Dirofilaria sp. and Blood Meal Analysis in Mosquitoes Collected in Vojvodina and Macva, and the First Report of Setaria tundra (Issaitshikoff & Rajewskaya, 1928) in Serbia.

Dirofilaria immitis and D. repens are the two most widespread and important species of mosquito-borne nematodes, posing a significant threat to veterinary health and particularly affecting canines and felines. While D. immitis causes cardiopulmonary dirofilariasis, D. repens causes subcutaneous infections in dogs and other carnivores. Despite the extensive knowledge on these parasites, little is known about their natural vectors in Serbia. The parasite Setaria tundra, known to infect deer, has not yet been detected in Serbia but has been documented in neighboring countries. Thus, the aim of this study was to (i) further map out Dirofilaria sp. hotspots in the Vojvodina Province and detect S. tundra for the first time, (ii) detect positive mosquito species that can provide insights into how the nematodes spread in Serbia, and (iii) analyze the blood-fed female mosquitoes of species found to be infected, in order to identify the potential source of parasite infection. A total of 2902 female mosquitoes were collected across 73 locations during 2021 and 2022. Molecular biology methods, based on conventional PCR, were used to analyze non-blood-fed (2521 specimens) and blood-fed (381 specimens) female mosquitos, in order to detect filarial nematode presence and identify blood-meal sources, respectively. When the parasite genome was detected, the amplicon (cox1 gene, 650 bp fragment) was sent for Sanger sequencing, further confirming the presence of nematodes and species assignation. D. immitis was detected in three Culex pipiens mosquitoes collected in Zrenjanin (August 2021) and Glogonj and Svetozar Miletic (both in July 2021). Additionally, Setaria tundra was detected in Aedes vexans collected in Idos (mid-August 2021) and Aedes caspius, which was collected in Mali Idos (end of July 2021). This work identifies two new locations where D. immitis occurs in Vojvodina, and is the first report of S. tundra in Serbian territory. Blood-meal analysis provided insights into the preferences of mosquitoes that were positive for Dirofilaria sp. and S. tundra.

The Introduction and Establishment of Four Invasive Insect Species in Serbia.

Urban areas are often populated by specific species of insects, some colorful and appealing, such as ladybugs and butterflies, and others irritating as nuisance bitters or as vectors of pathogens of public health importance. Mosquitoes in urban areas often utilize habitats adjacent to human residences, while phytophagous insect species such as stink bugs often colonize ornamental plants and utilize human-made structures including houses as overwintering shelters. This article discusses the early detection and the current distribution of two invasive mosquito species, Aedes albopictus Skuse 1894 and Ae. japonicus (Theobald 1901), in Serbia, introduced in 2009 and 2018, respectively. From the first findings until today, regular monitoring has been carried out and the establishment of both species in the newly invaded areas has been confirmed. Both species can become nuisance species, especially at high population densities, but more importantly, they are capable of transmitting a wide variety of arboviruses of public health importance. This article also discusses two invasive stink bug species Halyomorpha halys Stål 1855 and Nezara viridula Linnaeus 1758, introduced in Serbia in 2015 and 2008, respectively. These two stink bug species have also been monitored, and the establishment of their populations in the country has been confirmed. Both species have caused damage to a wide range of crops and ornamental plants and sometimes become nuisance pests in urban areas.

Two invasions at once: update on the introduction of the invasive species Aedes aegypti and Aedes albopictus in Cyprus - a call for action in Europe.

Aedes aegypti, the yellow fever mosquito and Aedes albopictus, the tiger mosquito, continue to expand their geographical distribution, reshaping the European epidemiological risks for mosquito-borne diseases. The reintroduction of Aedes aegypti near the airport and port in Larnaka and the detection of Aedes albopictus near the marina and old port of the Limassol area in Cyprus are reported herein. The measures taken to investigate these events included (i) communication to health authorities, (ii) expert on-site visits and verification of findings, (iii) enhanced active surveillance, and (iv) development of an Emergency Action Plan followed by a Contingency Plan. These emergency action plans were developed to delimitate the infested areas and to prevent the spreading of the mosquito populations into new areas. The general principles are presented along with their rationale to serve as guidelines for other geographical regions targeting suppression/eradication with a sterile insect technique component. In parallel, this manuscript serves as a call for action at the European level to impede the further spread of these species and support the activities being undertaken in Cyprus to combat the incursions of Aedes invasive species.

Title: Deux invasions à la fois : le point sur l'introduction des espèces envahissantes Aedes aegypti et Aedes albopictus à Chypre ­ un appel à l'action en Europe. Abstract: Aedes aegypti, le moustique de la fièvre jaune, et Aedes albopictus, le moustique tigre, continuent à étendre leurs aires de distribution, redessinant les risques épidémiologiques des maladies transmises par les moustiques en Europe. La réintroduction d'Aedes aegypti près de l'aéroport et du port de Larnaka, et la détection d'Aedes albopictus près de la marina et du vieux port de la zone de Limassol à Chypre sont rapportées dans cette étude. Les mesures prises pour investiguer ces évènements ont inclus : (i) la communication avec les autorités sanitaires, (ii) des visites d'experts sur site et la vérification des résultats, (iii) une surveillance active augmentée et (iv) le développement d'un Plan d'Actions d'Urgence suivi d'un Plan de Contingence. Des plans d'action d'urgence ont été développés pour délimiter les zones infestées et prévenir l'expansion des populations de moustiques à d'autres zones. Les principes généraux sont présentés, en même temps que leur justification, pour guider d'autres zones géographiques visant la suppression / éradication avec une composante de technique de l'insecte stérile. En parallèle, cet article sert d'appel, au niveau européen, à prévenir la diffusion toujours plus importante de ces espèces et pour appuyer les activités à mener à Chypre pour combattre les incursions des espèces invasives d'Aedes.

Bio-efficacy of permethrin/tetramethrin and lambda-cyhalothrin treatments in habitats of hard ticks (Acari, Ixodidae) populations with confirmed Borrelia spp. infection.

The aim of the study was to evaluate the bio-efficacy of two different acaricides against mobile stages of hard ticks Ixodes ricinus, Dermacentor marginatus, and Haemaphysalis punctata in their natural habitats. The study was conducted during 2020 and 2021 at localities populated by I. ricinus as the predominant species, at which the presence of Borrelia afzelii, Borrelia garinii, and Borrelia lusitaniae was confirmed. During the first investigation year, a combination of two pyrethroids, permethrin, and tetramethrin, with an insecticide synergist piperonyl butoxide (trade name: Perme Plus®) was tested. At the first evaluation, 24 h after the treatment with Perme Plus®, the efficacy expressed as a reduction rate of the population density was within the interval of satisfying performance (70-90%) at all localities, while the highest efficacy (97.8%) was recorded on the 14th post-treatment day. In the second investigation year, the formulation based on lambda-cyhalothrin (trade name: Icon® 10CS) was used. On the first post-treatment evaluation day, satisfying effects were also demonstrated. The highest recorded efficacy rate of lambda-cyhalothrin (94.7%) was recorded on the 14th post-treatment day. Both tested acaricides manifested satisfying initial acaricidal effects against mobile stages of ticks and provided long-term effects. Comparison of the regression trend lines of population reduction revealed that satisfying effects of treatment with Perme Plus® lasted until the 17th post-treatment day, while in the case of Icon® 10CS, the residual effects were significantly prolonged (30 days).

Assessment of expertise in morphological identification of mosquito species (Diptera, Culicidae) using photomicrographs.

Accurate identification of insect species is an indispensable and challenging requirement for every entomologist, particularly if the species is involved in disease outbreaks. The European MediLabSecure project designed an identification (ID) exercise available to any willing participant with the aim of assessing and improving knowledge in mosquito taxonomy. The exercise was based on high-definition photomicrographs of mosquitoes (26 adult females and 12 larvae) collected from the western Palaearctic. Sixty-five responses from Europe, North Africa and the Middle East were usable. The study demonstrated that the responders were better at identifying females (82% correct responses) than larvae (63%). When the responders reported that they were sure of the accuracy of their ID, the success rate of ID increased (92% for females and 88% for larvae). The top three tools used for ID were MosKeyTool (72% of responders), the ID key following Becker et al. [2010. Mosquitoes and their control, 2nd edn. Berlin: Springer] (38%), and the CD-ROM of Schaffner et al. [2001. Les moustiques d'Europe: logiciel d'identification et d'enseignement - The mosquitoes of Europe: an identification and training programme. Montpellier: IRD; EID] (32%), while other tools were used by less than 10% of responders. Responders reporting the identification of mosquitoes using the MosKeyTool were significantly better (80% correct responses) than non-MosKeyTool users (69%). Most responders (63%) used more than one ID tool. The feedback from responders in this study was positive, with the exercise being perceived as halfway between educational training and a fun quiz. It raised the importance of further expanding training in mosquito ID for better preparedness of mosquito surveillance and control programmes.

Title: Évaluation de l'expertise en identification morphologique des espèces de moustiques (Diptera, Culicidae) à l'aide de photomicrographies. Abstract: L'identification précise des espèces d'insectes est une exigence indispensable et difficile pour tout entomologiste, en particulier si l'espèce est impliquée dans des épidémies. Le projet européen MediLabSecure a conçu un exercice d'identification (ID) accessible à tout participant volontaire dans le but d'évaluer et d'améliorer les connaissances en taxonomie des moustiques. L'exercice était basé sur des photomicrographies haute définition de moustiques (26 femelles adultes et 12 larves) prélevées dans le Paléarctique occidental. Soixante-cinq réponses d'Europe, d'Afrique du Nord et du Moyen-Orient ont été utilisables. L'étude a démontré que les répondants étaient meilleurs pour identifier les femelles (82 % de réponses correctes) que les larves (63 %). Lorsque les répondants ont déclaré être sûrs de l'exactitude de leur ID, le taux de réussite de l'identification était meilleur (92 % pour les femelles et 88 % pour les larves). Les trois principaux outils utilisés pour les ID étaient MosKeyTool (72 % des répondants), la clé d'identification du livre de Becker et al. (38%) et le CD-ROM de Schaffner et al. (32 %), tandis que d'autres outils étaient utilisés par moins de 10 % des répondants. Les répondants déclarant identifier des moustiques à l'aide de MosKeyTool étaient significativement meilleurs (80 % de réponses correctes) que les non-utilisateurs de MosKeyTool (69 %). La plupart des répondants (63 %) ont utilisé plus d'un outil d'identification. Les commentaires des répondants de cette étude ont été positifs, l'exercice étant perçu comme à mi-chemin entre une formation pédagogique et un quiz amusant. Il a souligné l'importance d'étendre la formation complémentaire à l'identification des moustiques pour une meilleure préparation des programmes de surveillance et de contrôle des moustiques.

A Mark-Release-Recapture Study to Estimate Field Performance of Imported Radio-Sterilized Male Aedes albopictus in Albania.

The pathogen transmitting Aedes albopictus mosquito is spreading rapidly in Europe, putting millions of humans and animals at risk. This species is well-established in Albania since its first detection in 1979. The sterile insect technique (SIT) is increasingly gaining momentum worldwide as a component of area-wide-integrated pest management. However, estimating how the sterile males will perform in the field and the size of target populations is crucial for better decision-making, designing and elaborating appropriate SIT pilot trials, and subsequent large-scale release strategies. A mark-release-recapture (MRR) experiment was carried out in Albania within a highly urbanized area in the city of Tirana. The radio-sterilized adults of Ae. albopictus Albania strain males were transported by plane from Centro Agricoltura Ambiente (CAA) mass-production facility (Bologna, Italy), where they were reared. In Albania, sterile males were sugar-fed, marked with fluorescent powder, and released. The aim of this study was to estimate, under field conditions, their dispersal capacity, probability of daily survival and competitiveness, and the size of the target population. In addition, two adult mosquito collection methods were also evaluated: BG-Sentinel traps baited with BG-Lure and CO2, (BGS) versus human landing catch (HLC). The overall recapture rates did not differ significantly between the two methods (2.36% and 1.57% of the total male released were recaptured respectively by BGS and HLC), suggesting a similar trapping efficiency under these conditions. Sterile males traveled a mean distance of 93.85 ± 42.58 m and dispersed up to 258 m. Moreover, they were observed living in the field up to 15 days after release with an average life expectancy of 4.26 ± 0.80 days. Whether mosquitoes were marked with green, blue, yellow, or pink, released at 3.00 p.m. or 6.00 p.m., there was no significant difference in the recapture, dispersal, and survival rates in the field. The Fried competitiveness index was estimated at 0.28. This mark-release-recapture study provided important data for better decision-making and planning before moving to pilot SIT trials in Albania. Moreover, it also showed that both BG-traps and HLC were successful in monitoring adult mosquitoes and provided similar estimations of the main entomological parameters needed.

Geographic distribution of the V1016G knockdown resistance mutation in Aedes albopictus: a warning bell for Europe.

BACKGROUND: Colonization of large part of Europe by the Asian tiger mosquito Aedes albopictus is causing autochthonous transmission of chikungunya and dengue exotic arboviruses. While pyrethroids are recommended only to reduce/limit transmission, they are widely implemented to reduce biting nuisance and to control agricultural pests, increasing the risk of insurgence of resistance mechanisms. Worryingly, pyrethroid resistance (with mortality < 70%) was recently reported in Ae. albopictus populations from Italy and Spain and associated with the V1016G point mutation in the voltage-sensitive sodium channel gene conferring knockdown resistance (kdr). Genotyping pyrethroid resistance-associated kdr mutations in field mosquito samples represents a powerful approach to detect early signs of resistance without the need for carrying out phenotypic bioassays which require availability of live mosquitoes, dedicated facilities and appropriate expertise. METHODS: Here we report results on the PCR-genotyping of the V1016G mutation in 2530 Ae. albopictus specimens from 69 sampling sites in 19 European countries. RESULTS: The mutation was identified in 12 sites from nine countries (with allele frequencies ranging from 1 to 8%), mostly distributed in two geographical clusters. The western cluster includes Mediterranean coastal sites from Italy, France and Malta as well as single sites from both Spain and Switzerland. The eastern cluster includes sites on both sides of the Black Sea in Bulgaria, Turkey and Georgia as well as one site from Romania. These results are consistent with genomic data showing high connectivity and close genetic relationship among West European populations and a major barrier to gene flow between West European and Balkan populations. CONCLUSIONS: The results of this first effort to map kdr mutations in Ae. albopictus on a continental scale show a widespread presence of the V1016G allele in Europe, although at lower frequencies than those previously reported from Italy. This represents a wake-up call for mosquito surveillance programs in Europe to include PCR-genotyping of pyrethroid resistance alleles, as well as phenotypic resistance assessments, in their routine activities.

A dynamically structured matrix population model for insect life histories observed under variable environmental conditions.

Various environmental drivers influence life processes of insect vectors that transmit human disease. Life histories observed under experimental conditions can reveal such complex links; however, designing informative experiments for insects is challenging. Furthermore, inferences obtained under controlled conditions often extrapolate poorly to field conditions. Here, we introduce a pseudo-stage-structured population dynamics model to describe insect development as a renewal process with variable rates. The model permits representing realistic life stage durations under constant and variable environmental conditions. Using the model, we demonstrate how random environmental variations result in fluctuating development rates and affect stage duration. We apply the model to infer environmental dependencies from the life history observations of two common disease vectors, the southern (Culex quinquefasciatus) and northern (Culex pipiens) house mosquito. We identify photoperiod, in addition to temperature, as pivotal in regulating larva stage duration, and find that carefully timed life history observations under semi-field conditions accurately predict insect development throughout the year. The approach we describe augments existing methods of life table design and analysis, and contributes to the development of large-scale climate- and environment-driven population dynamics models for important disease vectors.

Optimization of Aedes albopictus (Diptera: Culicidae) Mass Rearing through Cost-Effective Larval Feeding.

Aedes (Stegomyia) albopictus (Skuse, 1895) is an invasive important medical and veterinary pest species. The sterile insect technique (SIT) involves the mass rearing of males, and their sterilization and release into the habitat to compete with wild males. Our research objective was to compare the effectiveness of three larval diet recipes (IAEA-BY, BCWPRL, and MIX-14) in the laboratory rearing of Ae. albopictus males to evaluate the available economical feeding alternatives. The separation of sexes was done in the pupal stage by sieving. Reared males were tested for flight capacity and longevity. The application of the BCWPRL diet resulted in a higher portion of sieved male pupae than females, but the development of males was the slowest, and the number of obtained males (pupae and adults) was lower compared to the other two diets. The adult mean survival time was the highest in males fed with MIX-14 and the lowest in males fed with IAEA-BY. Males fed by IAEA-BY also demonstrated higher initial mortality in the adult stage. The diets BCWPRL and MIX-14 are economically more convenient than IAEA-BY (2.28 and 5.30 times cheaper, respectively). The cheapest diet, MIX-14, might represent a candidate for replacing the effective but still expensive IAEA-BY larval diet, providing lower costs of sterile male production.

AIMSurv: First pan-European harmonized surveillance of Aedes invasive mosquito species of relevance for human vector-borne diseases.

Human and animal vector-borne diseases, particularly mosquito-borne diseases, are emerging or re-emerging worldwide. Six Aedes invasive mosquito (AIM) species were introduced to Europe since the 1970s: Aedes aegypti, Ae. albopictus, Ae. japonicus, Ae. koreicus, Ae. atropalpus and Ae. triseriatus. Here, we report the results of AIMSurv2020, the first pan-European surveillance effort for AIMs. Implemented by 42 volunteer teams from 24 countries. And presented in the form of a dataset named "AIMSurv Aedes Invasive Mosquito species harmonized surveillance in Europe. AIM-COST Action. Project ID: CA17108". AIMSurv2020 harmonizes field surveillance methodologies for sampling different AIMs life stages, frequency and minimum length of sampling period, and data reporting. Data include minimum requirements for sample types and recommended requirements for those teams with more resources. Data are published as a Darwin Core archive in the Global Biodiversity Information Facility- Spain, comprising a core file with 19,130 records (EventID) and an occurrences file with 19,743 records (OccurrenceID). AIM species recorded in AIMSurv2020 were Ae. albopictus, Ae. japonicus and Ae. koreicus, as well as native mosquito species.

Intensive West Nile Virus Circulation in Serbia in 2018-Results of Integrated Surveillance Program.

The results of the Serbian national integrated West Nile virus (WNV) surveillance program conducted in 2018 and funded by the Serbian Veterinary Directorate are presented. The WNV surveillance program encompassed the entire territory of Serbia and was conducted by the veterinary service in collaboration with entomologists and ornithologists. The objective of the program was early detection of WNV circulation in the environment and timely reporting to the public health service and local authorities to increase clinical and mosquito control preparedness. The program was based on the detection of WNV presence in wild birds (natural hosts) and mosquitoes (virus vectors) and on serological testing of sentinel horses (WNV-specific IgM antibodies). The season 2018 was confirmed to be the season of the most intensive WNV circulation with the highest number and severity of human cases in Serbia ever reported. The most intense WNV circulation was observed in the northern and central parts of Serbia including Vojvodina Province, the Belgrade City area, and surrounding districts, where most positive samples were detected among sentinel animals, mosquitoes and wild birds. The majority of human cases were preceded by the detection of WNV circulation during the surveillance. The WNV surveillance program in 2018 showed satisfactory results in the capacity to indicate the spatial distribution of the risk for humans and sensitivity to early detection of WNV circulation in the environment.

Rapid spread and population genetics of Aedes japonicus japonicus (Diptera: Culicidae) in southeastern Europe (Croatia, Bosnia and Herzegovina, Serbia).

The Asian bush mosquito, Aedes japonicus japonicus (Theobald, 1901), a potential vector of several pathogens, has recently established in North America and Central Europe. In 2013, it was found on the Slovenian-Croatian border, and during the following years, it emerged in more and more counties of northwestern Croatia. Surveillance of Ae. j. japonicus and other invasive mosquito species was subsequently extended both spatially and temporally in Croatia and neighbouring Bosnia and Herzegovina and Serbia. Mosquito collections were conducted in 2017 and 2018, based on adult trapping through dry ice-baited CDC traps and BG-Lure-baited BG-Sentinel traps, larval sampling through dippers and nets, and ovitrapping. Aedes j. japonicus specimens from collected samples were subjected to population genetic analysis by comparing microsatellite signatures and nad4 DNA sequences between sampled locations and with data previously obtained from more western European distribution areas. Aedes j. japonicus immature stages were found at 19 sites in Croatia, two sites in Bosnia and Herzegovina and one site in Serbia. In Croatia, four new counties were found colonised, two in the east and two in the south of the previously known distribution area. A spread of 250 km could thus be documented within five years. The findings in Bosnia and Herzegovina and Serbia represent the first records of Ae. j. japonicus in these countries. Genetic analysis suggests at least two introduction events into the surveyed area. Among the locations analysed, Orahovica can be considered a genetic border. The individuals collected west of this point were found to be similar to samples previously collected in the border regions of Southeast Germany/Austria and Austria/Slovenia, while the specimens from more eastern Croatian localities, together with those from Bosnia and Herzegovina and Serbia, were genetically different and could not be assigned to a probable origin. Thus, introduction from Central Europe, possibly by vehicular traffic, into the study area is likely, but other origins, transportation routes and modes of entry appear to contribute. Further dispersal of Ae. j. japonicus to other parts of southeastern Europe is anticipated.

An update of the Culicoides (Diptera: Ceratopogonidae) checklist for the Balkans.

BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: 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.

Distribution of Anopheles daciae and other Anopheles maculipennis complex species in Serbia.

Malaria is one of the most severe health problems facing the world today. Until the mid-twentieth century, Europe was an endemic area of malaria, with the Balkan countries being heavily infested. Sibling species belonging to the Anopheles maculipennis complex are well-known as effective vectors of Plasmodium in Europe. A vast number of human malaria cases in the past in the former Yugoslavia territory have stressed the significance of An. maculipennis complex species as primary and secondary vectors. Therefore, the present study evaluates the species composition, geographic distribution and abundance of these malaria vector species. Mosquitoes were collected in the northern Serbian province of Vojvodina and analysed by PCR-RFLP, multiplex PCR and sequencing of the ITS2 intron of genomic rDNA. Four sibling species of the An. maculipennis complex were identified. Both larvae and adults of the recently described species An. daciae were identified for the first time in Serbia. In 250 larval samples, 109 (44%) An. messeae, 90 (36%) An. maculipennis s.s., 33 (13%) An. daciae and 18 (7%) An. atroparvus were identified. In adult collections, 81 (47%) An. messeae, 55 (32%) An. daciae, 33 (19%) An. maculipennis s.s., and 3 (2%) An. atroparvus were recorded. The most abundant species in Vojvodina was An. messeae, whereas An. atroparvus was confirmed a rare species in all parts. Since this species is a potentially, highly competent malarial vector, low population density could be crucial to prevent a new establishment of endemic malaria transmission in Serbia.

West Nile virus 'circulation' in Vojvodina, Serbia: Mosquito, bird, horse and human surveillance.

Efforts to detect West Nile virus (WNV) in the Vojvodina province, northern Serbia, commenced with human and mosquito surveillance in 2005, followed by horse (2009) and wild bird (2012) surveillance. The knowledge obtained regarding WNV circulation, combined with the need for timely detection of virus activity and risk assessment resulted in the implementation of a national surveillance programme integrating mosquito, horse and bird surveillance in 2014. From 2013, the system showed highly satisfactory results in terms of area specificity (the capacity to indicate the spatial distribution of the risk for human cases of West Nile neuroinvasive disease - WNND) and sensitivity to detect virus circulation even at the enzootic level. A small number (n = 50) of Culex pipiens (pipiens and molestus biotypes, and their hybrids) females analysed per trap/night, combined with a high number of specimens in the sample, provided variable results in the early detection capacity at different administrative levels (NUTS2 versus NUTS3). The clustering of infected mosquitoes, horses, birds and human cases of WNND in 2014-2015 was highly significant, following the south-west to north-east direction in Vojvodina (NUTS2 administrative level). Human WNND cases grouped closest with infected mosquitoes in 2014, and with wild birds/mosquitoes in 2015. In 2014, sentinel horses showed better spatial correspondence with human WNND cases than sentinel chickens. Strong correlations were observed between the vector index values and the incidence of human WNND cases recorded at the NUTS2 and NUTS3 levels. From 2010, West Nile virus was detected in mosquitoes sampled at 43 different trap stations across Vojvodina. At 14 stations (32.56%), WNV was detected in two different (consecutive or alternate) years, at 2 stations in 3 different years, and in 1 station during 5 different years. Based on these results, integrated surveillance will be progressively improved to allow evidence-based adoption of preventive public health and mosquito control measures.

Choice of optimal biocide combination to control flies (Diptera: Muscidae).

INTRODUCTION: Flies - by feeding on decaying matter, human waste and food - have been implicated in the spread of numerous animal and human diseases. Excessive fly populations are generally associated with livestock units and domestic waste due to decaying organic matter. A large number of flies cause extreme disturbance in the behavior of the host, resulting in skin irritation, lesions, wounds, and secondary infections are likely to appear. OBJECTIVE: The aim of this study was to evaluate the effects of combined applications of larvicide (cyromazine) and adulticides (acetamiprid in formulation with pheromone and thiamethoxam) on the suppression of fly populations. MATERIALS AND METHODS: The study was conducted on a pig farm. The piglet farms are one of the most favorable places for fly breeding. Three units were used for biocide applications and a fourth unit as the control where biocides were not applied. The monitoring of pre- and post-treatment of adult fly populations was carried out by glued cardboards. The cards were hung on metal rods above piglet's cage. This monitoring method served as a parameter for the estimation of biological effectiveness. RESULTS: The highest degree of fly control (88.4% mortality 8 days after treatment) was achieved when a combination of cyromazine and thiamethoxam was used. A biocide based on sex pheromone (Z)-9-tricosene + acetamiprid was the most effective on flies 3 days after biocide application, with a mortality rate of 69.1 %. Thiamethoxam achieved the highest reduction of flies 6 days after treatment, with 78.19% obtained mortality. CONCLUSION: Biological efficacy of the applied biocides in combination ciromazine + thiamethoxam and thiamethoxam alone was justified.