milk collected from farm A and 4 found in milk samples collected from cows on both farms. Heterogeneous strains of Kleb. pneumoniae genotypes were isolated from incident cases of CM. However, when intramammary infection persisted or when recurrence of CM occurred, clonal strains were isolated at 14, 21, or 28 d. Similar strains of Kleb. pneumoniae genotypes caused persistent CM. In conclusion, initial cases of CM were caused by a wide genetic diversity of Kleb. pneumoniae, but when IMI persisted, the same strain often persisted within the mammary gland up to 28 d.