The detection of atypical Kinetoplastida in vertebrate hosts and vectors might suggest unexpected host-parasite contacts. Aside to major vectors of Leishmania (Leishmania) infantum in Italy (e.g. Phlebotomus perniciosus and Phlebotomus perfiliewi), the sand fly fauna also includes Sergentomyia minuta, herpetophilic and proven vector of Leishmania (Sauroleishmania) tarentolae, in which records of blood meal on mammals and detection of L. infantum DNA are increasing. This study was conducted in Central Italy aiming to molecularly detect potential atypical Leishmania host-vector contacts. Detection of Leishmania spp. DNA was performed by polymerase chain reaction (SSU rRNA, ITS1 targets) on field-collected sand fly females (N = 344), blood samples from humans (N = 185) and dogs (N = 125). Blood meal identification was also performed on engorged sand flies. Leishmania spp. DNA was found in 13.1% sand flies, 3.7% humans and 14.4% dogs. Sequence analysis identified L. infantum in S. minuta (4.4%), P. perniciosus (9.1%), humans (2.2%) and dogs (14.4%). Leishmania tarentolae was detected in S. minuta (12.6%), P. perfiliewi (6.6%) and human (1.6%) samples. Of 28 S. minuta examined for blood meal, 3.6 and 21.4% scored positive for human and lizard DNA, respectively. These results indicate the importance of one-health approach to explore new potential routes of transmission of leishmaniasis involving S. minuta.

Dirofilaria immitis and Dirofilaria repens are mosquito‐borne filarioids of zoonotic concern, which primarily cause canine heartworm disease (HWD) and subcutaneous dirofilariosis, respectively. The geographical distribution of these nematodes is constantly changing, due to many factors such as the colonization by new invasive mosquito species, the increased movement of animals and the absence of chemoprophylactic strategies in non‐endemic regions. Although HWD has always been considered endemic in northern Italy, an increase in the prevalence of this disease has been recorded in the last decades in central and southern regions. We describe the southernmost hyperendemic European focus of heartworm disease in the Pelagie archipelagos. From June to November 2020, 157 dogs and 46 cats were sampled in Linosa and Lampedusa islands for the detection of Dirofilaria immitis and Dirofilaria repens by modified Knott's, rapid antigen and molecular tests. Mosquitoes were collected using CDC light and BG sentinel‐2 traps and aspirators, and tested for host blood meal and Dirofilaria spp. DNA. Out of 56 dogs in Linosa and 101 in Lampedusa, 33 (i.e. 58.9%) and 8 (i.e. 7.9%) were positive to D. immitis and D. repens, respectively. Three cats scored positive to D. immitis (i.e. 17.6%) in Linosa. Six mosquito species were identified, and the abdomen of a non‐engorged Aedes albopictus was positive for D. immitis and human DNA. The results suggest that D. immitis infection could spread to new previously non‐endemic territories in southern Europe, representing a real threat to animal and human health.

Reptile vector-borne diseases (RVBDs) of zoonotic concern are caused by bacteria, protozoa and viruses transmitted by arthropod vectors, which belong to the subclass Acarina (mites and ticks) and the order Diptera (mosquitoes, sand flies and tsetse flies). The phyletic age of reptiles since their origin in the late Carboniferous, has favored vectors and pathogens to co-evolve through millions of years, bridging to the present host-vector-pathogen interactions. The origin of vector-borne diseases is dated to the early cretaceous with Trypanosomatidae species in extinct sand flies, ancestral of modern protozoan hemoparasites of zoonotic concern (e.g., Leishmania and Trypanosoma ) associated to reptiles. Bacterial RVBDs are represented by microorganisms also affecting mammals of the genera Aeromonas, Anaplasma, Borrelia , Coxiella , Ehrlichia and Rickettsia , most of them having reptilian clades. Finally, reptiles may play an important role as reservoirs of arborivuses, given the low host specificity of anthropophilic mosquitoes and sand flies. In this review, vector-borne pathogens of zoonotic concern from reptiles are discussed, as well as the interactions between reptiles, arthropod vectors and the zoonotic pathogens they may transmit.

Leishmaniasis (or the leishmaniases), classified as a neglected tropical parasitic disease, is found in parts of the tropics, subtropics and southern Europe. Leishmania parasites are transmitted by the bite of phlebotomine sand flies and million cases of human infection occur annually. Leishmania tarentolae has been historically considered a non‐pathogenic protozoan of reptiles, which has been studied mainly for its potential biotechnological applications. However, some strains of L. tarentolae appear to be transiently infective to mammals. In areas where leishmaniasis is endemic, recent molecular diagnostics and serological positivity to L. tarentolae in humans and dogs have spurred interest in the interactions between these mammalian hosts, reptiles and Leishmania infantum, the main aetiologic agent of human and canine leishmaniasis. In this review, we discuss the systematics and biology of L. tarentolae in the insect vectors and the vertebrate hosts and address questions about evolution of reptilian leishmaniae. Furthermore, we discuss the possible usefulness of L. tarentolae for new vaccination strategies.

Leishmania tarentolae is a nonpathogenic trypanosomatid species isolated from geckoes in the Mediterranean basin. In Italy, L. tarentolae and Leishmania infantum occur in sympatry in areas where canine leishmaniosis (CanL) is endemic. Here, we investigated the prevalence and distribution of L. tarentolae in reptilian hosts and the presence of L. infantum in the same animals in southern Italy. From April 2020 to April 2021, lizards and snakes were captured in urban, peri-urban sites, natural parks, and dog shelters. Blood was individually sampled and assessed for the presence of Leishmania spp. by cytology and/or molecular biology. Captured lizards were euthanized and their tissues/organs were tested by duplex real-time-PCR for the detection of L. infantum and L. tarentolae and the positive ones were amplified by conventional PCR to confirm the species identification by sequencing. Samples were also screened by realtime PCR for L. infantum kDNA minicircle. Of the 259 collected Squamata reptiles, 34 (13.1%) specimens scored positive for Leishmania spp., with a prevalence of 10% for L. tarentolae (n = 26) and of 3.1% for L. infantum (n = 8). Positive lizards were mostly Podarcis siculus (n = 26) from dog shelters, with six specimens positive for L. infantum. In addition, L. tarentolae was detected in one Podarcis filfolensis lizard, and in five T. mauritanica geckos (with one gecko and P. filfolensis lizard showing amastigote-like forms in thrombocytes), whereas L. infantum was detected in one T. mauritanica, and one Hemidactylus turcicus. Data herein reported demostrate that saurian reptiles living in CanL endemic areas of the Mediterranean basin may be exposed to both L. tarentolae and L. infantum indicating a sympatric circulation of these species in different vertebrate hosts. Moreover, this study represents the first molecular evidence of L. infantum infection in European reptiles.

The trypanosomatid protist Leishmania tarentolae is a saurian-associated parasite vectored by the Sergentomyia minuta sand fly. This study aimed to confirm the circulation of L. infantum and L. tarentolae in sand flies, reptiles and dogs and to isolate new strains of these protists. Reptilian and sheltered dog blood samples were collected, and sand flies were captured. Samples were tested for Leishmania spp. using duplex real-time PCR (dqPCR) and real-time PCR (qPCR); the origin of blood meal was identified in engorged sand flies by conventional PCR. The reptilian blood and intestinal content of sand fly females were cultured. Dog sera were tested by IFAT using both Leishmania species. Four Tarentola mauritanica geckoes were molecularly positive for L. infantum or L. tarentolae, with no co-infections; moreover, amastigote-like forms of L. infantum were observed in the bone marrow. 24/294 sand flies scored positive for Leishmania spp. by dqPCR, 21 S. minuta and two Phlebotomus perniciosus were positive for L. tarentolae, while only a single Ph. perniciosus was positive for L. infantum. Blood meal analysis confirmed reptile and dog in S. minuta, dog and human in Ph. perniciosus and dog in Phlebotomus neglectus. Two axenic strains of L. tarentolae were obtained. Twelve of 19 dogs scored positive for L. infantum and L. tarentolae by IFAT and three of them also for L. infantum by dqPCR, and six by qPCR. These data confirm the sympatric circulation of L. infantum and L. tarentolae in geckoes, sand flies, and dogs, and suggest that geckoes may be infected with L. infantum.

Background Vector-borne diseases (VBDs) represent an emerging global threat to public health due to the geographical expansion of arthropod vectors. The study aims to assess the seroprevalence of selected vector-borne pathogens (VBPs) in different groups of outdoor workers and the occupational risk factors for exposure to arthropod bites. Methods A cross-sectional study was conducted on 170 workers recruited in two different regions of southern Italy, including farmers, forestry workers, veterinarians, geologists/agronomists and administrative employees, and tested for IgG antibodies against Bartonella henselae, Borrelia spp. Coxiella burnetii and Rickettsia conorii, using a chemiluminescent immunoassay (CLIA). The relationship among job characteristics, tick exposure and the prevalence of seropositive subjects for each pathogen was investigated by applying categorical principal component analysis (CATPCA). Results A high seroprevalence for C. burnetii (30.0%) and R. conorii (15.3%) was reported, mainly in farmers (67.7% and 54.8%, respectively) and forestry workers (29.0% and 16.1%, respectively), while a low prevalence was observed for B. henselae and Borrelia spp. (8.8% and 4.1%, respectively). The regression equation by CATPCA was significant for C. burnetii and R. conorii (P < 0.001), showing a positive association with job, tick bite exposure, working area and contact with animals. Conclusions These findings highlight the need of activating an appropriate occupational health response for minimizing the risk of arthropod vector exposure in workplaces, considering specific preventive measures in particular in high-risk job categories. Graphical Abstract

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.