Fall applications are due November 10 each year and spring applications are due March 20 each year.

The LSSU Undergraduate Research Committee announces a call for undergraduate student research proposals. The application window for fall is from August 30 – November 10. The application window for spring is January 30 – March 20.  Awards of up to $500 to support senior capstone project/research-based equipment, supplies, and travel will be granted on a competitive basis. Please Note that food expenses cannot be supported by the grant. Students should consult with their faculty mentor and read the application guidelines. Submitting an application is the final step. Contact one of the URC members if you have questions.

Undergraduate research is an integral part of the student experience at Lake Superior State University. In fact, the University mission and vision statement emphasizes LSSU’s role in helping students develop their full potential, as well as contributing to the growth, dissemination and application of knowledge.

The Undergraduate Research Committee (URC) is charged with promoting and recognizing the student research efforts that occur across campus, as well as encouraging new efforts. The committee consists of faculty members from various academic disciplines.

One of the main roles of the URC is to oversee the Undergraduate Research Fund, a program sponsored by the LSSU Foundation that awards grants of up to $500 to students to support project or research-based equipment, supplies, and travel.

The student grants are awarded once each semester on a competitive basis. To apply for a research grant, students must fill out the online application form.

Guidelines for Application

  1. Have your faculty mentor approve your project description.
  2. Complete the online application
  3. Application section describing your project. A timeline should include specific details. You should be careful about using technical jargon as the review committee is made up of a broad audience. Your project description should speak to the goals/rationale of your project, as well as the methodology. Citations should be included, where appropriate.
  4. Budget section justifying your request for funds (not to be used for wages or food for the researcher). Be sure to include how the rest of the budget will be paid if your budget is greater than $500.
  • Request a letter of support from your faculty project advisor/mentor
  • If your project involves the use of humans or vertebrate animals, please submit your proposal to the appropriate LSSU committee.
    1. Vertebrates: Institutional Animal Care and Use Committee (IACUC)
    2. Humans: Institutional Review Board (IRB)
  • Students may apply for multiple URC grants, but preference will be given to first-time applicants.

Expectations for Successful Award Recipients

  • Upon award notification, recipients are encouraged to write a letter thanking the donor.
  • Funds will be reimbursed only with appropriate receipts or documentation. Students should submit all receipts to their department for reimbursement. For internal transfers, it is recommended that when a student is awarded the money, the amount would be transferred to the department at the time of the award so that they could reimburse the student directly when the purchases are made and the receipts are turned in.
  • A final report is due four semesters after award receipt or before graduation, whichever comes first. Yearly progress reports are also expected.

The Undergraduate Research Committee Members (URC)

Sandi King, DNP, Professor, School of Nursing, Chair of Undergraduate Research Committee
Janine Murray, Administrative Assistant, Lukenda School of Business, Undergraduate Research Committee Assistant

Barbara Evans, PhD, Professor, School of Science and Medicine
Kat Rocheford, PhD, Assistant Professor, School of Natural Resources and the Environment
Gerald Root, Associate Professor, Lukenda School of Business
Russell Searight, PhD, Professor, School of Kinesiology and Behavioral Sciences
Joseph Susi, PhD, Professor, School of Kinesiology and Behavioral Sciences
Jason Swedene, PhD, Professor, School of Arts and Letters
Masoud Zarepoor, PhD, Assistant Professor, School of Engineering and Technology

The Foundation

The funding for the URC student grant fund is made available by donations to the LSSU Foundation Office. If you are interested in making a donation or have questions about donations, please contact Sharon Dorrity, the Director of Constituent Relations.

Grant Recipients and Their Research

Fall 2020 Grant Awardees

Bobbi Jo Caskey

Impact of Plant Species on the Microbial Community of an Aquaponic System

Aquaculture, Hydroponics, and Aquaponics Aquaculture is the term used to describe the breeding, raising, and harvesting of living organisms in an aquatic environment (US Department of Commerce, 2019). “Fish farming” is often a common term. A common example of this is recirculating aquaculture system (RAS). While systems may vary, the concept remains the same. The water that holds the fish circulates out along with any waste or leftover food. It is then brought through a filtration, then an aeration unit that will return oxygen to the water and remove carbon dioxide (Losordo, et. al., 1999). Fish are not the only organism that can be raised as algae, seaweed, and other aquatic plants can also be grown. Hydroponics is a similar concept, however terrestrial plants are produced. It includes growing crops in water rather than soil (Tripp, 2014). Fertilizers, including vitamin and minerals are added to the water to ensure the plants have what is needed for growth. While both hydroponics and aquaponics provide significant advantages, specifically when addressing overconsumption of our food supply, recent studies have shown that a combination of the two could provide an even greater benefit (Junge et al., 2017). Aquaponics, not to be confused with aquaculture, integrates the recirculation of water and aeration of a RAS but uses the terrestrial plants and nitrifying bacteria from hydroponics to rid the system of waste products harmful to the fish. The fish will produce nitrogen waste in the form of ammonia. This ammonia is then broken down into nitrite and then nitrate by the microbes. A buildup of ammonia or nitrite can be harmful to the fish however nitrate is relatively harmless. Nitrate is also the preferred source of nitrogen for plants (Rakocy, et al., n.d). In the end the plant, fish, and bacteria all benefit. Microbes in the System While all components of the system are important, this study will look specifically at the microorganisms. Hydroponics, like aquaponics, often use nitrifying bacteria to produce nitrate, so it can be assumed that the microbiota will be similar, however it is unclear on how the aquaculture aspect will affect these microbes. It is not unlikely to find plant growth-promoting rhizobacteria (PGPR) in a hydroponic system. Rhizobacteria are the bacteria that grow directly on the edges of the roots. This means that they have direct contact with the roots. They maintain a symbiotic relationship and help promote plant growth accomplishing one of the following: reducing nitrogen into nitrate, enhancing stress resistance, stabilizing phosphate and potassium intake (Cakmakci, Donmez, and Erdoan, 2007). While they are extremely common in soil, PGPRs are also used as a biofertilizer to help plant growth in both hydroponics and soil grown crops. It was specifically found that Bacillus spp. Gliocladium spp., Trichoderma spp., and Pseudomonas spp. were common bacteria and can be found in a healthy hydroponic system (Lee, and Lee, 2015). All the previous species are also considered Rhizobacteria. Each species benefits the plant differently. For example, one distinction of Bacillus spp is that some species lowered the infection rate of some pathogens (Nihorimbere et al., 2011). In addition, specific species were reported to increase water quality and growth rate (Nihorimbere et al., 2011). Some species of Pseudomonas spp. were shown to decrease root rot and have some antifungal properties on certain species (McCullagh, et al., 1996). Now turning the attention to the aquaculture side, one study found that two main types of bacteria were found: autotrophic and heterotrophic bacteria. Many autotrophic bacteria were ammonia oxidizers such as Nitrosococcus, Nitrosospira, and Nitrosomonas or nitrate oxidizers such as Nitrobacter and Nitrospira (Rurangwa and Verdegem, 2015). These oxidizers help reduce the amount of ammonia and nitrite (that are harmful to the fish) into nitrate or nitrogen gas that are much less toxic. The heterotrophic bacteria were found to be different depending on the species of fish. For example one study using carp found Alphaproteobacteria and Betaproteobacteria. Later the same group used goldfish instead and found a larger variety of microbes that included Actinobacteria, Bacilli, Gammaproteobacteria, Planctomycetacia, Sphingobacteria, Hyphomicrobium denitrificans, Rhodovulum euryhalinum and Nitrospira moscoviensis (Sugita, et al., 2005). Advantages of Aquaponics/Implications and Relevance Food security is becoming an increasingly relevant topic globally. The United Nations Food and Agriculture Organization (FAO) estimated that by 2050, the global population will reach 9.1 billion and our agricultural system allone will not be able to supply enough food (Sohngen, 2017). In addition, studies have shown a great decline in labor in agriculture (Christiaensen, Rutledge, and Taylor, 2020). Finally, as the amount of people going into agriculture decreases, it was also found that those going into the agricultural sector have less knowledge than previous generations (Kuiper, Shutes, Van Mejil, et al., 2019). Without the proper knowledge and understanding, a series of trial and errors will occur that could have been prevented. Now that we are aware of the problem, it is now time to come up with a solution. Aquaponics, as previously mentioned, recirculation of water along with the aeration of a RAS but uses the terrestrial plants and nitrifying bacteria from hydroponics to rid the system waste products harmful to the fish. The end result would be both a supply of fish and produce grown from the plants. Aquaponics also has a multitude of other benefits. Some agricultural policies such as pesticides, fertilizers, and over use of soil have shown negative effects on the environment (Goudie and Viles, 2003). Aquaponics recirculates the water and only produces organic waste that is easily broken down (Konig, et al., 2016). This makes aquaponics an eco-friendly solution. As well as being an environmentally friendly solution, it is also economically friendly. The largest cost of maintaining an aquaponic system in a commercial style system was fish food (Konig, et al., 2016). The aeration unit, bacteria, and plants contribute to make the water reusable for the fish meaning the cost of water significantly decreases. Finally, aquaponic brings forth the idea of having a local food source rather than having dependence on food being brought in. This dependence can be seen currently in the pandemic. Studies have shown that because of travel restrictions, some consumers are no longer able to visit their typical spot nor were their typical products available (Laguna, et. al, 2020). It is also predicted to see an 17 million increase in food insecure Americans in 2020 compared to 2018 (Gundersen, et al., 2020). If used accordingly aquaponics could help reduce the dependence on transported goods. Aquaponics could be one part of solving the problem of food insecurity. It has shown to have great promise in multiple aspects economically, environmentally, and even more so in food sustainability. However, its full potential cannot be reached without a thorough understanding of the system. Abundant research has been done on the microbial ecosystem of hydroponics and aquaculture, but little has been done in aquaponics. Traditional aquaculture does not typically look into the microbes of the system. One of the main reasons being that DNA sequencing is very expensive to be proforme. In 2015, it was estimated that a whole human genome sequence would cost approximately $4,000 (Wetterstrand, 2020). This study will contribute to the knowledge on the microbial ecosystem. The objective of this study is to identify common microbes of an aquaponic system and to see if the type of plant has any effect on the microbes within the systems. The hypothesis is that while there are some microbes that are predicted to be the same, there will be a significant difference between the systems with different types of plants. Methods: Overview of setup Seven (7) five-gallon tanks will be used in this study. 3 tanks will have plant A (most likely basil), and 3 tanks will have plant B (most likely swiss chard). The remaining 2 tanks will be a control and hold additional fish respectively. These fish will be used as a replacement if one of the fish in the trial system passes on. All tanks will be cleaned with 20% bleach and thoroughly washed out with deionized water. They will then be air dried for 7 days to ensure that any left over chlorine is degraded. The figure below depicts a larger version of the system that will be implemented in this study. It will use a cylindrical tubing cut to look like a “C” to hold the plants grown in rock wool. A submersible water pump will be used to force water into the tubing. The tubing will be held at an angle to allow water to flow back into the tank. To speed up the growth of bacteria, nitrifying bacteria will be added as an additive (this is common in aquaculture). Three goldfish will be placed in each trial tank along with the control. The fish will be fed 1.2 grams of food. Measurements of temperature, ammonia, nitrite, nitrite, pH, chlorine, water hardness, and dissolved oxygen will be taken every Monday, Wednesday, and Friday before the addition of food to the system. This will be done to record and understand the health of the system while the microbiota is being established. This system will be allowed to run for 45-60 days. After the allotted time, microbial samples will be collected, and analyzed. This system was designed by a group of New Jersey students for the Lunar Plant Growth Challenge put on by NASA. Image Credit: Atlantic County Institute of Technology (Smith, 2009) Microbial Analysis In order to identify the microbes, the following procedure will be performed. A sample of water from each tank will be collected in a sterilized beaker. During transportation a clean tin foil cover will be used to prevent contamination. The samples will then be immediately filtered using a filter membrane with .45 micrometer pores. This is large enough to catch any organism, yet still be able to filter out any waste. The DNA will then be extracted from the filter by using an Illustra Bacteria GenomicPrep Mini Spin Kit. Once extracted, PCR will then be used along with a 16s microbiome kit that will help enrich and amplify the bacterial DNA. After this, DNA Clean & Concentrator™ Kits, Zymo Research will then be used to purify the bacteria and ensure that any leftover material from the PCR. Finally, DNA sequencing will be able to be performed and the data will then be analyzed to identify the species of microbes Data Analysis Once the microbes are identified, a series of two group t-tests will be performed. Since the data will not be numbers, some modifications will have to be made. To begin, each system with plant A will be paired with a system form with plant B. In all there will be 3 pairs. Then, all the found microbes will be placed on a spreadsheet. At this point, the microbes will be translated into 1 or 0 depending if the microbe was found in the system (1), or not found in the system (0). Each pair will then have a two group T-test run on them to see if there is any significant difference between the two. After the calculations are completed, confidence intervals will be used and a conclusion will be made. Timeline: Fall 2020: (current) prepare proposal, research, trial systems, present proposal Spring 2021: continue trial research (determining if this is the setup I want to us), practice jjjjjjjjjjjjjjjjjjj maintaining system (to ensure less problem during experimentation) Summer 2021: Work and earn money for project Fall 2021: Perform project, gather and organize data, begin analyzing results Spring 2022: take Bio 499, begin final report, senior presentation Budget: Fish: .25$ each $2 6 in one test strips: 100 for 15$ $15 Submersible water pumps with tubing $84 5-gallon tank 8 x 10 $80 DNA Clean & Concentrator™ Kits, Zymo Research present in lab 16s Microbial DNA prep kit present in lab PCR reagents $~50 ZymoBiomics DNA miniprep kit $274 Extraction filter present in lab Various glassware present in lab PCR machine present in lab Plastic petri dishes (40) $22 Nutrient Agar powder $46.30 Flow cell-NGS Nanopore $100 Overall price: $673.30 Any suggestions would be extremely beneficial, and I thank you for your time. Literature Cited: Cakmakci, R. Donmez, M. and Erdoan, Ü. 2007. The Effect of Plant Growth Promoting Rhizobacteria on Barley Seedling Growth, Nutrient Uptake, Some Soil Properties, and Bacterial Counts. Turkish Journal of Agriculture and Forestry 31:189-199. Christiaensen, L. Rutledge, Z. and Taylor, J. E. 2020) Viewpoint: The future of work in agri-food. Food Policy, 101963. Goudie, A. and Viles, H. 2003. The earth transformed: An introduction to human impacts on the environment. John Wiley & Sons, New Jersey, USA Gundersen, C. Hake, M. Dewey, A. et al. 2020. Food Insecurity during COVID ‐19. Applied Economic Perspectives and Policy 00, 00: 1-9 Junge, R. König, B. Villarroel, M. Komives, T. et al. 2017. Strategic Points in Aquaponics. Water. MDPI. 9(3): 10.3390/w9030182. Konig, B. Junge, R. Bittsanszky, A. et al. 2016. On the sustainability of aquaponics. Ecocycles 2(1), 26-32. Kuiper, M. Shutes, L. Van Mejil, H. et al. 2019. Labor supply assumptions – A missing link in food security projections. Global Food Security 25. Laguna, L. Fiszman, S. Puerta, P. et al. 2020. The impact of COVID-19 lockdown on food priorities. Results from a preliminary study using social media and an online survey with Spanish consumers. Food Quality and Preference 86, 104028. Lee, S. and Lee, J. 2015. Beneficial bacteria and fungi in hydroponic systems: Types and characteristics of hydroponic food production methods. Scientia Horticulture 195: 206-215. Losordo, T. Masser, M. and Rakocy, J. 1999. Recirculating Aquaculture Tank Production Systems A Review of Component Options. SRAC Publication No. 453. McCullagh, M. Utkhede, R. Menzies, J.G. et al. 1996. Evaluation of plant growth-promoting rhizobacteria for biological control of pythium root rot of cucumbers grown in rockwool and effects on yield. Eur J Plant Pathol 102: 747–755. Nihorimbere, V. Cawoy, H. Seyer, A. et al. 2011. Impact of rhizosphere factors on cyclic lipopeptide signature from the plant beneficial strain Bacillus amyloliquefaciensS499. FEMS Microbiology Ecology 79(1): 176-191. Rakocy, J. Masser, M. and Losordo, T. (n.d). Recirculating Aquaculture Tank Production Systems: Aquaponics—Integrating Fish and Plant Culture. SRAC-454. Rurangwa, E. and Verdegem, M. C. 2015. Microorganisms in recirculating aquaculture systems and their management. Reviews in Aquaculture 7(2): 117-130. Smith, H. 2009. Aquaponics. Retrieved November 01, 2020 from Sohngen, T. 2017. The World May Run Out of Food in the Next Decade: Study. Retrieved October 15, 2020 . Sugita, H. Nakamura, H. and Shimada, T. 2005. Microbial communities associated with filter materials in recirculating aquaculture systems of freshwater fish. Aquaculture 243(1-4): 403-409. Tripp, T. 2014. Hydroponics advantages and disadvantages. Pros and Cons of Having a Hydroponic Garden. Speedy Publishing LLC, Delaware, USA. US Department of Commerce. 2019. What is aquaculture? Retrieved October 06, 2020 from Wetterstrand, K. 2020. The Cost of Sequencing a Human Genome. Retrieved November 10, 2020.

Rachel Farina

Immunoglobulin Specificity Testing of Salmonid Species Using Rabbit, Rat, and Chicken Antiserum

Immunoglobulin specificity testing of salmonid species using rabbit, rat, and chicken antiserum Rachel Farina Lake Superior State University School of Science and Medicine Dr. Jun Li Fall 2020 Introduction In the Great Lakes region, several species of salmonid fish are naturally common in the wild and in hatcheries for research, breeding, and human consumption: lake trout (Salvelinus namaycush), rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta), and atlantic salmon (Salmo salar). Immunological fish health is a very important topic discussed, especially in places where fishing is a main industry. Since these species of fish live in such large numbered communities, pathogens can spread rather quickly. Large amounts of disease, illness, and death in fish can economically harm the fishing industry greatly and reduce a common source of food for the human population living in this area (Magnadottir, 2010). As the fish industry increases, the immune system of these salmonid species must be studied fully, and laboratory reagents and practices must be developed further. Most organisms have an immune system, and fish are the first group of organisms with both innate and adaptive immunity (Magnadottir, 2010). The immune system of any organism is composed of many different types of cells and proteins that protect the organism from pathogenic threats. The salmonid immune system depends on the innate immune system heavily for health, much more than adaptive immunity (Uribe, 2011). Complement, a series of soluble proteins, and neutrophils, cells with phagocytic capabilities, are some of the main innate defenses that are present in fish mucosal membranes to fight off and kill pathogenic organisms. The innate system does not have any long-term memory functions, but it can fight off any pathogens very quickly without needing specific antigen interactions (Ma, 2020). Usually, an organism gets infected with a pathogen and is never aware because the innate immune system kills the pathogen before it can become symptomatic, and thus the adaptive immune system is never activated. Salmonids do also have an adaptive immunological response that is long lasting, but slow to activate. Fish have B-lymphocytes and T-lymphocytes both, but no lymph nodes for all the cells to congregate and accelerate the production of antibodies (Magnadottir, 2010). A single antibody is a Y-shaped molecule with variable regions that allows different molecules to interact with a very specific antigen. B-cells in fish secrete antibodies in response to specific antigens from potential pathogens. Then, the antibodies find the antigens in the body and attach to them so other immune cells know what to attack (Owen et al., 2009). It is important to note, especially in the Great Lakes area, that the secretion of antibodies is temperature-dependent and the activity is lowered in cold temperatures (Dixon). IgM antibodies are the most present isotype in salmonids, but they are tetrameters, four Y-shaped units attached together, instead of pentameters, five Y-shaped units attached together. The pentameter IgM is found in humans and mammals (Magadan, 2016). The adaptive immune system needs to be activated by the innate system, so an organism would only produce antibodies to something that the innate immune system could not handle by itself. The adaptive immune system contains a memory system which prevents the fish from getting sick from the same pathogen twice and a faster immune response to familiar pathogens (Owen et al., 2009). Because of this, even healthy fish contain antibodies in their serum. Enzyme Linked Immunoassay Assay (ELISA) testing is a major technique in immunological laboratories. It tests for the presence of a specific antigen or antibody by using the opposite as a reagent. Antibodies are very specific and match with only one antigen with great specificity. The original serum and a known laboratory reagent are mixed together into a well and allowed to incubate so the antigen and the antibodies can bind together. In some cases, a secondary detection antibody is necessary, which is “a[n] antibody that binds a primary antibody that is not enzyme-conjugated” (Alhajj, 2020). This is the indirect ELISA test. Then, a color changing substrate is added, usually horseradish peroxidase (HRP), so the wells can be analyzed in a plate reader (Owen et al., 2009). The concentration of antibodies detected can be determined from a created standard curve using the absorbance readings. Antiglobulin testing is a procedure used in microbiological, virological, and immunological laboratories. It is used to detect the presence of antibodies in one species by using the antiserum of another species. Purified antibodies from the original serum that is being analyzed are injected into a secondary species, where the animal’s immune system creates antibodies against the purified antibodies that were injected (Harmening, 2019). These antibodies can be used as the secondary detection antibodies in an indirect ELISA test (Alhajj, 2020). This can be used to detect the presence of a specific antibody in a laboratory setting and the minimum dilution that detects the antibodies. In this specific study, IgM antibodies were purified from the salmonid species, and anti-fishIgM IgG antibodies were created for the rabbit and rat, and anti-fishIgM IgY antibodies were created for the chicken. This study will test the specificity of each antibody-antiglobulin reaction between the four salmonid species and the three different antiglobulin producing species. Serum already collected from each fish species will be used for all specimens, and all of the antiserum is already created and is stored. These samples are from previous staff and students of Lake Superior State University. The study will determine which antisera works best to detect IgM antibodies for each salmonid species and the minimum dilution at which they are detected. Methods and Analysis The detection method to determine the specificity of each combination of specimen and antiserum will be done by ELISA testing. The salmonid IgM antibodies will be introduced into a well plate, and incubated for 15 minutes at room temperature to allow them to coat the walls of each well. Then, the different antisera secondary antibodies will be introduced to the wells, and allowed to incubate for 15 minutes at 37 degrees Celsius. Last, substrate will be added to each well, incubated at room temperature for 5 minutes, and observed for color change results. In between each step, the wells will be washed with 1x PBS three times to ensure accurate results. The plates will be spectrophotometrically analyzed by a plate reader at a 450 nm to test for specificity (Alhajj, 2020). I will use 96-well plates for analysis. Row A and B will contain control materials, with fetal bovine serum (FBS) as a negative control and Rainbow Trout serum as a positive control. The other columns will be replicates of each fish serum-antisera combination. Each has a series of dilutions, repeated four times: 1:500, 1:1000, 1:2000, 1:4000, 1:8000, 1:16000. This setup can be seen below in Figure 1. The goal of this, is not only to see which antisera can detect the fish antibodies, but also to see the minimum concentration needed for detection. Creating laboratory antisera and isolating the antibodies for clinical use are both time consuming and expensive techniques. It is very resourceful to know what the minimum concentration needed is, so lab reagents can last longer and be used more. I plan to use an ANOVA test to determine if there are any significant differences to prove one antisera species binds more specifically to the fish antibodies than the others. Since there are two independent variables, a two-way ANOVA test will have to be performed. The specificity will be determined for all interactions, with two groups, or factors: fish serum species and antiserum species. There are four levels for the fish serum, and three levels for the antiserum species. This means that degrees of freedom is 3 for the fish serum and 2 for the antiserum species. Table 1: Treatment Groups Lake Trout Rainbow Trout Brown Trout Atlantic Salmon Rabbit Rat Chicken Each will have the exact same amount of replicates, four, and the averages determined by the plate reader will fill in the table listed above. A main effect test, interaction test, within variation test, and an F-test will be run using a two-way ANOVA test to determine significance of this study. If there are significant results, this project may develop farther. References Alhajj, M., Farhana, A. (2020). Enzyme Linked Immunosorbent Assay (ELISA). Statpearls. Dixon, B. The immune response in fish: A brief review. Received on 04 October 2020 from https://www.vin.com/apputil/content/defaultadv1.aspx?pId=11257&id=3863675&print=1 Harmening, D.M. (2019). Modern Blood Banking & Transfusion Practices. F.A. Davis, p. 103-110. Ma, S. (2020). Effects of environmental stressors on the innate function of Atlantic Salmon (Salmo salar). Lake Superior State Senior Project. Magadan, S., Sunyer, O.J., Boudinot, P. (2016). Unique features of fish immune repertoires: particularities of adaptive immunity within the largest group of vertebrates. Results Probl Cell Differ. 57: 235–264. Magnadottir, B. (2010). Immunological control of fish diseases. Marine biotechnology, 12, p. 361–379. Owen, J., Punt, J., Stranford, S. (2009). Immunology. The Journal of Experimental Medicine, p. 660-663. Uribe, C., Folch, H., Enriquez, R., Moran, G. (2011). Innate and adaptive immunity in teleost fish: a review. Veterinarni Medicina, 56, (10): 486–503.

Veronica Shull

Team Superior Acoustics and Geophysical Solutions: A Method of Oil Detection in Bodies of Water Covered by Ice

Team Superior Acoustics and Geophysical Solutions is a senior research group consisting of three Mechanical Engineering students: ________; and one Electrical Engineering student: ____. The students in this group have taken coursework in Acoustics, Signal Processing, Vibrations and Noise Control, Research Methods and Electromagnetics to prepare for the research to be pursued in this project. The team intends to perform experiments to test a method of oil detection in bodies of water covered by ice with the purpose of detecting and mapping oil spills to assist in expediting the clean up process. There is little research available in this area and due to the close proximity of Lake Superior State University to Enbridge’s Line 5, a 4.5 mile long pipeline carrying oil through the Straits of Mackinac, this issue is of greater interest to students at the university and the wider community. The proposed method involves hydroacoustics and uses reverberation time to detect changes from a pristine state. The hypothesis of the method states that the rate, at which a sound field decays changes, based on environmental factors, and the effects of oil on the sound field are measurable. Testing of the method by this team will be done in a scaled tank environment and efforts will be made to implement the method in a full-scale environment. The team will also improve the scaled tank environment by obtaining a material that would adequately represent the acoustic properties of ice without requiring the generation of natural ice, permitting testing to continue when the outdoor temperature isn’t low enough for growing ice by exposure. An impulsive sound source will also be identified by this team to be used in both the scaled tank environment and in full-scale implementation. In order to mitigate issues encountered by future groups with full scale implementation, this team will also collect data from lake environments in order to analyze the ambient noise present to establish a sound floor and investigate potential passive sources. In lieu of using crude oil in a lake environment for testing, an acoustics software package will be obtained in order to perform calculations that will further validate the method. Documentation of decisions made, supporting research and processing code will be compiled for future students to continue validation and implementation of the method. This documentation will be presented at an academic conference and a paper will be published to contribute to the larger body of research in oil detection methods. The team has a budget of $1000 from the School of Engineering and Technology at Lake Superior State University which will be used to obtain materials to test as an ice substitute, research papers, make repairs to the small-scaled tank environment and make improvements on the current sound source used in the small-scale tank environment.

Carlie Weaver

The Effects of PFAS on the Beneficial Gut Bacteria Lactobacillus Acidophilus

Perfluoroalkyl substance (PFAS) contamination has recently become a large topic of discussion (and action) across the nation, specifically in the state of Michigan. These chemicals have incited executive orders, action response teams, and state-declared states of emergency (MPART, 2019). PFAS are long, fluorinated carbon chains that were commonly used in industry from the 1950s until discontinuation in the United States around 2015 as food packaging, nonstick coating, firefighting foam, and an important component of teflon. During these years, these chemicals have found their way into ground water sources, and from there into the human body (Buck et al., 2011). Water sources have been tested and have been found to have much higher concentrations of PFAS, particularly perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS), than is considered safe. Measurements of greater than 30,000 parts per trillion (ppt) have been found in groundwater despite the legal cap for safety being placed at a mere 70 ppt (Reade et al., 2019). Recently, the Center for Disease Control has named PFAS “chemicals of concern” toward human health, as research has shown that they may be one of the causes of numerous diseases, including cancer, diabetes, infertility, and weakened immune system. They may also contribute to changes in the liver, thyroid, pancreas, and more, as the chemicals have a long half-life in the body and are present in serum.(Anderson-Mahoney et al., 2008) (Center for Disease Control, 2018). While PFOA and PFOS have been discontinued in the US, a new chemical has taken its place. Named “GenX,” 2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)propanoic acid is now being manufactured to compensate for the loss of PFOA and PFOS. This chemical has been advertised as safe, despite causing symptoms of disease in lab rats (Shea, 2018). The creator company, Chemours, has endorsed and produced the chemical. Recent tests show that GenX is also present in drinking water and accumulates in the blood (North Carolina Department of Environmental Quality). The exact mechanism of the diseases caused by PFAS is still unknown. Research has yet to disclose all of the effects these chemicals may have on the body, and how they are caused. One potential mechanism of disease may be through disruption of beneficial bacteria in the gut. The human body is known to have a large variety of flora, particularly consisting of a variety of bacteria, that have been shown to play a crucial role in human health, and damage to the microbiome has been linked to cancer, liver disease, diabetes, and more (Jandhyala et al., 2015) (Sekirov et al., 2010). Lactobacillus acidophilus is a species of bacteria found in the human gastrointestinal tract. This species has been shown to have major beneficial roles in human health: Lactobacillus has been shown to protect against liver disease, and increased numbers of the bacterium are also related to a lowered risk of colorectal cancer (Ritze et al., 2014) (Moore and Moore, 1995) (Guarner and Malagelada, 2003). Colorectal cancer is the third leading cause of cancer deaths in the United States, and is affecting ever increasing amounts of young people in the United States (Siegel, Jemal, and Ward, 2009). Because of this connection, killing these beneficial bacteria could be detrimental to the overall health of the US population (Zhong, Zhang, and Covas, 2014). Despite the small amounts of current literature on the subject, PFAS have been shown to disrupt the membrane bilayer of bacteria, and at high enough concentrations have been shown to damage and even kill the bacteria (Fitzgerald et al., 2018). Because PFAS are found in water, have been ingested by humans, and are even found in the blood, it is entirely possible that the presence of PFAS in the GI tract can interfere with or even kill bacteria in the gut, including the beneficial Lactobacillus. The death of Lactobacillus bacteria may thus be a causal factor in diseases such as colorectal cancer and liver disease. The purpose of this experiment is to investigate the relationship between PFAS and human health by determining the effect of PFAS on the beneficial bacteria of the gut. Three PFAS will be tested- the commonly found PFOA and PFOS, and the new GenX chemical. It is hypothesized that all PFAS tested, including the “safe” GenX, will have some inhibitory effects on Lactobacillus acidophilus, demonstrating a possible mechanism of disease in the human body. Methods: The effect of PFAS on Lactobacillus acidophilus was tested in two ways. One of these tests measured the zones of inhibition created by treating Lactobacillus acidophilus with multiple concentrations of PFAS. When a petri dish of bacteria is treated with a small disc covered in an inhibitory chemical and allowed to incubate, a circle of no bacterial growth will appear around the disc- this area is measured and is called a zone of inhibition. The larger a zone of inhibition is, the stronger the inhibitory effect was. The other test performed was a spectrophotometric assay that allowed for the growth of the bacteria to be measured. Each bacterial species has a characteristic “growth curve” that can be graphed when measuring absorbance of a bacterial sample over a long period of time. If the bacteria is treated with an inhibitory chemical, this characteristic growth curve will change. Escherichia coli was used as a control, as it has demonstrated sensitivity to PFAS in previous research. When compared, the data will show if Lactobacillus acidophilus may be even more sensitive to PFAS than E. coli. Zones of inhibition: 50 Petri dishes were inoculated with either Lactobacillus acidophilus or Eschericia coli. Diffusing discs were placed on the center of each plate, and each of the discs was treated with 40ul of either a high PFOA concentration, low PFOA concentration, high PFOS concentration, low PFOS concentration, high GenX concentration, low GenX concentration, or no PFAS at all. This was repeated in triplicate, so that 3 of each treatment group for each bacteria could be measured. These petri dishes were incubated at 37 degrees celsius for four nights, and any zones of inhibition were measured. Spectrophotometric assay: Six 50 mL tubes of liquid growth media were treated with either a high PFOA concentration, low PFOA concentration, high PFOS concentration, low PFOS concentration, high GenX concentration, or low GenX concentration. A 4 mL aliquot was removed from each of the larger stocks and transferred into a separate 5 mL test tube, for a total of 6 tubes. One additional tube of 4 mL untreated liquid growth media was also prepared. Into each of these tubes, 100 ul Lactobacillus acidophilus culture was placed. This process was repeated two more times, for triplicate measurements. The above procedure was followed, but instead of placing 100 ul Lactobacillus into the tubes, 50 ul E. coli was used instead, to allow for a control to compare Lactobacillus to. This created a total of 42 tubes. All 42 tubes were placed into a warm water bath at 37 degrees celsius, and shaken overnight. Beginning the next morning, the absorbance levels of the tubes were measured using a spectrophotometer at 660 nanometers wavelength. This was performed approximately each hour for 8 hours. The absorbance data was used to create a growth curve for the normal bacteria and each of the different PFAS groups. Data analysis: An ANOVA will be performed on the zone of inhibition data to determine which PFAS had the most effect, at what concentration, and which bacteria was most affected. The average growth curve for each treatment group will be plotted and compared to the “normal” growth curve to determine the effects of different concentrations of PFOA, PFOS, and Gen X on the bacteria over time.

Rebecca Weipert

Does Change in Location Affect the Frequency or Severity of Allergies?

The objective of my study is to determine whether there is a correlation between individuals that have moved locations from childhood and adulthood and have increased sensitivity to allergens. Allergies are caused by the immune system’s response to allergens. Allergens are one type of antigen which promote antibody production in the body. Allergens promote a specific antibody to produce, Immunoglobin E (IgE) antibodies. In the presence of IgE antibodies, allergy symptoms occur. These allergy symptoms are also classified as Type I Hypersensitivity reactions. In this study, the allergens that are being focused on are substances that are both foreign and harmless to the body and cause symptoms of hay fever. Hay fever symptoms include any combination of the following: runny/stuffy nose, itchy/watery eyes, inflammation of airway, headache, fatigue and phlegm. The most common types of allergens to cause these symptoms are pollen, dust, mold and dander. Mast cells, basophils and eosinophils are all types of white blood cells that are activated from the production of IgE antibodies. Increased numbers of eosinophils correlate with the presence of allergies; therefore, you are able to count eosinophils in blood samples and detect the presence or absence of allergies. A study conducted in 2012 showed that children living in urban cities had a higher prevalence of allergic diseases than children who lived in rural towns and villages. The children living in the urban areas also reported that they had experienced more frequent and severe allergies than the children living in the rural areas. To conduct this study, a minimum of thirty LSSU student participants will be needed. The participants will be assigned into groups according to their childhood residences. Half of the participants will be in a group with their residence being Sault Ste. Marie (SSM) and currently reside there. The other half will be in another group with their childhood residences from a location other than SSM but live there currently. This groups’ childhood residences must not be within 115-mile radius of SSM. Questionnaires will be provided to each participant pertaining to past hometown information and frequency and/or severity of allergy symptoms since being in SSM. Finger pricks and HemoCue white blood cell analyzer will be carried out and used in order to obtain blood eosinophil counts in each participant.

Sydney Zuke

Examining the Correlation Between Variations of the ACTN3 Gene and Athletic Predisposition in Division II Student-Athletes at Lake Superior State University

In the scientific community, it has been widely accepted that elite athletic performance is a result of a combination of environmental factors and genetic makeup (Georgiades, 2017). Although elite athletic performance is dependent upon many different genes, this research will examine a specific gene, alpha-actinin-3 (ACTN3), also known as the “sprinter” or “speed” gene, and its genotypes (Pickering & Kiely, 2017; Del Cosa et al., 2019). This gene is of particular interest because it encodes the alpha-actinin-3 protein which is responsible for generating fast-twitch muscle fibers. Fast-twitch muscle fibers are responsible for generating high amounts of force quickly. Therefore, the variant form of the ACTN3 gene is common in elite athletes that participate in sports that require explosive power, such as sprinting (Ostrander, Huson & Ostrander, 2019). The ACTN3 genotype will be examined to determine its presence in Division II student-athletes at Lake Superior State University in order to understand if those student-athletes were predisposed to athletic success because of the ACTN3 gene or if their success was determined primarily through effort. Thus, we are examining whether nature predisposes Division II athletes the same as witnessed in elite athletes, or if nurture and training play a larger role in success. This study will take samples from athletes of both anaerobic (power and speed) prevalent sport student-athletes and aerobic (endurance) prevalent sport student-athletes to determine the degree of prevalence of the ACTN3 genotype. The information will be used to determine if there is a correlation between sport type and the presence of the ACTN 3 genotype in this population. This will be accomplished through the collection of DNA samples through cheek swabbing and subsequent analysis. Norman et al. (2014) conducted similar methodology. Purpose Statement: The purpose of this research study is to determine if there is a significant correlation between the presence of ACTN3 genotype and athletic predisposition in Division II student-athletes at Lake Superior State University. Relation to field of study: Nature, or genetic predisposition, has been proven to be a precursor to athletic success at the elite level (Pickering & Kiely, 2017) As the competition becomes stronger, the athlete must have nurtured their athletic talents, but they must also have similar genetic predispositions in order to truly compete (Ostrander, Huson & Ostrander, 2019). Thus, we see athletes selecting one sport to compete in at the elite level. This is not true at lower levels. At the high school level, it is common for athletes to play 2, 3 or even 4 sports (Bell et al., 2016). Some of these athletes may not play sports at the collegiate level. However, for those that do, they must rise to a new level of competition. It is not known if the level of competition at the Division II level is ‘elite’ enough in which genetics must play a role in order for athletic success. Thus, this study adds to the field of kinesiology by beginning to answer the question regarding nature versus nurture at the Division II level of competition. The answer to this question has much to offer in terms of training methodology and recruitment of student-athletes. Methodology: Upon approval of the HSIRB committee, student-athletes who participate in Division II athletics at Lake Superior State University will be asked to participate on a volunteer basis. Following the informed consent process, student-athletes from anaerobic and aerobic based sports (tennis, basketball, volleyball, cross-country, track and field and golf) will be asked to set up a time for DNA collection through swabs. No procedures will be performed until informed consent is provided to the participants. Student-athletes will meet the primary and faculty researcher in the genetics lab and complete a short demographic survey to identify gender and sport. Numbers will be used in place of identifying information. Following the demographic survey, the primary researcher will conduct a cheek saliva DNA swab. Following the DNA swab, student-athlete participation in this study will be complete. The primary researcher will immediately prepare the swab for DNA analysis to determine the presence of the ACTN3 genotype.

Fall 2019 Grant Awardees

Brendan Lukomski

Jared Emmack URG Award

Comparing rhizospheric differences between young growth and old growth Eastern White  Pine (Pinus strobus)

In this study the goal will be to determine differences in rhizospheric soil microbial communities between differing aged stands of Eastern White Pine (Pinus strobus).  Soil samples have been collected at 3 locations; a site near Tahquamenon State Park, a location just southwest of Sault Ste. Marie, MI, and a site near Copper Harbor, MI.  The extracted microbial DNA from the soil samples are then analyzed for microbial community abundance as well as diversity within the community.  This will determine whether or not the Eastern white pine individuals are selecting certain microbial community characteristics throughout their lifetime.

Brianna Regan

Haven Barna URG AwardeeThe impacts of enrichment on the problem-solving abilities of rats (Rattus norvegicus)

The physical, psychological, and overall well-being of animals can be negatively impacted by captivity. Enrichment is a method of keeping captive species engaged by giving them activities to partake in throughout the day. Enrichment devices could include toys, novelty objects, food, sensory activities, and much more.  My project will utilize rats to investigate the impacts enrichment can have on problem-solving ability. There will be a control and a treatment group. The rats in the control group will not be exposed to enrichment, while the rats in the treatment group will. Each rat will be evaluated by recording the time it takes them to complete a maze before and after the experiment takes place. This data will be analyzed to look for differences between the rats that were exposed to enrichment and the rats that were not to assess how problem-solving ability is impacted by enrichment.[CP_CALCULATED_FIELDS_RESULT]

Lucas Sheppard

Nash Johnston URG Award

“Effects of salinity on serum cortisol in Atlantic salmon”

This project will look into safe handling techniques of Atlantic salmon (Salmo salar) in regard to performing a “Dip” into a salt bath for vaccinations. Salmon will be randomly assigned into three salt test groups with a further level of variation being length of test. Blood was taken post different salinity exposure in order to determine a safe concentration of salt to use for a “Dip” as well as length of time. Total blood cell count and plasma cortisol levels were analyzed under microscope and via ELISA test respectively. ANOVA test was used for data analysis to find a difference between the tests: serum cortisol and salinity treatment

Victoria Size

Renee Resendes URG AwardThe Effects of PFAS on the Development and Mitochondrial Function in Zebrafish Ova

Renée Resendes of Windsor, ON tested the effects of PFAS (Per- and Polyfluoroalkyl
Substances) on the development and mitochondrial function of zebrafish ova. The two most
common forms of PFAS are PFOA and PFOS, which are persistent in the environment and of
common exposure to humans. The study aimed to determine if there was a correlation between
changes in embryonic development and energy production, as a result of exposure to PFOA,
PFOS, and in combination. Her results showed that PFOA and PFOS in combination affected
embryonic development, caused changes in energy regulation, and within these conditions even
resulted in embryo death before 14 days old. Research of PFAS is of value because of the
potential harm these chemicals have on reproductive and developmental processes and overall
adverse human health effects.

Jessica Thompson

Taylor Severance URG Award

Stress levels related to Acetaminophen usage in social conformity

Recently, some newly found side effects of acetaminophen, the active ingredient in Tylenol, have shown suppressed emotional output. This study was designed to use social conformity as a medium to monitor any behavior, mood, or physical changes acetaminophen may have on those who consume it. A total of 48 participants were evaluated during a small group activity on their cortisol levels, mood states, blood pressure/pulse, and behavioral changes. The data gained will be analyzed to determine if acetaminophen is decreasing physiological stress reaction and mood changes, and if the suppression of stress may result in a behavioral change.

Spring 2020 Grant Awardees

Kassidy Hadix

Casey URC2019

I will be looking for the frequency of two common gene mutations in the Eastern Upper Peninsula Caucasian population. The genes of interest are Factor V (responsible for blood clotting) and methylenetetrahydrofolate reductase (MTHFR, which regulates homocysteine buildup). These mutations are important because they can lead to serious conditions, such as abnormal blood clots and miscarriages.  DNA samples will be obtained from non-invasive cheek swabs; amplified using PCR and quantified using the new gene fragment analyzer in the School of Science and Medicine at LSSU. We will compare the EUP’s prevalence of these mutations to that of the overall U.S. population. Knowing the frequency of these two mutations can help to educate healthcare professionals and the general public about the risks associated with these conditions, so that people can better regulate their health.

Breanna Kemppainen

Chemical cleaners are used to clean surfaces in households, hospitals, and public places to protect us from illness. Most cleaners use a spray-then-wipe method but it is unclear how the physical action of wiping contributes to removing bacteria. Some cleaners may not be actually killing bacteria, but friction from wiping may be enough to physically remove them. In this study we will clarify the contribution that friction makes to cleaning surfaces by counting the number of bacteria removed from a surface when using dry materials versus the amount removed when using the same material wet with water or a cleaner.

Sophie McConkey

Elias URC2019

The intent of this research project is to enable a mobile robot to accept human assistance for the purpose of deviating from its autonomously generated path, thus reaching its desired destination, while avoiding obstructions. The results of this research will enable Automated Guided Vehicles to overcome the challenge of navigation within dynamic environments. The team plans to use a TurtleBot 3 Waffle Pi, equipped with a Color Sensor Distance Measuring Sensor, and Gyro Sensor, an 8 Megapixel camera, and a Nvidia Jetson TX2 computer board to accomplish this task.

Rebekka Ranta

Harnden URC2019

Identifying and understanding the effect that perceived stress and resilience levels have on an individual’s performance when presented with an acute-stress inducing task is very important.  For university students, many are frequently faced with negative situations that are not always directly related to academics. By identifying such, the way an individual performs under pressure can be better understood. Participants were evaluated on their stress and resilience perceptions, their performance on the stress-inducing task, and their heart rate levels. Research surrounding this topic is of value because it provides information that could improve the health, well-being, and retention of university students.

George Schaffer

Mills URC2019

I am researching how lung function may influence symptoms of panic, such as shortness of breath, a racing heartbeat, feeling faint, or shaking. I measured lung function using a spirometer, a small handheld device that measures exhalation speed and lung capacity. I also asked participants to fill out a demographic survey and a panic survey detailing their history of panic attacks, the specific symptoms they had if they’d experienced a panic attack, and whether or not they had experienced a panic attack recently.