Building a DNA library for isopods and their viral pathogens

$416
Raised of $2,050 Goal
21%
Ended on 11/21/19
Campaign Ended
  • $416
    pledged
  • 21%
    funded
  • Finished
    on 11/21/19

About This Project

Iridoviruses have caused large-scale mortality due to systemic disease, threatening the environment by jeopardizing many species. How do these viruses spread? We hypothesize that iridoviruses spread through cannibalism and injection. A comprehensive study of these viruses in isopods will help us understand isopod ecology and biology, as well as add to our understanding of the virus family Iridoviridae. This information will be applied in creating an Isopod DNA Library.

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What is the context of this research?

The family Iridoviridae is part of a monophyletic group of viruses called nucleocytoplasmic large DNA viruses. Iridoviridae are known to infect fish, reptiles, amphibians, and arthropods, including at least 175 species across 52 families of ectothermic vertebrates, and contribute to population decline in all of these groups by causing mass mortality. This population decline is a serious threat to biodiversity. We hypothesize that iridoviruses spread through cannibalism and injection. The isopods to be observed include Porcellio laevis, Armadillidium vulgare, and Trachelipus rathkii. We will also create an Isopod DNA Library, inspired by work we completed during the previous school year, which included uploading a duckweed ferredoxin gene to the NCBI.

What is the significance of this project?

An understanding of how these viruses spread can apply to much more than just an attempt to rescue isopods from population decline. This can serve as a model for the transmission methods of other viruses in the Iridoviridae family that infect vertebrates, which are threatened by pathogens, deforestation, and climate change. It may also serve as a model for outbreaks in humans. There is a lack of research into iridoviruses, particularly in understanding infection methods and species-specificity (to be analyzed via barcoding). An isopod DNA library of the DNA of isopods and their pathogens does not exist, but could aid researchers in identifying isopod species, genes, or iridovirus strains. A large collection of isopod genetic material may reveal cryptic species and genetic diversity.

What are the goals of the project?

The first step in this project is to achieve an iridovirus infection (infected = purple or blue isopod) in a population of isopods. To do so, spreading methods--cannibalism, environmental exposure, and injection-- will be tested in three isopod species. Whether or not an infection method succeeds, as well as the fastest infection method, will be determined. The next step is to sequence isopod and viral DNA to evaluate the viruses' species-specificity and identify the viruses infecting the isopods. This includes genetically determining if the infected isopods are the same species by sequencing their COI and 18S genes. The last step includes constructing an online database and uploading the sequences of isopod and iridovirus genes.

Budget

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In order to care for the isopods long term, materials such as food and habitats are crucial. We are expecting to care for 500-1000 isopods for two years (how much time we have left in high school). Of these isopods, we will be infecting 540, 180 of each of the three species listed above (20 isopods for each of the three infection methods, for 3 trials, for three species=540). These isopods, as well as more from other species and various locations, will have their COI and 18S genes sequenced for the DNA library. Infection tools such as needles, as well as sterilization equipment, are needed for the infection procedure. A specific iridovirus primer, COI primer, and 18S primer, as well as DNA purification kits, are necessary for proceeding with the isopod DNA library, for which sequencing is required as well. We plan to purify 100 preps of DNA and sequence 50 sequences (this includes both isopod (18S and COI genes) and viral DNA (full genome) sequences).

Endorsed by

This experiment has real potential to make new discoveries in virology, ecology, conservation, and evolution. The student researchers are very diligent, insightful, and hardworking, which guarantees that this project will be completed on time and on budget. I can be certain about this statement because I have taught both of them, one in scientific research and one in accelerated chemistry during their sophomore years.

Flag iconProject Timeline

By Nov. 30, 2019, we hope to have achieved an iridovirus infection (at least one purple/blue isopod). At this point in the project, the goal would be just to achieve an infection by any of the infection methods we listed above.

The next phase of comparing the speed and success of different infection methods would then begin.

Sequencing, analysis, and database construction are set for 2020.

The latter milestones may be subject to rearrangement, as issues may arise with earlier steps.

Oct 07, 2019

Project Launched

Nov 30, 2019

Achieve iridovirus outbreak in an isopod population via any transmission method by given date

Dec 02, 2019

Update backers on Experiment, Compare speed of transmission methods--begin 3 trials

Mar 05, 2020

Complete 3 trials by given date

Mar 11, 2020

Share trial results on Experiment

Meet the Team

Moriah Eley
Moriah Eley
Thomas Gatzke
Thomas Gatzke

Moriah Eley

Hi! I am a high school junior and am working on my first research project.

In the first year of our three-year research program we each conducted PCR, gel electrophoresis, and restriction digest on duckweed genes, the sequences of which we later uploaded to the NCBI database. We also completed cell fractionation and a Western Blot analysis.

Before and during these experiments, I solidified plans to study how iridoviruses spread between individuals, but afterwards I also became interested in constructing an isopod-specific gene database that could house isopod genes from all different species and habitats, as well as the genes of their pathogens, such as iridoviruses.

Now that I am entering my second year of the program, I have the opportunity to conduct my own project-- something that could greatly impact and jumpstart my career as a research scientist.

Any and all funding would mean a lot to us. Thank you for your support!

Thomas Gatzke

Hello, I am a a junior this year at Princeton High School.

During freshman and sophomore years I did independent research on terrestrial isopod behavior. I observed the interactions of Porcellio scaber to see if any specific isopods where found with the same others day after day. In this way I hoped to identify whether or not isopods formed any sort of social connections or "friendships."

This year I have joined Moriah on her project, as a fellow isopod enthusiast. I hope that this project will be a launchpad for future research projects for myself, both in high school and beyond.

Many thanks for both your interest and your support!

Lab Notes

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Project Backers

  • 16Backers
  • 21%Funded
  • $416Total Donations
  • $26.00Average Donation
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