Pegbovigrastim treatment resulted in an economic benefit in a large randomized clinical trial in grazing dairy cows.

This randomized controlled trial on 4 commercial grazing dairy farms investigated whether pegbovigrastim (PEG) treatment affected partial net return as calculated from milk revenues and costs for feed, medical treatments [clinical mastitis, uterine disease, and other diseases (i.e., any medical treatment that was not intended for clinical mastitis or uterine disease)], inseminations, and culling during a full lactation in grazing dairy cows. We also explored the effect of potential interactions of PEG treatment with parity, prepartum body condition score, and prepartum nonesterified fatty acids concentration on partial net return, milk revenues, and the costs mentioned above. Holstein cows were randomly assigned to 1 of the 2 following trial arms: a first PEG dose 9.4 ± 0.3 (mean ± standard error) days before the calving date and a second dose within 24 hours after calving (PEG: primiparous = 342; multiparous = 697) compared with untreated controls (control: primiparous = 391; multiparous = 723). The effect of PEG treatment on the outcomes of interest expressed per year was tested using general linear mixed models. Results are presented as least squares means ± standard error. Overall, PEG treatment increased the partial net return, resulting in an economic benefit per cow per year of $210 ± 100. The cost of treatment of clinical mastitis was lower for PEG treated cows compared with control cows ($9 ± 3). The largest nonsignificant difference was seen for the cost of culling; additionally, PEG treatment numerically reduced the cost of culling by $145 ± 77.

Smallholder milk-quality awareness in Indonesian dairy farms.

In most low- and middle-income countries, milk is produced by smallholders, thereby contributing to the livelihood of their households. With the increasing importance of milk production in these countries, it is essential that milk quality is of a high level to ensure a safe product for consumers. It is, however, unclear whether smallholder dairy farmers are aware of the quality of their milk. The aim of this cross-sectional study was to gain insight on Indonesian smallholder dairy farmer awareness of milk quality parameters and to identify factors associated with the total plate count (TPC) and somatic cell count (SCC). A stratified sampling method was used to select smallholder farms in 4 districts in West Java, Indonesia, that were interviewed between August and September 2017. Factors putatively associated with awareness of TPC were investigated with multinomial regression models, whereas a Firth-type logistic regression was applied to identify factors associated with SCC awareness. Of the total 600 farmers surveyed, 264 (44%), 109 (18%), 170 (28%), 111 (19%), and 23 (4%) farmers were aware of TPC, total solid, fat content, milk density, and SCC, respectively, but did not know its value. Those that were conceptually aware of these quality parameters were generally unaware of their value. Furthermore, this study revealed that the following variables were significantly associated with dairy farmers' awareness of TPC: cooperative to which the farmer belonged, distance to neighboring dairy farmer, technology adoption index, TPC as the most important quality factor for the buyer, milk production information from cooperatives, and cow health information from veterinarians. Similarly, cooperative, dairy business experience, and milk quality test adoption were significantly associated with dairy farmers' awareness of SCC. Cooperative was the only variable that was significant in both final statistical models. This indicates that cooperatives play an important role in increasing farmer awareness of milk quality parameters in these smallholder dairies. This may be valid for other regions in the world also where milk production is dominated by smallholder dairy farmers.

Effect of pegbovigrastim on fertility and culling in grazing dairy cows and its association with prepartum nonesterified fatty acids.

This randomized controlled trial on 4 commercial grazing dairy farms investigated whether treatment with pegbovigrastim (PEG) affected fertility and culling as measured during the full lactation. We also explored the effect of potential interactions of PEG treatment with parity, prepartum body condition score, prepartum nonesterified fatty acid concentration (pre-NEFA), and early-lactation clinical disease on these outcomes. Holstein cows were randomly assigned to 1 of 2 trial arms: a first PEG dose approximately 7 d before the expected calving date and a second dose within 24 h after calving (PEG: primiparous = 342; multiparous = 697) compared with untreated controls (control: primiparous = 391; multiparous = 723). Cox's proportional hazards regression models were used to analyze rate of first insemination, rate of pregnancy [within 150 and 305 d in milk (DIM)], and hazard of culling. Additional analyses were performed on data that were stratified by parity group and pre-NEFA class (low ≤0.3; high >0.3 mM). In high pre-NEFA cows, PEG treatment increased the rate of first insemination [hazard ratio (HR) = 1.15]. Early-lactation clinical mastitis (CM) and uterine disease (UD: retained placenta, metritis, or both) were associated with a reduced rate of pregnancy within 150 DIM (HR = 0.49 and 0.78, respectively). Pegbovigrastim treatment in high pre-NEFA cows with CM and UD increased the rate of pregnancy within 150 DIM (HR = 1.75 and 1.46, respectively). In high pre-NEFA cows, PEG treatment resulted in a lower hazard of culling (HR = 0.79). No treatment effect was detected in low pre-NEFA cows. This study shows that the effect of PEG treatment on fertility and culling interacts with pre-NEFA. In high pre-NEFA cows, PEG treatment increased the rate of first insemination, counteracted the negative association of early-lactation CM and UD with the rate of pregnancy, and decreased the hazard of culling.

Dynamics of somatic cell count patterns as a proxy for transmission of mastitis pathogens.

Management of udder health is particularly focused on preventing new infections. Data from the DeLaval Online Cell Counter (DeLaval, Tumba, Sweden) may be used in forecasting to improve decision support for improved udder health management. It provides online cell counts (OCC) as a proxy for somatic cell counts from every milking at the cow level. However, these values are typically too insensitive and nonspecific to indicate subclinical intramammary infection (IMI). Our aim was to describe and evaluate use of dynamic transmission models to forecast subclinical IMI episodes using milk cultures or changes in OCC patterns over time. The latter was expressed by an elevated mastitis risk variable. Data were obtained from the dairy herd of the Norwegian University of Life Sciences (Oslo, Norway). In total, 173 cows were sampled monthly for bacteriological milk culture during a 17-mo study period and 5,330 quarter milk samples were cultured. Mastitis pathogens identified were assigned to 1 of 2 groups, Pat 1 or Pat 2. Pathogens from which a high cell count would be expected during a subclinical IMI episode were assigned to the Pat 1 group. Pathogens not in the Pat 1 group were assigned to the Pat 2 group. Staphylococcus epidermidis, Staphylococcus aureus, and Streptococcus dysgalactiae were the most common Pat 1 pathogens. Corynebacterium bovis, Staphylococcus chromogenes, and Staphylococcus haemolyticus were the most common Pat 2 pathogens. The OCC were successfully recorded from 82,182 of 96,542 milkings. The current study included 324 subclinical IMI episodes. None of the mastitis pathogens demonstrated a basic reproduction number (R0) >1. Patterns of OCC change related to an episode of Pat 1 subclinical IMI at specificity levels of 80, 90, and 95% at sensitivity levels of 69, 59, and 48% respectively, demonstrated an R0 >1. An existing infection was significant for transmission for several Pat 2 pathogens, but only for Staphylococcus aureus and Staphylococcus epidermidis among Pat 1 pathogens. Dynamic transmission models showed that patterns of OCC change related to an episode of Pat 1 subclinical IMI were significantly related to the same pattern occurring in susceptible cows at specificity levels of 80, 90, and 99% at sensitivity levels of 69, 48, and 8%, respectively. We conclude that changes in herd prevalence of subclinical IMI can be predicted using dynamic transmission models based on patterns of OCC change. Choice of specificity level depends on management goals and tolerance for false-positive alerts.

The detection of intramammary infections using online somatic cell counts.

Timely and accurate identification of cows with intramammary infections is essential for optimal udder health management. Various sensor systems have been developed to provide udder health information that can be used as a decision support tool for the farmer. Among these sensors, the DeLaval Online Cell Counter (DeLaval, Tumba, Sweden) provides somatic cell counts from every milking at cow level. Our aim was to describe and evaluate diagnostic sensor properties of these online cell counts (OCC) for detecting an intramammary infection, defined as an episode of subclinical mastitis or a new case of clinical mastitis. The predictive abilities of a single OCC value, rolling averages of OCC values, and an elevated mastitis risk (EMR) variable were compared for their accuracy in identifying cows with episodes of subclinical mastitis or new cases of clinical mastitis. Detection of subclinical mastitis episodes by OCC was performed in 2 separate groups of different mastitis pathogens, Pat 1 and Pat 2, categorized by their known ability to increase somatic cell count. The data for this study were obtained in a field trial conducted in the dairy herd of the Norwegian University of Life Sciences. Altogether, 173 cows were sampled at least once during a 17-mo study period. The total number of quarter milk cultures was 5,330. The most common Pat 1 pathogens were Staphylococcus epidermidis, Staphylococcus aureus, and Streptococcus dysgalactiae. The most common Pat 2 pathogens were Corynebacterium bovis, Staphylococcus chromogenes, and Staphylococcus haemolyticus. The OCC were successfully recorded from 82,182 of 96,542 milkings during the study period. For episodes of subclinical mastitis the rolling 7-d average OCC and the EMR approach performed better than a single OCC value for detection of Pat 1 subclinical mastitis episodes. The EMR approach outperformed the OCC approaches for detection of Pat 2 subclinical mastitis episodes. For the 2 pathogen groups, the sensitivity of detection of subclinical mastitis episodes was 69% (Pat 1) and 31% (Pat 2), respectively, at a predefined specificity of 80% (EMR). All 3 approaches were equally good at detecting new cases of clinical mastitis, with an optimum sensitivity of 80% and specificity of 90% (single OCC value).

Transmission dynamics of intramammary infections caused by Corynebacterium species.

The development of reliable models for transmission of intramammary infections (IMI) is the subject of extensive research. Such models are useful to enhance the identification and understanding of factors that affect pathogen-specific IMI dynamics. Longitudinal transmission models are valuable for predicting infection outbreak risks, quantifying the effectiveness of response tactics, and performing response planning. In this work, we focused on modeling Corynebacterium spp. by using a compartmental model. Previous investigations have considered modeling the transmission dynamics of several bacterial pathogens, but not Corynebacterium spp. We established a Corynebacterium spp. Susceptible-Infectious-Susceptible (SIS) model. We simulated the model numerically by using parameters that we estimated by a generalized linear model approach, using month of study as the time variable. The data, from which the parameters of the model were estimated, were obtained in a field trial conducted in 2 US dairy herds. Altogether, 786 cows were sampled at least once during the 13-mo study period. The total number of quarter milk cultures and cases of IMI caused by Corynebacterium spp. were 11,744 and 556, respectively, in farm A; the corresponding figures for farm B were 11,804 and 179. Our modeling study included only transmission from persistent IMI caused by Corynebacterium spp. within the lactation pens. The rate of new infections was significantly related to preexisting IMI in both farms, underscoring the importance of preexisting Corynebacterium spp. IMI for the transmission of Corynebacterium spp. within lactation pens. The estimated basic reproduction numbers (R0) in the 2 farms were 1.18 and 0.98, respectively. The nonsignificant disparity in R0 was associated with significant differences in cure rates between farms.

Modeling of Mycobacterium avium subsp. paratuberculosis dynamics in a dairy herd: An individual based approach.

In the dairy industry, Johne's disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (MAP) is one of the major investigated diseases. To date, researchers have suggested some control strategies for JD, such as test-and-cull based herd management, isolated calf rearing management, and vaccinations. Due to the slow progressing nature of MAP, tests with low diagnostic test sensitivity and specificity, and economic limitations, implementing these strategies has not resulted in elimination of MAP from farms. To date, no study has integrated detailed dairy herd dynamics with different MAP transmission routes. We have developed an individual-based dairy herd model by incorporating basic herd dynamics in a closed herd environment where no new animals have been bought from outside. The model considered three age groups of animals: calves, heifers and adults. It includes sequential life events of a dairy animal and such key dynamic processes of the dairy herd as lactation cycle, calving, voluntary waiting period, insemination, pregnancy, dry-off period and calf and heifer rearing. After initially validating that the model reproduced typical herd dynamics, it was extended by incorporating MAP infection dynamics, where each individual adult animal belonged to one of four infection compartments: susceptible, latent, low shedding and high shedding. The model includes two disease transmission routes: horizontal transmission (i.e., fecal-oral) and vertical transmission (i.e., in utero infection). The results confirm that this model can simulate a realistic dairy herd and that inclusion of the above-mentioned dynamic processes provides useful information about individual infected animals to farmers. Access to the individual animal information offers more validity to assessment of appropriate control strategies for an endemically MAP infected herd. This model can serve as an accurate and novel tool not only to better understand MAP dynamics, but is also valuable as an individual based system of a typical dairy herd that can be applied to other research questions.

The value of pathogen information in treating clinical mastitis.

The objective of this study was to determine the economic value of obtaining timely and more accurate clinical mastitis (CM) test results for optimal treatment of cows. Typically CM is first identified when the farmer observes recognisable outward signs. Further information of whether the pathogen causing CM is Gram-positive, Gram-negative or other (including no growth) can be determined by using on-farm culture methods. The most detailed level of information for mastitis diagnostics is obtainable by sending milk samples for culture to an external laboratory. Knowing the exact pathogen permits the treatment method to be specifically targeted to the causation pathogen, resulting in less discarded milk. The disadvantages are the additional waiting time to receive test results, which delays treating cows, and the cost of the culture test. Net returns per year (NR) for various levels of information were estimated using a dynamic programming model. The Value of Information (VOI) was then calculated as the difference in NR using a specific level of information as compared to more detailed information on the CM causative agent. The highest VOI was observed where the farmer assumed the pathogen causing CM was the one with the highest incidence in the herd and no pathogen specific CM information was obtained. The VOI of pathogen specific information, compared with non-optimal treatment of Staphylococcus aureus where recurrence and spread occurred due to lack of treatment efficacy, was $20.43 when the same incorrect treatment was applied to recurrent cases, and $30.52 when recurrent cases were assumed to be the next highest incidence pathogen and treated accordingly. This indicates that negative consequences associated with choosing the wrong CM treatment can make additional information cost-effective if pathogen identification is assessed at the generic information level and if the pathogen can spread to other cows if not treated appropriately.

Dynamics of Escherichia coli Virulence Factors in Dairy Herds and Farm Environments in a Longitudinal Study in the United States.

Pathogenic Escherichia coli or its associated virulence factors have been frequently detected in dairy cow manure, milk, and dairy farm environments. However, it is unclear what the long-term dynamics of E. coli virulence factors are and which farm compartments act as reservoirs. This study assessed the occurrence and dynamics of four E. coli virulence factors (eae, stx1, stx2, and the gamma allele of the tir gene [γ-tir]) on three U.S. dairy farms. Fecal, manure, water, feed, milk, and milk filter samples were collected from 2004 to 2012. Virulence factors were measured by postenrichment quantitative PCR (qPCR). All factors were detected in most compartments on all farms. Fecal and manure samples showed the highest prevalence, up to 53% for stx and 21% for γ-tir in fecal samples and up to 84% for stx and 44% for γ-tir in manure. Prevalence was low in milk (up to 1.9% for stx and 0.7% for γ-tir). However, 35% of milk filters were positive for stx and 20% were positive for γ-tir. All factors were detected in feed and water. Factor prevalence and levels, expressed as qPCR cycle threshold categories, fluctuated significantly over time, with no clear seasonal signal independent from year-to-year variability. Levels were correlated between fecal and manure samples, and in some cases autocorrelated, but not between manure and milk filters. Shiga toxins were nearly ubiquitous, and 10 to 18% of the lactating cows were potential shedders of E. coli O157 at least once during their time in the herds. E. coli virulence factors appear to persist in many areas of the farms and therefore contribute to transmission dynamics.

Longitudinal data collection of Mycobacterium avium subspecies Paratuberculosis infections in dairy herds: the value of precise field data.

Longitudinal infection data on Mycobacterium avium subspecies paratuberculosis (MAP) was collected on three dairy farms in Northeastern United States during approximately 10 years. Precise data on animal characteristics and animal location within farm were collected on these farms. Cows were followed over time with regard to MAP status during biannual fecal and serum sampling and quarterly serum sampling. Approximately 13 000 serum samples, 6500 fecal samples and 2000 tissue samples were collected during these years. Prevalence of positive samples was 1.4% for serological samples, 2.2% in fecal samples and 16.7% in tissue samples. Infection dynamics of MAP was studied and resulted in a number of potential changes in our understanding of MAP infection dynamics. First, a high prevalence of MAP infection was observed in these herds due to lifetime follow up of cows, including slaughter. Second, two distinctly different infection patterns were observed, so called non-progressors and progressors. Non-progressors were characterized by intermittent and low shedding of MAP bacteria and a virtual absence of a humoral immune response. Progressors were characterized by continuous and progressive shedding and a clearly detectable and progressive humoral immune response. Strain typing of MAP isolates on the three farms identified on two of three farms a dominant strain type, indicating that some strains are more successful in terms of transmission and infection progression. Continuous high quality longitudinal data collection turned out to be an essential tool in our understanding of pathobiology and epidemiology of MAP infections in dairy herds.

The effect of manual and mechanical stimulation on oxytocin release and milking characteristics in Holstein cows milked 3 times daily.

Prestimulation administered to a cow before attachment of the milking unit has historically been performed manually. Development of innovative milking technology has allowed manual stimulation to be replaced by mechanical forms of stimulation. Holstein cows (n=30) were enrolled in a crossover design to determine the effect of manual stimulation (forestripping and drying) and high-vibration pulsation on oxytocin profiles, milk yield, milk flow rates, incidence of delayed milk ejection causing bimodal milk flow curves, and residual milk in Holstein cows milked 3 times daily (3×). All cows were subjected to all treatments. Cows received manual (forestripping and drying) or mechanical (high-vibration pulsation) stimulation along with lag times of 0, 30, or 90 s for 21 consecutive milkings. Forestripping involved the manual removal of 2 streams of milk from each teat and drying of the teats. High-vibration pulsation involved increasing the pulsation cycles from 60 to 300 pulses/min and reducing the vacuum in the pulsation chamber to 20 kPa. The 5 treatments were (1) immediate attachment of the milking machine under normal pulsation (T0); (2) dip plus forestrip and drying with 30-s lag time (FD30); (3) dip plus forestrip and drying with 90-s lag time (FD90); (4) high-vibration pulsation for 30 s (HV30); and (5) high-vibration pulsation for 90 s (HV90). Milk yield per milking averaged 14.0 kg and was significantly different among treatments; however, the maximum difference detected among treatments was 0.8 kg/milking. Milking unit on-time, which represents the time when the milking unit is under normal pulsation and harvesting milk (excluding the high-vibration pulsation time of 30 or 90 s), was shortest (245 s) for cows subjected to 90 s of high-vibration pulsation (HV90) and ranged from 256 to 261 s for all other treatments. Milk harvest may have begun during high-vibration pulsation; however, only 0.13 and 0.32 kg of milk was harvested during high-vibration pulsation for HV30 and HV90, respectively. The incidence of bimodal milk curves was lowest for FD90 (7%) and highest for T0 (21%). The somatic cell count was <72×10(3) cells/mL for all treatments. Residual milk obtained by giving 10 IU of oxytocin in the jugular vein followed by 2 min of milking unit attachment represented 12 to 14% of the total milk and did not differ among treatments. Endogenous oxytocin profiles peaked between 12.4 and 18.3 pg/mL for all treatments, and the peak occurred sooner in manually stimulated cows; however, we detected no difference in oxytocin concentration beyond 2 min after milking unit attachment. High-vibration pulsation elicited a similar oxytocin release when taking the start time of stimulation from manual stimulation or high vibration into consideration. Forestripping for visual observation of milk combined with the use of high-vibration stimulation may reduce variation in the milking routine. A predetermined lag time with minimal variation may be achieved via the time spent in high-vibration stimulation instead of a lag period dictated by milking personnel.

Diversity of Listeria monocytogenes within a U.S. dairy herd, 2004-2010.

Listeria monocytogenes, the causative agent of listeriosis, is frequently isolated from the environment. Dairy cows and dairy farm environments are reservoirs of this pathogen, where fecal shedding contributes to its environmental dispersal and contamination of milk, dairy products, and meat. The molecular diversity of 40 L. monocytogenes isolates representing 3 serogroups (1/2a, 1/2b, and 4b) collected between 2004 and 2010 from the feces of dairy cattle on a single dairy farm was assessed using a multivirulence locus sequence typing (MVLST) assay. The dairy farm L. monocytogenes MVLST patterns were compared to those from 138 strains isolated globally from clinical cases, foods, and the environment. Results of the study demonstrated that several distantly related L. monocytogenes strains persisted among members of the herd over the course of the study while other strains were transient. Furthermore, some strains isolated during this study appear to be distantly related to previously isolated L. monocytogenes while others are closely related to Epidemic Clones associated with human illness. This work demonstrates that dairy cows can be reservoirs of a diverse population of potentially human pathogenic L. monocytogenes that represents a risk to consumers of milk, dairy products, and meat.

Stochastic modeling of imperfect Salmonella vaccines in an adult dairy herd.

Salmonella is a major cause of bacterial foodborne disease. Human salmonellosis results in significant public health concerns and a considerable economic burden. Dairy cattle are recognized as a key source of several Salmonella serovars that are a threat to human health. To lower the risk of Salmonella infection, reduction of Salmonella prevalence in dairy cattle is important. Vaccination as a control measure has been applied for reduction of preharvest Salmonella prevalence on dairy farms. Salmonella vaccines are usually imperfect (i.e., vaccines may provide a partial protection for susceptible animals, reduce the infectiousness and shedding level, shorten the infectious period of infected animals, and/or curb the number of clinical cases), and evaluation of the potential impacts of imperfect Salmonella vaccines at the farm level is valuable to design effective intervention strategies. The objective of this study was to investigate the impact of imperfect Salmonella vaccines on the stochastic transmission dynamics in an adult dairy herd. To this end, we developed a semi-stochastic and individual-based continuous time Markov chain (CTMC) vaccination model with both direct and indirect transmission, and applied the CTMC vaccination model to Salmonella Cerro transmission in an adult dairy herd. Our results show that vaccines shortening the infectious period are most effective in reducing prevalence, and vaccines decreasing host susceptibility are most effective in reducing the outbreak size. Vaccines with multiple moderate efficacies may have the same effectiveness as vaccines with a single high efficacy in reducing prevalence, time to extinction, and outbreak size. Although the environment component has negligible contributions to the prevalence, time to extinction, and outbreak size for Salmonella Cerro in the herd, the relative importance of environment component was not assessed. This study indicates that an effective vaccination program against Salmonella Cerro spread in the herd can be designed with (1) vaccines with a single high efficacy in reducing either the infectious period or susceptibility of the host, or (2) if such single high efficacy vaccines are not available, vaccines with multiple moderate efficacies may be considered instead. These findings are also of general value for designing vaccination program for Salmonella serotypes in livestock.

Different cellular and molecular responses of Bovine milk phagocytes to persistent and transient strains of Streptococcus uberis causing mastitis.

Streptococcus uberis is frequently isolated from milk collected from dairy cows with mastitis. According to the host's immunity, bacterial virulence, and their interaction, infection with some strains can induce persistent subclinical inflammation, while infection with others induces severe inflammation and transient mastitis. This study compared the inflammatory response of milk-isolated white blood cells (mWBCs) to persistent and transient S. uberis strains. Quarter milk samples were collected aseptically for bacterial culture from all lactating cows once a week over a 10-week period. A transient and noncapsular strain with a 1-week intramammary infection duration was selected from this herd, while a persistent and capsular S. uberis strain with an intramammary infection longer than 2 months from our previous study was selected based on an identical pulse field gel electrophoresis pattern during the IMI episode. Cellular and molecular responses of mWBCs were tested, and the data were analyzed using repeated analysis of variance. The results showed a higher response in migration, reactive oxygen species generation, and bacterial killing when cells were stimulated with transient S. uberis. In contrast, the persistent strain led to increased neutrophil extracellular trap release. This study also highlighted several important molecular aspects of mWBCs. Gene expression analyses by real-time RT-PCR revealed a significant elevation in the expression of Toll-like receptors (TLR-1, TLR-2, TLR-6) and proinflammatory cytokines (tumor necrosis factor-alpha or TNF-α) with the transient strain. Additionally, Streptococcus uberis capsule formation might contribute to the capability of these strains to induce different immune responses. Altogether, these results focus on the immune function of activated mWBCs which demonstrate that a transient strain can elicit a stronger local immune response and, subsequently, lead to rapid recovery from mastitis.

Effect of lactation therapy on Staphylococcus aureus transmission dynamics in two commercial dairy herds.

BACKGROUND: Treatment of subclinical mastitis during lactation can have both direct (individual animal level) and indirect (population level) effects. With a few exceptions, prior research has focused on evaluating the direct effects of mastitis treatment, and to date no controlled field trials have been conducted to test whether beneficial indirect effects of lactation treatment strategies targeting subclinical mastitis can be demonstrated on commercial dairy farms. Furthermore, there is limited knowledge on the impact of such interventions on the population dynamics of specific bacterial strains. The purpose of this study was to test the hypothesis that lactation therapy targeting S. aureus subclinical intramammary infection reduces transmission of S. aureus strains within dairy herds. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to determine strain specific infection dynamics in treated and control groups in a split herd trial conducted on 2 commercial dairy farms. RESULTS: The direct effect of 8 days intramammary lactation therapy with pirlimycin hydrochloride was demonstrated by an increased proportion of cure and a reduction in duration of infection in quarters receiving treatment compared to untreated controls. The indirect effect of lactation therapy was demonstrated by reduction of new S. aureus intramammary infections (IMI) caused by the dominant strain type in both herds. Strain typing of representative isolates taken over the duration of all IMI, including pre- and post-treatment isolates, provided more precise estimates of new infection, cure, and re-infection rates. New S. aureus infections in recovered susceptible quarters and the emergence of a new strain type in one herd influenced incidence measures. CONCLUSION: In addition to demonstrating positive direct effects of lactation therapy, this study provides evidence that treatment of subclinical S. aureus mastitis during lactation can have indirect effects including preventing new IMI and reducing incidence of clinical mastitis within dairy herds. Strain specific transmission parameter estimates for S. aureus MLST clonal complexes 5, 97 and 705 in 2 commercial dairy herds are also reported.

Associations between Mycobacterium avium subsp. paratuberculosis antibodies in bulk tank milk, season of sampling and protocols for managing infected cows.

BACKGROUND: The objective of this study was to identify associations between the concentration of Mycobacterium avium subsp. paratuberculosis (MAP) antibodies in bulk milk and potential risk factors in herd management and herd characteristics, explaining high MAP antibody titers in milk. An extensive questionnaire was administered to 292 organic and conventional dairy farms from New York, Wisconsin and Oregon. Bulk milk samples were taken from each farm for MAP enzyme-linked immunosorbent assay (ELISA). A general linear model was constructed with MAP ELISA value as the outcome variable and the management factors and herd characteristics as independent variables, while at the same time controlling for the study design variables of state, herd size, and production system (organic or conventional). High bulk tank MAP ELISA value may be due to either a high prevalence of MAP in a herd with many cows contributing to the antibody titer or due to a few infected cows that produce large quantities of antibodies. RESULTS: Results of the regression models indicated that bulk milk ELISA value was associated with season of sampling and the presence or absence of protocols for managing MAP-positive cows. The concentration of MAP antibodies in bulk milk varied seasonally with a peak in the summer and low concentrations in the winter months. When compared to farms that had never observed clinical Johne's disease, keeping MAP-positive cows or only culling them after a period of delay was associated with an increase in optical density. CONCLUSIONS: The seasonal variation in MAP antibody titers, with a peak in the summer, may be due to a seasonal increase in MAP-bacterial load. Additionally, seasonal calving practices may contribute to seasonal fluctuations in MAP antibody titers in bulk tank milk. Keeping MAP-positive cows increases the antibody titer in bulk milk, likely due to direct antibody production in the infected cow and indirect triggering of antibody production in herdmates.

Invasion and transmission of Salmonella Kentucky in an adult dairy herd using approximate Bayesian computation.

BACKGROUND: An outbreak of Salmonella Kentucky followed by a high level of sustained endemic prevalence was recently observed in a US adult dairy herd enrolled in a longitudinal study involving intensive fecal sampling. To understand the invasion ability and transmission dynamics of Salmonella Kentucky in dairy cattle, accurate estimation of the key epidemiological parameters from longitudinal field data is necessary. The approximate Bayesian computation technique was applied for estimating the transmission rate (ß), the recovery rate (γ) and shape (n) parameters of the gamma distribution for the infectious (shedding) period, and the basic reproduction ratio (R0), given a susceptible-infectious-recovered-susceptible (SIRS) compartment model with a gamma distribution for the infectious period. RESULTS: The results report that the mean transmission rate (ß) is 0.417 month-1 (median: 0.417, 95% credible interval [0.406, 0.429]), the average infectious period (γ-1) is 7.95 months (median: 7.95, 95% credible interval [7.70, 8.22]), the mean shape parameter (n) of the gamma distribution for the infectious period is 242 (median: 182, 95% credible interval [16, 482]), and the mean basic reproduction ratio (R0) is 2.91 (median: 2.91, 95% credible interval [2.83, 3.00]). CONCLUSIONS: This study shows that Salmonella Kentucky in this herd was of mild infectiousness and had a long infectious period, which together provide an explanation for the observed prevalence pattern after invasion. The transmission rate and the recovery rate parameters are inferred with better accuracy than the shape parameter, therefore these two parameters are more sensitive to the model and the observed data. The estimated shape parameter (n) has large variability with a minimal value greater than one, indicating that the infectious period of Salmonella Kentucky in dairy cattle does not follow the conventionally assumed exponential distribution.

Importance of differential expression of Marek's disease virus gene pp38 for the pathogenesis of Marek's disease.

The role of pp38 in the pathogenesis of Marek's disease (MD) has not been fully elucidated. Previously, we reported the presence of two splice variants (Spl A and Spl B) for pp38. We also reported that the wild-type pp38 (WT), as well as the mutated pp38 (MUT), altered the oxidative phosphorylation pathway in infected cells. To determine whether the different forms of pp38 are important for the pathogenesis of MD, we generated RB-1B-based bacterial artificial chromosome (BAC) clones expressing pp38MUT, pp38Sp1 A, and pp38Spl B. Infectious viruses were recovered from these BAC clones in chick kidney cells (CKC). The Spl A and Spl B viruses had significantly smaller plaque sizes and replicated to a lesser degree in CKC than the WT and MUT viruses. Two in vivo experiments were conducted by inoculating 7-day-old P2a chicks with 1000 plaque-forming units of each virus. In the first experiment, chicks were sacrificed at 4, 8, 11, and 15 days postinfection (PI). WT and MUT viruses had similar viremia levels using virus isolation and quantitative real-time PCR (qPCR) assays, whereas Spl A and Spl B viruses had significantly lower viremia levels than WT and MUT viruses. In the second experiment, we showed that tumor development and MD mortality were similar in the WT- and MUT-infected chickens, with all birds MD positive at 5 wk PI. In contrast, chickens infected with Spl B and Spl A had a significantly lower MD incidence at 11 wk PI, when the experiment was terminated.

Evaluation of an alternative dosing regimen of a J-5 mastitis vaccine against intramammary Escherichia coli challenge in nonlactating late-gestation dairy cows.

The objective of the study was to evaluate the efficacy of an alternative vaccination regimen of a J-5 bacterin against intramammary Escherichia coli challenge in nonlactating late-gestation dairy cows. The parameters analyzed to assess the effect of vaccination were milk yield, milk conductivity, somatic cell count, J-5-specific serum IgG titers, and clinical signs. Twenty multiparous Holstein cows from the Cornell teaching and research dairy herd were paired by days in milk and were randomly selected to receive either the alternative off-label regimen of commercial J-5 bacterin or act as nonvaccinated controls. Cows received a first dose of bacterin 15 d before dry off, a second dose with the same product at the day of dry off, and the third dose 2 wk after dry off. The cows in both groups were challenged 10 d before the expected calving date. Serum IgG (total, IgG1 and IgG2) levels were higher in vaccinates compared with control cows. Eighty-five percent of challenged quarters became infected between both groups of animals. Eight of the 10 vaccinated and 9 of the 10 control cows showed signs of clinical mastitis postfreshening. A non-severe clinical mastitis was observed 24 to 48 h postparturition, characterized by flakes or clots in milk and mild swelling or pain. Off-label vaccination did reduce the clinical severity of clinical mastitis in the vaccinated group of cows as evidenced by reduced California mastitis test score, fewer flakes and lower overall clinical mastitis score. No significant differences between vaccinated and control groups were detected for rectal temperature. In conclusion, the alternative off-label vaccination scheme used in our study and evaluated in a novel E. coli challenge model did not prevent new intramammary infections but reduced clinical severity of experimentally induced E. coli mastitis.

Association of pathogen-specific clinical mastitis in the first 100 days of first lactation with productive lifetime: An observational study comparing competing risks models for death and sale with the Cox model.

The objective of this observational study was to study the association between clinical mastitis (CM) (Streptococcus spp., Staphylococcus aureus, Staphylococcus spp., Escherichia coli, Klebsiella spp., cases with other treated or other not treated organisms, CM without growth) occurring in a dairy cow's first 100 days (d) of her first lactation and her total productive lifetime, ending in death or sale (for slaughter). Data were collected from 24,831 cows in 5 New York Holstein herds from 2004 to 2014. Two analytical approaches were compared. First, removals (death, sale) were treated as competing events in separate survival analyses, in proportional subdistribution hazards models. In one, death was coded as the event of interest and sale as the competing event; in another, sale was the event of interest and death the competing event. Second, traditional survival analysis (Cox proportional hazards) was conducted. In all models, the time variable was number of days from date of first calving until event (death or sale) date; if the cow was alive at study end, she was censored. Models were stratified by herd. Ten percent of cows died; 48.4 % were sold. In the competing risks analysis, E. coli and CM without growth were associated with death; the former with an increased hazard rate of death, the latter with a lower one. Streptococcus spp., Staph. aureus, Klebsiella spp., cases with other treated or untreated organisms, and CM without growth were associated with higher hazard rates of sale. The Cox proportional hazards model's hazard rates were higher than those in the competing risks model in which death was the event of interest, and resembled those in the model in which sale was the event of interest. Four additional Cox models, omitting dead or sold cows, or censoring each, were also fitted; hazard ratios were similar to the above models. Proportional subdistribution hazards models were appropriate due to competing risks (death, sale); they produce less-biased estimates. A study limitation is that while proportional subdistribution hazards models were appropriate, they have the illogical feature of keeping subjects at risk for the event of interest even after experiencing the competing event. This is, however, necessary in estimating cumulative incidence functions. Another limitation concerns pathogen variability among study farms, implying that CM decisions are farm-specific. Misclassification of 'dead' vs. 'sold' cows was also possible. Nevertheless, the findings may help in optimizing management of cows contracting specific types of CM early in productive lifetime.