Anna Paola Pipia

Leishmaniasis is a widespread, vector-borne parasitosis causing clinical manifestations in animals and in humans. In dogs, Canine Leishmaniasis has been reported in as much as 50 countries and the Mediterranean basin is known to be one of the most affected zones. Within these areas, the Island of Sardinia (Italy) has long been considered endemic for leishmaniasis and the presence of two arthropod vectors has recently been reported there. Nevertheless, to date, no epidemiological surveys regarding CanL have been carried out on the island. Hence, for the first time, the seroprevalence and the risk factors were investigated. Blood samples, as well as clinical and general information from 1.147 dogs, were collected and analyzed. Dogs consisted of two distinct populations, namely “owned dogs” and “kennel dogs.” Anti-Leishmania IgG antibodies were detected using IFAT and samples were scored as positive at a cut-off dilution of 1:80. Data was analyzed using a Chi-squared test and bivariate and multivariate analyses were performed. Overall, 15.4% of dogs were found to be infected with CanL while only 44.1% of these animals exhibited clinical signs. Owned dogs (27.2%) were found to be infected more often than kennel dogs (10.6%); male dogs were found to be more frequently infected than female dogs and the number of infected animals increases with age. The present survey confirmed the endemic nature of leishmaniasis in Sardinia with a similar seroprevalence as mainland Italy. The results obtained serve as validation for the hypothesis that, in endemic areas, clinical CanL representations constitute only a fraction of the leishmaniasis cases.

This study is the first contribution to the molecular taxonomy of Mesocestoides spp. from domestic and wild carnivores in the Mediterranean area. A total of 13 adult worms and 13 larval stages of Mesocestoides spp. were collected from domestic and wild carnivore hosts in Italy and Tunisia. Samples collected in the Slovak Republic were used as comparative samples from outside the Mediterranean. The genes cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) of the mitochondrial genome were used as molecular markers to investigate the presence of cryptic Mesocestoides species in the area analysed. Results were consistent in showing three well-supported clusters of Mesocestoides spp. in southern Italy and Tunisia, which were strongly divergent from Mesocestoides litteratus, M. corti and M. lineatus. High levels of genetic variation and no evidence of geographical structuring was found between the clusters. Considering the low dispersal capability of the intermediate hosts of Mesocestoides spp., the lack of geographical structuring among the Mediterranean regions could be due to a high potential for dispersion of the definitive hosts. This study provides a foundation for future formal descriptions of new species of the genus Mesocestoides in the Mediterranean area.

Introduction: Cystic Echinococcosis (CE) is one of the most widespread zoonosis of veterinary and medical importance still constituting a sanitary, economic and socio-cultural problem in Italy. Methodology: The aim of this study was to update epidemiological data on cattle CE in Italy. Data on CE positivity of 5,336 cattle were acquired from abattoir registers between January 2009 and July 2010. Morphobiological characterization of hydatids was performed by direct examination of liver and lungs of 1,664 animals butchered in the same slaughterhouses in 2010. Strain typing of parasites was carried out through the amplification and sequencing of nd1 and cox1 mitochondrial genes. Results: Overall CE prevalence was of 8.1% (430/5,336). Parasitological examination of hydatids showed an overall prevalence of 8.6% with a fertility rate of 0.7% (12/1,664). Regarding localization, hydatids were found in 8% of the livers and in 7.6% of the lungs, respectively. Among positive animals, higher prevalence was observed in the liver (93%) compared to lungs (88.1%) (p > 0.05). Conclusion: The economic loss due to organs condemnation related to CE in cattle amounted to almost € 24,000 per year in the examined abattoir during 2010. Sequence analysis showed the presence of G1 (sheep strain) or Echinococcus granulosus sensu strictu in all examined samples. The G1 confirmed, once more, its possible development into several intermediate hosts such as cattle, especially in areas like southern Italy and Sardinia where the lifecycle of the parasite is still to date carried on by sheep and dogs.

Cystic echinococcosis (CE) is a zoonotic disease caused by Echinococcus granulosus. It is of worldwide importance, and is widespread in the Mediterranean region and Middle East. This tapeworm shows great intraspecific variation in relation to host specificity, epidemiology and morphology. This variability led in previous years to the identification of ten (G1-G10) different genotypes of the parasite. Cerebral localization of E. granulosus is not common: it especially affects children and is more frequently located in the supratentorial region. It can be life-threatening due to its localization in eloquent areas especially in the posterior fossa. Despite the benign nature of hydatid cyst, invasion of critical areas may cause significant mortality and morbidity in some patients. Urgent surgical decompression and adjuvant medical treatment must be employed as soon as possible in these patients. We present a clinical case of life-threatening brainstem compression in a child due to a rare form of CE which was confirmed with biomolecular techniques. She presented with respiratory distress and progressive quadriparesis. All cysts were removed by microsurgical technique and albendazole was given postoperatively for one year with regular follow-ups.

Shepherd and stray dogs are thought to represent the primary definitive hosts of Coenurosis by Taenia multiceps, due to their feeding habits which translate into high chances of coming into contact with infected intermediate hosts. Nonetheless, little attention has been paid to the role of the red fox (Vulpes vulpes) in the epidemiology of coenurosis. In fact a knowledge gap exists on the role played by red foxes in the epidemiology of Taenia multiceps and the capability of this parasite to produce fertile and viable eggs in this wild canid, i.e. on the occurrence of a sylvatic cycle. This study investigates the role of the red fox (Vulpes vulpes) in the epidemiology of T. multiceps and related metacestodoses. The small intestine of 63 red foxes was macroscopically examined for the presence of cestodes. Adult parasites were identified morphologically as being T. multiceps. Tapeworm eggs were counted and stored at 4 °C in physiological saline solution prior to experimental infection of four sheep and one goat. Sheep were inoculated orally on Day 0 with 3000 (sheep 1), 5000 (sheep 2 and 3) or 7000 eggs (sheep 4), while the goat was infected with 5000 eggs of T. multiceps. The animals were followed-up regularly by MRI and underwent surgical treatment between days 180 to day 240 post infection. Collected coenuri were identified using morphological and molecular methods. A total of 6.3 % of red foxes were found infected with T. multiceps and the eggs obtained from the worms were determined to have a viability of 45.4 %. Two of the challenged sheep and the goat developed disease compatible with T. multiceps. Morphometrical features of the cysts were consistent with those of T. multiceps; nucleotide amplification and sequencing of mitochondrial genes (i.e., cox1 and Nd1) from the metacestode material confirmed the identification. The present study is the first to provide evidence of the role of the red fox as a competent definitive host for T. multiceps, thus changing the epidemiological scenarios of infections by this cestode.

The aim of this study was to investigate the epidemiology and transmission of Taenia hydatigena in sheep and dogs from Sardinia and the economic estimation of losses due to this metacestodosis in lambs. A total of 7781 Sarda breed lambs were examined at abattoirs for the detection of Cysticercus tenuicollis or necrotic–haemorrhagic tracks of their migration. Morphological and molecular identification of parasites was carried out. Individual faecal samples from 300 dogs were examined for copromicroscopic investigations and coproELISA assay. An overall prevalence of 14.6 % for T. hydatigena cysticercosis was found in the examined lambs. In total, 10,807 parasitary tracks were found, with an abundance of 1.39 and an average intensity of 9.52. The molecular analysis of the isolates showed an overall pairwise nucleotide divergence for the CO1 and ND1 was of 0–3.1 and 0–3.3 %, respectively. Low intra- and interspecific variation was recorded for C. tenuicollis isolates used in this study which suggested the absence of differentiation. Microscopic examination of dog faeces showed a total prevalence of 31.3 % for endoparasites in the examined samples (94/300). Taeniid eggs were found in 8.3 % of the dogs. The results of the monoclonal antibody ATH4 ELISA test showed a prevalence of 11 % (33/300) for T. hydatigena coproantigens. The total economic costs related to cysticercosis amounted to almost € 330,000. The prevalence of C. tenuicollis in 14.6 % of 30–40-day-old lambs highlights the high parasitic pressure by T. hydatigena in the territory of Sardinia, Italy.

Natural infection with a species of Angiostrongylus has been reported only once in wildcats from central Italy by Biocca in 1957. The causative species of this infection was identified as Angiostrongylus chabaudi. Following this report, this parasite had never been found in either wild or domestic cats. The lungs and the pulmonary arteries of an adult female cat (Felis silvestris catus), road-killed in Sardinia, Italy, were macroscopically examined and dissected under a light microscope for the presence of parasites. A slender nematode was detected and its morphometrical features were consistent with those of A. chabaudi. Morphological data were supplemented by sequencing of the partial cytochrome oxidase c subunit 1 (cox 1) gene, as well as the internal transcribed spacer 2 (ITS2) of the rDNA. Nucleotide sequences displayed 99% homology with the ITS2 sequence [GenBank KM216825.1] of a specimen of Angiostrongylus sp. recovered recently from the pulmonary artery of a wildcat in Germany and 91% with cox 1 sequence [GenBank GU138118.1] of Angiostrongylus vasorum. The results of the present study indicate, for the first time, that A. chabaudi may also infect domestic cats, and thus should be considered in the diagnosis of metastrongyloid species infecting their cardio-pulmonary system.

Besides Aelurostrongylus abstrusus, other parasites belonging to the superfamily Metastrongyloidea, namely Oslerus rostratus, Troglostrongylus brevior and to the family Trichuridae, i.e. Eucoleus aerophilus (syn. Capillaria aerophila), have also been reported as agents of respiratory infection in domestic cats. A case of simultaneous infection by four feline lungworm species in Sardinia is herein described. An adult female cat (Felis silvestris catus), road-killed in the southeast part of Sardinia (municipality of Villacidro, province of Cagliari), Italy, was referred to the Laboratory of Parasitology of the Veterinary Teaching Hospital in Sassari. At necropsy, the lungs were examined and dissected under a stereomicroscope for the presence of parasites, and first-stage larvae (L1) of broncho-pulmonary nematodes were searched for in a faecal sample using the Baermann method. Parasites collected in the lungs were morphologically identified as A. abstrusus, E. aerophilus, and O. rostratus. In addition to the above species, L1s of Troglostrongylus spp. were detected at coproscopy but no adult specimen was found in the lungs. The morphological identification was confirmed by the molecular amplification and sequencing of cox1 mitochondrial gene, 18S and ITS2 ribosomal DNA. This finding stands as the first simultaneous infection by four feline lungworm species in the same animal, and as the first report of O. rostratus and E. aerophilus in Sardinia.