Influence of teat canal keratin on susceptibility to intramammary infection was investigated in lactating Jersey cows. In each of two replicate trials, keratin was removed from the left teats of 20 cows immediately before milking. Immediately after milking, all teats were exposed to bacterial challenge by immersion in a suspension of Streptococcus agalactiae (5 x 10(7) cfu/ml). Bacterial challenge was repeated after the next four milkings. Foremilk samples were obtained for 8 d after keratin removal to determine infection status. A mammary quarter was classified as infected based solely upon the bacteriological criteria outlined by the National Mastitis Council. The rate of infection in quarters from which keratin was removed was greater than that in control quarters. Infection rates were 26.3% for keratin-removed quarters and 8.3% for control quarters in trial 1 and 13.5 and 0%, respectively, in trial 2. When more stringent criteria (recovery of greater than 100 cfu of S. agalactiae/ml in three or more successive milk samples and a SCC of greater than 10(6)) were used to identify a subset of infections that were clearly intramammary, infection rates were 9.3% for keratin-removed quarters and 1.4% for control quarters. Thus, partial removal of keratin from the teat canal compromised the ability of the teat to prevent passage of bacterial pathogens from the external environment into the mammary gland.

Twenty-four Holstein cows, producing at least 21 kg of milk/d, were used in two replicate experiments to determine the effect of presence or absence of pulsation on loss of teat canal keratin during machine milking. Left quarters were milked without pulsation and right quarters were milked with pulsation. On d 0 and 10, keratin was collected from one left and from one right teat canal of each cow prior to milking and from the remaining two teat canals after milking. Milk was collected for assessment of SCC and bacteriological status on d 0 and approximately every 3 d until d 18. Quantity of keratin recovered before milking on d 10 did not differ between teats milked with or without pulsation, but loss of keratin because of milking was greater from teats milked with pulsation. By d 7, 30% (12 of 43) of quarters milked without pulsation had become infected, but no (0 of 47) quarters milked with pulsation were infected. By d 14 to 16, new infections had increased to 68% (28 of 41) of quarters milked without pulsation and 2% (1 of 43) in quarters milked with pulsation; mean SCC in pulsationless quarters increased sevenfold relative to pulsation quarters. Protein and water content of keratin did not differ because of treatment, and changes in lipid composition were minor. Histological analysis of the teats of 4 cows indicated that the mean diameter of the teat canal, within 2 h after milking, was greater without pulsation than with pulsation (680 vs. 483 microns).

Methods for collecting keratin from the teat canal were examined to select a procedure to obtain representative samples for lipid analysis. Data obtained by solvent extraction of excised teats were compared with those obtained by scraping keratin from dissected teats of lactating and dry cows. Solvent extraction with petroleum ether or 2:1 chloroform-methanol yielded similar dry weights of material. However, both solvents removed large amounts of material other than keratin from the teat canal. The lipid class and fatty acid profiles of the material extracted by solvent flushing were not similar to profiles obtained by scraping keratin from the teat canal. A metal tapestry needle was suitable for collection of keratin from the teat canal of living cows. About 78% of the keratin present in the teat was collected with the needle. Lipid composition of keratin collected with the needle was the same as in keratin scraped from excised teats. The tapestry needle was suitable as a tool for collecting repeatable, representative samples of keratin for analysis from single teat canals of living cows.

This study was designed to determine the regeneration rate of teat canal keratin in two groups of 10 lactating Holstein cows. The weight of keratin obtained upon successive collections of keratin was determined. Intervals between successive collections were varied between 8 and 72 h. Following initial collection, the quantity of keratin removed was regenerated within 1 to 2.5 d. The rate of keratin regeneration per teat was approximately 1.5 mg of wet weight or .6 mg of dry weight per 24 h. The concentration (microgram lipid/mg keratin, wet weight) of lipid in keratin samples collected during regeneration was almost twice that present initially or after regeneration was complete. With the exception of free cholesterol, composition (%) of lipid components of keratin did not differ between samples collected initially, those collected early in the process of regeneration, and those collected after the mass of keratin removed had been replaced. Although data suggest that keratin matures during the process of regeneration, maturation appears complete by the time mass of keratin is fully regenerated. Additionally, the quantity of keratin recovered at initial sampling was inversely correlated with milk production. We hypothesize that as milk production increases, more keratin is lost during milking.