Maria Teresa Manfredi

Ticks infesting humans in Italy and associated pathogens.

BackgroundTicks may transmit a large variety of pathogens, which cause illnesses in animals and humans, commonly referred to as to tick-borne diseases (TBDs). The incidence of human TBDs in Italy is underestimated because of poor surveillance and the scant amount of studies available.MethodsSamples (n = 561) were collected from humans in four main geographical areas of Italy (i.e., northwestern, northeastern, southern Italy, and Sicily), which represent a variety of environments. After being morphologically identified, ticks were molecularly tested with selected protocols for the presence of pathogens of the genera Rickettsia, Babesia, Theileria, Candidatus Neoehrlichia mikurensis, Borrelia and Anaplasma.ResultsTicks belonged to 16 species of the genera Argas, Dermacentor, Haemaphysalis, Hyalomma, Ixodes and Rhipicephalus, with Ixodes ricinus (59.5%) being the species most frequently retrieved, followed by Rhipicephalus sanguineus sensu lato (21.4%). Nymphs were the life stage most frequently retrieved (41%), followed by adult females (34.6%). The overall positivity to any pathogen detected was 18%. Detected microorganisms were Rickettsia spp. (17.0%), Anaplasma phagocytophilum (0.8%), Borrelia afzelii (0.5%), Borrelia valaisiana (0.3%), C. N. mikurensis (0.5%) and Babesia venatorum (0.6%).ConclusionsResults indicate that people living in the Italian peninsula are at risk of being bitten by different tick species, which may transmit a plethora of TBD causing pathogens and that co-infections may also occur.

Echinococcus ortleppi and E. granulosus G1, G2 and G3 genotypes in Italian bovines

To increase the knowledge on Echinococcus genotypes infesting cattle and water buffaloes (Bubalus bubalis) born and bred in Italy, the germinal layer of hydatid cysts was collected from the liver and the lungs of 80 animals slaughtered in 2007. Two mitochondrial genes (the cytochrome c oxidase subunit I and the NADH subunit I) were tested by PCR. Four genotypes were identified: G1 (sheep strain), G2 (Tasmanian sheep strain), G3 (buffalo strain), and G5 (cattle strain). Fertile cysts were detected only in the lungs of 4.5% of the total G1 lung cysts, of 9.4% of the total G3 lung cysts, and in the only G5 infected animal. This is the first report of Echinococcus ortleppi (genotype G5) in Italy.

Distribution of Anisakis simplex in fish caught in the Ligurian Sea.

Abstract A total of 822 individuals of 5 fish species were sampled in the western coastal area of the Mediterranean Sea and examined for the presence of Anisakis simplex larvae in the body cavity, muscle, and viscera. A. simplex mean intensity (mI) and prevalence (Pr) values were very high in the common scad (Trachurus trachurus; Pr 80–100%, mI 19.3–36.8) and the blue scad (T. picturatus; Pr 80–100%, mI 18.2–70.7), moderate in the blue whiting (Micromesistius poutassou), and low in the red mullet (Mullus barbatus) and the striped mullet (M. surmuletus). The larval burden was strongly related to host length in the common and blue scad.

Host specificity of abomasal nematodes in free ranging alpine ruminants.

Abomasums from 641 alpine wild ruminants representing five different species (Cervus elaphus, Capreolus capreolus, Rupicapra rupicapra, Capra ibex, Ovis musimon) and from 19 domestic sheep (Ovis aries) from alpine areas were examined in order to investigate the host-specificity of abomasal helminths. Nine out of 20 helminth species were found in at least five different host species. A discriminant analysis was able to significantly discriminate the hosts on the basis of their helminth community composition with the exception of O. musimon and O. aries. Based on the correlation between each variable represented by helminth species with the most explanatory discriminant axis, it was possible to classify helminths into specialists and generalists. Specialists are represented by the dominant species in a community of an host species or family while generalists appear in the communities of many different hosts as intermediate species. Due to the pathogenic potential of some of these generalist species (i.e. Haemonchus contortus, Trichostrongylus axei) and their ability to adapt easily to the conditions found in several different hosts, they appear to be the most important from a sanitary point of view.

Intestinal Parasites of Owned Dogs and Cats from Metropolitan and Micropolitan Areas: Prevalence, Zoonotic Risks, and Pet Owner Awareness in Northern Italy

Intestinal parasites of dogs and cats are cosmopolitan pathogens with zoonotic potential for humans. Our investigation considered their diffusion in dogs and cats from northern Italy areas, specifically the metropolitan area of Milan and two micropolitan areas of neighboring provinces. It included the study of the level of awareness in pet owners of the zoonotic potential from these parasites. A total of 409 fresh fecal samples were collected from household dogs and cats for copromicroscopic analysis and detection of Giardia duodenalis coproantigens. The assemblages of Giardia were also identified. A questionnaire about intestinal parasites biology and zoonotic potential was submitted to 185 pet owners. The overall prevalence of intestinal parasites resulted higher in cats (47.37%−60.42%) and dogs (57.41%−43.02%) from micropolitan areas than that from the metropolis of Milan (dogs: P = 28.16%; cats: P = 32.58 %). The zoonotic parasites infecting pets under investigation were T. canis and T. cati, T. vulpis, Ancylostomatidae, and G. duodenalis assemblage A. Only 49.19% of pet owners showed to be aware of the risks for human health from canine and feline intestinal parasites. Parasitological results in pets and awareness determination in their owners clearly highlight how the role of veterinarians is important in indicating correct and widespread behaviors to reduce risks of infection for pets and humans in urban areas.

Gastrointestinal nematodes of dairy goats, anthelmintic resistance and practices of parasite control in Northern Italy

BackgroundGastrointestinal nematodes (GINs) are one of the main constraints to ruminant production worldwide. Anthelmintic resistance (AR) has been reported in goats throughout Europe, yet little is known about the AR status in Italy. The aims of the study were: i) determine the frequency of AR in GINs in goat flocks in Northern Italy, Italy, ii) survey goat farmers on the current practices of parasite control, iii) update the species composition of the gastrointestinal helminthofauna. Thirty three flocks were enrolled and 1288 individual fecal samples were collected. Based on the egg per gram (EPG), 15 flocks were selected to evaluate the presence of AR in GINs with the Fecal Egg Count Reduction Test (FECRT). A questionnaire surveyed 110 dairy goat farmers to acquire information about farm management and drenching practices against GINs. Further, the gastrointestinal tracts of 42 goats were analyzed.ResultsThe FECRs indicated that five of the 15 flocks had problems of AR, which was identified in all two of the anthelmintic classes tested. Resistance and suspected resistance was found in 40% of the flocks selected for AR testing that were treated with benzimidazoles while 20% of the flocks treated with eprinomectin had resistant GINs. Teladorsagia/Trichostrongylus L3 were isolated from the post-treatment coprocultures of all flocks with resistance but not from the flock with suspected oxfendazole resistance. Treatments against helminths were performed once annually in 73.63% of the flocks, but 20.00% of farmers declared not regularly treating their goats every year. Annual treatments usually occurred in autumn or winter at dose rate for sheep. Te. circumcincta, H. contortus, Tr. colubriformis, Skrjabinema caprae and Oesophagostomum venulosum were the most abundant and prevalent species of the gastrointestinal tract.ConclusionsStrategies to prevent the development of AR should be widely adopted in Northern Italy. Further, farmers and practitioners should be educated about the importance of using the correct dose rates in goats. In addition, the presence of highly pathogenic GINs coupled with high worm burden in all sector of gastrointestinal tract and the prevalence values further suggest that improved diagnosis and active surveillance of GINs infection is needed.

High-resolution phylogeography of zoonotic tapeworm Echinococcus granulosus sensu stricto genotype G1 with an emphasis on its distribution in Turkey, Italy and Spain.

Echinococcus granulosus is the causative agent of cystic echinococcosis. The disease is a significant global public health concern and human infections are most commonly associated with E. granulosus sensu stricto (s. s.) genotype G1. The objectives of this study were to: (i) analyse the genetic variation and phylogeography of E. granulosus s. s. G1 in part of its main distribution range in Europe using 8274 bp of mtDNA; (ii) compare the results with those derived from previously used shorter mtDNA sequences and highlight the major differences. We sequenced a total of 91 E. granulosus s. s. G1 isolates from six different intermediate host species, including humans. The isolates originated from seven countries representing primarily Turkey, Italy and Spain. Few samples were also from Albania, Greece, Romania and from a patient originating from Algeria, but diagnosed in Finland. The analysed 91 sequences were divided into 83 haplotypes, revealing complex phylogeography and high genetic variation of E. granulosus s. s. G1 in Europe, particularly in the high-diversity domestication centre of western Asia. Comparisons with shorter mtDNA datasets revealed that 8274 bp sequences provided significantly higher phylogenetic resolution and thus more power to reveal the genetic relations between different haplotypes.

Seroprevalence of Toxoplasma gondii in wild boars (Sus scrofa) from Central Italy

Wild and farmed game meat consumption has been highlighted as an emerging risk factor for Toxoplasma gondii infection in humans. In Central Italy wild boar is widely distributed and is also one of the most popular game species. The main goal of the present study was to estimate the seroprevalence of T. gondii antibodies through a serological survey conducted on 400 hunted wild boars (250 males and 150 females) during three subsequent hunting seasons (2009–2011), using an Immunofluorescence Antibody Assay. The animals were sorted by age, determined on the evaluation of the dental table; 101 were <1 year old, 175 from 1 to 3 years, and 124 > 3 years. Antibodies against T. gondii were detected in 56 (14%) serum samples with titers ranging from 40 to ≥160; a significant association (p < 0.05) was found between seropositivity and age, but not gender, hunting districts, or year of sampling.

Intestinal helminth communities of the red fox (Vulpes vulpes L.) in the Italian Alps

The study analyses the intestinal helminth communities found in 645 red foxes of alpine populations from five areas of Northern Italy. In particular, the aim was to evaluate the contribution of both environmental variability and the structure of fox population (extrinsic and intrinsic factors, respectively) in shaping the structure and composition of the intestinal helminth communities. To identify the influence of the characteristics of the host (i.e. age and sex) together with extrinsic factors (year, site, season, and altitude of collection) on number of species, total load per fox, prevalence and abundance of each parasite species, general linear models were performed. Fifteen helminth species have been detected in the intestine of 545 infected animals (84.5%) with a total of 17,144 parasites collected. The analysis of factors influencing both prevalence and abundance of infection of the parasite species revealed the preponderance of extrinsic factors on intrinsic ones. In particular, geographical areas influenced prevalence and abundance of every parasite species, showing high spatial variability. The lower influence of host factors may suggest that, in this case, host dynamics play a trivial role with respect to spatial variability in determining parasite abundance. These results pointed out high prevalence of intestinal helminth infections in the Italian Alpine fox, confirmed the typical composition of parasite fauna within fox populations, underlined local differences in the structure and composition of the helminth communities. Moreover, this study highlights the major role of extrinsic factors vs intrinsic ones.

Allergenic activity of Molicola horridus (Cestoda, Trypanorhyncha), a cosmopolitan fish parasite, in a mouse model

The cestode Molicola horridus is a muscle parasite of teleost fish. The ability of molecules present in this parasite to induce allergic response is not known yet. Since fish-borne parasitic allergens can induce allergic manifestations even when the parasitized fish is well cooked, the knowledge of potential allergens present in food is important in order to provide a save products for consumers. The aim of the study was to determine the allergenic potential of the components present in the crude larval extract (CLE) of M. horridus. Two mouse models were exposed to the CLE: adult BALB/c mice that were intraperitoneally (i.p.) immunized and newborn BALB/c mice that were orally exposed. Specific antibody levels in serum and faeces were measured by ELISA. The cellular immune response was determined by proliferation assay of splenocytes from sensitized mice. The protein profile of CLE was analysed by SDS-PAGE and western blot. In adult mice, specific IgG and IgA were detected in sera and faeces, whereas specific IgE were detected in sera only. In newborn mice, specific IgG were detected in sera and a low level of IgA was detected in faeces. SDS-PAGE revealed the CLE protein profile, with most of the proteins running from 15 to 50kDa. Specific IgG recognized mainly the 26 and 75kDa proteins and a molecular complex below 100kDa by immunoblot. Specific IgE recognized the same 26kDa protein as IgG did, and, with less intensity, another protein at 30kDa. Splenocytes from CLE-immunized mice proliferated when stimulated with CLE in a dose-dependent manner. The crude larval extract from M. horridus has potential allergenic molecules which can represent a risk for fish consumers.