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NCUR Proceedings NCUR 2021 Proceedings Amphibian Skin Pathogen Batrachochytrium dendrobatidis Surveillance in Ecuador Ecology - Time: Wed 1:30pm-2:30pm - Session Number: 1015 Connor Chapman, Abby Piddock, Christian Sorenson, Jaden Seman, Nathan Lamb, Kirsten Enns, Kyle Harris, Matthew Becker, Sheryl Welfel, Alejandro Arteaga. Department of Biology and Chemistry, Liberty University.1971 University Blvd, Lynchburg, VA 24515. Connor Chapman Background: In recent decades, amphibians have experienced unprecedented population declines, leading to species extinctions worldwide. Chytridiomycosis has been linked to these declines and is caused by the fungal pathogen, Batrachohytrium dendrobatidis (Bd). In relation to South America, this disease is putting many amphibian species within Ecuador and surrounding areas at risk of extinction. This disease survey will build upon existing data to determine if Bd is persisting in the Ecuadorian upper Amazon and the intensity of infection for sampled amphibians. This pathogen surveillance will aid the ongoing global conservation efforts by documenting amphibians afflicted by this waterborne fungus. Research Methods: Our survey site was Jatun Sacha Biological Research Stations near Tena, Ecuador. Students and faculty completed amphibian surveys in the surrounding jungle. 30 Amphibians were sampled opportunistically. To sample potential pathogens, the skin of each amphibian was swabbed with a sterile rayon swab by streaking the swab across the ventral surface 10 times, each lateral surface 5 times, and each hind foot 5 times for a total 30 times. Swabs were then placed in sterile 1.5 ml microcentrifuge tubes and stored for return to the laboratory. Samples are in the process of being exported from Ecuador. Once received, individual samples will have DNA extracted from swabs using the Qiagen DNeasy Blood & Tissue Kit according to the manufacturer’s protocol. DNA obtained from the swabs will be amplified with pathogen specific real-time PCR (qPCR) assays to quantify Bd. All samples will initially be amplified in singlicate. Any sample that returns a positive result will be assayed again in duplicate to confirm the initial result and accurately estimate infection intensity. Results/findings: This research is ongoing, and the qPCR results should be obtained within the next few weeks. A total of 30 amphibians (15 canopy dwelling species and 15 forest floor dwelling species) have been properly swabbed and the swabs have been stored in tubes (with DNA Safe) and are ready to be mailed back from Quito, Ecuador (as of February 1st, 2021). Once the swabs are received, DNA will be extracted, and the qPCR protocol will be conducted to determine any presence of Bd. Based upon the elevation and prevailing warmer temperatures of the upper Amazon collection site, we do not anticipate to find the presence of BD within our samples. We do anticipate to find positive Bd results when we sample in the Cloud forest next year. Conclusions: We do not expect to find Bd in our current samples from the upper Amazon due to the higher temperatures, but we do expect to find Bd when we sample at higher elevations in coming years. In conclusion, this pathogen surveillance will aid the ongoing global conservation efforts by documenting amphibians afflicted by this waterborne fungus. Implications for further research: If amphibians are found to be healthy and infected with Bd, we plan to also assess the overall skin microbiome to see if there is some degree of resistance to the skin pathogen. In the coming years, we plan to continue surveying at different locations in Ecuador to further our understanding of the presence of Bd in this area and will aid in long-term conservation. Background: In recent decades, amphibians have experienced unprecedented population declines, leading to species extinctions worldwide. Chytridiomycosis has been linked to these declines and is caused by the fungal pathogen, Batrachohytrium dendrobatidis (Bd). In relation to South America, this disease is putting many amphibian species within Ecuador and surrounding areas at risk of extinction. This disease survey will build upon existing data to determine if Bd is persisting in the Ecuadorian upper Amazon and the intensity of infection for sampled amphibians. This pathogen surveillance will aid the ongoing global conservation efforts by documenting amphibians afflicted by this waterborne fungus. Research Methods: Our survey site was Jatun Sacha Biological Research Stations near Tena, Ecuador. Students and faculty completed amphibian surveys in the surrounding jungle. 30 Amphibians were sampled opportunistically. To sample potential pathogens, the skin of each amphibian was swabbed with a sterile rayon swab by streaking the swab across the ventral surface 10 times, each lateral surface 5 times, and each hind foot 5 times for a total 30 times. Swabs were then placed in sterile 1.5 ml microcentrifuge tubes and stored for return to the laboratory. Samples are in the process of being exported from Ecuador. Once received, individual samples will have DNA extracted from swabs using the Qiagen DNeasy Blood & Tissue Kit according to the manufacturer’s protocol. DNA obtained from the swabs will be amplified with pathogen specific real-time PCR (qPCR) assays to quantify Bd. All samples will initially be amplified in singlicate. Any sample that returns a positive result will be assayed again in duplicate to confirm the initial result and accurately estimate infection intensity. Results/findings: This research is ongoing, and the qPCR results should be obtained within the next few weeks. A total of 30 amphibians (15 canopy dwelling species and 15 forest floor dwelling species) have been properly swabbed and the swabs have been stored in tubes (with DNA Safe) and are ready to be mailed back from Quito, Ecuador (as of February 1st, 2021). Once the swabs are received, DNA will be extracted, and the qPCR protocol will be conducted to determine any presence of Bd. Based upon the elevation and prevailing warmer temperatures of the upper Amazon collection site, we do not anticipate to find the presence of BD within our samples. We do anticipate to find positive Bd results when we sample in the Cloud forest next year. Conclusions: We do not expect to find Bd in our current samples from the upper Amazon due to the higher temperatures, but we do expect to find Bd when we sample at higher elevations in coming years. In conclusion, this pathogen surveillance will aid the ongoing global conservation efforts by documenting amphibians afflicted by this waterborne fungus. Implications for further research: If amphibians are found to be healthy and infected with Bd, we plan to also assess the overall skin microbiome to see if there is some degree of resistance to the skin pathogen. In the coming years, we plan to continue surveying at different locations in Ecuador to further our understanding of the presence of Bd in this area and will aid in long-term conservation. An Analysis of Florida Fighting Conch Tissue for the Presence of Microplastics Ecology - Time: Tue 12:30pm-1:30pm - Session Number: 4106 Jordan Kleinschmidt, Dr. Alexis Janosik, Department of Biology, University of West Florida, 11000 University Pkwy, Pensacola, FL 32514 Jordan Kleinschmidt Microplastics are defined as small pieces of plastic that measure five millimeters or less in size and come in many shapes and colors. These miniscule pieces of debris can be found all throughout the oceans, lakes, and can even be found inside organisms. The source of these microplastics can be from beauty products, break down of larger plastic debris, and other types of microbeads and bits of plastic from manufactured goods. There are many dangers posed by microplastic pollution, one being the consumption of these debris by marine organisms, which will later be consumed by humans. The effects of consuming microplastics on marine organisms is actively being researched however, the consumption and thus the effects of microplastics on organisms is still not fully known. The Florida Fighting Conch is a gastropod mollusc that can be found from the beaches of North Carolina all the way down to Florida in the Gulf of Mexico. These conchs are commonly found washed up onto the shores of beaches due to heavy current and wave activity. Florida Fighting Conchs serve as a food source for other organisms in the ecosystem. As such, the potential for biomagnification up the food chain is present if Florida Fighting Conchs are consuming microplastics. Knowing that these conchs are feeding off of algae in the seagrass beds, they have the potential to also consume benthic microplastics. The objective of this study is to quantify microplastic contamination in Florida Fighting Conchs. Florida Fighting conchs will be collected locally from intertidal beach habitats and tissue will be digested for microplastic quantification. After tissue digestion is complete, samples will be filtered and microplastics will be quantified and measured using microscopy. Microplastics are defined as small pieces of plastic that measure five millimeters or less in size and come in many shapes and colors. These miniscule pieces of debris can be found all throughout the oceans, lakes, and can even be found inside organisms. The source of these microplastics can be from beauty products, break down of larger plastic debris, and other types of microbeads and bits of plastic from manufactured goods. There are many dangers posed by microplastic pollution, one being the consumption of these debris by marine organisms, which will later be consumed by humans. The effects of consuming microplastics on marine organisms is actively being researched however, the consumption and thus the effects of microplastics on organisms is still not fully known. The Florida Fighting Conch is a gastropod mollusc that can be found from the beaches of North Carolina all the way down to Florida in the Gulf of Mexico. These conchs are commonly found washed up onto the shores of beaches due to heavy current and wave activity.
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