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

The occurrence of the nematode Angiostrongylus vasorum, also known as the French heartworm, is increasingly being reported from various European countries. The adults of this parasite species live in the pulmonary arteries and right cardiac ventricle of wild canids and domestic dogs. Larval stages and eggs in the lungs induce inflammatory verminous pneumonia, causing severe respiratory disease in dogs. Furthermore, haematological and neurological signs and even death may occur. In Hungary, A. vasorum has been identified in red foxes, golden jackals and in two dogs and some slugs. In this first large-scale survey, 1247 sera from pet dogs were collected and tested by an ELISA for the detection of circulating antigen of A. vasorum and by a separate ELISA to detect specific antibodies against the parasite. A total of 1.36% (n = 17, 95 % confidence intervals, CI: 0.80 - 2.17 %) of the animals were positive in both ELISAs, while 1.76 % (n = 22, CI: 1.11 - 2.66 %) of the tested dogs were antigen-positive only and 2.73 % (n = 34, CI: 1.90 - 3.79 %) were positive for specific antibodies only. Regions with antigen- and antibody-positive animals overlapped and were distributed over nearly the whole sampled areas of the country. A considerable number of cases was observed in Budapest and also in the southern part of the country bordering Croatia, while in the most eastern part bordering Ukraine no positive samples were detected. These results confirm the endemic occurrence of A. vasorum in dogs originating from different parts of Hungary and the significant advantages of A. vasorum serology in epidemiological studies.

Cats infected with the metastrongylid nematode Aelurostrongylus abstrusus may show clinical signs ranging from mild to severe respiratory disease or remain unobserved, despite damages present in the lung tissue. This study aimed to determine the seroprevalence and distribution of A. abstrusus in cats by testing serum samples from all over Germany to identify potential risk areas and strengthen disease awareness accordingly. Sera of 2998 cats were screened for the presence of antibodies against A. abstrusus by ELISA, and the data were evaluated by a geographic information system to visualise the regional distribution of the analysed samples. Overall, 12.0% of the samples tested positive (361/2998 cats, 95% confidence interval: 10.9–13.3%). Seropositive cats were identified throughout the country, suggesting that all cats in Germany with outdoor access are at risk of A. abstrusus infection and that the infection is overall underdiagnosed. Increased testing for A. abstrusus infection would allow earlier detection of infected animals, hence improving the life quality and health of cats and preventing potential death under anaesthesia.

Background Troglostrongylus brevior is a lungworm of wild felids that recently has been recognized as agent of severe respiratory disease in domestic cats in Mediterranean and Balkan countries. Nevertheless, the information on treatment options for feline troglostrongylosis is still poor. The aim of this pilot field trial was to evaluate the efficacy of the spot-on formulation containing 1% w/v moxidectin and 10% w/v imidacloprid (Advocate® spot-on solution for cats, Bayer Animal Health GmbH) in the treatment of T. brevior infection in naturally infected cats in Greece. Methods The trial was a negative control, multicentre, clinical efficacy study conducted according to the standards of Good Scientific Practice (GSP). Sixteen cats in two study sites, naturally infected with T. brevior, were allocated to an untreated control group (G1, n = 8) or a treatment group (G2, n = 8), according to a randomization list. Animals assigned to G2 were treated with Advocate® for cats on days 0 and 28 at the recommended dose rate and animals assigned to G1 received a rescue treatment with the same product on days 56 and 84. Efficacy was assessed on days 28 and 56 in G2 and on days 84 and 112 in G1 by faecal larval counts. The primary efficacy criterion was the absence of T. brevior first-stage larvae (L1) following treatment. Other efficacy parameters were the quantitative comparison of L1 presence before (baseline) and after one or two treatments in both groups. Results All G2 cats were negative for T. brevior L1 at the first post-treatment evaluation (100% efficacy) while G1 cats were persistently shedding L1. The difference of the mean number of L1 per gram between G2 and G1 was statistically significant (P T. brevior L1 at the first post-rescue-treatment evaluation. Therefore, treatment efficacy at study completion was 100% in both groups in terms of stopping the L1 shedding in the faeces of the animals. No adverse effects were observed during the study. Conclusions These results indicate that Advocate® spot-on solution for cats represents an option for treating cats naturally infected with T. brevior.

Simple Summary Angiostrongylus vasorum is a cardiopulmonary nematode that affects canids, residing in the pulmonary artery and right atrium/ventricle. Due to its location, the parasite will have a close interaction with the different components of the innate immune system, including endothelial cells and polymorphonuclear neutrophils (PMN). Here we evaluated the expression of adhesion molecules of canine aortic endothelial cells (CAEC), and NETs formation by co-culture of freshly isolated canine PMN with A. vasorum L3. Overall, we found distinct inter-donor variations in adhesion molecule expression among CAEC isolates. Additionally, PMN and A. vasorum co-culture induced NETs release without affecting larval viability. Abstract Due to its localization in the canine blood stream, Angiostrongylus vasorum is exposed to circulating polymorphonuclear neutrophils (PMN) and the endothelial cells of vessels. NETs release of canine PMN exposed to A. vasorum infective stages (third stage larvae, L3) and early pro-inflammatory immune reactions of primary canine aortic endothelial cells (CAEC) stimulated with A. vasorum L3-derived soluble antigens (AvAg) were analyzed. Expression profiles of the pro-inflammatory adhesion molecules ICAM-1, VCAM-1, P-selectin and E-selectin were analyzed in AvAg-stimulated CAEC. Immunofluorescence analyses demonstrated that motile A. vasorum L3 triggered different NETs phenotypes, with spread NETs (sprNETs) as the most abundant. Scanning electron microscopy confirmed that the co-culture of canine PMN with A. vasorum L3 resulted in significant larval entanglement. Distinct inter-donor variations of P-selectin, E-selectin, ICAM-1 and VCAM-1 gene transcription and protein expression were observed in CAEC isolates which might contribute to the high individual variability of pathological findings in severe canine angiostrongylosis. Even though canine NETs did not result in larval killing, the entanglement of L3 might facilitate further leukocyte attraction to their surface. Since NETs have already been documented as involved in both thrombosis and endothelium damage events, we speculate that A. vasorum-triggered NETs might play a critical role in disease outcome in vivo.

Background Onchocerca lupi and Cercopithifilaria spp. are vector-borne filarioids of dogs, which harbour skin microfilariae (mfs), the former being of zoonotic concern. Proper treatment studies using compounds with microfilaricidal activity have not been performed. Therefore, this study aimed to assess the efficacy of a commercially available spot-on formulation containing moxidectin 2.5%/imidacloprid 10% for the treatment of O. lupi or Cercopithifilaria spp. skin-dwelling mfs in naturally infected dogs. Methods Privately owned dogs (n = 393) from southern Portugal were sampled via skin biopsies to identify and count mfs in 20 µl of skin sediment. A total of 22 mfs-positive dogs were allocated to treatment group (n = 11; G1) or left untreated as a control (n = 11; G2). As a pilot investigation to test the treatment efficacy, five dogs assigned to G1 were treated four times at monthly intervals with moxidectin 2.5%/imidacloprid 10% spot-on formulation on SDs 0, 28 (± 2), 56 (± 2), and 84 (± 2). Based on the negative results for both O. lupi and/or Cercopithifilaria spp. mfs of dogs in the pilot study from SD28 onwards, the remaining six dogs in G1 were treated at SD0 and assessed only at SD28. Results Of the 393 animals sampled, 78 (19.8%) scored positive for skin-dwelling mfs. At the pilot investigation, a mean number of 19.6 mfs for O. lupi was recorded among five infected dogs whereas no mfs were detected at SD28. At SD0, the mean number of Cercopithifilaria spp. larvae was 12.6 for G1 and 8.7 for G2. The mean number of mfs for G2 was 20.09. Conclusions Results herein obtained suggest that a single treatment with moxidectin 2.5%/imidacloprid 10% spot-on formulation is efficacious against skin-dwelling mfs in dogs. The microfilaricidal effect of moxidectin could also be useful in reducing the risk of O. lupi infection for humans. Graphical Abstract

Canine angiostrongylosis by Angiostrongylus vasorum is increasingly reported in both enzootic and previously free areas. The complex pathogenesis of the disease makes the clinical workup challenging. Infected dogs show highly variable clinical pictures, characterized by subclinical to life-threatening general, cardio-respiratory, neurological and/or gastrointestinal signs. The present study reports the high variability of clinical pictures from 36 dogs across central and southern Italy that were naturally infected by A. vasorum. Of them, 23 (63.9%) presented at least one clinical sign, while 13 (36.1%) were subclinically infected and apparently healthy. Overall, 19 dogs (52.8%) showed cardiorespiratory signs, 14 (38.9%) had non-specific abnormalities, 2 (5.6%) presented coagulation disorders and 1 (2.8%) had a severe neurological condition. Importantly, four dogs presenting with clinical signs had neither cough nor dyspnea. These results underline that angiostrongylosis should be included in the differential diagnosis, even when dogs display only non-specific clinical signs. The proportion of apparently healthy dogs highlights the relevance of routine copromicroscopic and/or antigenic tests in enzootic areas to avoid the sudden onset of potentially life-threatening signs.

Canine and feline cardiorespiratory parasites are of utmost relevance in veterinary medicine. Key epizootiological information on major pet metastrongyloids, i.e., Angiostrongylus vasorum and Crenosoma vulpis infecting dogs, and Aelurostrongylus abstrusus and Troglostrongylus brevior infecting cats, is missing from Austria. This study investigated their occurrence in 1320 gastropods collected in the Austrian provinces of Styria, Burgenland, Lower Austria, and in metropolitan Vienna. Metastrongyloid larvae were microscopically detected in 25 samples, and sequence analysis confirmed the presence of metastrongyloids in nine samples, i.e., A. vasorum in one slug (Arion vulgaris) (0.07%), C. vulpis in five slugs (one Limax maximus and four A. vulgaris) (0.4%), A. abstrusus in two A. vulgaris (0.17%), and the hedgehog lungworm Crenosoma striatum was detected in one A. vulgaris. The present study confirms the enzooticity of major cardiorespiratory nematodes in Austria and that canine and feline populations are at risk of infection.

Background In three randomized, controlled laboratory efficacy studies, the efficacy in the prevention of patent infections of a topical combination of imidacloprid 10%/moxidectin 1% (Advocate® spot-on formulation for cats, Bayer Animal Health GmbH) against larval stages and immature adults of Aelurostrongylus abstrusus, as well as the treatment efficacy of a single or three monthly treatments against adult A. abstrusus, were evaluated. Methods Cats were experimentally inoculated with 300–800 third-stage larvae (L3). Each group comprised 8 animals and the treatment dose was 10 mg/kg bodyweight (bw) imidacloprid and 1 mg/kg bw moxidectin in each study. Prevention of the establishment of patent infections was evaluated by two treatments at a monthly interval at three different time points before and after challenge infection. Curative efficacy was tested by one or three treatments after the onset of patency. Worm counts at necropsy were used for efficacy calculations. Results In Study 1, the control group had a geometric mean (GM) of 28.8 adult nematodes and the single treatment group had a GM of 3.4 (efficacy 88.3%). In Study 2, the control group had a GM of 14.3, the prevention group had a GM of 0 (efficacy 100%), while the treatment group had a GM of 0.1 (efficacy 99.4%). In Study 3, the GM worm burden in the control group was 32.6 compared to 0 in all three prevention groups (efficacy 100% for all of those groups). Conclusions The monthly administration of Advocate® reliably eliminated early larval stages and thereby prevented lung damage from and patent infections with A. abstrusus in cats. Regarding treatment, a single application of Advocate® reduced the worm burden, but it did not sufficiently clear the infection. In contrast, three monthly treatments were safe and highly efficacious against A. abstrusus.

Canine heartworm disease (CHD) results from infection with Dirofilaria immitis and while it is of global concern, it is most prevalent in tropical climates where conditions support the parasite and vector life cycles. Melarsomine dihydrochloride is the sole treatment for CHD recommended by the American Heartworm Society. However, in cases where cost or access to melarsomine precludes treatment of an infected dog, therapeutic alternatives are warranted. This randomized, controlled field study evaluated the adulticidal efficacy of a combination therapeutic protocol using 10 % imidacloprid + 2.5 % moxidectin spot-on and a single 28-day course of doxycycline and compared with that of a 2-dose melarsomine dihydrochloride protocol. Of 37 naturally-infected domestic dogs with class 1, 2 or early class 3 CHD enrolled in the study, 30 were evaluated for a minimum of 12 months. Seven dogs were withdrawn due to canine ehrlichiosis, non-compliance, or wrongful inclusion. Dogs were randomly assigned to a control (CP, n = 15) or investigational (IVP, n = 15) treatment group. CP dogs received two injections of melarsomine dihydrochloride (2.5 mg/kg) 24 -hs apart and maintained on monthly ivermectin/pyrantel. IVP dogs were treated with oral doxycycline (10 mg/kg twice daily for 28 days) and topical 10 % imidacloprid + 2.5 % moxidectin once monthly for 9 months. Dogs were evaluated up to 18 months - monthly for the first 9 months, then every 3 months. Parasiticidal efficacy was based on antigen status using the IDEXX PetChek® 34 Heartworm-PF Antigen test. By month 18, antigen was not detected in any study dog except one from the IVP group. One other IVP dog was persistently antigenemic and treated with melarsomine at month 12 according to the initial study protocol. Mean antigen concentration (based on optical density) decreased more rapidly in the CP group and by month 15 was 0.11 for the IVP and 0.07 for CP groups, with equivalent median concentrations (0.04) in both groups. Conversion following heat-treatment of antigen-negative samples occurred frequently and at similar rates in both treatment groups. Based on the bias of diagnostic tests towards detection of female worms, we conclude that monthly application of 10 % imidacloprid + 2.5 % moxidectin for 9 months combined with a course of doxycycline twice daily for 28 days resulted in effective therapy against female adults in CHD. This therapeutic option may be particularly useful in cases where financial constraint or access to melarsomine precludes treatment of an infected individual. This study was supported by Bayer Animal Health.

Background Aelurostrongylus abstrusus is the most important nematode affecting the respiratory tract of cats in terms of prevalence and clinical relevance. The aim of this randomised controlled field study was to confirm the efficacy of the spot-on containing emodepside/praziquantel (Profender, Bayer Animal Health) in the treatment of aelurostrongylosis. Methods Seventeen cats with aelurostrongylosis and presenting with clinical and/or radiographic signs were included in the study. Eight cats received two biweekly doses of emodepside/praziquantel, while nine cats were allocated to a control group and received a rescue treatment at the end of the study. Clinical response was the primary outcome, while the secondary end point was the reduction of larval shedding in faeces. Results Two weeks after the first application, the cats showed a significant, though partial, recovery of clinical signs with complete clinical and parasitological resolution. The resolution of inflammatory leucogram and a significant reduction of radiographic lesions were observed two weeks after the second treatment. Red blood cells and albumin values significantly increased after eight weeks from the second application, together with the complete regression of radiographic patterns. Conclusion Two applications of this spot-on solution two weeks apart assured complete cessation of larval shedding and led to a complete clinical, clinicopathological and radiographic recovery.

The cat lungworm Aelurostrongylus abstrusus is a cause of lower respiratory tract disease worldwide. Bronchoscopy and bronchoalveolar lavage (BAL) are important tools for diagnosing respiratory diseases in cats. Therefore, the aim of the study was to investigate the usefulness of bronchoscopy and BAL in the diagnosis of A. abstrusus. Findings from bronchoscopic examination and BAL of 24 naturally infected cats were evaluated and compared with those of 12 cats with idiopathic Feline Bronchial Diseases (FBDs). Data were analyzed using Mann-Whitney or Fisher's exact tests. No significant bronchoscopic differences were detected between cats with aelurostrongylosis and FBDs in bronchial mucus, nodular lesions, and airway collapse. On the other hand, airway hyperemia, epithelial irregularities, and bronchial stenosis were observed more frequently in cats affected by FBDs than aelurostrongylosis, while bronchiectasis was found only in cats infected by A. abstrusus. Neutrophilic, eosinophilic, lymphocytic, and mixed inflammation were recorded in both groups. Bacteria or bacterial DNA was identified regardless of the presence or absence of A. abstrusus with no significant differences between groups. Larvae of A. abstrusus were cytologically detected in 5 of the 24 cats (20.8%) with aelurostrongylosis. These results indicate that, although some findings on bronchoscopic examination (i.e., bronchiectasis) can be described more frequently in cats infected by A. abstrusus, bronchial alterations and cytological findings in aelurostrongylosis are not specific unless larvae are observed and overlap with those of other feline airway diseases.

Background Angiostrongylus vasorum is a metastrongyloid nematode localized in the right heart and the pulmonary arteries of domestic dogs. The number of reports in Europe has recently increased, presumably as a consequence of a growing awareness among clinicians, animal owners and researchers, but also due to a growing incidence and territorial spread. So far, no studies have been conducted to assess the prevalence and distribution of A. vasorum in domestic dogs in Romania, and the awareness among veterinarians is limited or absent. The aim of the present study was to evaluate the countrywide seroprevalence of circulating antigens of A. vasorum and specific antibodies against A. vasorum in domestic dogs from Romania. Methods Between November 2016 and July 2017, blood was sampled from a total of 1545 domestic dogs from 23 counties of Romania. Details about their gender, age, breed, housing, use and origin were collected. All serum samples were tested for the presence of A. vasorum circulating antigens (AG) using monoclonal and polyclonal antibodies in a sandwich ELISA. Additionally, a sandwich ELISA using A. vasorum adult somatic antigen purified by monoclonal antibodies was used for specific antibody (AB) detection. Results A total of 33 dogs (2.14%, 95% CI: 1.82–3.56%) were seropositive for A. vasorum antigen or antibodies against the parasite. Three dogs were positive for antigen only (0.19%, 95% CI: 0.07–0.57%) and 30 dogs (1.94%, 95% CI: 1.36–2.76%) were positive for antibodies only. No dog was positive for both tests. The overall prevalence (AB or AG) and the AB prevalence were significantly higher in pure breed dogs compared to mixed breeds and mongrel dogs (P

Background Troglostrongylus brevior, a lungworm usually affecting wild felids, has been recently recorded in a number of cases in domestic cats, mainly in Mediterranean areas. Although feline troglostrongylosis is a severe and life-threatening disease, especially in young cats, treatment options are very limited. The present study evaluated the efficacy and safety of a spot-on formulation containing emodepside 2.1% and praziquantel 8.6% (Profender®, Bayer), which is licensed for treatment of the more common cat lungworm Aelurostrongylus abstrusus, for the treatment of natural troglostrongylosis. Methods Sixteen cats enrolled in the study were 1:1 allocated to two groups, i.e. Group T, treated with Profender® spot-on on days 0 and 14 (± 2) at the recommended clinical dose, and Group C which remained untreated. After study completion, the control cats received two rescue treatments with Profender® on days 28 (± 2) and 42 (± 2). The primary efficacy criterion was the absence of T. brevior L1 following treatment. Other efficacy parameters were the quantitative comparison of L1 presence before (baseline) and after treatment in both groups, and the comparison of clinical signs pre- and post-treatment. Results In terms of stopping larval shedding, Profender® showed an efficacy of 97% and 97.5% (arithmetic and geometric means, respectively) for group T, 97.1% and 98.5% for group C after one administration, and 100% for both groups after two doses. Overall, 12 cats showed clinical signs related to T. brevior. Specifically, 9 were clinically affected before treatment while clinical signs appeared after the first treatment in 3 cats. At the end of the study, all symptomatic cats fully recovered with the exception of 3 cats that showed clinical signs similar to those observed at the pre-treatment examination at the end of the study. Conclusions This study shows that Profender® is effective against T. brevior.

Extra-intestinal nematodes of companion animals are of growing concern in veterinary medicine for their pathogenic potential and the current expansion throughout Europe. The present study has evaluated the occurrence of major canine and feline extra-intestinal nematodes in regions of Italy having epidemiological relevance. Associations of various recorded parameters related to the examined animals have been statistically evaluated, along with a comparative analysis with the most recent epidemiological data. Overall, 1055 dogs and 1000 cats were tested. Among extra-intestinal nematodes Angiostrongylus vasorum was the most common in dogs followed by Capillaria aerophila and Dirofilaria spp.; Aelurostrongylus abstrusus was the most recorded parasite in cats, followed by C. aerophila and Troglostrongylus brevior. The statistical analysis revealed that outdoor access is associated with A. vasorum, A. abstrusus and T. brevior infections, that were also more prevalent in animals with cardio-respiratory signs. Moreover, cats aged less than 12 months had more chances to be infected by lungworms. The data herein presented confirm the occurrence and the possible risk of expansion of different extra-intestinal parasitoses of dogs and cats in Italy, underlining the importance of a constant epidemiologic vigilance and of appropriate control methods.

International audience; Dirofilaria repens is a zoonotic, mosquito-borne filaria infecting carnivores, particularly dogs. It is expanding its range in Europe but epidemiological information is sparse for other Eurasian regions. In Hong Kong and India, the closely related species Candidatus Dirofilaria hongkongensis was proposed. Previous analysis of 2.5 kb partial mitochondrial genome sequences containing the particularly variable non-coding control region revealed low diversity in European D. repens while Asian nematodes showed high diversity. Sequences derived from feline blood samples from Narathiwat (Thailand) led to the proposal of a third potential species, Dirofilaria sp. "Thailand II". To avoid bias from rapidly evolving non-coding regions, this study aimed to compare Dirofilaria sp. "Thailand II" with D. repens and C. D. hongkongensis based on complete mitochondrial genomes. Using PCRs and Sanger sequencing, three complete mitochondrial genomes (13,651 bp) were assembled from DNA obtained from different feline blood samples. Mitochondrial genome organization was identical to other onchocercids with eleven protein-coding, two rRNA and 22 tRNA genes and no atp-8 gene. All genes were on the same strand showing an extremely high thymidine content (56.7%). Maximum-likelihood phylogenetic analysis using protein and rRNA sequences confirmed closer relationship of Dirofilaria sp. "Thailand II" to C. D. hongkongensis than to D. repens. All distances between these three putative species were considerably larger than the distance between the valid sibling species Onchocerca volvulus and Onchocerca ochengi. Sequencing of a 2.5 kb fragment containing the control region from microfilarial DNA from additional feline blood samples from Narathiwat 3-4 years later revealed that these also fell into the C. D. hongkongensis clade but were remarkably different from C. D. hongkongensis and Dirofilaria sp. "Thailand II". Since D. repens-like filaria are absent from dogs in Narathiwat, further field studies are required to confirm if these genotypes represent locally circulating cat-specific Dirofilaria genotypes or species.

Thelazia callipaeda (Spirurida, Thelaziidae) has been documented as agent of ocular infection in domestic animals (dogs and cats), wildlife (e.g., foxes, hares, rabbits), and humans. In the last two decades, this parasitosis has been increasingly reported in several European countries. Both adult and larval stages of the eyeworm are responsible for symptoms ranging from mild (e.g., lacrimation, ocular discharge, epiphora) to severe (e.g., conjunctivitis, keratitis, and corneal opacity or ulcers). The present study evaluated the clinical efficacy and safety of imidacloprid 10% and moxidectin 2.5% spot on (Advocate®, Bayer Animal Health) in comparison to milbemycin oxime/praziquantel tablets (Milbemax®, Novartis-Animal Health), as positive control, in the treatment of canine thelaziosis in naturally infected dogs and, a third group was used as an untreated control. Forty-seven dogs (27 females and 20 males) harbouring at least one live adult worm of T. callipaeda in one eye were enrolled from an endemic area of southern Italy. Each dog was then weighed and assigned in accordance with a random treatment allocation plan to one of the treatment groups (G1: imidacloprid 10% and moxidectin 2.5% spot on, G2: Untreated control and G3: milbemycin oxime/praziquantel tablets). On Day (D) 7, 14, 28 and 35 dogs were physically examined and the infection level was assessed by examination of both eyes, including conjunctival pouch and third eyelid for live adult T. callipaeda count and clinical scores. Dogs in G1 were treated on D0 and D28, whereas those in G3 on D0 and D7. Efficacy in G1 was 100% at each day post treatment (p

Hepatozoon canis is a blood parasite of the suborder Adeleorina infecting wild and domestic canids. Transmission occurs by oral uptake of Rhipicephalus sanguineus sensu lato vector ticks infected with H. canis, but vertical transmission is also assumed to be possible. In German foxes, a high prevalence of H. canis has previously been reported despite the fact that R. sanguineus s.l. is not endemic. In the absence of knowledge about local transmission pathways, foxes should be considered to be possible reservoirs of H. canis and contribute to infection of domestic dogs. The present study aimed to determine how often foxes and dogs are infected in Brandenburg (Germany) and if identical or different H. canis 18S rRNA haplotypes are found in these host species. Hepatozoon spp. were detected by PCR in 46/1050 (4.4 %) of dog blood and 176/201 (77.6 %) of fox spleen samples from Brandenburg. Sequencing of 19 dog and 56 fox samples identified all as H. canis. For nine positive dogs, owners stated that they had never left Germany suggesting that autochthonous transmission occurs not only in foxes but also in dogs. Sequences for seven of these possible autochthonous cases were obtained and six were identical to the predominant haplotype found in the foxes. Haplotype network analysis confirmed that many dogs, including some without travel history, carried the same or very similar 18S rRNA haplotypes as the foxes suggesting that both hosts participate in the same epidemiological cycle.

Angiostrongylus vasorum is a nematode of the superfamily Metastongyloidea and was first described in the south of France in the nineteenth century where its biology was extensively studied in the 1960ies, leading to the common use of the byname "the French heartworm". The life cycle of the parasite is indirect including slugs and snails as intermediate hosts, while the adults live in the right heart and pulmonary arteries. Since then, many studies have been conducted in different European countries. However, there is scant knowledge regarding the prevalence and distribution of this parasite in Greece. Therefore, the objective of the present study was to investigate the current seroprevalence of A. vasorum in dogs in Greece. Overall 1000 apparently healthy and randomly selected dogs, presented in veterinary clinics, were involved at national level (n = 66 locations). Individual serum samples were obtained and tested using two in-house ELISAs by combining the detection of specific antibodies against the parasite and of circulating antigens of A. vasorum. Overall, 3.0% (n = 30, CI 95% 2.1 to 4.3%) of the sampled dogs were seropositive to at least one of the two ELISAs: 0.4% (n = 4, CI 95% 0.2 to 1.0%) were positive by both ELISAs, while 1.0% (n = 10, CI 95% 0.5 to 1.8%) were seropositive only for specific antibodies and 1.6% (n = 16, CI 95% 1.0 to 2.6%) were antigen-positive only. This is the first multicentric seroepidemiological study about A. vasorum in dogs from Greece. We showed that this parasite circulates in the local canine population and further studies should be carried out to elucidate its significance in the country.

Metastrongyloid parasites represent sparsely studied parasites of dogs and cats in Germany. Recent European surveys indicate that these parasites are spreading in Europe. Actual data on prevalence of Angiostrongylus vasorwn in dogs and foxes reveal several endemic foci in Germany. However, actual data on the prevalence of A. vasorum and other metastrongyloid lungworm larvae in a wide range of slug and snail intermediate hosts, such as Anion lusitanicus, are missing for Germany. To fill this gap, we conducted an epidemiological survey on native German slugs in selected regions of Hesse and Rhineland-Palatinate. The focus was on slugs, because in study areas slugs appear to be more abundant than snails. Slugs were collected throughout different seasons of the year in areas that were previously proven to be hyperendemic for A. vasorum fox infections. Overall, a total of 2701 slugs were collected and examined for lungworm larvae via artificial digestion. The number of A. vasorum larvae per slug varied considerably (1-546 larvae per specimen). Some hotspot areas with high A. vasorum prevalence in slugs (up to 19.4%) were identified. The overall A. vasorum prevalence varied with season with largest number of slugs infected in summer (9.1%) and lowest number in winter (0.8%). The current study revealed a total A. vasorum prevalence of 4.7% in slugs based on microscopic analyses. Confirmation of lungworm species was made by specific duplex-real-time PCRs. Hence, these data demonstrate that final hosts are at a permanent risk for A. vasorum infections during all seasons when living in investigated areas. Besides A. vasorum, other lungworm larvae were also detected, such as Crenosoma vulpis (the fox lungworm, 2.3%) and Aelurostrongylus abstrusus (feline lungworm, 0.2%).

The efficacy of anthelmintic treatment at 1, 3, and 6 month intervals was evaluated in a prospective controlled field study with naturally exposed Lithuanian village dogs by monthly coproscopy during 1 year. A placebo-treated control group (C) (n = 202) and groups treated with two broad-spectrum anthelmintics, febantel/pyrantel-embonate/praziquantel (Drontal® Plus, Bayer) (D1, D3, D6; n = 113–117) and emodepside/praziquantel (Profender®, Bayer) (P1, P3, P6; n = 114–119), were included. At the beginning of the study, eggs of Toxocara canis (4.02%) and T. cati (0.44%) identified morphometrically and/or molecularly and eggs of taeniid- (0.78%) and Capillaria-like eggs (5.03%) were present in the feces without significant differences in prevalence between groups. Significant decreases in excretion of T. canis eggs was found 1 month after the treatment with Drontal® Plus in February (D1) and with Profender® in October (P1), November (P1), December (P3), February (P1), and March (P1, P3), as compared to controls in the same months. The incidence of egg excretion per dog at least once a year was significantly lower in group P1 for T. canis (4.24%; p

In recent years Angiostrongylus vasorum has become another important heart parasite of dogs besides Dirofilaria immitis, with intense spread into new areas of Europe. The first two cases of canine angiostrongylosis in Slovakia were observed in 2013, demonstrating that this life-threatening parasitic disease of dogs has expanded into this territory too. One year after the first A. vasorum findings, a serological survey was conducted to assess the current distribution of this parasitic infection in dogs from Slovakia. Serum samples from 225 dogs were collected from 29 veterinary practices situated in 22 districts of Slovakia and tested by ELISA for the presence of circulating A. vasorum antigens and additionally for the detection of specific antibodies against A. vasorum. Fourteen samples (6.22 %) were seropositive in at least one ELISA. Of these, 7 dogs (3.11 %) were only antibody-positive and 4 dogs (1.78 %) were positive only for circulating A. vasorum antigen. Three animals out of 225 examined (1.33 %) were positive in both ELISAs

Canine vector-borne diseases (CVBDs) have increasingly become a focus of interest in recent years. Some of the CVBDs are zoonotic and may therefore also represent a risk for the human population. Different factors are in discussion to explain the expansion of vectors and pathogens into formerly unaffected areas. Knowledge of the prevalence and distribution of CVBDs in Bulgaria is scant overall and most data rely on single case descriptions. The aim of the present study was to determine the seroprevalence of important CVBDs in 167 dogs from central-southern Bulgaria (Stara Zagora), with special emphasis on hitherto uninvestigated babesiosis and angiostrongylosis, on poorly investigated Lyme borreliosis and canine granulocytic anaplasmosis, and on the potentially zoonotic dirofilariosis and leishmaniosis. Relatively high prevalence rates were documented for anti-Babesia canis antibodies, Dirofilaria immitis antigen (16.2 %; 27/167 each), anti-Ehrlichia canis (21 %; 35/167) and anti-Anaplasma phagocytophilum antibodies (30.5 - 46.1 %; 51 - 77/167), while Borrelia burgdorferi seroprevalence was low (2.4 %; 4/167). All samples were negative for Leishmania infantum antibodies and Angiostrongylus vasorum antigen and antibodies. In total, 64.7 % (108/167) of the samples indicated infection or exposure to at least one agent and a high proportion of dual infections (39.8 %; 43/108) was demonstrated. Multiple infections with up to four different organisms were also detected. Our data underline the importance of CVBDs and especially of co-infections which could influence the clinical outcome in dogs.

This study aimed to evaluate the efficacy of an emodepside 2.1 % (w/v)/praziquantel 8.6 % (w/v) topical solution (Profender® spot-on for cats) in the prevention of lactogenic Toxocara cati infections. A controlled test was performed with two groups of 8 cats with confirmed pregnancy. All cats were infected with daily doses of 2000 T. cati eggs for 10 consecutive days starting 50 days post conception to produce an acute infection. Treatment was performed 60 days post conception. Queens in the treatment group received the emodepside/praziquantel solution at the minimum therapeutic dose (3 mg/kg emodepside and 12 mg/kg praziquantel), while the control group was treated with a placebo spot-on. Efficacy was evaluated 56 days post partum by necropsy of one randomly selected kitten of each litter and comparison of the worm burdens between the study groups. Additionally the necropsy results were supported by quantification of worms expelled with the faeces after deworming of the remaining kittens and all queens. The treatment in late pregnancy resulted in an efficacy of 98.7 % (p < 0.0001). All necropsied control kittens were infected (geometric mean 30.6). Seven of 8 kittens from treated mothers were free of T. cati (geometric mean 0.4). Worm counts after deworming reflected the results obtained at necropsy. No side effects of the treatment were observed. It is concluded that treatment with an emodepside/praziquantel spot-on solution during late pregnancy effectively prevents lactogenic transmission of T. cati to the offspring. The study design facilitated the generation of reliable data, while at the same time a minimum number of animals was sacrificed.

SUMMARYDogs infected with Angiostrongylus vasorum, a potentially lethal parasite parasitizing the heart and pulmonary arteries, may present severe respiratory, haematological and neurological signs. In this first large-scale seroepidemiological survey, 4003 sera originating from Germany and 4030 from the UK were tested by an ELISA for the detection of circulating antigen of A. vasorum, and by a separate ELISA detecting specific antibodies. In Germany, where mainly western federal states were sampled, 0·3% (n = 13, CI: 0·2–0·6%) of dogs were positive in both ELISAs, whereas in total 0·5% (n = 20, CI: 0·3–0·8%) were antigen-positive and 2·25% (n = 90, CI: 1·8–2·8%) were positive for specific antibodies. Regions with antigen- and antibody-positive animals were overlapping. In the UK, where mainly the south of the country was sampled, 0·97% (n = 39, CI: 0·7–1·3%) of dogs were antigen- and antibody positive. In total, 1·32% (n = 53, CI: 1·0–1·7%) were antigen-positive, and 3·2% (n = 129, CI: 2·7–3·8%) were positive for specific antibodies, again in overlapping regions. These results confirm the occurrence of A. vasorum in a random dog population originating from large parts of the countries investigated. The use of the tests alone or in combination was considered as a function of their sensitivities and specificities, in order to guide efficient clinical and epidemiological application.