Water, sediments, floodplain soils, and fish from Big and Little Bayou creeks were assessed for polychlorinated biphenyls (PCBs) and metals in 1987-2005. The streams were impacted by effluents from the Paducah Gaseous Diffusion Plant (PGDP). PCBs were rarely detected in water samples. Aroclor 1248 detections in sediments and floodplains were sporadic, but 1254 and 1260 were consistent. Seasonal variations were only observed for Aroclor 1248. Detection of PCBs in floodplains indicated recent influx of contaminated sediments. Species-specific PCB residues patterns were studied for Lepomis cyanellus, L. megalotis, L. macrochirus, and Campostoma anomalum. Highest PCB concentrations were found in stoneroller minnows. PCBs decreased over time in sunfish species, but retained a baseline level. No relationships were found between sunfish age and PCB concentrations. At low PCB levels, green sunfish body burden (BB) did not correlate with lipid content. A certain PCB concentration must be exceeded before PCB BB correlates with lipid content. Stream flows differed by season, and fish PCB BB differed by season, but stream flow did not correlate with fish BB. Congener groups corresponded to Aroclor concentrations. Higher chlorinated biphenyls peaked in the spring for sport fish. PCBs remobilized in the spring resulted in higher summer BB for the stoneroller minnows, whereas sunfish eliminated PCBs by summer. This variability was due to the sunfishs ability to regulate PCBs. Metal concentrations did not display the seasonal variability evident in the PCB data. Water metals may have peaked during storm events and decreased rapidly. Water Pb increased with time, but the source could not determined. Zinc in Little Bayou creek likely originated from the PGDP. Except for Cu and Zn, stoneroller minnow metal BB decreased with time. Metal BB were used to determine bioavailable fractions. Less than half of the water column Cd, Cr, Fe, and Pb in impacted areas were bioavailable to stoneroller minnows, whereas 59% of Ag, 73% of Cu, and 64% of Zn were bioavailable. Cumulative criterion units were calculated to determine additive metal impacts, and proved useful in locating sectors with possible ecological impact. In-situ monitors proved valuable in studying PCB and metal behavior under real-world conditions.

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.

Postpartum depression (PPD) is a major health problem that affects many women worldwide. In Egypt, PPD is neglected despite the expected high prevalence rate among women during the transition period after the Egyptian revolution. This research investigated the prevalence, risk factors, and interventions of postpartum depression in southern rural Egypt. Interviews were conducted with 57 participants recruited from public and private hospitals. Questionnaires and the Arabic version of the Edinburgh Postnatal Depression Scale were administered. The prevalence of PPD is 73.7%. PPD is associated with low income and age at childbirth. Most participants regarded screening mothers after childbirth for PPD as effective; in comparison to, antidepressants that were regarded by most participants as ineffective. Women in southern rural Egypt prefer high number of pregnancies, so investigating the influence of sertraline, an antidepressant medication, on female hormones becomes important. In this research, fathead minnows were exposed to 3 and 10 ppb sertraline for 7 days. Real-time Polymerase Chain Reaction was used to detect the change in gene expression of the Follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Results showed that a down regulation at the 10 ppb was evident on the LH and to a lesser extent on FSH. Our results increased levels of sertraline inhibited GnRH which influenced expression of LH and FSH.

Fishes in the superorder Ostariophysi, including fathead minnows (Pimephales promelas), possess specialized epidermal club cells that contain an alarm substance. Damage to these cells, as would occur during a predator attack, causes the release of the alarm substance and can indicate the presence of actively foraging predators to nearby conspecifics. For nearly 70 years, research involving epidermal club cells has focused on the alarm substance and the role it plays in predator/prey interactions. However, recent studies have indicated that there may be a connection between epidermal club cells and the fish immune system. Fish increase investment in epidermal club cells upon exposure to skin penetrating pathogens and parasites. In this study I tested for differences in epidermal club cell investment by fathead minnows exposed to the immunosuppressive effects of the glucocorticoid hormone cortisol. In experiment 1, fathead minnows were exposed to either a single intraperitoneal injection of corn oil or no injection at all. The purpose of this experiment was to determine whether corn oil, the vehicle for cortisol injections in later experiments, had an effect on epidermal club cell density. The treatments had no effect on epidermal club cell size, cell area, or epidermal thickness. In experiment 2, skin extract was prepared from the skin of corn oil injected and non injected fathead minnows as in experiment 1 to determine whether corn oil had an effect on the epidermal club cell alarm substance concentration. The treatments showed no significant differences in observed anti-predator behaviour, including change in shelter use, dashing and freezing. In experiment 3, fathead minnows were exposed to either a single intraperitoneal injection of cortisol or corn oil. The purpose of this experiment was to determine whether cortisol, a known immunosuppressant, had an effect on epidermal club cell investment. Fathead minnows exposed to a single cortisol injection had significantly reduced respiratory burst activity of kidney phagocytes indicating that there was suppression of the innate immune system. Furthermore, cortisol treated fathead minnows showed significantly lower numbers of epidermal club cells. The treatments had no effect on individual epidermal club cell area, epidermal thickness and serum cortisol levels after 12 days. The results from this experiment suggest that pharmacological cortisol injections in fathead minnows have a suppressive effect on the fish innate immune system. Furthermore, the findings that cortisol induced immunosuppression also influences epidermal club cell investment provides support for the hypothesis that epidermal club cells may function as part of the fish immune system.

To be successful, individuals that are susceptible to predation have to optimize the trade-offs between predator avoidance and other fitness related activities such as foraging or reproduction. One challenge for prey is to identify which species pose a threat and should be avoided, and which species should be ignored. The goal of this study was to investigate whether minnows can generalize recognition of predators and non-predators using visual cues. I conducted experiments in both clear and turbid conditions to test whether the level of turbidity affects the quality of visual information available to the prey and hence the ability of prey to generalize. Latent inhibition and learned irrelevance are mechanisms of learning that can be used by prey to recognize stimuli as non-risky. Repeated exposure to an unknown stimulus in the absence of risk leads to the stimulus being categorized as non-risky. Fathead minnows were pre-exposed to the sight of brook trout or control water to provide minnows the opportunity to learn to recognize the trout as a non-predator. Following this the fish were conditioned with alarm cues (AC) to the sight of each predator paired and then their responses to the sight of brook trout, rainbow trout, and yellow perch were tested either in clear or turbid water. In clear water, minnows conditioned to recognize one of the trout species generalized their response to the other species. However, when the minnows were pre-exposed to the sight of a brook trout, they were inhibited from subsequently recognizing the sight of brook trout as threat and generalized this non-predator recognition to the sight of rainbow trout but not to yellow perch. In turbid water, however, minnows that were pre-exposed to the sight of brook trout had impaired responses to all predators while those pre-exposed to water showed an intermediate intensity anti-predator response toward each predator. Overall, my results demonstrate that minnows were able to distinguish between predators and non-predators in the clear environment but turbidity influences the visual information used by minnows and hence impaired the minnow’s ability to recognize and generalize the sight of predators and non-predator species.

Fishes in the superorder Ostariophysi, including fathead minnows (Pimephales promelas), possess specialized epidermal club cells that contain an alarm substance. Damage to these cells, as would occur during a predator attack, causes the release of the alarm substance and can indicate the presence of actively foraging predators to nearby conspecifics. For nearly 70 years, research involving epidermal club cells has focused on the alarm substance and the role it plays in predator/prey interactions. However, recent studies have indicated that there may be a connection between epidermal club cells and the fish immune system. Fish increase investment in epidermal club cells upon exposure to skin penetrating pathogens and parasites. In this study I tested for differences in epidermal club cell investment by fathead minnows exposed to the immunosuppressive effects of the glucocorticoid hormone cortisol. In experiment 1, fathead minnows were exposed to either a single intraperitoneal injection of corn oil or no injection at all. The purpose of this experiment was to determine whether corn oil, the vehicle for cortisol injections in later experiments, had an effect on epidermal club cell density. The treatments had no effect on epidermal club cell size, cell area, or epidermal thickness. In experiment 2, skin extract was prepared from the skin of corn oil injected and non injected fathead minnows as in experiment 1 to determine whether corn oil had an effect on the epidermal club cell alarm substance concentration. The treatments showed no significant differences in observed anti-predator behaviour, including change in shelter use, dashing and freezing. In experiment 3, fathead minnows were exposed to either a single intraperitoneal injection of cortisol or corn oil. The purpose of this experiment was to determine whether cortisol, a known immunosuppressant, had an effect on epidermal club cell investment. Fathead minnows exposed to a single cortisol injection had significantly reduced respiratory burst activity of kidney phagocytes indicating that there was suppression of the innate immune system. Furthermore, cortisol treated fathead minnows showed significantly lower numbers of epidermal club cells. The treatments had no effect on individual epidermal club cell area, epidermal thickness and serum cortisol levels after 12 days. The results from this experiment suggest that pharmacological cortisol injections in fathead minnows have a suppressive effect on the fish innate immune system. Furthermore, the findings that cortisol induced immunosuppression also influences epidermal club cell investment provides support for the hypothesis that epidermal club cells may function as part of the fish immune system.

Harmful algal blooms (HABs) of the brevetoxin-producing dinoflagellate Karenia brevis occur periodically along the central west coast of Florida. Mass mortalities of marine birds have long been associated with these blooms, yet there is little data documenting the accumulation of brevetoxins in the tissues of birds and their prey items. An intense HAB event impacted the region from Tampa Bay to Charlotte Harbor during most of 2005. More than one hundred marine birds, representing twenty three species, were collected during this bloom. All birds sampled were found dead or had died within 24 hours of admittance to local wildlife rehabilitation centers. In order to determine if fish were vectors for brevetoxin ingestion, the stomach contents of all birds were examined and any recovered fish were identified to the extent possible. The gastrointestinal tissues and contents from all avian samples were analyzed for brevetoxin levels, with results ranging from Shorebirds and gulls may also be exposed to brevetoxins via scavenging of red tide-killed fish deposited on beaches during blooms. Samples from scavenged fish were found to have brevetoxin levels ranging from 31 to 95,753 ng PbTx per gram tissue.

Relationships between fish assemblage composition and substrate variation is poorly understood in large rivers. Information on fishes occurrence and behavior and substrate variation were examined in field observations for the Middle Wabash River and fine scale artificial streams experiments. The results from field observations suggested strong concordance for variation in abundance of fishes with habitat variation among sites, resulting in a longitudinal river gradient as dominant in the Middle Wabash River. In addition, shifts in fish behavior within artificial stream experiments demonstrated that species-specific habitat selection behaviors were influenced by interactions within a fish assemblage. The combination of artificial stream experiments and field observations can identify fine scale trends that bioassessment surveys cannot tease apart, and highlighting the need to examine species-habitat relationships at more than one scale. / Department of Biology

A number of widely used industrial chemicals have been shown to possess endocrine-disrupting properties. In this thesis, a series of in vitro tests, and an in vivo reproductive performance test with fathead minnows, were used to clarify the extent of estrogenic activity exhibited by the phthalate esters - a class of compound hitherto referred to as 'estrogenic'. Using a recombinant yeast estrogen screen, I demonstrated that a small number of commercially available phthalates showed extremely weak estrogenic activity. The most potently estrogenic phthalate of those tested was BBP, which was approximately one million-fold less potent than 17B-estradiol. The phthalates which were estrogenic in the yeast screen were also mitogenic on estrogen-responsive human breast cancer cells (MCF-7 and ZR-75 cell lines). The most prolifically used phthalate, DEHP, was not estrogenic in any of these assays. The small number of metabolites of phthalate metabolites tested (including MBuP, MBzP, MEHP and MnOP) were also not estrogenic in the recombinant yeast assay. The ability of BBP (as the most potently estrogenic phthalate in vitro) to induce a vitellogenic response (an indicator of estrogen exposure) in fathead minnows (Pimephales promelas) exposed via the water was assessed. No induction of vitellogenin was observed, indicating that 100 Jlg BBP/L (a concentration higher than would normally be found in the environment) is not estrogenic to this species of fish under the conditions employed for this experiment. In the same study, fecundity of breeding pairs of fathead minnows was assessed; exposure to BBP was not found to affect reproductive performance in these fish. A possible alternative mechanism of action of the way in which the phthalates induce frequently reported reproductive disorders was observed. Some of the phthalates, and, notably, some of their metabolites, were demonstrated to act as anti-androgens in a recombinant yeast androgen assay. 4-Nonylphenol is another industrial chemical which is used in large volumes, and due to the nature of its use (mainly in detergent formulations), is discharged into water systems via sewage effluents. This chemical has been shown to be estrogenic to fish at the concentrations at which it has been detected in the environment. 4-Nonylphenol was tested for its ability to affect plasma and pituitary gonadotropin levels in female recrudescing rainbow trout (Oncorhynchus mykiss). Plasma and pituitary levels of FSH were suppressed in fish exposed to 10 and 100 Jlg 4-NP/L. In addition, FSH gene expression was reduced in these fish, and also in the fish exposed to 1 Jlg 4-NP/L. Pituitary LH content and gene expression of this hormone were suppressed in the fish exposed to 100-, and 10- and 100 Jlg 4-NP/L respectively. Gonadal development in vertebrates is regulated by FSH. Ovarian development ceased in the fish exposed to 100 JAg 4-NP/L, possibly as a result of the suppression of FSH synthesis and/or release in these fish.

Twenty-two natural populations of the red shiner minnow, Notropis lutrensis were examined for morphological and genetic variation. This research was aimed at testing the hypothesis that morphological and genetic variation was primarily influenced by the degree of gene flow between populations. Ten linear measurements were taken from each of 1320 specimens. Morphological characters were adjusted for differential growth by least squares linear regression techniques. Genetic variability was estimated for each individual red shiner through the methods of starch gel electrophoresis. Twenty presumtive gene loci were resolved.