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Immunological Responses to the White-Nose Syndrome Pathogen and their Potential Use as Control

Description

White-nose syndrome (WNS) is a fungal infection devastating bat populations throughout eastern North America. WNS is caused by a fungus, Pseudogymnoascus destructans (Pd), that invades the skin of hibernating bats. While there are a number of treatments being researched, there

White-nose syndrome (WNS) is a fungal infection devastating bat populations throughout eastern North America. WNS is caused by a fungus, Pseudogymnoascus destructans (Pd), that invades the skin of hibernating bats. While there are a number of treatments being researched, there is currently no effective treatment for WNS that is deployed in the field, except a few being tested on a limited scale. Bats have lowered immune function and response during hibernation, which may increase susceptibility to infection during the winter months. Antimicrobial peptides (AMPs) are a crucial component of the innate immune system and serve as barriers against infection. AMPs are constitutively expressed on skin and facilitate wound healing, stimulate other immune responses, and may also stay active on bat skin during hibernation. AMPs are expressed by all tissues, have direct killing abilities against microbes, and are a potential treatment for bats infected with Pd. In this investigation, the fungicidal activity of several readily available commercial AMPs were compared, and killing assay protocols previously investigated by Frasier and Lake were replicated to establish a control trial for use in future killing assays. Another aim of this investigation was to synthesize a bat-derived AMP for use in the killing assay. Sequences of bat-derived AMPs have been identified in bat skin samples obtained from a large geographic sampling of susceptible and resistant species. Contact was made with GenScript Inc., the company from which commercially available AMPs were purchased, to determine the characteristics of peptide sequences needed to synthesize an AMP for lab use. Based on recommendations from GenScript Inc., peptide sequences need to have a hydrophobicity of less than 50% and a sequence length of less than 50 amino acids. These criteria serve as a potential barrier because none of the known bat-derived sequences analyzed satisfy both of these requirements. The final aim of this study was to generate a conceptual model of the immune response molecules activated when bats are exposed to a fungal pathogen such as Pd. Overall, this work investigated sources of variability between trials of the killing assay, analyzed known bat-derived peptide sequences, and generated a conceptual model that will serve as a guideline for identification of immune response molecules on the skin of bats in future proteomics work.

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2019-05

Tell It to the Frogs: Fukushima’s nuclear disaster and its impact on the Japanese Tree Frog

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“Tell It to the Frogs: Fukushima’s nuclear disaster and its impact on the Japanese Tree Frog” is a representation of the work from Giraudeau et. al’s “Carotenoid distribution in wild Japanese tree frogs (Hyla japonica) exposed to ionizing radiation in

“Tell It to the Frogs: Fukushima’s nuclear disaster and its impact on the Japanese Tree Frog” is a representation of the work from Giraudeau et. al’s “Carotenoid distribution in wild Japanese tree frogs (Hyla japonica) exposed to ionizing radiation in Fukushima.” This paper looked to see if carotenoid levels in the tree frog’s vocal sac, liver, and blood were affected by radiation from Fukushima’s power plant explosion. Without carotenoids, the pigment that gives the frogs their orange color on their necks, their courtship practices would be impacted and would not be as able to show off their fitness to potential mates. The artwork inspired by this research displayed the tree frog’s degradation over time due to radiation, starting with normal life and ending with their death and open on the table. The sculptures also pinpoint where the carotenoids were being measured with a brilliant orange glaze. Through ceramic hand building, the artist created larger than life frogs in hopes to elicit curiosity about them and their plight. While the paper did not conclude any changes in the frog’s physiology after 18 months of exposure, there are still questions that are left unanswered. Why did these frogs not have any reaction? Could there be any effects after more time has passed? Is radiation leakage as big of a problem as previously thought? The only way to get the answers to these questions is to be aware of these amphibians, the circumstances that led them to be involved, and continued research on them and radiation.

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2019-05

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Understanding Differences Between Susceptibility and Resistance to White-Nose Syndrome in Bats: Methodological Optimization

Description

White-nose syndrome (WNS) is a fungal disease that infects hibernating bats of multiple species across large portions of eastern North America. To date, WNS has been responsible for the deaths of over seven million bats. It is not yet known

White-nose syndrome (WNS) is a fungal disease that infects hibernating bats of multiple species across large portions of eastern North America. To date, WNS has been responsible for the deaths of over seven million bats. It is not yet known why certain species are able to resist infection. Since the fungus invades the skin and some resistant species show no signs of the characteristic cutaneous lesions, it seems likely that resistant species contain specific defense mechanisms within their skin, such as antimicrobial peptides (AMPs) and other immunologically relevant proteins expressed by specific cell types or as secreted soluble components. Proteomics could be a useful tool for understanding differences in susceptibility, and could help identify AMPs that could be synthesized and used as control agents against the spread of the causative fungus. This study is the first to optimize proteomics methods for bat wing tissues in order to compare the skin proteomes of species variably impacted by WNS, including those of two endangered species. Further tests are planned to investigate methods of increasing protein yield without altering the size of the tissue sample collected, as well as the analysis of mass spectrometry data from processed skin tissues of five bat species differentially affected by WNS.

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2018-05

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Investigating Antimicrobial Controls for Bat White-Nose Syndrome

Description

Across large areas of eastern and midwestern North America, a severe reduction in multiple populations of bat species has been observed as the result of the emerging fungal disease, white-nose syndrome (WNS). WNS is caused by a psychrophilic (i.e. cold

Across large areas of eastern and midwestern North America, a severe reduction in multiple populations of bat species has been observed as the result of the emerging fungal disease, white-nose syndrome (WNS). WNS is caused by a psychrophilic (i.e. cold loving) fungus, Pseudogymnoascus destructans (Pd), that invades the skin of bats during hibernation. Recent studies have shown that during hibernation, bats have decreased immune system activity which would suggest increased susceptibility to infection. Antimicrobial peptides (AMPs) are an important component of the innate immune system and are expressed constitutively within all tissues that serve as barriers against infection. Killing pathogens at the level of the skin could prevent the need for more complex immune responses likely inhibited during hibernation, and therefore AMPs could be critical in combating infection by Pd and reducing population loss of susceptible bat species. In this investigation, the fungicidal activity of commercially available AMPs derived from the skin of multiple taxa, including amphibians, catfish, and humans were compared in order to study immunity at the level of the skin. Additionally, our aim was to create optimal methods for a low-cost antimicrobial-assay protocol that would provide quantitative results. We found that killing abilities at various concentrations of dermaseptin S-1 against Ca ATCC 10231 were consistent with literature values, while our values for magainin 2 and parasin 1 were far from the values previously recorded by other studies. It is possible that some differences can be accounted for by the difference in antimicrobial assay procedures, but our findings suggest potential differences to the well-known killing abilities of certain peptides nonetheless. Overall, the protocol established for the antimicrobial assays using serial dilutions and Sabouraud Dextrose plates was successful.

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2018-05

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Investigating Antimicrobial Controls for Bat White-Nose Syndrome

Description

Across large areas of eastern and midwestern North America, a severe reduction in multiple populations of bat species has been observed as the result of the emerging fungal disease, white-nose syndrome (WNS). WNS is caused by a psychrophilic (i.e. cold

Across large areas of eastern and midwestern North America, a severe reduction in multiple populations of bat species has been observed as the result of the emerging fungal disease, white-nose syndrome (WNS). WNS is caused by a psychrophilic (i.e. cold loving) fungus, Pseudogymnoascus destructans (Pd), that invades the skin of bats during hibernation. Recent studies have shown that during hibernation, bats have decreased immune system activity which would suggest increased susceptibility to infection. Antimicrobial peptides (AMPs) are an important component of the innate immune system and are expressed constitutively within all tissues that serve as barriers against infection. Killing pathogens at the level of the skin could prevent the need for more complex immune responses likely inhibited during hibernation, and therefore AMPs could be critical in combating infection by Pd and reducing population loss of susceptible bat species. In this investigation, the fungicidal activity of commercially available AMPs derived from the skin of multiple taxa, including amphibians, catfish, and humans were compared in order to study immunity at the level of the skin. Additionally, our aim was to create optimal methods for a low-cost antimicrobial-assay protocol that would provide quantitative results. We found that killing abilities at various concentrations of dermaseptin S-1 against Ca ATCC 10231 were consistent with literature values, while our values for magainin 2 and parasin 1 were far from the values previously recorded by other studies. It is possible that some differences can be accounted for by the difference in antimicrobial assay procedures, but our findings suggest potential differences to the well-known killing abilities of certain peptides nonetheless. Overall, the protocol established for the antimicrobial assays using serial dilutions and Sabouraud Dextrose plates was successful.

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Date Created
2018-05

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Investigating the Skin Immune Proteome of the White-Nose Syndrome Resistant Gray Bat, Myotis grisescens

Description

White-nose syndrome (WNS) is a cutaneous fungal infection caused by Pseudogymnoascus destructans (Pd) which was first observed in the United States in 2006. Pd infects bats during hibernation and leads to the development of cutaneous lesions and behavioral changes that

White-nose syndrome (WNS) is a cutaneous fungal infection caused by Pseudogymnoascus destructans (Pd) which was first observed in the United States in 2006. Pd infects bats during hibernation and leads to the development of cutaneous lesions and behavioral changes that can result in the animal's death. This study generated the first complete bat skin proteome for the WNS resistant gray bat (Myotis grisescens) to optimize sample preparation methods and identify immune proteins that may signal resistance. Wing tissue was collected from a female gray bat and processed in a Barocycler using 4M or 8M urea followed by an in-gel trypsin digestion of pooled samples and processing of separate samples without digestion specifically to capture and identify small antimicrobial peptides. Both undigested and digested samples were analyzed using a Thermo Fisher LTQ Orbitrap Velos mass spectrometer and interpreted using PEAKS software. A total of 29 immune proteins were identified including the antimicrobial peptide dermcidin. This method will be applied to a larger range of samples from five species variably impacted by WNS to compare skin proteomes with the aim of identifying immune proteins that are responsible for resistance at the barrier where Pd invades.

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2017-05

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Expressing Equilibrium

Description

The study of broad therapeutic advantages of dance is a growing field of interdisciplinary study. Yet, direct health benefits of dance from a molecular standpoint are still largely unknown. Literature review of dance performance displays in birds as well as

The study of broad therapeutic advantages of dance is a growing field of interdisciplinary study. Yet, direct health benefits of dance from a molecular standpoint are still largely unknown. Literature review of dance performance displays in birds as well as other creatures and use of creative tools to analyze the diverse, lifelong experiences of dancers helped shed some light on the subject. Although dance experience exposes harms tied to the social constraints of how the form is experiences buried under joyful takeaways of dance, research supports overall health benefits from moderate amounts of dance maintained in perfect equilibrium.

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2022-05

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Quantifying the Preference of Corynorhinus townsendii to Hibernate in Highly Ventilated Areas in Arizona Caves

Description

Corynorhinus townsendii, a bat species residing in north-central Arizona, has historically been observed hibernating in highly ventilated areas within caves and abandoned mines, but there is little to no specific data regarding this tendency. Understanding how air movement may influence

Corynorhinus townsendii, a bat species residing in north-central Arizona, has historically been observed hibernating in highly ventilated areas within caves and abandoned mines, but there is little to no specific data regarding this tendency. Understanding how air movement may influence hibernacula selection is critical in bettering conservation efforts for Arizona bats, especially with white-nose syndrome continuing to devastate bat species populations throughout the United States. My study aimed to begin filling in this knowledge gap. I measured wind speed in three known Arizona hibernacula during the winter hibernation season and combined this data with the locations of bats observed throughout each of the three survey locations. I modeled our findings using a generalized linear model, which confirmed that wind speed is indeed a predictor of C. townsendii roost selection.

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2022-05