Date of Award

Summer 2016

Degree Type

Thesis

Degree Name

Master of Science (MS)

Department

Biology

First Advisor

Dr. Ralph Saporito

Abstract

Amphibians produce the majority of their defensive chemicals, however alkaloid defenses in poison frogs are sequestered from dietary arthropods. Alkaloids function as a defense against predators, and certain types appear to inhibit microbial growth. However, alkaloid defenses vary considerably among populations of poison frogs, reflecting geographic differences in availability of dietary arthropods. Consequently, environmentally driven differences in poison frog alkaloid defenses may have significant implications regarding their protection against pathogens. While natural alkaloid mixtures in poison frogs have recently been shown to inhibit growth of non-pathogenic microbes, no studies have examined the effectiveness of alkaloids against microbes that infect frogs. Herein, I examined how alkaloid defenses in the strawberry poison frog, Oophaga pumilio, affect growth of the known anuran pathogens Aeromonas hydrophila and Klebsiella pneumoniae. Frogs were collected from five locations throughout Costa Rica that are known to vary in their alkaloid profiles. Alkaloids were isolated from individual skins, and extracts were assayed against both pathogens. Microbe subcultures were inoculated with extracted alkaloids to create dose-response curves. Subsequent spectrophotometry and cell counting assays were used to assess growth inhibition. GC-MS was used to characterize and quantify alkaloids in frog extracts, and my results suggest that variation in alkaloid defenses lead to differences in inhibition of these pathogens. This study provides the first evidence that alkaloid variation in a dendrobatid poison frog is associated with differences in inhibition of anuran pathogens, and offers further support that alkaloid defenses in poison frogs confer protection against both pathogens and predators.

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