Identifying bacterial pathogens in natural Drosophila populations

Kailene Richbow Dozier, a junior at Virginia Union University, worked with Dr. Kelly Dyer to examine the pathogens present in wild populations of Drosophila.

Abstract: An estimated 70% of the world’s insects are infected with the gram-negative bacteria, Wolbachia pipientis. Wolbachia is known to protect organisms against viral pathogens, however little is known about how it interacts with bacterial pathogens. The aim of this study was to isolate and identify bacterial pathogens within wild, mushroom feeding Drosophila. With this knowledge, we hope that we can aid future research studies trying to understand how Wolbachia increases host resistance to bacterial pathogens as compared to viral pathogens. Two species of flies, Drosophila Putrida and Drosophila Tripunctata were captured from Oconee State Park located in Athens, GA. The captured flies were separated by gender. Female flies were put aside to generate Iso-Female lines, while male flies were homogenized and streaked onto LB agar plates for bacteria growth. Once colonies began to grow after incubation, they were isolated and morphologically identified. 35 bacterial isolates were obtained from the 18 D. Tripunctata flies streaked. 20 bacterial isolates were obtained from the 20 D. Putrida flies streaked. A total of nine bacterial species were identified morphologically, seven of which were shared between the two species of wild flies. Similarities between the abundance of Micrococcus luteus and Enterococcus within both species of flies were observed. D. Tripunctata was seen to slightly differ from D. Putrida by carrying the bacterium Bacillus subtilis and Bacillus megaterium. For future exploration, 16S gene sequencing is needed to accurately confirm the identification of the bacterial pathogens discovered. Afterwards, testing for pathogenicity of the bacterial pathogens needs to be conducted as well as testing for Wolbachia’s interaction with the pathogens.

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The Battle of the Sperm: Observations of Sperm Competition in a Wolbachia Infected Species

LaTrice Montgomery, a student from Hampton University, worked with Paul Ginsberg in the lab of Dr. Kelly Dyer to study sperm competition in an infected fly species.

Abstract:  Wolbachia, a maternally inherited bacterium, is broadly distributed among arthropod hosts. It is also capable of reducing viral load in its host, preventing the transmission of human pathogens and thus raising the possibility of its use as a biocontrol agent. In this study, we ask whether Wolbachia has any effects on the mating system of its host, specifically whether it affects patterns of male fertility and/or sperm competition. We conducted mating trials using the fly, Drosophila recens, which is naturally infected with Wolbachia. We utilized an infected, dark eye recessive mutant D. recens and uninfected, red eye wild type D. recens to assess paternity of infected and uninfected males. We found that there are no advantages or disadvantages of Wolbachia infection for male fertility or sperm competition.

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Tricks not treats: The genetic basis of how a parasite manipulates its host’s reproduction

Wolbachia is a bacteria that is found in up to 60% of all insects, which is transmitted exclusively from mother to offspring through the egg. In many host species Wolbachia infection does not benefit the host, and instead  use a variety of tricks to manipulate its host’s reproduction in order to ensure its transmission to the next generation. This project investigates the interaction of Wolbachia and two of its hosts, the flies Drosophila recens and D. subquinaria. The same strain of Wolbachia manipulates each host differently: in D. recens Wolbachia causes the death of all the offspring when an infected male mates with an uninfected female, while the same strain of Wolbachia causes the death of sons of infected female D. subquinaria. We will ask how the genetic background of the host affects its interaction with Wolbachia, and then how this might lead to differences in the Wolbachia dynamics seen in the wild. This project is largely empirical, and will involve Drosophila culturing and crosses and molecular methods.

Host Laboratory: Kelly Dyer
Type of project: Empirical/laboratory-based

Geographic variation of Wolbachia-induced cytoplasmic incompatibility in the fly Drosophila recens

Sydney Keane, a Biology and Chemistry major from East Texas Baptist University, worked with Dr. Kelly Dyer examining the effects of infection on reproduction in Drosophila.

Abstract:  are bacterial parasites that commonly infect arthropods and nematodes. These parasites have damaging effects on the progeny of those they infect, including cytoplasmic incompatibility (CI). CI occurs when an infected male and an uninfected female mate, resulting in fewer eggs that successfully hatch into larvae than normal. In this study, infected virgin males from Drosophila recens were collected from multiple strains across three locations, and coupled with uninfected virgin females from the same species. After allowing the females to lay eggs for 72 hours, I recorded the numbers of eggs that hatched and that did not hatch. Males were tested for Wolbachia infection using PCR. After analyzing the data, I found that the overall hatch rate in each location was low, the amount of CI in each location did not vary significantly, the amount of CI in the experimental group compared to the control was significantly high, and that the number of total eggs produced varied significantly between the locations. The overall percentage of CI found within all of the locations examined was approximately 72%. These results show that the presence of Wolbachia is similarly effecting various populations of the fly throughout North America and that the level of CI occurring within this species may cause a drastic decrease in the population size over time.

 

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Is the Transmission Rate of The Wolbachia Parasite Lower in Hybrids Compared To Pure Species?

Jasmine Gipson, from Kennesaw State University, worked with Dr. Kelly Dyer in the UGA Genetics department to study the transmission of Wolbachia, a parasite of insects.

Abstract: Wolbachia is an endosymbiont parasite that lives in the reproductive system 70% of all insects. It is passed down vertically to its offspring from the mother. Wolbachia occurs naturally in D. recens, but not D. subquinaria. In the wild, D. recens and D. subquinaria hybridize and about 2-3% of those hybrid offsprings contain a D. recenâ’s mitochondria, but not the wolbachia. This is a strange situation because the mitochondria is only passed down through the mother’s eggs, just like wolbachia. So how is it possible to have a D. recenâ’s mitochondria, but not the wolbachia as well? This peculiar scenario led to the question, is the transmission rate of wolbachia lower in hybrids compared to pure species? To answer this question, a D. recens female was crossed with a D. subquinaria. The F1 hybrid female was then backcrossed to a D. subquinaria male. The F1 and F2 generations were both test for wolbachia using PCR. The transmission rate for the F1 generation had a transmission rate of 100% and the F2 generation had a transmission rate of 95.7%. This shows that the transmission rate of wolbachia is lowered in hybrid species compared to pure species. Possible explanations for this decrease in transmission rate is because of the genetic variation between D. recens and D. subquinaria or the parasite could have been randomly loss due to oogenesis.

 

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