The question of post-mortem interval (PMI) or time since death is often the most sought after piece of information associated with a medical death investigation. Based on the observation that DNA degradation disproportionately affects the analysis of larger genetic loci, it was proposed that DNA degradation, as a result of autolysis or putrefaction, could prove suitable as a potential rate-of-change indicator of PMI. Nine randomly amplified polymorphic DNA (RAPD) analysis primers and three sets of directed amplification primers were evaluated to determine their suitability for use in assessing the degree of DNA fragmentation in tissue samples. They were assessed for amplicon specificity, total DNA target sensitivity, allele monomorphism and the observance of degradation-based profile changes. Markers meeting the requisite criteria were then used to assess a range samples degraded under controlled and uncontrolled conditions. Tissue samples collected from seven domestic pigs (Sus scrofa) were incubated under controlled laboratory or uncontrolled field conditions to produce samples simulating those potentially collected in a forensic case. DNA samples isolated from these specimens were then analyzed at those loci which had been determined to meet the requisite criteria. Collectively, data generated from these analyses indicate that genetic profiles generated by this approach can provide …

The objective of this research was to investigate lead speciation in the soil/sediment-water environment and to better understand how the species affect lead mobility under different environmental conditions. The research involved both field soil and sediment samples as well as standard lead solutions. Field samples were fully characterized and extracted by aqueous and organic solvents. The results were compared and evaluated with the metal speciation model, MINTEQA2. Hydrolytic polymerization and organic complexation studies were conducted with standard lead solutions under controlled experimental conditions. Results of the field samples showed that pH, dissolved cations, ionic strength, dissolved organic matter, and nature of the soil/sediment matrix play major roles in the distribution and mobility of lead (Pb) from contaminated sites. In the aqueous equilibration experiment, the magnitude of Pb2+ solubilization was in the order of pH4>pH7>pH9. The results were in good agreement with MINTEQA2 predictions. An important finding of the research is the detection of Pb polymerization species under controlled experimental conditions. At pH 5.22, Pb polymeric species were formed at rate of 0.03 per day. The role of Pb complexation with organic matter was evaluated in both field and standard samples. Different methodologies showed three types of organically bound Pb. A …

My study of the effects of carbaryl in aquatic systems under controlled laboratory conditions emphasized four major objectives: (1) to determine the acute toxicity of carbaryl to the herbivorous adult and immature Trichocorixa reticulata (Guerin)(Hemiptera: Corixidae), and to the omnivorous larvae of Glyptotendipes barbipes (Staeger) (Diptera: Chironomidae) under static bioassay; (2) to adapt a quantitative method of analysis for carbaryl in water and whole insect tissue extract; (3) to measure the accumulation of the insecticide by G. barbipes under static exposure; and (4) to quantify the uptake and loss of carbaryl by G. barbipes under daily-renewed sublethal dosages.

Hazard assessment of pesticides in aquatic environments requires accurate predictions of persistence and compartmentalization. A strategy for developing confidence in predictive fate models, such as the Exposure Analysis Modeling System (EXAMS) and the Simplified Lake and Stream Analysis model (SLSA), is to test the models using carefully chosen chemicals in semi-controlled and field situations. An objective of this approach would be to isolate the variability in a particular fate process and thereby assess the ability of an algorithm to model the process. For example, endothall, a relatively watersoluble aquatic herbicide, has essentially a sole fate process, biotransformation. Endothall was used to test the predictive capabilities of EXAMS and SLSA and to identify sources of variance in those predictions.

Despite the variety of successful reports of the preservation, recovery, and identification of archaeological proteins in general, there are few positive reports regarding mass spectrometry-based identification of ceramic-bound proteins. In large part, this shortage is due to the lack of consideration for the unique taphonomic histories of such residues and, in general, methods development. Further, because negative results are rarely published, there is no baseline to which results can be compared. This paper attempts to address these challenges via a multi-pronged approach that uses mass spectrometry and complementary approaches to evaluate ceramic-bound protein preservation in both controlled, actualistic experiments, and in archaeological artifacts. By comparing the results obtained from protein-spiked, experimentally-aged ceramic to those obtained from both faunal and ceramic archaeological materials, an enhanced perspective on protein preservation and subsequent recovery and identification is revealed. This perspective, focusing on taphonomy, reveals why negative results may be the norm for ceramic artifacts when non-targeted methods are employed, and provides insight into how further method development may improve the likelihood of obtaining positive results.

N-acyl ethanolamines (NAEs) are a class of lipids recently recognized as signaling molecules which are controlled, in part, by their degradation by fatty acid amide hydrolase (FAAH). On the basis of previous studies indicating increased NAE levels in a tobacco cell suspension-xylanase elicitor exposure system and the availability of FAAH mutants, overexpressor and knockout (OE and KO) genotypes in Arabidopsis thaliana, further roles of NAEs in A. thaliana plant defense was investigated. The commonly occurring urban antimicrobial contaminant triclosan (TCS) has been shown to suppress lipid signaling associated with plant defense responses. Thus, a second objective of this study was to determine if TCS exposure specifically interferes with NAE levels. No changes in steady state NAE profiles in A. thaliana-Pseudomonas syringae pv. syringae and A. thaliana-flagellin (bacterial peptide, flg22) challenge systems were seen despite evidence that defense responses were activated in these systems. There was a significant drop in enoyl-ACP reductase (ENR) enzyme activity, which catalyzes the last step in the fatty acid biosynthesis pathway in plants, on exposure of the seedlings to TCS at 10 ppm for 24 h and decreased reactive oxygen species (ROS) production due to flg22 in long term exposure of 0.1 ppm and short term …

Thauera aminoaromatica MZ1T, a floc-forming bacterium isolated from an industrial activated sludge wastewater treatment plant, overproduces exopolysaccharide (EPS) leading to viscous bulking. This phenomenon results in poor sludge settling and dewatering during the clarification process. To identify genes responsible for bacterial flocculation, a whole genome phenotypic sequencing technique was applied. Genomic DNA of MZ1T flocculation-deficient mutants were subjected to massively parallel sequencing. The resultant high-quality reads were assembled and compared to the reference genome of the wild type genome. We identified nine nonsynonymous mutations and one nonsense mutation putatively involved in EPS biosynthesis. Complementation of the nonsense mutation located in an EPS deacetylase gene restored the flocculating phenotype. The FTIR spectra of EPS isolated from the wild-type showed reduced C=O peak of the N-acetyl group at 1665 cm-1 as compared to the spectra of MZ1T floc-deficient mutant EPS, suggesting that the WT EPS was partially deacetylated. Gene expression analysis also demonstrated the deacetylase gene transcript increased before flocculation occurred. The results suggest that the deacetylation of MZ1T EPS is crucial for flocculation. The information obtained from this study will be useful for preventing viscous bulking and wastewater treatment system failure, and may have potential applications in the biotechnology sector for …

The physiological characteristics and organization of cat primary somatosensory cortex (SI) were studied in electrophysiological and anatomical experiments. In single cell recording experiments, quantitatively controlled mechanical stimuli were used to examine the responses of SI cortical neurons to the velocity component of skin or hair displacement. The firing frequency of most rapidly adapting neurons increased as stimulus velocity was increased. Rapidly adapting neurons were classified based on their response patterns to constant-velocity ramp stimuli. Neurons in these classes differed significantly in sensitivity to stimulus velocity and amplitude, adaptation rate, and spontaneous firing rate. The results suggest that frequency coding of stimulus displacement velocity could be performed by individual SI rapidly adapting neurons, and that the classes of rapidly adapting neurons may play different roles in sensation of tactile stimuli. Tract-tracing experiments were used to investigate the ipsilateral corticocortical connections of areas 3b and 2 in SI. Different patterns of connections were found for these areas: area 2 projects to areas 3b, 1, 3a, 5a, 4 and second somatosensory cortex (SII), and area 3b projects to areas 2, 1, 3a and SII. To further compare the organization of these areas, the thalamic input to the forepaw representation within each area was …

Methods to determine genetic diversity and relatedness within populations are essential tools for proper wildlife management. Today the approach of choice is polymerase chain reaction-based microsatellite analysis. Seven new polymorphic loci were isolated from a microsatellite-enriched Caribbean flamingo genomic library and used to characterize survey populations of Caribbean and African greater flamingos. In addition, four of these loci were used to verify parentage relationships within a captive-breeding population of African greater flamingos. Parentage predictions based upon gamekeeper observations of breeding and nesting did not always agree with genetic-based parentage analyses of the nine suggested family groups. Four family groups were supported (groups I, II, III and VI) by there results. However, an analysis of the remaining five suggested groups, with a total of eight offspring/dam and eight offspring/sire suggested relationships, yielded seven exclusions of the suggested dam and six exclusions of the suggested sire. This put the overall suggested dam exclusion rate at 35% and exclusion rate for suggested sires at 29%. Although the keeper observation data for our family groups must be considered a variable of concern at this time, these findings are certainly suggestive that more carefully controlled studies may reveal that flamingos are not monogamous as long …

The present study focused on methylmercury (MeHg) in emergent aquatic insects and spiders from human-made ponds. This dissertation addresses two main topics: (1) factors affecting variation in spider MeHg concentrations around human-made ponds and (2) the magnitude of MeHg transported out of human-made ponds by emergent aquatic insects (insect-mediated MeHg flux). Spiders were specifically targeted in this study because they have been proposed as sentinels of MeHg contamination (organism whose tissue concentrations reflect the level of MeHg in the environment). Spider MeHg concentrations were related to spider diet, size, and proximity to waterbody, but affected individual spider taxa differently. In a second study, I found that only "large" spiders within a taxa had tissue concentrations positively related to prey MeHg concentrations. These results indicate that the relationship between spider and prey MeHg could be size-dependent and that "large" spiders within a taxa may better reflect ambient MeHg contamination. Finally, I tested a conceptual model hypothesizing insect-mediated MeHg flux from human-made ponds is controlled by pond permanence and fish presence. In agreement with the conceptual model, insect-mediated MeHg flux from ponds was suppressed by the presence of fish, likely due to fish predation on emergent insect larvae. I found the mean …

Networks of ground-based and satellite borne instruments to measure ultraviolet (UV) sunlight and total column ozone have greatly contributed to an understanding of increased amounts of UV reaching the surface of the Earth caused by stratospheric ozone depletion. Increased UV radiation has important potential effects on human health, and agricultural and ecological systems. Observations from these networks make it possible to monitor total ozone decreases and to predict ozone recovery trends due to global efforts to curb the use of products releasing chemicals harmful to the ozone layer. Thus, continued and expanded global monitoring of ozone and UV is needed. However, existing automatic stratospheric ozone monitors are complex and expensive instruments. The main objective of this research was the development of a low-cost fully automated total column ozone monitoring instrument which, because of its affordability, will increase the number of instruments available for ground-based observations. The new instrument is based on a high-resolution fiber optic spectrometer, coupled with fiber optics that are precisely aimed by a pan and tilt positioning mechanism and with controlling programs written in commonly available software platforms which run on a personal computer. This project makes use of novel low-cost fiber optic spectrometer technology. A cost …

The Texas Stream Team (TST) is one of an increasing number of citizen science water monitoring programs throughout the US which have been continuously collecting surface water quality data under quality assurance protocols for decades. Volunteer monitoring efforts have generated monitoring datasets that are long-term, continuous, and cover a large geographic area - characteristics shown to be valuable for scientists and professional agencies. However, citizen science data has been of limited use to researchers due to concerns about the accuracy of data collected by volunteers, and the decades of water quality monitoring data collected by TST volunteers is not widely used, if at all. A growing body of studies have attempted to address accuracy concerns by comparing volunteer data to professional data, but this has rarely been done with large-scale, existing datasets like those collected by TST. This study assesses the accuracy of the volunteer water quality data collected across the state of Texas by the TST citizen science program between 1992-2017 by comparing it to professional data from corresponding stations during the same time period, as well as comparing existing and experimental data from a local TST partner agency. The results indicate that even large-scale, existing volunteer and professional …

The physiological responses of Myriophyllum spicatum to 2,4-D, diquat and copper were quantified using a plant tissue viability assay, and daily measures of dissolved oxygen and pH. Correlations of herbicide tissue residues to physiological response measures were determined and the relationship was used to develop exposure-response models. Diquat and copper had a greater effect on plant tissue viability than was observed for 2,4-D. Diquat produced greater reductions in dissolved oxygen concentrations and pH values than 2,4-D or copper. Copper exposure had the least effect on these parameters. Exposure-response models developed for 2,4-D predicted effective control at plant tissue residues ranging from 4000 to 4700 mg/kg. Aqueous exposure concentrations necessary to produce effective control plant tissue residues ranged from 0.20 to 0.40 mg/L. Exposure-response models developed for diquat predicted effective control at plant tissue residues ranging from 225 to 280 mg/kg. Aqueous exposure concentrations necessary to produce effective control plant tissue residues ranged from 0.113 to 0.169 mg/L. Exposure-response models developed for copper predicted effective control at plant tissue residues ranging from 680 to 790 mg/kg. Aqueous exposure concentrations necessary to produce effective control plant tissue residues ranged from 0.32 to 0.64 mg/L. Model predictions for 2,4-D, diquat and copper were …

Lewisville Lake Environmental Learning Area (LLELA) is an 809-hectare property in Denton County, TX. A study of the vegetation community identified 466 species in 104 families, with 25% of the species from only two families, Asteraceae and Poaceae. The property demonstrates the characteristics of an early successional community, dominated by weedy species. Prairie communities are dominated by Johnson grass and ragweed, with climax tall grass prairie communities only in areas that have been planted with native grass seed. Forest communities are similarly in an early successional stage, dominated by the hackberry-elm-ash alliance, with small remnants of native Cross Timbers found in isolated patches. Species richness and diversity were highest in the forests and lowest in the wetlands; evenness, though not different across ecosystems, demonstrated a strong seasonal component. The species list was compared with previously reported lists for Denton County, and 256 species identified had not been previously reported for the county. A wetland restoration study was conducted to determine if there was a difference in survival and growth between potted transplants with intact root systems and bare-root transplants. Two different mesh sizes were used for protection, and the success of the different caging was evaluated. Of eight species, only …

The purpose of the research was to treat immunodeficient mice, implanted with colon-25 tumors, with continuous and interrupted electrochemical therapy (ECT). ECT involves the placement of two electrodes, an anode near the center of the tumor and a cathode into the tumor periphery. A constant voltage is applied across the electrodes for a given period of time. The data showed that the interrupted and continuous ECT resulted in a decrease in mean tumor growth as compared to that of the sham controls. The histology of both ECT groups showed an increase presence of large vacuoles, randomly distributed tumor cells as well as the presence of "crevicing" in the medullary tissue. The differential leukocyte counts showed a distinct neutrophilia and lymphopenia in all groups at day 20 post tumor implantation. The results from the experimental groups appeared to support the findings of previous investigators.

Research continues to show that pharmaceutical environmental contamination has caused adverse environmental effects, with one of the most studied effects being feminization of fish exposed to pharmaceutical endocrine disruptors. Additionally, there are also public health risks associated with pharmaceuticals because in-home reserves of medications provide opportunities for accidental poisoning and intentional medication abuse. Pharmaceutical take back programs have been seen as a remedy to these concerns; however a thorough review of peer-reviewed literature and publicly available information on these programs indicates limited research has been conducted to validate these programs as a purported solution. Furthermore, there are significant data gaps on key factors relating to take back program participants. The purpose of this dissertation was therefore to address these gaps in knowledge and ultimately determine if take back programs could actually improve public and environmental health. This was accomplished by conducting social and scientific research on a take back program called Denton Drug Disposal Day (D4). Socioeconomic, demographic, and geographic characteristics of D4 participants were investigated using surveys and geographic analysis. Impacts on public health were determined by comparing medications collected at D4 events with medications reported to the North Texas Poison Center as causing adverse drug exposures in Denton …

The role of operant behavior in the metabolism of brain 5-hydroxytryptamine (5-HT) turnover was investigated. Two and one-half hours following the administration of 150 mg/kg of para-chlorophenylalanine (PCPA), a specific inhibitor of tryptophan hydroxylase, levels of 5-HT were compared in sedentary and performing rats. Whole brain levels of serotonin were reduced in both responding and sedentary animals; however, differences between these groups were not statistically significant. The drug induced decrease in 5-HT levels was accompanied by a significant decrease in session responding. The degree of suppressed responding could be correlated with the level of 5-HT following PCPA, suggesting that the metabolism of serotonin is in part modulated by the rate of responding as maintained by the operant schedule.

The linkage relationship and arm location of the gene for greenbug resistance in the variety Will was determined by using primary trisonomics and tertiary trisomic homozygous translocations. The gene for greenbug resistance was found to be on linkage group 1 by using primary trisonomics. The gene was located on the cetromere segment of the Tl-6a translocation by using a tertiary trismoic homozygous for greenbug resistance. The data further substantiates the feasibility of using trisomics in placing genes on proper linkage groups.

Starch gel electrophoresis was used to compare 14 proteins encoded by 15 loci for seven species of the family Icteridae. A close genetic relationship among these species was classified into three groups. The Agelaiine group contained Agelaius phoeniceus, Sturnella magna, and S. neglecta. The Quiscaline group contained Euphagus cyanocephalus, Cassidix mexicanus, and Quiscalus quiscula. Molothrus ater, the most divergent, was placed in a separate group. Divergence times for the seven species were compared to the literature. Heterozygosity of the seven populations of the two species of Sturnella were compared to determine factors influencing their divergence. Two factors proposed were heterosis in S. neglecta and possible hybridization between S. neglect and S. magna.

Electrophoretic variation in 18 proteins encoded by 20 autosomal loci was used to compare the genetic relationships of 19 natural populations representing three species of the subgenus Neotoma. Of the 20 loci examined nine were monomorphic and fixed for the same allele in all populations. No more than seven loci were polymorphic within a single population. Genetic variability was expressed as the proportion of loci heterozygous in the average individual of a population. Heterozygosity in the three species of Neotoma studied averaged 0.078, a value within the range reported for other rodents. Although the levels of heterozygosity seen in Neotoma could not readily be explained, the variation may be attributed to ecological factors. The three species of Neotoma were compared on the basis of genetic similarity and found to form a close taxonomic unit, probably semispecies. Divergence times were obtained for the three species and found to compare well with divergence times obtained from fossil data. In general, the three species have diverged within the last 112,000 years during the Wisconsin glacial period.

The Supercritical Fluid Extraction (SFE) technique was used to fractionate Suwannee River reference fulvic acid (FA). The fractions were characterized by gas chromatography (GC) and reversed-phase high performance liquid chromatography (RP-HPLC). A SFE fractionation method was developed using stepwise gradient of supercritical CO₂ and methanol. Three FA fractions were separated. The average mass recovery was 102% with the coefficient of variation of 2.8%. The fractionation dynamics and the difference in the ratios of UV absorption to fluorescence emission indicate the real fractionation of the FA. The HPLC chromatographic peak patterns and the spectra of the corresponding peaks were almost indistinguishable. The overall results of this research support the argument that FA exhibits polymer-like molecular structure.

Superior swimming stamina in adult fish is presumably passed on to their offspring, but the ontogeny of the appearance of superior stamina and the requisite enhanced cardio-respiratory support for locomotion in larval fishes has not been determined. Is the expression of the suite of parental traits enabling superior swimming stamina in their offspring dependent upon their achieving juvenile/adult morphology, or does it appear earlier in their larvae? To answer this, adults were classified into three groups based on swimming stamina, followed by measurement of length, mass, and width. Larval offspring from the two parental groups -high stamina larvae (HSL) and low stamina larvae (LSL)- were reared at 27°C in aerated water (21% O2). Routine and active heart rate, routine and active mass specific oxygen consumption were recorded through 21dpf, and cost of transport (COT) and factorial aerobic scope were derived from oxygen consumption measurements. Routine heart rate at 2dpf of LSL was 164 ± 1 b·min-1, compared to only 125 ± 2 b·min-1 for HSL. Routine heart rate subsequently peaked at 203 ± 1 b·min-1 at 5dpf in the HSL group, compared to 207 ± 1 b·min-1, at 4dpf in the LSP larvae. Active heart rate at 5 dpf of …

The regulation of short chain fatty acid metabolism has been examined. Metabolism of acetoacetate, and short chain fatty acids such as butyrate and valerate, is predicated upon the expression of genes of the ato operon. Acetoacetate induces expression of a CoA transferase (encoded by the atoDA genes) and expression of a thiolase (encoded by the atoB gene). Metabolism of saturated short chain fatty acids requires the activities of the transferase and thiolase and enzymes of 6-oxidation as well. Spontaneous mutant strains were isolated that were either constitutive or that were inducible by valerate or butyrate instead of acetoacetate.

Triclosan and triclocarban are antimicrobial compounds added to a variety of consumer products that are commonly detected in waste water effluent. The focus of this study was to determine whether the bioconcentration of these compounds in wetland plants and sediments exhibited species specific and site specific differences by collecting field samples from a constructed wetland in Denton, Texas. The study showed that species-specific differences in bioconcentration exist for triclosan and triclocarban. Site-specific differences in bioconcentration were observed for triclosan and triclocarban in roots tissues and sediments. These results suggest that species selection is important for optimizing the removal of triclosan and triclocarban in constructed wetlands and raises concerns about the long term exposure of wetland ecosystems to these compounds.