The 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. Faecal 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. Totals 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. High 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.

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.