First report of eprinomectin-resistant isolates of Haemonchus contortus in 5 dairy sheep farms from the Pyrénées Atlantiques département in France.

Infection of sheep by gastrointestinal nematodes (GIN) in pastoral systems such as those found in the South Western area of France, the Pyrénées Atlantiques, is one of the main reasons for economic loss and degradation of their welfare. In the present study, the efficacy of eprinomectin (EPN) was monitored on farms from this area following suspicion of lack of anthelmintic efficacy. Suspicions were raised by veterinarians, based on clinical signs ranging from milk and body condition loss, to anaemia, and mortality. Resistance was evaluated according to the World Association for the Advancement for Veterinary Parasitology (WAAVP) guidelines using fecal egg count reduction tests reinforced by individual analysis of drug concentration in the serum of all treated ewes by high-performance liquid chromatography (HPLC). EPN was administered by subcutaneous (SC) and topical (T) route according to manufacturer's requirements, as well as by the oral route (O) with the topical solution according to off-labelled practices in the field. For the first time in France, the presence of resistant isolates of Haemonchus contortus to EPN was observed in 5 dairy sheep farms. The HPLC dosages showed exposure of worms to concentrations compatible with anthelmintic activity for animals treated by the SC and O routes. By contrast, they showed under exposure to the drug of most individuals treated by the T route. EPN is the only null milk withdrawal anthelmintic molecule currently available. The presence of resistant isolates of the pathogenic H. contortus to EPN in this important dairy region requires an urgent change in grazing, and sometimes production, systems.

Stomoxys calcitrans, mechanical vector of virulent Besnoitia besnoiti from chronically infected cattle to susceptible rabbit.

Cattle besnoitiosis caused by Besnoitia besnoiti (Eucoccidiorida: Sarcocystidae) is a re-emerging disease in Europe. Its mechanical transmission by biting flies has not been investigated since the 1960s. The aim of this study was to re-examine the ability of Stomoxys calcitrans (Diptera: Muscidae) to transmit virulent B. besnoiti bradyzoites from chronically infected cows to susceptible rabbits. Three batches of 300 stable flies were allowed to take an interrupted bloodmeal on chronically infected cows, followed by an immediate bloodmeal on three rabbits (Group B). A control group of rabbits and a group exposed to the bites of non-infected S. calcitrans were included in the study. Blood quantitative polymerase chain reaction (qPCR) analyses, and clinical, serological and haematological surveys were performed in the three groups over 152 days until the rabbits were killed. Quantitative PCR analyses and histological examinations were performed in 24 tissue samples per rabbit. Only one rabbit in Group B exhibited clinical signs of the acute phase of besnoitiosis (hyperthermia, weight loss, regenerative anaemia and transient positive qPCR in blood) and was seroconverted. Parasite DNA was detected in four tissue samples from this rabbit, but no cysts were observed on histological examination. These findings indicate that S. calcitrans may act as a mechanical vector of B. besnoiti more efficiently than was previously considered.

A real-time PCR approach to identify anthelmintic-resistant nematodes in sheep farms.

Resistance to fenbendazole, ivermectin, and moxidectin was explored by a fecal egg count reduction test in four meat sheep flocks in southwestern France where anthelmintic resistance was suspected. The FECR test results of the present study confirmed the presence of benzimidazole resistance in three out of the four farms and the presence of ivermectin resistance in one flock. In addition, a suspicion of moxidectin resistance was shown in this latter farm. Both conventional morphological and molecular identifications were performed on larval cultures before and after the treatment in the studied farms. A high positive correlation was found between the number of larvae counted under binocular microscope and the number of larvae estimated by the qPCR analysis (R 2 = 0.88) and a high Cohen's Kappa value (0.91) in the detection of strongylid larvae in larval cultures. According to qPCR results, Trichostrongylus species demonstrated high levels of BZ resistance and Teladorsagia circumcincta was involved in the IVM resistance in one farm. The molecular procedures used in this study have the potential to be beneficial for anthelmintic resistance surveillance in sheep industry.

No detection of Besnoitia besnoiti DNA in the semen of chronically infected bulls.

Bovine besnoitiosis is a chronic and debilitating disease observed in many European countries that may cause important economic losses in cattle. The recent widespread of the parasite in Europe had led the European Food Safety Authority to declare bovine besnoitiosis as a re-emerging disease in Europe. Many aspects of the epidemiology of bovine besnoitiosis such as the main routes of transmission are still unclear and need to be further studied. Among the different hypotheses, a sexual transmission has not yet been investigated. Therefore, the aim of this study was to evaluate the presence of Besnoitia besnoiti DNA in the semen of naturally infected bulls by using a highly sensitive method (real-time qPCR). Both pre-sperm and sperm fractions of 40 bulls, including seronegative (n = 11), seropositive subclinically (n = 17), and seropositive clinically (n = 12) infected animals, were collected by electroejaculation and analyzed by real-time qPCR. No B. besnoiti DNA was detected in 27 pre-sperm and 28 sperm fractions of the 40 examined bulls, suggesting that the transmission of B. besnoiti infection by the semen of chronically infected bulls is very unlikely.

Development of a protocol testing the ability of Stomoxys calcitrans (Linnaeus, 1758) (Diptera: Muscidae) to transmit Besnoitia besnoiti (Henry, 1913) (Apicomplexa: Sarcocystidae).

Cattle besnoitiosis due to the cyst-forming coccidian parasite Besnoitia besnoiti has recently been reported in expansion in Europe since the end of the twentieth century. The B. besnoiti life cycle and many epidemiological traits are still poorly known. Hematophagous flies, including the worldwide-distributed Stomoxys calcitrans, could be mechanical vectors in the contamination of mouthparts after the puncture of cutaneous cysts or ingestion of infected blood. In this study, a protocol is presented to assess more deeply the role of S. calcitrans, reared in laboratory conditions, in parasite transmission. A preliminary trial showed that stable flies could transmit tachyzoites from bovine artificially parasite-enriched blood to B. besnoiti-free blood using glass feeders. Evidence of transmission was provided by the detection of parasite DNA with Ct values ranging between 32 and 37 in the blood recipient. In a second time, a B. besnoiti-infected heifer harboring many cysts in its dermis was used as a donor of B. besnoiti. An interruption of the blood meal taken by 300 stable flies from this heifer was performed. Immediately after the blood meal was interrupted, they were transferred to a glass feeder containing B. besnoiti-free blood from a non-infected heifer. Quantitative PCR and modified direct fluorescence antibody test (dFAT) were used to detect B. besnoiti DNA and entire parasites, respectively, in the blood recipient, the mouthparts, and the gut contents of S. calcitrans at two time intervals: 1 and 24 h after the interrupted blood meal. Parasite DNA was detected at both time intervals (1 and 24 h) in all samples (blood recipient, mouthparts, and gut contents of stable flies) while entire parasites by dFAT were only found in the abdominal compartment 1 h after the interrupted blood meal. Then, S. calcitrans were able to carry B. besnoiti from chronically infected cattle to an artificial recipient in the conditions of the protocol.

Cell grazing and Haemonchus contortus control in sheep: lessons from a two-year study in temperate Western Europe.

Managing infections of sheep with anthelmintic resistant gastrointestinal nematodes (GIN) is a major challenge for sheep producers in Western Europe. New methods of grazing management have been poorly explored as a component of an integrated and sustainable control of these parasites. Therefore, the purpose of this study was to evaluate the effect of two different types of grazing systems of sheep (intensive cell grazing versus conventional rotational grazing) on GIN infections over two years in a farm located in a temperate environment of Western France. When considering the whole study, the type of grazing system did not influence significantly the intensity of egg excretions of adult ewes even if the proportion of ewes excreting high numbers of GIN eggs was higher in cell grazing system than in rotational grazing system. The most striking result of this survey was the effect of grazing system on the GIN species composition harbored by ewes and by their lambs: with time, the proportions of H. contortus infections were lower in cell grazing system than in rotational grazing system. In conclusion, the cell grazing system, as implemented in this study, could limit the importance of this highly pathogenic nematode species in sheep.

Specific IgG antibody responses in Oestrus ovis L. (Diptera: Oestridae) infected sheep: associations with intensity of infection and larval development.

Larvae of Oestrus ovis (Diptera: Oestridae) are ubiquitous parasites of nasal and sinusal cavities of sheep and goats. According to the chronobiology of O. ovis infections in Sardinia and the seasonal pattern of the IgG response, the optimal period to investigate the relationships between O. ovis larval populations and intensity of local and systemic IgG antibody responses was mid-July in the summer season. Sarda x Lacaune ewes (n=186), divided into three ram-families were used in the study. Systemic and local IgG responses were measured by ELISA tests using second stage larval crude extracts (L2CE) and L2 (L2SGC) and L3 (L3SGC) salivary gland contents as coating antigens. The number of larval instars, larval length of L1, L2 and L3 larvae, and larval weight of L2 and L3 larvae were individually recorded after ewe necropsy. Negative correlations among larval establishment and/or larval development on the one hand and intensity of local or systemic IgG responses on the other hand were found in two out of three studied ram-families.

Multiple-resistance to ivermectin and benzimidazole of a Haemonchus contortus population in a sheep flock from mainland France, first report.

Anthelmintic resistance was explored by fecal egg count reduction test in a sheep flock from the French Pyrenees at the request of the veterinary practitioner after a poor response to anthelmintics was noted. The FECRT confirmed the suspicion with a mean percentage of reduction in egg excretions of 45% (CI 95%: - 40 to 78.5) and 0% (CI95%: - 162 to 49) within the ivermectin and the benzimidazole groups respectively. Haemonchus contortus was shown to be the IVM and BZ resistant species after morphological and molecular characterizations whereas Teladorsagia circumcincta was probably resistant to BZ only. The H. contortus population was still susceptible to moxidectin, closantel and levamisole. As this sheep flock is a transhumant flock, the spread of this multiple-resistant Haemonchus contortus population to the other sheep flocks sharing the same pastures in Pyrenean Mountains is highly likely. From the knowledge of the authors, this is the first report of multi-resistance to ivermectin and benzimidazole of a Haemonchus contortus population in mainland France.

Resistance to gastrointestinal nematodes in dairy sheep: Genetic variability and relevance of artificial infection of nucleus rams to select for resistant ewes on farms.

Breeding sheep for enhanced resistance to gastrointestinal parasites is a promising strategy to limit the use of anthelmintics due to the now widespread resistance of parasites to these molecules. This paper reports the genetic parameters estimated for parasite resistance and resilience traits in the Blond-faced Manech dairy sheep breed and the putative impacts of the selection for resistance to gastrointestinal nematodes (GIN) on farms. Two datasets were used. First, the rams of the selection scheme were artificially infected twice with L3 Haemonchus contortus larvae. Faecal egg counts (FEC) and packed cell volume (PCV) loss were measured 30 days after each infection. Secondly, the FEC, PCV and body condition score (BCS) (1-6 measures per ewe) of naturally infected ewes on farms were measured in the spring, summer and autumn over a two-year period. Genetic parameters were estimated for each dataset independently but also globally based on the pedigree connections between the two datasets. For the experimentally infected sires, the FEC following the second infection was moderately heritable (heritability: 0.35) and strongly correlated with FEC after the first infection (genetic correlation: 0.92). For the naturally infected ewes, FEC was also heritable (0.18). Using the two datasets together, a genetic correlation of 0.56-0.71 was estimated between the FEC values of the experimentally infected rams and naturally infected ewes. Consequently, the genetic variability of parasite resistance is similar whatever the physiological status (males or milking/pregnant ewes) and the infection conditions (experimental infection with one parasite or natural infection with several parasites). In practice, when the sire population is divided into two groups based on their genetic value, the FEC of the ewes born to the 50% most resistant sires is half that of the ewes born to the 50% most susceptible sires. Our study shows the feasibility and efficiency of genetic selection for parasitism resistance based on the sires' FEC records to improve parasite resistance in naturally grazing ewes. For breed improvement, and to increase the selection pressure on parasite resistance, it seems more appropriate to measure FEC values on rams after experimental infection rather than on ewes in natural infection conditions because this limits the number and standardizes the conditions of FEC measurements.

Experimental concurrent infection of sheep with Oestrus ovis and Trichostrongylus colubriformis: effects of antiparasitic treatments on interactions between parasite populations and blood eosinophilic responses.

The purpose of this experiment was to determine if an earlier infection with Oestrus ovis would down regulate an infection with Trichostrongylus colubriformis when the larvae of O. ovis were expelled from the nasal cavities of sheep by a specific treatment. Three groups of five lambs were used: group 1 was artificially infected with O. ovis larvae and later with T. colubriformis, group 2 received O. ovis larvae and later was treated with ivermectin 14 days before being infected with T. colubriformis. Group 3 was infected with T. colubriformis only. The criteria examined were: the effects on nematode egg excretion, worm fecundity, nematode burdens and the kinetics of blood eosinophils. Significant decreases of nematode egg excretion, worm fecundity, nematode burdens were observed in group 1 compared to group 3. However, no changes were observed in either group 2 or 3. In group 2 it was noted that antiparasitic treatment induced a rapid decrease in blood eosinophils to a range close to the non-infected control group and this was associated with the removal of the down regulation effects of nematode burdens. This experiment showed that there is no cross immunity between O. ovis and T. colubriformis and that eosinophils may act against any parasite without specific priming.

Does bovine besnoitiosis affect the sexual function of chronically infected bulls?

Bovine besnoitiosis is a reemerging disease in Europe. The clinically Besnoitia besnoiti infection in bulls is characterized by fever, nasal discharge, and orchitis in the acute phase and by scleroderma in the chronic phase. However, in many bulls, B besnoiti infection remains at a subclinical stage. Bull infertility is an economically relevant consequence of besnoitiosis infection. It is not clear, however, if semen quality returns to normal levels when infected animals have clinically recovered. The aim of this study was to examine the relationship between chronic besnoitiosis and bull sexual function in a region of eastern France, where the disease is reemerging, by comparing semen quality and genital lesions in 11 uninfected, 17 subclinically infected, and 12 clinically infected bulls. The presence of anti-B besnoiti antibodies was detected by Western blot test. Semen was collected by electroejaculation. Bulls clinically infected with B besnoiti showed significantly more genital tract alterations than uninfected or subclinically infected bulls. No relationship was evidenced between besnoitiosis infectious status and semen quality, whereas a significant relationship was noted between genital lesions and semen score. This means that in the absence of moderate to severe genital lesions, chronic bovine besnoitiosis is unlikely to alter semen quality. However, as the presence of infected animals could lead to spread of the disease, culling or separation of clinically infected bulls from the remaining healthy animals is strongly recommended.

Haemonchus contortus egg excretion and female length reduction in sheep previously infected with Oestrus ovis (Diptera: Oestridae) larvae.

Mixed parasitic infection of animals is a common phenomenon in nature. The existence of one species often positively or negatively influences the survival of the other. Our experimental study was started with the objectives to demonstrate the interaction of Haemonchus contortus and Oestrus ovis in relation to cellular and humoral immune responses in sheep. Twenty-two sheep of Tarasconnais breed (France) were divided into four groups (O, OH, H and C) of five or six animals. Group O and OH received 5 weekly consecutive inoculations with O. ovis L1 larvae (total = 82 L1) in the first phase of the experiment between days 0 and 28. On the second phase, groups OH and H received 5000 L3 of H. contortus on day 48 while group C served as our control throughout the experimental period. Parasitological, haematological, serological and histopathological examinations were made according to standard procedures and all animals were slaughtered at day 95. There was no significant variation in the number and degree of development of O. ovis larvae between the two infected groups. Furthermore, in tissues examined in the upper respiratory tract (nasal septum, turbinate, ethmoide and sinus), group O and OH has responded similarly on the basis of cellular inflammatory responses (blood and tissue eosinophils, mast cells and globule leucocytes (GL)) and serum antibody responses against the nasal bots. This may indicate that the presence of H. contortus in the abomasa of group OH had no marked influence over the development of O. ovis larvae in the upper respiratory tract. On the other hand, we have observed a significantly lower H. contortus female worm length, fecal egg count (FEC) and in utero egg count in animals harbouring the nasal bot (OH) than in the mono-infected group (H). This was significantly associated with higher blood eosinophilia, higher packed cell volume (PCV) and increased number of tissue eosinophils and globule leucocytes. We conclude that, the establishment of O. ovis larvae in the upper respiratory tract has initiated higher inflammatory cellular activity in group OH there by influencing the development and fecundity of H. contortus in the abomasum.

Effects of condensed tannins on goats experimentally infected with Haemonchus contortus.

Although the use of tanniferous plants or condensed tannins as an alternative to anthelmintics to control gastrointestinal nematodes has been largely documented in sheep, studies remain scarce in goats. The objective of this study was therefore to assess the possible impact of condensed tannins in goats infected with adult Haemonchus contortus. Two groups of cull goats were experimentally infected with 10.000 L3 of H. contortus. After 4 weeks, quebracho extracts, representing 5% of the diet DM, were administered for 8 days to one of the two groups. Goats of the second group remained as controls. One week after the end of quebracho administration, the goats were euthanised. Individual egg excretion and pathophysiological parameters were measured weekly during the study. At the end of the study, worm counts were assessed and histological samples from the abomasa were taken to count the numbers of mucosal mast cells, globule leukocytes and eosinophils. The administration of tannins was associated with a significant decrease in egg excretion, which persisted until the end of experiment. This reduction was not associated with any difference in worm number but with a significant decrease in female fecundity. No significant changes in the mucosal density of the three inflammatory cell types were detected between the two groups. These results indicate that the major consequence of tannin consumption in goats is a reduction in worm fecundity and egg output, which does not seem related to significant changes in the local mucosal response.

Concurrent parasitic infections of sheep: depression of Trichostrongylus colubriformis populations by a subsequent infection with Oestrus ovis.

Concurrent infections of sheep with Oestrus ovis and trichostrongyles of the digestive tract are common in the field. Previous results have shown that a previous infection with O. ovis adversely affects worm populations of either Trichostrongylus colubriformis or Haemonchus contortus. However, no information was available to determine the influence of the succession of infections on the expression of interactions between these parasites located in remote anatomical sites. In order to investigate the role of these modulating factors, an experimental study was conducted on four groups of naïve sheep, examining the consequences of a delayed infection with O. ovis on a pre-existing population of T. colubriformis. group T was infected four times with 4000 T. colubriformis larvae on days 0, 14, 28 and 42 of experiment; group O received multiple infections with O. ovis first instar larvae on days 42, 49, 56, 70 and 77; sheep from group TO received both infections and animals from group C remained as uninfected controls. Faecal egg counts and eosinophilia were measured weekly throughout the study. At necropsy (day 91), the mucosal cellular responses in the nasal cavities (septum, turbinates, ethmoid and sinus) and in the digestive tract (stomach and small intestine) from all animals were analysed from histological sections. Infection of the digestive tract with nematodes did not modify the biology of Oestrus populations, as measured by the number and weight of larvae. In contrast, infections with O. ovis after T. colubriformis infection was related to significant reductions (P < 0.01) in nematode egg excretion and worm burdens. These changes were associated with significant modifications in populations of mast cells, globule leucocytes and eosinophils in the respiratory and digestive tracts. These results indicate that an antagonistic interaction exists between the populations of O. ovis in the nasal cavities and T. colubriformis in the small intestine but that the order of succession of infections with the two parasites is not a major modulating factor for expression of interactions. They also confirm that parasitic infection in one particular anatomical site induces "at distance" inflammatory reactions of the whole mucosal system.

The relationship between nasal myiasis and the prevalence of enzootic nasal tumours and the effects of treatment of Oestrus ovis and milk production in dairy ewes of Roquefort cheese area.

Infection by Oestrus ovis is common in Lacaune dairy ewes of Roquefort cheese area (Aveyron, France). It is believed by local breeders that there is a close relationship between nasal myiasis and the incidence of enzootic nasal tumour. In order to check these anecdotal reports, a serological survey was done on 658 breeding ewes before turn-out and 897 breeding and primiparous (hoggets) ewes at the end of the grazing season. By the time of sampling, it was clear whether the sheep were infected at the end of the winter or had been re-infected over summer. In April and September, 40.7 and 26.3%, respectively, were free of O. ovis infection, indicating that the autumn treatment was not completely effective and that O. ovis adult flies were circulating during the summer in many flocks. There were no differences in the incidence of adenocarcinoma between the groups indicating that there is no relationship between O. ovis infection and the presence of the cancer. Differences in milk production between the three groups were not statistically significant (Anova test P>0.05). In flocks where 1-5% of the ewes were infected or in non-infected flocks, ewes produced 3.6 and 8.56%, respectively, more milk than ewes from flocks where more than 5% of animals were infected. For primiparous ewes, the differences were of 8.5 and 12.24%.

Experimental concurrent infection of sheep with Oestrus ovis and Trichostrongylus colubriformis: negative interactions between parasite populations and related changes in the cellular responses of nasal and digestive mucosae.

Concurrent infections of sheep with Oestrus ovis and trichostrongyles of the digestive tract are common in the field. In order to examine the possible occurrence of interactions between these two parasites and the consequences on parasite biology and the associated pathophysiological changes, an experimental study was conducted on four groups of naïve sheep. Group O was infected repeatedly with O. ovis larvae from D0 to D42 of experiment; group T received a single infection of Trichostrongylus colubriformis on D49; sheep from group OT received both infections, and animals from group C remained uninfected as controls. Parasitological and pathophysiological parameters were measured regularly. At necropsy (D94), the mucosal cellular responses in the nasal cavities and digestive tract from all animals were analysed from histological sections. Infection of the digestive tract with nematodes did not modify the biology of Oestrus populations. In contrast, the presence of O. ovis was related to significant reductions in nematode egg excretion, worm fecundity and worm burdens. These changes were associated with significant modifications in tissular populations of mast cells, globule leucocytes and eosinophils in the respiratory and digestive tracts. These results indicate that parasitic infection in one particular anatomical site induces "at distance" inflammatory reactions of the whole mucosal system. The relationships between the cellular response and the changes in worm biology are discussed.

An inter-laboratory comparative study of serological tools employed in the diagnosis of Besnoitia besnoiti infection in bovines.

Bovine besnoitiosis is considered an emerging chronic and debilitating disease in Europe. Many infections remain subclinical, and the only sign of disease is the presence of parasitic cysts in the sclera and conjunctiva. Serological tests are useful for detecting asymptomatic cattle/sub-clinical infections for control purposes, as there are no effective drugs or vaccines. For this purpose, diagnostic tools need to be further standardized. Thus, the aim of this study was to compare the serological tests available in Europe in a multi-centred study. A coded panel of 241 well-characterized sera from infected and non-infected bovines was provided by all participants (SALUVET-Madrid, FLI-Wusterhausen, ENV-Toulouse, IPB-Berne). The tests evaluated were as follows: an in-house ELISA, three commercial ELISAs (INGEZIM BES 12.BES.K1 INGENASA, PrioCHECK Besnoitia Ab V2.0, ID Screen Besnoitia indirect IDVET), two IFATs and seven Western blot tests (tachyzoite and bradyzoite extracts under reducing and non-reducing conditions). Two different definitions of a gold standard were used: (i) the result of the majority of tests ('Majority of tests') and (ii) the majority of test results plus pre-test information based on clinical signs ('Majority of tests plus pre-test info'). Relative to the gold standard 'Majority of tests', almost 100% sensitivity (Se) and specificity (Sp) were obtained with SALUVET-Madrid and FLI-Wusterhausen tachyzoite- and bradyzoite-based Western blot tests under non-reducing conditions. On the ELISAs, PrioCHECK Besnoitia Ab V2.0 showed 100% Se and 98.8% Sp, whereas ID Screen Besnoitia indirect IDVET showed 97.2% Se and 100% Sp. The in-house ELISA and INGEZIM BES 12.BES.K1 INGENASA showed 97.3% and 97.2% Se; and 94.6% and 93.0% Sp, respectively. IFAT FLI-Wusterhausen performed better than IFAT SALUVET-Madrid, with 100% Se and 95.4% Sp. Relative to the gold standard 'Majority of test plus pre-test info', Sp significantly decreased; this result was expected because of the existence of seronegative animals with clinical signs. All ELISAs performed very well and could be used in epidemiological studies; however, Western blot tests performed better and could be employed as a posteriori tests for control purposes in the case of uncertain results from valuable samples.

A genome scan for QTL affecting resistance to Haemonchus contortus in sheep.

Gastrointestinal nematodes are one of the main health issues in sheep breeding. To identify loci affecting the resistance to Haemonchus contortus, a genome scan was carried out using 1,275 Romane × Martinik Black Belly backcross lambs. The entire population was challenged with Haemonchus contortus in 2 consecutive experimental infections, and fecal egg counts (FEC) and packed cell volumes were measured. A subgroup of 332 lambs with extreme FEC was necropsied to determine the total worm burden, length of female worms, sex ratio in the worm population, abomasal pH, and serum and mucosal G immunoglobulins (IgG) responses. Pepsinogen concentration was measured in another subset of 229 lambs. For QTL detection, 160 microsatellite markers were used as well as the Illumina OvineSNP50 BeadChip that provided 42,469 SNP markers after quality control. Linkage, association, and joint linkage and association analyses were performed with the QTLMAP software. Linkage disequilibrium (LD) was estimated within each pure breed, and association analyses were carried out either considering or not the breed origin of the haplotypes. Four QTL regions on sheep chromosomes (OAR)5, 12, 13, and 21 were identified as key players among many other QTL with small to moderate effects. A QTL on OAR21 affecting pepsinogen concentration exactly matched the pepsinogen (PGA5) locus. A 10-Mbp region affecting FEC after the 1st and 2nd infections was found on OAR12. The SNP markers outperformed microsatellites in the linkage analysis. Taking advantage of the LD helped to refine the locations of the QTL mapped on OAR5 and 13.

A longitudinal study of Besnoitia besnoiti infections and seasonal abundance of Stomoxys calcitrans in a dairy cattle farm of southwest France.

Bovine besnoitiosis, caused by the cyst-forming apicomplexan Besnoitia besnoiti, is commonly reported in some restricted regions of South-Western Europe, and in larger regions of Africa and Asia. This infection is thought to be transmitted by blood feeding insects and is responsible for major economic losses in cattle production. A recent emergence in Europe, notified in the Centre of France, Spain and Germany, has attracted more attention to this disease. Clinical signs could appear in some animals; however, many infected cattle remain asymptomatic or show scleral-conjunctival cysts (SCC) only. Recent development of serological methods allows carrying out seroepidemiological field studies. In this respect, a long-term investigation was performed in a dairy cattle farm localized in an enzootic area of besnoitiosis of South-western France between March 2008 and May 2009. The objective was to estimate the seasonal pattern of B. besnoiti infections based on the presence of SCC and serology (ELISA and Western blot). In parallel, an entomological survey was conducted to describe population dynamics of Stomoxys calcitrans and Tabanidae species. The seroprevalence determined by Western blot in a cohort of 57 animals continuously present during the whole survey increased from 30% in March 2008 to 89.5% in May 2009 and was always higher than the prevalence based on clinically assessed SCC. New positive B. besnoitia seroconversions occurred throughout the year with the highest number in spring. In addition, many seroconversions were reported in the two months before turn-out and could be associated with a high indoors activity of S. calcitrans during this period.

Eosinophils in Haemonchus contortus-infected resistant and susceptible breeds of sheep: abomasal tissue recruitment and in vitro functional state.

A comparative experimental study was performed between Barbados Black Belly (resistant) and INRA-401 (susceptible) breeds of sheep in which primary infection with Haemonchus contortus was terminated on day 16. Measurements of parasite burden, abomasal tissue eosinophilia, IL-4, IL-5 and IL-13 mRNA transcripts in abomasal mucosa, and in vitro larval killing abilities of blood eosinophils were performed. The results show that: (1) worm burden was significantly lower and blood eosinophilia higher in the Black Belly than in the INRA breed. (2) Abomasal cytokine expression was noticed but no difference existed between the two breeds. (3) Three out of four Black Belly sheep had higher tissue eosinophil numbers compared to the INRA sheep (more eosinophils observed in the pyloric than in the fundic region in both breeds). (4) No significant difference was observed in the in vitro larval immobilizing potential of eosinophils between the two breeds. Collectively, abomasal eosinophil number and larval killing abilities of blood eosinophils do not seem to explain the difference in worm burden between the two breeds.