High rates of benzimidazole-resistance-associated alleles in Haemonchus contortus and detection of resistance against macrocyclic lactones in strongylids from German alpaca herds.

The population of South American camelids (SAC) has been steadily growing in Europe, where they are confronted with the regional endoparasite population of ruminants. As there are no anthelmintic drugs registered for use against nematode infections in SACs, anthelmintics (AH) available for ruminants or horses are usually applied. Reports indicating potential failures in administered AH are increasing. However, the generally low egg counts in SACs complicate the application of resistance tests in the field. The present study reports a follow-up study on SAC farms where anthelmintic resistance (AR) was suspected. The aims were (i) to repeat faecal egg count reduction tests (FECRTs) on potentially affected farms identified in a previous study with larger sample sizes, (ii) to verify suspected AR of Haemonchus contortus against benzimidazoles (BZ) by performing a single-nucleotide polymorphism (SNP) analysis using digital polymerase chain reaction (dPCR), and (iii) to apply the mini-FLOTAC technique for more reliable results at low egg counts in line with current recommendations. Seven farms (9-46 animals each) were examined by coproscopy, larval differentiation and SNP analysis. A FECRT was performed on six of these farms with moxidectin (three farms), monepantel (two farms) and ivermectin (one farm). The FEC was calculated according to the current World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines with the clinical protocol (a newly introduced variant of FECRT which can be used for smaller sample sizes and lower egg counts on the cost of sensitivity) and an expected efficacy of 99%. A high level (> 90%) of BZ-resistance-associated SNPs on codon 200 of H. contortus was observed on all farms. With the FECRT, resistance was demonstrated for ivermectin (74% FECR), while it remained inconclusive for one farm for moxidectin treatment. Sustained efficacy was demonstrated for the remaining treatments. This study showed an advanced level of BZ resistance in H. contortus of SACs and the development of AR against macrocyclic lactones on some farms. Thus, constant monitoring of AH treatment and sustainable worm control methods both need to be applied.

Correlation of Faecal Egg Counts with Clinical Parameters and Agreement between Different Raters Assessing FAMACHA©, BCS and Dag Score in Austrian Dairy Sheep.

Gastrointestinal nematodes, most notably trichostrongylids, are known to cause significant losses in sheep production. Previous studies have shown that monitoring parameters (e.g., FAMACHA©, BCS, dag score) change with increasing egg excretion. These parameters are well known and frequently used for targeted selective treatment. Based on the willingness to participate in this study (based on a previous questionnaire distribution among sheep farmers in Austria) we investigated the associations between faecal egg counts and the FAMACHA©, BCS, and dag scores of 1195 dairy ewes. Faecal samples were analysed using the Mini-FLOTAC technique I and larval culture. Three raters assessed the FAMACHA©, BCS, and dag scores in sheep to calculate the inter-rater agreement and intraclass correlation coefficient. The responses to the questionnaire of 23 farms were used for the evaluation, of which 16 farms were visited. Trichostrongylid eggs were detected in 95% of the faecal samples. The BCS was negatively correlated with the eggs per gram of faeces (EpG) (r = -0.156; p < 0.001) and the FAMACHA© score was slightly positively correlated with EpG (r = 0.196; p < 0.001). A small proportion of sheep (25%) shed the majority of eggs (47% to 84%). A moderate to good agreement for the parameters was found between the raters. In conclusion, the clinical parameters showed only weak correlations with faecal egg counts, and we confirmed that a minority of the flock is responsible for the majority of the pasture contamination with trichostrongylid eggs. Clinical raters should be trained before a study to increase the agreement between them.

Parasites and zoonotic bacteria in the feces of cats and dogs from animal shelters in Carinthia, Austria.

Due to their close associations with humans, dogs and cats can be important reservoirs for zoonotic pathogens. In the current study 200 fecal samples of dogs (n = 70 samples) and cats (n = 130 samples) from animal shelters in Carinthia, southern Austria, were examined for the presence of parasites (fecal flotation and larval migration assay) and selected bacteria. Overall, 17.1% of the canine and 38.5% of the feline samples were positive for parasites (p < 0.001), most commonly Giardia duodenalis (dogs and cats), including potentially zoonotic genotypes revealed by multilocus genotyping, and Toxocara cati (cats). Cryptosporidium (C. felis), Cystoisospora spp. (dogs and cats), hookworms (dog), Trichuris (dog) Capillaria hepatica (cats), taeniids (cat), and Aelurostrongylus abstrusus (cat) were also found. Zoonotic bacteria were detected in 10.5% of the samples, Salmonella enterica (dogs), Campylobacter jejuni (dogs and cats) and Yersinia enterocolitica (cat) and were significantly associated with parasite infections in cats but not in dogs. Samples that were positive for several pathogens were common; especially G. duodenalis and T. cati were frequently found in association with each other, other parasites or bacteria. The spectrum of detected pathogens is comparable to that of other dog and cat populations in central Europe. However, since animals from shelters are frequently rehomed, diagnostic measures, appropriate hygiene and therapy as well as training of shelter staff are recommended to prevent zoonotic transmission of enteropathogens to staff or new owners. The presence of heteroxenic parasites, i.e. Aelurostrongylus abstrusus and Taenia taeniaeformis, and spurious excretion of Ca. hepatica in cats, indicates that these animals preyed on intermediate hosts, and that biosafety measures in pet shelters need to be evaluated for their efficacy in the prevention of pathogen transmission.

Presence of Equine and Bovine Coronaviruses, Endoparasites, and Bacteria in Fecal Samples of Horses with Colic.

Acute abdominal pain (colic) is one of the major equine health threats worldwide and often necessitates intensive veterinary medical care and surgical intervention. Equine coronavirus (ECoV) infections can cause colic in horses but are rarely considered as a differential diagnosis. To determine the frequency of otherwise undetected ECoV infections in horses with acute colic, fresh fecal samples of 105 horses with acute colic and 36 healthy control horses were screened for viruses belonging to the Betacoronavirus 1 species by RT-PCR as well as for gastrointestinal helminths and bacteria commonly associated with colic. Horses with colic excreted significantly fewer strongyle eggs than horses without colic. The prevalence of anaerobic, spore-forming, gram-positive bacteria (Clostridium perfringens and Clostridioides difficile) was significantly higher in the feces of horses with colic. Six horses with colic (5.7%) and one horse from the control group (2.8%) tested positive for Betacoronaviruses. Coronavirus-positive samples were sequenced to classify the virus by molecular phylogeny (N gene). Interestingly, in three out of six coronavirus-positive horses with colic, sequences closely related to bovine coronaviruses (BCoV) were found. The pathogenic potential of BCoV in horses remains unclear and warrants further investigation.

Development of a three-colour digital PCR for early and quantitative detection of benzimidazole resistance-associated single nucleotide polymorphisms in Haemonchus contortus.

Haemonchus contortus is the most pathogenic nematode in small ruminants and anthelmintic resistance (AR) hampers its efficient control. Early detection of AR status is required to reduce selection for AR and cannot be achieved using phenotypic tests. For benzimidazoles (BZs), the detection of AR-associated alleles characterised by single nucleotide polymorphisms (SNPs) in the isotype 1 ß-tubulin gene allows early AR detection in strongyles. The F200Y, F167Y, E198A and E198L polymorphisms have been described in BZ-resistant populations with a clear variation in frequencies between regions. A novel digital PCR (dPCR) enables the detection of all of the above-described polymorphisms in H. contortus. Assays were validated using synthetic DNA fragments containing these SNPs. Then, larvae obtained and pooled at farm level from 26 Austrian and 10 Italian sheep farms were analysed. For all assays a detection limit of 15 copies/µl of resistance alleles and a high level of accuracy were demonstrated, allowing to detect allele frequencies of 1% in most samples. In Austrian samples, elevated frequencies of F200Y resistance alleles were detected on all farms. Polymorphisms in codon 167 and codon 198 were identified in H. contortus from Austria for the first time. In Italian samples, the frequency of resistance alleles was still comparatively low, but F200Y resistance alleles were traceable. In conclusion we developed for the first time dPCR assays that target all SNPs of relevance associated with BZ-resistance in H. contortus. Future research on AR development could benefit from an early onset of SNP-based surveillance that would include the developed assays for all SNPs of relevance. Improved surveillance in the long term will include other important, though less pathogenic, nematode genera in the analyses.

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.

Enterocytozoon bieneusi in fecal samples from calves and cows in Austria.

Enterocytozoon bieneusi is an obligate intracellular pathogen that infects livestock, companion animals, and wildlife and has the potential to cause severe diarrhea especially in immunocompromised humans. In the underlying study, fecal samples from 177 calves with diarrhea and 174 adult cows originating from 70 and 18 farms, respectively, in Austria were examined for the presence of E. bieneusi by polymerase chain reaction targeting the Internal Transcribed Spacer 1 (ITS1) region. All positive samples were further sequenced for genotype determination. Overall, sixteen of the 351 (4.6%) samples were positive for E. bieneusi, two of the 174 samples from cows (1.2%) and 14 of the 177 samples from calves (7.9%). In total, four genotypes, J (n = 2), I (n = 12), BEB4 (n = 3), and BEB8 (n = 1), were identified. The uncorrected p-distance between the four ITS1 lineages (344 bp) ranges from 0.3% to 2.9%. The lineages differ by 1 bp (I and J), 2 bp (J and BEB4), and 3 bp (I and BEB4), respectively, and BEB8 differs by 7 to 10 bp from the latter three lineages. Two of the E. bieneusi-positive calves showed an infection with two different genotypes. E. bieneusi occurred significantly more often in calves > 3 weeks (8/59) than in calves ≤ 3 weeks (6/118), respectively (p = 0.049). Calves with a known history of antimicrobial treatment (50 of 177 calves) shed E. bieneusi significantly more often than untreated calves (p = 0.012). There was no statistically significant difference in E. bieneusi shedding in calves with or without a medical history of antiparasitic treatment (p = 0.881). Calves showing a co-infection with Eimeria spp. shed E. bieneusi significantly more often than uninfected calves (p = 0.003). To our knowledge, this is the first report of E. bieneusi in cattle in Austria. Cattle should be considered as a reservoir for human infection since potentially zoonotic E. bieneusi genotypes were detected.

Haemonchosis in Sheep and Goats, Control Strategies and Development of Vaccines against Haemonchus contortus.

The evolutionary success of parasitic worms causes significant economic losses and animal health problems, including in the small ruminant industry. The hematophagous nematode Haemonchus contortus is a common endoparasite that infects wild and domestic ruminants worldwide, especially in tropical and subtropical regions. To date, the most commonly applied control strategy is the administration of anthelminthic drugs. The main disadvantages of these chemicals are their ecotoxic effects, the necessary withdrawal period (especially important in dairy animals) and the increasing development of resistance. Vaccines offer an attractive alternative control strategy against Haemonchus infections. In previous years, several potential vaccine antigens prepared from H. contortus using the latest technologies have been assessed in clinical trials using different methods and strategies. This review highlights the current state of knowledge on anti-H. contortus vaccines (covering native, recombinant and DNA-based vaccines), including an evaluation, as well a discussion of the challenges and achievements in developing protective, efficient, and long-lasting vaccines to control H. contortus infection and haemonchosis in small ruminants. This paper also addresses novel developments tackling the challenge of glycosylation of putative candidates in recombinant form.

Trichurosis on a Conventional Swine Fattening Farm with Extensive Husbandry-A Case Report.

Helminth infections of swine regain clinical and economic importance due to the increasing demand for pork from extensive husbandry. Infections with Trichuris suis in pigs can lead to wasting and diarrhoea. This was demonstrated by a case of clinical trichurosis on a conventional fattening farm, where pigs were kept on pasture. While all pre-fattening pigs, which had not been on the pasture yet, had a good body condition and firm faeces, diarrhoea and poor body condition were observed in approximately half of the fattening pigs kept on pasture. Rectally collected faecal samples from all animals were investigated using faecal flotation. High numbers of T. suis eggs were detected in 17 out of 32 faecal samples, while all samples from pre-fattening pigs were negative. The highest number of eggs per gram of faeces was 778,000. Two out of three environmental samples were also positive for T. suis in faecal flotation. This case demonstrates that T. suis must be considered as an enteropathogen in pigs kept on pasture, as favourable environmental conditions, and the lack of removal of faeces from a pasture can lead to the accumulation of large numbers of infective eggs in the pigs' surroundings.

Eprinomectin and Moxidectin Resistance of Trichostrongyloids on a Goat Farm in Austria.

Anthelmintic resistance (AR) of trichostrongyloids is widespread in Europe, but there is no up-to-date information on the spread of AR in caprine parasites in Austria. Eprinomectin (EPR) is currently the only anthelmintic drug of the macrocyclic lactones registered for goats in Europe. The aim of the present study was to gather information regarding the efficacy of anthelmintics against trichostrongyloids on a dairy goat farm in Austria with reported treatment failure of macrocyclic lactones and to determine the presence of different trichostrongyloid genera. Faecal egg count reduction tests (FECRT) using Mini-FLOTAC were performed with eprinomectin (EPR) and moxidectin (MOX). Egg count reduction, calculated with the R package egg-Counts, was 44% for EPR and 86% for MOX, confirming AR of trichostrongyloids for both compounds. The most frequently detected genus in larval cultures was Haemonchus, followed by Trichostrongylus. This is the first report of MOX resistance in caprine trichostrongyloids in Europe. Failure of EPR and MOX to control trichostrongyloid infections is a severe threat to dairy goat farming, since other compounds must not be applied in goats used for milk production. Haemonchus contortus is one of the most pathogenic parasites of small ruminants and can quickly develop AR. Thus, immediate action should be taken to slow the further spread of AR in this and other roundworm species of ruminants in Austria.

Strongyloides stercoralis infection in dogs in Austria: two case reports.

BACKGROUND: Strongyloides stercoralis is endemic in tropical and subtropical regions, but reports of infections in central and northern Europe have been recently increasing. Infections occur mainly in humans and dogs. In dogs, both dog-adapted and zoonotic S. stercoralis genotypes seem to occur. Clinical manifestations mainly include gastrointestinal and respiratory signs. The severity of the disease can vary greatly and depends on the immune status of the host. The infection is potentially fatal in immunosuppressed individuals, either medically induced or due to an underlying disease, in which hyperinfections and disseminated infections with extraintestinal parasite dissemination may occur. METHODS: Diagnosis was based on coproscopy, including flotation and the Baermann funnel technique, histology of small intestinal biopsies and molecular analysis of mitochondrial cytochrome oxidase subunit I (COI) and hypervariable regions I and IV (HVR I and HVR IV) of the nuclear 18S rDNA loci. RESULTS: Two independent cases of severe canine S. stercoralis infection in Austria are presented. In both cases, S. stercoralis was detected in histological sections of the small intestine and with the Baermann funnel technique. Molecular analysis revealed strains with zoonotic potential. Case 1 was a 1-year-old female French bulldog with a long history of respiratory and gastrointestinal signs, severe emaciation and apathy before S. stercoralis infection was diagnosed. Treatment with moxidectin (2.5 mg/kg body weight [BW], oral route) did not eliminate the infection, but treatment with ivermectin (0.2 mg/kg BW, subcutaneously) was successful. Case 2 consisted of two 2-month-old Pomeranian puppies, one female and one male, from a litter of four, which died soon after presenting dyspnoea and haemorrhagic diarrhoea (female) or torticollis (male); S. stercoralis infection was first diagnosed post-mortem. CONCLUSION: More attention should be paid to this nematode because although it appears to be rare in Austria, it is easily overlooked on standard coproscopy unless a Baermann funnel technique is used, and even then, it can be missed. Moxidectin is not always successful in eliminating the infection, and treatment with ivermectin should be considered in cases of infection.

Serological and faecal detection of Angiostrongylus vasorum in dogs from Austria.

Canine angiostrongylosis is a potentially lethal parasitic disease that can manifest itself with a broad spectrum of clinical signs, including respiratory distress, neurological and bleeding disorders, or non-specific signs. The occurrence of Angiostrongylus vasorum is widely reported in Europe, but very little is known about its presence in Austria. In this first large-scale survey, 1279 sera were collected from Austrian dogs and tested by an ELISA for the detection of circulating antigen of A. vasorum (sensitivity: 95.7%, specificity 94.0%) and by a separate ELISA detecting specific antibodies (sensitivity 81.0%, specificity 98.8%). Furthermore, 1040 faecal samples were tested for the presence of lungworm first stage larvae (L1). One dog (0.1%, 95% confidence intervals [CI]: 0.0-0.4%) was positive in both ELISAs, while 1.2% (n = 15, CI: 0.7-1.9%) of the tested dogs were antigen-positive and 1.5% (n = 19, CI: 0.9-2.3%) were positive for specific antibodies. Overall, 13 dogs (1.3%; CI: 0.7-2.1%) were positive for A. vasorum L1 while 31 dogs were positive for Crenosoma vulpis L1 (3.0%; CI: 2.0-4.2%). One dog shed L1 from both A. vasorum and C. vulpis (0.1%, CI: 0.0-0.5%). Dogs positive for A. vasorum originated from northeast, southeast and south Austria (antigen and/or antibody detection), but also from north, west and southwest Austria (antibody detection) and from northeast and west Austria (L1 detection). One of 88 blood samples (1.1%, CI: 0.0-6.2%) submitted from the eastern part of Austria was positive by a rapid assay for A. vasorum antigen detection (Angio Detect™). Crenosoma vulpis positive samples originated from northwest, north, northeast, south and west Austria. These results confirm the very sporadic occurrence of A. vasorum in the investigated areas of the country. However, due to the substantial infectious pressure from the surrounding countries and the free circulation of dogs and foxes acting as wildlife reservoirs and due to clinical relevance for infected dogs, it is crucial to maintain disease awareness also in areas where the parasite has not yet been detected.

Co-infection with Babesia canis and Babesia gibsoni in a dog.

A four-year-old intact male Boxer, that had a history of travelling to Serbia, was referred for lethargy and anaemia. Shortly before the dog was referred, it was diagnosed twice with an infection with Babesia canis and was treated with imidocarb both times. A blood smear evaluation was indicative of the presence of intraerythrocytic piroplasms. After receiving inconclusive results regarding the type of piroplasm, the dog was diagnosed with simultaneous infections with B. canis and Babesia gibsoni via real-time polymerase chain reaction (rt-PCR) testing. The dog was treated with imidocarb, atovaquone and azithromycin, and in a follow-up examination, the PCR results were negative for B. canis and B. gibsoni. Several weeks later, the dog was presented again, and a PCR was positive for B. gibsoni. After atovaquone and azithromycin failed to eliminate the parasites, a therapy attempt using metronidazole, clindamycin and doxycycline was initiated. Six months after diagnosis, the treatment appeared successful in eliminating B. gibsoni. This case report describes the clinical findings of the co-infection and the initiated diagnostic and therapeutic approaches.

DNA barcoding of rumen flukes (Paramphistomidae) from bovines in Germany and Austria.

Rumen flukes have received growing veterinary attention in western and central Europe during the past two decades because of an increase in prevalence of infection in cattle and sheep, including cases of severe clinical disease. Historically, rumen fluke infections in Europe were assumed to be caused mainly by Paramphistomum cervi (or species, which were later considered to be synonymous with P. cervi), but more recently molecular studies demonstrated Calicophoron daubneyi to be the predominating species. For the present investigation, adult rumen flukes isolated from 23 cattle originating from ten farms in Germany (Saxony [1], Baden-Württemberg [4], Bavaria [5]) and one farm in Austria (Tyrol) were analyzed to establish partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) and the complete sequence of the nuclear internal transcribed spacer 2 (ITS2). Flukes of five animals (dairy cows from three farms in Bavaria) were determined as P. leydeni, and flukes of 18 animals (dairy cows or cattle from cow-calf operations from eight farms in Saxony [1], Baden-Württemberg [4], Bavaria [2], and Tyrol [1]) were identified as C. daubneyi. Based on the molecular analysis of adult rumen flukes collected from cattle, the results of this investigation confirm the common occurrence of C. daubneyi in Germany and reveal the first definitive findings of P. leydeni in Germany and C. daubneyi in Austria.

Absorption and Distribution of Toltrazuril and Toltrazuril Sulfone in Plasma, Intestinal Tissues and Content of Piglets after Oral or Intramuscular Administration.

Piglet coccidiosis due to Cystoisospora suis is a major cause of diarrhea and poor growth worldwide. It can effectively be controlled by application of toltrazuril (TZ), and oral formulations have been licensed for many years. Recently, the first parenteral formulation containing TZ in combination with iron (gleptoferron) was registered in the EU for the prevention of coccidiosis and iron deficiency anemia, conditions in suckling piglets requiring routine preventive measures. This study evaluated the absorption and distribution of TZ and its main metabolite, toltrazuril sulfone (TZ-SO2), in blood and intestinal tissues after single oral (20 mg/kg) or single intramuscular (45 mg/piglet) application of TZ. Fifty-six piglets were randomly allocated to the two treatment groups. Animals were sacrificed 1-, 5-, 13-, and 24-days post-treatment and TZ and TZ-SO2 levels were determined in blood, jejunal tissue, ileal tissue, and mixed jejunal and ileal content (IC) by high performance liquid chromatography (HPLC). Intramuscular application resulted in significantly higher and more sustained concentrations of both compounds in plasma, intestinal tissue, and IC. Higher concentrations after oral dosing were only observed one day after application of TZ in jejunum and IC. Toltrazuril was quickly metabolized to TZ-SO2 with maximum concentrations on day 13 for both applications. Remarkably, TZ and TZ-SO2 accumulated in the jejunum, the primary predilection site of C. suis, independently of the administration route, which is key to their antiparasitic effect.

Multispecific resistance of sheep trichostrongylids in Austria.

Anthelmintic overuse and failure to implement methods preventing the development and spread of anthelmintic resistance (AR) have led to an alarming increase of resistant ovine trichostrongylids worldwide. The aim of the present study was to determine whether the routine anthelmintic treatment strategy was effective, to obtain insights into the frequency of AR in trichostrongylids of sheep in Austria, and to determine the presence of different trichostrongylid genera. On 30 sheep farms, the faecal egg count reduction test (FECRT) was performed with the Mini-FLOTAC technique in two consecutive studies. In study 1, only fenbendazole and moxidectin were tested, while different compounds and products were used in study 2. Overall, 33 treatment groups were formed: 11 groups were treated with benzimidazoles (fenbendazole and albendazole), 2 groups with avermectins (ivermectin, doramectin), 18 groups with moxidectin, and two groups with monepantel. Reduced efficacy was detected in 64%, 100%, 28% and 50% of these groups, respectively. The most frequently detected genus in larval cultures was Haemonchus, which had been barely detected in Austria previously, followed by Trichostrongylus. Multispecific resistance of trichostrongylids in Austria seems to be on the rise and H. contortus was detected unexpectedly frequently in comparison to previous studies. There is an urgent need to develop efficient communication strategies aimed at improving the engagement of farmers and veterinarians in sustainable parasite control.

TITLE: Résistance multispécifique des trichostrongylidés des ovins en Autriche. ABSTRACT: La surutilisation des anthelminthiques et l'échec de la mise en œuvre de méthodes empêchant le développement et la propagation de la résistance aux anthelminthiques (RA) ont conduit à une augmentation alarmante des trichostrongylidés ovins résistants dans le monde. Le but de nos études était de déterminer si la stratégie de traitement anthelminthique de routine était efficace, d'avoir un aperçu de la fréquence de la RA chez les trichostrongylidés des moutons en Autriche et de déterminer la présence de différents genres de trichostrongylidés. Dans 30 élevages ovins, le test de réduction du nombre d'œufs fécaux (FECRT) a été réalisé avec la technique Mini-FLOTAC dans deux études consécutives. Dans l'étude 1, seuls le fenbendazole et la moxidectine ont été testés, tandis que différents composés et produits ont été utilisés dans l'étude 2. Au total, trente-trois groupes de traitement ont été formés, 11 groupes ont été traités avec des benzimidazoles (fenbendazole et albendazole), 2 groupes avec des avermectines (ivermectine, doramectine), 18 groupes avec la moxidectine et deux groupes avec le monepantel. Une efficacité réduite a été détectée dans 64 %, 100 %, 28 % et 50 % de ces groupes, respectivement. Le genre le plus fréquemment détecté dans les cultures larvaires était Haemonchus, qui avait été rarement détecté en Autriche auparavant, suivi de Trichostrongylus. La résistance multispécifique des trichostrongylidés en Autriche semble augmenter et H. contortus a été détecté fréquemment, de manière inattendue par rapport aux études précédentes. Il est urgent de développer des stratégies de communication efficaces visant à améliorer l'engagement des éleveurs et des vétérinaires dans le contrôle durable des parasites.

Piglet coccidiosis in Belgium and the Netherlands: Prevalence, management and potential risk factors.

Piglet coccidiosis caused by Cystoisospora suis is one of the most important causes of diarrhea in suckling piglets. The parasite has a fast development and multiplies quickly and effectively under the conditions of a farrowing unit. Control measures include cleaning and disinfection and anticoccidial treatment. In Europe, toltrazuril-based products are authorized for this purpose and are applied to piglets on affected farms in the first week of life. To observe the effect of treatment and disinfection on the control of piglet coccidiosis in the field, 23 farms (11 from Belgium, 12 from the Netherlands, mean number of sows = 1413) were sampled twice by litter in the second and third week of life and fecal scores and the presence of C. suis oocysts were determined. A questionnaire was used to collect data on farm sizes, management and hygiene measures as well as treatment (product, dose and piglet age). Thirteen farms regularly treated with toltrazuril (treatment age: 1-6 days, mean 4.3 days) and 19 applied disinfection. Parasite excretion was documented on 60.9% and diarrhea on 78.3% of farms and in 34.3%/15.7% of the litters. Only 2.4% of the litters showed both, so subclinical infection appeared to be common. No significant differences between farms that did not treat against coccidiosis and farms that applied toltrazuril was observed with regard to C. suis oocysts shedding and/or diarrhea. However, in litters that were treated within the first three days of life, oocyst excretion was significantly less often observed than in litters with later treatment (p = 0.033). No significant effect of disinfection could be shown, but most farms applied disinfectants that have no proven effect against coccidia (oxygen-releasing agents or glutaraldehyde + ammonia) while the only farm that used chlorocresols (which are effective against coccidia) did not show oocyst shedding. Current control measures thus appear to be insufficient on most of the examined farms. It is therefore recommended to treat piglets timely and to apply effective disinfectants to reduce C. suis infections. Furthermore, regular evaluation of sustained efficacy of all implemented measures are necessary.

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.

Cryptosporidium parvum, Cryptosporidium ryanae, and Cryptosporidium bovis in samples from calves in Austria.

Fecal samples of 177 calves of up to 180 days of age with diarrhea from 70 farms in Austria were examined to obtain information on the occurrence of Cryptosporidium species. Initially, all samples were examined by phase-contrast microscopy. Cryptosporidium-positive samples (55.4%; n = 98) were screened by gp60 PCR, resulting in 68.4% (n = 67) C. parvum-positive samples. The remaining 31 gp60-PCR-negative and the phase-contrast microscopy negative samples (n = 79) were screened by PCR targeting a 700 bp fragment of the 18S rRNA gene. Sequencing of the PCR products revealed the presence of C. parvum (n = 69), C. ryanae (n = 11), and C. bovis (n = 7). The latter two species have never been described in Austria. C. parvum-positive samples were genotyped at the gp60 gene locus, featuring four subtypes (IIaA15G2R1, IIaA21G2R1, IIaA19G2R1, IIaA14G1R1). The most frequently detected subtype IIaA15G2R1 (n = 52) was present in calves from 30 different farms. IIaA14G1R1 (n = 5) occurred on a single farm, subtype IIaA21G2R1 (n = 4) on two farms, and subtype IIaA19G2R1 (n = 4) on three farms. The results confirm the widespread occurrence of zoonotic C. parvum in diarrheic calves.