The dynamics of Staphylococcus aureus intramammary infection in nine Danish dairy herds

The aim of the present study was to examine the diversity of Staphylococcus aureus isolates from bovine intramammary infections (IMI) in nine dairy herds, and compare these with isolates from other sites on the cows by phage- and ribotyping. Whether colonisation of milkers with S. aureus could be a source of infection for bovine IMI was investigated. In addition, 100 epidemiologically unrelated S. aureus isolates from asymptomatic human carriers were also phage- and ribotyped to compare the human and bovine reservoir of S. aureus in Denmark. A total of 625 S. aureus isolates from bovine IMI, bovine skin lesions, milking personnel, and non-farm-related human carriers were included in the study. Certain types predominated in one or several herds during the study period of one-and-a-half to two years, whereas the presence of other types was of a more sporadic nature. Within the individual herds, there was a close correspondence between ribo- and phage types of S. aureus isolated from bovine intramammary infections and skin lesions. Isolates from milking personnel, however, were not identical to any of the predominant intramammary strains. Furthermore, several of the isolates from milking personnel showed ribo- and phage patterns identical to S. aureus isolates from human carriers. The findings of the present study underline the importance of strict milking hygiene and improvement of current mastitis therapy. The results support the hypothesis that some S. aureus mastitis strains are more contagious, virulent or persistent than others. The human reservoir of S. aureus does not play a major role as a source of bovine intramammary infections.     

Loser cows in Danish dairy herds: risk factors

The objective of our study was to evaluate risk factors for the loser cow state in Danish dairy herds. A loser cow was defined on the basis of a clinical examination of the individual cow. Recordings of clinical signs were converted into a loser cow score and all cows with a loser cow score of 8 or more were defined as loser cows. We used correspondence analysis to give a first indication of the associations between the proportion of loser cows in the herd and potential risk factors. Risk factors were evaluated both at the herd level (39 herds) and at the cow level (6451 cows) using logistic regression. Conventional herds with a high average somatic cell count, hard cubicles and no grazing seem to be associated with a high proportion of loser cows. Odds ratio for the loser cow state was 4.18 in conventional herds compared to organic herds. Odds ratio for the loser cow state was 1.87 for an increase in average weighted mean bulk tank somatic cell count of 100,000 cells per ml. We found a statistically significant interaction between stall surface and the use of grazing. Odds ratio for the loser cow state in herds with soft cubicles and the use of grazing was 0.09 compared to herds with hard cubicles and no grazing. At the cow level, odds ratio for the loser cow state increases significantly with increasing parity. Odds ratios for the loser cow state were 1, 2.59 and 6.74 for parity 1, 2, and 3 or older, respectively. Odds ratio for the loser cow state was significantly higher (3.38) for cows giving birth to twins at the last calving before the scoring. Additionally, cows scored on pasture had a significantly lower odds ratio (0.26) for the loser cow state compared to cows scored in the barn.     

Prevalence of Coxiella burnetii antibodies in Danish dairy herds

During recent years in Denmark higher rates of antibodies to Coxiella burnetii have been detected in animals and humans than previously reported. A study based on bulk tank milk samples from 100 randomly selected dairy herds was performed to estimate the prevalence and geographical distribution of antibody positive dairy herds. Using the CHEKIT Q-Fever Antibody ELISA Test Kit (IDEXX), the study demonstrated a prevalence of 59% antibody positive herds, 11% antibody intermediate herds and 30% antibody negative herds based on the instructions provided by the manufacturer. The geographical distribution does not indicate a relationship between the regional density of dairy farms and the prevalence of antibody positive dairy farms. The result supports the hypothesis of an increase in the prevalence of positive dairy herds compared to previous years.     

Increasing prevalence of Coxiella burnetii seropositive Danish dairy cattle herds

A study based on bulk tank milk samples from 120 randomly selected dairy cattle herds was conducted to estimate the prevalence of Coxiella burnetii seropositive dairy herds, to describe the geographical distribution, and to identify risk factors. Using the CHEKIT Q-fever Antibody ELISA Test Kit (IDEXX), the study revealed a prevalence of 79.2% seropositive herds, 18.3% seronegative herds, and 2.5% serointermediate herds based on the instructions provided by the manufacturer. Multifactorial logistic regression showed statistically significant associations (P < 0.01) between C. burnetii seropositivity and increasing herd size (OR = 1.02 per cow increment) and increasing regional average number of cattle per dairy herd (OR = 1.02 per animal increment). Herds >150 cows had 17.9 times higher odds of testing positive compared to herds <80 cows. The regional average number of cattle herds per square kilometer was borderline significantly related to the occurrence of seropositive dairy herds (P = 0.06). The results indicate an increased prevalence of seropositive dairy herds since the previous survey in 2008 and an adverse impact of increasing herd size and cattle density on the risk of seropositivity.     

The effect of grazing on cow mortality in Danish dairy herds

The effect of summer grazing in large Danish dairy herds and certain management characteristics of grazing were studied for their impact on dairy cow mortality. Mortality data (from the Danish Cattle Database) from 391 Danish dairy herds (>100 cows) were combined with information from a questionnaire survey of grazing procedures on these herds in 2008. In all, 131 of the herds were identified as summer grazing and 260 as zero-grazing herds. The mortality was affected by an interaction of summer grazing and milking system. The risk of a cow dying was reduced to 46% in a grazing compared to a zero-grazing herd having automatic milking system. In traditional milking system, mortality was reduced to 75% in grazing compared to zero-grazing herds. Within the grazing herds, the risk of mortality decreased with increasing number of hours on pasture during the season. Free access between barn and pasture was associated with increased cow mortality.     

Managerial and environmental determinants of clinical mastitis in Danish dairy herds

Background

Several management and environmental factors are known as contributory causes of clinical mastitis in dairy herd. The study objectives were to describe the structure of herd-specific mastitis management and environmental factors and to assess the relevance of these herd-specific indicators to mastitis incidence rate.

Methods

Disease reports from the Danish Cattle Data Base and a management questionnaire from 2,146 herds in three Danish regions were analyzed to identify and characterize risk factors of clinical mastitis. A total of 94 (18 continuous and 76 discrete) management and production variables were screened in separate bivariate regression models. Variables associated with mastitis incidence rate at a p-value < 0.10 were examined with a factor analysis to assess the construct of data. Separately, a multivariable regression model was used to estimate the association of management variables with herd mastitis rate.

Results

Three latent factors (quality of labor, region of Denmark and claw trimming, and quality of outdoor holding area) were identified from 14 variables. Daily milk production per cow, claw disease, quality of labor and region of Denmark were found to be significantly associated with mastitis incidence rate. A common multiple regression analysis with backward and forward selection procedures indicated there were 9 herd-specific risk factors.

Conclusion

Though risk factors ascertained by farmer-completed surveys explained a small percentage of the among-herd variability in crude herd-specific mastitis rates, the study suggested that farmer attitudes toward mastitis and lameness treatment were important determinants for mastitis incidence rate. Our factor analysis identified one significant latent factor, which was related to labor quality on the farm.     

Clinical parameters for assessment of udder health in Danish dairy herds

This study examined the possibilities of using clinical parameters related to the bovine udder for characterisation of udder health. Five clinicians performed systematic clinical recordings of udder health at 3 visits to 4 dairy herds. Several of the clinical parameters were scored on an ordinal scale. The agreement between clinicians was compared using kappa statistics. Factor analysis was used to identify udder types. The clinical evaluations showed substantial variation among clinicians. Parameters that were not directly related to pathological conditions showed the highest variation e.g. length of the claws, teat shape and hardness of the udder parenchyma. On the other hand, evaluation of pathological parameters such as nodes in the udder, skin lesions and oedema showed good agreement between clinicians. Udder types identified by means of factor analysis were found to be suitable for characterisation of udder health. Especially one factor related to dry quarters and udder asymmetry showed a more consistent relationship to milk yield than traditionally applied udder health parameters such as treatment rate and cell count. It is concluded that there is a considerable need for increased efforts among clinicians in order to standardise clinical recordings. It is further concluded that certain combinations of extended clinical recordings have significant perspectives for future characterisation of udder health.     

Loser cows in Danish dairy herds: definition, prevalence and consequences

During the last few years, many Danish dairy farmers have expressed increasing concerns regarding a group of cows, which we have chosen to term ‘loser cows’. Until now, a loser cow has not been described scientifically. We defined a loser cow on the basis of a clinical examination of the cow. A total of 15,151 clinical examinations were made on 6,451 individual cows from 39 randomly selected, large Danish dairy herds with loose-housing systems using a clinical protocol. Scores for the clinical signs lameness, body condition, hock lesions, other cutaneous lesions, vaginal discharge, condition of hair coat and general condition were converted into a loser cow score. Cows with a loser cow score of 8 or more were classified as loser cows. The overall prevalence of loser cows was 2.15%, 4.50% and 2.98% during the first, second and third round of herd visits, respectively.

The associations between the loser cow state and milk production, mortality, morbidity, culling and workload for the farmer were evaluated using data from herd visits and from the Danish Cattle Database and a number of different statistical techniques. It was concluded that the loser cow state has significant negative consequences for both the farmer and the cow. On average, loser cows yielded 0.61 to 2.24 kg energy corrected milk less per day than non-loser cows depending on parity. Hazard ratio for death or euthanasia was 5.69 for loser cows compared to non-loser cows. Incidence rate ratio for disease treatments was 0.69 for non-loser cows compared to loser cows. Loser cows were often culled in an ‘unfavourable’ way and generally caused extra workload for the farmer.

A simplified version of the loser cow score was evaluated and is recommended for future research and use in practice.     

Herd-level risk factors for the mortality of cows in Danish dairy herds

The factors affecting the mortality of cows in Danish dairy herds were investigated by analysing data from 6839 herds. The mean risk of mortality during the first 100 days of lactation was 2.5 per cent. The risk of mortality increased with increasing herd size (odds ratio [OR] 1.05 for an increase in herd size of 50 cows), with the proportion of purchased cows (OR 1.05 for an increase in the proportion of purchased cows of 0.1), and with increases in the somatic cell count (OR 1.16 for an increase in average weighted mean somatic cell count of 100,000 cells/ml). The risk decreased with increasing average milk yield per cow (OR 0.93 for an increase in mean yield per cow-year of 1000 kg). The risk was lower in free-stall barns with deep litter (OR 0.79) than in barns with cubicles (OR 1) or tie-stalls (OR 1.04). Herds with Danish Holstein (OR 1) or Danish Jersey (OR 0.93) cows as the predominant breed had a higher risk of mortality than herds of the Danish red dairy breed (OR 0.67). The risk of mortality was lower in organic herds (OR 1) than in conventional herds (OR 1.17), and lower in herds that grazed pasture during the summer (OR 0.78).     

Extent of Mycobacterium bovis infection in dairy cattle herds subject to partial culling as determined by an interferon-gamma assay

The interferon-gamma (IFN-γ) assay is employed as a complementary diagnostic test for bovine tuberculosis (BTB) in many countries. To simplify this assay, we established a 96-well plate format using the ESAT-6 and CFP-10 antigens and then employed it to determine the extent of Mycobacterium (M.) bovis infection in dairy herds with a history of BTB outbreaks in a country where only selective culling is practiced. The sensitivity and specificity of this IFN-γ assay were 85.9% and 100%, respectively, based on comparison with the conventional single intradermal tuberculin test (SIDT). The IFN-γ assay was also positive in 30.4% and 36.8% of SIDT-negative animals from herds with recent and remote BTB outbreaks, respectively. Of 14 SIDT-negative, IFN-γ positive cattle, five (35.7%) were culture positive and an additional six were positive based on a polymerase chain reaction-based test for M. bovis. Therefore, the IFN-γ assay has the potential to serve as a specific and sensitive test for M. bovis infection in dairy cattle. Further, the results indicated that a substantial portion of SIDT-negative animals in herds with previous BTB outbreaks were actually infected with M. bovis. Accordingly, the present selective-culling strategy may require modifications to include this more sensitive assay.     

Testing of bulk tank milk for Salmonella Dublin infection in Danish dairy herds.

The usefulness of enzyme-linked immunosorbent assay (ELISA) was investigated as a simple method to screen for Salmonella Dublin infection in dairy herds, examining bulk tank milk samples for lipopolysaccharide (O:1,9,12) antibodies. The cut-off value for the ELISA on bulk tank milk was established based on individual milk samples (n = 2887) and bulk tank milk from 52 herds. Bulk tank milk samples (n = 5108) were collected from 1464 dairy herds located in 19 different areas. About 10% of the dairy herds in Denmark participated in the study. The percentage of herds changing from test-negative to test-positive in each area was correlated with the incidence of S. Dublin outbreaks in the corresponding county (r = 0.48, n = 19; P < 0.025). The mean level of the OD values obtained in the first and third test rounds was not constant (Pr /t/ = 0.0001). The study demonstrated that the probability of being test-negative in the third test round was 0.926 for a herd with 2 previous test-negative results. It was concluded that the investigated ELISA method was in general accordance with the cases of clinical S. Dublin infection recorded, and that the method has a potential for national screening purposes.     

Risk factors for Coxiella burnetii antibodies in bulk tank milk from Danish dairy herds

The aim was to identify risk factors associated with Coxiella burnetii antibody positivity in bulk tank milk (BTM) samples from 100 randomly selected Danish dairy cattle herds. Antibody levels were measured by an enzyme-linked immuno-sorbent assay. Before testing the herds, the farm managers were interviewed about hired labour, biosecurity, housing and herd health during the 12 months prior to the study. Variables considered important for C. burnetii antibody positivity in multivariable logistic regression analysis included the sharing of machines between farms (OR = 3.6), human contacts (OR = 4.2), artificial insemination by other people than artificial insemination technicians (OR = 7.7), routine herd health contract with the veterinarian (OR = 4.3) and hygiene precautions taken by veterinarians (OR = 5). In addition, herd size, hired labour, trading of cattle between farms, quarantine and use of calving and disease pens also showed significant association in univariable analysis. This study demonstrates that strict biosecurity is important for the prevention of infections with C. burnetii.     

Annual incidence, prevalence and transmission characteristics of Streptococcus agalactiae in Danish dairy herds

Contagious mastitis pathogens continue to pose an economic threat to the dairy industry. An understanding of their frequency and transmission dynamics is central to evaluating the effectiveness of control programmes. The objectives of this study were twofold: (1) to estimate the annual herd-level incidence rates and apparent prevalences of Streptococcus agalactiae (S. agalactiae) in the population of Danish dairy cattle herds over a 10-year period from 2000 to 2009 inclusive and (2) to estimate the herd-level entry and exit rates (demographic parameters), the transmission parameter, β, and recovery rate for S. agalactiae infection. Data covering the specified period, on bacteriological culture of all bulk tank milk samples collected annually as part of the mandatory Danish S. agalactiae surveillance scheme, were extracted from the Danish Cattle Database and subsequently analysed. There was an increasing trend in both the incidence and prevalence of S. agalactiae over the study period. Per 100 herd-years the value of β was 54.1 (95% confidence interval [CI] 46.0-63.7); entry rate 0.3 (95% CI 0.2-0.4); infection-related exit rate 7.1 (95% CI 5.6-8.9); non-infection related exit rate 9.2 (95% CI 7.4-11.5) and recovery rate 40.0 (95% CI 36.8-43.5). This study demonstrates a need to tighten the current controls against S. agalactiae in order to lower its incidence.     

Bulk-tank milk ELISA antibodies for estimating the prevalence of paratuberculosis in Danish dairy herds

Paratuberculosis (Johne's disease) has been widespread in Danish dairy herds for a long time but the herd-level prevalence has never been determined precisely. To evaluate the prevalence of paratuberculosis in Danish dairy herds in various regions, an ELISA based on a commercially available antigen was adapted for testing bulk-tank milk for the presence of antibodies to Mycobacterium avium subsp. paratuberculosis. Bulk-tank milk samples were collected from six milk-collecting centres from six different areas of the country. Samples from 900 herds (about 7.5% of all Danish dairy herds) were examined, and 70% were positive at the statistically optimal cut-off (sensitivity 97.1%; specificity 83.3%). The technical performance of the ELISA was not sufficient to provide a tool for surveillance because even slight changes in optical density for the samples would change the classification of some samples. The infection is more widespread than previous investigations have shown.     

Spatial differences in occurrence of paratuberculosis in Danish dairy herds and in control programme participation

Paratuberculosis is a chronic infection of economic importance to the cattle industry and a voluntary control programme is offered to Danish dairy farmers. Our objective was to evaluate spatial differences in both control programme participation and paratuberculosis prevalence in Denmark. The study included 4414 dairy herds: 1249 were participating in the control programme, and 1503 were tested for antibodies to Mycobacterium avium subsp. paratuberculosis (MAP). Spatial differences were evaluated by kernel smoothing, kriging, and cluster analysis. Participation was lowest among herds on the island Zealand (≤23%). The risk of a herd being infected with MAP was found to be high on most of Zealand, but the uncertainty of this result was large due to a limited number of tested herds. In the rest of the country, the south western part of the peninsula Jutland had the highest risk of MAP (≥91%). The risk of MAP was also high (86-91%) in the northern part of both Jutland and Funen. The predicted apparent within-herd prevalence was highest (5-8.5%) in some local areas across Jutland, in the north western part of the island Funen, and in the south and western part of Zealand. Scan statistics located the primary cluster of herds with high apparent within-herd prevalence in the western part of Funen. Furthermore, a number of significant clusters were found in Jutland and a single significant cluster in Zealand. Consistency was found between kriging and scan statistics results with respect to location of areas with high apparent within-herd prevalence of MAP. Potential explanations for differences in participation include herd size and local herd health advisers, whereas for example soil characteristics might influence prevalence. Further studies are needed to evaluate these and other risk factors.     

Prevalence of Michigan dairy herds infected with Mycobacterium avium subspecies paratuberculosis as determined by environmental sampling

A cross-sectional, stratified random survey of Michigan dairy herds was conducted to estimate the prevalence of herds infected with Mycobacterium avium paratuberculosis (MAP), the causative agent of Johne's disease, in Michigan using targeted environmental sampling. One pooled sample each from the primary manure storage area and a high-traffic common cow area from each herd was collected and cultured for MAP using the ESP^® culture system II. A herd was classified as positive if at least one sample was culture positive for MAP. State, agricultural district, and herd size stratum prevalence were calculated. Information on past MAP testing and cattle purchase history was collected, and logistic regression was performed to determine their importance to the MAP status of the herd. One hundred twenty-seven herds were contacted, and 94 agreed to participate in the study. The environment of 38 (40.4%) herds cultured positive for MAP. MAP was found in all herds (n = 15) with greater than 200 lactating cows. Herds that had tested for MAP or purchased cattle in the previous 5 years were 4.6 and 3.1 times, respectively, more likely to be infected than herds that had not. MAP continues to be prevalent on Michigan dairy farms, especially those with greater than 200 lactating cows. The environmental sampling protocol used in this study is an economically attractive alternative for monitoring herd level prevalence and the progress of Johne's disease control programs at the state or national level. Implementation of such a program would aid states in monitoring Johne's control program progress, and guide changes over time.     

Evaluation of test-strategies for estimating probability of low prevalence of paratuberculosis in Danish dairy herds

Paratuberculosis is a chronic infection affecting cattle and other ruminants. In the dairy industry, losses due to paratuberculosis can be substantial in infected herds and several countries have implemented national programmes based on herd-classification to manage the disease. The aim of this study was to develop a method to estimate the probability of low within-herd prevalence of paratuberculosis for Danish dairy herds. A stochastic simulation model was developed using the R^® programming environment. Features of this model included: use of age-specific estimates of test-sensitivity and specificity; use of a distribution of observed values (rather than a fixed, low value) for design prevalence; and estimates of the probability of low prevalence (Pr_Low) based on a specific number of test-positive animals, rather than for a result less than or equal to a specified cut-point number of reactors.

Using this model, five herd-testing strategies were evaluated: (1) milk-ELISA on all lactating cows; (2) milk-ELISA on lactating cows ≤4 years old; (3) milk-ELISA on lactating cows >4 years old; (4) faecal culture on all lactating cows; and (5) milk-ELISA plus faecal culture in series on all lactating cows.

The five testing strategies were evaluated using observed milk-ELISA results from 19 Danish dairy herds as well as for simulated results from the same herds assuming that they were uninfected.

Whole-herd milk-ELISA was the preferred strategy, and considered the most cost-effective strategy of the five alternatives. The five strategies were all efficient in detecting infection, i.e. estimating a low Pr_Low in infected herds, however, Pr_Low estimates for milk-ELISA on age-cohorts were too low in simulated uninfected herds and the strategies involving faecal culture were too expensive to be of practical interest. For simulated uninfected herds, whole-herd milk-ELISA resulted in median Pr_Low values >0.9 for most herds, depending on herd size and age-structure. None of the strategies provided enough power to establish a high Pr_Low in smaller herds, or herds with a younger age-structure. Despite this, it appears as if the method is a useful approach for herd-classification for most herds in the Danish dairy industry.     

Associations of Neospora caninum seropositivity with gestation number and pregnancy outcome in Danish dairy herds.

The prevalence and distribution of seropositivity towards the protozoan parasite Neospora caninum were studied in single blood samples from 1561 cows from 31 Danish dairy herds. Blood samples were analysed by an indirect enzyme-linked immunoassay and an indirect fluorescent-antibody test. Seroprevalence in 15 herds with previous abortions assigned to neosporosis ranged from 1% to 58%, with a mean frequency of 22%. In eight out of 16 herds without a history of N. caninum related abortions, no seroreactors were found. In the remaining eight herds, the seroprevalence ranged from 6% to 59%. The prevalence and distribution of seropositivity, gestation number prior to sampling, and breed were related to abortions and perinatal deaths using a random-effects logistic-regression model. Abortion risk was significantly increased in seropositive animals (OR=3) and in 2nd-gestation cows (OR=3). Perinatal death was significantly influenced by gestation number and breed, but not by serostatus. Reproductive performance and culling risk of cows were not affected by serostatus. Seropositivity increased with “age” (i.e. gestation number) (P=0.02). In open cows, seropositivity tended to decrease with distance from calving (P=0.05). The proportion of seropositive pregnant cows increased with trimester (P=0.02).