Efficacy of a topical formulation containing eprinomectin, esafoxolaner and praziquantel (NexGard® Combo) in the treatment of natural respiratory capillariosis of cats.

Feline pulmonary capillariosis is a significant disorder due to its distribution and clinical impact. This study evaluated the safety and efficacy of two administrations 28 days apart of a topical solution containing esafoxolaner, eprinomectin and praziquantel (NexGard® Combo) in treating Eucoleus aerophilus (syn. Capillaria aerophila) infection in naturally infected cats. Cats were allocated to two groups: G1 cats (n = 23) received two treatments at study days (SDs) 0 and 28 (±2) and were evaluated for 6 weeks, and G2 cats (n = 17) served as a negative control for 6 weeks and were then treated twice on SDs 42 (±2) and 70 (±2), allowing for an additional 6-week assessment of efficacy. Each cat was subjected to McMaster coproscopy at SDs -7/0, 28 (±2) and 42 (±2) for both groups, 70 (±2) and 84 (±2) only for G2. Clinical examination and chest radiographic images were performed at SDs 0, 28 (±2) and 42 (±2) for G1 and G2, 70 (±2) and 84 (±2) only for G2. The comparison of EPG (eggs per gram of feces), clinical (CS), and radiographic scores (RS) at each time-point was used as a criterion. The efficacy based on the EPG reduction was 99.5% (G1) and 100% (G2) after two administrations of NexGard® Combo 2 weeks apart. At SD 0, no significant differences for CS and RS were recorded between G1 and G2, while a significant reduction (p < 0.05) was observed post-treatment for CS, RS, oculo-nasal discharge, auscultation noises, and cough. Two doses of NexGard® Combo 28 days apart stopped egg shedding and significantly improved clinical alterations in cats infected by E. aerophilus.

Title: Efficacité d'une formulation topique contenant de l'éprinomectine, de l'esafoxolaner et du praziquantel (NexGard® Combo) dans le traitement de la capillariose respiratoire naturelle du chat. Abstract: La capillariose pulmonaire féline est un trouble important, de par sa répartition et son impact clinique. Cette étude a évalué l'innocuité et l'efficacité de deux administrations à 28 jours d'intervalle d'une solution topique contenant de l'esafoxolaner, de l'éprinomectine et du praziquantel (NexGard® Combo) dans le traitement de l'infection à Eucoleus aerophilus (syn. Capillaria aerophila) chez des chats naturellement infectés. Les chats ont été répartis en deux groupes : les chats G1 (n = 23) ont reçu deux traitements aux jours d'étude (JE) 0 et 28 (±2) et ont été évalués pendant 6 semaines et les chats G2 (n = 17) ont servi de contrôle négatif pendant 6 semaines, puis ont été traités deux fois aux JE 42 (±2) et 70 (±2), permettant une évaluation supplémentaire de l'efficacité sur 6 semaines. Chaque chat a été soumis à une coproscopie McMaster aux JE −7/0, 28 (±2) et 42 (±2) pour les deux groupes, 70 (±2) et 84 (±2) uniquement pour G2. L'examen clinique et les images radiographiques thoraciques ont été réalisés aux JE 0, 28 (±2) et 42 (±2) pour G1 et G2, 70 (±2) et 84 (±2) uniquement pour G2. La comparaison des nombres d'œufs par gramme de matières fécales (OPG), score clinique (SC) et score radiographique (SR) à chaque point ont été utilisées comme critères d'efficacité. L'efficacité basée sur la réduction de OPG était de 99,5 % (G1) et de 100 % (G2) après deux administrations de NexGard Combo à deux semaines d'intervalle. À JE 0, aucune différence significative pour SC et SR n'a été enregistrée entre G1 et G2, tandis qu'une réduction significative (p < 0,05) a été observée après le traitement pour SC, SR, écoulements oculo-nasaux, bruits d'auscultation et toux. Deux doses de NexGard® Combo à 28 jours d'intervalle arrêtent l'excrétion des œufs et améliorent considérablement les altérations cliniques chez les chats infectés par E. aerophilus.

Detection of zoonotic enteropathogens in captive large felids in Italy.

AIMS: Within the One Health paradigm, infectious disease surveillance have been developed for domestic and wild animals, leaving the role of captive non-domestic populations, especially felids in zoos and circuses, less explored. This study addresses the proximity of these captive animals to urban areas, necessitating focused monitoring for potential zoonotic enteropathogens. The present work aimed to investigate the presence of such zoonotic enteropathogens in faecal samples from captive large felid populations. METHODS AND RESULTS: A total of 108 faecal samples were collected in three circuses, five zoos and one rescue centre across Italy. Salmonella spp. isolation, serotyping and antimicrobial susceptibility testing were conducted on all samples. Additionally, 60 samples were also examined for gastrointestinal parasites using standard coprological techniques. Giardia spp. detection employed direct immunofluorescent staining and specific PCR, while Toxoplasma gondii was detected using PCR targeting B1 gene. A total of 51 Salmonella enterica subsp. enterica were isolated, with predominant serovariants including Infantis (43.1%), Coeln (11.8%) and Newport (11.8%). The captive felids likely act as asymptomatic carriers of foodborne Salmonella, with notable resistance ampicillin and trimethoprim-sulfamethoxazole, no resistance to enrofloxacin was noted. Microscopic analysis revealed Toxascaris leonina eggs in 11 faecal samples (18.3%) and Giardia duodenalis cysts in one animal (1.7%). CONCLUSIONS: Captive animals in public settings may act as sources of Salmonella infection and enteroparasitosis for both occupational and general exposure. The study emphasizes the role of captive animals in antimicrobial resistance dynamics, highlighting the need for routine pathogen screening in the management practices of zoological structures.

Genetic variability of Dirofilaria repens isolates from humans and dogs in Italy.

Dirofilaria repens is a paradigmatic example of an emerging vector-borne pathogen (VBP) in both human and veterinary fields. The spatial expansion and the increasing zoonotic impact of this VBP can be related to several drivers including the genetic structure of parasite populations. Italy is one of the European countries traditionally endemic with the highest incidence of canine and human cases of subcutaneous dirofilariosis. The present study aimed to assess the genetic identity and variability of D. repens isolates of human and canine origin from areas of Central Italy, compared with those isolated from different areas of Europe by sequence analysis of mtDNA genes (i.e., 12 S rDNA and cox1). A total of twenty isolates of D. repens were obtained from biopsies of subcutaneous and ocular cases of dirofilariosis occurring in 10 dogs and 10 humans. The sequence analysis of 12 S rDNA showed that all the sequences obtained clustered as a monophyletic group with a strong nodal support, indicating that all sequence types represented D. repens. The cox1 and the 12 S sequence analysis did not show host-related polymorphisms between human and dog-derived specimens. The sequence analysis of cox1 was performed including 8 additional sequences previously obtained from human and canine isolates in the same areas. Out of the 28 sequences analyzed, 20 were grouped in a haplogroup comprising 15 haplotypes (i.e., DR1, DR2, DR4, DR5, DR7, DR8, DR10-DR18), 2 sequences matched to DR9, reported for the first time in Italy, and 6 showed peculiar polymorphisms that were not previously described. The results obtained have implications for a better understanding of the epidemiology and phylogeography of this emerging vector-borne zoonotic parasite.

Pets, Wildlife and Parasites.

In our dynamic world, borders of different sorts are being rapidly altered or even erased [...].

The seroprevalence of canine dirofilariosis in dogs in the eastern coastal areas of China.

Background and objectives: The present study was carried out to assess the presence of canine Dirofilaria immitis infection in pet dogs in China. Materials and methods: From October 2018 to November 2019, a total of 216 sera were collected from pet hospitals in Shandong, Jiangsu, Shanghai, Zhejiang, and Fujian regions of Eastern China. The sera were tested by using a commercial canine heartworm antibody ELISA test kit. Results: 70.8% of the pets had significant clinical symptoms resembled to heartworm infection; the overall dirofilariosis positivity found was 12.5% (27/216); Significant positive rates differences were observed between symptomatic and asymptomatic dogs (P < 0.05) (i.e. 15.7% and 4.7% respectively).The prevalence of infection in Shandong Province (15.5%) was the highest among the surveyed areas, but the difference among the geographic regions was not statistically significant (P > 0.05). Furthermore, the prevalence detected in summer (28.2%) was significantly higher than in other seasons (P < 0.05). In addition, no significant difference was observed between male and female sex (P > 0.05). Conclusions: Altogether, these results suggest that an epidemic of dirofilariosis exists in eastern coastal China, as such preventive measures should be taken to control the spread of dirofilariosis to reduce the risk of human and pet infection with heartworm.

Horse and donkey parasitology: differences and analogies for a correct diagnostic and management of major helminth infections.

In June 2022, at the XXXII Conference of the Italian Society of Parasitology, the parallels of the main endoparasitic infections of horses and donkeys were discussed. Although these 2 species are genetically different, they can be challenged by a similar range of parasites (i.e. small and large strongyles, and Parascaris spp.). Although equids can demonstrate some level of resilience to parasites, they have quite distinct helminth biodiversity, distribution and intensity among different geographical locations and breeds. Heavily infected donkeys may show fewer clinical signs than horses. Although parasite control is primarily provided to horses, we consider that there may be a risk of drug-resistance parasitic infection through passive infection in donkeys when sharing the same pasture areas. Knowing the possible lack of drug efficacy (<90 or 80%), it is advocated the use of selective treatment for both species based on fecal egg counts. Adult horses should receive treatment when the threshold exceeds 200­500 eggs per gram (EPG) of small strongyles. Moreover, considering that there are no precise indications in donkeys, a value >300 EPG may be a safe recommendation. We have highlighted the main points of the discussion including the dynamics of helminth infections between the 2 species.

Wild Mesocarnivores as Reservoirs of Endoparasites Causing Important Zoonoses and Emerging Bridging Infections across Europe.

Mesocarnivores are small- or mid-sized carnivore species that display a variety of ecologies and behaviours. In Europe, wild mesocarnivores are represented by the red fox (Vulpes vulpes), the golden jackal (Canis aureus), the European wildcat (Felis silvestris), the Mustelidae of the genera Meles, Martes, Mustela, Lutra, the invasive species of raccoon dog (Nyctereutes procyonoides), raccoons (Procyon lotor), and American mink (Neogale vison). These abundant animals thrive in various habitats and often develop their activity close to human settlements. Thus, they may play an important role in the introduction, maintenance, and transmission of major parasitic zoonoses and promote bridging infections with domestic animals. Against this background, this article reports and discusses some of the most important endoparasites of wild mesocarnivores living in Europe, on the basis of their actual role as reservoirs, spreaders, or sentinels. The data derived from epizootiological studies in different European countries, and the proven or speculated implications of the detected endoparasites in human and domestic animals' health, are discussed. Through older and recent literature review, the state-of-the-art knowledge on the occurrence and prevalence of the parasites under consideration is presented, showing further, warranted investigations and the need for surveillance and vigilance.

Parasitological Survey in European Brown Hare (Lepus europaeus Pallas, 1778) Breeding Facilities in Southern Italy.

Parasites are considered important regulating factors of hosts' population dynamics, not only in free-ranging wildlife, but also in captive bred animals. To date, only few studies have been carried out to assess the parasitic communities of the European brown hare in Southern Italy, and only one focused on animals in captivity. The aim of the present survey was to assess the composition of the endoparasite community in game hares bred for restocking purposes. For this purpose, 215 fecal pools collected in eight different breeding facilities were examined by qualitative and quantitative coprological techniques. Parasites characterized by a direct life cycle, including six species of coccidia from the genus Eimeria and the nematode Trichostrongylus retorataeformis, proved to be the most prevalent parasites. Further helminthic infestations by Passalurus ambiguous, Strongylosides papillosus, Cittotenia spp. and Dicrocoelium dendriticum were also detected, but with an overall prevalence lower than 20%. The present study contributes to increasing knowledge on the health status of a poorly investigated species, and is useful for optimizing breeding efforts in captivity.

The Dangerous Side of Being a Predator: Toxoplasma gondii and Neospora caninum in Birds of Prey.

Toxoplasma gondii and Neospora caninum are apicomplexan protozoa of major concern in livestock and T. gondii is also considered one of the major threats and a public health concern. These protozoa have a wide range of intermediate hosts, including birds. The present work aimed to assess the prevalence of these cyst-forming parasites in migratory and sedentary birds of prey. The skeletal muscle and myocardium of 159 birds of prey from Central Italy, belonging to 19 species and recovered across 6 Wildlife Recovery Centers/Care structures along the Italian migratory route, were collected specifically for molecular (PCR) and for histopathological analysis to detect T. gondii and N. caninum. For the molecular analysis, genomic DNA was extracted. The DNA was tested by sequence typing, targeting GRA6, 529 bp repeated element, B1, PK1, BTUB, SAG2, alt.SAG2, and APICO genes for T. gondii and to end-point PCR targeting NC5 gene for N. caninum. Thirty-seven out of the one hundred and fifty-nine analyzed samples tested positive for T. gondii with a prevalence of 23.27% and nine for N. caninum, with a prevalence of 5.66%. Thirty-two sequences were obtained from the thirty-seven isolates of T. gondii. Among these, 26 presented alleles compatible with type I strain in 1 or more loci, 4 with type II strain and 2 consisted of atypical strains. Toxoplasma gondii genetic variability in birds of prey confirms previous findings of wildlife as reservoirs of atypical strains. Results from the histology showed few protozoal tissue cysts in skeletal muscle (n. 4) and hearts (n. 2).

Detection of Leishmania spp. in Chronic Dermatitis: Retrospective Study in Exposed Horse Populations.

Leishmania infantum is a protozoan causing human zoonotic visceral leishmaniasis (ZVL) and visceral-cutaneous canine leishmaniosis (CanL) in the Mediterranean Basin. L. infantum is able to infect a large number of wild and domestic species, including cats, dogs, and horses. Since the 1990s, clinical cases of equine leishmaniasis (EL), typically characterized by cutaneous forms, have been increasingly diagnosed worldwide. The aim of the present study was to evaluate the presence of clinical forms of EL in CanL-endemic areas in Italy, where exposure of equine populations was ascertained from recent serological surveys. For this purpose, formalin-fixed and paraffin-embedded skin biopsies of 47 horses presenting chronic dermatitis compatible with EL were retrospectively selected for the study and subjected to conventional and q-PCR. A singular positivity for L. infantum was found; BLAST analysis of sequence amplicons revealed a 99-100% homology with L. infantum sequences. The histological examination revealed a nodular lymphoplasmacytic and histiocytic infiltrate; immunohistochemistry showed rare macrophages containing numerous positive amastigotes. The present retrospective study reports, for the first time, a case of a cutaneous lesion by L. infantum occurring in an Italian horse. Pathological and healthy skin samples should be investigated on a larger scale to provide information on the potential clinical impact of EL in the practice, and to define the role of horses in epidemiological ZVL and CanL scenarios.

Retrospective Longitudinal Survey on Canine Vector-Borne Pathogens: Trends and Challenges of 10 Years of Activities of a Veterinary Blood Bank.

Canine vector-borne pathogens (CVBPs) represent a challenge for veterinary transfusion medicine, since some can be transmitted by blood transfusion and are of zoonotic concern. Epidemiological data on CVBPs, obtained during 10 years of pre-donor screening (2012−2021) by a veterinary blood bank in central Italy, were used to conduct a retrospective epidemiological longitudinal survey. The results were obtained using the Immunofluorescent Antibody Test (IFAT) conducted on sera in order to assess IgG antibodies against Leishmania infantum, Ehrlichia canis, Anaplasma phagocythophilum, Babesia canis, and Rickettsia conorii; the modified Knott's test and an ELISA kit were used to detect Dirofilaria immitis and Dirofilaria repens. In total, 324 out of the 1260 canine blood donors (25.71%) tested seropositive for at least one pathogen. The highest overall positive rate was detected for L. infantum (12.22%), followed by E. canis (2.30%), A. phagocytophilum (1.19%), D. repens (0.95%), D. immitis (0.32%), and B. canis (0.16%). From 2012 to 2014, a prevalence of 20.12% was recorded for R. conorii. Mixed infections were recorded in 21 dogs. For all the CVBPs investigated, significant differences (p < 0.05) were not observed over the period studied. The results evidenced a non-negligible prevalence of CVBPs in canine donors, which were selected based on strict criteria concerning regular endo- and ectoparasite controls. The results confirmed that the blood bank could be a reliable local epidemiological observatory. The need for implemented screening is discussed.

Comparison of immunohistochemical and qPCR methods from granulomatous dermatitis lesions for detection of leishmania in dogs living in endemic areas: a preliminary study.

BACKGROUND: In canine leishmaniosis (CanL) endemic areas, pathologists often receive skin biopsies for testing with histopathologic findings suggestive-but not conclusive for a definitive diagnosis-of CanL lesions. I the absence of data on the infective status of animals, the diagnosis can therefore be challenging. The aim of this retrospective study was to evaluate the ability of immunohistochemistry (IHC) and quantitative PCR (qPCR) methods to detect Leishmania infection in skin biopsies with a histopathologic diagnosis of lymphoplasmacytic/histiocytic and/or granulomatous dermatitis and to correlate the pattern, depth and severity of the histopathologic lesions with the parasite load detected by qPCR and IHC. METHODS: Thirty formalin-fixed, paraffin-embedded skin samples were evaluated by hematoxylin-eosin (H&E) staining, IHC, conventional PCR (cPCR) and qPCR. The severity, pattern and depth of the dermal inflammation and parasite load were graded. RESULTS: Leishmania was detected by H&E staining in 8/30 sections (26.66%) and by IHC in 14/30 samples (46.66%). Parasite DNA was detected in 14/30 samples (46.66%) by cPCR and in 21/30 samples (70%) by qPCR, with an extremely variable parasite load (1.32-62.700 copies). The level of agreement was fair between H&E staining and cPCR (κ = 0.32), and moderate between H&E staining and IHC (κ = 0.58). The level of agreement between IHC and cPCR was good (κ = 0.65); between IHC and qPCR, moderate (κ = 0.41); and between cPCR and qPCR, fair (κ = 0.28). A significant association was found between the severity of dermal inflammation and the parasitic skin load by IHC, although with weak linear correlation. CONCLUSIONS: Our study underlines the difficulty of obtaining a definitive diagnosis of CanL cutaneous lesions, even with the most accurate diagnostic tests currently available. Based on our results, no single test is suitable on its own for the diagnosis of cutaneous lesions caused by Leishmania. However, in the presence of a moderate/severe lymphoplasmacytic/histiocytic and/or granulomatous dermatitis, we suggest performing IHC, as in our study this technique proved to be the method with the highest discriminatory power to estimate the role of the parasite in skin lesions. In mild lesions, IHC loses its discriminatory power and should be effectively combined with techniques such as qPCR.

Simultaneous Exposure to Angiostrongylus vasorum and Vector-Borne Pathogens in Dogs from Italy.

Several drivers have recently fostered the expansion of Angiostrongylus vasorum throughout Europe, where Vector-Borne Pathogens (VBPs) are also spreading. However, the level of simultaneous risk of infection is still unknown in canine populations. This study evaluated the simultaneous exposure to A. vasorum and major canine VBPs in dogs of Italy. Sera of 294 dogs were subjected to two ELISAs, detecting A. vasorum circulating antigens and antibodies against the parasite, and to the following assays: (i) SNAP® 4DX (IDEXX Laboratories Inc.) detecting Dirofilaria immitis antigens, and antibodies vs. Borrelia burgdorferi, Anaplasma spp. and Ehrlichia spp. and (ii) IFAT for the detection of antibodies vs. Leishmania infantum, Babesia canis and Rickettsia conorii. Twenty-two (7.5%, CI: 4.8-11.1%) and six (2%, CI: 0.7-4.4%) dogs scored positive for circulating A. vasorum antibodies and antigens, respectively. Seventeen dogs (5.8%, CI: 3.4-9.1%) were positive for A. vasorum antibodies + at least one VBP, three (1%, CI: 0.2-3%) for A. vasorum antigen + at least one VBP, while one dog (0.3%, CI: 0.01-1.88%) was positive for A. vasorum antigen + A. vasorum antibodies + B. canis antibodies. These results show that dogs living in different regions of Italy are at risk of simultaneous infections with both A. vasorum and VBPs. Despite the same scenario being likely in other countries of Europe, the current knowledge is scant. Therefore, further studies are warranted to amplify current epizootiological information and to understand whether control programs should be improved.

Morishitium polonicum as a Cause of Severe Respiratory Disease in Eurasian Blackbirds (Turdus merula) in Central Italy.

Two Eurasian Blackbirds (Turdus merula) from central Italy were found with severe cyclocoelid trematodosis associated with airsacculitis. The birds were submitted with severe respiratory distress; one died shortly after hospitalization, while the second bird was euthanized. At necropsy, a massive presence of cyclocoelid flukes was observed in the coelomic cavity and air sacs of both birds. The air sacs were diffusely opaque, thickened, and covered by scant fibrinous exudate mixed with numerous parasites. Histologically, the air sacs showed diffuse and severe oedema with fibrinous exudate. Diffuse mononucleated and heterophilic infiltration mixed with multiple granulomas contained degenerated trematodes. Morishitium polonicum was identified using morphologic keys and molecular analysis of extracted DNA. Infections caused by M. polonicum are poorly documented in blackbirds and the findings in these birds support the pathogenic role of this trematode as a potential cause of death in blackbirds in Italy. Extended epidemiologic surveys are required to properly assess the potential importance of M. polonicum as a life-threatening pathogen in Blackbird populations.

Exposure to Major Vector-Borne Diseases in Dogs Subjected to Different Preventative Regimens in Endemic Areas of Italy.

Vector-borne diseases (VBDs) are globally widespread arthropod-transmitted diseases with a significant impact on animal and human health. Many drivers have recently spurred the geographic spread of VBDs in dogs. This study has evaluated the exposure to most important VBDs in dogs under different preventative treatments in different regions of Italy, i.e., Veneto, Friuli Venezia-Giulia, Umbria, Giglio Island (Tuscany), Abruzzo and Latium. Serological analyses were performed to detect antibodies against Leishmania infantum, Babesia canis, Anaplasma phagocytophilum/Anaplasma platys, Ehrlichia canis/Ehrlichia ewingii, Borrelia burgdorferi, Rickettsia conorii and the circulating antigen of Dirofilaria immitis. Dogs were categorized according to the treatment schedule usually received, and the association between seropositivity and possible risk factors was statistically evaluated. Overall, 124/242 (51.2%) dogs tested positive for at least one pathogen, while 34 (14.0%) were exposed to two or more pathogens. The most detected seropositivity was against R. conorii, followed by Anaplasma spp., L. infantum, B. canis, and the other pathogens under study. Significant statistical associations were found according to geographical provenance, history of tick infestation, lifestyle and inadequate prophylactic treatments. Random/irregular treatments have been identified as a clear risk factor. These results show that adequate prophylactic treatment protocols are overlooked by dog owners, despite the availability of several effective products, with possible implications in veterinary medicine and on public health.

Efficacy of milbemycin oxime/afoxolaner chewable tablets (NEXGARD SPECTRA®) against Capillaria aerophila and Capillaria boehmi in naturally infected dogs.

BACKGROUND: Capillaria aerophila and Capillaria boehmi parasitize the respiratory system of wild and domestic carnivores. Capillaria aerophila inhabits the trachea and bronchi of dogs and cats, while C. boehmi affects the nasal cavities and sinuses of dogs. In dogs the infection may be subclinical or characterized by varying respiratory distress. METHODS: The present study evaluated the efficacy of an oral formulation containing milbemycin oxime and afoxolaner (NEXGARD SPECTRA®) in dogs naturally infected with C. aerophila and/or C. boehmi from three enzootic areas of Italy. Dogs were enrolled pending fecal examination and molecular confirmation of respiratory capillarioses. Dogs were allocated in two groups: Group 1 (G1, 25 dogs), treated with a negative control product with no anthelmintic activity (afoxolaner, NEXGARD®), and Group 2 (G2, 26 dogs), treated with NEXGARD SPECTRA®. At the day of treatment administration (Day 0), all dogs were clinically examined. Dogs were again subjected to clinical and fecal examinations at Days 28 (± 4) and 56 (± 2). The primary criterion for treatment efficacy was the reduction of fecal Capillaria egg counts in G2 compared with G1. The regression of/recovery from baseline clinical signs was considered as a further efficacy criterion. RESULTS: Percentage reduction of fecal Capillaria egg counts in the NEXGARD SPECTRA® group compared to the control group was > 97% on Day 28 and 100% on Day 56, respectively (p < 0.05 for both time points). Twelve of the 13 dogs in the NEXGARD SPECTRA® group with respiratory signs prior to treatment were free of clinical signs at the end of the study. Conversely, the six control group dogs with respiratory signs prior to treatment remained symptomatic. CONCLUSIONS: Results of the present study showed that NEXGARD SPECTRA® was safe and highly efficacious in the reduction of C. aerophila and C. boehmi eggs after one treatment with a complete reduction of the egg output after the second administration associated with a recovery from respiratory signs.

Three new species of Cytauxzoon in European wild felids.

Protists of the genus Cytauxzoon infect a wide variety of wild and domestic felids worldwide. While the American Cytauxzoon felis has been well described, data on the European isolates of Cytauxzoon are still scant. The aim of the current study was to determine the genetic diversity of European Cytauxzoon spp. in wild felids across Europe by analyzing one nuclear and two mitochondrial genes, along with representative complete mitochondrial genomes. Overall, 106 biological samples from wild felids (92 from Felis silvestris and 14 from Lynx lynx) from Germany, Romania, Czech Republic, and Luxembourg were collected and screened for the presence of Cytauxzoon spp. using nested PCR protocols, targeting the highly conserved 18S rDNA, mitochondrial cytochrome b (CytB) and cytochrome c oxidase subunit I (COI) genes. Furthermore, 18 previously confirmed wild felid biological samples from Europe, and comparative material from USA positive for C. felis, were included in the study. In 18S rDNA sequences analyses, Cytauxzoon spp. from felids formed two separate clades of New World and Old World isolates, with a low inner diversity of the European clade. In contrast to 18S rDNA, the phylogenetic analyses of CytB and COI genes affirmatively revealed three highly supported clades, resulting in three defined genotypes. Similar intra- and interspecific variability of CytB and COI genes was observed in the case of different Babesia spp. Considering geography, host species and analyses of three genes, we conclude that the three detected genotypes of Cytauxzoon in European wild felids represent three new species, which we herein describe.

A case of a dog refractory to different treatments for pulmonary capillariasis.

Pulmonary capillariasis is a parasitic disease caused by the nematode Eucoleus aerophilus which affects wild and domestic carnivores. Currently, there are no anthelmintics approved for use in the treatment of dogs infected with E. aerophilus. The use of several anthelmintics has been reported in a few case reports and field efficacy studies in cats; much less is known on the treatment of dogs infected with E. aerophilus. The paper describes a case of a 4-month-old, mixed breed intact male referred to the Veterinary Teaching Hospital (VTH) of the Department of Veterinary Medical Science of the University of Bologna for a routine vaccination and tested positive for E. aerophilus. The dog has not been responding to three different administered treatments, such as moxidectin, fenbendazole, and milbemycin oxime. Eighteen months after the first fecal examination, owner has brought in the dog for a routine visit; a coprological examination was requested and performed resulting negative for parasites. Veterinary practitioners, parasitologists, diagnostic laboratories, and dog owners need to be aware of the increased danger of possible treatment failure when attempting to control parasitic infections for which there are no approved anthelmintics with established efficacies available for use.

Detecting antibodies to Leishmania infantum in horses from areas with different epizooticity levels of canine leishmaniosis and a retrospective revision of Italian data.

BACKGROUND: Leishmania infantum is a vector-borne pathogen endemic in countries in the Mediterranean basin, including Italy. Dogs act as the primary reservoir for this parasite, but other animal species may also be infected. Low-to-moderate seroprevalence levels of infection have been reported in apparent healthy equine populations in southern Europe, reinforcing the importance of exploring those species, including horses, that act as a food source for vectors and may thus participate in the epizoological scenario of canine leishmaniosis (CanL) and zoonotic visceral leishmaniosis (ZVL). Since little is known regarding the exposure to L. infantum in horses in Italy, we assessed the seroprevalence in healthy equine populations from different CanL endemic areas. METHODS: The survey was conducted on 660 apparently healthy horses distributed throughout central and northern regions of Italy between 2016 and 2019. Blood samples were collected and the presence of anti-Leishmania antibodies (IgG) was investigated by the immunofluorescence antibody test. Information on the location and altitude of the stables, along with the horses' breed, age, sex, and reproductive status was obtained by filling in a questionnaire. This was then used for statistical analysis by generalized linear models to explore risk factors associated with seroreactivity to L. infantum. RESULTS: An average seroprevalence of 13.9% was detected for L. infantum in the equine populations investigated, with statistically significant associations between seroprevalence, geographical variables (northern vs central Italy, origin and altitude) and individual factors (i.e. age and breed morphotype). CONCLUSIONS: Our results highlight that horses are frequently exposed to L. infantum. Further prevalence surveys in horses, also using direct methods (e.g. PCR), are warranted to clarify the role of these hosts in the epidemiology of Leishmania in Italy.

Sarcocystis gigantea infection associated with granulomatous eosinophilic myositis in a horse.

The only Sarcocystis species currently known to inhabit the fibers of skeletal and cardiac muscles in horses are S. fayeri, S. bertrami, and S. asinus. We describe herein the invasion of myofibers in a horse by S. gigantea, a sheep-specific species with low virulence in the original host. A hunter gelding was referred to a veterinary surgeon in Newmarket (UK). The anamnestic data reported that the horse had an initial history of swelling of the right forelimb with fluid on the front of the carpus and edema spreading up the forearm. Subsequently, 2 firm lumps were found on the left pectoral muscle adjacent to the axilla of the left forelimb. Histologic examination of biopsies from the lumps revealed multifocal granulomatous eosinophilic myositis associated with intact and degenerate encysted parasites, consistent with Sarcocystis spp. Based on amplification and DNA sequencing of the 18S rRNA gene obtained from formalin-fixed, paraffin-embedded tissue blocks, S. gigantea was identified. The presence of sarcocysts in equine skeletal muscles has been considered an incidental finding, and there are only sporadic associated reports of myositis. Our finding suggests that some Sarcocystis spp. have a wider intermediate host range than believed previously, and that Sarcocystis of other species (not considered horse-associated) can invade the muscle fibers of equids, leading to myositis.