We recently developed a multilocus sequence typing (MLST) scheme to differentiate S. uberis isolates and facilitate an understanding of the population biology of this pathogen. The scheme was initially used to study a collection of 160 bovine milk isolates from the United Kingdom and showed that the majority of isolates were from one clonal complex (designated the ST-5 complex). Here we describe the MLST analysis of a collection of New Zealand isolates. These were obtained from diverse sources, including bovine milk, other bovine anatomical sites, and environmental sources. The complete allelic profiles of 253 isolates were determined. The collection was highly diverse and included 131 different sequence types (STs). The New Zealand and United Kingdom populations were distinct, since none of the 131 STs were represented within the previously studied collection of 160 United Kingdom S. uberis isolates. However, seven of the STs were members of the ST-5 clonal complex, the major complex within the United Kingdom collection. Two new clonal complexes were identified: ST-143 and ST-86. All three major complexes were isolated from milk, other bovine sites, and the environment. Carriage of the hasA gene, which is necessary for capsule formation, correlated with clonal complex and isolation from clinical cases of mastitis.

Microbiological and molecular tools were used to monitor Streptococcus uberis populations on farm tracks and paddocks on a dairy farm during different seasons of a year to identify and profile potential environmental niches of Strep. uberis in a pasture-based dairying system. Farm tracks of high or low cow traffic were sampled every 2 wk for an entire year and Strep. uberis numbers were enumerated from a selective medium. During each season of the year, paddocks were sampled for the presence of Strep. uberis before and after grazing by dairy cows. Farm tracks of high cow traffic generally had greater concentrations of Strep. uberis isolated compared with tracks with less cow traffic, but there was also significant variation in the concentrations of Strep. uberis contamination among seasons, being highest in winter and lowest in summer. The bacterium was detected in paddocks only after cow grazing had occurred, but the bacteria could still be detected in soil for up to 2 wk following grazing in winter. Multilocus sequence typing showed great heterogeneity, with some commonality between farm track and milk isolates, which may help explain cow-to-environment or environment-to-cow transmission of the bacterium in the dairy setting.

The objective of this study was to complete a positive-control, natural exposure, noninferiority design field study to test the efficacy of a novel glycolic acid-based postmilking teat disinfectant as compared with a previously proven iodine-based postmilking teat disinfectant (positive control). The primary outcome of interest was the effect of treatment on incidence of new intramammary infections. Secondary outcomes included the effect of treatment on prevalence of infection, somatic cell count, and teat condition. After blocking by parity, approximately 300 early- to mid-lactation cows on a large Wisconsin dairy farm were randomly assigned to 1 of 2 groups. For a 12-wk period between May and August 2014, the 2 groups were dipped after each milking with either the experimental (EX) or positive control (PC) product. Individual quarters were sampled to establish bacteriological infection status at the beginning of the study, and every 2 wk thereafter, by use of a 2-stage process evaluating somatic cell count (SCC), and then culturing milk samples only when SCC exceeded a parity-specific threshold. Teat condition scoring was completed at the beginning of the study and on wk 4, 8, and 12. Mixed logistic regression was used to evaluate the effect of treatment on dichotomous outcome measures including the odds of acquiring a new infection during a given 2-wk sampling interval (incidence), the odds for presence of infection at sampling (prevalence), and odds for a normal teat skin condition score. Mixed linear regression was used to evaluate the effect of treatment on somatic cell count. For the noninferiority analysis, the upper bound of the 95% confidence interval for the difference in new infection rate between the 2 treatments (EX - PC), had to be to the left of the critical value d (0.035) to conclude that EX was noninferior relative to PC with respect to risk for new infections. Results showed that the incidence of new infections was not different for quarters dipped with EX (3.2%) as compared with PC (4.2%). Similarly, the prevalence of infection tended to be lower for quarters dipped with EX (3.92%) as compared with PC (5.03%). No overall difference was found between treatments when evaluating somatic cell count measures and teat condition scores. Because the upper bound of the 95% confidence interval of the new IMI rate difference was smaller than the predefined noninferiority limit, it was concluded that the experimental product was not inferior compared with the positive control. As such, the glycolic acid-based teat disinfectant evaluated in this study can be considered an effective postmilking teat disinfectant, as well as safe, in so far as the product was not irritating to teat skin and did not negatively affect skin condition measures, as compared with the positive control group.

The objective of the study was to evaluate premilking teat disinfectant efficacy with the use of a novel diagnostic protocol against a positive control. The evaluation of efficacy was based on establishing noninferiority of a new premilking teat disinfectant compared with an existing premilking teat disinfectant. Approximately 200 cows were randomly allocated to 1 of 2 groups. Study personnel were blinded as to cow allocation and identification of the premilking disinfectants. Quarters were disinfected using a dip cup before milking with either the control or experimental product. The disinfectant was left on the teat for approximately 30s before being wiped off in preparation for milking. Quarter-level bacteriological infection status was established at the beginning of the study and quarter milk samples were collected biweekly thereafter. Bacteriological analyses were performed only when somatic cell counts in milk samples crossed a parity-specific threshold. Poisson regression models were used to analyze data. The difference in the rate of new intramammary infections was small and the upper bound of the 95% confidence interval of the rate difference was smaller than a predefined noninferiority limit. Based on the observed difference in new intramammary infections rate between the experimental and control product, it was concluded that the experimental disinfectant was not inferior compared with the positive control. The protocol used in this study proposes a valid and economically attractive methodology to evaluate the efficacy of teat disinfectants relative to a positive control.

The bacterial spirochetes, Treponema spp., are thought to be a major contributor to the etiology of bovine digital dermatitis (DD), a skin disease with worldwide economic impact. Hoofbath strategies are commonly used in an attempt to control and prevent the development of DD and continuing research has been done to develop an optimal hoofbath strategy for this purpose. The aim of this study was to develop a protocol that can be used as part of the screening process for candidate hoofbath disinfectants. This protocol allows an accurate determination of the in vitro minimum inhibitory concentration and minimum bactericidal concentration of a series of disinfectants for Treponema microorganisms. Assays were performed in triplicate for each of the disinfectants at 30-s and 10-min exposure times and exposed to 10 and 20% manure (vol/vol). The results of this study can be used to categorize disinfectants based on the effect of exposure and manure concentration regarding their ability to inhibit Treponema growth. This information can then aid in optimizing strategies for hoofbath-based control of DD development and spread.

A positive-control, natural exposure noninferiority field study was conducted to test the efficacy of a novel glycolic acid-based postmilking barrier teat disinfectant compared with a commercial iodine-based postmilking barrier teat disinfectant (positive control). Cows from 2 pens from a California Central Valley dairy farm were dipped after milking either with the positive-control product (PC) or the experimental product (EX) over 12 wk. New intramammary infections (NIMI) were determined by biweekly sampling of all quarters of study cows and classified as a NIMI based on somatic cell count and milk bacteriological culture results. The mean quarter-level incidence risks during a 2 wk study period were 3.50% (EX) and 4.28% (PC). The majority of NIMI were caused by coagulase-negative staphylococci, followed by non-agalactiae streptococci. The study results indicated that EX was noninferior to PC, with a 17% relative efficacy (improvement) in reducing NIMI compared with the PC group. Also, quarter somatic cell count was not affected by the postmilking teat disinfectant used. Finally, the EX product was safe in terms of teat conditioning: teat condition scores were not different between study groups. The study concluded that the glycolic acid-based experimental post-dip barrier was noninferior to the control, and could be considered a safe and effective postmilking teat disinfectant.

The objectives of this research were to test the potential of unsupervised (USNN) and supervised neural network (SNN) models for detecting major and minor mastitis pathogens based on changes in milk parameters. A data set of 4,852 quarter milk samples with records for milk parameters and bacteriological status was used to train and validate the models by classifying milk samples into 3 different bacteriological states: not infected, intramammary infection (IMI) by minor pathogens, and IMI by major pathogens. Sensitivity of the USNN model was 97% for detecting noninfected quarters, 89% for minor pathogen IMI, and 80% for major pathogen IMI. Specificities of USNN models were close to 99% for all bacteriological states. The sensitivity of SNN models was affected by the ratio of infected to noninfected cases in the data set. As the ratio of infected to healthy cases increased from 1:1 to 1:10, detection accuracy for noninfected quarters increased from 82 to 98% but that for minor pathogen IMI decreased from 86 to 44%. The sensitivity for major pathogen IMI was 20% when the ratio was 1:1, but ranged from 20 to 40% when different ratios were tested. The SNN models indicated that somatic cell score and electrical resistance index had the most discriminating power. It was concluded that both USNN and SNN models were able to effectively differentiate between noninfected quarters and those infected by minor mastitis pathogens, and that the USNN model had a better agreement with results obtained from conventional microbiological methods. These types of models can be used in in-line milking systems to detect the infection status of a quarter and provide the farmer with diagnosing options for managing mastitis.

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