A broad-spectrum mer operon in a multi-drug resistant strain of the fish pathogen, Aeromonas salmonicida

Aeromonas salmonicida AS03, a potential fish pathogen, was isolated from Atlantic salmon, Salmo salar, in 2003. This strain was found to be resistant to ≥1000 mM HgCl2 and ≥32 mM phenylmercuric acetate as well as multiple antimicrobials. Mercury (Hg) and antibiotic resistance genes are often located on the same mobile genetic elements, so the genetic determinants of both resistances and the possibility of horizontal gene transfer were examined. Specific PCR primers were used to amplify and sequence distinctive regions of the mer operon. A. salmonicida AS03 was found to have a pDU1358-like broad-spectrum mer operon, containing merB as well as merA, merD, merP, merR and merT, most similar to Klebsiella pneumonaie plasmid pRMH760. To our knowledge, the mer operon has never before been documented in Aeromonas spp. PCR and gene sequencing were used to identify class 1 integron associated antibiotic resistance determinants and the Tet A tetracycline resistance gene. The transposase and resolvase genes of Tn1696 were identified through PCR and sequencing with Tn21 specific PCR primers. We provide phenotypic and genotypic evidence that the mer operon, the aforementioned antibiotic resistances, and the Tn1696 transposition module are located on a single plasmid or conjugative transposon that can be transferred to E. coli DH5α through conjugation in the presence of low level Hg and absence of any antibiotic selective pressure. Additionally, the presence of low-level Hg or chloramphenicol in the mating media was found to stimulate conjugation, significantly increasing the transfer frequency of conjugation above the transfer frequency measured with mating media lacking both antibiotics and Hg. This research demonstrates that mercury indirectly selects for the dissemination of the antibiotic resistance genes of A. salmonicida AS03.

Photochemical Generation and Intramolecular Chemistry of β-Alkoxycarbenes

The photolytic phenanthrene-based precursors for both β-methoxycarbene and β-ethoxycarbene were synthesized with and without a deuterium label attached to the a carbon. The incorporation of this deuterium label allowed distinction between a 1, 2-H shift and a 1, 2-O shift pathway to the respective alkyl vinyl ether, without the influence of a primary kinetic isotope effect. Photolyses of these precursors gave rearrangement products of the expected β-alkoxycarbenes. In the case of β-methoxycarbene, no methyl vinyl ether was observed due to its volatility. However, the appearance of aldehyde peaks in the NMR spectra, from an apparent further rearrangement to acetaldehyde through an enol intermediate, indicated that a 1,2-H shift had occurred. Ethyl vinyl ether was isolated following the photolysis of the β-ethoxycarbene precursor. Quantification of the two pathways showed less than 2% undergoing an ethoxy shift to the ethyl vinyl ether. Yield experiments on this photolysis demonstrated a maximum yield of β-ethoxycarbene as 43%, though this decreased as the experiment continued. Computational work on the β-ethoxycarbene system indicates that the triplet scate is more stable than the singlet. In addition, the activation energy to the 1.2-H shift pathway is remarkably low and is clearly consistent with the observed overwhelming preference for this pathway in the experiment.

Salmoninae Commensal Bacteria Show a Positive Correlation Between Antibiotic Resistance and Mercury Resistance Profiles

Bacterial isolates from natural sites with high toxic and heavy metal contamination more frequently contain determinants for resistance to antimicrobials. Natural strains were isolated from the ingesta and external slime of Salmo salar (Linnaeus, 1758) and Salvelinusjontinalis (Mitchell, 1814). Fish specimens were acquired from Casco Bay hatcheries, Casco, ME where there is no history of antibiotic use. Seventy-nine bacterial strains, including many well-documented salmonid commensals (an association from which the fish derives no benefit), were identified using 165 rRNA gene sequencing. Mercury resistant isolates were selected for initially on 25μM HgCI2. Strains were then grown at 20-24°C on Trypticase Soy Agar (TSA) plates containing 0-1000μM HgCl2 or 0-130μM Phenyl Mercuric Acetate (PMA). Mercury in the hatchery feed water due to ubiquitous non-point source deposition has selected for the mercury resistance observed in bacterial strains. Antibiotic resistance determinations, as measured by Minimum Inhibitory Concentration MIC) assays were performed on the 79 bacterial isolates using Sensititrel antimicrobial susceptibility panels. A positive linear correlation between the mercury (pMA and HgCl2) MIC's and antibiotic resistance for all observed strains was demonstrated. Conjugation experiments with Pseudomonas, Aeromonas, and Azomonas donors confirmed phenotypic transfer of penicillin and cephem resistances to Escherichia coli DH5a recipients. Conjugation experiments with Pseudomonas donors showed minimal transfer of tetracycline and minoglycoside resistances to Escherichia coli DH5a recipients. Our study suggests that the accumulation of antimicrobial resistances observed in these natural bacterial populations may be due to the indirect selective pressure exerted by environmental mercury.

Pressure Perturbation Calorimetry and Guest-Host Chemistry of NDMG and N-MAP with p-Sulfonatocalix[6]arene

The aim of this project is to provide an explanation for recently obtained binding constants for two similar guest molecules, NDMG and N-MAP, with a p-sulfonatocalix[6]arene host in ammonium acetate buffer. This work was done primarily using pressure perturbation calorimetry, which is a technique that determines the coefficient of thermal expansion, α, which is in turn related to the solute molecule's effect on the order of the surrounding water molecules. A series of experiments were designed to test the effects of suspected confounding variables on the validity of PPC data. PPC was then used to study NDMG and N-MAP in ammonium acetate buffer. NDMG exhibited a minimum in α as function of temperature, while N-MAP did not. This difference was theorized to be due to the formation of an intramolecular hydrogen bond in monocationic NDMG that would lower the heat capacity of the molecule and better distribute the molecule's charge. Computational work and nuclear magnetic resonance spectroscopy confirmed that monocationic, ring-closed NDMG has less concentrated charge and more constrained motion than monocationic, ring-open NDMG. This evidence supports the theory that monocationic NDMG forms an intramolecular hydrogen bond and that this may be responsible for the minimum in α. This difference may explain the differences in binding constants between NDMG and N-MAP.

Estimating Stormwater Runoff for Community Gardens in New York City

While much of the literature cites community gardens as providing urban ecosystem services, there is very little research quantifying these benefits. This thesis compares the stormwater runoff rates of urban vacant lots, community gardens, and residential developments in New York City and evaluates community gardens as green infrastructure.

Using Spatiotemporal Methods to Fill Gaps In Energy Usage Interval Data

Researchers analyzing spatiotemporal or panel data, which varies both in location and over time, often find that their data has holes or gaps. This thesis explores alternative methods for filling those gaps and also suggests a set of techniques for evaluating those gap-filling methods to determine which works best.

Utilizing Remote Sensing Imagery to Monitor Vegetation Change within World Heritage Sites

ABSTRACT World Heritage sites provide a glimpse into the stories and civilizations of the past. There are currently 1007 unique World Heritage properties with 779 being classified as cultural sites, 197 as natural sites, and 31 falling into the categories of both cultural and natural sites (UNESCO & World Heritage Centre, 1992-2015). However, of these 1007 World Heritage sites, at least 46 are categorized as in danger and this number continues to grow. These unique and irreplaceable sites are exceptional because of their universality. Consequently, since World Heritage sites belong to all the people of the world and provide inspiration and admiration to all who visit them, it is our responsibility to help preserve these sites. The key form of preservation involves the individual monitoring of each site over time. While traditional methods are still extremely valuable, more recent advances in the field of geographic and spatial technologies including geographic information systems (GIS), laser scanning, and remote sensing, are becoming more beneficial for the monitoring and overall safeguarding of World Heritage sites. Through the employment and analysis of more accurately detailed spatial data, World Heritage sites can be better managed. There is a strong urgency to protect these sites. The purpose of this thesis is to describe the importance of taking care of World Heritage sites and to depict a way in which spatial technologies can be used to monitor and in effect preserve World Heritage sites through the utilization of remote sensing imagery. The research conducted in this thesis centers on the Everglades National Park, a World Heritage site that is continually affected by changes in vegetation. Data used include Landsat satellite imagery that dates from 2001-2003, the Everglades' boundaries shapefile, and Google Earth imagery. In order to conduct the in-depth analysis of vegetation change within the selected World Heritage site, three main techniques were performed to study changes found within the imagery. These techniques consist of conducting supervised classification for each image, incorporating a vegetation index known as Normalized Vegetation Index (NDVI), and utilizing the change detection tool available in the Environment for Visualizing Images (ENVI) software. With the research and analysis conducted throughout this thesis, it has been shown that within the three year time span (2001-2003), there has been an overall increase in both areas of barren soil (5.760%) and areas of vegetation (1.263%) with a decrease in the percentage of areas classified as sparsely vegetated (-6.987%). These results were gathered through the use of the maximum likelihood classification process available in the ENVI software. The results produced by the change detection tool which further analyzed vegetation change correlate with the results produced by the classification method. As well, by utilizing the NDVI method, one is able to locate changes by selecting a specific area and comparing the vegetation index generated for each date. It has been found that through the utilization of remote sensing technology, it is possible to monitor and observe changes featured within a World Heritage site. Remote sensing is an extraordinary tool that can and should be used by all site managers and organizations whose goal it is to preserve and protect World Heritage sites. Remote sensing can be used to not only observe changes over time, but it can also be used to pinpoint threats within a World Heritage site. World Heritage sites are irreplaceable sources of beauty, culture, and inspiration. It is our responsibility, as citizens of this world, to guard these treasures.

Comparative Analysis of Adolescent Pregnancy in Central America, Mexico and South Carolina

This paper addresses the issue of adolescent pregnancy in Mexico, Central America and South Carolina and implications for social work practice with immigrant communities. The paper is based on current literature and on cross-national, on-line survey of local and international pregnancy prevention programs. The paper analyzes and discusses various psychosocial causes of pregnancy in adolescents, including: limited opportunities for formal education, infrequent open discussions about sexual health, rising costs of adequate birth control, and difficulty in obtaining contraceptives in remote locations. This research paper analyzes current statistics on the effectiveness of existing projects and programs and compares and contrasts research about the validity and efficacy of these programs in both South Carolina and abroad. Finally, the paper addresses implications for social work practice with adolescents in immigrant communities.

Undergraduate Scholarship at Winthrop University 2012 Book of Abstracts

Winthrop University proudly presents the first Undergraduate Scholarship at Winthrop University Book of Abstracts. Building off the nine-year tradition of producing abstract books for students in the College of Arts and Sciences, University College is now creating this book to present the scholarship occurring throughout all five academic colleges in the university: College of Arts and Sciences (CAS), College of Business Administration (CBA), College of Education (COE), the College of Visual and Performing Arts (CVPA) and University College (UC). In addition to the research abstracts, we are using the book to document the students who have completed Honors Theses, applied for Nationally Competitive Awards, and were selected as McNair Scholars.