Farmers' and Experts' Knowledge Coping with Sheep Health, Control and Anthelmintic Resistance of Their Gastrointestinal Nematodes.

Gastrointestinal nematodes are common in grazing sheep, but the intensity of the infection is not easily appreciated by farmers. For decades, they have relied on anthelmintic treatments to control these gastrointestinal nematodes. This has led to anthelmintic resistance and poor control of infection in most regions of the world. Using face-to-face semidirective interviews with farmers, we recorded their views on gastrointestinal nematode infection and its control. Ten organic and nine conventional meat sheep farmers from central France and 20 milk sheep farmers from the Basque region were interviewed and the data were analysed using a health model based on the importance of the disease and the barriers to implementing control. It was found that gastrointestinal nematodes were not the main health concern, and therefore farmers were not willing to invest too much time and money in their control. The conventional farmers relied on their veterinarians (the experts) to organise and select the anthelmintics, although they complained about the limited investment of their veterinarians in this matter. Organic farmers complained about their lack of knowledge about complementary and alternative medicines. Farmers rarely used faecal egg counts to build control of gastrointestinal nematodes and were unaware of the intensity of their infection. Knowledge of anthelmintic resistance was general (it exists) but farmers did not know if it existed on their farm. Resistance was often considered to have come from elsewhere, so the farmer did not feel at fault and did not take measures to prevent resistance. There is a need for all stakeholders to use faecal egg counts to assess the intensity of infection as well as the level of anthelmintic resistance to establish individual farm control programmes rather than standard blanket treatments.

Haemonchus contortus Adopt Isolate-Specific Life History Strategies to Optimize Fitness and Overcome Obstacles in Their Environment: Experimental Evidence.

Gastrointestinal nematodes (GIN) use flexible life history strategies to maintain their fitness under environmental challenges. Costs incurred by a challenge to one life trait can be recouped by increasing the expression of subsequent life traits throughout their life cycle. Anticipating how parasites respond to the challenge of control interventions is critical for the long-term sustainability of the practice and to further ensure that the parasites withstand favourable adaptive responses. There is currently limited information on whether distinct populations of a GIN species respond to the same environmental challenge in a consistent manner, with similar alterations to their life history strategies or comparable fitness outcomes. This study compared the life history traits and experimental fitness of three distinct Haemonchus contortus isolates exposed to environmental challenges at both the parasitic (i.e., passage through resistant or susceptible sheep) and free-living (i.e., exposure to diverse climatic conditions) life stages. The key findings show that H. contortus maintain their fitness under challenge with isolate-specific alterations to their life history strategies. Further, partial exploration of the H. contortus isolates transcriptomes using cDNA-AFLP methods confirmed disparate expression profiles between them. These results bring fresh insights into our understanding of the non-genetic adaptive processes of GIN that may hinder the efficacy of parasite control strategies.

Anthelmintic Treatment and the Stability of Parasite Distribution in Ruminants.

Parasites are generally overdispersed among their hosts, with far-reaching implications for their population dynamics and control. The factors determining parasite overdispersion have long been debated. In particular, stochastic parasite acquisition and individual host variation in density-dependent regulation through acquired host immunity have been identified as key factors, but their relative roles and possible interactions have seen little empirical exploration in parasite populations. Here, Taylor's power law is applied to test the hypothesis that periodic parasite removal destabilises the host-parasite relationship and increases variance in parasite burden around the mean. The slope of the power relationship was compared by analysis of covariance among 325 nematode populations in wild and domestic ruminants, exploiting that domestic ruminants are often routinely treated against parasite infections. In Haemonchus spp. and Trichostrongylus axei in domestic livestock, the slope increased with the frequency of anthelmintic treatment, supporting this hypothesis. In Nematodirus spp., against which acquired immunity is known to be strong, the slope was significantly greater in post-mortem worm burden data than in faecal egg counts, while this relationship did not hold for the less immunogenic genus Marshallagia. Considered together, these findings suggest that immunity acting through an exposure-dependent reduction in parasite fecundity stabilises variance in faecal egg counts, reducing overdispersion, and that periodic anthelmintic treatment interferes with this process and increases overdispersion. The results have implications for the diagnosis and control of parasitic infections in domestic animals, which are complicated by overdispersion, and for our understanding of parasite distribution in free-living wildlife. Parasite-host systems, in which treatment and immunity effectively mimic metapopulation processes of patch extinction and density dependence, could also yield general insights into the spatio-temporal stability of animal distributions.

Farmers' Views and Tools Compared with Laboratory Evaluations of Parasites of Meat Goats in French West Indies.

Gastrointestinal nematodes (GINs) are a major health problem in tropical goat husbandry. The control of GIN has been nearly exclusively reliant on the use of anthelmintic treatments. Their wide use has provoked the appearance and diffusion of anthelmintic resistance. Therefore, there is a need to use anthelmintics only when they are really needed. This strategy of targeted selective treatment (TST) has been recommended. The selection of animals to be treated has been based either on the objective measures of GIN intensity (fecal nematode egg counts) performed in the laboratory or on indirect assessment such as anemia (FAMACHA©), diarrhea score or weight gains, particularly in sheep. The roughness of hair has also been proposed in goats. These indicators can be handled by the farmer. Their opinion on the importance of GINs, and the indicators that they are ready to accept and use have very rarely been studied. Goat for meat production is important in the French West Indies (especially in Guadeloupe) and GIN infection may significantly alter this production. Eighteen farmers participated in semi-directive interviews in order to appreciate their relation to goat GIN infection and the solutions they considered. Seventeen farms were investigated for fecal nematode egg counts, FAMACHA©, body score, and roughness of hair. The average infection by GINs was high (average fecal egg count 1562 and standard deviation 2028) with a wide range from one farm to another (from 0 to 25,000 eggs of GIN per gram of feces). The Haemonchus genera was predominant (54%), followed by Trichostrongylus (37%) and Oesophagostomum (9%). Young goats were less infected than adult goats since they were not yet grazing; males were more infected than females; and the Creole breed was more infected than the other breeds. Among the farming types, the professional ones were less infected compared with the traditional or mixed agriculture and husbandry farms. Those using targeted selective treatment did not have a significantly higher GIN infection than those treating the whole herd. Most of the characteristics were related and multivariate analysis could not match the intensity of GIN infection with any parameter. The frequency of anthelmintic treatments was negatively related to the use of body score, FAMACHA©, and hair roughness. The use of semi-directive interviews provided a wider understanding of the strategies and problems of farmers. The farmers valued their animals very much and diseases, in general, were a preoccupation, whereas parasites were not a major issue for traditional farmers. This is due to the important use of indicators and the belief in their value that gives comfort to the farmers that the parasites are being controlled. The extension services have well diffused the practice of indicators to the goat farmers of Guadeloupe, with some depending less on anthelmintics to control the gastrointestinal nematodes by using targeted selective treatments.

Serial passage in resistant sheep drives the infectivity and fitness of Teladorsagia circumcincta in susceptible lambs: Experimental evidence.

Gastrointestinal nematodes (GIN) of small ruminants have adapted their life history strategies to thrive in diverse and fluctuating environments. Environments which alter their expression of life traits may also drive changes in the infection or transmission dynamics, particularly if transferred to a foreign setting. This study aimed to explore how repeated exposure to a resistant sheep host environment would alter the life history traits and infection dynamics of Teladorsagia circumcincta when consequently infected in susceptible lambs. Following just three generations of passage in resistant sheep, T. circumcincta significantly increased their infectivity and fitness in susceptible lambs compared to a control population. This is the first evidence to indicate the resistant host environment can drive such rapid changes in the expression of GIN life traits, with potentially undesirable epidemiological outcomes.

Varying Degrees of Animal Reification by Stakeholders in Experimental Research.

The attitude towards animals in research depends on both the role of the stakeholder and their personal characteristics. Most studies on the subject have been carried out on stakeholders from biomedical research institutes with comparatively few sociological studies on stakeholders from agricultural research centers. Previous findings suggest that animal caretakers at agricultural research centers felt undervalued by the hierarchy, and that animal reification was present in the sector. This may indicate that a lack of consideration for the animal subjects correlates with an inadequate sensitivity towards humans. Since these findings were published twenty years ago, there has been an increasing emphasis on the importance and actions of ethics committees in research, animal welfare bodies, and public concern for animals, which may have impacted the current perspective. To better understand current degrees of animal reification amongst stakeholders of agricultural research, we conducted semi-directive interviews at a leading agricultural research institute in France (INRAE). The interviews targeted both animal caretakers and researchers who were involved in the study of infectious diseases in livestock, or the behavior of horses and quails. After having transcribed the recorded interviews into text, semi-automatized analyses were carried out to categorize them into distinct groups, from which the most characteristic words and sentences were extracted. Three groups of stakeholders were identified: (i) animal caretakers involved in invasive infectious disease research; (ii) animal caretakers involved in behavioral research; and (iii) researchers. The findings show that animal caretakers felt acknowledged by their hierarchy. It is possible the increased skill criteria for people recruited into this position over the years, combined with greater prospects for continuous learning and development in the profession, may have fostered a more respectful regard across the hierarchy. The animal caretakers clearly expressed that their primary objective was to successfully execute the research protocols and that the animals were viewed as prototypes for research, with which they could, on occasion, develop a bond with. The bond was more important for animal caretakers involved in behavioral studies than for those involved in the study of infectious diseases, where invasive biological sampling and restraining of the animals is required. Researchers prioritized the procurement of robust data to test hypotheses, analyze phenomena, and publish their results. Their concern for the animals rather reflected the views of the general public opposed to thought-out personal opinions on the matter; this is possibly due to their comparatively limited interaction with the animals. They considered the animals in abstract terms that were indicative of reification. This study concludes that animal reification is still present, albeit to varying degrees amongst the stakeholders.

Sheep enteric cestodes and their influence on clinical indicators used in targeted selective treatments against gastrointestinal nematodes.

Clinical indicators such as diarrhoea (DISCO) or anaemia (FAMACHA©) are used as a measure for targeted selective treatments against gastrointestinal nematodes (GIN). Enteric cestodes such as Moniezia may interfere directly with DISCO or indirectly with the FAMACHA© score. We investigated 821 Ouled Djellal rams naturally infected in a steppe environment (GIN alone, cestodes alone, GIN and cestodes) or not. The rams were treated with ivermectin 2 months before being slaughtered to reduce the impact of nematodes on the clinical scores; however, persistent or newly acquired GINs were not related to both scores. Of the non-infected rams (n = 296), 26% identified as needing treatment against GIN using the FAMACHA score, and 34.5% using DISCO would have been thus selected. This implies that the clinical indicators used for the targeted selective treatment of gastrointestinal nematodes are not fully reliable when a low infection is recorded and may well be influenced by confounding factors. As expected, only DISCO was affected by cestode infection, and we suggest that the presence of Moniezia should also be taken into consideration.

Identification of first-stage dorsal-spined lungworm larvae of Tunisian barbary red deer: First report of Varestrongylus sagittatus and Elaphostrongylus cervi in Africa.

Barbary red deer (Cervus elaphus barbarus) is a protected rare subspecies of red deer. The study of its Protostrongylidae fauna based only on sporadic necropsy of naturally dead animals is difficult. Therefore diagnosis of lungworms rely mainly on the identification of the first stage larvae (L1). The L1 of the different species are not readily diagnosed on morphological basis since much variation is recorded within and among dorsal-spined larvae belonging to various species. The aim of this study was to identify the dorsal-spined lungworm larvae of the Barbary red deer. A discriminant function was established, using the measurements of L1 lungworms recorded from red deer in the literature, then applied to identify 220 dorsal-spined larvae extracted from 25 Tunisian Barbary red deer fresh fecal samples. Also the ITS2 region of rDNA of four pools of larvae (n = 25-60) were amplified, sequenced and analyzed. Using discriminant analysis of morphological traits, Elaphostrongylus cervi and Varestongylus sagittatus were identified. Molecular identification confirmed the presence of E. cervi which is the most prevalent species. This study represents the first identification of V. sagittatus and E. cervi in North Africa.

Meta-analysis of the parasitic phase traits of Haemonchus contortus infection in sheep.

BACKGROUND: The parasitic nematode Haemonchus contortus shows highly variable life history traits. This highlights the need to have an average estimate and a quantification of the variation around it to calibrate epidemiological models. METHODS: This paper aimed to quantify the main life history traits of H. contortus and to identify explanatory factors affecting these traits using a powerful method based on a systematic review and meta-analysis of current literature. The life history traits considered are: (i) the establishment rate of ingested larvae; (ii) the adult mortality rate; (iii) the fertility (i.e. the number of eggs laid/female/day); and (iv) fecundity of female worms (i.e. the number of eggs per gram of faeces). RESULTS: A total of 37 papers that report single experimental infection with H. contortus in sheep and published from 1960 to 2015, were reviewed and collated in this meta-analysis. This encompassed 115 experiments on 982 animals. Each trait was analysed using a linear model weighted by its inverse variance. The average (± SE) larval establishment rate was 0.24 ± 0.02, which decreased as a function of the infection dose and host age. An average adult mortality rate of 0.021 ± 0.002) was estimated from the literature. This trait varied as a function of animal age, breed and protective response due to prior exposure to the parasite. Average female fertility was 1295.9 ± 280.4 eggs/female/day and decreased in resistant breeds and previously infected hosts. Average faecal egg count at necropsy was 908.5 ± 487.1 eggs per gram of faeces and varied as a function of infection duration and host resistance. The average sex ratio of H. contortus was 0.51 ± 0.006. CONCLUSION: This work is the first systematic review to summarise the available information on the parasitic phase of H. contortus in sheep. The results of the meta-analysis provide robust estimates of life history traits for parametrization of epidemiological models, their expected variation according to experimental factors, and provides correlations between these.

Ivermectin failure in the control of Oxyuris equi in a herd of ponies in France.

Drug resistance in equine gastro-intestinal parasitic nematodes has been reported throughout the world. While the focus is usually put on cyathostomins, observations of macrocylic lactone failure against Oxyuris equi have accumulated over the last decade. Here we report the failure of ivermectin in the control of O. equi in an experimental Welsh pony herd. In a first trial, 6 ponies previously drenched with moxidectin and showing patent O. equi infections were administered ivermectin and subsequently monitored for O. equi egg excretion over one month. This trial demonstrated a failure of ivermectin to control O. equi egg excretion as half of ponies demonstrated recurrent egg excretion in the peri-anal region during 21days after treatment. One year later, six female Welsh ponies drenched with moxidectin demonstrated signs of itching and scratching in their peri-anal region with worms being found transiently in fecal materials three weeks later. Ponies were allocated to three treatment groups, i.e. ivermectin, pyrantel embonate and fenbendazole and monitored for egg excretion over five weeks. Fenbendazole and pyrantel embonate broke ivermectin suboptimal efficacy as soon as 8 and 14days respectively after treatment, while egg excretion remained constant throughout the 41-day long trial in the ivermectin-treated ponies. This is the first report of ivermectin failure against O. equi in France. In the absence of critical efficacy test, it remains unclear whether true resistance is at stake or if these observations confound a constitutive suboptimal efficacy of ivermectin against O. equi.

Arrested development of abomasal trichostrongylid nematodes in lambs in a steppe environment (North-Eastern Algeria).

Arrested development of abomasal trichostrongylid nematodes was studied in 30 permanent grazing lambs on a large farm in the North-East of Algeria. The steppe climate has cold winters and hot and dry summers. The lambs were monitored monthly for gastrointestinal nematodes using nematode faecal egg counts, from February 2008 to February 2009. Every 2 months, two of the original 30 permanent lambs were necropsied after being held in pens for three weeks so that recently ingested infective larvae could develop into adults. The highest percentage of fourth stage larvae (L4), reaching 48% of the total worm burden, was recorded in abomasal contents in June. Teladorsagia and other Ostertagiinae constituted the highest percentage of L4 larvae (71%), whereas the percentage of Trichostrongylus (17.4%) or Haemonchus (11.6%) remained low. The dynamics of infection observed here (highest faecal egg count in August) and the stage composition of worm burden (highest percentage of L4 in June) provide strong evidence that arrested development had occurred.

Genetic relationship between the Echinococcus granulosus sensu stricto cysts located in lung and liver of hosts.

G1 genotype of Echinococcus granulosus sensu stricto is the major cause of hydatidosis in Northern Africa, Tunisia included. The genetic relationship between lung and liver localization were studied in ovine, bovine and human hydatid cysts in Tunisia. Allozyme variation and single strand conformation polymorphism were used for genetic differentiation. The first cause of genetic differentiation was the host species and the second was the localization (lung or liver). The reticulated genetic relationship between the liver or the lung human isolates and isolates from bovine lung, is indicative of recombination (sexual reproduction) or lateral genetic transfer. The idea of two specialized populations (one for the lung one for the liver) that are more or less successful according to host susceptibility is thus proposed.

A survey on parasite management by equine veterinarians highlights the need for a regulation change.

INTRODUCTION: In-depth knowledge of the use of anthelminthics in the field, especially by veterinarians, is required to design more sustainable parasite control strategies. MATERIALS AND METHODS: An online survey was sent by e-mail to 940 equine veterinary practitioners to describe their equine practice, their awareness about parasites and the management strategies they apply. RESULTS: Gastrointestinal parasites were generally considered (68%) as an issue of moderate importance. Drug efficacy failure was a minor or moderate issue for 47% and 48% of responders, respectively. Parasite management mostly relied on the use of systematic calendar treatments across a wide variety of horse owners (ie, riding schools, studs or hobby horse owners). Almost half of the practitioners (42%) never performed Faecal Egg Count (FEC) before drenching. Horse owners or their employees in charge of equines were reported to be the only person managing drenching in 59% of the collected answers. This was associated with the report of many off-label uses of anthelmintics and the frequent buying of drugs using the internet. CONCLUSIONS: Given the critical situation regarding anthelmintic resistance, it seems necessary for veterinarians to reclaim parasite management and prevention as a specific topic. Implementation of stricter regulations for use of anthelmintics, like the one applied in Denmark, may make parasitic management in equids more sustainable.

Economic assessment of FEC-based targeted selective drenching in horses.

In the face of an increased prevalence of drug-resistant cyathostomin populations, a targeted selective treatment (TST) strategy based on Faecal Egg Counts (FECs) has been proposed as an alternative management strategy. However, associated costs may be a barrier to the uptake of this strategy. Our study aims to provide an economic assessment of FEC-based TST. FECs were determined in a Welsh pony herd thrice a year from 2010 to 2014. This database was used to explore the impact of FEC price, sampling strategy (individual or pooled) and labour-associated costs. Drug price was set at the cheapest level, hence providing a conservative framework to determine the maximum viable FEC price in the context of a cost-driven horse industry. The maximum viable FEC price for a cost-efficient individual based strategy was determined by an in silico bootstrap approach consisting of randomly sampling 1000 virtual pony herds of various sizes (1 to 100 ponies) from the available database and estimating the associated costs (FEC price ranging from € 1 to € 10, anthelmintic costs and labour-associated costs). The costs and benefits of the pooling strategy that consists of basing the decision to treat on group FEC values were also investigated. This is thought to reduce FEC-based costs but may result in highly infected individuals being left undrenched, i.e. in false-negatives, as a result of FEC overdispersion. For various pool-sizes (1-20 ponies) and various cut-off thresholds (50-200 eggs/g), we sampled 1000 pony herds in silico to estimate the associated costs and determine the number of positive ponies within a negative pool. Following these simulations, pool-based FECs of various sizes were performed on 40 ponies to compare predictions with real data. Within 4 years, anthelmintic costs were cut by 80%, albeit with free FECs. In silico estimations suggested that an individual FEC-based TST would not be cost-efficient in this context for an FEC price above € 5. With a pooled FEC strategy, the proportion of false-negatives never exceeded 15% of the pool size. The combination of a 14-pony pool and a cut-off value of 150 eggs/g minimized total costs while keeping the number of false negatives to a reasonable level. Real data obtained from infected ponies however, suggested that pool size should not exceed 10 individuals, since the inhomogeneous mixing of faeces of larger pools probably reduced the correlation between average pooled FECs and the mean of individual FECs. Our study provides an economic framework that could be valuable for emphasizing the use of FEC-based approaches in the field.

Infection of dogs with Echinococcus granulosus: causes and consequences in an hyperendemic area.

BACKGROUND: Tunisia is a hyper endemic country for human echinococcosis. The infection is transmitted via the eggs of Echinococcus granulosus which are passed in the faeces of the definitive canid host. METHODS: This study evaluated the contamination rate of the dog faeces in different climatic conditions at eight different geographic regions throughout Tunisia. Dog faecal samples were collected from the soil and the Echinococcus eggs were identified using microscopic and molecular (Eg1121/1122 PCR, Egss1 PCR and Nad1 PCR-RFLP) tools. RESULTS: The contamination index of dog faeces by E. granulosus eggs ranged from 8.3% to 41.3% depending on the region. Comparisons of the dog faecal contamination rate against human incidence found them to be independent. Neither human prevalence nor dog contamination index appeared to be related to climatic conditions or geographic characteristics. The genetic variability of E. granulosus samples was different within each region but was not related to geographic distance which is indicative of local divergent evolutions rather than isolation by distance. CONCLUSIONS: A high environmental dog contamination index does not necessarily correspond to high prevalence in humans as transmission is strongly linked to human behavior and hygiene.

Estimation of genetic parameters for resistance to gastro-intestinal nematodes in pure blood Arabian horses.

Equine internal parasites, mostly cyathostomins, affect both horse welfare and performance. The appearance of anthelmintic-resistant parasites creates a pressing need for optimising drenching schemes. This optimization may be achieved by identifying genetic markers associated with host susceptibility to infection and then to drench carriers of these markers. The aim of our study was to characterise the genetics of horse resistance to strongyle infection by estimating heritability of this trait in an Arabian pure blood population. A population of 789 Arabian pure blood horses from the Michalów stud farm, Poland were measured for strongyle egg excretion twice a year, over 8 years. Low repeatability values were found for faecal egg counts. Our analyses showed that less than 10% of the observed variation for strongyle faecal egg counts in this population had a genetic origin. However, additional analyses highlighted an age-dependent increase in heritability which was 0.04 (±0.02) in young horses (up to 3 years of age) but 0.21 (±0.04) in older ones. These results suggest that a significant part of the inter-individual variation has a genetic origin. This paves the way to a genomic dissection of horse-nematode interactions which might provide predictive markers of susceptibility, allowing individualised drenching schemes.

Investigation of acetylcholine receptor diversity in a nematode parasite leads to characterization of tribendimidine- and derquantel-sensitive nAChRs.

Nicotinic acetylcholine receptors (nAChRs) of parasitic nematodes are required for body movement and are targets of important "classical" anthelmintics like levamisole and pyrantel, as well as "novel" anthelmintics like tribendimidine and derquantel. Four biophysical subtypes of nAChR have been observed electrophysiologically in body muscle of the nematode parasite Oesophagostomum dentatum, but their molecular basis was not understood. Additionally, loss of one of these subtypes (G 35 pS) was found to be associated with levamisole resistance. In the present study, we identified and expressed in Xenopus oocytes, four O. dentatum nAChR subunit genes, Ode-unc-38, Ode-unc-63, Ode-unc-29 and Ode-acr-8, to explore the origin of the receptor diversity. When different combinations of subunits were injected in Xenopus oocytes, we reconstituted and characterized four pharmacologically different types of nAChRs with different sensitivities to the cholinergic anthelmintics. Moreover, we demonstrate that the receptor diversity may be affected by the stoichiometric arrangement of the subunits. We show, for the first time, different combinations of subunits from a parasitic nematode that make up receptors sensitive to tribendimidine and derquantel. In addition, we report that the recombinant levamisole-sensitive receptor made up of Ode-UNC-29, Ode-UNC-63, Ode-UNC-38 and Ode-ACR-8 subunits has the same single-channel conductance, 35 pS and 2.4 ms mean open-time properties, as the levamisole-AChR (G35) subtype previously identified in vivo. These data highlight the flexible arrangements of the receptor subunits and their effects on sensitivity and resistance to the cholinergic anthelmintics; pyrantel, tribendimidine and/or derquantel may still be effective on levamisole-resistant worms.

Selective effect of the anthelmintic bephenium on Haemonchus contortus levamisole-sensitive acetylcholine receptors.

Acetylcholine receptors (AChRs) are pentameric ligand-gated ion channels involved in the neurotransmission of both vertebrates and invertebrates. A number of anthelmintic compounds like levamisole and pyrantel target the AChRs of nematodes producing spastic paralysis of the worms. The muscle AChRs of nematode parasites fall into three pharmacological classes that are preferentially activated by the cholinergic agonists levamisole (L-type), nicotine (N-type) and bephenium (B-type), respectively. Despite a number of studies of the B-type AChR in parasitic species, this receptor remains to be characterized at the molecular level. Recently, we have reconstituted and functionally characterized two distinct L-AChR subtypes of the gastro-intestinal parasitic nematode Haemonchus contortus in the Xenopus laevis oocyte expression system by providing the cRNAs encoding the receptor subunits and three ancillary proteins (Boulin et al. in Br J Pharmacol 164(5):1421-1432, 2011). In the present study, the effect of the bephenium drug on Hco-L-AChR1 and Hco-L-AChR2 subtypes was examined using the two-microelectrode voltage-clamp technique. We demonstrate that bephenium selectively activates the Hco-L-AChR1 subtype made of Hco-UNC-29.1, Hco-UNC-38, Hco-UNC-63, Hco-ACR-8 subunits that is more sensitive to levamisole than acetylcholine. Removing the Hco-ACR-8 subunit produced the Hco-L-AChR2 subtype that is more sensitive to pyrantel than acetylcholine and partially activated by levamisole, but which was bephenium-insensitive indicating that the bephenium-binding site involves Hco-ACR-8. Attempts were made to modify the subunit stoichiometry of the Hco-L-AChR1 subtype by injecting five fold more cRNA of individual subunits. Increased Hco-unc-29.1 cRNA produced no functional receptor. Increasing Hco-unc-63, Hco-unc-38 or Hco-acr-8 cRNAs did not affect the pharmacological characteristics of Hco-L-AChR1 but reduced the currents elicited by acetylcholine and the other agonists. Here, we provide the first description of the molecular composition and functional characteristics of any invertebrate bephenium-sensitive receptor.

Resistance of Trichostrongylus spp. (Nematoda) to benzimidazole in Algerian cattle herds grazed with sheep.

Drug resistance in cattle strongyles to anthelmintics is rarely described in Europe and in the Mediterranean areas. The faecal egg counts are usually low in cattle, and detection of resistance may be particularly difficult. We used the method of twice-repeated treatments and used various estimations of efficacies in order to detect cattle strongyle resistance to benzimidazoles. Resistance was found in two farms among eight that were studied. Trichostrongylus axei was the resistant species in one farm whereas Trichostrongylus sp. was found in another farm. In both farms, there were sheep flocks with a history of strongyle benzimidazole resistance and practice of alternate or mixed grazing with cattle.

The detection of snail host habitats in liver fluke infected farms by use of plant indicators.

Field investigations in 361 liver fluke infected cattle- or sheep-breeding farms on acid soil were carried out during thirty years in March and April to record indicator plants in relation to the category of site colonized by the intermediate host of liver fluke, the snail Galba truncatula. Seven types of snail zones and six species of indicator plants were recorded in the 7709 positive sites studied. The most frequent habitats were located at the peripheral extremities of open drainage furrows. Juncus acutiflorus, Juncus effusus, Glyceria fluitans, and Agrostis stolonifera were the indicator plants. Plant indicators were highly efficient (38-80% of variance explained) when used with the size of site area within a hydrographical zone. The identification of positive sites for G. truncatula is a key for controlling the intermediate host on pastures through biological control with predatory molluscs. The positive sites may be detected using a two-step method including first categorization of hydrographical zones and then, within a zone, use of one or several indicator plants (frequently J. acutiflorus).