Spatial and temporal distribution of bluetongue and its Culicoides vectors in Bulgaria

Surveillance of Culicoides (Diptera: Ceratopogonidae) biting midges was carried out between 2001 and 2003, at 119 sites within a 50 x 50-km grid distributed across Bulgaria, using light trap collections around the time of peak adult midge abundance. Sentinel and ad hoc serum surveillance of hosts susceptible to bluetongue infection was carried out at around 300 sites between 1999 and 2003. Following the initial incursion of bluetongue virus 9 (BTV-9) into Bourgas province in 1999, affecting 85 villages along the southern border, a further 76 villages were affected along the western border in 2001, with outbreaks extending as far north as 43.6 degrees N. The BTV-9 strain in circulation was found to have a low pathogenicity for Bulgarian sheep populations, with less than 2% of susceptible individuals becoming sick and seroconversions detected up to 30 km from recorded outbreaks in the south. The major Old World vector Culicoides imicola Kieffer was not detected among over 70,000 Culicoides identified in summer collections, suggesting that BTV-9 transmission in Bulgaria was primarily carried out by indigenous European vectors. The most likely candidates, the Palaearctic species complexes - the Culicoides obsoletus Meigen and C. pulicaris L. complexes - were widespread and abundant across the whole country. The C. obsoletus complex represented 75% of all individuals trapped in summer and occurred in high catch sizes (up to 15,000 individuals per night) but was not found across all outbreak sites, indicating that both Palearctic complexes probably played a role in transmission. Within the C. pulicaris complex, only C. pulicaris s.s., C. punctatus Meigen and C. newsteadi Austen were sufficiently abundant and prevalent to have been widely involved in transmission, whilst within the C. obsoletus complex most trapped males were C. obsoletus s.s. Adult vectors were found to be largely absent from sites in west Bulgaria for a period of at least 3 months over winter, which, taken along with the spatiotemporal pattern of outbreaks in the region between years, indicates the virus may be overwintering here by an alternative mechanism - either by covert persistence in the vertebrate host or possibly by persistence in larval stages of the vector.     

Spatial distribution of Culicoides species in Portugal in relation to the transmission of African horse sickness and bluetongue viruses

Surveillance of Culicoides (Diptera: Ceratopogonidae) biting midge vectors was carried out at 87 sites within a 50 x 50 km grid distributed across Portugal, using light trap collections at the time of peak midge abundance. Culicoides imicola (Kieffer) made up 66% of the 55 937 Culicoides in these summer collections. It was highly abundant in the central eastern portion of Portugal, between 37 degrees 5' N and 41 degrees 5' N, and in a band across to the Lisbon peninsula (at around 38 degrees 5' N). Of all the complexes, its distribution was most consistent with that of previous outbreaks of Culicoides-borne disease, suggesting that it may remain the major vector in Portugal. Its distribution was also broadly consistent with that predicted by a recent climate-driven model validating the use of remote sensing datasets for modelling of Culicoides distribution. Adult C. imicola were found to have overwintered at 12 of 20 sites re-surveyed in winter but it did so in very low numbers. Culicoides obsoletus (Meigen) and Culicoides pulicaris (Linnaeus) complex midges were widespread despite their low summer abundance. The observed coincidence of high abundances of C. imicola and high abundances of C. pulicaris in summer lead us to suggest that C. imicola could bring African horse sickness virus or bluetongue virus into contact with C. pulicaris and the latter complex, together with C. obsoletus, could then transmit these viruses across much wider areas of Europe. The fact that adult C. pulicaris are present in high abundances in winter may provide a mechanism by which these viruses can overwinter in these areas.     

Spatial distribution of bluetongue virus and its Culicoides vectors in Sicily

During the recent Mediterranean epizootic of bluetongue, an extensive programme of serological and vector (Culicoides biting midges (Diptera: Ceratopogonidae)) surveillance was carried out across Sicily. This paper presents the analysis of 911 light trap catches collected at the times of peak Culicoides abundance (summer to autumn 2000-2002) in 269 sites, in order to produce detailed maps of the spatial distribution of the main European vector, Culicoides imicola Kieffer and that of potential novel vectors. Whereas C. imicola was found at only 12% of sites, potential novel vectors, Culicoides obsoletus group Meigen, Culicoides pulicaris Linnaeus and Culicoides newsteadi Austen were present at over 50% of sites. However, the spatial distribution of C. imicola showed the closest correspondence to that of the 2000 and 2001 bluetongue (BT) outbreaks and its presence and abundance were significant predictors of the probability of an outbreak, suggesting that it was the main vector during these years. Although C. imicola may have played a role in transmission in several sites near Paternó, it was absent from the majority of sites at which outbreaks occurred in 2002 and from all sites in the province of Messina. All three potential novel vectors were widespread across sites at which outbreaks occurred during 2002. Of these, C. newsteadi was an unlikely candidate, as it was significantly less prevalent in outbreak vs. non-outbreak sites in Messina. It is hypothesized that the yearly distribution and intensity of outbreaks is directly attributable to the distribution and abundance of the vectors involved in transmission during each year. When C. imicola operated as the main vector in 2000 and 2001, outbreaks were few in number and were restricted to coastal regions due to low abundance and prevalence of this species. In 2002, it is hypothesized that BTV transmission was handed over to more prevalent and abundant novel vector species, leading to numerous and widespread outbreaks and probably to overwintering of the virus between 2001 and 2002. Based on catch ranges in outbreak vs. non-outbreak sites, it is tentatively suggested that nightly catches of 400 or more C. obsoletus and 150 or more C. pulicaris allow BTV transmission at a site, and provide a strategy for a fuller examination of the relationship between BTV transmission and the abundance and distribution of different vector species.     

五种库蠓脂肪酸的研究

本文用气相色语法分析了不显库蠓(Culicoides obsoletus)、兴安库蠓(C, sinanoensis)、怒江库蠓(C.Nujiangensis)、灰黑库蠓(C.Pulicaris)和刺整库蠓(C.Punctatus)等5种库蠓雌虫脂 肪醛的组成和含量。 结果表明,5种库蠓均含有12-14种主要脂肪酸,并以棕榈油酸(C_16：1)、棕搁酸 (C_16：0)、油酸(C_18：1)、硬脂酸(C_18：0)含量最高,共占脂肪酸总量的70%以上。对主要脂肪酸的含量进行聚类分析,发现不显库蠓种团内任意两种库蠓间的欧氏距离小于不显库蠓种团种类与灰黑库蠓种团种类间的欧氏距离,说明脂肪酸的定性、定量分析结果对库蠓分类有一定的参考价值。     

Modelling the distributions of Culicoides bluetongue virus vectors in Sicily in relation to satellite-derived climate variables

Surveillance data from 268 sites in Sicily are used to develop climatic models for prediction of the distribution of the main European bluetongue virus (BTV) vector Culicoides imicola Kieffer (Diptera: Ceratopogonidae) and of potential novel vectors, Culicoides pulicaris Linnaeus, Culicoides obsoletus group Meigen and Culicoides newsteadi Austen. The models containing the 'best' climatic predictors of distribution for each species, were selected from combinations of 40 temporally Fourier-processed remotely sensed variables and altitude at a 1 km spatial resolution using discriminant analysis. Kappa values of around 0.6 for all species models indicated substantial levels of agreement between model predictions and observed data. Whilst the distributions of C. obsoletus group and C. newsteadi were predicted by temperature variables, those of C. pulicaris and C. imicola were determined mainly by normalized difference vegetation index (NDVI), a variable correlated with soil moisture and vegetation biomass and productivity. These models were used to predict species presence in unsampled pixels across Italy and for C. imicola across Europe and North Africa. The predicted continuous presence of C. pulicaris along the appenine mountains, from north to south Italy, suggests BTV transmission may be possible in a large proportion of this region and that seasonal transhumance (seasonal movement of livestock between upland and lowland pastures) even in C. imicola-free areas should not generally be considered safe. The predicted distribution of C. imicola distribution shows substantial agreement with observed surveillance data from Greece and Iberia (including the Balearics) and parts of mainland Italy (Lazio, Tuscany and areas of the Ionian coast) but is generally much more restricted than the observed distribution (in Sardinia, Corsica and Morocco). The low number of presence sites for C. imicola in Sicily meant that only a restricted range of potential C. imicola habitats were included in the training set and that predictions could only be made within this range. Future modelling exercises will use abundance data collected according to a standardized protocol across the Mediterranean and, for Sicily in particular, should include non-climatic environmental variables that may influence breeding site suitability such as soil type.     

Light and suction trap catches of Culicoides midges in southern England

1. The seasonal flight periods are given for eleven species or species groups of Culicoides midges at Pirbright, Surrey, U.K., from 1979 to 1981 and in 1983. 2. Most species were relatively uniformly distributed both within the study site and between years; only C.clintoni Boorman was limited in its spatial distribution and only C.segnis Campbell and Pelham-Clinton varied greatly in numbers between years. 3. Species fall into one of three patterns of seasonal abundance: spring emergers, summer emergers, or spring to autumn emergers. 4. Only the C.obsoletus group showed a pattern of emergence that could correspond to distinct generations, of one or more species in the group. Three distinct emergences were observed, and possible explanations for these discussed. 5. For all species except C.impunctatus Goetghebuer the catches from suction/light and suction only traps are poorly correlated whereas for C.impunctatus they are highly correlated. This may suggest that this species is not greatly attracted to light.     

Culicoides midge trap enhancement with animal odour baits in Scotland

Examples of the commercial trap Mosquito Magnet Pro (MMP emitting attractant 1-octen-3-ol in carbon dioxide 500 mL/min generated from propane fuel), were run 24 h/day on the Isle of Skye, Scotland, during June-August 2001 and evaluated for catching Culicoides biting midges (Diptera: Ceratopogonidae). From 30 days trapping, the catch averaged 2626 +/- 1358 Culicoides females/trap/day (mean +/- SE, range 558 +/- 139 to 6088 +/- 3597, for five sets of six consecutive nights), predominantly the pest Culicoides impunctatus Goetghebuer (68% overall), plus C. vexans (Staeger) > C. delta Edwards > C. pulicaris (L.) > C. lupicaris Downs & Kettle > C. albicans (Winnertz) > other Culicoides spp. Attempts were made to enhance the odour baiting system by adding hexane-extracts (2.1 mg/day) of hair samples from large host animals, resulting in the following effects on Culicoides collections: sheep - 53 %, red deer - 26 %, calf + 20%, pony + 40%, water buffalo + 262%, with greatest increases for C. impunctatus and C. pulicaris. Serial concentrations of these animal extracts (10(-1) - 10(-3) x 2.2 g/mL) were assayed on parous female C. impunctatus response in a Y-tube olfactometer (air-flow 150 mL/min), and by electroantennogram (EAG) on Culicoides nubeculosus Meigen laboratory-reared parous females. Positive behavioural responses to host odours were dose-dependent: the water buffalo extract being most active (threshold 0.22 g/mL), similar to deer, whereas other host extracts were > or = 10-fold less active. Correspondingly, the EAG threshold was lowest for water buffalo, 10-fold greater for deer, calf and pony, but not detected for sheep. If the active component(s) of these host extracts can be identified and synthesized, they might be employed to improve the capture of Culicoides midges for local control by removal trapping.     

Phylogenetic analysis of Culicoides species from France based on nuclear ITS1-rDNA sequences

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) play important roles in the transmission of viral diseases affecting wild and domestic ruminants and horses, including Bluetongue (BT) and African horse sickness (AHS) respectively. In southern Europe, BT has been largely transmitted by the classical Afro-Asian vector Culicoides imicola Kieffer. However, other species such as C. obsoletus Meigen, C. scoticus Downs & Kettle and C. pulicaris Linné may also be involved in BTV transmission. As a consequence of the discovery of C. imicola followed by BTV-2 outbreaks on the island of Corsica in October 2000, further studies on these biting midges have been carried out. To better characterize the evolution and phylogenetic relations of Culicoides, molecular analysis in parallel with a morphology-based taxonomic approach were performed. Phylogenetic analyses of French Culicoides species were undertaken using the ribosomal DNA (rDNA) internal transcribed spacer 1 (ITS1) as a molecular target. This region was shown to be useful in understanding evolutionary and genetic relationships between species. Construction of several trees showed that molecular phylogeny within the genus Culicoides correlates not only with morphological-based taxonomy but also with ecological patterns.     

Evaluation of housing as a means to protect cattle from Culicoides biting midges, the vectors of bluetongue virus

The housing of animals at night was investigated as a possible means of protecting them from attack by Culicoides biting midges (Diptera: Ceratopogonidae), the vectors of bluetongue. Light-trap catches of Culicoides were compared inside and outside animal housing, in the presence and absence of cattle. A three-replicate, 4 × 4 Latin square design was used at four farms in Bala, north Wales, over 12 nights in May and June 2007, and the experiment repeated in October. In the two studies, respectively, >70 000 and >4500 Culicoides were trapped, of which 93% and 86%, respectively, were of the Culicoides obsoletus group. Across the four farms, in May and June, the presence of cattle increased catches of C. obsoletus by 2.3 times, and outside traps caught 6.5 times more insects than inside traps. Similar patterns were apparent in October, but the difference between inside and outside catches was reduced. Catches were strongly correlated with minimum temperature and maximum wind speed and these two variables explained a large amount of night-to-night variation in catch. Outside catches were reduced, to a greater extent than inside catches, by colder minimum temperatures and higher maximum wind speeds. These conditions occur more frequently in October than in May and June, thereby suppressing outside catches more than inside catches, and reducing the apparent degree of exophily of C. obsoletus in autumn. The results suggest that the risk of animals receiving bites from C. obsoletus is reduced by housing at both times of year and the benefit would be greatest on warm, still nights when outside catches are at their greatest.     

库蠓亚属6种库蠓翅形变化分析及亲缘关系探讨

本研究基于几何形态学测量分析方法对库蠓亚属6种库蠓(渐灰库蠓Culicoides grisescens、光胸库蠓Culicoides impunctatus、日本库蠓Culicoides nipponensis、灰黑库蠓Culicoides pulicaris、刺螫库蠓Culicoides punctatus、天祥库蠓Culicoides tienhsiangensis)的翅脉形态进行定量分析,以探讨6种库蠓种间的亲缘关系。首先对库蠓亚属6种、24头标本翅脉使用几何形态学标点法标记12个地标点,并通过普氏叠加、典型变量分析法、聚类分析法等对6种库蠓翅进行比较分析以及种间亲缘关系探讨。结果显示,翅形变化与亲缘关系结果一致,即6种库蠓的翅形均有显著差异(P<0.05),其中光胸库蠓和渐灰库蠓的翅形变化最小,亲缘关系最近,而日本库蠓与其他5种库蠓的翅形变化最大,亲缘关系最远。初步证实了几何形态学能够做为一种辅助工具对蠓科昆虫进行种间关系及分类研究。     

Phylogeny of the subgenus Culicoides and related species in Italy, inferred from internal transcribed spacer 2 ribosomal DNA sequences

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) include vectors for the economically important animal diseases, bluetongue (BT) and African horse sickness (AHS). In the Mediterranean Basin, these diseases are transmitted by four species of Culicoides: the first three belong in the subgenus Avaritia Fox and are Culicoides imicola Kieffer, Culicoides obsoletus (Meigen) and Culicoides scoticus Downes and Kettle; the fourth is Culicoides pulicaris (Linnaeus) in the subgenus Culicoides Latreille. In the Palaearctic Region, this subgenus (usually referred to as the C. pulicaris group) now includes a loose miscellany of some 50 taxa. The lack of clarity surrounding its taxonomy stimulated the present morphological and molecular study of 11 species collected in Italy. Phylogenetic analysis of nuclear ribosomal DNA internal transcribed spacer 2 (ITS2) sequence variation demonstrated a high degree of divergence. These results, combined with those from a parallel morphological study, disclosed: (1) that some previously described taxa should be resurrected from synonymy; (2) that there are new species to be described; (3) that the subgenus Culicoides (as currently employed) is a polyphyletic assemblage of four lineages - the subgenus Culicoides sensu stricto, the subgenus Silvicola Mirzaeva and Isaev, the subgenus Hoffmania Fox and the hitherto unrecognized Fagineus species complex. Each is discussed briefly (but not defined) and its constituent Palaearctic taxa listed. Strong congruence between morphological and molecular data holds promise for resolving many of the difficult taxonomic issues plaguing the accurate identification of vector Culicoides around the world.     

Toward development of neem-based repellents against the Scottish Highland biting midge Culicoides impunctatus

Oil of neem, from the tree Azadirachta indica A. Juss (Meliaceae), was evaluated for repellent and antifeedant activity against Culicoides biting midges (Diptera: Ceratopogonidae), by three complementary methods with serial dilutions. Electroantennograms revealed the sensitivity of Culicoides nubeculosus (Meigen) females to neem > or = 0.10%. Culicoides impunctatus Goetghebuer females were repelled by > or = 1% in a Y-tube olfactometer, Using a membrane feeder for wild-caught parous females of C. impunctatus, the proportion blood-feeding was significantly reduced by topical applications of neem oil > or = 0.10% concentrations, with blood-feeding completely prevented by > or =1%. On the basis of these response data, we developed 2% neem-based formulations for personal protection against biting midges.     

东北边境地带吸血蠓的区系和生态研究

我国东北吉林省和黑龙江省的边境地带共有18个县区,东起于吉林省珲春县,北止于我国最北端的黑龙江省漠河县,全长约2500km,位于东经122°4′—129°50′,北纬53°—41°25′之间,一般海拔500—1000m,年平均降水量300—700mm,昆虫地理区划属东北区大兴安岭亚区和长白山亚区。多年来,吸血蠓对边境     

Blood-sucking biting midges (Ceratopogonidae) in north central Siberia

6000 adult punkies were collected from man and animals over a period of 15 years in the Taimyr and Evenki Autonomous Okrugs of Krasnoyarsk Territory. Four species Culicoides chiopterus, C. pulicaris, C. fascipennis, C. alatavicus are reported from tundra and forest-tundra of Taimyr (68 degrees 00'-73 degrees 13' N). Punkies of the above species are small in number and do not occur all over the region. In northern and middle taiga of Evenkia (60 degrees 20'-65 degrees 45' N) punkies are very abundant and occur everywhere. 7 species C. obsoletus, C. chiopterus, C. punctatus, C. pulicaris, C. grisescens, C. fascipennis, C. helveticus are reported from this region. A relative abundance of punkies in the complex of bloodsucking Diptera during a summer period in different landscape zones is shown. Their seasonal and daily activity is shown and maximum levels of attacking man, reindeer and dog are given.     

Equine Culicoides hypersensitivity in Florida: biting midges collected in light traps near horses

Twenty-three species of Culicoides were trapped near pruritic horses during a 2-year survey in Florida. Nearly 99% of the biting midges collected were represented by Culicoides insignis Lutz, C. edeni Wirth and Blandon, C. stellifer (Coquillett), C. niger Root and Hoffman, C. haematopotus Malloch and C. venustus Hoffman. The relative contribution to the total catch by each of these species varied among collection sites. Seasonally, different species attain their largest population sizes at different times. Association of species collected in light traps with the seasonality of lesion development on the horses suggests that more than one species must be involved in causing Culicoides hypersensitivity in horses from Florida. Nulliparous females were present in most months that females of the same species were active; in some cases, species were trapped in each month of the year, particularly at the more southerly sites. The species of biting midge most likely to be involved in Culicoides hypersensitivity in horses in Florida are C. insignis, C. stellifer and C. venustus based upon their mammalophilic behaviour and seasonality.     

An investigation on the Culicoides species composition at seven sites in southern Switzerland

In the past decade, there have been regular outbreaks of bluetongue (BT) in many parts of Europe. Owing to the presence of BT disease and its vectors in countries adjacent to Switzerland, an initial entomological survey was conducted in 2003, which established the presence of several midges of the genus Culicoides (Diptera: Ceratopogonidae). Subsequently, a sentinel herd monitoring system was established with the primary entomological aim being the determination and further study of Culicoides population compositions. Insects were collected in 2005 and 2006 at seven sentinel herd sites in the south of Switzerland (canton of Ticino) near the border of Italy, using Onderstepoort-type light traps. This region is botanically and zoologically similar to the Mediterranean and is one of the warmest and most humid areas of the country, hence it is considered a potential access path for BT disease into Switzerland. Collections were made at four cattle farms, two equestrian centres and one goat farm. Sites were sampled four times per month from June to October. Traps were operated from dusk until dawn and samples were collected monthly for analysis through microscopy as well as a Culicoides imicola -specific PCR. Results confirmed the absence of C. imicola (Kieffer) and demonstrated that the potential BT virus vectors are highly abundant, notably: Culicoides obsoletus (Meigen), Culicoides scoticus (Downes & Kettle) and Culicoides dewulfi (Goetghebuer) subgenus Avaritia and Culicoides pulicaris (Linnaeus) subgenus Culicoides . These findings expand the current knowledge of Culicoides population composition in the southern part of the Switzerland. Culicoides cataneii (Clastrier), Culicoides flavipulicaris (Dzhafarov), Culicoides indistinctus (Khalaf), Culicoides nubeculosus (Meigen) and species of the Grisescens complex were reported for the first time in Switzerland.     

Bluetongue vector species of Culicoides in Switzerland

Switzerland is historically recognized by the Office Internationale des Epizooties as free from bluetongue disease (BT) because of its latitude and climate. With bluetongue virus (BTV) moving north from the Mediterranean, an entomological survey was conducted in Switzerland in 2003 to assess the potential of the BTV vectors present. A total of 39 cattle farms located in three geographical regions, the Ticino region, the Western region and the region of the Grisons, were monitored during the vector season. Farms were located in areas at high risk of vector introduction and establishment based on the following characteristics: annual average temperature > 12.5 degrees C, average annual humidity >or= 60%, cattle farm. Onderstepoort black light traps were operated at the cattle farms generally for one night in July and one night in September. A total of 56 collections of Culicoides (Diptera: Ceratopogonidae) were identified morphologically. Only one single individual of Culicoides (Avaritia) imicola, the major Old World vector of BTV, was found in July 2003 in the Ticino region, one of the southernmost regions of Switzerland. In the absence of further specimens of C. imicola from Switzerland it is suggested that this individual may be a vagrant transported by wind from regions to the south of the country where populations of this species are known to occur. Alternative potential BTV vectors of the Culicoides (Culicoides) pulicaris and Culicoides (Avaritia) obsoletus complexes were abundant in all sampled regions with individual catches exceeding 70 000 midges per trap night.     

Field studies on efficacy of host odour baits for the biting midge Culicoides impunctatus in Scotland

The efficacy of some putative attractants for the biting midge Culicoides impunctatus (Goetghebuer) (Diptera: Ceratopogonidae) was assessed using odour-baited 'delta traps' and suction traps. 1-octen-3-ol was confirmed as a potent olfactory attractant for C. impunctatus when released at 0.06mg/h. Acetone (23mg/h) and a mix of six phenolic compounds (phenol, 3-ethylphenol, 4-ethylphenol, 3-methylphenol, 4-methylphenol and 4-propylphenol), at undetermined release rate, also significantly increased delta trap catches compared to unbaited controls. When tested in combination, there was evidence of synergism between CO2 (0.2L/min) and acetone, 1-octen-3-ol or cow urine, trap catches being, respectively, 4.7, 6.2 and 9.3-fold greater than for CO2 alone. Highest catches were obtained with triple bait combinations comprising cow urine + acetone + CO2 or cow urine + 1-octen-3-ol+CO2, which increased trap catches by X 22 and X 24, respectively, compared to CO2 alone. Culicoides impunctatus was found to be extremely sensitive to CO2 and responses, gauged over two field seasons, showed a significant dose-dependent increase in catch across the entire range of release rates (0.2-2.5 L/min). Responses to these release rates, ranging from small to large mammal equivalents, emphasized the important role of CO2 in host location by C. impunctatus. Uses of olfactory attractants for monitoring and control of Culicoides are reviewed on the basis of these results.     

Presence in the Balearic Islands (Spain) of the midges Culicoides imicola and Culicoides obsoletus group

An outbreak of the livestock viral disease bluetongue (BT) was detected during September and October 2000 in the Balearic Islands, Spain. Due to the lack of information about the species of Culicoides (Diptera: Ceratopogonidae) reported in the affected area, six farms in Majorca, four in Minorca and one in Ibiza were selected to carry out surveillance of Culicoides adults using light traps. Here, for the first time, we report the presence in the Balearic Islands of Culicoides imicola Keiffer, the main vector of BT, and the Culicoides obsoletus Meigen group.