Mario Pietrobelli

The northward spread of leishmaniasis in Italy: evidence from retrospective and ongoing studies on the canine reservoir and phlebotomine vectors

Visceral leishmaniasis (VL) incidence has been increased in Italy in humans and dogs since the 1990s, with new foci being detected within traditional boundaries of endemic transmission but also in northern regions previously regarded as non‐endemic. To monitor the putative VL spreading, surveillance was implemented in northern continental Italy comprising: analysis of human cases recorded from 1990 through 2005; retrospective literature analysis of canine leishmaniasis (CanL) and phlebotomine sandfly records through 2002; prospective investigations in dogs from 2003 through 2005 and surveys on sandflies in 2003 and 2004. Two‐hundred‐thirty human cases (11% of Italian cases) were recorded. Their stratification by age and HIV status disclosed a sharp decrease of HIV/VL co‐infections paralleled by concomitant increase of paediatric and HIV‐negative adult patients during the study period. Four patients had no travel history. Seven leishmaniasis foci were retrospectively identified since 1990, whereas prospective investigations in dogs disclosed 47 autochthonous clinical cases and 106 autochthonous seropositives among 5442 dogs (2.1%) from 16 foci of six regions. Parasites were typed as Leishmania infantum MON‐1. Four vector species were identified among 1696 Phlebotomus (Larroussius) collected specimens. Comparisons with historical data showed that P. perniciosus and P. neglectus have increased in density and expanded their geographic range in the study area. Northern continental Italy is now focally endemic for VL and a moderate risk for human disease does exist, although the intensity of transmission seems to be lower than in traditional settings of Mediterranean VL.

Prevalence and diagnosis of Babesia and Theileria infections in horses in Italy: a preliminary study.

Babesia caballi and Theileria equi are the causative agents of equine piroplasmosis. In this preliminary epidemiological study, 412 horses reared in central and northern Italy were sampled and three diagnostic methods compared, namely, the microscopy, the indirect fluorescent antibody test (IFAT) and a PCR. Possible risk factors (such as area, season, breed, activity, sex, age, and grazing) associated with serological positivity were evaluated. A seroprevalence of 68.4% was found: 12.4% of the animals had anti-T. equi antibodies, 17.9% anti-B. caballi antibodies and 38.1% had antibodies against both species. Of the seropositive samples, 3.1% and 9.4% were positive to microscopy and PCR, respectively; 31.5% of the horses were IFAT-negative but 1.4% and 2.4% of the corresponding blood samples were positive to microscopy and PCR, respectively. Molecular techniques revealed that the species present were closely related to T. equi, Theileria sergenti, Theileria buffeli and the Babesia microti-like piroplasm provisionally named Theileria annae. Grazing was found to be a pronounced risk factor for equine piroplasmosis.

Tick reservoirs for piroplasms in central and northern Italy.

Ticks, collected in central and northern Italy from pets, livestock, wild animals and the environment (n=2107), were identified by microscopy and processed by molecular diagnostics to determine the species that act as a reservoir for piroplasms. A total of 11 ixodid tick species were identified, with five of them proving to be piroplasm positive. Molecular diagnostics identified Theileria equi and eight Babesia species in 52 adult specimens, mostly (n=50) removed from piroplasm-free vertebrate hosts. Ixodes ricinus hosted the highest number of species, although the highest infection rate was recorded in Hyalomma marginatum (9.1%), followed by I. ricinus (5.1%), Dermacentor marginatus (5%), Rhipicephalus turanicus (3.1%) and R. sanguineus (1.2%). Novel tick/pathogen associations were detected, suggesting that certain tick species (such as Hy. marginatum, R. sanguineus and I. ricinus) are vector of more piroplasm species than previously thought. Trans-stadial maintenance of the piroplasms was observed in each positive tick species; vertical transmission of B. canis canis was demonstrated in R. sanguineus. Finally, the detection of Babesia sp., B. microti-like species and B. rodhaini, phylogenetically related to zoonotic species, suggests that the human population could be at risk of infection in the studied area.

Environmental contamination by canine geohelminths

Intestinal nematodes affecting dogs, i.e. roundworms, hookworms and whipworms, have a relevant health-risk impact for animals and, for most of them, for human beings. Both dogs and humans are typically infected by ingesting infective stages, (i.e. larvated eggs or larvae) present in the environment. The existence of a high rate of soil and grass contamination with infective parasitic elements has been demonstrated worldwide in leisure, recreational, public and urban areas, i.e. parks, green areas, bicycle paths, city squares, playgrounds, sandpits, beaches. This review discusses the epidemiological and sanitary importance of faecal pollution with canine intestinal parasites in urban environments and the integrated approaches useful to minimize the risk of infection in different settings.

Wild Ungulates as Babesia Hosts in Northern and Central Italy

Babesia and Theileria species were investigated in wild ungulates of Northern and Central Italy. Of 355 blood samples examined, 108 (30.4%) were positive to molecular diagnostics (polymerase chain reaction [PCR] with specific primers and sequencing). The sequence analysis showed that the roe deer is a susceptible host for several piroplasms belonging both to Babesia (31%) and Theileria (14.2%) species, whereas fallow deer and wild boar harbor only Theileria species (49% and 2.6%, respectively). Strains related to B. divergens are highly present (28.3%) in the roe deer, which, however, also harbors Babesia MO1 type and Babesia microti-like organisms. Babesia EU1 type is described for the first time in a roe deer in Italy. The finding in roe deer of Babesia species involved in human babesiosis is of concern for public health, mainly because ecological changes in progress cause the increase of both the deer species and the vector tick populations.

Prevalence of Giardia spp. in dogs and humans in northern and central Italy.

Despite the public health importance and the possible zoonotic role of Giardia, epidemiological survey on the prevalence of giardiosis in Italy are lacking and fragmentary, compared to other countries. A cross-sectional coprological study on giardiosis was carried out in central and northern Italy in dogs and humans with the aim to providing data about prevalence and potential risk factors. A total of 616 dogs (436 from central and 180 from northern Italy) and 300 humans (central Italy) were examined. The overall prevalence was 21.3% in dogs and 2% in humans. No differences were noted between dog’s prevalence in central and northern Italy (22.2% and 19.4% respectively). The major risk factors for giardiosis in dogs were represented by density of dogs and age, i.e. dogs kept in kennel (PR=2.13, APe=53%) and younger than 1 year (PR=1.99, APe=50%) showed higher prevalence than others. Higher prevalence was noted also in dogs with gastrointestinal symptoms (PR=1.86%, APe=46%). Looking forward to obtain information on these Giardia genotypes, all the measures for the prevention of giardiosis, screening of dogs for giardiosis, particularly puppies, even those with no symptoms, and their treatment to prevent eventual human infection is highly recommended.

Giardia species and other intestinal parasites in dogs in north-east and central Italy

Faecal samples were collected from dogs during the period from 2001 to 2003, in seven provinces of two Italian regions: Veneto, in the north-east, and Abruzzi, in central Italy. The minimum sample size required (246 dogs) was calculated according to Thrusfield (1995) considering the population size (infinite), expected prevalence (20 per cent), confidence interval (

Assessment of cattle grub (Hypoderma spp.) prevalence in northeastern Italy: an immunoepidemiological survey on bulk milk samples using ELISA.

Hypodermosis is a myiasis of cattle mainly found in the Northern Hemisphere. In Italy it has been reported primarily in southern regions with a farm prevalence reaching a peak of 85%, while few data are available in the north of the country. This paper reports the results of a serological survey carried out on hypodermosis in Vicenza province (northeastern Italy). During January and February 1999, bulk milk samples were collected from 224 dairy farms and tested, using an hypodermosis ELISA kit. Epidemiological data, such as farm location, herd size, number of cows, grazing management, treatments against Hypoderma larvae, presence of imported animals, were gathered and statistically analysed in relation to the serological results. In 15 farms the presence of warbles was monitored by manual palpation and larvae collected for identification.Ninety-seven (43.3%) samples were detected as positive, whereas 12 (5.4%) and 115 (51.3%) were inconclusive and negative, respectively. Thirteen farms were positive upon parasitological examination and the larvae collected were identified as Hypoderma bovis.Seroprevalence was highly correlated with the practice of free grazing and slightly, but significantly, correlated with the number of cows on the farm. The mean antibody level of positive herds was significantly higher on farms without imported animals. On farms with warbled animals there was a high inverse correlation between antibody level and quantity of milk in the tanks and the number of lactating cows. The ELISA test on bulk milk samples has been shown, under field conditions, to be a useful, cost-effective and easy test for the diagnosis of bovine hypodermosis.

Investigating the Determinants of Toxoplasma gondii Prevalence in Meat: A Systematic Review and Meta-Regression

Background Toxoplasma gondii is one of the most widespread parasites in humans and can cause severe illness in immunocompromised individuals. However, its role in healthy people is probably under-appreciated. The complex epidemiology of this protozoan recognizes several infection routes but consumption of contaminated food is likely to be the predominant one. Among food, consumption of raw and undercooked meat is a relevant route of transmission, but the role of different meat producing animal species and meats thereof is controversial. Objectives The aim of the present work is to summarize and analyse literature data reporting prevalence estimates of T. gondii in meat animals/meats. Data Sources We searched Medline, Web of Science, Science Direct (last update 31/03/2015). Eligibility Criteria Relevant papers should report data from primary studies dealing with the prevalence of T. gondii in meat from livestock species as obtained through direct detection methods. Meta-analysis and meta-regression were performed. Results Of 1915 papers screened, 69 papers were included, dealing mainly with cattle, pigs and sheep. Pooled prevalences, based on random-effect models, were 2.6% (CI95 [0.5–5.8]) for cattle, 12.3% (CI95 [7.6–17.8]) for pigs and 14.7% (CI95 [8.9–21.5]) for sheep. Due to the high heterogeneity observed, univariable and multivariable meta-regression models were fitted showing that the geographic area for cattle (p = 0.032), the farming type for pigs (p = 0.0004) and the sample composition for sheep (p = 0.03) had significant effects on the prevalences of Toxoplasma detected/estimated. Moreover, the role of different animal species was dependent on the geographic location of animals’ origin. Limitations Limitations were due mainly to a possible publication bias. Conclusions and Implications The present work confirms the role of meat, including beef, as T. gondii sources, and highlights the need for a control system for this parasite to be implemented along the meat production chain. Moreover, consumer knowledge should be strengthened in order to reduce the impact of disease.

Serological assays on eight cases of human dirofilariasis identified by morphology and DNA diagnostics

Specimens of Dirofilaria sp. removed from eight Italian patients were identified as D. repens by morphology and confirmed as such by a PCR-based method of DNA analysis. Blood samples were also drawn from the patients so that two serological tests (ELISA and western blot), one based on the recognition of molecular markers recently identified in the somatic antigenic complex of D. repens, could be evaluated. The antigenic complex used in the ELISA only gave a weak sensitivity. However, the western-blot assays, based on the polypeptide molecular markers, were found to have greater sensitivity and should be useful in detecting human cases of dirofilariasis.