Artificial resting sites: An alternative sampling method for adult mosquitoes.

Mosquito collections are commonly conducted with baited traps predominantly attracting host-seeking females. In contrast, resting sites are generally colonized by a broader range of the mosquito population, including a higher proportion of males and blood-engorged females. This study evaluates the sampling success of different artificial resting sites, attached to a deciduous or coniferous tree at different heights. As standard sampling method, carbon dioxide-baited Biogents Sentinel traps (BG traps) were operated in parallel. BG traps caught a higher number of specimens compared to the resting sites. However, the proportion of blood-engorged females and males was higher in resting sites. More Culiseta spp. specimens were collected in resting sites compared to BG traps, but less Aedes spp. specimens. In general, fewer specimens and species were recorded in small resting sites and at top height level compared to medium or large resting sites at medium or ground level. The proportion of males was highest at the ground, while the proportion of engorged females was highest at medium and top level. Due to the higher proportion of blood-engorged females, artificial resting sites are especially useful for studies of host-feeding patterns or xenosurveillance. Low costs and efforts allow a cost-effective increase of the number of resting sites per sampling site to collect more mosquitoes.

Physico-Chemical Characteristics of Culex pipiens sensu lato and Culex torrentium (Diptera: Culicidae) Breeding Sites in Germany.

Culex pipiens sensu lato (L.) and Culex torrentium Martini, 1925 are two widely distributed mosquito species in Europe. Due to difficulties in morphological discrimination, the current knowledge on differences in the breeding ecology is fragmentary. Therefore, this study evaluated the relation between the presence-absence of both species at various types of breeding habitats in response to physico-chemical parameters, using a recently developed molecular assay that allows reliable species-specific typing of larvae and pupae. The results revealed that the two species often occur in sympatry at the studied breeding sites, and there were no substantial differences concerning presence-absence of the two species with regard to the various environmental parameters investigated.

Topsoil conditions correlate with the emergence rates of Culicoides chiopterus and Culicoides dewulfi (Diptera: Ceratopogonidae) from cowpats.

Culicoides chiopterus (Meigen), 1830 and Culicoides dewulfi Goetghebuer, 1936 (Diptera: Ceratopogonidae) are considered to develop exclusively in dung, but do not necessarily show an equal distribution and abundance on livestock farms in Northern Europe. Recent modelling studies identified soil parameters to explain these differences. The present study addressed the question whether topsoil conditions underneath cowpats correlate with the number of emerging C. chiopterus and C. dewulfi. We recorded the emergence of biting midges from 24 cowpats over a period of 4 weeks and analysed samples from the topsoil. In agreement with species distribution models based on remote data, our results detected the correlation of soil moisture, organic matter and soil texture with the number of emerging C. chiopterus and C. dewulfi. With increasing soil moisture, the number of emerging adults increased for both species and the amount of organic matter was positively correlated with the number of emerging C. chiopterus. In contrast, soil textures showed conflicting results, i.e. a positive and negative relationship with the same variables. According to our results, soil underneath dung can explain the number of emerging Culicoides species. The knowledge of these effects might improve the interpretation of large-scaled distribution models for dung-breeding biting midges.

Culicoides biting midge density in relation to the position and substrate temperature in a cattle dung heap.

Dung heaps offer warm breeding sites, which might be suitable for a continuing development or even emergence at low air temperatures in winter. Therefore, this study collected substrate samples from the outer surface and core of a cattle dung heap at the beginning of the winter period. We aimed to analyze the density of immature Culicoides in relation to substrate position and temperature. We took samples from the outer layer and core of the dung heap at different heights. Floatation was used to extract Culicoides larvae from the dung heap samples. In order to rear larvae individually, we separated them in glass tubes. A total of 229 Culicoides larvae were extracted from the dung heap samples. Highest densities (99.1% of all larvae) were recorded for the outer layers of the dung heap but hardly any in the core (0.9% of all individuals). While the density of larvae was negatively correlated with increasing substrate temperatures, Culicoides larvae were found in a temperature range between 7.9 and 38.0 °C (mean 16.6 °C). Extracted larvae were reared to adults. All male individuals were identified as Culicoides obsoletus (Meigen), 1818 and all female individuals as C. obsoletus/Culicoides scoticus. It can be concluded that dung heaps offer temperature conditions, which allow the survival and probably also the development to adults for immature Culicoides also under harsh climate conditions in winter.

Impact of mechanical disturbance on the emergence of Culicoides from cowpats.

The outbreaks of Bluetongue virus and Schmallenberg virus between 2006 and 2012 highlighted the need for control methods for north-western Palaearctic biting midge species. There is especially a demand for alternative control methods without application of insecticides. Therefore, the objective of this paper was to evaluate the impact of mechanical disturbance as a control method for Culicoides biting midges in cowpats. We used a trash rake to destroy the compact structure of cowpats and spread the dung over the ground. We expected the disturbance to cause a faster alteration of physical characteristics and drying of the dung, resulting in lower emergence from disturbed cowpats. Emerging biting midges were collected with emergence traps. The number of emerged Culicoides was compared between treated and control cowpats. A total of 12,979 biting midges emerged, all belonging to the Obsoletus group. The majority of the 6,758 male individuals were identified as Culicoides chiopterus (Meigen, 1830) (91.4%), followed by Culicoides dewulfi (Goetghebuer, 1936) (6.5%), and a small number of Culicoides scoticus (Downes & Kettle, 1952) (1.2%). Our results showed no significant differences between the emergence of Culicoides (males and females) from disturbed and control cowpats. The lack of differences is discussed in light of the time period chosen for the experiment and the climatic conditions (especially low temperatures) during the study period. The climatic conditions during the study did not favour desiccation effects and therefore did not cause a severe alteration of the dung and the biotic parameters. In conclusion, immature Culicoides showed a high tolerance against mechanical disturbance.

A convolutional neural network to identify mosquito species (Diptera: Culicidae) of the genus Aedes by wing images.

Accurate species identification is crucial to assess the medical relevance of a mosquito specimen, but requires intensive experience of the observers and well-equipped laboratories. In this proof-of-concept study, we developed a convolutional neural network (CNN) to identify seven Aedes species by wing images, only. While previous studies used images of the whole mosquito body, the nearly two-dimensional wings may facilitate standardized image capture and reduce the complexity of the CNN implementation. Mosquitoes were sampled from different sites in Germany. Their wings were mounted and photographed with a professional stereomicroscope. The data set consisted of 1155 wing images from seven Aedes species as well as 554 wings from different non-Aedes mosquitoes. A CNN was trained to differentiate between Aedes and non-Aedes mosquitoes and to classify the seven Aedes species based on grayscale and RGB images. Image processing, data augmentation, training, validation and testing were conducted in python using deep-learning framework PyTorch. Our best-performing CNN configuration achieved a macro F1 score of 99% to discriminate Aedes from non-Aedes mosquito species. The mean macro F1 score to predict the Aedes species was 90% for grayscale images and 91% for RGB images. In conclusion, wing images are sufficient to identify mosquito species by CNNs.

Repeated introduction of Aedes albopictus into Germany, July to October 2012.

During a small-scale surveillance project to identify possible routes of entry for invasive mosquitoes into Germany, 14 adult Aedes (Stegomyia) albopictus (Skuse) were discovered between July and October 2012. They were trapped at three different service stations in Bavaria and Baden-Wuerttemberg located along two motorways that connect Germany with southern Europe. This indicates regular introduction of A. albopictus into Germany and highlights the need for a continuous surveillance and control programme.

Overwintering Culex torrentium in abandoned animal burrows as a reservoir for arboviruses in Central Europe.

Culex pipiens s.s./Culex torrentium belong to the most widespread mosquito taxa in Europe and are the main vectors of Sindbis, West Nile and Usutu virus. The adult overwintering females can act as reservoir for these arthropod-borne viruses (arboviruses), thus contributing to their local persistence when transmission cycles are interrupted during the winter. However, the main overwintering sites of Cx. torrentium are unknown. In a study from 2017, 3455 Cx. pipiens s.s./Cx. torrentium specimens were collected from abandoned animal burrows in Poznan, Poland. These specimens were retrospectively identified to species-level with a PCR assay, which revealed Cx. torrentium as dominant species (> 60%). Motivated by these results, we conducted a field study from February to July 2022 to systematically analyse the overwintering site patterns of Cx. pipiens s.s./Cx. torrentium. Mosquitoes were sampled using pipe traps in abandoned animal burrows (n = 20) and with aspirators in nearby anthropogenic overwintering sites (n = 23). All Cx. pipiens s.s./Cx. torrentium were screened for Flaviviridae RNA. In total, 4710 mosquitoes of five different taxa were collected from anthropogenic sites. 3977 of them were identified as Cx. p. pipiens/Cx. torrentium (Cx. p. pipiens: 85%, Cx torrentium: 1%, pools with both species: 14%). In contrast, only Cx. p. pipiens/Cx. torrentium (1688 specimens) were collected from animal burrows dominated by Cx. torrentium (52%), followed by pools with both species (40%) and Cx. p pipiens (8%). A single pool of 10 Cx. torrentium specimens collected from an animal burrow was positive for Usutu virus. The detection of Usutu virus demonstrates that Cx. torrentium can act as winter reservoir for arboviruses. Abandoned animal burrows may by the primary overwintering site for the species and should be considered in future surveillance programmes, when sampling overwintering mosquitoes.

Effects of mosquito resting site temperatures on the estimation of pathogen development rates in near-natural habitats in Germany.

BACKGROUND: Environmental temperature is a key driver for the transmission risk of mosquito-borne pathogens. Epidemiological models usually relate to temperature data from standardized weather stations, but these data may not capture the relevant scale where mosquitoes experience environmental temperatures. As mosquitoes are assumed to spend most of their lifetime in resting sites, we analysed mosquito resting site patterns and the associated temperatures in dependence on the resting site type, resting site height and the surrounding land use. METHODS: The study was conducted in 20 areas in near-natural habitats in Germany. Ten areas were studied in 2017, and another 10 in 2018. Each study area consisted of three sampling sites, where we collected mosquitoes and microclimatic data in artificial (= garden pop-up bags) and natural resting sites at three height levels between 0 and 6 m. Land use of the study sites was characterized as forest and meadows based on reclassified information of the CORINE (Coordination of Information on the Environment) Land Cover categories. The hourly resting site temperatures and the data from the nearest weather station of the German meteorological service were used to model the duration of the extrinsic incubation period (EIP) of mosquito-borne pathogens. RESULTS: Anopheles, Culex and Culiseta preferred artificial resting sites, while Aedes were predominantly collect in natural resting sites. Around 90% of the mosquitoes were collected from resting sites below 2 m. The mosquito species composition did not differ significantly between forest and meadow sites. Mean resting site temperatures near the ground were approximately 0.8 °C lower than at a height of 4-6 m, which changed the predicted mean EIP up to 5 days at meadow and 2 days at forest sites. Compared with temperature data from standardized weather stations, the resting site temperatures near the ground would prolong the mean estimated EIP 4 days at forest sites and 2 days at meadow sites. CONCLUSIONS: The microclimate of mosquito resting sites differs from standardized meteorological data, which can influence the transmission of mosquito-borne pathogens. In a near-natural environment, colder temperatures at mosquitoes' preferred resting sites near the ground would prolong the EIP of mosquito-borne pathogens relative to data from weather stations.

Modelling the monthly abundance of Culicoides biting midges in nine European countries using Random Forests machine learning.

BACKGROUND: Culicoides biting midges transmit viruses resulting in disease in ruminants and equids such as bluetongue, Schmallenberg disease and African horse sickness. In the past decades, these diseases have led to important economic losses for farmers in Europe. Vector abundance is a key factor in determining the risk of vector-borne disease spread and it is, therefore, important to predict the abundance of Culicoides species involved in the transmission of these pathogens. The objectives of this study were to model and map the monthly abundances of Culicoides in Europe. METHODS: We obtained entomological data from 904 farms in nine European countries (Spain, France, Germany, Switzerland, Austria, Poland, Denmark, Sweden and Norway) from 2007 to 2013. Using environmental and climatic predictors from satellite imagery and the machine learning technique Random Forests, we predicted the monthly average abundance at a 1 km2 resolution. We used independent test sets for validation and to assess model performance. RESULTS: The predictive power of the resulting models varied according to month and the Culicoides species/ensembles predicted. Model performance was lower for winter months. Performance was higher for the Obsoletus ensemble, followed by the Pulicaris ensemble, while the model for Culicoides imicola showed a poor performance. Distribution and abundance patterns corresponded well with the known distributions in Europe. The Random Forests model approach was able to distinguish differences in abundance between countries but was not able to predict vector abundance at individual farm level. CONCLUSIONS: The models and maps presented here represent an initial attempt to capture large scale geographical and temporal variations in Culicoides abundance. The models are a first step towards producing abundance inputs for R0 modelling of Culicoides-borne infections at a continental scale.

Monitoring of putative vectors of bluetongue virus serotype 8, Germany.

To identify the vectors of bluetongue virus (BTV) in Germany, we monitored Culicoides spp. biting midges during April 2007-May 2008. Molecular characterization of batches of midges that tested positive for BTV suggests C. obsoletus sensu stricto as a relevant vector of bluetongue disease in central Europe.

Monitoring of Culicoides at 20 locations in northwest Germany.

Entomological monitoring was carried out from April 2007 to May 2008 at 20 locations in the areas of Lower Saxony, Schleswig-Holstein, Hamburg, and Bremen. A total number of 26 Culicoides species were sampled by light traps during the first week of every month. Culicoides diversity was highest in summer, achieving more than 20 species and genera per month. Numbers of Culicoides were highest in spring and summer with a maximum of 325,000 individuals in May 2008 at a single location. During the winter, the number of individuals decreased considerably, but few individuals of Culicoides were present even during the coldest months in January and February with Culicoides obsoletus remaining the only species complex. The total number of Ceratopogonidae and the number of individuals from C. obsoletus complex and Culicoides pulicaris complex were significantly correlated with temperature almost at any date and location.

Bluetongue disease in Germany (2007-2008): monitoring of entomological aspects.

In the summer of 2006, a bluetongue epidemic started in the border area of Belgium, The Netherlands, and Germany, spread within 2 years over large areas of Western and Central Europe, and caused substantial losses in farm ruminants. Especially sheep and cattle were severely affected, leading to a case-fatality ratio of nearly 40% in sheep (Conraths et al., Emerg Inf Dis 15(3):433-435, 2009). The German federal ministry of food, agriculture, and consumer protection (BMELV) established a countrywide monitoring on the occurrence of the vectors of this virus, i.e., midges (family Ceratopogonidae) of the genus Culicoides. The monitoring was done on 91 sites, most of which were localized in the 150-km restriction zone that existed in December 2006. A grid consisting of 45 x 45 km(2) cells was formed that covered the monitoring area. As a rule, one trap was placed into each grid cell. The monitoring program started at the end of March 2007 and lasted until May 2008. It included the catching of midges by ultraviolet light traps-done each month from days 1 until 8, the selection of midges of the Culicoides obsoletus, Culicoides pulicaris group, and other Culicoides spp., the testing of midges for bluetongue virus (BTV) by polymerase chain reaction (PCR), and the daily registration of weather data at each trap site for the whole monitoring period. The following main results were obtained: (1) Members of the C. obsoletus group were most commonly found in the traps, reaching often 3/4 of the catches. The African and South European vector of BTV-the species Culicoides imicola-was never found. (2) Members of the C. obsoletus group were most frequently found infected with BTV besides a few cases in the C. pulicaris group and other species. (3) Members of the C. obsoletus group were also found in winter. Their numbers were reduced, however, and they were caught mostly close to stables. Therefore, a true midge-free period does not exist during the year in Germany. (4) The amounts of midges caught daily depended on the weather conditions. If it was cold and/or windy, the traps contained only a few specimens. Since the months from January to May 2008 were considerably colder (at all farms) than their correspondents in 2007, the growing of the population of midges started 2-3 months later in 2008 than in 2007. (5) The highest populations of midges occurred in both years (2007 and 2008) during the months September and October. This corresponded significantly to the finding of highest numbers of infected midges and to the number of diseased cattle and sheep during these 2 months. (6) It is noteworthy that in general, the first virus-positive midges of the species C. obsoletus were found about 1 1/2 months later than the first clinical cases had occurred or later than the first PCR-proven virus-positive sentinel animals had been documented. In 2007, the first BTV-positive cattle were detected in May in North Rhine-Westphalia, while the first positive Culicoides specimens were only found in August on the same farm. Evaluating these main results of the entomological monitoring and the fact that many wild ruminants have also been infected with BTV, it becomes evident that bluetongue disease has become endemic in Central Europe, and that only constant effort including vaccination and perhaps also insecticidal protection of cattle and sheep will keep the economical losses at a reasonable level. The following papers (1-10) in this journal will contribute more details obtained from this worldwide unique entomological monitoring: Bartsch et al. 2009; Bauer et al. 2009; Stephan et al. 2009; Clausen et al. 2009; Hörbrand and Geier 2009; Kiehl et al. 2009; Mehlhorn et al. 2009; Kiel et al. 2009; Vorsprach et al. 2009; Balczun et al. 2009.

Comparative analysis of subsampling methods for large mosquito samples.

BACKGROUND: The analysis of large mosquito samples is expensive and time-consuming, delaying the efficient timing of vector control measurements. Processing a fraction of a sample using a subsampling method can significantly reduce the processing effort. However, a comprehensive evaluation of the reliability of different subsampling methods is missing. METHODS: A total of 23 large mosquito samples (397-4713 specimens per sample) were compared in order to evaluate five subsampling methods for the estimation of the number of specimens and species: area, volume, weight, selection of 200 random specimens and analyses with an image processing software. Each sample was distributed over a grid paper (21.0 × 29.7 cm; 25 grid cells of 4.2 × 5.9 cm) with 200 randomly distributed points. After taking pictures, mosquito specimens closest to each of the 200 points on the paper were selected. All mosquitoes per grid cell were identified by morphology and transferred to scaled tubes to estimate the volume. Finally, the fresh and dry weights were determined. RESULTS: The estimated number of specimens and species did not differ between the area-, volume- and weight-based method. Subsampling 20% of the sample gave an error rate of approximately 12% for the number of specimens, 6% for the proportion of the most abundant species and between 6-40% for the number of species per sample. The error for the estimated number of specimens using the picture processing software ImageJ gave a similar error rate when analyzing 15-20% of the total sample. By using 200 randomly selected specimens it was possible to give a precise estimation of the proportion of the most abundant species (r = 0.97, P < 0.001), but the number of species per sample was underestimated by 28% on average. Selecting adjacent grid cells instead of sampling randomly chosen grid cells and using dry weight instead of wet weight did not increase the accuracy of estimates. CONCLUSIONS: Different subsampling methods have various advantages and disadvantages. However, the area-based analysis of 20% of the sample is probably the most suitable approach for most kinds of mosquito studies, giving sufficiently precise estimations of the number of specimens and species, which is slightly less laborious compared to the other methods tested.

Reintroduction of freshwater macroinvertebrates: challenges and opportunities.

Species reintroductions - the translocation of individuals to areas in which a species has been extirpated with the aim of re-establishing a self-sustaining population - have become a widespread practice in conservation biology. Reintroduction projects have tended to focus on terrestrial vertebrates and, to a lesser extent, fishes. Much less effort has been devoted to the reintroduction of invertebrates into restored freshwater habitats. Yet, reintroductions may improve restoration outcomes in regions where impoverished regional species pools limit the self-recolonisation of restored freshwaters. We review the available literature on macroinvertebrate reintroductions, focusing on identifying the intrinsic and extrinsic factors that determine their success or failure. Our study reveals that freshwater macroinvertebrate reintroductions remain rare, are often published in the grey literature and, of the attempts made, approximately one-third fail. We identify life-cycle complexity and remaining stressors as the two factors most likely to affect reintroduction success, illustrating the unique challenges of freshwater macroinvertebrate reintroductions. Consideration of these factors by managers during the planning process and proper documentation - even if a project fails - may increase the likelihood of successful outcomes in future reintroduction attempts of freshwater macroinvertebrates.

Spatial and temporal variation in the abundance of Culicoides biting midges (Diptera: Ceratopogonidae) in nine European countries.

BACKGROUND: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are vectors of bluetongue virus (BTV), African horse sickness virus and Schmallenberg virus (SBV). Outbreaks of both BTV and SBV have affected large parts of Europe. The spread of these diseases depends largely on vector distribution and abundance. The aim of this analysis was to identify and quantify major spatial patterns and temporal trends in the distribution and seasonal variation of observed Culicoides abundance in nine countries in Europe. METHODS: We gathered existing Culicoides data from Spain, France, Germany, Switzerland, Austria, Denmark, Sweden, Norway and Poland. In total, 31,429 Culicoides trap collections were available from 904 ruminant farms across these countries between 2007 and 2013. RESULTS: The Obsoletus ensemble was distributed widely in Europe and accounted for 83% of all 8,842,998 Culicoides specimens in the dataset, with the highest mean monthly abundance recorded in France, Germany and southern Norway. The Pulicaris ensemble accounted for only 12% of the specimens and had a relatively southerly and easterly spatial distribution compared to the Obsoletus ensemble. Culicoides imicola Kieffer was only found in Spain and the southernmost part of France. There was a clear spatial trend in the accumulated annual abundance from southern to northern Europe, with the Obsoletus ensemble steadily increasing from 4000 per year in southern Europe to 500,000 in Scandinavia. The Pulicaris ensemble showed a very different pattern, with an increase in the accumulated annual abundance from 1600 in Spain, peaking at 41,000 in northern Germany and then decreasing again toward northern latitudes. For the two species ensembles and C. imicola, the season began between January and April, with later start dates and increasingly shorter vector seasons at more northerly latitudes. CONCLUSION: We present the first maps of seasonal Culicoides abundance in large parts of Europe covering a gradient from southern Spain to northern Scandinavia. The identified temporal trends and spatial patterns are useful for planning the allocation of resources for international prevention and surveillance programmes in the European Union.

Monthly variation in the probability of presence of adult Culicoides populations in nine European countries and the implications for targeted surveillance.

BACKGROUND: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are small hematophagous insects responsible for the transmission of bluetongue virus, Schmallenberg virus and African horse sickness virus to wild and domestic ruminants and equids. Outbreaks of these viruses have caused economic damage within the European Union. The spatio-temporal distribution of biting midges is a key factor in identifying areas with the potential for disease spread. The aim of this study was to identify and map areas of neglectable adult activity for each month in an average year. Average monthly risk maps can be used as a tool when allocating resources for surveillance and control programs within Europe. METHODS: We modelled the occurrence of C. imicola and the Obsoletus and Pulicaris ensembles using existing entomological surveillance data from Spain, France, Germany, Switzerland, Austria, Denmark, Sweden, Norway and Poland. The monthly probability of each vector species and ensembles being present in Europe based on climatic and environmental input variables was estimated with the machine learning technique Random Forest. Subsequently, the monthly probability was classified into three classes: Absence, Presence and Uncertain status. These three classes are useful for mapping areas of no risk, areas of high-risk targeted for animal movement restrictions, and areas with an uncertain status that need active entomological surveillance to determine whether or not vectors are present. RESULTS: The distribution of Culicoides species ensembles were in agreement with their previously reported distribution in Europe. The Random Forest models were very accurate in predicting the probability of presence for C. imicola (mean AUC = 0.95), less accurate for the Obsoletus ensemble (mean AUC = 0.84), while the lowest accuracy was found for the Pulicaris ensemble (mean AUC = 0.71). The most important environmental variables in the models were related to temperature and precipitation for all three groups. CONCLUSIONS: The duration periods with low or null adult activity can be derived from the associated monthly distribution maps, and it was also possible to identify and map areas with uncertain predictions. In the absence of ongoing vector surveillance, these maps can be used by veterinary authorities to classify areas as likely vector-free or as likely risk areas from southern Spain to northern Sweden with acceptable precision. The maps can also focus costly entomological surveillance to seasons and areas where the predictions and vector-free status remain uncertain.

Comparison of single- and multi-scale models for the prediction of the Culicoides biting midge distribution in Germany.

This study analysed Culicoides presence-absence data from 46 sampling sites in Germany, where monitoring was carried out from April 2007 until May 2008. Culicoides presence-absence data were analysed in relation to land cover data, in order to study whether the prevalence of biting midges is correlated to land cover data with respect to the trapping sites. We differentiated eight scales, i.e. buffer zones with radii of 0.5, 1, 2, 3, 4, 5, 7.5 and 10 km, around each site, and chose several land cover variables. For each species, we built eight single-scale models (i.e. predictor variables from one of the eight scales for each model) based on averaged, generalised linear models and two multiscale models (i.e. predictor variables from all of the eight scales) based on averaged, generalised linear models and generalised linear models with random forest variable selection. There were no significant differences between performance indicators of models built with land cover data from different buffer zones around the trapping sites. However, the overall performance of multi-scale models was higher than the alternatives. Furthermore, these models mostly achieved the best performance for the different species using the index area under the receiver operating characteristic curve. However, as also presented in this study, the relevance of the different variables could significantly differ between various scales, including the number of species affected and the positive or negative direction. This is an even more severe problem if multi-scale models are concerned, in which one model can have the same variable at different scales but with different directions, i.e. negative and positive direction of the same variable at different scales. However, multi-scale modelling is a promising approach to model the distribution of Culicoides species, accounting much more for the ecology of biting midges, which uses different resources (breeding sites, hosts, etc.) at different scales.

Effects of temperature and photoperiod on the development of overwintering immature Culicoides chiopterus and C. dewulfi.

In areas with harsh winters, several studies have recorded low numbers - or even zero - adult Culicoides during the winter period followed by a population peak in the subsequent spring. This experimental study determined whether temperature, photoperiod, or the combination thereof affect the development of overwintering immature dung breeding Culicoides, resulting in this peak. Temperature had a significant impact on the development period of Culicoides chiopterus (Meigen), 1830 and C. dewulfi Goetghebuer, 1936. In treatments with constant high temperature, emergence occurred shortly after the beginning of the experiment (mean=day 9). In contrast, no individuals emerged from the other two treatments, as long as the temperatures were below 10°C. In these treatments, the emergence of Culicoides started when the temperature exceeded 20°C for some days (mean=day 33). There was no significant difference between the two photoperiods (February or April day length; 9 h:15 h [light:dark] vs. 13 h:11 h). Our results highlight the importance of temperature on the spring emergence of C. chiopterus and C. dewulfi, but the response to the four temperature-photoperiod treatments did not differ between the two species.