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Biological Sciences.

Papers presented at the 83nd Annual Meeting

Troy University

Troy, Alabama

March 15-18, 2006

LARGE WOODY DEBRIS AND ITS INFLUENCE ON MACRO-INVERTEBRATE ASSEMBLAGES IN SOUTHEASTERN COASTAL PLAIN STREAMS, USA. Sameer Bharrarai and Michael W. Mullen, Dept. of Environmental and Biological Sciences, Troy Univ., AL 36082

Large woody debris (LWD) has important structural and functional roles in streams, but little research on the function of LWD has been performed in the southeastern coastal plains. This study surveyed LWD in 35 stream reaches in the Choctawhatchee River watershed in SE Alabama. The study reaches were a mix of urban, forested and mixed stream drainages. LWD characteristics and functions were quantified. Pool and sediment characteristics and drainage areas of each reach and used with already existing macroinvertebrate data. Statistical analyses of LWD, pool, sediment, drainage and macroinvertebrate data were used to examine the relationship among LWD characteristics and functions, pool, drainage and sediment characteristics, and biological variables. There were significant differences in LWD number and volume among the three site categories. These differences apparently resulted in a higher frequency of pools, sediment storage sites and woody debris storage areas, and more ecologically important, heterogeneous habitat in forested streams that contained higher number and volume of LWD. Large woody debris positively influenced macroinvertebrate assemblages and partially abated the negative effect of fine sediment on macroinvertebrates, as indicated by significant positive correlations among LWD, pools and biological variables and an apparent increase in the complexity of in-stream habitat.

DNA TESTING OF E. COLI FORMS IN SURFACE WATERS USING BOX AND REP PCR TECHNIQUES. Brian S. Burns, Dept. of Biology, Judson College, Marion, AL 36756. Stephanie M. Hood, Judson College, Marion, AL 36756.

PCR and agarose gel electrophoresis techniques were used to differentiate between human and bovine fecal point source pollution. BOX AIR (5'-CTACGGCAAGGCGACGCTGACG-3') or primers REP 1R (5'-IIIICGICGICATCIGGC-3') and REP 2I (5'-ICGICTTATCIGGCCTAC-3') were used to generate DNA fragments. Escherichia coli isolates were obtained from cows in the lower Cahaba water shed. Human samples were obtained from Judson College students. Escherichia coli isolates were also obtained from Dry Creek in the lower Cahaba water shed.

MITOCHONDRIAL DNA VARIATION IN WALLEYE AND SAUGER. Neil Billington, Dept. Biol. Environ. Sci., Troy Univ., Troy, AL 36082.

Walleye (Sander vitreus) and sauger (S. canadensis) are carnivorous percid fish species that are extensively managed by fisheries agencies because they are highly valued by anglers. Information on the genetic structure of these two species will assist in management decisions. Restriction fragment length polymorphisms of whole-molecule mitochondrial DNA (mtDNA) was examined for 135 sauger from 17 populations, and more than 1000 walleye from 100 populations, spanning the natural range of both species. Little genetic variation was observed in sauger, only five haplotypes were identified and little geographic structure was seen. These data are consistent with sauger using a single (Mississippian) glacial refugium during the Pleistocene. Walleye mtDNA haplotypes can be separated into five main geographic groups, three of which can be related to post-Pleistocene recolonization events from Atlantic, Mississippian and Missourian glacial refugia. A forth walleye group consists of a unique and genetically divergent haplotype found in the Mobile drainage. The fifth group consists of a mixture of haplotypes of glacial origin from fish recently stocked into these areas on top of a number of relic haplotypes found in the New River, VA, the Rockcastle River, KY, and the Upper Ohio River systems. Several haplotypes show localized distributions that serve as markers for stock identification. These results will prove useful for making genetic conservation decisions regarding the management of these two species.

KIDNEY STONES IN AN ATLANTIC BOTTLENOSE DOLPHIN. Gerald T. Regan, Marterra Foundation, 4000 Dauphin Street, Mobile, AL 36608-1791.

I had participated in the Alabama portion of the Southeastern United States Marine Mammal Stranding Network for 17 years, performing the necropsies of close to 400 dead-stranded specimens of the Atlantic bottlenose dolphin, Tursiops truncatus, without observing any kidney stones. On April 15, 2004, however, a 242 cm long freshly dead female specimen appeared on the beach on the south side of Dauphin Island in Mobile County, Alabama, that had stones. The necropsy was performed the next day on property generously made available by the Marine Resources Division of the Department of Conservation and Natural Resources. There were five corpora albicantia on the two ovaries combined, which is evidence for the age of the specimen, in the vicinity of 20 years. Using the protocol of cutting each kidney into cross-sectional slices about the thickness of a slice of sandwich bread, I was not surprised at finding a typically healthy appearance for the right kidney. A little over half way down the left kidney, the knife began to scrape on some hard objects. Further inspection showed that the objects were kidney stones apparently filling both minor and major calyces. The largest was over 3 cm long, and the rest were of ever-decreasing lengths and diameters. Daniel F. Cowan of the University of Texas Medical Branch oversaw the chemical analysis that showed the stones to be constituted of ammonium acid urate.

FLUORESCENCE STUDY OF SUNSCREENS IN HETEROGENEOUS NANOSPHERE MEDIA. Rajagopal Krishnan and Thomas Nordlund, Department of Physics, University of Alabama at Birmingham.

Sunscreens applied to skin are primarily observed to be located in a complex environment of outer epidermal layer called stratum corneum. Studying basic optical properties of sunscreens which are supposed to act like an optical filter is difficult in a realistic environment like heterogeneous epidermis. Hence, UV absorption and transmission properties of sunscreens are often studied in solvent which does not mimic the skin, or they are studied in complex artificial skin systems which are difficult to handle and expensive to perform. Studying the fluorescence emission of sunscreens in heterogeneous nanosphere environment and comparing the result with emission of sunscreens in skin cell and excised skin, we concluded that the polystyrene spheres in buffer can serve as mimic for skin, by presenting polar/apolar environment observed in living tissues. We will present the fluorescence emission results of octyl salicylate, octyl methoxy cinnamate and padimateO and our proposed method to obtain their approximate protection factor by using the fluorescence spectroscopy and polystyrene nanospheres. Acknowledgements: Supported in part by a grants from NCI (CA94327), NIH grants R01 CA86172 (CAE), P30 AR050948 (UAB Skin Diseases Research Center) and from VA grant 18-103-02 (CAE) and from the University of Alabama at Birmingham (Preparing Future Faculty award and GAPF graduate fellowship, support of R. K.).

RUDBECKIA AURICULATA (PERDUE) KRAL PARASITIZED BY ASPHONDYLIA RUDBECKIAECONSPICUA OSTEN SACKEN (DIPTERA: CECIDOMYIIDAE) IN ALABAMA. Alvin Diamond, Dept. of Biological and Environmental Sciences, Troy University, AL 36082

Rudbeckia auriculata (Perdue) Kral is a rare wetland associated species endemic to the southeastern United States. This is the first report of parasitism by Asphondylia rudbeckiaeconspicua Osten Sacken (Diptera:Cecidomyiidae) on R. auriculata in Alabama. No information regarding parasites has previously been reported for this species in the literature or in status surveys for the U. S. Fish and Wildlife Service. Infected populations contained over one thousand flowering stems and the parasite has not been observed at smaller populations.

EFFECTS OF SYMBIOTIC SOIL MICROORGANISMS OF DIFFERENT ECOTONES IN THE INVASION OF KUDZU IN SOUTH ALABAMA. Patricia Kinney and Dr. S. Greipsson, Dept. Biol. and Env. Sci., Troy Univ. Troy, AL 36082. Dr. H. El-Mayas, Dept. Biol. Georgia State Univ. Atlanta, GA 30302.

Invasion of exotic plants is a great threat to biodiversity and ecosystem function. Kudzu [Pueraria montana (Lour.) Merr.] is a notoriously invasive, exotic legume that is commonly found in South Alabama. Kudzu damages ecosystems by smothering native vegetation as it limits the space, water, sunlight, and nutrients available to native plants. We investigated the possible role of the symbiotic soil microorganisms rhizobia and arbuscular mycorrhizal fungi (AMF) in the invasion process. Soil samples were collected from four ecotones in Pike County, Alabama: (1) where native plant species are abundant, (2) where P. montana dominates, (3) at the forest edge, and (4) from stockpiled soil. Kudzu seeds were germinated on sterilized 0.75% water agar, and seedlings were placed in pots (n=3). Plants were harvested after eight weeks, and growth attributes were measured. Roots were examined for nodules, and rhizobia was extracted and analyzed by REP-PCR. Roots were stained, and AMF root colonization was estimated. Kudzu plants grown in soil collected from native plants had significantly highest numbers of root nodules, leaf numbers, plant length, biomass, and dry mass than those of plants in other treatments. The results suggest that native symbiotic soil microorganisms facilitate the invasion of kudzu.

USE OF PRINCIPAL COMPONENTS ANALYSIS TO DESCRIBE WATER QUALITY IN THIRTEEN STREAMS IN CENTRAL AND WEST-CENTRAL ALABAMA. Rosine W. Hall and John M. Aho, Auburn University Montgomery, Montgomery, AL 36124.

The Alabama Department of Environmental Management (ADEM) received a grant from the U.S. Environmental Protection Agency (EPA) to conduct a study of nutrient loading to Alabama reservoirs, and contracted with Auburn University Montgomery (AUM) to conduct a part of the study. This document presents the results of the AUM study of thirteen streams in central Alabama. Sampling occurred between December 8, 1998 and December 22, 1999. A total of 300 water samples were collected. Measured parameters include: water temperature, pH, specific conductivity, dissolved oxygen, turbidity, total Kjeldahl nitrogen, total ammonia nitrogen, total nitrate and nitrite nitrogen, total phosphorus, total suspended solids, and total dissolved solids.

Using a multivariate statistical technique called principal components analysis, we were able to rank the watersheds based on their relative overall water quality. In general, urbanized watersheds had lower water quality, but a few largely rural watersheds had much worse water quality relative to all sites we sampled than we would have expected.

ROTIFER DIVERSITY IN PONDS AND LAKES AT TROY UNIVERSITY. Eric S. Harris and Stephen C. Landers. Department of Biological and Environmental Sciences, Troy University, Troy, AL 36082.

Numerous ponds and lakes are located on the Troy University campus, which provide an excellent resource for teaching and research materials. For the past two years a survey of aquatic life has been conducted to catalog the protists and microscopic metazoans found in these freshwater bodies. This report will focus on the rotifers (Phylum Rotifera). Collections were made using a 20 [micro]m plankton net, which was cast from a dock or the shoreline and pulled below the surface. Organisms were immediately examined and were sketched, measured, and photographed when possible. Eleven different genera of rotifers have been identified. Of these, Keratella and Philodina were the most abundant, while the least abundant were Collotheca and Conochiloides. This project was supported by a Troy University Chancellor's Fellowship awarded to ESH.

SEASONAL VARIATIONS OF REPRODUCTIVE FUNCTIONS IN MALE MOSQUITOFISH, GAMBUSIA AFFINIS. Paul D. Melvin, III and Robert A. Angus, Dept. of Biology, Univ. of Ala. at Birmingham, Birmingham, AL, 35294.

The mosquitofish, Gambusia affinis, is a useful biomonitor for endocrine disrupting compounds (EDCs) because of its widespread presence around the world, large environmental tolerances, and ease of use in the laboratory. Previous studies have shown that vitellogenin production in males and modified anal fin development (masculinization) in females are reliable indicators of endocrine disruption. I hypothesize that sperm production in male G. affinis will also be a reliable indicator of EDCs and have shown in previous work that G. affinis living below a Waste Water Treatment Plant (WWTP) outfall have decreased mobilized sperm production. In order to further evaluate this hypothesis, it is important to understand any factors that cause natural fluctuations in sperm counts. This current study demonstrates the seasonal variability in mobilized sperm production in a population of G. affinis not exposed to endocrine disruptors. Male G. affinis were collected once per month for one year from a local, spring fed pond and brought back to the laboratory where sperm was collected and counted. Sperm production was fairly constant throughout the year (mean = 9.86 x [10.sup.6] [+ or -] 1.64 x [10.sup.6] SE) per ejaculate, with the exception of July, which had a significantly higher mean sperm count (2.15 x [10.sup.7] [+ or -] 1.87 X [10.sup.6], p < 0.001) than other months. Mean sperm counts for males from the nonpolluted spring sites were always significantly higher than from males collected at the site below the WWTP. These data contribute to the usefulness of sperm production by G. affinis as a biomarker of exposure to estrogenic EDCs.

DETERMINATION OF THE INCIDENCE OF CERVICAL DYSPLASIA IN AN HONDURAN USING TWO DIFFERENT TECHNIQUES. Thomas F. Garth, Dept. of Biology, BSC, Birmingham, AL 35254. George B. Inge, Dept. of Obstetrics and Gynecology, UAB, Birmingham, AL., Jeannette Runquist, Dept of Biol. BSC, Birmingham, AL 35254, and Luis Zuniga, San Pedro Sula, Honduras.

Under the direction of George B. Inge, the introduction of ThinPrep technology in San Pedro Sula, Honduras has provided the Our Little Roses home and indigent patients with a more effective cervical screening method. Numerous past studies indicate that Thinprep technology is both more sensitive and more accurate than conventional Pap smears in detecting dysplastic, pre-carcinomic, and carcinomic conditions, and therefore was accepted as the gold standard for this study. In order to measure the impact of Thinprep technology in a novel setting, results of both conventional Pap smears and Thinprep technology were recorded and analyzed along with the patient's relevant history. The data show that approximately 19.2 percent of patients (n=99) had incongruent results; therefore, indicating the presence of a false positive or a false negative result.

STRUCTURE AND FUNCTION OF FISH ASSEMBLAGES IN BEAVER PONDS OF THE CHOCTAWHATCHEE-PEA AND CONECUH RIVER WATERSHEDS, ALABAMA. Amanda F. McCall, Jimmy L. Aplin, Wes A. Robertson, and Paul M. Stewart, Department of Biological & Environmental Sciences, Troy University, Troy, AL 36082.

Beaver dams affect North American streams, altering ecosystem processes and influencing aquatic biota. Yet, little is known about fish assemblages in beaver ponds of the coastal plains of Alabama and their affects on stream fish assemblages. Twenty-five beaver ponds in the Choctawhatchee-Pea and Conecuh River watersheds of Alabama were examined. Ponds were sampled using both a Smith Root [R] Model 12-B backpack electrofisher and a seine. Water chemistry, area (ha), maximum depth (m), and mean depth (m) were obtained at each site. Biological indices were used to examine fish assemblage structure and function. One thousand two hundred sixty-five individuals and 29 species were collected. Lepomis macrochirus, Gambusia holbrooki, and Labidesthes sicculus were most common in beaver ponds and Ericymba buccata, Cyprinella venusta, and Lepomis megalotis were most common in streams. Number of species, percent tolerant individuals, number of intolerant individuals, number of darter and madtom species, and percent individuals as lithophilic species differed significantly from the same metrics observed in fish assemblages from stream environments in this region. This study suggests that beaver dams biologically alter stream fish assemblages, changing them both structurally and functionally.

PROTEIN ELECTROPHORETIC DISTRIBUTION OF GENETIC VARIATION IN SAUGER POPULATIONS. Amy Barr and Neil Billington, Dept. Biol. Environ. Sci., Troy Univ., Troy, AL 36082.

Sauger (Sander canadensis) is a carnivorous percid species of fish that are common in the mid-western U.S. and the Great Plains region. Little work has been conducted on genetic variation in sauger compared to the congeneric walleye (S. vitreus), which tends to be more popular with anglers. Genetic variation was screened by cellulose acetate electrophoresis for 991 saugers in 11 populations from the mid-west (South Dakota and Iowa) and the Great Plains (Saskatchewan and Alberta, Canada, Montana, and Wyoming). Two loci of 35 screened were polymorphic in sauger: super oxide dismutase (SOD-2*) and esterase (EST*). Genetic variation at SOD-2* was only detected in the Missouri River drainage populations. Highly significant among population heterogeneity was found at both EST* and SOD-2. Most populations in Montana and Wyoming showed significant deviations from Hardy-Weinberg expectations, all due to heterozygote deficits, likely caused by genetic drift or inbreeding related to small population sizes in these two states. Management agencies should try to maintain population sizes in Montana and Wyoming by reducing bag limits and encouraging catch and release of sauger.

VARIATIONS OF LIFE HISTORY AMONG BURROWING CRAYFISH IN THE CAMBARUS DIOGENES (GIRARD) COMPLEX. Jonathan M. Miller and Paul M. Stewart, Dept. of Biol. & Environ. Sci., Troy University, Troy, AL, 36081.

Limited information exists on crayfish (Decapoda: Cambaridae) in the southeastern United States. Information on primary burrowing crayfish is even more uncommon, as they are extremely difficult to collect. Morphological differences show at least two variations among the Cambarus diogenes (Girard) complex. The objective of this study was to determine and compare the life histories of two species (C. diogenes and Tubericambarus sp.) in the complex over a one-year period. Approximately ten crayfish were excavated monthly at three designated locations within the upper Choctawhatchee River (AL) watershed. Unfortunately, April, August, and September were not sampled. Specimens collected were identified and sex was noted along with the reproductive form of the male. We found a total of 142 crayfish of both species during the study. Sex ratios were in favor of the females for both species (C. diogenes 1.7:1; Tubericambarus sp. 1.3:1). There were fewer C. diogenes males found during the study (n = 12), 58% were Form 1 and 42% were Form 2. Due to the small sample size for this species, the appearance of Form 1 males were sporadic. More Tubericambarus sp. males were found (n = 48), of these 31% were Form 1. Form 1 males of this species were found from November through March, and two were found during July. Females of both species were found throughout the study (n = 82), but only one ovigerous Tubericambarus sp. was found during February.

UPDATE ON THE BACTERIOLOGICAL CONTENT OF DRY CREEK. Lydia Dews Glasscock and Dr. Brian S. Burnes, Biology Dept., Judson College, Marion, AL 36756

The distribution of resistance to ampicillin, chloramphenicol, erythromycin, neomycin, oxytetracycline, spectinomycin, vancomycin, tetracycline, and streptomycin among fecal coliforms was investigated as part of a continuation of an observation of the bacteriological content of Dry Creek in Perry County. Two years ago, this creek was shown to have a large amount of certain bacteria, including E. coli. The origins of these bacteria were then traced and found in some instances to be human in origin. Since this survey, a nearby trailer, thought possibly to be the point source of pollution responsible, has installed a septic tank. This followup has taken multiple samples of the creek, both during a dry spell and immediately after rain, attempted to culture fecal coliforms from the samples, and then checked for antibiotic resistant strains of the fecal coliforms cultured using these different antibiotics. All samples taken show considerable antibiotic resistance to all antibiotics save neomycin and streptomycin. Antibiotic resistance was also greater against ampicillin, oxytetracycline, and tetracycline after a period of rain than after a dry period. A human point source of pollution is still highly indicated by the levels of antibiotic resistant strains of fecal coliforms present in the waters of Dry Creek.

HYBRIDIZATION AND INTROGRESSION BETWEEN SAUGER AND WALLEYE DETERMINED BY PROTEIN ELECTROPHORESIS. Janet Gaston, Rachael N. Koigi, Ronald E. Creech, P. Taylor Ezell, and Neil. Billington, Dept. Biol. Environ. Sci., Troy Univ., Troy, AL 36082.

Sauger (Sander canadensis) and walleye (S. vitreus) are large predatory percid fishes that are highly prized by anglers. They are known to hybridize and these hybrids then backcross with the parental species leading to introgression, the movement of genes of one species into another. Several studies have shown that morphological examination is unreliable for identifying hybrid and introgressed fish compared to protein electrophoresis. Cellulose acetate gel electrophoresis was used to examine >2000 fish collected from lowa (one population), Montana (15 populations), South Dakota (three populations), Wyoming (two populations), and Saskatchewan, Canada (one population) to document hybridization and introgression between sauger and walleye. No hybridization was detected in Wyoming. In Pool 13 of the Mississippi River, Iowa, 25% of individuals were hybrid or introgressed. In Montana, between 0-22% of sauger from the Missouri River drainage and 0-10% from the Yellowstone River drainage were hybrids or backcrosses. In South Dakota, 4.2% of fish were hybrids or introgressed in Lake Sharpe, 3.9% in Lake Francis Case, and 21.2% in Lewis and Clark Lake. In Lake Diefenbaker, Saskatchewan, 20.9% of fish were hybrid or introgressed. In all cases hybrid numbers were underestimated by morphological examination. Protein electrophoresis was much more reliable than morphological examination for separating sauger, walleye, and their hybrids.

NORTHERN BLOT ANALYSIS OF DIFFERENTIALLY INDUCED TRANSCRIPTION TERMINATION REPORTER GENES. Yoon Cho, Dan Bai, and Christi Magrath, Department of Biological and Environmental Science, Troy University, Troy, AL 36082.

As demonstrated in previous enzymatic studies of transcription termination in Saccharomyces cerevisiae, levels of transcription termination from a galactose-inducible reporter construct correlate with the level of galactose present in the growth media. The reporter construct used in the termination assays contains a galactose promoter upstream of an intron-imbedded DNA sequence insertion site that is upstream from the gene for [beta]-galactosidase (LacZ). When termination occurs, the LacZ gene is not expressed and the RNA produced terminates with the inserted DNA sequence. Conversely, if termination does not occur, the LacZ gene is expressed and the inserted DNA sequence is removed from RNA by splicing. Previous enzyme assays established that the level of [beta]-galactosidase activity directly correlated to galactose levels; however, to demonstrate the direct molecular nature of this observation, yeast cells containing the reporter construct were grown in media with varying galactose concentrations. Total RNA was obtained, and the quality and concentrations were assessed using gel electrophoresis and spectrophotometry. Northern analysis can be used to directly assess the level of terminated versus non-terminated RNA within the differentially induced samples and to corroborate the use of using varying galactose concentrations as a method of directly altering termination levels.

GENETIC VARIATION IN MONTANA SAUGER AND HYBRIDIZATION WITH WALLEYE. Rachael N. Koigi, Jingyuan Xiong, Neil Billington, Dept. Biol. Environ. Sci., Troy Univ., Troy, AL 36082 and William. Gardner, Mont. Dept. Fish, Wildlife, &.Parks, Lewistown, MT 59457.

Sauger (Sander canadensis) populations have been declining in Montana. Factors blamed for this decline include over fishing, diversion dams, habitat degradation, and hybridization with introduced walleye (S. vitreus). Sauger were collected from 14 Montana populations, plus two populations from adjacent Wyoming, and one population from the Milk River, Alberta, Canada, that flows into Montana. Hybridization and introgression between sauger and walleye, and genetic variation in sauger were studied by cellulose acetate gel protein electrophoresis at four diagnostic loci. Hybridization rates were higher in the Missouri River (0-22%) than in the Yellowstone River populations (0-10%). Polymorphism was found at two (SOD* and EST*) of the 35 loci screened. Significant heterogeneity in allelic frequencies was documented in Yellowstone River and Missouri River populations at both loci. Several sauger populations showed significant deviations from Hardy-Weinberg expectations, all due to heterozygote deficits, likely caused by inbreeding or genetic drift. Montana sauger showed moderate population subdivision. Managers should try to minimize further reductions in population size by reducing bag limits and encouraging catch and release of sauger. Hybridization with walleye poses a serious threat to the genetic integrity of Montana sauger. Suggested management options include increasing bag limits for walleye where the two species co-occur.

UNDERSTANDING MOLECULAR CLONING AND THE STUDY OF GFAP EXPRESSION WITHIN ASTROCYTES. David V. Yokum, J. Runquist, Birmingham-Southern College, Birmingham, AL 35254, M. Brenner, J. Morello, and Y. Lee, Dept. of Neurobiology, Univ. of Ala., Birmingham, AL 35233.

Astrocytes are the most abundant cell type within the human central nervous system (CNS), and are potentially responsible for numerous developmental and regulatory functions within both normal and diseased states. In particular, astrocytes undergo reactive gliosis during CNS injuries in which they become enlarged and elevate levels of a variety of substances such as glial fibrillary acidic protein (GFAP). Although many of the consequences of reactive gliosis are likely to be beneficial, others have potential deleterious effects, and thus the ability to control GFAP expression could provide profound clinical utility. Therefore, this project seeks to understand the transcriptional regulation of the hgfa gene which encodes GFAP utilizing a reverse genetics approach. Essentially, the function of a specific hgfa DNA region is destroyed to determine whether its transcription is critical for controlling the astrocyte specificity and upregulation of GFAP in response to CNS injury. The development and isolation of such a mutated construct requires a series of molecular cloning techniques, and will be the focus of this presentation.

CARLSON'S TROPHIC STATE INDEX AS APPLIED TO SOUTHEASTERN ALABAMA FISH PONDS. P. Taylor Ezell and N. Billington. Dept. Biol. Environ. Sci., Troy Univ., Troy, AL 36082.

Carlson developed a Trophic State Index (TSI) in 1977 based on Secchi disk depth as a practical tool for measuring the algal productivity in water bodies, where values range from 0-100, each 10 unit increase represents a doubling in algal biomass. The index also has equations for converting chlorophyll a, total nitrogen and total phosphorus to TSI. Carlon's index was developed in Wisconsin, but has been applied to water bodies in other parts of the U.S. However, little work with TSI has been conducted in the southeastern U.S., where the majority of the water bodies are warm, monomictic fish ponds. Many of these man-made ponds are fertilized with nutrients, especially phosphorus to enhance fish productivity. We surveyed 30 water bodies (28 fish ponds and two reservoirs) in southeastern Alabama. At each water body, Secchi disk depth and a sub-surface water sample were collected during spring and fall overturn. Chlorophyll a and nutrient (nitrogen and phosphorus) concentrations were determined. Vertical profiles of conductivity, dissolved oxygen, light intensity and temperature were also recorded. Total alkalinity, hardness, pH, turbidity, and total dissolved solids were recorded also. The TSI values obtained from Secchi disk depth data were compared with those calculated from chlorophyll a and nutrient data. Values of TSI ranged from 48-93 and chlorophyll a concentrations ranged from 1.54-181.10 [micro]g/L. These southeastern Alabama water bodies ranged in productivity from oligotrophic to hypereutrophic.

ETHOXYRESORUFIN-O-DEETHYLASE (EROD) ACTIVITY IN CHANNEL CATFISH (ICTALURUS PUNCTATUS) EXPOSED TO TROY (ALABAMA) WASTEWATER TREATMENT PLANT EFFLUENT. Alicia Whatley, Smriti Subedi Bhattarai, Paul M. Stewart, Christi Magrath, and Philip Reynolds, Dept. Biol. & Environ. Sci., Troy University, Troy, AL 36082.

The effect on induction of cytochrome P450 enzymes, which are involved in the first stage of xenobiotic transformation, was examined following exposure of channel catfish (Ictalurus punctatus) to Troy (Alabama) Wastewater Treatment Facility effluent on Walnut Creek. After one day of exposure, results showed that peak induction levels, measured as ethoxyresorufin-O-deethylase (EROD) activity, were 3-fold higher in channel catfish exposed to wastewater from the mixing zone on Walnut Creek, compared to fish exposed to the water from upstream on Walnut Creek. Water from Big Creek (Barbour County, Alabama) was used as a reference unpolluted site (based on a higher Invertebrate Community Index obtained from a previous assessment) for administration of PCBs, which are a known inducer of cytochrome P450 enzymes. While there was no significant increase in EROD activity in Big Creek PCB treated over Big Creek control, EROD activity in Big Creek control was 2.7-fold higher than Walnut Creek control. Although previous assessments have focused on either water quality or lethal effects on organisms, sublethal effects, such as EROD activity, might be a more sensitive indicator of the effects of wastewater and other contaminants on aquatic organisms. More water bodies in the area, even those thought to be of good quality with no apparent contaminant source, should be examined for contaminant-related cytochrome P-450 activity.

GOPHER TORTOISE (GOPHERUS POLYPHEMUS) POPULATIONS ON STATE LANDS IN MOBILE AND BALDWIN COUNTIES, ALABAMA. Lindsey A. Timmerman, David H. Nelson, and Joel A. Borden. Dept. of Biology, University of South Alabama, Mobile AL 36688.

Once having a geographic distribution that extended from South Carolina to Louisiana, the gopher tortoise (Gopherus polyphemus) now is found mostly in small, fragmented, isolated populations. Gopher tortoises have been studied extensively in some portions of their historic range, such as Florida, where they are relatively abundant. Other areas of the Southeast, however, have been largely neglected. Very few studies have been conducted in southern Alabama for the past fifty years. In an effort to fill this information gap, two gopher tortoise populations were studied in Alabama (Jacinto Port, Mobile Co. and the Northeastern sector of the Mobile-Tensaw Wildlife Delta Management Area, Baldwin Co.). We sought to assess how elevation, soil type, and vegetation might influence major population parameters: population density, age structure, reproductive success, and mobility. The trapping program yielded a total of 92 separate captures for the entire trapping season. Thirty-five individual tortoises were captured at the Northeastern Management Area: 29 males, 4 females, and 2 juveniles. Twenty-two individual tortoises were captured at Jacinto Port: 12 males, 9 females, and 1 juvenile. In Jacinto Port, the male to female ratio was 1.3 males for each female, and in the Northeastern Management Area the male to female ratio was 7.2 males for each female. During the study, we also tested both populations for Upper Respiratory Tract Disease. Twenty-eight distinct blood samples were collected over the course of the trapping season, and all results were negative.

NEW AND INTERESTING PLANTS FROM ALABAMA. Michael Woods and Alvin R. Diamond, Department of Biological and Environmental Sciences, Troy University, Troy, AL 36082.

Eleven species are reported as new to the flora of Alabama. Additionally, six significant new county records are reported for five other taxa. Of the 16 total taxa reported, four are seedless vascular plants, eight are eudicots and four are monocots. Of the eight taxa non-native to the United States, three, Begonia cucullata Willd., Marsilea minuta L., and Pistia stratiotes L. have invasive potential.

Distribution and Abundance of the Mississippi Diamondback Terrapin (Malaclemys terrapin pileata) Along Coastal Mobile County, Alabama. David H. Nelson and Joel A. Borden, Biological Sciences, University of South Alabama. Mobile, AL 36688.

Diamondback terrapins were sampled from April to August 2004 and from May to August 2005, along the southwestern coast of Alabama (Mobile County) from Bayou Heron to Cedar Point. Several trapping techniques were utilized: pitfall traps, modified crab traps, hoop traps, box traps, and nest depredation surveys. Terrapins were individually marked (notching marginal scutes), weighed, and measured (carapace length, carapace width, shell height, plastron length, tail length, and head width). Females were palpated for eggs; anomalies were noted (predation marks, prop scars, barnacle and oyster fouling), and all individuals were digitally photographed. Our goal was to systematically document the abundance and distribution of terrapins in Alabama coastal waters. Twenty-eight adults were captured (19 females, 8 males, 1 juvenile) over the two sampling seasons. Eight hatchlings were also encountered, although two were found dead. Terrapin populations appear to be sparse and isolated along coastal Alabama. The terrapin is currently listed as a "Priority One" (highest conservation concern) by the Alabama Department of Conservation and Natural Resources. Further research is needed to identify current threats to terrapin populations in Alabama and to provide for the conservation of the species in its natural habitat.

ANTIMICROBIAL ACTIVITY OF ANTARCTIC MARINE SPONGE EXTRACTS. Kevin J. Peters, Charles D. Amsler, James B. McClintock, Dept. of Biology, UAB, Birmingham, AL 35294-1170. Bill J. Baker, Dept. of Chemistry, USF, Tampa, FL 33620.

Sponges often have a great number of bacteria and other microorganisms associated within the (up to 40% of the biomass). Previous studies have documented symbiotic relationships between sponges and their associated microorganisms with some providing nutrition, protection against UV radiation, and/or secondary metabolite production. Conversely, some microorganisms may be pathogenic to sponges, or may be species-specific in their beneficial roles. Many sponges are known to be chemically defended against large predators (e.g. echinoderms and fish) and sponges may also be defended against sympatric, non-beneficial microorganisms. This study will identify the in vitro bioactivity of crude polar and non-polar extracts of antarctic marine sponges against a suite of sympatric bacteria. The bacteria were isolated from the tissues of several different antarctic marine organisms and associated water. In this ongoing study, bioactivity is measured by the ability of the extracts to prevent microbial growth on marine agar plates.

A Comparison of Herpetofaunal Communities Between Two Upland Mosaic Habitats Bordering the Mobile-Tensaw Delta. Joel A. Borden and David H. Nelson, Biological Sciences, University of South Alabama, Mobile, AL 36688.

A herpetofaunal field inventory was conducted to assess how habitat variation influences species diversity, abundance, and distribution in upland coastal plain ecosystems. The study sites were two diverse, upland areas bordering the Mobile-Tensaw Delta in southwestern Alabama. Sampling continued from February to December 2005. The objectives were to examine community structures of amphibians and reptiles (with respect to habitat type) on state-owned lands in Baldwin and Mobile Counties, Alabama. Ten representative habitat types were selected, based on elevation, dominant vegetation, age of stand, ground cover, percent canopy cover, soil type, and hydrology. The Baldwin County site is dominated by a 20-year old, former loblolly pine plantation with elevations ranging from 7-30 meters. The Mobile County site is characterized by oak flatwoods interspersed with long-leaf pines between 60 and 80 years old, at an elevation of 3 meters. Each site was sampled by a single drift fence array (composed of three arms) with six terrestrial funnels and three 19-liter pitfalls. Also included in each array were twelve cryptozoan covers, five PVC tubes (treefrog refugia), visual transects, anuran call monitoring, and hand captures. Twenty-one species of amphibians (N=2447) and thirty-one species of reptiles (N=916) were encountered. Our results confirm that different habitat types profoundly affect specific herpetofaunal assemblages. Proper management and the designation of adjacent refuge habitats surrounding a monoculture stand may mitigate the loss of herpetofaunal diversity.

INNER WORKINGS OF EQUUS CABALLUS: THE EFFECTS OF COMMON-USAGE MEDICATIONS ON THE NATURAL FLORA OF THE EQUINE GASTROINTESTINAL TRACT. Dr. Brian Burnes, Dept. of Biological Sciences, Judson College, Marion, AL 36756, Christy R. Spearman and Dr. Brian Burnes, Judson College, Marion, AL 36756

The natural bacteria within the equine intestinal tract are often overlooked during use of certain medications in consideration of the positive effects. The purpose of this experiment is to demonstrate the effects of three common-usage equine medications on Escherichia coli, Lactobacillus acidophilus, and Bacillus cereus. These three bacteria are just a few of the most common bacteria found in the gastrointestinal tract, therefore being the most logical with which to work. Separated in three agar dishes, each bacteria is exposed to a realistic dilution of each medication. Results demonstrate the effect each medication has on the different bacteria, providing a rational idea of the possible damage within the equine intestinal tract.

DNA SEQUENCE ANALYSIS OF A PUTATIVE CYTOCHROME P450 FROM WASTEWATER TREATED CHANNEL CATFISH. Robert McHugh, Smriti Bhattarai, Christi Magrath and Alicia Whatley, Department of Biological and Environmental Science, Troy University, Troy, AL 36082.

In the channel catfish, activation and expression of cytochrome p450 is evident within three days of exposure to wastewater treatment effluent. Maximal levels of p450 expression occur after six days then slowly return to basal levels. In previous analysis, both RT-PCR and EROD activity were used as markers of activity within the liver. RT-PCR of liver RNA using primers complementary to known p450 sequences from fish species generated DNA products of the expected size as determined by gel electrophoresis; however, DNA banding patterns indicated the presence of two unexpected DNA products. Consequently, the putative p450 DNA band, as well as the two unidentified DNA products, were purified from 1.5% agarose for use in DNA sequencing. The RT-PCR product's identities can be confirmed by comparative DNA analysis.

TRANSCRIPTION TERMINATION CAPACITY OF INTERGENIC REGIONS FROM S. CEREVISIAE. Pranjal Nahar, Kevin Flinn, James Noble, and Christi Magrath, Department of Biological and Environmental Science, Troy University, Troy, AL 36082.

Chromosome III of Saccharomyces cerevisiae contains twenty Autonomous Replication Sequences (ARSs). The ARS elements are localized to primarily intergenic regions and the most replicatively active ARSs are found less than 200 base pairs from the 3'-end of a gene. It is hypothesized that the ARS localization is due to the protective nature of the transcription termination sequences found at the 3'-end of the genes; thus, the transcription termination sequences act as "molecular shields" to protect the ARSs. To evaluate this hypothesis a screen of the intergenic regions harboring the ARS elements is in progress. As part of this screen, a set of oligionucleotide primers with specificity to the termini of the intergenic regions was designed. These primers can be used to amplify the intergenic regions and to generate reporter constructs that can be used to assess transcription termination capacity with enzymatic assays, ultimately allowing correlation between replication and transcription termination capacity. This study advances the understanding of the function of transcription termination elements and will be one of the first transcription termination profiles.

COMMELINA ERECTA--ITS FLOWER STRUCTURE, REPRODUCTIVE BIOLOGY, AND ITS INTERACTIONS WITH HERBIVORES. Roland Dute and Debbie Folkerts, Dept. of Biological Sciences, Auburn University, AL 36849.

Commelina erecta (erect dayflower) is a species native to much of the eastern and southern U.S. It flowers from mid-June through early September in East Central Alabama. The inflorescence develops within a spathe or modified leaf. Flowers emerge one at a time from the spathe every other day to every fourth day. An individual flower is open for only a matter of hours. The petals then deliquesce and the pedicel curves such that the developing fruit is pulled back into the spathe where fruit development continues. The spathe contains a mucilage whose complete chemical composition is unknown but includes the elements silicon, calcium, chlorine, and potassium. A common predator of C. erecta is the beetle Neolema sexpunctata whose larvae feed on the underside of the leaves. The larvae tend to avoid the outer surface of the spathe probably due to the high concentration of siliceous hook-shape trichomes found there. Grasshoppers also are a major predator of leaves and spathes. Counts of flower stalks pre- and post-anthesis show a considerable loss of reproductive potential via flower abscission. The proximal abscission scars show features identical to similar scars found in dicotyledonous plants. Insect visitors such as syrphid flies (Toxomerus boscii) and bumblebees (Bombus spp.) are common visitors and carry pollen grains of C. erecta on their bodies. Although C. erecta can self-pollinate, insect visitors provide cross pollination.

PIT MEMBRANE STRUCTURE IN THE WOOD OF TWO SPECIES OF GYMNOSPERMS. LaToya Hagler, Adam Black, and Roland Dute, Dept. of Biological Sciences, Auburn University, AL 36849.

Pit membranes provide a pathway for the flow of water between tracheids in the xylem of vascular plants. The torus, a central thickening of the pit membrane (as distinct from the thin margo), interacts with the pit apertures to provide a safety mechanism that impedes the movement of air embolisms throughout the vascular system. This study is designed to compare the structure and development of pit membranes in two species of cone-bearing gymnosperms, Metasequoia glyptostroboides and Abies firma. Branch segments were preserved and processed for microscopy by traditional methods. Thin sections of xylem were stained and viewed both by light and electron microscopy. In both species, the torus thickening is initiated at an early stage in the ontogeny of the pit membrane. Pit membranes of Abies are traversed by secondary plasmodesmata. Thus far, no such structures have been located in Metasequoia. As the tracheid cytoplasm undergoes autolysis, the matrix materials in pit membranes of Metasequoia are removed from both torus and margo regions. However, in Abies only the margo loses its matrix material, whereas the torus remains intact. It is apparent from the photographs that the torus in mature, water-conducting tracheids of Abies is a more substantial structure than the torus of Metasequoia. Whether this difference in final structure leads to a difference in protection from the spread of air embolisms is unknown. Work is continuing on these species in an effort to correlate the differences in pit membrane ontogeny with cytoplasmic differences during development.

HABITAT USE BY PLETHODONTID SALAMANDERS IN NORTHEAST ALABAMA. Clifford J. Webb and Dr. George Cline, Dept. of Biology, Jacksonville State Univ., Jacksonville, AL 36265.

Plethodontid salamanders form complex communities in small streams. This fifteen-month study examines physiological/morphological trends, habitat usage, seasonal activities and reproductive phenology of seven Plethodontid salamanders. The following salamanders were collected: Eurycea cirrigera, Eurycea guttolineata, Desmognathus conanti, Plethodon glutinosus, Plethodon serratus, Gyrinophilus porphyriticus and Pseudotriton ruber. The study site is in the North-East corner of Alabama, in Talladega county (33[degrees] 32'57.4"N, 85[degrees]49'23.7"W) in Talladega National Forest. Three reaches in a seasonally intermittent stream were studied. Each reach contained three different hydro geomorphic units with one repeat per reach. Three open-ended PVC pipe traps and leaf-litter bags were placed in each of the geomorphic units at each site. Eurycea cirrigera was by far the most abundant salamander collected with all developmental stages observed. Adult Eurycea guttolineata were encountered infrequently, but they were always found on stream margins. Desmognathus conanti were second in abundance to E.cirrigera. Plethodon glutinosus were always found in the riparian zone under debris or in leaf litter bags. Plethodon serratus only appeared in cold, wet winter months and always under debris away from the stream. Gyrinophilus porphyriticus were always found in the stream and only as larvae. Pseudotriton ruber appeared in very low numbers and only terrestrial adults were found usually under rotted logs. E.cirrigera and D.conanti were in highest abundance in riffle areas. Larval E.cirrigera, D.conanti and G.porphyriticus were identified from the stream. All larvae collected were also in highest abundance in riffle areas.

BENTHIC FEEDING BEHAVIOR IN THE COASTAL LOBATE CTENOPHORE MNEMIOPSIS MCCRADYI. J. Christopher Taylor, Deborah C. Odom, and Anthony G. Moss, Biological Sciences, Auburn University, 36849.

Mnemiopsis mccradyi is a common coastal plankter in the Gulf of Mexico. We describe here observations that indicate that Mnemiopsis does not limit its feeding to the water column. We routinely collect ctenophores with sediment in their stomodea and with benthic materials (such as benthic diatoms) in their food grooves. In lab, we observe M. mccradyi to interact with the flocculent layer of sediment using 8 distinct behavioral modes. Video recordings show Mnemiopsis moving sediment via precise lobular contractions combined with auricular groove currents generated by auricular comb plates. Particles captured from the spiral auricular current are passed directly to the food groove. Soluble fluorescein was used to track current flow through the interlobular volume and the auricular grooves in animals resting with the tips of their lobes on the flocculant upper sediment. As Mnemiopsis pushes against the sediment, auricular currents draw water to and around the animal and through the interlobular spaces. This study reveals a heretofore-unrecognized mechanism for carbon transfer into the water column, and could provide a mechanism for the acquisition of a recently-discovered protistan parasite assemblage (Hydrobiologia 451: 295). NSF Grant # NSF-MCB-0348327 to AGM, REU to JCT.

GAMETOPHYTIC AND SPOROPHYTIC RESPONSES OF PTERIS SPP. TO ARSENIC. T. Justun Durham and Hollings T. Andrews, Dept. of Biology, Tenn. Tech. Univ., Cookeville, TN 38505. T. Wayne Barger, Dept. of Conservation, State Lands Division, Montgomery, AL 36130. Matthew S. Wilson, Florida Dept. of Environmental Protection, Bureau of Mine Reclamation, Tallahassee, FL 32310.

Few plant species have demonstrated the ability to hyperaccumulate heavy metals from contaminated soil. Recently, Pteris vittata L. has been identified as a hyperaccumulator of arsenic. Because gametophytic development is an essential stage in the fern life cycle, impacts of heavy metal hyperaccumulation on gametophytic and sporophytic tissue must be investigated if successional bioremediation efforts are to be implemented successfully. Our research showed that sporophytes as well as gametophytes of P. vittata are capable of As uptake and accumulation. Increased As ([less than or equal to]2500 ppm) did not inhibit spore germination, and deleterious effects on gametophyte morphology were observed only after extended time periods on media with extremely high As concentrations ([greater than or equal to]600 ppm). Six other Pteris species varied in ability to germinate on As-containing media. Sporophytes of P. vittata showed no adverse effects when exposed to the highest soil As levels (1650 ppm); in fact, root proliferation was observed in areas of increased As concentration (250 ppm). Foliar application of an arsenical herbicide (calcium acid methanearsonate) to sporophytes resulted in decreased chlorophyll and carotenoid concentrations. Phosphate additions inhibited As uptake by sporophytes, indicating As uptake involves the phosphate transport system.

ECONOMIC UTILIZATION OF POTATO PEEL WASTE FOR BIOREMEDIATION OF PERCHLORATE IN WATER. Benedict C. Okeke*, Dept. of Biology, Auburn University Montgomery, AL 36124-4023. William T. Frankenberger Jr., Dept. of Environ. Sci., Univ. of California Riverside, CA 92521.

Perchlorate (Cl[O.sub.4.sup.-]) is detected in ground water throughout the United States, because Cl[O.sub.4.sup.-] was extensively used in solid propellants for rockets, missiles and explosives. Cl[O.sub.4.sup.-] is recalcitrant in the environment and is potentially toxic to various forms of life. Bioremediation is an attractive strategy for Cl[O.sub.4.sup.-]-contaminated water. Carbon substrates such as acetate used for microbial reduction of Cl[O.sub.4.sup.-] are, however, expensive. This study thus explored the potential application of potato peel waste and starch in combination with an amylolytic bacterial consortium for Cl[O.sub.4.sup.-] bioreduction in water. Potato peel waste supported Cl[O.sub.4.sup.-] reduction by Dechlorosoma sp. perc1ace with the rate of Cl[O.sub.4.sup.-] reduction being dependent on the amount of potato peels. Cl[O.sub.4.sup.-] was rapidly degraded reaching non detectable level (<4 [micro]g/L) in 9 days. No substantial reduction of Cl[O.sub.4.sup.-] was observed in sterile potato peel medium without Dechlorosoma sp. perc1ace in 7 days. Redox potential of the potato peel cultures was favorable for Cl[O.sub.4.sup.-] reduction, decreasing to as low as -294 mV in 24 h. Sugar levels remained very low in cultures effectively reducing Cl[O.sub.4.sup.-] and was substantially higher in sterilized controls. Results indicate that potato peel waste in combination with amylolytic and Cl[O.sub.4.sup.-] reducing bacteria can be economically used to achieve complete Cl[O.sub.4.sup.-] removal from water.

EFFECTS OF ENVIRONMENTAL STRESS ON LEVELS OF CYANIDE PRODUCTION IN SELECTED PLANTS. Anna Chandler and H. Wayne Shew, Dept. of Biology, Birmingham-Southern College, Birmingham, AL 35254.

More then 2,650 species of plants have been found to be cyanogenic or cyanide producing. Cyanogenesis in plants results from the action of plant enzymes that convert complex cyanogenic glycosides into hydrogen cyanide (Asano, 2005). Cyanogenesis functions to reduce damage to plants from attack by fungi and herbivores, and occurs when cells are damaged or stressed. In this study, we exposed the leaves of a variety of cyanogenic plants to environmental stress conditions. These conditions included salinity, drought, herbicide application, high nitrogen fertilizer application, and mimicked herbivory. We then used a quantitative, spectrophotometric method to measure the amount of cyanide produced in the leaf tissues of these plants. We found that in some species certain stress conditions led to an increase in the level of cyanogenesis.

THE EFFECT OF DIETARY CARBOHYDRATE ON WEIGHT GAIN AND GONAD PRODUCTION IN JUVENILE LYTECHINUS VARIEGATUS (ECHINODERMATA). Anna L. Morris, Mickie L. Powell, and Stephen A. Watts, Dept of Biology, Univ. of Ala. at Birmingham, Birmingham, AL 35294. Addison L. Lawrence, Texas Agricultural Experiment Station, Texas A&M, Port Aransas, TX 78373.

The high demand for sea urchin gonads (roe or uni) has resulted in overfishing of many wild sea urchin populations. To supply the demand for roe, the development of a sea urchin aquaculture industry using cost-effective formulated feeds is necessary. In adult sea urchins, formulated feeds support weight gain and gonad production; however, studies demonstrating the effects of formulated feeds on juvenile growth are limited. In the first study, juvenile sea urchins as small as 3 to 4 mm test diameter consumed and utilized a semi-purified feed formulated for sea urchins. In a second eight-week study, feeds containing 31% protein (dry weight) and three levels of dietary carbohydrate: 1) low (LC, 16% dry weight); 2) medium (MC, 22% dry weight); and 3) high (HC, 36% dry weight), were fed to juvenile sea urchins reared individually at 22 C and 32 ppt synthetic seawater. Sea urchins fed the LC feed had significantly higher weight gain than individuals fed the HC feed. Weight gain of sea urchins fed the MC feed was not significantly different from LC or HC. Gonad production was minimal and did not vary with feed. These data indicate that adequate energy was available to support weight gain in all feeds; however, those individuals fed the highest protein:energy ratio had the highest weight gain. We suggest cost-effective, high quality feeds will support juvenile sea urchin growth and will be important in the developing aquaculture industry.

Keyword: Lytechinus variegatus

STATUS OF CANDIDATE MUSSEL SPECIES IN THE GULF COASTAL PLAINS, SE ALABAMA. Paul M. Stewart, Dept. Biol. & Environ. Sci., Troy Univ., Troy, AL 36081. Megan M. Pilarczyk, Dept. Biol., Wake Forest Univ., Winston-Salem, NC 27106.

Freshwater mussel assemblages in the southeastern United States are diverse yet imperiled. Continued status updates for this group are required for their preservation. In summer 2004, we performed a qualitative survey of freshwater mussel assemblages at 24 sites in southeastern Alabama and one site in northwest Florida. Efforts were focused on seven candidate species: Margaritifera marrianae, Fusconaia escambia, Lampsilis australis, Pleurobema strodeanum, Ptychobranchus jonesi, Quincuncina burkei, and Villosa choctawensis. The survey information was compared to recent historical records from the 1990s using presence-absence analysis. There was no significant difference between the number of taxa found in the 1990s and in 2004. There was a significant reduction from the 1990s to 2004 in the number of candidate species found at a site, with all candidate species except P. jonesi found at fewer sites. Pleurobema strodeanum declined at the greatest number of sites, making it a species of utmost concern. The historical data often lacked abundance and number of man hours information, thus it was difficult to make direct comparisons between the two data collection periods. Future mussel surveys should follow protocols such as those prepared by Carlson (et al.) and report the number of species, number of individuals of each species (live and dead), and number of man hours. Data collected in this study supports the need to elevate these species under Endangered Species Act protection.

ANALYSIS OF CILIARY AND FLAGELLAR ACTIVITY WITH A LASER SCATTERING MICROSCOPE. Anthony Moss and Margaret West, Biological Sciences, Auburn University, 36849

We have modified a metallurgical microscope to provide incident laser illumination of opaque ciliated epithelia to allow computer-aided analysis of ciliary beating. A Melles Griot 5 mW HeNe laser model MELU12 mounted to the side port of a Leitz Metallux ND microscope provided coherent, monochromatic laser light to the specimen via the outer illuminating tube of the Ultropak vertical illuminator and attached objectives (22X to 62X objectives worked well). Backscattered light was collected by a sensitive photodiode (model ST386-18L, Hamamatsu Corporation) mounted at the focal point of a 10X projection ocular. The output of the photodiode was sent to a differential amplifier operating at 1000-5000 gain (model 502, Tektronix) and the amplified output sent to an analog to digital converter (model 1401, Cambridge Electronic Designs). Digitized time-series data analyzed by Fast Fourier Transform revealed principal components that corresponded precisely to known frequencies applied to a reflective piezoelectric probe driven by a Hewlett-Packard sine wave generator at up to 500 Hz. The laser backscatter system reliably revealed the beating activity of an opaque ciliated epithelium removed from the Asian Clam, Corbicula fluminea at rates reported in the literature, namely 12 to 14 cycles per second. Surprisingly, the system was not directionally sensitive and could reveal beating at any angle of applied laser illumination. Development of this instrument allows improved, reliable and objective analysis of mucociliary transport systems with wider bandwidth than competing video-based systems. Support: USDA CSREES ALA016-033, NSF-MCB-0348327 to AGM.

SPONGE-PHOTOSYMBIONT ASSOCIATIONS IN CARIBBEAN CORAL REEFS OF PANAMA. Patrick M. Erwin and Robert W. Thacker, Dept. of Biology, Univ. of Ala., Birmingham, AL 35294-1170.

Symbioses between marine sponges and photosynthetic organisms have been described from coral reef communities; however, the frequency of these associations and the ecological nature of interactions between hosts and symbionts often remain unresolved. We determined the number and abundance of sponge species harboring photosymbionts in the shallow-water reefs of Bocas del Toro, Panama, by conducting line-intercept transects with measurements of chlorophyll a (chl a) concentration. Sponges were dominant members of these benthic communities, second in abundance only to stony corals. Twenty-five of the 67 species investigated (37.3%) exhibited high chlorophyll levels (>150 [micro]g/g) and accounted for 35.6% of the observed sponge community. Association with photosymbionts is a common strategy among coral reef sponges and may represent an important source of primary productivity in oligotrophic coral reef environments. Additionally, two sponge species known to harbor cyanobacterial symbionts (Aplysina fulva, Xestospongia subtriangularis) were experimentally shaded to test the effects of reduced light availability on symbiont load and host growth. Six weeks of shading reduced (>40%) the abundance/activity of symbionts. In A. fulva, control sponges exhibited over twice the growth of shaded sponges. In X. subtriangularis, no significant difference in growth was observed between control and shaded sponges. These results suggest that the relationship between sponges and their cyanobacterial symbionts varies among host species, with some sponges strongly dependent on symbiont photosynthesis and others unaffected by short-term decreases in symbiont activity.

PHOTODYNAMIC DYE TREATMENT OF FISH PATHOGENS. Mark Meade, Benjie Blair and Charles Olander, Jacksonville State University, Dept. of Biology, Jacksonville AL 36265.

Current treatments for pathogenic fish protozoa include using medicinal baths containing formalin, copper sulfate, malachite green, and/or combinations of the three. The stringency of these treatments can themselves result in fish demise. Initial medicinal bath experiments suggested that the photoactive dye phloxine b could be used to treat protozoa infestations including Tetrahymena and Ichthyophthirius without harm to the treated fish. Concentrations of 10 mg/L resulted in protozoa demise and reduced the numbers infesting fish. Recently the photoactive dye was compared with several other dye compounds, including acridine orange, rose bengal, methylene blue, eosin yellow, and malachite green. Tetrahymena were grown in light and dark conditions in culture media containing either 0, 1, 5, or 10 ppm of the dyes and held in an environmental chamber at temperatures of 19, 22, 26, or 30[degrees]C. Culture densities were monitored by turbidity measurements after 24 hr. In general phloxine b was more effective than the other dyes at reducing cell proliferation in the light. Malachite green, a non-photoactive dye, was the most effective treatment in the dark, however, phloxine b was as effective as malachite green in the light. Increased temperature resulted in reduced cell growth for all compounds tested. Overall, these data further demonstrate that the photodynamic activities of phloxine b may be useful in eradicating pathogenic fish protozoans.

FURTHER EVALUATION OF A PUBLISHED PCR TECHNIQUE IN TRACKING THE SOURCE OF FECAL CONTAMINATION IN ALABAMA AND WISCONSIN ENVIRONMENTAL WATERS. D. Salte[r.sup.1], M. Arringto[n.sup.1], Q. Sonnie[r.sup.1], G. Goldensta[r.sup.1] and M. Leuthe[r.sup.2], [.sup.1]Depart. Biol. Environ. Sci., UWA, Livingston, AL 35470; [.sup.2]Leuther Laboratories, Coon Valley, WI 54623.

We previously reported our results on using a published PCR technique based on species specific Bacteriodes species (Appl. Environ. Microb., 2000, 66: 4571) for tracking fecal contamination in Alabama environmental waters (Poster Presentation, AAS 2003 Annual Meeting). In summary, analysis of environmental waters around Livingston, AL for human versus ruminant fecal contamination showed that all 8 sites were positive for Bacteriodes DNA; Five and 4 sites were positive for human and ruminant Bacterioides DNA, respectively. Research was suspended until we could eliminate reagent/equipment DNA contamination problems. We hopefully have solved these problems by physically separating the DNA extraction and preparation of the reaction mixes from the PCR instrument and gel electrophoresis analysis steps as well as using dedicated micropipettors at each location. We now report confirmation of our previous results as well as more extensive analysis of control samples. In addition, we also surveyed one of the human fecal-contaminated streams at 10 sites from the beginning of the stream until its end into the local river. All but 1 of the sites were positive for Bacteriodes DNA with 8 and 7 sites positive for human and ruminant Bacteriodes, respectively. This technique was also used to analyze environmental waters collected weekly from several sites along a Wisconsin river during Summer 2005. Out of 143 samples, 110 were positive for ruminant Bacteriodes DNA and only 11 were positive for human Bacteriodes DNA. Nine samples were positive for both. Further research will be reported.

THE COMBINED APPLICATION OF SONICATION AND ADVANCED CHEMICAL OXIDANTS AS A METHOD TO TREAT INTRODUCED AQUATIC MACROINVERTEBRATES TRANSPORTED VIA BALLAST WATER. Meghana Gavand*, James B. McClintock*, Charles D. Amsler*, Robert W. Peter[s.sup.+],* Dept. of Biology, Univ. of Alabama at Birmingham, AL 35294. [.sup.+]Dept. of Civil and Environmental Engineering, Univ. of Alabama at Birmingham, AL 35294.

Introduction of invasive marine and freshwater invertebrates constitutes a growing threat to aquatic ecosystems. Ballast water is a major factor contributing to the introductions of aquatic invasive species. Three different life-history stages (cysts, nauplius larvae, and adults) of brine shrimp Artemia salina were chosen as model invertebrate life stages to determine the effect of individual and combined treatments of sonication and advanced chemical oxidants under static and continuous flow regimes. The sonication frequency, advanced chemical oxidation concentrations and treatment times were optimized based on previous studies with bacteria. The highest percent mortality obtained for individual treatments over a 5 min exposure was 17, 49 and 43% (static regime) and 16, 34 and 38% (continuous flow regime) for cysts, larvae and adults, respectively. The highest mortality obtained for combined treatments over a 5 min exposure using combined sonication, hydrogen peroxide and ozone was 83, 100, 100% (static regime) and 67, 100 and 100% (continuous flow regime) for cysts, larvae and adults, respectively. The combined treatments are synergistic and result in significantly greater mortality than individual treatments. Combined sonication and advanced oxidants may be an effective method to inactivate a variety of different life history stages of marine and freshwater invertebrates in ballast water. Supported by a grant from the US Fish and Wildlife Service to R. W. Peters, C. D. Amsler and J. B. McClintock.

SUMMER RESTING AREAS OF THE GULF STURGEON, ACIPENSER OXYRINCHUS DESOTOI, IN THE CONECUH/ESCAMBIA RIVER SYSTEM. Jon B. Sawyer and Paul M. Stewart, Department of Biological and Environmental Sciences, Troy University, Troy, AL 36082. Frank M. Parauka, U.S. Fish and Wildlife Service, Panama City Field Office, Panama City, FL 32405.

Once inhabiting Gulf coastal rivers from the Mississippi River to Tampa Bay, the Gulf sturgeon Acipenser oxyrinchus desotoi is now rarely found south of the Suwannee River in Florida. Among the Florida Panhandle rivers, the Conecuh/Escambia River contains the most suitable Gulf sturgeon habitat. Radio telemetry was used to track thirteen Gulf sturgeon within the Conecuh/Escambia River, during April through November of 2005, to locate sturgeon summer resting areas and to observe movement behavior. Only one radio-tagged sturgeon moved upstream into the Conecuh River, with the remaining study population observed in the Escambia River, below the Florida line. Three summer resting areas within the Escambia River were identified, and one previously located resting area was confirmed during this study. Movements within the summer habitats did not appear to be influenced by abnormally elevated flow volumes observed during this study. Substrates for the summer resting areas were recorded as primarily coarse sand, with little to no snags present. Water temperature cues for migration were found to be consistent with previous observations in other Gulf sturgeon rivers. Sedimentation, low dissolved oxygen concentrations, abandoned limb lines and a large logjam in the Escambia River were four major threats to the Conecuh/Escambia River Gulf sturgeon identified during this study.

CREVICE USE BY SALAMANDERS IN NORTHEASTERN ALABAMA WITH AN EMPHASIS ON THE GREEN SALAMANDER, ANEIDES AENEUS. Sara E. Viernum and George R. Cline, Dept. of Biology, Jacksonville State University, Jacksonville, AL 36265.

Northeastern Alabama supports a high diversity of salamanders, approximately twenty-eight species represented by five families (Ambystomidae, Crytobranchidae, Plethodontidae, Proteidae, and Salamandridae). These salamanders utilize terrestrial, aquatic, and fossorial habitats. Crevice use by terrestrial, Plethodontid salamanders was studied at two locations in DeKalb County; DeSoto State Park and Boy Scout Camp Comer. DeSoto State Park was relatively undisturbed, while both undisturbed and heavily disturbed sites were found at Camp Comer. Crevices in sandstone rocky outcrops located in mixed deciduous forests were surveyed bi-weekly beginning in May 2005. Height, width, and distance to ground for each inhabited crevice were recorded at each site. Five species of salamanders were collected: Aneides aeneus, Plethodon glutinosus, Eurycea longicauda, Pseudotriton ruber, and Notophthalmus viridescens. The most abundant species were A. aeneus, P. glutinosus, and E. longicauda. These three species appeared in the crevices about the same time in May. Both A. aeneus and P. glutinosus were observed regularly, while E. longicauda occurred sporadically. A. aeneus occupied shorter and narrower crevices, while P. glutinosus occupied crevices that were higher from the ground. Only A. aeneus was found at the disturbed sites of Camp Comer. A. aeneus densities appear higher in the disturbed sites than the undisturbed sites. Seasonal activities were also recorded for A. aeneus. Morphological reproductive structures, such as well-developed yellow mental hedonic glands on males, were noted in June. A brooding chamber containing approximately eight eggs was discovered in August. Hatchlings from the brooding chamber appeared in October.

MICROTUBULE INVOLVEMENT IN BALBIANI BODIES OF ACHETA DOMESTICUS OOCYTES. Thomas G. McFaden, Karen G. Wolfe, and James T. Bradley, Dept. of Biological Sciences, Auburn University, AL 36849.

Previtellogenic oocytes of the cricket Acheta domesticus contain an opaque cytoplasmic Balbiani body (BB) at each pole. BBs are visible in living tissue by light microscopy, and TEM shows them to contain dense aggregates of mitochondria. A single BB first appears at the proximal side of the germinal vesicle during early previtellogenic development. As this BB migrates to the proximal pole of the oocyte, a second BB appears near the distal side of the germinal vesicle and subsequently migrates distally. The incidence of BBs in ovaries was scored in last instar nymphs using differential interference contrast microscopy to determine the frequency of ovarioles containing visible BBs. To determine whether microtubules (MT) play roles in BB migration and/or stabilization, animals were injected with colchicine to give an in situ concentration of 0.2 mM. The depolymerizing effect of the colchicine solution on MTs was established using HeLa cells and flow cytometry to detect mitotic arrest. Results indicated that colchicine induces disappearance or dispersal of BBs inside oocytes of live crickets. We conclude that MTs are required for normal BB morphology and behavior. Work is underway to localize MTs inside oocytes using immofluorescent microsopy.

MICROARRAY ANALYSIS OF LEAD TREATED SACCHAROMYCES CEREVISIAE. In Ki Cho and Christi Magrath, Department of Biological and Environmental Science, Troy University, Troy, AL 36082.

The genome wide effect of lead acetate on gene expression in Saccharomyces cerevisiae was assessed using microarray technology. S.cerevisiae cultures were exposed to varying lead acetate concentrations and total RNA was prepared from each sample. Global gene expression profiles were generated by hybridizing labeled RNA to microarray (gene chips) obtained from the Genome Consortium for Active Teaching. Comparative analysis allowed identification of lead acetate responsive (LAR) genes with approximately 68 genes identified as responsive (48 repressed genes and 20 induced genes). Continuing investigation of these strains will allow elucidation of lead response mechanisms in S.cerevisiae.

MOLECULAR ANALYSES OF CADMIUM TOXICITY IN ARABIDOPSIS, SORGHUM, AND TOBACCO. Rachel J. Dunkerley and Mijitaba Hamissou. Department of Biology, Jacksonville State University, Jacksonville, AL 36265.

Cadmium is a non essential heavy metal pollutant of the environment, resulting from agricultural practices, mining, and industrial activities. Because of its high water solubility, Cd is readily available for uptake by plants and aquatic animals. In plants, Cd has been shown to interfere with the uptake, transport, and use of several essential elements causing complex nutrient deficiency symptoms. Several plant species are known to uptake and store Cd in their chloroplast, therefore affecting various aspects of photosynthesis. Like other heavy metals, the toxic actions of Cd are believed to be exerted on metabolic enzymes by altering their active sites. Cd toxicity in plants is also characterized by a rapid increase in reactive oxygen species (ROS) creating therefore conditions for secondary oxidative stress. The objectives of this research are to investigate the effects of Cd on the activities of ROS scavenger molecules, superoxide dismutase (SOD) and peroxidase (PO) enzymes and on the photosynthetic activities of isolated chloroplasts in Arabidopsis thaliana, Sorghum bicolor and Nicotiana tobacum challenged with elevated concentrations of Cd. Plants were grown in potted soil at pH 6.9 and fed with various concentrations of CdC12 for 14 days. Toxicity index chlorosis was recorded daily. Biomass accumulation, SOD, PO, and chloroplast activities were determined at the conclusion of the experiments.

RED TEARS: PORPHYRIN AND THE REX RAT. Elizabeth Swift, The Altamont School, Birmingham, Alabama 35213.

The purpose of this project was to test the hypothesis that porphyrin secreted by the Harderian gland in Rex rats (Rattus norvegicus) in the form of "red tears" has antibacterial effects on bacteria that grow on Rex rats. The rats were swabbed for bacteria, which were then allowed to grow on agar plates. The rats were then swabbed for porphyrin, which was applied to colonies of bacteria. The porphyrin had no noticeable antibacterial effects. However, the bacteria harvested from the parts of the rat with no porphyrin on them were markedly different than those harvested from areas that had porphyrin.

THE ONCOGENE N-MYC AND ITS REGULATORY ROLE IN THE TRANSCRIPTION OF N-ACETYLGLUCOSAMINYLTRANSFERASE VB IN NEUROBLASTOMAS. Marae M. Bernard and Jeanette Runquist, Birmingham-Southern Col., Birmingham, Al, 35254. Karen Abbott and Michael Pierce, Univ. of GA CCRC, Athens, GA 30602.

N-acetylglucosaminyltransferase VB (GnT-VB) is an enzyme that catalyzes the glycosylation of [beta] (1,6) branches on cell surface adhesion receptors. This glycosylation has been positively correlated with cell invasiveness. N-myc, an oncogene highly elevated in aggressive neuroblastomas, was theorized to have a potential link to the regulation of GnT-VB. To measure the effect of N-myc on GnT-VB expression, initial RT-PCR reactions were performed to measure basal levels of GnT-VB and N-myc in SK-N-SH, SH5Y, IMR32, and NBFL neuroblastoma cell lines. The levels of GnT-VB were positively correlated with N-myc levels in each cell line. Further investigation with RT-PCR revealed that the overexpression of N-myc in NBFL cell lines resulted in an 8-fold decrease in GnT-VB mRNA levels. Likewise, HEK cell lines transfected with an N-myc overexpression vector were detected to have decreased promoter activity. HEK cell lines transfected with the putative GnT-VB promoter mutated at the N-myc binding site were detected to have increased promoter activity, with wild type promoter activity at 2.92% of the positive control and mutant promoter activity at 29.6% of the positive control. In addition, when the N-myc overexpression vector was introduced in tandem with the mutant GnT-VB promoter, promoter activity decreased to 14% of the positive control. Overall, N-myc appears to negatively regulate GnT-VB activity, but further study is needed to clarify any distinct regulatory relationship.

SOME EFFECTS OF PHYLOXINE B ON TETRAHYMENA PYRIFORMIS IN VITRO. Misty Chapman, Mark Meade, Benji Blair and C. P. Olander, Dept. of Biology, Jacksonville State University, Jacksonville, AL 36265.

Phloxine B is a water-soluble halogenated xanthene dye that has recently gained popularity as an insect pesicide. Once ingested in baits, the dye is photoactivated causing tissue damage and the ultimate demise of the animal. Currently, researchers are examining the efficacy of phloxine B as and externally applied therapeutic bathe for fish infested with protozoan. In this study, we examined the effects of various concentrations of phloxine B on Tetrahymena pyriformis, a closely related protozoan to the Ichthyophthirius multifiliis a more pathogenic species that reeks havoc on fish farming. Acute exposure at 10mg/L phloxine B after 8hr in the light significantly reduces the number of cells in culture. Chronic exposure to lower concentration on cell growth, oxygen consumption, and phagocytosis are reported.
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Publication:Journal of the Alabama Academy of Science
Article Type:Conference notes
Geographic Code:1U600
Date:Apr 1, 2006
Words:11364
Previous Article:AAS Fall 2005 Executive Committee meeting.
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