Genomic modifications occur slowly across generations, whereas short-term epigenetic transgenerational inheritance of adaptive phenotypes may be immediately beneficial to large numbers of individuals, acting as a bridge for survival when adverse environments occur. In this study we used dietary exposure to crude oil as an example of an environmental stressor to assess its effects from the molecular to the organismal levels in piscine and avian animal models. In addition, we assessed the role of the parental exposures on their offspring F1 generation. The research developed in this dissertation has contributed to several areas of investigation including molecular biology, animal physiology, and evolutionary biology. The quantitative information from these studies may be utilized to supplement information regarding the proximate and ultimate effects of environmental stressors on fish and bird populations. Furthermore, this information may be used as additional support for understanding the conservation of the responses from the molecular to the whole organismal levels across the vertebrate taxa, as well as their implications for population survival and maintenance. Additionally, the zebrafish (Danio rerio), the Siamese fighting fish (Betta splendens) and the king quail (Coturnix chinensis) have proven to be excellent models to start building a strong basis for understanding the effects …

The present study sought to determine some morphological and physiological critical windows during very early development in zebrafish exposed to crude oil. I hypothesized that exposed zebrafish would present a decrease in survival rate and body mass, and an increase in routine oxygen consumption (ṀO2), and critical oxygen tension (PCrit). To test these hypotheses, zebrafish were acutely exposed (24 h) during different days of development (1 to 6 days post-fertilization, dpf) to different concentrations of high-energy water-accommodated fractions (HEWAFs). The endpoints of survival, body mass, routine oxygen consumption, and critical oxygen partial pressure were measured at 7 dpf. Survival rate decreased based on the exposure concentration but not as a function of the day of crude oil exposure. No significant effects were found in PCrit. Body mass was reduced by the different concentrations of HEWAF, with the size of the effect varying with exposure day, with the effect strongest on when exposure occurred at 2 and 3 dpf. Oxygen consumption (ṀO2) differed significantly depending upon the day of exposure in fish exposed to crude oil. Specifically, HEWAF exposure significantly increased ṀO2 in larvae exposed at 3 dpf (9.081 µmol O2/g/h, ±0.559) versus 2 dpf (6.068 µmol O2/g/h, ±0.652) and 6 …

Cardiac, metabolic and growth response of early-stage chicken embryos to perturbations in yolk environment was investigated. Also, effects of parental hypoxia exposure on hypoxia resistance, thermal tolerance and body length of zebrafish larvae were investigated. In the first study, thyroxine, triiodothyronine and testosterone produced differential effects on heart rate and development rate of chicken embryos during the first 4 days of development. Triiodothyronine caused a dose-dependent increase in heart rate when applied at 40 or 70 hours of age, while thyroxine caused a dose-dependent increase in heart rate when applied at 40 hours only. Testosterone and propyl-thiouracil (deiodinase antagonist) did not have an effect on heart rate. Development rate was not changed by thyroxine, triiodothyronine, testosterone or propyl-thiouracil, which suggested that heart rate changes did not result from changes in embryo maturity. In the second study, chicken embryos exposed to yolks of different bird species during early-stage embryonic development showed changes in heart rate, mass-specific oxygen consumption and body mass that scaled with the egg mass, incubation period length, and yolk triiodothyronine and testosterone levels of the species from which yolk was derived. In the third study, this phenomenon was investigated between layer and broiler chickens. Heart rate, oxygen consumption …

Respiratory and metabolic patterns in response to variations in exercise workload (WL) and pedal frequency (RPM) were examined in 10 healthy males. Each subject performed WLs of low (L), moderate (M) and high (H) intensity, equivalent to 25%, 50% and 75% V02 m a x at 7 pedal frequencies (40, 50, 60, 70, 80, 90 and 100 RPM). ANOVA ( 3 X 7 design) indicated that WL and RPM had independent and significant effects on all respiratory and metabolic measures; i.e., the greater the WL and RPM, the higher the HR, V02, VC02, Ve, Fb, Vt, Vt/Ti, Vt/Te and Ti/TtQt and the lower the Ti and Te. However, analysis of the interaction effect revealed different response patterns for Fb, Vt, Ti, Vt/Ti, Vt/Te and Ve among the WLs. During L-WL, increases in RPM produced increases in Ve which were due to progressive increases in both Fb and Vt. However, during M-WL and H-WL, increases in RPM produced increases in Ve which were accomplished by a constant Vt and a progressive increase in Fb. My findings suggest that during low WLs, the signal for Vt is dependent on rate of contraction, while during M-WL and H-WL, the signal for Vt appears …

Changes in spontaneous spike activities recorded from murine frontal cortex networks grown on substrate-integrated microelectrodes were used to determine the dissociation constant (KB) of three GABAA antagonists. Neuronal networks were treated with fixed concentrations of GABAA antagonists and titrated with muscimol, a GABAA receptor agonist. Muscimol decreased spike activity in a concentration dependent manner with full efficacy (100% spike inhibition) and a 50% inhibitory concentration (IC50) of 0.14 ± 0.05 µM (mean ± SD, n=6). At 10, 20, 40 and 80 µM bicuculline, the muscimol IC50 values were shifted to 4.3 ± 1.8 µM (n=6), 6.8 ± 1.7 µM (n=6), 19.3 ± 3.54 µM (n=10) and 43.5 µM (n=2), respectively (mean ± SD). Muscimol titration in the presence of 10, 20, 40 µM of gabazine resulted in IC50s values of 20.1 (n=2), 37.17 (n=4), and 120.45 (n=2), respectively. In the presence of 20, 80, and 160 µM of TMPP (trimethylolpropane phosphate) the IC50s were 0.86 (n=2), 3.07 (n=3), 6.67 (n=2) µM, respectively. Increasing concentrations of GABAA antagonists shifted agonist log concentration-response curves to the right with identical efficacies, indicating direct competition for the GABAA receptor. A Schild plot analysis with linear regression resulted in slopes of 1.18 ± 0.18, 1.29 …

The soil-nematode Caenorhabditis elegans survives oxygen deprivation (anoxia < 0.001 kPa of O2, 0% O2) by entering into a state of suspended animation during which cell cycle progression at interphase, prophase and metaphase stage of mitosis is arrested. I conducted cell biological characterization of embryos exposed to various anoxia exposure times, to demonstrate the requirement and functional role of spindle checkpoint gene san-1 during brief anoxia exposure. I conducted a synthetic lethal screen, which has identified genetic interactions between san-1, other spindle checkpoint genes, and the kinetochore gene hcp-1. Furthermore, I investigated the genetic and cellular mechanisms involved in anoxia-induced prophase arrest, a hallmark of which includes chromosomes docked at the nuclear membrane. First, I conducted in vivo analysis of embryos carried inside the uterus of an adult and exposed to anoxic conditions. These studies demonstrated that anoxia exposure prevents nuclear envelope breakdown (NEBD) in prophase blastomeres. Second, I exposed C. elegans embryos to other conditions of mitotic stress such as microtubule depolymerizing agent nocodazole and mitochondrial inhibitor sodium azide. Results demonstrate that NEBD and chromosome docking are independent of microtubule function. Additionally, unlike anoxia, exposure to sodium azide causes chromosome docking in prophase blastomeres but severely affects embryonic viability. …

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental toxicants found in anthropogenic mixtures such as crude oil, air pollution, vehicle exhaust, and in some natural combustion reactions. Single PAHs such as benzo[a]pyrene (BaP) also impact fish behavior when animals are exposed in early life stages and for short periods of time. Aquatic animals such as fish may encounter BaP through road runoff and oil spills, but few studies have examined the impact of aqueous exposure on adult fish, and fewer have examined the resulting fitness-relevant behavioral consequences of BaP and PAH mixtures and their long-term persistence. This dissertation targets this gap in the literature by examining how aqueous exposure to BaP influences anxiety-like behavior, learning, and memory in adult zebrafish, and how parental exposure to the PAH mixture, crude oil, combined with hypoxia affects social and exploratory behavior in unexposed larval zebrafish. We found that learning and memory were not affected by 24 hour exposure to BaP, that anxiety-like behavior was minimally affected, and that locomotor parameters such as distance moved and times spent in darting and immobile states were significantly altered by exposure to BaP. Additionally, we found that parental exposure to crude oil and hypoxia decreased larval velocity. Additionally, …

Factors currently at the forefront of human in vitro fertilization (IVF) that collectively influence treatment success in the form of blastocysts development were investigated during early mammalian embryology with concentration on infertile patients presenting with diminished ovarian reserve or preliminary ovarian failure. A novel experimental technique, Graft Transplant-Embryonic Stem Cells (GT-ESC) was introduced in the mouse model, as the first inclusive approach for embryo selection in IVF treatments resulting in successful graft integration of sibling cells, stage-dependent (day 4) blastocysts. E-Cadherin-catenin bonds play an integral role in trophectoderm cell viability and calcium removal, inducing disruption of cell-to-cell bonds at the blastocyst stage was detrimental to continued blastocyst development. One of the leading methods for embryo selection for uterine transfer in human IVF is application of pre-implantation genetic screening (PGS) methods such as next generation sequencing (NGS). Female patients <35 y do not benefit from this treatment when outcome is measured by presence of fetal heart beats at 10 weeks of gestation. Patients 35-37 y benefit from PGS with no significant difference of outcome based on form of PGS method utilized. Therefore, small nucleotide polymorphism array (snp-array) or targeted-NGS should be selected for this age range to lessen the financial burden …

To evaluate the blood-brain barrier system in the pathogenesis of an irreversible hypothermic state in dogs, concentrations of 2 4 Na and 86Rb were measured at body temperatures ranging from 37 0 C to 160 C. A suppression of transport of sodium was demonstrated, followed by an increase as the temperature was lowered. The concentration of rubidium ion increased in concentration as the temperature fell. These data indicate there may be a temperature threshold below which the blood-brain barrier system fails to maintain the internal environment of the central nervous system. The intimate relationship of several brain stem nuclei with the cerebro-spinal fluid indicates they may be at risk during profound cooling.

The Deepwater Horizon oil drilling rig exploded on April 20, 2010, off the coast of Louisiana in the Gulf of Mexico. Following the spill, British Petroleum, leaser of the rig, set up a funding institution known as the Gulf of Mexico Research Initiative (GoMRI) to support research and understanding of the spill on the environments and peoples of the gulf. This outreach project was created alongside research of the RECOVER consortium, funded by GoMRI, to communicate what is happening within research labs around the country to understand the effect that the spill had on fish in pelagic and coastal regions of the gulf. The outreach project is composed of a short film (Deepwaters: The Science of a Spill, 18 min) and related outreach materials posted to Instagram (@FishandOilSpills).

The Deepwater Horizon oil spill released millions of barrels of oil into the Gulf of Mexico, impacting economically and ecologically important fishes. Polycyclic aromatic hydrocarbons (PAHs) present in the oil have been shown to cause developmental impairments in early life stage fishes, such as morphological and behavioral changes related to eye formation and visual processing following PAH exposure. Prior research reported reduced eye growth in open water, pelagic species, as well as reduced photoreceptor-specific transcription factors associated with eye development following exposure to crude oil. Though changes in transcriptomic-level pathways associated with vision and visual processing have been reported, it has yet to be determined how these changes relate to physiological or behavioral-level effects in fish. Therefore, the present studies evaluated the effect of weathered crude oil on eye development and visual function in mahi-mahi, red drum, sheepshead minnow, and zebrafish larvae. Fish were assessed through several visually-mediated behavioral assays, analyzed histologically and immunohistologically, along with subsequent transcriptomic analyses and associated gene expression changes. Larvae exposed to crude oil experienced significantly reduced abilities to exhibit optomotor or optokinetic responses relative to controls, with associated reductions in retinal development. Furthermore, genes associated with eye development and phototransduction were downregulated, with subsequent …

Oxygen deprivation can be pathological for many organisms, including humans. Consequently, there are several biologically and economically relevant negative impacts associated with oxygen deprivation. Developing an understanding of which genes can influence survival of oxygen deprivation will enable the formulation of more effective policies and practices. In this dissertation, genes that influence adult anoxia survival in the model metazoan system, C. elegans, are identified and characterized. Insulin-like signaling, gonad function and gender have been shown to influence longevity and stress resistance in the soil nematode, C. elegans. Thus, either of these two processes or gender may influence anoxia survival. The hypothesis that insulin-like signaling alters anoxia survival in C. elegans is tested in Aim I. The hypotheses that gonad function or gender modulates anoxia survival are tested in Aim II. Insulin-like signaling affects anoxia survival in C. elegans. Reduction of insulin-like signaling through mutation of the insulin-like receptor, DAF-2, increases anoxia survival rates in a gpd-2/3 dependent manner. The glycolytic genes gpd-2/3 are necessary for wild-type response to anoxia, and sufficient for increasing anoxia survival through overexpression. Gonad function and gender both affect anoxia survival in C. elegans. A reduction of ovulation and oocyte maturation, as measured by oocyte flux, …

A prospective ecological risk assessment method was developed evaluating the cumulative probabilistic impact of chemical stressors to aquatic organisms. This method was developed in response to the need to evaluate the magnitude, duration and episodic nature of chemical stressors on aquatic communities under environmental exposure scenarios. The method generates a probabilistic expression of the percent of an ecosystem's species at risk from a designated chemical exposure scenario.

Sensory neocortex contains a significant number of inhibitory neurons that use gamma-aminobutyric acid (GABA) as their neurotransmitter. Functional roles for these neurons have been identified in physiological studies. For example, in primary somatosensory cortex (SI), blockade of GABAa receptors with bicuculline leads to expansion of receptive fields (RFs). The magnitude of RF enlargement varies between SIpopulations of GABAergic neurons were identified by labeling specific calcium binding proteins.

Small isolated populations are of particular conservation interest due to their increased extinction risk. This dissertation investigates two small wild bird populations using genetic approaches to inform their conservation. Specifically, one case study investigated a Greater Sage-grouse (Centrocercus urophasianus) population located in northwest Wyoming near Jackson Hole and Grand Teton National Park. Microsatellite data showed that the Jackson sage-grouse population possessed significantly reduced levels of neutral genetic diversity and was isolated from other Wyoming populations. Analysis with single nucleotide polymorphisms (SNPs) and microsatellite data provided further evidence that the population's timing of isolation was relatively recent and most likely due to recent anthropogenic habitat changes. Conservation recommendations include maintaining or increasing the population's current size and reestablishing gene flow with the nearest large population. The second case study investigated the genetic distinctiveness of the Floreana island population of the Galápagos Short-eared Owl (Asio flammeus galapagoensis). Mitochondrial DNA sequence data did not detect differences across nine island populations, yet microsatellite and morphometric data indicated that limited gene flow existed with the population and surrounding island populations, which appeared asymmetric in direction from Floreana to Santa Cruz with no indication of gene flow into Floreana. These results have important conservation implications and …

This study conducts watershed analysis using biological and geo-spatial techniques. Incorporating landscape features with biological attributes has been shown to be an effective method of monitoring environmental quality within watersheds. In situ biomonitoring using the Asiatic Clam, Corbicula fluminea, habitat suitability, and water quality data were evaluated for their potential to describe ecological conditions in agricultural and urban areas within the Upper Trinity River watershed. These data were analyzed with GIS to identify effects of land use on ecological conditions. C. fluminea downstream of point source effluents was effective detecting in-stream toxicity. Ambient toxicity appears to have improved in the Trinity, although urban influences limit aspects of aquatic life. No association between habitat quality and land use was identified.

Bacillus subtilis (ATCC 19659) were damaged by exposure to various concentrations of glutaraldehyde, as shown by decreased germination rates. The damage caused was repaired or otherwise obviated by the presence of sodium lactate in the holding medium. When two different salts of lactic acid were compared for ability to overcome the effect of glutaraldehyde, it was found that calcium salt of lactate was more effective than the sodium salt. The damage repair system involved l-alanine, lactate and either the sodium or calcium ions. The study involved in determining the difference in efficiency of spore repair was due to an organic or an amino acii snowed that the presence of two carboxylic functional groups did not effectively alter the reactivity.

My thesis focuses on the conservation consequences of the hybridization of Lesser Prairie-Chickens in Kansas. Specifically, examining how past land management practices altering the species ranges impact the distinctiveness of Lesser Prairie-Chickens. Each chapter is an individual publication that addresses if the Greater and Lesser Prairie-Chicken are distinct when applying the morphological and biological species concepts. Chapter 2 compares the evolutionary history and morphological construct of Lesser Prairie-Chickens and other Galliformes using morphometric analysis. Chapter 3 uses low-resolution microsatellite data to reflect recent changes at the population level. This study aims to observe the Greater and Lesser Prairie-Chicken using the morphological and biological species concepts, two of the many species concepts, to determine the distinctiveness and rate of hybridization for these closely related species.

In this dissertation, I explored the effects of developmental hypoxia on heart contractility in three separate species of ectotherms: the common snapping turtle (Chelydra serpentina), the American alligator (Alligator mississippiensis), and the zebrafish (Danio rerio). I began with the common snapping turtle and tested whether the utilization of the sarcoplasmic reticulum was altered in response to developmental hypoxia. In the next two chapters, developmental hypoxia of the American alligator was explored studying how the cardiac tissue was affected, specifically in physiological stressors, sarcoplasmic reticulum utilization and sensitivity to pharmacological increases in contractility. The last chapter explored how zebrafish heart contractility was altered in response to chronic hypoxia from egg to adult. Findings from these chapters suggest that while developmental hypoxia did alter cardiac contractility, it did not alter the response of the heart to physiological stressors such as increased heart rate or under hypoxia. Overall, these findings contribute to increasing the current understanding of how developmental hypoxia alters the cardiovascular system but with an emphasis on the cardiac tissue level.

Research efforts in aquatic toxicology have historically centered on the chemical analyses and toxic effects of waters to aquatic organisms. More recently, sediment-source toxicity has been explored, with efforts concentrated on establishing sensitive and accurate methodologies. This study focused on the toxicity of trivalent chromium, hexavalent chromium, and fluoranthene to Pimephales promelas, Ictalurus punctatus, and Lepomis macrochirus. Test fish were exposed to both water-borne and sediment-source toxicants for 96 hours (h) and 30 days (d). A 96-h and 30-d LC50 (mg/L Cr, ug/L Fluoranthene) was determined for each fish species exposed to aqueous toxicants. In addition, 96-h and 30-d LC50s were determined for each fish species from sediment chromium concentrations (mg/kg) and sediment fluoranthene concentrations (ug/kg). Although lethality endpoints were used throughout this research, acute effects other than mortality were determined for Lepomis macrochirus exposed to hexavalent chromium. Lethal toxicity values (96-h and 30-d LC50 and their 95% confidence limits) for trivalent chromium could not be determined since trivalent chromium concentations above 6.0 mg/L could not be obtained at water pHs compatible with these fish species. Trivalent chromium addition to test waters at pHs compatible with fish survival resulted in a chromium precipitate that was not lethal to test fish. …

Article that provides a brief overview of current research on synthetic biology and metabolic engineering approaches to control phenylpropanoid synthesis and phenylpropanoid-related biosensors, advocating for the use of biosensors and genetic circuits as a step forward in plant synthetic biology to develop autonomously-controlled phenylpropanoid-producing plant biofactories.

Organisms synthesize stress proteins in response to a variety of stressors. The 68/70-kDa proteins (synonymous to the 72/73-kDa proteins) have shown to be the most promising stress proteins, and have been proposed as a biomarker of general organismal stress. The 68/70-kDa proteins were used in an antigen/antibody based approach to determine the duration of the stress protein response of Pimephales promelas following an acute exposure to copper sulphate.

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 …

Baroreflex function and cardiovascular responses to lower body negative pressure during selective autonomic blockade were evaluated in endurance exercise trained (ET) and untrained (UT) men. Baroreflex function was evaluated using a progressive intravenous infusion of phenylephrine HCL (PE) to a maximum of 0.12 mg/min. Heart rate, arterial blood pressure, cardiac output and forearm blood flow were measured at each infusion rate of PE. The reduction in forearm blood flow and concomitant rise in forearm vascular resistance was the same for each subject group. However, the heart rate decreases per unit increase of systolic or mean blood pressure were significantly (P<.05) less in the ET subjects (0.91 ± 0.30 versus 1.62 ± 0.28 for UT). During progressive lower body negative pressure with no drug intervention, the ET subjects had a significantly (P<.05) greater fall in systolic blood pressure (33.8 ± 4.8 torr versus 16.7 ± 3.9 torr). However, the change in forearm blood flow or resistance was not significantly different between groups. Blockade of parasympathetic receptors with atropine (0.04 mg/kg) eliminated the differences in response to lower body negative pressure. Blockade of cardiac sympathetic receptors with metoprolol (0.02 mg/kg) did not affect the differences observed during the control test. It was …