Molecular detection of Rickettsia, Borrelia, and Babesia species in Ixodes ricinus sampled in northeastern, central, and insular areas of Italy.

The aim of the present study was to provide insight into the diversity of tick-borne pathogens circulating in Italy, carried/transmitted by Ixodes ricinus, one of the most abundant tick species in the country. A total of 447 specimens sampled in five areas of northeastern, central and insular Italy were analysed by polymerase chain reaction and sequencing for the presence of rickettsiae, borreliae and babesiae. Several rickettsial species of the spotted fever group of zoonotic concern and other zoonotic pathogens were found, such as Borrelia burgdorferi s.s., Borrelia afzelii, Borrelia garinii, and Babesia venatorum. These findings confirm a wide distribution of tick-borne bacterial and protozoan species in Italy, and highlight the sanitary importance of I. ricinus, often recorded as feeding on humans

Is the Goat a New Host for the G3 Indian Buffalo Strain of Echinococcus granulosus?

Four goats bred in Central Italy (province of Rieti) revealed, in the liver, metacestodes of Echinococcus granulosus. The cysts, unilocular and fertile, were examined by microscopy and molecular diagnostics. Morphological data on the rostellar hooks are in agreement with the original description of the strain found in buffaloes and are largely compatible with those reported in Europe for cattle and humans. Specific PCR followed by DNA sequencing of the mitochondrial cox1 gene revealed for all the isolates 99.5% identity to the reference strain G3 genotype and 99.3% and 99.1% to G2 and G1, respectively. Further genetic markers (nad1 and 12S rRNA) confirmed the identity of the goat isolates to the G3 strain. This genotype, here reported for the first time in goats, proved to have a wider than previously supposed host range, therefore its relevance in human hydatidosis is expected to be more often evidenced

Malaria in children of Tshimbulu (Western Kasai, Democratic Republic of the Congo): epidemiological data and accuracy of diagnostic assays applied in a limited resource setting

BACKGROUND: The literature data on malaria in Western Kasai, DRC, are limited and inadequate. A recent molecular survey there has detected Plasmodium ovale and Plasmodium malariae as mixed infections with Plasmodium falciparum. In Tshimbulu, Western Kasai, during a humanitarian initiative designed to provide children with free preventive screening and to reduce the local high malaria death rate, accurate species identification was performed, in order to collect unambiguous epidemiological data and to evaluate the reliability of locally applied diagnostics. METHODS: Finger pricks provided fresh blood for microscopic analysis (MA), for rapid diagnostic test (RDT) and for molecular diagnostics (MD). MA and RDT were first performed by the local team and then a re-interpretation of the results (on the same slides and on RDT's taken pictures) was conducted in Italy, where MD were performed. RESULTS: The analysis was conducted on 306 children; RDT found 80.9 % as P. falciparum-positive (37.4 % as two-band positive, P. falciparum single infection). MA identified a further four children as positive to P. falciparum and six co-infections with P. ovale. The second RDT evaluation confirmed a similar infection rate (78.2 %) but interpreted as two-band positive a significantly higher share of tests (56.8 %). MA confirmed 80.0 % of the children as malaria positive and, in addition to P. falciparum, identified P. malariae (13.8 %), P. vivax (3.4 %) and P. ovale (2.4 %), and detected Babesia microti in 19 smears. MD confirmed all of the species found (Babesia microti included), classified as mono-infection with P. falciparum a rate of spots comparable to MA revision, and identified all P. ovale as Plasmodium ovale wallikeri. The RDT used locally proved 93.1 % sensitive and 92.1 % specific for P. falciparum. CONCLUSIONS: The malaria prevalence among the children and the presence of four Plasmodium species, highlighted in this study, identified a sanitary issue which proved to be more alarming than expected, as it was worsened by the unpredictable presence of P. vivax and Babesia microti (never before reported in DRC). Each diagnostic tool showed its point of weakness. Therefore, the most correct approach is by the combined use of different, locally available, diagnostic tools

Canine Babesioses in noninvestigated areas of Serbia

During the years 2012-2014, a total of 158 outdoor dogs from Pančevo and Đurđevo (northern Serbia) and Niš and Prokuplje (southern Serbia) were submitted to molecular analyses (PCR and sequencing) for canine babesioses. An overall prevalence of 21.5% was found, due to the species Babesia sp. 'spanish dog' (10.1%), B. gibsoni (5.7%), B. canis vogeli (1.9%), B. caballi (1.9%), and B. microti (1.9%). In addition, sequence analysis showed the presence of Hepatozoon canis in a dog from Niš. No significant difference between infected and noninfected dogs was found by age, sex, and place of residence, whereas there was difference regarding the presence of ticks (p<0.005) and application of preventive measures such as applying of antitick drugs/devices. Moreover, a significant difference was established by area: Dogs from Prokuplje showed infection rates (59.1%) higher than dogs from Pančevo (11.9%), Niš (4.5), and Đurđevo (where infected dogs were not found), and a different geographical distribution of the species was found. The presence of so many Babesia species and the first identification of H. canis will allow investigations on the pathogenic role played by each one and suggests entomological studies on the tick species that are more suitable vectors for each of them. Finally, the presence of so many infected dogs offers the opportunity of evaluating the hypothesis of a possible zoonotic role of babesial species affecting dogs

Canine Trypanosoma cruzi infection in the Bolivian Chaco

A cross-sectional study on Trypanosoma cruzi was carried out in 2013 to evaluate the role of dogs as possible source of infection for humans in two rural communities of the highly endemic Bolivian Chaco (Bartolo, Chuquisaca Department, n = 57 dogs; and Ivamirapinta, Santa Cruz Department, n = 48 dogs). Giemsa-stained thick and thin smears, rapid immunochromatographic test (ICT) (Chagas Quick test, Cypress Diagnostic, Belgium) and polymerase chain reaction for T. cruzi on dried blood spots were performed. All smears proved negative by microscopic examination, whereas 23/103 (22%) were positive by ICT and 5/105 (5%) blood samples contained T. cruzi DNA, evidencing the potential role of dogs in the domestic transmission of the parasite

Human oestriasis acquired in Florence and review on human myiasis in Italy

Myiases-causing flies are widely observed in tropical countries, whereas in Italy, a temperate country, their epidemiology and clinical presentation are poorly known. We report three cases of human conjunctival myiasis recently observed at our hospital, and the results of a review of the literature on human myiasis in Italy. In August 2012, a case of Oestrus ovis conjunctival myiasis acquired in the city centre of Florence, Italy was diagnosed at our hospital. In the early fall of 2013, two additional cases, acquired in the neighbouring areas, occurred. The review of literature showed that, up to the middle of 1990s, myiasis in Italy was mainly an occupational disease of shepherds, caused by O. ovis. Recently, cases of travel acquired furuncular myiasis emerged, together with “opportunistic” autochthonous cases of wound myiasis in patients with underlying health conditions. Considering the causative agents of human of myiasis in Italy, among the 703 autochthonous cases reported, 98.1 % were caused by O. ovis, while among the 42 imported cases described, 59.5 % were due to Cordylobia spp. and 40.5 % to Dermatobia hominis. Our findings suggest that O. ovis conjunctival myiasis may still be observed in urban setting in Italy. Health care providers should know and implement the basic rules of entomoprophylaxis for myiasis in the facilities where they are working and use these indications to educate patients and care givers in both pretravel care and geriatric outpatient settings. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00436-014-3906-9) contains supplementary material, which is available to authorized users

Investigation on the occurrence of Echinococcus multilocularis in Central Italy

<p>Abstract</p> <p>Background</p> <p>Recent studies on geographic distribution of <it>Echinococcus multilocularis </it>in Europe show that it has a wider range than previously thought. It is unclear, however, if the wider distribution is due to its recent spreading or to a lack of previous data from the new areas. Italy, previously considered <it>E. multilocularis</it>-free, is now part of these new areas: infected foxes (the main definitive host of the tapeworm) have been observed in a Northern Alpine territory. Thus, more surveys need to be done in other Italian regions in order to monitor the spreading of <it>E. multilocularis</it>. The aim of the present study was to look for this parasite in 283 foxes collected in an Apennine area of Central Italy by different diagnostic methods.</p> <p>Results</p> <p>The foxes were heavily parasitized by 11 helminthic genera, but none of the animals was infected by <it>E. multilocularis </it>neither by <it>E. granulosus </it>(harboured adult worms or their DNA). Low specificity was observed in commercially available ELISA kits for the detection of <it>E. multilocularis </it>antigens in the faeces. Molecular diagnostics were sensitive and specific for the detection and identification of tapeworm eggs in faeces, but less sensitive, although specific, to adult tapeworms in the intestinal content.</p> <p>Conclusion</p> <p>Preliminarily, we can say that no <it>E. multilocularis </it>could be found in the study area. These data will enable us to follow temporal changes of the spatial distribution of the parasite in the study area of the Central Apennines. Due to its low specificity the ELISA kit for <it>E. multilocularis </it>coproantigens is not suitable for epidemiological surveys, whereas molecular diagnostics applied to faecal samples give useful results. Finally, absence of <it>E. granulosus </it>in foxes living in the endemic areas studied confirms the thought that this tapeworm prefers a different definitive host.</p

Subconjunctival Infection due to Dirofilaria Repens - Case Report

Summary The aim of the survey was to present the clinical course and surgical treatment of the first case of human ocular dirofilariosis on the territory of the city of Niš, in the southeast Serbia. Male patient, 57 years old, visited an ophthalmologist because of extreme swelling and redness of the eyelids of the right eye, scratches and pain in his right eye. On standard examination on biomicroscope, temporally 3 mm from the limbus, intrapalpebrally, a mobile parasite was observed in the subconjunctival space. Complete extraction of the living parasites, 13 cm long, was performed. A sample of the nematode based on morphological and morphometric characteristics was identified as Dirofilaria repens-like. The diagnosis was confirmed with molecular methods. For ocular dirofilariosis, surgical methods and complete extraction of the parasite are the only ways to achieve complete recovery

Human Babesiosis, Bolivia, 2013

To investigate human babesiosis in the Bolivian Chaco, in 2013 we tested blood samples from 271 healthy persons living in 2 rural communities in this region. Microscopy and PCR indicated that 3.3% of persons were positive for Babesia microti parasites (US lineage); seroprevalence was 45.7%. Appropriate screening should mitigate the risk for transfusion-associated babesiosis

Canine Dirofilaria Infections in Two Uninvestigated Areas of Serbia: Epidemiological and Genetic Aspects

In 2009 canine filarial infections were investigated in two northern areas of Serbia (Pancevo and Veliko Gradiste), applying morphometry, biochemical staining, and immunological kit to detect Dirofilaria immitis antigens, and two home-made ELISAs to detect antibodies to D. repens and D. immitis somatic/metabolic polyproteins. Moreover, molecular tools were applied to analyze the phylogenetic relationships of the isolates. The microfilariae detected in 21/122 dogs (17.2%) were identified as D. repens (n = 21) and D. immitis (n = 2). D. immitis antigens were found in another 13 animals with occult infection. All of the 15 heartworm-positive dogs also had antibodies to this parasite, which were detected in another 13 subjects, indicating an overall D. immitis seroprevalence rate of 22.9%. Serology for D. repens revealed evidence of antibodies in 42.6% of the dogs, but was negative for 4 microfilaremic dogs. As for the two different areas, the prevalence of microfilariae and/or D. immitis antigens, mainly due to D. repens microfilaremic animals, was not significantly higher in Veliko Gradiste (33.3%) than in Pancevo (22%). However, serology showed a different epidemiological picture. Heartworm infection occurred more often in both areas, and antibodies to dirofilarial nematodes were detected in 72.9% of dogs living in Pancevo, a rate higher than in those living in Veliko Gradiste (57.1%). No risk factors for infection were found, confirming the uselessness of prophylactic drugs against D. repens, and suggesting the presence in these areas of sunrise- or sunset-biting mosquitoes as important vectors. The results indicate the need for both appropriate entomological studies and further research on the intra-species variability shown by D. repens