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Identification of Glycine as the Factor in Peptone Which Induces Pleomorphism in Azotobacter Vinelandii
The rigid peptidoglycan layer of the cell wall is responsible for maintaining the structural integrity of bacteria. Antibiotics such as penicillin exert their anti-bacterial effect by inhibiting synthesis of peptodoglycan, and enzymes such as lysozyme destroy cell integrity by hydrolyzing specific bonds in the interior of this macromolecule. Defective cells can no longer withstand the high turgor pressure within the cell because they are no longer protected by a rigid wall and tend to become fragile and spherical or irregular in shape. While all bacteria are pleomorphic under certain conditions which do not normally affect other bacteria. This is exemplified by the pleomorphic growth of Azotobacter in nutrient agar or peptone-containing medium. The purpose of this investigation was to study the nature of peptone-induced pleomorphism of Azotobacter. The first phase of study dealt with the effects of poptone on the growth and morphology of A. vinelandii. Many diverse froms were observed in peptone-containing media, but it was shown that all cell types were related to the "fungoid" family of pleomorphic cells. Although Azotobacter failed to accumulate detectable levels of cell-wall precursors in response to glycine treatment, it was shown that glycine acted only on metabolically active cells. In addition, incorporation of glycine into cell wall of Azotobacter was not required for induction of pleomorphism. Methionine and aspartic acid, and to a lesser degree alanine and isoleucine, were found to competitively inhibit glycine toxicity.
The Detection of Poliovirus in Denton Sewage by Immunofluorescence and Immunodiffusion Techniques
Several final sewage effluents from the Denton Disposal Plant were demonstrated to contain Poliovirus types II and III. Pleated encapsulated filters at pH3.5 enhanced the recovery of the Poliovirus at a higher tier in comparison with nitrocellulose filter (Millipore) and glass fiber filter of pore size 0.45u. This thesis explores problems that face us today in our quest to eliminate viral pathogens from the natural and waste water needed for human, domestic, and industrial consumption. Preliminary experiments concern the use of immunofluorescence, and immunodiffusion techniques as a means of poliovirus identification, which invariably suggests that these techniques may be useful as rapid screening procedures of water samples for presence of potentially pathogenic viruses.
Linkage Relationships of Greenbug Resistance in Barley, Hordeum Vulgare L.
The linkage relationship and arm location of the gene for greenbug resistance in the variety Will was determined by using primary trisonomics and tertiary trisomic homozygous translocations. The gene for greenbug resistance was found to be on linkage group 1 by using primary trisonomics. The gene was located on the cetromere segment of the Tl-6a translocation by using a tertiary trismoic homozygous for greenbug resistance. The data further substantiates the feasibility of using trisomics in placing genes on proper linkage groups.
Biochemical Genetics of Certain Species of the Blackbird Family Icteridae
Starch gel electrophoresis was used to compare 14 proteins encoded by 15 loci for seven species of the family Icteridae. A close genetic relationship among these species was classified into three groups. The Agelaiine group contained Agelaius phoeniceus, Sturnella magna, and S. neglecta. The Quiscaline group contained Euphagus cyanocephalus, Cassidix mexicanus, and Quiscalus quiscula. Molothrus ater, the most divergent, was placed in a separate group. Divergence times for the seven species were compared to the literature. Heterozygosity of the seven populations of the two species of Sturnella were compared to determine factors influencing their divergence. Two factors proposed were heterosis in S. neglecta and possible hybridization between S. neglect and S. magna.
The Vertical Stratification of the Macrobenthos in the Brazos River, Texas
Quantification of stream macrobenthos populations has remained a perplexing problem in rivbrine ecology, despite numerous attempts at improvement. This is in part due to well documented variations in chemical and physical parameters locally and geographically, and resultant adapted macrobenthos populations. Southwood (1968) and Hynes (1970a) have reviewed the various sampling techniques developed'for the census of lotic macrobenthos populations. Needham and Usinger (1956), Chutter (1969), and others have pointed out the difficulty in obtaining adequate numbers of samples which will yield population estimates with desired statistical confidence, and still maintain some degree of sampling economy. Needham and Usinger (1956) and Gaufin et al. (1956) mentioned the "patchy" distribution of aquatic insect populations as the primary source of this difficulty. The concept of patchy distribution in insect populations was originally discussed by Andrewartha (1961). Attempts to improve confidence through improved sampling devices and techniques have led to development of numerous types of samplers. Cummins (1962) indicated that there were almost as many samplers as there were researchers.
Effects of an Acute Bout of Near-Maximal Intensity Exercise on the Cardiac Enzymes in Human Sera
The Cardiac Profile, a pattern of serum enzyme changes seen within seventy-two hours after an AMI, is diagnostic aid for detecting occurrence of infarcts. The effects of exercise stress on the Cardiac Profile aid clinicians in avoiding diagnostic errors in patients immediately after exercise. Five male volunteers ran from six to ten miles. Serum enzyme levels were monitored serially three days before and five days after stress. Enzyme activity was determined spectrophotometrically and electrophoretically. Significant increases in total CPK and LDH were seen. An LDH 'one-two flip' occurred eight hours after exercise. No MB-CPK was found following the run.
Comparative Chemistry of Thermally Stressed North Lake and Its Water Source, Elm Fork Trinity River
To better understand abiotic dynamics in Southern reservoirs receiving heated effluents, water was analyzed before and after impoundment in 330 ha North Lake. Macronutrients, metals, and chlorinated hydrocarbons were measured. Concentrations of nutrients and metals in sediments were quantified in this 2 yr study. River water prior to impoundment contained 16 times more total phosphorus, and supported 23 times more Selenastrum capricornutum cells in an algal assay than reservoir water. The reservoir has essentially no drainage and since evaporation is high, the concentrations of many dissolved solids have increased since the reservoir was filled in 1958. North Lake is now phosphorus limited. Apparently altered chemical equilibria have caused precipitation or adsorption of phosphorus with calcium and iron.
X-irradiation Effects on the Action Potentials of Frog Sciatic Nerves Inhibited by Carbon Monoxide and Ouabain
The response of frog sciatic nerve action potentials to x-irradiation and metabolic (carbon monoxide) or transport (ouabain) inhibition was determined in an attempt to further identify the nature of radiation insult to nervous tissue. Carbon monoxide, ouabain (2 X 10-5 M), and nitrogen anoxia were shown to produce a near linear decline in action potential amplitude. The carbon monoxide and nitrogen inhibitions of activity were reversible in air; the carbon monoxide inhibition was light reversible. Ouabain inhibition was partially reversible by soaking the nerve in aerated Ringer's. Application of 120 kv x-rays (75 Kr at 4.9 Kr/min) to nerves during the linear decline in spike amplitude brought about a marked enhancement (146%) of inhibition by 99% CO/l% 02, nitrogen (136%), and ouabain (265%). All bhanges were shown to be statistically significant by a regression analysis. However, x-irradiation did not appear to alter the air reversibility of carbon monoxide and nitrogen inhibitions nor the reversibility in Ringerts of the ouabain inhibition. Additionally x-irradiation completely blocked light reversal of 98% CO/2% 02 inhibition and produced a decline in activity. A possible interpretation of these results is a compensation for radiation action at this dosage requiring metabolism and ion pump activity.
Evolutionary Genetics of Three Semispecies of Wood Rats--Neotoma Albigula, Neotoma Micropus, and Neotoma Floridana
Electrophoretic variation in 18 proteins encoded by 20 autosomal loci was used to compare the genetic relationships of 19 natural populations representing three species of the subgenus Neotoma. Of the 20 loci examined nine were monomorphic and fixed for the same allele in all populations. No more than seven loci were polymorphic within a single population. Genetic variability was expressed as the proportion of loci heterozygous in the average individual of a population. Heterozygosity in the three species of Neotoma studied averaged 0.078, a value within the range reported for other rodents. Although the levels of heterozygosity seen in Neotoma could not readily be explained, the variation may be attributed to ecological factors. The three species of Neotoma were compared on the basis of genetic similarity and found to form a close taxonomic unit, probably semispecies. Divergence times were obtained for the three species and found to compare well with divergence times obtained from fossil data. In general, the three species have diverged within the last 112,000 years during the Wisconsin glacial period.
The Stoneflies (Plecoptera) of Texas
An illustrated key to the adults and known nymphs of Texas Plecoptera is provided. Species accounts, including geographic distribution within Texas, and biological notes are given. Of the twenty-seven species of Plecoptera known from Texas, six are new state records. TWo species new to science, Isoperla jewetti and Isoperla coushatta are described. Taeniopteryx starki Stewart and Szczytko, Zealeuctra arnoldi Ricker and Ross, and Zealeuctra hitei Ricker and Ross are endemic to the Edwards Plateau area of Texas. Two species, Mesocapnia frisoni (Baumam and Gaufin) and Isoperla jewetti New Species are western in origin. The remaining nineteen species (excluding Anacroneuria) are typically eastern species.
The Effect of Ethanol on Cardiac Activity and Brain Respiration in Chick Embryos
This study concerned the effect of ethanol on cardiac activity and brain respiration in chick embryos. Ethanol dosages tested ranged from 1.0 mg to 4.0 mg/g weight. Each experiment lasted at least 150 minutes. Cardiac activity in terms of total waveform energies was integrated and printed out for plotting and analysis. The embryonic heart rates were simultaneously determined from physical graph tracings. The embryonic brain respiration was measured using a differential microrespirometer. The effect of ethanol on cardiac activity was one of slight (10 to 13), but statistically significant (p<.05) rate depression. The brain slices exhibited a marked, immediate, and irreversible decrease (39 to 89%) in oxygen consumption at both ethanol dosages. The data indicated that chick embryonic brain tissues were more susceptible to alcohol effects than cardiac tissue. Therefore, the mental abnormalities seen in the offspring of alcoholic mothers may be more cerebral in nature than cardiovascular.
Pelagic Phytoplankton and Physicochemical Correlates for Lake Texoma
An analysis and correlation of phytoplankton communities with physicochemical data from 3 sites in Lake Texoma was conducted to supplement time-series data. Water and phytoplankton were sampled monthly, March, 1976-February, 1977. Simple correlations were run between all physicochemical parameters and phytoplankton standing crop from the 3 sites. Multiple linear regression analyses were used to develop equations predictive of phytoplankton standing crop and chloride concentration. Minerals leached from marine sediments in the Red River chennel contribute to formation of a, halocline which seals the anoxic hypolimnion from the reservoir surface in midsummer. Conductivity decreased west to east, 2980-1800. pmhos/cm. Maximum mean annual phytoplankton standing crop in Red River arm was 36 percent greater than midlake. Eutrophication was evident.
Cigarette Residues Affect Steroidogenesis in Cultured Y-1 Mouse Adrenal Tumor Cells
This study (1) quantitatively compared steroid production in cultured Y-l mouse adrenal tumor cells exposed to Camel and Carlton-smoke derived residues, and (2) localized the effects in the cell. Basal steroid production was increased by Camel residues but not by Carlton, while ACTH stimulation was interfered with by both residues. Camel basal stimulation was comparable to that of cAMP, and was abolished by Cytochalasin D. The stimulation was also comparable to that of cholera toxin, which activates adenyl cyclase. Results indicate that residue components dissolve in the membrane stimulating adenyl cyclase at a point similar to or before that utilized by cholera toxin for its stimulating effect.
Effects of Turbidity on Gilling Rates and Oxygen Consumption on Green Sunfish, Lepomis cyanellus
Laboratory studies conducted at 5, 15, 25, and 35 C measured changes in gilling rates and oxygen consumption of green sunfish in response to exposure to bentonite clay suspensions. The tests indicate that gilling rates are not affected by bentonite clay suspensions below 2125 FTU at 5 C, 1012 FTU at 15 C, and 898 FTU at 25 C. At turbidity levels exceeding 1012 FTU at 15 C and 898 at 25 C, gilling rates increased 50-70%. Tests were inconclusive at 35 C. Oxygen consumption rates were found to be unaffected by turbid suspensions below 3500 FTU at all four temperatures. Evidence suggests that increased gilling rates under highly turbid conditions are a means of compensating for reduced respiratory efficiency and a strategy for maintaining a constant oxygen uptake. Evidence indicates that the cost of increased gilling rates is probably met by a reduction in activity.
The Distribution of Pathogenic Pseudomonas aeruginosa in Sewage
The purpose of this study was to extend our understanding of the ecological relationships of P. aeruginosa by investigating the differences or similarities between the strains of this organism found in sewage and those found as pathogens in human infections. This research was approached by comparing the serological types of P. aeruginosa isolated from sewage contaminated waters in Argentina (South America) to those isolated from sewage contaminated waters in Texas. They were typed with sera obtained using P. aeruginosa isolated from human infections. The data obtained revealed that bacteria isolated from sewage in Texas and from soil and water in Argentina are antigenically similar to those isolated from human infections.
Genic Differentiation and Evolution in the Ground Squirrel Subgenus Ictidomys (Spermophilus)
The genetic structure of 26 natural populations of three species (S. tridecemlineatus, S. mexicanus, and S. spilosoma) of the Ictidomys subgenus of ground squirrels was analyzed using chromosomal and electrophoretic techniques. Chromosomal variation was not observed in S. mexicanus, and only slight karyotypic variation was found in the other two species. Chromosomal evidence indicated hybridization between S. tridecemlineatus and S. mexicanus, placing these species within the classical definition of semispecies. Analysis of electrophoretic variation at 29 genetic loci indicated close genetic relationships between these species. Evolution in Ictidomys appears to be linked with Pleistocene events, and speciation appears to have occurred within the last 155,000 years.
The Life History of the Mayfly Isonychia sicca (Walsh) (Ephemeroptera--Siphlonuridae) in an Intermittent Stream in North Central Texas
The life history of Isonychia sicca (Walsh) was elucidated from samples collected at Clear Creek from Oct. 1976-Jun. 1978, and Elm Fork of the Trinity River from Sept. 1977-Jun. 1978, Denton County, Texas. Adaptations for existence in an intermittent stream were of primary concern. Eggs are capable of diapausing through hot, dry summers and cold, wet or dry winters. Diapause is broken in the fall after rehydration and/or in the spring. I. sicca is usually bivoltine during a Sept.-Jul. wet period. Observations from Elm Fork indicate that emergence continues to Oct. if the stream remains permanent. Considerable overlap occurs between overwintering, spring, and summer populations.
Dielectric Properties of Azotobacter vinelandii in a Microwave Field
A resonant frequency cavity was used to determine the dielectric properties of various preparations of Azotobacter vinelandii ATTC 12837. It was found that the bacteria investigated did interact with microwave radiation in the absence of free water. The data presented here indicate that bacteria demonstrate frequency specific dielectric properties. The techniques employed in these experiments may also be used to determine microwave spectra of other species of bacteria in different physiological stages.
Production and Energy Metabolism in Three Benthic Insect Populations in a Small North Central Texas Pond
Annual energy budgets of dominant benthic macro-invertebrates were examined during November 1973 to October 1974 from the benthos of a small pond ecosystem in north-central Texas. Estimates of annual secondary production (Hynes and Coleman 1968) were Procladius s. (Diptera, Chironimidae), 2.4 g m^-2 y^-1 (13 kcal m^-2 y^-1 ); Tendipes decorus (Diptera, Chironomidae), 6.0 g m^-2 y^-1 (40 kcal m^-2 y^-1 ); Brachycercus sp. (Ephemeroptera, Caenidae), 1.9 g m^-2 y^-1 (11 kcal m^-2 y^-1). Energy metabolism was measured in the laboratory at six seasonally encountered temperatures (5, 10, 15, 20, 25, and 30 C) on an acclimatization basis, and then extrapolated to the field. Estimates of annual energy metabolism are Procladius sp., 5.0 kcal m^-2 y^-1 ; Tendipes decorus, 17.2 kcal m^-2 y^-1 ; Brachycercus sp. 40.0 kcal m^-2 y^-1.
Effect of Operant Behavior on the Metabolism of 5-Hydroxytryptamine
The role of operant behavior in the metabolism of brain 5-hydroxytryptamine (5-HT) turnover was investigated. Two and one-half hours following the administration of 150 mg/kg of para-chlorophenylalanine (PCPA), a specific inhibitor of tryptophan hydroxylase, levels of 5-HT were compared in sedentary and performing rats. Whole brain levels of serotonin were reduced in both responding and sedentary animals; however, differences between these groups were not statistically significant. The drug induced decrease in 5-HT levels was accompanied by a significant decrease in session responding. The degree of suppressed responding could be correlated with the level of 5-HT following PCPA, suggesting that the metabolism of serotonin is in part modulated by the rate of responding as maintained by the operant schedule.
A Study of the Water-Soluble Antigens from Virulent and Attenuated Biotypes of Brucella abortus
Through chemical analysis and ion exchange chromatography of watersoluble antigens, this investigation supports the view that the majority of differences between the biotypes are quantitative. It was also found that strains demonstrate distinct, qualitative differences when compared to the attenuated strain 19 by immunodiffusion and thin-layer polyacrylamide gel, isoelectric focusing. These differences include the presence of antigens on virulent strains that are absent on strain 19. In addition, one antigen absent on strain 19, was found common to each virulent biotype. Finally, the results from immunodiffusion experiments, employing adsorbed and non-adsorbed immune globulins, indicate that at least some water-soluble antigens are exposed on the cell surface and that their distribution among the biotypes varies.
The Relation Between Carbon Assimilation and Biomass Dynamics in a Phytoplankton Community
Production dynamics in the phytoplankton community of a mesotrophic Texas reservoir were measured weekly over a four month period using 14C incubation and ATP assay methodologies. Assimilation values of 14C ranged from 0.2 to 45 ug C 1 hr1 - . Significant positive and occasionally negative changes in biomass (i.e. viable organic carbon) were observed in short term (4 hr) in situ incubations juxtapo-sitioned with the 14C experiments; viable organic carbon production, estimated with the ATP assay, ranged from -25 to +50 ug C 1l1hr1. Carbon assimilation and biomass changes did not correlate in either short term (4-5 hr.) or over the study period (6 months). However, weekly biomass trends were predicted by relative positive or negative biomass changes in the short term incubations. Biomass measurements gave a more sensitive insight into production dynamics in the phytoplankton community than did carbon assimilation measurements.
The Passage of Sodium-24 and Rubidium-86 Across the Blood-Brain Barrier System of Canines at Low Body Temperatures
To evaluate the blood-brain barrier system in the pathogenesis of an irreversible hypothermic state in dogs, concentrations of 2 4 Na and 86Rb were measured at body temperatures ranging from 37 0 C to 160 C. A suppression of transport of sodium was demonstrated, followed by an increase as the temperature was lowered. The concentration of rubidium ion increased in concentration as the temperature fell. These data indicate there may be a temperature threshold below which the blood-brain barrier system fails to maintain the internal environment of the central nervous system. The intimate relationship of several brain stem nuclei with the cerebro-spinal fluid indicates they may be at risk during profound cooling.
Food Habits of Stoneflies (Plecoptera) in the Gunnison and Dolores Rivers, Colorado
Gut contents of 2,500 stonefly nymphs, comprising 10 species, from the Gunnison and Dolores Rivers, Colorado were examined from Dec., 1974-Oct., 1975. Perlidae species were carnivorous feeding primarily on chironomids, mayflies and caddisflies. Seasonal patterns of ingestion and preference varied among species and predator sizes and between rivers. Early instar polyphagous species utilized detritus in the fall, eventually shifting to carnivorous habits as they grew through winter-spring. Pteronarcids fed predominantly on detritus. Dietary overlap of predators was greatest in the Gunnison River, with subtle mechanisms such as prey species and size selectivity, temporal succession and seasonal shifts to detritus-plant material in some, providing reduction of competition. A more complete partitioning of prey resources was evident in the Dolores River.
Trophic Structure and Energy Flow in a Texas Pond
Annual energy flow and mean annual biocontent of eighteen compartments were determined for a 0.94 ha north central Texas pond ecosystem. Annual primary production was 7,780 kcal m^-2 yr^-2, and community production-to-respiration ratio was 1.49. One-third of annual primary production accumulated on the substrate as silt and sedimentation. Community production, production-respiration ratio, and biocontents of all compartments except aquatic insects were large in summer, small in winter. Biocontents of four trophic levels in the pond were all of the same order of magnitude, approximately 50 kcal m^-2. Suspended and benthic organic material forprimary consumers and terrestrial insects for tertiary consumers were substantial allochthanous energy imports into the pond system.
Drumming Behavior of Selected Nearctic Stoneflies (Plecoptera)
Drumming was recorded for 11 of 13 Nearctic stonefly species, representing 4 families. Both male and female signals were obtained from 5 species, and were either 2-way or 3-way communications. Signals were species-specific; those of males and females varied from 3-39 and 1-14 beats/ signal, respectively. Duration of male signals varied from 105-8,016 ms; those of females, except Perlinella drymo (1 beat), varied from 402-1318 ms. Signals among related taxa showed greatest similarities. Duration of male signals of Perlinella drymo became progressively shorter at each of 4 temperatures from 7-29 0C. Females of Perlinella drymo would only repeatedly answer male signals recorded at near their own temperature, and would not repeatedly answer recorded male signals of 8 other species.
DNA-DNA Hybridization of Methane Oxidizing Bacteria
Bacteria classified in the family Methylomonadaceae must derive their carbon from one-carbon compounds. They are characterized by the possession of internal membranes of two types. Type I membranes are layered and fill the middle of the cells while type II membranes form concentric layers around the periphery of the cells. Also, there are two metabolic pathways by which the methylobacteria assimilate one-carbon compounds. Further evidence of this dichotomy was sought by DNA-DNA saturation hybridization of DNAs from both types of methylobacteria. Very low DNA-DNA homology was seen between types I and II or within the types. It was not possible, therefore, to correlate the degree of genetic relatedness with either the nature of the internal membranes or the pathway of carbon assimilation.
Histochemical Characterization of Lymphocytes in Preleukemic and Leukemic AKR Mice
The AKR strain of mice have a genetic trait for spontaneous development of lymphocytic leukemia. In this study, leukemic mice were found to have significantly larger (p<0.01) thymuses and spleens than preleukemic mice. The enlarged leukemic tissues were densely packed with a light staining cell, with a hollow-appearing nucleus. Tissues from preleukemic mice were observed to be infiltrated with a smaller, darker-staining lymphocyte. Fluorescent antibody staining was done on preleukemic and leukemic tissues, using three antisera against murine lymphocyte theta antigen, and an antiserum against murine IgG. Significantly brighter fluorescence, (p <0.05) with theta-specific antisera, was found in leukemic thymuses,spleens, and kidneys than in the same preleukemic tissues. Leukemic tissues had significantly brighter fluorescence (p <0.05) than preleukemic tissues with IgG antiserum.
Nymphs of the Stonefly (Plecoptera) Genus Taeniopteryx of North America
Nymphs of the 9 Nearctic Taeniopteryx species were reared and studied, 1976-78. Two morphologically allied groupings, the Taeniopteryx burksi-maura, and T. litalonicera- starki complexes corresponded with adult complexes. A key separating 7 species, based primarily upon pigment patterns and abdominal setal arrangements, was constructed. Taeniopteryx lita and T. starki were indistinguishable; T. burksi was separated from T. maurawhen no developing femoral spur was present. This study was based upon 839 nymphs. Mouthparts were not species-diagnostic. Detailed habitus illustrations were made for 6 species. Egg SEM study revealed that 3 species were 1.2-1.4 mm diameter, with a highly sculptured chorion, generally resembling a Maclura fruit; micropyle were scattered. Taeniopteryx lita, lonicera, starki and ugola nymphs were described for the first time.
The Evolutionary Genetics of Campostoma anomalum and Campostoma oligolepis
Electrophoretic variation in 12 proteins encoded by 16 loci was analyzed to compare the genetic relationships of 18 natural populations representing two species of stoneroller minnows, Campostoma anomalum and C. oligolepis. Ten of the loci were monomorphic and fixed for the same allele in all populations of both species. One locus, Mdh-2, was found to separate both species. Mean heterozygosity for both species was 0.072. Estimates of levels of inbreeding indicated this phenomenon is operating in C. anomalum to structure the populations genetically. Mean genic identity (I) between the two species was high, 0.887, indicating the taxa are closely related. Nevertheless, data accumulated point to the conclusion that the two species maintain electrophoretic variation in 12 proteins encoded by 16 loci was analyzed to compare the genetic relationships of 18 natural populations representing two species of stoneroller minnows, Campostoma anomalum and C. oligolepis. Ten of the loci were monomorphic and fixed for the same allele in all populations of both species. One locus, Mdh-2, was found to separate both species. Mean heterozygosity for both species was 0.072. Estimates of levels of inbreeding indicated this phenomenon is operating in C. anomalum to structure the populations genetically. Mean genic identity (I) between the two species was high, 0.887, indicating the taxa are closely related. Nevertheless, data accumulated point to the conclusion that the two species maintain their genetic integrity throughout their ranges. their genetic integrity throughout their ranges.
Distribution, Abundance, and Food Habits of Larval Fish in a Cooling Reservoir
Analysis of larval fish collected at four stations in a 330-ha cooling reservoir indicated Dorosoma spp. were most numerous at all stations, followed by Lepomis spp. and Percina Macrolepida. Largest numbers and greatest diversity of larval fish were found at the station least affected by thermal effluent; the mid-lake station provided the smallest numbers and least diversity. The two warmwater stations were intermediate, with similar numbers and diversity. Diversity and abundance of zooplankton between stations were similar to those of fish. The most abundant zooplankter (Bosmina) was generally selected against by Dorosoma, Lepomis and Micropterus spp. larvae except when the larvae were quite small ((10mm). Cyclopoid copepods were most often selected by all larvae.
An Assay Method for Determining Extra-Cellular Lipases from Pseudomonas aeruginosa
The applicability of an isotopically labelled assay system to determine the lipase production in Pseudomonas aeruginosa was evaluated. Supernatant from cultures of Pseudomonas aeruginosa grown in a medium containing olive oil was incubated with a substrate containing labelled trioleate. Fatty acids were isolated by means of a liquid-liquid partition system. Enzyme activity was determined by measuring the amounts of free fatty acid by liquid scintillation counting. Findings indicate that the isotopicallylabelled, liquid-liquid partitioning assay is reliable, sensitive and adaptable to rapid assay conditions. It was also determined that different strains of Pseudomonas aeruginosa produce varying amounts of lipase. Partial purification of supernatant by gel filtration produced two protein peaks showing enzymatic activity.
Effects of Ribavirin on Normal Rat Kidney Cells and Chicken Embryo Fibroblasts Infected with Rous Sarcoma Virus
Ribavirin, a synthetic nucleoside, was found to inhibit the replication of Rous sarcoma viruses (RSV) and subsequent cell transformation in chick embryo fibroblasts (CEF). It also blocked the transformation of normal rat kidney (NRK) cells infected with temperature-sensitive mutants of RSV. The action of Ribavirin was found to be reversible as removal of the drug from the NRK cells reversed the effects on cell transformation. Ribavirin appears to have a static effect on cell growth of both NRK and CEF cells. In addition, guanosine, xanthosine and inosine altered the effect of Ribavirin on cell growth.
Thermal Selection at an Enzyme Locus in Populations of the Red Shiner, Notropis lutrensis, Receiving Hypolimnion Effluents from a Reservoir
Genetic variation was examined at 19 loci encoding enzymatic and general proteins Notropis lutrensis from the Brazos River in Texas. The thermal regime of the Brazos River below Possum Kingdom Reservoir is altered due to the release of water from the hypolimnion. Summer water temperatures fluctuate as much as 7^oC. Levels of heterozygosity at the malate dehydrogenase-2 locus were correlated with the degree of water temperature fluctuation at each locality. The isozymes from three homozygous patterns of supernatant malate dehydrogenase (Mdh-l, Mdh-2) exhibited different activities at different experimental temperatures.
Immunocompetence in the AKR Mouse
A model for the study of the relationship of immunity to cancer is found in AKR mice which harbor Gross virus. This genetically transmitted virus is present in a latent form for months before it spontaneously induces leukemia. Many investigators have demonstrated near normal humoral responses, but abnormal cellular immunity in the preleukemic animal. With increasing age, pathology of the disease is expressed, reflecting diminished immunity. In this study, the ontogeny of humoral antibodies of AKR/J and SWR/J mice was assayed by microagglutination techniques in response to thymus-independent, thymus-dependent, and solubilized antigens. Simultaneous injections of thymusdependent and -independent antigens provided data suggesting an impaired humoral response in the AKR mouse.
Use of ATP as a Planktonic Biomass Indicator in Reservoir Limnology
A series of laboratory experiments and a field investigation were conducted to closely define the application of the ATP assay and ATP as a planktonic biomass estimator for routine use in reservoir limnology. The laboratory experiments verified the published range of C:ATP ratios (i.e. 250:1) as a conversion factor for ATP to biomass in cultured selected genera of freshwater algae, except for the species of blue-green algae examined. The field investigation conducted at Moss Reservoir included organic carbon measurements with ATP biomass in size classes on a depth basis. The ATP biomass varied seasonally and with depth; the best significant mtltiple correlation was between organic carbon and the smallest size class (.45 to 10 um) and total net plankton biomass (.45 to 165 um). Daily monitoring of biomass in size classes demonstrated the sensitivity of the technique.
Serological Relationships of Azotobacter Species
In order to clarify the taxonomic problems which exist among the group azotobacter by serological method, 24 cultures of Azotobacter including 9 species were tested as antigens in Ouchterlony plates against 3 different antisera obtained from rabbits immunized with A. chroococcum ACl6, A. macrocytogenes St.M. and A. vinelandii 12837, respectively. The results showed more cross reactivity in intraspecies reactions than in interspecies ones and indicated the presence of a common immunogenic determinant. This work presents the results which suggest the effecient classification of Azotobacter by Ouchterlony comparisons of corresponding antigens in different species.
Effect of Amino Acids on Growth and Cartenogenesis in Corynebacterium Species Strain 7E1C
Studies were evaluated on the effects of known growth factors on the growth and carotenogenesis of Corynebacterium species strain 7ElC. The complex medium, Tryptic Soy Broth,was found to stimulate growth and production of more pigment in the light and in the dark than did a mineral salts-glucose medium. A complete amino acid mixture added to LSG enhanced carotenogenesis in the dark in Corynebacterium 7ElC, while B-vitamins retarded carotenogenesis. No absolute requirement for one or more amino acids was found,indicating a multiple amino acid requirement. The fewest amino acids found to stimulate carotenogenesis in the dark were a combination of those in the Serine and Histidine families which include serine, glycine, cysteine, and histidine.
Hematological Parameters of the Bluegill, Lepomis machrochirus (Rafinesque), Including Effects of Turbidity, Chloramines, and Flexibacter columnaris
Normal ranges of values for hematological parameters of bluegill gathered seasonally from three lakes were determined. Sexual, seasonal, and inter-lake variations were found. Effects of 2-wk exposure to turbidity on blood parameters included an increase in rbc size and a decrease in small lymphocytes. Effects of 3-hr exposure were increases in rbc count, hemoglobin, and pH and decreases in PG2 and large lymphocytes. The effects of 0.44 and 0.88 ppm chloramines were an increase in blood pH, a decrease in MEV, and severe spastic reactions resulting in loss of equilibrium or death in 90% of the fish. Effects of Flexibacter columnaris included an increase in transformed lymphocytes and a decrease in small lymphocytes.
Isolation and Characterization of Malic Enzyme from Ascaris suum
A procedure for the isolation of malic enzyme from muscle tissue of the roundworm Ascaris suum is described. The fractionation method yields relatively large quantities of the enzyme,with a specific activity of fifteen moles of malate converted to pyruvate and carbon dioxide per min per mg at 25º. Homogeneity was established with analytical ultracentrifugation, zone electrophoresis, isoelectric focusing, and rechromatography. The molecular weight of the enzyme was 250,000, and it is dissociated under several conditions into four identical monomers of 64,000 daltons. The enzyme exists as a single electrophoretic form and prefers manganous and NAD over other cations and NADP. Ammonium sulfate competes with manganous for the active site and titration with DTNB yields eight thiol groups per mole. Titration of the first four thiol groups is accompanied by a complete loss in enzyme activity. Equilibrium dialysis, product inhibition, and initial velocity studies suggest a rapid-equilibrium random sequential mechanism for the Ascaris suum malic enzyme. The presence of 1.3 binding sites per subunits was determined for L-ma late. Antisera prepared against A. suum malic enzyme reacted to a small extent with the NAD malic enzymes from two free-living nematodes, Panarellus redivivus and Turbatrix aceti. A correlation coefficient of 0.911 was obtained upon comparing the amino acid composition of A. suum and E. coli malic enzymes. Some sequence homology is predicted between these malic enzymes. The physiological interpretation favors the binding of malate initially, with the subsequent addition of NAD to the enzyme.
Biochemical Systematics of the Genus Sophora
Three unusual amino acids, y-amino-n-butyric acid, pipecolic acid, and 4-hydroxypipecolic acid, and an uncommon dipeptide, y-glutamyltyrosine, have been isolated and characterized from the seeds of members of the genus Sophora. Structural proof of these compounds was carried out by paper chromatography, thin-layer chromatography, column chromatography on amino acid analyzer, infrared, nuclear magnetic resonance, mass spectrometry, and C, H, N analysis. The presence and absence of these compounds was used as a criterion for the classification of 23 species of the genus Sophora. A phylogenetic classification which seems to follow the morphological taxonomy of this genus was carried out on the basis of seeds that contained pipecolic acid, those which did not contain pipecolic acid, and plants which contained both pipecolic acid and 4-hydroxypipecolic acids. Another chemical classification was also introduced based on the presence and absence of y-amino-n-butyric acid and y-glutamyltyrosine.
Trophogenic Ecology of Selected Southwestern Reservoirs
Three north central Texas reservoirs (Grapevine Reservoir, Lake Ray Hubbard, Lewisville Reservoir) were investigated in order to characterize the nutritional ecology, phytoplankton community structure, and primary productivity within the trophogenic zones of each. Emphasis was placed on elucidating the relative influences of the major nutrients (C, N, P, Fe, Si) and various other physico-chemical parameters on phytoplankton community biomass, structure, and productivity. Extensive physical, chemical, and biological analyses consisting of approximately eighty parameter measurements were made on each of twenty-three integrated water samples (surface to three meters) taken from the three reservoirs during the period July, 1971 to June, 1972. New methods which were employed included a high-intensity ultraviolet photocombustion procedure for the determination of total nitrogen, phosphorus, and iron, and an in vitro oxygen method for estimating community metabolism. General chemical and physical regimes are described for the three reservoirs, and various interrelationships discussed. Phytoplankton communities are delineated on bases of species composition, volume, diversity, pigments, and metabolism data. The more salient biotic and physico-chemical interrelationships are examined within the context of the limiting nutrient controversy.
Evolutionary Genetics of Certain Mice of the Peromyscus boylii Species Group
The genetic structure of 49 natural populations of four species (P. attwateri, P. boylii, P. pectoralis, and P. polius) of the Peromyscus boylii species group was analyzed through application of chromosomal and electrophoretic techniques. Chromosomal variation within and among populations of the boylii species group was analyzed from 178 specimens. Electrophoretic techniques were utilized for the demonstration of variation in enzymes and other proteins encoded by structural loci and applied to the study of the evolution of the boylii species group by estimation of levels of genetic heterozygosity within populations, estimation of degree of genetic similarity between conspecific populations and between species, and determination of patterns of geographic variation in allelic frequencies and levels of heterozygosity. Six distinct chromosomal patterns were observed among the populations of the four species of the boylii species group. All specimens had a diploid number of 48 and the major difference in chromosomal morphology was in the number of pairs of large to medium biarmed autosomes. Little or no chromosomal variation was observed in three species (attwateri, pectoralis and polius), but considerable chromosomal variation occurred among populations of P. boylii. Generally, the chromosomal variation in P. bylii was between allopatric populations, with each chromosomal pattern limited to a recognized subspecies. Polymorphism was observed in two populations. The polymorphism observed in P. polius was the result of pericentric inversion involving the smallest pair of metacentric autosomes. The polymorphism observed in P. bolii cileus was interpreted at the result of gene flow between P. boylii rowleyi and P. boylii spicilegus. In addition to chromosomal evidence, analysis of electrophoretic data demonstrated and suggested effective gene flow between the chromosomal forms of P. boylii. Electrophoretically demonstrable variation was analyzed in 11 proteins encoded by 17 autosomal loci. Of the 17 structural loci, 11 were polymorphic in one or more …
Biochemical Genetics of the Pocket Gopher Genus Geomys, and its Phylogenetic Implications
Electrophoretic techniques were utilized for the demonstration of variation in 22 proteins from 24 natural populations of four species ( G. bursarius, G. pinetis, G. arenarius and G. personatus ) of the Geomys complex of pocket gophers. Of the 24 structural loci , 19 were considered to be polymorphic. Five of the six esterases contributed greatested to the polymorphism while non-esterase proteins generally showed low values. In the GeoMys complex of pocket gophers in this study,selection appeared to be the most important influence on genetic structure with some evidence of random drift in two of the four species. Populations of G. arenarius and G. personatus had the highest average interspecific genetic similarities to G. bursarius and . pinetis was the most divergent. Biochemical evidence supports the phylogeny of Geomys based on morphological and fossil data.
Population Genetics, Karyology, and Morphology of Certain Species of the Peromyscus Truei Group
The systematic relationship of two species of the Peromyscus truei group (P. truei and P. difficilis) was analysed through the application of starch gel electrophoresis, numerical taxonomy, and chromosomal techiques. Of 20 loci examined, 11 were monomorphic in all populations, two exhibited variation in only two populations, and seven loci were polymorphic in two or more populations. The mean number of polymorphic loci per population was 0.186, the mean number of polymorphic loci per individual was 0.024, and the proportion of loci heterozygous per individual was 2.4%. Chromosomal forms of P. truei, P. t. gentilis (FN 54) and P. t. truei (FN 62), and P. difficilis, P. d. petricola (FN 56) and P. d. nasutus (FN 58), were consistent for their karyotypes throughout their geographic ranges. No chromosomal hybrids were detected. Numerical analysis of morphological characters and similarity values based on allelic frequencies utilizing Roger's coefficient (S) demonstrated a distinct seperation of karyotypic forms of P. truei (S = 0.902) and P. difficilis (S = 0.924) and were below the mean value of S for conspecifics (S = 0.950). All data indicates that each chromosomal form of the P. truei group examined represents four distinct species. The oldest available name for chromosomal forms of P. true with a fundamental number of 62 is Peromyscus truei Shufeldt (1885) and the oldest available name for chromosomal forms with a fundamental number of 54 is Peromyscus gratus Merriam (1898). The oldest available name for chromosomal forms of P. difficils with a fundamental number of 58 is PeromYSCus nasutus Allen (1891) and the oldest available name for chromosomal forms with a fundamental number of 56 is Peromyscus difficilis Allen (1891).
Ecological Energetics of the Dobson Fly, Corydalus Cornutus
Rates and energies of consumption (C), egestion (F), assimilation (A), respiration (R), growth (Pg), production of exuviae (Pev), and production of egg masses (Pr) and associated efficiencies, and the effects of seasonal temperature, weight and metamorphic stage upon these factors were examined for a typical individual and cohort of Corydalus cornutus (L.) from a stream in North-Central Texas (330 23'N, 97*5'W). Dobson flies are apparently univoltine in the study area, with 11 larval instars. Emergence, oviposition and hatching occur from late May to August. The typical dobson fly hatches in mid-June, grows rapidly until November, and resumes rapid growth in March, reaching full adult size prior to leaving the stream to pupate in early June. Adult females must feed to provide energy to yolk eggs, produce egg-mass coverings and continue somatic maintenance during their week of reproductive endeavors. Metabolic compensation enables larval dobson flies to maintain preferred and fairly constant rates of R during winter (201-451 pl g-1 h~1; 5-15 C) and summer (985-1173 pl g- h1; 20-30 C); with a seasonal acclimatization change point between 15-20 C. Reduction of rates of R through undercompensation during the winter when food is scarce and through partial compensation at high temperatures during the summer conserves energy which is allocated to P, resulting in high ratios of P/R (1.94) and P/A (66%) for the individual larva and, to a lesser degree, for the cohort (P/R = 1.07, P/A = 52.3%, P/B = 9.96). Rates of C, F, A and R, but not assimilation efficiency, were influenced by temperature and size. The energy budget for a typical dobson fly during the 47 wk as a larva was: C = 4167, A = 3442, F = 725, Pg = 2075, Pev = 198, and R = 1169. Ova respired 0.107 cal wk-1, prepupae 357 cal …
A Study of the Interaction of Co-Insult Treatments with Methylmercuric Chloride and X-Irradiation and Demonstration of a Peroxide Induced Protective Mechanism
The initial purpose of this work was to investigate the interaction of methylmercuric chloride (MMC) and X-irradiation given as a co-insult upon the rat blood-brain barrier (BBB). The indicators used to determine BBB alterations were mortality and the in vivo tissue uptake of radioactive sulfate administered as 3 5S-sodium sulfate. The results of the interaction studies indicated a neutralization of effects when MMC and X-irradiation were given together. X-irradiation as a single insult generally caused an increase in sulfate uptake by the brain regions monitored, whereas MC treatment generally resulted in decreased sulfate uptake. The neutralization patterns following co-insult treatments were somewhat varied in the different brain regions, exhibiting cancellation of effects in some cases and overriding by one insult in other eases. From the data obtained by this work and in the literature, it is hypothesized that the P-L organelle system of the perivascular glia serves as a trap for MMC, preventing MMC from reaching the neurons. The system appears to proliferate in response to increased peroxides in the body fluids, thereby increasing tolerance to larger doses of MMC.
Microbiological Treatment of Wastewater from a Wood-Preserving Plant
This research investigates interacting biological, chemical, and physical factors affecting the efficiency of microbiological wastewater treatment at the W. J. Smith Wood- Preserving Company in Denison, Texas. The treatment process consisted of collecting exhaust boiler water containing unidentified boiler treatment compounds, steam condensate contaminated with preservatives and wood extracts, plant process waters, and rainfall runoff from plant grounds. With a 5-minute residence time, wastewater was passed over 2 oxidation towers in series, each containing approximately 47,000 square feet of surface area. Suspended solids were removed from the wastewater before discharge. Various amino acids such as serine, aspartate, cysteine, phenylalanine, alanine, proline, glycine, histidine, and tyrosine significantly stimulated phenol degradation in the laboratory. The plant wastewater contained approximately 0.1 mg/l of several of the stimulatory amino acids. It was assumed that these concentrations provided maximal stimulation in the field situation. The plant wastewater also contained sufficient nitrogen to permit the organisms to degrade up to 100 mg phenol/1 of water examined. Amino acids in the wastewater probably serve as a source of microbial nutrition. Toxicity of the wastewater to fish was not caused by the presence of phenol, phenol degradation products, or traces of pentachlorophenol. The wastewater was rendered non-toxic by diluting with between 4 to 9 volumes of stream water. Toxicity could also be removed by chemical coagulation followed by activated carbon adsorption. As a result of biological treatment, the plant now discharges the treated wastewater into the municipal sewage treatment facility.
In Vivo and In Vitro Transformations of Mouse Tissues from a Murine Lymphosarcoma
The problem with which this investigation is concerned is that of determining the nature of events leading to the change. of normal cells into malignant cells. The design of the study is multi-phasic: (A) to establish the presence or absence of an oncogenic virion, (B) to demonstrate by use of the electron microscopy any ultracellular alteration in malignant or transformed tissues, (C) to investigate the nature of the transforming agent in the murine lymphosarcoma, and (D) to employ various methods to demonstrate cellular transformations in vivo and in vitro. It is concluded that the transforming and tumorinducing agent in this' investigation was not a virion, but an infectious ribonucleic acid genome or a segment of a viral genome which had become integrated into the genome of the mouse cells. The vision has lost its ability to form a protein coat; therefore it is not demonstrable as a virion. But the ribonucleic acid is able to infect other cells and transform them from normal to neoplastic tissues.
Aspects of the Thermal Ecology of Largemouth Bass (Micropterus salmoides) in North Central Texas
The coefficient of body temperature change (K) ranged from -0.53 to -0.072 for bass weighing 73-1440 g. The double log regression of K on weight was similar to that reported for other poikilotherms (slope = -0.57; R = 0.93). Fingerling bass were eurythermal, being capable of surviving instantaneous temperature changes over a 20 C range at acclimation temperatures of 15, 25 and 30 C and over a 15 C range at acclimation temperatures of 20 and 35 C. Preferred temperatures for adult bass measured in the laboratory ranged from 27-32 C with no relationship to day or night. The overall mean preferred temperature was 29 C. The laboratory determined preferred temperatures were supported by limited field determined body temperatures taken in a vertical temperature gradient near the discharge of a power plant effluent. Routine metabolic rates of bass from a heated reservoir and a nearby hatchery were similar from 10-30 C in summer and winter. The weight exponent (0.77) and Q^gS (1*6-2.9) were similar to those published for more northern bass populations; however, the Texas bass had lower metabolic rates than those published for the northern populations. Bass exposed to rapid temperature increase (0.2 C/min) from 25-30 C increased their metabolic rate by 53% but showed no detectable increase in opercular rate. Bass warmed from 30-35 C and 30-33 C increased their metabolic rate by 140%, and their opercular rates increased to over 100 beats per minute before death.
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