Efficacy of fipronil combined with permethrin commercial spot on (Effitix) preventing Culex pipiens from feeding on dogs.

A controlled clinical trial was carried out to assess the adulticidal and anti-feeding effectiveness of a spot-on combining fipronil and permethrin (Effitix, Virbac, Carros, France) in preventing Culex pipiens from feeding on dogs. Twelve dogs with equal sensitivity to mosquitoes were included in the study and divided into two groups of six dogs: an untreated control group and a group treated with Effitix. All dogs were challenged with 80 females C. pipiens for 90 ± 5 min on days -7, 1, 7, 14, 21, and 28 (day 0 being treatment day). The number of engorged, dead, and live mosquitoes was determined after each exposure to treated and untreated dogs. Dead mosquitoes were also counted 24 h after exposure. The anti-feeding effect of the spot-on formulation was 100, 99.5, 97.7, 98.3, and 96.7% on days 1, 7, 14, 21, and 28, respectively. The mortality effect was 66.6, 55.9, 38, 17.2, and 12.3% on days 1, 7, 14, 21, and 28, respectively. At each challenge point, the mortality and anti-feeding effects on mosquitoes were significantly different between the control and treated group (p < 0.05). The results indicate that a combination of permethrin and fipronil could be used as an effective mosquito control strategy in dogs and is therefore recommended for use in a dirofilariasis prevention program.

Experimental infections of rabbits with proliferative and latent stages of Besnoitia besnoiti.

Cattle besnoitiosis due to Besnoitia besnoiti is spreading across Europe and is responsible for severe economic losses in newly infected herds. Experimentally speaking, rabbits have been found to be susceptible to this parasite. The adaptation of B. besnoiti to rabbits may offer a new, easier and cheaper model of investigation for this disease. This study compared the virulence between tachyzoites and bradyzoites of B. besnoiti in rabbits. Eighteen New Zealand rabbits were allocated into three groups of six animals each. The rabbits from the control (group C), "tachyzoite" (group T) and "bradyzoite" (group B) groups were subcutaneously injected in the right flank with 66 µg of ovalbumin, 6.10(6) tachyzoites (125th passage on Vero cells) and 6.10(6) bradyzoites (collected from a natural infected cow) of B. besnoiti, respectively. Clinical follow-up and blood sampling for serological survey and qPCR were performed during 10 weeks until euthanasia. Molecular and immunohistochemistry examination was achieved on 25 samples of tissue per rabbit. Seroconversion occurred in group T without any clinical signs. Rabbits of group B exhibited a febrile condition (temperature above 40 °C from day 8 to day 11 following injection) with positive qPCR in blood. Cysts of B. besnoiti were found on skin samples and organs of rabbits from group B in tissue explored with threshold cycle (Ct) values below 30. These results suggest a higher virulence of bradyzoites in rabbits than Vero cell-cultivated tachyzoites. The proposed model could be used to assess the in vivo effectiveness of vaccine or drugs against cattle besnoitiosis.

The efficacy of a selamectin (Stronghold ®) spot on treatment in the prevention of Bartonella henselae transmission by Ctenocephalides felis in cats, using a new high-challenge model.

Bartonella henselae is the causative agent of cat scratch disease in humans, which is recognized as an emerging zoonotic disease. Ctenocephalides felis is the main vector, and transmission of B. henselae infection between cats and humans occurs mainly through infected flea feces. Control of feline infestation with this arthropod vector therefore provides an important strategy for the prevention of infection of both humans and cats. In the present study, a new challenge model is used to evaluate the efficacy of selamectin (Stronghold(®) spot on) in the prevention of B. henselae transmission by C. felis. In this new challenge model, domestic cats were infected by direct application of B. henselae-positive fleas. The fleas used for infestation were infected by feeding on blood that contained in vitro-cultured B. henselae. The direct application of the fleas to the animals and the use of different B. henselae strains ensured a high and consistent challenge. Two groups of six cats were randomly allocated on pre-treatment flea counts to either control (untreated cats) or the selamectin-treated group with one pipette per cat according to the label instruction. Stronghold (selamectin 6 % spot on solution) was administered on days 0 and 32. On days 3, 10, 19, 25, and 31, each cat was infested by direct application of 20 fleas that fed on blood inoculated with B. henselae. Polymerase chain reaction (PCR) on pooled fleas confirmed that the fleas were infected. Blood samples were collected from each cat on days -3 (prior to flea infestation and treatment), 9, 17, 24, 30, 37, and 44 and assayed for B. henselae antibodies using an indirect immunofluorescence (IFA), for the presence of bacteria by bacterial culture and for B. henselae DNA presence by PCR. Cats were also assessed on a daily basis for general health. There were no abnormal health observations during the study and none of the animals required concomitant treatment. None of the cats displayed any clinical signs of bartonellosis during the study. In the untreated group, all cats became bacteremic within 17 to 44 days. None of the selamectin-treated cats became positive during the study. It was concluded that Stronghold(®) spot on administered to cats was efficacious in the prevention of the transmission of B. henselae by fleas to cats in a high-challenge model.

Differential needs of zinc in the CA3 area of dorsal hippocampus for the consolidation of contextual fear and spatial memories.

One peculiarity of the hippocampal CA3 mossy fiber terminals is the co-release of zinc and glutamate upon synaptic transmission. How these two players act on hippocampal-dependent memories is still unclear. To decipher their respective involvement in memory consolidation, a pharmacological approach was chosen. Using two hippocampal-dependent behavioral paradigms (water maze and contextual fear conditioning) we now report that glutamate at CA3 synapses is necessary and sufficient for the spatial learning consolidation process, whereas glutamate and zinc released by mossy fibers are both mandatory and exert cumulative effects on contextual fear consolidation, a form of learning with a strong emotional component.

Feline leishmaniosis in the Mediterranean Basin: a multicenter study.

BACKGROUND: Cats are now recognized as competent hosts for Leishmania infantum and a blood source for sand fly vectors. Although canine leishmaniosis (CanL) is endemic in Mediterranean Basin countries, large-scale epidemiological studies are lacking for feline leishmaniosis (FeL). This study aimed to assess the prevalence of L. infantum infections, associated risk factors, clinical signs, and clinicopathological abnormalities in domestic cat populations from six Mediterranean Basin countries. METHODS: From 2019 to 2022, blood and serum samples of cats (n = 2067) living in Italy (n = 300), Greece (n = 297), Portugal (n = 295), France (n = 231), Israel (n = 313), and Spain (n = 631) were collected along with animal data (i.e., age, sex, breed, housing conditions, and geographical origin), clinical signs, and laboratory blood test parameters. Cats were grouped according to their age as kittens (up to 1 year), young (older than 1 and younger than 7 years), mature (between 7 and 10 years), and senior (older than 10 years). Serum samples were tested for L. infantum by immunofluorescence antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA), and blood samples of seropositive cats were tested for L. infantum kinetoplast deoxyribonucleic acid (kDNA). Viral infection by feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) was molecularly addressed in all cats enrolled. Statistical analysis was performed to evaluate the association between the risk of L. infantum infection and independent variables, and among co-infection of L. infantum with FIV and/or FeLV, clinical signs, and clinicopathological abnormalities. RESULTS: Overall, 17.3% (358/2067) of cats scored positive for L. infantum by serological tests. Specifically, 24.7% were from Portugal, 23.2% from Greece, 16.6% from Israel, 15% from Spain, 13.3% from France, and 12.6% from Italy. Leishmania infantum DNA was detected in 15 seropositive animals. Housing condition and FIV infection proved to be risk factors for FeL. Leishmania seropositivity was significantly associated with weight loss, lymphadenomegaly, gingivostomatitis, and oral ulcers, as well as with reduced albumin and albumin/globulin ratio, increased total globulins and total proteins, leukocytosis, and thrombocytosis. CONCLUSIONS: This study provides, for the first time, a large-scale epidemiological survey on FeL and its clinical presentation, revealing that L. infantum circulates among domestic cats, especially shelter/free-roaming and FIV-infected animals, living in CanL endemic countries of the Mediterranean Basin.

Fleas of wild carnivores in Romania: diversity, distribution, and host-associations.

BACKGROUND: Fleas are important hematophagous insects, infesting mammals and birds with a worldwide distribution. Fleas of medical importance have been reported from various carnivores worldwide, such as felids, canids, or mustelids. Romania hosts a wide carnivore diversity, but very little is known about flea species that parasitize these animals in Romania. This study aimed to provide a better understanding of the fleas' diversity and their distribution in a relatively large and diverse number of wild carnivore hosts from Romania. METHODS: From 2013 to 2021, 282 carcasses of wild carnivores from different locations in Romania were collected and examined for the presence of ectoparasites. All collected fleas were morphologically identified using specific keys and descriptions. An analysis of the co-occurrence networks was performed. RESULTS: A total of 11 flea species were identified: Pulex irritans (41.09%), Paraceras melis (20.11%), Ctenocephalides felis (7.33%), Ctenocephalides canis (7.83%), Monopsyllus sciurorum (11.11%), Chaetopsylla trichosa (21.96%), Chaetopsylla homoea (5.5%), Chaetopsylla tuberculaticeps (100%), Chaetopsylla rothschildi (13.33%), Chaetopsylla sp. (14.34%), Chaetopsylla globiceps (5.12%), Echidnophaga gallinacea (10%). The statistical analyses showed a significant difference between the infestation of Martes foina with females being more frequently infected than males (66% versus 33%). Paraceras melis infesting Meles meles had a significantly higher prevalence in female badgers than in males (× 2 = 7.7977, P < 0.01) and higher intensities of infestations in males than in females (t = 1.871, P < 0.05). CONCLUSIONS: This is the first large-scale study investigating the distribution and diversity of flea species infesting wild carnivores in Romania. Three flea species were identified for the first time in Romania (E. gallinacea, C. homoea, and C. tuberculaticeps).

Assessment of persistence of Bartonella henselae in Ctenocephalides felis.

Bartonella henselae (Rhizobiales: Bartonellaceae) is a Gram-negative fastidious bacterium of veterinary and zoonotic importance. The cat flea Ctenocephalides felis (Siphonaptera: Pulicidae) is the main recognized vector of B. henselae, and transmission among cats and humans occurs mainly through infected flea feces. The present study documents the use of a quantitative molecular approach to follow the daily kinetics of B. henselae within the cat flea and its excreted feces after exposure to infected blood for 48 h in an artificial membrane system. B. henselae DNA was detected in both fleas and feces for the entire life span of the fleas (i.e., 12 days) starting from 24 h after initiation of the blood meal.

Systematic review of hematophagous arthropods present in cattle in France.

The arrival of pathogens, whether zoonotic or not, can have a lasting effect on commercial livestock farms, with dramatic health, social and economic consequences. However, available data concerning the arthropod vectors present and circulating on livestock farms in France are still very imprecise, fragmentary, and scattered. In this context, we conducted a systematic review of the hematophagous arthropod species recorded on different types of cattle farms in mainland France (including Corsica). The used vector "groups" studied were biting flies, biting midges, black flies, fleas, horse flies, lice, louse flies, mosquitoes, sand flies, and ticks. A large number of documents were selected (N = 9,225), read (N = 1,047) and analyzed (N = 290), allowing us to provide distribution and abundance maps of different species of medical and veterinary interest according to literature data. Despite the large number of documents collected and analyzed, there are few data provided on cattle farm characteristics. Moreover, data on all arthropod groups lack numerical detail and are based on limited data in time and/or space. Therefore, they are not generalizable nor comparable. There is still little information on many vectors (and their pathogens) and still many unknowns for most studied groups. It appears necessary to provide new, updated and standardized data, collected in different geographical and climatological areas. Finally, this work highlights the lack of entomologists, funding, training and government support, leading to an increased risk of uncontrolled disease emergence in cattle herds.

Title: Revue systématique des arthropodes hématophages présents chez les bovins en France. Abstract: L'arrivée d'agents pathogènes, même non zoonotiques, peut affecter durablement les élevages commerciaux, avec des conséquences sanitaires, sociales et économiques dramatiques. Cependant, les données disponibles concernant les arthropodes vecteurs présents et circulants dans les élevages français sont encore très imprécises, fragmentaires et dispersées. Dans ce contexte, nous avons réalisé une revue systématique des espèces d'arthropodes hématophages présentes dans différents types d'élevages bovins en France métropolitaine (Corse comprise). Les "groupes" de vecteurs retenus et étudiés étaient les mouches piqueuses, les culicoïdes, les simulies, les puces, les taons, les poux, les hippobosques, les moustiques, les phlébotomes et les tiques. Un grand nombre de documents ont été sélectionnés (N = 9 225), lus (N = 1 047) et analysés (N = 290), ce qui nous a permis de fournir des cartes de distribution et d'abondance des différentes espèces d'intérêt médical et vétérinaire en fonction des données de la littérature. Malgré le grand nombre de documents collectés et analysés, peu de données sont fournies sur les caractéristiques des élevages bovins. En outre, les données sur tous les groupes d'arthropodes manquent de détails numériques et sont basées sur des informations limitées dans le temps et/ou l'espace. Elles ne sont donc ni généralisables ni comparables. Il existe encore peu d'informations sur de nombreux vecteurs (et leurs agents pathogènes) et de multiples inconnues subsistent pour les groupes les plus étudiés. Il apparaît nécessaire de collecter de nouvelles données, actualisées et standardisées dans différentes zones géographiques et climatiques. Enfin, ce travail met en évidence le manque d'entomologistes, de financements, de formations et de soutien gouvernemental, ce qui accroît le risque d'émergence non contrôlée de maladies dans les troupeaux de bovins.

Hepatozoon species infecting domestic cats from countries of the Mediterranean basin.

Tick-borne diseases (TBDs) are caused by pathogens of human and veterinary concern representing a major public health issue worldwide. Although feline medicine has progressed much in the recent decades, data on feline TBDs (FeTBDs) remain scant. Therefore, this study aimed to assess the prevalence of apicomplexan parasite infections, associated risk factors and clinical-hematological abnormalities in domestic feline populations from countries of the Mediterranean basin. Blood and serum samples from cats (n = 600) living in France, Greece, Israel, Italy, Portugal and Spain were collected along with animal data (i.e., age, sex, breed, housing conditions and geographical origin), clinical signs and laboratory blood test parameters. Cats were grouped according to their age as kitten (up to one year), young (between one and six years), mature (between seven and ten years) and senior (older than ten years). Blood samples were tested for Hepatozoon spp. and piroplasmids by conventional PCR targeting 18S rRNA gene. The overall prevalence of Hepatozoon spp. infection was 14.5%, being significantly higher in cats from Greece (30%) and Portugal (23%), followed by Spain (15%), Israel (15%) and France (4%). Cats from Italy scored negative. Hepatozoon felis was identified in 86 animals, with three different sequence types and H. silvestris was detected in one shelter cat from Portugal. No piroplasmid DNA was amplified. The risk of Hepatozoon spp. infection was related to feline geographical provenience, housing condition and age. No statistical correlation was reported with any clinical signs, while increased alanine aminotransferase (ALT) activity was the only laboratory abnormality significantly associated (p = 0.03) with the infection. Data suggest a high circulation of H. felis, and only occasionally of H. silvestris, within domestic feline populations in the Mediterranean basin, mainly in shelter or free roaming and young cats with asymptomatic or subclinical infection.

Therapy and Prevention for Human Toxocariasis.

For the last four decades, knowledge about human toxocariasis with regard to its epidemiology, pathophysiology, clinical spectrum, and imaging or laboratory diagnosis has substantially progressed. Knowledge about specific therapy with anthelmintics has lagged behind. To date, only four drugs are registered for human use, and their efficacy has rarely been assessed in prospective controlled trials. It is likely that the repurposing of potent anthelmintics from veterinary medicine will improve this situation. Due to its wide availability and a lack of major side effects during short regimens, albendazole has become the drug of choice. However, its efficacy should be more precisely assessed. The role of anthelmintics in the treatment of neurological or ocular toxocariasis remains to be clarified. Prophylactic measures in humans or companion animals are efficient and represent first-line treatments for the control of this zoonosis. Unfortunately, their implementation in areas or countries where toxocariasis epidemiology is driven by poverty is quite difficult or unrealistic.

Prevalence of major digestive and respiratory helminths in dogs and cats in France: results of a multicenter study.

BACKGROUND: The local distribution of helminths in dogs and cats and the evaluation of risk of contamination represent an important challenge for veterinarians due to their effects on animal health and their potential zoonotic risk. The overall goal of this study was to estimate the prevalence of the digestive and respiratory helminths infecting client-owned dogs and cats in France. METHODS: Faecal samples were collected from 414 pet dogs and 425 pet cats at 20 study sites during 2017-2018 and analysed by coproscopy. The samples included specimens collected from animals of both genders and various breeds and ages from a variety of living environments, and with different lifestyles and feeding regimes. Associations between parasitic infection and qualitative factors were explored. RESULTS: Overall, 125 (14.9%) samples (15.2% in dogs and 14.6% in cats) were positive for at least one of the species of helminths identified. Infection rates were highest for Toxocara canis and Toxocara cati (8.5% and 11.3%, respectively), while Toxascaris leonina was found only in one cat (0.2%). The apparent prevalence of Ancylostoma caninum and Uncinaria stenocephala in dogs was 1.7% and 4.3%, respectively. No hookworms were found in cats. Whipworms (Trichuris vulpis) were identified in 2.7% of the dogs. Tapeworms (Dipylidium caninum and Taeniidae) were rarely found (< 1% in dogs and < 3% in cats). The prevalence of Angiostrongylus vasorum Crenosoma vulpis, and Strongyloides stercoralis in dogs, Aelurostrongylus abstrusus in cats and Eucoleus spp. / Capillaria spp. in both dogs and cats was < 1%. Significantly higher fecal parasite emission rates were identified in young individuals, in animals with outdoor access, in animals living in the countryside and in intact animals (especially in cats). In addition, cats not fed exclusively with commercial diets and living with other animals (dogs and/or cats) were at higher risk for parasites. For dogs, hunting/herding and walking off-leash were found to be additional risk factors. Furthermore, pets with no reported history of deworming or dewormed > 1 year before the study were positive for parasites significantly more often than pets dewormed < 1 year before study participation. CONCLUSIONS: The overall prevalence of helminths (some of which are zoonotic), the risk factors and the reportedly low deworming frequencies identified in this study (20.5% animals having never been dewormed and only 26.4% dewormed ≥ 3 times/year) illustrate the need for improving pet owners' adherence to anthelmintic guidelines in France.

An expert opinion assessment of blood-feeding arthropods based on their capacity to transmit African swine fever virus in Metropolitan France.

To deal with the limited literature data on the vectorial capacity of blood-feeding arthropods (BFAs) and their role in the transmission of African swine fever virus (ASFV) in Metropolitan France, a dedicated working group of the French Agency for Food, Environmental and Occupational Health & Safety performed an expert knowledge elicitation. In total, 15 different BFAs were selected as potential vectors by the ad hoc working group involved. Ten criteria were considered to define the vectorial capacity: vectorial competence, current abundance, expected temporal abundance, spatial distribution, longevity, biting rate, active dispersal capacity, trophic preferences for Suidae, probability of contact with domestic pigs and probability of contact with wild boar. Fourteen experts participated to the elicitation. For each BFA, experts proposed a score (between 0 and 3) for each of the above criteria with an index of uncertainty (between 1 and 4). Overall, all experts gave a weight for all criteria (by distributing 100 marbles). A global weighted sum of score per BFA was calculated permitting to rank the different BFAs in decreasing order. Finally, a regression tree analysis was used to group those BFAs with comparable likelihood to play a role in ASF transmission. Out of the ten considered criteria, the experts indicated vectorial competence, abundance and biting rate as the most important criteria. In the context of Metropolitan France, the stable fly (Stomoxys calcitrans) was ranked as the most probable BFA to be a vector of ASFV, followed by lice (Haematopinus suis), mosquitoes (Aedes, Culex and Anopheles), Culicoides and Tabanidea. Since scientific knowledge on their vectorial competence for ASF is scarce and associated uncertainty on expert elicitation moderate to high, more studies are however requested to investigate the potential vector role of these BFAs could have in ASFV spread, starting with Stomoxys calcitrans.

Mechanical transmission of African swine fever virus by Stomoxys calcitrans: Insights from a mechanistic model.

African swine fever (ASF) represents a global threat with huge economic consequences for the swine industry. Even though direct contact is likely to be the main transmission route from infected to susceptible hosts, recent epidemiological investigations have raised questions regarding the role of haematophagous arthropods, in particular the stable fly (Stomoxys calcitrans). In this study, we developed a mechanistic vector-borne transmission model for ASF virus (ASFV) within an outdoor domestic pig farm in order to assess the relative contribution of stable flies to the spread of the virus. The model was fitted to the ecology of the vector, its blood-feeding behaviour and pig-to-pig transmission dynamic. Model outputs suggested that in a context of low abundance (<5 flies per pig), stable flies would play a minor role in the spread of ASFV, as they are expected to be responsible for around 10% of transmission events. However, with abundances of 20 and 50 stable flies per pig, the vector-borne transmission would likely be responsible for almost 30% and 50% of transmission events, respectively. In these situations, time to reach a pig mortality of 10% would be reduced by around 26% and 40%, respectively. The sensitivity analysis emphasized that the expected relative contribution of stable flies was strongly dependent on the volume of blood they regurgitated and the infectious dose for pigs. This study identified crucial knowledge gaps that need to be filled in order to assess more precisely the potential contribution of stable flies to the spread of ASFV, including a quantitative description of the populations of haematophagous arthropods that could be found in pig farms, a better understanding of blood-feeding behaviours of stable flies and the quantification of the probability that stable flies partially fed with infectious blood transmit the virus to a susceptible pig during a subsequent blood-feeding attempt.

Attractiveness and Specificity of Different Polyethylene Blue Screens on Stomoxys calcitrans (Diptera: Muscidae).

Stomoxys calcitrans is considered as a major pest of livestock worldwide. Insecticides have been extensively used to control this pest but resistance to these chemical compounds is now reported in many countries. Therefore, a more sustainable and efficient control is needed. Seven different types of blue screens, with reflectances around 460 nm, were tested during summer 2016 in southwestern France to evaluate their attractiveness and their specificity for stable flies. Height of the screen and orientation (east or west) of a blue screen were also considered. High levels of S. calcitrans captures were recorded during this study (from 141 to 7301 individuals per blue screen and per day) whereas the numbers of tabanids and pollinator insects remained extremely low (less than 10 individuals per screen and per day). No significant difference in attractiveness has been shown between the different types of blue screens. The lower half of the blue screens caught significantly more stable flies (70%) than the higher half (30%). The "east" side of the screen attracted 60% of stable flies but this was not significantly different from the west side. These results are highlighting the interest in these blue polyethylene screens for controlling stable flies in cattle farms, in comparison with more expensive blue fabrics.

Putative Role of Arthropod Vectors in African Swine Fever Virus Transmission in Relation to Their Bio-Ecological Properties.

African swine fever (ASF) is one of the most important diseases in Suidae due to its significant health and socioeconomic consequences and represents a major threat to the European pig industry, especially in the absence of any available treatment or vaccine. In fact, with its high mortality rate and the subsequent trade restrictions imposed on affected countries, ASF can dramatically disrupt the pig industry in afflicted countries. In September 2018, ASF was unexpectedly identified in wild boars from southern Belgium in the province of Luxembourg, not far from the Franco-Belgian border. The French authorities rapidly commissioned an expert opinion on the risk of ASF introduction and dissemination into metropolitan France. In Europe, the main transmission routes of the virus comprise direct contact between infected and susceptible animals and indirect transmission through contaminated material or feed. However, the seasonality of the disease in some pig farms in Baltic countries, including outbreaks in farms with high biosecurity levels, have led to questions on the possible involvement of arthropods in the transmission of the virus. This review explores the current body of knowledge on the most common arthropod families present in metropolitan France. We examine their potential role in spreading ASF-by active biological or mechanical transmission or by passive transport or ingestion-in relation to their bio-ecological properties. It also highlights the existence of significant gaps in our knowledge on vector ecology in domestic and wild boar environments and in vector competence for ASFV transmission. Filling these gaps is essential to further understanding ASF transmission in order to thus implement appropriate management measures.

Evaluation of the anti-feeding and insecticidal effects of a topically administered combination of imidacloprid and permethrin (Advantix®) against Phlebotomus (Larroussius) perniciosus (Newstead, 1911) in dogs following monthly administration.

BACKGROUND: Two laboratory experiments (Studies 1 and 2) were conducted to confirm the efficacy of an imidacloprid and permethrin combination (Advantix® Spot-on, Bayer) to repel and kill Phlebotomus (Larroussius) perniciosus sand flies when applied once a month topically to dogs. METHODS: Both studies compared dogs treated with a combination containing 100 mg/ml imidacloprid + 500 mg/ml permethrin (Advantix® Spot-on, Bayer) to placebo treated dogs. The treatments were applied topically on Day -28 (Study 2) and Day 0 (Studies 1 and 2). Sand fly exposures with 80 unfed females were performed before the first treatment for allocation purposes and post-treatment on study days (SDs) 1, 7, 14, 21 and 28 (following first or second monthly treatment for Studies 1 and 2, respectively). After 60 min, sand flies were assessed for mortality and engorgement status. RESULTS: Repellent evaluation (anti-feeding effect) on all days post-infestation showed efficacies that ranged between 88.1-99.3% during the first month and 92.2-98.9% during the second. Analyses of the comparison of fed sand fly counts for each treatment group resulted in a highly significant reduction (P < 0.0001) at all post-infestation time points for those dogs treated with Advantix®. A significant (P < 0.0001 for all time points) insecticidal effect was equally demonstrated. No treatment related adverse events were observed during the study. CONCLUSIONS: In the present studies Advantix® Spot-on demonstrated to be safe and to provide excellent four-week sand fly (P. perniciosus) repellency of ≥88.1% and ≥92.2% after a first and second monthly treatment, respectively. A significant insecticidal effect was also observed.

First evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium.

BACKGROUND: Besnoitiosis is caused by different species of intracellular protozoan parasites belonging to the family Sarcocystidae and affecting multiple host species worldwide. Including B. besnoiti, ten species are described infecting animals. Among ungulates, Besnoitia bennetti infects horses, donkeys and zebras and was described in Africa and in the USA where donkey besnoitiosis is considered as an emerging disease. CASE PRESENTATION: A two-year-old male donkey was purchased in May 2016 in poor body condition (cachexia, alopetic areas and pruritus mainly on neck and head) by the present owner in Le Roeulx (Belgium) from a milk producing donkey farm in Frasnes-lez-Buissenal (Belgium). Shortly after its purchase and shearing, the donkey presented with crusts, hyperkeratosis (both flanks and neck) anorexia and cachexia. A treatment with phoxim was given with no improvement. A cutaneous biopsy of hyperkeratotic skin was performed in July. It showed a perivascular eosinophilic infiltrate with a large thick walled cyst located in the dermis containing numerous bradyzoites. This was highly suggestive of besnoitiosis. Several skin biopsy samples were obtained for qPCR analysis and confirmed the presence of Besnoitia spp. DNA. Further laboratory diagnosis tests were performed (western blot and rDNA sequencing) confirming Besnoitia bennetti aetiology for the male. For the female, the punch-biopsy, haematology and qPCR were negatives but the western blot showed the presence of antibodies directed to Besnoitia spp. Further clinical examination performed in August highlighted scleral pinhead sized cysts (pearl) in the right eye and between nares. Another ten-year-old female donkey purchased in France and sharing the same accommodation showed a good clinical condition, but a thorough clinical examination showed the presence of numerous cysts on the inner face of upper labial mucosa. A daily treatment based on sulfamethaxzole and trimethoprim (Emdotrim 60% Mix®, 30 mg/kg) was given orally and some improvement was noticed. CONCLUSION: This is the first evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium.

Repellent and insecticidal efficacy of a combination of dinotefuran, pyriproxyfen and permethrin (Vectra® 3D) against Culex pipiens in dogs.

Culex pipiens is an important vector of pathogens of substantial medical and veterinary importance such as Dirofilaria immitis and Dirofilaria repens or the West Nile Virus. The control of these mosquitoes is therefore essential to control the transmission of mosquito-borne agents to humans and animals. A combination of dinotefuran, permethrin and pyriproxyfen (Vectra® 3D) has already shown its efficacy against Aedes aegypti. The aim of this study was to confirm the efficacy of this combination in repelling and killing another species of mosquito, Culex pipiens, after a single topical application to dogs. Twelve adult Beagle dogs with an equal receptivity to mosquitoes were included in the study and divided in two groups of six dogs: an untreated control group and a group treated with a combination containing 54 mg/mL dinotefuran + 4.84 mg/mL pyriproxyfen + 397 mg/mL permethrin (Vectra® 3D). All dogs were challenged with 80 Culex pipiens females for 90 ± 5 min on Days - 28, 1, 7, 14, 21 and 28. The treatment was applied once topically on Day 0. Count and engorgement determination of dead and live mosquitoes were performed after each exposure to treated and untreated dogs. Compared to control dogs, the spot-on formulation provided a repellent efficacy (anti-feeding effect) against mosquitoes of 98.9%, 98.8%, 98.6%, 96.7% and 97.9% on Days 1, 7, 14, 21 and 28 respectively. There was a significant difference (p ≤ 0.05) between the treated and controlled groups on every assessment day. The insecticidal efficacy on treated dogs at 90 min was 34.7%, 50.3%, 39.7%, 22.8% and 11.4% on Days 1, 7, 14, 21 and 28 respectively. There was a significant difference between the treated and controlled groups for live mosquitoes for all assessment days (p < 0.05). A single topical application of a combination of dinotefuran, permethrin and pyriproxyfen showed a significant repellent effect (i.e. > 96%) against Culex pipiens which lasted for 28 days. The results suggest that the Vectra® 3D spot-on solution could be used as an effective mosquito control strategy in dogs and is therefore recommended for use in a dirofilariosis prevention programme.

Sustained speed of kill and repellency of a novel combination of fipronil and permethrin against Ctenocephalides canis flea infestations in dogs.

BACKGROUND: Ctenocephalides canis is a major flea species in dogs in several European countries. The new topical combination of fipronil and permethrin (Frontline Tri-Act/Frontect, Merial) has been developed to control fleas, ticks, mosquitoes, sandflies and biting flies on dogs. Considering the repellent and insecticidal effects of permethrin and the insecticidal effect of fipronil, the efficacy of the combination against fleas including C. canis was expected to be rapid. The study was conducted to measure the 1-hour, 6-hour and 24-hour efficacy, as well as the repellent activity, of the fipronil-permethrin combination on treated versus untreated dogs. METHODS: 12 Beagle dogs were randomly allocated to one of two groups based on pre-treatment live flea counts. Dogs in Group 1 remained untreated whereas dogs in Group 2 were treated once on Day 0. Each dog was infested with 100 unfed adult C. canis on Days 2, 7, 14, 21 and 28. Dogs were combed for fleas 1 and 6 h after each infestation. Following this examination, fleas remaining on the liner at the bottom of each cage were collected and counted. All live fleas were placed back on each dog after the 1- and 6-hour counts. A comb-count was performed at 24 h post infestation on all dogs. RESULTS: Treated dogs had significantly (p ≤ 0.01) lower flea counts than untreated dogs at every time point. The percent efficacy was ≥99.1% at 6 and 24 h after each weekly challenge up to the month. The 1-hour counts also showed good efficacy of 96.5%, 98.9%, 92.0%, 70.2% and 55.7% on Days 2, 7, 14, 21 and 28, respectively. The repellent efficacy, assessed on the liners at 1 h, was 86.5%, 94.9%, 79.5%, 58.4% and 43.9% on Days 2, 7, 14, 21 and 28, respectively. CONCLUSIONS: This study demonstrates the beneficial effect of the fipronil and permethrin combination against C. canis, providing both a repellent and insecticidal effect as early as 1 h post infestation, and >99.1% efficacy calculated at 6 h during a month.

Intravenous lipid emulsion therapy in 20 cats accidentally overdosed with ivermectin.

OBJECTIVE: To describe the outcome of 20 cats treated with intravenous lipid emulsion (IVLE) after an accidental parenteral ivermectin overdose. CASE SERIES SUMMARY: Twenty adult cats presented after receiving a 4 mg/kg accidental subcutaneous overdose of ivermectin. After admission, two IVLE treatments were initiated in asymptomatic cats: a single bolus (1.5 mL/kg; n = 16) versus a bolus followed by a 30-minute constant rate infusion (0.25 mL/kg/min; n = 4). Six out of the 16 cats that received only the single bolus developed clinical signs of ivermectin intoxication. Based on the severity of the clinical signs and their duration (approximately 48 hours), these 6 cats were retrospectively considered either moderately (n = 3) or severely (n = 3) intoxicated by ivermectin. Cats with a low body condition score (BCS) had more severe signs of ivermectin toxicity. Additional IVLE was administered until clinical resolution was complete. Median (min to max) cumulative dose of IVLE per cat was 4.5 (3.0-4.5) mL/kg for 36 (12-36) hours and 19.5 (7.5-37.5) mL/kg for 96 (72-168) hours for moderately and severely intoxicated cats, respectively. NEW OR UNIQUE INFORMATION PROVIDED: Our series describes the treatment of accidental ivermectin parenteral overdose in 20 cats with early initiation of IVLE therapy. An early bolus followed by a 30-minute constant rate infusion of IVLE appeared to mitigate the signs of ivermectin toxicosis in cats compared to a single treatment bolus. Our observations also suggest that cats with a low BCS given only a bolus of IVLE treatment were more likely to develop signs of ivermectin intoxication and require a greater amount of IVLE for the resolution of clinical signs. Based on our observations, BCS appears to influence the severity of ivermectin toxicity with a low BCS being associated with more severe signs of ivermectin toxicity.