Clinical and Clinico-Pathological Observations of the Erythrocyte Sedimentation Rate in Dogs Affected by Leishmaniosis and Other Inflammatory Diseases.

The erythrocyte sedimentation rate (ESR) has been used in canine medicine in several disorders, above all, to evaluate levels of inflammation. This study evaluated the ESR in canine leishmaniosis (CanL) and other inflammatory conditions. Three groups of dogs were examined: CanL affected dogs without clinical signs (INFECTED group, #25) or with clinical signs (SICK group, #43) and dogs affected by acute or acute-on-chronic conditions (OTHER DISEASE group, #65). The ESR was compared with acute phase proteins or reactants either positive or negative (leukogram, fibrinogen, iron, unsaturated iron binding capacity, ferritin, haptoglobin, and albumin) and immunological markers (gamma-globulins, IgG, and IgM). The ESR was higher in the SICK group than in the INFECTED group (median 39 vs. 11 mm/h; p < 0.0001), as well as in the OTHER DISEASE than in the INFECTED groups (median 41 vs. 11 mm/h; p < 0.0001). The ESR appeared outside the reference range for all dogs in the SICK and OTHER DISEASE groups and almost with similar values (mm/h; median 39, 95% CI 31-51 vs. 41, 95% CI 12-87; p > 0.05). The extent of changes in ESR can help to establish the severity of CanL and other inflammatory disorders. As a point-of-care test, the ESR can be used to screen dogs for unhealthy conditions, and its values correlate with the severity of any disease, including CanL.

Linguatula serrata (Pentastomida: Linguatulidae) infection in a paucisymptomatic greyhound imported from Romania to Italy: A case report and literature overview.

Background: Linguatula serrata is a pentastomid zoonotic parasite with worldwide distribution. Although some cases of L. serrata infection have been reported in dogs, the epidemiology of this parasite remains largely unknown in developed countries. In recent years, canine linguatulosis has been repeatedly linked to cases of imported infections. This study aims to focus attention on this uncommon parasite through the presentation of a case report and an overview of the literature. Case Description: A 1-year-old intact female Borzoi imported from Romania to Italy sneezed spontaneously a worm-like parasite specimen. Morphological and molecular diagnosis identified the parasite as a female of the zoonotic pentastomid L. serrata (so-called European tongue worm) that lives in the nasopharyngeal tract of canids. Eggs of Linguatula were detected in the feces. Molecular identification (99%-100% homology) was based on DNA extraction, polymerase chain reaction of a 700-bp fragment of the mitochondrial cytochrome c oxidase subunit I gene, and alignment with BLAST analysis. Due to the possibility of other adult or juvenile specimens of the parasite still occurring in the dog, a treatment attempt with a combination of febantel/pyrantel/praziquantel was made. No parasite eggs were detected in fecal samples after the drug was administered. Endoscopy confirmed the absence of adult parasites and slight pathological changes. A follow-up examination conducted 3 months after the treatment did not reveal any clinical and laboratory abnormalities. Conclusion: Linguatula serrata appears to be currently prevalent in some European countries, but there are no recent extensive studies on the prevalence of canine linguatulosis, so the parasite frequently remains undetected and unreported in dogs as the diagnosis is often overlooked. Parasites not commonly found such as L. serrata can become increasingly prevalent and may be detected in imported dogs. Therefore, veterinarians must be aware of the possible presence of uncommon and exotic pathogens in these dogs, be able to recognize the relevant clinical signs, and diagnose the infection quickly. This will improve the prognosis in individual dogs, reduces the risk of possible public health implications, and reduces the risk of uncommon and exotic pathogens establishing new endemic foci.

A One-Year Retrospective Analysis of Viral and Parasitological Agents in Wildlife Animals Admitted to a First Aid Hospital.

This study aimed to provide information on the presence and frequency of viral and parasitic agents in wildlife presented to a Veterinary Teaching Hospital in 2020-2021. Serum and faecal samples were collected from 50 rescued animals (roe deer, fallow deer, foxes, badgers, pine martens, and porcupines) and examined by serological, molecular, and parasitological techniques. Transtracheal wash (TTW) was also collected post-mortem from roe deer. Overall, the results of the different techniques showed infections with the following viral and parasitic agents: Bovine Viral Diarrhea Virus, Small Ruminant Lentiviruses, Kobuvirus, Astrovirus, Canine Adenovirus 1, Bopivirus, gastrointestinal strongyles, Capillaria, Ancylostomatidae, Toxocara canis, Trichuris vulpis, Hymenolepis, Strongyloides, Eimeria, Isospora, Dictyocaulus, Angiostrongylus vasorum, Crenosoma, Dirofilaria immitis, Neospora caninum, Giardia duodenalis, and Cryptosporidium. Sequencing (Tpi locus) identified G. duodenalis sub-assemblages AI and BIV in one roe deer and one porcupine, respectively. Adult lungworms collected from the TTW were identified as Dictyocaulus capreolus (COX1 gene). This is the first molecular identification of G. duodenalis sub-assemblage AI and D. capreolus in roe deer in Italy. These results show a wide presence of pathogens in wild populations and provide an overview of environmental health surveillance.

What's in a child's ear? A case of otomyiasis by Sarcophaga argyrostoma (Diptera, Sarcophagidae).

A clinical report of otomyiasis in a 1-year-old girl is reported. A III instar larva of Sarcophaga sp. was microscopically identified and Sarcophaga (Liopygia) argyrostoma (Diptera, Sarcophagidae) was suspected. A molecular method targeting a fragment of the cox1 gene was used to confirm the identity of the specimen. Although myiases are not frequent manifestations in otolaryngology, they should arouse the attention of doctors, social workers and parents dealing with disabled people, the elderly and children. This contribution also highlights the need of combining microscopy and molecular tools to achieve a correct and reliable identification of the specimen/s.

Incidental Detection of Onchocerca Microfilariae in Donkeys (Equus asinus) in Italy: Report of Four Cases.

This paper reports the occurrence for the first time in Italy of autochthonous Onchocerca infection in donkeys. Four jennies, bred on the same farm, were referred to the Veterinary Teaching Hospital of Pisa for a check-up on ovarian activity (n = 3) or for veterinary support during the delivery (n = 1). Microfilariae were incidentally detected during the blood smear examination of one jenny. Peripheral blood samples were then collected from the other three jennies and the presence of microfilariae was investigated by Knott's test. Circulating unsheathed microfilariae were identified in all the animals. The level of microfilaraemia was between 1 and 31 microfilariae in 2 mL of blood. Hematological changes showed moderate eosinophilia in one case or both remarkable eosinophilia and basophilia in another case. Based on molecular findings by PCR and sequencing, the microfilariae showed 98% sequence similarity with Onchocerca sp. in the NCBI GenBank database (Accession No.: MK541848.1). The present report provides evidence that Onchocerca is an etiological agent of parasitic infection in donkeys in Italy. Our findings highlight the importance of screening in donkeys for Onchocerca even in the absence of clinical indications.

Wildlife Ungulate Rescue and Emergency Services in the Pisa Area (Tuscany, Italy): Evaluation of a 9-Years Period (2010-2018).

Background: We analyzed the clinical data of wildlife ungulates admitted for emergency care to the Veterinary Teaching Hospital (VTH), Department of Veterinary Medicine, University of Pisa over a 9-years period. Methods: Clinical data of all the wildlife ungulates admitted to the VTH were recorded. Blood samples were also taken from the animals for hematological and biochemical analysis. An assessment of ecto- and endoparasites was carried out, diagnostic imaging assessment was performed, and the outcomes were recorded. Results: Data concerning clinical parameters, blood work, parasitological analysis, and diagnostic imaging diagnosis were expressed as prevalence. Conclusion: The rescue and emergency treatments were related mostly to traumas caused by car accidents, followed by other causes. The traumatic injuries were mostly severe, characterized by multiple lesions involving hard and soft tissues. In this study, traffic accidents were the main cause of wildlife rescue and emergency management. This is probably due to the increased population of ungulates over the years, along with the considerable anthropization of the Pisa area.

Habronematidosis in Equids: Current Status, Advances, Future Challenges.

Over the past few decades, among equine parasitoses caused by gastrointestinal nematodes, habronematidosis has been discontinuously studied worldwide. Habronematidosis is a parasitic disease distributed all over the world. It is caused by Habronema microstoma, Habronema muscae, and Draschia megastoma (Spirurida, Habronematidae), and it is maintained in the environment by muscid flies which act as intermediate hosts. At larval and adult stages these species live in the stomach of domestic and wild equids. However, the larvae can also be found on the skin, causing lesions known as "summer sores", and occasionally on other body areas, such as ocular and genital mucosa (muco-cutaneous habronematidosis) and lung, liver, brain parenchyma. Depending on the parasite's developmental stage and localization site, clinical signs vary from mild to severe. Habronematidosis is responsible for significant economic losses, mostly when sport horses are affected, because their performances are impaired and the infection can be unaesthetic. We used three on-line databases for searching the articles on habronematidosis according to the selected inclusion criteria; a total of 250 contributions, published between 1911 and 2020 were analyzed. This review summarizes the key features of pathogenesis, epizootiology, diagnosis, treatment, and control of habronematidosis, and highlights the current knownledge about its geographical distribution and spread. Anthelmintic drugs are the most widely-used tools against habronematidosis; given the known risk of anthelmintic resistance in some nematodes affecting horses, this aspect should also be explored for habronematidosis. Dedicated research is essential to fill gaps of knowledge and increase the understanding of habronematidosis to maximize equine health, reduce economic losses and sanitary impact associated with this parasitic infection.

First multicenter coprological survey on helminth parasite communities of free-living loggerhead sea turtles Caretta caretta (Linnaeus, 1758) from the Adriatic Sea and Northern Ionian Sea.

The prevalence of endoparasite infections in 83 free-living specimens of Caretta caretta, classified as vulnerable species, from the Adriatic Sea and Northern Ionian Sea was investigated by coprological examination. Thirty-seven (44.6%) turtles were found to be infected with helminths. The helminth infections found were: Rhytidodes gelatinosus and Sulcascaris sulcata (18.1% each), Hapalotrema mistroides (13.2%), Cymatocarpus solearis (9.6%), Eniodotrema megachondrus (7.2%), Kathlania/Tonaudia sp. (3.6%), Neospirorchis sp., Octangium sagitta and Plesiochorus cymbiformis (1.2% each). There were no significant differences in the total prevalence of helminth infections between sexes, size classes, and seasonal periods. Conversely, the prevalence of helminth infections was significantly higher (P < 0.01) in accidentally caught turtles than in stranded turtles. Highly significant differences in prevalence of helminthiases were also seen among marine sampling areas. This report provides important baseline information about the helminth fauna of free-living C. caretta in the examined geographical region. This is also the first report of O. sagitta infection in C. caretta thus broadening the host range of the parasite.

Corrigendum: Wildlife Ungulate Rescue and Emergency Services in the Pisa Area (Tuscany, Italy): Evaluation of a 9-Years Period (2010-2018).

[This corrects the article DOI: 10.3389/fvets.2020.00626.].

Toxoplasma gondii and Other Zoonotic Protozoans in Mediterranean Mussel ( Mytilus galloprovincialis) and Blue Mussel ( Mytilus edulis): A Food Safety Concern?

Mediterranean mussels ( Mytilus galloprovincialis) and blue mussels ( Mytilus edulis) are among the most consumed fishery products, but they are frequent vehicles of foodborne infection worldwide. In this study, we investigated the occurrence and seasonality of zoonotic protozoans in mussels farmed or sold at retail outlets in Italy. We collected and tested 1,440 M. galloprovincialis and 180 M. edulis. Pooled samples were molecularly tested for Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii and then sequenced. Sixty-two (45.9%; 95% confidence interval, 37.5 to 54.3%) mussel pools tested positive for one or more of the investigated pathogens. Both Mytilus species and samples from all the investigated areas harbored pathogens. Mussels were statistically more contaminated by Cryptosporidium spp., followed by T. gondii and G. duodenalis assemblage A, and M. galloprovincialis was more contaminated than M. edulis ( P < 0.01). Contamination was more likely in mussels at retail outlets ( P < 0.05) than in those from farms and in mussels collected in spring ( P < 0.01) than in other seasons. This is the first report of T. gondii found in M. galloprovincialis in Italy and in M. edulis in Europe. The detection of zoonotic protozoans in a widely consumed food source indicates the need for a more detailed microbiological risk analysis, especially considering that bivalve mollusks are often consumed raw worldwide.

Toxoplasma gondii Lineages Circulating in Slaughtered Industrial Pigs and Potential Risk for Consumers.

Toxoplasma gondii is a cosmopolitan zoonotic protozoan parasite, and the consumption of raw or undercooked pig meat is one of the most important sources of T. gondii infection. Three predominant lineages, types I, II, and III, are widespread in Europe. Although still poorly understood, a relationship between each type and the severity of illness represents a public health issue. To gain further knowledge of the genotypes in circulation and of the potential risk for consumers, one heart sample and one diaphragm sample (206 total) were taken from each of 103 pig carcasses at an abattoir in Italy. Then, we used 529-bp repetitive element PCR and a B1 real-time PCR high-resolution melting assay coupled with sequencing to detect and genotype T. gondii isolates. T. gondii DNA was detected in 14 pigs (13.6%, 95% confidence interval = 7 to 20.2%), and types I (3.9%), II (5.8%), and III (3.9%) were identified. We found that heart tissue had a significantly higher PCR positivity rate for T. gondii than did diaphragm tissue. This is Europe's largest study on genotyping of T. gondii from pigs, and it demonstrates that all three main lineages are present in carcasses of industrially reared pigs in Italy. There is a potential risk to consumers of infection with any or all of the three lineages, and the related clinical consequences should be taken into account. This study suggests that monitoring of T. gondii types in meat is essential, especially in meat that is traditionally eaten raw or that is minimally processed.

Activity of selected essential oils on spoiling fungi cultured from Marzolino cheese.

Microscopic fungi can be present on a variety of foodstuff, including cheese. They can be responsible for fungal spoilage, causing sensory changes making food unacceptable for human consumption, and posing severe health concerns. Furthermore, some of these organisms are able to resist antimicrobial preservatives provided for by law. Antifungal activity of 15 chemically defined EOs, alone and in mixture, were checked by a microdilution test against isolates of Penicillium funiculosum and Mucor racemosus cultured from rinds of Marzolino, a typical Italian fresh pecorino cheese. Origanum vulgare yielded the lowest MIC values, followed by Salvia sclarea, Ocimum basilicum and Cymbopogon citratus, while Citrus paradisi and Citrus limon were not active. All mixtures showed antifungal activity at lower concentration with respect to MIC values of each EO component, when not in combination. This study is the first to describe the setting up of EOs mixtures to limit spoiling moulds.

Stonebrood and chalkbrood in Apis mellifera causing fungi: in vitro sensitivity to some essential oils.

Aim of the present study was to evaluate the in vitro antimycotic activity of 17 chemically defined essential oils (EOs) both alone and as a mixture, against agents responsible for stonebrood caused by Aspergillus flavus, and chalkbrood caused by Ascosphaera apis in European honeybees. Cinnamomum zeylanicum yielded the lowest MIC value against A. flavus, but was not effective against A. apis, while Litsea cubeba and Pelargonium graveolens appeared to be effective against all checked fungi. Aspergillus niger showed the lower sensitivity. Two mixtures composed by L. cubeba, C. zeylanicum and Cymbopogon flexuosus (M1) and by L. cubeba, C. zeylanicum, P. graveolens and C. flexuosus (M2), respectively, were tested, both resulting effective. The components of M1 showed a synergistic effect. The use of mixtures allowed to decrease the total amount of EOs. The use of these products could be of interest for an alternative natural approach in honeybee disease management.

Prevalence of the microsporidian Nosema ceranae in honeybee (Apis mellifera) apiaries in Central Italy.

Nosema ceranae and Nosema apis are microsporidia which play an important role in the epidemiology of honeybee microsporidiosis worldwide. Nosemiasis reduces honeybee population size and causes significant losses in honey production. To the best of our knowledge, limited information is available about the prevalence of nosemiasis in Italy. In this research, we determined the occurrence of Nosema infection in Central Italy. Thirty-eight seemingly healthy apiaries (2 to 4 hives each) were randomly selected and screened from April to September 2014 (n = 11) or from May to September 2015 (n = 27). The apiaries were located in six areas of Central Italy, including Lucca (n = 11), Massa Carrara (n = 9), Pisa (n = 9), Leghorn (n = 7), Florence (n = 1), and Prato (n = 1) provinces. Light microscopy was carried out according to current OIE recommendations to screen the presence of microsporidiosis in adult worker honeybees. Since the morphological characteristics of N. ceranae and N. apis spores are similar and can hardly be distinguished by optical microscopy, all samples were also screened by multiplex polymerase chain reaction (M-PCR) assay based on 16S rRNA-gene-targeted species-specific primers to differentiate N. ceranae from N. apis. Furthermore, PCR-positive samples were also sequenced to confirm the species of amplified Nosema DNA. Notably, Nosema spores were detected in samples from 24 out of 38 (63.2%, 95% CI: 47.8-78.5%) apiaries. Positivity rates in single provinces were 10/11, 8/9, 3/9, 1/7, or 1/1 (n = 2). A full agreement (Cohen's Kappa = 1) was assessed between microscopy and M-PCR. Based on M-PCR and DNA sequencing results, only N. ceranae was found. Overall, our results highlighted that N. ceranae infection occurs frequently in the cohort of honeybee populations that was examined despite the lack of clinical signs. These findings suggest that colony disease outbreaks might result from environmental factors that lead to higher susceptibility of honeybees to this microsporidian.

Molecular detection of tick-borne pathogens in wild red foxes (Vulpes vulpes) from Central Italy.

Spleen samples from 153 red foxes, shot during regular hunting season in the province of Pisa (Central Italy), were examined to detect DNA of Anaplasma phagocytophilum, Ehrlichia canis, Coxiella burnetii, Francisella tularensis, Hepatozoon canis and Babesia sp./Theileria sp. DNA of vector-borne pathogens was detected in 120 (78.43%; 95% CI: 71.06-84.66%) foxes. Specifically, 75 (49%; 95% CI: 40.86-57.22%) animals scored PCR-positive per H. canis, 68 (44.44%; 95% CI: 36.42-52.69%) for E. canis, 35 (22.88%; 95% CI: 16.48-30.35%) for piroplasms (Theileria annae), 3 (1.96%; 95% CI: 0.41-5.62%) for C. burnetii and 1 (0.65%; 95% CI: 0.02-3.59%) for A. phagocytophilum. No positive reaction was observed for F. tularensis. Fifty-six animals (36.6%; 95% CI: 28.97-44.76%) were positive for two or three pathogens. Red foxes result to be involved in the cycle of vector-borne pathogens that are associated to disease in dogs and humans.

Occurrence of Toxoplasma gondii in Carcasses of Pigs Reared in Intensive Systems in Northern Italy.

To evaluate the occurrence of Toxoplasma gondii and to genetically characterize its isolates in carcasses of industrial fattening pigs, blood, diaphragm, and heart samples were collected from 375 carcasses of pigs slaughtered to be processed for Parma ham production. Pigs had been bred on approved farms (n = 75) located in the so-called Food Valley in Italy. Sera were examined for immunoglobulin G antibodies to T. gondii by modified agglutination test (MAT). Both heart and diaphragm samples from seropositive carcasses were processed for the presence of T. gondii DNA (B1 locus) by real-time PCR and high resolution melting (HRM) assay. Anti-Toxoplasma antibodies were detected in 2.1% of pig carcasses, with titers from 1:10 to 1:320. T. gondii DNA was detected in all (eight) seropositive carcasses and in 11 (5 heart and 6 diaphragm samples) of 16 samples; that is, it was detected in heart tissue in two subjects, in diaphragm tissue in three subjects, and in both muscle tissues in three subjects. Toxoplasma genotypes were determined in seven of eight carcasses: type III was identified in four carcasses, type II in two, and both III and II in one carcass. The serological findings and the molecular detection of T. gondii strains suggest that cured meat products obtained from industrially bred pigs may be potential sources of toxoplasmosis for humans. Our results provide novel, important information regarding the seroprevalence and molecular prevalence of T. gondii in intensively reared pigs within this specific region of Italy, particularly because Parma ham from this region is known and consumed worldwide. On-farm preventive measures combined with slaughterhouse monitoring of carcasses of pigs bred for cured meat production should never be overlooked to prevent the introduction of T. gondii into the food chain and to ensure safety for consumers of these products.

First report of Tunisian coastal water contamination by protozoan parasites using mollusk bivalves as biological indicators.

In order to establish seawater contamination by emerging protozoan parasites, we used qPCR to molecularly characterize and evaluate the parasitic burden of Giardia duodenalis, Cryptosporidium spp., Toxoplasma gondii, and Cyclospora cayetanensis in 1255 wild bivalve mollusks collected along the Tunisian coasts. T. gondii, G. duodenalis and C. cayetanensis were detected in 6.9% (99% CI=1.6-12.2%) pools of Ruditapes decussatus. None of the samples were found positive to Cryptosporidium spp.; 6.6% pools of R. decussatus were positive for T. gondii Type I, 1.6% for G. duodenalis assemblage A, and 1.6% for the association T. gondii Type I/C. cayetanensis/G. duodenalis assemblage A. R. decussatus harbored up to 77500 oocysts/sample of T. gondii, up to 395 cysts/sample of G. duodenalis, and 526 oocysts/sample of C. cayetanensis. These results provide the first evidence that the Tunisian coasts are contaminated by zoonotic protozoan parasites that can constitute a direct or indirect risk for human health.

Traditional Mediterranean plants: characterization and use of an essential oils mixture to treat Malassezia otitis externa in atopic dogs.

Several plants extracts from Mediterranean countries are traditionally employed in skin troubles both in humans and in animals. Malassezia pachydermatis is a lipophylic yeast responsible for otitis externa and dermatitis in dogs and for cutaneous and systemic disease in humans. Five mixtures of essential oils obtained from Mediterranean plants (Citrus paradisi, Salvia sclarea, Ocimum basilicum, Rosmarinus officinalis, Citrus limon, Anthemis nobilis, Lavandula hybrida and Thymus vulgaris) provided with antifungal and/or anti-inflammatory action assayed in vitro, were tested in vivo versus M. pachydermatis to treat once daily for 2 weeks 25 atopic dogs with Malassezia otitis externa. Mixture composed by C. limon 1%, S. sclarea 0,5%, R. officinalis 1%, A. nobilis 0,5% yielded excellent results in all treated dogs. Despite of clinical resolution after all treatments the number of blastospores did not decrease. This study confirms recent findings suggesting a multifactorial alternative approach for the management of canine Malassezia otitis.

Open-field study comparing an essential oil-based shampoo with miconazole/chlorhexidine for haircoat disinfection in cats with spontaneous microsporiasis.

Objectives The goal of the present study was to compare the antifungal efficacy of an essential oil (EO) shampoo proven to be effective against Microsporum canis with miconazole/chlorhexidine for topical haircoat disinfection in cats treated concurrently with oral itraconazole. Methods Cats received treatment with oral itraconazole (Itrafungol) at a dose of 5 mg/kg/day pulse administration for 1 week, every 2 weeks for at least 6 weeks and were washed twice a week with a neutral shampoo with added EOs of Thymus serpyllum (2%), Origanum vulgare and Rosmarinus officinalis (5% each) for the period of systemic treatment. This protocol was compared with a conventional treatment (oral itraconazole + 2% miconazole/2% chlorhexidine shampoo). Results The treatment was well tolerated and adverse effects were not recorded. All cats were clinically negative at week 11. With respect to animals with extensive lesions, the speed of resolution was higher in cats with focal lesions. The animals showing diffuse lesions required more than a course of treatment to achieve a mycological cure. There was no significant difference between the number of weeks to obtain mycological cure for cats treated with EOs and animals treated conventionally. Conclusions and relevance The treatment appeared to be effective and well appreciated by the owners. The use of shampoo with the added EOs of T serpyllum, O vulgare and R officinalis would seem an interesting, natural alternative to conventional topical treatment.

Seroprevalence of Toxoplasma gondii and Neospora caninum in red deer from Central Italy.

Neospora caninum and Toxoplasma gondii are cosmopolite protozoan parasites impacting on human and animal health. In particular, T. gondii commonly infects human beings and all warm-blooded animals, while N. caninum is responsible for bovine abortion and neuromuscular disease in dogs. The aim of the presented survey was to evaluate the occurrence and prevalence of these parasites in the most numerous Italian red deer population. The sera of 60 red deer (Cervus elaphus) inhabiting Central Italy (43°56'N 10°55'E) and killed by selective hunting were examined using an indirect fluorescent antibody test (IFAT) for both N. caninum and T. gondii antibodies. White blood cells (buffy coat) were also checked by PCR and T. gondii DNA was genotyped. Thirteen out of 60 sera (22%) scored positive for Toxoplasma, 17 samples (28%) were Neospora positive. Coinfection was recorded in 5 cases (8%). T. gondii (genotype II) and N. caninum DNA was detected in one and 3 samples of buffy coat, respectively. The presented study is the first to examine the occurrence of these parasites in the most numerous red deer Italian population, confirming this animal species as carrier of the investigated pathogens. These animals spread near human settlements, co-inhabiting with final hosts of T. gondii and N. caninum and could contribute to their transmission to domestic ruminants and humans. In particular, the seroprevalence value for N. caninum was the highest among European records.