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.

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.

Divergent selection on milk somatic cell count in goats improves udder health and milk quality with no effect on nematode resistance.

Milk somatic cell count (SCC) is commonly higher in goats than in cattle and sheep. Furthermore, the ability of milk SCC to predict mastitis is considered lower in goats than in cattle and sheep, and the relevance of somatic cell score (SCS)-based selection in this species has been questioned. To address this issue, we created 2 divergent lines of Alpine goats using artificially inseminated bucks with extreme estimated breeding values for SCS. A total of 287 goats, 158 in high- and 129 in low-SCS lines, were scrutinized for mastitis infections. We subjected 2,688 milk samples to conventional bacteriological analyses on agarose and bacterial counts were estimated for positive samples. The SCS, milk yield, fat content, and protein content were recorded every 3 wk. Clinical mastitis was systematically noted. A subset of 40 goats (20 from each line) was subsequently challenged with Haemonchus contortus and monitored for anemia (blood packed cell volume) and fecal egg counts to see if SCS-based selection had an indirect effect on resistance to gastrointestinal nematodes. Milk production traits, including milk quantity, fat content, and protein content, were similar in both goat lines. In contrast, the raw milk SCC almost doubled between the lines, with 1,542,000 versus 855,000 cells/mL in the high- and low-SCS lines, respectively. The difference in breeding value for SCS between lines was 1.65 genetic standard deviation equivalents. The Staphylococcus spp. most frequently isolated from milk were S. xylosus, S. caprae, S. epidermidis, and S. aureus. The frequency of positive bacteriology samples was significantly higher in the high-SCS line (49%) than in the low-SCS line (33%). The highest odds ratio was 3.49 (95% confidence interval: 11.95-6.25) for S. aureus. The distribution of bacterial species in positive samples between lines was comparable. The average quantity of bacteria in positive samples was also significantly higher in high-SCS goats (69 ± 80 growing colonies) than in low-SCS goats (38 ± 62 growing colonies). Clinical cases were rare and equally distributed between high- (n = 4; 2.5%) and low-SCS (n = 3; 2.3%) lines. Furthermore, the larger the amounts of bacteria in milk the higher the SCS level. Conversely, goats with repeatedly culture-negative udders exhibited the lowest SCC levels, with an average of below 300,000 cells/mL. We therefore confirmed that SCS is a relevant predictor of intramammary infection and hygienic quality of milk in goats and can be used for prophylactic purposes. After challenge with H. contortus, goats were anemic with high fecal egg counts but we found no difference between the genetic lines. This result provides initial evidence that resistance to mastitis or to gastrointestinal nematodes infections is under independent genetic regulation. Altogether, this monitoring of the goat lines indicated that SCS-based selection helps to improve udder health by decreasing milk cell counts and reducing the incidence of infection and related bacterial shedding in milk. Selection for low SCC should not affect a goat's ability to cope with gastrointestinal nematodes.

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.

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.

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.

Risk factor analysis of equine strongyle resistance to anthelmintics.

Intestinal strongyles are the most problematic endoparasites of equids as a result of their wide distribution and the spread of resistant isolates throughout the world. While abundant literature can be found on the extent of anthelmintic resistance across continents, empirical knowledge about associated risk factors is missing. This study brought together results from anthelmintic efficacy testing and risk factor analysis to provide evidence-based guidelines in the field. It involved 688 horses from 39 French horse farms and riding schools to both estimate Faecal Egg Count Reduction (FECR) after anthelmintic treatment and to interview farm and riding school managers about their practices. Risk factors associated with reduced anthelmintic efficacy in equine strongyles were estimated across drugs using a marginal modelling approach. Results demonstrated ivermectin efficacy (96.3% ± 14.5% FECR), the inefficacy of fenbendazole (42.8% ± 33.4% FECR) and an intermediate profile for pyrantel (90.3% ± 19.6% FECR). Risk factor analysis provided support to advocate for FEC-based treatment regimens combined with individual anthelmintic dosage and the enforcement of tighter biosecurity around horse introduction. The combination of these measures resulted in a decreased risk of drug resistance (relative risk of 0.57, p = 0.02). Premises falling under this typology also relied more on their veterinarians suggesting practitionners play an important role in the sustainability of anthelmintic usage. Similarly, drug resistance risk was halved in premises with frequent pasture rotation and with stocking rate below five horses/ha (relative risk of 0.53, p < 0.01). This is the first empirical risk factor analysis for anthelmintic resistance in equids. Our findings should guide the implementation of more sustained strongyle management in the field.

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.

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.

Efficacy of a new combination of fipronil and permethrin (Effitix®) against Phlebotomus perniciosus in dogs.

Two controlled clinical trials were carried out to assess the anti-feeding and adulticidal effects of a spot-on combining fipronil and permethrin (Effitix(®), Virbac, Carros, France) against Phlebotomus perniciosus in dogs. The first study (Exp. A) was a dose-determination study in which 3 doses of permethrin (30 mg/kg, 60 mg/kg and 120 mg/kg) were compared. The second study (Exp. B) was an efficacy study using commercial dose of permethrin contained in Effitix(®) (the minimum dose of permethrin applied to dogs was 60 mg/kg). Twenty four and twelve Beagle dogs with equal sensitivity to sandflies were included in Exp. A and in Exp. B, respectively. Dogs were challenged with female sandflies (50 per dogs in Exp. A and 80 in Exp. B) for 60±5 min on Days - 7, 1, 7, 14, 21 and 28 (Day 0 being treatment day). Counts and engorgement determination of dead and alive sandflies were performed after each exposure to treated and untreated dogs. Dead sandflies were also counted 24 h after exposure. In Exp. A, the repellency induced by an administration of 30 mg/kg of permethrin to dogs was above 91% for the first two weeks and then dropped to 82.2, 83.1 and 81.1% on Days 14, 21 and 28, respectively. For dogs receiving 60 mg/kg of permethrin, the repellency was a bit higher with 95.8, 97.6, 92.1, 91.4, and 86.8%, for Days 1, 7, 14, 21 and 28, respectively. The repellency induced by 120 mg/kg of permethrin was significantly higher than that induced by 60 mg/kg of permethrin on Day 14 only. In Exp. B the anti-feeding effect of the spot-on formulation was 94.1, 97.8, 96.3, 90.8 and 87% on Days 1, 7, 14, 21 and 28, respectively. The mortality effect was 98.9, 99.1, 99.8, 97.0 and 89.7% on Days 1, 7, 14, 21 and 28, respectively. At each challenge point, the mortality and anti-feeding effects on sandflies were significantly different between control and treatment group (p<0.05). The results indicate that a monthly administration of this new combination of permethrin and fipronil could be used as an effective sandfly control strategy in dogs and therefore recommended for use in an integrated leishmaniosis prevention program.

Storage of gastrointestinal nematode infective larvae for species preservation and experimental infections.

Techniques to preserve the infective third-stage larvae (L3) of gastrointestinal nematodes are of considerable interest to preserve rare species and to maintain a stable source for routine experimental infections. This study compares the relative pros and cons of the two most common techniques, cryopreservation and refrigeration by comparing how they influence consequent infection outcome parameters in terms of life-history traits and fitness as a function of time using the gastrointestinal nematode of sheep Haemonchus contortus as a study species. Establishment capacity was found to be significantly reduced in cryopreserved stocks of L3 compared to refrigerated stocks, but this was followed by significant increases in their fecundity. Refrigeration did not affect L3 stocks consequent fitness by 16 months (the maximum examined) although they did incur a significant reduction in establishment, followed once again by an augmentation in fecundity. The study highlights potential areas for bias in comparing studies using L3 larvae maintained for different periods of time under different techniques.

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.

Efficacy of dinotefuran, permethrin and pyriproxyfen combination spot-on on dogs against Phlebotomus perniciosus and Ctenocephalides canis.

This study was conducted to evaluate the efficacy of a new topical ectoparasiticidal spot-on containing 4.95% dinotefuran (w/w), 36.08% permethrin (w/w) and 0.44% pyriproxyfen (w/w) (Vectra 3D, Ceva, Libourne, France) against Portuguese strain of Phlebotomus perniciosus and a French strain of Ctenocephalides canis in dogs. Twelve beagle dogs were exposed for 1 h to 100 P. perniciosus on day 6 for allocation in two groups. One group was treated on day 0, and the other group was the control group. The dogs were exposed for 1 h to 100 P. perniciosus on days 1, 7, 14, 21 and 28. After each sandfly challenge, the same dogs were infested with 100 C. canis. Counts of living fleas were determined 48 h after infestation on days 4, 3, 9, 16, 23 and 30. For sandflies, the anti-feeding effect was 96.9, 99.7, 98.7, 83.5 and 87.0 % on days 1, 7, 14, 21 and 28, respectively. The mortality effect was 97.8, 99.8, 73.7, 27.5 and 39.6% on days 1, 7, 14, 21 and 28, respectively. At each challenge point, the mortality and anti-feeding effects on sandflies were significantly different between the control and treatment groups (p < 0.05). The adulticidal effect on C. canis remained above 99% throughout the study period. The results indicate that a combination with dinotefuran, permethrin and pyriproxyfen may be used as an effective part of an overall flea and sandfly control strategy in dogs for monthly use.

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.

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.

Feeding and breeding aspects of Stomoxys calcitrans (Diptera: Muscidae) under laboratory conditions.

Bionomic aspects of Stomoxys calcitrans (Linnaeus, 1758) (Diptera: Muscidae) were studied under laboratory conditions. For this reason, laboratory-rearing techniques were optimized at the National Veterinary School of Toulouse. The colony was maintained at 25±2°C, 50±10% RH under a 12-hour light cycle and observed daily. The size of each adult cage is 30x30x30 cm and designed to house about 500-1,000 flies. The average cycle from egg to adult was 19.2±1.7 days. The mean longevity of imagos was 9.3±5.8 days and not significantly different between sexes. Stable flies were split into two groups; the first was fed with blood, honey and water, and the second was fed only with honey and water. The mean weight of a blood meal was 11.1±3.8 mg with no significant differences between males and females. The mean longevity of non-blood fed flies was found to be significantly higher (10.4±3.9 days) than those fed with blood. The maximum lifespan was shorter for non-blood fed males (17 days) and females (18 days) than for those fed with blood (females: 24 days, males: 23 days). Under these laboratory conditions, S. calcitrans rearing was successfully established. In the end, the number of expected generations of S. calcitrans and the net reproduction rate were estimated to be 11.8 generations/year and 16.2 living females per female respectively.

Efficacy of permethrin, dinotefuran and pyriproxyfen on adult fleas, flea eggs collection, and flea egg development following transplantation of mature female fleas (Ctenocephalides felis felis) from cats to dogs.

A novel spot-on formulation combining permethrin, pyriproxifen and dinotefuran (Vectra 3D™ spot-on solution for dogs) was evaluated in adult Beagle dogs in a study to determine adulticidal efficacy, egg laying inhibition and viability of Ctenocephalides felis felis eggs (development and emergence of fleas from the collected eggs). Prior to treatment sixteen dogs were checked for their ability to keep fleas 24 hours after infestation and were allocated to treatment groups: 8 dogs served as untreated controls, and 8 dogs were treated once with the tested formulation. The spot on was administered respecting the laboratory recommendations at a dosage of 65-126 mg/kg of permethrin; 8.9-17.4 mg/kg of dinotefuran and 0.8-1.5mg/kg of pyriproxyfen. Each dog was infested with 100 adult cat fleas ready to lay eggs after 72 hours spent feeding on cats. Dogs were infested 24 hours after treatment and then weekly during 63 days. Eggs were collected and counted 24 hours after each infestation and dogs were combed 48 hours after each infestation. Fleas were counted and removed. Collected eggs were placed in incubator to study their development in larvae and into newly emerged adults. A single treatment provided 99.7% adulticidal efficacy on fleas within 48 hours after treatment and controlled re-infestations for up to 30 days (efficacy >96.20%, p<0.05). The egg laying inhibition was over 92.3% for up to 29 days (p<0.05). The adult emergence inhibition remained 100% during 8 weeks after treatment and was 99.8% nine weeks after treatment (p<0.001).

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.

Lumpy skin disease in Ethiopia: seroprevalence study across different agro-climate zones.

A cross-sectional study was conducted to estimate the seroprevalence of lumpy skin disease (LSD) in the different agro-climatic zones prevailing in Ethiopia. A total of 2368 serum samples were collected from 42 kebeles located in 15 districts and tested using indirect fluorescent antibody test (IFAT) and virus neutralization test (VNT). The herd and animal true LSD serological prevalence were estimated in each agro-climate zone using a Bayesian model. The intra-cluster correlation coefficient (ICC) was evaluated using a random-effect model. According to the serological prevalence estimations, LSD affected differently the three agro-climatic zones considered. Herd level seroprevalence was higher in the midland agro-climate zone 64% (95% CI: 53-74) as compared to the highland 26% (95% CI: 17-36) and the lowland 50% (95% CI: 40-60) agro-climates. Animal level seroprevalence in infected herds was also higher in the midland agro-climate zone 31% (95% CI: 24-40) than in the highland and lowland zones (24% (95% CI: 18-31) and 23% (95% CI: 18-29), respectively). Higher ICC value in the highland agro-climate zone implies that increased sample sizes should be particularly required for this zone in future studies to estimate LSD prevalence or incidence with a desired precision level. This seroprevalence study also suggests that the prevalence of LSD infection in Ethiopia is higher than what has been previously reported. In the light of these updated estimations, we discuss options to trigger appropriate control measures in the future.

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.