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BIOLOGICAL SCIENCES PAPER ABSTRACTS.

REVERSE ENGINEERING WORM MUSCLE USING CRISPR GENE EDITING. RYAN LITTLE FIELD, UNIVERSITY OF SOUTH ALABAMA.

The Caenorhadbitis elegans roundworm is the simplest model animal that has striated muscle. Use CRISPR gene editing techniques, my lab is developing and testing novel molecular biology tools to follow and perturb muscle development and growth in living worms by fluorescence microscopy. By precisely separating the "head" and "tail" domains of muscle myosin isoform A, we found that the myosin "tail" was able to assemble into thick filaments in the absence of the motor function contained in the "head" domain and that the lengths of the actin filaments may have been re-specified to a new length.

OPTIMIZING BODY COMPOSITION IN LABORATORY FED SEA URCHIN LYTECHINUS VARIEGATUS. YUAN YUAN, LAURA HEFLIN, MICKIE POWELL AND STEPHEN WATTS, UNIVERSITY OF ALABAMA AT BIRMINGHAM.

Optimization of formulated sea urchin food could improve nutrient utilization and biochemical composition of the gonad, which could directly affect the quality and economic value of sea urchin roe. Studies have examined the relationship between diet and body composition using single formulated diets which provided no options in dietary content. It is not known if sea urchins can or will preferentially store specific macronutrients or macronutrient ratios when food choices of varying macronutrient content are offered. In this study, individual urchins were offered pairwise moist gel-based diet combinations varying in the proportion of protein and carbohydrate for 43 days. Urchins offered diet combinations generally preferred the diet with most balanced macronutrients (equi-proportioned protein: carbohydrate ratio, i.e., least extreme protein: carbohydrate ratio). High protein diets were correlated with high protein content of gonads (48-61% dry weight). High proportion of carbohydrate diets were correlated with increased carbohydrate content of gonads (9- 20% dry weight). Gonad lipid content ranged from 19-24% dry weight, with the highest lipid levels found among urchins consuming high levels of carbohydrate. These data indicate that sea urchins choose between diets of varying macronutrients and can preferentially store specific macronutrients. In contrast to other organisms, L. variegatus can store protein while limiting carbohydrate storage. This can be an advantage to the organisms living in an environment where protein availability is episodic. We further hypothesize that gonad composition will affect the commercial appeal of roe.

THE EFFECT OF INCUBATION TEMPERATURE ON SEX AND MORPHOLOGY IN A LIZARD. ARIEL STEELE AND DANIEL WARNER, AUBURN UNIVERSITY.

The developmental environment plays a pivotal role in shaping phenotypes and fitness of all organisms. Perhaps the most enigmatic example of environmental effects is the influence of developmental temperature on an individual's sex, a phenomenon known as temperature-dependent sex determination (TSD). The first description of TSD was based on a study conducted 50 years on an African lizard (Agama agama). Although novel at this time of publication, this landmark study consisted of low sample sizes and provided a poor description of the sex-determining reaction norm in this species. Our goal was to revisit this work and better characterize the pattern of TSD in A. agama. In addition, we aimed to quantify the effects of constant and fluctuating incubation temperatures on a variety of fitness-relevant traits of offspring. Eggs were obtained from an invasive population of A. agama in Miami, FL, and randomly assigned to one of nine incubation treatments: six constant temperature treatments and three fluctuating treatments that mimic field conditions. We then measured hatchling morphology (snout-vent length, head size, mass), growth, and sprint performance as indicators of fitness. Size measurements will be continuously taken every six weeks to determine the ontogenetic timing of sexual dimorphism and to determine if sexual dimorphism is influenced by incubation temperature. Preliminary data suggest that warm incubation temperatures produce mostly female offspring. This ongoing research will provide a critical evaluation of the long-term effects of developmental temperature on fitness-relevant traits, and provide insights into the adaptive significance of TSD.

THE EFFECT OF INCUBATION TEMPERATURE ON SEX AND MORPHOLOGY IN A LIZARD. ARIEL STEELE AND DANIEL WARNER, AUBURN UNIVERSITY.

The developmental environment plays a pivotal role in shaping phenotypes and fitness of all organisms. Perhaps the most enigmatic example of environmental effects is the influence of developmental temperature on an individual's sex, a phenomenon known as temperature-dependent sex determination (TSD). The first description of TSD was based on a study conducted 50 years on an African lizard (Agama agama). Although novel at this time of publication, this landmark study consisted of low sample sizes and provided a poor description of the sex-determining reaction norm in this species. Our goal was to revisit this work and better characterize the pattern of TSD in A. agama. In addition, we aimed to quantify the effects of constant and fluctuating incubation temperatures on a variety of fitness-relevant traits of offspring. Eggs were obtained from an invasive population of A. agama in Miami, FL, and randomly assigned to one of nine incubation treatments: six constant temperature treatments and three fluctuating treatments that mimic field conditions. We then measured hatchling morphology (snout-vent length, head size, mass), growth, and sprint performance as indicators of fitness. Size measurements will be continuously taken every six weeks to determine the ontogenetic timing of sexual dimorphism and to determine if sexual dimorphism is influenced by incubation temperature. Preliminary data suggest that warm incubation temperatures produce mostly female offspring. This ongoing research will provide a critical evaluation of the long-term effects of developmental temperature on fitness-relevant traits, and provide insights into the adaptive significance of TSD.

SUBTLE TEMPERATURE DIFFERENCES MAY WELL DETERMINE WHO WINS: A STORY OF FOUR SUBMERGED AQUATIC PLANT SPECIES. MOLLY MILLER, DAVID NELSON AND TIMOTHY SHERMAN, UNIVERSITY OF SOUTH ALABAMA. JAMES MAHAN, 2PLANT STRESS AND WATER CONSERVATION LABORATORY USDA/ARS.

As temperatures increases globally, shifts in the distribution of plant species are expected, with unknown effects on invasive species abundance. It is then of value to understand the role increased temperature may have on invasive species. Although nonhomeothermic organisms are the mercy of environmental temperatures, their physiology is still temperature dependent, with species dependent thermal optima. By identifying the thermal optimum of a species and determining the amount of time spent annually in that optimal temperature zone, success can be predicted under different temperature regimes. Here we identify species-specific differences in the thermal optima of four submerged plants, Ceratophyllum demersum, Hydrilla verticillata, Myriophyllum spicatum, and Vallisneria neotropicalis. Utilizing a biochemical approach, activity of a key metabolic enzyme NADH malate dehydrogenase (MDH) was used to assess the thermal dependencies of Km and Vmax in each species. A Michaelis-Menten model was then employed to predict reaction velocity across a range of temperatures (10 - 40DegC). The predicted reaction velocities were compared to multiyear in situ temperature data. At low temperatures (10 - 20DegC), all three species had similar thermal behavior. However, at temperature > 20DegC, enzyme activity H. verticillata exhibited a sharp increase to a level 2-3 times higher than M. spicatum and V. neotropicalis. H. verticillata is metabolically more competent at lower temperatures (earlier in season) allowing rapid growth earlier than other coexisting species. This data suggests that as water temperatures increase, the highly invasive H. verticillata will be favored over concurring species. Additionally, a northward expansion of the dioecious, southern biotype of this species is likely.

ECOLOGY OF BALD EAGLES (HALIAEETUS LEUCOCEPHALUS) IN ALABAMA. ANDREW COLEMAN, ALABAMA A&M UNIVERSITY.

A knowledge gap exists regarding the ecology of Bald Eagles (Haliaeetus leucocephalus) in Alabama. This species was reintroduced by the Alabama Department of Conservation and Natural Resources in the mid-1980s, and the nesting population currently stands at 100-200 nesting pairs. The present study initiated an examination of nest site selection and predation behavior of Bald Eagles in Alabama. The locations of a collection of eagle nests were compared with USGS land cover data in ArcMap 10. A buffer of one mile was set around each nest location, and the total acreage of each land use category that was within that buffer was summarized. Additionally, many of these nests were surveyed for discarded prey items, and the presence of turtle remains was noted. A variety of turtle species, including Eastern Musk Turtle (Sternotherus odoratus) and Common Map Turtle (Graptemys geographica), were documented on the ground beneath nests. If possible, the carapace length and width of the recovered turtle shells were measured to examine potential size limits of turtle prey that Bald Eagles can seize.

CAPTURING CAVITY-NESTING ANTS WITH ARTIFICIAL NEST TRAP CONSTRUCTED OUT OF PLASTER. ISAAC HEINKEL, GLENN MARVIN AND PAUL DAVISON, UNIVERSITY OF NORTH ALABAMA.

Artificial nest traps (a.k.a. artificial nesting sites, supplemental ant nests) made of various materials including bamboo internodes, plastic tubes, and wood (dowels, lathing, wood blocks) contain preformed cavities which attract entire ant colonies, founding queens, or colony fragments. Such nest traps have been used in studies of colony structure, resource limitations, and ant diversity. Given the common use of plaster of Paris (often augmented with activated powdered charcoal) as a substrate for artificial nests in laboratory studies of ants, springtails, and other arthropods, we tested the efficacy of artificial nest traps constructed out of plaster (Ant-Coops) with and without activated powdered charcoal. Ant-Coop design and construction details will be presented along with ant capture data from 1000 Ant-Coops placed in an oak-hickory upland forest in northwest Alabama. These inexpensive and easily constructed nest traps can be an important new tool to investigate ant ecology (e.g., distribution, abundance, and life history)

SOLVENT EFFECT ON THE EXTRACTION OF ANTIOXIDANTS. WYVOLYN KIRKLAND, JACKSONVILLE STATE UNIVERSITY. NIXON MWEBI, JACKSONVILLE STATE UNIVERISTY.

Numerous fruits and vegetables are believed to be rich in antioxidants. Antioxidants are key in preventing cell damage caused by oxidants found in biological systems. Garlic and red pepper are an excellent source of foods rich in antioxidants. In order to effectively study, quantify, or evaluate antioxidants, one needs to extract them from such foods. A brief literature survey portrays an array of extraction procedures that may at times be confusing. It is therefore imperative to determine the key factors that promote effective extraction of these active biomolecules from vegetables. This forms the basis of our study: to determine the solvent effect on the extraction of antioxidants and polyphenols. We used a spectrophotometric technique that employs the Ferric Reducing Antioxidant Power (FRAP) as a means of measuring the total antioxidants extracted from the vegetables. Our findings indicate that 40% aqueous Acetone, 80% aqueous Ethanol, and 40% aqueous Methanol solvents when used, yield the highest concentration of antioxidants

GEOGRAPHICAL DISTRIBUTION OF ANIMAL-ROAD MORTALITY INCIDENTS IN MOBILE COUNTY, ALABAMA: A PRELIMINARY ANALYSIS. ADAM STERN AND STEPHANIE JETT, UNIVERSITY OF SOUTH ALABAMA.

Our study investigates how roadkill assemblages change across a rural to urban gradient in Mobile County, Alabama and explore factors (e.g. habitat, resource availability, behaviors) that may be inclusive or exclusive to specific species. The current project is one part of a larger project designed to assess specifically scavenger ecology across urban to rural gradient, focusing on factors that shape scavenger assemblages. Roadkill surveys are recognized as a useful tool in assessing the presence of animal species in a study area. The current study will supplement the larger study on scavenger ecology by providing insight into the variety of species who are active in the study area as well as knowledge about human-animal interactions as a result of human encroachment into these species' environments. Currently, there is a sizable body of work that seems to indicate that urbanization may present many obstacles to nonhuman animal species, such as an increase in road density and habitat fragmentation. Select species in some areas may be better adapted to the pressures of urbanization (e.g. better vehicle avoidance methods) and, therefore, be less susceptible to succumbing to animal-road mortality incidents. Preliminary results will be presented discussing differences in roadkill assemblages across the gradient as well as preliminary analyses of patterns in geographical distribution as it pertains to land use and habitat structure.

SEDIMENTARY RECORDS OF RECURRENT PHOSPHATE SPILLS TO A COASTAL ESTUARY. RUTH CARMICHAEL, UNIVERSITY OF SOUTH ALABAMA. JACOB HALL, ELIZABETH HIEB AND PAVEL DIMENS, DAUPHIN ISLAND SEA LAB. ELIZABETH D. CONDON, UNIVERSITY OF NORTH CAROLINA AT WILMINGTON. KIMBERLY CRESSMAN, GRAND BAY NATIONAL ESTUARINE RESEARCH RESERVE.

At least two large phosphate spills are known to have occurred from a fertilizer processing facility to the Grand Bay National Estuarine Research Reserve's Bangs Lake on the northern Gulf of Mexico since 2005. Following these spills, there was a spike in phosphate concentrations (as high as 7 mg/L), a significant drop in pH (to as low as 3.7), and on one occasion, fish and shellfish kills. To define spill periods and determine the fate of phosphorus within the estuary through time, we measured the concentration of phosphorus retained in sediments relative to distance from the spill site. Sediment cores (8 cm diameter X 50 cm long) were compared spatially within Bangs Lake and to cores collected in nearby Bayou Heron, a site more remote from the spill area. Cores were sectioned at 1 cm intervals down to ~24 cm, and each section was analyzed for grain size, particulate organic phosphorus concentration (POP), and porewater phosphate concentration. Core sections also were dated by Lead-210 activity to trace historical phosphorus inputs to each site, and trace element analyses will be used to further corroborate the source of residual phosphorus. We found higher phosphate concentrations in sediments and porewater in Bangs Lake compared to the reference site (while other nutrient concentrations were similar or lower). Peaks in POP concentrations in southeastern Bangs Lake corresponded in time to at least one known major phosphate spill (2005). Although the known source of phosphorus to the estuary is on the western side of Bangs Lake, hydrological processes that flush sediments and nutrients from the Lake may concentrate finer sediments and associated particulate phosphorus in the southeastern part of the Lake. These data provide a spatial and temporal record of phosphorus additions and retention within Bangs Lake to inform companion studies on water quality and primary production as well as future studies of effects on biota.

CITIZEN-SCIENCE PROVIDES DATA ON SEASONAL OCCUPANCY OF WEST INDIAN MANATEES (TRICHECHUS MANATUS) IN THE NORTHCENTRAL GULF OF MEXICO. ELIZABETH HIEB, COURTNEY NELSON-SEELY AND NICOLE TAYLOR, DAUPHIN ISLAND SEA LAB. RUTH H. CARMICHAEL AND ALLEN AVEN, DAUPHIN ISLAND SEA LAB, UNIVERSITY OF SOUTH ALABAMA.

The northcentral Gulf of Mexico was historically considered outside the typical range of the endangered West Indian manatee (Trichechus manatus); however, in recent years reported manatee sightings have increased in this region. To better define the extent of manatee occurrence in the northcentral Gulf of Mexico, we used citizen-science methods, compiling data from opportunistic public sightings of manatees in understudied areas of Alabama and Mississippi. More than 2,000 live manatee sightings were documented from 1978-2015, with peak sightings occurring in rivers and subembayments along the AL-MS coastline during warm months (Jul - Aug). Manatee mortalities, which have significantly increased since the mid-1980s, were most often recorded Nov - Feb and attributed to cold stress. We analyzed the effect of public education and outreach activities on the number and timing of reported sightings to detect potential bias in our use of citizen-science methods. Our results indicated that while targeted outreach efforts were effective in generating manatee sighting reports, the temporal distribution of sightings was primarily driven by manatee presence in local waters. We found that quantitative and consistent documentation of opportunistic, citizen-sourced data enhanced knowledge of manatee habitat use over a broad geographic area and on a decadal time-scale, demonstrating the importance of the northcentral Gulf of Mexico as seasonal manatee habitat. This long-term monitoring effort has increased our understanding of manatee movement ecology and population distribution and is essential to guide effective management and recovery efforts, especially in light of ongoing anthropogenic impacts and large-scale environmental perturbations to local waters.

AGE SPECIFIC PATTERNS IN WINDOW MORTALITY IN AVIAN POPULATIONS. EMMA RHODES, JOEL BORDEN AND JOHN MCCREADIE, UNIVERSITY OF SOUTH ALABAMA.

Building collisions, especially window collisions, pose a major anthropogenic threat to birds. Mortality caused by building collisions is estimated to be between 100 million and 1 billion annually, and it is the second largest source in the U.S. for direct human-caused bird mortality. While several studies have focused on the frequency of building/window collisions in avian populations and the role played by temporal differences in abundance during migration, little research has explored the possible relationship between age and mortality in window strike collisions. If juvenile birds exhibit a pattern of higher mortality from window strikes, it might possibly be attributed to less experienced individuals. However, various other factors need to be taken into account including the species specific ratio of adult to juvenile birds and how these ratios might change seasonally in periods of migration. The primary goal of this study is to determine if there is a direct age-mortality correlation in avian window strikes using a large dataset of curated museum specimens that will classify birds by age based on physiological, morphological, and plumage features. This project is a collaboration of multiple universities and museums in order to examine window strikes on a regional scale and its impacts primarily in the Southeastern U.S. In addition to the lack of research on a possible age correlation with window strikes, there is also a paucity of data from southern states, including Alabama, on window strike mortality and no known study has been done in Alabama examining a correlation with age. Current findings of this project include considering species specific ratios of age groups in avian populations as well as aging avian specimens based on presence/absence of the bursa of Fabricius, which is a key developmental organ currently considered to only be found in juvenile birds. With more increasing anthropogenic threats affecting bird populations, it is vital that preventive measures are taken to prevent unnecessary mortality due to window collisions, which is accomplished by studying the impact of the threat. In conclusion, the goal of this research is to provide significant implications on how window strike mortality affects avian populations and to identify the mechanisms driving window strike mortality for the conservation of avian populations.

MOLECULAR CLONING OF A CDNA ENCODING A SARCO/ENDOPLASMIC RETICULUM CA2+ ATPASE (SERCA) FROM Y-ORGANS OF THE BLUE CRAB (CALLINECTES SAPIDUS) AND ANALYSIS OF SPATIAL AND TEMPORAL PATTERNS OF MRNA ABUNDANCE. MEGAN ROEGNER AND RD WATSON, UNIVERSITY OF ALABAMA AT BIRMINGHAM. HY CHEN, JAPANESE INTERNATIONAL RESEARCH CENTER FOR AGRICULTURAL SCIENCES.

Stage-specific increases in intracellular free Ca[less than or equal to]+ stimulate ecdysteroid production in the molting glands (Y-organs) of crustaceans. Intracellular Ca[less than or equal to]+ levels are regulated by proteins intrinsic to the plasma membrane and membranes of organelles. These include Ca[less than or equal to]+ pumps, e.g., plasma membrane calcium ATPases (PMCAs) and sarco/endoplasmic reticulum calcium ATPases (SERCAs). In order to better understand the role of intracellular calcium signaling in the regulation of ecdysteroidogenesis, we used a PCR based cloning strategy (RT-PCR followed by 3'- and 5'-RACE) to clone a full-length cDNA encoding a putative SERCA protein from the Y-organs of the blue crab (Callinectes sapidus). The cDNA includes a 3060-bp open reading frame that encodes a 1020 amino acid SERCA protein with 80% identity to known crustacean SERCA sequences. Phylogenetic analysis showed the blue crab SERCA protein clusters with other arthropod SERCAs. An assessment of tissue distribution revealed the SERCA transcript was widely distributed across tissues of the crab. SERCA transcript abundance in Y-organs was assessed by quantitative PCR after eyestalk ablation, and during a natural molting cycle. The observed stage-specific changes in SERCA cDNA levels are consistent with the hypothesis that Ca[less than or equal to]+ signaling and intracellular Ca[less than or equal to]+ regulatory proteins play a critical role in the endocrine regulation of crustacean molting.

THERMAL SPIKES CAUSED BY THE URBAN HEAT ISLAND EFFECT RESULT IN DIFFERENTIAL EGG SURVIVAL OF A NON-NATIVE LIZARD (ANOLIS CRISTATELLUS). JOSHUA HALL AND DANIEL WARNER, AUBURN UNIVERSITY.

Embryonic development in ectotherms is very sensitive to abiotic nest conditions. In reptiles, high incubation temperatures often result in relatively short incubation periods and large hatchling size, but extremely high temperatures can result in cardiac arrest and death. Human altered habitats, which potentially create novel thermal conditions in the soil due to the urban heat island effect, may therefore create new selection pressures for developing embryos. The urban heat island effect can increase temperatures in cities as much as 12[infinity] C, and our preliminary data suggests that soil temperatures differ markedly between urban and natural areas in locations where reptiles deposit eggs. We measured the temperatures of potential nest sites of the Puerto Rican Crested Anole (Anolis cristatellus) in both urban and natural areas of Miami-Dade county where this lizard and several other anole species are naturalized. We bred crested anoles in the lab and subjected their eggs to 5 incubation treatments that mimic potential temperature regimes from our field data, three of which included a thermal spike of the way through embryonic development. Preliminary results suggest that thermal spikes increase metabolism and reduce egg survival and that each are a function of the magnitude of the spike. These results suggest that urban environments create novel selection pressures that potentially result in embryonic adaptation to novel temperature regimes or in novel nest-site selection strategies by females.

IMPAIRED SOCIAL BEHAVIOR IN MICE WITH TYROSINEMIA TYPE I IS ASSOCIATED WITH INCREASED MYELINATION OF THE CEREBRAL CORTEX. MARISSA MOORE AND GORDON MACGREGOR, UNIVERSITY OF ALABAMA IN HUNTSVILLE.

Social behavior and cognitive deficits have recently been observed in individuals with the rare disorder tyrosinemia type I, which is caused by an autosomal recessive fumarylacetoacetate hydrolase (FAH) deficiency. Without treatment with NTBC the lack of FAH causes an accumulation of harmful metabolites that leads to severe liver dysfunction within the first years of life. While the current treatment with NTBC has drastically increased the survival rate of tyrosinemia type I patients, it is unable to prevent the neurological impairments affecting them. In order to investigate the recent social behavior deficits, the sociability of mice with tyrosinemia type I treated with NTBC (Fahmut), wild-type mice drinking NTBC (WT-NTBC) and wild-type mice drinking water (WT-Water) was analyzed. The social behavior of mice was investigated using Crawley's three-chambered social test. The buried food test was also conducted in order to analyze mouse olfaction, which is critical for mouse social behavior. Mice brains were extracted and microscopically evaluated for myelin after staining the cerebral cortex with Luxol Fast Blue. Our results show that Fahmut mice spend twice as much time investigating a dummy mouse rather than a novel mouse in comparison to wild-type controls, indicating sociability deficits are caused by the disease and not NTBC treatment. Mice with Fahmut also show abnormal behavior in that they do not spend more time with a novel mouse over a familiar mouse. Tyrosinemia type I mice displayed increased myelination of the cerebral cortex compared to wild-type mice. The increased myelination could create malformed neuronal pathways and synapses that could be a causative factor in the behavioral impairments seen in mice with tyrosinemia type I.

MICROBIAL ART: AN EMERGING SCIENTIFIC INQUIRY MODEL. SARAH ADKINS AND J. JEFFREY MORRIS, UNIVERSITY OF ALABAMA AT BIRMINGHAM.

Microbial art has long been used as a creative outlet for microbiologists and even had a hand in the discovery of the first therapeutic antibiotic. Yet in the school setting many science students do not get to dabble in the creativity that can lead to scientific discoveries. In fact, students often do the opposite by completing "cook-book labs", i.e. prearranged labs that leave little room for curiosity and exploration.

We have proposed a new scientific inquiry model for the microbiology lab classroom at UAB that is anchored on creativity: a petri dish art model. This new curriculum allows students to create personalized living artwork as a platform for scientific inquiry. Over the course of the semester, students isolate colored microbes from natural soil samples, create artwork using these microbes, use standard microbiology techniques to identify their isolates, and finally create personalized experimental designs to answer their own questions about the microbial ecology revealed in their paintings. During this curriculum change, we are collecting data on how effective the curriculum is to a comparable control group on the topics of student engagement, course material, and how students view the role of creativity in science.
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Publication:Journal of the Alabama Academy of Science
Date:Feb 1, 2017
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