Genetic variation and thermal tolerance were surveyed for variation attributed to nonuniform selection pressures for five populations of the red shiner, Notropis lutrensis, collected from regulated and unregulated portions of a Texas river. Populations within 30 km of a hypolimnion-release dam that experience large thermal perturbations were found to have higher levels of heterozygosity, higher levels of polymorphism, significantly depressed levels of upper thermal tolerance endpoints, and greater variances in tolerance endpoints. These populations have evolved enzyme systems differing from the unregulated populations in response to a variable and depressed thermal regime.

Electrophoretic variation at twenty gene loci, patterns of behavioral thermoregulation, and genotype-specific malate dehydrogenase kinetics were investigated among populations of the red shiner, Notropis lutrensis, and the blacktail shiner, N. venustus, collected from thermally altered and thermally unaltered portions of their ranges. Genic variation was found to be high among red shiners and low among blacktail shiners. The behavioral response of the blacktail shiner to temperature was fixed among the populations sampled, whereas the response of the red shiner was mutable. Finally, blacktail shiners have incorporated into their genome an Mdh-B allele which functions well at low temperatures; red shiners, displaying high levels of Mdh-B polymorphism, maintain a more complex set of allozymes which function well over a wide range of environmental temperatures. These data are consistent with reported ecotypic distributions of the species in Texas waters; i.e., blacktail shiners occur in cool, thermally static habitats, and red shiners are tolerant of wide temperature ranges.

Asparate transcarbamoylase catalyzes the first committed step in the de novo synthesis of pyrmidine nucleotides UMP, UDP, UTP, and CTP. The archetype enzyme found in Escherichia coli (310 kDa) exhibits sigmodial substrate binding kinetics with positive control by ATP and negative control with CTP and UTP. The ATCase characterized in this study is from the extreme thermophilic Archaebacterium, Methanococcus jannaschii. The enzyme was very stable at elevated temperatures and possessed activity from 20 degrees Celsius to 90 degrees Celsius. M. Jannaschii ATCase retained 75% of its activity after incubation at 100 degrees Celsius for a period of 90 minutes. No sigmodial allosteric response to substrate for the enzyme was observed. Velocity substrate plots gave Michaelis-Menten (hyperbolic) kinetics. The Km for aspartate was 7 mM at 30 degrees Celsius and the KM for carbamoylphosphate was .125 mM. The enzyme from M. jannaschii had a broad pH response with an optimum above pH 9. Kinetic measurements were significantly affected by changes in pH and temperature. The enzyme catalyzed reaction had an energy of activation of 10,300 calories per mole. ATCase from M. jannaschii was partially purified. The enzyme was shown to have a molecular weight of 110,000 Da., with a subunit molecular …

The purpose of this study was to investigate the effects of Pb and hypoxia co-exposure on Pb toxicity and Ca homeostasis in early life stage (ELS) zebrafish (Danio rerio). Previous evidence indicates that exposure of ELS zebrafish to hypoxia (~20% air saturation) reduces Ca uptake, likely through down-regulation of the apical epithelial Ca channel (ECaC). Considering that Pb and Ca are known antagonists and compete for uptake pathways, it was hypothesized that co-exposure of Pb with hypoxia would decrease Pb toxicity by reducing Pb uptake (likely mediated through a reduced number of ECaCs). However, it was shown that at 96 hpf, whole body accumulation of both Pb and Ca was lower at 40% air saturation compared to 100% and 20% air saturation. This result closely aligned with the 96h LC50 results which showed the highest mortality of zebrafish at 40% compared to the other air saturation levels. This suggests that toxicity is likely the result of exacerbated hypocalcemia at 40% air saturation due to both Pb competition for Ca binding to Ca uptake channels/transporters, such as ECaC, and potentially reduced expression of such channels/transporters in response to this level of hypoxia. Overall, it appears that ELS zebrafish respond differentially to …

Reptiles can often experience perturbations that greatly influence their metabolic status (e.g., temperature, exercise, digestion, and ontogeny). The most common cause of fluctuations in metabolic status in post-embryonic reptiles is arguably digestion and physical activity (which will be further referred to as exercise). The objective of this thesis is to determine the mechanisms involved in CO2 transport during digestion, determine the mechanisms that allow for the maintenance of acid-base homeostasis during digestion, and observing the effect of an understudied form of exercise in semi-aquatic reptiles on the regulation of metabolic acidosis and base deficit. This dissertation provided evidence for potentially novel and under investigated mechanisms for acid-base homeostasis (e.g., small intestine and tissue buffering capacity; Chapters 3 & 4), while also debunking a proposed hypothesis for the function of an anatomical feature that still remains a mystery to comparative physiologist (Chapter 2). This thesis is far from systematic and exhaustive in its approach, however, the work accomplished in this dissertation has become the foundation for multiple distinct paths for ecologically relevant investigations of the regulation of metabolic acidosis/alkalosis in reptiles.

There has been a rapid surge toward "organic" products devoid of GMOs, MSGs, and other common compounds found in processed foods that continue to indicate an association with an increased risk for disease. These consumers seek nutrients and vitamins that are lacking in their diet and lifestyle in the form of nutraceuticals for disease prevention and treatment as well as overall lifestyle enhancement. However, these products generally lack clinical evidence as well as legal definition. Due to this ambiguity, nutraceuticals are neither considered a food product nor a pharmaceutical product. Furthermore, due to their alleged natural properties allowing for safe, therapeutic effects, nutraceuticals are being eagerly sought after by consumers in the place of pharmaceuticals. Additionally, since nutraceutical substances are "naturally" derived, there is a general lack of regulation regarding the manufacturing and distribution process. This mismanagement leads to lack of quality assurance (QA) and quality control (QC) protocols strictly implemented to define appropriate production and storage parameters. Without these critical measures, consumers are subjected to contamination of their products resulting from improper storage conditions and unmanaged production. These contaminants often include heavy metal impurities, pesticides, bacterial activity, and may also be adulterated with illicit drugs, all leading to detrimental …

We investigated the modifying effects of ultraviolet (UV) light and chemical dispersant (Corexit 9500A) on crude oil toxicity in juvenile mysids (≤ 24 h) (Americamysis bahia) and larval red drum (24-72 hpf) (Sciaenops ocellatus). These results demonstrate that crude oil toxicity significantly increases with co-exposure to environmentally relevant UV levels in both species, indicating photo-induced toxicity. This toxicity was further exacerbated by the application of chemical dispersants which increased the dissolution and concentration of oil-derived polycyclic aromatic hydrocarbons (PAHs) in test solutions. To better understand the mechanisms and initiating events of this observed photo-induced toxicity, the incidence of apoptotic cell death and global transcriptomic changes were assessed in larval red drum (24-72 hpf) following co-exposure to crude oil and UV. These results showed that co-exposure to UV and low concentrations of crude oil (<1 µg/L ∑PAH50) induced apoptotic cell death in skin and eye tissue and altered transcriptomic pathways related to visual processing and dermatological disease. To link these cellular and molecular impacts of photo-induced toxicity to apical endpoints of ecological performance, sublethal impacts to growth, metabolic rate, and visually mediated behaviors were explored in larval red drum at 2 developmental stages. These results suggested that earlier life stages may …

Pseudomonas aeruginosa is a Gram-negative rod, aerobic, non-fermenting, oxidase positive, pigment producing, and nutritionally versatile bacterium. Infections by P. aeruginosa are the most important cause of morbidity and mortality in immunocompromised patients, given virulence factor production that suppresses antibiotic therapy and promotes persistent infection. This research is the first comprehensive report of the pyrimidine biosynthetic pathway for all phases of growth in minimal and rich media coupled with the evaluation of virulence factor production of P. aeruginosa in comparison to four other bacterial species (Pseudomonas putida, Pseudomonas fluorescens, Burkholderia cepacia, and Escherichia coli wild-type strains). Cellular growth and passing genetic information to the next generation depend on the synthesis of purines and pyrimidines, the precursors of DNA and RNA. The pyrimidine biosynthetic pathway is essential and found in most organisms, with the exception of a few parasites that depend upon the pyrimidine salvage pathway for growth. Both the pyrimidine biosynthetic and salvage enzymes are targets for chemotherapeutic agents. In our laboratory, research on pyrimidine auxotrophic mutants showed the role of the pyrimidine biosynthetic pathway and its intermediates on P. aeruginosa metabolism and impaired virulence factors production. The present research shows that pyrimidine enzymes are active in all phases of growth, …