{"title":"Conservation and Taxonomic Assessment of an Undescribed Crayfish in Coastal Virginia","authors":"Alyssa Oppedisano","doi":"10.55632/pwvas.v94i1.930","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.930","url":null,"abstract":"Creaserinus fodiens (Cottle 1863), commonly known as the digger crayfish, is a primary burrower that inhabits complex burrows in wetlands, seasonal pools, wooded floodplains, and roadside ditches. Historically, C. fodiens have been found from Ontario, Canada following the United States down to Texas and across four Atlantic slope states including Virginia, Maryland, North Carolina, and South Carolina. The Atlantic slope clades are geographically isolated from other C. fodiens populations by the continental shelf on the East coast and by the Blue Ridge Mountains. Despite having a wide geographical range, little research has been done on the Atlantic slope clades of Creaserinus spp. Recent genetic and morphological data suggests that the Virginia population of C. fodiens could be described as a different species. Additional morphological and genetic data from gill samples will lead to a better understanding of where the separation in classification lies among the Virginia population. A conservation and taxonomic assessment for Creaserinus fodiens along the Virginia Atlantic slope will be constructed from the results. These assessments will contribute to our understanding of global crayfish biodiversity and provide important biological insight to the management and conservation of these species.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"163 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86794850","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Elio Delatore III, E. Roberts, Joseph Horzempa, S. Cantlay
{"title":"Deletion of FTL_1199 to determine the role of this gene in erythrocyte invasion by Francisella tularensis.","authors":"Elio Delatore III, E. Roberts, Joseph Horzempa, S. Cantlay","doi":"10.55632/pwvas.v94i1.872","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.872","url":null,"abstract":"Francisella tularensis is a bacterium that induces the zoonotic disease tularemia. In the course of infection, F. tularensis bacteria invade erythrocytes, a phenomenon that heightens the colonization of ticks after a blood meal. To better understand the mechanism of erythrocyte invasion, we hypothesized that transcription of bacterial genes significant in erythrocyte invasion would be upregulated upon exposure to these host cells. An RNA-seq unveiled that transcription of 7% of F. tularensis genes augment when in erythrocyte presence. Of these, we pinpointed three putative transcriptional regulators, namely FTL_0671, FTL_1199, and FTL_1665. The goal was to delete FTL_1199 in F. tularensis LVS. Splicing by overlap extension PCR amplified and duplicated the up and downstream (~500 bp each) regions of the target gene in tandem into a shuttle vector that is insecure within F. tularensis. This newly generated plasmid, pDEL1199, was mobilized inside of F. tularensis by conjugation. Merodiploid strains generated by homologous recombination were isolated and transformed with pGUTS – a stable plasmid that encodes a homing endonuclease (I-SceI) and a kanamycin resistance cassette. Expression of I-SceI within the merodiploid produces a double-stranded break in pDEL1199 that had previously integrated in the chromosome. This breakage resulted in a second recombination that either ensued to wild-type or deletion of FTL_1199 deduced through a PCR. Finally, in DFTL_1199 strains, pGUTS was cured by successive cultivation in the absence of selection followed by replica-plating on chocolate II agar ± kanamycin. Gentamicin protection assays involving F. tularensis DFTL_1199 suggest that FTL_1199 is important in erythrocyte invasion. \u0000 \u0000(Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence, R15HL14735 from NHLBI, and funds from the NASA West Virginia Space Grant Consortium).","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"90 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76191085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
N. Storer, Justin Sanclemente, B. Ringhiser, Zachary J. Loughman
{"title":"The Microhabitat Preferences Based on Relative Density and Stream Assessment of Regina septemvittata in a small West Virginia Stream","authors":"N. Storer, Justin Sanclemente, B. Ringhiser, Zachary J. Loughman","doi":"10.55632/pwvas.v94i1.913","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.913","url":null,"abstract":"Regina septemvittata is a slim-bodied water snake in the family Natricidae that is native to the northeastern United States. Being the only species to consume exclusively freshly molted crayfish, R. septemvittata is categorized as a specialist. The selectivity of prey suggests possible other narrow range preferences including refuge and hunting locations. This study is being conducted in the North Fork of Short Creek in Ohio County, WV. In addition, Nerodia sipedon sipedon is also found within the study stream system. N. s. Sipedon is a generalist water snake species that has been shown to be larger and more aggressive than R. septemvittata. Both species are captured throughout their activity period, April - November, starting in 2018. In addition to environmental and morphological data being recorded at the time of collection, qualitative habitat evaluation indexes (QHEI) were recorded at the initiation of the project. QHEI allows for a more standardized evaluation of the quality of the stream system. The combination of the quality of the stream in 100-meter sections of the stream in addition to snake location data can show relative preferences of R. septemvittata throughout the stream system. The current data suggests a greater preference for disturbed habitat as well as a higher frequency of hot spots in the stream as opposed to their generalist counterparts N. s. sipedon. ","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"33 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88798827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The Effect of Efflux Pumps and Oxidative Stress on Neisseria gonorrhoeae Susceptibility to Resazomycins","authors":"J. Gibson, E. Young, Deanna M Schmitt, J. Rice","doi":"10.55632/pwvas.v94i1.873","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.873","url":null,"abstract":"The increasing prevalence of multidrug-resistant Neisseria gonorrhoeae strains underscores the need for novel antimicrobials against this pathogen. Resazomycins, derivatives of resazurin (Rz), have shown robust antimicrobial activity against N. gonorrhoeae (Ng) in vitro. In vivo, however, resazomycins exhibit limited efficacy in a mouse model of gonorrhea. Previous experiments have shown that N. gonorrhoeae is more resistant to resazurin at oxygen levels comparable to those seen in host tissue (2%). We hypothesized this difference in susceptibility at low oxygen compared to atmospheric oxygen (~20%) was due to altered activity of multi-drug efflux pumps. To test this, we screened a selection of N. gonorrhoeae mutants that do not express or overexpress either the MtrCDE or NorM efflux pumps for Rz susceptibility. Overexpression of MtrCDE resulted in increased resistance to Rz at both 2% and ~20% oxygen suggesting resazurin may be a substrate of this efflux pump. Loss of expression of either MtrCDE or NorM had no effect on the increased resistance of N. gonorrhoeae to resazurin at low oxygen. We next sought to determine whether the increased susceptibility of Rz at 20% oxygen is due to oxidative stress. To test this, we measured the susceptibility of N. gonorrhoeae to Rz in the presence and absence of the antioxidants, cysteine HCl and glutathione, at 20% oxygen. In the presence of cysteine HCl or glutathione, multiple N. gonorrhoeae strains had a higher Rz MIC at 20% oxygen. Here, we have shown oxygen concentration affects N. gonorrhoeae susceptibility to Rz due to increased oxidative stress. ","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"34 2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91197772","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Claire O. Kelly, E. Young, J. Gibson, S. McGovern, Emma Beatty, Kendall Soulder, J. Rice, Ryan J. Percifield, D. Primerano, Nicole L. Garrison, Deanna M Schmitt
{"title":"Role of FTL_0895 in Francisella tularensis Susceptibility to Resazurin.","authors":"Claire O. Kelly, E. Young, J. Gibson, S. McGovern, Emma Beatty, Kendall Soulder, J. Rice, Ryan J. Percifield, D. Primerano, Nicole L. Garrison, Deanna M Schmitt","doi":"10.55632/pwvas.v94i1.892","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.892","url":null,"abstract":"Resistance to antibiotic treatments coupled with the decline in antibiotic discovery has resulted in a steady increase in deaths caused by once “curable” bacterial infections. Developing new drugs is crucial to prevent more loss of life in the future. We discovered the compound resazurin exhibits antimicrobial activity against gram-negative bacteria including Francisella tularensis (Ft), however, certain strains of Ft have developed resistance to resazurin. Understanding how Ft develops resistance to resazurin will help with defining the mechanism by which resazurin elicits its antimicrobial effect. Whole genome sequencing of resazurin-resistant (Rzr) Ft LVS mutants revealed four mutations found in 93% of the isolates sequenced. Three mutations were within the coding regions of FTL_0421, FTL_0895, and FTL_1504 and the other mutation was 50 bp upstream of FTL_0445, likely disrupting expression of this gene. The focus of my project was to explore the role of FTL_0895 in resazurin susceptibility. To confirm this gene plays a role in the reduced susceptibility of the Rzr strains to resazurin, we cloned the wild-type copy of FTL_0895 into the Francisella vector pABST which contains the robust groE promotor of Ft. The resulting plasmid will be electroporated into one of the Rzr mutants and we will test the susceptibility of the complemented strain to resazurin, using time kill and agar dilution assays. If the susceptibility of the complemented strain to resazurin is restored, then it can be determined that FTL_0895 is a potential target of resazurin.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"16 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88026451","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Spatial dynamics of an invasive earthworm (Amynthas sp.) in West Liberty University’s campus woods community.","authors":"Jason Ake, L. Butcher, J. Dann","doi":"10.55632/pwvas.v94i1.880","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.880","url":null,"abstract":"Asian pheretimoid earthworms (Amynthas sp.) have been reported in the hardwood forests of the eastern United States with increasing frequency. These invasions can have highly negative ecosystem-wide impacts. Amynthas sp. are prolific breeders capable of reaching high densities in forest soils. They consume large quantities of leaf litter, disturbing natural soil horizons, draining available nutrients and damaging plant communities. In the summer and fall of 2021, population surveys were conducted in an area of West Liberty University’s campus woods community known to be infested with pheretimoid worms. Spatial dynamics were examined using SADIE analysis. It was found that Amynthas sp. showed a trend towards aggregation in both the summer and fall, with a more evident trend towards aggregation in the fall. An analysis of spatial association showed that the distributions between consecutive samples were positively associated. This indicates stability of aggregations over short timescales in both summer and fall. Understanding the population dynamics of Amynthas sp. and their interactions with native communities will aid in the development of management strategies to prevent the spread of this invasive and reduce the disturbance of native forest ecosystems.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"46 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83601614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Synchronizing the Teaching Resources of Energy Conservation Principle in Mechanical Engineering Courses","authors":"Y. Panta, W. Church","doi":"10.55632/pwvas.v94i1.896","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.896","url":null,"abstract":"The conservation of energy, the conservation of mass, and the conservation of momentum are three fundamental concepts (or laws) of physics that are regularly reviewed in several undergraduate engineering courses. Mechanical energies in the form of kinetic and potential forms are the most easily understood forms of energy in engineering dynamics courses. Fluid flow energies related to pressure, velocity, elevation, fluid friction, pump, and turbine are covered in a fluid mechanics course. In a thermodynamics course, the first law deals with heat transfer and work done that causes a change of internal energy in a system. Aerospace engineers normally simplify a thermodynamic analysis by using intensive variables also called specific variables. In all these courses, the conservation of energy states that the amount of energy remains constant, that means that energy is neither created nor destroyed but transferable from one form to another, keeping the total energy same within a fixed domain. \u0000In several instances, students are either misunderstood or unclear about energy and its conservation concepts, however those very concepts are reviewed over and over in multiple courses. Through an integrative teaching approach that maps the smooth flow of energy and its conservation concepts in several undergraduate mechanical engineering courses, we are relating our shared teaching resources of the energy conservation principle. In this session, we present our pilot study on the synchronization of resources teaching the energy conservation principle in a sequence of undergraduate courses and our mitigation plan to clear up students’ misunderstanding on the energy conservation.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"13 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90049091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Thyroxine Metabolism in an Avian Model of Induced Thyrotoxicosis","authors":"Kirsten Bogunovich, Holly L. Racine, Peri Gray","doi":"10.55632/pwvas.v94i1.875","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.875","url":null,"abstract":"Craniosynostosis (CS) is the premature fusion of cranial sutures which leads to abnormal cranial structure and causes abnormal neurocognition and behavior. Maternal thyroid disorders are associated with incidence of CS in infants. However, the mechanism of TH-induced CS is still obscure. Thyroxine (T4) metabolism is regulated by the expression of deiodinases. Type I and type II deiodinases (DIO1 and DIO2) remove iodine from T4 producing active triiodothyronine (T3). Deiodinase III (DIO3) inactivates both T4 and T3. Our lab is establishing a model to study the effects of thyrotoxicosis induced CS using fertilized chicken eggs. Eggs were injected into the air cell with either saline or 25 ng T4 on days E11 and E15 of embryonic development. Heart rates were measured daily from EKG recordings and on E19, tissue samples were collected for qRT-PCR analysis. The main objective was to validate the induced state of thyrotoxicosis by evaluating negative feedback mechanisms of thyroid hormone regulation following exposure by measuring expression of DIO1 and DIO3 in embryonic livers. Since livers are the main site of metabolism, we hypothesize that levels of DIO3 will increase following thyroxine exposure. Results demonstrated a significant upregulation of DIO3 2 days after exposure and a significant downregulation 4 days following exposure. No significant differences were observed in DIO1 on any of the days after exposure. These results, along with correlating heart rates, support our model by of induced thyrotoxicosis by demonstrating the metabolism of thyroxine in response to our treatment regimen. \u0000 \u0000 \u0000Acknowledgment of NASA West Virginia Space Grant Consortium (Grant #80NSSC20M0055) and the Genomics Core Facility and WV-INBRE (NIH grant P20GM103434). \u0000","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"66 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91232784","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Behavioral ecology of Ixodes scapularis and the implications for Lyme disease","authors":"My-Van Tran, Kim Bjorgo-Thorne, M. Sal","doi":"10.55632/pwvas.v94i1.934","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.934","url":null,"abstract":"The most common ticks in West Virginia include the American dog tick (Dermacentor variabilis), the blacklegged tick (Ixodes scapularis), and the brown dog tick (Rhipicephalus sanguineus ( (WVU extension). One of the most common tick-borne diseases in WV is Lyme disease, caused by the spirochete Borrelia burgdorferi. Ixodes scapularis is a carrier for the B burgdorferi pathogen. Therefore, species identification is vital. The purpose of this research is to understand how tick behavior can affect the transmission and distribution of Lyme disease in the eastern United States, specifically in West Virginia. An ethogram was constructed to display five core behaviors of Ixodes scapularis.Other behaviors may appear in the ethogram but those remain variable across species and were not included. No other behaviors distinctive to Ixodes scapularis were found. \u0000Five main behaviors were noted in Ixodes scapularis and can be functionally applied to other tick species. By understanding the behavior of ticks, the knowledge can be used to understand the prevalence of tick distribution in the eastern United States. When combining tick behavior and its distribution, one can then focus on the prevalence of Borrelia burgdorferi in infected ticks. Additionally, infected tick behavior can differ from uninfected ticks, resulting in more frequent questing and biting behaviors. Different populations of the same species can also differ in questing height and hiding behavior.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"29 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83725262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Creating a baseline for bioaccumulation in crayfish in the central Appalachian coalfields","authors":"T. Whitson","doi":"10.55632/pwvas.v94i1.921","DOIUrl":"https://doi.org/10.55632/pwvas.v94i1.921","url":null,"abstract":"Cambarus veteranus and Cambarus callainus are highly imperiled crayfish that are federally listed. These crayfish are native to the coalfields region of central Appalachia. The dominant industry and land use in this region is coal mining. The most common method of mining in this area, surface mining, employs large ponds to hold a slurry water byproduct created when cleaning the coal. Intermittently, the containment structures for these ponds fail causing the uncontrolled release of coal slurry. There is a lack of data on the rates of bioaccumulation of elements commonly associated with mining in crayfish within this region before and after these spills occur, such that there is no reference to know what concentrations constitutes “normal” rates. Chemical pollutants along with increased sedimentation from industrial sources are known to cause adverse effects in crustaceans and can be lethal. Therefore, it is crucial to better understand what baseline conditions of bioaccumulation are within this region before spills occur. This study assessed the bioaccumulation of magnesium (Mg), aluminum (Al), chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), zinc (Zn), arsenic (As), selenium (Se), cadmium (Cd), and lead (Pb) in non-imperiled crayfish by analyzing hepatopancreas and gill tissues. We analyzed rates of bioaccumulation grouped by crayfish genus and tissue type throughout each season Differences between genus and tissue type, reference and degraded sites, and between crayfish tissue type, periphyton, and water were also investigated. Seasonal variation in the concentrations of elements in crayfish tissue was found with higher element concentrations during winter and lower concentrations during summer. Differences between crayfish tissue type, periphyton, and water were also found, with periphyton typically having the highest concentrations, water having the lowest concentrations, and crayfish tissue being between concentrations found in water and periphyton.","PeriodicalId":92280,"journal":{"name":"Proceedings of the West Virginia Academy of Science","volume":"21 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83264007","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}