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Background  Dirofilariosis is an emerging vector-borne parasitic disease in Europe. Monitoring of wild and domestic carnivores demonstrated circulation of  Dirofilaria  spp. in Romania in the past. For the implementation of control measures, knowledge on the native mosquito community responsible for  Dirofilaria  spp. transmission is required.    Methods  Mosquito samples originated from a longitudinal study previously performed in the Danube Delta Biosphere Reserve. Mosquito pools were screened for  Dirofilaria immitis  and  Dirofilaria repens . The samples comprised 240,572 female mosquito specimens collected every ten days between April and September in 2014 at four different trapping sites. In addition, blood samples of 36 randomly selected dogs were collected in 2016 in each of the four mosquito sampling sites. A duplex real-time assay was used to detect the presence of one or both  Dirofilaria  species for each sample. This assay targets the cytochrome  c  oxidase subunit 1 and the  16S  rRNA gene fragments to differentiate both parasites.    Results   Dirofilaria immitis  and  D. repens  were detected in mosquito pools at all four trapping sites. In the 2118 mosquito pools tested,  D. immitis  was identified for eight and  D. repens  for six of the 14 screened mosquito taxa, with a higher prevalence of  D. immitis  (4.53% of analysed pools) compared to  D. repens  (1.09%).  Dirofilaria  spp. were also identified in dogs from the same sampling sites with a prevalence of 30.56%. For both  Dirofilaria  species, the highest estimated infection rates (EIRs) were found in  Anopheles maculipennis  ( s.l. ) ( D. immitis : EIR = 0.206 per 100 specimens,  D. repens : EIR = 0.066 per 100 specimens). In contrast,  Coquillettidia richiardii  and  Anopheles hyrcanus  as the most frequent taxa had infection rates which were significantly lower:  Cq. richiardii  ( D. immitis : EIR = 0.021;  D. repens : EIR = 0.004);  An. hyrcanus  ( D. immitis : EIR = 0.028;  D. repens : EIR = 0.006). The number of positive pools per calendar week was positively correlated with the number of screened pools per calendar week, suggesting constant  Dirofilaria  spp. transmission during the observation period.    Conclusions  This study further confirms significant circulation of  Dirofilaria  spp. in eastern Europe, with high parasite prevalence in domestic canids and mosquitoes. Therefore, systematic monitoring studies are required to better understand the environmental risk factors for  Dirofilaria  transmission, allowing the implementation of effective surveillance and control measures.

Circulation of Dirofilaria immitis and Dirofilaria repens in the Danube Delta Biosphere Reserve, Romania