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To investigate the role of dipterans in the transmission of Onchocerca lupi and other zoonotic filarioids, samples were collected from different sites in Algarve, southern Portugal, morphologically identified and molecularly tested for filarioids. Culex sp. (72.8%) represented the predominant genus followed by Culicoides sp. (11.8%), Ochlerotatus sp. (9.7%), Culiseta sp. (4.5%), Aedes sp. (0.9%) and Anopheles sp. (0.3%). Nineteen (2.8%) specimens scored positive for filarioids, with Culex pipiens quinquefasciatus (2%) positive for Dirofilaria immitis (1.4%), Dirofilaria repens, Acanthocheilonema reconditum, Onchocerca lupi, unidentified species of Filarioidea (0.2%, each) and Onchocercidae (0.6%). Additionally, Culiseta longiareolata (6.5%), Ochlerotatus caspius (3%) and Culex laticinctus (0.2%) scored positive for unidentified Onchocercidae, A. reconditum and for O. lupi, respectively. This is the first report of the occurrence of DNA of O. lupi, D. repens and A. reconditum in Culex spp. in Portugal. Information regarding the vectors and the pathogens they transmit may help to adopt proper prophylactic and control measures., First report of the occurrence of the DNA of Onchocerca lupi, Dirofilaria repens and Acanthocheilonema reconditum in Culex spp. from Portugal.Unidentified species of Filarioidea and Onchocercidae were detected in mosquitoes from Portugal.The detection of Wolbachia supergroup E in Culex laticinctus positive for O. lupi is new to science and deserves further investigation.

Culicoides Latreille (Diptera: Ceratopogonidae) transmit arboviruses affecting wild and domestic ruminants such as bluetongue (BTV) and Schmallenberg virus (SBV). The sub-adult development and lifespan of Culicoides obsoletus s.s. (Meigen), Culicoides circumscriptus Kieffer and Culicoides paolae Boorman were examined at three different temperatures under laboratory conditions. Insects were collected from field between spring and autumn 2015 in two livestock farms located in Majorca (Spain). Gravid females were held individually at 18, 25 or 30 °C. Low temperatures increased the adult lifespan, time to oviposit and rate of development, whereas high temperatures increased the number of eggs, successful pupation and adult emergence as well as the larvae growth rate. The results showed that C. obsoletus s.s. have optimum development at 18 °C, whereas the optimal rearing temperature for C. circumscriptus and C. paolae was under warmer conditions of 25-30 °C. Variations in temperature/humidity and assays with different materials and substrates for oviposition should be considered in future studies. Understanding the requirements of the different species of Culicoides optimizing the results should be of special interest for predicting environmental change effects on these species, in addition to determining the rearing conditions for candidate Culicoides vectors.

Background Culicoides obsoletus is an important pest for animal husbandry. In the world, it is a major vector of bluetongue disease, which has been added to the list of Class A Animal Diseases by the World Organization for Animal Health (OIE) and must be reported in the event of an outbreak. Although numerous studies on Culicoides sampling in farms exist, the distribution of C. obsoletus in different landscapes remains unclear. Results This study aimed to assess the effect of landscape type on the distribution of C. obsoletus. Samples were collected using light traps during spring and fall. The effect of landscape type on the distribution of C. obsoletus samples was measured by the maximum entropy model (MaxEnt). Results show that C. obsoletus mainly appears in forests and pastures. Conclusion Our results may provide references for controlling the C. obsoletus population and provide suggestions of other areas that may also face the challenge of bluetongue.

The diversity and abundance of Culicoides (Diptera: Ceratopogonidae) and mosquitoes (Diptera: Culicidae) were studied in three animal protection centres (APCs) in Northern Spain between 1 July and 31 October 2018. Four miniature suction CDC light traps (two UV and two standard incandescent bulb traps, both types baited and non-baited with CO2 ) were placed in each APC to compare their efficiency in the collection of these Diptera groups. A total of 1176 biting midges (14 species), 224 mosquitoes (8 species) and 1 black fly were collected and identified by both morphological and molecular approaches. The Culicoides obsoletus complex (C. obsoletus/C. scoticus) accounted for 58.2% of the total collection within the Ceratopogonidae family, whereas Culex pipiens/Cx. torrentium comprised 76.8% of the Culicidae. The input of CO2 in light traps proved largely ineffective in improving the collections of both Diptera groups. UV-light traps were 7.8 and 2.2 times more effective than incandescent light traps in trapping Culicoides and mosquitoes, respectively. Seasonal dynamics differed between both Diptera taxa but captures of both taxa were significantly larger at the beginning of the summer. The epidemiological relevance of the most prevalent species is also discussed.

The effect of climatic factors on the presence of Culicoides Latreille (Diptera: Ceratopogonidae) was previously studied. Nevertheless, lack of laboratory rearing data hampers species-specific prediction of weather fluctuations effect on population size. To determine fluctuations in population size in the field, we recorded Culicoides and other Nematocerans in seven Israeli dairy farms over two-years (2011-2012) and analysed the association of their dynamics with fluctuations in ambient temperature and total rainfall. In six farms, the most abundant species were Culicoides imicola Kieffer and Culicoides schultzei (Enderlein) gp., primarily composed of parous females, and in one farm Culicoides obsoletus (Meigen) gp., mostly nulliparous females, were dominant. While the total number of insects was similar in both years, Culicoides numbers were significantly higher in 2012, but appeared later in the season and reached a higher peak. A multi-variable linear regression model demonstrated positive association of C. imicola and C. schultzei numbers with the monthly multi-annual ambient temperature and its specific deviation, but not with monthly rainfall. C. obsoletus populations peaked at spring and sharply decreased when temperature exceeded 20 °C, and were best modelled by adding quadratic terms. Weather-specific estimation of population size under field conditions may enable to predict outbreaks intensity of Culicoides-borne viruses.

Climatic, landscape, and host features are critical components in shaping outbreaks of vector-borne diseases. However, the relationship between the outbreaks of vector-borne pathogens and their environmental drivers is typically complicated, nonlinear, and may vary by taxonomic units below the species level (e.g., strain or serotype). Here, we aim to untangle how these complex forces shape the risk of outbreaks of Bluetongue virus (BTV); a vector-borne pathogen that is continuously emerging and re-emerging across Europe, with severe economic implications. We tested if the ecological predictors of BTV outbreak risk were serotype-specific by examining the most prevalent serotypes recorded in Europe (1, 4, and 8). We used a robust machine learning (ML) pipeline and 23 relevant environmental features to fit predictive models to 24,245 outbreaks reported in 25 European countries between 2000 and 2019. Our ML models demonstrated high predictive performance for all BTV serotypes (accuracies > 0.87) and revealed strong nonlinear relationships between BTV outbreak risk and environmental and host features. Serotype-specific analysis suggests, however, that each of the major serotypes (1, 4, and 8) had a unique outbreak risk profile. For example, temperature and midge abundance were as the most important characteristics shaping serotype 1, whereas for serotype 4 goat density and temperature were more important. We were also able to identify strong interactive effects between environmental and host characteristics that were also serotype specific. Our ML pipeline was able to reveal more in-depth insights into the complex epidemiology of BTVs and can guide policymakers in intervention strategies to help reduce the economic implications and social cost of this important pathogen.

Equine insect bite hypersensitivity (IBH) is a pruritic skin allergy caused primarily by biting midges, Culicoides spp. IBH susceptibility has polygenic inheritance and occurs at high frequencies in several horse breeds worldwide, causing increased costs and reduced welfare of affected horses. The aim of this study was to identify and validate single nucleotide polymorphisms (SNPs) associated with equine IBH susceptibility. After quality control, 33,523 SNPs were included in a Bayesian genome-wide association study on 177 affected and 178 unaffected Icelandic horses. We report associated regions in E. caballus (ECA) 1, 3, 15 and 18, overlapping with known IBH QTLs in horses, and novel regions containing several genes, together explaining 11.46% of the total genetic variance. For validation, three SNPs on ECA 1 and ECA X (explaining the largest percentage of genetic variance) within 1-mb genomic windows for IBH were genotyped in an independent population of 280 Exmoor ponies. The associated genomic region (152-153 mb) on ECA 1 was confirmed in Exmoor ponies and contains the AQR gene involved in splicing processes and a long non-coding RNA. This study confirms the polygenic nature of IBH susceptibility and suggests a role of transcriptional regulatory mechanisms (e.g., alternative splicing) for IBH predisposition in these horse breeds. ispartof: JOURNAL OF ANIMAL BREEDING AND GENETICS vol:137 issue:2 pages:223-233 ispartof: location:Germany status: published

In a controlled field trial, a mechanical vacuum and a mouth aspirator were compared with respect to Culicoides midge catching results. The two collection methods were equally applied to two Haflinger horses. Once every hour between 2 hr before sunset and sunset, midges were aspirated for 3 min directly off from the coat of each horse at the same time. In total, 16 replicate measurement days were made. To get insight into the number of (blood‐fed) midges collected on different body parts, four replicate measurement days were done in which the hourly 3‐min samples for each collecting method were separately collected and counted per body part. Mean number of Culicoides midges collected on the legs and on the belly of the horse was two times and significantly higher by the mechanical vacuum aspirator compared to the mouth aspirator. Although the mean number of Culicoides midges from the other part of the host sampled (head/neck/manes/back/flanks) was comparable between the two methods, the mean number of blood‐fed Culicoides midges was four times and significantly higher collected by the mouth aspirator compared to the mechanical vacuum aspirator. In conclusion, it can be recommended to use a mechanical vacuum aspirator instead of a mouth aspirator for the legs and belly area, where a systematic, stroke‐by‐stroke‐vacuum procedure is recommended. For the rest of the body, it is strongly recommended to use a targeted procedure by visual guidance of the presence of midges. For now, based on this experiment, we can only recommend using the mouth aspirator for this targeted approach. However, it might be worth assessing the use of the mechanical vacuum aspirator in a targeted way for future studies.

An upsurge in African horse sickness (AHS) in the Eastern Cape, South Africa, from 2006 led to an epidemiological reassessment of the disease there. Light trapping surveys carried out near horses, donkeys and zebras in 2014-2016 collected 39 species of Culicoides midge (Diptera: Ceratopogonidae) that are potential vectors of AHS. To establish if these midges fed on equids, DNA sequences were obtained from the gut contents of 52 female midges (35 freshly blood-fed, 13 gravid and four parous), representing 11 species collected across 11 sites. Culicoides leucostictus fed on all three equids. Culicoides bolitinos, Culicoides imicola and Culicoides magnus fed on both horses and donkeys. Culicoides onderstepoortensis fed on donkeys, and Culicoides similis and Culicoides pycnostictus fed on zebras. Bloodmeals from cows, pigs, warthogs, impalas and a domestic dog were also identified in various species, but none of the midges tested had fed on birds. These results contribute to knowledge of the vectorial capacity of several species of Culicoides with regard to AHS in the Eastern Cape and point to potential reservoir hosts, of which donkeys, zebras and domestic dogs have previously been found to harbour AHS. Blood-fed midges were also obtained throughout winter, indicating the potential for endemic AHS in the province.

Simbu serogroup are arbo‐ viruses which are mainly transmitted by Culicoides. Two members of the Simbu serogroup, Akabane and Shuni viruses, have been isolated from congenitally malformed ruminants in Israel. A recent serosurvey revealed that Israeli ruminants have been exposed to several additional Simbu viruses, including Shamonda and Sathuperi that seems to be circulating in Israel. In April 2017, an apparently healthy one‐month‐old male calf was transferred to the Kimron Veterinary Institute. A few days later, the calf was reported to be slow to respond to its surroundings and was not able to feed on its own. Blindness was observed upon clinical examination. RNA of the small, medium and large segments of Simbu serogroup viruses were amplified and sequenced from the testis tissues and from the Cerebrospinal fluid (CSF). During post‐mortem examination, hydranencephaly was defined. Phylogenetic analysis of all three segments of Simbu serogroup viruses showed that the sequences detected in the Israeli calf were virtually identical to Peaton virus (PEAV). PEAV was also detected in two pools of Culicoides imicola trapped at two different locations in Israel. This is the first genomic detection of PEAV outside Australia and Japan. These results are of epidemiological significance, as they demonstrate that PEAV is circulating in Israel and affects cattle. Consequently, these results are also of relevance to a potential spread of Simbu serogroup viruses into Europe.

BACKGROUND Genetic, epidemiologic, and clinical evidence suggests that, in horses, there are manifestations of hypersensitivity that can occur together. OBJECTIVES To investigate whether concurrent insect bite hypersensitivity (IBH) and severe equine asthma (EA) is associated with higher allergen-specific and total serum immunoglobulin E (IgE) concentrations than only EA or IBH. ANIMALS Healthy control horses (C, n = 40), horses with IBH (IBH, n = 24), severe EA (EA, n = 18), and both conditions (IBH/EA, n = 23) were included. METHODS In our retrospective comparative study, sera from horses with signs of severe EA, IBH, and control animals were used. IgE specific for 15 recombinant (r) allergens as well as total serum IgE concentrations were measured by enzyme-linked immunosorbent assay. RESULTS Group IBH (median sum r-Culicoides IgE: optical density at 405 nm [OD ] = 3.54 [0.48-15.07]) and group IBH/EA (OD = 4.55 [0.46-17.15]) had significantly (P .05) differences between group IBH and group IBH/EA. No significant differences among the groups were found for the other r-allergens or total serum IgE concentration. Compared to controls, horses with severe IBH had significantly increased IgE concentration to 5 Culicoides r-allergens (P

While wing form is known to differ between males and females of the genus Culicoides, detailed studies of sexual dimorphism are lacking. In this study, we analyze sex-specific differences in the wing form of 5 species of the subgenus Avaritia, using geometric morphometrics and comparative phylogenetic methods. Our results confirm the existence of marked sexual dimorphism in the wing form of the studied species and reveal for the first time that while there is a shared general pattern of sexual shape dimorphism within the subgenus, sexual size dimorphism, and particular features of sexual shape dimorphism differ among species. Sexual shape dimorphism was found to be poorly associated to size and the evolutionary history of the species. The tight association of sexual shape dimorphism with aspect ratio suggests that the shape of the wing is optimized for the type of flight of each sex, that is, dispersal flight in females versus aerobatic flight in males. Moreover, the fact that interspecific shape differences are greater and more strongly associated to aspect ratio in males than in females might be indicating that in males the selective pressures affecting flight performance characteristics are more heterogeneous and/or stronger than in females among the studied species. info:eu-repo/semantics/acceptedVersion

We investigated the floral characteristics, floral biology and floral visitors of the six Bornean Tacca species: T. bibracteata (only floral characteristics), T. borneensis, T. havilandii, T. leontopetaloides, T. palmata and T. reducta, and T. cristata from Peninsular Malaysia. All species are protogynous with pollen strings extruded post flower opening. Blooming of all species started from dawn except for T. leontopetaloides which flowered from dusk. While T. borneensis, T. cristata, T. havilandii, T. leontopetaloides and T. reducta are facultatively autogamic as the pollen/ovule ratios (P/O ratios) were low, T. bibracteata is facultatively xenogamic as its P/O ratio was higher. Four species (T. borneensis, T. cristata, T. havilandii and T. reducta) were tested for autonomous self‐pollination but all failed to set fruit. Manual self‐ and cross‐pollination treatments of T. borneensis, T. cristata and T. havilandii showed reduced fruit set and seed set. In contrast, T. reducta was highly self‐compatible. The showy bracts and bracteoles of T. borneensis are needed to guarantee pollination success but is not so in the other species investigated. Tacca are pollinated by two pollination guilds of female midges: two species of Forcipomyia (Lasiohelea) and Culicoides hinnoi. The floral biology and mating system of Tacca species indicate that most fruits and seeds were produced in samples resulting from natural pollination.

Biting midges of the genus Culicoides are known vectors of arboviruses affecting human and animal health. However, little is known about Culicoides imicola microbiota and its influence on this insect's biology. In this study, the impact of biotic and abiotic factors on C. imicola microbiota was characterized using shotgun-metagenomic sequencing of whole-body DNA samples. Wild-caught C. imicola adult nulliparous females were sampled in two locations from Sicily, Italy. The climatic variables of temperature and soil moisture from both localities were recorded together with potential host bloodmeal sources. Shared core microbiome among C. imicola populations included Pseudomonas, Escherichia, Halomonas, Candidatus Zinderia, Propionibacterium, and Schizosaccharomyces. Specific and unique taxa were also found in C. imicola from each location, highlighting similarities and differences in microbiome composition between the two populations. DNA and protein identification showed differences in host preferences between the two populations, with Homo sapiens and Canis lupus familiaris L. being the preferred bloodmeal source in both locations. A principal component analysis showed that the combined effect of host preferences (H. sapiens) and local soil moisture factors shape the microbiome composition of wild-caught populations of C. imicola. These results contribute to characterizing the role of the microbiome in insect adaptation and its utility in predicting geographic expansion of Culicoides species with potential implications for the control of vector-borne diseases.

Culicoides biting midges (Diptera: Ceratopogonidae) are vectors of a range of orbiviruses that cause important veterinary diseases such as bluetongue and African horse sickness. The effective monitoring of Culicoides species diversity and abundance, both at livestock and near potential wildlife hosts, is essential for risk management. The Onderstepoort 220-V ultraviolet (UV) light trap is extensively used for this purpose. Reducing its power requirements by fitting low-energy light-emitting diodes (LEDs) can lead to greater flexibility in monitoring. A comparison of the efficiency of the 220-V Onderstepoort trap (8-W fluorescent UV light) with the efficiency of the 220-V or 12-V Onderstepoort traps fitted with red, white, blue or green LEDs or a 12-V fluorescent Onderstepoort trap demonstrated the 220-V Onderstepoort trap to be the most efficient. All the results showed nulliparous Culicoides imicola Kieffer females to be the dominant grouping. Despite the lower numbers collected, 12-V traps can be used in field situations to determine the most abundant species.

To determine relative host preference rates and to establish whether a pair of animals in close proximity (between–host interaction) modified Culicoides attack and abundance profiles, compared to those tethered in isolation (host independence), Culicoides midges were pooted hourly from two sets of experimental animals: (1) a heifer cow and Fjord horse tethered close together (5 m apart), and (2) a heifer and Fjord tethered in isolation (45 m apart). Over 12 days, 570 3-min observations yielded 23 090 midges, representing 24 species. Approximately 95% belonged to the Culicoides obsoletus (Meigen) complex (two species), Culicoides dewulfi Goetghebuer, Culicoides chiopterus (Meigen), Culicoides punctatus (Meigen), Culicoides pulicaris (L.), and Culicoides achrayi Kettle & Lawson (Diptera: Ceratopogonidae) combined. There was no evidence for between-host interaction. Mean Culicoides species-specific attack rates did not differ between animal species, except that C. chiopterus was 7× more abundant on the legs of the heifer compared to the horse, and C. dewulfi twice as abundant on the upper half of the horse compared to the heifer. By contrast, mean species–specific biting rates of the C. obsoletus complex, C. chiopterus, C. dewulfi, C. punctatus, and C. pulicaris midges were 5×, 100×, 1.7×, 2×, and 2.5× lower in the horses compared to the heifers, respectively. It is not clear why high Culicoides attack rates observed in the horses do not convert into high biting rates as seen in cattle; this should be a subject for future research. In light of its apparent predilection for equids, the ability of C. dewulfi to replicate African horse sickness virus (AHSV) extrinsically should be investigated in the laboratory.

This research contributes to knowledge of the basic bionomic parameters of vector and non-vector Culicoides species. Field-collected gravid C. imicola and Obsoletus complex showed the longest lifespans in laboratory conditions. Culicoides paolae and C. circumscriptus seemed to be the most suitable species for laboratory rearing in view of their high oviposition rates, short lifecycles, long adult lifespans and female-biased sex ratios.

Akabane virus (AKAV) is teratogenic to the foetus of domestic ruminants and causes a significant reproduction loss in cattle in Japan. In several past epizootics in cattle, AKAV was also associated with post-natal encephalomyelitis, mainly in calves and young stock. Previously analysed AKAV isolates in East Asia form two major clusters, genogroups I and II, with isolates involved in encephalomyelitis belonging mainly to the former. Between 2007 and 2013, AKAV epizootics were regularly observed in Japan during the summer/autumn season, and abnormal deliveries and post-natal encephalomyelitis caused by the virus in cattle were reported. During this period, 30 AKAV isolates were obtained from diseased and sentinel cattle, a piglet and Culicoides biting midges throughout Japan and were subjected to genetic comparison and phylogenetic analysis with previous isolates. In 2007, 2011 and 2013, AKAV belonging to genogroup I was identified in the central nervous systems of calves showing neurological disorders. Notably, a total of 165 cases of bovine encephalomyelitis were reported in 2011 and the isolated viruses from affected animals shared high genetic identities with a South Korean isolate that was associated with a large outbreak in 2010, suggesting some epidemiological linkage between these epizootics. Epizootics of genogroup II were observed in 2008 and 2010, but bovine post-natal encephalomyelitis cases rarely occurred. Our findings suggest that the frequent incursion of genogroup I isolates has increased the frequency of post-natal encephalomyelitis cases in Japan in recent years. Infection by genogroup I virus was also identified in piglets with neurological disorders or congenital malformations in 2011 and 2013. The aetiological role of AKAV in pigs should be elucidated in the future.

Several species of Culicoides (Diptera: Ceratopogonidae) are vectors of pathogens, such as the bluetongue (BTV) and Schmallenberg (SBV) viruses, which cause important diseases in domestic and wild ruminants. As wild ruminants can contribute to overwintering and epizootics of both diseases, knowledge of the host-feeding behaviour of Culicoides in natural ecosystems is important to better understand their epidemiology. Blood-engorged Culicoides females trapped in natural areas inhabited by different wild ruminant species were genetically analysed to identify host species. The origin of bloodmeals was identified in 114 females of 14 species of Culicoides. A total of 104 (91.1%) Culicoides fed on mammals and 10 (8.9%) on birds. The most abundant host identified was red deer (66.7%), followed by humans (13%) and fallow deer (6.1%). Eleven of the 14 species of Culicoides fed exclusively on mammalian hosts. Among them, five are mammalophilic species considered to be important BTV and/or SBV vectors. The results of the present study confirm that Culicoides imicola, Culicoides obsoletus, Culicoides scoticus, Culicoides pulicaris and Culicoides punctatus fed on wild ruminants, and therefore support the hypothesis that these species can act as bridge vectors by facilitating the circulation of pathogens between wild and domestic ruminant communities.

Biting midges, Culicoides spp. (Diptera: Ceratopogonidae), are important vectors of viral pathogens. Following the outbreak of bluetongue serotype 8 in Europe between 2006 and 2009, many Culicoides surveillance programmes were initiated to identify vector-active periods, in accordance with European Commission regulation 2007/1266/EC. This study utilized surveillance data from 4 years of continuous light-trapping at 14 sites in Northern Ireland. The number of captured Culicoides varied from none during the vector-free period (December–April) to more than 36 000 per night during peak activity in the summer. The Obsoletus group represented 75% of Culicoides collected and the Pulicaris group represented 21%. A total of 91% of Culicoides were female, of which 42% were parous. Abundance data, sex ratios and parous rates suggested that both the Obsoletus and Pulicaris groups underwent three generations/year. The Obsoletus group was associated with cattle-rearing habitats and woodland, the Impunctatus group was found in habitats related to sheep rearing and the Pulicaris group were associated with both cattle and sheep. Housing did not reduce incursion of female Obsoletus group Culicoides but it did for males and for the Pulicaris group Culicoides. The influence of housing was strongly affected by time of year, probably reflecting the presence of livestock indoors/outdoors.

Culicoides (Diptera: Ceratopogonidae) are vectors of pathogens that affect wildlife, livestock and, occasionally, humans. Culicoides imicola (Kieffer, 1913) is considered to be the main vector of the pathogens that cause bluetongue disease (BT) and African horse sickness (AHS) in southern Europe. The study of blood-feeding patterns in Culicoides is an essential step towards understanding the epidemiology of these pathogens. Molecular tools that increase the accuracy and sensitivity of traditional methods have been developed to identify the hosts of potential insect vectors. However, to the present group's knowledge, molecular studies that identify the hosts of C. imicola in Europe are lacking. The present study genetically characterizes the barcoding region of C. imicola trapped on farms in southern Spain and identifies its vertebrate hosts in the area. The report also reviews available information on the blood-feeding patterns of C. imicola worldwide. Culicoides imicola from Spain feed on blood of six mammals that include species known to be hosts of the BT and AHS viruses. This study provides evidence of the importance of livestock as sources of bloodmeals for C. imicola and the relevance of this species in the transmission of BT and AHS viruses in Europe., This study was partially funded by projects CGL2015‐65055‐P from the Spanish Ministry of Economy and Competitiveness, P11‐RNM‐7038, from the Junta de Andalucía and a Microproyecto funded by the internal Estación Biológica de Doñana proposal financed by the Spanish Ministry of Economy and Competitiveness through the Severo Ochoa Centres of Excellence in Research, Development and Innovation programme (SEV‐2012‐0262). Funds were also partially provided by EU grant HEALTH.2010.2.3.3‐3 project 261391 EuroWestNile. JM‐dlP and MF were funded by a Juan de la Cierva contract and a Formación de Profesorado Universitario grant, respectively.

Equine encephalosis virus (EEV) is an orbivirus transmitted by Culicoides species. Most infected horses show mild clinical signs and mortality is usually very low. EEV is closely related and similarly transmitted to other, more pathogenic and economically important, orbiviruses such as African horse sickness virus (AHSV), bluetongue virus (BTV) and epizootic haemorrhagic disease viruses (EHDV), and may serve as an indicator for possible transmission of the latter. Israel has been reported to be endemic for EEV since 2001. This study was initiated to re‐evaluate the current seroprevalence and risk factors for EEV exposure in Israel, and to assess, for the first time, the seroprevalence of EEV in Palestine and Jordan. Three hundred and sixteen serum samples were collected from apparently healthy horses at 21 farms in Israel, 66 horses at nine farms in Palestine and 100 horses at three farms in Jordan. The presence of EEV antibodies was detected by a serum neutralization assay. Seroprevalence of EEV was 58.2% (184/316 horses) in Israel, 48.5% (32/66 horses) in Palestine and 2% (2/100 horses) in Jordan. Seroprevalence in Jordan was significantly lower than in Israel and Palestine (P

There is very little information available about Culicoides species (Diptera: Ceratopogonidae) in the western Brazilian Amazon. However, studies of the fauna of this region are essential to knowledge of the species and potential vectors within it. Thus, the present study aims to evaluate the abundance, richness and composition of Culicoides species in rural areas in the state of Rondonia, Brazil. Culicoides specimens were collected in forest and pasture environments in the municipality of Porto Velho, using light traps. A total of 1708 individuals (1136 females and 572 males) belonging to 33 species were collected; 28 of these samples represent new records for the state of Rondonia and include the first record of Culicoides contubernalis in Brazil. Culicoides insignis was the most abundant species (86.1%). Species richness was greater in forest areas (32 species, 96.96%), whereas pastures presented the greatest number of Culicoides captured (n = 1540, 90.1%). This study shows that Culicoides populations differ between forest and pasture environments and indicates that the abundance of C. insignis is an important factor in epidemiological vigilance studies in the region.

In the summer of 2014, in the central part of The Netherlands, Culicoides spp. (Diptera: Ceratopogonidae) attack rates, biting rates, and preferred landing sites were determined for a pair of Fjord horses maintained permanently at pasture in an area devoid of cattle. Eleven body regions of the horses were screened for midges, each region sampled randomly for 5 min using a handheld mouth aspirator (pooter). Observations were confined to the hour immediately before and after sunset. Culicoides spp. were obtained from every body region, of which the four most abundant species – Culicoides chiopterus (Meigen), Culicoides punctatus (Meigen), the species complex Culicoides obsoletus (Meigen), and Culicoides dewulfi Goetghebuer – all were proven or potential vectors for arboviral diseases in livestock. Culicoides spp. activity was distinctly bimodal across the day, surging at sunset and 1 h after sunrise. Midges were inactive between 11:00 and 16:00 hours, these hours marking the time of day when horses can be pastured most safely but, thereafter, to avoid escalating attacks, would have to be stabled protectively. Around sunset, the mean attack rate of the four most abundant species ranged from 3.0 to 11.7 midges per min; of these, C. dewulfi and C. chiopterus were reared out of the dung of experimental horses. The Netherlands is home to the world's densest horse population (11 per km2), of which half are estimated to stay outdoors permanently with no access to protective housing. In the absence of a preventive vaccination policy, it is difficult to envisage how horses in northern Europe will be protected from infection during an outbreak of a Culicoides-transmitted disease like African horse sickness.

Culicoides sonorensis Wirth & Jones (Diptera: Ceratopogonidae) is the primary North American vector of bluetongue virus (BTV), which can cause high morbidity and mortality in ruminant livestock or wildlife. Worldwide, most Culicoides surveillance relies on light (usually UV) traps typically placed near animals or larval development sites. However, the trapping method can cause sex, species and parity biases in collections. We collected C. sonorensis from three dairies in California using suction traps baited with CO2 , UV light or CO2 + UV placed near animals, wastewater ponds, or in fields. Higher numbers of parous females were collected using CO2 + UV traps, although this difference was only significant on one dairy. UV traps were poor at collecting nulliparous females, but the addition of UV to a trap increased the abundance of males in a collection. Traps set in open fields collected significantly higher numbers of males and females than in either of the other two locations. In some cases, there was a significant interaction between the trap type and site. We discuss the limitations of traditional trapping methodologies for C. sonorensis and make suggestions for vector surveillance.

Culicoides (Diptera: Ceratopogonidae) midges are the biological vectors of a number of arboviruses of veterinary importance. However, knowledge relating to the basic biology of some species, including their host-feeding preferences, is limited. Identification of host-feeding preferences in haematophagous insects can help to elucidate the transmission dynamics of the arboviruses they may transmit. In this study, a series of semi-quantitative real-time polymerase chain reaction (qPCR) assays to identify the vertebrate host sources of bloodmeals of Culicoides midges was developed. Two pan-reactive species group and seven species-specific qPCR assays were developed and evaluated. The assays are quick to perform and less expensive than nucleic acid sequencing of bloodmeals. Using these assays, it was possible to rapidly test nearly 700 blood-fed midges of various species from several geographic locations in Australia.

The mixing of an insecticide with sugar solution creates an oral toxin or insecticidal sugar bait (ISB) useful for reducing adult insect populations. The ability of ISBs to kill the biting midge Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae), a vector of bluetongue virus, epizootic hemorrhagic disease and vesicular stomatitis viruses, was tested. The commercial insecticide formulations (percentage active ingredient) tested included bifenthrin, cyfluthrin, deltamethrin, permethrin, dinotefuran, imidacloprid, thiamethoxam and spinosad. Mortality rates were determined for various concentrations of commercial formulations (0.01, 0.05, 0.1, 1, 2 and 3%) and observed at 1, 4, 10 and 24 h post-exposure to the ISB. In the first set of assays, laboratory-reared midges were fed sugar ad libitum and then exposed to insecticide-treated sugar solutions to measure mortality. The second assay assessed competitive feeding: midges were provided with a control sugar solution (10% sucrose) in one vial, and a sugar and insecticide solution in another. Pyrethroid treatments resulted in the greatest mortality in the first hour at the lowest concentrations and spinosad consumption resulted in the least mortality. Biting midges were not deterred from feeding on the 1% ISB solutions despite the presence of an insecticide-free alternative source of sugar.

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) may transmit several arboviruses to ruminant livestock. The species of the Obsoletus group are considered to be among the most important vectors of bluetongue virus (BTV) in northern Europe. As agricultural environments offer suitable habitats for the development of their immature stages, the emergence of adult Culicoides from potential breeding sites was investigated at 20 cattle farms throughout Germany in 2012 and 2013. In analyses of species-specific habitat preferences and relationships between Culicoides abundance in breeding substrates and their physicochemical characteristics, dungheaps emerged as the most important substrate for the development of Culicoides obsoletus sensu stricto (s.s.) (Meigen), whereas Culicoides chiopterus (Meigen) and Culicoides dewulfi Goetghebuer were generally restricted to cowpats. A decreasing pH value was associated with a higher abundance or a higher probability of observing these three species. Furthermore, the abundance of C. obsoletus s.s. was positively related to increasing moisture. Dungheaps were very productive breeding sites for this species and are therefore suggested as a target for potential control measures.

The biting midge Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae) transmits pathogens to both livestock and wildlife. Biting midge surveillance relies heavily on light traps for collection; however, little is known about the light spectra preferences of C. sonorensis midges. A light assay arena was constructed and light-emitting diodes (LEDs) of various light spectra were used as light sources to evaluate midge photoattraction. A comparison of responses to light spectra indicated the highest proportions of C. sonorensis were attracted to ultraviolet (UV) light and that midges differentiated 10-nm differences in wavelength. Stronger intensities of UV light resulted in greater attraction. Midges exhibited both sugar-seeking and escape behaviours under different conditions of sugar supplementation before and during the experiment. These behaviours occurred with lights of 355 nm and 365 nm in wavelength. Based on the results of this study, the attraction of C. sonorensis to light traps can be improved through the use of bright LEDs at 355 nm or 365 nm.

The light trap is the tool of choice for conducting large-scale Culicoides (Diptera: Ceratopogonidae) vector surveillance programmes. Its efficacy is in doubt, however. To assess this, hourly changes in Culicoides activity over the 24-h diel were determined comparatively by way of light trapping and aerial sweeping, and correlated against light intensity. In the Netherlands, sweeping around cattle at pasture revealed that, in early summer, Culicoides are active throughout the diel, and that their abundance peaks during the crepuscular period and falls to a low during the brightest hours of the day. By contrast, the light trap was able to accumulate Culicoides only at night (i.e. after illuminance levels had dropped to 0 lux and midge activity had begun to decline). Although Culicoides chiopterus and species of the Culicoides obsoletus complex were similarly abundant around livestock, they differed critically in their hours of peak activity, being largely diurnal and nocturnal, respectively. This polarity helps to explain why, routinely, the C. obsoletus complex dominates light trap collections and C. chiopterus does not. Inability to accumulate Culicoides at light intensity levels above 0 lux means that, at ever-higher latitudes, particularly beyond 45° N, the progressive northward lengthening of the twilight period will have an increasingly adverse impact upon the efficacy of the light trap as a vector surveillance tool.

To implement risk management against diseases transmitted by species of Culicoides Latreille, 1809 (Diptera: Ceratopogonidae), it is essential to identify all potential vectors. Light traps are the most commonly used tool for the collection of Culicoides midges. Given the indiscriminate artificial attraction of light, traps will collect all night-flying insects rather than only livestock-associated Culicoides midges. Factors that may increase the efficacy of traps, especially for livestock-associated Culicoides midges, require investigation. In the present study, results obtained with Centers for Disease Control (CDC) and Onderstepoort light traps baited with carbon dioxide (CO2 ) were compared with those of unbaited controls. Comparisons were made using two replicates of a 4 × 4 randomized Latin square design. With both trap types, the mean numbers of Culicoides midges collected in 16 baited traps were higher than those caught in 16 unbaited traps. Although exceptionally low numbers were collected with the CDC traps, the increases in the numbers and frequency of collection of Culicoides imicola Kieffer, 1913 were more pronounced in the CDC traps compared with the Onderstepoort traps. These results indicate that the addition of CO2 may increase the efficiency of these traps for the collection of C. imicola and other livestock-associated Culicoides species.

Bluetongue virus (BTV) is the type species of the genus Orbivirus within the family Reoviridae (the largest double-stranded ribonucleic acid (dsRNA) virus family). BTV is an arthropod-borne pathogen that is transmitted by the bites of its vector, the Culicoides biting midges, which transmit the virus between its ruminant hosts. The virus can infect most species of domestic and wild ruminants. BT clinical signs are most common in sheep and some species of deer. In recent years, the distribution of BTV has changed considerably, particularly in Europe. New outbreaks have occurred each year from 1998 in southern and central Europe, involving several strains from 10 different serotypes. In 2006, BTV serotype 8 caused an outbreak approximately 5° further north in Europe than ever before, infecting ruminants in a wide area across northern Europe. A new virus (Toggenburg virus) has been identified in goats from Switzerland in 2008 and designated as the twenty-fifth serotype of BTV and another one isolated from goats in Corsica in France more recently and was designated as serotype 27. Additional serotypes have been detected/isolated, bringing now the number of serotypes to 29. Key Concepts Genus Orbivirus is the most diverse genus of family Reoviridae containing 32 virus species. Bluetongue disease affects mainly sheep causing severe clinical signs and high mortality. Bluetongue virus is transmitted by competent species of Culicoides midges. Phylogenetic analysis identifies two major groups of orbiviruses, those with their subcore shell protein (T2 protein) being the VP2 and those with the T2 being VP3. Orbiviruses are nonturreted members of family Reoviridae, and the atomic structure of BTV core has been determined. Keywords: orbivirus; Reoviridae; dsRNA virus; BTV; bluetongue; Culicoides

In 2011, Culicoides (Diptera: Ceratopogonidae) were collected at 16 locations covering four regions of Belgium with Onderstepoort Veterinary Institute (OVI) traps and at two locations with Rothamsted suction traps (RSTs). Quantification of the collections and morphological identification showed important variations in abundance and species diversity between individual collection sites, even for sites located in the same region. However, consistently higher numbers of Culicoides midges were collected at some sites compared with others. When species abundance and diversity were analysed at regional level, between-site variation disappeared. Overall, species belonging to the subgenus Avaritia together with Culicoides pulicaris (subgenus Culicoides) were the most abundant, accounting for 80% and 96% of all midges collected with RSTs and OVI traps, respectively. Culicoides were present during most of the year, with Culicoides obsoletus complex midges found from 9 February until 27 December. Real-time reverse-transcription polymerase chain reaction screening for Schmallenberg virus in the heads of collected midges resulted in the first detection of the virus in August 2011 and identified C. obsoletus complex, Culicoides chiopterus and Culicoides dewulfi midges as putative vector species. At Libramont in the south of Belgium, no positive pools were identified.

Many biting midges of the genus Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) are competent vectors of a diverse number of pathogens. The identification of their feeding behaviour and of vector–host associations is essential for understanding their transmission capacity. By applying two different nested polymerase chain reaction (PCR) assays, of which one targeted the avian cyt b gene and the other targeted the COI gene of a wide range of vertebrates, we identified the blood hosts of six biting midge species including Culicoides circumscriptus, Culicoides festivipennis, Culicoides punctatus, Culicoides pictipennis, Culicoides alazanicus and Culicoides cf. griseidorsum, the latter two of which are reported in Bulgaria for the first time. Bird DNA was found in 50.6% of 95 investigated bloodmeals, whereas mammalian DNA was identified in 13.7%. Two Culicoides species were found to feed on both birds and mammals. There was remarkable diversity in the range of avian hosts: 23 species from four orders were identified in the abdomens of four Culicoides species. The most common bird species identified was the magpie, Pica pica (n = 7), which was registered in all four ornithophilic biting midge species. Six bloodmeals from the great tit, Parus major, were recorded only in C. alazanicus. None of the studied species of Culicoides appeared to be restricted to a single avian host.

This study examined environmental and geographical factors affecting the distribution of the bluetongue virus vectors Culicoides brevitarsis Kieffer, C. actoni Smith, C. fulvus Sen and Das Gupta and C. wadai Kitaoka in Australia using graphical analysis. Calculations and mapping were carried out in the R programming environment using freely available data. Average low temperatures of 13.5°C in June and in September were estimated as thresholds and expressed as contours at which C. brevitarsis activity ceases and then recommences during and after winter, respectively. The June threshold also designated areas where populations would be unable to survive the winter. There was substantial variability in the contours between years, around which the activity of C. brevitarsis would be correspondingly dynamic. Culicoides wadai, C. actoni and C. fulvus appeared to be less tolerant of low temperatures. A high temperature limit was proposed at a maximum 35°C during summer. Culicoides brevitarsis was found in areas above and below this temperature, while C. actoni, C. fulvus and C. wadai occurred mainly in coastal environments at maximum temperatures of less than 35°C. Possible effects of global warming were considered with 1°C or 2°C rises in temperature. Changes to low temperature thresholds could expose greater areas of southern Australia to vector activity, while temperature rises in the north could increase the size of areas in which temperatures are lethal to the vectors. Rainfall had no apparent effect on C. brevitarsis during the northern wet and dry seasons. Distributions of C. brevitarsis and cattle overlapped until both ended with the climatic change to desert, which formed a physical barrier preventing C. brevitarsis movement to the south. Temperature restricted associations between C. brevitarsis and sheep. The other Culicoides spp. were found outside major cattle and sheep production areas. Apart from seasonal and dynamic activity around the contours, cattle and sheep in areas below a June average temperature of 13.5°C should remain vector free at current climatic temperatures.

Light trap surveillance across northern Australia and Papua New Guinea (PNG) has detected the presence of several Oriental species of Culicoides not previously reported from those countries and which appear to have arrived in recent times. Detections of C. nudipalpis Delfinado in Western Australia, C. flavipunctatus Kitaoka and C. palpifer Das Gupta and Ghosh in the Northern Territory and of C. flavipunctatus, C. fulvus Sen and Das Gupta and C. orientalis Macfie in Queensland (Qld) provide evidence of multiple pathways for incursions of biting midges into northern Australia. Of these, only C. fulvus appears to have established. Additionally, three species, C. fulvus, C. wadai Kitaoka and C. brevipalpis Delfinado, are newly reported from PNG and all appear to be well established. The arrival in PNG of C. fulvus and C. brevipalpis, both not previously reported from Qld, suggests that pathways exist for the entry of Oriental insects into New Guinea directly from Asia, rather than via Australia. Molecular analyses using DNA barcodes (partial mitochondrial cytochrome oxidase subunit one sequences) confirmed morphological identification of specimens and additionally provided strong evidence relating to the source of these incursions. At least two of these species are vectors of important livestock viruses and are likely to impact on the epidemiology of these viruses as they continue to disperse.

SCHMALLENBERG virus (SBV) emerged during the summer of 2011 in north-western Europe and was identified for the first time in dairy cattle herds in Germany (Hoffmann and others 2012). The virus belongs to the Simbu serogroup of Orthobunyaviruses and many similarities have been reported with the closely related Akabane virus regarding its transmission, pathogenesis and induced clinical signs. SBV causes mild clinical signs in adult ruminants, ranging from fever, diarrhoea and temporarily reduced milk production. However, infection of pregnant cattle, sheep and goats can cause transplacental transmission leading months later to abortions, stillbirths and congenital malformations characterised by the arthrogryposis-hydranencephaly syndrome in newborns (Doceul and others 2013). After its first identification in Germany in 2011, SBV spread rapidly and widely over a large part of Europe, including the UK (EFSA 2014). Indigenous European Culicoides biting midges, which were already known to have spread bluetongue virus serotype 8 after its emergence in Europe in 2006 (Mehlhorn and others 2007, Meiswinkel and others 2007), were found to be responsible for the rapid transmission of SBV (De Regge and others 2012, 2015). Seroprevalence studies showed that the majority of domestic ruminants became infected in SBV-affected countries (Meroc and others 2013, 2014, Wernike and others 2014). In 2012, SBV spread further over Europe and evidence for renewed but lower levels of SBV circulation in countries affected in 2011 was found (Conraths and others 2013, Afonso and others 2014, De Regge and others 2014). From spring 2013 onwards, the number …

Several species of Culicoides (Diptera: Ceratopogonidae) are vectors of pathogens, such as the bluetongue (BTV) and Schmallenberg (SBV) viruses, which cause important diseases in domestic and wild ruminants. As wild ruminants can contribute to overwintering and epizootics of both diseases, knowledge of the host-feeding behaviour of Culicoides in natural ecosystems is important to better understand their epidemiology. Blood-engorged Culicoides females trapped in natural areas inhabited by different wild ruminant species were genetically analysed to identify host species. The origin of bloodmeals was identified in 114 females of 14 species of Culicoides. A total of 104 (91.1%) Culicoides fed on mammals and 10 (8.9%) on birds. The most abundant host identified was red deer (66.7%), followed by humans (13%) and fallow deer (6.1%). Eleven of the 14 species of Culicoides fed exclusively on mammalian hosts. Among them, five are mammalophilic species considered to be important BTV and/or SBV vectors. The results of the present study confirm that Culicoides imicola, Culicoides obsoletus, Culicoides scoticus, Culicoides pulicaris and Culicoides punctatus fed on wild ruminants, and therefore support the hypothesis that these species can act as bridge vectors by facilitating the circulation of pathogens between wild and domestic ruminant communities. info:eu-repo/semantics/acceptedVersion

The performance of different bluetongue control measures related to both vaccination and protection from bluetongue virus (BTV) vectors was assessed. By means of a mathematical model, it was concluded that when vaccination is applied on 95% of animals even for 3 years, bluetongue cannot be eradicated and is able to re‐emerge. Only after 5 years of vaccination, the infection may be close to the eradication levels. In the absence of vaccination, the disease can persist for several years, reaching an endemic condition with low level of prevalence of infection. Among the mechanisms for bluetongue persistence, the persistence in the wildlife, the transplacental transmission in the host, the duration of viraemia and the possible vertical transmission in vectors were assessed. The criteria of the current surveillance scheme in place in the EU for demonstration of the virus absence need revision, because it was highlighted that under the current surveillance policy bluetongue circulation might occur undetected. For the safe movement of animals, newborn ruminants from vaccinated mothers with neutralising antibodies can be considered protected against infection, although a protective titre threshold cannot be identified. The presence of colostral antibodies interferes with the vaccine immunisation in the newborn for more than 3 months after birth, whereas the minimum time after vaccination of animal to be considered immune can be up to 48 days. The knowledge about vectors ecology, mechanisms of over‐wintering and criteria for the seasonally vector‐free period was updated. Some Culicoides species are active throughout the year and an absolute vector‐free period may not exist at least in some areas in Europe. To date, there is no evidence that the use of insecticides and repellents reduce the transmission of BTV in the field, although this may reduce host/vector contact. By only using pour‐on insecticides, protection of animals is lower than the one provided by vector‐proof establishments., This publication is linked to the following EFSA Supporting Publications article: http://onlinelibrary.wiley.com/doi/10.2903/sp.efsa.2017.EN-1182/full, http://onlinelibrary.wiley.com/doi/10.2903/sp.efsa.2017.EN-1171/full

Horses are affected by a wide variety of arthropod ectoparasites, ranging from lice which spend their entire life on the host, through ticks which feed over a period of days, to numerous biting insects that only transiently visit the host to feed. The presence of ectoparasites elicits a number of host responses including innate inflammatory responses, adaptive immune reactions and altered behaviour; all of which can reduce the severity of the parasite burden. All of these different responses are linked through immune mechanisms mediated by mast cells and IgE antibodies which have an important role in host resistance to ectoparasites, yet immune responses also cause severe pathological reactions. One of the best described examples of such pathological sequelae is insect bite hypersensitivity (IBH) of horses; an IgE-mediated type 1 hypersensitivity to the salivary proteins of Culicoides spp. associated with T-helper-2 production of IL4 and IL13. Importantly, all horses exposed to Culicoides have an expanded population of Culicoides antigen-specific T cells with this pattern of cytokine production, but in those which remain healthy, the inflammatory reaction is tempered by the presence of FoxP3+ CD4+ regulatory T cells that express IL10 and TGF-beta, which suppresses the IL4 production by Culicoides antigen-activated T cells.

Most haematophagous insect vectors can also use sugar as an energy source; thus their sugar-feeding behaviour influences their longevity and blood-feeding rate and hence their vectorial capacity. Scant information is available on the sugar-feeding behaviour of Culicoides Latreille biting midges (Diptera: Ceratopogonidae), which are vectors of bluetongue and Schmallenberg viruses. The longevity of laboratory-reared Culicoides nubeculosus (Meigen) under fluctuating temperatures (16 and 28 °C) and with access to water or water and blood was on average 6.4 days and 8.9 days, respectively, which was around one third of the lifespan of siblings with access to sugar or sugar and blood (22.2 days and 27.1 days, respectively). Access to honeydew significantly increased the midge's longevity, whereas the provision of extrafloral nectaries had no impact. Females with access to sugar produced a significantly higher number of eggs (65.5 ± 5.2) than their starved sisters (45.4 ± 8.4). More than 80% of field-caught female Culicoides from the two most abundant European groups, Obsoletus (n = 2243) and Pulicaris (n = 805), were fructose-positive. Fructose-positivity was high in all physiological stages and no seasonal variability was noted. The high rate of natural sugar feeding of Culicoides offers opportunities for the development of novel control strategies using toxic sugar baits and for the monitoring of vector-borne diseases using sugar-treated FTA (nucleic acid preservation) cards in the field.

The efficacy of 23 compounds in repelling Culicoides biting midges (Diptera: Ceratopogonidae), particularly Culicoides obsoletus (Meigen) females, was determined by means of a Y-tube olfactometer. The 10 most effective compounds were further evaluated in landing bioassays. The six most promising compounds (including chemical and plant-derived repellents) were evaluated at 10% and 25% concentrations in field assays using Centers for Disease Control (CDC) light traps. At least three compounds showed promising results against Culicoides biting midges with the methodologies used. Whereas olfactometer assays indicated DEET at 1 µg/µL to be the most effective repellent, filter paper landing bioassays showed plant-derived oils to be better. Light traps fitted with polyester mesh impregnated with a mixture of octanoic, decanoic and nonanoic fatty acids at 10% and 25% concentrations collected 2.2 and 3.6 times fewer midges than control traps and were as effective as DEET, which is presently considered the reference standard insect repellent. The best plant-derived product was lemon eucalyptus oil. Although these have been reported as safe potential repellents, the present results indicate DEET and the mixture of organic fatty acids to be superior and longer lasting.

An illustrated key to aid in separating adult females of the 64 species of Culicoides reported from the Northern Territory, Western Australia and South Australia, and some important livestock pest species not present in these regions is presented. The key uses characters visible under a stereomicroscope and does not attempt to distinguish those species that are separable only on characters visible on slide-mounted specimens. C. asiana Bellis is proposed as a replacement name for C. asiatica Bellis.

Culicoides Latreille (Diptera: Ceratopogonidae) are vectors of several arboviruses, including bluetongue virus (BTV) and African horse sickness virus (AHSV), which cause diseases in, respectively, sheep and cattle, and horses, and have economic repercussions mainly as a result of trade restrictions. Insecticides can be used to reduce vector populations and hence the spread of disease. Despite the economic importance of these diseases, relatively few studies have evaluated the efficacy of commercially available insecticides and the effectiveness of treated nets against Culicoides species. The aim of the present study was to evaluate the insecticidal effect of commercially available polyethylene nets (ZeroVector(®) ) treated with deltamethrin (4.4 g/kg ± 15%) on Culicoides species. Laboratory and field trials were conducted in Culicoides populations collected in Majorca in the Balearic Islands, Spain. The present study shows that deltamethrin-treated nets provoke high and rapid mortality (90-100%) in Culicoides midges under laboratory conditions and increase mortality by 13% when deployed in the field.