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Biological Sciences.

VERTEBRATE ROAD-KILL SURVEY OF THE MOBILE-BAY CAUSEWAY III. David H. Nelson, Department of Biology, University of South Alabama, Mobile, AL 36688. Cynthia Scardamalia-Nelson, Counselor, Providence Hospital, Mobile, AL 36608.

A systematic survey was conducted weekly during 2003 by bicycle (or automobile) on the Mobile Bay Causeway to assess the numbers of vertebrates killed by vehicular traffic. More than 933 organisms were encountered, representing 102 species of vertebrates: 5 amphibians (n=166), 26 reptiles (n=309), 55 birds (n=339), and 16 mammals (n=119). Southern leopard frogs (Rana utricularia) were the most abundant amphibian observed (March to December) representing 73% of all amphibians. "Endangered" Alabama red-bellied turtles (Pseudemys alabamensis, n=99) were the most common reptile, accounting for 32% of all reptiles; most of these (67) were hatchlings. Laughing gulls (Larus atricilla, n=54) and Starlings (Sturnus vulgaris, n=52) were the most frequently encountered birds (representing 16% and 15% of birds, respectively). Raccoons (Procyon lotor, n=42) and Opossums (Didelphis marsupialis, n=36) accounted for 35% and 30% of all mammals, respectively. The most significant finding of this continuing study is the confirmation of 67 hatchlings (March-May, November), 29 adult females (most gravid; May-August), and 3 juveniles (Sept-Oct) of the Alabama red-bellied turtle. Most hatchlings overwintered in the nest to emerge in the early spring. For the last three years, the impact of vehicular mortality on this endangered species has been profound. We also documented the presence of the Florida green water snake (Nerodia floridana) in Baldwin County, Alabama.
 Detection of Total and Pathogenic Vibrio vulnificus using
 Multiplex PCR and DNA Microarrays. Gitika Panicker and Asim K.
 Bej, University of Alabama at Birmingham, AL-35294.

 V. vulnificus, a naturally occurring estuarine microorganism,
 is often found in high numbers in shellfish harvested from the
 Gulf of Mexico during the summer months. It can cause
 gastroenteritis or lead to septicemia and death in susceptible
 individuals when consumed in the form of contaminated raw oysters.
 Results from PCR amplification using oligonucleotide primers for
 regions of 16S rDNA and viuB to identify clinical isolates of V.
 vulnificus showed that 83.3% of environmental isolates had "type A"
 rDNA whereas 61% of the clinical isolates were "type B". On the
 other hand, PCR amplification of viuB followed by gene-probe
 hybridization exhibited positive results for 100% of clinical
 isolates and 22.6% of environmental isolates. Thus, implying that
 viuB is a better target for the identification of clinical strains.
 Next, biotin-labeled multiplexed PCR-amplified viuB and vvh gene
 segments were subjected to microarray hybridization at
 50[degrees]C to detect total and clinical strains. Positive
 hybridizations were detected using Tyramide Signal
 Amplification[TM] with Alexa Fluor[R] 546. Other shellfish-borne
 pathogens, V. parahemolyticus and V. cholerae, were also detected
 using the DNA microarray with their respective gene probes. Results
 exhibit that a combination of multiplex PCR and microarray
 hybridization permits a specific and sensitive system for detection
 of microbial pathogens in shellfish.

URBANIZATION FACTORS THAT AFFECT AQUATIC BIOLOGICAL COMMUNITIES. Janna Owens, Robert Angus, Melinda Lalor, Jaideep Honovar and Ken Marion, University of Alabama at Birmingham, Bham, AL. 35294. Steve McKinney, Storm Water Management Authority, Inc., Bham, Al. 35209.

Urban growth and development in the Jefferson County (Alabama) area are increasing at a steady rate. Factors accompanying urban growth that affect the quality of receiving waters include loss of vegetation, land disturbances, riparian alterations and an increase in impervious surfaces. Impacts related to these changes in water integrity are usually episodic in nature and therefore difficult to profile for water quality evaluations. Our objective was to examine possible correlations between a watershed's urbanization status and the community structures of fish and benthic macroinvertebrates in the upper Cahaba River basin in Birmingham, Alabama. A series of metrics were utilized to evaluate the status of the aquatic communities and to determine possible trends linked to urbanization factors by comparing metrics with water chemistry, habitat conditions, accumulated sediment depths and upstream land usage characteristics. To calculate upstream urbanization characteristics, geographic information systems (GIS) and selected data layers were used to construct a cartographic model. Significant correlations were found between the percentage of impervious surfaces in the upstream watershed and pollution-sensitive macroinvertebrate metrics, such as the EPT and Hilsenhoff Biotic indices. Darters, suckers and selected minnow species correlated negatively with the percentage of impervious surfaces, habitat scores and sedimentation values, respectively. Management strategies for aquatic systems that link water quality monitoring and geographic information with aquatic community data will be able to characterize, detect and remediate the effects of urbanization more precisely.

CONSERVATION GENETICS OF MONTANA SAUGER. Rachael N. Koigi and Neil Billington, Dept. of Biol. Environ. Sciences, Troy State Uni., Troy, AL 36082. William Gardner, Montana Dept. of Fish, Wildlife and Parks, Lewistown, MT 59457.

There is growing concern regarding the decline of sauger (Sander canadensis) populations in Montana. Decline has been greatly attributed to habitat loss, effects of diversion dams, and hybridization with walleye (S. vitreus). No information existed on genetic variation in Montana sauger prior to this study, but 0-15% hybridization has been reported in several previous studies. Protein electrophoresis was used to examine genetic variation in sauger populations and to screen for hybridization between native sauger and introduced walleye. Genetic variation was detected at two (SOD* and PGM-1*) of 25 protein-coding loci screened, excluding the four loci diagnostic between the two species. Although only one heterozygote was found at PGM-1*, significant differences in allele frequencies were found at SOD* in both Missouri River and Yellowstone River sauger populations. This is the first time polymorphism has been reported at the SOD* locus in sauger. Due to the significant genetic variation present in sauger populations in each of the two main river systems, they will need to be managed separately and no stock transfer should be conducted. Hybridization rates compared to those of previous studies (0-20%), being highest in the Missouri river drainage. Hybridization with walleye poses a serious threat to the genetic integrity of the Montana sauger; therefore, brood stock used for supplemental programs should be genetically screened to prevent the unintentional spawning of hybrids.

HYBRIDIZATION AND INTROGRESSION BETWEEN WALLEYE AND SAUGER FROM POOL 13 OF THE MISSISSIPPI RIVER. Jennifer Lynch, and Neil Billington, Dept. Biol. Environ. Sciences, Troy State Univ., Troy, AL 36082, and John Pitlo, Iowa DNR, Bellevue, Iowa 52031.

Iowa Department of Natural Resources (DNR) personnel have observed fish in Pool 13 of the Mississippi River with intermediate morphological characteristics between walleye (Sander vitreus) and sauger (S. canadensis) suggesting they were hybrids. Hybridization and introgression between walleye and sauger has been demonstrated in several studies, which also documented difficulties in determining hybridization rates by morphology compared to protein electrophoresis. Walleye and sauger show fixed allelic differences at four protein coding loci: ALAT* and IDDH* from liver and mMDH-1* and PGM-1* from muscle. Forty-four Sander specimens were collected by electrofishing from Pool 13 on October 22, 2003, 21 (45%) of which were identified by morphology as walleye, 20 (45%) as sauger, and three (7%) as suspected hybrids. Protein electrophoresis of the four diagnostic loci revealed that 17 (39%) were walleye, 16 (36%) were sauger, and 11 (25%) were hybrids. The three fish initially identified as hybrids were confirmed by electrophoresis (two backcrosses to walleye and one [F.sub.1] hybrid). Four fish identified as walleye by morphology contained sauger alleles (two backcrosses to walleye, one backcross to sauger, and one [F.sub.x] hybrid). Four fish identified as sauger by morphology contained walleye alleles (all were backcrosses to sauger by electrophoresis). Overall, 8/44 (18%) fish were misidentified by morphology. The use of protein electrophoresis for identification of Sander specimens is strongly recommended, particularly if hybridization is suspected.

TEMPERATURE DEPENDENT VITELLOGENESIS IN MALE GAMBUSIA AFFINIS. Paul D. Melvin, III, R. Douglas Watson, and Robert Angus Department of Biology, Univ. of Ala. at Birmingham, Birmingham, AL 35294

Many anthropogenic compounds with estrogenic activity have been detected in aquatic environments. Vitellogenin (VTG), an egg yolk protein produced by the liver in oviparous female vertebrates in response to estrogen, is not normally detectable in the serum of males. Various studies have indicated that environmental estrogens can induce vitellogenesis in males of a variety of aquatic vertebrate species, including the western mosquitofish, Gambusia affinis. If VTG presence in male serum is to serve as a reliable estrogen biomarker, it is important to understand other factors that influence its production. Although the regulation of vitellogenesis is poorly understood, previous observations in our lab have suggested that temperature plays an important role. We hypothesize that estrogenic induction of vitellogenin in males will decrease as water temperature decreases.

In this study, male G. affinis were exposed to 17[alpha]-ethynyl estradiol via diet at a concentration of (10 ug/g of food) for a period of one week. This concentration was previously shown to strongly induce VTG production in one week at 24[degrees] C. During exposure, the fish were kept at a constant water temperature of 16[degrees], 20[degrees], or 24[degrees] ([+ or -] 1[degrees]) C. - The animals were then bled by heart puncture. The blood was analyzed by native gel electrophoresis and western blot for the presence of vitellogenin protein in blood serum. Vitellogenin production in male G. affinis was positively associated with environmental temperature. These preliminary results demonstrate that temperature strongly affects the extent to which Gambusia affinis produce VTG in response to estrogen exposure.
 Department of Pharmacal Sciences, Harrison School of
 Pharmacy, Auburn University, Auburn, AL 36849, R. Oswaldo
 Guerrero, School of Pharmacy, University of Puerto Rico, San
 Juan, PR 00936-5067 and Maynard W. Quimby, College of
 Pharmacy, University of Mississippi, deceased.
 Over twenty varieties of Cannabis sativa L. were
 included in our marihuana research program at the University
 of Mississippi. We discovered considerable variation in
 cannabinoid content of marihuana samples. Factors affecting
 cannabinoid content included heredity, plant parts in the
 sample, age of plants when harvested.and environmental
 factors. [DELTA]9-tetrahydrocannabinol (THC) content was
 highly variable among these varieties. Those with higher THC
 values were identified as "drug types." They often were
 associated with drug use. Those with low THC content were
 identified as "fiber types." They often were associated with
 plants grown for fiber (hemp).
 Although most of our studies focused on drug types,
 fiber types remained of interest as potential agricultural
 crops. There was a need to determine how much the cannabinoid
 content varied during the growing season in a fiber type. A
 Turkish fiber variety was selected. The results of this study
 will be shared. Comparisons of drug and fiber types also will
 be made in this illustrated presentation.

NESTING ACTIVITY IN THE ALABAMA RED-BELLIED TURTLE (PSEUDEMYS ALABAMENSIS). Gabriel J. Langford, David H. Nelson, and Joel A. Borden, Dept. of Biology, Univ. of S. Ala., Mobile, AL 36608.

A study of nesting activity in the Alabama red-bellied turtle was conducted in the vicinity of the Mobile Bay causeway (Baldwin Co.) during the summer of 2003. Females were observed as they deposited eggs in the field. Nests were labeled and protected by predator-exclusion covers to conserve eggs until they could successfully complete development. Nests were laid along terrestrial elevations from June 15 to August 5, 2003. Clutch sizes for 20 nests ranged from 7 to 19 eggs (mean = 12.0). Developmental periods (from oviposition to emergence) for nests ranged from 67 to 123 days (mean = 99.2). The numbers of hatchling emerging from each of the nests ranged from 2 to 14 (mean =8.4). The numbers of undeveloped eggs ranged from 0 to 17 eggs per nest (mean =3.5). Virtually all hatchlings manifested maxillary cusps, eyebars, and reddish plastrons (with vermiculations). Measurements of 177 hatchlings (from 21 clutches) disclosed a mean carapace (straight line) length of 3.7cm, a mean carapace width of 3.6cm, a mean plastron length of 3.4cm, a mean tail length of 0.97 cm, and a mean wet body weight of 11.9 grams. Predation of uncovered nests by fish crows, raccoons, and ants was very high. At one site on the Blakely River, the ratio of depredated to covered nests was 13:1. A total of 159 protected hatchlings were released into the environment.

USE OF ANAEROBIC BACTERIA AS PROBIOTICS IN AQUACULTURE. Mark Meade and Benjie Blair. Jacksonville State University, Dept. of Biology, Jacksonville AL 36265.

Probiotics have rarely been used in aquaculture. In this study, we examined the effects of an anaerobic bacterium as a potential probiotic in the culture of tilapia fry. Nile tilapia fry (0.20-0.25 g) we held in three 260l fiberglass tanks with associated re-circulating biofilters (n=200 fish/tank). All fish were fed a high protein commercial fish ration that contained a minimum of 50% crude protein (Aquamax[R] Starter Fingerling 300, PMI Nutrition International, Inc.). Feed proffered to two of the tanks was initially supplemented with anaerobic bacteria. Specifically, 25ml of E. cellulosolvens cells in exponential growth phase were collected by centrifugation and mixed with the feed. On subsequent feedings, all fish were fed solely the high protein fish ration not containing bacteria. Survival of tilapia in each tank was >95%. Final mean wet weights, however, were significantly different between the two treated tanks compared to the control tank. Specifically, fish supplemented with the bacteria had final mean wet weights of 0.99g [+ or -] 0.6 (s.e.) and 0.83g [+ or -] .05 (s.e.), whereas the control fish had final mean wet weights of 0.62g [+ or -] 0.3 (s.e.). An analysis of size distribution frequencies demonstrated that between 40 and 50% of bacteria supplemented fish were 1 g or larger in size, whereas < 20% of the control fish had attained a size of 1g. These data suggest that anaerobic bacteria may be a beneficial probiotic in the culture of tilapia and may serve to accelerate growth rates.

FLORIDA MANATEE FINGER BONES UNDER THE FINGERNAILS. Gerald T. Regan, Chief Scientist, Marterra Foundation Inc. 4000 Dauphin Street Mobile, AL 36608.

The nails of specimen SHCM350 had the second, third, and fourth finger bones underlying them. The finger bones are counted as though the homolog of the thumb is the first. This observation is reported here because the most recent reviews of the subject of the nails of manatees appear to be silent on the finger bone issue. There have been reports of Florida manatees with four nails, and this report can make it easier to describe which of the remaining finger bones underlies a fourth nail when present. The opportunity to make the observation occurred when a male Florida manatee, Trichechus manatus latirostris, 260 cm long, stranded dead on the south side of Dog River in Mobile County on February 25, 2002. The cause of death was probably cold stress. After the necropsy I froze the flippers, in which homologs of nearly all of the bones of a mammalian forearm are located. Each had three nails on its dorsal surface. On June 14, I took the frozen right flipper to the office of a chiropractor to have X-ray photographs made after positioning X-ray opaque hex nuts on each nail. Once the X-ray film was developed, there was no doubt about the conclusions. I acknowledge the help of Cheryl A. King, the funding of the transportation of the manatee by the USFWS, and the donated expert X-ray service of Jeff Townley and his staff.

EVALUATION OF DEVELOPMENTAL TOXICITY OF INTERACTION BETWEEN CAFFEINE AND PSEUDOEPHEDRINE USING FROG EMBRYO TERATOGENESIS ASSAY-XENOPUS (FETAX). Bernice L. Moser, Angellete Daegle, and James R. Rayburn, Dept. of Biology, Jacksonville State University, Jacksonville, AL, 36265-1602.

Pseudoephedrine and caffeine are found in many over-the-counter drugs including decongestants and weight loss chemicals. Both are proven central nervous system stimulants, and there has been much discussion about potential interactions of these chemicals on human health. Frog Embryo Teratogenesis Assay-Xenopus (FETAX) was used to determine the developmental toxicity of pseudoephedrine and caffeine mixtures and to determine if synergism or antagonism occurs between the two. FETAX is a 96-hour developmental toxicity assay that screens for direct acting teratogens. Both have both been evaluated for developmental toxicity in FETAX but not as mixtures. Potentiating effects of caffeine on the cardiovascular teratogenicity of ephedrine in chick embryos has been shown. Due to the similarities between pseudoephedrine and ephedrine we wanted to determine if synergism occurred between pseudoephedrine and caffeine. The 96-hr LC50, 96-hr EC50, MCIG, and TI were determined for pseudoephedrine and caffeine in various mixtures. We tested five binary mixtures of the two chemicals. The mixtures were based on the toxic units of each chemical; where one toxic unit was equal to the 96-hr LC50. The toxic unit mixtures tested was pseudoephedrine to caffeine at toxic unit ratios of 0:1, 1:0, 3:1, 1:1, and 1:3. The 0:1 and 1:0 test the individual chemicals by themselves. Toxic units were plotted on an isobole graph to determine if synergism, concentration response or antagonism occurred

AQUATIC HERPETOLOGICAL INVENTORY OF THE UPPER MOBILE-TENSAW DELTA Joel A. Borden, David H. Nelson, Gabriel J. Langford, Dept. of Biology, Univ. of South Ala., Mobile, AL 36688.

A herpetological field survey was conducted during the summer of 2003 on the Upper Delta Wildlife Management Area in Mobile and Baldwin Counties, Alabama. Sampling techniques included minnow traps, dipnets, hand captures, visual surveys, cryptozoan (cover) boards, anuran vocalization, and PVC tubing (for tree frogs). A total of 14 species of amphibians were encountered (N=1441): 3 salamanders (all aquatic) and 11 anurans. Most frequently encountered were bronze frogs, bullfrogs, bird-voiced tree frogs, and gray tree frogs. Minnow traps were the most effective sampling technique for amphibians. No terrestrial salamanders were ever recorded (even under cover boards). A total of 24 species of reptiles were encountered (N=372): 7 turtles, 1 crocodilian, 4 lizards, and 12 snakes. Most frequently recorded were black-knobbed sawback turtles, American alligators, ground skinks, five-lined skinks, ribbon snakes, cottonmouths, and banded water snakes. Visual observations proved to be the most productive technique for reptiles. Five glossy crawfish snakes and 3 mud snakes were captured by hand or in minnow traps.

THE EFFECT OF URBANIZATION ON THE DIVERSITY OF FISH ASSEMBLAGES AND THE BIOLOGICAL INTEGRITY OF IMPACTED STREAMS IN THE SOUTHEASTERN PLAINS ECOREGION, ALABAMA. Bonnie Hamiter, Kristy Pisani, Christa Collins, Megan Pilarczyk, and Jonathon Miller, Department of Biological and Environmental Sciences, Troy State University, Troy, AL 36081.

One of the leading causes of stream impairment in the United States is urbanization. By assessing biological communities, previous studies have shown the negative impact of urbanization on biological integrity. Urbanization affects the presence of intolerant and tolerant species, species diversity, and the occurrence of deformities, erosions, lesions, and tumors (DELTs) Eighteen urban-impacted and three least-impacted streams in the Choctawhatchee and Pea Rivers watershed were sampled for fish assemblages using a backpack electroshocker. Of the 3,654 fish collected from the 21 sample sites, 38 species were identified. Shannon-Wiener diversity index results did not indicate lower species diversity at urban-impacted sites than least-impacted sites. Data analysis also included application of an Index of Biotic Integrity (IBI) previously established for the Choctawhatchee and Pea Rivers watershed. The IBI did not show reduced scores at urban sites. However, certain metrics within the IBI revealed an impact on the fish assemblages in urban sites. Examination of DELT anomalies suggested a greater abundance of lesions and erosions at urban than least impacted sites. Results suggest that the IBI established for the study area may only adequately assess the stream biological integrity for the stream conditions under which it was established.
 J. Doorenbos, Department of Pharmacal Sciences, Harrison
 School of Pharmacy, Auburn University, Auburn, AL 36849,
 James Flor, Merck Sharp and Dohme Laboratories, Rahway,
 NJ, deceased, Larry Robertson, College of Pharmacy, Ohio
 State University, Columbus, OH 43210.
 Dehydroabietic acid, readily available from natural
 sources. is a potential starting material for the
 synthesis of Carylsteroids.and other substances of
 potential medicinal value. Its utility would be enhanced
 by the presence of additional functional groups. This
 research was undertaken to find fungi to introduce
 hydroxyl groups.
 We wish to report the discovery that submerged cultures
 of Aspergillus flavipes and Whetzelinia sclerotiorium may
 be used to prepare 7[beta]-hydroxydehydroabietic acid,
 7[beta],15-dihydroxydehydroabietic acid and
 3[beta],7[beta],15-trihydroxydehydroabietic acid and
 derivatives. These cultures were selected after screening
 twenty species of fungi in the first phase of this study.
 Chemical structures of the metabolites were established
 by physical and chemical methods including n.m.r., m.s.,
 i.r. and elemental analyses.

FISHING FOR MERCURY AT THE LOCAL GROCERY. Dustin W. Morin, Karla E. Hinds, and Alfred C. Nichols, Dept.of Physical and Earth Sciences, Jacksonville State University, Jacksonville AL 36265.

Fish consumption is considered the single largest source of mercury (Hg) exposure to humans. Methyl mercury, the form of the metal most commonly found in fish, is both a human teratogen and central nervous system toxicant. We have tested samples of canned, frozen, and fresh fish purchased from local grocery stores for Hg concentrations. Samples were analyzed according to the USEPA Manual Cold Vapor Technique. Except for one sample of fresh catfish fillet, Hg was detected in all fish samples tested. Mercury levels in canned fish ranged from 0.01 to 0.51 ug/g (dry wt.). Highest concentrations of the metal were found in frozen swordfish steak (0.97 ug/g dry wt.) and frozen shark steak (0.76 ug/g dry wt.). While below the FDA limit, such levels may be of concern to young children and women in their reproductive years. Also included in the survey was a sample of canned chicken (no Hg detected) and a fillet from a channel catfish caught in a local farm pond (Hg level of 0.09 ug/g dry wt.).

A REVIEW OF THE CHEMICAL ECOLOGY OF ANTARCTIC MARINE SPONGES. James B. McClintock and Charles D. Amsler, Dept. of Biology, Univ. of Ala. at Birmingham, AL 35294. Bill J. Baker, Dept of Chemistry, Univ. of South Fla., Tampa, FL 33620. Robert Van Soest, Univ. of the Netherlands.

The continental shelf waters of Antarctica are dominated by rich sponge communities. High in terms of both species diversity (417 species identified to date) and in biomass, these communities contribute considerable structural heterogeneity and surface area for epibiotic organisms. They also provide a significant food resource to predators, especially sea stars, whose selective feeding has important effects on community structure. Brief seasonal periods of mesoplankton availability are paradoxical considering the filter feeding habits of sponges. Food resources may be supplemented by uptake of DOM, and capture of smaller nanoplankton and picoplankton. In contrast to earlier theories that due to the lack of fish predation high latitude sponges should be depauperate in defensive metabolites, chemical defenses in Antarctic sponges are not uncommon. A variety of secondary metabolites have deterrent effects against spongivorous sea stars, and may be optimally sequestered in the outermost layers of sponges to deter their extra-oral feeding. Sponge metabolites have also been found that short circuit molting in sponge-feeding amphipods and prevent fouling by benthic diatoms. Coloration is a select group of Antarctic sponges may be the result of relict pigments originally selected for temperate or tropical conditions as aposomatics or UV screens that have been conserved because they themselves have defensive properties. Supported by NSF OPP-9814539 and OPP-0125181 to J.B.M and C.D.A. and OPP-990176 and OPP-0125152 to B.J.B.
 CREEK. Jeremy White, Reed Hogan, L. J. Davenport, Department of
 Biology, Samford University, Birmingham, AL 35229.

 Invasive species have had a dramatic effect on the ecosystem
 surrounding Shades Creek. This 55-mile long waterway starts in
 Irondale, Alabama and ends in northern Bibb County as it flows
 into the Cahaba River. Four sites were chosen for this study, each
 varying in the level of human development present. Each site was
 studied using transect sampling with a 25-meter transect on both
 the north and south bank. All species were catalogued that lay
 within a 12-inch distance to either side at each meter mark. A
 total of 64 species were catalogued: 50 native species and 14
 non-native species. Of the non-natives, three were deemed to be
 highly invasive--Kudzu, Chinese Privet, and Mimosa. This study
 found a direct correlation between the increase in human
 development and the increase in the presence of invasive species.
 (The types of human development included commercial and
 residential buildings, roadways, utility lines, and recreational
 structures such as trails.) As human development increases, there
 is an increase in the percentage of invasive species, a decrease
 in biodiversity, and a decrease in the presence of the native
 flora that is indigenous to that particular area.

ANALYSIS OF PCB CONTAMINATION IN LARGE MOUTH BASS AND CHANNEL CATS OF LOGAN MARTIN LAKE. Shawn Bailey and James R. Rayburn, Jacksonville State University, Jacksonville, AL 36265-1602, USA

This project will screen the fresh water species of Logan Martin Lake for possible PCB contamination with a qualitative analysis. The target species will consist of one bottom feeder, the channel cat (Ictalurus punctatus), and one predator, the large mouth bass (Micropterus salmoides). Composite samples will be taken of both species for data correlation between other testing groups and to increase the integrity of the results. These meet with the parameters set forth by the EPA Fish Contaminant Workgroup and the Alabama Department of Environmental Management (ADEM) and are representative of habitual consumptive habits for the local recreational anglers. If initial tests are positive, the samples will be retested with a quantitative analysis and weighed against the EPA's screening values and the FDA's action levels to ascertain the degree of risk associated with fish consumption in the areas tested. This is the first step in assessing a contamination risk to the recreational consumer, one that would support the fish advisories and tests already issued by the Alabama Department of Public Health (ADPH), EPA, and ADEM. This is also the first step towards a bioremediation project.
 Alyssa Geis. Thane Wibbels, Dept. of Biology, Univ. of Ala.
 at Birmingham, Birmingham, AL 35294. Dawn Fletcher, William Gates,
 and Randy Swilling, Bon Secour National Wildlife Refuge, Gulf
 Shores, AL 36542.

 The loggerhead sea turtle (Caretta caretta) is a protected
 species inhabiting the Gulf and Atlantic coastal waters of the
 United States. Previous studies have identified several distinct
 genetic groups of loggerheads in these waters. The current project
 evaluated the origins of juvenile loggerhead sea turtles inhabiting
 the coastal waters of Alabama and hatchling loggerheads from nests
 monitored at the Bon Secour National Wildlife Refuge (BSNWR). To
 determine the genetic stock of these turtles, mitochondrial DNA
 control region sequences were sequenced and analyzed from juvenile
 and hatchling turtles. Blood samples were obtained from juvenile
 loggerheads that were captured by tangle net in the Alabama bay
 systems. Tissue samples were obtained from hatchlings that were
 found dead in nests at BSNWR after all the live hatchlings had
 emerged. Preliminary analysis of the data suggests that these
 turtles (both juvenile and hatchling) represent haplotypes common
 to the current management units referred to as the South Florida
 nesting subpopulation (occurring from northeast Florida to Sarasota
 on the west coast of Florida) and the Florida Panhandle nesting
 subpopulation (occurring at Eglin Air Force Base and the beaches
 near Panama City, Florida). The results of this study have
 significant management implications for loggerhead sea turtles in
 the northern Gulf of Mexico.

AN UPDATE ON FUNGAL INFECTIONS ASSOCIATED WITH AMBROSIA BEETLE INFESTATIONS. Roland R. Dute, Brian L. Prather and Matthew J. Valente Dept. of Biological Sciences. Kathy S. McLean, Dept. of Entomology and Plant Pathology, Auburn University, Auburn AL 36849

The Asian ambrosia beetle (Xylosandrus crassiusculus) carries the fungus Ambrosiella xylebori with which the insect infects the wood of its host. The fungus, as it grows, serves as a food source for the beetles. In the spring of 2003, we were unable to isolate A. xylebori from either infected wood showing wilt or from beetles trapped in flight. There is controversy as to whether the activities of A. xylebori or a secondary fungus, also introduced by the beetle, are responsible for foliage wilt of trees. We decided to take a common fungal isolate from infested trees, Fusarium oxysporum, and attempt re-infection of a host (Cercis canadensis, redbud). Eight, saplings were used: two untreated controls, two receiving drill holes without inoculum, and four receiving drill holes with inoculum. After seven weeks the saplings were felled and 2 cm thick wood disks were cut at 2 cm above each drill hole. One-half of each disk was processed for fungal growth and culturing, and the other half was preserved and later sectioned with a sliding microtome. Although the saplings were asymptomatic at the time of collection, culturing showed the presence of F. oxysporum only in inoculated wood disks. This observation was confirmed using light microscopy on sectioned material. The presence of hyphae was distinct from the induction of brown discoloration and plugging of the water-conducting cells induced by the act of wounding the trees.

INFLORESCENCE ANATOMY OF COMMELINA ERECTA L. Roland R. Dute, Department of Biological Sciences, Auburn University, Auburn, AL 36849. Floyd Woods, Department of Horticulture, Auburn University.

It is thought that various floral parts represent modified leaves. In Commelina erecta each flower contains both petals and sepals of two different sizes, and each inflorescence (flower cluster) is enclosed by a leaflike spathe. The anatomy of these laminar organs was compared with that of the vegetative leaves. Venation is closed in leaves, spathes, and small sepal; partly open in the large sepal; and open in the large petals. Vein terminations in large petals and (to a lesser extent) in large sepals consist of bundle sheath cells that extend beyond xylem and phloem. In petals, these vein prolongations exist as branched, digit-like structures. Callose staining indicates numerous cytoplasmic connections between bundle sheath cell prolongations and neighboring non-vascular cells. It is thought that the prolongations represent an auxiliary transport system shunting food and water to very thin portions of the petal. Elongate, multicellular raphide sacs form parallel rows in all organs investigated. Stomata, associated with gas exchange, are also located on all organs investigated. On leaves, spathes, and sepals, stomata consist not only of guard cells, but also of subsidiary cells. Petals however, have stomata with only guard cells. Elongate sacs containing granular or amorphous ergastic material exist in petals and sepals. Histochemical staining shows this material to be tannin. It is hypothesized that these sacs serve as deterrents to herbivory. In summary, the different laminar organs show different combinations of features.

Effect of Cyclic AMP on Protein Metabolism in Crustacean (Callinectes sapidus) Y-Organs. Deug Woo Han and R. Douglas Watson, University of Alabama at Birmingham.

Paired Y-organs secrete ecdysteroid hormones that control cycles of growth and molting in crustaceans. Y-Organs are regulated, at least in part, by molt-inhibiting hormone (MIH), a peptide produced and released by the X-organ/sinus gland complex of the eyestalks. We are investigating cellular signaling pathways involved in regulation of Y-organ function. In the present studies, Y-organs were incubated in vitro in the presence of [.sup.35] S-methionine ([.sup.35] S-met) and experimental agents that influence the cyclic AMP (cAMP) signaling pathway. In 4-hr incubations, 8-Br-cAMP (a cAMP analog) suppressed incorporation of [.sup.35] S-met into Y-organ proteins; the effect was concentration-dependant. Addition of cholera toxin (a Gs protein activator), IBMX (a phosphodiesterase inhibitor), or forskolin (an adenylate cyclase activator) likewise suppressed incorporation of [.sup.35] S-met into Y-organ proteins. The effect of cholera toxin was antagonized by KT5720 (a protein kinase A inhibitor). Incorporation of [.sup.35] S-met into Y-organ proteins was also suppressed by an extract of sinus glands. The combined results suggest that cAMP plays an important role in regulation of Y-organ function. We are currently investigating the link of protein synthesis to ecdysteroid production, and the possibility of cross-talk between cAMP and other cellular signaling pathways in Y-organs. Supported by the National Science Foundation (IBN-0213047).

NOTES ON THE BIOLOGY OF THE FRESHWATER JELLYSFISH (CRESPEDICUSTA SOWERBII) IN NORTHEASTERN ALABAMA. George R. Cline, James R. Rayburn, Mijitaba Hamissou, and Frank A. Romano. Biology Dept. Jacksonville State University, 700 Pelham Road North, Jacksonville, AL 36265-1602.

The freshwater jellyfish (Craspedacusta sowerbii) a hydrozoan with a complex lifecycle and a cosmopolitan distribution. Little is known about the biology and ecology of this organism, as it's distribution is patchy, and it's occurrence is strongly seasonal. In Alabama, C. sowerbii is documented from 15 sites in 13 counties. The study site is a flooded quarry in northeastern Alabama. A sharp thermocline is found between 25-40 ft deep, where jellyfish tend to congregate around midday. Conductivity, turbidity, pH, and dissolved oxygen were collected at the surface, 15, and 30 ft using a water quality probe. Jellyfish were collected with plastic storage bags by scuba divers and returned to the lab to observe behaviors, to collect life history data, and for molecular and genetic analyses. Jellyfish first appeared in late June or early July. Population densities peaked in August though October. Numbers dropped dramatically in the late October and early November samples before the dive site closed in late-November. Gonadal tissue was collected from specimens collected from Aug. - Sept. All specimens had active sperm in at least one gonad. These specimens were then prepared for protein, genomic, and mtDNA analyses using RFLP. Some specimens were maintained in 10 gallon aquaria and fed brine shrimp. Jellyfish alternated bouts of swimming vertically with variable length periods of resting on the bottom of the tank. Videotape demonstrating feeding activity is presented.

CARAPACE SHAPE AMONG MULTIPLE TURTLE SPECIES. David A. Delecki Jr., Anissa Delecki, and George Cline, Jacksonville State University, Jacksonville, Al, 36265.

Turtles are one of the most common groups of reptiles in the world. Modern turtles are found in aquatic, marine, or terrestrial habitats and are distributed worldwide in tropical and temperate environments. Turtles are also one of the most unusual groups in that they possess a hard shell. The bony shell of turtles is composed of the carapace and the plastron joined together by a bridge. The carapace is the top part of the shell and is covered by a layer of scales called scutes. The plastron is the bottom of the shell that protects the ventral side of the turtle. The carapace of turtles comes in many different shapes and sizes. Most studies that have examined carapace size in turtles have involved sexual dimorphisms. In a majority of studies a single character (i.e. carapace length) has been used to focus on broad comparisons among species. In studies involving more than one carapace measurement usually only one turtle species is used. In this study calipers were used to measure the carapace length, anterior carapace width, posterior carapace width, anterior carapace height, and posterior carapace height. These measurements were then analyzed to test for patterns related to the ecology and phylogeny of turtles.

DOES ENVIRONMENTAL STRESS INCREASE FLUCTUATING ASYMMETRY IN THE EASTERN MOSQUITOSIH? Robert A. Angus and Eleanor J. Estes, Dept. of Biology, Univ. of Ala. at B'ham., Birmingham, AL 35294.

Fluctuating asymmetry (FA), defined as random differences between the left and right sides of a normally bilaterally symmetrical organism, has been used in numerous studies as an indicator of developmental instability, one possible cause of which is stress. This study focuses on two populations of eastern mosquitofish (Gambusia holbrooki) from small Florida coastal streams. Fish inhabiting the Fenholloway River live in a chemically polluted environment as a result of effluent from a paper mill. These fish presumably experience much more stress during development that those in unpolluted streams. The control fish were collected from Spring Creek, a nearby nonpolluted stream that does not receive effluent from a paper mill. In order to compare levels of FA, 9 morphometric (measured) and 5 meristic (counted) traits were measured on the left and right sides of 26 mature females collected from each site. The measurements were taken by one person, in order to minimize differences in personal bias and were taken twice in order to permit estimation of measurement error. To analyze the data, statistical methods developed for studies of FA by Palmer and Strobek were used. The methods include tests for aberrant individuals, outliers, measurement error, asymmetry and FA. The analyses for FA assess the differences between individuals, the two populations of fish and also take into account how the varying size of the fish and the trait affect FA. This study tests the hypothesis that stress caused by environmental pollution produces increased FA in mosquitofish. If correct, then FA could serve as a useful bioindicator of populations experiencing pollution-induced stress.

THE USE OF DUAL-ENERGY X-RAY ABSORPTIOMETRY TO DETERMINE BODY COMPOSITION IN FISH. Randy J. Watts and Stephen A. Watts, Dept of Biology, Univ. of AL at Birmingham, Birmingham, AL 35294. Hugh S. Hammer, Aquatic Sciences Dept, GSCC, Gadsden, AL 35902. Maria S. Johnson and Tim R. Nagy, Nutrition Science, Univ. of AL at Birmingham, Birmingham, AL 35294.

Body composition analysis provides information on energy allocation and utilization from many animal models. The use of non-invasive techniques for determining body composition, including bioelectrical impedance, electrical conductivity, body condition indices and isotopic dilution have shown limited success for determining fat, lean and ash content in many animal models, but have been validated for human and some smaller mammals. Fat composition of fish can be a useful predictor of body condition, but evaluation has previously required destructive sampling. Dual-energy X-ray absorptiometry (DXA) was originally designed to detect osteoporosis in humans. DXA can also be used to determine other components of body composition such as fat mass and lean tissue mass. Recent studies indicated that DXA can be used to accurately determine fat and lean composition in tilapia. Trials are underway to evaluate DXA for analysis of catfish. The use of DXA would allow longitudinal studies with individual subjects under a variety of experimental conditions. DXA could provide a valuable tool for finfish analysis and result in optimal management strategies for aquacultured species.

INHIBITION OF hTERT IN HUMAN BREAST CANCER CELLS USING RNA INTERFERENCE. Amanda P. Cunningham, Liang Liu, Rebecca Wylie, Sharla Phipps, Lucy Andrews, and Trygve Tollefsbol, Dept. of Biology, UAB, Birmingham, AL 35294.

RNA interference is a relatively recent innovation that possesses potential as a therapeutic agent for a variety of diseases either alone or in conjunction with existing therapies. We have chosen to use RNA interference as a means to knock down hTERT expression in the estrogen receptor negative human breast cancer cell line MDA-MB-157. hTERT is the catalytic component of the enzyme telomerase which is expressed in >90% of human cancers, but with few exceptions is not expressed in normal somatic cells. We believe this selective quality makes hTERT an attractive therapeutic target and RNA interference a promising means of achieving hTERT knock down in human cancer cells. We have shown hTERT mRNA to be effectively knocked down in MDA-MB-157 cells by all three of our preliminary retroviral constructs for shRNA delivery by RT-PCR analysis after 8 weeks of treatment.

AROMATASE mRNA LEVELS IN THE BRAIN OF TRACHEMYS SCRIPTA. Keela L. Dodd, Chris Murdock, and Thane Wibbels, Department of Biology, Birmingham, AL 35294-1170.

Many reptiles exhibit temperature-dependent sex determination (TSD), in which the incubation temperature of the egg determines the sex of the developing embryo. The physiology of TSD is not well understood; however, estrogen has been hypothesized to play a key role. One hypothesis suggests that female incubation temperatures result in the increased production of mRNA for the enzyme cytochrome P450 aromatase that catalyzes the conversion of androgens to estrogens. Further, it has been suggested that the brain may be the source of aromatase activity prior to or during sex determination. In the current study, a quantitative-competitive RT-PCR was validated for quantifying the aromatase mRNA levels in the brain tissue of Trachemys scripta, a turtle with TSD. Eggs were incubated at male- and female-producing temperatures, and tissues were harvested at developmental stages before, during, and after the thermosensitive period of TSD. A quantitative-competitive RT-PCR was preformed using a RNA competitor as an internal standard in each reaction tube. The competitor molecule was a fragment of the endogenous aromatase mRNA that contained a deletion, allowing it to be differentiated from endogenous aromatase using gel electrophoresis. The intensity of the endogenous and competitor bands were measured digitally and then used to estimate the amount of endogenous aromatase in each sample. All examined tissues had low or non-detectable levels of aromatase. Further, no significant differences were found between embryos at male versus female temperatures. The results do not support the hypothesis that brain aromatase expression plays a pivotal role in TSD.


A land-developing company is in the process of building a 600-acre development in the Shannon Valley. The construction appears to be dumping large amounts of sediment into Ross Creek, which in turn flows into Shades Creek. When observing the stream, there is no evidence of benthic macroinvertebrates or bottom-laying fish, and the natural environment around the stream has been disturbed by the removal of riparian vegetation. The sediment from the construction is contributing to the poor stream life of Ross Creek. To prove this hypothesis, one-liter water samples were taken from five sites along Ross Creek. Samples were taken on two days during rainfall and one day when it had not rained for three days. The samples were tested for total suspended solids and total dissolved solids. The results showed that the levels of suspended and dissolved solids from every sample site on Ross Creek increased after rainfall as opposed to dry periods. In some instances, the levels increased as much as 500%. Every sample taken from Ross Creek had higher levels of solids than same-day samples taken from Shades Creek upstream from the entry point of Ross Creek. The construction is disrupting the aquatic environment of Ross Creek. The company developing the land is responsible for making sure that the water is not degraded, and it needs to take proper action to stop runoff into the waterway. Down-Regulation of Telomerase in Human Promyelocytic Leukemia Cells and Estrogen Receptor Negative Breast Cancer Cells by Epigallocatechin-3-Gallate (EGCG). Joel B. Berletch, Mitchell S. Pate, Lucy G. Andrews, Trygve O. Tollefsbol. University of Alabama at Birmingham.

Aging can be characterized as a series of time-dependent physiological and anatomical changes that reduce the capacity to adjust to various stresses presented in life. One such age related stress is cancer. Studies have shown that 85-95% of cancer cells over-express a ribonucleoprotein called telomerase. Telomerase expression is necessary to immortalize cancer cells by maintaining the ends of chromosomes therefore preventing attrition. The polyphenolic compound epigallocatechin-3-gallate (EGCG) has been shown to decrease telomerase activity in cancer cells. Many mechanisms have been proposed but none have been well established. It has been shown that EGCG has the capacity to down-regulate telomerase in an estrogen receptor positive breast cancer cell line, MCF-7. However down regulation of telomerase in promyelocytic leukemia cell line HL-60, due to the effects of EGCG, has not been well documented. By performing the telomeric repeat amplification protocol (TRAP assay), this study demonstrated that EGCG has a down-regulating effect in Leukimia cells. Further studies involving EGCG and current cancer treatments may be helpful in elucidating the mechanism utilized by EGCG.

A PROPOSED INDEX OF BIOLOGICAL INTEGRITY FOR AQUATIC MACROINVERTEBRATES. L. J. Davenport and W. Mike Howell, Department of Biology, Samford University, Birmingham, AL 35229.

While aquatic macroinvertebrates have long been used to assess the health of streams, different investigators choose different metrics to determine that health. Current "popular" metrics include the total number of taxa and the number or percentage of organisms in specific groups, such as EPT (Ephemeroptera, Plecoptera and Trichoptera), chironomids, and the most dominant taxon. In contrast, Hilsenhoff's (1987) Biotic Index uses tolerance values established for all taxa, and an average tolerance value is computed. In a manner similar to Karr's (1981) Index of Biological Integrity for fishes, we attempted to combine the above metrics (plus others) into a single index for aquatic macroinvertebrates. Sample data utilized is from our 2001-2003 studies at fifteen sites on the upper Cahaba River.

Detection Vibrio parahaemolyticus O3:K6 using real-time PCR with Taqman fluorescent probes in Gulf of Mexico water and shellfish. Amy V. Rizvi and A. K. Bej, University of Alabama at Birmingham, AL-35294.

Vibrio parahaemolyticus is a Gram-negative bacterium and a natural inhabitant of warm costal waters. It is commonly found in shellfish and other crustaceans, and is responsible for causing gastroenteritis when consumed in raw or poorly cooked seafood. Conventional microbiological methods of detection of this pathogen are relatively time consuming. Therefore the need for rapid, more accurate detection of pathogens in seafood has lead to the development of a multiplex real-time PCR assay. This assay was developed by optimizing the use of Taqman probe-based multiplexed PCR parameters and testing tl and ORF8 gene-specific oligonucleotide primers and probes in an effort to establish a rapid, specific and sensitive real-time method of detection of total Vibrio parahaemolyticus and the pandemic serovar O3:K6 in oyster tissue homogenate and Gulf of Mexico water. The specificity of these primers was tested for amplification of a 450 bp tl and a 369 bp ORF8 gene fragment representing all V. parahaemolyticus and post-1996 clinical isolates of pandemic serovar O3:K6, respectively. The sensitivity of detection was 10 pg of purified genomic DNA and 1[0.sup.3] cfu in 1 ml in pure culture. Enrichment of the seeded samples of both oyster tissue homogenate and gulf water for 5 h or 8 h resulted in a sensitivity of detection level of 1 cfu in 1 g of seeded enriched oyster tissue homogenate or 1 ml gulf water. Rapid, reliable and sensitive detection of V. parahaemolyticus O3:K6 would help the shellfish industry and ISSC in taking appropriate measures for routine monitoring for this pathogen, thereby preventing disease outbreaks and consequently protecting consumer health.

INCUBATION TEMPERATURES IN GREEN TURTLE NESTS AT FRENCH FRIGATE SHOALS, HAWAII. Jennifer Estes, Thane Wibbels, Dept. of Biology, Univ. of Ala. at Birmingham, Birmingham, AL 35294. George Balazs, National Marine Fisheries Service, Honolulu, Hawaii.

The green sea turtle (Chelonia mydas) possesses temperature-dependent sex determination (TSD) in which the incubation temperature of the egg determines the sex of the hatchling. This form of sex determination is of evolutionary significance because it has the potential of producing a wide variety of sex ratios. Hatchling sex ratios resulting from TSD are also of conservational and ecological interest, since they can affect the recovery of endangered populations. French Frigate Shoals (FFS) is an atoll located approximately 800 km northwest of Oahu in the Hawaiian Archipelago. Over 90% of Hawaiian green turtle nesting occurs on FFS. The purpose of this study was to evaluate nest temperatures in an effort to estimate sex ratios. Sand and nest temperatures were monitored on FFS during the 1998-2002 nesting seasons. The average incubation temperatures during the middle third of incubation were used to predict sex ratios. The pivotal temperature of sex determination for Hawaiian green turtles is unknown. If the pivotal for Hawaiian green turtles is similar to that of green turtles in Suriname and Costa Rica, where the pivotal temperatures have been estimated, then the majority of temperatures recorded in this study would be below the pivotal. This would suggest an overall male-bias. Alternatively, the Hawaiian green turtles may have evolved a lower pivotal temperature, which could result in unbiased or even female-biased sex ratios at these temperatures.

ZEBRAFISH GILLS: LOCALIZATION OF ALKALINE AND ACID PHOSPHATASES. Chen Jiang and Glenn M. Cohen, Dept. Bio. & Environ. Sci., Troy State Univ., Troy, AL 36082. Eric G. Spokas, Dept. of Chemistry, Rowan University, Glassboro, NJ 08028.

Fish gills continuously perform respiratory, excretory, and ionoregulatory functions. Although enzymatic activities of fish gills have been extensively studied, particularly N[a.sup.+]/[K.sup.+]-ATPases, scant attention has been paid to alkaline and acid phosphatases, two useful marker enzymes for structural and functional studies. Our objectives in the present study were to map the localization of alkaline and acid phosphatases in the zebrafish gills. We used a napthol AS-MX phosphate-stabilized diazonium salt for demonstrating alkaline phosphatase activity (fast blue BB) and acid phosphatase activity (fast red violet LB). We embedded in Spurr epoxy and cut 1-2 [micro] sections. Purified methyl green served as a cytoplasmic and nuclear counterstain. We found sharply contrasting staining patterns for alkaline and acid phosphatases in primary and secondary lamellae. For example, alkaline phosphatase intensely stained the secondary lamellae; all cell types stained. Generalized alkaline phosphatase staining stopped abruptly at the bases of the secondary lamellae where they connect to the primary lamellae. By contrast, acid phosphatase strongly stained cells in both the primary and secondary lamellae as evidenced by the presence of reaction produce sprinkled cytoplasmically. Acid phosphatase displayed a wider distribution than alkaline phosphatase. Neither enzyme stained the cartilage in the primary lamellae. Alkaline phosphatase might contribute to ion transport as suggested in killifish chloride cells and blue crab gills. Acid phosphatase indirectly reflects metabolic activity.

TELOMERASE INHIBITION BY UAB-30 RETINOIC ACID AS A POTENTIAL TREATMENT FOR HUMAN BREAST CANCER. Rebecca C. Wylie, Jonathan D. Matlock, Sharla Phipps, Lucy G. Andrews, Trygve O. Tollefsbol Dept. of Biology, Univ. of Ala. at Birmingham, AL 35294.

Breast cancer, one of the leading causes of cancer-related death in women, currently presents limited treatment options due to several factors, including toxicity and lack of sustained drug potency. Inhibition of the telomerase enzyme has offered new hope in treatment of breast tumors with a decreased likelihood of adverse side effects. This enzyme, which maintains the telomeric ends of chromosomes, is a critical component in the control of chromosomal stability, cellular senescence, and neoplastic transformation. Although silenced in almost all normal somatic tissue, telomerase is active in over 90% of human carcinomas, including breast cancer. Previous research has shown that inhibition of the telomerase enzyme strongly correlates with many antitumorigenic effects. Retinoic acids (RAs) have been shown in several cancer studies to effectively down-regulate the transcription of hTERT, the catalytic subunit of telomerase, thereby decreasing the enzyme's activity. In this study, a new synthetic retinoid, UAB-30 RA, was compared with the more commonly studied natural RAs, 9-cis and ATRA, in the treatment of MCF-7 human breast cancer cells. Effectiveness of treatment and levels of toxicity were measured by the Telomerase Repeat Amplification Protocol, RT-PCR, soft agar analysis, and Apotag. It was found that the retinoids effectively down-regulated telomerase activity and hTERT mRNA, decreased cellular proliferation and tumorigenicity, and induced apoptosis, with UAB-30 resulting in less overall toxicity compared to the natural RAs.


Previous studies on the population status of S. depressus indicated that its numbers were declining. It is currently listed as threatened under the Endangered Species Act. After two seasons (2002 and 2003) of trapping efforts in the Bankhead National Forest and surrounding areas, the data indicate that populations may not be recovering. In the stream site that has historically held the densest population, 2002 data indicated that the population may have stabilized, but in the subsequent season, atypical weather caused unfavorable trapping conditions and may have produced a false indication of decline. The sex ratio of flattened musk turtles in this site has gone from 2.2:1 (M:F) in 1985 to a 1:1 in the 2002-2003 seasons. Although data indicate that at least some recruitment is occurring in the upper inundation zones of Smith Lake in the Brushy Creek and Sipsey Forks and that populations exist in isolated pockets and coves of the reservoir, the size class distribution is heavily skewed toward the larger size classes. This research was made possible by the support of the U.S.D.A. Forest Service, Alabama Power, The Nature Conservancy of Alabama, and the Birmingham Audubon Society.

A STUDY OF SALT STRESS RESPONSES OF THREE TAXONOMICALLY DIFFERENT PLANT SPECIES. Mijitaba Hamissou and Amora W. Hicks. Biology Department, Jacksonville State University, Jacksonville, AL 36265, USA.

Increasing demands for food quality and quantity are two major causes for over irrigation and over fertilization of agricultural lands. These two factors combined are contributing to increase in soil salinity, creating impediments to plant growth and development. Plants cannot extract water from the soil unless the water potential in the root is less than the water potential in the surrounding soil. Plants growing in saline environments must also cope with the potential toxic effects of N[a.sup.+] ions. Certain plant species are known to cope with soil salinity by synthesizing small molecular weight proteins or by accumulating osmolytes in response to high level of soil salinity. Osmotin, a 26 KD alkaline protein is believed to accumulate in the vacuoles of salt stress tobacco cells. Glycine betaine, an osmolyte, is known to be synthesized by members of the Chenopodiaceae in the chloroplast then transported through the phloem to the growing tissues when grown in saline environment. Despite large published research in salt stress, there still is no single salt tolerance biomarker used in salt stress research. The objective of this study is to investigate some molecular and physiological responses of three plant species exposed to increasing concentrations of NaCl. Plants were grown in potted soils, watered and fertilized for 4 weeks then irrigated with NaCl solutions of concentrations ranging from 100 mM to 400 mM. Preliminary data indicated that soil salinity repress root elongation and caused mild reduction in plant relative water content. Our results indicated a possible alteration of protein synthesis by plants subjected to prolonged salt treatment.

SEX RATIO OF HAWKSBILL SEA TURTLE HATCHLINGS FROM NESTS LAID IN SEYCHELLES FROM 1999-2002. Amber Park and Thane Wibbels, Univ. of Ala. at Birmingham, Birmingham, AL 35294. J.A. Mortimer, S. Roberts, A. Rulie, Seychelles Ministry of the Environment, Victoria, Mahe Republic of the Seychelles.

Temperature-dependent sex determination (TSD) is a process in which the incubation temperature of the egg affects the sex of the hatchling. The endangered hawksbill sea turtle, Eretmochelys imbricata, possesses TSD, which may allow it to produce a variety of sex ratios. Therefore, TSD can affect ecological, reproductive, and conservational aspects of the hawksbill population. Seychelles is a group of islands located about 1,000 miles off the coast of Kenya and hosts a relatively large population of hawksbill sea turtles. The goal of the current study was to estimate hatchling sex ratios of hawksbill sea turtles produced on Curieuse and Bird Island in the Seychelles. Data loggers were used to measure the incubation temperatures of nests, as well as sand temperatures during the 1999, 2000, 2001 and 2002 nesting seasons. Hatchling sex ratios were predicted based on the average incubation temperatures during the middle third of the incubation. The results indicated a female bias for three of the four nesting seasons.

MARINE SPONGES FROM THE ALABAMA COAST: OIL RIGS AS ARTIFICIAL REEFS. Patrick M. Erwin and Robert W. Thacker, Dept. of Biology, Univ. of Ala. at Birmingham, Birmingham, AL 35294.

Marine sponges inhabit oil rigs off the coast of Dauphin Island, Alabama, in the northern Gulf of Mexico. The oil rig pilings function as artificial reefs by providing "islands" of hard substrate for colonization by sessile invertebrates in an otherwise soft-bottomed community. The paucity of hard substrate in these environments suggests intense competition among individuals encrusting on the rigs. This study was designed to characterize the sponge communities on the rigs and to investigate the factors allowing these sponges to be successful in a space-limited environment. A total of 7 rigs were surveyed for sponge diversity. Each rig harbored from 2 to 5 sponge species. Sponge species were identified using standard taxonomic methods and sequence analysis of the entire ITS-2 and partial 28S ribosomal DNA (rDNA) subunits. The bright orange sponge Hymeniacidon heliophila dominated the sponge community and was found in high abundance on all rigs surveyed. A purple sponge belonging to the family Halichondridae was the second most common species, though in much lower abundance, often found growing near large colonies of H. heliophila. Other species found in very low abundance were Chelonaplysilla sp., Haliclona sp., and Cliona viridis. Differences in sponge abundance and diversity were minimal among the rigs visited. To assess the anti-fouling activity in the sponges surveyed, gel plates were constructed using sponge homogenate and housed in small Petri dishes. To investigate the bacterial communities associated with rig sponges, a clone library of 16S rDNA bacteria sequences is being constructed.

ACOUSTIC CAVITATION AND ADVANCED CHEMICAL OXIDANTS AS MECHANISMS TO ERADICATE MARINE BACTERIA AND MACROINVERTEBRATES FROM BALLAST WATER. Meghana Gavand*, Ananthakrishnan Ananthanarayana[n.sup.+], James McClintock*, Charles Amsler*, Robert Peter[s.sup.+], Shirley Clar[k.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. [.sup.++]Penn State Univ. at Harrisburg, PA 17057.

Ballast water is considered to be an important factor contributing to the introduction of invasive marine organisms. Using a model marine bacterium (Vibrio alginolyticus), we first determined the mortality during batch treatment exposure to either a series of acoustic cavitation frequencies [ranging from 800 Hz to 4 kHz] (range = 4-35% mortality), different concentrations of hydrogen peroxide [up to 1000 parts per million (ppm)] (range = 1%-33% mortality), or ozone [up to 1000 ppm] (range = 0.6-29% mortality). Subsequent combinations of acoustics coupled with hydrogen peroxide and ozone resulted in optimized maximal mortality of 42%. Using an invertebrate model, brine shrimp cysts and juveniles (Artemia salina) were then subjected to these predetermined optimized conditions and percent mortality was measured. The incidence of mortality for brine shrimp cysts and juveniles under bacterial-optimized conditions (1.4 and 1.8 kHz, 100 ppm ozone, and 100 ppm hydrogen peroxide) was 80-91% and 97-100%, respectively. Acoustic cavitation combined with chemical oxidants may be an effective method to treat ballast water to reduce invasive marine species. Supported by a grant from the US Fish and Wildlife Service to R. W. Peters, C. D. Amsler, and J. B. McClintock.

PHLOXINE B IS A POTENTIAL PROPHYLACTIC TREATMENT AGAINST CHANNEL CATFISH PRE-INFESTED WITH ICHTHYOPHTHIRIUS MULTIFILIIS. Department of Biology, Jacksonville State University, Jacksonville, AL 36265. Mark Haygood, Benjie Blair, Mark Meade, Charles Olander, and Ashley Ward.

Ichthyophthirius multifiliis ("Ick") is a ciliated, parasitic protistan that commonly infests freshwater fish. Economically, "Ick" does not cause a major problem; however, secondary diseases usually precede "Ick" infestation resulting in mortality. By eliminating "Ick", other diseases can be prevented as well. Currently, copper sulfate is the most common treatment used in aquaria and pond environments. The toxic effects of this chemical supercede its benefits. These detrimental effects have increased the need for a safe therapeutant that is approved for use in the food fish industry. Studies conducted on phloxine b have illustrated positive results against "Ick" post-infestation and pre-infestation in channel catfish, Ictalurus punctatus. At a phloxine b concentration of 10 ppm, a 70% mean survival rate was observed while at 0 ppm, a 100% mortality rate was observed in post-infestation studies. Similarly, in a pre-infested study performed, a 50% survival rate was noted at a phloxine b concentration of 10 ppm while at 0 ppm, a 0% survival rate was noted. These results demonstrate phloxine b to be a potential prophylactic treatment against Ichthyophthirius multifiliis.

Papers presented at the 81st Annual Meeting

University of Montevallo

Montevallo, AL 35115

March 17-20, 2004
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Date:Apr 1, 2004
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