Identification of vectors is of prime importance in the field of medical entomology for both operational and research purposes. An external quality assessment of mosquito identification capacities was carried out within the MediLabSecure Network, which is composed of laboratories located in 19 countries close to the European Union around the Mediterranean and Black seas. A set of blind samples consisting of 7 or 8 adult mosquitoes and 4 larvae was given to each participant laboratory. In all, 138 adult mosquitoes and 76 larvae of different species were distributed for genus and species identification. All identifications were exclusively morphology based. Overall, 81% of identifications were correct at the genus level, 64% at the species level. The results were highly varied among the 19 participating laboratories. The levels of correct identifications were: 100% (three laboratories), 90–95% (four laboratories), 50–75% (six laboratories) and < 50% (six laboratories). This evaluation showed the need to maintain efforts in capacity building and quality control in the field of medical entomology and, more specifically, in the morphological identification of the Culicidae.

The aim of this study was to examine the seroprevalence of Leptospira spp. in dogs and red foxes in the entity of the Republic of Srpska, Bosnia and Herzegovina, after heavy rainfall and floods in 2014 and for the two years thereafter. The seroepidemiological study involved testing serum samples from dogs (n = 98) and foxes (n = 112) using MAT (microscopic agglutination test). Antibodies to at least one Leptospira spp. serovar were found in 52.04% of the tested dogs. The dog seroprevalence in 2014 (81.25%) was significantly higher than in 2015 (51.42% p <0.0001) and 2016 (22.5% p<0.05). The highest seroprevalences were for serovars Australis (76.47%), Bratislava (70.58%), Sejroe (66.67%) and Autumnalis (45.09%). Antibodies to at least one Leptospira spp. serovar were detected in 34.82% of the examined red foxes. In 2015, the fox seroprevalence was significantly higher (52.94%) than in 2016 (6.82%) (p <0.0001). The highest seroprevalences were for serovars Sejroe (64.10%), Bratislava (48.72%), Australis (43.59%) and Bataviae (25.64%). The high seroprevalence of Leptospira spp. in dogs and foxes determined during this study indicates the importance of these carnivores in maintaining leptospirosis in the study area, and the potential risk of infection for humans and other animal species that come into contact with these canids. The results obtained indicate that heavy rainfall and intense floods can result in increased Leptospira spp. infection in these canids.

This study compared serological and molecular approaches for detecting Leptospira spp. using paired blood and urine samples from 124 asymptomatic stray dogs. Blood sera were screened via the microscopic agglutination test (MAT) for ten serovars. Urine samples were analyzed using four PCR assays: a commercial quantitative PCR (qPCR), a lipL32 qPCR, a conventional rrs PCR, and a multiplex PCR (mPCR). Serological exposure was detected in 26 (20.97%) dogs, while commercial qPCR, rrs PCR, lipL32 qPCR, and mPCR assays yielded positive results in 31 (25.00%), 34 (27.42%), 25 (20.16%), and 23 (18.55%) dogs, respectively. Compared with commercial qPCR, the lipL32 qPCR, rrs PCR, and mPCR demonstrated Positive Percentage Agreement (PPA) values of 80.65%, 87.10%, and 67.74%, and Negative Percentage Agreement (NPA) values of 100.00%, 92.47%, and 97.85%. Nineteen urine samples were positive across all four PCR assays. Overall, 42.74% (53/124) of dogs had at least one positive serological or molecular finding, including nine additional conventional PCR-positive findings without reliable sequence confirmation. Sequencing and phylogenetic analysis of 24 samples identified L. interrogans and L. borgpetersenii, representing the first molecular characterization of Leptospira in Bosnia and Herzegovina. The findings demonstrate the complementary value of serological and molecular approaches for Leptospira surveillance while emphasizing the need for cautious interpretation of unconfirmed molecular findings.