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Cardiovascular Fetal Programming in Quail (Colinus virginianus), An Avian Comparative Model
The consequences of early embryonic insults and how they affect subsequent life reflects the emerging concept of "fetal programming". The aim of this project is to study the effects of embryonic insults as they subsequently manifest themselves in adults, with emphasis on the heart and vasculature. My experiments establish that fetal programming operates on the bobwhite quail inducing similar changes as those observed in mammalians and other birds. The quail's fast development provides reliable data in a short period of time than other avian models (e.g. domestic chicken). Data on quail showed a correlation between egg mass and hatchling mass; where small eggs produce small hatchlings but a high mortality made it impractical as a stressor for this study. Hypoxia was used as a stressor during embryonic incubation, where it induced a low hatching weight in quail that was not observable in adult birds. Morphological measurements demonstrated an increased ventricular collagen content and reduced ventricular lumen in birds in adults incubated in hypoxia consistent with hypertension. The hematological analyzes showed few differences indicating organ remodeling instead of hematopoietic compensation. The assessment of vascular reactivity pointed out an impaired endothelium dependent relaxation commonly associated to hypertension in birds and mammals. Fetal programming could be a widespread response to an adverse prenatal environment in endotherms and the resulting data from this work contributes to our understanding of fetal programming in vertebrates and its long term consequences.
Impact of a Genetically Engineered Probiotic Therapy and IGF-1 Genomics in the PAHenu2 Mouse Model of PKU
Absence of functional phenylalanine hydroxylase results in phenylketonuria (PKU). Viable treatments remain few, expensive and secondary conditions such as osteopenia occur in most PKU patients. Objective 1: Given the recently described roles of gut microbes to aid host digestion, an orally administered genetically engineered probiotic as the delivery vehicle for enzyme replacement therapy was created. The engineered probiotic, pHENOMMenal, produced phenylalanine ammonia lyase with significant production of trans-cinnamate (phenylalanine cleavage product) in vitro and resulted in a reduction of 515 μM in blood phenylalanine when fed to PKU animals for 14 days (from 2307µM ± 264µM to 1792µM ± 261µM, n = 6, P < 0.05). The control probiotic produced no change in blood phenylalanine. Thus, pHENOMMenal treatment in PKU mice demonstrated engineered microbes could compensate for a metabolic deficiency of the host. Objective 2: Evaluate the PAHenu2 mouse model of PKU for a genetic discrepancy causing ocular enlargement and delayed development observed only after the PAHenu2 mutation was crossed to the C57BL/6J mouse. When compared to healthy littermates, ELISA indicated a consistent but insignificant decrease in plasma IGF-1 and an increase in ocular IGF-1 in PKU animals. SNP screening demonstrated a differential inheritance of IGF-1 alleles in healthy and PKU animals based on PAH allele inheritance. Ocular and developmental phenotypes in the PAHenu2 colony match those described in previous IGF-1 studies. Understanding the IGF-1 inheritance discrepancy will enable better osteopenia research using PAHenu2 mice and allow breeding of a healthier mouse colony for continued research. Collectively the results from this work describe a new therapeutic approach for treatment of PKU as well as a better understanding of the PAHenu2 mouse model to study this disease.
The Impact of Developmental Stress on Cardiovascular Physiology of Two Archosaur Species: American Alligator (Alligator mississippiensis) and Domestic Chicken (Gallus gallus)
Crocodilians and birds comprise sister taxa of archosaurs, the development of these vertebrates occurs within an egg case that leaves developing embryos susceptible to fluctuations in the nesting environment. Studies suggest that sub-optimal conditions alter morphological growth and cardiovascular physiology. Regulation of the cardiovascular system is immature in the subjects studied, and embryos may rely on humoral rather than neural control of the cardiovascular system. The primary focus of this dissertation was to assess regulatory mechanisms responsible for maintenance of arterial pressure and heart rate. Dehydration stress had marked effects on embryo growth, and altered baseline cardiovascular parameters, while leaving the response to humoral regulator, angiotensin II (Ang II), unaffected. However, dehydrated alligator embryos developed cholinergic tone on heart rate. Hypoxic incubated chicken embryos were reduced in embryo mass, and altered response to humoral regulatory components Ang I and adenosine in addition identifying a novel regulatory component of the cardiovascular response to acute hypoxia. Collectively, these studies add to the existing knowledge of cardiovascular physiology in embryonic archosaurs and suggest that some components of cardiovascular regulation are plastic following developmental stress.
Phototoxic Effects of Titanium Dioxide Nanoparticles on Daphnia Magna
Titanium dioxide nanoparticles (TiO2-NP) are one of the most abundantly utilized nanomaterials in the world. Studies have demonstrated the mechanism of acute toxicity in TiO2-NP to be the production of reactive oxygen species (ROS) leading to oxidative stress and mortality in exposed organisms. It has also been demonstrated that the anatase crystalline conformation is capable of catalyzing the cleavage of water molecules to further increase the concentration of ROS in the presence of ultraviolet radiation. This photoenhanced toxicity significantly lowers the toxicity threshold of TiO2-NP to environmentally relevant concentrations (ppb). The goal of this study was to determine whether dietary uptake and accumulation of TiO2-NP in the aquatic filter feeder Daphnia magna resulted in photoenhanced toxicity. D. magna and S. caprincornatum were exposed to aqueous solutions of 20ppm and 200ppm TiO2-NP for 24hrs and then transferred to clean moderately hard water. Samples were taken at various time points, dried, and TiO2 quantified using ICP-MS. Toxicity assays were run on D. magna using three TiO2-NP (20ppm, 200ppm) exposure protocols and two ultraviolet radiation treatments. The first exposure group was exposed to aqueous solutions of TiO2-NP for the duration of the test. The second exposure group was exposed to TiO2-NP for an hour and then transferred to clean water. The third exposure group was fed S. capricornatum that had been allowed to adsorb TiO2-NP. All samples were then placed in an outdoor UV exposure system and exposed to either full spectrum sunlight (with UV) or filtered sunlight (no UV). Here we show that TiO2 uptake peaked at one hour of exposure likely due to sedimentation of the particles out of suspension, thus decreasing bioavailability for the duration of the test. Interetsingly, when D. magna were moved to clean water, aqueous concentrations of TiO2 increase as a result of depuration from the gut …
Detection of Mercury Among Avian Trophic Levels at Caddo Lake and Lake Lewisville, TX
Mercury (Hg) is a globally distributed toxicant that has been shown to have negative effects on birds. in the United States, avian taxa have been shown to possess high Hg concentrations in the northeast, Great Lakes and Everglades ecosystems; however, few studies have measured avian Hg concentrations in other geographic regions. Previous studies have documented high Hg concentrations in multiple organisms in east Texas, but birds were not included in these studies. the main objective of the present study was to quantify Hg concentrations in birds in differing trophic levels at Caddo Lake and Lake Lewisville, TX. Results suggest that Hg concentrations may be high enough to negatively impact some bird taxa, particularly those at high trophic levels, residing at both Caddo Lake and Lake Lewisville.
Examining the Shade/flood Tolerance Tradeoff Hypothesis in Bottomland Herbs Through Field Study and Experimentation
While there is growing evidence that shade/flood tolerance tradeoffs may be important in distributions of bottomland hardwood trees and indications that they should apply to herbs, no studies have definitively explored this possibility. Four years of field data following historic flooding were supplemented with a greenhouse experiment designed to identify interactions congruent with tradeoffs. Fifteen bottomland species were grown in two levels of water availability and three levels of shade over 10 weeks. Results indicate responses of Fimbristylis vahlii and Ammannia robusta are consistent with tradeoffs. Modification of classical allometric responses to shade by substrate saturation indicates a potential mechanism for the tradeoff in A. robusta. Responses indicating potential for increased susceptibility to physical flooding disturbance are also discussed.
Integrating Concepts in Modern Molecular Biology into a High School Biology Curriculum
More so than any other science in the past several decades, Biology has seen an explosion of new information and monumental discoveries that have had a profound impact on much more than the science itself. Much of this has occurred at the molecular level. Many of these modern concepts, ideas, and technologies, as well as their historical context, can be easily understood and appreciated at the high school level. Moreover, it is argued here that the integration of this is critical for making biology relevant as a modern science. A contemporary high school biology curriculum should adequately reflect this newly acquired knowledge and how it has already has already begun to revolutionize medicine, agriculture, and the study of biology itself. This curriculum provides teachers with a detailed framework for integrating molecular biology into a high school biology curriculum. It is not intended to represent the curriculum for an entire academic year, but should be considered a significant component. In addition to examining key concepts and discoveries, it examines modern molecular techniques, their applications, and their relevance to science and beyond. It also provides several recommended labs and helpful protocols.
Investigation of Pyrimidine Salvage Pathways to Categorize Indigenous Soil Bacteria of Agricultural and Medical Importance and Analysis of the Pyrimidine Biosynthetic Pathway's Enzyme Properties for Correlating Cell Morphology to Function in All Phases of Growth
This dissertation comprises three parts and is presented in two chapters. Chapter 1 concerns Arthrobacter, a bacterium with an intriguing growth cycle. Whereas most bacteria exist as either a rod or coccus, this bacterium shares the rod/coccus lifestyle. It therefore seemed important to examine the growth regulatory pathways from the rod and coccus. The committed step, that catalyzed by aspartate transcarbamoylase (ATCase), in the pyrimidine biosynthetic pathway was chosen. The ATCase in Arthrobacter is like the well known Pseudomonas enzyme except that it has an active dihydroorotase (DHOase) associated. Included in Chapter 1 is the description of a microorganism, Burkholderia cepacia, whose ATCase has characteristics that are at once reminiscent of bacteria, mammals, and fungi. It differs in size or aggregation based on environmental conditions. In addition, it has an active DHOase associated with the ATCase, like Arthrobacter. B. cepacia is important both medically and for bioremediation. Since B. cepacia is resistant to most antibiotics, its unique ATCase is a prime target for inhibition. Whereas the first chapter deals with the de novo pathway to making pyrimidines, which is found mainly in the lag and log phase, Chapter 2 addresses the salvage pathway, which comes more into play during the stationary phase. This section focuses on the isolation, identification, and grouping of a number of natural soil bacteria from various soil locations. These organisms are important agriculturally, medically, and industrially. Addition of these soil isolates to poor soils has been found to improve the soil. In a previous study by D.A. Beck, the salvage schemes for a number of laboratory strains of microorganisms were determined. Nine separate classes of salvage were designated by determining the salvage enzymes present. In this study emphasis has been placed on soil bacteria, which had not previously been analyzed. A number of species of soil …
A Physiological Age-Grading System for Female Hydrellia pakistanae Deonier (Diptera: Ephydridae)
Conflicting opinions about the effectiveness of H. pakistanae as a biological control agent for hydrilla prompt researchers to find a method for assessing the fly's success. Developing a physiological age-grading system for the fly using ovarian morphology to detect changes in reproductive activity is useful for evaluating reproductive status of the fly in field populations. Changes in the appearance of follicular relics in ovaries with oviposition provide a reliable method to estimate fecundity. Characteristics of follicular relics were used to develop a system with eight physiological age classes, three nulliparous and five parous. Changes that occur in the fat body were used to assist in classification of nulliparous females or those with low egg counts.
The Use of Genetic Polymorphisms and Discriminant Analysis in Evaluating Genetic Polymorphisms as a Predictor of Population
Discriminant analysis is a procedure for identifying the relationships between qualitative criterion variables and quantitative predictor variables. Data bases of genetic polymorphisms are currently available that group such polymorphisms by ethnic origin or nationality. Such information could be useful to entities that base financial determinations upon predictions of disease or to medical researchers who wish to target prevention and treatment to population groups. While the use of genetic information to make such determinations is unlawful in states and confidentiality and privacy concerns abound, methods for human “redlining” may occur. Thus, it is necessary to investigate the efficacy of the relationship of certain genetic information to ethnicity to determine if a statistical analysis can provide information concerning such relationship. The use of the statistical technique of discriminant analysis provides a tool for examining such relationship.
Investigations of neuronal network responses to electrical stimulation in murine spinal cultures.
Spontaneous activity in neuronal networks in vitro is common and has been well documented. However, alteration of spontaneous activity in such networks via conditioning electrical stimulation has received much less experimental attention. Two different patterns of electrical stimulation were used to enhance or depress the level of spontaneous activity in spinal cord cultures. High-frequency stimulation (HFS), a method routinely shown to increase the efficacy of synaptic transmission, was employed to augment spontaneous activity. Low-frequency stimulation (LFS), the technique often applied to depress synaptic efficacy, was employed to decrease spontaneous activity. In addition, LFS was used to reverse the effect of HFS on spontaneous activity. Likewise, HFS was applied to counter the effect of LFS. Because these networks were grown on multi-microelectrode plates (MMEPs), this allowed the simultaneous stimulation of any combination of the 64 electrodes in the array. Thus, the possible differences in response to single versus multi-electrode stimulation were also addressed. Finally, test-pulses were delivered before and after the conditioning stimulation on the same stimulation electrode(s) in order to assess the change in mean evoked action potentials (MEAPs). Dissociated spinal tissue from embryonic mice was allowed to mature into self-organized networks that exhibited spontaneous bursting activity after two weeks of incubation. Spontaneous activity was monitored from up to 14 recording channels simultaneously. Although uniform responses to stimulation across all recording electrodes were rarely observed, a large majority of the recording channels had similar responses. Spontaneous activity was increased in 52% of 89 HFS trials, whereas activity was decreased in 35% of 75 LFS trials. The duration of most of these increases was less than 5 minutes. When there were substantial and long-term (> 15 min) changes in spontaneous activity, the opposing stimulation pattern successfully reversed the effect of the previous stimulation. The percent change in MEAPs following conditioning stimulation …
General Biology Lecture and Laboratory Curriculum Outline in a Two or Four-Year College
In July of 1999, I wrote to 24 Texas junior and community colleges (and one four-year institution) describing my thesis agenda of a general biology lecture and laboratory syllabus for introductory biology students. I requested the titles and authors of the general biology textbooks and laboratory manuals they were currently using, the publishers of these texts, and the edition of said texts. I then contacted publishers of the various textbooks who, in turn, directed me to the Dallas-area representatives for further inquiries. I assimilated the various authors' general biology topics into a two-semester syllabus of lecture and one semester of laboratory. The document is not a text manuscript, but an all-inclusive listing of a general biology syllabus broken down by subject.
Anatomy and Physiology Syllabus for Community Colleges
This syllabus includes both lecture notes and laboratory activities for a two-semester anatomy and physiology community college course. The syllabus is based on a 16-week semester that is comprised of a three-hour lecture section and a one-hour laboratory class each week. Both the lecture course and laboratory are necessary to fulfill the requirement for anatomy and physiology. Laboratory activities coincide with lectures to enhance understanding of each topic by providing visual and hands-on experiments for the concepts learned in the lecture. Laboratory quizzes will be given each week to help students maintain a working knowledge of the material learned in the laboratory. This course is appropriate for the typical anatomy and physiology student and should benefit both students who plan to major in biology and also those who are non-biology majors. Because subject matter in anatomy and physiology is quite difficult, the importance of attendance and good study skills is stressed.
Genetics Lecture and Laboratory Syllabus for a Junior-Level Course
The following is a complete syllabus for a college level genetics course. The syllabus contains lecture outlines and notes for each chapter, along with a list of transparencies needed. The quizzes and exams are prepared and placed at the beginning of the syllabus. The beginning of the course will consist of a lecture to introduce the students to the basics of genetics, followed by many applications of genetics. The process of cell division will be mastered by the students, as well as Mendelian genetics, quantitative genetics, chromosome mapping, and inheritance. The replication, synthesis, and organization of DNA are also discussed within the lectures. The final topics that will be covered using this syllabus are genetics of cancer and immunology and population genetics. These topics are essential for a detailed genetics course. The syllabus is written in great detail, and will require a full semester to be completed. The book used in association with this syllabus is Essentials of Genetics by William S. Klug and Michael R. Cummings.
A Comparison of Predicted and Actual Trophic Status of Lake Ray Roberts, Texas Based on Chlorophyll A
Two years before impoundment, the trophic status of Lake Ray Roberts was predicted by applying the total phosphorus input into the Organization for Economic Cooperation and Development (OECD) eutrophication model. Predicted mean summer epilimnetic (MSE) chlorophyll a of Elm Fork arm, Isle duBois arm and Main Body were in the eutrophic category of the OECD model. Observed MSE chlorophyll a two years after impoundment of Elm Fork arm, Isle duBois and Main Body had not reached their predicted means and were at the mesotrophic-eutrophic boundary of the OECD model. Six years after impoundment, observed MSE chlorophyll a for Main Body, was closer to its predicted mean and in the eutrophic category of the OECD model. Six years after impoundment, Elm Fork arm was the most productive area of Lake Ray Roberts. Observed means of chlorophyll a, total phosphates, suspended solids and turbidity were often highest in the Elm Fork arm. Wastewater effluent from Gainesville and Valley View, TX, had an impact on productivity in Elm Fork arm.
The Effects of Organic Surface Amendments on Soil Nutrients and Initial Tree Establishment
This study examined the effects of replicating woodland soil surface horizonation on the nutrient status of underlying soils and the initial establishment and growth of trees. A total of 283 container grown trees were planted in a bufferzone around a future landfill site. Control amendments consisted of an 8 cm layer (0.5 m3) of wood chips applied in a circular area of 4.6 m2 around the trees' planting pit. For the treatment, a 2.5 cm layer of composted biosolids (0.15 m3 or 80 Mg/ha) was applied in a circular area of 4.6 m2 around the trees' planting pit followed by an 8 cm layer (0.5 m3) of wood chips. The results indicate that the replication of woodland soil surface attributes using composted biosolids can significantly improve the nutrient status of underlying soil. Some significant effects were seen under control conditions, too. However, the effects on tree establishment and growth parameters were, for the most part, not statistically significant.
Life History and Case-building Behavior of Molanna Tryphena Betten (Trichoptera: Molannidae) in Two East Texas Spring-fed Streams
The life history and case-building behavior of Molanna tryphena from two spring-fed tributaries in East Texas were studied from January 1997 to May 1998.
Advanced Techniques in Microbial and Molecular Biology: Laboratory Procedures for a Graduate Level Course
Advanced laboratory techniques for Microbial and Molecular Biology at the graduate level are presented in this thesis. The procedures for the laboratory experiments are set forth in detail. This laboratory is conducted as two parts, each by a different professor. Part 1 covers the experiments conducted by Dr G.A.O. Donovan. These experiments include an introduction, staining procedures, biochemical reactions, mutagenesis experiment, essays,. preparation and analysis of plasmid DNA and various other topics. Part 2 covers the experiments conducted by Dr. Daniel Kunz and includes various topics like media preparation, phenotyping strains, conjugative transfer of plasmids, SDS-PAGE, induction and measurement of enzyme and transposon mutagenesis
Development of an In Vitro Protoplast Culture System for Albizia Lebek (L.) Benth., an Economically Important Leguminous Tree
An in vitro system of generating protoplasts from their callus cultures was established. The friable callus was more productive in terms of producing protoplasts than the green compact callus. The concentration of the various cell wall degrading enzymes had an effect on the viability of the protoplasts in the medium. The protoplast system developed from the experiments was stable and could be used for the transformation experiments of Albizia lebek and for other plant improvement practices.
Analysis of Phytoplankton Responses to Water Chemistry Dynamics in a Moderately Eutrophic North Texas Reservoir
Canonical correspondence analysis (CCA) was used to explore relationships between measured environmental variables and in situ phytoplankton communities in a moderately eutrophic North Texas Reservoir.
Callibaetis Floridanus (Ephemeroptera: Baetidae) Life History and Production in a West Texas Playa
A life history study of Callibaetis floridanus was conducted over the wet cycle of a playa on the Southern High Plains of Texas from June through September 1995.
Development and Application of an Assessment Protocol for Watershed Based Biomonitoring
With numerous bioassessment methodologies available, a regional protocol needs to be developed to ensure that results are comparable. A regional assessment protocol was developed that includes collecting five benthic macroinvertebrate samples, identifying organisms to genus, and calculating the following metrics: Number of Taxa, Total Number of Individuals, Simpson's Diversity Index, Shannon's Diversity Index, Percent Contribution of Dominant Taxa, Hilsenhoffs Biotic Index, and Percent Contribution of Dipterans. Once the protocol was developed, it was used to assess the Bayou Chico tributaries and watershed. All three tributaries had been significantly impacted by human activity as had the watershed as a whole. This study indicates that a regional protocol could be developed and is appropriate for biomonitoring at the watershed scale.
Light Spectra Distributions in Temperate Conifer-Forest Canopy Gaps, Oregon and in Tropical Cloud-Forest Canopy, Venezuela
Light spectra distributions were measured in two different montane forests: temperate and tropical. Spectral light measurements were made in different sized canopy gaps in the conifer forest at H. J. Andrews Experimental Forest in Oregon, USA. Researchers at Oregon State University created these gaps of 20 m, 30 m, and 50 m in diameter. In the tropical cloud forest, spectral light measurements were made in two plots that were permanently established at La Mucuy Parque Nacional in Venezuela, in collaboration with researchers at Universidad de Los Andes. In both studies, spectra and distributions of physiologically active light were analyzed: red, far-red, R/FR ratio, and blue light.
Remote Sensing of Landscape-Level Ecological Attributes at Ray Roberts Lake in North Texas
Biological diversity is a key component in assessing ecosystem health. Alteration, degradation and loss of habitat due to human influence is currently the primary stressor resulting in decreases in diversity. Reliable assessment of large areas in terms of biological integrity are needed for conservation and preservation efforts. Remotely sensed data provide an integrated view of reflected electromagnetic energy over large areas of the earth. These energy patterns provide unique spectral signatures which can be correlated to land cover and habitat. This research sought relationships between traditional ecological measures and information gathered from satellite digital imagery. Reliable interpretation of earth surface characteristics relies largely on accurate rectification to a map projection and subsequent thematic classification. Use of the Global Positioning System (GPS) for rectification was superior than digitizing topographical maps. Differentially corrected GPS locations provided optimum rectification with SPOT satellite imagery while marginally better rectifications were obtained for Landsat MSS imagery using uncorrected GPS positions. SPOT imagery provided more accurate land cover classifications than did MSS. Detection of temporal land cover change using MSS imagery was hampered by confusion among intermediate successional classes. Confusion between upland and bottomland forest classes occurred with both SPOT and MSS. Landscape analyses using thematic maps produced from the previously discussed endeavors suggested that terrestrial habitat in the Ray Roberts Lake area became more fragmented and complex in shape. Habitat patches became smaller but more numerous. Forested areas were most effected and conservation efforts should focus on management strategies that promote vegetation succession and forest maturation. Remotely sensed SPOT data were successfully used to predict tree basal area. There were no significant relationships found with other in situ measures or between MSS data and any vegetation measures. Remote sensing provided information suitable for large scale projects concerning landscape-level ecological issues. Rectification and classification accuracies were the …
Assessment of Changes in Aquatic Macrophyte Occurrence Following Introduction of Triploid Grass Carp in a North Texas Reservoir
The objectives of this project were to measure changes in frequency of occurrence of submerged macrophytes over the first two growing seasons following stocking with triploid grass carp at two fish per acre and to measure differences in macrophyte biomass between areas excluded from herbivory and adjacent control sites after 16 months following establishment of exclosures. The project also seeks to measure concentrations of fluridone following the herbicide treatment, and to compare two methods of aquatic vegetation sampling.
The Evolution, Applications, and Statistical Interpretations of DNA Typing in Forensic Science
This thesis examines the evolution, applications, and statistical interpretations of DNA typing as a tool in the field of forensic science as well as in our criminal justice system. The most controversial aspect of DNA typing involves the determination of how likely it is that two people share the same DNA profile. This involves the use of population genetics and databases of allelic frequencies as well as some assumptions about population structuring.
The Use of Similarity Indicies to Assess the Spatial Differences of the Phytoplankton Communities in Lake Ray Roberts, Texas
A study of the phytoplankton communities of Lake Ray Roberts was conducted at three sampling sites on two sampling dates during the summer of 1995, assessing both community structure and chlorophyll a concentrations. Eight similarity indices were then used to characterize and compare the communities of these sites. Both community similarity and chlorophyll a concentrations were evaluated using a minimum detectable difference equation to determine the amount of change that must occur in these parameters in order to be deemed statistically significant. The Bray-Curtis Index was shown to be the most adequate index evaluated, and was subsequently used in conjunction with bootstrap analysis to determine the similarity between the three sampling sites.
Evaluation of a Constructed Wetland to Reduce Toxicity from Diazinon at the Pecan Creek Wastewater Treatment Plant, Denton, TX
The City of Denton Pecan Creek Wastewater Treatment Facility has periodically failed effluent toxicity testing. A Toxicity Identification Evaluation has determined that Diazinon in the effluent is contributing to the observed toxicity. Chlorpyrifos is also implicated as a factor. The City of Denton constructed a half acre experimental wetland to remove Diazinon related toxicity. Results from spiking and microcosm experiments indicate that the wetland can reduce the Diazinon.
Short Term Effects of External Electric Fields on Electrical Activity of the Pineal Gland in Rats
The effects of short term exposure (5 minutes) to EEFs at relatively high dosages (10, 25, 39, kV/m) on the electrical activity in rat pineal glands was studied. Daytime and nighttime recordings were taken from an implanted microelectrode in the gland. The data show that (1) both the activity and frequency were enhanced when the animals were exposed to EEFs at 39 kV/m continuously and discontinuously; (2) the later condition yielded a sustained increase (36%) whereas the former a brief (10 sec) increase. This enhancement was statistically significant under both conditions (day and night). The effects observed were thought to be due to membrane alterations either in the pineal gland itself or in the neural inputs to the gland.
Biological and Toxicological Responses Resulting from Dechlorination of a Major Municipal Wastewater Treatment Plant Discharge to the Trinity River
Federal regulations such as the Clean Water Act (P.L. 92-500), and its amendments, direct the Environment Protection Agency (EPA) to implement programs to control the releases of conventional pollutants and toxics into the waterways of the United States. The EPA began requiring treatment plants to conduct toxicity tests (biomonitoring) of their effluent discharges. To control toxicity caused by chlorination of wastewater discharges, the EPA also began requiring some treatment facilities to dechlorinate their wastewater before discharging. This research was funded by the EPA to document the changes that occurred in the Trinity River from the dechlorination of the effluent from Ft. Worth's Village Creek municipal wastewater treatment plant. The study occurred over a two year period beginning in August 1990. A wide variety of biological field assessments and toxicological assays were used to measure various responses. Seven river stations, covering approximately twenty river miles, and the treatment plant effluent were assessed. Two of the river stations were upstream from the treatment plant and used as reference sites. The remaining five river stations were downstream from the treatment plant, spread out over seventeen river miles. The study evaluated the impact of chlorination prior to dechlorination, which served as a baseline. Responses determined during dechlorination were compared to the baseline data. An overall improvement in species richness and diversity was seen at those river stations which had previously been adversely impacted by chlorine. Aquatic toxicity tests, such as those required to be used by dischargers, were conducted during this study. Periodic toxicity was observed with these tests in the effluent and river samples after dechlorination was initiated. Those tests, along with in situ toxicity assays, proved to be good predictors of biological community responses.
The Response of Aquatic Insect Communities and Caged In situ Asiatic Clams (Corbicula fluminea) to Dechlorinated Municipal Effluent in the Trinity River in North Texas
Dischargers to the Trinity River in North Texas were required to dechlorinate their effluents in 1990-91. Field surveys were conducted above and below an outfall to determine the response of resident immature insects and caged in situ juvenile Asiatic clams to chlorinated and dechlorinated effluent. Within six months after dechlorination began, insect community composition and C. fluminea survival significantly improved at stations below the outfall. Significantly lower clam growth within one mile below the dechlorinated effluent indicated the presence of non-chlorine toxicants. Effects from chlorinated and dechlorinated effluent exposure were comparable between Ceriodaphnia dubia lab tests and in situ C. fluminea.
Landscape Ecological Characteristics of Habitat of the Red-cockaded Woodpecker
Geographic information systems (GIS) technology was used to analyze factors influencing habitat use by an endangered species, the red-cockaded woodpecker. The study area was the western part of the Raven Ranger District of the Sam Houston National Forest, Texas. The factors considered included both structural characteristics and spatial relationships among stands of trees in the habitat.
Comparative Toxicity Responses in Earthworms Lumbricus Terrestris and Eisenia Foetida to Cadmium Nitrate and Chlordane Using Artificial Soil and Filter Paper Exposures
This research compares LC50 and LD50 of earthworms, Lumbricus terrestris and Eisenia foetida exposed to cadmium nitrate and chlordane using 48-h contact filter paper (FP) and 14-d artificial soil (AS) protocols. Both LC50 and LD50 showed that chlordane was more toxic than cadmium in both species regardless of the exposure. The reference toxicant 2-chloroacetamide using the standardized 48-h FP exposure was used to assess the general response of the earthworm prior to toxicity experiments. A glucose test was developed as an internal standard to assess homogeneity of mixtures among both replicates and dilutions. Accuracy of dilutions is assessed by the slope of a regression line relating nominal dilution to observed internal standard concentration. Precision of replicate preparation is assessed by among replicate variance.
Changes in Heterozygosity Through Time in American Standardbred and American Saddlebred Horses (1960-1990)
Observed and expected heterozygosity (H) levels for ten electrophoretic blood marker loci and expected H for seven red blood cell (RBC) anitgen/antibody loci were examined for 20 years in American Standardbred and 30 years in American Saddlebred horses. Standardbreds were classed by gait, Trotter and Pacer, and evaluated separately in most analyses. 4,404 Trotters and 12,271 Pacers were found to have statistically highly significant losses of mean total observed H through time for the ten electrophoretic loci (P<0.005), although in Trotters the loss was more extreme (P<<0.001). Loss of H in 5984 Saddlebreds was not significant (P=0.259). Correlations of RBC expected H through time showed decreases in all three groups.
Effect of pH on the Persistence and Toxicity of Cyfluthrin to Chironomus Tentans
The effect of pH upon the aquatic toxicity of cyfluthrin was determined in 48 h static acute toxicity tests using 2nd instar Chironomus tentans larvae. Tests were conducted in both dechlorinated tap water and lake water of pH 8.0, 7.2, and 6.6. After 48 h, immobilized and dead larvae were removed and replaced with 2nd instar larvae to assess the persistence of toxicity. Midges were cultured in water adjusted to the pH values used in testing. Toxicity if cyfluthrin varied inversely with pH. An increase in the pH of tap water by 2 units resulted in a 2-decrease in toxicity. Toxicity of cyfluthrin also varied between tap and lake water of the same pH. EC50 values in lake water were approximately 2-3 times lower than those in tap water. Toxicity in the lake water and tap water at every pH tested was also significantly different when regression line elevations were compared. Natural waters amended with cyfluthrin were consistently more toxi to the chironomids than tap water of the same pH. Persistence of cyfluthrin at low pH also influenced chironomid behavior. Recovery of normal behavior generally began within 24 h at pH 8.0 At pH 6.0, recovery did not begin until one week after dosing. The persistence of cyfluthrin also varied with pH. Averaged across all concentrations, 30% of the initial dose remained in tap water (pH 8.0) after 48 h, compared with 45% (pH 7.2), and 75 % (pH 6.6).
Impacts of the Pyrethroid Insecticide Cyfluthrin on Aquatic Invertibrate Populations in Outdoor Experimental Tanks
The chemical fate and biological impacts of cyfluthrin in aquatic ecosystems were investigated using microcosms (1.9 m^3 concrete tanks) during 1989. Results were compared to a concurrent pesticide registration study using mesocosms (634.7 m^3 earthen ponds). Ten spray drift and five soil runoff simulations were conducted. Pesticide loadings were scaled by system volume, with the same experimental design in ponds and microcosms. Aqueous cyfluthrin concentrations and sediment residue values were generally higher in microcosms, while aqueous half-life was shorter in the smaller systems.
Molecular Cloning and Functional Analysis of Transposable Mercury Resistance Genes Encoded by the OCT Plasmid
Translocation of a 17.1 kilobase region of the OCT plasmid encoding mercury resistance (mer) in Pseudomonas putida was shown to occur in a recombination-deficient host with plasmid PP1 serving as a recipient replicon. The frequency of transposition in Pseudomonas was estimated at 10^3 -10 -^2, but undetectable in Escherichia soli. ' DNA comprising all of mr as well as subregions there of were cloned and subjected to DNA sequence analysis. Like other transposons, mer was found to contain inverted repeat sequences at its termini. These were similar to, but not identical to the inverted repeat structures found in the prototypical mercury resistance transposon Tn501 from E. aeruginosa.
Circulatory and Respiratory Responses to Cycle Ergometry at Different Pedal Rates
The effects of moderate workload exercise at different pedal rates on circulatory and respiratory parameters were studied. Five subjects performed seven discontinuous constant-load cycle ergometer tests of 30 minutes duration at pedal rates of 40, 50, 60, 70, 80, 90 and 100 rpm. Oxygen uptake and carbon dioxide production were determined by standard open circuit spirometry, while heart rate was recorded by electrocardiograph. The CO₂ rebreathing procedure was administered during the exercise bout in order to determine cardiac output. Blood pressure was determined for each test, and total peripheral resistance was calculated. The findings indicate that progressive increases in pedal frequency during discontinuous constant-load cycle ergometry produce progressive increases in cardiovascular, respiratory and metabolic responses and a decrease in gross exercise mechanical efficiency. The results indicate that oxygen uptake, cardiac output, heart rate, ventilation and arterial-venous oxygen difference increases curvilinearly as pedal rate increases, possibly as a result of increases in recruitment of muscle fibers and/or muscle groups. These findings suggest that circulatory and respiratory responses are due to "central command" which sets the basic efferent response pattern. However, this effector pattern is modulated by afferent input originating from the legs during progressive increases in pedal rate.
Mutagenic Potential of Tetramethylthiuram Disulfide (42-S Thiram) on the Germ Cell Stages of Drosophila melanogaster
Tetramethylthiuram disulfide (42-S Thiram), a carbamate fungicide was studied for its mutagenic potential on the germ cell stages of wild-type male Drosophila melanogaster. The mutagenicity was tested using the sex-linked recessive lethal assay (SLRL). Any lethals induced in the F2 generation were evidenced by the absence of wild-type males. Although there was an increase in mutation rates in the 42-S Thiram treated wild-type males over the control wild-type males, it was not significantly higher. According to the laboratory conditions in this preliminary study, tetramethylthiuram disulfide failed to produce mutagenic effect.
Development and Validation of a Ramping Treadmill Protocol for the On-Line Measurement of Four Aerobic Parameters
Previously, Whipp et. al. (J. Appl. Physiol.: Respirat. Environ. Exerc. Physiol. 50(1):217-221, 1981) demonstrated the feasibility of determining four parameters of aerobic function, identified as maximum oxygen uptake (μVO_2), VO_2 at anaerobic threshold (θan), the time constant for oxygen uptake kinetics (rVO_2) and work efficiency (η), using a short duration ramped bicycle ergometer exercise test. Because of the importance of being able to measure these parameters on a variety of measurement instruments a short duration ramping treadmill protocol has been developed. The ability of this protocol to determine the four aerobic parameters has been validated against conventional methods. The results of this investigation indicate that μVO_2, θan, rVO_2 and, η may be obtained from a single, short-duration ramping treadmill test.
Biological Control of the Red Imported Fire Ant by the Entomopathogenic Nematode, Steinernema Carpocapsae (Weiser)
Field trials were conducted in 1988 to evaluate the effectiveness of Steinernema (=Neoaplectana) carpocapsae (Weiser) in controlling the fire ant. Infective juveniles (IJ) of the nematode were applied as drench on 235 and 422 mounds, respectively for 2-month summer and 6-week fall evaluation periods. In comparative trials, amidinohydrazone (Amdro) was applied to 249 (summer) and 65 (fall) active mounds, with 245 (summer) and 78 (fall) untreated active as controls. Nematode treatments resulted in an average of 47% control (Abbott's formula) in summer trials and 19-88% control in the fall trials, compared with 39% and 47% control, respectively with amidinohydrazone. Active mounds treated with nematodes or amidinohydrazone had significantly fewer individuals than control mounds in summer trials.
Characterization of the OCT Plasmid-Encoded Mercury Resistance Genetic Locus in Pseudomonas putida
A 17.1 Kb genetic element encoding for mercury resistance (OCT-Hg^r) was shown to translocate from its original location on the OCT plasmid to the resistance plasmid, RPl, in Pseudomonas putida. Analysis of RPl-Hg^r recombinant plasmids revealed that insertion of mercury resistance genes into RPl could occur at a variety of sites, with all recombinants having common EcoRI restriction fragments of 9.4, 3.8, 2.3, and 1.6 Kb, derived from the insertion. Hybridization analysis suggested the existence of extensive homology between this insertion and the prototypic mercury resistance transposon, Tn501, as well as the location of a similar merA sequence. Although the overall size was shown to be quite different from Tn501, striking physical similarities are shared between these two elements.
The Reproductive Consequences of Carriers of Methylenebisacrylamide-Induced Balanced Reciprocal Translocations in Mus Musculus
N,N'-methylenebisacrylamide (MBA) was studied because of its effectiveness in inducing heritable translocations in germ cells of male mice. The health impact of translocations was studied through anatomical analysis of the progeny of semisterile translocation carriers. As expected, the semisterility of translocation carriers resulted primarily from embryonic death during periimplantation stages due to unbalanced chromosome sperm segregants. Among conceptuses that survived to mid- and late-gestation stages, there was an increased incidence of developmental anomalies including fetal death and phenotypic defects. These abnormalities are associated with unbalanced chromosome complements that allow survival to the later stages of development.
Adenylate Energy Charge Determinations of Soil Bacteria Grown in Soil Extract Medium
The adenylate energy charge values of twenty bacteria isolated from soil and cultured in a medium consisting of soil and distilled water were determined by the luciferin-luciferase bioluminescense method. The purpose of this study was to examine the growth and energy charge values of these organisms in soil extract medium, and to determine what effect the addition of glucose has on their energy charge values. Three of the organisms employed in this study showed energy charge values similar to those reported for bacteria grown in enriched media. The remainder of the isolates demonstrated low energy charge values, and scant growth in the soil medium.
Genetic Variation in a Population of the Plains Woodrat Neotoma micropus
Neotoma micropus from Jack County, Texas, were studied over a 9-month period. Loci from blood and saliva were used to determine genetic variation within the population. Deviations from Hardy-Weinberg equilibrium were found at one locus. The average temporal F over all seven loci was 0.040. Genetic structuring was subtle, fluctuated on a seasonal basis, and was due to differential migration or predation on genotypes. Heterozygotes tended to move more than homozygotes, and a greater proportion of heterozygotes were lost from the population during each season. Genetic variation was maintained in the population by immigrant individuals. This differential in dispersal of genotypes fits current models of reorganization within the genome of populations.
An Evaluation of the Short-Term Embryo-Larval and Seven-Day Larval Test Methods for Estimating Chronic Toxicity of Zinc to the Fathead Minnow (Pimephales promelas)
Chronic toxicity of zinc to Pimephales promelas was estimated by conducting replicate static and static-renewal short-term embryo-larval tests and static-renewal seven-day larval tests. The two test methods were highly reproducible. Daily renewal of test solutions had little effect on the toxicity of zinc, however, the stage of development at which exposure was initiated affected the sensitivity of the toxic endpoints measured. The most sensitive and reproducible endpoint in the embryo-larval tests was survival of viable (non-deformed) larvae and in the seven-day larval test was growth of the larvae, which was slightly more sensitive than the embryo-larval test endpoint. The estimated MATC of 0.18 and 0.15 mg/L mean total and mean soluble zinc, respectively, compared well with published results. Because of its advantages and similar sensitivity, the short-term embryo-larval test was recommended for estimating chronic toxicity.
Predicting the Site-Specific Bioavailability of Zinc Using the Indicator Species Procedure: A Case Study
National Water Quality Criteria intended to protect aquatic life and their uses from the adverse effects of pollutants may not be appropriate due to site-specific factors that alter chemical bioavailability. The Indicator Species Procedure may be used to derive site-specific criteria in order to account for differences in site-specific bioavailability. This procedure was implemented using zinc for three chemically different site (river) waters. The purpose of this study was to quantify the bioavailability of zinc in each site water and correlate results to water quality parameters and/or zinc speciation. Results demonstrated that national criteria for zinc accurately predicted the experimentally derived site-specific values within a factor of two when adjusted for water hardness. Particulate forms of zinc were shown to be biologically unavailable under conditions tested.
Aquatic Vegetation Nutrient Budgets and Sedimentation in a Southwestern Reservoir
During four growing seasons, aquatic vascular plant production and distribution were studied in Pat Mayse Lake, Texas, a 2425 hectare oligo-mesotrophic reservoir. The dominant macrophyte population was Myriophyllum spicatum L. Growth rates and regrowth rates of mechanically harvested Myriophyllum beds were found to be dissimilar. Based on estimates of watermilfoil nutrient content, there were insufficient nutrients in the entire population to alter the trophic status of this reservoir should all of the nutrients be instantaneously released. Sediments were the primary nutrient (nitrogen and phosphorus) sink. Bank erosion and solids transport from the watershed appear to contribute most of the sediments and a lake-wide mean sedimentation rate of 2.5 cm/year was estimated from sediment trap and core sample data.
Effects of Long-Term Moderate Ethanol Intake on the Stress Response in Rats
The effect of ethanol on the stress response in rats was examined. Experimental animals were given 0.25 ml of 28 percent ethanol or 0.25 ml of water orally once a day, five days a week, for a period of twelve months and were then subjected to fifteen minute cold stress. Corticosterone levels in ethanol-treated males following stress were significantly lower (22 percent) than in the sham group. Adrenal weights in sham-treated females were significantly higher (15 percent) than in the ethanol group at the end of twelve months. Mortality in sham-treated males was significantly higher (60 percent) than in ethanol-treated males. The effects observed may be due to the sedative action of ethanol on cortical centers controlling the hypothalmus.
Effects of N⁶,O²'-Dibutyryl Cyclic Adenosine 3' ,5' Monophosphate on Transformation of Rat Kidney Cells and Chick Embryo Fibroblasts by Wild-Type and Temperature-Sensitive Rous Sarcoma Virus
N^6,O^2' -Dibutyryl cyclic adenosine 3',5'-monophosphate (Bt_2cAMP) was investigated for its effects on various tissue culture cells infected with temperature-sensitive (ts) mutant, LA31 and Bratislava 77 (B77), a wild-type Rous sarcoma virus. Specifically, known parameters of transformation were investigated and a possible site of action has been tenably proposed. The drug Bt_2cAMP was found to have little effect on the transformation related properties of primary chick embryo fibroblasts (CEF) infected with either virus or normal rat kidney fibroblasts (NRK) infected with the wild-type B77-RSV. However, significant inhibition of the transforming properties in NRK infected with the ts mutant LA31 (LA31-NRK) were reported at the permissive temperature 33 degrees centigrade (33 C).
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