Compliance with topical timolol treatment.

To determine if compliance with timolol treatment was better than compliance with pilocarpine treatment (as reported previously), we measured compliance with timolol treatment in a sample of 110 patients using an unobtrusive eyedrop medication monitor, which recorded electronically the date and time of each drug administration over a four- to six-week period. The patients administered a mean +/- S.D. of 82.7% +/- 19.0% of the prescribed timolol doses (range, 20% to 100%). Forty-five patients were treated concurrently with timolol and pilocarpine. These patients administered a mean +/- S.D. of 84.3% +/- 14.0% of the prescribed timolol doses and 77.7% +/- 18.7% of the prescribed pilocarpine doses (P = .012, van der Waerden test). Our results suggest that while compliance is influenced by the drug regimen, defaulting is not eliminated by prescribing a more convenient medication with fewer side effects.

Use of predictive modeling to evaluate the manipulation of milking frequency, temperature, and oxygen tension on growth of Escherichia coli in an artificial intramammary environment.

A method was developed to evaluate frequent milking as a means of controlling intramammary infection. An artificial intramammary environment was used to determine growth responses of Escherichia coli (P4) to natural changes in the mammary gland resulting from bacterial invasion. Physical conditions manipulated in this model were growth medium, temperature, and oxygen tension. Mathematical modeling was then incorporated to generate predictions concerning growth dynamics of the organism when milking frequency was changed. To test accuracy of the model, initial predictions were derived from bacterial growth data in which E coli was incubated in tryptose soy broth for 12 hours at 37 C and PO2 equal to 23.3 mm of Hg. These predictions matched closely with experimental data in which 12-, 4-, and 2-hour milking intervals were simulated in the artificial intramammary environment. The mathematical model was then used to characterize growth rate data from in vitro experiments in ultra-high temperature-treated milk and in vivo experimental infection data generated with E coli (P4). Predictions generated from this model suggested that increasing milking frequency to 4 or 6 times daily controls growth of E coli for a prolonged period and that 12 times daily milking may lead to elimination of the bacterium.

Effects of temperature and oxygen tension on growth of Escherichia coli in milk.

An in vitro system was developed to mimic conditions within normal and mastitic mammary glands. The system consisted of a modified batch fermenter that allowed for manipulation of temperature, oxygen tension, and pH. Experiments in tryptose-soy broth and UHT-treated milk were conducted to evaluate growth characteristics of Escherichia coli P4 as physical conditions were manipulated. The effect of bacterial growth on oxygen tension and pH were also evaluated. Growth of E. coli was inhibited as temperature was increased from 37 to 41 degrees C and as oxygen tension was decreased from microaerophilic to anaerobic levels. At bacterial populations > 6 log10 cfu/ml, microaerophilic cultures became anaerobic. The pH followed a similar trend; however, after a significant decrease in pH, mean bacterial populations were 7.1 log10 cfu/ml in tryptosesoy broth and 8.2 log10 cfu/ml in UHT-treated milk. This dynamic model demonstrated potential use in evaluation of growth characteristics of mammary gland pathogens in the lactating mammary gland.

Effect of oxygen tension on killing of Escherichia coli by bovine polymorphonuclear neutrophil leucocytes in vitro.

A batch fermenter modified to simulate the physical conditions of an inflamed v. uninflamed mammary gland was utilized to evaluate the effect of oxygen tension on killing of Escherichia coli P4 by bovine polymorphonuclear neutrophil leucocytes. Leucocytosis was simulated in vitro 4 h after inoculation with Escherichia coli by adding bovine neutrophils isolated from peripheral blood to the culture medium. Experiments were conducted at 39 degrees C in UHT treated milk. At micro-aerophilic oxygen tension (oxygen partial pressure, 3.11 kPa), bacterial numbers declined during the first hour following addition of the neutrophils. Oxygen tension declined rapidly following PMN addition. Once oxygen was depleted, neutrophil activity was presumably diminished and Esch. coli numbers began to increase. Under anaerobic conditions (oxygen partial pressure, 0.17 kPa), no reduction in population was observed. Photomicrographs taken at the time of neutrophil addition and at subsequent time intervals demonstrated a specific association between neutrophils and the pathogen. Subsequent lysis of neutrophils associated with Esch. coli growth was seen coincident with oxygen depletion.

Effect of individual caseins on plasminogen activation by bovine urokinase-type and tissue-type plasminogen activators.

The effect was examined of individual caseins on the rate of plasminogen activation by bovine urokinase-type and tissue-type plasminogen activators. All individual caseins (alpha-CN, beta-CN, and kappa-CN) enhanced the activity of both types of plasminogen activators. Optimal concentrations for alpha-CN and beta-CN were 5 and 25 micrograms/ml, respectively. The enhancement of enzymatic activity declined when concentrations of alpha-CN and beta-CN were higher. In contrast, increasing concentrations of kappa-CN from 0 to 200 micrograms/ml resulted in corresponding increases in activity of both types of plasminogen activators. On a weight basis, alpha-CN was the most effective enhancer of plasminogen activator activity. Indirect evidence obtained with experiments utilizing alpha-CN immobilized on agarose suggested that the effect is related to extensive binding of plasminogen and both types of plasminogen activators to casein.

Hormonal and extracellular matrix regulation of plasminogen activator in a bovine mammary epithelial cell line.

The effects of lactogenic hormones on urokinase plasminogen activator (u-PA) produced by bovine mammary epithelial cells (MAC-T) were examined. High levels of u-PA activity were detected in growth arrested cells cultured on plastic. This suggests that high levels of PA activity alone are not sufficient to induce proliferation of bovine mammary epithelial cells. Cells were cultured on various extracellular matrices: plastic, fibronectin, collagen, matrigel, and laminin. Basal levels of u-PA activity in media from MAC-T cells cultured on matrigel were 1.6-, 2-, 2- and 3.5-fold higher than that of cells cultured on plastic, fibronectin, collagen, and laminin, respectively. Insulin increased (P<0.01) mammary epithelial u-PA activity on a per cell basis, an effect observed irrespectively of the type of extracellular matrix onto which cells were cultured. Not unexpectedly, insulin-like growth factor (IGF)-I, Des(1-3) IGF-I and IGF-II increased mammary epithelial u-PA activity on a per cell basis and u-PA mRNA levels, thus, mimicking the effect of insulin. Dexamethasone suppressed (P<0.01) u-PA activity but was unable to suppress the insulin-induced increase in u-PA activity of cells cultured on various extracellular matrices. These data indicate that u-PA activity is modulated by both lactogenic hormones and the type of extracellular matrix.

The influence of intensively managed rotational grazing, traditional continuous grazing, and confinement housing on bulk tank milk quality and udder health.

Monthly bulk tank milk samples and veterinary records were analyzed for 1 yr on 15 Vermont dairy farms. Data were evaluated using ANOVA to compare effects of grazing management systems on milk quality and udder health. Systems evaluated were intensively managed rotational grazing, traditional continuous grazing, and confinement housing. Bulk tank samples were evaluated for standard plate count, bacterial type counts on tryptose-blood-esculin agar, and SCC. Veterinary records were evaluated for incidence of clinical mastitis, udder edema, and teat injuries. Within- and between-treatment group analyses were conducted by season, herd size, and udder sanitation systems. Mean standard plate counts were lower in rotationally grazed herds than counts of confined herds during the grazing season. Similarly, rotationally grazed herds with fewer than 60 cows had lower standard plate counts than confined herds of similar size. Mean bulk tank counts of streptococci other than Streptococcus agalactiae during the grazing season differed among treatments. The lowest counts occurred in rotationally grazed herds. Among herd using predip products recognized as efficacious, fewer streptococci other than S. agalactiae were isolated from bulk tank milk of rotationally grazed herds than confined herds. Rotationally grazed herds using postdips recognized as efficacious had lower SCC than those using unrecognized postdips. No udder health differences were observed among grazing treatments.

Evaluation of a 1% iodophor postmilking teat sanitizer.

A natural exposure field trial a with positive control was conducted to evaluate bacteriological efficacy and teat conditioning qualities of an experimental postmilking teat dip. An experimental 1% iodine postmilking teat sanitizer with a 10% emollient system was compared with a 1% iodine plus 10% glycerin teat sanitizer. Efficacy of the two sanitizers was equivalent for all new IMI, major pathogens, and environmental pathogens. The products were not equivalent for efficacy against coliforms and coagulase-negative staphylococci. Fewer coliform IMI were diagnosed in the control group than in the treatment group. Differences were determined for efficacy against coagulase-negative staphylococci in favor of the treatment product. The products were equivalent for all clinical mastitis, including previously existing IMI that became clinical. The products were not equivalent for all or new clinical IMI with major pathogens, all environmental pathogens, or coliforms. Fewer infections were diagnosed in the control group than in the treatment group. Teat end and teat skin conditions improved with the use of the triple emollient, postmilking teat sanitizer under the winter conditions experienced during this field trial.

Winter evaluation of a postmilking powdered teat dip.

A powdered teat dip designed for winter usage was evaluated for bacteriological efficacy and teat conditioning qualities. A positive control, natural exposure field trial was conducted for 3 mo on 509 lactating cows. Two sets of cows, primiparous and multiparous, were used. The trial compared efficacy of a powdered teat dip with a teat dip of 1% iodine plus 10% glycerin. Bacteriological efficacy among primiparous cows was equivalent for all major mastitis pathogens, environmental pathogens, and streptococci other than Streptococcus agalactiae. Efficacy was not equivalent against coagulase-negative staphylococci and all mastitis pathogens. Results suggested that the positive control product was more efficacious. Among multiparous cows, efficacy was equivalent against environmental mastitis pathogens and bacteriologically negative, clinical mastitis. The products were not equivalent against Staphylococcus aureus, coagulase-negative staphylococci, or all major mastitis pathogens, once again suggesting that the positive control product was more efficacious. Data indicated that germicidal activity of the powdered dip was not sufficient to reduce the incidence of new IMI caused by contagious or minor pathogens normally associated with teat skin. Application of a powdered postmilking teat dip during 3 winter mo in Idaho resulted in improved teat end condition among primiparous and multiparous dairy cows. Teat skin condition improved among primiparous but not among multiparous cows.