Date on Honors Thesis

Spring 4-9-2021

Major

Agriculture: Pre-Veterinary Medicine

Minor

Chemistry

Examining Committee Member

Kevin M. Miller, PhD, Co-Advisor

Examining Committee Member

Laura K. Hoffman, DVM, Co-Advisor

Examining Committee Member

Chris Trzepacz, PhD, Committee Member

Examining Committee Member

William DeWees, MS, DVM, Committee Member

Abstract/Description

Dog saliva contains a large variety of proteins, each with specific functions throughout the body. While some aid in digestion, others provide immune support for the body. In the case of human dog allergies, dog skin dander has been proven to cause allergic reactions to those who are sensitive to the allergens in the dogs. However, past studies have shown a difference in allergic reaction to specific breeds, breaking dog breeds into two main categories: hypoallergenic and non-hypoallergenic. Within these groups, there are different hypoallergenicity levels, but overall, the hypoallergenic dog group does not cause an allergic reaction, whereas non-hypoallergenic, or shedding dogs, do. Because all dogs in both groups shed dander from their skin, the allergic reaction must be catalyzed by a different physiological substance, one that can be potentially paired with the dogs’ shedding or lack thereof. The Can f 1 allergen, the most abundant allergen in canines, is suspected to initiate allergic reactions in humans. In this study, the allergens specific to the saliva in each dog group were compared via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot tests, and enzyme-linked immunosorbent assay (ELISA). In preliminary testing using SDS-PAGE and Western blot technology, the saliva samples were homogenized in a buffer solution and, with the use of an electric current, run through a gel made to separate specific proteins. The Western blot test pins a specific protein or allergen needed with the use of the corresponding allergen’s antibody. To quantify the concentration of the Can f 1 allergen in each sample, the Can f 1 allergen was isolated from dogs in each group and quantified to indicate the presence of the allergen in the saliva via ELISA. In these tests, each saliva sample was placed in the microtiter plate in individual wells and serially diluted. After the addition of a primary and secondary antibody, the amount of the Can f 1 allergen in each sample was quantified and comparatively analyzed to saliva samples in both dog groups. The testing process was hypothesized to yield results of a higher Can f 1 allergen in the saliva of dogs in the shedding group, and lower Can f 1 allergen in the saliva of dogs in the hypoallergenic group. The results displayed inconclusive evidence regarding the correlation between the Can f 1 allergen concentration in dog saliva to human allergies due to the small sample pool and high variability in dog breeds. With these results, further studies can be conducted to neaten the testing process and advance the current treatment for human allergies or prevent reactions via combating the source.

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