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Background   Dirofilaria immitis  and  Dirofilaria repens  are transmitted by bloodsucking culicid mosquitoes belonging to  Culex, Aedes, Ochlerotatus ,  Anopheles  and  Mansonia  genera.  The detection of filarioids in mosquitoes for assessing distribution of vectors and/or of pathogens in a given area (also known as “xenomonitoring”), when based on individual dissection of wild-caught female mosquitoes is time consuming and hardly applicable in large epidemiological surveys.  Our study aimed to evaluate the recently developed duplex real-time PCR for screening large number of culicids and to assess their positivity for  D. immitis  and  D. repens  in an area where both species are endemic.    Methods  A duplex real-time PCR was used to detect and differentiate  D. immitis  and  D. repens  in mosquitoes collected in six provinces of the Veneto region using 43 carbon dioxide-baited traps under the frame of an entomological surveillance program to monitor the vectors of West Nile disease. From early May till October 2010, unfed female mosquitoes (n = 40,892) were captured in 20 selected sites.    Results  Mosquitoes identified as  Culex pipiens ,  Ochlerotatus caspius ,  Aedes vexans  and  Culex modestus  were grouped into 995 pools according to species, day and site of collection (from minimum of 1 to maximum of 57). Out of 955 pools, 23 (2.41 %) scored positive for  Dirofilaria  spp. of which, 21 (2.2 %) for  D. immitis  and two (0.21 %) for  D. repens . An overall Estimated Rate of Infection (ERI) of 0.06 % was recorded, being higher in  Och. caspius  and  Ae. vexans  (i.e., 0.18 % and 0.14 %, respectively). At least one mosquito pool was positive for  Dirofilaria  spp. in each province with the highest ERI recorded in Vicenza and Padova provinces (i.e., 0.42% and 0.16 %, respectively). Mosquitoes collected in all provinces were positive for  D. immitis  whereas, only two (i.e., Padova and Rovigo) provinces scored positive for  D. repens . All mosquito species, except for  Cx. modestus , were positive for  D. immitis,  whereas  D. repens  was only found in  Cx. pipiens .    Conclusions  The results suggest that both  Dirofilaria  species are endemic and may occur in sympatry in the examined area. The molecular approach herein used represents a powerful tool for surveillance programs of  D. immitis  and  D. repens  in the culicid vectors towards a better understanding of the epidemiology of the infections they cause and their seasonal transmission patterns.

parasites and vectors

Molecular xenomonitoring of Dirofilaria immitis and Dirofilaria repens in mosquitoes from north-eastern Italy by real-time PCR coupled with melting curve analysis