Culicoides obsoletus extract relevant for diagnostics of insect bite hypersensitivity in horses.
Abstract: Insect bite hypersensitivity (IBH) is an allergic dermatitis in horses caused by the bites of Culicoides species. The aim of the present study was to evaluate the applicability of whole body extracts of C. obsoletus (the main species found feeding on horses in the Netherlands), C. nubeculosus (rarely found in The Netherlands) and C. sonorensis (typical for North America) for diagnosis of IBH in horses in The Netherlands. Blood and serum samples of 10 clinically confirmed IBH affected and 10 healthy control horses were used to evaluate the IgE titers (ELISA) against the Culicoides whole body extracts of the three Culicoides species. Basophil degranulation was assessed by histamine release test (HRT) after stimulation with these extracts at 5, 0.5 and 0.05 μg/ml. IBH affected horses had significantly higher IgE titers against C. obsoletus than against C. nubeculosus and C. sonorensis. Furthermore, C. obsoletus induced significantly higher histamine release in whole blood of IBH affected horses compared to the other extracts at 0.5 μg/ml. Western blot data revealed IgE binding to many proteins in C. obsoletus extract. This interaction was absent or weak in C. nubeculosus and C. sonorensis extracts for IBH affected horses. Results on individual level indicate that the HRT is more sensitive than ELISA in diagnosing IBH. However, ELISA is more practical as a routine test, therefore the ELISA was further evaluated using C. obsoletus extract on 103 IBH affected and 100 healthy horses, which resulted in a test sensitivity and specificity of 93.2% and 90.0%, respectively. The IgE ELISA readings enabled the analysis of the predicted probability of being IBH affected. From an optical density 450nm value of 0.33 onwards, the probability of IBH affected was more than 0.9. The results presented in this paper show that the use of native Culicoides spp. that feed on horse, is important for improved diagnosis and that the described ELISA based on C. obsoletus can be used routinely to diagnose IBH in countries where this species is the main Culicoides feeding on horses.
Copyright © 2012 Elsevier B.V. All rights reserved.
Publication Date: 2012-07-23 PubMed ID: 22906994DOI: 10.1016/j.vetimm.2012.07.007Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research focused on using insect extracts to diagnose insect bite hypersensitivity (IBH) in horses, particularly extracts from the Culicoides species that frequently feed on horses in the Netherlands. The study concluded that the use of these native Culicoides extracts could improve IBH diagnosis and that the described ELISA test based on one species, C. obsoletus, could be used routinely for this diagnosis in countries where this insect species is the main Culicoides affecting horses.
Objective of the Research
- The main aim of the study was to determine the effectiveness of whole body extracts from different species of Culicoides for diagnosing insect bite hypersensitivity (IBH) in horses. Specifically, they tested extracts from C. obsoletus, C. nubeculosus, and C. sonorensis.
Methodology
- Blood and serum samples from 10 horses clinically confirmed to have IBH and 10 healthy control horses were used to evaluate the immunoglobulin E (IgE) titers against the three Culicoides species.
- The researchers used an ELISA test to measure the IgE titers and a histamine release test to assess basophil degranulation after stimulating the samples with the insect extracts.
Findings
- The ELISA tests revealed significantly higher IgE titers against C. obsoletus compared to the other two species in the IBH-affected horses.
- C. obsoletus also induced a much higher histamine release in whole blood samples from the affected horses. Comparatively, the other extracts had a less pronounced effect.
- Western blot results showed that IgE binds with many proteins in the C. obsoletus extract, suggesting a potential reason for the high IgE titers and histamine release.
- Results at the individual level implied that the histamine release test (HRT) was more sensitive in diagnosing IBH than the ELISA. However, due to practicality, the ELISA is deemed more suitable for routine testing.
- Further evaluation of the ELISA was performed using C. obsoletus extract on a larger sample of horses (103 IBH affected and 100 healthy), achieving high sensitivity and specificity for diagnosing IBH.
Conclusion
- The findings showed that using native Culicoides species that feed on horses is important to improve the diagnosis of IBH.
- The research also supports the routine use of an ELISA test based on C. obsoletus for diagnosing IBH in regions where this insect species predominantly feeds on horses.
Cite This Article
APA
van der Meide NM, Meulenbroeks C, van Altena C, Schurink A, Ducro BJ, Wagner B, Leibold W, Rohwer J, Jacobs F, van Oldruitenborgh-Oosterbaan MM, Savelkoul HF, Tijhaar E.
(2012).
Culicoides obsoletus extract relevant for diagnostics of insect bite hypersensitivity in horses.
Vet Immunol Immunopathol, 149(3-4), 245-254.
https://doi.org/10.1016/j.vetimm.2012.07.007 Publication
Researcher Affiliations
- Cell Biology and Immunology Group, Wageningen University, Wageningen, The Netherlands.
MeSH Terms
- Animals
- Ceratopogonidae / chemistry
- Ceratopogonidae / immunology
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / standards
- Enzyme-Linked Immunosorbent Assay / veterinary
- Female
- Histamine Release / immunology
- Horse Diseases / blood
- Horse Diseases / diagnosis
- Horse Diseases / immunology
- Horses
- Hypersensitivity / blood
- Hypersensitivity / immunology
- Hypersensitivity / veterinary
- Immunoglobulin E / blood
- Insect Bites and Stings / blood
- Insect Bites and Stings / diagnosis
- Insect Bites and Stings / immunology
- Insect Bites and Stings / veterinary
- Logistic Models
- Predictive Value of Tests
- Sensitivity and Specificity
Grant Funding
- BBS/E/I/00001445 / Biotechnology and Biological Sciences Research Council
- BBS/E/I/00001701 / Biotechnology and Biological Sciences Research Council
Citations
This article has been cited 10 times.- Pessoa VC, Branco-Ferreira M, Jónsdóttir S, Marti E, Tilley P. Comparison of Skin Prick Tests (SPT), Intradermal Tests (IDT) and In Vitro Tests in the Characterization of Insect Bite Hypersensitivity (IBH) in a Population of Lusitano Horses: Contribution for Future Implementation of SPT in IBH Diagnosis.. Animals (Basel) 2023 Aug 28;13(17).
- Cox A, Stewart AJ. Insect Bite Hypersensitivity in Horses: Causes, Diagnosis, Scoring and New Therapies.. Animals (Basel) 2023 Aug 4;13(15).
- Birras J, White SJ, Jonsdottir S, Novotny EN, Ziegler A, Wilson AD, Frey R, Torsteinsdottir S, Alcocer M, Marti E. First clinical expression of equine insect bite hypersensitivity is associated with co-sensitization to multiple Culicoides allergens.. PLoS One 2021;16(11):e0257819.
- Raza F, Babasyan S, Larson EM, Freer HS, Schnabel CL, Wagner B. Peripheral blood basophils are the main source for early interleukin-4 secretion upon in vitro stimulation with Culicoides allergen in allergic horses.. PLoS One 2021;16(5):e0252243.
- Novotny EN, White SJ, Wilson AD, Stefánsdóttir SB, Tijhaar E, Jonsdóttir S, Frey R, Reiche D, Rose H, Rhyner C, Schüpbach-Regula G, Torsteinsdóttir S, Alcocer M, Marti E. Component-resolved microarray analysis of IgE sensitization profiles to Culicoides recombinant allergens in horses with insect bite hypersensitivity.. Allergy 2021 Apr;76(4):1147-1157.
- Raza F, Ivanek R, Freer H, Reiche D, Rose H, Torsteinsdóttir S, Svansson V, Björnsdóttir S, Wagner B. Cul o 2 specific IgG3/5 antibodies predicted Culicoides hypersensitivity in a group imported Icelandic horses.. BMC Vet Res 2020 Aug 10;16(1):283.
- François L, Hoskens H, Velie BD, Stinckens A, Tinel S, Lamberigts C, Peeters L, Savelkoul HFJ, Tijhaar E, Lindgren G, Janssens S, Ducro BJ, Buys N, Schurink AA. Genomic Regions Associated with IgE Levels against Culicoides spp. Antigens in Three Horse Breeds.. Genes (Basel) 2019 Aug 8;10(8).
- Lavergne SN. In Vitro Research Tools in the Field of Human Immediate Drug Hypersensitivity and Their Present Use in Small Animal Veterinary Medicine.. Vet Sci 2016 Dec 22;4(1).
- Mueller RS, Janda J, Jensen-Jarolim E, Rhyner C, Marti E. Allergens in veterinary medicine.. Allergy 2016 Jan;71(1):27-35.
- Meulenbroeks C, van der Lugt JJ, van der Meide NM, Willemse T, Rutten VP, Zaiss DM. Allergen-Specific Cytokine Polarization Protects Shetland Ponies against Culicoides obsoletus-Induced Insect Bite Hypersensitivity.. PLoS One 2015;10(4):e0122090.
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