Human infection by Dirofilaria repens in Serbia has been increasing steadily. The first case was reported in 1971, presented in the form of a single subcutaneous nodule on the back of a young boy. As established by a literature search, eight additional cases were reported until mid-2001. The most frequent site of infection was subcutaneous tissue, with the exception of two cases, in which parasites were found in subconjunctiva and epididymis. Our study, conducted from 2001 to 2008, encompasses 19 new cases. Most of them (63.1%) presented as ocular or periocular infections, in which the parasite was typically found under the conjunctiva. In other cases a parasitic nodule was localized in the temporal region of the head, epididymis, testicle, abdomen, breast or arm. The diagnosis was made by morphological and histological analysis of the extracted intact worms and parasite sections from the tissue. Morphology of the filarial worms was well preserved in more than half of the cases (12/19) and there was never more than one parasite found inside the lesions. Adult worms and immature nematodes were observed in nine and seven cases, respectively. Furthermore, in two cases microfilariae were discovered inside the pseudocoelom, sections of the female reproductive tubes filled with clearly visible larval stages. Dirofilaria repens infection was diagnosed by its morphological features (17/19) or by performing polymerase chain reactions (PCR) using paraffin-embedded tissues (2/19) in the cases where the morphology was insufficient for identification and the parasites had been determined initially as Dirofilaria spp. The amplified 246 bp PCR product showed that the worms were D. repens.
Cystic echinococcosis (CE) is a widespread and severe zoonotic disease caused by infection with the larval stage of the eucestode Echinococcus granulosus sensu lato. The polymorphism exhibited by nuclear and mitochondrial markers conventionally used for the genotyping of different parasite species and strains does not reach the level necessary for the identification of genetic variants linked to restricted geographical areas. EmsB is a tandemly repeated multilocus microsatellite that proved its usefulness for the study of genetic polymorphisms within the species E. multilocularis, the causative agent of alveolar echinococcosis. In the present study, EmsB was used to characterize E. granulosus sensu lato samples collected from different host species (sheep, cattle, dromedaries, dogs, and human patients) originating from six different countries (Algeria, Mauritania, Romania, Serbia, Brazil, and the People's Republic of China). The conventional mitochondrial cox1 and nad1 markers identified genotypes G1, G3, G5, G6, and G7, which are clustered into three groups corresponding to the species E. granulosus sensu stricto, E. ortleppi, and E. canadensis. With the same samples, EmsB provided a higher degree of genetic discrimination and identified variations that correlated with the relatively small-scale geographic origins of the samples. In addition, one of the Brazilian single hydatid cysts presented a hybrid genotypic profile that suggested genetic exchanges between E. granulosus sensu stricto and E. ortleppi. In summary, the EmsB microsatellite exhibits an interesting potential for the elaboration of a detailed map of the distribution of genetic variants and therefore for the determination and tracking of the source of CE.
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