Preferential landing and engorging sites of Culicoides species landing on a horse in Ireland

An investigation of the landing and engorging sites of midges on a bait horse was made in Ireland as part of a project examining the aetiology of sweet itch. Ten species were recovered, seven of which engorged. Culicoides obsoletus and C dewulfi were the most commonly represented, accounting for 89.9 per cent of midges landing and 90.5 per cent of midges engorging. Only 4.69 per cent of midges attracted to the bait were C pulicaris. Overall, the preferential landing sites for midges were along the mane and lower leg regions. Few culicoides were attracted to the head, front and underside of the animal. Two species, C punctatus and C nubeculosus, fed exclusively at the predilection sites of sweet itch but in very small numbers. C pulicaris fed predominantly at these sites also and represented 5.08 per cent of the engorging catch. Of 10,178 midges attracted to the bait, 4097 (40.3 per cent) engorged; the majority of the species taking full blood meals fed on the mane and lower legs. Midges biting in other areas tended to have smaller meals. Areas along the dorsal midline of the body are predilection sites for sweet itch lesions and are preferential engorging sites also. However, considerable engorgement occurred on the lower legs where sweet itch lesions do not occur.     

Potential of infra-red thermography for the detection of summer seasonal recurrent dermatitis (sweet itch) in horses

The feasibility of using infra-red thermography for early and out of season detection of summer seasonal recurrent dermatitis (sweet itch) was tested on 13 sensitive and six insensitive horses in Israel. In summer (June, July and September) six affected mares and two unaffected sensitive mares (one of them a pony) were clearly 'warmer' in the affected zones than three insensitive mares. In winter (February and March) unaffected sensitive horses could be differentiated from insensitive mares by the warm areas detected in the affected zones. Small numbers of Culicoides imicola bites may have been the cause of the local heat reactions of the body that were detected by the infra-red radiometer.     

Seasonal allergic dermatitis in sheep in southern Brazil caused by Culicoides insignis (Diptera: Ceratopogonidae)

The incidence of seasonal dermatitis was studied in a Hampshire Down flock on a farm in southern Brazil. Epidemiological data, clinical signs and macroscopic pathology were obtained by visiting the farm. Histological lesions were studied in skin biopsies of affected sheep. Biting insects were collected from January to April 2005 in an attempt to identify the etiological agent of the disease. Disease prevalence was 40%; the age of the affected animals was variable. Disease occurred from December to March, some animals had lesions for the entire year. Clinical signs include pruritus on the ears, around the eyes and ventral abdomen. Initially erythema and small red papules were seen, followed by alopecia and crust formation. Histologically the lesions were characterized by perivascular eosinophilic dermatitis. Hyperkeratosis and acanthosis were observed in the chronic lesions. Both Anopheles albitarsis and Culicoides insignis were captured during the study. C. insignis bites caused pruritus in sheep. Both types of insects were caught when they approached the sheep baits approximately 30min after sunset. Results suggested that the disease occurred as a result of an immediate hypersensitivity reaction to C. insignis.     

Immunogenicity and allergenicity of Culicoides imicola (Diptera: Ceratopogonidae) extracts

Summer seasonal recurrent dermatitis (SSRD) or "sweet itch" is a seasonally occurring allergic dermatitis of horses provoked by biting midges. The allergic skin reactions have been attributed to allergens present in various Culicoides species. C. imicola is the suspected etiological agent of SSRD in Israel. Whole body extracts of this midge induced hypersensitivity reactions upon injection into susceptible horses and in this study attempts were made to define components of C. imicola which have immunogenic and allergenic properties. Immunogenic potency was evaluated by raising antisera to whole body extracts of C. imicola in rabbits and examining their reactivity towards fractionated extracts. Allergenic potency was examined by reacting fractionated extracts with horse sera. Humoral reactivity of susceptible and non susceptible horses was assayed by specific IgE and IgG ELISAs. Although there are many antigenic components in whole body extracts of C.imicola capable of eliciting an immune response, no conclusive evidence was obtained indicating that allergic reactivity was associated with increased IgE levels of defined specificity.     

Papular dermatitis induced in guinea pigs by the biting midge Culicoides sonorensis (Diptera: Ceratopogonidae).

Histological, ultrastructural, and virological examinations were performed on abdominal skin from guinea pigs after a blood meal by colony-bred biting midges, Culicoides sonorensis. Small, superficial, cutaneous, crateriform ulcers with necrosis of superficial dermis developed at feeding sites and healed within 24-48 hours. Animals developed nonpruritic erythematous papules 5 days after feeding that persisted until the study ended at 12 days after feeding. Papules corresponded histologically to foci of epidermal hyperplasia and superficial interstitial dermatitis with intraepidermal micropustules and scattered intraepidermal polykaryons. The principal ultrastructural changes were spongiosis in germinal epithelium and neutrophilic-histiocytic exocytosis. No viral agents or broken mouthparts were identified in lesions. The dermatitis may represent a host reaction to persisting insect salivary secretion and should be considered as an additional consequence of blood feeding in future studies involving biting midges.     

Chemical composition of silage residues sustaining the larval development of the Culicoides obsoletus/Culicoides scoticus species (Diptera: Ceratopogonidae)

Culicoides (Diptera: Ceratopogonidae) are biological vectors of bluetongue virus (BTV). Bluetongue is a viral disease that affects domestic and wild ruminants. Since its recent emergence in northern Europe, this disease has caused considerable economic losses to the sheep and cattle industry. The biotopes, and more particularly the chemical characteristics which are suitable for larval development of the main vector species, are still relatively unknown. This study shows that the larvae of biting midges belonging to the species Culicoides obsoletus and Culicoides scoticus are able to breed in different types of silage residue (maize, grass, sugar beet pulp and their combinations). The chemical composition of substrates strongly influences the presence of the immature stages of these biting midges. Higher lignin and insoluble fibre contents seem to favour their presence and could play the role of a physical support for semi-aquatic larvae. In contrast, higher concentrations of magnesium and calcium are negatively correlated with the presence of these two species. These data will help to locate and monitor the breeding sites of these species and could contribute to the control of these insects on farms.     

Abundance of biting midge species (Diptera: Ceratopogonidae,Culicoides spp.) on cattle farms in Korea

Culicoides biting midges were collected on three cattle farms weekly using light traps overnight from May to October between 2010 and 2011 in the southern part of Korea. The seasonal and geographical abundance of Culicodes spp. were measured. A total of 16,538 biting midges were collected from 2010 to 2011, including seven species of Culicoides, four of which represented 98.42% of the collected specimens. These four species were Culicodes (C.) punctatus (n = 14,413), C. arakawae (n = 1,120), C. oxystoma (n = 427), and C. maculatus (n = 318). C. punctatus was the predominant species (87.15%).     

Culicoides arakawae (Diptera: Ceratopogonidae) population succession in relation to leucocytozoonosis prevalence on a chicken farm in Taiwan

Leucocytozoonosis caused by Leucocytozoon caulleryi is a significant disease prevalent in open chicken houses of southern and eastern Asia. L. caulleryi is transmitted by Culicoides arakawae, a blood-sucking vector. Leucocytozoonosis prevalence is influenced by vector population succession. Thus this examination was performed on a farm to investigate vector population succession and leucocytozoonosis prevalence in experimental chicks and to obtain the ecology data for assessing the prevalence. The findings were as follows: (1) C. arakawae adults might be highly host specific because they were rarely discovered on cattle or pig farms, and none of the experimental chickens were infected by L. caulleryi on those farms. (2) Identifying and counting gorged and gravid C. arakawae to assess leucocytozoonosis prevalence is a practical strategy. The critical vector index should be 5.0 calculated by dividing the smallest vector mean from the prevalent period by the largest vector mean from the population not causing leucotocytozoonosis. (3) Taking vector means from three or more collections each month, should be the best assessment of leucocytozoonosis prevalence because C. arakawae succession appears to have a 3-week periodicity. Hopefully, these findings will contribute to assessing leucocytozoonosis prevalence.     

Evaluation of Culicoides insignis (Diptera: Ceratopogonidae) as a vector of bluetongue virus.

Based upon epidemiological evidence, Culicoides insignis Lutz is a probable biological vector of bluetongue viruses (BTV) in South Florida, the Caribbean Region and Central America. The vector potential of this species for BTV was evaluated in the laboratory in a series of experiments using insects caught in the field. Although there was great variation in the percentage of flies that fed from any one catch, it was demonstrated that C. insignis became infected after membrane feeding on a mixture of blood and virus. The infection rates ranged from 20 to 62.5%. Following intrathoracic inoculation, BTV replicated to high titres in C. insignis. Such flies were also shown to be capable of transmitting BTV to susceptible sheep and embryonated chicken eggs. This series of experiments provides the first conclusive evidence that C. insignis is a biological vector of bluetongue virus. This is the first proven vector of BTV in the neotropics.     

An early record of Culicoides species (Diptera: Ceratopogonidae) developing in the dung of game animals in southern Africa

The value of the dung of game animals as a larval habitat for Culicoides species was briefly investigated in 1973/74 in the Kruger National Park, Transvaal, Republic of South Africa. A total of 4 species in the subgenus Avaritia was reared, 3 from elephant dung, 2 from zebra dung and 1 from the dung of buffalo. These early studies have recently led to more intensive rearing and a better understanding of the systematics of this subgenus in the Afrotropical region.     

Culicoides spp. (Diptera: Ceratopogonidae) Associated with Farmed Ostriches (Struthio camelus) in Botswana

Veterinary Research Communications -     

Culicoides arakawae (Diptera: Ceratopogonidae) efficiently blood-fed and infected with Leucocytozoon caulleryi through a natural membrane

Culicoides arakawae, the most common Culicoides sp. on chicken farms in East Asia, is an important blood-sucking insect and Leucocytozoon caulleryi vector. How parasites, in an ingested blood bolus, enter the midgut of insects and deal with this complex and biochemically hostile environment is poorly understood. However, successful blood-feeding through a membrane in C. arakawae is beneficial for studying this phenomenon. Therefore, a membrane-feeding method for C. arakawae was developed in. The blood-feeding success rates of C. arakawae fed through five different membranes were: turkey egg at 43.7+/-11.7%, chicken egg at 45.2+/-12.1%, duck egg at 38.8+/-12.0%, pig gut at 0% and chick skin at 0%. In fertility measurements, the average number of eggs produced for C. arakawae fed through egg-shell membrane, at 77.7+/-15.1 per female, was significantly higher (P<0.01) than the 46.7+/-10.6 found in C. arakawae fed on the breast skin of a live chicken. Meanwhile, in parasite infectivity tests, C. arakawae could be infected by L. caulleryi when the vector was blood-fed with infective blood cells reconstituted with specific pathogen-free (SPF) sera through an egg-shell membrane. The sporozoite average and infection rates of inoculated chicks were 166.8+/-12.5 and 100%, respectively. In conclusion, feeding C. arakawae blood through fowl-egg-shell membranes should be an efficient method for in vitro infection of midges as the engorged midges are infected by parasites and display reproductive potential. Furthermore, the method is practical for feeding a large number of midges.     

Bluetongue virus isolations from midges belonging to the Obsoletus complex (Culicoides, Diptera: Ceratopogonidae) in Italy

Between July and September 2002 there were outbreaks of bluetongue on three sheep holdings in the communities of San Gregorio Magno (Salerno, Campania), Laviano (Salerno, Campania) and Carpino (Foggia, Puglia), and the involvement of bluetongue virus (btv) was confirmed serologically and virologically. The mortality rate was at least 11 per cent and involved btv serotype 2 (btv-2) and serotype 9 (btv-9). These holdings were also surveyed for the Culicoides (Diptera: Ceratopogonidae) vectors; approximately 10,000 midges belonging to 15 species were captured, but they did not include a single specimen of the classical Afro-Asiatic bluetongue vector, Culicoides imicola. Species belonging to the Obsoletus complex dominated the light-trap collections, and Culicoides obsoletus Meigen, Culicoides scoticus Downes and Kettle and Culicoides dewulfi Goetghebuer constituted 90 per cent of all the Culicoides species captured. Fifty-six pools of the Obsoletus complex (excluding C dewulfi), each containing 100 individual midges and containing only parous and gravid females, were assayed for virus. btv-2 was isolated from three pools from San Gregorio Magno and Carpino, and btv-9 was isolated from one pool from Laviano. These results indicate that a species other than C imicola is involved in the current re-emergence of bluetongue in the Mediterranean Basin, but whether it is C obsoletus sensu stricto or C scoticus, or both, is uncertain.     

Rearing of Culicoides nubeculosus (Diptera: Ceratopogonidae) by natural or artificial feeding in the laboratory

A technique for the mass rearing of Culicoides nubeculosus in the laboratory is described. Female midges were fed either on fresh or deep-frozen, defibrinated cattle blood (-70 degrees C) through latex membranes, or on anesthetized white mice. Feeding rates of up to 90% were obtained on the latex membrane, whereas only 41% of the midges fed on mice. The best oviposition rates of greater than 50% were achieved after feeding either on the latex membrane with fresh cattle blood or on mice. An average of greater than 100 eggs per female were deposited. The highest larval hatching rate was observed after feeding with fresh blood; about half of the larvae developed to the adult stage. A reproduction index was defined for the colony based on the feeding rate, oviposition rate, larval hatching rate and development to the adult stage. The highest reproduction index was obtained when the midges were fed on fresh cattle blood through the membrane.     

Simultaneous quantification of the relative abundance of species complex members: application to Culicoides obsoletus and Culicoides scoticus (Diptera: Ceratopogonidae), potential vectors of bluetongue virus

The two sympatric sibling species Culicoides obsoletus (Meigen) and Culicoides scoticus Downes and Kettle (Diptera: Ceratopogonidae), are known to be competent vectors for bluetongue virus in the Palaearctic region. However, morphological identification of constituent species is only readily applicable to adult males and these two species distinguishing traits have overlapping character states. As their vector competence may differ in space and time, the correct identification and quantification of specimens of both species are essential for understanding bluetongue epidemiology. However, no molecular tools are available for high-throughput identification of the two species. We therefore developed a quantitative duplex real-time PCR assay to determine the relative abundance of each sibling species in a sample using TaqMan probes. For each species, standard curves were constructed from serial dilutions of purified plasmid DNA containing ITS1-5.8S-ITS2 (rDNA) in the range of 10(-1) to 10(-5)ng/μL. Standard curves were used to quantify samples of mixed C. obsoletus/C. scoticus specimens. Specificity was evaluated with 5156 specimens representing 62 species. Based on the DNA quantities detected according to the standard curves, a quadratic model developed on 1100 males and validated on 555 females was able to predict the relative abundance of each species simultaneously in a one-shot reaction (Pearson coefficient of 0.999). Our assay showed a requirement of two specimens or less for 95% of the predictions, making it highly applicable to field collections. Extensive use of this real-time PCR assay will provide a better understanding of geographical distribution, dynamics, and bionomics on a species level, which is essential for risk assessment. This approach is an important contribution to medical entomology for investigating the vector role of arthropod sibling species.     

Allergic dermatitis (sweet itch) of Icelandic horses in Sweden: an epidemiological study

A survey of allergic dermatitis (sweet itch) in Sweden contained information on 441 Icelandic horses. Results of a questionnaire indicated that approximately 15 per cent of the country's Icelandic horses suffered from the disease. The prevalence of allergic dermatitis was significantly higher among horses imported from Iceland (26.2 per cent) compared to that of Swedish-born animals (6.7 per cent). In addition, horses born in Iceland were significantly more severely affected than horses born in Sweden. The risk of allergic dermatitis in Sweden appeared to be more than six times higher for horses exported from Iceland to Sweden relative to that of horses originally born in Sweden. The prevalence of disease for horses of seven years or older was 30 per cent for Icelandic-born individuals as compared to 7.3 per cent for horses born in Sweden. Similarly, the risk of allergic dermatitis in Sweden for horses of seven years or older appeared to be nearly 10 times higher for horses imported from Iceland relative to that of horses born in Sweden. Allergic dermatitis usually appeared during the third grazing season for imported horses and during the fourth season for horses born in Sweden. Furthermore, the course of the disease tended to become worse with time. Analysis of the prevalence of allergic dermatitis relative to gender revealed no significant differences. Certain geographical variations in the prevalence of the disease was also found.