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Common Bean (Phaseolus vulgaris) provides protein, complex carbohydrates, and other valuable nutrients for more than 300 million people in the tropics and around the world. Common bean is particularly known for its ability to provide fiber, with some studies showing that it aids in the lowering of blood sugar. It is also used as a model species for more complex plant species, as it is easy to grow and make genetic crosses for studying gene linkage. In this project, the goal was to identify if there was a significant difference in the quality and quantity of DNA isolated from fresh versus frozen leaf tissue. Fresh, young, trifoliate leaves were directly collected from newly grown common bean plants for DNA isolation using CTAB (Cetyltrimethylammonium bromide) buffer to help break down leaf tissue. The same procedure was used with common bean leaf tissue that had been stored in the -80 degrees deep freezer for more than five years. Differences in color, texture, and ease of grinding the leaf tissues were some physiological comparisons identified between fresh and frozen tissue. Both isolated DNA were run on a gel electrophoresis to visualize results. Of two fresh and frozen samples each, the results indicate that the frozen samples were more concentrated by visualization as well as quantification with a Nanodrop. Ultimately, the frozen tissue provided higher concentrations of DNA. Keywords: Common bean, DNA, CTAB, Gel Electrophoresis, Nanodrop Acknowledgements: We acknowledge USDA NIFA Capacity Building Grant Awards# 2016-06642 and 2016-06476 for funding my internship and the project as part of the student experiential learning goal and the Project Director, Dr. Gulnihal Ozbay for providing this internship opportunity.

DSU Summer Research Symposium 2021

Does Prolonged Storage of Plant Tissue have a Negative Effect on DNA Quality?

Welcome to Delaware State University’s Annual Summer Research Symposium, and thank you for attending! Our symposium represents the culmination of students’ summer experiences in research at Delaware State University. This event showcases the work of college students working on faculty-led research projects, as well as middle and high school students experiencing research for the first time working in research groups. Whatever the case, this summer has been a unique experience for the students, and we hope a transformative one. Even for students who are not planning a career in scientific research, their summer experience has given them skills they will draw on for years to come. 
 Please join us on the mainstage and in Gather for the live event, and visit the recorded poster presentations any time.
 The student participants and organizers of the various summer programs at DSU would like to thank all of the research mentors who guided our summer program participants. There were many DSU faculty, graduate students, and other research and program staff who gave their time and energy to help our students have a fun and educational summer experience. Without their generosity and patience these summer programs would not be possible.
 For the research participants, we hope that you will maintain contact with your research mentors and colleagues. Science is a relatively small community made up of disciplinary “neighborhoods”. If you continue to work in a discipline, the other folks in that discipline will be like your neighbors that you run into regularly. Staying in contact with them will help you keep up with the news of the neighborhood, and open doors to educational and career opportunities. Whether you continue in science or take a different career path, for this summer you have been part of the world of research, and you will benefit from that perspective for the rest of your life.

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Welcome to Delaware State University’s Annual Summer Research Symposium, and thank you for attending!

poster

Toxoplasma gondii is a parasite that infects most warm-blooded organisms, including humans and causes the infectious disease, Toxoplasmosis. Toxoplasma gondii is transmitted to humans through consumption of contaminated raw or undercooked meats. While it is transmitted to grazing animals through consumption of feed and/or water contaminated with T. gondii in cat feces. Toxoplasmosis can cause humans and animals alike to have miscarriage, stillbirth, congenital defects, and other clinical symptoms. Currently, there is no widespread vaccine or treatment for T. gondii in sheep and goats. Therefore, once identified it is necessary that preventative mechanisms are taken. Hence, the objective of this study was to test for T. gondii in lambs found on Delaware State University’s Hickory Hill farm. To identify T. gondii, blood samples were collected from 15 lambs and the white blood cells separated, isolated, and stored at –80 oC for further processing. After isolation, deoxyribonucleic acid (DNA) was extracted from the white blood cells of each animal and used to conduct polymerase chain reaction (PCR) using a surface antigen one (SAG-1) gene specific forward and reverse primer to amplify the T. gondii gene. After PCR amplification, gel electrophoresis was conducted to fractionate the gene of interest by size and allow for visualization. Data indicated that none of the 15 lambs tested were positive for T. gondii at Hickory Hill Farm as the SAG-1 gene was not present. Under the conditions of this study, it can be concluded that T. gondii parasite is not present on the farm.

Identification of Toxoplasma gondii in Katahdin Lambs

Identifying the Eastern Oyster Growing Season in Rehoboth Bay, Delaware Jannah Zerrad1, Aaron Bland2, and and Gulnihal Ozbay2, Ph.D. Caesar Rodney High School, Camden Wyoming, Delaware1 Department of Agriculture and Natural Resources, Delaware State University, Dover Delaware2 Eastern Oysters, Crassostrea virginia, are valued for their culinary and ecosystem functions, such as improving water quality and providing habitat. Wild oysters have recently struggled to maintain their population in Rehoboth Bay, Delaware due to overharvesting and disease. However, oyster aquaculture farmers have recently begun to grow oysters in Rehoboth Bay, selling to restaurants and households. In addition, artificial oyster reefs were made in Rehoboth bay to increase the wild Eastern Oyster population. The observations of water quality are essential for identifying which dates are the best or worst in the given sites for oysters to survive and grow, both for aquaculture and restoration activities. Water samples were collected from seven sites every two weeks in Rehoboth Bay, Delaware. Measurements of temperature, salinity, and dissolved oxygen were obtained using the YSI 556 multiprobe. Lastly, Chlorophyll a was measured using the Aquafluor Fluorometer. Trendlines for temperature, salinity, dissolved oxygen, and chlorophyll from each of the seven sites were calculated and graphed. We found that the beginning of August-late September were the best times for ideal temperature conditions for Eastern Oysters. Mid-August and later November were found to be good salinity conditions while all sites were suitable conditions for dissolved oxygen in early August-late November. It was also found that all sites were lower in chlorophyll a than expected, suggesting there may be less algae than ideal for oyster growth. The overall best times for growing Eastern Oysters in Rehoboth Bay, Delaware would have been mid-August to late September. These findings may help oyster farmers and conservation managers improve their oyster rearing activities in Rehoboth Bay. Keywords: Dissolved oxygen, Salinity, Chlorophyll, Temperature, Eastern Oyster, Rehoboth Bay Acknowledgments: We acknowledge USDA NIFA Capacity Building Grant Award# 2016-06642 for funding my internship and the project as part of the student experiential learning goal and the Project Director, Dr. Gulnihal Ozbay for providing this internship opportunity.

Identifying the Eastern Oyster Growing Season in Rehoboth Bay, Delaware

Effect of Salinity and Dissolved Oxygen on Blue Crab (Callinectes sapidus) Size in Blackbird Creek, Delaware  Daivik Arora1, Jackie Maina2, and and Gulnihal Ozbay2, Ph.D. 1The Charter School of Wilmington1, Wilmington, DE 2Department of Agriculture and Natural Resources, Delaware State University, Dover, DE Blue crabs are a keystone species in the Delaware bay and are an important source of revenue. However, due to overfishing and water pollution, the crab population has declined significantly, going from 800 million in 1990 to 200 million in 2008 in the Mid-Atlantic region. Water quality changes can be linked to the development of low oxygen areas called dead zones in the feeding areas of the crabs, diminishing their food sources. Higher salinity levels in crab habitat has been associated with larger crabs due to its significance in shell development where calcium carbonate is the main component of the exoskeletons. In this research project, the effect of salinity and dissolved oxygen on crab size was determined using monthly water monitoring data from 2017 and 2020. The study was carried out in Blackbird Creek, Delaware, and 5 sites were identified based on dominant marsh vegetation and salinity gradient. The water quality testing was carried out using the YSI 556 multiprobe sensor and crab sampling was conducted using crab traps and trawling method. 108 crabs were used in this study. Carapace width was determined using calipers. The highest salinity average was recorded in site 1 (5.8 ppt) and the lowest average in site 5 (3.6 ppt). Dissolved oxygen had the highest mean in site 1 (6.6 mg/L) and the lowest mean in site 3 (5.6 mg/L). The largest carapace width average was at site 1 (113.1 mm), while the smallest width average was found in site 4 (94.2 mm). The preliminary results indicate that the largest crabs were from site 1, suggesting a positive relationship between salinity and crab size. These results contribute to the existing knowledge base on salinity and dissolved oxygen influence on blue crabs. Continued monitoring would provide more conclusive insight regarding applicability to blue crab management. Keywords: Dissolved oxygen, Salinity, Blue crab, Carapace, Blackbird Creek Acknowledgements: We acknowledge USDA NIFA Capacity Building Grant Awards# 2016-06642 and 2016-06476 for funding my internship and the project as part of the student experiential learning goal and the Project Director, Dr. Gulnihal Ozbay, and the Project Director, Dr. Shobha Sriharan, respectively for providing this internship opportunity.

Effect of Salinity and Dissolved Oxygen on Blue Crab (Callinectes sapidus) Size in Blackbird Creek, Delaware

Determining the Relationship Between Environmental Factors (Temperature, Turbidity, and Salinity) and Total Vibrio in Water Samples from Slaughter Beach, Delaware Bhini Arora1, DevothaTumushimiyimana2, Rosemary Ebirim3, and Gulnihal Ozbay2,3, Ph.D. 1TheCharter School of Wilmington, Wilmington, DE 2Food Science and Technology Graduate Program, Human Ecology Department, Delaware State University, Dover, DE 3Department of Agriculture and Natural Resources, Delaware State University, Dover, DE Vibrio are a group of comma shaped bacteria, in the family of Vibrionaceae, which are aquatic microorganisms that inhabit salt and brackish waters (salt and freshwater mixed together), and arepathogenic to humans and other sea life. Biological pollutants (such as Vibrio bacteria) that reside in water are considered to be threatening to ecosystems and human life. Measuring water quality parameters such as temperature, turbidity, and salinity helps us determine the correlation between these factors and the levels of Vibrio. The objectives of this research are to evaluate the total vibrio(V. vulnificus, V. tubiashii, V. coralliilyticus, V. parahaemolyticus) in sea water samples collected from Slaughter Beach, DE, and to determine the relationship between environmental factors such as temperature, turbidity, and salinity and the level of Vibrio in the water. Water samples were collected from two different sites (Cedar Creek and Boat ramp) from June to July, 2021 at high and low tides in Slaughter Beach, Delaware. The samples were collected bimonthly. A YSI-556 multiprobe system was used to measure temperature, and salinity, while turbidity was monitored using a Global Water WQ770 turbidimeter. Vibrio were monitored by Thiosulfate Citrate Bile Salts Sucrose (TCBS) and Luria Broth (LB). 100 microliters of seawater were inoculated at 26°C and 37°C. Cedar Creek demonstrated high amounts of Vibrio spp. with high levels of salinity and temperature to both tides from June to July compared to the Boat ramp site. The Boat ramp site demonstrated high turbidity at high and low tides from June to July. This is a continuous project; more research is being conducted to come up with the concrete results and recommendations. Keywords:Vibrio, Temperature, Turbidity, Salinity Acknowledgements: We acknowledge USDA NIFA Capacity Building Grant Award# 2016-06642 for funding my internship and the project as part of the student experiential learning goal and the Project Director, Dr. Gulnihal Ozbay for providing this internship opportunity.

Relationship between Environmental Factors​ (Temperature, Turbidity, Salinity) ​ and Total Vibrio in Slaughter Beach, Delaware

Strawberry (Fragaria × ananassa) is the most cultivated horticultural crop among the small berries due to its rich, healthy components. The major barrier in strawberry cultivation is caused by gray mold fungus (Botrytis cinerea Pers.) that causes an extensive loss in production. The predominant use of copper-based fungicides for controlling the gray mold fungus has been toxic to the plants, and fungal strains were developing resistance. Therefore, the present work aimed to evaluate and identify an alternative approach for controlling the gray mold fungus by screening the strawberry germplasm for resistance. Developing new cultivars requires the effective identification of resistant strawberry genotypes; hence, the current study was aimed at the evaluation of strawberry genetic resources under lab conditions by establishing an invitro leaf disc assay. The leaf discs from six different strawberry varieties were screened on 0.6% agar media and infected with actively growing gray mold fungus mycelial discs and incubated in the growth chambers maintained at 25˚C with 80% of humidity for two days under dark and 5 days under 16/8 hours of light/dark conditions. The fungal infection injury rate was measured at 7 days after infection. We identified two highly resistant, two moderate resistant, and two sensitive genotypes among the six varieties screened. The fungal infection was successful on leaf discs and the resistant genotypes showed less necrotic lesions, and sensitive genotypes showed more lesions. This assay provided a simple and rapid method to identify the strawberry genotype's resistance to gray mold fungus and potential donor genotypes.

Evaluation of Strawberry Germplasm for Gray Mold Fugal Resistance

Blueberries (Vaccinium corymbosum L.) are a very important crop because of their unique health benefits. The major hurdle for growing blueberries is the pH in the soil, as they need acidic soil. Hence improving the pH level tolerance in the blueberries provides a viable option for the growers to cultivate in a wide range of soil types and allow them to grow in several regions of the world. With this, our objective of the study was to screen the blueberry germplasm variation for high pH levels and identifying the tolerant lines. We used an in-vitro plant twig assay to screen the population using phosphate buffer solutions containing several pH gradients ranging from 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, and 7.0. In the assay, we used 6 different varieties of blueberries for testing the variation for pH level. Among six varieties, we screened, identified one highly resistant, 4 moderately resistant genotypes, and one sensitive genotype to the high pH. The resistance lines maintained a stay green nature at the high pH level, but the sensitive lines turned to brownish and chlorophyll degradation was observed. Developing new cultivars for high pH tolerance requires the effective identification of tolerant blueberry genotypes; therefore, the current approach was aimed at evaluating blueberry genetic resources under laboratory conditions by establishing an invitro plant twig assay. This assay provided an easy and time-consuming method to identify the blueberry genotypes tolerant to high pH and a better genetic source for developing the pH tolerant varieties.

Screening of Blueberry Varieties for pH Variation

Changes in the Morphology and Nutrient Content of the Marsh Grass Collected from Different Saline Conditions Nishtha Kumar 1 , Mayavan Subramani 2 , Ph.D., Venugopal (Kal) Kalavacharla 2 , Ph.D. 1 Orange County School of the Arts, Santa Ana, CA, 2 Delaware State University, Department of Agriculture and Natural Resources, Dover, DE Marsh grass (Sporobolus alterniflorus) is essential to marshland formation as it holds sediment in place and provides habitat for a variety of aquatic species. However, removing them from marshlands would cause negative trophic cascades. Additionally, they can also thrive in salinity conditions ranging from high to low. Runoff from farms and other sources of fertilizer pollution is polluting water bodies and causing soil and water concentrations of nitrogen, potassium, and phosphorus to increase. How do salinity and nutrients affect the morphology and nutrient content of marsh grass? To answer this question, we collected saltmarsh grass (Sporobolus alterniflorus). from higher,

Changes in the Morphology and Nutrient Content of the Marsh Grass Collected from Different Saline Conditions

Comparative Analysis of Chromatin Immunoprecipitation in Common Bean and Salt Marsh Grass Vishal Thamaraikannan1, Vasudevan Ayyappan2, Ph.D. and Venu (Kal) Kalavacharla2, Ph.D. 1John P. Stevens High School, Edison, NJ. 2Delaware State University, Department of Agriculture and Natural Resources, Dover, DE Chromatin immunoprecipitation is a technique used to examine interactions occurring within the cell between DNA and its binding proteins. Chromatin is a condensed form of DNA found inside the nucleus of eukaryotic cells. In order to access chromatin, it must be extracted using chromatin isolation. We conducted an experiment with two plants, common bean and salt marsh grass, to compare the reactions of the plants when their chromatin was isolated and how the two genomes differ when studied using gel electrophoresis. Beforehand, samples of each plant were stored in -20OC to slow cell metabolism and keep the cells alive until used. The leaves were ground using liquid nitrogen and mixed with 2mL of nuclear isolation buffer to begin isolation. 1.7mL of 2M glycine was added to halt the cross-linking reaction. After filtration, the tubes were centrifuged at 4000rpm at 4oC for 10 minutes. The resulting DNA pellets were dissolved in a cold nuclear isolation buffer and transferred to 1.5mL tubes with 300µL of 15% Percoll solution and centrifuged again at the same settings. 600µL of nuclear lysis buffer was added and sonication was used to lyse the nuclear envelope. 20µL of the samples collected before sonification and 20µL of the samples after were inserted into a small gel casting for electrophoresis. After centrifuging twice, the marsh grass pellet was larger than the common bean pellet, meaning more DNA sank down because of its greater density in the marsh grass sample. The marsh grass DNA fragments were also more visible than the common bean fragments in the electrophoresis scan. Since the DNA of marsh grass was denser and more visible in electrophoresis than the DNA of the common bean, it is proven that the DNA of the salt marsh grass is larger. Keywords: Chromatin Immunoprecipitation, Chromatin, Eukaryotic Cells, Gel Electrophoresis, Centrifuge, Sonication, Density Acknowledgements: We acknowledge USDA NIFA Capacity Building Grant Awards# 2016-06642 and 2016-06476 for funding my internship and the project as part of the student experiential learning goal and the Project Director, Dr. Gulnihal Ozbay for providing this internship opportunity.

Comparative Analysis of Chromatin Immunoprecipitation in Common Bean and Salt Marsh Grass

Aragonite Saturation as an Indicator for Oyster Habitat Health in Delaware Inland Bays Amin Boukari1, Memory Nakazwe2, and Gulnihal Ozbay2, Ph.D. 1Cesar Rodney High School, Dover, DE 2Delaware State University, Department of Agriculture and Natural Resources, Dover, DE Oyster farming in Delaware is a crucial industry and it makes about $300,000 to $500,000 in sales every year. Oysters use calcium carbonate ions in the form of aragonite and calcite to build their shells. Ocean acidification can lead to a decrease in carbonate ions making building these shells difficult. When the aragonite saturation state falls below 3, calcifying organisms become stressed, and when it drops below 1, their shells begin to dissolve. Therefore, measuring the aragonite saturation state yields crucial insight into the suitability of habitats to support oyster growth. This project aimed to calculate the aragonite saturation state from seven sites in Delaware Inland Bays to determine their feasib

Aragonite Saturation as an Indicator for Oyster Habitat Health in Delaware Inland Bays

Investigating Oyster Health Status by Studying Oyster Condition Index and Natural Oyster Sets in Rehoboth Bay, Delaware Sara Berzinji1, Aaron Bland2, Memory Nakazwe2, Mohana Gadde2, and Gulnihal Ozbay2, Ph.D. 1Ransom Everglades High School, Miami, FL 2Department of Agriculture and Natural Resources, Delaware State University, Dover, DE Over the past few decades, oyster (Crassostrea virginica) populations have been in decline. One of the many bodies of water affected by this is the Rehoboth Bay. This research focuses on assessing the health of oysters placed in seven different sites along the Bay. Assessing which locations are thriving will help in creating more successful artificial reefs and therefore lead to healthier aquatic ecosystems. While there is a large issue with oyster populations, this research focused on those in the Rehoboth Bay. Bags with oysters were deployed at each of the seven sites in the early summer of 2020 and were collected 5-6 weeks later. This process was repeated two more times, in late summer and then in the fall of 2020. Spats were counted and the condition index was calculated by measuring the height, width, and depth of the oysters along with the weight of their shells and tissue dry and wet. Calculating the condition index and counting the number of spats help determine if the oysters were healthy. The condition index remained generally constant throughout the 3 testing periods and the recruitment rates at each site were also constant with the exception of the BBR site which had a higher average number of spat along with the S Cove site. These results can be used to create healthy and successful oyster reefs in the Rehoboth Bay and better the health of the ecosystem. Keywords: oyster restoration, oyster recruitment, condition index, Rehoboth Bay, Crassostrea virginica Acknowledgments: We acknowledge USDA NIFA Capacity Building Grant Award# 2016-06642 for funding my internship and the project as part of the student experiential learning goal and the Project Director, Dr. Gulnihal Ozbay for providing this internship opportunity.

Next from DSU Summer Research Symposium 2021

Identification of Toxoplasma gondii in Katahdin Lambs

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DNA extraction from Common Bean (Phaseolus vulgaris) leaves