This study identifies the biogeographic factors that structure small mammal communities on mountains of the Colorado Plateau and adjacent regions. Forty six isolated ranges were characterized across a 5-state study area encompassing the Colorado Plateau, including the central high plateaus of Utah and the Basin and Range Province (i.e. the Great Basin and mountains of Arizona and New Mexico). Presence/absence data of 25 montane mammal species were used to explore the interactions between historical and ecological processes affecting local and regional diversity patterns. Multivariate analyses, such as non-metric dimensional scaling, were used to explore factors which influence community composition. Results of these analyses revealed the Colorado River as a significant biogeographic barrier that affects montane mammal community structure. MtDNA cytochrome b sequence variation was analyzed among populations of the long-tailed vole, Microtus longicaudus, sampled from five interior ranges of the Colorado Plateau- Abajo, LaSal, Henry, and Chuska Mts., and Boulder Mountain of the Aquarius Plateau-and analyzed using traditional phylogenetic approaches (parsimony and likelihood) as well as nested clade analysis. Results support previous documentation of a major east-west phylogeographic break occurring between populations southeast of the Colorado River (eastern Arizona, Colorado, Wyoming and New Mexico) and all other western populations, which …

Food waste in the waste stream is becoming an important aspect of integrated waste management systems. Current efforts are composting and animal feeding. However, these food waste disposal practices rely on slow thermodynamic processes of composting or finding farmers with domestic animals capable of consuming the food wastes. Bioconversion, a potential alternative, is a waste management practice that converts food waste to insect larval biomass and organic residue. This project uses a native and common non-pest insect in Texas, the black soldier fly, which processes large quantities of food wastes, as well as animal wastes and sewage in its larval stage. The goal of this research is to facilitate the identification and development of the practical parameters of bioconversion methods at a large cafeteria. Three major factors were selected to evaluate the practicality of a bioconversion system: (1) the biological constraints on the species; (2) the economic costs and benefits for the local community; (3) the perception of and interaction between the public and management agencies with respect to the bioconversion process. Results indicate that bioconversion is feasible on all levels. Larvae tolerate and consume food waste as well as used cooking grease, reducing the overall waste volume by 30-70% …

Cowpea, Vigna unguiculata (L.) Walp is an excellent source of protein, vitamins and minerals and a major food crop many parts of Africa. Optimal production levels are hampered by insect pests and diseases. Biotechnological techniques such as tissue culture and genetic engineering can aid in the development of varieties with resistance to insect pests and diseases. The objective of this study was to investigate conditions necessary for the development of a reproducible tissue culture system that can be applied to regenerate transformed cells from culture. The in vitro manipulation of cowpea using Murashige and Skoog (MS) medium, auxins and cytokinins resulted in the formation of callus and rhizogenesis. Calli that were formed were separated into six classes based on color and texture. Yellowish friable callus, yellowish compact, soft yellowish callus and green and white were composed of largely vacuolated cells and were non-regenerative. Friable green callus was the most prevalent callus type and could form of roots in some hormone combinations. Green spots were formed on hard compact green callus. The green spots became nodular, forming root primordia and ultimately giving rise to roots. None of the six calli types gave rise to the formation of shoots. Embryogenic callus was …

N-Acylethanolamines (NAEs) are endogenous lipid metabolites that occur in a variety of dry seeds, and their levels decline rapidly during the first few hours of imbibition (Chapman et al., 1999, Plant Physiol., 120:1157-1164). Biochemical studies supported the existence of an NAE amidohydrolase activity in seeds and seedlings, and efforts were directed toward identification of DNA sequences encoding this enzyme. Mammalian tissues metabolize NAEs via an amidase enzyme designated fatty acid amide hydrolase (FAAH). Based on the characteristic amidase signature sequence in mammalian FAAH, a candidate Arabidopsis cDNA was identified and isolated by reverse transcriptase-PCR. The Arabidopsis cDNA was expressed in E. coli and the recombinant protein indeed hydrolyzed a range of NAEs to free fatty acids and ethanolamine. Kinetic parameters for the recombinant protein were consistent with those properties of the rat FAAH, supporting identification of this Arabidopsis cDNA as a FAAH homologue. Two T-DNA insertional mutant lines with disruptions in the Arabidopsis NAE amidohydrolase gene (At5g64440) were identified. The homozygous mutant seedlings were more sensitive than the wild type to exogenously applied NAE 12:0. Transgenic seedlings overexpressing the NAE amidohydrolase enzyme showed noticeably greater tolerance to NAE 12:0 than wild type seedlings. These results together provide evidence in vitro …

A number of studies have shown that sensory deprivation is associated with selective decreases in GABA, GAD, and GABA receptors, in deprived areas of visual and somatosensory cortex. Those studies focused on layer 4, a recipient of direct thalamocortical sensory input. However, supragranular layers 2/3 have been recently identified as a major locus of functional plasticity in sensory deprivation and long-term potentiation. To examine whether GABAA receptors in layers 2/3 are affected by sensory deprivation, rats had mystacial vibrissae in middle row C or rows ABDE trimmed for 6 weeks beginning in early adulthood. Layers 2/3 above the deprived and adjacent whisker barrels were located in tangential sections, using patterns of radial blood vessels as fiducial marks. In deprived whisker barrel columns, [3H]muscimol binding to GABAA receptors decreased by 12.8% ± 1.2 (P &lt; 0.001) in layers 2/3 and 11.4% ± 1.2 (P<0.001) in layer 4. Altered levels of GABAA α1 subunit (Fritschy et al., 1994) were indicated by reduced optical density of immunostaining, both in deprived layers 2/3 (6.4% ± 0.7; P&lt; 0.001) and in layer 4 (3.4% ± 1.0; P &lt; 0.005). Interestingly, Nissl staining density also decreased in deprived layers 2/3 (12.7% ± 1.8 P &lt; 0.001) …

The toxicity of methanol, atrazine, and copper to Pseudokirchneriella subcapitata (Korshikov) Hindák historically referred to as Selenastrum capricornutum Printz were determined following 96 hrs growth in a modified Goram's growth media. Methanol and atrazine inhibited fluorescence readings in the cultures by 50% (IC50) at concentrations of 2% and 82 µg/l respectively. These toxicity values compared favorably to other published reports. The IC50 for copper was 160 µg/l which is substantially higher than reported values. This is understandable because of the high chelating capacity of Goram's media. The use of stereologically derived relative volume in the chloroplasts, mitochondria, lipid bodies, phosphate bodies, and nucleus was investigated to determine if it could be used as a sensitive endpoint in toxicity tests. The volume fractions for the chloroplasts and mitochondria were normally distributed in control cells while the nuclei, phosphate bodies, and lipid bodies were not. The chloroplasts were the most dominate organelle occupying a mean relative volume of 46% and mitochondria occupied a mean relative volume of 3%. The nucleus and phosphate bodies occupied a median relative volume of 7% and 2% respectively. The lipid bodies were rare in section profile and no meaningful median relative volume could be calculated. Up to …

From August 1996 through September 1997 eleven fixed stations were sampled monthly in January, March , April , July, August, September, and November and fortnightly in May and June for the purposes of establishing baseline conditions present in Lake Texoma as related to U.S. Army Corps of Engineers chloride control activities in the upper Wichita River, Texas. Five reservoir zones were identified a priori using historical chloride concentration data and include the Red River Zone (RRZ), Red River Transition Zone (RRTZ), Main Lake Body (MLB), Washita River Transition Zone (WRTZ), and Washita River Zone (WRZ) in order of decreasing chloride concentration. The existence of the WRTZ is not supported here, however the Big Mineral Arm in the RRTZ was observed to be highly independent of the mixing patterns observed in the RRTZ and was treated post priori separately from the RRTZ. Spatial and temporal comparisons between reservoir zones were performed on seventeen (17) physicochemical parameters from each of the eleven sampling stations and phytoplankton count data from one sampling station within each reservoir zone and physicochemical parameters were observed to exhibit a fixed spatial gradient. Strong density gradients throughout the reservoir were observed to occur in conjunction with vertical stratification …

The Rio Gavilan watershed, located in Mexico 's northern Sierra Madre Occidental , has significance in conservation history. Upon visiting the remote, largely un­developed watershed during two hunting trips in the 1930s, renowned conservationist Aldo Leopold thought it was the best picture of land health he had seen. His main indicators of healthy land were slow water runoff rates regulating erosion and historical predator-prey relationships. The visits confirmed Leopold's concept of land health, inspired many of his essays, and helped shape his land ethic. Leopold proposed the area as a control site to research healthy land throughout North America . The proposal never went forward and the area has since been more intensively logged and grazed. This dissertation research used extensive literature review, archives, oral histories, citizen surveys, and rapid assessment of forest, rangeland, riparian, and socioeconomic health to assess impacts of past cultures and update the area's land health status. Projects that could restore land health, such as linked eco-tourism, forest density reduction, and rotational grazing, were assessed for feasibility. Recent critiques of Leopold's land ethic were also reviewed. Results indicate most pre-1940s impacts were light, current land health status is moderate, and local interest exists in restoring land …

Burkholderia cepacia is a common soil bacterium of significance in agriculture and bioremediation. B. cepacia is also an opportunistic pathogen of humans causing highly communicable pulmonary infections in cystic fibrosis and immunocompromized patients. The pyrB gene encoding ATCase was cloned and ATCase was purified by the glutathione S-transferase gene fusion system. The ATCase in B. cepacia has been previously classified as a class A enzyme by Bethell and Jones. ATCase activity gels showed that B. cepacia contained a holoenzyme pyrBC complex of 550 kDa comprised of 47 kDa pyrB and 45 kDa pyrC subunits. In the course of purifying the enzyme, trimeric subunits of 140 kDa and 120 kDa were observed as well as a unique proteolysis of the enzyme. The 47 kDa ATCase subunits were cleaved to 40 kDa proteins, which still demonstrated high activity as trimers. The proteolysis site is between Ser74 and Val75 residues. To confirm this, we converted the Ser74 residue to an Ala and to an Arg by site-directed mutagenesis. After this primary sequence changed, the proteolysis of ATCase was not observed. To further investigate the characteristics of B. cepacia pyrB gene, a pyrB knock-out (pyrB-) was constructed by in vitro mutagenesis. In the assay, …

Cyanide utilization in Pseudomonas fluorescens NCIMB 11764 occurs via oxidative conversion to carbon dioxide and ammonia, the latter satisfying the nitrogen requirement. Substrate attack is initiated by an enzyme referred to as cyanide oxygenase (CNO), previously shown to require components in both high (H) (>30 kDa) and low (L) (<10 kDa) molecular weight cell fractions. In this study, tetrahydrobiopterin (H4biopterin) was identified as a cofactor in fraction L, thus making CNO appear as a pterin- dependent hydroxylase. CNO was purified 150-fold (specific activity 0.9 U/mg) and quantitatively converted cyanide to formate and ammonia as reaction products. When coupled with formate dehydrogenase, the complete enzymatic system for cyanide oxidation to carbon dioxide and ammonia was reconstituted. CNO was found to be an aggregate of known enzymes that included NADH oxidase (Nox), NADH peroxidase (Npx), cyanide dihydratase (CynD) and carbonic anhydrase (CA). A complex multi-step reaction mechanism is proposed in which Nox generates hydrogen peroxide which in turn is utilized by Npx to catalyze the oxygenation of cyanide to formamide accompanied by the consumption of one and two molar equivalents of oxygen and NADH, respectively. The further hydrolysis of formamide to ammonia and formate is thought to be mediated by CynD. The …

This dissertation research explored the capabilities of neuronal networks grown on substrate integrated microelectrode arrays in vitro to be applied to toxicological research and lipid signaling. Chapter 1 details the effects of chlorine on neuronal network spontaneous electrical activity and pharmacological sensitivity. This study demonstrates that neuronal networks can maintain baseline spontaneous activity, and respond normally to pharmacological manipulations in the present of three times the chlorine present in drinking water. The findings suggest that neuronal networks may be used as biological sensors to monitor the quality of water and the presence of novel toxicants that cannot be detected by conventional sensors. Chapter 2 details the neuromodulatory effects of N-acylethanolamides (NAEs) on the spontaneous electrical activity of neuronal networks. NAEs are a group of lipids that can mimic the effects of marijuana and can be derived from a variety of plant sources including soy lecithin. The most prominent NAEs in soy lecithin, palmitoylethanolamide (PEA) and linoleoylethanolamide (LEA), were tested individually and were found to significantly inhibit neuronal spiking and bursting activity. These effects were potentiated by a mixture of NAEs as found in a HPLC enriched fraction from soy lecithin. Cannabinoid receptor-1 (CB1-R) antagonists and other cannabinoid pathway modulators indicated …

TNT (2,4,6-trinitrotoluene) is a persistent contaminant at many military installations and poses a threat to aquatic ecosystems. Data from environmental fate and toxicity studies with TNT revealed that sediment toxicity test procedures required modification to accurately assess sediment TNT toxicity. Key modifications included aging TNT-spiked sediments 8-14 d, basing lethal dose on measured sediment concentrations of the molar sum of TNT and its main nitroaromatic (NA) transformation products (SNA), basing sublethal dose on average sediment SNA concentrations obtained from integration of sediment SNA transformation models, avoiding overlying water exchanges, and minimizing toxicity test durations. Solid phase microextraction fibers (SPMEs) were investigated as a biomimetic chemical measure of toxicity and bioavailability. Both organism and SPME concentrations provided measures of lethal dose independent of exposure scenario (TNT-spiked sediment or TNT-spiked water) for Tubifex tubifex. Among all benthic organisms tested (Chironomus tentans, Ceriodaphnia dubia, T. tubifex) and matrixes, median lethal dose (LC50) estimates based on SPME and organism concentrations ranged from 12.6 to 55.3 mmol SNA/ml polyacrylate and 83.4 to 172.3 nmol SNA/g tissue, ww, respectively. For Tubifex, LC50s (95% CI) based on SNA concentrations in sediment and SPMEs were 223 (209-238) nmol SNA/g, dw and 27.8 (26.0-29.8) mmol SNA/ml, respectively. Reproductive effects …