New insights about Haplosporidium pinnae and the pen shell Pinna nobilis mass mortality events.

Since early autumn 2016, Mass Mortality Events (MME) have drastically impacted the population of the fan mussel Pinna nobilis in the Mediterranean Sea. Haplosporidium pinnae, a newly described Haplosporidian species, has been considered the causative agent of the mortality outbreaks in association to opportunistic bacterial pathogens. In the present study, we first reported a cytological description of H. pinnae in moribund specimens of P. nobilis which were collected in the Gulf of Taranto (Ionian Sea, Italy) during summer 2018. Different life-cycle stages of the parasite, including uni- and binucleate cells, small plasmodia, big multinucleate plasmodia and sporocysts with spores, were detected in all the examined animals and most of the parasite cells were present in gills, mantle and digestive gland, while the spores were found only in the latter organ. Histology and molecular biology were also performed, confirming the nature of the infectious agent, as already reported in the area. Additionally, molecular study revealed the presence of bacteria from the Mycobacterium ulcerans - M. marinum complex but no evident macroscopical or microscopical lesions, just as no bacteria referred to Mycobacterium were observed by histology. In conclusion, the present study aimed to provide further contributions to the understanding of the mortality of P. nobilis, pointing to the role of the cytological method of investigation both for diagnostic and epidemiological purposes, and discussing the current epidemic situation in the Adriatic sea.

Epidemiological survey on cystic echinococcosis in wild boar from Central Italy.

Despite the wide distribution of wild boar populations in Italy and the increase of its diffusion in urbanized areas, only one case report has described the occurrence of Echinococcus granulosus s.l. in a wild boar from Marche (Central Italy). The present study investigated the presence of E. granulosus sensu lato with an epidemiological survey on wild boars from central Italy that had been killed during hunting season. Seven hundred sixty-five (765) adult wild boars were examined during the 2016-2017 hunting season. Of these animals, 1.0% (8/765) were positive to E. granulosus s.l. with a fertility of 0.3% (2/765), and 2.9% animals (22/765) were positive for the metacestode stage of Taenia hydatigena (Cysticercus tenuicollis), while 0.5% (4/765) showed mixed infection (E. granulosus s.l. + T. hydatigena). Sixteen hydatids were found, of which 12.5% were fertile, 37.5% were sterile, 31.3% were caseous, and 18.8% were calcified. Eight hydatids (two fertile and six sterile cysts) were molecularly characterized by analysis of the mitochondrial gene, cytochrome c oxidase subunit 1 (cox1), and the NADH dehydrogenase subunit 1 (ND1). Hydatids found in wild boars were characterized as E. granulosus sensu stricto (G1 genotype). The present survey represents the first epidemiological study on cystic echinococcosis in wild boar in Italy which highlights the need for more extensive epidemiological investigations to determine the causal factors, economic impact, and public health importance of the disease in this livestock-wildlife setting.

Detection of selected pathogens in Apennine wolf (Canis lupus italicus) by a non-invasive GPS-based telemetry sampling of two packs from Majella National Park, Italy.

In this study, a multi-pathogens survey was conducted to verify the sanitary status of two Italian wolf packs of Majella National Park. Twenty fecal samples (10/pack) were collected using a sampling protocol, based on the combining data from radio-collared wolves with geographic information system (GIS) analysis, allowing to mark off the home range of packs and to recover group-specific and high-quality specimens. Virological screening against the most prevalent canine viruses (protoparvovirus, distemper virus, adenoviruses, and coronaviruses) was carried out by molecular methods, while parasites were detected by means of copromicroscopic and molecular analysis. Canine parvovirus type 2b (CPV-2b) is the most prevalent virus in both packs (7/20), followed by canine adenovirus type 2 (CAdV-2), while no sequences of canine distemper virus and coronaviruses were detected. The sequence analysis of the viruses demonstrated the domestic origin of the infection, highlighting the importance of vaccination of local dogs in order to reduce the risk of exposure of wildlife to these pathogens. Fourteen samples resulted positive for parasites. Capillaria aerophila (sin. Eucoleus aerophilus), Ancylostoma/Uncinaria, Trichuris vulpis eggs, Sarcocystis spp., Cystoisospora canis, and Angiostrongylus vasorum larvae were identified. Echinococcus granulosus sensu stricto (ovine genotype G1) and Giardia duodenalis(canid-specific Assemblage C) were also characterized, providing insights into the wolves' diet and their effects on environmental contamination. The sampling protocol applied in this study, based on a multidisciplinary approach, represents an innovative tool for the survey of Apennine wolf, able to integrate sanitary data with the ecological and demographic features of this population.

Zoonotic parasites in feces and fur of stray and private dogs from Italy.

The present study evaluated the occurrence of zoonotic parasites in feces and on fur of stray and private dogs living in Italy. Individual fecal samples collected from 117 and 385 kenneled (i.e., rescue shelter) and privately owned dogs, respectively, were examined by conventional copromicroscopy. Samples positive for Giardia duodenalis and Cryptosporidium spp. were subjected to further genetic characterization. Additionally, 174 fur samples of private and kenneled dogs living in the same area, 27 of which were also subjected to copromicroscopical examinations, were examined for the presence and viability of parasite elements. Out of 502 fecal samples, forty-one (8.2%) scored positive for G. duodenalis cysts while one (0.2%) for Cryptosporidium oocysts. Nine private dogs were molecularly positive for G. duodenalis assemblage C (n. 7), assemblage D (n. 1), and assemblages C + D (n. 1), while 11 kenneled dogs were positive for assemblage C (n. 8), assemblage D (n. 1), and assemblages C + D (n. 2). Cryptosporidium spp. were identified as C. canis in a private dog. One hundred and seventy-two dogs (34.3%), i.e., 92 private and 80 kenneled, were positive for zoonotic nematodes: Toxocara canis (6.6 %), Ancylostoma caninum (6.8%), and Capillaria aerophila (0.6%). Unviable eggs of Toxocara spp. were found on the hair of five (2.9%) private dogs. The present findings indicate that canine feces from both private and kenneled animals may contain zoonotic parasites and may be a potential risk for humans and other animals, especially when they contaminate the environment. The role of dog fur as source of human infections should be further investigated.

A pilot study of the in vitro efficacy of different concentrations of Duddingtonia flagrans for the control of gastrointestinal nematodes of sheep.

Duddingtonia flagrans is a nematode trapping fungus used for the control of gastrointestinal nematodes in livestock. The quantity of chlamydospores of D. flagrans required for the reduction of third-stage larvae (L3) of sheep gastrointestinal nematodes (GIN) is largely unknown, and a matter of discussion. The aim of this experiment was to determine in vitro the nematophagous activity of four different concentrations of D. flagrans (1000, 3000, 6250, or 11000 chlamydospores/ml) in the presence of varying numbers of GIN third-stage larvae (L3) (500, 1000, 1500). Additionally, the study sought to evaluate the efficacy of this fungus on Haemonchus contortus, Teladorsagia circumcincta, Trichostrongylus colubriformis and Chabertia ovina. The results showed that as fungal concentrations increased, so did the larval reduction of third-stage infective larvae in each test. L3s number was not a determining factor in the efficacy against GIN. The comparison between various concentrations of chlamydospores revealed significant differences, particularly between 1000 and 11000 chlamydospores (P≤0.05). Regarding the larval reduction of the GIN species considered, D. flagrans demonstrated the same effectiveness across all species tested. The results of the current study confirm the efficacy and underscore the importance of D. flagrans as an alternative for controlling of GIN.

Hepatozoon spp. in stray cats from the metropolitan area of Rio de Janeiro, Brazil.

In the last few years, the number of studies on feline hepatozoonosis has increased, but our knowledge on the actual species of Hepatozoon and/or different genotypes affecting felines is still incipient. At least three species, namely Hepatozoon felis, H. canis, and H. silvestris, have been isolated from domestic cats in various countries. Additionally, there are indications that other species and genotypes may affect felines in given geographic areas. This study was carried out to investigate the occurrence of Hepatozoon spp. in cats from Niterói, a municipality within the metropolitan area of Rio de Janeiro, Brazil. Individual blood samples were collected from 28 cats enrolled in a spaying/castration program. DNA was extracted from all samples and subjected to sequencing specific for Hepatozoon spp. DNA of H. felis was found in 21/28 cats (75%), and four genetic polymorphisms never described thus far were detected. This is the first report of H. felis in cats living in the State of Rio de Janeiro, and the present data confirm that H. felis is a species complex encompassing different genotypes circulating within cat populations. Further studies are warranted to investigate whether different genotypes have different biology or pathogenicity for felids.

Title: Hepatozoon spp. chez les chats errants de la zone métropolitaine de Rio de Janeiro, Brésil. Abstract: Au cours des dernières années, le nombre d'études sur l'hépatozoonose féline a augmenté, mais nos connaissances sur les espèces d'Hepatozoon et/ou différents génotypes affectant les félins sont encore naissantes. Au moins trois espèces, à savoir Hepatozoon felis, H. canis et H. silvestris, ont été isolées chez des chats domestiques dans divers pays. De plus, il semble que d'autres espèces et génotypes puissent affecter les félins dans des zones géographiques données. Cette étude a été réalisée pour étudier la présence d'Hepatozoon spp. chez des chats de Niterói, une municipalité de la zone métropolitaine de Rio de Janeiro, au Brésil. Des échantillons de sang ont été prélevés individuellement sur 28 chats d'un programme de castration. L'ADN a été extrait de tous les échantillons et soumis à un séquençage spécifique de Hepatozoon spp. L'ADN de H. felis a été trouvé chez 21 chats sur 28 (75%) et quatre polymorphismes génétiques, jamais décrits jusqu'à présent, ont été détectés. Il s'agit du premier signalement de H. felis chez des chats vivant dans l'État de Rio de Janeiro et les données actuelles confirment que H. felis est un complexe d'espèces englobant différents génotypes circulant au sein des populations de chats. Des études supplémentaires sont nécessaires pour déterminer si les différents génotypes ont une biologie ou une pathogénicité différente pour les félidés.

Comparison of diagnostic methods for laboratory diagnosis of the zoonotic tapeworm Dipylidium caninum in cats.

The tapeworm Dipylidium caninum is the most widely distributed cestode infecting dogs, cats, and sometimes humans, worldwide. The diagnosis of the infection caused by D. caninum is achieved via the visualization of proglottids in feces or with traditional microscopic tests, but both lack sensitivity. The present study has evaluated and compared the diagnostic performance of a PCR protocol on different feline biological samples to detect D. caninum. A sample of feces, a Scotch tape test from the perianal area, and a rectal swab were collected from a total of 100 privately owned cats from Italy and Greece. All fecal samples were subjected to macroscopic examination and to floatation. Based on the results of the above tests the cats were divided in three groups, i.e. (i) cats positive for D. caninum (regardless of positivity for other endoparasites (Group A; n = 50 cats), (ii) cats negative for D. caninum but infected by other helminths (Group B; n = 25 cats), and (iii) cats negative for intestinal endoparasites (Group C; n = 25 cats). For each sample, the DNA was extracted from feces, floatation supernatant, Scotch tape test and rectal swabs and subjected to PCR. For 33 cats from Group A, at least one sample type scored positive at PCR. Of these, all were PCR-positive in the floatation aliquot, while nine and one cats were positive by PCR on feces and Scotch tape test, respectively. Swabs were negative by PCR for all the cats. None of the samples from cats of Groups B and C was positive by any PCR. Sequences obtained from amplicons generated from samples of cats enrolled in Italy had 99-100 % identity with the recently described D. caninum feline genotype. The data presented here suggest that PCR could be a useful tool for diagnosing D. caninum infections, under certain circumstances, e.g. when proglottids are unidentified, unseen or overlooked, even though it has limitations, e.g. false negative results due to fecal PCR inhibitors, uneven distribution of parasitic elements, or to intermittent proglottid and/or egg shedding. Thus, it may not be, currently, the best diagnostic choice for dipylidiosis.

Geographical isolation and hyperendemicity of Hepatozoon felis: Epidemiological scenario in Skopelos, Greece, and phylogenetic analysis.

Feline hepatozoonosis is a vector-borne disease caused by different species of the genus Hepatozoon, i.e. Hepatozoon felis, Hepatozoon silvestris and Hepatozoon canis. Knowledge on the biology, epidemiology and taxonomy of Hepatozoon spp. is still limited, despite the fact that the number of documented Hepatozoon spp. infections in domestic cats increased in recent years in different countries. This study was carried out to evaluate the prevalence and the genetic profile of Hepatozoon spp. in cats living on the island of Skopelos, Greece. Individual blood samples were collected from 54 owned cats and were subjected to Giemsa-stained blood smear examination to investigate the presence of Hepatozoon spp. gamonts and to a specific PCR protocol targeting the 18S rRNA gene of Hepatozoon. A total of 45 cats (83.3%) were found infected by Hepatozoon spp. by at least one of the methods applied. In particular, 43 (79.6%) of the cats were PCR-positive, and in 6 (11.1%) cats gamonts of Hepatozoon spp. were found in the blood smears. A total of 26 H. felis sequences were obtained and the presence of three undescribed single nucleotide polymorphisms were detected. The present results indicate that H. felis species complex may be hyperendemic in isolated/confined areas. In such contexts, geographical isolation may favor the origin of new genotypes or haplotypes or even new species.

Molecular detection of Capillaria aerophila, an agent of canine and feline pulmonary capillariosis.

Capillaria aerophila, a trichuroid nematode causing pulmonary infections in wild and domestic carnivores, is occasionally and potentially poorly recognized in infections of humans due to clinicopathological mimicry and a lack of accurate, robust laboratory diagnostics. The present work evaluated the efficiency of a DNA-based assay amplifying a partial cytochrome c oxidase subunit 1 (cox1) gene of C. aerophila in the diagnosis of lung capillariosis. Fecal samples from 34 dogs and 10 cats positive at parasitological examination for C. aerophila and other endoparasites (i.e., other lungworms, whipworms, roundworms, hookworms, tapeworms, and/or coccidia) and from 44 animals negative for C. aerophila but positive for other endoparasites were molecularly examined. Of the 44 samples positive for C. aerophila at copromicroscopy, 43 scored positive (i.e., 33/34 dogs and 10/10 cats) in seminested PCR, resulting in a sensitivity of 97 to 100%. Samples that were copromicroscopy negative for C. aerophila although positive for other endoparasites never produced a PCR product or nonspecific amplicons. The specific PCR amplification of C. aerophila (i.e., specificity of 100%) was confirmed by a nucleotide sequence analysis of the cox1 amplicons. The potential implications of the molecular diagnosis of lung capillariosis are discussed.

Animal Welfare and Parasite Infections in Organic and Conventional Dairy Farms: A Comparative Pilot Study in Central Italy.

The study investigated and compared welfare conditions and gastrointestinal (GI) parasites distribution among organic (ORG) and conventional (CONV) farms in central Italy. Five ORG and five CONV farms were assessed for animal welfare with an adapted version of the AssureWel protocol. Faecal samples collected from the rectum of the animals both in ORG (n = 150) and CONV (n = 150) were analysed using conventional copromiscroscopy. The presence of skin damages in the rear legs was significantly predominant (p < 0.001) in CONV (26.7%) compared with ORG farms (10.0%). No differences were found for lameness, cleanliness, Body Condition Score, hair loss, body lesions and swelling prevalence. Data concerning the productive performances, e.g., total milk, fat and protein yields standardised in mature equivalent (ME) were collected. ME milk yield (ORG: 9656.9 ± 1620.7 kg; CONV: 12,047.2 ± 2635.3) and ME fat yield (ORG: 396.6 ± 66.8; CONV: 450.3 ± 102.8) were significantly lower in ORG farms (p < 0.001). Anthelmintics were used regularly in 4/5 CONV and 0/5 ORG farms. In 2 CONV farms (40%) and 4 ORGs (80%) at least one animal tested positive for GI parasites. No significant differences in parasites prevalence emerged (ORG = 10.7%; CONV = 8%). These data indicate that ORG farming does not influence parasite prevalence and animal welfare status.

An Uncommon and Severe Clinical Case of Sarcoptes scabiei Infestation in a Cat.

The burrowing mite Sarcoptes scabiei causes sarcoptic mange in dogs and other mammals, including humans. Despite S. scabiei infests several animals, little is known about the epidemiology, clinical features, and treatment of sarcoptic mange in cats. A few reports have shown that clinical signs of S. scabiei infestation in cats may vary from non-pruritic crusted lesions to itchy and mild lesions, while severe signs are very infrequent. The present report describes a severe case of S. scabiei infestation in a stray cat from Italy, characterized by bilateral alopecia, diffuse and multifocal hyperpigmented, crusted, erythematous, scaled, and exfoliated lesions. The cat was FIV-positive and also infected by the cestode Dipylidium caninum. After treatment with a broad-spectrum parasiticide and an antibiotic, the cat showed an almost complete clinical recovery in 4 weeks. Unfortunately, no further clinical examinations were performed due to the lack of compliance of the owner and to the death of the cat for causes unrelated to sarcoptic mange. This clinical case indicates that under certain circumstances, S. scabiei can reproduce and cause severe clinical signs in cats which are usually considered non-permissive hosts for this mite, and in which the disease is usually self-limiting. Clinic-pathological, epidemiological, and zoonotic implications are discussed.

Ticks, Fleas, and Harboured Pathogens from Dogs and Cats in Cyprus.

Ticks and fleas are blood-sucking ectoparasites that cause irritation and anaemia to their hosts and act as vectors of pathogens (vector-borne pathogens, VBPs) of relevance for animal and human health. In the present study, tick and flea species in dogs and cats from Cyprus were recorded and VBPs were detected in the collected specimens. Ectoparasites were collected from 220 animals (161 dogs and 59 cats), and a questionnaire including demographic, clinical, and other information was filled out for each animal. The ectoparasites were morphologically identified and the detection of VBPs was performed by PCR-coupled sequencing. Rhipicephalus sanguineus sensu lato was found on 108 dogs and 13 cats, and Ixodes gibbosus on 2 dogs. Ctenocephalides felis was the predominant flea species (on 62 dogs and 45 cats), while one dog and one cat were infested by Ctenocephalides canis and Echidnophaga gallinacea, respectively. The VBPs in ticks were Anaplasma platys, Rickettsia massiliae, Rickettsia conorii, Rickettsia felis, Hepatozoon felis and Hepatozoon canis, while Rickettsia felis, Rickettsia sp., Bartonella koehlerae, Bartonella clarridgeiae, and Bartonella henselae were recorded in fleas. Statistical analysis (chi-square test and multiple univariate generalized linear model) showed that animals up to 6 months of age were less likely to be infested with ticks than older animals, but more likely to be infested with fleas. Ticks were more prevalent in sheltered than in owned animals, while the odds ratio of flea presence was higher in owned animals than those living in shelters. The present study is the first investigation on the occurrence of ticks and fleas in dogs and cats from Cyprus, showing the presence of different VBPs in these important ectoparasites. The results point out the importance of systematic ectoparasite control in dogs and cats.

Zoonotic intestinal helminthes diagnosed in a 6-year period (2015-2020) in privately owned dogs of sub-urban and urban areas of Italy.

Intestinal helminthes of dogs have a relevant role for health and welfare of canine populations and many of them are zoonotic. Thus, a constant surveillance is pivotal to safeguard companion dogs and human health. Data from faecal examinations of pet dogs obtained in the last six years during the routine activity of the laboratory of Parasitology and Parasitic diseases of the University Teaching Veterinary Hospital of the University of Teramo, Italy, are here reported. From 2015 to 2020 a total of 1083 individual faecal samples from privately owned dogs were examined using conventional copro-microscopic techniques. Endoparasite elements were found in 278 (25.7%) dogs, and among them, eggs of intestinal zoonotic helminthes were detected in 184 (17%) samples. The most common zoonotic parasites were Toxocara canis and hookworms (8.9% positivity rate for both). Most dogs (88%) were positive for a single zoonotic parasite while 12% of the positive animals harbored two or more zoonotic helminthes. A statistical analysis showed a significant association between dogs with less than one year of age and positivity to at least one intestinal zoonotic helminth, and that infections by T. canis are significantly more prevalent in dogs under one year of age. Despite being well known in canine clinical practice, these results indicate that zoonotic helminthes are still frequent in owned dogs. Thus, increased awareness of veterinary practitioners and pet owners is necessary to implement current diagnostic and control strategies to minimize the risk of zoonotic transmission of intestinal helminthes.

A Comparison of Copromicroscopic and Molecular Methods for the Diagnosis of Cat Aelurostrongylosis.

The gold standard method for the diagnosis of cat aelurostrongylosis is the detection of Aelurostrongylus abstrusus first stage larvae with the Baermann's examination. Nevertheless, molecular assays have shown higher diagnostic performances compared to copromicroscopy. This study evaluated the usefulness of an A. abstrusus species-specific PCR on different biological samples collected in clinical settings from 100 privately-owned cats in Italy (n. 60) and Greece (n. 40). A fecal sample was collected from each animal and a pharyngeal swab was also obtained for cats from Italy. All stool samples were subjected to flotation and Baermann's test. The cats were categorized in three groups based on the results of copromicroscopy, i.e., Group A (n. 50 cats with A. abstrusus infection regardless of positivity for other helminths), Group B (n. 25 cats negative for A. abstrusus but positive for at least one of any other helminth), Group C (n. 25 cats negative for any helminth). DNA was extracted from individual aliquots of feces, flotation supernatant, Baermann's sediment and the pharyngeal swab and then subjected to a PCR specific for A. abstrusus. At least one fecal aliquot or the pharyngeal swab scored positive by the A. abstrusus-specific PCR for 48/50 (96%) cats enrolled in Group A; in particular, 38/50 (76%), 35/50 (70%), 41/50 (82%) and 21/25 (84%) DNA extracts from feces, flotation supernatant, Baermann's sediment and pharyngeal swabs were positive by PCR. These results confirm that molecular tools are highly sensitive and specific and indicate that pharyngeal swabs are the most suitable sample for molecular analysis in clinical settings.

Parasitic cysts in wild boars hunted in Central Italy: The sanitary controls in the wild game meats chain.

Seven hundred sixty-five (765) adult wild boars were examined during the 2016/2017 hunting season for the research of parasites. Post mortem inspection was carried out at the slaughterhouse by the Official Veterinarian on the plucks (heart, tongue, lungs, diaphragm, and liver) of the killed animals presented by hunters. Of these, 0.8% (6/765) were positive for Echinococcus granulosus sensu lato (s.l.), and 2.6% (20/765) were positive for the metacestode stage of Taenia hydatigena (Cysticercus tenuicollis), while 0.5% (4/765) animals showed a mixed infection (Echinococcus granulosus s.l. and Taenia hydatigena). Sixty-three (63) cystic lesions were found. Of these 25,4% (16/63) were caused by Echinococcus granulosus s.l. and 74,6% (47/63) were caused by Cysticercus tenuicollis. The more involved organs were liver and lungs, in a less extension omentum and diaphragm. Parasitological analyses showed an overall prevalence of 3.9% for metacestodes in the hunted animals examined (Paoletti et al., 2018). Hydatids were molecularly characterized as E. granulosus sensu stricto. Trichinella spp. examination results showed no evidences of parasitic cysts. The products of hunting used for own consumption and direct sale to the final consumer or retailer, according to Regulation (EC) No 853/2004, lack of overall control by the Competent Authority. This is a critical point in the food chain of the game meat. The data obtained show the importance of the post mortem inspection and the central role of the Competent Authority to ensure not only the food safety of game meat but also to collect data for extensive epidemiological investigations on live-stocks wildlife settings having a direct impact on public health.

Tritrichomonas foetus in cats from Central Italy: Clinical signs and risk factors.

Tritrichomonas foetus (Trichomonadida, Tritrichomonadidae) is a parasitic protozoan which causes a diarrheic disease in domestic cats worldwide. The epizootiological and pathogenic impact of this parasite in many geographic locations is still to be unraveled; thus, the present study has investigated the occurrence of T. foetus in cats from central Italy, along with the analysis of risk factors and clinical signs. One hundred and five cats referred to the University Veterinary Teaching Hospital of the University of Teramo were tested with copromicroscopic techniques for common intestinal and extra-intestinal parasites and a specific PCR for T. foetus. A statistical analysis evaluating associations between T. foetus positivity and possible risk factors was performed. One animal scored positive for Giardia (0.95%), 2 for Toxocara cati (1.90%), 1 for Eucoleus aerophilus (syn. Capillaria aerophila) (0.95%), 9 for Aelurostrongylus abstrusus (8.57%) and 1 for Dipylididum caninum (0.95%), while T. foetus was molecularly recorded in 18/105 (17.14%) of the study cats. In two cats, T. foetus was found in association respectively with Giardia and A. abstrusus. Animals with chronic gastrointestinal signs were more significantly infected by T. foetus (40.90% vs 10.84%; p < 0.01; 95% CI = 1.90-17.04; OR = 5.69) in comparison to cats with no gastrointestinal distresses. No other statistically significant associations were found. This study confirms that T. foetus is enzootic in cats in Italy and that it should be included in the differential diagnosis of feline chronic gastrointestinal diseases.

First record of Hepatozoon spp. in domestic cats in Greece.

Feline hepatozoonosis is an emerging disease of domestic and wild felids though there is limited knowledge of this infection, e.g. regarding geographical distribution and parasite species involved. The present study evaluated microscopically and molecularly the occurrence of Hepatozoon spp. in domestic cats from insular (Crete, Mykonos and Skopelos) and continental (prefectures of Attica and Thessaloniki) Greece. Out of 282 cats examined, 72 (25.5 %) scored positive by PCR for Hepatozoon spp. and of them, 9 (12.5 %) showed gamonts on the blood smear microscopic examination. Sequences obtained from 35 of the amplicons proved the presence of two haplotypes of Hepatozoon felis. One, herein called H1 (34/35 amplicons) resulted 100 % identical with H. felis from Italy and isolates from other continents, and ∼98 % similar with a H. felis isolate causing severe clinical signs in Austria. The haplotype H2, found in a cat in Skopelos, had ∼94 % identity with H1, with H. felis isolates from Italy, Israel, Spain, a ∼92 % identity with the isolate from Austria, and ∼94-98 % with isolates from South Africa. These are the first records of H. felis in cat populations from Greece and indicate that the infection may be present at high prevalences in different regions of the country. Furthermore, the results of the molecular and phylogenetic analysis support a recent hypothesis indicating the existence of a species-complex classification for H. felis. Further studies aiming at elucidating the genetic make-up of Hepatozoon populations and possible variations in terms of geographic distribution and clinical relevance are necessary. The importance of a continuous epizootiological monitoring is crucial for the establishment of preventative and control measures protecting the health of cats living in or travelling to enzootic areas.

Leishmania infantum Seroprevalence in Cats From Touristic Areas of Italy and Greece.

Leishmaniosis by Leishmania infantum is a major zoonotic Vector-Borne Disease (VBD) in terms of geographic distribution, pathogenicity and zoonotic potential. While dogs are the main reservoir of L. infantum, the infection in cats is poorly understood although increasingly reported from enzootic and non-enzootic areas. The Mediterranean basin is a key area for leishmaniosis and includes touristic spots that require continuous surveillance for VBDs in consideration of the growing tendency of tourists to travel with their pets. This study evaluated L. infantum seroprevalence in cats living in selected touristic localities of Italy and Greece. A total of 269 cat serum samples from three Sites i.e., 76, 40, and 153 from Adriatic Coast of Abruzzo, Italy (Site A), Giglio Island, Tuscany, Italy (Site B), and Mykonos Island, Greece (Site C), respectively, were included in the survey. Sera samples were subjected to an indirect immunofluorescence antibody assay for the detection of anti-L. infantum specific IgG. Associations between possible risk factors and seropositivity to L. infantum were statistically evaluated. Antibodies against L. infantum were detected in eight out of 269 (3.0%) cats tested i.e., 4/76 (5.3%), 1/40 (2.5%), and 3/153 (2.0%), from sites A, B, and C, respectively. A statistical association between anti-L. infantum antibodies and cohabitation with dogs was shown. This study indicates that feline populations living in the examined Italian and Greek touristic areas are exposed to L. infantum and that they may contribute to the circulation of L. infantum, enhancing the risk of infection for dogs and humans.

Immunoenzymatic analysis and genetic detection of Cryptosporidium parvum in lambs from Italy.

Cryptosporidiosis is a worldwide-diffused protozoan disease causing important economic losses to animal husbandry and livestock production. Additionally, several species/genotypes of Cryptosporidium have a relevant zoonotic potential and ruminants may be important sources of infection for human beings. Nonetheless, in Europe, little is known of the presence of Cryptosporidium in sheep nor on the species/genotypes involved. To obtain information on the occurrence of cryptosporidiosis in lambs and the potential zoonotic role of the Cryptosporidium isolates, one hundred and forty-nine faecal samples individually collected from lambs in central Italy have been examined for the presence of Cryptosporidium. All faecal specimens were processed with a commercial ELISA kit immunoassay and all ELISA-positive samples were further analyzed genetically. Twenty-six ELISA-positive samples scored positive at the PCR and the sequences obtained displayed 100% identity with the zoonotic Cryptosporidum parvum. This work suggests for the first time that lambs in Italy may shed C. parvum, thus representing a potential public health hazard.