In 2012, a project involving plant DNA barcoding was started at Gordon State College to improve student engagement in upper division and senior level research courses. DNA barcoding involves in collection of specimens of interest for an identification and laboratory analysis of DNA barcode sequences. The data are then placed in a database for subsequent analyses. One of the most important components of the Barcode Initiative is the construction of a public reference library of species identifiers which could be used to assign unknown specimens to known species. This provides an excellent molecular tool for students to be trained to collect, manage, and analyze DNA barcode data. In Fall 2012 and Spring 2013 by the Biotechnology and special research topics classes, three groups of six students isolated genomic DNA from 30 vegetable and 30 fruit species from local grocery stores and the nine plant species from the Gordon State College Walking Trail. Genomic DNA preparation was done using PureLinkTM Plant DNA Total DNA Purification Kits (Invitrogen, Carlsbad, CA). The students amplified chloroplast rbcL sequences by polymerase chain reaction (PCR) using those isolated the genomic DNA templates and DNA sequencing was done by GeneWiz. The students currently enrolled in the special research topics course in Fall 2013 are analyzing DNA sequences using MEGA software to understand the evolutionary relationship of these species and further analysis DNA barcoding data. This provides an excellent educational research project within the undergraduate upper division biology lab classes and the special research topics course.
Biology Dept, School of Arts and Sciences, Gordon State College, Barnesville, GA
P132 • Donna A. Goodenow, Patricia A. Koplas, Jeffrey Thomas
Why Did You Die? A Post-Mortem Analysis of Avian Tissues
Animals may be brought into rehabilitation centers because of physical trauma, toxic exposure or disease. When dead animals are recovered, the cause of death is not always apparent. Tissue samples can be used in association with veterinary necropsies to assess potential causes of mortality. The main objective of this project is to optimize the staining procedures for raptor tissue in order for an avian pathologist to be able to determine any potential pathology in the tissues. Staining protocols for the Hemotoxylin and Eosin, the Hopps Brown gram staining, and the Giemsa stain have been modified to suit various raptor tissues including but not limited to gut, pancreas, kidney, liver, heart and stomach. All raptor tissue was provided by the Carolina Raptor Center and the tissues were embedded in paraffin, sliced and set by a local histologist with the Cannon Research Center. These modified staining protocols have been successful and the quality of the stains confirmed by experts in the field. With histological analysis of these raptor tissues it is possible to diagnose problems in the raptor population. The data from this analysis, combined with the medical data from the Carolina Raptor Center, can provide detailed information about both the natural and anthropogenic causes of disease and injury in natural populations of birds of prey.
Longterm Chemotaxic Ability of Caenorhabditis elegans Following Treatment With 6-Hydroxydopamine
Parkinson’s disease, a neurodegenerative disorder, is caused by the degeneration of dopaminergic neurons in the substantia nigra. The neuronal death could be due to oxygen free-radicals. Superoxide dismutase is responsible for converting the oxygen free-radicals into less toxic by-products. Characteristics of the disease include disturbed movement, bradykinesia in arms and legs, and resting tremors. It is believed that the neurotoxin 6-hydroxydopamine (6-OHDA) is the cause of this degeneration. Caenorhabditis elegans (C. elegans) are an ideal organism to study this effect because the species has a simple nervous system. Evaluating how this neurotoxin works on the C.elegans gathers information that relates to the human condition of Parkinson’s disease, and this may provide progress towards a cure. Past research has shown that treating the nematode with 6-OHDA causes a negative effect in the organism’s ability to chemotax. The chemotaxis assay provides information on the organism’s speed and movement. In this experiment, the C. elegans were treated with 6-OHDA and tested for three successive days. An ANOVA test compared the worms’ abilities to chemotax, based on whether they were treated with 6-OHDA or not. This experiment evaluates the difference in the organism’s response when the resting period following treatment is extended to twenty-four, forty-eight, or seventy-two hours.
Belmont University
P134 • Shannon Theobald, Marlee B. Marsh
Using Mab M24-2 (Α Fish Lysozyme) to Examine the Host-Parasite Relationships in Livers of Fish From Saluda Shoals Park, Columbia, South Carolina
Fish innate immune responses are routinely evaluated as indicators of immune function and status following exposure to pathogens, biological response modifiers, immunotoxicants, and nutritional regimes. Recently, we developed monoclonal antibody (mAb) M24-2 that recognizes lysozyme in several species of fish (E.g. Fundulus heteroclitus, F. grandis, Ictalurus punctatus, Morone saxatilus) used in comparative immunological studies. Lysozyme is found in macrophages and neutrophils and is one of several humoral and cellular factors associated with innate immunity in all vertebrates. The purpose of this study was to use mAb M24-2 to examine cellular profiles of immune cells in livers of parasite-infected and uninfected livers of 41 fish from Saluda Shoals Park in Columbia, SC. There is no current model for fish immune responses to parasites. Furthermore, any direct role(s) of neutrophils and macrophages in fish immune responses to parasites has yet to be determined. This is mostly due to a lack of suitable reagents. In this study, we probe paraffin-embedded livers of several species of freshwater fish ((E.g. Lepomis spp., Ictalurus spp. and Micropterus spp.) and compare the lysozyme profiles of parasite-infected vs. uninfected individuals.
