Undergraduate Research
Committee

The Effectiveness of Rice as a Cost-Effective Soil Bacteria Capture Method

Soil is an important habitat for organisms witch provide many important functions to natural systems. Multiple studies showing that soil fungi help plants access and take up vital nutrients (Halifu et al, 2019; Chen, et al. 2020; Kang, et al. 2021). These natural functions have been found to transfer over to agricultural systems as well. Wissuwa and other researchers (2020) found that a microbial fungus was found to help rice plants access phosphorus in deprived soils. Pepper plants that were connected to a large network of fungus were found to have a heightened drought tolerance (Marasco, et al. 2012). However, for many decades conventional agricultural practices have been unsustainable. These practices rely heavily on artificial manipulation of the environment to keep the soil productive. The common practice of tilling disturbs soil structure and causes microbial communities to change in population structure (Smith, et al. 2016). Growing the same crop year after year depletes the soil of nutrients, requiring the addition of fertilizers. Growing the same crops also causes a larger population of species that feed off the crop, so farmers need to use pesticides to control pest populations. In the past few years, the idea of organic agriculture has grown in popularity. This method of agriculture looks to rely less on the conventional ways of agriculture, and more on natural proses. As organic agriculture relies less on tillage and chemicals it looks to nature systems for inspiration. That may be plant-based fertilizers, companion planting or many other methods.

One important practice of organic agriculture is the use or beneficial organisms commonly referred to as beneficial. Beneficial provide many functions such as population by bees and butterflies and predation of pests by ground beetles and lady bugs. One subset of beneficials is easily overlooked, microscopic soil organisms . Due to degraded and altered soils soil life can be in a poor condition. Exceptionally if the soil has been under conventional agricultural practices. Tillage disrupts soil structure and causes a change in population of soil microbes (Smith, et al. 2016). So it may be necessary for soil microbes to be introduced to the soil to ether change diversity or reintroduce vital species. It is possible to buy soil microbial starters but there is no guarantee that those microbes are naturalized to a system’s natural prosses. The cost of buying these starters may be inefficient for small producers and individuals gardeners. One possibility is using microbes from a local location. There native and naturalized microbes will be used to natural prosses in the environment. Reddy (2011) described a processes to capture local microbes from an undisturbed habitat. The captured microbes can then be transported and spread upon organic agricultural systems. Reddy’s processes involves leaving a box of boiled white rice in a local field or forest where the local microbes move in to feed upon the rice (2011). This study will use this method of soil microbe capture and determine how this method does in capturing the true diversity within the soil biome. The second objective of this study is if there is any difference between using white rice and a native species of rice. To evaluate these questions four plots will be set up in the fields and open areas of Lake Superior State University’s Hunt Creek Field Station. Each plot will consist of three boxes set upon bare soil. The boxes have thin bottoms with holes drilled in them to give soil microbes access to the inside. Each plot will receive one box of boiled white rice, one box of boiled wild rice and one box of clean potting soil, which will act as a control. A soil sample will be taken from each plot. The boxes will be left in the field for five days. At the end of the time, each box will have samples taken out of them. The samples will be run using the 16s Metagenomics test. This test was used by multiple other studies to gather similar data (Valverde et all, 2016; Sumerta et all, 2020). This will give the percentages and number of species present in each sample. The test from the boxes will be compared to the test results from the soil samples taken. The main information that will be looked for, is how closely a box represents its corresponding soil sample and is there any statistically significant difference in the values between the boxes and the natural soil among the plots.

Design and Experimentation of a High Accuracy Vision-Based Adjustable Projectile Launcher

Vision-based feedback is an ongoing field of research in robotics with many applications. Vision systems are used widely in applications such as self-driving cars, factory lines, and aerial analysis. Self-driving cars that use vision to detect pedestrians, traffic and make decisions based on the vision feedback are being developed (Éloi Zablocki,2022). Quadrotors that use vision to navigate are being researched and tested (Foehn,2022). The goal of our project is to develop the system needed to accomplish Task 7 of the RobotX Competition using vision-based feedback. Task 7 involves identifying the correct dock with the target, navigating to that dock, and launching a 2-inch diameter racquetball through a hole in the target (Robonation, Task 7- Find and Fling, 2022). To this end, a projectile launcher will be developed, capable of launching the four racquetballs provided. The launcher will be interfacing with Team AMORE’s WAM-V, an unmanned surface vessel equipped with navigation and vision systems (Team AMORE, 2022). Code will be developed that allows the WAM-V to identify the required target via the onboard ZED 2i camera, and navigate to that target. As the ball is launched at the target, vision-based data will be collected using the camera, about the position of the ball as it either passes through the goal, or as it misses the target. This data will be used to adjust the onboard simulation controlling the launcher’s firing velocity and release angle, in order to increase its accuracy. Doing so, the more shots are fired, the more accurate the shooter will be.

The Effects Host Have on the Microbiome of Michigan Ticks

The objective of this research project is to see if there is any relationship between the last host of Michigan ticks and the bacteria in their microbiome.

Presence of Taxoplasmosis within Felines and Canines in the Upper Peninsula Animal Shelters

Looking to see if Toxoplasmosis is prevalent in Sault Ste Marie within felines and canines at the animal shelter. Toxoplasmosis is an illness that comes from the Toxoplasma gondii parasite, also known as “Crazy Cat Lady” syndrome. This parasite has the ability to change the behavior of the intermediate hosts to be eaten by the definitive host. Toxoplasma gondii infects the Felidae family, because they produce a bunch of linoleic acid that is necessary for the parasite to fully mature and complete its life cycle. The parasite can enter the brain of a fetus if a pregnant woman is infected, and it can cause adverse effects for immune compromised individuals. Knowing the prevalence in our area will help to better manage risks to this parasite.

Comparing Cognitive Effects of Pharmaceutical Anxiety Medications with Holistic Alternatives

Pharmaceutical anxiolytics, while being decently effective, also come along with a plethora of neurological side effects such as lethargy, sedation, ataxia, and even seizures (Ballantyne and Ciribassi, 2014; Sargisson, 2014). The most notorious among these medications is Trazodone, a selective serotonin reuptake inhibitor (SSRI) which has been known to commonly cause stress on the cardiovascular system (Atli et al., 2019). Because Trazodone is an SSRI, patients that use this medication are also at risk of developing serotonin syndrome, which can be deadly in extreme cases (Boyer and Shannon, 2005). Due to the adverse effects that pharmaceutical anxiolytics can have on animals, many pet owners have been looking toward holistic alternatives to ease their pets’ anxiety, such as melatonin supplements. Adding melatonin supplements to an animal’s diet has been observed to reduce situational anxiety and even inhibit memory impairment (Lamtai et al., 2021).

Considering the fact that pharmaceutical medications have neurological side effects that are observable even to the untrained eye, there could be a possibility that the medication is doing more harm in more subtle ways that owners may not notice. The purpose of this study is to determine if these pharmaceutical anxiolytics do, in fact, hinder the cognitive processes of animals by comparing them to holistic Should these medications prove to alter the cognitive ability of the subject taking them, this could affect the overall treatment for anxiety in domestic pets. Other methods of treatment would need to be sought out to replace pharmaceutical anxiolytics for all animals. In order to test if a pharmaceutical anxiolytic, specifically Trazodone, has any effect on the cognitive processes of an animal, 18 Long-Evans rats will be divided into three groups of six subjects. One group will act as a control, another group will be administered Trazodone, and the last group will be administered melatonin supplements. Before each group is treated with these medications, baseline testing will be performed, wherein each rats’ working memory, spatial memory, and mental health will be assessed. A four-week treatment period will follow, and afterward the rats will be tested using the same methods as the baseline testing. Once the data has been compiled, it will be determined whether or not pharmaceutical anxiolytics negatively affect the cognitive processes of animals.

Plant Tumor Cells’ Effects on Glycolysis

Summary: One of the hallmarks of cancer is an increase in glycolysis in the presence of oxygen (Jiang, 2017). This is known as the Warburg Effect. Though it is not fully understood, it is thought to result from mitochondrial dysfunction of the cancer cell. I would like to take a closer look at this effect using a novel model plant disease, termed crown gall. In this experiment, I will be inoculating soybean plants with Agrobacterium tumefaciens to induce the formation of crown gall tumors and quantifying a difference of the level of pyruvic acid between the uninoculated plants (control group) and the groups of inoculated plants with varying levels of inoculation. Introduction: Both plants and animal cells have mitochondria that work to produce energy for the cell. In the presence of oxygen, plants will undergo cellular respiration to acquire ATP, which is used for a multitude of cellular processes requiring energy. However, when oxygen is not available the cell will undergo glycolysis in the cytoplasm. Glycolysis is a much less efficient way to gain ATP as it only produces 2 ATP whereas cellular respiration occurs in the mitochondria, the powerhouse of the cell, and produces 30-32 ATP. The Warburg Effect, or Warburg’s hypothesis, states that “cancer cells choose aerobic glycolysis as [a] main mode of glucose metabolism instead of more efficient oxidative phosphorylation” (Jiang, 2017). Cancer is the uncontrolled division of cells that form masses of tissue called tumors. Cancer cells also have dysfunctional mitochondria that causes the cell to rely on glycolysis that can be done in the cytoplasm outside of the mitochondria (Fadaka et al, 2017). Agrobacterium tumefaciens is a gram-negative bacteria found in soil that causes crown gall tumors at wound sites of the infected plants (Hooykaas et al, 1982). The bacterium enters the plant through wound sites and transforms the healthy plant cells into tumors (Lee et al, 2009). Soybean plants are a plant that take approximately 45-55 days to mature. They require warm soil to grow and grow best in daytime temperatures around 70°F (21°C). They are poor soil tolerant and require minimal space usage for potting and soil (McConnell, 2019). Pyruvate, or pyruvic acid, is produced by the final step of glycolysis and can be used to measure how much glycolysis is being undergone in a sample. Lactate Dehydrogenase (LDH)-based Spectrophotometric assay is the most common way to test pyruvate concentrations. It is a well-established assay compatible for taking these measurements from my samples. Importance: There could be a correlation between bacteria infecting an organism that causes tumors (cancerous) cells and the level of dysfunction of the mitochondria. This is important because of all the other possible implications and questions that follow afterward.

If there is an increase in glycolysis that correlates with the level of inoculation of the bacteria, further testing could be done to understand the correlation. Is there a correlation between the amount of bacteria inoculum and the amount of glycolysis? How do the glycolysis levels correspond (or don’t) with the actual tumor formation? These are important questions for understanding when the cell’s metabolic “transition” occurs relative to the actual cancer (tumor) formation. Methods: There will be four groups of three plants each so all measurements will be done as a mean of the triplicate. A group of three plants will have no bacteria inoculation and will be used as a control. Other groups, all of three plants each, will be: single-wound inoculation, double-wound inoculation, and triple-wound inoculation. This will bring the total of plants tested to 12 which fits well on a 96-well plate used for spectrophotometry which will be used to measure the absorbance and calculate the pyruvate concentration. The soybean plants will be grown in the university greenhouse to ensure the most controlled environment possible, to increase repeatability and replicability of future experiments. Plants will be grown for 55 days to ensure they have reached maturity (McConnell, 2019). A small incision will be made along the stem of the plant and the wounds will be inoculated with the same relative number of bacteria cells of Agrobacterium tumefaciens, using an inoculating loop. Agrobacterium tumefaciens is a Biosafety Level 1 (BSL-1) organism, so it is compatible for safe use by student researchers. Additional incisions and inoculation will be done to create the varying groups (one wound, two wound, and three wounds). A visible gall tumor can be seen in 4 weeks, so plants will be monitored for 4 weeks to ensure the proper number of crown gall tumors have grown for each inoculated plant (Gonzalez-Mula et al, 2019). The pyruvate extraction procedure will be the modified procedure done by Zhu et al (2010) that was adapted from the original procedures from O’Donell-Tormey et al (1987) and Lamprecht et al (1974). A standard curve of absorbance and pyruvate concentration will be created using known concentrations of pyruvate extract. The absorbance will be taken at 339 nm, LDH will be added, and the sample will be incubated at room temperature in the dark for 30 minutes before absorbance is taken again. The change of absorbance between these two measured absorbances will be used to calculate the pyruvate concentration for the standard curve. Extraction of LDH will be done by the blending of the soybean plants to homogenize them. Cells will be cultured and treated with extraction buffer, centrifuged, and put through a column to purify the LDH (Rivoal et al, 1991). The LDH assay will be repeated on the cells after purification treatment. Absorbance will be taken and used with the standard curve to calculate the pyruvate concentration (Zhu et al, 2010).

Conversations with Dogs: Does Talking to our Canine Companions Reduce Their Stress?

Humans and canines have lived together for thousands of years, but the bonds that once were simply companionship have become reliance on one another. The anatomical structure of a dog’s face and the expressions they use have even been influenced by their relationships with humans by evolving to mimic those of their human counterparts (Smith and Valkenburgh 2020). These have been recognized, sometimes subconsciously, by humans in need when they interact with therapy dogs. The simple presence of canines has been shown to reduce human stress in situations varying from patients in emergency rooms (Kline, Fisher, et al. 2019) to overwhelmed college students (Fiocco and Hunse 2017). However, humans are not the only ones whose stress can be reduced from interactions with the other species.

A study conducted in the United States found that shelter dogs who were fostered overnight had decreased stress levels, based on behavioral characteristics and cortisol:creatinine ratios in the urine, when compared to nights they spent without human contact in the shelter (Gunter, Feuerbacher, et al. 2019). Another study found that petting dogs after they underwent a blood draw reduced the stress levels of the animal during a subsequent blood draw 20 minutes later (Hennessey, Williams, et al. 1998). Interactions such as these show that though an emphasis is placed on the emotional support humans receive from their canine companions, the dogs feel the benefits of the relationship, as well. Many studies have been conducted to discover the impact of our interactions with dogs, but most focus on the direct contact between the species, such as petting. Few to no studies have examined the impact of day-to-day vocalizations with canine companions. The objective of this study is to examine how human voice and conversation impacts the stress levels of dogs. The hypothesis is that human vocalizations will reduce the behavioral signs of stress in dogs as well as lower the urine cortisol:creatinine ratio. Methods A total of ten dogs will participate in the study, all of which will be from eastern Upper Peninsula animal shelters. A veterinarian will be on site while the research is being conducted to ensure the dogs remain healthy and handled correctly. Since staying in a kennel has been found to cause stress in dogs, this will serve as a natural stressor for the experiment (Hennessey, Williams, et al. 1998). After being kenneled for two hours, each dog will spend ten minutes interacting with a human. A baseline behavioral assessment will be recorded during initial interaction with the dog to compare to the final results. Salivation, panting, pacing, and vocalizing will be graded on a scale of 0-5, with zero signifying that the behavior is not present, and five signifying that the behavior is excessive. All of these behaviors have been recognized as visible signs of stress in canines (Almond, 2018). During the ten minutes on the first day, the dogs will be spoken to in a normal voice and petted. The same phrases to greet the dogs using their specific names, discussing the plan for the day, etc. will be used. After the ten minute interaction with vocalization, the dog will be walked outside to void their bladder, and a urine sample will be collected (free-catch using a collection cup). The sample will be labeled with the time of collection, date, name of the dog, and “vocalization,” and will be frozen until data analysis occurs. The same behaviors as before will again be observed and recorded after this procedure to determine if signs of stress decreased after the human interaction.

This process will be repeated for each participant on three separate days. To obtain a control, the human interaction will include no vocalization, and the same procedure will be used to record stress behaviors and collect the urine sample. However, on these days, the dog will not be spoken to at all. Interaction will be limited to human presence and direct contact (petting). This will also occur on three separate days, resulting in data from each dog for six days, three with vocalization, and three without. For urinalysis, the cortisol:creatinine ratio will be measured. Methods for this analysis were used in a study conducted to determine if this ratio is a valid measurement of increased stress in dogs. The study found that by measuring the amount of cortisol using an immunoassay kit and comparing that value to the amount of creatinine in the sample (determined by Jaffe reaction and spectrophotometry), the ratio was a valid indicator of stress in canines. Cortisol is a hormone that is released when canines are stressed. An increase in the cortisol present in urine correlates to an increase of feeling of stress in a dog. The creatinine content is used as a measure of urine concentration, which varies by animal, hydration, and other factors. By comparing cortisol levels to the measure of creatinine, the effects of concentration can be eliminated. Spectrophotometry, the measure of a solution’s light absorption, will be used in this experiment to analyze the urine samples for both cortisol and creatinine (Part, Kiddie, et al. 2014). These values, as well as the values assigned based on stress behaviors will be compared for each individual dog for vocalized interactions versus interactions without vocalization. If the cortisol:creatinine ratio and stress behaviors with vocalization are lower than the values without vocalization, it can be determined that human speech and conversation lowers canine stress.