Objective—To evaluate enterotoxin production, enterotoxin gene distribution, and genetic diversity of Staphylococcus aureus in milk obtained from cows with subclinical mastitis. Sample—Milk samples obtained from 350 cows (1,354 mammary glands) on 11 Wisconsin dairy farms. Procedures—Of 252 S aureus isolates obtained from 146 cows, 83 isolates (from 66 cows with subclinical mastitis) were compared genotypically by use of pulsed-field gel electrophoresis and via PCR identification of toxic shock syndrome toxin 1 (TSST-1) and classical S aureus enterotoxin genes (sea, seb, sec, sed, and see). Results—Among the 83 S aureus isolates, ≥ 1 enterotoxin genes were identified in 8 (9.6%). Enterotoxin gene distribution was as follows: TSST-1, 7 isolates (8.4%); sec, 5 isolates (6.0%); and sed, 2 isolates (2.4%). Enterotoxin genes sea, seb, and see were not identified. Twelve pulsotypes and 5 subtypes were identified among the 83 isolates; 5 of the 12 pulsotypes were represented by only 1 isolate. In cows of 1 herd, only a single S aureus pulsotype was detected; in cows on most other farms, a variety of pulsotypes were identified. One pulsotype was recovered from 4 farms (n = 23 cows) and another from 5 other farms (16). Isolates with an enterotoxin gene were represented by 6 pulsotypes. Conclusions and Clinical Relevance—S aureus classical enterotoxins and TSST-1 were rarely recovered from milk samples obtained from cows with subclinical mastitis in Wisconsin. Diverse pulsotypes of S aureus were detected within and among farms, indicating that different strains of S aureus cause subclinical mastitis in dairy cows.

Objective—To compare results for 3 commercially available microbiological media plates with those for standard bacteriologic testing of bovine milk. Sample—Milk samples from postpartum cows and cows with a high somatic cell count (SCC) or clinical mastitis (CM). Procedures—Sample-ready Staphylococcus culture medium (SRSC) plates were used to detect Staphylococcus aureus in milk samples obtained from postpartum cows and cows with a high SCC or CM. Rapid coliform count (RCC) plates were used to detect coliforms in milk samples obtained from cows with CM. Aerobic count (AC) plates were used to detect streptococci in CM samples. Fresh mastitic milk samples were frozen and then thawed to evaluate the effects of freezing for the SRSC and RCC plates. The effects of dilution (1:10) of samples were determined. Agreement of results between the commercially available plates and standard bacteriologic testing was evaluated. Results—The ability of SRSC plates to detect S aureus in milk samples was highest with diluted samples from postpartum cows and cows with a high SCC or CM. Sensitivity of the RCC plate for detection of coliforms was highest with diluted mastitic milk samples. The AC plates had a poor positive predictive value for detection of streptococci in mastitic milk samples. Freezing increased S aureus detection. Conclusions and Clinical Relevance—Overall, the SRSC and RCC plates were accurate, were easy to use, and yielded results comparable to those of standard bacteriologic testing for the detection of S aureus and coliforms in bovine milk.