Field observations in Israel on hypersensitivity in cattle, sheep and donkeys caused by Culicoides

SUMMARY Hypersensitive lesions in the skin of sheep, cattle and donkeys from various regions in Israel are described. No cause other than insect bites was found. The animals were free of ticks, lice, fleas and blackflies, and of all the prevailing insect groups; Culicoides spp were considered as the most probable cause of the hypersensitivity. Lesions on the legs and abdomen were attributed mainly to C obsoletus Melgen, C schultzel gp and C puncticollis Becker, and less to C Imicola Kieffer. Lesions on the back, flanks, neck and head were attributed mainly to C Imicola, less to C newsteadl Austen and rarely to C pullcaris Linnaeus.     

Epidemiological study of Culicoides hypersensitivity in horses in Israel

The variations with breed, gender, age and coat colour in the prevalence of Culicoides hypersensitivity were studied in 408 horses on 18 farms in Israel. Data were gathered by means of questionnaires and the diagnoses were confirmed by direct physical examinations. The prevalence of Culicoides hypersensitivity was 28 per cent. The disease was rare on farms more than 800 m above sea level but was more prevalent at lower altitudes. The most important factors affecting the prevalence of Culicoides hypersensitivity were the farm, breed and age, but gender and colour were not significantly correlated with its prevalence.     

Culicoides species attracted to horses with and without insect hypersensitivity

The aims of this study were to determine (1) which species of Culicoides is most commonly attracted to horses, (2) whether horses suffering insect hypersensitivity attract more Culicoides spp. than unaffected horses, and (3) the times when Culicoides spp. are most active. Horses affected by insect hypersensitivity and unaffected horses were placed inside mosquito netting tents for 30 min at different times of the day. All Culicoides spp. trapped inside the tents were collected and identified. C. obsoletus was the most common species found, followed by C. pulicaris. Healthy horses attracted slightly more midges than horses that were affected with insect hypersensitivity. All of the Culicoides species were most active at sunset, less so at sunrise and very few or no midges were trapped in the afternoon or at night.     

Culicoides species associated with sheep in the Netherlands and the effect of a permethrin insecticide

Culicoides spp. act as vectors for a number of viral diseases of animals including bluetongue in sheep. The aims of this study were to determine: (1) which Culicoides spp. are associated with sheep in The Netherlands; (2) the time of the day when they are most active; and (3) the effect of treatment of animals with a permethrin insecticide. Two pairs of sheep were each housed within mosquito tents of either one or two layers of netting and all trapped Culicoides spp. were identified microscopically. For the permethrin insecticide study, one of two pairs of sheep was treated with 3.6% permethrin and all animals were housed in tents of similar design. Of the 6210 midges captured, 54.1% were identified as C. chiopterus and 42.7% as C. obsoletus. C. imicola was not identified. The average insect feeding rate was 35-40% and midge activity was greatest around sunset. Permethrin treatment reduced the number of midges captured by 50% and also resulted in a decrease in the percentage of midges that had fed. The findings provide useful information on the behaviour and distribution of Culicoides spp. that will facilitate the development of appropriate control strategies to minimise the risk of insect-vector borne virus diseases such as bluetongue.     

A histamine release assay to identify sensitization to Culicoides allergens in horses with skin hypersensitivity

Skin hypersensitivity is an allergic disease induced in horses by allergens of Culicoides midges. The condition is typically diagnosed by clinical signs and in some horses in combination with allergy testing such as intradermal skin testing or serological allergen-specific IgE determination. Here, we describe an alternative method for allergy testing: a histamine release assay (HRA) that combines the functional aspects of skin testing with the convenience of submitting a blood sample. The assay is based on the principle that crosslinking of allergen-specific IgE bound via high-affinity IgE receptors to the surfaces of mast cells and basophils induces the release of inflammatory mediators. One of these mediators is histamine. The histamine was then detected by a colorimetric enzyme-linked immunosorbent assay. The histamine assay was used to test 33 horses with skin hypersensitivity and 20 clinically healthy control animals for histamine release from their peripheral blood basophils after stimulation with Culicoides allergen extract or monoclonal anti-IgE antibody. An increased histamine release was observed in the horses with skin hypersensitivity compared to the control group after allergen-specific stimulation with Culicoides extract (p=0.023). In contrast, stimulation with anti-IgE induced similar amounts of released histamine in both groups (p=0.46). For further evaluation of the HRA, we prepared a receiver operating-characteristic (ROC) curve and performed a likelihood-ratio analysis for assay interpretation. Our results suggested that the assay is a valuable diagnostic tool to identify sensitization to Culicoides allergens in horses. Because some of the clinically healthy horses also showed sensitization to Culicoides extract, the assay cannot be used to distinguish allergic from non-allergic animals. The observation that sensitization is sometimes detectable in non-affected animals suggested that clinically healthy horses use immune mechanisms to control the reaction to Culicoides allergens that are different or absent in allergic horses.     

Immunopathogenesis and immunotherapy of Culicoides hypersensitivity in horses: an update

Culicoides hypersensitivity (CH) is a strongly pruritic dermatitis of horses, characterised histologically by mixed perivascular to diffuse cellular infiltrates of mononuclear cells and eosinophils, and caused by helper T cell (Th)2, immunoglobulin (Ig)E-mediated reactions to antigens present in the saliva of biting Culicoides midges. As treatment of CH is still not satisfactory, a better knowledge of the pathogenesis of CH is needed for the development of new therapeutic modalities. Accordingly, availability of pure Culicoides allergens instead of crude whole body extracts is essential for the improvement of in vitro diagnosis of CH and development of a more effective allergen immunotherapy (AIT). Beside the Th2 cytokines interleukin (IL)-4 and IL-13, recent studies have shown the involvement of thymic stromal lymphopoietin, IL-5 and IL-31 in the pathogenesis of CH. The importance of IL-5 and IL-31 has been confirmed through targeting these cytokines by active immunisation of CH patients, resulting in a significant decrease of CH lesion scores. Thirty Culicoides salivary allergens have been described over the last 10 years and produced as pure recombinant (r-) proteins. Use of a protein microarray has allowed the identification of the most relevant Culicoides r-allergens for CH in central and northern Europe. An immunisation protocol has been developed for preventive AIT against CH. Small amounts of r-Culicoides allergens injected into the submandibular lymph nodes with Alum/MPLA as adjuvants seem to induce a suitable immune response for both preventive and therapeutic AIT. The next years will reveal whether these recent advancements will translate into an improved diagnosis, treatment and prevention of CH.     

Zinc and copper plasma levels in Icelandic horses with Culicoides hypersensitivity

Zinc concentration has been shown to have a potent immunomodulatory capacity, particularly influencing T helper cell organisation and cytokine secretion. Culicoides hypersensitivity (CHS) in horses resembles the early and late phase of type I hypersensitive reactions in man, characterised by a shift from T helper cell subtype 1 to T helper cell subtype 2 cytokine profile. In this pilot study, zinc and copper levels were measured in the plasma of 48 CHS-affected and 56 healthy Icelandic horses age 4-25 years (mean approximately 11 years) kept on 7 farms. Affected horses were divided into 3 groups according to the severity of disease. Time of blood collection and feeding management was constant. No differences in zinc or copper plasma levels and plasma copper/zinc ratio were determined among CHS horses and controls by univariate analysis of variance. Therefore, the most significant influences on zinc and copper plasma levels were affected by the location of housing. However, Spearman correlation showed a negative coefficient between the plasma zinc concentration and the severity of CHS (r = -0.31). Due to a probability value of P = 0.002 the null hypothesis r = 0 is rejected, although only 9% of the total variation of plasma zinc is presently explained by its relationship to CHS. In contrast, the Spearman correlation coefficient between plasma copper levels and severity of CHS was not significant (r = -0.14; P = 0.16). The minor deviations in plasma zinc concentrations in association with the severity of CHS may be real or due to neurohumoral or cytokine-mediated mechanisms, but appear too minimal to be relevant.     

Culicoides hypersensitivity in the horse: 15 cases in southwestern british columbia

The investigation of a chronic, seasonal dermatitis of horses in southwestern British Columbia is described. Typically the history indicated an insidious onset, followed by a gradual progression in the severity of the signs each year. Lesions appeared during the warmer months of the year and tended to regress during the winter. The clinical signs consisted of areas of pruritus and excoriation, affecting predominantly the ventral midline, mane and tailhead. In all cases corticosteroid therapy relieved the pruritus and allowed the lesions to heal.

The salient pathological findings were hyperkeratosis, spongiosis and a dermal infiltration of eosinophils together with mononuclear cells. These changes are typical of an allergic dermatitis, which has been recognized in many parts of the world as a hypersensitivity reaction to the bites of Culicoides spp. In this instance, the epidemiological findings relating to the geographic area, the local insect population and the distribution of lesions implicated Culicoides obsoletus as the etiological agent.

     

Seasonal allergic dermatitis in sheep associated with Ctenocephalides and Culicoides bites

The clinical, epidemiological and histopathological findings of two pruritic dermatites in sheep in Israel are described. The first type of dermatitis affected mainly young animals with lesions predominantly on the legs. It occurred from March to November, with a peak in June. The second type affected animals of all ages and was mainly on the ventrum. It was sporadic but occurred throughout the year with a peak in October. The morbidity rate of this syndrome reached 4.3% in one flock. The histopathology of both conditions was consistent with an allergic dermatitis. Fleas and midges were collected and identified as Ctenocephalides felis felis and various species of Culicoides. The population density, seasonal activity, geographical distribution and feeding behaviour preferences of the insects and the incidence of the two types of dermatitis suggest that fleas and midges were the causal agents. Flea and midge bite pruritic dermatoses should be considered in the differential diagnosis of sarcoptic and psoroptic mange.     

Serial cyclic transmission of bluetongue virus in sheep and Culicoides variipennis.

Bluetongue virus (strain 62-45S) was transmitted from sheep to sheep throughout a year by vector bites. A colonized population (SONORA strain, 000 line) of the biological vector Culicoides variipennis (Coquilllett) was used. Fifteen serial cyclic transmissions covered a period of 13 months from October through November of the following year. The mean infection rate of the biting gnats was 37 percent. The clinical response to bluetongue virus was significantly more severe in sheep infected by vector bites than in those inoculated with the virus at the same sheep-serial passage level. A second corroborative serial transmission was conducted for 7 months from June through December. The mean infection rate of the vector was 27%.     

Equine Culicoides hypersensitivity: evaluation of a skin test and of humoral response

Intradermal tests were carried out on 18 horses with clinical signs of Culicoides hypersensitivity (CHS) and 23 horses without clinical signs of CHS, and sera from these horses were analysed by SDS-PAGE and Western blotting (W-B). Intradermal injections of 0.1 ml of 25 microg/microl sterile Culicoides extract, 0.1 ml of 1:10,000 histamine (positive control) and 0.1 ml of physiological saline (negative control) were made in the dermis of the middle region of the neck. Analysis of reactions indicated that a 1 cm wheal and a skinfold thickness >10% at 24 h represented a valid cut-off between horses with and without CHS. In these conditions the test, even in winter when clinical signs were absent, had 100% sensitivity and specificity. The W-B was performed after running Culicoides extract on a 12% polyacrylamide gel. The test revealed the presence of several bands with molecular weight ranging from 6 to 200 kDa. In particular, a band of 65 kDa was predominantly found in hypersensitive horses by using an anti-IgE antibody while in normal horses the same band was mainly detected by using an anti-IgG antibody. Our results demonstrated that the skin test is a valid diagnostic test, with high sensitivity and specificity and that the band of about 65 kDa probably corresponds to the allergen involved in the pathogenesis of CHS.     

Comparison of black and white light for collecting Culicoides imicola and other livestock-associated Culicoides species in South Africa

Comparison of the effectiveness of 8W fluorescent black and white light sources, in two 4x4 Latin squares (16 replicates) designs under South African conditions, showed black light to be up to three time more effective in collecting Culicoides imicola Kieffer (Diptera, Ceratopogonidae) and other South African Culicoides species. Four Culicoides species, which were collected in low numbers with black light, were not collected in traps equipped with the white light source. No significant difference was found in the parous rate of the C. imicola populations as determined by the two light sources. The study highlighted the superiority of black light as a preferred collection method for C. imicola, considered to be the most widespread and abundant vector of livestock orbiviruses. The results underline the need to develop and adopt standard techniques for measuring the variables of vectorial capacity.