Biology senior section.

* Bernard, S.M. Department of Biology, Pittsburg State University. Induced Spawning Of Black Crappie (Pomoxis Nigromaculatus) In A Recirculation System. Production of black crappie (Pomoxis nigromaculatus) as a marketable food source requires the development of a methodology to stimulate and control reproduction. Crappie have been successfully reared in ponds and natural environments; however, the least success has been with artificial environments. Work conducted with crappie in recirculation systems is very limited, and almost nonexistent in determining if these fish can spawn in artificial settings. This project will evaluate the techniques used on other fish species to stimulate artificial and aseasonal reproduction in black crappie. The objectives of this project are to: 1) determine if crappie can be stimulated into their reproduction cycle in an artificial environment, and 2) determine if crappie can reproduce aseasonally. Ten specimens were randomly selected from a brood stock accustomed to a recirculation system. After gender was determined, mating pairs were placed in separate tanks in a recirculation system where temperature and photoperiods were adjusted to mimic those occurring naturally. Four mating pairs were injected with 1000 international units of human chorionic gonadotropin. Three days after injection the crappie began to release eggs and milt. The eggs were fertilized, but did not develop offspring due to a fungal infection, which is the current area of work.

* Claxton, J.L., and A.R. Oller, Ph.D. Department of Biology and Earth Science, Central Missouri State University. Isolation Of Oral Microbes By Selective And Differential Media. The oral cavity is home to thousands of different microbial species. Pathogenic bacteria can lead to oral or systemic diseases such as atherosclerosis. Oral flora populations are known to change with age of human subject, due to hormonal changes and antibiotic usage. This study surveyed teeth extracted from people of known age and sex, with the intent of determining antibiotic resistance over age categories (18-39, 40-59, and 60+). The methodology utilized was to identify gram negative (Enterobacteriaceae) and gram positive (Streptococci) bacteria by culturing microbes on brain heart infusion broth (BHI) and BHI containing blood components as initial media. Sabaroud-dextrose (SAB) broth and BHI supplemented with penicillin and streptomycin were used as initial culturing media for yeasts. Initial classification used gram stains, wet mounts, and flagella stains, with further analysis performed on MacConkey, Mannitol Salt, BHI with blood, Peptone Yeast Agar with sorbitol, SAB with penicillin and streptomycin, and Ueda plates. Two plates were made for each tooth (three in each age group at this time), with one plate incubated aerobically and one incubated anaerobically. All isolated organisms were grown in broth culture and cryopreserved for future use, including antibiotic susceptibility testing. Biooxidation tests were used to further identify microbes. This project will lead to the identification of bacteria via molecular techniques for determination of new species. The research was funded by a grant from Central Missouri State University.

* Dygert, J.R., T.W. Valliant, D.J. Dibben and H.M. Krempa. Department of Biology, Benedictine College. Annual Biodiversity Patterns Of Aerial Invertebrates On The Benedictine Bottoms Mitigation Site. Benedictine College Biology Department is monitoring the biodiversity of the Benedictine Bottoms Mitigation Site, including invertebrates inhabiting the area. Aerial invertebrate sampling was conducted from May 1995 to October 2003 along permanently established transects. Sampling included aerial, aquatic, and terrestrial invertebrates. Sticky traps were used for aerial collection, set up 1-1.3 meters above the ground with 96 [cm.sup.2] covered with the adhesive Tanglefoot, and left for two days. Since 1995, over 62 thousand aerial invertebrates have been collected, enumerated, and identified to order. In 2000, biomass was incorporated into the aerial collection, of which the size was determined for a sub-sample of the invertebrates trapped. The five most commonly occurring orders are: Diptera, Coleoptera, Thysanoptera, Hymenoptera, and Homoptera. The data analyzed up to this point, using ANOVA have shown that the year, month, and habitat are significant factors in determining the abundance of invertebrates. The aerial invertebrate abundance from 1995 to 1998 has shown an overall downward trend, with a small increase in 1999. The highest abundance occurs in May and June with a decline in abundance leading tip to the first frost in late October.

* Jones, D.M., and L. Sievert, Department of Biological Science, Emporia State University. Temperature And Digestive Rate Of Cornsnakes, Elapheguttata. Temperature is a factor in controlling the digestive rate of reptiles. We compared the transit rates of juvenile cornsnakes, Elaphe guttata, at 22 C and 32 C. Snakes were acclimated to the test temperature for at least three days prior to the experiments. Before the experiment they were fed every other day. Thirteen snakes were fed three uniquely marked meals and the time from ingestion to defecation was recorded. The average mass for the snakes was 53.37 g ([+ or -]0.8889 SD), and the meal was approximately 5% of the snake's mass. Transit rates were significantly taster at 32 C than at 22 C (P=0.005319). We used randomized unpaired testing between these two temperatures, at 22 C (5.65 d) 32 C (3.28 d), only using values from snakes which consumed 2 or 3 meals. During our study some of the snakes underwent ecdysis. Transit rate was significantly slower in snakes undergoing ecdysis (P= 0.01701). We again used randomized unpaired testing finding the average of non-shed snakes (5.13 d) and shed (8.82 d), ignoring temperature. We observed that snakes retained feces until they shed.

* Keane, K.J. and A.R. Oller, Ph.D. Department of Biology and Earth Science, Central Missouri State University. Visualizing Human Phenotypic Characteristics Via Molecular Techniques For Forensic Science Applications. Identification of a perpetrator or victim can be difficult to determine when only residual evidence is present. The DNA sample quantity is often too small to run analytical tests, resulting in a large database of DNA profiles without a direction to pursue. While there are currently published PCR protocols for each trait under investigation, this study developed a single PCR reaction to limit the amount of DNA required for thorough genomic analysis. The University Human Subjects Committee approved teeth collection from a local dentist. Age, sex, and other characteristics of the person were provided. Dental pulp DNA was isolated and purified using standard protocols. Blood residue on the teeth was also collected and saved for DNA isolation and comparison to the tooth DNA results. Primers were synthesized for the AMEL gene for sex determination and ABO blood typing genes. Results were visualized and analyzed by electrophoresis on 2% agarose gels. Gels were photographed using a documentation camera. Selected DNA sequences will be excised from the gel and sent for sequencing to confirm correct primer binding. Other sequences such as telomere length for age determination will be included in this study. This PCR technique will allow law enforcement officials to use key identifying characteristics to narrow their search for a perpetrator or victim. This project was funded by a Willard North Award and CMSU.

* Stokes, K.M., L.A. Hilger, D.E. Bowen, M.P. Simon, and J.W. Davis, Department of Biology, Benedictine College. Plant Species On The Benedictine Bottoms Fish And Wildlife Mitigation Site. In 1993, the U.S. Army Corps of Engineers purchased 855 hectares of land located 1.5 kilometers northeast of Atchison, KANSAS in the southern section of the Rushville Bend. The area is now known as the Benedictine Bottoms Fish and Wildlife Mitigation Site (Bottoms). Since 1994, the U.S. Army Corps of Engineers has planted 176,100 tree and shrub seedlings on 550 acres. They have also planted 750 acres with native grass species and legumes. In 1994, the Biology Department of Benedictine College i2nitiated the Benedictine Bottoms Missouri River Biodiversity Assessment Program on the Bottoms. The transition of the Benedictine Bottoms from agricultural land back to its native floodplain habitat provides an opportunity for a long-term biodiversity survey of the Bottoms in order to monitor the success of the Corps' program. The plant biodiversity assessment project began in 1995. All vascular plant species were collected and voucher specimens were made. They are preserved in the Benedictine College Herbarium. These specimens were identified using the Flora of the Great Plains, and updated using the plants. usda.gov website. As new species are collected, they are added to the computer database. This total plant list is continually growing, and currently 263 species, 162 genera, and 61 families have been found on the Bottoms. The process of collecting all plant species is repeated every five years. The first five-year collection was taken from 1995 to 2000. Currently, we are in the second five-year collection from 2001 to 2005. These five-year groups are compared to determine plant succession and changes in plant biodiversity on the Bottoms.

* Valliant, T.W., J.R. Dygert, D.J. Dibben, and H.M. Krempa. Biology Department, Benedictine College. Annual Biodiversity Patterns Of Terrestrial Invertebrates On The Benedictine Mitigation Site. In 1993 the U.S. Army Corps of Engineers initiated a mitigation project to restore 855 hectares of farmland back to its original Lower Missouri River floodplain habitat. Benedictine College Biology Department is monitoring the biodiversity of the Benedictine Bottoms Mitigation Site. Invertebrate sampling on the Bottoms was initiated in May 1995, along randomly chosen 200-400 m transects. Sticky traps were used to monitor the aerial invertebrate population. Pitfall traps were introduced in 1998 to monitor the terrestrial invertebrate population. These 179 [cm.sup.2] pitfall traps were set in the ground and filled with 2.5 cm of preservative to collect terrestrial invertebrates. Specimens were preserved in 75% ethyl alcohol until identification. Since 1998, nearly 10,000 invertebrates have been collected, measured and identified to 21 different orders using this method. The two most common orders are Coleoptera and Orthoptera. These data have been used to determine which factors such as site, year, month, rainfall, temperature, technique, and habitat are important determinants of patterns biodiversity of biomass trends. For example, the amount of monthly rainfall has shown a significant impact on the abundance of these orders.

Rex M. Strange

Southeast Missouri State University