Gianluca Fichi

A comparison of coprological, serological and molecular methods for the diagnosis of horse infection with Anoplocephala perfoliata (Cestoda, Cyclophyllidea).

Anoplocephala perfoliata (Cestoda, Cyclophyllidea), the commonest intestinal tapeworm of horses, can cause colic, intussusceptions, ileal impactions and intestinal perforations. Common diagnostic techniques for A. perfoliata infection, i.e. coprology and serology, show inherent limitations in terms of sensitivity and specificity and new approaches are thus required. Hence, the present study compared the reliability of coprological, serological (i.e. ELISA) and molecular (i.e. nested PCR) methods in detecting A. perfoliata infection in naturally infected horses and in horses treated with a combination of ivermectin and praziquantel. Of 42 horses subjected to coprological examination, 16 and 26 resulted negative and positive, respectively for the presence of A. perfoliata eggs at the coprological examination. The 26 coprologically positive animals were also positive by nested PCR. Fifteen out of the 16 horses coprologically negative were negative at the molecular assay, while one yielded a PCR product detectable on an agarose gel. Eighteen out of 26 positive horses were treated with a combination of ivermectin 18.7 mg/g and praziquantel 140.3mg/g and resulted subsequently negative by coprology and nested PCR performed 2 weeks after treatment. All infected and untreated animals had a high ELISA test optical density indicating high infection intensity and associated risk of colic. However, high optical density values were also obtained in four horses post-treatment and in three horses that were negative on molecular and coprological analysis. The results of the present work indicate that the nested PCR assay represents a valid method for the specific molecular detection of A. perfoliata in faecal samples collected from naturally infected horses and may have advantages over coprological and serological approaches for diagnosing A. perfoliata infection.

Internal and External Parasitic Infections of Pet Reptiles in Italy

Abstract In captive reptiles, parasites may represent a serious health and management problem for both the reptiles and their human caretakers (zoonotic parasites). In the present study, the prevalence of ecto- and endoparasitic infections and the clinical signs associated with these parasites were determined in pet reptiles living in Italy. To accomplish these aims, fecal samples collected from 120 captive reptiles representing 27 different species, including 63 household pet and 57 pet store reptiles, were analyzed macroscopically and microscopically for the isolation and identification of endoparasites. In addition, the reptiles, along with their terrariums and water containers, were also examined for ectoparasitic arthropods. Recovered arthropods were preserved in 80% ethanol and evaluated microscopically. Overall, 72.5% of the examined reptiles were found to be positive for parasites, with prevalence rates of 45% and of 27.5% for endo- and ectoparasites, respectively. Pet shop animals were more likely to be infected with endoparasites, whereas ectoparasites were more prevalent in household animals. Isolated endoparasites included protozoans (Entamoeba sp., Balantidium sp., Isospora jaracimrmani, and Caryospora corallae) and nematodes (ascarids, pinworms, and strongylid nematodes), while ectoparasites included mites (Ophionyssus natricis, Hirstiella stamii, and Hirstiella diolii) and a species of tick (Amblyomma latum). This study is the first report of H. stamii, H. diolii, and A. latum in Italy. Results of this study confirm that pathogenic and potentially zoonotic parasites are common among pet reptiles in Italy and highlight the role of veterinarians in educating the public regarding the need to control pet reptile parasitic diseases and prevent the introduction of exotic reptile parasitic species.

In Vitro Anthelmintic Activity of Four Plant-Derived Compounds against Sheep Gastrointestinal Nematodes

By using the egg hatch test (EHT), the larval development test (LDT) and the larval mortality/paralysis test (LMT), the in vitro anthelmintic activity on sheep gastrointestinal strongyles (GIS) of four plant-derived pure compounds, mangiferin (at 0.25%, 0.125% and 0.0625%), rutin (at 1%, 0.75%, 0.5%), quercetin (at 1%), and β-sitosterol (at 1%, 0.75%, 0.5%), was investigated. For comparison, untreated and treated (0.1% thiabendazole, 0.1% TBZ) controls were used. Six repetitions were made throughout the experiment. Data were statistically elaborated using the χ2 test. The concentration able to inhibit the development of the 50% of L1s to L3s and causing the mortality of the 50% of L3s (EC50) was also calculated. L3s recovered from untreated Petri dishes were identified at the genus level. In EHT, all tested compounds at all concentrations significantly (p < 0.01) inhibited the hatch of the eggs when compared to the untreated controls, but none of them was as effective as 0.1% TBZ. In LDT, rutin (at 1%, 0.75% and 0.5%), mangiferin (at 0.25% and 0.125%), β-sitosterol (at 1%) and 0.1% TBZ completely prevented the larval development from L1 to L3 in respect to the untreated controls (p < 0.01). In LMT, all tested compounds significantly (p < 0.01) increased the death of L3s compared to the untreated controls, except for β-sitosterol at 0.5%. However, only rutin at all concentrations and 0.25% and 0.125% mangiferin were as effective as 0.1% TBZ. Haemonchus, Trichostrongylus, Chabertia and Teladorsagia/Ostertagia GIS genera, were identified.

Leishmania infantum, Dirofilaria spp. and other endoparasite infections in kennel dogs in central Italy

Prevalence and risk factors of Leishmania infantum, Dirofilaria spp. and other potentially zoonotic or canine-specific endoparasite infections were assessed in 639 kennel dogs from central Italy. To this end, individual blood and fecal samples were examined using parasitological, immunological and molecular techniques. The presence of compatible clinical pictures, as well as age and gender were considered as putative risks factors. To evaluate risk factors, multivariable analysis with logistic regression and univariable analysis with a Chi square test and a Fischer’s exact test were performed. Overall, 52.6% of dogs (95% CI 48.6-56.5) were found positive, while 39.6% of dogs (95% CI 35.8-43.5) were infected by potentially zoonotic species. Leishmania infantum and Dirofilaria repens showed prevalences of 2.5% (95% CI 1.5-4.1) and 2.8% (95% CI 1.7-4.5), respectively. The prevalence of cardiorespiratory parasites was 7.8% (95% CI 5.9-10.3) and included the species Angiostrongylus vasorum, Eucoleus aerophilus, Eucoleus boehmi and D. immitis; the latter showed a prevalence of 0.2% (95% CI 0.001-1). Intestinal parasites were significantly prevalent (38.8%, 95% CI 35-42.7) and they consisted mainly of species of major zoonotic concern, including ancylostomatids, Toxocara canis, Giardia duodenalis, Dipylidium caninum, Taeniidae, Strongyloides stercoralis and Cryptosporidium parvum. Endoparasites were significantly prevalent in clinically suspected dogs. Leishmania infantum and cardiorespiratory nematodes were prevalent in older dogs, while intestinal parasites were prevalent in younger dogs. Results show high dog and public health risks in kennels in central Italy, and suggest the need for more effective control measures.

Internal and external parasitic infections in captive psittacine birds

Parakeets and parrots can be infected by several parasitic species that may be either primary or opportunistic pathogens and may be responsible for mild or severe clinical forms of disease (Greiner and Ritchie 1994, Doneley 2009). Some psittacine parasitic species are host specific, while others can infect a wide range of animals—including humans (Tsai and others 1992, Fayer 2010). The main aim of this study was the evaluation of the prevalence of ectoparasites and endoparasites in captive psittacine birds living in Italy. In the period 2004–2007, 573 randomly selected captive psittacine birds of 47 different species living in Italy were examined. In this prospective survey all animals, including 450 farmed and 123 pet psittacines, were screened for endoparasites, while 196 farmed and 25 pet birds were examined also for ectoparasites. For endoparasites, fresh individual faecal samples were analysed by flotation test, a commercial immunoassay (RIDAQUICK Cryptosporidium / Giardia Combi cassettes, R-Biopharm, Italy) and by Giemsa and modified Ziehl Neelsen stained faecal smears. In addition, a deceased Platycercus elegans and a deceased Ara chloroptera were necropsied and examined for gross lesions and parasites. Isolated nematodes were identified at the species level (Levine 1968, Kajerova and others 2004a, b). Species of collected ectoparasites were also identified (Krantz 1978, Atyeo and Gaud 1987, Mironov and Galloway 2002, Schmaschke and others 2002, Sychra and others 2007, Di Palma and others …