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Additional file 1: Figure S1. Overview of the moments of sampling within the average seasonal variation in precipitation and temperature. Table S1. Different multiplex assays for the detection of tick- and flea-borne pathogens. Table S2. Accession numbers for tick and flea species identified. Table S3. Distribution of PCR signals allocated to an ectoparasite taxon (identification at genus level and more precise) in the infested cats of urban and rural areas. Table S4. Distribution of co-infested cats within the subpopulation of tick-infested cats. Table S5. Co-infestations by different flea species (identification at genus level and lower). Table S6. Co-infections in cat blood. Table S7. Co-infections in cat ticks. Table S8. Co-infections in cat fleas.

Additional file 2. Capture form.

Additional file 2. Capture form.

Additional file 1: Table S1. Most abundant bacteria not suspected to be vector-borne pathogens. Bacterial species, genera and families detected via our NGS methodology that are not suspected pathogens across all samples as a percentage of total reads that passed filtering. Mycoplasma spp., E. canis and A. platys were all more abundant than these commensal or contaminant bacterial groups.

Additional file 1: Fig. S1. Overview of sampling times and average seasonal variation in precipitation and temperature. Table S1. Distribution of PCR signals allocated to an ectoparasite taxon (identification at genus level and more precise) in the infested dogs of urban and rural areas. Table S2. Distribution of co-infested dogs within the subpopulation of tick-infested dogs. Table S3. Co-infestations by different flea species (identification at genus level and lower). Table S4. Co-infections in dog blood. Table S5. Co-infections in dog ticks. Table S6. Co-infections in dog fleas. Table S7. Correlations with sero-prevalences. Table S8. Correlations with flea-borne pathogens.

Additional file 1: Fig. S1. Overview of sampling times and average seasonal variation in precipitation and temperature. Table S1. Distribution of PCR signals allocated to an ectoparasite taxon (identification at genus level and more precise) in the infested dogs of urban and rural areas. Table S2. Distribution of co-infested dogs within the subpopulation of tick-infested dogs. Table S3. Co-infestations by different flea species (identification at genus level and lower). Table S4. Co-infections in dog blood. Table S5. Co-infections in dog ticks. Table S6. Co-infections in dog fleas. Table S7. Correlations with sero-prevalences. Table S8. Correlations with flea-borne pathogens.

Background: Arthropod-borne pathogens and their vectors are present throughout Africa. They have been well studied in livestock of sub-Saharan Africa, but poorly studied in companion animals. Given their socioeconomic importance, the African Small Companion Animal Network (AFSCAN), as part of the WSAVA Foundation, initiated a standardized multi-country surveillance study. Methods: In six countries (Ghana, Kenya, Nigeria,Tanzania, Uganda, and Namibia) in both rural and urban settings, 160 infested cats were sampled to assess their ectoparasite community (ticks and fleas), as well as the micro-parasite prevalence within those ectoparasites (60 and 118 pools of ticks and fleas, respectively) and blood (276 cats, including 116 non-infested). Results: Almost two thirds of all infested cats originated from Tanzania and Kenya. Despite the large macro-geographical variation, no consistent difference was found in ectoparasite diversity and numbers between East and West Africa. Far more flea-infested than tick-infested cats were found. The most dominant ectoparasite was Ctenocephalides felis. Among the ticks, the exophilic Haemaphysalis spp. were the commonest, including species that are not typically linked with companion animals (Haemaphysalis spinulosa and Haemaphysalis elliptica). The most prevalent pathogens found in the blood and fleas were Bartonella henselae and Mycoplasma haemofelis. In the ticks, the dog-associated Hepatozoon canis was most commonly found. A high degree of co-parasitism was found in all countries and habitats. Conclusions: Our continent-wide standardized field study highlights the cat's potential to serve as a reservoir of pathogens that can be transmitted to humans or livestock, especially when cats are expected to become more commonly kept in African villages and towns. DH is funded by the Marie Sklodowska-Curie Actions (EU-Horizon 2020, Individual Global Fellowship, Project No. 799609). The study was funded by Elanco Animal Health, within the umbrella of the African Small Companion Animal Network (AFSCAN) programme of the World Small Animal Veterinary Association (WSAVA) and supported by IDEXX Laboratories and Clinvet International (Pty) Ltd. The funders had no role in the study design, data collection, interpretation and analysis, decision to publish, or preparation of the manuscript. We thank Dr. Sherry Johnson (Ghana), Dr. Samuel Maina Githigia (Kenya), Dr. Foluke Adedayo Akande (Nigeria), Dr. Jahashi Saidi Nzalawahe (Tanzania), Dr. Tayebwa Dickson (Uganda), and Dr. Ortwin Aschenborn (Namibia) for their technical assistance. This paper was sponsored by Elanco Animal Health in the framework of the 16th CVBD® World Forum Symposium.

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.