Sergio Aurelio Zanzani

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.

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.

Canine Fecal Contamination in a Metropolitan Area (Milan, North-Western Italy): Prevalence of Intestinal Parasites and Evaluation of Health Risks

Intestinal parasites of dogs represent a serious threat to human health due to their zoonotic potential. Thus, metropolitan areas presenting high concentrations of pets and urban fecal contamination on public areas are at sanitary risk. Major aim of this survey was to determine prevalence of zoonotic parasites in dog fecal samples collected from public soil of Milan (north-western Italy). Differences in parasites prevalence distribution were explored by a geographical information system- (GIS-) based approach, and risk factors (human density, sizes of green parks, and dog areas) were considered. The metropolitan area was divided into 157 rectangular subareas and sampling was performed following a 1-kilometer straight transect. A total of 463 fecal samples were analyzed using centrifugation-flotation technique and ELISA to detect Giardia and Cryptosporidium coproantigens. A widespread fecal contamination of soil was highlighted, being fecal samples found in 86.8% of the subareas considered. The overall prevalence of intestinal parasites was 16.63%. Zoonotic parasites were found, such as Trichuris vulpis (3.67%), Toxocara canis (1.72%), Strongyloides stercoralis (0.86%), Ancylostomatidae (0.43%), and Dipylidium caninum (0.43%). Giardia duodenalis was the most prevalent zoonotic protozoa (11.06%), followed by Cryptosporidium (1.10%). Faeces from subareas characterized by broad green areas showed to be particularly prone to infection.

Besnoitia besnoiti among cattle in insular and northwestern Italy: endemic infection or isolated outbreaks?

BackgroundBovine besnoitiosis, caused by the apicomplexan Besnoitia besnoiti, is a chronic and debilitating disease considered as emerging in Europe. In Spain, Portugal and France it is endemic and foci of infection were recorded in Germany, Switzerland, Hungary, Greece and Italy. In Italy, cases of bovine besnoitiosis were registered both in imported and autochthonous cattle, and mostly in central regions; high seroprevalence was also revealed by an epidemiological survey performed in the southern part of the country. Aiming to update information on the disease in northwestern and insular areas of Italy, where data on bovine besnoitiosis were missing, a serosurvey was designed for the present study.MethodsThree thousand one hundred and forty bovine blood samples from both dairy and beef farms (n = 126) were collected in northwestern regions (Lombardy, Piedmont and Liguria) and in the island of Sardinia. Samples were analyzed by a standardized in-house ELISA and those resulted positive were re-tested by Western Blot (WB) for confirmation. On results obtained by both ELISA and WB, apparent (AP) and true prevalence (TP) were calculated at individual and herd levels. Further, a panel of sera resulted positive to ELISA was analyzed by IFAT.ResultsA total of 712 animals (AP = 22.7%; TP = 18.8%) and 109 farms (AP = 86.5%; TP = 88.2%) showed a positive reaction in ELISA. Only ten (AP = 0.3%; TP = 0%) specimens proceeding from five farms (AP = 3.9%; TP = 1.7%) from Lombardy were confirmed positive to the WB, corresponding to two Holstein Friesian cows and eight beef cattle. IFAT showed a low sensitivity (44.4%) scoring positive in only four samples out of 9 positive to WB.ConclusionsThe survey demonstrated that bovine besnoitiosis cannot still be considered endemic in whole Italy. In fact, independent foci of infection were registered only in Lombardy region. Therefore, a sanitary strategy aimed to increase control measures and to organize monitoring plans, by adequate diagnostic tools is necessary to avoid overestimation of B. besnoiti in Italy.

Toxoplasma gondii in small ruminants in Northern Italy – prevalence and risk factors

OBJECTIVE The aim of the survey was to evaluate Toxoplasma gondii seroprevalence in small ruminants and possible risk factors associated with the infection. MATERIALS AND METHODS Sera from 474 goats and 502 sheep reared on 42 farms in northern Italy were collected and tested for IgG antibodies to T. gondii by IFAT (indirect immunofluorescence antibody test). To identify risk factors, a binary logistic regression analysis of the variables was performed. An audit form about farm management was used. RESULTS Antibodies to T. gondii were found in 96.6% of goat farms and in 87.5% of sheep farms; 41.7% goats and 59.3% sheep resulted positive. Seroprevalence was significantly higher in sheep than in goats. Seroprevalence values were similar in goats from eastern and western areas, whereas goats from the southern area were at lower risk of infection. Saanen goats presented the lowest seroprevalence (30.7 %), whereas cross-breed exhibited the highest rate (48.7%). Goats from farms housing both sheep and goats had an infection risk 1.39 times higher than goats from farms that did not house sheep. Animals bred on intensive farms showed lower prevalence (22.1%) in comparison with those from extensive (45.6%) or semi-intensive farms (60%). Sampling area was one of the strongest predictors of T. gondii infection in sheep flocks. Transhumant flocks showed a higher risk of infection by T. gondii compared with semi-intensive farms (66.8% vs. 38.4%). CONCLUSIONS The highest T. gondii seroprevalence values were registered in transhumant flocks of sheep and in family businesses rearing goats. As these traditional activities represent an important resource for the conservation of the territory and its economy, management practices for a better control of the disease should be improved.

Prevalence of echinococcosis in humans, livestock and dogs in northern Italy

SummaryThe presence of Echinococcus sp. cysts was investigated in 822 sheep, 123 goats and 112,521 cattle from Lombardy region, North Italy. Faecal samples from 40 sheepdogs were also analyzed, with 9 samples containing taeniid eggs (22.5 %), 8 samples being coproantigen-positive (20 %), and one dog from a northern province (Lecco) positively confirmed by PCR. Cystic Echinococcosis (CE) was detected in 0.36 % of sheep and in 0.29 % of cattle in 2004. No goat resulted to be infected. Data from CE patients treated in Lombardy were collected by inspecting hospital discharge records. In 2004, 156 CE-related admissions (62 % male and 38 % female) were reported in Lombardy. Total hospital stay was 1,372 days (1,286 for inpatients, 86 for outpatients). Most patients (72.4 %) were residents in Lombardy and 1.9 % were from Piedmont; the remaining patients were from central and southern Italy. According to acquired data CE resulted hypoendemic in animals in Lombardy. Prevalence rates in humans were higher than expected in this region, usually considered as non-endemic. Assessment of the prevalence of CE in humans remains a difficult, costly, time-consuming and labourintensive task. The present study suggests establishing a National Registry of Cystic Echinococcosis with the aim to highlight regional risk factors and to benefit from its matching both clinical and epidemiological data.

Study of the gastrointestinal parasitic fauna of captive non-human primates (Macaca fascicularis)

The aim of this study was to examine helminths and protozoans in cynomolgus macaques (Macaca fascicularis) imported from registered breeding facilities in China and their relation to health risks for non-human primate handlers in biomedical research centers and in breeding facilities. Fresh fecal samples were collected from a total of 443 M. fascicularis and analyzed by copromicroscopical analysis, immunoenzymatic, or molecular assays. As to helminths, whose eggs were shed in 2.03% of the samples, Trichuris and Oesophagostomum were the only two taxa found, with low prevalence and low eggs per gram (EPG) values. Protozoans were more frequently detected (87.40%), with Entamoeba coli (85.19%) and Endolimax nana (79.26%) as the most prevalent species shed. Other parasites found by fecal smear examination were uninucleated-cyst-producing Entamoebas (78.52%), Iodamoeba bütschlii (42.96%), and Chilomastix mesnili (24.44%), while cysts of Balantidium coli (22.2%) were only observed by sedimentation. No coproantigens of Giardia duodenalis, Cryptosporidium spp., and Entamoeba histolytica complex were detected. Blastocystis sp. infection was noticed in 87.63% of macaques by PCR. These cynomolgus monkeys were infected with many subtypes (ST1, ST2, ST3, ST5, and ST7), where the predominant Blastocystis sp. subtypes were ST2 (77.5%), followed by ST1 (63.5%). Data collected confirmed the presence of potentially zoonotic parasites and a high parasite diversity, suggesting the need for appropriate and sensitive techniques to adequately control them and related health risks for handlers of non-human primates in biomedical research centers and in breeding facilities.

Cystic echinococcosis in Lombardy: epidemiological aspects and spatial analysis

SummaryA descriptive spatial analysis on the spreading of cattle cystic echinococcosis in non-endemic areas of Italy (Lombardy) was performed and the presence of clusters of hydatid disease in the studied area were evaluated. Positive cattle were from 285 out of 5708 farms (4.99 %) with significant differences among the provinces considered. Infection with Echinococcus granulosus was detected in 321 out of 112,521 dairy cattle (P = 0.29 %). CE-positive farms were distributed among 195 of the 812 municipalities included in the study (24.01 %) located in the central and south-eastern part of the region. By SaTScan, four significant clusters of infection were identified; they included 4–241 cattle farms and 4–24 positive animals. The most likely spatial cluster of 17 observed cases in 241 farms was located in the north-eastern part of Lombardy. SaTScan analysis suggest that a geographical surveillance in the area of the positive clusters occur is for and that’s could be very useful to public health service to monitoring this zoonotic disease.

Detection of Cyclospora in captive chimpanzees and macaques by a quantitative PCR-based mutation scanning approach.

BackgroundCyclospora is a protistan parasite that causes enteritis in several species of animals including humans. The aim of this study was to investigate the presence of Cyclospora in captive non-human primates.MethodsA total of 119 faecal samples from Pan troglodytes, Macaca sylvanus, Cercopithecus cephus, Erythrocebus patas, Chlorocebus aethiops and Macaca fascicularis from a wildlife animal rescue center as well as from Macaca fascicularis from an experimental primate research center were tested for the presence of Cyclospora by quantitative real-time PCR (qPCR) and single-strand conformation polymorphism (SSCP) analysis.ResultsCyclospora was detected in three Pan troglodytes (13.6%) and nine (9.3%) Macaca fascicularis.ConclusionsThe present study represents the first record of Cyclospora in captive primates in Europe, suggesting the presence of Cyclospora cayetanensis, which is transmissible to humans.