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.

Comparison of diagnostic methods for assessment of Ostertagia ostertagi exposure in Norwegian dairy herds.

BACKGROUND: The gastrointestinal nematode (GIN) Ostertagia ostertagi can cause severe disease in first season grazers (FSG) and impaired performance due to subclinical infections in adult cows. Diagnostic methods to assess exposure include faecal egg count and detection of specific antibodies using antibody-ELISAs resulting in an optical density ratio (ODR). Using the ELISA test on bulk tank milk (BTM) allows for a herd level diagnosis. Appropriate use of diagnostic methods for evaluation of O. ostertagi exposure is required to optimize herd parasite surveillance and aid in a sustainable control regime. The aim of this study was to describe the relationship between different diagnostic tests used to assess GIN exposure in Norwegian production systems. A cross-sectional field study was carried out in twenty herds in Norway in the fall of 2020. Serum and faecal samples were taken from 380 individuals, of which 181 were FSG and 199 were cows. In addition, milk was collected from every cow and one BTM sample was taken from each herd. Faecal egg counts were performed. The distribution of ODR values in individual samples within and between herds and the associations between BTM ODR and individual ODR values were described. The data were analysed using visual assessment of scatter plots, Pearson correlation coefficients and linear regression. RESULTS: A high variability of the within-herd individual ODR values in serum and milk in every herd was detected. The ODR in BTM explained a low degree of the variation in the individual serum and milk samples. When plotting the ODR results in milk or serum according to four BTM categories, the distribution of ODR values were notably different in the highest and lowest BTM categories. The correlation between individual milk and serum samples was moderate (r = 0.68), while the highest correlation (r = 0.81) was between the BTM ODR and the group average individual milk samples. CONCLUSIONS: A poor predictive ability for BTM ODR to assess individual ODR values in both FSG and cows was demonstrated. However, the study indicates that the evaluation by ELISA test on BTM to assess exposure to GIN could be useful in herds with a very high or low BTM ODR.

Understanding the uptake of diagnostics for sustainable gastrointestinal nematode control by European dairy cattle farmers: a multi-country cross-sectional study.

To mitigate emerging anthelmintic resistance (AR) in cattle, sustainable gastrointestinal nematode control strategies should be adopted. A multi-centre study was set up to understand the factors affecting European dairy cattle farmers' adoption of diagnostics and to gauge for differences between regions. The data were collected through a multi-lingual survey by participating countries of the European Co-operation in Science and Technology (COST) action COMbatting Anthelmintic Resistance in ruminants (COMBAR). Four countries provided sufficient data to be included in the data analysis: Norway, Italy, Germany and Austria. Three models were estimated and validated through structural equation modelling. Norway, along with Germany and Austria (pooled dataset) showed similar trends that align with previous studies. AR risk perception had no influence on the adoption intention of diagnostics, a positive influence was found for attitude towards diagnostics and subjective norms (i.e., perceived opinion of others), and a negative influence of attitudes towards anthelminthics. Additionally, routine (i.e., perception of the current treatment) had an indirect effect on adoption intention through attitudes. Italy's data deviated from these findings, presenting a positive effect of the perceived severity of AR, and perceived behavioural control (i.e., perceived ability to perform a specific behaviour) on adoption intention of diagnostics. Finally, Norway's data set allowed for inclusion of a measurement of current behaviour in the model, identifying a direct positive effect of the perceived actual behaviour of other farmers on their own behaviour.

Title: Comprendre l'adoption des diagnostics pour le contrôle durable des nématodes gastro-intestinaux par les producteurs laitiers européens : une étude transversale multi-pays. Abstract: Pour atténuer la résistance émergente aux anthelminthiques (RA) chez les bovins, des stratégies durables de contrôle des nématodes gastro-intestinaux devraient être adoptées. Une étude multicentrique a été réalisée pour comprendre les facteurs affectant l'adoption des diagnostics par les éleveurs de bovins laitiers européens, et pour évaluer les différences entre les régions. Les données ont été recueillies au moyen d'une enquête multilingue menée par les pays participants à l'action de European Co-operation in Science and Technology (COST) COMbatting Anthelmintic Resistance in ruminants (COMBAR). Quatre pays ont collecté suffisamment de données pour être inclus dans l'analyse statistique : la Norvège, l'Italie, l'Allemagne et l'Autriche. Trois modèles ont été estimés et validés par modélisation d'équations structurelles. La Norvège, et d'autre part l'Allemagne et l'Autriche (données regroupées) ont montré des tendances similaires qui correspondent aux études précédentes. La perception de la RA n'avait aucune influence sur l'intention d'adoption des diagnostics, une influence positive a été trouvée pour l'attitude envers les diagnostics et les normes subjectives (c'est-à-dire l'opinion perçue des autres), et une influence négative des attitudes envers les anthelminthiques. De plus, la routine (c'est-à-dire la perception du traitement actuel) a eu un effet indirect sur l'intention d'adoption par le biais des attitudes. Les données de l'Italie s'écartaient des résultats précédents, présentant un effet positif de la gravité perçue du risque de la RA, et du contrôle comportemental perçu (c'est-à-dire la capacité perçue à adopter un comportement spécifique) sur l'intention d'adoption de diagnostic. Enfin, l'ensemble de données de la Norvège a permis d'inclure une mesure du comportement actuel dans le modèle, identifiant un effet positif direct du comportement réel perçu des autres agriculteurs sur son propre comportement.

Gastrointestinal nematodes and Fasciola hepatica in Norwegian cattle herds: a questionnaire to investigate farmers' perceptions and control strategies.

BACKGROUND: Pasture management influences the prevalence and impact of the pasture parasites (PP) in cattle herds, which cause production-limiting disease worldwide. Evaluating farmer management strategies is vital when considering sustainable PP control practices. The aim of this questionnaire-based study was to describe the pasture management and control strategies regarding PP in Norwegian beef cattle (BC) and dairy cattle (DC) production systems with a focus on gastrointestinal nematodes (GIN) and Fasciola hepatica. RESULTS: A total of 745 responses from BC (return rate 20.5%) and 1347 responses from DC farmers (30.7%) were included. The mean total pasture time for DC was 4.2 months for first-season grazers and 4.3 months for second-season grazers and cows, while the corresponding finding in BC was 5.4 months. Home pasture was used for most of the pasture period, particularly for first-season grazer dairy heifers (81%), which were also commonly grazed on the same pasture every year (79%). For most farmers it was necessary for grazing areas to be used for cattle for more than one season (77% of BC farmers and 89% of DC farmers). However, changing the pasture during the season was common in both DC (67%) and BC (70%) herds. The majority of DC farmers (60%) stated that they did not consider that they had a problem with PP. Of the remaining 40%, few respondents could specify whether their herds had a problem due to infection by GIN (11%) or liver flukes (12%). Treatment for GIN was performed by 52% of DC and 34% of BC farmers. Diagnostic faecal samples were collected upon suspicion of parasitic disease by 5% of DC and 16% of BC farmers. Veterinarians were stated as a central source of information about parasite management and treatment. CONCLUSIONS: Potential risks for exposure to PP were identified, such as use of the same pasture every year for first-season grazers and frequent use of home pasture. The perception of problems related to PP appeared low. Regular anthelmintic treatment without concurrent use of diagnostic faecal samples seems to be common practice.

Field evaluation of anticoccidial efficacy: A novel approach demonstrates reduced efficacy of toltrazuril against ovine Eimeria spp. in Norway.

Ovine Eimeria spp. infections cause reduced welfare, increased mortality, and substantial economic losses, and anticoccidials are crucial for their control. Recent reports of toltrazuril resistance in pigs, and anecdotal reports of reduced anticoccidial efficacy in lambs, necessitate evaluation of anticoccidial efficacy. Due to the substantial lifecycle differences between nematodes and coccidia, current WAAVP methods for assessing anthelmintic efficacy are not suitable for such evaluations. Faecal samples were collected from 8 pairs of twin lambs from 36 Norwegian sheep farms 6-8 days after turnout. One twin of each pair was then treated with 20 mg/kg toltrazuril and a second faecal sample from all lambs was collected 7-11 days later. Oocyst excretion rate in all samples was determined using McMasters. Suitability of treatment timing was investigated by evaluating the increase in mean log oocyst excretion in untreated lambs. Based on comparisons between groups, a threshold of ≥0.75 (13 farms) was used to identify farms where drug efficacy could be assessed with confidence, drug efficacy on farms with increases of ≥0.5 but <0.75 (7 farms) were evaluated with caution, and drug efficacy on farms with increases of <0.5 (16 farms) was not estimated. Reduction in oocyst excretion between samples from treated lambs compared with controls from the 20 farms with a threshold of ≥0.5 were then analysed using a generalised linear mixed model. The results were classified based on 95% CI obtained using parametric bootstrapping. Among these 20 farms, two exhibited reduced drug efficacy (upper 95% CI < 95%), 13 had good efficacy (lower 95% CI > 90%), and for 5 the results were inconclusive. This is the first evidence-based report of reduced anticoccidial efficacy in ovine Eimeria spp. Additionally, we highlight the problem of sub-optimal timing of treatment (16/36 farms), which could potentially result in incorrect conclusions being reached regarding lack of drug efficacy.

Treatment against coccidiosis in Norwegian lambs and potential risk factors for development of anticoccidial resistance-a questionnaire-based study.

The objectives of this study were to investigate the use of anticoccidials in Norwegian sheep flocks and identify farms with management procedures likely to select for drug resistance. Data were obtained by a questionnaire sent to all members of the Norwegian Sheep Recording System in October 2015. The data set consisted of 1215 answers, corresponding to 8.5% of Norwegian sheep flocks. Anticoccidials were used in 82.7% of flocks. Main treatment was at turnout (38.6% of treated flocks) or 1 week after turnout (32.4%). Interestingly, clinical signs possibly related to coccidiosis were observed by almost 40% of the farmers after treatment, which might be an indication of drug resistance. Correlations between the apparently reduced anticoccidial efficacy and management conditions, such as the size of the farms, were found. From the farmers' perspective, metaphylactic treatment was used in 88.5% of treated flocks, of which approximately one third had no history of clinical coccidiosis. Even though farmers seem aware of the importance of good drenching routines based on reliable estimates of weights and calibration of drench guns, drench gun used for anticoccidial administration was never calibrated in 12.1% of the flocks. Finally, dose estimation was made by visual appraisal in 27.5% of the flocks, which can lead to incorrect dosing. Based on the present study, it cannot be determined whether the apparent treatment failure was related to management practises, incorrect administration of the drug, other infections or actual anticoccidial drug resistance.

Experimental infection of cattle with ovine Dichelobacter nodosus isolates.

BACKGROUND: Dichelobacter nodosus is the main causative agent of ovine footrot, and there are strong indications that the bacterium can be transferred to cattle grazing on the same pasture as sheep. The aim of this study was to investigate if benign and virulent D. nodosus strains isolated from sheep can be transferred to the interdigital skin of cattle under experimental conditions. Further, we wanted to observe the impact of such infection on bovine foot health, and test the effect of topical chlortetracycline (Cyclo spray(®): Eurovet) on the infection. FINDINGS: Six heifers were included in the study. After an initial 18-day maceration period, three heifers were inoculated on one single foot with a benign strain and three with a virulent strain by adding bacterial suspension in a bandage. The bandages were left on for 17 days, and when removed, D. nodosus was isolated from all six heifers. All six heifers developed interdigital dermatitis. In five of the heifers D. nodosus organisms were demonstrated within the epidermis. Twenty-four days after treatment with chlortetracycline all heifers were negative by cultivation, but tested positive for D. nodosus by polymerase chain reaction (PCR). Two of the six heifers still tested positive for D. nodosus by PCR 49 days after treatment. After 70 days, all heifers tested negative for D. nodosus. CONCLUSIONS: This study shows that both virulent and benign D. nodosus strains originating from sheep can be transferred to naïve heifers under experimental conditions. Further, the study supports the hypothesis that infections with virulent D. nodosus in cattle are associated with interdigital dermatitis. No conclusion regarding the treatment of D. nodosus infection with chlortetracycline was possible.

Housing conditions and management practices associated with neonatal lamb mortality in sheep flocks in Norway.

A study was conducted in order to obtain information about sheep farms in Norway and to identify housing and management characteristics that were risk factors for neonatal mortality of lambs 0-5 days of age. A questionnaire was submitted to sheep farmers, who provided demographic data and information on sheep housing conditions and feeding and management practices. Our description of farms is based on the questionnaire responses received from 2260 farmers. Data on lamb mortality during the preceding lambing season were available for those flocks that were enrolled in the Norwegian Sheep Recording System. Some flocks where the number of lambing ewes was less than 20 or greater than 400 were excluded. The total number of flocks included in the analysis of neonatal mortality was 1125. An increase in the mean number of live-born lambs per ewe per flock was associated with increasing neonatal mortality. Factors independently associated with increased neonatal survival were continuous monitoring of the ewes during the lambing season, active support to ensure sufficient colostrum intake of the lambs, feeding a combination of grass silage and hay compared with grass silage alone, and supplying roughage at least twice per day versus only once. Increased survival was also observed in flocks where the farmer had at least 15 years of experience in sheep farming. Flocks in which the Spæl breed predominated had lower odds for neonatal deaths compared to flocks in which the Norwegian White breed predominated. In conclusion, measures in sheep flocks targeted at feeding practices during the indoor feeding period and management practice during lambing season would be expected to reduce neonatal lamb mortality.

Possible cross-infection of Dichelobacter nodosus between co-grazing sheep and cattle.

BACKGROUND: The aim of this study was to investigate possible cross-infection of Dichelobacter nodosus in Norwegian farms practising co-grazing of sheep and cattle. METHODS: Thirteen farms practising co-grazing of sheep and cattle were included in this descriptive study: five farms with a history of severe ovine footrot (Group I) and eight farms with free-stall housing of cattle and signs of mild or no footrot in sheep (Group II). Sampling for PCR detection of D. nodosus was performed from animals in all farms, and clinical claw examination of sheep and cattle was performed in Group II. D. nodosus positive samples were analysed by a multiplex PCR method that detects variants of the fimA gene corresponding to D. nodosus serogroups A through I. RESULTS: D. nodosus serogroup A was identified more frequently in sheep from farms with a history of severe footrot (Group I) versus from Group II, and in most of the farms with a history of severe footrot there was a coexistence of D. nodosus serogroup A in sheep and cattle. In one farm heel horn erosion and dermatitis emerged in cattle after co-grazing with sheep suffering from severe footrot where D. nodosus serogroup A was detected. Six months later heel horn erosion and dermatitis were still diagnosed, and D. nodosus serogroup A was identified. Out of the 16 D. nodosus positive sheep samples from Group II, ten of the samples were positive by the fimA serogrouping PCR. Among these 10 samples all serogroups except G were detected. All the D. nodosus serogroups detected in sheep were also present in the corresponding cattle herds. CONCLUSION: The clinical findings and the coexistence of the same serogroups in co-grazing sheep and cattle could indicate cross-infection. However, further research including isolation of the bacterial strains, virulence-testing and genetic identification, is needed.