In 2006 and 2007, a total of 193 blood samples were collected from privately owned dogs housed predominantly outdoors, resident in four provinces in Vojvodina, Serbian Republic. Circulating microfilariae of Dirofilaria repens, Dirofilaria immitis and Acanthocheilonema reconditum were found in 49.2%, 7.2% and 2.1% of dogs, respectively. Two additional occult heartworm infections were revealed with an antigen test of 90 amicrofilaraemic dogs. Prevalence values were not influenced by sex and breed, but D. repens was significantly more prevalent in dogs > or =6 years old, and differences were observed between provinces. This is the first detailed survey on canine dirofilariasis in the continental part of the Balkans. Vojvodina is shown to have the northernmost limit of heartworm infection in the Balkan area and one of the zones in Europe with the highest prevalence of canine subcutaneous dirofilariasis. Chemoprophylaxis should be considered for resident dogs and dogs visiting this region.

In 2009 canine filarial infections were investigated in two northern areas of Serbia (Pančevo and Veliko Gradište), 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 Gradište (33.3%) than in Pančevo (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 Pančevo, a rate higher than in those living in Veliko Gradište (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.

BackgroundThe Northern part of Serbia is hyperendemic-endemic for canine dirofilarioses. Considering this fact, many human dirofilarial infections could be expected, however only about 30 cases in Serbia have been described until today. Aims of this survey were to assess the people reactivity to the antigens of D. repens and D. immitis and to identify risk factors for the contact exposure.MethodsInvestigation included sera taken from 297 people (179 women and 118 men) living in different areas of Serbia (Pančevo, Novi Sad, Zaječar, Leskovac, Vranje, Niš, Pirot). Sera were analysed by means of two indirect enzyme-linked immunosorbent (ELISA) home-designed that use as antigens adult somatic/metabolic polyproteins of D. repens (DR) and D. immitis (DI), respectively. The results were elaborated using the statistical method of descriptive and quantitative analysis.ResultsSignificant differences by area in the reactivity of human sera to dirofilarial antigens were not observed (p = 0.056). A high seroreactivity was demonstrated in people from the towns of northern Serbia (Pančevo = 27,1%; Novi Sad = 16,3%), as well as in people from Zaječar (eastern Serbia = 15,8%) and Vranje (southern Serbia = 15,1%). No differences were evidenced between people reactivity to polyproteins of the two dirofilarial species, nor differences related to the gender of examinees. Factor risks evidenced were: i) place of residence; ii) spending work time outdoors during the mosquito season; iii) spending time outdoors and nearby rivers, lakes, swamps or canals; unespectedly, iv) cat owning.ConclusionThe findings emerging from this investigation indicate that clinicians and public health authorities should pay greater attention to this zoonosis. Continuing education and training of physicians will greatly contribute to the knowledge of the actual impact of filarial worms on animal and public health, and allow for the planning of suitable measures to prevent the infections.

Human dirofilariosis is a relatively rare infection caused by filarial worms of the genus Dirofilaria. We herein report the first case of human subcutaneous dirofilariosis in the southeastern part of Serbia. A complete alive nematode was removed from a nodule in the periorbital region of a 50-year-old woman. The nematode was morphologically identified as a D. repens-like immature female. The diagnosis was confirmed with molecular methods. The patient was probably infected in the South-East Serbia as she had not travelled abroad, nor in other parts of Serbia such as Vojvodina, recently identified as a hyperendemic area for D. repens infection of dogs.

Infection with parasites from the Trichinella genus occurs in many vertebrates but disease only occurs in humans (trichinellosis). Humans are infected after the consumption of raw or undercooked meat from infected wild or domestic animals (usually swine or horses). Using the monoclonal antibody (mAb) 7C2C5, specific for an epitope unique to the muscle larvae of the genus Trichinella, we have developed a competitive enzyme-linked immunosorbent assay (c-ELISA) that enables the rapid detection of Trichinella-specific antibodies in sera originating from two different host species (human, swine) infected with either Trichinella spiralis or Trichinella britovi. This novel c-ELISA exhibited 100% specificity and sensitivity, as confirmed by a Western blot test. The assay is easy to use (one incubation step), and the time required for the procedure (45 min) is shorter than in any other ELISA format. This test could be useful for both the detection and surveillance of Trichinella infections.