Establishment of a mouse allergy model for Culicoides (Diptera: Ceratopogonidae).

BACKGROUND: Culicoides is a genus of ubiquitous biting midges (Ceratopogonidae). Female midges have blood-sucking habit. They not only bite and harass humans and animals but also may be an important vector of disease transmission. Therefore, building an animal allergy model caused by Culicoides biting is very beneficial for studying its pathogenesis and exploring the therapeutic methods. MATERIAL AND METHOD: Kunming mice were used in this study to build the model and sensitised by two-step injection of midge extracts. Scratching behaviour and histological examination were used to check the immediate and delayed responses. Immunoglobulin E (IgE) and Immunoglobulin G (IgG) were detected using indirect enzyme-linked immunosorbent assay (ELISA) assay. Splenic cell proliferation and cytokine production were determined using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and ELISA assays. The response of cytokine gene expression to midge stimulation was analysed through quantitative real-time polymerase chain reaction (qPCR). RESULTS: Behavioural results revealed a significant increase in scratching frequency among the midge-sensitised animals (p < 0.05). Histological examination showed more inflammatory cytokine infiltration at the injection site of midge-sensitised mice comparing to the ones in the control group. The serum levels of IgE and IgG1 antibodies in the midge-sensitised group were significantly elevated (p < 0.05). After splenocytes were stimulated in vitro with midge extracts, the midge-sensitised group's splenocyte count significantly increased in comparison to the control group. The midge-sensitised group's qPCR data revealed a down-regulation of tumor necrosis factor alpha (TNF-α) expression and an increase in the expression of interleukin (IL)-4, IL-5, IL-10 and IL-13 but not in the control group (p < 0.05). CONCLUSIONS: In this study, an animal model of Culicoides-mouse sensitisation was successfully constructed using a two-step method. The mode of administration of the model was in good agreement with the natural immune pathway, and the immune response induced by the sensitisation of the model was similar to that produced by the bite of a midge.

Oral Calcinosis Circumscripta Associated with Wasp (Hymenoptera vespidae) Sting in a Cat.

In veterinary medicine, calcinosis circumscripta is a morphological diagnosis for which the cause may be known or idiopathic. The aetiology of the lesion must be investigated because the nature of the underlying disease process will dictate appropriate treatment. We present a case of oral calcinosis circumscripta in a cat caused by a wasp sting and associated with moderate unilateral facial oedema. Following corticosteroid therapy and oedema resolution, hyperaemic, circular, firm, elevated lesions were observed on the buccal mucosa and evolved into whitish, slightly elevated multifocal plaques with an irregular surface. As antibiotic and corticosteroid therapy was ineffective, an excisional biopsy was performed after 6 months of lesion development and a diagnosis of calcinosis circumscripta of dystrophic aetiology was made. To our knowledge, this is the first report of calcinosis circumscripta in association with an insect sting in an animal. Insect stings should be included in the differential diagnosis of calcinosis circumscripta in cats.

Anaphylaxis-related hemoperitoneum in 11 dogs.

OBJECTIVE: To describe the unique complication of hemoperitoneum associated with anaphylaxis. DESIGN: Retrospective case series from September 2012 to August 2017. SETTING: Two private emergency and specialty referral hospitals. ANIMALS: Eleven client-owned dogs diagnosed with anaphylaxis and hemoperitoneum upon presentation or referral. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Inclusion criteria included clinical signs consistent with anaphylaxis (hypotension, tachycardia, vomiting, diarrhea, weakness, collapse, with or without the presence of dermal signs) due to witnessed or unwitnessed presumed bee sting, an elevated alanine aminotransferase (ALT), performance of abdominal FAST (AFAST) examination with an abdominal fluid score, the sonographic presence of gallbladder wall edema, and hemoperitoneum. All dogs (n=11) were managed medically without surgical intervention. 91% (n=10) of dogs survived to discharge. CONCLUSIONS: Hemoperitoneum development can be seen with anaphylactic reactions, though the exact mechanism remains to be fully understood. Medical therapy is warranted and can be successful in these patients; surgery is not indicated to address hemoperitoneum.

Aedes (Stegomyia) aegypti mosquito bite hypersensitivity in a dog: a case report.

BACKGROUND: Mosquitoes are vectors of several pathogens of considerable importance to humans and companion animals, including nematode helminths such as Dirofilaria immitis and Dirofilaria repens that cause heartworm disease and subcutaneous dirofilariosis, respectively. In addition to mosquito-borne pathogen transmission, mosquito bites can cause discomfort and irritation in pets, and even lead to severe hypersensitivity reactions. In the present study, we report an acute local hypersensitivity reaction in a dog following experimental exposure to Aedes (Stegomyia) aegypti. CASE PRESENTATION: A healthy six-year-old male beagle was included in an efficacy study in which dogs (n = 28) were exposed to Ae. aegypti mosquitoes. On Day - 6, the dog was allocated to one of the study groups, consisting of seven dogs to be treated on Day 0 with an imidacloprid/flumethrin collar. After sedation, animals were exposed to approximately 50 females of Ae. aegypti for 60 (± 5) minutes on Days - 6, 1, 7, 14, 21, 28, 55, and 83. On Day - 6, no allergic reaction to the mosquito bites was observed. However, on Day 1, corresponding to the second challenge, the dog demonstrated an acute allergic reaction characterized by swelling of the face (especially in the base of the muzzle and around the eyes), redness of the eyes, and conjunctival edema of the right eye was also observed. The dog was immediately treated with an intramuscular injection of a commercially available antihistamine treatment, Pen-Hista-Strep® containing a suspension of benzylpenicillin, chlorphenamine, dexamethasone, dihydrostreptomycin, and procaine at a dosage of 1 mL per 10 kg. A few hours after treatment, the dog showed noticeable improvement. CONCLUSIONS: This case provides the first evidence of canine acute local hypersensitivity reaction to mosquito bites under laboratory conditions. This observation suggests that invasive mosquito species such as Aedes spp. may affect the health and comfort of our companion animals, especially for pets with outdoor access without individual protective measures against insect bites.

Essential oil spray reduces clinical signs of insect bite hypersensitivity in horses.

OBJECTIVE: To assess the efficacy of an herbal spray combining various essential oils, with a claim of mast cell stabilisation, antipruritic, anti-inflammatory, and insect repellent effects on the clinical presentation of insect bite hypersensitivity (IBH) in horses. DESIGN: Double-blinded, placebo-controlled, randomised, cross-over clinical trial. METHODS: Twenty adult horses with clinical IBH were treated with a daily application of herbal spray or placebo for 28 days in a randomised, cross-over fashion, separated by a>28-day washout period. Horses were examined and scored prior to and after the completion of each treatment. Histopathology was performed on four horses. Owners kept daily diaries of observations. RESULTS: The herbal spray significantly reduced the severity of all assessed parameters (pruritus, excoriations, lichenification and alopecia; P < 0.05) compared with baseline values (pretreatment) and with placebo. Owners reported improvement of pruritus in 19/20 horses (95%) with complete resolution in 17 horses (85%) following treatment. Skin biopsies showed resolution of orthokeratosis in 4/4 horses, reduced thickness of the stratum spinosum in 2/4 horses and complete resolution of histopathological abnormalities in 1/4 horses after treatment, compared with either no change or deterioration of histopathologic lesions after placebo. No side effects were observed. CONCLUSIONS: The tested herbal spray may be an effective treatment for the management of equine IBH.

Better than DEET Repellent Compounds Derived from Coconut Oil.

Hematophagous arthropods are capable of transmitting human and animal pathogens worldwide. Vector-borne diseases account for 17% of all infectious diseases resulting in 700,000 human deaths annually. Repellents are a primary tool for reducing the impact of biting arthropods on humans and animals. N,N-Diethyl-meta-toluamide (DEET), the most effective and long-lasting repellent currently available commercially, has long been considered the gold standard in insect repellents, but with reported human health issues, particularly for infants and pregnant women. In the present study, we report fatty acids derived from coconut oil which are novel, inexpensive and highly efficacious repellant compounds. These coconut fatty acids are active against a broad array of blood-sucking arthropods including biting flies, ticks, bed bugs and mosquitoes. The medium-chain length fatty acids from C8:0 to C12:0 were found to exhibit the predominant repellent activity. In laboratory bioassays, these fatty acids repelled biting flies and bed bugs for two weeks after application, and ticks for one week. Repellency was stronger and with longer residual activity than that of DEET. In addition, repellency was also found against mosquitoes. An aqueous starch-based formulation containing natural coconut fatty acids was also prepared and shown to protect pastured cattle from biting flies up to 96-hours in the hot summer, which, to our knowledge, is the longest protection provided by a natural repellent product studied to date.

Skin lesions in Aubrac cows strongly associated with fly bites (Haematobia irritans).

BACKGROUND: The horn fly Haematobia irritans is known to cause problems in cattle including weight loss and decreased milk production; cutaneous lesions have not been widely reported and descriptions of the clinical manifestations are quite variable. HYPOTHESIS/OBJECTIVES: This study describes the clinical and histopathological lesions of several Aubrac cows that presented with focal to confluent areas of alopecia, skin scaling and thickening, suspected to be induced by H. irritans bites. ANIMALS: Twenty seven, three- to six-year-old Aubrac cows, kept in mountain pastures in France, that presented with an apparently asymptomatic alopecic dermatosis. METHODS: Samples for skin cytological evaluation, skin scrapings and trichograms were examined; also, skin biopsy specimens from affected animals were used for histopathological examination. RESULTS: Clinical lesions consisted of focal to coalescing, symmetrical areas of hair loss and scaling, located on the back, flanks, and lateral and caudal aspects of the thighs. Histopathological examination revealed perivascular, oedematous, eosinophilic dermatitis with eosinophilic folliculitis and furunculosis. Species identification of several flies captured on the cows revealed H. irritans. The final clinicopathological diagnosis was eosinophilic dermatitis and folliculitis and furunculosis, suspected to be due to H. irritans fly-bite. CONCLUSION AND CLINICAL IMPORTANCE: Fly-bite (H. irritans) dermatitis should be included in the differential diagnosis of alopecic dermatoses in cows.

Dermatite alérgica à picada de Culicoides em muar: relato de caso / Allergic dermatitis due to bite of Culicoides in muar: case report

A dermatite alérgica à picada de ectoparasitos é uma enfermidade alergoparasitária bastante comum entre animais domésticos, sendo relatada principalmente em pequenos ruminantes e em animais de companhia. Contudo, a doença é pouco diagnosticada na clínica de equídeos devido a similaridades nosológicas com outras dermatopatias. Objetivou-se, com este relato de caso, descrever a síndrome clínica, o plano diagnóstico e a conduta terapêutica de um muar acometido por essa enfermidade. Atendeu-se, no Hospital Veterinário da Universidade Federal Rural do Pernambuco, uma mula de oito anos de idade, que apresentava lesões cutâneas pápulo-crostosas e pruriginosas com evolução clínica de dois anos. Em três situações anteriores, a doença havia sido tratada como dermatite fúngica por outros médicos veterinários. Para o diagnóstico, foram solicitados exame citopatológico e parasitológico de pele, cultivo bacteriológico e fúngico, análise histopatológica e hemograma. Os exames demonstraram uma dermatite superficial perivascular eosinofílica crônica, sendo indicada a terapia tópica com dimetilsufóxido, sulfadiazina, ureia e vitamina A. O protocolo terapêutico mostrou-se satisfatório, permitindo completa remissão do quadro clínico. Este trabalho relatou achados clínicos e patológicos da dermatite alérgica à picada de Culicoides spp. em muar, além de alertar sobre a importância de exames complementares para a realização do diagnóstico diferencial e para o direcionamento terapêutico adequado.(AU)

Allergic dermatitis to ectoparasite bites is a common parasitic disease among domestic animals, being reported mainly in small ruminants and companion animals. However, the disease is poorly diagnosed in equine clinics due to nosological similarities with other skin diseases. The aim of this case report was to describe the clinical syndrome, the diagnostic plan and the therapeutic management of a mule affected by this disease. An 8-year-old mule was observed at Universidade Federal Rural de Pernambuco, presenting papular-crusted and pruritic cutaneous lesions with clinical evolution of two years. In three previous situations, the disease had been treated as fungal dermatitis by other veterinarians. For the diagnosis, cytopathological and parasitological examination of the skin, bacteriological and fungal culture, histopathological analysis and blood count were performed. The exams showed a chronic eosinophilic perivascular superficial dermatitis. A topical therapy with dimethyl sulfoxide, sulfadiazine, urea, and vitamin A was indicated. The therapeutic protocol was satisfactory, allowing complete remission of the clinical condition. This work reported clinical and pathological findings of allergic dermatitis to the bites of Culicoides spp. in muar, in addition to alerting about the importance of complementary examinations for the accomplishment of the differential diagnosis and adequate therapeutic orientation.(AU)

Urticaria papular y sus agentes causales en Colombia / Papular urticaria: A review of causal agents in Colombia

Resumen La urticaria papular es una enfermedad alérgica causada por la picadura de insectos, la cual predomina en el trópico. El objetivo de esta revisión fue profundizar en sus aspectos epidemiológicos e inmunológicos, particularmente con base en datos publicados en Latinoamérica. Se hizo una revisión no sistemática mediante la búsqueda electrónica de artículos sobre la epidemiología de la urticaria papular, las características entomológicas de los agentes causales y los mecanismos inmunológicos asociados. Según los diversos reportes de centros médicos de Latinoamérica la urticaria papular es frecuente; el único estudio de prevalencia publicado indica que afecta a una cuarta parte de los niños escolares de Bogotá. Hay información sobre la relación causal entre la exposición domiciliaria a la pulga, la pobreza y la urticaria papular en Bogotá, una ciudad representativa de las altitudes andinas. No hay estudios que indaguen directamente sobre los insectos causales en zonas cálidas, aunque se sospecha clínicamente de los mosquitos Aedes aegypti y Culex quinquefasciatus. En cuanto a su patogenia, se destaca la participación de mecanismos celulares que involucran las células colaboradoras Th2, lo cual explica que sea una condición de hipersensibilidad retardada. El papel de la inmunoglobulina E (IgE) en la urticaria papular no está tan claro. Se desconocen los antígenos derivados de los insectos que causan la enfermedad, aunque se plantea que existen moléculas comunes de reacción cruzada entre los insectos, tales como el alérgeno Cte f 2 en la pulga, y sus homólogos en los mosquitos. La urticaria papular es una condición frecuente en Latinoamérica que debe investigarse en profundidad. La caracterización inmunológica de los componentes moleculares que causan esta condición puede resolver interrogantes sobre su etiología y su patogenia.

Abstract Papular urticaria is a chronic allergic reaction induced by insect bites, which is common in the tropics. The objective of this review was to deepen on epidemiological and immunological aspects of this disease, focused on data published in Latin American countries. We conducted a non-systematic review of the literature through electronic search on the epidemiology of papular urticaria, the entomological characteristics of the causative agents and associated immunological mechanisms. Several reports from medical centers suggest that papular urticaria is common in Latin America. Only one epidemiological survey designed to estimate prevalence of papular urticaria has been published, reporting that about a quarter of children under six years of age is affected by this condition in Bogotá. There is evidence on the causal relationship among exposure to indoor fleas, poverty and papular urticaria in Bogotá, a representative city of the Andean altitudes. Information about causal insects in tropical warmer areas is scarce, although from clinical reports Aedes aegypti and Culex quienquefasciatus appear to be the most common. Th2 cellular-mediated mechanisms are involved in its pathogenesis, which explains its delayed hypersensitivity. The role of immunoglobulin E is not clear in this disease. Insect-derived antigens directly involved in papular urticaria etiology are unknown. However, it is possible that common molecules among causal insects mediate cross-reactive reactions, such as Cte f 2 allergen, found in cat fleas, and its counterparts in mosquitoes. Papular urticaria is a frequent disease in Latin America that should be further investigated. Immunological characterization of the molecular components that cause this condition may solve questions about its pathogenesis.

Cloning and expression of recombinant equine interleukin-3 and its effect on sulfidoleukotriene and cytokine production by equine peripheral blood leukocytes.

Interleukin-3 is a growth and differentiation factor for various hematopoietic cells. IL-3 also enhances stimulus-dependent release of mediators and cytokine production by mature basophils. Function of IL-3 has not been studied in horses because of lack of horse-specific reagents. Our aim was to produce recombinant equine IL-3 and test its effect on sulfidoleukotriene and cytokine production by equine peripheral blood leukocytes (PBL). Equine IL-3 was cloned, expressed in E. coli and purified. PBL of 19 healthy and 20 insect bite hypersensitivity (IBH)-affected horses were stimulated with Culicoides nubeculosus extract with or without IL-3. Sulfidoleukotriene (sLT) production was measured in supernatants by ELISA and mRNA expression of IL-4, IL-13 and thymic stromal lymphopoietin (TSLP) assessed in cell lysate by quantitative real-time PCR. Recombinant equine IL-3 (req-IL-3) had a dose dependent effect on sLT production by stimulated equine PBL and significantly increased IL-4, IL-13 and TSLP expression compared to non-primed cells. IL-3 priming significantly increased Culicoides-induced sLT production in IBH-affected but not in non-affected horses and was particularly effective in young IBH-affected horses (≤ 3 years). A functionally active recombinant equine IL-3 has been produced which will be useful for future immunological studies in horses. It will also allow improving the sensitivity of cellular in vitro tests for allergy diagnosis in horses.

Detection of feline herpes virus 1 via polymerase chain reaction and immunohistochemistry in cats with ulcerative facial dermatitis, eosinophilic granuloma complex reaction patterns and mosquito bite hypersensitivity.

Ulcerative dermatitis caused by feline herpes virus 1 (FHV-1) is an uncommon disease characterized by cutaneous ulcers secondary to epidermal, adnexal and dermal necrosis. Differential diagnoses for FHV-1 lesions include, but are not limited to, mosquito bite hypersensitivity and eosinophilic granuloma complex. Histopathological diagnosis of FHV-1 dermatitis is based on the detection of the intranuclear inclusion bodies. In cases where intranuclear inclusions are missing but clinical and histological findings are compatible with FHV-1 dermatitis, immunohistochemistry (IHC) and PCRs have been used. In this retrospective study, we evaluated the presence of FHV-1 by IHC and PCR in skin biopsies and compared the results of the two tests. Sixty-four skin biopsy specimens from cats with compatible lesions were reviewed and tested via PCR and IHC for evidence of FHV-1. Polymerase chain reaction was positive in 12 of 64 biopsies; PCR and IHC were positive only in two of 64 biopsies, and these cases were considered true positive cases. The higher number of PCR-positive cases was possibly attributed to amplification of viral DNA from a live attenuated vaccination, but a previous FHV-1 infection with subsequent amplification of latently inserted FHV-1 could not be excluded. If clinical signs and histopathology suggest FHV-1 infection in the absence of typical inclusion bodies, IHC is the preferred diagnostic test; PCR may be useful for initial screening, but due to false positives is not sufficient for a definitive diagnosis.

Skin-infiltrating T cells and cytokine expression in Icelandic horses affected with insect bite hypersensitivity: a possible role for regulatory T cells.

Equine insect bite hypersensitivity (IBH) is a seasonally recurrent, pruritic skin disorder caused by an IgE-mediated reaction to salivary proteins of biting flies, predominantly of the genus Culicoides. The aim of this study was to define T cell subsets and cytokine profile in the skin of IBH-affected Icelandic horses with particular focus on the balance between T helper (Th) 1, Th2 and T regulatory (Treg) cells. Distribution and number of CD4+, CD8+ and Forkhead box P3 (FoxP3)+ T cells were characterized by immunohistochemical staining in lesional and non-lesional skin of moderately and severely IBH-affected horses (n=14) and in the skin of healthy control horses (n=10). Using real-time quantitative reverse transcription-polymerase chain reaction, mRNA expression levels of Th2 cytokines (Interleukin (IL)-4, IL-5, IL-13), Th1 cytokines (Interferon-γ), regulatory cytokines (Transforming Growth Factor ß1, IL-10) and the Treg transcription factor FoxP3 were measured in skin and blood samples. Furthermore, Culicoides nubeculosus specific serum IgE levels were assessed. Lesions of IBH-affected horses contained significantly higher numbers of CD4+ cells than skin of healthy control horses. Furthermore, the total number of T cells (CD4+ and CD8+) was significantly increased in lesional compared to non-lesional skin and there was a tendency (p=0.07) for higher numbers of CD4+ cells in lesional compared to non-lesional skin. While the number of FoxP3+ T cells did not differ significantly between the groups, the ratio of Foxp3 to CD4+ cells was significantly lower in lesions of severely IBH-affected horses than in moderately affected or control horses. Interestingly, differences in FoxP3 expression were more striking at the mRNA level. FoxP3 mRNA levels were significantly reduced in lesional skin, compared both to non-lesional and to healthy skin and were also significantly lower in non-lesional compared to healthy skin. Expression levels of IL-13, but not IL-4 or IL-5, were significantly elevated in lesional and non-lesional skin of IBH-affected horses. IL-10 levels were lower in lesional compared to non-lesional skin (p=0.06) and also lower (p=0.06) in the blood of IBH-affected than of healthy horses. No significant changes were observed regarding blood expression levels of Th1 and Th2 cytokines or FoxP3. Finally, IBH-affected horses had significantly higher Culicoides nubeculosus specific serum IgE levels than control horses. The presented data suggest that an imbalance between Th2 and Treg cells is a characteristic feature in IBH. Treatment strategies for IBH should thus aim at restoring the balance between Th2 and Treg cells.

The effect of a supplement containing sunflower oil, vitamins, amino acids, and peptides on the severity of symptoms in horses suffering insect bite hypersensitivity.

Insect bite hypersensitivity (IBH) is most often caused by Culicoides spp., although other insect species are also thought to play a role in causing this disease. The disease has been reported to occur in most countries, with a prevalence of up to 28% in some countries in some breeds. The only truly effective therapy currently available is to prevent horses from coming into contact with Culicoides midges, which can be achieved by stabling horses during certain times of the day or by covering them with insect blankets. The effect of a new dietary supplement containing vitamins, polypeptides, and amino acids on IBH severity was tested at the start of the Culicoides season (April 2009). Fifty horses participated in this placebo-controlled, double-blinded study: 25 horses received the supplement and 25 received placebo for 30 days; thereafter all horses received the supplement for a further 30 days. The investigators scored the severity of IBH in all horses, based on clinical evaluation and digital photographs taken before and after the first 30 days of the trial. The owners were asked to assess the severity of the signs prior to the investigation and after the first and second 30-day periods. For each period, IBH clinical symptoms were classified as increased, decreased, or the same. Results showed that investigator-assessed symptoms became worse in more horses receiving placebo than in horses receiving supplement, indicating a positive effect of the supplement. However, there were no treatment-group differences when symptom severity was scored by the horse owners. No side-effects were observed.

[Prevalence of hereditary diseases in three-year-old horses of the Freiberger breed]. / Erhebung der Prävalenz von Erbkrankheiten bei dreijährigen Pferden der Freiberger-Rasse.

The objective of this study was to investigate clinical signs indicating diseases with known or suspected hereditary components like equine sarcoid, insect bite hypersensitivity, osteochondrosis, allergic airway diseases, podotrochleosis, prognatism and wind-sucking in the franches-montagnes horse. We performed a clinical examination on 702 three-year-old, privately owned franches-montagnes horses, which were shown at the Swiss-Field-Tests in 2004. With the help of the owners a questionnaire on health, environment and feeding habits of the animals was completed. In 11.9% of the horses, sarcoids were detected, 6.2% carried one tumor and 5.7% had several. The prevalence was higher in chestnuts (16.6%) than in bays (10.1%). The prevalence of sarcoids in offspring from sires with known sarcoids (data from Station-Tests 1994-2005) was not significantly higher than in descendants from stallions without a known history of sarcoids. Clinical signs of insect bite hypersensitivity were only found in six horses (0.9%). In 12.0% hoof abnormalities like brittle horn, ring hoofs or hoof cracks of different degrees were recorded. The angle between hoof base and hoof wall was 56.7 +/- 0.1 degrees, the average hoof width was 13.7 +/- 0.3 cm in the front feet. For both of these measures we found no significant difference between left and right feet. With the exception of a high sarcoid prevalence, our results indicate that the franches-montagnes horse is a healthy breed overall.

Repellent effects of the essential oil of Lavendula angustifolia against adults of Hyalomma marginatum rufipes.

The repellent effects of the essential oil of Lavendula angustifolia on adults of Hyalomma marginatum rufipes was studied at concentrations of 5, 10 and 20% v/v. A suitable tick climbing bioassay based on the questing behaviour of ticks was used to test for repellency. High percentage repellency (range 70-100) was shown at all concentrations of the essential oil of L. angustifolia, although at 5% v/v it only persisted for the first 40 minutes compared with 120 minutes at other concentrations (10 and 20% v/v). The repellent strength of L. angustifolia compared well (P > 0.05) with that of DEET (N,N-diethyl-m-toluamide), a commercial reference repellent, for the 2-hour period of the study.

Onchocerca ochengi acquisition in zebu Gudali cattle exposed to natural transmission: parasite population dynamics and IgG antibody subclass responses to Ov10/Ov11 recombinant antigens.

Ngaoundere Gudali zebu cattle naturally exposed to Simulium damnosum s.l. and Culicoides spp. bites were examined during 4 years for O. ochengi adult worm acquisition, Onchocerca ochengi and Onchocerca gutturosa skin microfilaria dynamics, and IgG1 and IgG2 antibody subclass responses. Eleven animals acquired a total of 465 O. ochengi nodules (average of 17 per female and 72 per male). The O. ochengi nodule load was highly variable in individual animals and exacerbated in mature male cattle. Three patterns of acquisition of O. ochengi (resistant to new infestation, early susceptibility and late susceptibility), not associated with Simulium biting intensity (P > 0.05), were distinguished. The minimum prepatent periods for O. ochengi nodules, O. ochengi microfilariae and O. gutturosa microfilariae were 10, 20 and 21 months, respectively. The O. ochengi microfilaria density significantly (P < 0.001) increased with age, was higher in young mature bulls than female animals (P < 0.001) and finally reached highest levels (P < 0.005) during the dry season. Antibody responses to Ov10/Ov11 recombinant O. volvulus antigens were predominantly of the IgG1 subclass. High levels of this subclass (not IgG2) observed in new born calves declined to almost zero levels at the age of 5-8 months but IgG1 levels significantly increased (P < 0.05) with age subsequently during patency. Put together the acquisition and accumulation of O. ochengi parasites in zebu cattle, apart from being season, sex (gender) and host age associated, may also suggest a density-dependent regulation of parasite establishment in a proportion of the exposed population.

Flea bite hypersensitivity: new aspects on the involvement of mast cells.

A study was performed to test the effect of sensitization to flea antigen, followed by exposure to fleas on mast cells (MCs), their subtypes, and IgE+ cells. Biopsies were taken from flea-sensitized dogs (n=28) and non-sensitized dogs (n=5) that had been exposed to fleas. Control groups consisted of flea-sensitized (n=12) and non-sensitized dogs (n=9) that were not exposed to fleas. Biopsies, taken before, 24 and 72 h after local flea exposure, were stained with haematoxylin and eosin (H&E), toluidine blue, a double labelling technique for MC chymase and tryptase and anti-IgE. An intradermal test for flea antigen was performed and serum titres of allergen-specific IgE and IgG were measured. Significantly higher numbers (P<0.001) of double labelled MCs compared to toluidine blue stained MCs were detectable in flea-sensitized dogs independent of flea exposure. In contrast, in non-sensitized dogs, the number of toluidine blue stained MCs and the number of double labelled MCs did not differ. In flea-sensitized dogs after flea exposure the percentage of C-MC was significantly increased at day 1 (P<0.001) and day 3 (P<0.001), whereas the percentage of TC-MCs decreased significantly at day 1 (P<0.001) and day 3 (P<0.05). The percentage of T-MCs decreased (P<0.05 day 0 versus day 1; P<0.05 day 0 versus day 3). No significant difference was detectable after toluidine blue staining and staining for IgE+ cells between the groups nor between the MC density and the number of IgE+ cells. All flea-sensitized dogs had positive skin tests to flea antigen and high serum titres of flea-specific serum IgE and IgG antibodies. In non-sensitized dogs, these results were negative. Our data provide strong evidence for an upregulation of MC proteases during the process of sensitization and a generalized selective release of mast cell tryptase after exposure to the antigen.

Role of the chemokine eotaxin in the pathogenesis of equine sweet itch.

The chemokine eotaxin is involved in the recruitment of eosinophils and T helper 2 lymphocytes in human allergic diseases, and drugs that block its activity, including eotaxin receptor (CCR3) antagonists, are being developed. The authors have recently cloned the horse ortholog of eotaxin and shown that it can induce equine eosinophil migration and activation in vitro. Moreover, eotaxin mRNA expression was upregulated in cultured horse dermal fibroblasts exposed to equine interleukin-4, suggesting a possible source of this eosinophil chemoattractant in equine skin. The results of this study show that eotaxin and monocyte chemoattractant protein (MCP) 1, but not MCP-2 or MCP-4, mRNA expression is upregulated in skin biopsies of sweet itch lesions when eosinophils are present, when compared with clinically normal skin from the same ponies.

Immunoglobulin-E-bearing cells in skin biopsies of horses with insect bite hypersensitivity.

The aim of the present study was to investigate, with immunohistochemistry and in situ hybridisation, if immunoglobulin-E (IgE) and mast cells are involved in the pathogenesis of insect bite hypersensitivity (IBH), an allergic dermatitis of horses. In tissue sections fixed in paraformaldehyde (PFA) for <24 h, significantly more IgE protein-bearing cells were found in the dermis and epidermis of acute and chronic IBH lesions than in skin biopsies from healthy horses (medians = 466, 236 and 110 cells/mm2, respectively; P < or = 0.01). More IgE-mRNA positive (+) cells were observed in the dermis of acute IBH lesions than in the dermis of healthy skin (median = 2.8 vs. 0.0 cells/mm2; P < or = 0.01). Significantly, more mast cells were detected with metachromatic (median = 160 vs. 62 cells/mm2; P < or = 0.001) and tryptase-specific stainings (median = 120 vs. 69 cells/mm2; P < or = 0.001) in the dermis of acute IBH biopsies compared to healthy skin. No chymase+ mast cells were found in any skin biopsy. IBH lesions fixed in PFA for >24 h were compared to dermatomycosis (DM) lesions; IBH biopsies contained a similar number of IgE-protein+ cells to DM biopsies (median = 249 vs. 192 cells/mm2; P = 0.08) but had significantly more IgE-mRNA+, metachromatic and tryptase+ mast cells than DM biopsies. This study suggests an involvement of IgE-mediated immune reactions in the pathogenesis of IBH as well as, sometimes, in dermatomycosis. Using double labelling, cells which expressed IgE protein and contained mast cell enzymes were detected.