BackgroundThe objectives of this study were to evaluate the prevalence and infection intensity of intestinal parasites in different dog and cat populations in Crete, Greece, estimate the zoonotic risk and identify risk factors.MethodsFaecal samples from shelter, household and shepherd dogs and shelter and household cats were analyzed using sedimentation/flotation techniques. Giardia and Cryptosporidium were detected by a quantitative direct immunofluorescence assay (IFA). PCR and sequencing was performed to evaluate the zoonotic potential of Giardia and Cryptosporidium positive samples.ResultsTotals of 879 dog and 264 cat faecal samples were examined. In dogs, the overall prevalence was 25.2% (CI: 22.4–28.1) for Giardia spp.; 9.2% (CI: 7.3–11.1) for Ancylostoma/Uncinaria spp.; 7.6% (CI: 5.9–9.4) for Toxocara spp.; 5.9% (CI: 4.4–7.5) for Cryptosporidium spp.; 4.6% (CI: 3.2–5.9) for Cystoisospora spp.; 2.7% (CI: 1.7–3.8) for Toxascaris leonina; 1.7% (CI: 0.9–2.6) for Capillaria spp.; 0.8% (CI: 0.2–1.4) for taeniid eggs; 0.2% (CI: 0–0.5) for Dipylidium caninum; and 0.1% (CI: 0–0.3) for Strongyloides stercoralis. In cats, the prevalence was 20.5% (CI: 15.6–25.3) for Giardia spp.; 9.5% (CI: 5.9–13.0) for Cystoisospora spp.; 8.3% (CI: 5.0–11.7) for Toxocara spp.; 7.6% (CI: 4.4–10.8) for Ancylostoma/Uncinaria spp.; 6.8% (CI: 3.8–9.9) for Cryptosporidium spp.; 4.2% (CI: 1.8–6.6) for Capillaria spp.; 0.8% (CI: 0–1.8) for taeniid eggs; and 0.4% (CI: 0–1.1) for Hammondia/Toxoplasma. Concerning the risk factors evaluated, there was a negative association between age and Giardia infection and between age and T. leonina infection intensity for dogs. Sequencing results revealed the presence of mainly animal-specific G. duodenalis assemblages C and D in dogs and assemblages F, C and BIV-like in cats, with only a limited number of (co-)infections with assemblage A. As for Cryptosporidium, the dog-specific C. canis and the pig-specific C. scrofarum were detected in dogs and the cat-specific C. felis was detected in cats.ConclusionsHigh levels of parasitism in both dogs and cats were recorded. Giardia was the most prevalent parasite in all dog and cat populations except for shepherd dogs. Genotyping results suggest a limited zoonotic risk of Giardia and Cryptosporidium infections from dogs and cats in Crete. Taeniid eggs were more prevalent in shepherd dogs suggesting access to carcasses and posing a threat for cystic echinococcosis transmission. Infection rates of Toxocara spp. in both dogs and cats show that companion animals could be a significant source of infection to humans.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-1989-8) contains supplementary material, which is available to authorized users.

Background: The aim of this study was to determine exposure to vector-borne pathogens (VBPs) in populations of dogs living on Greek islands in the Ionian and Aegean seas. Methods: In total, 1154 dogs with different lifestyles and of varying ages and breeds were randomly sampled and examined for the presence of clinical signs compatible with canine vector-borne diseases (CVBDs). Blood was collected from each individual animal. For the detection of antibodies against Leishmania spp., the WITNESS ® Leishmania test was performed, and positive samples were further examined with indirect enzymatic immunoassay (ELISA). Antibodies to Borrelia burgdorferi, Ehrlichia canis or E. ewingii, as well as Anaplasma phagocytophilum or A. platys were investigated using the Snap ® 4Dx ® Plus test. Positive Ehrlichia spp. and Anaplasma spp. samples were further examined using an indirect ELISA for further identification of the species. Results: In total, 25.6% of dogs were exposed to at least one of the pathogens investigated, with seroprevalences varying regionally. Of these seropositive dogs, 27.4% displayed clinical signs suggestive of CVBDs, such as cutaneous lesions, enlarged lymph nodes, pale mucous membranes, onychogryphosis and weight loss. The overall seroprevalence detected using the rapid tests was 15.3% for Leishmania spp., whereas 2.3% of the examined dogs were found to be positive for Anaplasma spp. and 7.5% for Ehrlichia spp. while B. burgdorferi was not detected. Twenty-four samples positive to A. phagocytophilum by ELISA were analysed by PCR for the presence of Anaplasma DNA. PCR and sequencing results showed the presence of A. platys DNA in 4 samples and E. canis DNA in 4 samples. The remaining samples (66.7%) were negative. Conclusions: In the present study, exposure of dogs to VBPs was shown in the geographical areas investigated. Results confirm that on Greek islands VBPs represent a constant health risk for both native and visiting dogs, suggesting the presence of distinct "hot-spots" of VBP infections on different islands. In order to reduce the risk of transmission and the spread to non-endemic regions, the protection of dogs through use of repellents and vaccines, together with owner education, seem to be of paramount importance.

Phlebotomine sandflies are vectors of the humans’ and mammals’ parasite Leishmania spp. Although the role of gut microbiome in the biological cycle of insects is acknowledged, we still know little about the factors modulating the composition of the gut microbiota of sandflies. We tested whether host species impose a strong structural effect on the gut microbiota of Phlebotomus spp. Sandflies were collected from the island of Leros, Greece, and classified to P. papatasi, P. neglectus, P. tobbi, and P. similis, all being negative to Leishmania spp. The prokaryotic gut microbiota was determined via 16S rRNA gene amplicon sequencing. Phlebotomus species supported distinct microbial communities (p < 0.001). P. papatasi microbiota was the most distinct over-dominated by three Spiroplasma, Wolbachia and Paenibacillus operational taxonomic units (OTUs), while another Wolbachia OTU prevailed in P. neglectus. Conversely, the microbiota of P. tobbi and P. similis was composed of several less dominant OTUs. Archaea showed low presence with the dominant OTUs belonging to methanogenic Euryarcheota, ammonia-oxidizing Thaumarcheota, and Nanoarchaeota. We provide first insights into the composition of the bacterial and archaeal community of Phlebotomus sandflies and showed that, in the absence of Leishmania, host genotype is the major modulator of Phlebotomus sandfly gut microbiota.

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Esafoxolaner, a purified enantiomer of afoxolaner with insecticidal and acaricidal properties, is combined with eprinomectin and praziquantel, nematodicidal and cestodicidal compounds, in NexGard® Combo, a novel topical endectoparasiticide formulation for cats. The efficacy of this formulation was assessed against Otodectes cynotis in two laboratory studies conducted in South Africa and in the USA with local isolates, and in one field trial conducted in Europe. In each study, cats were randomly allocated to a placebo-treated control group and a novel formulation-treated group. In the laboratory studies, cats were treated at the minimum recommended dose; in the field trial, cats were treated at label dose. All included cats were diagnosed positive for O. cynotis prior to treatment by otoscopy. The main variable of efficacy was a comparison of the number of live O. cynotis collected in both ear canals of all cats in the treated and control groups, one month after treatment. Efficacy of the novel topical formulation exceeded 97% in the three studies. These studies demonstrated the high effectiveness of NexGard® Combo in cats for the treatment of O. cynotis infestations. No health abnormalities were attributed to the treatment in any of the studies.