In order to gain a better understanding of this microorganismand its role in human pathogenesis, a physical map of Moraxella catarrhalis type strain ATCC25238 was constructed using pulsed field gel electrophoresis (PFGE) in combination with Southern hybridization techniques. Restriction endonucleases Not I, Rsr II, and Sma I were used to digest the chromosomal DNA. An overlapping circular map was generated by cross-hybridization of isolated radiolabeled fragments of Moraxella catarrhalis genomic DNA to dried PFGE gels. The number and location of the 16S and 23S ribosomal RNA genes were determined by digestion with l-Ceul enzyme and by Southern hybridization. Virulence-associated genes, the gene for β-lactamase, and housekeeping genes were also placed onto the physical map.

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.

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 problem of using a state adopted textbook written to apply to a large body of students with varying interests and needs was overcome by using a detailed syllabus that arranged course content in a meaningful sequence that appealed to student interest. The outlined syllabus prepared a two semester life science curriculum to be used by the teacher to guide lesson planning. Both semesters were divided into three units each. Materials included in the syllabus were given to actual student groups in real classroom settings. Since hands on learning was an important part of classroom instruction, two laboratory sections were included in the appendices to be used with the syllabus.

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.

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.

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.

Principal objectives of my research were to: (1) report for the first time that coelomocytes are able to reduce NBT dye and confirm the presence of lysozyme-like activity in earthworm; (2) develop a standard methodology for determination of NBT reduction and lysozyme-like activity in earthworms; (3) compare NBT reduction and lysozyme-like activity in earthworms with those of murine and human cells and fluids; and (4) demonstrate the sensitivity of earthworm NBT reduction and lysozyme-like activity as the assays using matrics in refuse-derived fuel fly ash (RDFF) and CuSO4.

The effects of cannabinoid agonists on spontaneous neuronal network activity were characterized in murine spinal cord and auditory cortical cultures with multichannel extracellular recording using photoetched electrode arrays. Different cultures responded reproducibly with global decreases of spiking and bursting to anandamide and methanandamide, but each agonist showed unique minor effects on network activity. The two tissues responded in a tissue-specific manner. Spontaneous activity in spinal tissue was terminated by 1 μM anandamide and 6.1 μM methanandamide. Cortical activity ceased at 3.5 μM and 2.8 μM respectively. Irreversible cessation of activity was observed beyond 8 μM for both tissues and test substances. Palmitoylethanolamide, demonstrated that CB2 receptors were not present or not responsive. However, the data strongly suggested the presence of CB1 receptors.

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.

This study investigated whether acute myocardial ischemia of the anterior left ventricular wall induced an increase in cardiac sympathetic efferent nerve activity and thereby affected regional myocardial blood flow and contractile function.

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. …

The research presented herein provides information on coelomocyte (leukocyte) collection, function and immunotoxicity from polychlorinated biphenyls (PCB) in the earthworm Lumbricus terrestris. Research was undertaken as part of an overall goal to develop a well-documented and scientifically valid non-mammalian surrogate immunoassay with the earthworm Lumbricus terrestris to assess immunotoxic potential of xenobiotics. The principal objectives were to: (1) Develop an extrusion model for analyzing immunotoxicity of xenobiotics; (2) determine if coelomocytes can be collected repeatedly without obvious harm to the earthworm or change in immune response of the coelomocytes harvested and (3) validate the response sensitivity profiles of a panel of biomarkers {differential and total cell counts, erythrocyte rosette (ER) and secretory rosette (SR) formation with, and phagocytosis of antigenic rabbit red blood cells} in earthworms after acute exposure to a known mammalian and L. terrestris immunotoxin, the PCB Aroclor 1254.

Members of a guild of cursorial spiders (Pardosa spp. and Alopecosa spp.) spatially segregated among five discrete habitats, from a streamside cobble habitat grading laterally along a successional gradient to the leaf litter zone of a transition or climax riparian forest. Seasonal activity peaked in midsummer for all guild members. Spiders were active diurnally earlier in the streamside habitats, and levels of activity were positively correlated with light intensity. Guild members Pardosa tristis and Pardosa uncata were most different in habitat selection and periods of diurnal activity. Males and females of all guild species differed in their distribution among habitats and over months of collection. Measures of guild species diversity and evenness were variable between habitats, and were largely influenced by the relatively high abundance of one or two guild species, particularly in the streamside habitats.

Concern over sediment toxicity has increased the need for toxicity test information with organisms that inhabit sediments. Oligochaetes are exposed to toxicants through feeding and direct body contact with aquatic sediments. Chronic testing with oligochaetes has historically focused on tubificids with test lengths of one year or more to encompass several generations. Most naidid oligochaetes have generation times of three to seven days and could provide chronic information in a matter of weeks. The cosmopolitan distributed naidid, Pristina leidyi, was evaluated for use as a toxicity test organism. Results of research conducted includes culture methods, effects of temperature on reproduction, growth rates in a reference sediment, acute toxicity tests, and chronic toxicity tests.

A unique, previously undescribed Gram-negative bacterium was isolated from several soils in Texas and extensively characterized in this study. The cells measured 1-2 by 4-6 μm. The distinguishing characteristic of the bacterium is the extraordinary capsular material which surrounds the cells. The new isolates are aerobic, mesophilic, non motile and have the ability to utilize a variety of organic compounds as the sole source of carbon and energy. The organism grows optimally at 30° C and the optimal pH lies between 7.0-8.0. The isolates produce catalase but oxidase is not produced. They do not produce indole or hydrogen sulfide. The organism can hydrolyze gelatin and Tween 80 but not starch, esculin and casein. The major cellular fatty acid is anteiso 15:0. The guanine and cytosine content is 58-62 mole%. The organism's taxonomic position was further established by specific gene probes, 16S rRNA homology, DNA homology and "ribotyping." These data showed that it was most closely related to members of the genus Paenibacillus, although somewhat divergent from other species classified in this genus. After careful evaluation of the results obtained during this study, it is proposed that this unique bacterium be named Paenibacillus velasolus sp. nov.

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.

Culture health of Ceriodaphnia dubia was evaluated for organisms cultured using all combinations of three foods and three waters. Criteria used to assess health of cultures included adult and neonate weights, time required to produce first broods, neonate production, adult survival, and resistance to hexavalent chromium. Diet/water combinations which produced the most neonates were not found to produce adults which were more resistant to chromium than those which produced fewer neonates. Of those evaluated, a diet of Selenastrum capricornutum and a yeast-trout chow-cereal leaf mixture was best for culturing and testing Ceriodaphnia. The best synthetic water tested was a mixture of nine parts reconstituted hard water and one part bottled mineral water.

Ancient peoples did not distinguish between philosophy, religion, and science. Scientific truth did not exist apart from divine truth. Any new idea, finding, or theory was assimilated into a monolithic mythological structure. This is one of the causes of the underestimation of ancient science: it is always packaged in a myth - the method of preserving information in an oral culture. The mythological medium allowed the preservation and dissemination of hard-won, empirical, scientific knowledge through generations of preliterate peoples. The context for mythological memorization, or simply tradition, needed to be easily and naturally acquired. The ideal context was the anthropomorphic context, the ontogenic context. This is the Grand Allegory - the anthropomorphization of information. Biomyths are essentially biological texts allegorized in esoteric language.

Concurrent outdoor aquatic 1950 L microcosm and 0.04 ha mesocosm experiments with bluegill sunfish evaluated the ecological impact of cyfluthrin. Cyfluthrin effects were not observed on mesocosm bluegill; a slight decrease in growth was observed in the microcosm bluegill. Otolith weight to length relationships between bluegill size-classes from microcosms, local streams, and a fish hatchery revealed no differences. Our results indicated bluegill predation impacts were slight on benthic invertebrates. Extensive predation on emerging insects was observed. Microcosm bluegill impacts on zooplankton populations followed expected predation effects, resulting in larger populations of smaller taxa. Bluegill functioned as "keystone" predators for microcosm taxa and improved taxa richness for benthic colonizing invertebrates and zooplankton.

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 …

Acclimation temperature had a significant effect on the lethality of copper to fathead minnows in replicated 96-hour lethality tests. Lowest median lethal copper concentrations (LC50) were recorded at 12 and 22°C, with LC50s at 5 and 32°C at least 140 μg/1 higher. This research found LC50 copper concentrations in the 300 to 500 μg/1 range and a polynomial relationship between LC50s and acclimation temperature. Following a 24-hour exposure to three sublethal concentrations of copper, critical thermal maxima (CTMax) were tested in minnows acclimated to four temperatures. Sublethal exposure to copper significantly decreased the CTMaxs relative to controls at three of the four temperatures. Control CTMaxs ranged from 28.6 to 40.4°C and increased 0.46°C for each 1°C increase in acclimation temperature.

The purpose of this project is to produce a complete and thorough notebook that may be used to supplement laboratory coursework. Its intent is to be used primarily by the students to aid them in understanding background information and the proper laboratory procedures involved in various types of experiments. The laboratory notebook is a summation of all the experiments and procedures used in the six-credit hour Advanced Microbial and Molecular Biology (BIOL 5160) course offered during the summer semester at the University of North Texas. This class is a team taught effort by Professors O'Donovan and Kunz. The course is constructed as an intensive practice exercise to teach the student about gene mutations, biosynthetic pathways, preparation and analysis of plasmid DNA, and many other topics included in the notebook.

Pseudomonas fluorescens does not appear to regulate the enzymes of de novo pyrimidine biosynthesis at the level of gene expression. Little or no apparent repression of pyr gene expression is observed upon addition of exogenous pyrimidines to the growth medium. The Escherichia coli pyrBI genes for aspartate transcarbamoylase (ATCase) were sized down and cloned into the broad host range plasmid, pKT230. Upon introduction into a P.fluorescenspyrB mutant strain, ATCase showed repression in response to exogenously fed pyrimidine compounds. Thus, it was possible to bring about changes in pyrimidine nucleotide pool levels and in transcriptional regulation of gene expression at the same time.