Biological Sciences paper abstracts.

"DETECTION OF TOTAL AND PATHOGENIC SALMONELLA IN OYSTER USING REAL-TIME PCR" Maulshree Gangwar, Department of Biology, University of Alabama at Birmingham Dr.Asim K.Bej, Department of Biology, University of Alabama at Birmingham. Birmingham, AL 35294.

Salmonella is a causative agent for food and water borne gastroenteritis in humans. A multiplex PCR-based detection revealing total and potentially virulent strains of Salmonellae was developed using oligonucleotide primers. These primers targeted chromosomally located invA gene generating a 0.274 kbp and Salmonella virulence plasmid-borne spvB gene generating 0.561 kbp amplified DNA fragments. The invA gene product brings about the invasion of the cells on the intestinal epithelium and is used as the marker for total Salmonella detection. The occurrence and distribution of these genes in 90 strains exhibited 100% amplification of invA gene whereas only 22% of spvB gene implying their virulent nature. Real-Time PCR platforms, in contrast to conventional PCR, are rapid and obviate the need for further confirmation steps such as gel electropho-resis. The objective of the present study was to establish simple and specific methods to detect invA and spvB gene coding using SYBR-green Real-Time PCR. In each reaction, 16S rRNA gene was targeted as a competitive Internal Amplification Control (IAC) and used to identify detection of any false positives. The sensitivity of the reaction at all 3 targeted gene loci was found to be 1 pg. The analysis showed specific PCR products identified by melting curve analysis, and a reproducible melt temperature in the range of 88.0 to 89. 53[degrees]C (spvB), 83.06 to 85.48[degrees]C (invA) and 86.32 to 87.99[degrees]C (16S rRNA) was observed for all Salmonella strain. Negative controls and non-Salmonella strains showed an IAC-specific melt peak at 77.0[+or-]2.0 [degrees]C, giving evidence of the specificity of the method. The sensitivity of Real-Time PCR was found to be 1 ng that is comparable to approximately 104 cfu Salmonella enterica. The sensitivity was confirmed in pure cultures as well as in oyster tissue homogenate enriched with Salmonella.

AMYLOID [beta] 42-BINDING D-AMINO ACID PEPTIDE TREATMENTS AFFECT AMYLOID DEPOSITION AND REDUCE INFLAMMATION IN APP/PS1 MUTANT MICE Amy Nelson, Inga Kadisha and Thomas van Groen, University of Alabama at Birmingham, Birmingham, AL, 35294. Dieter Willbold, Forschungscentrum Juhlich, Juhlich, Germany.

Alzheimer's disease (AD) is an irreversible disorder of the brain causing loss of mental and physical function eventually leading to death. This disease is becoming more prevalent with an estimated thirteen million individuals predicted to be diagnosed by the year 2050. We are attempting to develop a better treatment modality for AD that improves cognitive function. To do so, we are using transgenic mice which express two AD mutations, PS1 [DELTA]E9 and APPswe. These mice develop three types of deposits: plaques, diffuse deposits, and cerebral amyloid angiopathy. These different amyloid deposits display distinct inflammatory characteristics. We hypothesized that long-term treatments that interact with A[beta]42 would result in changes in amyloid deposition and, likely, in the inflammatory reaction. Thus, we investigated the effects of small, A[beta]42-binding peptides that consist of D-amino acids on amyloid deposition. After two month treatment with amyloid [beta]42-binding peptide D3 there is a slight improvement in cognitive function, and a significant reduction in deposition of amyloid[beta]. A [beta]-load was significantly reduced in the cerebral cortex and hippocampus, and inflammation was significantly decreased around the amyloid deposits in the D3-treated mice compared to the control. Together these suggest that the amyloid-binding peptide influences the aggregation characteristics of amyloid[beta] and the inflammatory characteristics of the amyloid fibrils. Thanks to Drs. van Groen, Kadisha, Wyss and Watts for support and encouragement.

ANALYSIS OF SOUTHEASTERN HERPETOLOGICAL COMMUNITIES: SALAMANDERS. Brett Macek, George Cline, and Robert Carter. Biology Department, Jacksonville State University, Jacksonville, AL 36265-1602.

In this study, we examine the salamander communities of 26 sites in the southeastern United States. All species lists were taken from published manuscripts, or from unpublished projects with the approval of the researchers. Taxonomic changes that have been made since these papers were published have been corrected here when possible. Fifty-four species were recorded from these sites. Species richness (17 and 31 species). Ten sites had high species richness (8-13 species), but half" of the sites had low species richness (1-6 species). Most of the species had very narrow distributions; 26 species were only found at one of the sites and 47 species were found a 8 or fewer of the sites. Only 7 species were found at more than 8 sites (9-16 sites). This is similar to patterns seen in southeastern frogs, but salamanders appear to be more restricted to specific sits/habitats than frogs. Distribution patterns were analyzed using cluster analysis and principle components analysis correspondence analysis.

CAFFEINE AFFECTS EMBRYOLOGICAL DEVELOPMENT OF THE SEA URCHIN, LYTECHINUS VARIEGATUS. Alicia L. Kindred, Anita Patel, Victoria K. Gibbs, Addison L. Lawrence and Stephen A. Watts, Department of Biology, University of Alabama at Birmingham. Birmingham, AL 35294.

After the decoding of the S. purpuratus genome, sea urchins have become an important biomedical model for studies in toxicology and embryological development. Currently, research is limited using sea urchins as models of how xanthines affect embryonic development. Since the 1980 FDA warning, expectant mothers were advised to refrain from consuming caffeine, a commonly consumed xanthine. Caffeine (1, 3, 7 -trimethylxanthine) is a legally-used drug, heavily consumed daily by adults. Available data on caffeine shows that, in most mammals, caffeine serves as a mild teratogen. Using Lytechinus variegatus as a model, the effects of caffeine on embryological development were assessed. Toxicological tests on fluid-phase caffeine exposure were assessed using protocols established by the USGS. Sea urchins (30-40 mm diameter) were fed an experimental feed containing 0, 50, or 200 mg caffeine/kg feed for 16 weeks. Randomly selected urchins were spawned, and their F1 progeny were allowed to develop for 24 hours. Remaining urchins were dissected, and collected tissues were sectioned for histology. Caffeine had no adverse effects on weight gain, but toxicological tests indicate that caffeine significantly increased the rate of development (P < 0.05) for embryos whose parents were exposed to dietary caffeine. These data indicate that prenatal exposure to caffeine may have long-term effects on embryological development, the consequences of which are not known. Supported by Mississippi-Alabama Sea Grant Consortium.

CAUSE OF INJURY AND STATUS OF RAPTORS BROUGHT INTO THE REHABILITATION FACILITY OF THE MOBILE COUNTY ENVIRONMENTAL STUDIES CENTER IN MOBILE, ALABAMA. Shannon K. Markey, Dept. of Biology, University of South Alabama, Mobile, AL 36688.

Increased urbanization, habitat loss, and other ecological threats have caused many raptors to alter their habitat selections into more urban areas. Because of habitat alteration, many raptors are now encountering new environmental hazards. After the population decline that occurred in the mid-twentieth century and the recent recovery (Kirk and Hyslop, 1998). any new environmental hazards pose a significant threat to raptor populations. In this study raptor populations from the Alabama Gulf Coast have been monitored for fluctuations in mortality. Using local wildlife rehabilitation facilities, mortality and injuries have been documented. Many significant changes in the numbers of incoming raptors have been observed. Since 1993, there has been a significant increase in the number of Cooper's Hawks and a significant decrease in the number of Screech Owls admitted to the wildlife rehabilitation facility of the Mobile County Environmental Studies Center each year. With an occurrence rate of 29.7%, the most common injuries observed were those associated with neurological trauma. The goal of this study is to disclose major changes in population trends in the local raptor populations, as well as to determine the major environmental hazards faced by raptors in southwestern Alabama.

CHANGES IN ANTIBIOTIC RESISTANCE PROFILES OF HUMAN E. COLI: 2001-2004. Stacey E. Lawton and Brian S. Burnes, Department of Biology, Judson College, Marion, AL 36756.

Acquired bacterial resistance can be important in public health, raising husbandry, and tracking pollution. For this reason antibiotic resistance patterns of human E. coli, collected from the City of Marion wastewater treatment plant, were monitored through the years 2001 to 2004. The E.coli were isolated using ColiScan plates and profiled for resistance to a number of antibiotics. A total of 384 E. coli were tested for each year. Antibiotic resistance decreased for ampicillin by 60%, chorlamphenical by 57%, erythromycin by 43%, neomycin by 38%, oxytetracycline by 40 %, spectinomycin by 64%, streptomycin by 29%, tetracycline by 48%, and vancomycin by 7%. Vancomycin resistance decreased slightly, however E. coli would not be expected to be resistant to Vancomycin because it is a gram positive broad spectrum antibiotic. A total of 112 patterns were seen in 2001, 9 unique patterns were seen and a total of 29 patterns were also seen in 2004.

COMPUTATIONALAND BIOLOGICAL SCREENING ASSAYS FOR ENDOCRINE DISRUPTORS. Jonathan Catrett and Philip D. Reynolds, Department of Biological and Environmental Sciences, Troy University, Troy, AL 36082.

Endocrine disruptor chemicals (EDCs) are molecules that interact with and interrupt normal hormone-receptor signal transduction pathways. EDC exposure has been linked to detrimental changes in reproductive health and increases in reproductive diseases such as prostate cancer. Numerous chemicals ranging from pesticides to plasticizers have shown endocrine disrupting effects in both cellular and animal-based systems. Current experimental assays are accurate but are limited to studies on only a small number of molecules. With the use of potential EDCs growing rapidly in agricultural, industrial, and consumer products, the need for high-throughput screening is essential in identifying chemicals that are potentially harmful to the reproductive health of both humans and wildlife. In this report, a computational model was developed using pharmacological docking software to predict small molecule interactions with the androgen receptor, a hormone-receptor system that regulates male reproductive development. The hormone binding domain of the androgen receptor crystal structure was used with testosterone as the reference hormone. Several hormones, inhibitors and EDCs known for their ability to bind to the androgen receptor were tested in the computational model to see if the software could accurately predict their binding characteristics. The predictions were then compared to biological data using a new luciferase-based reporter gene assay in HeLa cells. The results indicate that receptor docking predictions may provide a primary screening tool to efficiently and accurately screen large numbers of chemicals for interactions with the androgen receptor.

Corynebacterium jeikeium Growth and Protein Expression When Studied in Serum. Rusty L. Yates, Nazia Mojib, and Asim K. Bej, Department of Biology, University of Alabama at Birmingham. Birmingham, AL 35294.

Corynebacterium jeikeium is a gram positive, opportunistic pathogen that could cause blood born infections including bacteremia, septicemia, and endocarditis; primarily in immunocompromised individuals having had open heart surgery. The growth response of this pathogen in medium with 10% (vol/vol) calf serum displayed a doubling of cell density within a 3 hour (h) period when compared to medium without serum. The growth response of C. jeikeium in 10% (vol/vol) human serum supplemented medium displayed an increase in cell density of more than two and a half times that of medium without serum after 2 h of growth. At 3 h of growth, C. jeikeium grown in medium with 10% (vol/vol) human serum had increased in cell density to almost four times the cell density of culture grown in medium without serum. An investigation of divalent cations that may be essential for growth in serum indicated[ Mg.sup.2] could be necessary for the cell density increase of C. jeikeium when grown in medium supplemented with serum. Following serum induced growth with medium supplemented with serum, total cellular proteins were radiolabeled and analyzed. Differential expression of at least two proteins, approximately 36-kDa (CSIP36) and 45-kDa (CSIP45), respectively, were observed. CSIP36 was found to be unique when compared to NCBI protein database after N-terminal microsequencing. The elevated expression of proteins of C. jeikeium when in serum is suggested by these results to possibly be necessary for a rapid increase in cell density during infection of a host's blood stream.

Creation of Neoplastically Transformed Human Mammary Epithelial Cells and Analysis of the Transcriptional Regulators of the human telomerase reverse transcriptase (hTERT) gene. Tyson DeAngelis, Yuanyuan Li, Anna Pendleton, Trygve Tollefsbol, University of Alabama at Birmingham. Birmingham, AL 35294.

Human mammary epithelial cells can be induced to undergo a transformation to a neoplastic phenotype, through the viral mediated introduction of three genes SV40 ER, hTERT, and an oncogenic form of hRAS, hRASv 12. This system provides us with the ability to study the very early stages of tumorigenesis in real-time. The human telomerase reverse transcriptase (hTERT) gene codes for the catalytic subunit of telomerase and its upregulation provides cancer cells with the ability to replicate indefinitely. The cause of the aberrant increase in hTERT expression in cancer cells has yet to be explained. Transformed HMECS (T-HMECs) that stably express all three genes display many, if not all of the characteristics of cancer cells, including the cancer-specific upregulation of endogenous hTERT. Careful analysis of the multiple transcription factors that have been shown to regulate hTERT has helped us to begin to elucidate the cause for the cancer specific upregulation of hTERT in breast cancer.

DETERMINATION OF SEDIMENT LOADING AT UNPAVED ROAD CROSSINGS IN SOUTHEASTERN ALABAMA. Suman Chitrakar, Neil Billington, and P. Michael Stewart, Department of Biological and Environmental Sciences, Troy University, Troy, AL 36082.

Unpaved road crossings have been a dominant source of sedimentation causing degradation of water quality and deterioration of aquatic habitat in southeastern Alabama streams. However, quantitative data on sediment loading at unpaved road crossings in this region are still limited. There is urgent need to quantify sediment delivery at unpaved road-stream crossings. Recently, a sediment risk index (SRI) for assigning risk of sediment loading at unpaved road-stream crossings has been developed at Troy University. This index will be used to identify 10 high risk and 10 low risk stream crossings, determine major variables that govern sediment production at these crossings, and quantify actual sediment delivery into the stream. Aluminum nails and rebar were used to gather data on sediment loading following rain events, along with rain gauges to determine precipitation. For a precipitation event of 20.32 mm, sediment loading ranged from 0.25-1.27 cm in a preliminary survey. Quantifying sedimentation will permit more focused and effective management practices to maintain stream water quality in the watershed.

Distribution and Habitat Characteristics of the Confederate Daisy, Viguiera porteri, In the Valley and Ridge Province of Alabama David M. Frings, Department of Biology, Samford University, Birmingham, AL 35229.

Viguiera porteri is documented in the literature as growing in xeric habitats on granitic outcrops in Georgia and extreme eastern Alabama. Local botanists have known that a population of V. porteri occurs in a similar xeric habitat founded on sandstone at Oak Mountain State Park. The extension of the range of the species to the Valley and Ridge Province of central Alabama has not been established in the literature. The xeric outcrops that support the population of V. porteri at Oak Mountain State Park are formed on Pennsylvanian age, sandstone outcrops. This investigation compares the characteristics of the habitat of V. porteri at Stone Mountain, Georgia to those found at Oak Mountain State Park in order to determine the habitat conditions that are required to support the population. Parameters that were investigated include the rock type, soil characteristics, soil pH, elevation, and exposure to sunshine. Since V. porteri is restricted to xeric outcrops, topographic maps were used to locate additional areas that contain similar habitats and may support new populations. Two areas investigated were Moss Rock Preserve on Shades Mountain in Hoover, Alabama and Bald Rock Mountain in St. Clair County, Alabama. A new population of V. porteri was located at Bald Rock Mountain in St. Clair County, Alabama. No population was found at the Moss Rock Preserve in Jefferson County, Alabama. The field investigation shows that V. portersi requires a xeric habitat with a minimal amount of soil. Soils encountered were either derived from the weathering of granite or sandstone, were very sandy, and had an acidic pH. Plant populations were robust in areas that had a southern exposure to full sunshine. No plants were observed at elevations lower than 900 feet above mean sea level which may be the reason that V. porteri was not observed at the Moss Rock Preserve.

EFFECTS OF DIETARY ZINC ON GROWTH AND REPRODUCTION OF THE SEA URCHIN, LYTECUINUS VARIEGATUS. Kimberly N. Trawick, Jessica E. Etling, Addison L. Lawrence, Stephen A. Watts, Department of Biology, University of Alabama at Birmingham, Birmingham, AL 35294.

Development of sea urchin feeds for sea urchin aquaculture requires evaluation of proximate constituents as well as other important macro- and micronutrients. Despite the production of a large calcite test, mineral requirements have not been determined for any species. This study examined the effects of dietary zinc on organismal and organ weight gain. Adult sea urchins (26.6 g wet wt.) were separated randomly into different treatment groups (n = 18 individuals per treatment), and were fed one of seven formulated feeds that were supplemented with different levels of dietary zinc (4.5, 10.75, 26.32, 65, 161.7, 403.2, 1005 ppm, final concentration). After nine weeks, the sea urchins were dissected and wet and dry weights of the organs were determined. Dietary zinc did not significantly affect the test or gut growth. The formulated feed without zinc supplementation resulted in reduced gonad weight gain. Moisture content was reduced in those individuals, suggesting a difference in proximate composition. These data indicate dietary zinc is required for gonad production. Sea urchins fed 1005 ppm zinc had a significantly smaller lantern, which suggests high levels of dietary zinc are toxic. Supported in part by Mississippi-Alabama Sea Grant Consortium.

Evaluating Androgenic Steroids using Xiphophorus helleri. Robert R Parrish, Joesph M Hales, Jon Hopper, and Tomas E. Denton, Department of Biology, Auburn University Montgomery, Montgomery, AL 36124.

Male swordtail fish (genus Xiphophorus) possess a long, colored extension of the caudal fin. This trait, seen only in the male fish, is an example of sexual selection. Immature males are swordless but develop the sword extension during the sexual maturation process. During maturation, the male swordtail fish have an increase in testosterone levels, which leads to the development of the caudal fin extension, commonly referred to as the sword. Although this trait is not seen in normal female swordtail fish, many previous studies have reported female swordtails being transformed into phenotypic male fish by the administration of male sex hormones. In this study, female swordtails were exposed to four different C-19 steroids over a fourteen-day period: two adrenal steroids and two testosterones. All four of the drugs produced caudal fin transformation with no sign of regression long after exposure to the drugs ceased. The effectiveness of the different steroids was measured against each other and a control. The steroids containing double bonds in their A rings typically appear to be more potent, causing greater numbers of caudal fin transformation than the steroids with a single bond in their A rings. One synthetic steroid, known as delta 1-adrenosterone, was tested and found to be equally as potent as delta 1-testosterone in caudal fin transformation. This delta 1-adrenosterone is unreported in any living species, and is therefore considered newly introduced into a living species.

FACTORS LEADING TO CANNIBALISM IN LAB-REARED LYTECHINUS VARIEGATUS (ECHINODERMATA). Cristina M. Richardsonand Stephen A. Watts, Department of Biology, University of Alabama at Birmingham, Birmingham, AL 35294.

Sea urchins are prized for their gonads, or uni, that are eaten as sushi in Asian and Mediterranean countries, as well as for their use as an important biomedical model for research. Their successful aquaculture has become vitally important in recent years as natural populations have been depleted by overfishing. Recent observations indicate that. when held in captivity en masse, adult L. variegatus will cannibalize other L. variegatus in the tank. Individuals consumed showed no signs of injury or disease. Cannibalism occurred when a predatory urchin first consumed the spines of a conspecific prey urchin, usually on the aboral surface, and then proceeded to break the test and consume completely the test and other organs. Cannibalistic behavior has recently been observed in dense populations of developing juveniles. In this study, juvenile L. variegatus ( < 2 mm diameter) were placed in small finger bowls and observed for up to two weeks. Individuals were not fed during the two-week period. Previously-starved juveniles showed minimal cannibalism when held at densities of either 1600/[m.sup.2] or 1000/[m.sup.2] (< 7%). Cannibalism increased to 23% in juveniles fed previously and then transferred to finger bowls for two weeks at 1600/[m.sup.2]. We suggest that nutritional history, size, and density will contribute to the rate of cannibalistic behavior in cultured sea urchins. This is the first report of conspecific cannibalism in any sea urchin species.

GENETIC VARIATION IN WALLEYE POPULATIONS. Sonia D. Lyle and Neil Billington, Department of Biological and Environmental Sciences, Troy University, Troy, AL 36082.

Walleye (Sander vitreus) (Percidae) is a large predaceous fish species that is common in Canada, and the mid-west and Great Plains region of the U.S. These fish are intensively managed because they are popular with anglers. Thus, information on the population genetic structure of walleye will be useful in their management. Cellulose acetate gel electrophoresis was used to screen genetic variation in 1188 walleye from nine populations. Two polymorphic protein-coding loci in walleye, malate dehydrogenase (sMDH-3) and general muscle protein (PROT-3), were surveyed. Five populations showed significant deviations from Hardy-Weinberg expectations at sMDH-3 and three populations showed significant deviations from Hardy-Weinberg expectations at PROT-3, all due to heterozygote deficits. This was likely caused by the Wahlund effect because most samples were collected during the summer and fall, a period when walleye are highly mobile, rather than during the spring when they are presumed to segregate into discrete spawning aggregates. Highly significant among population heterogeneity was found for walleye at both sMDH-3 and PROT-3, suggesting a high degree of differentiation among walleye populations. Managers are encouraged to manage walleye populations that are genetically distinct separately, because these populations may exhibit local adaptations. Mixing such populations might lead to outbreeding depression.

HYBRIDIZATION BETWEEN WALLEYE AND SAUGER. Rachael N. Koigi, Janet Gaston, Neil Billington, Ronald E. Creech, Amy M. Wotawa, and Sonia D. Lyle. Department of Biological and Environmental Sciences, Troy University, Troy, AL 36082.

Walleye (Sander vitreus) and sauger (S. canadensis) (Percidae) are both large predatory fishes that hybridize naturally and their [F.sub.1] hybrids backcross with both parental species leading to introgression. We examined 5385 fish collected from 15 populations in the Great Plains (8 populations) and the mid-west (7 populations) where both species co-occur. Four diagnostic protein-coding loci (ALAT and IDDH from liver, and mMDH-1 and PGM-1 from muscle) between the two species were screened by cellulose acetate gel electrophoresis to determine the proportion of hybrids (fish possessing alleles of the other species) in each population. For the Great Plains populations, no hybrids were found in two Wyoming populations, hybrids ranged from 2.8-11% in five Montana populations and 2.7-10% in potential Montana sauger brood stock, while in Lake Diefenbaker, SK, Canada, 17.1% of fish were hybrids. For the mid-west populations, 20.4% of sauger from Lake Sakakawea, ND, contained walleye alleles, whereas in South Dakota 4.2% of fish from Lake Sharpe, 3.9% from Lake Francis Case, and 21% from Lewis and Clark Lake were hybrids. In addition, 22.7% of fish from the Mississippi River Pool 13 (IA) were hybrids, 30% of fish from Ombabika Bay, Lake Nipigon, ON, Canada, and 4.1% of fish from the Illinois River and 1.9-4.1% of potential sauger brood stock fish were hybrids. Managers need to take hybridization into account when selecting brood stock from these water bodies.

MASCULINIZING POTENCIES OF METHYLTESTOSTERONE AND SPIRONOLACTONE IN FEMALE WESTERN MOSQUITOFISH, GAMBUSIA AFFINIS. Samiksha Raut, Robert Angus, Department of Biology, University of Alabama at Birmingham, Birmingham, AL 35294.

There is a growing concern regarding presence of anthropogenic compounds in the environment that can act as endocrine disruptors. Among such endocrine disrupting compounds, estrogens have been extensively studied. Although reports of androgenic endocrine disruptors continue to increase, this class of compounds has not been well studied. We are currently investigating the effects of the potent synthetic androgen methyltestosterone (MT) and aldosterone inhibitor spironolactone (SPL) on an androgen-dependent secondary sex characteristic (gonopodium) in female western mosquitofish, Gambusia affinis. Females were exposed for five weeks to MT (10-10000ng/L) and SPL (10-500nM) by using the static renewal method. Morphological masculinization as evidenced by the development of an elongated and modified anal fin was observed in the three highest exposure groups in both MT and SPL treated females. However, in the MT treated females, the anal fin ray elongation was completed by the second week of exposure along with the development of hooks and spines. On the other hand, SPL treatment resulted in increasing the anal fin ray length until the second week of exposure and showed the development of hooks and spines after three weeks. These results indicate that the potent masculinzing agent MT causes masculinization more rapidly than SPL. Further these results also confirm earlier studies indicating the androgenic mode of action of SPL in female mosquitofish in contrast to its anti-androgenic effects observed in humans. This work supported by AAS student research grant 2007.

Mesofauna Associated with the Subtropical/Tropical Marine Sponge Amphimedon viridis: Are Faunal Associations Likely to Provide Refuge from Fish Predation Due to Physical or Chemical Feeding Deterrent Properties of this Sponge? Jonathan Huang, James B. McClintock, Charles D. Amsler and Yusheng Huang, Department of Biology, University of Alabama at Birmingham, Birmingham, AL 35294.

Sessile benthic marine organisms such as sponges are frequently colonized by a wide diversity of invertebrates. The present study focused on a quantitative analysis of the mesofauna associated with the common subtropical/tropical sponge Amphimedon viridis and explored several criteria for the basis for this mesofauna-sponge relationship. Specimens of Amphimedon viridis were hand-collected individually in bags from both Saint Joseph Bay, Florida Panhandle, and Sugar Loaf Key, Florida Keys between April-November 2006. Quantitative analyses of sponge surfaces and interiors revealed that the most common sponge-associated group was comprised of amphipods, followed by polychaetes and tenaid crustaceans. 35 percent of sponge-associated individuals were recovered from the interstices of the sponge. A total of 707 individuals per 1 c[m.sup.3] of sponge material were found within 15 sponge samples of A. viridis. Shannon diversity and evenness indices indicated that A. viridis supports a diverse community of sponge-associated invertebrates including amphipods, polychaetes, tenaids, decapods, and isopods. These sponge-associated invertebrate groups may occupy sponges for a number of reasons. For example, physical or chemical protection from fish predation may impose a strong selective pressure that sustains these relationships. Moreover, nutritional needs, reproductive behaviors, and social interactions could also contribute to these sponge associations. In order to evaluate whether Amphimedon viridis provides a refuge from predation for sponge-associated mesofauna, laboratory feeding experiments were conducted to assess the palability of A. viridis to the generalistic sympatric pinfish Lagodon rhomboides. In feeding assays employing small pieces of sponge tissue and control squid tissue, pinfish displayed a strong significant rejection for sponge tissues when compared with control squid tissues. Alginate food pellets loaded with ecologically relevant concentrations of spicules isolated from A. viridis caused a weak but significant deterrent response in pinfish when compared to food pellets lacking spicules. However, alginate food pellets containing tissue-level concentrations of lipophilic and hydrophilic sponge extracts were highly deterrent to pinfish when compared to control food pellets. It is concluded that while both physical and chemical characteristics of A. viridis may contribute to its quality as a habitat resource, potent secondary metabolities etabolites, likely halotoxins and amphitoxins, play a particularly important role in providing a chemical refuge for associated mesofauana.

MICROBIAL LOAD OF SELECTED COLLEGE PROFESSORS. Diane M. Jowers, Tia McClenney, Genevieve Russell, and Brian Burnes, Biology Department, Judson College, Marion, AL 36756.

Samples from the mouths of selected faculty and students of Judson College were collected and identified to compare the microbial loads with respect to age, gender, smokers vs. nonsmokers, academic compartments, illness, hand washing habits, oral hygiene habits, contact with children, and gum chewing. Mouth swabs were taken and plated on Nutrient Agar and incubated at 37 degrees Celsius. The bacterial colonies were observed for numbers, morphology, gram stain, and Enterotube II results. Plate counts ranged from I to 2048 bacterial colonies, morphology included cocci and bacillus, and gram stains indicated both positive and negative bacteria. In conclusion, higher numbers of Gram positive cocci were present in members of the Biology department than members of the Education, Business, and Student Life departments, higher numbers of Gram negative cocci and Gram positive Bacillus were present in Education department members than Biology, Business and Student Life members, higher microbial loads were present in nonsmokers than smokers, and a higher number of males presented Gram positive spreading colonies than females. There was no direct correlation between hand washing habits and oral bacteria.

MITOCHONDRIAL DNA MARKERS FOR IDENTIFYING SOUTHERN AND NORTHERN STRAINS OF ALABAMA WALLEYE. Amy M. Wotawa and Neil Billington, Department of Biological and Environmental Sciences, Troy University, Troy, AL 36082.

Walleye (Sander vitreus) (Percidae) from the Mobile drainage of Alabama are rare, but genetically distinct from Tennessee River walleye populations. Early work based upon whole-molecule mitochondrial DNA (mtDNA) analysis showed that the northern strain and the southern strain (sometimes called the Gulf Coast strain) are separated by a sequence divergence of 2.36% meaning that they likely diverged approximately 1 million years ago. Because numbers of the southern walleye are declining we sought to find a non-lethal way of being able to determine differences between northern and southern walleye mtDNA in Alabama. A search of published D-loop sequence data from both strains revealed three restriction site changes in this region that could be revealed by PCR-RFLP analysis with four-base recognition endonucleases. Two site gains were detected in the southern strain compared to the northern strain for Rsa I and a site loss in the southern strain compared to the northern strain was found for Tru9 I. We intend to use these markers to identify northern and southern walleyes in Alabama by PCR-RFLP analysis. This should allow females of the southern walleye strain to be confirmed by genetic analysis prior to them being used in conservation breeding programs.

NECROPSIES OF CETACEANS AND THE COURTS. Gerald T. Regan 4000 Dauphin Street, Mobile, AL 36608-1791.

The National Marine Fisheries Service administers the Marine Mammal Stranding Network. For 20 years there has been a participant from A labama in that network. An important part of that participation has been the preparation of specimens and data that would be recognized in court cases involving allegations of violations of the Marine Mammal Protection Act of 1972 as amended. Alabama participants studied books on that subject and participated in half a dozen in-service workshops. Four hundred and forty-three dead or dying dolphins, whales, and manatees came under the scrutiny of Alabama participants. Most of them underwent necropsy. But no data or specimens were used by any court in arriving at a decision. That raised the question of what had happened in other states. A pilot study of the past 20 years of such records as are available to Academic LexisNexis showed that only one case had been decided on the basis of information from necropsies. Beaked whales had died in the Gulf of California after airguns had been used by Columbia University. The magistrate of a California district court ruled that there would be a temporary injunction against the use of the airguns based on necropsy data from several different species of cetaceans that died in association with the Navy's use of SONAR near the Bahamas. The necropsies showed that there had been irreparable harm to the ears and lungs. Spring Hill College, the Marterra Foundation, and the Prescott grant program assisted work mentioned in the report above.

OBSERVATIONS OF GROWTH IN NEWLY-METAMORPHOSED JUVENILE LYTECHINUS VARIEGATUS (ECHINODERMATA: ECHINOIDEA). Jessica E. Etling, M.L. Powell, A.L. Lawrence and S.A. Watts. Department of Biology, University of Alabama at Birmingham, Birmingham AL 35294.

Sea urchins are excellent biomedical and toxicological models, as well as a popular food among many people. However, sea urchin populations are being decimated by overfishing. Due to this high demand, there is an increasing need to develop methods for inland rearing of sea urchins from the larval to the adult stage. Sea urchin larvae transition from free swimming larvae feeding on unicellular algae in the water column to a benthic environment. Little is known about juvenile nutritional requirements at this early developmental stage. Feeding in newly metamorphosed juveniles was examined in settled sibling larvae collected from a single male and female spawn and placed in duplicate 8 L aquaria. Newly-metamorphosed urchins were placed in one of four treatments: Biofilms of the live diatom Amphora helenensis, A. helenensis plus diatomaceous earth, A. helenensis with a diatom substitute (ReedMariculture Inc.), and the diatom substitute alone. Individual growth was highly variable within all groups, with a range in diameter from 0.4mm to 3.0mm. Survival was comparable among the first three treatment groups, however, no individuals survived when fed diatom substitute alone. Supported in part by Mississippi-Alabama Sea Grant Consortium.

OPTIMIZATION OF A DISCRIMINANT FUNCTION FOR MICROBIAL SOURCE TRACKING. Brian S. Burnes, Department of Biology, Judson College, Marion, AL36756.

A potentially inexpensive method for determining the sources of pathogenic microbial contamination of surface and drinking water is to apply discriminant function analysis to antibiotic-resistance patterns. E.coli from Humans, Cattle, or Deer were collected, isolated, and tested for growth in the presence of various antibiotics (ampicillin, chloramphenicol, erythromycin, neomycin, oxytetracycline, spectinomycin dihydrochloride, streptomycin sulfate, and tetracycline hydrochloride) and these results were used to develop discriminant functions that correctly categorized E.coli of unknown origin with high accuracy. A discriminant function that classified E.coli into all 3 groups had an average rate of correct classification of 78%, whereas a discriminant function that classified E.coli into just two groups, human or non human, had an average rate of correct classification of 90%.

PACIFIC SANDS: THE QUEST FOR LIFE. Karl Fernandez, Angela Smith and George Williams, Athens State University, Athens, AL 35611.

Sands are deposited around the world on oceanic beaches, deserts, dunes, and sand bars of riverine environments. Transport media for sands include water, wind and ice. Sand is a vital component in marine ecosystems. Sands are formed from many sources including fast flowing rivers, mountains, volcanoes and several types of organic materials. We are especially interested in beach sands and those that contain tests, shells, and skeletal fragments of living organisms. We carefully examined several sands from the Pacific, with a few others for comparison, to determine characteristics of each and the approximate fraction and types of organic materials within the selected specimens. Sands from the Hawaiian Islands, California, Fiji, Kwajelein, the Marshall Islands, Guam, Pago Pago, Utah, and the Galapagos Islands were included in this study. Samples were carefully studied microscopically and photographed for colors, surface texture and degree of grain sorting. Sand grains were measured to determine size ranges, weighed, and tested for organics and magnetite. We found sands from Hawaii, Kwajelein, Fiji, the Marshall Islands, Guam, Pago Pago, and the Galapagos Islands are almost one hundred percent organic in origin. They include grains and fragments from corals, foraminiferan tests, gastropod shells, bivalve shells, sponge spicules, fossiliferous fragments, crinoids, sea urchin spines, and others.

Preservation of a Mature Canopy Floodplain Hardwood Forest: Perry Lakes Park and Environs Lauren Stephens *, * Angi Gullard *, * Tonya Morgan *, * Riki Enzor Judson College Earth Team - Dr. Thomas Wilson, Leader. Judson College, Marion, AL 36756.

Preservation of a Mature Canopy Floodplain Hardwood Forest: Perry Lakes Park and Environs Perry Lakes Park and associated Barton's Beach and the Marion State Fish Hatchery woods make up 900 acres of Cahaba River mature canopy floodplain forest. This rare and valuable bottomland ecosystem is being developed into a nature preserve, outdoor laboratory, research center, recreational area, and ecotourism attraction. This area represents a major birding destination second in importance only to Dauphin Island. The Park's 100-foot tall canopy tower allows a new approach to birding and to the study of trees. The wet areas are home to Swamp and Water Tupelos, Bald Cypress, Blackgum and Overcup Oaks. The numerous mature and dying trees create a prime habitat for large numbers of woodpeckers, especially the Pileated Woodpecker. The birding list for the Park is over 207 species. The Park was created in 1999 by a 20 year lease from the Alabama Department of Conservation and Natural Resources. There is a strong movement by Friends of Perry Lakes Park to have the Department of Conservation develope a management plan that designates the Park and Marion Fish Hatchery woods as a State Natural Area similar to a Federal wilderness. It is important that this wonderful and threatened ecosystem be preserved in its natural state for the well being of wild creatures including beautiful trees and plants. It is important for all of us that we realize that we can save and preserve some parts of nature for future generations. We thank Dr. Thomas Wilson, Judson College Biology Professor for leading the Earth Team in this project.

Spectacular Specimens in a Speck of Sand Angela Smith, Karl Fernandez, George Williams Athens State University, Athens, AL 35611.

Sand is a substance on which many do not dwell. When you think of sand, you think of some dirt-like substance found at the beach or in a desert. Not much thought is given toward the composition and make-up of sand, or to the organic material that can be found in sand. The purpose of this study was to examine the life forms found in sands from around the Atlantic Ocean. Ten sands were chosen for the study and comparison. We carefully examined several sands from the Atlantic, to determine the characteristics of each and the approximate fraction and types of organic materials within the selected specimens. Samples were carefully studied microscopically and photographed for colors, surface texture and degree of grain sorting. Sand grains were measured to determine size ranges, weighed, and tested for organics and magnetite. This study shows how sands from various points around the Atlantic can be so similar and so different. The sands used in this study were from Connecticut, Florida, the Bahamas, Ireland, Spain, France, Greece, and Africa. Results showed that the majority of the sands studied did contain some life forms. In just a speck of sand, so many specimens can exist. The life forms ranged from coral fragments, to mollusk shells, sea urchin spines, sponge spicules, gastropod shells, and foraminiferan tests.

STATUS OF THE SANDHILL CRANE, SNOW GOOSE AND CANADA GOOSE ON WHEELER NATIONAL WILDLIFE REFUGE, DECATUR, AL. Todd Cook and George Williams, Athens State University, Athens, AL 35611.

Wheeler National Wildlife Refuge has been a haven for migratory and resident birds since its establishment in 1938. The refuge staff maintains careful records of bird populations. Species numbers are determined each winter by actual ground counts and aerial surveys. We studied these data to determine the changes in numbers for the sandhill crane, Canada goose and snow goose. The sandhill crane surveys verify a steady increase in numbers at Wheeler from twenty-six in 1997, to a peak of 2,564 in January of 2008. Numbers of the Canada goose indicate a decline from a high of 58,000 in the winter of 1963-1964, to a low of 1,153 in the 2006-2007 season. The snow goose numbers range from a low of 100 in 1948-1949, to 2,800 in the winter of 1999-2000. The steady rise in the numbers of the sandhill crane may be due to their preference to winter farther north in recent years. The Canada goose numbers have declined possibly due to a combination of the loss of habitat in Southern James Bay, predation, hunting, and short-stopping to areas north of Wheeler. The snow goose population at Wheeler each winter has been the most consistent of the three species.

STEROID HORMONE LEVELS AND THEIR IMPLICATIONS FOR OVERALL REPRODUCTIVE PHYSIOLOGY IN THE DIAMONDBACK TERRAPIN, MALACLEMYS TERRAPIN. Andrew T. Coleman, Thane Wibbels, Ken Marion, Univ. of Ala. at Birmingham, Birmingham, AL 35294. Willem Roosenburg, Ohio Univ., Athens, OH 45701. David Nelson, Joel Borden, and Gabe Langford, University of South Alabama, Mobile, AL 36688.

Previous studies that examined reproductive steroid levels in other turtle species have contributed to the elucidation of certain physiological and ecological aspects such as reproductive activity, stress levels, and sex ratios. The diamondback terrapin, Malaclemys terrapin, is the only turtle in North America that exclusively inhabits the brackish environments of bays and estuaries. This species faces numerous threats throughout its range, so it is imperative to have a thorough understanding of factors relating to its reproductive physiology. In the current study, blood samples were obtained from two geographically separated populations, both of which are believed to be in decline. Testosterone and estrogen levels were measured via radioimmunoassay (RIA), and the seasonal cycles of these reproductive steroids were examined. Adult females showed a gradual decrease in testosterone over the nesting season, while adult males showed a gradual increase. No significant differences were observed in estrogen levels between gravid and non-gravid females. The results provide insights, which will be discussed, into the endocrine physiology underlying the reproductive cycle in the terrapin. This research was funded through grants from the Alabama Department of Conservation and Natural Resources and the Alabama Academy of Science.

STUDY OF DIFFERENTIALLY EXPRESSED PROTEINS IN ANTARCTIC BACTERIA AT COLD TEMPERATURE. Nazia Mojib and Asim K. Bej, Department of Biology, University of Alabama at Birmingham, Birmingham, AL 35294.

Several factors are involved in the mechanism of bacterial adaptation to low temperatures and one of them includes the production of Ice Binding Protein (IBP) which is well documented in snow mold and sea ice diatoms. In our study, various bacterial species from Antarctica, which survive at subzero temperatures were screened for the expression of IBP. Since IBP (25 kDa) from sea ice diatom Navicula glaciei show considerable similarity to psychrotolerant sediment bacteria, Rhodoferax ferrireducens (58% identity) and a denitrifying estuarine bacteria, Shewanella denitrificans (43% identity), polyclonal antibody raised against a conserved middle domain of IBP was used for detection of IBP in Antarctic bacteria. Out of 11 different microorganisms screened, 7 exhibited a single protein band ranging between 20 kDa and 30 kDa. Interestingly, IBP-like protein was also identified in the selected Antarctic pseudomonads which have previously been reported to produce ice-active and cold acclimation CapB protein. Degenerate PCR strategy was used to amplify IBP from Pseudomonas 30-3. There was marked elevation in the expression of IBP in Pseudomonas 30-3 at low temperatures (0[degrees]C and -5[degrees]C). This suggests that IBP may be one of the factors contributing to survival mechanism at low temperatures. Microscopic examinations to confirm the viability of these bacterial cells both at low and high temperatures using Molecular Probes Live/Dead Bac-Lite fluorescent stain exhibited live and actively growing bacteria at all stages. Further investigation of the structure and function of IBP will help understand the mechanism of adaptation of Antarctic bacteria at sub zero temperatures.

SURVEY OF AEROSPORA AT DIFFERENT HEIGHTS: QUALITATIVE AND QUANTITATIVE ASSESSMENT. Marian Wehby and H. Wayne Shew, Birmingham-Southern College, Birmingham, AL 35254.

Airborne pollen and fungal spores are a leading cause of allergies. Air samples collected at certified pollen stations are used to count numbers and types of pollen and fungal spores, and these counts are used by allergists to predict the greatest risk for pollinosis in individuals due to the presence of a particular aeroallergen. Currently, official, reported pollen counts are determined using pollen collectors located on roof tops (at a height of 12-20 m). Studies in Spain, Italy, and Northern Europe have shown significant differences in the amount of pollen present at different heights, but a similar study has not been performed in the United States. This study was undertaken to determine whether air samples taken at ground level, eye level (1.5 m), and roof level (13 m) show significant differences in the concentration and types of pollen and fungal spores. Samples were collected on silicone coated microscope slides during October, November, and December 2007, and January 2008, using a Burkard volumetric air sampler. Samples were mounted in glycerin jelly containing phenol and basic fuchsin, and examined with light microscopy. The results were statistically analyzed using an unpaired two-tailed t-test. Preliminary results indicate that there are significant differences in types and number of fungal spores per [m.sup.3] at different heights. Very low pollen counts during the months in which samples were taken prevent any definitive statement about the effect of sampling heights on pollen types and number per [m.sup.3].

THE EFFECT OF DIETARY CALCIUM ON ZEBRAFISH (DANIO RERIO) GROWTH, SURVIVAL, AND BONE MINERAL DENSITY. Steve Padgett-Vasquez, Anthony J. Siccardi III, Lou R. D' Abramo, Stephen A. Watts, Department of Biology, University of Alabama at Birmingham, Birmingham, AL 35294.

Osteoporosis affects one third of women over the age of 65. It has been studied extensively in human and animal models. Zebrafish (Danio rerio) may be an excellent bone model because of lamellar structure and hierarchical organization (levels one through seven) similar to human long bones. Calcium is the most abundant mineral in the human body. It is present in both intra and extracellular fluids, but it is most abundant in the bone. Juvenile, reproductively-intact zebrafish from day 28 through 112 post-fertilization were used in two separate scenarios due to their natural ability to sequester calcium through their gills. One group was placed in RO filtered water lacking calcium (approximately 0 ppm) and were fed 0, 2, 4, 6, or 7.7 mg/g feed of calcium carbonate. The second group used filtered tap water that contained calcium in the water (approximately 25 ppm) and were fed 0, 2, 4, or 6 mg/g feed of calcium carbonate. After the twelve week study there were no significant differences in weight, length, survivorship, or bone mineral density (determined by micro CT) of any treatment groups, regardless of calcium in the water.

WHOLE-CELL SACCHARIFICATION OF LIGNOCELLULOSES, Benedict C. Okeke, Department of Biology, Auburn University Montgomery, Montgomery, AL 36124.

Fossil fuel is expensive and nonrenewable. Inedible plant materials such as wood chips, wood shavings and grass are abundant renewable natural resources that can be transformed to biomass fuel. Plant materials are composed largely of lignocelluloses which are complex polymers of cellulose, hemicelluloses and lignin. Cellulose, a linear glucose polymer, is the principal structural polysaccharide in plant fiber. Hemicelluloses are heteropolymers of a range of pentose sugars, primarily xylose and arabinose. Cell-free enzymes have been widely studied for lignocellulose saccharification. A major limitation of cellulose breakdown to fermentable sugars is incomplete mixture of enzymes and instability of extracted enzymes. Use of whole-microbial-cells has the advantage of secretion of a wide spectrum of synergistic enzymes required for complete degradation of lignocelluloses. This paper presents the development of a process for whole-cell saccharification of lignocelluloses for production of biomass fuel.

ROAD-KILL SURVEY OF ALABAMA RED-BELLIED TURTLES ON THE MOBILE BAY CAUSEWAY. David H. Nelson and Cynthia Scardamalia-Nelson, Department of Biological Sciences, University of South Alabama, and Providence Hospital, Mobile, AL 36688.

A systematic, road-kill survey was conducted (by bicycle or automobile) on the Mobile Bay Causeway from April 2001 to December 2007 to assess the numbers of Alabama red-bellied turtles (Pseudemys alabamensis) killed by automobile traffic. A federally endangered species, Pseudemys alabamensis has been designated as the official "Alabama state reptile." A total of 553 Alabama red-bellied turtles were recorded over the seven-year study: 420 hatchlings, 116 adult females (most gravid), 13 juveniles, and 4 males. A majority of hatchlings (96%) over-wintered in the nests to emerge during the following Spring (March-May). Fewer numbers of hatchlings (4%) emerged during the Fall (October and November) of the same year. Direct hits by hurricanes apparently resulted in fewer roadside mortalities of hatchlings (as they were drowned or emerged prematurely). The mortality of adult females (N=116 was greatest (92%) during the nesting season: May, June, July. Each year, from 5 to 34 nesting females, mostly gravid (mean = 16.6), were killed by vehicular traffic on the road. Because of the limited availability of favorable nesting sites in the lower delta, gravid females are apparently attracted to the shoulders of elevated roadsides where they deposit eggs (and may incur mortality). A chain-link fence is currently being installed by the Alabama Department of Transportation to reduce the road-side mortality of turtles along the Mobile Bay causeway. Partial funding was provided by the Alabama Department of Conservation & Natural Resources and the U.S. Fish and Wildlife Service.

Phototactic Responses of Larvae from the Marine Sponge Xestospongia proxima. Andrew S. Mobley, Department of Biology, University of Alabama at Birmingham, USA. Linnet Busutil Lopez, Instituto de Oceanologia, Cuba. Sally P. Leys, University of Alberta, Canada. Maria Cristina Diaz, Museo Marino de Margarita, Venezuela. Rachel Collin, Smithsonian Tropical Research Institute, Bocas del Toro Research Station, Panama. Robert W. Thacker, Department of Biology, University of Alabama at Birmingham, AL 35294.

In August 2007, the Training in Tropical Taxonomy program at the Smithsonian Tropical Research Institute's Bocas del Toro Research Station sponsored a two-week course on the Taxonomy and Ecology of Caribbean Sponges. During the course we examined the phototactic behavior of larvae from the marine sponge Xestospongia proxima. Previous studies suggest that negative phototaxis in sponge larvae is generated by the responsiveness of a tuft of long posterior cilia. While other sponge larvae are known to be positively phototactic, the mechanism by which swimming direction is coordinated in those species is not yet understood. Since X. proxima hosts large populations of photosynthetic, unicellular cyanobacteria, we hypothesized that the larvae would be positively phototactic, and that the ciliary response would be opposite to that of negatively phototactic larvae. We incubated X. proxima larvae in petri dishes, with half of the dish covered by black plastic, and placed a light source over the uncovered half of the dish. Initially, all larvae were positively phototactic; however, the percentage of larvae displaying negative phototaxis increased steadily to 50% after 36 hours. As predicted, larval cilia quickly responded to changes in light intensity, straightening as light diminished, and bending as light increased. These results demonstrate that larvae of diverse sponge species coordinate behavioral responses to light using the same mechanism.