Plant-based production of a protective vaccine antigen against the bovine parasitic nematode Ostertagia ostertagi.

The development of effective recombinant vaccines against parasitic nematodes has been challenging and so far mostly unsuccessful. This has also been the case for Ostertagia ostertagi, an economically important abomasal nematode in cattle, applying recombinant versions of the protective native activation-associated secreted proteins (ASP). To gain insight in key elements required to trigger a protective immune response, the protein structure and N-glycosylation of the native ASP and a non-protective Pichia pastoris recombinant ASP were compared. Both antigens had a highly comparable protein structure, but different N-glycan composition. After mimicking the native ASP N-glycosylation via the expression in Nicotiana benthamiana plants, immunisation of calves with these plant-produced recombinants resulted in a significant reduction of 39% in parasite egg output, comparable to the protective efficacy of the native antigen. This study provides a valuable workflow for the development of recombinant vaccines against other parasitic nematodes.

Alleviating lesions of chronic progressive lymphedema in Belgian draft horses by successfully treating Chorioptes bovis infestation with moxidectin 0.5% pour-on.

The aim of this prospective, randomized, single-blinded, and placebo-controlled clinical trial was to investigate the efficacy of a moxidectin pour-on solution for the treatment of Chorioptes bovis infestation in Belgian draft horses, and in addition, to evaluate the effect of this treatment on the clinical signs and lesions associated with chronic progressive lymphedema (CPL). Nineteen privately owned Belgian draft horses were randomly assigned to either a treatment group (moxidectin pour-on formulation, n = 10) or a placebo group (phosphate-buffered saline (PBS), n = 9). On Day 0, all 19 horses tested positive for the presence of C. bovis in superficial skin scrapings. Prior to treatment, all feathering on the distal limbs of the horses was clipped. Treatment was applied twice (Day 0 and 7). Pour-on moxidectin (Cydectin 0.5% Pour-On; Zoetis) was evenly distributed over the distal legs of the horses at a dose of 1.5 mg moxidectin/kg body weight. Animals in the placebo group were treated with PBS. Pretreatment and follow-up examinations consisted of counting living mites in superficial skin scrapings, scoring pruritus, and scoring mange-associated and CPL-associated lesions (skinfold score and skin lesion score). Horses in the placebo group and moxidectin group were followed up to 8 weeks and 24 weeks after the first treatment, respectively. On Day 14, no living mites were found in any of the horses in the moxidectin group (p = 0.013). These horses continued to remain free of mites, until the final sampling conducted at 24 weeks following the initial application of moxidectin, when three horses again showed living mites in skin scrapings. Treatment with moxidectin resulted in a significant reduction of both CPL-associated skin lesion scores (p = 0.003) and pruritus scores (p = 0.001) after only seven days. By Day 56, still no signs of pruritus (p < 0.0001) were detected, with significant improvement of mange-associated lesions (p < 0.0001). Although the skinfold score did not show a significant reduction by Day 56, the score for skin lesions associated with CPL had significantly improved (p < 0.0001). In conclusion, the results of this study demonstrate that pour-on moxidectin, at a high dose and applied directly to the mite predilection site, was an effective treatment for C. bovis infestation in feathered draft horses, providing positive effects on CPL lesions, pruritus and mange-associated lesions. Furthermore, these findings emphasize the therapeutic significance of addressing mange in the management of CPL-affected draft horses.

Review and Evaluation of Ostertagia ostertagi Antibody ELISA for Application on Serum Samples in First Season Grazing Calves.

The O. ostertagi-Ab ELISA assay is widely used as a diagnostic tool for monitoring gastrointestinal (GI) nematodes using milk samples from adult dairy cows. This assay is potentially also useful to analyse serum samples from first-season grazing (FSG) calves, providing a more cost-effective and robust diagnostic technique than the current serum pepsinogen assay. However, a comprehensive evaluation of its use in serum samples from FSG calves has not yet been conducted. In this study, we first reviewed the available scientific literature in which the O. ostertagi-Ab ELISA was applied to serum samples from FSG calves. Then, a field study was conducted to compare results from the O. ostertagi-Ab ELISA assay with a serum pepsinogen assay on a set of 230 serum samples from 11 commercial dairy herds (seven in Belgium and four in Sweden). The literature review showed an increase in mean antibody levels, expressed as optical density ratio (ODR) values, from <0.4 (early grazing season) to values of 0.7-1.1 (late grazing season). Three out of five studies found a negative correlation between O. ostertagi antibody levels measured during the late grazing season and weight gain, while the other two studies found no correlation between the two variables. Our field studies showed a good correlation between O. ostertagi antibody levels and the results from the pepsinogen assay. Both indicators were negatively related to average daily weight gain in the Belgian herds, but not in the Swedish herds. Overall, the results suggest that the O. ostertagi-Ab ELISA test can be a useful tool in FSG calves and could replace the use of the serum pepsinogen assay at the end of the grazing season for general monitoring purposes.

Chronic Progressive Lymphedema in Belgian Draft Horses: Understanding and Managing a Challenging Disease.

Chronic progressive lymphedema (CPL) in draft horses is characterized by increased dermal thickness and fibrosis, with the development of skinfolds and nodules, hyperkeratosis, and ulcerations on the distal limbs of affected horses. Secondary bacterial, fungal, or parasitic infections frequently complicate and aggravate the lesions, as well as the progression of this disease. CPL has a particularly high prevalence of up to 85.86% in the Belgian draft horse breed. Due to the disease's progressive and incurable nature, affected horses are often euthanized prematurely. The treatment options are solely symptomatic, aimed at improving the horse's quality of life. Despite the severity of this condition, many uncertainties about its etiology and pathogenesis still remain to date. The established scientific research on CPL is rather limited, although there is an urgent need for strategies to tackle this disease. This review summarizes the available knowledge, serving as a guideline for practitioners, and provides perspectives for future research programs.

Practical guide to the diagnostics of ruminant gastrointestinal nematodes, liver fluke and lungworm infection: interpretation and usability of results.

The diagnostics of ruminant parasites remains one of the cornerstones for parasite control best practices. Field veterinarians have several techniques at their disposal (fecal egg count, coproculture, FAMACHA®, plasma pepsinogen, ELISA-Ostertagia, ELISA-Fasciola, Baermann and ELISA-Lungworm) for the identification and/or quantification of gastrointestinal nematodes, lungworms and liver fluke infecting small ruminants and cattle. Each of these diagnostic tools has its own strengths and weaknesses and is more appropriate for a specific production operation and/or age of the animal (young and adults). This review focuses on the usability and interpretation of the results of these diagnostic tools. The most advanced technical information on sampling, storage, advantages and limitations of each tool for different types of production operations and animal categories is provided.

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.

To treat or not to treat: diagnostic thresholds in subclinical helminth infections of cattle.

Helminth infections of cattle place significant burdens on livestock production and farm economic efficiency. Heavy infections are relatively easy to detect and treat with anthelmintics. However, subclinical infections have major but often hidden impacts on animals, necessitating more refined diagnostics to detect them and ideally inform farmers about the likely impact of anthelmintic treatment on animal and herd performance. Here, we review recent advances in diagnosing three major cattle helminth infections - gastrointestinal nematodes (GINs), liver flukes, and lungworms - and the search for subclinical infection thresholds to guide treatment decisions. Combining refined diagnostic thresholds with farm-specific information on grazing systems and animal history enables farmers to tailor helminth treatments to specific epidemiological circumstances, thereby limiting anthelmintic resistance (AR) and boosting agricultural efficiency and food security.

Exploration of the Ixodes ricinus virosphere unveils an extensive virus diversity including novel coltiviruses and other reoviruses.

Recent metagenomics studies have revealed several tick species to host a variety of previously undiscovered RNA viruses. Ixodes ricinus, which is known to be a vector for many viral, bacterial, and protozoan pathogens, is the most prevalent tick species in Europe. For this study, we decided to investigate the virosphere of Belgian I. ricinus ticks. High-throughput sequencing of tick pools collected from six different sampling sites revealed the presence of viruses belonging to many different viral orders and families, including Mononegavirales, Bunyavirales, Partitiviridae, and Reoviridae. Of particular interest was the detection of several new reoviruses, two of which cluster together with members of the genus Coltivirus. This includes a new strain of Eyach virus, a known causative agent of tick-borne encephalitis. All genome segments of this new strain are highly similar to those of previously published Eyach virus genomes, except for the fourth segment, encoding VP4, which is markedly more dissimilar, potentially indicating the occurrence of a genetic reassortment. Further polymerase chain reaction-based screening of over 230 tick pools for 14 selected viruses showed that most viruses could be found in all six sampling sites, indicating the wide spread of these viruses throughout the Belgian tick population. Taken together, these results illustrate the role of ticks as important virus reservoirs, highlighting the need for adequate tick control measures.

Three-year study to evaluate an anthelmintic treatment regimen with reduced treatment frequency in horses on two study sites in Belgium.

In the present study, an anthelmintic treatment regimen with reduced treatment frequency was evaluated in horses on two study sites in Belgium during three consecutive summer pasture seasons. Historically, the horses on both study sites were treated up to 6 times a year with ivermectin (IVM) or up to 4 times a year with moxidectin (MOX), and previous efficacy evaluations indicated a reduced egg reappearance period in some of the treated horses for both IVM (28 days) and MOX (42 days). In the present study, all horses were treated with IVM or MOX in the spring and in autumn. Faecal egg counts (FEC) were conducted every two weeks during the summer pasture season and whenever the individual FEC exceeded 250 eggs per gram of faeces, the specific horse was treated with pyrantel embonate. No increase in parasitic disease over the three-year period of the study was observed. The FEC data collected in the study as well as the age of the animals and local weather data were then imported into a cyathostomin life-cycle model, to evaluate long term effects of the newly applied treatment regimen on the selection pressure for anthelmintic resistance, and compare to the previous high frequency treatment regimen. The model simulations indicated that the whole-herd treatment regimen with at least 4 macrocyclic lactone treatments annually led 2-3 times faster resistance development than any of the alternative treatment regimens evaluated under the specific conditions of these two study sites. Further lowering the treatment frequency or applying even more selective treatments enhanced the delay in resistance development, but to a lesser extent.

Subclinical nematode parasitism affects activity and rumination patterns in first-season grazing cattle.

Sickness behaviour has been suggested as an applicable indicator for monitoring disease. Deviating feeding behaviour and activity can provide information about animals' health and welfare status. Recent advances in sensor technology enable monitoring of such behaviours and could potentially be utilized as an indicator of gastrointestinal nematode (GIN) infections. This study investigated activity and rumination patterns in first-season grazing steers exposed to subclinical infection levels of the GIN Ostertagia ostertagi and Cooperia oncophora. At turnout, animals were allocated to one of four experimental groups and were faced with "high" (H1, n = 15; H2, n = 17) or "low" (L1, n = 17; L2, n = 11) levels of parasite exposure by grazing in similar enclosures contaminated with overwintering third stage (L3) GIN larvae. Animals in H1 and H2 (HP) received a 1:1 mix of approximately 10,000 O. ostertagi and C. oncophora L3 at turnout; whereas the animals in L1 and L2 (LP) were treated monthly with ivermectin. Activity and rumination patterns were monitored by fitting animals with leg- (IceQube) and neck-mounted (Heatime) sensors. BW was recorded every fortnight, whereas faecal and blood samples were examined every four weeks for nematode faecal egg count and serum pepsinogen concentrations (SPCs). There was an interaction effect of exposure level and period (P < 0.0001) on average lying daily time across the entire grazing time. A higher mean daily lying time (P = 0.0037) was found in HP compared with LP during the first 40 days on pasture. There was also interaction effects of treatment and day since turnout on rumination time (P < 0.0001) and rumination change (P = 0.0008). Also mean daily steps (P < 0.0001) and mean daily motion index (P < 0.0001) were markedly higher in HP during days 62-69, coinciding with peaking SPC in HP. Strongyle eggs were observed both in HP and LP from 31 days after turnout. Eggs per gram (EPG) differed between parasite exposure levels (P < 0.0001), with mean EPG remaining low in LP throughout the experiment. Similarly, an increase in SPC was observed (P < 0.0001), but only in HP where it peaked at day 56. In contrast, no difference in BW gain (BWG) (P = 0.78) between HP and LP was observed. In conclusion, this study shows that behavioural measurements monitored with sensors were affected even at low infection levels not affecting BWG. These combined results demonstrate the potential of automated behavioural recordings as a tool for detection of subclinical parasitism.

Constraints of using historical data for modelling the spatial distribution of helminth parasites in ruminants.

Dicrocoelium dendriticum is a trematode that infects ruminant livestock and requires two different intermediate hosts to complete its lifecycle. Modelling the spatial distribution of this parasite can help to improve its management in higher risk regions. The aim of this research was to assess the constraints of using historical data sets when modelling the spatial distribution of helminth parasites in ruminants. A parasitological data set provided by CREMOPAR (Napoli, Italy) and covering most of Italy was used in this paper. A baseline model (Random Forest, VECMAP®) using the entire data set was first used to determine the minimal number of data points needed to build a stable model. Then, annual distribution models were computed and compared with the baseline model. The best prediction rate and statistical output were obtained for 2012 and the worst for 2016, even though the sample size of the former was significantly smaller than the latter. We discuss how this may be explained by the fact that in 2012, the samples were more evenly geographically distributed, whilst in 2016 most of the data were strongly clustered. It is concluded that the spatial distribution of the input data appears to be more important than the actual sample size when computing species distribution models. This is often a major issue when using historical data to develop spatial models. Such data sets often include sampling biases and large geographical gaps. If this bias is not corrected, the spatial distribution model outputs may display the sampling effort rather than the real species distribution.

TITLE: Contraintes liées à l'utilisation de données historiques pour la modélisation de la distribution spatiale des helminthes parasites chez les ruminants. ABSTRACT: Dicrocoelium dendriticum est un trématode qui infecte les ruminants et nécessite deux hôtes intermédiaires différents pour terminer son cycle de vie. La modélisation de la distribution spatiale de ce parasite peut aider à améliorer sa gestion dans les régions à haut risque. L'objectif de cette recherche était d'évaluer les contraintes liées à l'utilisation d'ensembles de données historiques lors de la modélisation de la distribution spatiale des helminthes parasites chez les ruminants. Un ensemble de données parasitologiques fourni par CREMOPAR (Naples, Italie) et couvrant la majeure partie de l'Italie a été utilisé dans cet article. Un modèle de base (Random Forest, VECMAP®) utilisant l'ensemble des données a d'abord été utilisé pour déterminer le nombre minimal de points de données nécessaires pour construire un modèle stable. Ensuite, des modèles de distribution annuelle ont été calculés et comparés au modèle de référence. Le meilleur taux de prédiction et le meilleur résultat statistique ont été obtenus pour 2012 et le plus mauvais pour 2016, malgré le fait que la taille de l'échantillon du premier était nettement plus petite que celle du second. Nous discutons comment cela peut s'expliquer par le fait qu'en 2012, les échantillons étaient plus uniformément répartis géographiquement, alors qu'en 2016, la plupart des données étaient fortement regroupées. On conclut que la distribution spatiale des données d'entrée semble être plus importante que la taille réelle de l'échantillon lors du calcul des modèles de distribution des espèces. C'est souvent un problème majeur pour développer des modèles spatiaux quand on utilise des données historiques. Ces ensembles de données comportent souvent des biais d'échantillonnage et de grandes lacunes géographiques. Si ce biais n'est pas corrigé, les résultats du modèle de distribution spatiale peuvent représenter l'effort d'échantillonnage plutôt que la distribution réelle des espèces.

Unravelling Belgian Blue cattle farmers' adoption intention towards diagnostic tools: Integrating insights from behavioural economics and socio-cognitive theories.

Macrocyclic lactone resistance in Psoroptes ovis mites, causing psoroptic mange, is emerging in beef cattle. Therefore, diagnostic tools that can indicate macrocyclic lactone resistance should be implemented at farms in order to control these parasitic infections and slow down emerging resistance. Unfortunately, the adoption of such tools remains relatively poor. This study explores which beliefs, and behavioural biases that misconstrue these beliefs, underlie farmers' adoption intention. Five semi-structured focus groups were conducted with Belgian Blue cattle farmers (n = 38). Inductive analysis revealed that farmers' positive attitudes towards acaricides use and risk perception towards acaricide resistance made them rather reluctant to implement diagnostic tools. In addition, farmers' susceptibility to acaricide resistance occurring on their farm was weakened by the optimism and availability bias. Deductive analysis revealed that economic reasons, usefulness of the diagnostic tool and contribution to animal wellbeing motivated farmers to adopt such tools (i.e., behavioural beliefs). However, the loss aversion and time discounting bias weakened farmers' beliefs related to economic reasons. The veterinarian was seen as the responsible actor for implementing diagnostic tools, while colleague-farmers' opinion related to the diagnostic tool was also valued (i.e., normative beliefs). The latter belief was strengthened by the bandwagon-effect bias. Farmers' beliefs about economic costs related to the diagnostic tool tended to hinder adoption intention, while the veterinarian's assistance motivated farmers to implement such tools (i.e., control beliefs). The loss aversion bias also strengthened farmers' control beliefs related to the diagnostic tools' perceived costs. As such, this article provides more insights into the (ir)rational factors shaping farmers' intention to adopt diagnostic tools. These insights might help animal health organisations to design communication strategies to stimulate the adoption of diagnostic tools on beef cattle farms.

Dermal immune responses against Psoroptes ovis in two cattle breeds and effects of anti-inflammatory dexamethasone treatment on the development of psoroptic mange.

Psoroptic mange is a common disease of livestock, caused by Psoroptes ovis. Compared to Holstein-Friesian (HF) cattle, the Belgian Blue (BB) cattle breed is highly susceptible to the infestation. However, the mechanism for this difference is still unclear. To determine the factors responsible for this breed susceptibility, the immune response to P. ovis was studied in experimentally infested BB and HF cattle, using clinical signs, histology, immunohistochemical profiling and gene expression analysis of skin biopsies. The mite numbers and lesion area of BB cattle were greater than in HF during the whole study period. Significant influxes of eosinophils in the epidermis and dermis were detected in comparison with the pre-infestation samples in both breeds, with significantly higher eosinophils in BB at 6 weeks post infestation (wpi). Mast cell numbers were unaffected at all stages of infestation in HF, but were significantly elevated relative to pre-infestation in BB cattle at 2 and 6 wpi. The more pronounced cutaneous eosinophilia and higher IL-4 levels at 6 wpi in BB cattle suggest that a Th2-type immune response is underlying the higher susceptibility of the BB breed. In naturally infested BB cattle, development of the psoroptic mange lesions and eosinophils and CD3+ T cell areas were severely depressed after anti-inflammatory treatment with dexamethasone. Together, these results suggest that a stronger Th2-type immune response to P. ovis causes the skin lesions in psoroptic mange in BB cattle and that local anti-inflammatory treatment could potentially be an alternative to control the pathology caused by this parasite.

Increasing importance of anthelmintic resistance in European livestock: creation and meta-analysis of an open database.

Helminth infections are ubiquitous in grazing ruminant production systems, and are responsible for significant costs and production losses. Anthelmintic Resistance (AR) in parasites is now widespread throughout Europe, although there are still gaps in our knowledge in some regions and countries. AR is a major threat to the sustainability of modern ruminant livestock production, resulting in reduced productivity, compromised animal health and welfare, and increased greenhouse gas emissions through increased parasitism and farm inputs. A better understanding of the extent of AR in Europe is needed to develop and advocate more sustainable parasite control approaches. A database of European published and unpublished AR research on gastrointestinal nematodes (GIN) and liver fluke (Fasciola hepatica) was collated by members of the European COST Action "COMBAR" (Combatting Anthelmintic Resistance in Ruminants), and combined with data from a previous systematic review of AR in GIN. A total of 197 publications on AR in GIN were available for analysis, representing 535 studies in 22 countries and spanning the period 1980-2020. Reports of AR were present throughout the European continent and some reports indicated high within-country prevalence. Heuristic sample size-weighted estimates of European AR prevalence over the whole study period, stratified by anthelmintic class, varied between 0 and 48%. Estimated regional (country) prevalence was highly heterogeneous, ranging between 0% and 100% depending on livestock sector and anthelmintic class, and generally increased with increasing research effort in a country. In the few countries with adequate longitudinal data, there was a tendency towards increasing AR over time for all anthelmintic classes in GIN: aggregated results in sheep and goats since 2010 reveal an average prevalence of resistance to benzimidazoles (BZ) of 86%, macrocyclic lactones except moxidectin (ML) 52%, levamisole (LEV) 48%, and moxidectin (MOX) 21%. All major GIN genera survived treatment in various studies. In cattle, prevalence of AR varied between anthelmintic classes from 0-100% (BZ and ML), 0-17% (LEV) and 0-73% (MOX), and both Cooperia and Ostertagia survived treatment. Suspected AR in F. hepatica was reported in 21 studies spanning 6 countries. For GIN and particularly F. hepatica, there was a bias towards preferential sampling of individual farms with suspected AR, and research effort was biased towards Western Europe and particularly the United Kingdom. Ongoing capture of future results in the live database, efforts to avoid bias in farm recruitment, more accurate tests for AR, and stronger appreciation of the importance of AR among the agricultural industry and policy makers, will support more sophisticated analyses of factors contributing to AR and effective strategies to slow its spread.

TITLE: Importance croissante de la résistance aux anthelminthiques chez les ruminants européens : création et méta-analyse d'une base de données ouverte. ABSTRACT: Les helminthes sont omniprésents dans les systèmes de production de ruminants au pâturage et sont responsables de coûts et de pertes de production importants. La résistance aux anthelminthiques (RA) des parasites est maintenant répandue dans toute l'Europe, bien qu'il existe encore des lacunes dans nos connaissances dans certaines régions et certains pays. La RA est une menace majeure pour la pérennité de la production moderne de ruminants, en diminuant la productivité, en compromettant la santé et le bien-être des animaux, et en augmentant les émissions de gaz à effet de serre au travers d'une augmentation du parasitisme et des intrants agricoles. Une meilleure compréhension de l'étendue de la RA en Europe est nécessaire pour développer et préconiser des approches de lutte antiparasitaire plus durables. Une base de données intégrant des informations publiées et non publiées en Europe concernant la RA des nématodes gastro-intestinaux (NGI) et de la douve du foie (Fasciola hepatica) a été compilée par les membres de l'action européenne COST « COMBAR ¼ (« Combattre la résistance aux anthelminthiques chez les ruminants ¼) et combinée avec les données d'une précédente étude systématique concernant la RA des NGI. Au total, 197 publications sur la RA des NGI étaient disponibles pour analyse, représentant 535 études dans 22 pays et couvrant la période 1980­2020. Des signalements de RA étaient présents sur tout le continent européen et certains rapports indiquaient une forte prévalence nationale. Les estimations heuristiques pondérées par la taille de l'échantillon de la prévalence de la RA en Europe sur toute la période d'étude, stratifiées par classe d'anthelminthiques, variaient de 0 à 48 %. La prévalence régionale (nationale) estimée était très hétérogène, variant entre 0 % et 100 % selon le secteur de l'élevage et la classe d'anthelminthique, et augmentait généralement avec les efforts de recherche dans le pays. Dans les quelques pays disposant de données longitudinales adéquates, il y avait une tendance à l'augmentation de la RA au fil du temps pour toutes les classes d'anthelminthiques des NGI : les résultats agrégés chez les ovins et caprins depuis 2010 révèlent une prévalence moyenne de résistance aux benzimidazoles (BZ) de 86 %, aux lactones macrocycliques sauf moxidectine (ML) de 52 %, au lévamisole (LEV) de 48 % et à la moxidectine (MOX) de 21 %. Tous les genres principaux de NGI ont survécu au traitement dans diverses études. Chez les bovins, la prévalence de la RA variait selon les classes d'anthelminthiques de 0 à 100 % (BZ et ML), 0 à 17 % (LEV) et 0 à 73 % (MOX), et Cooperia et Ostertagia ont survécu aux traitements. Une RA suspectée chez F. hepatica a été signalée dans 21 études portant sur 6 pays. Pour les NGI, et encore plus pour F. hepatica, il y avait un biais d'échantillonnage en faveur des exploitations individuelles suspectées de RA, et l'effort de recherche était biaisé vers l'Europe occidentale et en particulier le Royaume-Uni. La capture continue des résultats futurs dans la base de données, en direct, les efforts pour éviter les biais dans le recrutement des exploitations, des tests plus précis pour la RA et une meilleure appréciation de l'importance de la RA parmi l'industrie agricole et les décideurs politiques, soutiendront des analyses plus sophistiquées des facteurs contribuant à la RA, et des stratégies efficaces pour ralentir sa propagation.

A Qualitative Market Analysis Applied to Mini-FLOTAC and Fill-FLOTAC for Diagnosis of Helminth Infections in Ruminants.

Helminth infections, mainly by gastrointestinal nematodes (GIN), are one of the main concerns for animal health, welfare and productivity in grazing ruminant livestock worldwide. The use of a sensitive, precise, accurate, low-cost, and easy-to-perform copromicroscopic technique is of pivotal importance to perform reliable fecal egg count (FEC) and fecal egg count reduction test (FECRT), in order to determine the need of anthelmintic treatment, but also anthelmintic efficacy or resistance. This approach is fundamental to a correct and efficient control of GIN. Unfortunately, in worldwide ruminant farm practice, repeated anthelmintic treatments are carried out, without prior diagnosis of infection, contributing to the spread of Anthelmintic Resistance (AR). Tackling this phenomenon, improving mainly the GIN diagnosis and AR status in farm animals, is a priority of the European COST Action "COMBAR-COMBatting Anthelmintic Resistance in Ruminants" and of the STAR-IDAZ International Research Consortium on Animal Health. One of the specific objectives of the COMBAR Working Group 1 (WG1) is to conduct an European market analysis of new diagnostics and develop a business plan for commercial test introduction, leveraging technical know-how of participants. Since the Mini-FLOTAC in combination with the Fill-FLOTAC may be considered a good candidate for a standardized FEC and FECRT in the laboratory, as well as directly in the field, the aim of this study was to conduct SWOT (Strength-Weaknesses-Opportunities-Threats) and PESTEL (Political, Economic, Social, Technological, Environmental, and Legal) analyses of these tools in 20 European countries involved in the COMBAR WG1, in order to identify the opportunities, barriers, and challenges that might affect the Mini-FLOTAC and Fill-FLOTAC commercialization in Europe.

New insights into the use of a mite count reduction test for the detection of therapeutic acaricide efficacy in Psoroptes ovis in cattle.

When used for the evaluation of drug efficacy against Psoroptes ovis, the diagnostic performance of different sampling strategies for a mite count reduction test (MCRT) remains unclear. In the present study, a novel simulation framework was constructed that accounted for relevant biological features of P. ovis infestations in cattle and that was parameterized with field data from 16 farms (154 animals). Second, this framework was applied to explore the impact of study specific factors (number of animals, number of sampled lesions, and number of scrapings per lesion) and biological factors (mite infestation intensity and size of lesions) on the diagnostic performance of MCRT. Its outcome provided a basis to determine the diagnostic performance of MCRT when it was applied according to the World Association for the Advancement in Veterinary Parasitology (WAAVP) and the European Medicine Agency (EMA) guidelines, and to formulate recommendations to ensure a good diagnostic performance of the MCRT. For both guidelines, the MCRT allowed to correctly detect (power 80%) reduced and normal efficacy when the therapeutic efficacy was <70%, and ≥95%, respectively. The results highlighted a reliable diagnostic performance of the MCRT when performed as recommended by WAAVP and EMA for the detection of normal drug efficacy. When used for the detection of reduced efficacy, therapeutic efficacies between 70% and 90% could not be detected with sufficient reliability. The diagnostic performance can be improved by increasing the total number of skin scrapings (increasing the number of animals, number of sampled lesions and/or number of samples per lesion). In order to help researchers and veterinarians to optimize the design of the MCRT to their field settings, the findings were translated into a simple tool.

Cattle gastrointestinal nematode egg-spiked faecal samples: high recovery rates using the Mini-FLOTAC technique.

BACKGROUND: Faecal egg count (FEC) techniques are commonly used to detect gastrointestinal nematodes (GINs) in cattle and to determine anthelmintic efficacy/resistance through the faecal egg count reduction test (FECRT). Mini-FLOTAC is one of the techniques recommended for a standardised FEC/FECRT of helminth eggs in cattle. However, only one paper evaluated the recovery rate of GIN eggs by Mini-FLOTAC (compared to McMaster and modified-Wisconsin method) in cattle, using only a level of contamination of 200 eggs per gram (EPG) of faeces and using GIN eggs collected from goat faeces to spike faecal samples from cattle. To further study the recovery rate of GIN eggs from cattle faeces, this study was conducted in two laboratories, one in Belgium and one in Italy to evaluate the sensitivity, accuracy, precision and reproducibility of the Mini-FLOTAC and McMaster techniques (at two reading levels: grids and chambers) for the detection of GIN eggs in spiked bovine faecal samples. METHODS: In both countries, spiked cattle faecal samples with five different levels of egg contamination (10, 50, 100, 200 and 500 EPG) of GINs were used. The study was performed in both laboratories by the same expert operator and using the same standard operating procedures (SOPs) for the Mini-FLOTAC and McMaster techniques. Sensitivity, accuracy and precision were calculated for each technique and for each level of contamination. Statistical analyses were performed to evaluate differences in performance between the two techniques. RESULTS: Mini-FLOTAC had a higher sensitivity (100% at all EPG levels for Mini-FLOTAC vs 0-66.6% for McMaster chambers and grids at levels< 100 EPG), a higher accuracy (98.1% mean value for Mini-FLOTAC vs 83.2% for McMaster grids and 63.8% for McMaster chambers) and a lower coefficient of variation (10.0% for Mini-FLOTAC vs 47.5% for McMaster grids and 69.4% for McMaster chambers) than McMaster. There was no significant difference in the recovery of GIN eggs between the two studies performed in Belgium and in Italy. CONCLUSIONS: The high GIN egg recovery rate detected by Mini-FLOTAC and the similar results obtained in Belgium and in Italy indicated that the diagnostic performance of a FEC technique was not dependent on the laboratory environment.

Cryptosporidium and Giardia in surface water and drinking water: Animal sources and towards the use of a machine-learning approach as a tool for predicting contamination.

Cryptosporidium and Giardia are important parasites due to their zoonotic potential and impact on human health, often causing waterborne outbreaks of disease. Detection of (oo)cysts in water matrices is challenging and few countries have legislated water monitoring for their presence. The aim of this study was to investigate the presence and origin of these parasites in different water sources in Northern Greece and identify interactions between biotic/abiotic factors in order to develop risk-assessment models. During a 2-year period, using a longitudinal, repeated sampling approach, 12 locations in 4 rivers, irrigation canals, and a water production company, were monitored for Cryptosporidium and Giardia, using standard methods. Furthermore, 254 faecal samples from animals were collected from 15 cattle and 12 sheep farms located near the water sampling points and screened for both parasites, in order to estimate their potential contribution to water contamination. River water samples were frequently contaminated with Cryptosporidium (47.1%) and Giardia (66.2%), with higher contamination rates during winter and spring. During a 5-month period, (oo)cysts were detected in drinking-water (<1/litre). Animals on all farms were infected by both parasites, with 16.7% of calves and 17.2% of lambs excreting Cryptosporidium oocysts and 41.3% of calves and 43.1% of lambs excreting Giardia cysts. The most prevalent species identified in both water and animal samples were C. parvum and G. duodenalis assemblage AII. The presence of G. duodenalis assemblage AII in drinking water and C. parvum IIaA15G2R1 in surface water highlights the potential risk of waterborne infection. No correlation was found between (oo)cyst counts and faecal-indicator bacteria. Machine-learning models that can predict contamination intensity with Cryptosporidium (75% accuracy) and Giardia (69% accuracy), combining biological, physicochemical and meteorological factors, were developed. Although these prediction accuracies may be insufficient for public health purposes, they could be useful for augmenting and informing risk-based sampling plans.

Anthelmintic resistance and common worm control practices in sheep farms in Flanders, Belgium.

In contrast to many other European countries, no data were available on the presence of anthelmintic resistance in gastrointestinal nematodes in sheep in Belgium. A faecal egg count reduction test was performed in 26 sheep flocks in Flanders, Northern Belgium. Results indicated widespread resistance against benzimidazoles (albendazole, fenbendazole and mebendazole), with treatment failure on all 8 farms investigated. Haemonchus contortus and Teladorsagia circumcincta were the predominant species after treatment failure. Amino acid substitutions associated with benzimidazole resistance were detected at the codon positions 167 (8%) and 200 (92%) of the isotype-1 beta tubulin gene in H. contortus, codon positions 198 (47%) and 200 (43%) in T. circumcincta and position 200 (100%) in T. colubriformis. Resistance against macrocyclic lactones (ivermectin, doramectin and moxidectin) was recorded on 7 out of 20 flocks, mainly in H. contortus and T. circumcincta. Treatment failure was also observed for closantel (in combination with mebendazole) and for monepantel, on one farm each. Trichostrongylus spp. were implicated with resistance against monepantel. A questionnaire survey on farm management and worm control measures indicated that worm control was often not sustainable. Ewes and lambs were treated frequently (on average 2.6 and 3.2 times per year), mostly without weighing. Only few sheep farmers (9%) regularly used faecal egg counts to monitor worm infections. Despite the FECRT showing otherwise, most of the farmers perceived the efficacy of anthelmintics as very good (30%) or good (54%).

Resistance against macrocyclic lactones in Psoroptes ovis in cattle.

BACKGROUND: Psoroptic mange is an important disease in Belgian Blue cattle. Treatment failure of macrocyclic lactones against Psoroptes ovis has been reported, but clear evidence of in vivo resistance is lacking. This study assessed the efficacy of macrocyclic lactone products on 16 beef farms in Belgium and the Netherlands in vivo and in vitro. METHODS: On each farm a group of animals (n = 7-14) with psoroptic mange was treated with two subcutaneous injections of a macrocyclic lactone product with 7-10 days interval (15 farms) or a single injection with a long-acting macrocyclic lactone (1 farm). In vivo efficacy was assessed by the reduction in mite counts, clinical index (proportion of the body surface affected by lesions), the proportion of the animals with negative mite counts after the first treatment round and the number of treatment rounds needed to obtain zero mites counts in all animals. A mite population was categorized as sensitive when the mite count reduction after the first treatment round > 95% and the lower limit of the uncertainty interval > 90%. Resistance was detected when both parameters were below their threshold and suspected when one parameter was too low. In vitro knockdown and mortality were evaluated in a contact test. RESULTS: The proportion of the animals with negative mite counts after the first treatment round varied from 0 to 80%. All farms needed two or more treatments rounds to obtain zero mite counts on all animals. Clinical index only started to reduce after the second treatment round. Mite populations from three farms were categorized as sensitive, one as suspected resistant and 12 as resistant. No correlation was found between in vitro lethal dose 50 and knockdown dose 50 values and in vivo efficacy parameters. CONCLUSIONS: Unambiguous treatment failure was detected on 12 out of 16 farms, confirming the presence of macrocyclic lactone resistance on Belgian Blue beef farms. In vitro parameters could not discriminate the farms based on their in vivo sensitivity. The mean reduction in mite counts and the lower limit of the confidence interval are proposed as parameters to identify acaricide resistance.