Active and passive surveillance for bat lyssaviruses in Italy revealed serological evidence for their circulation in three bat species.

The wide geographical distribution and genetic diversity of bat-associated lyssaviruses (LYSVs) across Europe suggest that similar viruses may also be harboured in Italian insectivorous bats. Indeed, bats were first included within the passive national surveillance programme for rabies in wildlife in the 1980s, while active surveillance has been performed since 2008. The active surveillance strategies implemented allowed us to detect neutralizing antibodies directed towards European bat 1 lyssavirus in six out of the nine maternity colonies object of the study across the whole country. Seropositive bats were Myotis myotis, M. blythii and Tadarida teniotis. On the contrary, the virus was neither detected through passive nor active surveillance, suggesting that fatal neurological infection is rare also in seropositive colonies. Although the number of tested samples has steadily increased in recent years, submission turned out to be rather sporadic and did not include carcasses from bat species that account for the majority of LYSVs cases in Europe, such as Eptesicus serotinus, M. daubentonii, M. dasycneme and M. nattereri. A closer collaboration with bat handlers is therefore mandatory to improve passive surveillance and decrypt the significance of serological data obtained up to now.

Molecular and phylogenetic analysis of the filarial nematode Micipsella numidica from the hare Lepus europaeus in Italy.

The genus Micipsella comprises three species of filariae to date identified in lagomorphs only, whereas the other genera belonging to the subfamily Splendidofilariinae are described as parasites of birds, reptiles and mammals. In the present study seven specimens of Micipsella numidica (Seurat, 1917), collected from the hare Lepus europaeus in Italy, were characterized genetically by molecular amplification of the mitochondrial genes (12S rDNA; cox1) and the 5S rDNA gene spacer region. Phylogenetic trees inferred using available sequences from filariae and those identified in this study evidenced a close relationship between M. numidica and Splendidofilariinae of other mammals and reptiles (Rumenfilaria andersoni and Madathamugadia hiepei). The present findings, apart from adding new data about the hosts in Italy, support the taxonomic position of M. numidica and highlight the substantial biological and molecular differences existing between Splendidofilariinae and other Onchocercidae. The study also contributes to our knowledge of the molecular/genetic diagnosis of filarial parasites of veterinary and medical concern in any vertebrate or invertebrate host.

The European wildcats (Felis silvestris silvestris) as reservoir hosts of Troglostrongylus brevior (Strongylida: Crenosomatidae) lungworms.

The increasing reports of Troglostrongylus brevior lungworm in domestic cats from Italy and Spain raised questions on its factual distribution and on the role wildcats play as reservoirs of these parasites. Carcasses of 21 wildcats were collected in natural parks of southern Italy (i.e., Catania, Sicily n=5 and Matera, Basilicata n=16) and biometrically and genetically identified as Felis silvestris silvestris, but two as hybrids. Troglostrongylus brevior and Eucoleus aerophilus lungworms were found in 15 (71.4%) and 7 (33.3%) individuals, respectively, being five (23.8%) co-infected by the two species. Both lungworms showed an aggregated distribution in the host population, assessed by k-index (i.e., 0.69 for T. brevior and 0.42 for E. aerophilus). Although no statistical significant difference was recorded among age, gender and geographical location of wildcats, a larger rate of infection by T. brevior was assessed in yearlings (85.7%) than adults (64.3%). This is the first epidemiological study reporting T. brevior infection in the European wildcat and discusses the potential threat this may represent for the conservation of this endangered species of felids. In addition, given the large frequency of lungworm infection herein recorded, the role of wildcats as reservoir hosts of these parasites to domestic cats is discussed.

Cryptosporidium in pet snakes from Italy: molecular characterization and zoonotic implications.

To provide information on the occurrence of Cryptosporidium species and genotypes in captive snakes from Italy, faecal specimens from 120 snakes belonging to 13 different genera of the families Boidae, Colubridae and Pythonidae were collected. Faecal samples were taken from the ground of the terrarium when available; otherwise cloacal cotton swabs were used. No clinical signs of cryptosporidiosis were observed in any animal at the time of sampling. Samples were examined for the presence of Cryptosporidium by using a direct immunofluorescence antibody test (IFAT) and two-step nested PCR at the small subunit (SSU) rRNA locus. PCR-positive samples were genotyped by restriction fragment length polymorphism (RFLP) analysis with the endonucleases SspI and VspI. By IFAT, 42 out of 120 snakes (35.0%) were found to be shedding Cryptosporidium oocysts. A significant higher percentage of positive ophidians were detected by using faecal specimens obtained from the terrarium (55.5%) than by cloacal cotton swabs (29.0%). SSU rRNA gene products were obtained from 25 isolates. Twenty samples tested positive to both microscopy and molecular techniques. Our data reveal a wide extent of cryptosporidial infections in snake-food animals since most of the identified isolates belonged to Cryptosporidium species, some of them with zoonotic potential, considered specific for rodents and resulting from ingestion of infected preys. The reptilian-specific species Cryptosporidium serpentis was identified in only one isolate. The common presence of reptile non-specific and, in some cases, zoonotic Cryptosporidium oocysts in snake faeces should to be taken into consideration in order to avoid the misidentification of the protozoan as well as the possible public health implications.

Canine leishmaniasis surveillance in a northern Italy kennel.

An epidemiological survey on canine leishmaniasis (CanL) was performed during a 3-year period (2007-2009) in a public kennel of the Bologna province. The presence of the disease was shown in the canine population for the first time in 2007 by indirect fluorescent antibody test (IFAT). The parasite circulation was confirmed also by direct diagnostic tools, as PCR, cytology and cultural method, performed on different bioptic materials. The parasite was isolated and identified as Leishmania infantum zymodeme MON 1. The serological monitoring was performed also in 2008 and 2009 on animals that previously showed negative or uncertain results. The incidence values calculated by significant seroconversions in IFAT titre ≥ 1/160, ranged between 4.9% and 6.6%, indicating a stable focus of leishmaniasis. The entomological survey, performed by sticky and CO(2)-baited traps in 2008, showed the presence of the vector Phlebotomus perfiliewi. This study allowed us to identify a stable focus of CanL in an area that was not considered eco-compatible with the presence of the vector and infection. Our results confirm the northward spread of CanL towards areas not previously affected by autochthonous foci.

Spirocerca lupi isolated from gastric lesions in foxes (Vulpes vulpes) in Sicily (Italy).

Spirocerca lupi (Rudolphi 1809) is a cosmopolitan nematode of dogs and wild carnivores. In the past it has been reported in Italy, mainly in southern regions and in Sicily, where the parasite was observed in foxes in 2005. The parasite typically produces nodular masses in the oesophagus and thoracic aorta. During the 2003-2004 hunting season, the authors investigated a total of 55 foxes (Vulpes vulpes) hunted or killed by car accidents in the provinces of Palermo and Agrigento. All the foxes were subjected to necropsy and 6 (9.16%) had S. lupi nodules located exclusively in the gastric wall. The nature of the nodules was determined by opening them and detecting the nematodes inside, which were identified as S. lupi. Some of the nodules were characterized anatomopathologically and histopathologically. The formation of the parasitic nodule in the stomach only suggests a deviation from the route commonly followed by the nematode to reach the oesophagus, the elective anatomical site for completion of its lifecycle. This survey gives a contribution to the epidemiology of this parasite which is severely outdated in Italy and highlights some distinctive features of the life cycle and parasite migration.

Spatial niche competition among helminth parasites in horse's large intestine.

The spatial distribution of large intestinal helminth parasites of 50 horses was studied. Both adult and mucosal larval stages were examined and counted within the ventral colon, dorsal colon and caecum. Thirty-three species of adult parasites were identified and their distribution in the three large intestinal regions reported, together with the localization of cyathostome encysted larvae and of empty mucosal cysts. In order to highlight interspecific interactions, both Spearman pairwise correlation (among all parasites) and multivariable negative binomial regression (among the most abundant parasites) were performed. Interactions among parasite species are very important from the ecological and parasitological point of view as they can modulate the abundance and the spatial distribution of parasites, as well as the composition of the infracommunity. The main results were: the selection of preferred sites of horse's large intestinal helminths has been confirmed; all statistically significant pairwise correlation coefficients (359 out of 666) were positive; the build up of multivariable regression models brought to light both negative and positive interactions; the most interesting competitive interactions have been observed between the most abundant and relatively large strongyle species: Strongylus edentatus and Strongylus vulgaris in the caecum and ventral colon. When S. edentatus is in the caecum, the favourite site of S. vulgaris, the latter decreases especially in the caecum. On the other hand, when S. edentatus is in the ventral colon, its favourite site, there is no negative relationship with S. vulgaris in the ventral colon and the positive correlation observed with bivariate analysis is maintained. The common belief that the horse large intestinal parasite community is isolationist, due to the absence of negative pairwise interaction, is therefore contradicted by the present study.

Influence of gastrointestinal trichostrongylidosis on ram fertility.

A study on the influence of gastrointestinal trichostrongyles on ram fertility was performed. Two groups of semen donor rams (Sarda breed) were utilized; one was experimentally infected with Haemonchus contortus, Trichostrongylus colubriformis and Teladorsagia circumcincta and the other was used as a control group. In all animals, coprological, haematological and reproductive parameters were studied. The results suggest that the parasites had a limited effect with some changes in phosphorus, cholesterol and chlorine levels. In our experience the parasitic burden produced no significant change on ram fertility.

Coccidiosis of wild and captive European mouflons (Ovis aries) living in a natural reserve of central Italy.

Between December 2005 and November 2006, a survey on coccidiosis of wild and captive mouflons living in the Orecchiella Natural Reserve (Tuscany, Italy) was performed on faecal samples collected approximately every two months. A total of 96 samples were collected, 55 of which from wild mouflons and 41 from captive mouflons. On these faecal samples qualitative and quantitative parasitological analyses were performed; moreover, faecal cultures were made in order to identify the involved Eimeria species. Significantly higher prevalence (%) and mean intensity (mean OPG +/- SD) of coccidian infection resulted for captive mouflons (73.17% and 814.6 +/- 1297.2 OPG) comparing to wild mouflons (36.73% and 112.7 +/- 268.7 OPG). Eimeria parva, E ovinoidalis, E. bakuensis, E. ahasata, E. intricata, E. crandallis, E. granulosa, E. faurei and an Eimeria species very similar to the descriptions previously reported for E. arloingi and E. yakimoffmatschoulsky, were isolated.

Cystic echinococcosis in equids in Italy.

Between March 2003 and February 2007, the livers and the lungs of 2,231 horses from various Italian regions were examined for cystic echinococcosis presence at the time of slaughter. Hydatid cysts were found in six horses, namely four from Sardinia, one from Sicily, and one from Tuscany. The location, number, morphology, and fertility of the cysts found were determined. DNA was extracted from the germinal layers and protoscoleces of the fertile cysts and polymerase chain reactions (PCR) were performed in order to strain type DNA isolates for reduced nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1), cytochrome c oxidase subunit 1 (CO1) and 12S partial genes. The PCR products were then purified and sequenced in forward and reverse. Hydatid materials obtained from positive animals were identified as Echinococcus granulosus s.s. (old G1, sheep strain) and Echinococcus equinus (old G4, horse strain) for ND1, CO1, and 12S partial genes. This allowed us to record the presence of the E. equinus in Italy for the first time with molecular tools and also to report new data on the epidemiological situation of this parasite in Italy.

Efficacy of oxfendazole and fenbendazole against tortoise (Testudo hermanni) oxyurids.

Thirty-six tortoises (Testudo hermanni) with naturally acquired oxyurids infections were used to assess the anthelmintic efficacy of oxfendazole (Dolthene; Merial) and fenbendazole (Panacur; Hoechst Roussel Vet). Animals were randomly assigned to three groups (A, B, and C) based on sex and weight. Animals in group A (seven males and six females) were orally treated with oxfendazole at dose rate of 66 mg/kg, group B animals (nine males and eight females) were orally treated with fenbendazole at dose rate of 100 mg/kg, and group C animals (three males and three females) were not treated and served as controls. All animals were individually stabled in plexiglas boxes under controlled conditions of temperature, humidity, and light beginning 7 days pretreatment and continuing for the duration of the trial. Individual tortoises feces were examined daily by the McMaster technique and drugs efficacy was assessed by the fecal eggs count reduction (FECR) test. Both drugs showed 100% of FECR. However, oxfendazole reached this level 12 days after treatment, whereas 31 days after treatment were necessary to obtain the same stable result with fenbendazole. The two drugs were well tolerated by all the animals and no adverse reactions were observed after treatment.

[An insight into the epidemiology and economic impact of gastro-intestinal nematodes in small ruminants]. / Nematodi gastrointestinali dei piccoli ruminanti: note di epidemiologia ed impatto economico.

Milk production has been a distinguishing characteristic of sheep breeding since antiquity in the Mediterranean area, including Italy, where prehistoric findings confirm this particular husbandry activity. In Homer's Odyssey, the description of Polifemo's sheep flock organization is remarkably similar to current production systems. The ancient roots of this tradition have grown into an important economic reality, with excellent levels of milk production and apporoximately 65 typical cheeses. It is interesting to note that the Italian word "pecora" (sheep) is believed to have originated from the Latin "pecunia" (money). Although heavy production losses are due to parasitic infections, only 250 papers have been published in the last 50 years in Italy on sheep parasites. Differences in climate, environmental factors and production tecniques may influence infection prevalence, load and pathogenesis. For this reason, prevention must be aimed at the particular epidemiological situation and not simply adapted from current schemes used abroad. The aim of this paper is to illustrate the epidemiology of ovine gastrointestinal nematodes in Italy and to evaluate the economic importance of their control.

Proposal of Health Information System (HIS) as tool for the epidemiological surveillance of leishmaniasis in urban areas.

In recent years leishmaniasis has become a serious Public Health problem in Italy. The coexistence of infected dogs, vectors and humans, makes urban areas suitable environment for the spread of this zoonotic disease. With the purpose to improve the current system for the control of leishmaniasis in urban areas, the authors report their experiences carrying out a Health Information System (HIS) for the surveillance of the disease in the city of Messina. The HIS project is built on a dataset containing geographical data on urban environment, census tracts at parish level and data of human and canine leishmaniasis cases reported in the last years. The HIS allows the spatial representation of the disease occurrence and can be used as a novel epidemiological tool to perform area-based surveillance.

Urban faecal pollution and parasitic risk: the Italian skill.

Owing to the recent emergency of urban faecal pollution it is important to point out new strategies in order to fight the problem starting with the knowledge of past experiences. A punctual chronography of the different approach (telluric, coprological--zoonotic and phenomenological) was attempted to furnish a scientific base for future, modern approach.

Seasonal 4-year investigation into the role of the alpine marmot (Marmota marmota) as a carrier of zoophilic dermatophytes.

Two hundred and six samples of alpine marmot (Mamota marmota) hair (148 from adults and 58 from young subjects), 102 soil samples from the entrances to the burrows of the above individuals and 20 control specimens (obtained from adjoining areas away from the burrow systems where the rodents are not usually present) were examined from May 1994 to September 1997. Seventy-five isolates belonging to six species of dermatophytes were found in 69 of the 206 hair samples examined (33.5%). Two species were zoophilic, Microsporum canis (7.8%) and Trichophyton mentagrophytes (11.2.%), and four geophilic, Microsporum cookei (2%), M. gypseum (5.8%), Trichophyton ajelloi (3.9%) and T. terrestre (5.8%). The prevalence of each species in the hair samples did not change significantly according to year, season (chi-squared test [limit significance: P <0.05] gives no significant values [P>0.05] both in year and in season comparison) or age/sex (adult versus juvenile: P=0.1; male versus female: P=0.8) of the marmot. Twenty-three of the 102 soil samples (22.5%) were positive for dermatophytes found in the hair of marmots from the same burrow systems. Five of the 20 control soil samples (25%) were positive for dermatophytes. One isolate of M. gypseum, three of T. terrestre and one of T. mentagrophytes were obtained. Compared with other free-ranging rodent hosts studied in Europe, this mycoflora is characterized by the presence and relatively high prevalence of M. canis, frequently reported in symptomatic and asymptomatic cats, dogs and fur animals. M. canis has not been isolated in other rodents in the wild. However, it has recently been reported in asymptomatic foxes (Vulpes vulpes) from northern Italy. The close link between V vulpes and M. marmota, with the former representing the most important mammal predator of the latter in the Alps (only a fraction of the predator's attacks result in the death of the rodent) may have favoured the adaptation of M. canis to this rodent host. The stable character of the M. canis/M. marmota relationship (no seasonally or annually related difference in the prevalence of this dermatophyte has been found) suggests the inclusion of the alpine marmot in the reservoir of this zoophilic pathogenic agent. In this situation, hibernation in labyrinthine burrow systems, where temperature and moisture ranges are quite uniform the whole year round, may favour the viability of M. canis arthroconidia, whose survival in mountain habitat might otherwise be compromised. This seems to be confirmed by the fact that the fungus has never been found in the control samples collected at a distance of 300 m from the outer edge of the sampled burrow systems.