Culicoides (Diptera: Ceratopogonidae) is a genus of small biting midges (also known as “no-see ums”) that currently includes 1368 described species. They are proven or suspected vectors for important pathogens affecting animals such as bluetongue virus (BTV) and Schmallenberg virus (SBV). Currently little information is available on the species of Culicoides present in Serbia. Thus, the aim of this study was to examine species diversity, host preference and the presence of BTV and SBV RNA in Culicoides from the Stara Planina Nature Park in south-eastern Serbia. In total 19,887 individual Culicoides were collected during three nights of trapping at two farm sites and pooled into six groups (Obsoletus group, Pulicaris group, “Others” group and further each group according to the blood-feeding status to freshly engorged and non-engorged). Species identification was done on subsamples of 592 individual Culicoides specimens by morphological and molecular methods (MALDI-TOF mass spectrometry and PCR/sequencing). At least 22 Culicoides species were detected. Four animal species (cow, sheep, goat and common blackbird) as well as humans were identified as hosts of Culicoides biting midges. The screening of 8291 Culicoides specimens in 99 pools for the presence of BTV and SBV RNA by reverse-transcription quantitative PCR were negative. The biodiversity of Culicoides species in the natural reserve Stara Planina was high with at least 22 species present. The presence of C. imicola Kieffer was not recorded in this area. Culicoides showed opportunistic feeding behaviour as determined by host preference. The absence of SBV and BTV viral RNA correlates with the absence of clinical disease in the field during the time of sampling. These data are the direct outcome of a training programme within the Institutional Partnership Project “AMSAR: Arbovirus monitoring, research and surveillance-capacity building on mosquitoes and biting midges” funded by the programme SCOPES of the Swiss National Science Foundation.

The prime significance of species belonging to the genus Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) is their ability to transmit viruses such as bluetongue virus (BTV) to wild and domestic ruminants. Prior to 1998, BTV was considered exotic in Europe, but according to recent history of its outbreaks, it has become endemic in southern and eastern European countries circulating beyond its expected historical limits, into the Balkan region. The wind-borne long-distance dispersal of Culicoides spp. over water bodies and local spreading between farms emphasize the necessity of filling in the information gaps regarding vector species distribution. In most Balkan countries, data on Culicoides fauna and species distribution are lacking, or information is old and scarce. During this study, 8586 specimens belonging to 41 species were collected. We present the first faunistic data on Culicoides species in the former Yugoslav Republic of Macedonia (FYROM), Kosovo, Montenegro and Serbia. For other countries (Bosnia and Herzegovina, Bulgaria and Croatia), all historical records were compiled for the first time and then expanded with our findings to various extents. In all countries, confirmed or suspected BTV vector species belonging to the subgenera Avaritia and Culicoides were collected. The total number of species sampled during our field collections was 20 in Bosnia and Herzegovina (15 new records), 10 in Bulgaria (2 new records), 10 in Croatia (5 new records), 13 in FYROM, 9 in Kosovo, 15 in Montenegro, and 28 in Serbia. Of these, 14 species were registered for the first time in this part of the Balkans. This paper provides the first data about Culicoides fauna in FYROM, Kosovo, Montenegro and Serbia, as well as new records and an update on the checklists for Bosnia and Herzegovina, Bulgaria and Croatia. These findings provide preliminary insights into the routes of BTV introduction and spreading within the Balkans, and present a valuable contribution to further research related to Culicoides-borne diseases in Europe.

Aedes albopictus is currently the most widespread invasive mosquito species in the world. It has paramount medical importance since females are efficient vectors of important viruses affecting humans. The development of alternative control strategies to complement control measures has become an imperative and involves the Sterile Insect Technique (SIT). Research to improve the productivity of mass-rearing, as well as the quality of mass-reared males is of essential importance for the success of SIT. This study compared the influence of three differently sized cages for Ae. albopictus mass-rearing on wing length, adult survival and egg production during 20 generations of colonization. Plexiglas cages of 40x40x40 cm (C1), 100 × 20 × 100 cm (C2) and 100 × 65 × 100 cm (C3) were loaded with equal adult density, and sex ratio of 1:1. An open source image processing and analysis programme (ImageJ) was used for the wing measurement and egg counting. In all tested cages, we identified two periods separated by the generation showing the minimum value of each considered parameter (wing length, adult survival and egg production). The wing length and adult survival passed through the phases of initial decrease to about intermediate colonization time, and increased afterwards. Fecundity was steady during the first period and increased in the second one. Cage C1 demonstrated not only the best values for all parameters but also the smallest decrease in the initial phase. Recovering of the caged mosquitoes in the second half of the study was higher in cages C1 and C2, than in C3. C1 provided the least negative selection pressure on wing length, adult survival and egg production for reared Ae. albopictus. Anyhow, since maximising mosquito density by exploiting the minimum space is a priority in mosquito mass-rearing, C2 might be a better choice for better fitting the space of mass-rearing rooms.

This is the third in a planned series of data papers presenting modelled vector distributions produced during the ECDC and EFSA funded VectorNet project. The data package presented here includes those Culicoides vectors species first modelled in 2015 as part of the VectorNet gap analysis work namely C. imicola, C. obsoletus, C. scoticus, C. dewulfi, C. chiopterus, C. pulicaris, C. lupicaris, C. punctatus, and C. newsteadi. The known distributions of these species within the Project area (Europe, the Mediterranean Basin, North Africa, and Eurasia) are currently incomplete to a greater or lesser degree. The models are designed to fill the gaps with predicted distributions, to provide a) first indication of vector species distributions across the project geographical extent, and b) assistance in targeting surveys to collect distribution data for those areas with no field validated information. The models are based on input data from light trap surveillance of adult Culicoides across continental Europe and surrounding regions (71.8°N –33.5°S, – 11.2°W – 62°E), concentrated in Western countries, supplemented by transect samples in eastern and northern Europe. Data from central EU are relatively sparse. Funding statement: This work was carried out with support from the VectorNet framework contract OC/EFSA/AHAW/2013/02-FWC1 funded by the European Centre for Disease prevention and Control (ECDC) and the European Food Safety Agency (EFSA) and the PALE-Blu H2020 Project ID: 727393.

Culex pipiens is a widespread mosquito species with high ecological plasticity that thrives in urban, peri-urban and rural aquatic habitats. It is a major vector of West Nile virus (WNV), contributing to virus transmission among bird reservoirs and serving as a bridge vectorfor transmission to humans and mammals, which can result in neuroinvasive disease and fatalities. Controlling its populations reduces biting nuisance and associated economic and health burdens, making vector management essential for effective public health protection. Available methods to control this species are limited and require significant improvement because conventional strategies are often short-term, non-specific and ecologically problematic. The present study evaluated the efficacy of cyromazine granules on the Biodac carrier in laboratory, semi-field (in barrels) and field experiments (in canals). Content of cyromazine was 0.5 or 2%. Two formulations were tested: granules coated with stearate and uncoated granules. The highest efficacy was demonstrated by application of 2% cyromazine, both coated and uncoated, compared to the cyromazine with 0.5% active substance. Cyromazine showed high efficacy in the control of Cx. pipiens ranging from 85.8% to 100% in the laboratory, 68.1% to 100% in the semi-field and 48.1% to 98.8% in the field conditions (depending on the formulation applied), enabling long-lasting suppression of juvenile stages. In the laboratory, 53 days post-treatment, the residues were still present in the water. In the field experiment (in canals) the population reduction was recorded up to the 56th day post-treatment.