Explore the vast range of titles available.

Background Borrelia burgdorferi ( sensu lato ) is a complex containing pathogenic bacteria of which some species, such as Borrelia lusitaniae , use birds, small mammals and reptiles as reservoirs. In Italy, the bacteria have been detected in reptilian and avian reservoirs in the northern and central regions. Results Here, 211 reptiles from three orders [Squamata (Sauria with seven species in five families and Ophidia with 11 species in three families), Crocodylia (one family and two species), and Testudines (two families and two species)] were examined for ectoparasites and molecular detection of B. burgdorferi ( s.l. ) in three different sites of southern Italy, an area for which no information was previously available on the occurrence of borreliosis in animals and humans. Borrelia lusitaniae was molecularly detected in larvae and nymphs (11.6%) of Ixodes ricinus infesting lizards (i.e. Podarcis muralis , Podarcis siculus and Lacerta bilineata ) and in 12.3% blood samples of P. siculus . Finally, B. lusitaniae and Borrelia garinii were detected in 5.1% (32/630) of questing I. ricinus . Conclusions These results show the circulation of B. lusitaniae in southern Italy and suggest that P. siculus could play a role as a reservoir, representing a potential medical threat to humans living in or visiting these localities.

One of the most common behaviors of cats that have an indoor/outdoor lifestyle is to bring hunted “gifts” to their owners, represented by small mammals, reptiles and birds. Access to the outdoors by dogs and cats may represent a problematic issue, since they may be at risk of diseases, traffic accidents and ingestion of toxins. Yet, the impact of this population of roaming dogs and cats predating wildlife is another concerning issue that receives less attention. Despite these risks, most owners still prefer to give outdoor access to their pets to allow them to express their “natural instincts,” such as hunting. Therefore, with a growing population of > 470 million dogs and 373 million cats worldwide, predation not only represents a threat to wildlife, but also a door of transmission for parasitic diseases, some of them of zoonotic concern. In this review, the role played by dogs, and especially cats, in the perpetuation of the biological life cycle of zoonotic parasites through the predation of rodents, reptiles and birds is discussed. Feral and domestics dogs and cats have contributed to the population collapse or extinction of > 63 species of reptiles, mammals and birds. Although the ecological impact of predation on wild populations is well documented, the zoonotic risk of transmission of parasitic diseases has not received significant attention. The parasitic diseases associated to predation vary from protozoan agents, such as toxoplasmosis, to cestodes like sparganosis and even nematodes such as toxocariasis. Raising awareness about predation as a risk of zoonotic parasitic infections in dogs and cats will aid to create responsible ownership and proper actions for controlling feral and free-roaming cat and dog populations worldwide. Graphical Abstract

Background Ophionyssus natricis is the main species of mite that infests captive reptiles. High infestations may result in the host experiencing general discomfort and deleterious effects, even death. Moreover, O. natricis is an important vector of reptile vector-borne diseases and is considered to be the putative vector of the Reptarenavirus, the causal agent of the inclusion body disease. Despite the cosmopolitan distribution of O. natricis in captive reptiles, treatment options are limited. The aim of the present study was to assess the efficacy of afoxolaner (NexGard®; Boehringer Ingelheim, Ingelheim, Germany) in heavily infested, privately owned snakes, evaluate the prevalence of mites and drug availability in the plasma of treated snakes (pharmacokinetics) and perform a clinical examination of animals. Methods The study was conducted in two snake breeding facilities, where many snakes were infested with mites. Each animal was clinically examined and weighed, and mite infestations were assessed on the animals and in their enclosures (environment). Animals were treated with a dose of 2.5 mg afoxolaner per kilogram body weight (2.5 mg/kg) administered orally. All animals were examined pre-treatment (T0) and at various time points post-treatment (T1, 6 h; T2, 24 h; T3, 14 days; T4, 28 days). The collected mites were morphologically identified at the species level and the species identity also confirmed molecularly. Results Overall, 81 snakes from the two participating facilities (i.e. 70 from site 1 and 11 from site 2) were screened, and 31 (38.3%) snakes were found to have at least one mite. All mites were identified morphologically and molecularly as O. natricis. Lampropeltis was the genus of snakes with highest number of infested individuals. Mites were found to be alive on snakes at T1, but at T2 only dead mites were observed, and at T3 and T4 mites were no longer present on the animals or in their environment. No side effects were observed in the treated snakes. Conclusions A single oral administration of afoxolaner at 2.5 mg/kg was a safe treatment for snakes and 100% effective for the eradication of natural O. natricis infestation without the need to treat the environment of the snake. Graphical Abstract

Background Cats are now recognized as competent hosts for Leishmania infantum and a blood source for sand fly vectors. Although canine leishmaniosis (CanL) is endemic in Mediterranean Basin countries, large-scale epidemiological studies are lacking for feline leishmaniosis (FeL). This study aimed to assess the prevalence of L. infantum infections, associated risk factors, clinical signs, and clinicopathological abnormalities in domestic cat populations from six Mediterranean Basin countries. Methods From 2019 to 2022, blood and serum samples of cats ( n = 2067) living in Italy ( n = 300), Greece ( n = 297), Portugal ( n = 295), France ( n = 231), Israel ( n = 313), and Spain ( n = 631) were collected along with animal data (i.e., age, sex, breed, housing conditions, and geographical origin), clinical signs, and laboratory blood test parameters. Cats were grouped according to their age as kittens (up to 1 year), young (older than 1 and younger than 7 years), mature (between 7 and 10 years), and senior (older than 10 years). Serum samples were tested for L. infantum by immunofluorescence antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA), and blood samples of seropositive cats were tested for L. infantum kinetoplast deoxyribonucleic acid (kDNA). Viral infection by feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) was molecularly addressed in all cats enrolled. Statistical analysis was performed to evaluate the association between the risk of L. infantum infection and independent variables, and among co-infection of L. infantum with FIV and/or FeLV, clinical signs, and clinicopathological abnormalities. Results Overall, 17.3% (358/2067) of cats scored positive for L. infantum by serological tests. Specifically, 24.7% were from Portugal, 23.2% from Greece, 16.6% from Israel, 15% from Spain, 13.3% from France, and 12.6% from Italy. Leishmania infantum DNA was detected in 15 seropositive animals. Housing condition and FIV infection proved to be risk factors for FeL. Leishmania seropositivity was significantly associated with weight loss, lymphadenomegaly, gingivostomatitis, and oral ulcers, as well as with reduced albumin and albumin/globulin ratio, increased total globulins and total proteins, leukocytosis, and thrombocytosis. Conclusions This study provides, for the first time, a large-scale epidemiological survey on FeL and its clinical presentation, revealing that L . infantum circulates among domestic cats, especially shelter/free-roaming and FIV-infected animals, living in CanL endemic countries of the Mediterranean Basin. 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. Graphical Abstract

Visceral leishmaniasis (VL) caused by Leishmania infantum is endemic in the Mediterranean basin with most of the infected human patients remaining asymptomatic. Recently, the saurian-associated Leishmania tarentolae was detected in human blood donors and in sheltered dogs. The circulation of L . infantum and L . tarentolae was investigated in humans, dogs and cats living in the Pelagie islands (Sicily, Italy) by multiple serological and molecular testing. Human serum samples (n = 346) were tested to assess the exposure to L . infantum by immunofluorescence antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) and to L . tarentolae by IFAT. Meanwhile, sera from dogs (n = 149) and cats (n = 32) were tested for both Leishmania species by IFAT and all blood samples, including those of humans, by specific sets of real time-PCR for L . infantum and L . tarentolae . The agreement between serological tests performed for human samples, and between serological and molecular diagnostic techniques for both human and animal samples were also assessed. Overall, 41 human samples (11.8%, 95% CI: 8.9–15.7) were positive to L . infantum (5.2%, 95% CI: 3.3–8.1), L . tarentolae (5.2%, 95% CI: 3.3–8.1) and to both species (1.4%, 95% CI: 0.6–3.3) by serology and/or molecular tests. A good agreement among the serological tests was determined. Both Leishmania spp. were serologically and/or molecularly detected in 39.6% dogs and 43.7% cats. In addition to L . infantum , also L . tarentolae circulates in human and animal populations, raising relevant public health implications. Further studies should investigate the potential beneficial effects of L . tarentolae in the protection against L . infantum infection.

Leishmania is a neglected tropical parasite that may affect millions of people worldwide, causing infections that range from subclinical to life-threatening in many animal species, including humans. The four subgenera described (i.e., Leishmania, Mundinia, Sauroleishmania and Viannia) exhibit distinct biological and ecological characteristics. The genus likely originated during the Mesozoic era, with hypotheses suggesting a Palearctic, Neotropical, or Supercontinent origin. Understanding the evolutionary history of Leishmania parasites may clarify host specificity, geographic distribution, and vector associations across subgenera. In this study, we analyzed Leishmania evolutionary associations with vertebrate hosts as well as sand fly vectors using a combination of cophylogenetic and maximum parsimony approaches. A significant phylogenetic congruence was found between Leishmania and its vertebrate hosts using PACo (p = 0.0001, m2XY=0.5419) and ParaFit (p = 0.0034, ParaFit Global statistic = 0.012), with several host-switching and duplication events (p = 0.0298), particularly involving Leishmania-humans associations. A cospeciation event was also predicted at a higher taxonomic level, corresponding to the divergence of mammals from reptiles and the subsequent separation of the Leishmania, Viannia and Mundinia subgenera from Sauroleishmania. Similarly, significant cophylogenetic signals were observed between Leishmania and sand fly hosts using PACo (m2XY=1.3988, p = 0.0001) and ParaFit (ParaFit Global = 0.062, p = 0.0001) functions. Cospeciation events were again predicted at higher taxonomic levels in the divergence of New World and Old World sand flies, with Viannia species and Leishmania, Sauroleishmania and Mundinia species associated with the latter (p = 0.01). Our findings support the Supercontinent hypothesis and emphasize the relevance of the historical biogeography in Leishmania diversification. This comprehensive cophylogenetic analysis enhances our understanding of Leishmania origins and diversification, offering insights into host specificity, vector adaptation, and the long-term maintenance of its digenetic life cycle.

In canine leishmaniosis endemic areas, Leishmania infantum may occur in sympatry with the non-pathogenic Leishmania tarentolae , which is associated to reptiles. The potential infectivity of L . tarentolae for mammals raises questions about the interactions between the two Leishmania species, and the potential cross-immune protection in dogs. This study aimed to assess the outcome of experimental L . tarentolae infection in dogs, determining: i) the anti- L . tarentolae antibody production, ii) the duration of the immunity and cytokine expression, and iii) the possible pathogenic effect in the canine host. Twelve purpose-bred beagle dogs were randomly allocated to three groups (intravenous inoculation, G1; intradermal inoculation, G2; negative control, G3). G1 and G2 dogs were inoculated twice (day 0, day 28) with 10 8 promastigotes of L . tarentolae strain (RTAR/IT/21/RI-325) isolated from a Tarentola mauritanica gecko. The animals were followed until day 206. Blood, serum, conjunctival swabs and lymph node aspirate samples were collected monthly and bone marrow, liver and spleen biopsies on day 91. Hematological and biochemical parameters were assessed monthly, as well as serology (IFAT and ELISA) and molecular identification of L . tarentolae . Mononuclear cells (PBMC) were obtained to assess the cytokine expression through in vitro stimulation or (re-) infection. Data from this study demonstrated that DNA from L . tarentolae is detectable up to 3 months post-infection, with seroconversion after day 28. Moreover, the non-pathogenic nature of L . tarentolae was confirmed, with a neutral Th1/Th2 polarization, and a possible shift to Th1 phenotype after derived macrophages (re-) infection, as demonstrated by the expression of IFN-gamma. Therefore, L . tarentolae demonstrated a great potential as a surrogate pathogen and/or immune-prophylaxis/immune-therapy against Leishmania infections in dogs and humans.

During 2021, we collected blood and serum samples from 135 persons exposed to tick bites in southern Italy. We serologically and molecularly screened for zoonotic tickborne pathogens and only molecularly screened for Candidatus Midichloria mitochondrii. Overall, 62 (45.9%) persons tested positive for tickborne pathogens. Coxiella burnetii was detected most frequently (27.4%), along with Rickettsia spp. (21.5%) and Borrelia spp. (10.4%). We detected Candidatus M. mitochondrii DNA in 46 (34.1%) participants who had statistically significant associations to tickborne pathogens (p<0.0001). Phylogenetic analysis of Candidatus M. mitochondrii sequences revealed 5 clades and 8 human sequence types that correlated with vertebrates, Ixodes spp. ticks, and countries in Europe. These data demonstrated a high circulation of tickborne pathogens and Candidatus M. mitochondrii DNA in persons participating in outdoor activities in southern Italy. Our study shows how coordinated surveillance among patients, clinicians, and veterinarians could inform a One Health approach for monitoring and controlling the circulation of tickborne pathogens.

Background Canine leishmaniosis (CanL), a sand fly-borne zoonotic disease caused by Leishmania infantum , is potentially lethal in dogs. A similar or slightly higher quantity of antigens over antibodies promotes the formation of soluble circulating immune complexes (sCIC), which are deposited in the capillary wall, causing the inflammatory cascade responsible for clinical manifestations. Nervous system involvement during CanL is rarely reported in both humans and dogs, and the exact underlying process involving the peripheral nervous system (PNS) is still debated in both species. Methods Two male mixed-breed dogs were presented for exercise intolerance, non-ambulatory flaccid tetraparesis and decreased/absent flexor reflexes in all four limbs. Both dogs were seropositive for L. infantum and presented clinicopathological abnormalities suggestive of active CanL. One dog had received N -methyl-glucamine antimoniate two months before presentation without neurological improvement. Results Generalized PNS involvement was confirmed in both dogs. Biopsies of muscle and nerve tissues showed mononuclear cell inflammatory infiltration, and quantitative real-time polymerase chain reaction (PCR) was positive for Leishmania spp. In addition, Leishmania spp. antigen was detected in the nerve from one dog by immunohistochemistry. Both dogs were started on N -methyl-glucamine antimoniate and allopurinol in association with immunosuppressive corticosteroid therapy, recovering in few weeks. Conclusions Peripheral neuropathies during active CanL can be a consequence of sCIC deposition on endoneurial vascular endothelium comprising the blood–nerve barrier and its consequent breakdown. However, an abnormal host immune response triggered by L. infantum causing demyelination and/or axonal disruption is also possible. The positive response to the immunosuppressive therapy further supports an immune-mediated origin of the PNS condition. Therefore, CanL should be included in the differential diagnosis of PNS disease in dogs, especially in areas endemic for L. infantum . Graphical Abstract