Seropositivity for Toxoplasma gondii in cats with clinical signs and living in households with women of childbearing age.

Toxoplasmosis caused by Toxoplasma gondii is a protozoal zoonosis with high sanitary risk for pregnant women and immunocompromised people. Felids, including domestic cats, are the only definitive hosts of T. gondii. They shed oocysts which, in the environment, become infectious for a wide range of animals, including humans, acting as intermediate hosts. This study evaluated the frequency of acute toxoplasmosis in domestic cats with compatible clinical signs and living in households with women of childbearing age. Individual serum samples were collected from 150 cats and analyzed for IgM and IgG against T. gondii. Statistical analyses were performed to evaluate associations between seropositivity and potential risk factors. Overall, 31 cats (20.7 %) were seropositive for anti-T. gondii antibodies, i.e. 9 (6.0 %) for IgM, 17 (11.3 %) for IgG and 5 (3.3 %) for both. The cats showed different combinations of clinical pictures. The following statistically significant associations were found: male sex and positivity for IgM and/or IgG (p=0.0248; OR= 0.3537; 95 % CI= 0.1528-0.8675), presence of 2 or more clinical signs and positivity to IgM only (p=0.0003; OR= +infinity; 95 % CI= 3.924 to +infinity), presence of either neurological signs (p=0.0025; OR= 13.30; 95 % CI= 3.409-61.39) or ocular signs (p=0.0228; OR= 5.835; 95 % CI= 1.631-22.37) and positivity to IgM only, presence of gastrointestinal signs and positivity to IgG only (p=0.0083; OR= 5.508; 95 % CI= 1.503-18.54). Male sex also resulted a possible risk factor in the binomial logistic regression (p= 0.011; OR= 3.336; 95 % CI= 1.131-8.44). These results indicate that cats living with women of childbearing age are at risk of infection with T. gondii. The presence of certain clinical signs can be helpful in identifying recent and/or current infections using laboratory analyses. Awareness on toxoplasmosis should be kept high to protect animal and public health.

Clinical efficacy of a parasiticide formulation containing eprinomectin, esafoxolaner and praziquantel (NexGard® Combo) in the treatment of natural feline aelurostrongylosis and troglostrongylosis.

The metastrongyloids Aelurostrongylus abstrusus and Troglostrongylus brevior are primary causes of feline clinical respiratory disease. The present field trial evaluated the clinical efficacy of a spot-on formulation containing eprinomectin, esafoxolaner and praziquantel (NexGard® Combo) administered per label recommendations to cats affected with aelurostrongylosis and/or troglostrongylosis. Overall, 36 naturally infected cats were randomly assigned to Group 1 (G1) or Group 2 (G2) of 18 cats each. The two groups included 6 cats with A. abstrusus, T. brevior and mixed infection, each. All cats completed the study. Cats in G1 were treated on study Days (SDs) 0 and 28±2. Cats in G2 served as negative control until SD 56±2 and were then treated on SD 56±2 and 84±2. On SD 0/-7, 28±2 and SD 56±2 all cats were subjected to parasitological (quali-quantitative Baermann) and clinical examinations (physical exams and thoracic X-rays). Hematology and biochemistry analyses were performed on SD 0/-7 and SD 56±2. On SD 84±2 quali-quantitative Baermann, clinical examination and thorax radiography were performed on all G2 cats and on two G1 cats that still had radiographic alterations on SD 56±2. On SD 112±2 all G2 cats were subjected to parasitological and clinical evaluations and one cat from G1 that still had radiographic signs at SD 84±2 was clinically and radiographically evaluated. Efficacy criteria were the reduction of larval shedding in faeces and the clinical response in terms of pathological and radiographic scores after treatment compared to the baseline. An efficacy of 100 % based on LPG reduction was recorded after one (20/24 cats) or two (all 24 cats) treatments in cats with single infection by A. abstrusus or T. brevior. For cats with mixed infections, larval shedding was stopped after one (11/12 cats) or two (all 12 cats) treatments. Statistically significant clinical and radiographic improvement was evident in all study cats after 2 treatments. The present data show that two monthly treatments with NexGard® Combo stopped larval shedding and led to a significant clinical recovery and a complete resolution of radiographic abnormalities in cats infected with A. abstrusus and/or T. brevior.

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.

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.

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.

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.

Pilot study evaluating the efficacy of a topical formulation containing emodepside and praziquantel in the treatment of natural feline troglostrongylosis.

BACKGROUND: Troglostrongylus brevior, a lungworm usually affecting wild felids, has been recently recorded in a number of cases in domestic cats, mainly in Mediterranean areas. Although feline troglostrongylosis is a severe and life-threatening disease, especially in young cats, treatment options are very limited. The present study evaluated the efficacy and safety of a spot-on formulation containing emodepside 2.1% and praziquantel 8.6% (Profender®, Bayer), which is licensed for treatment of the more common cat lungworm Aelurostrongylus abstrusus, for the treatment of natural troglostrongylosis. METHODS: Sixteen cats enrolled in the study were 1:1 allocated to two groups, i.e. Group T, treated with Profender® spot-on on days 0 and 14 (± 2) at the recommended clinical dose, and Group C which remained untreated. After study completion, the control cats received two rescue treatments with Profender® on days 28 (± 2) and 42 (± 2). The primary efficacy criterion was the absence of T. brevior L1 following treatment. Other efficacy parameters were the quantitative comparison of L1 presence before (baseline) and after treatment in both groups, and the comparison of clinical signs pre- and post-treatment. RESULTS: In terms of stopping larval shedding, Profender® showed an efficacy of 97% and 97.5% (arithmetic and geometric means, respectively) for group T, 97.1% and 98.5% for group C after one administration, and 100% for both groups after two doses. Overall, 12 cats showed clinical signs related to T. brevior. Specifically, 9 were clinically affected before treatment while clinical signs appeared after the first treatment in 3 cats. At the end of the study, all symptomatic cats fully recovered with the exception of 3 cats that showed clinical signs similar to those observed at the pre-treatment examination at the end of the study. CONCLUSIONS: This study shows that Profender® is effective against T. brevior.

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.

Cyathostomin faecal egg counts in horse farms from Central Italy.

Cyathostomins,  or  'small  strongyles',  are  the  most  important  equine  helminths  because  of their  worldwide  distribution,  spread  of  anthelmintic­resistant  populations,  and  pathogenic impact.  The  so­called  'selective  treatment'  of  those  animals  exceeding  a  certain  faecal  egg  count  (FEC)  has  recently  been  proposed  to  implement  cyathostomin  control  programmes.  The present study evaluated the extent of egg shedding in 475 horses living in 12 farms from 3 regions of Italy. All examined farms and 224 horses (47.6%) were positive for cyathostomins. 138 horses (28.8%) scored positive for cyathostomin FECs with a range of eggs­per­gram of faeces (epg) values of 50­2,150. Further 86 horses (18.1%) were positive only under qualitative microscopy (i.e. < 50 epg). Of the animals with a FEC > 50 epg, 81 (17%) and 57 (12%) showed values  of  50­200  and  >  200  epg,  respectively.  The  findings  from  this  study  demonstrated  a reduced presence of high­shedding horses compared to results obtained in previous years in  the  same  study  areas,  despite  overlapping  climate  features  in  the  previous  5  years.  A  continuing  monitoring  of  epidemiological  and  biological  features  of  horse  cyathostomin infection  is  crucial  for  planning  intervention  programmes  aimed  to  maintaining  animal health and preserving the efficacy of parasiticides.

Helminth infections in faecal samples of Apennine wolf (Canis lupus italicus) and Marsican brown bear (Ursus arctos marsicanus) in two protected national parks of central Italy

This article reports the results of a copromicroscopic and molecular investigation carried out on faecal samples of wolves (n=37) and brown bears (n=80) collected in two protected national parks of central Italy (Abruzzo Region). Twenty-three (62.2%) samples from wolves were positive for parasite eggs. Eight (34.78%) samples scored positive for single infections, i.e. E. aerophilus (21.74%), Ancylostoma/Uncinaria (4.34%), Trichuris vulpis (4.34%), T. canis (4.34%). Polyspecific infections were found in 15 samples (65.21%), these being the most frequent association: E. aerophilus and Ancylostoma/Uncinaria. Thirty-seven (46.25%) out of the 80 faecal samples from bears were positive for parasite eggs. Fourteen (37.83%) samples were positive for B. transfuga, and six (16.21%) of them also contained Ancylostoma/Uncinaria, one (2.7%) E. aerophilus and one (2.7%) both E. aerophilus and Ancylostoma/Uncinaria. Of the other samples, 19 (51.35%) were positive for Ancylostoma/Uncinaria, two (5.4%) for E. aerophilus and two (5.4%) for both. Molecular analysis found the roundworm and capillariid eggs found in wolves and bear samples to be Toxocara canis, Baylisascaris transfuga and Eucoleus aerophilus (syn. Capillaria aerophila). Considering the high prevalence of zoonotic intestinal helminths detected in this study, it is important to improve the knowledge and awareness of the general public and park operators regarding the potential health risk associated with infections in wildlife.

Intestinal parasites and vector-borne pathogens in stray and free-roaming cats living in continental and insular Greece.

This survey investigated the distribution of various intestinal parasites and vector-borne pathogens in stray and free-roaming cats living in four regions of Greece. A total number of one hundred and fifty cats living in three Islands (Crete, Mykonos and Skopelos) and in Athens municipality was established as a realistic aim to be accomplished in the study areas. All cats were examined with different microscopic, serological and molecular assays aiming at evaluating the occurrence of intestinal parasites, and exposure to or presence of vector-borne infections. A total of 135 cats (90%) was positive for one or more parasites and/or pathogens transmitted by ectoparasites. Forty-four (29.3%) cats were positive for one single infection, while 91 (60.7%) for more than one pathogen. A high number of (n. 53) multiple infections caused by feline intestinal and vector-borne agents including at least one zoonotic pathogen was detected. Among them, the most frequently recorded helminths were roundworms (Toxocara cati, 24%) and Dipylidium caninum (2%), while a high number of examined animals (58.8%) had seroreaction for Bartonella spp., followed by Rickettsia spp. (43.2%) and Leishmania infantum (6.1%). DNA-based assays revealed the zoonotic arthropod-borne organisms Bartonella henselae, Bartonella clarridgeiae, Rickettsia spp., and L. infantum. These results show that free-ranging cats living in areas of Greece under examination may be exposed to a plethora of internal parasites and vector-borne pathogens, some of them potentially able to infect humans. Therefore, epidemiological vigilance and appropriate control measures are crucial for the prevention and control of these infections and to minimize the risk of infection for people.

Mitochondrial haplotypes of Aelurostrongylus abstrusus and Troglostrongylus brevior (Nematoda, Metastrongyloidea) from domestic and wild felids.

The lungworm Aelurostrongylus abstrusus is the most important respiratory parasite of domestic cats. Pulmonary aelurostrongylosis has been reported in wild felids, though unequivocally evidence of wildlife infection by A. abstrusus is scant. Recently, Troglostrongylus brevior, a lungworm usually infecting wild felids, has been described in domestic cats from Mediterranean areas. The present work evaluates the sequence variation of an informative region within the gene encoding the mitochondrial cytochrome c oxidase subunit 1 of A. abstrusus and T. brevior, in order to provide novel information on the genetic make-up of these lungworms. Parasitic stages of A. abstrusus and T. brevior were collected from domestic and wild hosts (i.e., domestic cat, European wildcat, caracal, serval, and lion) from Italy, Greece, and South Africa. Five (HI-HV) and four (HI-HIV) haplotypes were recorded for A. abstrusus and T. brevior, respectively, mostly shared between domestic and wild felids in different geographical areas. The phylogenetic analysis showed that all haplotypes of A. abstrusus and T. brevior clustered as monophyletic groups with a strong nodal support, indicating that all haplotypes identified were distinct from each other. All sequence types represent two distinct species, A. abstrusus and T. brevior, and these genetic convergences are also detected within and among populations of these nematodes, irrespective of their hosts and geographical origin. The occurrence of A. abstrusus and T. brevior haplotypes in different hosts from the same regions and between different countries indicates that the same lungworm populations circulate in domestic and wild hosts under the same routes of transmission.

Aelurostrongylus abstrusus in wild felids of South Africa.

The increasing interest on respiratory nematodes of domestic cats (Felis silvestris catus) has recently stimulated several studies on their epidemiology and geographic distribution. At the same time, there are still important gaps in our knowledge of the infections caused by respiratory nematodes in wild felids. The present study investigated the occurrence of pulmonary parasites in wild hosts housed in sanctuaries and protected areas of South Africa. Faecal samples collected from seven species of wild felids living in three study sites were copromicroscopically and genetically examined. Of twenty-one samples six, i.e. three from caracals (Caracal caracal), two from lions (Panthera leo) and one from a serval (Leptailurus serval), scored positive for the metastrongyloid Aelurostrongylus abstrusus at copromicroscopic and/or molecular tests. No other lungworms were recorded. The occurrence of the cosmopolitan cat lungworm A. abstrusus in wild hosts has been so far questioned. Importantly, the present findings represent an unequivocal evidence of the capability of A. abstrusus to infect some species of wild felids. Further studies are warranted to understand the epidemiological patterns of lungworms in wild and domestic felids, and to better investigate the impact of these parasitoses on health and welfare of wild animals.

Survey on intestinal helminth fauna of woodcocks (Scolopax rusticola) in Italy.

Every year populations of the European woodcock (Scolopax rusticola) migrate from Central and Northern Europe to the Mediterranean basin. South of Italy is one of the most common wintering site for this species. Given that information on parasites of woodcocks is scarce, the present study aimed at identifying the parasitic species affecting woodcocks migrating in Italy. The gastrointestinal tract of 206 woodcocks hunted in Southern Italy was removed and examined for parasites. From each animal a faecal sample was analysed by flotation test. The necropsy showed the presence of cestodes, i.e. Paricterotaenia paradoxa (59.4%) and Aploparaksis filum (49.5%), and of acanthocephalan Prosthorhynchus scolopacidis (22.4%). In one bird we also detected Parastrigea robusta, which is a trematode until now reported only in mallards (Anas platyrhynchos). Mixed infections (i.e., polyspecific infections) were detected in 53 (27.6%) animals. The most common were those caused by A. filum and P. paradoxa (12.5%), and by P. paradoxa and P. scolopacidis (8.3%). Copromicroscopic examinations revealed the presence of eggs belonging to nematodes Syngamus spp. (1.94%) in 4 woodcocks and of eggs of cestodes Aploparaksis spp. (37.86%) in 78 woodcocks. The present results fill a gap in the knowledge on parasites affecting woodcocks.

OCCURRENCE OF LUNGWORMS IN EUROPEAN WILDCATS (FELIS SILVESTRIS SILVESTRIS) OF CENTRAL ITALY.

The increasing focus on infections in domestic cats ( Felis catus ) has raised questions about lungworm distribution in wild hosts. To enhance knowledge of the occurrence of lungworms in enzootic regions of central Italy, we examined the carcasses of 16 European wildcats ( Felis silvestris silvestris). Adult nematodes, feces, respiratory flushings, and pulmonary tissues were collected at necropsy and then microscopically and genetically analyzed. Fourteen wildcats had single or mixed lungworm species. Aelurostrongylus abstrusus was the most common parasite retrieved, followed by Troglostrongylus brevior. In addition, three specimens of Angiostrongylus chabaudi were found in the pulmonary arteries of one wildcat. Histologically, the most common lesions were a mild-to-severe chronic catarrhal bronchitis and a chronic interstitial pneumonia with smooth muscle hypertrophy, associated with T. brevior and A. abstrusus , respectively. These results demonstrate that the European wildcats may harbor several species of lungworms that may impair their health and welfare. Also, F. s. silvestris is a potential reservoir for respiratory nematodes in domestic cats.

Neospora caninum seropositivity and reproductive risk factors in dogs.

Despite the importance of Neospora caninum in veterinary medicine, knowledge of distribution of neosporosis in dog populations in some countries is still poor. The aims of the present study were to determine the occurrence of anti-N. caninum antibodies in one-hundred dogs living in cattle farms or dog breedings in central Italy and to evaluate the risk factors associated with seropositivity. The incidence of reproductive system disorders (e.g. infertility after first pregnancy) was also evaluated. Serum from breeding and farm dogs was tested to an indirect immunofluorescent antibody test (IFAT) to assess the occurrence of seropositivity. Management and individual data were collected and analysed both by linear and logistic multiple-regression models to find reliable predictors of seroprevalence and anti-N. caninum antibody level. The seropositivity for N. caninum was 32%. Dogs reared for breeding and presence of cattle on the farm were associated with seropositivity for N. caninum. Dogs living in the cattle farms showed a higher seropositivity for N. caninum (46%) compared with those living in dogs breeding (18%) (P < 0.05). The high presence of seropositive dogs in cattle farms of the study region demonstrates the potential risk of horizontal transmission of N. caninum between dogs and cattle, regardless the occurrence of reproductive system disorders or with infectious bovine tissues contact. Although the Neospora seropositivity in dog breedings may appear relatively low if compared with that found in dogs living with livestock, this infection, apparently underestimated, should be considered as a potential serious problem in canine medicine.

Larval development of Angiostrongylus vasorum in the land snail Helix aspersa.

The metastrongyloid nematode Angiostrongylus vasorum affects the heart and pulmonary arteries of dogs and wild animals. Over the recent years, dog angiostrongylosis has gained great attention in the veterinary community for the expansion of its geographic range and for a rise in the number of clinical cases. Global warming, changes in phenology of mollusc intermediate hosts and movements of wild reservoirs have been evocated in the spreading of mollusc-borne parasites, including A. vasorum. The land snail Helix aspersa, a vector of other respiratory metastrongyloids, is endemic in most regions of the World, where it is a pest outside its native Mediterranean range. In the present study, the susceptibility and suitability of H. aspersa as an intermediate host of A. vasorum were investigated along with the characteristics of larval recovery and development following two different ways of inoculation, i.e. experimental (group A) vs natural infection (group B). After infections, the snails were kept at environmental conditions for 2 months. Five snails from groups A and B were randomly selected, digested and examined at 15-day intervals for 2 months. L1s, L2s and L3s were microscopically identified based on key morphological and morphometric characteristics and their identity was genetically confirmed. The results showed that A. vasorum may reach the infective stage in H. aspersa and that uptake of larvae and parasitic burden within the snails depend on the grazing capability of the molluscs. Biological and epidemiological implications are discussed.

Novel Molecular Assay for Simultaneous Identification of Neglected Lungworms and Heartworms Affecting Cats.

Feline lungworms and heartworms are stimulating the interest of the scientific community due to their clinical impact and apparent geographical expansion. Diagnosis of the infections caused by these nematodes is indeed challenging. This report describes a novel multiplex PCR able to identify simultaneously three species of lungworms (Aelurostrongylus abstrusus and Troglostrongylus brevior) and heartworms (Angiostrongylus chabaudi) affecting felids. Epidemiological and clinical perspectives are discussed.

PCR-based assay for the mitochondrial cox1 specific amplification of Eucoleus böhmi.

Eucoleus böhmi (syn. Capillaria boehmi) is a trichuroid nematode affecting the epithelium of the nasal turbinates, frontal and paranasal sinuses of wild and domestic canids. Knowledge of the geographic distribution of nasal eucoleosis is fragmentary, despite the infection has been described from Europe and North America. Moreover, gaps exist in information available on the importance of the disease in canine clinical practice. The lack of knowledge on E. böhmi is likely due to limitations inherent to diagnostic methodologies. The aim of the present work was to assess a PCR-based assay instrumental to the amplification of a species-specific region of the mitochondrial DNA (mtDNA) gene encoding for the subunit 1 (cox1) gene of E. böhmi. Adult worms of E. böhmi from red foxes and dogs from Norway, Serbia and Italy and individual fecal samples from naturally infected dogs from Italy were included in the study. Stool samples from dogs negative for E. böhmi, but positive for other common parasites in both single and mixed infections, and adult stages of common dog parasites, were used to assess the specificity of this genetic assay. Using the panel of faecal samples, the assay showed a sensitivity of 85.14% and a specificity of 100%.