Negative Herds Research Articles

An epidemiological investigation into the reasons for high bovine tuberculosis incidence in cattle herds of the Burren, Ireland, prior to 2020

Herd-level bovine tuberculosis (bTB) incidence was examined in the Burren, an area in the west of Ireland where herd owners practice distinctive transhumance practices, with upland winter grazing. Prior to the initiation of our study in 2020, bTB incidence had for many years been unusually high in the Burren in comparison with the rest of the country, although the most recent figures have come down to being closer to the national average. Using data from the period prior to 2020, we mapped bTB infection in Burren herds alongside a range of indicators thought to have an association with it - herd size, herd density, herd type, cattle movement, and badger (Meles meles) population and control data, as well as rainfall and altitude. We also looked at how summary statistics for these variables differed when Burren herds with a history of bTB were compared to other Burren herds, as well as bTB positive and negative herds from outside the Burren. We found that for many indicators Burren herds would be expected to be low risk when compared to other herds in Ireland. An exception to this was for rainfall: hot spot areas for bTB in the Burren were found in areas of higher rainfall, on average herds in the Burren experienced more rainfall than those outside it, and bTB herds in the Burren experienced higher rainfall than non-bTB herds. Separately, for Burren herds only, a logistic regression model was developed to explain bTB breakdown occurrence using a matched case-control approach. Cases were herds which had experienced a new bTB breakdown between 2015 and 2019 (n = 260) and these were matched on herd type and herd size with the same number of herds not experiencing a breakdown during this period. This showed that, of a range of exogenous variables, rainfall was the most strongly associated with herd-level bTB incidence. These results suggest that high levels of exposure to inclement weather, and/or better environmental survival of Mycobacterium bovis in the environment, may contribute to high bTB rates in the Burren. However, as rainfall showed a highly aggregated distribution, this relationship may be due to an unmeasured factor correlated with it. Mapping and graphical output suggested that, although herd sizes in the Burren were on average lower than nationally, within the Burren they were higher in areas of higher prevalence, suggesting that mechanisms associated with herd size, such as increased contacts between and within herd, and with wildlife, may also play a role.

Assessing the feasibility of test-and-cull and test-and-segregation approaches for the control of high-prevalence bovine tuberculosis in Ethiopian intensive dairy farms

Bovine tuberculosis (bTB) is endemic and has a substantial impact on the livestock sector in Ethiopia and other low and middle-income countries (LMICs). With a national emphasis on dairy farm intensification to boost milk production and spur economic growth, the incidence of bTB is anticipated to rise. However, Ethiopia, like other LMICs, lacks a comprehensive national bTB control strategy due to the economic and social infeasibility of traditional test-and-cull (TC) approaches. To inform the development of such a strategy, we evaluated the effectiveness and feasibility of TC and test-and-segregation (TSg) strategies for bTB control on Ethiopian dairy farms. A TC approach was used at Farm A [N = 62; comparative cervical test (CCT) > 4 mm, starting prevalence 11.3%] while TSg was implemented at Farm B (N = 45; CCT > 4 mm, prevalence 22.2%), with testing intervals of 2–4 months. Both strategies achieved a reduction in bTB prevalence to 0%, requiring seven rounds of TC over 18 months at Farm A, and five rounds of TSg over 12 months at Farm B’s negative herd. The results show that adopting more sensitive thresholds [CCT > 0 mm or single cervical test (SCT) > 2 mm] during later rounds was pivotal in identifying and managing previously undetected infections, emphasizing the critical need for optimized diagnostic thresholds. Cost analysis revealed that TC was approximately twice as expensive as TSg, primarily due to testing, labor, and cow losses in TC, versus construction of new facilities and additional labor for TSg. This underscores the economic and logistical challenges of bTB management in resource-limited settings. Taken together, our study highlights an urgent need for the exploration of alternative approaches including TSg and or vaccination to mitigate within herd transmission and enable implementation of bTB control in regions where TC is not feasible.

District-wide herd sanitation and eradication of intramammary Staphylococcus aureus genotype B infection in dairy herds in Ticino, Switzerland

The present study demonstrates successful herd sanitation and eradication of contagious mastitis caused by Staphylococcus aureus genotype B (S. aureus GTB) in an entire Swiss district (Ticino) including 3,364 dairy cows from 168 farms. Herd sanitation included testing of all cows using a highly GTB specific and sensitive qPCR assay, implementation of related on-farm measures, appropriate antibiotic therapy of GTB-positive cows and culling of therapy-resistant animals, respectively. A treatment index was used as an objective criterion to select GTB-positive cows eligible for culling and replacement payment. 62 herds (37%) were initially GTB-positive with a cow prevalence between 10% and 100% and were submitted to sanitation. Twenty mo after the start of the campaign, all these herds were free from S. aureus GTB, whereby 73% of them were sanitized during the first 7 mo. At the cow level, a total of 343 animals were infected. 50 of them were immediately culled and financially compensated based on their treatment index value. The remaining 293 cows were intramammarily treated with antibiotics either during lactation using the combination of cephalexin-kanamycin or penicillin-gentamicin or at dry-off using cloxacillin. Out of these cows, 275 (93.9%) were treated successfully meaning that their milk was twice GTB-negative by qPCR after therapy. For lactational treatment, control samples were taken ≥10 and ≥20 d after treatment, for dry off treatment ≥14 and ≥24 d after parturition. Neither lactation number nor SCC before treatment of the cow nor the type of therapy were associated with therapeutic cure. Using data of 30 GTB-positive and 71 GTB-negative herds (1855 observations), the impact of GTB sanitation on bulk tank milk SCC (BTSCC) was evaluated applying a linear mixed statistical model. In the year before sanitation, BTSCC was always higher in GTB positive than in GTB negative herds. After the start of the campaign, BTSCC declined rapidly in the herds under GTB sanitation and achieved values that no longer differed statistically from those of GTB-free herds after only 2 mo, remaining very similar for the rest of the campaign. The farmers were very satisfied with the outcome of the campaign as all GTB positive herds could be sanitized rapidly, sanitation was sustainable, and milk quality increased.

Estimation of diagnostic sensitivity and specificity of abattoir registrations and bulk tank milk ELISA as herd-level tests for Fasciola hepatica using Bayesian latent class modelling

The common liver fluke, Fasciola hepatica, is a trematode parasite found worldwide, typically with a focal distribution due to its requirement for suitable climatic and environmental conditions to complete its lifecycle. Bovine fasciolosis causes suboptimal production and economic losses, including liver condemnation at slaughter. The lack of reliable diagnostic methods is a disadvantage to the increasing demand for surveillance and control. The aim of this study was to evaluate the diagnostic accuracy of bulk tank milk (BTM) antibody testing and aggregated abattoir registrations (AAR) of liver fluke as herd-level tests for F. hepatica infection using Bayesian latent class models. Data from the abattoirs in 2019–2021 and BTM, sampled in the winter of 2020/2021, were collected from 437 herds on the southwest coast of Norway. The BTM samples were analysed with the SVANOVIR® F. hepatica-Ab ELISA test, with results given as an optical density ratio (ODR), and later dichotomized using the recommended cut-off value from the test manufacturer (ODR ≥0.3). Based on the BTM ELISA test, 47.8% of the herds tested positive. The AAR test was defined as the herd-level proportion of female slaughtered animals registered with liver fluke infection during the study period. For this test, three cut-offs were used (a proportion of 0.05, 0.1 and 0.2). The herds were split into two subpopulations (“Coastal” and “Inland”), which were expected to differ in true prevalence of F. hepatica infection based on climate-related and geographical factors. The diagnostic accuracies of both tests were estimated using Bayesian latent class models with minimally informative priors. Post-hoc analysis revealed that the maximum sum of sensitivity (Se) and specificity (Sp) of the tests was achieved with a herd-level proportion of ≥0.1 registered with liver fluke as the AAR test. Using this cut-off, the median estimate for the diagnostic accuracy of the BTM ELISA was 90.4% (84.0–96.2 95% Posterior Credible Interval (PCI)) for Se and 95.3% (90.6–100% PCI) for Sp, while the median estimate of Se for AAR was 87.5% (81.4–93.1% PCI) and the median estimate of Sp for AAR was 91.0% (85.2–96.5% PCI). The cut-off evaluation of the SVANOVIR® F. hepatica-Ab ELISA test for BTM confirmed the manufacturer's recommended cut-off of ODR ≥0.3 to denote positive and negative herds. This study suggests that AAR and BTM ELISA test can be used as herd-level tools to monitor liver fluke infection, so that appropriate interventions against infection can be implemented as necessary.

Práticas de biosseguridade associadas com infecção do vírus da diarreia viral bovina em rebanhos leiteiros no Brasil

ABSTRACT: This study determined the association between biosecurity practices and the status of bovine viral diarrhea virus (BVDV) in dairy production systems. Approximately 280 herds were screened for BVDV virus detection. Following the screening, 68 herds were selected to identify individual BVDV PI animals using an ear notch biopsy and ELISA-antigen. All offspring of the last generation were tested, and the maternal lineage of positive cases was examined. A questionnaire on BVDV biological risk assessment was completed. A multiple correspondence analysis (MCA) was conducted to determine the association between herds with or without BVDV circulation and biosecurity practices. The MCA revealed that farms with virus circulation lacked knowledge about the disease and wrongly perceived their herds as protected, while farms without virus circulation were aware of the disease but considered their herds unprotected. Vaccination practices differed between positive and negative herds, with positive herds using vaccines only for reproductive diseases and negative herds vaccinating for respiratory and reproductive issues. Biosecurity practices such as frequent visitation, contact between animals of different ages, and annual introduction of new animals were linked to viral circulation, while virus-free herds implemented measures like controlled visitation, no contact between different age groups, and quarantine. Lastly, herds with virus circulation acquired pregnant females without prior testing. This study emphasized the crucial role of biosecurity practices in controlling BVDV in dairy herds. It highlighted the sharper risk perception and better application of biosecurity practices in negative herds compared to BVDV-positive herds.

Bovine trichomoniasis overview and testing

Bovine trichomoniasis, caused by Tritrichomonas foetus, is a venereal disease of cattle, resulting in infertility, pregnancy losses, abortion and delayed calving seasons. The disease has global distribution, with greater prevalence in areas in which natural service is the primary means of reproduction. When ar­tificial insemination, with appropriate hygiene practices, is im­plemented, as well as aggressive testing and culling of positive bulls, control and eradication of the disease is possible. Addi­tionally, minimizing co-mingling of animals with unknown sta­tus through herd additions or on public grazing lands will help decrease the likelihood of introduction into negative herds, and contribute significantly to the decrease in prevalence in the local region. Testing methods have improved over the last 100 years; however, proper sample collection and handling are still paramount to gaining true positive results in infected bulls.

Bovine paratuberculosis: results of a control plan in 64 dairy farms in a 4-year period

Paratuberculosis is considered one of the most economically devastating infectious diseases of domestic livestock, and the most effective control strategy is a combination of ‘test-and-cull’ and on-farm biosecurity measures. In Italy, a Voluntary National Control Plan (VNCP) and guidelines have been introduced to reduce the impact of the disease, and farmers can voluntarily enroll in the control plan. The main aims of this study were: i) the description of the trend over a 4-year period on total, within-herd (WH) and between herd (BH) apparent seroprevalences observed in 64 dairy herds members of a mutual company located in Italy after the introduction of a proposed “Customized Control Plan” (CCP); ii) the evaluation of its effectiveness in terms of percentage of participating farms that decided to join the VNCP. Analyses on serum samples were performed with Enzyme-Linked Immuno Sorbent Assay (ELISA) method and revealed a general decrease in both total, WH and BH apparent seroprevalence. Total average apparent seroprevalence decreased from 2.39% in 2017 to 1% in 2020. Negative herds raised from 51.9% in 2017 to 71.1% in 2020, while farms with WH apparent seroprevalence > 5% decreased from 17.3% in 2017 to 4.4% in 2020. BH apparent seroprevalence decreased from 51.2% in 2017 to 29.2% in 2020. Among the 52 out of 64 herds that accepted to continue the proposed CCP after the first year, 41 (78.8%) joined in 2020 the VNCP, that assessed the health ranking of the herds. The results provide evidence that a control plan based on a farm-specific strategy and a subsidized testing process can effectively reduce the impact of paratuberculosis in dairy herds, especially in convincing farmers to continue in paratuberculosis control by joining the VNCP, including them in a National context and increasing their awareness of the disease.

Coxiella burnetii in dairy cattle herd: farming practices and infection dynamics

Abstract Q fever is a widespread disease affecting reproduction in dairy cattle. Several risk factors can increase the possibility of the herd becoming infected and the persistence of infection. The aim of this study was to characterise the dynamics of C. burnetii infection in affected herds and to evaluate reproduction problems and risk factors using a questionnaire within infected and infection-free dairy herds. In total, 25 farms that were serologically positive or shedding C. burnetii DNA in milk and 14 C. burnetii infection-free farms in Latvia answered the questionnaire. Ten positive herds were studied by testing individual blood and milk samples from up to 10 animals at two times separated by 7.5 to 13 months. The number of serologically positive and suspicious animals was higher in the second sampling even though several animals were culled. In the positive herds, the percentage of dystocia, stillborn calves and abortions during the last year was significantly (p=0.001; 0.01; 0.005, respectively) higher than in the negative herds. Several significant factors were found for the presence of Q fever infection, such as the herd size, the regional population density of ruminants, artificial ventilation systems, and frequent farm visitors. Deratisation was less practised in the negative farms and the presence of ticks was observed more often in the negative farms. Only two identified risk factors can be mitigated – the ventilation system and frequent farm visitors. The other factors cannot be changed arbitrarily but they have to be taken into account by decision makers.

A case–control study regarding factors associated with digital dermatitis in Norwegian dairy herds

BackgroundDigital dermatitis (DD) is a contagious bovine foot disease causing painful lesions, lameness, and reduced animal welfare. Previous studies indicate a complex aetiology of the disease. The aim of this study was to compare DD negative and DD positive herds to identify factors associated with DD in Norwegian dairy herds by analysing data obtained in a questionnaire and data recorded in the Norwegian Dairy Herd Recording System (NDHRS). The questionnaire was e-mailed to the owners of all 380 herds recorded with DD in 2019 and to 1530 randomly selected herds with no recordings of DD. Altogether 559 dairy herds were included in the final study population, of which 113 was classified as DD positive (≥ one cow recorded with DD in NDHRS) and 446 as DD negative. When stratified by housing system, the ratio between DD positive and negative herds was 108/281 in free stalls and 5/165 in tie stalls. Multilevel logistic regression was used to model the association between potential risk factors and variables related to the detection and diagnosis of DD, and the outcome in the free-stall population. Geographical area (county) was included in the model as a random effect.ResultsIn the final study population 108/113 (96%) of the DD positive herds were housed in free stalls versus 5/113 (4%) in tie stalls. The free-stall herds’ DD status was associated with purchase of cattle during the last 5 years (baseline: 0 animals, OR = 2.30 for category 12–27 animals, OR = 4.34 for 28–52 animals, and OR = 5.39 for ≥ 53 animals). The DD status was also associated with claw trimming frequency (Baseline: 1 < 2/year, OR = 0.41 for category < 1/year, and OR = 4.09 for ≥ 3/year), whether the claw trimming was done by a certified professional trimmer or not (baseline: ≤ 90% of the cows, OR = 3.98 for category ≥ 90% of the cows), cleaning of feet in the chute before trimming (baseline: no cleaning, OR = 1.98 for category cleaning), and alley flooring (baseline: slatted floor, OR = 2.36 for category solid floor).ConclusionsDigital dermatitis was far more frequent in Norwegian dairy herds housed in free stalls versus those housed in tie stalls. In the free-stall herds purchase of cattle, increasing trimming frequency, use of certified professional trimmer, cleaning of the feet in the chute, and solid flooring in the alleys were associated with increased odds of recorded DD.

Environmental detection of Mycoplasma hyopneumoniae in breed-to-wean farms

There is a need to develop cost-effective and non-invasive approaches to sample large populations to evaluate the disease status of breeding herds. In this study we assessed the detection of the M. hyopneumoniae genetic material in environmental surfaces and air of farrowing rooms, and skin (udder, snout and vagina) of lactating sows at weaning, in farms having different M. hyopneumoniae infection status (negative, positive sub-clinically infected and positive clinically affected). Mycoplasma hyopneumoniae was detected in air, air deposition particles, dam and stall surfaces of the positive clinically affected herd. Mycoplasma hyopneumoniae could only be detected in dam and stall surfaces in sub-clinically infected herds. Mycoplasma hyopneumoniae was not detected in all samples collected in the negative herd. The cycle threshold of the positive PCR samples were not statistically different between sample types or farms. However, a significant difference (p < 0.05) was observed in the percentage of positive samples between the positive clinically affected farm and the rest. Likewise, M. hyopneumoniae was detected in the environment and surfaces at weaning in positive breeding herds. Further testing and validation is recommended for environmental and surface samples before they can be employed as part of the M. hyopneumoniae diagnostic process. In addition, results from this study highlight potential sources of M. hyopneumoniae infection for piglets in breeding herds, especially during an outbreak.

Impact of co-evolution of negative vaccine-related information, vaccination behavior and epidemic spreading in multilayer networks

In this paper, we establish a three-layer coupled network model to analyze the co-evolution of negative vaccine-related information, vaccination behavior, and epidemic spread. The three layers are used to represent negative vaccine-related information dissemination, vaccination behavior diffusion and epidemic propagation, respectively. The comprehensive impact of vaccination costs and herd mentality are firstly considered in the diffusion of vaccination behavior. Then, we use the micro-Markov chain (MMC) method to derive the system dynamics equations and the epidemic threshold. The analytical results show that the proportion of efficaciously vaccinated individuals and the topology of the epidemic spread layer both determine the epidemic threshold. Finally, a large number of simulation experiments are conducted to investigate the impact of various factors on the epidemic spread, which include negative vaccine-related information diffusion, rational judgment and herd mentality during the process of vaccination decision, and cost of vaccine. The results demonstrate the role of negative vaccine-related information diffusion and vaccination behavior spread in epidemic control, and may provide some valuable clues for policymakers to formulate appropriate measures to prevent and control epidemics.

Spatial and environmental analysis of Mycobacterium avium infected dairies in Antioquía (Colombia)

The present study aimed to describe the spatial distribution of Mycobacterium avium subsp. paratuberculosis (MAP) in dairy herds, and to detail environmental variables taken as refer- ence of the physical background of the study area, specifically those related to MAP-qPCR positive herds located in six municipalities of the northern region of the Province of Antioquia (Colombia), based on environmental sampling and qPCR analysis. The study herds (n = 386) were located in 63 different districts from six municipalities. Participant herds were visited once between June and October (2016) to collect an environmental sample, and iden- tification of MAP was achieved using a duplex quantitative real-time PCR method. Rainfall trends, day and nighttime surface temperature, and vegetation cover index were taken as environmental references of the physical background of the study area. In addition, distribution maps of MAP-qPCR positive and negative herds, as well as maps of temperature variations and vegetation cover, were constructed. As a result, there was an increase in MAP-positive herds in the North-West, South, and Southeast of the study area. An overall high rainfall regime was found and day- and nighttime surface temperatures showed important variations during sampling months. No evidence of management of the vegetation cover was found, in both pastures and areas with native vegetation, except for a conservancy area. In conclusion, the general environmental conditions, where the detection of MAP-positive herds is most likely to happen, were reported herein, considering approaches using the same (or a very ap- proximate) sample collection and handling, and molecular detection method.

Evaluation of risk-based surveillance strategies for Salmonella Dublin in Danish dairy herds by modelling temporal test performance and herd status classification errors

The potential risk-based improvement of the Salmonella Dublin surveillance programme in Danish dairy herds was investigated, considering herd status misclassifications due to testing errors. The programme started in October 2002. Currently (early 2021) all dairy herds are classified based on quarterly bulk tank milk (BTM) testing with an indirect antibody ELISA (iELISA). Over the last two decades, the prevalence of herds classified as “likely infected” (levels 2,3) reduced remarkably. However, since 2015, the apparent prevalence has increased again, calling for improved surveillance and control to protect animal and human health. A deterministic simulation model based on data (2018–2019) from 2283 dairy herds in level 1 (“most likely free from infection”), was developed to estimate status misclassifications as false negative (FN) and false positive (FP) herds, under two testing strategies. These were: (A) the current system based on quarterly BTM testing only, and (B) an alternative strategy based on additional blood testing of up to eight calves, within herds at high risk of infection (HR). Both strategies were evaluated using three risk classification methods (I to III) and four sensitivity analysis scenarios (SA1-4), where different temporal performances were simulated for the iELISA in BTM. To apply strategy B, the best high-risk classification method (II), which combined managerial applicability and minimized errors, would require testing approximately 1000 calves across 127 HR herds. In that case, strategy A would cause 3 FNs and 67 FPs, by assuming annual BTM sensitivity (BTMSe) 95% conditional on a 1-year disease history and specificity (BTMSp) 97%. Whereas strategy B could cause a similar number of FNs, but 7 FPs more, assuming a sensitivity (Se) of 77% and specificity (Sp) of 99% in individual blood-samples (SA1). Assuming also quarterly BTMSe 53% and BTMSp 99.9% (SA4), strategy A derived 28 FNs and 2 FPs, while strategy B resulted in 6 FNs less and 8 FPs more. Therefore, strategy B could improve early detection of infected HR herds, while strategy A would avoid more unnecessary restrictions in false-positive herds. This improves knowledge on the potential use of additional blood testing in HR herds and illustrates how deterministic modelling can be used to improve disease surveillance and control.

890The prevalence and risk factors for Coxiella burnetii on commercial dairy goat farms in Australia

Abstract Background Despite the potentially important role that intensively managed dairy goats play in the spread of Q fever, the prevalence of Coxiella burnetii among dairy goat herds in Australia is largely unknown. The aim of this cross-sectional study was to estimate the prevalence of coxiellosis-positive dairy goat herds in Australia and to identify risk factors associated with coxiellosis positivity. Methods Owners or managers of commercial dairy goat herds were contacted and asked to complete a questionnaire about risk factors for coxiellosis and to provide a bulk tank milk (BTM) sample. BTM samples were tested using an enzyme-linked immunosorbent assay (ELISA) and real-time quantitative polymerase chain reaction (RT-PCR) targeting the Com1 and IS1111 sections of the C. burnetii genome. Questionnaire responses from coxiellosis positive and coxiellosis negative herds were compared using frequency cross-tabulations and multivariable logistic regression. Results Herd managers from 49 of the 61 commercial dairy goat herds in Australia took part in the study. Of this group, three BTM samples were found to be both ELISA and RT-PCR positive. Two BTM samples were ELISA positive but RT-PCR negative. There were 10 (95% CI 4.4 to 22) C. burnetii positive herds per 100 herds at risk. Conclusions The prevalence of coxiellosis among commercial dairy goat farms in Australia is relatively low. Key messages The Australian dairy goat industry should focus on biosecurity measures and risk management plans to reduce the probability of C. burnetii introduction.

Evaluation of the Efficacy of BCG in Protecting Against Contact Challenge With Bovine Tuberculosis in Holstein-Friesian and Zebu Crossbred Calves in Ethiopia.

Bovine tuberculosis (bTB) is prevalent in intensive dairy farms in Ethiopia. Vaccination could be an alternative control approach given the socio-economic challenges of a test-and-slaughter control strategy. The efficacy of the BCG was evaluated on 40 Holstein-Friesian (HF) and zebu crossbred calves recruited from single intradermal cervical comparative tuberculin (SICCT) test negative herds and randomly allocated into two groups. Twenty-two calves were vaccinated within 2 weeks of age, and 18 were kept as a control. Six weeks post-vaccination, the two groups were exposed and kept mixed with known SICCT test positive cows for 1 year. Immune responses were monitored by interferon gamma (IFN-γ) release assay (IGRA), SICCT test, and antibody assay. Vaccinated calves developed strong responses to the SICCT test at the sixth week post-vaccination, but did not respond to ESAT-6/CFP-10 peptide antigen-based IGRA. During the exposure, IFN-γ response to the specific peptide cocktail [F(2.44, 92.67) = 26.96; p < 0.001] and skin reaction to the specific proteins cocktail [F(1.7, 64.3); p < 0.001] increased progressively in both groups while their antibody responses were low. The prevalence of bTB was 88.9% (95% CI: 65.3–98.6) and 63.6% (95% CI: 40.7–83.8) in the control and vaccinated calves, respectively, based on Mycobacterium bovis isolation, giving a direct protective efficacy estimate of 28.4% (95% CI: −2.7 to 50.1). The proportion of vaccinated calves with lesion was 7.0% (34/484) against 11.4% (45/396) in control calves, representing a 38% (95% CI: 5.8–59.4) reduction of lesion prevalence. Besides, the severity of pathology was significantly lower (Mann–Whitney U-test, p < 0.05) in vaccinated (median score = 2.0, IQR = 0–4.75) than in control (median score = 5, IQR = 3.0–6.25) calves. Moreover, survival from M. bovis infection in vaccinated calves was significantly (log-rank test: χ2 = 6.749, p < 0.01) higher than that of the control calves. In conclusion, the efficacy of BCG was low, but the reduced frequency and severity of lesion in vaccinated calves could suggest its potential role in containing onward transmission.

Successful Eradication of Porcine Epidemic Diarrhea in an Enzootically Infected Farm: A Two-Year Follow-Up Study.

Porcine epidemic diarrhea virus (PEDV) has negatively affected the welfare of animals and their productivity in South Korea for three decades. A shortage of effective control measures has led to the virus becoming endemic in domestic pig populations. This study aimed to describe how our intervention measures were implemented for PEDV elimination in an enzootically infected farm. We operated a risk assessment model of PEDV recurrence to obtain information about the virus itself, herd immunity, virus circulation, and biosecurity at the farm. Next, we conducted a four-pillar-based two-track strategy to heighten sow immunity and eradicate the virus, with longitudinal monitoring of immunity and virus circulation, involving strict biosecurity, prime-boost pre-farrow L/K/K immunization, all-in-all-out and disinfection practices in farrowing houses, and disinfection and gilt management in wean-to-finish barns. In particular, we observed a high prevalence and long-term survival of PEDV in slurries, posing a critical challenge to PED eradication and highlighting the necessity for consecutive testing of barn slurry samples and for the management of infected manure to control PEDV. Genetic analysis of PEDVs in this farm indicated that genetic drift continued in the spike gene, with a substitution rate of 1.683 × 10−4 substitutions/site/year. Our study underlines the need for active monitoring and surveillance of PEDV in herds and their environments, along with the coordinated means, to eliminate the virus and maintain a negative herd. The tools described in this study will serve as a framework for regional and national PED eradication programs.

The Irish Programme to Eradicate Bovine Viral Diarrhoea Virus-Organization, Challenges, and Progress.

A mandatory national Irish bovine viral diarrhoea (BVD) eradication programme, coordinated by Animal Health Ireland, commenced in 2013. Key decisions and programme review are undertaken by a cross-industry Implementation Group (BVDIG) supported by a Technical Working Group. Ear notch tissue is collected from all new-born calves using modified official identity tags, supplemented by additional blood sampling, including for confirmatory testing of calves with initial positive results and testing of their dams. Testing is delivered by private laboratories in conjunction with the National Reference Laboratory, with all results reported to a central database. This database manages key elements of the programme, issuing results to herdowners by short message service messaging supplemented by letters; assigning and exchanging animal-level statuses with government databases of the Department of Agriculture, Food and the Marine to enable legislated restrictions on animal movements; assigning negative herd status based on test results; generating regular reports for programme management and evaluation and providing herd-specific dashboards for a range of users. Legislation supporting the programme has been in place throughout but has not thus far mandated the slaughter of persistently infected (PI) calves. A key challenge in the early years, highlighted by modeling, was the retention of PI animals by some herd owners. This has largely been resolved by measures including graduated financial supports to encourage their early removal, herd-level movement restrictions, ongoing programme communications and the input of private veterinary practitioners (PVPs). A framework for funded investigations by PVPs in positive herds was developed to identify plausible sources of infection, to resolve the status of all animals in the herd and to agree up to three measures to prevent re-introduction of the virus. The prevalence of PI calves in 2013 was 0.66%, within 11.3% of herds, reducing in each subsequent year, to 0.03 and 0.55%, respectively, at the end of 2020. Recent regulatory changes within the European Union for the first time make provision for official approval of national eradication programmes, or recognition of BVD freedom, and planning is underway to seek approval and, in due course, recognition of freedom within this framework by 2023.

Managing BVDV at the herd level part 2

Bovine viral diarrhoea virus (BVDV) is a costly disease and its eradication is the focus of the English voluntary scheme, BVDFree. The process of identifying a farm as negative or not negative is broken down into four stages using ADAM: assess disease risk; define herd status; action plan for BVD control; monitor progress. This process is based on the long-established protocols defined by Cattle Health Certification Standards. The latter two stages are discussed in this article with the first two discussed in part 1 of this series. Initial testing will identify the farm as negative or not negative for BVDV. Herds that are not negative will require further testing, which will often take the form of screening for a persistently infected (PI) animal, or ‘PI hunt’, by testing every animal in the herd either directly or indirectly for the presence of virus. Maintaining and monitoring BVDV status will require ongoing testing, tag and test, or check test at the farmer's choice, and continued attention to biosecurity, especially in negative herds that are not vaccinated.

Survey of Candidate Single-Nucleotide Polymorphisms in SLC11A1, TLR4, NOD2, PGLYRP1, and IFNγ in Ankole Longhorn Cattle in Central Region of Uganda to Determine Their Role in Mycobacterium avium Subspecies paratuberculosis Infection Outcome.

Mycobacterium avium ssp. paratuberculosis (MAP) is the cause of Johne's disease (JD) in a wide range of domestic and wild ruminants. Single-nucleotide polymorphisms (SNPs) in several genes including solute-like carrier 11A1 (SLC11A1), interferon gamma (IFNγ), Toll-like receptor 4 (TLR4), nucleotide-binding oligomerization domain 2 gene (NOD2), and bovine peptidoglycan recognition protein 1 (PGLYRP1) have been implicated in influencing the infection outcome of MAP in cattle. We have carried out a survey in a population of Ankole cattle from three districts in the central region of Uganda including Isingiro, Lyantonde, and Rakai to determine the role played by several SNPs on the above genes in the infection outcome of local cattle in Uganda. Nine hundred fifty-five heads of cattle obtained from 93 herds were tested using ELISA. Thirty-five ELISA-positive cattle and 35 negative herd mates from a total of 955 cattle tested for MAP were genotyped using iPLEX MassARRAY genotyping systems to detect the presence of a total of 13 SNPS in five different genes (SLC11A1, IFNγ, TLR4, NOD2, and PGLYRP1). The cow-level prevalence of MAP infection in Ankole Longhorn cattle in the three districts was 3.98% (35/955), while the herd-level prevalence was 27.9% and within-herd prevalence was 12 ± 1.5% (95% CI = 9.1–14.8%). The genotypes and allele frequencies of the MAP-positive cattle were compared with those of their ELISA-negative herd mates to determine the significance of the polymorphisms. The results showed that SNPs rs109915208, rs110514940, and rs110905610 on SLC11A1, c.480G>A and c.625C>A on PGLYRP1, and c.2021C>T on TLR4 were monomorphic in both seropositive and seronegative cattle and therefore had no influence on the infection outcome. The remaining SNPs studied in the five genes [SLC11A1: rs109614179; TLR4: rs29017188 (c.226G>C), c.2021C>T; NOD2: rs110536091, rs111009394; PGLYRP1: c.102G>C, c.480G>A, c.625C>A; IFNγ: rs110853455] were polymorphic, but their allele and genotype frequencies did not show any significant difference between the seropositive and seronegative cattle. No significant difference was observed for any haplotype at the gene level.

Surveillance of Extended-Spectrum β-Lactamase-, Cephalosporinase- and Carbapenemase-Producing Gram-Negative Bacteria in Raw Milk Filters and Healthy Dairy Cattle in Three Farms in Île-de-France, France.

The aim of this work was to test a surveillance protocol able to detect extended-spectrum β-lactamase (ESBL)-, cephalosporinase (AmpC)- and carbapenemase (CP)-producing gram-negative bacteria in three conveniently chosen dairy farms with known prior occurrences of ESBL- and CP-producing strains. The protocol was applied monthly for a year. At each visit, 10 healthy lactating dairy cows were rectally swabbed, and raw milk filters (RMFs) were sampled in two of the three farms. Bacterial isolation was based on a first screening step with MacConkey agar supplemented with 1 mg/L cefotaxime and commercial carbapenem-supplemented media. We failed to detect CP-producing strains but showed that ESBL-Escherichia strains, found in one farm only (13 strains), were closely associated with multi-drug resistance (12 out of 13). The limited number of conveniently selected farms and the fact that RMFs could not be retrieved from one of them limit the validity of our findings. Still, our results illustrate that ESBL-status changes monthly based on fecal swabs and negative herds should be qualified as “unsuspected” as proposed by previous authors. Although surveillance of farm statuses based on RMF analysis could theoretically allow for a better sensitivity than individual swabs, we failed to illustrate it as both farms where RMFs could be retrieved were constantly negative. Determination of CP herd-level status based on RMFs and our surveillance protocol was hindered by the presence of intrinsically resistant bacteria or strains cumulating multiple non-CP resistance mechanisms which means our protocol is not specific enough for routine monitoring of CP in dairy farms.