High Within-herd Prevalence Research Articles

Prevalence of paratuberculosis in Thuringian sheep and goat flocks

In Germany, only few data on the current distribution of paratuberculosis in sheep and goat flocks is available. The present study provides an overview of the distribution of Mycobacterium avium ssp. paratuberculosis (MAP) in 165 Thuringian sheep and goat flocks. Also, the study investigated the association between the MAP status of the flock and herd specific factors as well as the association between the individual measured value of ELISA and animal specific factors like age, body condition, sex, and animal species. To investigate the prevalence of MAP, serum samples from 2550 sheep and 1171 goats from 165 flocks (flock size 2 to 2879 animals) were serologically examined for MAP antibodies in 2021. Additionally, 1 to 6 environmental faecal samples were collected from every flock depending on the flock size. They were examined for the presence of MAP by using both bacteriological cultivation and a commercially available real-time-PCR. MAP antibodies were detected in 41 sheep (1.6%) and 29 goats (2.5%), which accounts to a detection of MAP antibodies in 20.6% of the 165 flocks (on herd level). The symptoms of paratuberculosis, weight loss with preserved appetite and altered fecal consistency, were observed in only four of the flocks. A positive association was identified between the detection of MAP or MAP-specific antibodies in a flock and flock size, as well as positive association between the measured value in the Elisa (s/p ratio) and the age of the animal. Furthermore, an association between an increasing s/p ratio of the ELISA and a decreasing body condition was found. Given what is known about the distribution of paratuberculosis in small ruminants, this disease should always be considered as a possible cause of weight loss and diarrhea. In case of high within-herd prevalence herd-specific control measures should be considered. In serological herd monitoring, animals with poor body condition should preferably be included in the sample, as the probability of being able to identify MAP positive animals is higher here.

The secretome of Staphylococcus aureus strains with opposite within-herd epidemiological behavior affects bovine mononuclear cell response

Staphylococcus aureus modulates the host immune response directly by interacting with the immune cells or indirectly by secreting molecules (secretome). Relevant differences in virulence mechanisms have been reported for the secretome produced by different S. aureus strains. The present study investigated the S. aureus secretome impact on peripheral bovine mononuclear cells (PBMCs) by comparing two S. aureus strains with opposite epidemiological behavior, the genotype B (GTB)/sequence type (ST) 8, associated with a high within-herd prevalence, and GTS/ST398, associated with a low within-herd prevalence. PBMCs were incubated with different concentrations (0%, 0.5%, 1%, and 2.5%) of GTB/ST8 and GTS/ST398 secretome for 18 and 48 h, and the viability was assessed. The mRNA levels of pro- (IL1-β and STAT1) and anti-inflammatory (IL-10, STAT6, and TGF-β) genes, and the amount of pro- (miR-155-5p and miR-125b-5p) and anti-inflammatory (miR-146a and miR-145) miRNAs were quantified by RT-qPCR. Results showed that incubation with 2.5% of GTB/ST8 secretome increased the viability of cells. In contrast, incubation with the GTS/ST398 secretome strongly decreased cell viability, preventing any further assays. The GTB/ST8 secretome promoted PBMC polarization towards the pro-inflammatory phenotype inducing the overexpression of IL1-β, STAT1 and miR-155-5p, while the expression of genes involved in the anti-inflammatory response was not affected. In conclusion, the challenge of PBMC to the GTS/ST398 secretome strongly impaired cell viability, while exposure to the GTB/ST8 secretome increased cell viability and enhanced a pro-inflammatory response, further highlighting the different effects exerted on host cells by S. aureus strains with epidemiologically divergent behaviors.

Bayesian hierarchical modelling of the geospatial distribution of fasciolosis in dairy cattle and the impact on production: Application to the main milk-producing region (Galicia) in Spain

Fasciolosis caused by Fasciola hepatica is a common parasitic infection among cattle in many countries. Although infected adult cows rarely show overt clinical signs, milk production may be impaired. Thus, significant production losses may occur in dairy herds with a high prevalence of fasciolosis. In this study, Bayesian hierarchical modelling was used to estimate the geospatial distribution of dairy cattle fasciolosis and its impact on milk production. The study was conducted in Galicia, the main milk producing region in Spain and a geographically heterogeneous area. The aims were: 1) to model the geospatial distribution of fasciolosis in dairy herds in the study area, 2) to identify clusters of herds with a high prevalence of fasciolosis, and 3) to assess the effect of fasciolosis on milk yield and quality. A large number of dairy cattle farms (n = 4907), of which 1660 provided production records, were surveyed. Fasciola infection status was determined by applying the MM3-SERO ELISA test to bulk tank milk samples. A high probability of infection was predicted in several zones, particularly in the centre, northeast and southeast of Galicia. Conversely, the predicted probability was very low in some parts of the northwest of the region. Infections with high within-herd prevalence (> 25% lactating cows infected) predominated. High within-herd prevalence was associated with loss of milk production (−1.387 kg/cow/ day, on average). No association between Fasciola infection and either milk fat or protein content was observed. This study has generated the first maps of the spatial distribution of the probability of Fasciola infection in dairy cattle herds in Galicia. The maps presented here can be used for reference purposes, enabling the design of better targeted fasciolosis control programmes in the region. Use of Bayesian hierarchical statistical analysis enabled us to ascertain the uncertainty of the predictions and to account for the spatial autocorrelation in the data. It also enabled us to generate maps showing the residual spatial variation in milk production, a topic that may deserve more detailed study.

Staphylococcus aureus adlb gene is associated with high prevalence of intramammary infection in dairy herds of northern Italy: A cross-sectional study

Staphylococcus aureus is a major mastitis pathogen in dairy cattle worldwide, responsible for substantial economic losses. Environmental factors, milking routine, and good maintenance of milking equipment have been described as important factors to prevent intramammary infections (IMI). Staphylococcus aureus IMI can be widespread within the farm or the infection can be limited to few animals. Several studies have reported that Staph. aureus genotypes differ in their ability to spread within a herd. In particular, Staph. aureus belonging to ribosomal spacer PCR genotype B (GTB)/clonal complex 8 (CC8) is associated with high within-herd prevalence of IMI, whereas other genotypes are generally associated with individual cow disease. The adlb gene seems to be strictly related to Staph. aureus GTB/CC8, and is a potential marker of contagiousness. We investigated Staph. aureus IMI prevalence in 60 herds in northern Italy. In the same farms, we assessed specific indicators linked to milking management (e.g., teat condition score and udder hygiene score) and additional milking risk factors for IMI spread. Ribosomal spacer-PCR and adlb-targeted PCR were performed on 262 Staph. aureus isolates, of which 77 underwent multilocus sequence typing. In most of the herds (90%), a predominant genotype was identified, especially Staph. aureus CC8 (30%). In 19 of 60 herds, the predominant circulating Staph. aureus was adlb-positive and the observed IMI prevalence was relevant. Moreover, the adlb gene was detected only in genotypes of CC8 and CC97. Statistical analysis showed a strong association between the prevalence of Staph. aureus IMI, the specific CCs, and carriage of adlb, with the predominant circulating CC and presence of the gene alone explaining the total variation. Interestingly, the difference in the odds ratio obtained in the models for CC8 and CC97 suggests that it is carriage of the adlb gene, rather than the circulation of these CCs per se, that leads to higher within-herd prevalence of Staph. aureus. In addition, the model showed that environmental and milking management factors had no or minimal effect on Staph. aureus IMI prevalence. In conclusion, the circulation of adlb-positive Staph. aureus strains within a herd has a strong effect on the prevalence of IMI. Thus, adlb can be proposed as a genetic marker of contagiousness for Staph. aureus IMI in cattle. However, further analyses using whole-genome sequencing are required to understand the role of genes other than adlb that may be involved in the mechanisms of contagiousness of Staph. aureus strains associated with high prevalence of IMI.

Alberta dairy farmers’ and veterinarians’ opinion about bovine leukemia virus control measures

A high herd and within-herd prevalence of Bovine Leukemia Virus (BLV) infections in the dairy herds of North America and the negative effects thereof caused the Alberta dairy industry to initiate the development of an on farm BLV control program. Because BLV control is dependent on the commitment of the farmer, potential barriers were identified and farmers’ and veterinarians’ points of view toward different control options were investigated to inform how the control program might be adjusted. Conversations with these stakeholders were sought and four focus groups with farmers and eleven interviews with veterinarians were conducted. Testing for BLV, the most common BLV control strategies (testing/culling/segregation/management), as well as on farm best management practices (BMP) to prevent the transmission of BLV, were discussed. The thematic analysis of these conversations resulted in the following findings: Testing of animals was considered important for BLV control, but the financial investment was prohibitive for farmers. Test and cull as well as test and segregation approaches of test positive animals were considered efficient BLV control measures, but impractical and not feasible due to the supply managed Alberta dairy industry (i.e. milk is produced based on demand), with a high prevalence. The management of test positive animals with BMP to prevent new infections and thereby decreasing the within-herd prevalence was considered the only realistic BLV control strategy. The most important barriers for suggested BMP were the cost for some BMP, the inconvenience of performing other BMP, as well as difficulties in performing some BMP consistently and well. Additionally, a lack of knowledge about BLV and its control were identified as an important barrier. On the contrary, farmers indicated being inclined to implement BMP they considered feasible or that were considered a standard within the industry. Further, if BMP increased convenience on farm, they were considered easy to implement. Farmers and veterinarians agreed in many, but not all cases. For example, the single use of examination sleeves was met with differing opinions (i.e. considered doable by farmers while veterinarians assumed it to be too costly). In conclusion, stakeholders’ awareness and communication amongst each other (e.g. veterinarians and farmers) about BLV and its control has to be highlighted in order to manage BLV infection successfully. In addition, by communicating and understanding barriers and motivators for specific BMP, important barriers could be identified (e.g. difficulties while changing needles), and solutions found (e.g. tool belt for needles), thereby improving BLV control efforts on farm.

Estimation of the value of Johne’s disease (paratuberculosis) control to Canadian dairy producers

Johne’s disease (JD), or paratuberculosis, is an infectious disorder primarily associated with cattle and sheep and resulting in significant economic losses for dairy producers. The dairy cattle herd-level prevalence in Canada has recently been estimated to be greater than 40%, but the willingness to pay for JD control practices such as testing-and-culling and vaccination among Canadian dairy producers is unknown. This study used confidential cost-of-production data from the Canadian Dairy Commission to develop a Canadian dairy production model incorporating feed, land, labor, and machinery. A second dataset from a nationally distributed questionnaire (closed in March 2020) was used to estimate individual dairy producer valuations of the reduced per-cow cost of milk production that would result from JD control. This is a novel application of compensating variation and equivalent variation (CV and EV), with dairy producers framed as consumers of production inputs and milk output as a proxy for utility. Assuming a within-herd prevalence of 12.5% and a 50% reduction of that prevalence over 10 years, it was estimated JD control has an annual value of CA$28 per cow for the average Canadian dairy producer. Within-herd prevalence, the effectiveness of control at reducing within-herd prevalence, and the time required to achieve that reduction were identified as important factors. With the same assumption of 12.5% within-herd prevalence but with 100% reductions in that prevalence, estimated values ranged from over CA$55 to over CA$90 per cow per year depending on the timeframe of the control program. When assuming a 10-year period required to achieve control, the estimated values exceeded CA$90 per cow per year in various scenarios for herds with higher within-herd prevalence (greater than 20%).

Staphylococcus aureus related to bovine mastitis in Switzerland: Clonal diversity, virulence gene profiles, and antimicrobial resistance of isolates collected throughout 2017

Staphylococcus aureus can be associated with subclinical, acute, chronic, and toxic cases of bovine intramammary infections, leading to considerable financial losses for the dairy industry in Switzerland and worldwide. In addition, milk products are one of the most common food categories implicated in staphylococcal food poisoning in humans. Detailed information on the population structure, as well as the virulence and resistance characteristics of Staph. aureus originating from bovine mastitis milk is needed to allow for source attribution and risk assessment of Staph. aureus in a food poisoning context and to improve therapeutic approaches in cattle. Our objective was to assess the population structure, phenotypic resistance patterns, and virulence and resistance gene profiles of Staph. aureus isolates from bovine mastitis milk in Switzerland. To this end, 58 Staph. aureus strains were characterized. The DNA microarray was used to test for the presence or absence of virulence and resistance genes. In addition, minimum inhibitory concentrations of various antimicrobial agents were determined by microdilution. To assess the population structure of the isolates, we determined clonal complexes (CC) using DNA microarray hybridization profiles and performed multilocus sequence typing and spa typing. The strains were assigned to 7 clonal complexes, 10 sequence types, and 11 spa types, with CC705 (43%), CC97 (33%), and CC20 (12%) representing the most common lineages and t529 (43%) and t267 (21%) representing the most common spa types. Only 1 isolate was assigned to CC8, a clonal lineage linked to high within-herd prevalence of mastitis. A total of 14% (n = 8) of strains were classified as resistant to penicillin, and 1 strain each was classified as oxacillin and pirlimycin resistant. Although no clinical breakpoints are available for the combination of kanamycin/cefalexin, growth of all strains was inhibited by the lowest combination of kanamycin/cefalexin concentrations tested (4 µg/mL of kanamycin and 0.4 µg/mL of cefalexin). One strain assigned to CC20, ST389, and t2094 exhibited resistance to penicillin, oxacillin, and pirlimycin as well as intermediate susceptibility to erythromycin and high minimum inhibitory concentration for several antimicrobial agents, for which no breakpoints were available.

Distinct phenotypic traits of Staphylococcus aureus are associated with persistent, contagious bovine intramammary infections

Staphylococcus aureus causing persistent, recurrent bovine intramammary infections are still a major challenge to dairy farming. Generally, one or a few clonal lineages are predominant in dairy herds, indicating animal-to-animal transfers and the existence of distinct pathotypic traits. The aim of this study was to determine if long term persistence and spreading of S. aureus are associated with specific phenotypic traits, including cellular invasion, cytotoxicity and biofilm formation. Mastitis isolates were collected over a 3-years period from a single dairy herd, resulting in two persistent subtypes, the high within-herd prevalent subtype ST9 (CC9)-methicillin-susceptible S. aureus (MSSA), designated HP/ST9, and the low within-herd prevalent subtype ST504 (CC705)-MSSA, designated LP/ST504. Characterization of the two different coexisting persistent subtypes showed that the following phenotypic traits are particularly associated with high within-herd prevalence: lack of capsular polysaccharide expression, high cellular invasiveness, low cytotoxicity and high biofilm/ poly-N-acetylglucosamine (PNAG) production, which may concomitantly contribute to the spreading of HP/ST9 within the herd. By contrast to HP/ST9, LP/ST504 is characterized by the formation of colony dendrites, which may help the bacteria to access deeper tissues as niches for persistence in single animals. Thus, within a single herd, two different types of persistence can be found in parallel, allowing longtime persistence of S. aureus in dairy cattle. Furthermore, this study indicates that ST9 (CC9)-MSSA strains, which are currently thought to have their primary reservoir in swine and humans, can also successfully spread to new hosts and persist in dairy herds for years.

Seasonal variation of Fasciola hepatica antibodies in dairy herds in Northern Ireland measured by bulk tank milk ELISA.

Bovine fasciolosis, caused by the infection of the trematode parasite Fasciola hepatica, remains a problem in dairy herds causing significant production losses. In this study, bulk milk tank samples were utilised to generate a comprehensive survey of the variation in liver fluke exposure over the four seasons of 2016 in Northern Ireland (NI). Samples were tested using an antibody ELISA test; within-herd prevalence levels were categorised relative to sample-to-positive ratio (S/P%). Overall, 1494 herds (~ 50% of all active dairy farms in NI) were sampled. In total, 5750 samples were tested with 91% of herds having a sample result for each season. The proportion of herds with evidence of liver fluke exposure was very high across the year, with 93.03% of all bulk milk samples having some indication of liver fluke antibody presence. A high proportion of samples (2187/5750; 38.03%) fell within the highest infection class (indicating high within-herd prevalence). There was significant seasonal variation in the mean S/P%. A multivariable random effect ordinal logit model suggested that the greatest probability of being in a higher infection class was in winter, whilst the lowest was recorded during summer. There was a significant negative association between increasing herd liver fluke infection class and herd size. Furthermore, there was significant variation in infection levels across regions of Northern Ireland, with higher infection levels in northern administrative areas. This study demonstrates the very high liver fluke exposure in this region of Europe, and that risk is not equally distributed spatially or across seasons in dairy herds.

Evaluating the impact of transmission mode, calibration level and farmer compliance in simulation models of paratuberculosis in dairy herds

Simulation models can predict the outcome of different strategies for the control and eradication of paratuberculosis (PTB) in dairy herds. Two main transmission modes have previously been used to simulate the spread of PTB: direct (contact between animals) and indirect (through the environment). In addition, previous models were calibrated to either low or high within-herd prevalence levels, which we refer to as normal and low hygiene levels, respectively. We simulated both direct and indirect transmission with the same model in both normal and low hygiene level scenarios. The effectiveness of a test-and-cull strategy was dependent on the calibration level of the simulation model, and eradication occurred less frequently with the more biologically plausible indirect transmission mode. The results were compared to within-herd prevalence records from 314 dairy herds. The prevalence in 50% of the herds varied less than 0.9% per year on average, and less than 4% in 90% of the herds. We therefore conclude that the normal-hygiene scenario best describes most dairy herds in Denmark. Finally, we simulated different levels of farmer compliance with a test-and-cull strategy and found that a 60% compliance level was not sufficient to reach eradication within 10 years.

Influence of true within-herd prevalence of small ruminant lentivirus infection in goats on agreement between serological immunoenzymatic tests

The study was conducted to evaluate influence of the true within-herd prevalence of small ruminant lentivirus (SRLV) infection on agreement beyond chance between three different types of commercial serological ELISAs. Blood samples were collected from 865 goats from 12 dairy goat herds. Serum samples were tested using three commercial ELISA kits: whole-virus indirect ELISA (wELISA), indirect ELISA based on recombined TM and CA antigens (TM/CA-ELISA), and competitive-inhibition ELISA based on SU antigen (SU-ELISA). Herds were classed into three prevalence strata of high (>50%), moderate (10–50%) and low (<10%) true within-herd prevalence of SRLV infection. The latter was estimated on the basis of results of wELISA adjusted by its sensitivity and specificity. Agreement beyond chance between the three ELISAs was assessed at two levels. First, the general agreement was determined using two coefficients corrected for chance-agreement: Cohen’s kappa and Gwet’s AC1. Then, agreement between tests was evaluated using Gwet’s AC1 separately in the three prevalence strata and compared between them by computing 95% confidence intervals for differences with a Bonferroni correction for multiple comparisons. The general agreement between the three tests was very good: wELISA and TM/CA-ELISA – Cohen’s kappa of 81.8% (CI 95%: 77.9% to 85.7%), Gwet’s AC1 of 82.7% (CI 95%: 79.0% to 86.4%); wELISA and SU-ELISA – Cohen’s kappa of 83.2% (CI 95%: 79.4% to 86.9%), Gwet’s AC1 of 83.9% (CI 95%: 80.4% to 87.5%); TM/CA-ELISA and SU-ELISA – Cohen’s kappa of 86.0% (CI 95%: 82.6% to 89.5%), Gwet’s AC1 of 86.9% (CI 95%: 83.6% to 90.1%). However, agreement between ELISAs was significantly related to the within-herd true prevalence – it was significantly lower (although still high) when within-herd true prevalence was moderate (Gwet’s AC1 between 67.2% and 78.7%), whereas remained very high, when true within-herd prevalence was either >50% (Gwet’s AC1 between 91.9% and 98.8%) or <10% (Gwet’s AC1 between 94.7% and 98.4%). Concluding, the three different commercial ELISAs for SRLV infection in goats available on the market yield highly consistent results. However, their agreement is affected by the true within-herd prevalence in a tested population, and the worse (although still high) agreement should be expected, when the percentage of infected goats is moderate.

Genomic and transcriptomic comparison between Staphylococcus aureus strains associated with high and low within herd prevalence of intra-mammary infection

BackgroundStaphylococcus aureus (Staph. aureus) is one of the major pathogens causing mastitis in dairy ruminants worldwide. The chronic nature of Staph. aureus infection enhances the contagiousness risk and diffusion in herds. In order to identify the factors involved in intra-mammary infection (IMI) and diffusion in dairy cows, we investigated the molecular characteristics of two groups of Staph. aureus strains belonging to ST8 and ST398, differing in clinical properties, through comparison of whole genome and whole transcriptome sequencing.ResultsThe two groups of strains, one originated from high IMI prevalence herds and the other from low IMI prevalence herds, present a peculiar set of genes and polymorphisms related to phenotypic features, such as bacterial invasion of mammary epithelial cells and host adaptation. Transcriptomic analysis supports the high propensity of ST8 strain to chronicity of infection and to a higher potential cytotoxicity.ConclusionsOur data are consistent with the invasiveness and host adaptation feature for the strains GTB/ST8 associated to high within-herd prevalence of mastitis. Variation in genes coding for surface exposed proteins and those associated to virulence and defence could constitute good targets for further research.

Effectiveness of a standardized footbath protocol for prevention of digital dermatitis

A footbath is the most common herd-level approach to control digital dermatitis (DD) in intensive dairy farming. However, wide variation in footbath design and protocols suggests a gap between evidence-based management practices and on-farm implementation. The objective was to evaluate effectiveness of a standardized footbath protocol in decreasing prevalence of active DD lesions in lactating dairy cattle. The protocol was based on current scientific literature, including footbath design and management. The M-stage classification system was used to score DD lesions. Scores were also combined into a simplified scoring system: no lesions (M0), active lesions (M1, M2, and M4.1), and chronic lesions (M3 and M4). A controlled intervention trial was conducted on 9 farms over 22 wk. Each farm served as its own control with data collected for 10 wk before and 12 wk after intervention. A total of 1,978 lactating cattle were assessed biweekly for DD lesions and leg cleanliness in the milking parlor. Lactating cattle were also inspected in the trimming chute at 3 time points: start, intervention, and end of trial. Intervention consisted of implementing an automated footbath that measured 3 m long, 0.50 m wide, held a fluid depth of 0.15 m, along with a weekly footbath protocol using 5% CuSO4 for 4 consecutive milkings, with footbath content replaced at a maximum of 200 cow passes. Multilevel logistic regression models for repeated measures were used to evaluate effects of the standardized footbath protocol in preventing active DD lesions. For the purpose of analysis, farms' within-herd prevalence of active DD lesions (at baseline) was assessed and categorized as low (<15%) or high (≥15%). Farms with low or high within-herd prevalence of active DD lesions at trial outset had a mean cow-level prevalence of active DD lesions of 8% (range, 2 to 13) and 31% (range, 18 to 43), respectively. At milking parlor inspections, apparent prevalence of active DD lesions decreased from the time of footbath intervention, but this effect interacted with the farms' baseline prevalence of active DD lesions. In that regard, on farms with high prevalence of active DD at baseline, apparent prevalence of active DD lesions decreased after intervention, whereas on farms with low prevalence of active DD at baseline, apparent prevalence of active DD lesions did not change. At the cow level, poor leg cleanliness was associated with higher prevalence of active DD lesions. At trimming chute inspections, prevalence of active DD lesions decreased from start to the end of the trial (22 and 14%, respectively); concurrently, prevalence of feet with no DD lesions (M0) increased (39 and 48%). We concluded that on farms with high DD prevalence, implementation of proper footbath design and improvement of footbathing management will decrease prevalence of active DD lesions and increase prevalence of feet without DD lesions. In addition, improving cow cleanliness will further result in control of active DD lesions.

Genomic characteristics of Staphylococcus aureus strains associated with high within-herd prevalence of intramammary infections in dairy cows

Staphylococcus aureus is one of the most important causes of mastitis in dairy cattle. Based on previous research, Staph. aureus genotypes with different pathogenic and contagious properties can cause intramammary infection (IMI) and coexist in the same herd. Our study aimed to compare Staph. aureus strains from herds that differed in IMI prevalence using different molecular approaches such as ribosomal spacer (RS)-PCR, multilocus sequence typing (MLST), spa typing, ribotyping, pulsed-field gel electrophoresis (PFGE), and multiplex PCR. For this purpose, 31 dairy herds with Staph. aureus IMI were selected, and 16 of these were chosen for a comparison study: the 8 high-prevalence (HP) herds had Staph. aureus IMI prevalence >28% and the 8 low-prevalence (LP) herds had an IMI prevalence <4%. A total of 650 isolates of Staph. aureus from mammary quarters of all positive cows were genotyped with RS-PCR, a technique based on amplification of a portion of the intergenic spacer 16S-23S rRNA, and a subset of 54 strains was also analyzed by multiplex PCR, ribotyping, PFGE, MLST, and spa typing. The RS-PCR analysis revealed 12 different profiles. Staphylococcus aureus strains isolated from 5 out of 8 HP herds showed a profile identical to the genotype B (GTB), described in previous studies as being strongly associated with high within-herd prevalence of Staph. aureus mastitis and the presence of the genes coding for enterotoxins sea, sed, and sej, a long x-region of spa gene, and 3 lukE fragments. Moreover, all strains isolated in the HP herds possessed genes coding for staphylococcal enterotoxins. In LP herds, a limited number of strains of 6 genotypes, different from those isolated in HP herds, were identified and GTB was not found. Within these genotypes, 4 strains were positive for the mecA gene. Preliminary results and comparison with other genotyping methods confirmed that genotyping by RS-PCR is an accurate, rapid, and inexpensive tool for future field studies on Staph. aureus mastitis strains and generates clinically relevant results.

First isolation, identification, phenotypic and genotypic characterization of Brucella abortus biovar 3 from dairy cattle in Tanzania.

BackgroundBrucellosis is a disease of worldwide public health and economic importance. Successful control is based on knowledge of epidemiology and strains present in an area. In developing countries, most investigations are based on serological assays. This study aimed at investigating a dairy herd experiencing abortions in order to establish within-herd seroprevalence to Brucella spp., identify, characterize Brucella strains by Multiple Loci Variable Number of Tandem Repeats Analysis (MLVA-VNTR) and investigate possible spillover to other species.ResultsThe within-herd seroprevalence in cattle (n = 200) was 48 % (95 % CI 41–55), using an indirect ELISA, while the Rose Bengal Test (RBT) yielded lower prevalence (21.5 %; 95 % CI 16–27). Two sheep (n = 35) and one goat (n = 50) were seropositive using ELISA while none of the dogs (n = 6) was positive with the RBT. Three Brucella were isolated from an aborted fetus and associated membranes. Real time PCR (IS711), Bruce-ladder and classical biotyping classified the isolates as B. abortus biovar 3. MLVA-VNTR revealed two different but closely related genotypes. The isolates showed unique profiles, providing the first genotypic data from Tanzania. These genotypes were not related to B. abortus biovar 3 reference strain Tulya originally isolated from a human patient in Uganda in 1958, unlike the genotypes isolated and characterized recently in Kenya. High within-herd prevalence, isolation of the pathogen and abortion confirm that B. abortus is circulating in this herd with cattle as reservoir hosts. A low seroprevalence in sheep and goats suggests a spillover of B. abortus from cattle to small ruminants in the herd.ConclusionsThis is the first isolation and characterization of B. abortus biovar 3 from a dairy cow with abortion in Tanzania. The origin of the Tanzanian genotypes remain elusive, although they seem to be related to genotypes found in Europe, Turkey and China but not related to B. abortus biovar 3 reference strain or genotypes from Kenya. Importantly, replacement heifers are commonly sourced from large farms like this to smallholder farmers, which poses risk of spread of bacteria to other herds. B. abortus is a significant zoonotic risk and animal health problem in this production system, therefore further studies on humans is recommended.

Outbreak of staphylococcal food poisoning among children and staff at a Swiss boarding school due to soft cheese made from raw milk

On October 1, 2014, children and staff members at a Swiss boarding school consumed Tomme, a soft cheese produced from raw cow milk. Within the following 7h, all 14 persons who ingested the cheese fell ill, including 10 children and 4 staff members. Symptoms included abdominal pain and violent vomiting, followed by severe diarrhea and fever. We aim to present this food poisoning outbreak and characterize the causative agent. The duration of the incubation period was dependent of the age of the patient: 2.5h in children under 10 yr of age, 3.5h in older children and teenagers, and 7h in adults. The soft cheese exhibited low levels of staphylococcal enterotoxin (SE) A (>6ng of SEA/g of cheese) and high levels of staphylococcal enterotoxin D (>200ng of SED/g of cheese). Counts of 107 cfu of coagulase-positive staphylococci per gram of cheese were detected, with 3 different Staphylococcus aureus strains being present at levels >106 cfu/g. The 3 strains were characterized using spa typing and a DNA microarray. An enterotoxin-producing strain exhibiting sea and sed was identified as the source of the outbreak. The strain was assigned to spa type tbl 3555 and clonal complex 8, and it exhibited genetic criteria consistent with the characteristics of a genotype B strain. This genotype comprises bovine Staph. aureus strains exclusively associated with very high within-herd prevalence of mastitis and has been described as a major contaminant in Swiss raw milk cheese. It is therefore highly likely that the raw milk used for Tomme production was heavily contaminated with Staph. aureus and that levels further increased due to growth of the organism and physical concentration effects during the cheese-making process. Only a few staphylococcal food poisoning outbreaks involving raw milk products have been described. Still, in view of this outbreak and the possible occurrence of other foodborne pathogens in bovine milk, consumption of raw milk and soft cheese produced from raw milk constitutes a health risk, particularly when young children or other members of sensitive populations are involved.

A longitudinal study investigating the prevalence of Staphylococcus aureus genotype B in seasonally communal dairy herds

Staphylococcus aureus is a major mastitis-causing pathogen. Various genotypes have been recently identified in Switzerland but Staph. aureus genotype B (GTB) was the only genotype associated with high within-herd prevalence. The risk of introducing this Staph. aureus genotype into a herd may be increased by frequent animal movements. This may also be the case when cows from different herds of origin are commingled and share their milking equipment for a limited period of time. The aim of the present study was to determine the prevalence of Staph. aureus GTB in seasonally communal dairy herds before and after a summer period when dairy farming is characterized by mixing cows from different herds of origin in 1 communal operation. In addition, the environment was investigated to identify potential Staph. aureus GTB reservoirs relevant for transmission of the disease. A total of 829 cows from 110 herds of origin in 9 communal operations were included in the study. Composite milk samples were collected from all cows during the first or second milking after arrival at the communal operation and again shortly before the end of the season. Swab samples from the environment, involved personnel, and herding dogs present were collected before the cows arrived. At the end of the season, sampling of personnel was repeated. All samples were analyzed for the presence of Staph. aureus GTB using an established quantitative PCR. At the beginning of the season, Staph. aureus GTB-positive cows were identified in 7 out of 9 communal operations and the within-communal operation prevalence ranged from 2.2 to 38.9%. At the second sampling, all communal operations were Staph. aureus GTB positive, showing within-communal operation prevalence from 1 to 72.1%. The between-herd of origin prevalence increased from 27.3 to 56.6% and the cow-level prevalence increased from 11.2% at the beginning of the season to 29.6% at the end of the season. On 3 different communal operations, Staph. aureus GTB-positive swabs from seasonally employed personnel were identified at the end of the season. The results indicate that Staph. aureus GTB can easily spread in communal operations when cows from different herds of origin are mixed during the summer season. Effective management measures need to be designed to prevent the spread of Staph. aureus GTB in seasonally communal herds.

Schmallenberg virus in Dutch dairy herds: Potential risk factors for high within-herd seroprevalence and malformations in calves, and its impact on productivity

In November 2011, the new orthobunyavirus Schmallenberg virus (SBV) was identified in dairy cows that had induced fever, drop in milk production and diarrhoea in the Netherlands (Muskens et al., 2012. Tijdschrift voor Diergeneeskunde 137, 112–115) and a drop in milk production in cows in Northwestern Germany (Hoffmann et al., 2012. Emerging Infectious Diseases 18 (3), 469–472), in August/September 2011. This study aimed at quantifying risk factors for high within-herd prevalence of SBV and SBV-induced malformations in newborn calves in dairy herds in the Netherlands. Additionally, the within-herd impact of SBV infection on mortality rates and milk production was estimated.A case-control design was used, including 75 clinically affected case herds and 74 control herds. Control herds were selected based on absence of malformations in newborn calves and anomalies in reproductive performance. SBV-specific within-herd seroprevalences were estimated. Risk factors for high within-herd SBV seroprevalence (>50%) and the probability of malformed newborn calves in a herd were quantified. In addition, within-herd impact of SBV with regard to milk production and mortality was estimated.Animal-level seroprevalence was 84.4% (95% confidence interval (CI): 70.8–92.3) in case herds and 75.8% (95% CI: 67.5–82.5) in control herds. Control herds that were completely free from SBV were not present in the study. Herds that were grazed in 2011 had an increased odds (OR 9.9; 95% CI: 2.4–41.2)) of a high seroprevalence (>50%) compared to herds that were kept indoors. Also, when grazing was applied in 2011, the odds of malformations in newborn calves tended to be 2.6 times higher compared to herds in which cattle were kept indoors. Incidence of malformations in newborn calves at herd level was associated with both within-herd seroprevalence and clinical expression of the disease in adult cattle.The rate of vertical transmission of SBV to the fetus once a dam gets infected seemed low. A total of 146 stillborn or malformed calves were submitted by 65 farmers during the study period, of which 19 were diagnosed as SBV-positive based on pathological investigation and/or RT-qPCR testing of brain tissue. Based on these results combined with calving data from these herds we roughly estimated that at least 0.5% of the calves born between February and September 2012 have been infected by SBV.A drop in milk production was observed between the end of August 2011 and the first half of September (week 35–36), indicating the acute phase of the epidemic. During a 4-week period in which SBV infection was expected to have occurred, the total loss in milk production in affected dairy herds was around 30–51kg per cow. SBV had no or limited impact on mortality rates which was as expected given the relatively mild expression of SBV in adult cows and the low incidence of malformations in newborn calves.

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.

Seroepidemiology of bovine herpesvirus 1 (BHV1) infection among Estonian dairy herds and risk factors for the spread within herds

The objectives of this study were to reassess the herd level and within-herd prevalence of bovine herpesvirus 1 (BHV1) infection in Estonian dairy cattle, estimate the sensitivity and specificity of the enzyme-linked immunosorbent assay (ELISA) for bulk tank milk (BTM) testing and determine the risk factors related to high prevalence of the infection in herds. To estimate the herd prevalence, BTM samples from each of the 1,205 herds that sell milk to dairy companies were analysed for BHV1 antibodies. One hundred and three herds with known BHV1 infection status were selected to estimate within-herd prevalence and to calculate the sensitivity and specificity of BTM ELISA. In these herds serum samples were collected from cows and youngstock, together with BTM samples. A commercial blocking ELISA test was used to analyse samples for antibodies against BHV1. A questionnaire was completed to collect herd data. The sensitivity and specificity of the BTM ELISA were 76.5% and 97.2%, respectively, and the true herd prevalence of BHV1 was calculated to be 22.0%. The herd prevalence increased significantly with herd size, being 3.4% in the smallest category (less than 20 cows) and 85.7% in herds of size over 400. The mean within-herd prevalence was 37.8% (range 1–100, median 31.5). The mean within-herd prevalence increased with herd size. Data from 59 infected herds was used to determine the risk factors associated with high within-herd prevalence (>50%) of BHV1, using logistic regression analysis. As, in some infected herds, the youngstock were uninfected, risk factors for the presence of BHV1 among youngstock from 6 months until calving were analysed. The results indicate the importance of iatrogenic spread of the virus, since the overall within-herd prevalence was higher in those herds in which a veterinarian was an employee of the farm and an inseminator worked only for the particular farm. The presence of bovine viral diarrhoea virus (BVDV) in a herd was associated with a higher prevalence of BHV1.