Undergraduate Research
Committee

A Study of Nesting Preference and Effectiveness of Natural Reeds vs Reusable Wood Trays in Managed Spring Mason Bees, Osmia spp

With the ever-growing awareness of the decline in pollinator species in the United States, which is primarily due to habitat loss but also partially linked to pesticide use and disease spread by the European Honey Bee, a grassroots movement to protect native bees has arisen in the form of providing nesting sites for native pollinators. These man-made nesting sites are commonly known as “bee hotels,” or artificial domiciles, and they are designed to encourage nesting and pollination by native pollinators. However, as the trend has become more popular and the need more urgent, debates have arisen within the native bee raising community over the best way to go about rearing native bees. On the forefront of the debate is which of the two popular commercial nesting materials, natural reeds or reusable wood trays, are more effective in propagating new bees. While many observations – and opinions – can be found on online bee forums, few scientific studies have been conducted addressing the question, with none focusing specifically on spring mason bees, Osmia spp., the most popular native solitary bee reared in the United States for fruit tree pollination.

The gap in the literature is wide, and findings from this study could be consequential to conservation and agriculture alike as the “Save the Bee” campaign broadens to include more than just honey bees. My senior research idea examines two hypotheses: Natural reeds are more effective than reusable wood trays in raising spring mason bees if “effective” is defined as “producing the highest number of viable mason bee cocoons.” Given the option, spring mason bees will show a preference toward natural reeds over reusable wood trays; they will produce significantly more cocoons in reeds than trays when both options are presented to the same bees equally. To compare the effectiveness of different nesting materials commonly used by native beekeepers (namely reeds and trays) I will observe installed spring mason bees, Osmia spp.,and compare the number of viable bee cocoons collected from each nesting type at the end of an active season. On each of the two locations, three groups will be established: Group A: Only Reeds Group B: Only Trays Group C: Reeds and Trays On Site 1, Spring Mason Bee colonies will be established on a property west of Gregory Gardens at 3290 E 9 Mile Road in Sault Ste. Marie, MI. Groups will be placed no less than 200 yards from one another to prevent interaction between colonies. In the spring and summer, the property will have adequate food sources for the installed bees, including crab apple trees, clover, and miscellaneous wildflowers. There is also a creek for a water/mud source. The location for Site 2 is located at 3588 S Riverside Dr, Sault Ste. Marie, MI near the Saint Mary’s River. On both sites, additional “mason bee mud” will also be provided near the domiciles, as mason bees rely on clayey mud to construct brood cells. InvataBee Mason Bee Attractant will also be sprayed on each domicile at the period of installation and when nesting materials are added; InvataBee Mason Bee Attractant is a USDA-patented formula designed to mimic natural pheromones to encourage nesting.

In addition, bird guards will be placed on all domiciles to prevent predation of mason bee larvae. In the fall, cocoons will be harvested, counted, and sorted by site, colony, and nesting material type collected from, for a total of eight bee bags labeled 1A, 1B, 1CR, 1CT, 2A, 2B, 2CR, and 2CT, with the initial number indicating whether cocoons were collected from Site 1 or Site 2 and the following letters indicating colony and nesting material collected from. After the cocoons have been counted by me on-site, they will be sent by mail to Crown Bees Headquarters in Woodinville, WA, to be recounted, tested for viability, and ultimately redistributed to native beekeepers throughout the nation in the following season. Crown Bees will send me a professional breakdown of the number of viable bee cocoons found in each bag, which may differ from the total number of cocoons found in each colony and nesting material due to disease, harvesting errors, and other factors. Bee Bag Grouping Key 1A: Cocoons harvested from the Site 1 Only Reed group 1B: Cocoons harvested from the Site 1 Only Tray Group 1CR: Cocoons harvested from the Site 1 Reeds and Trays group, taken from the Reeds 1CT: Cocoons harvested from the Site 1 Reeds and Trays group, taken from the trays 2A: Cocoons harvested from the Site 2 Only Reed group 2B: Cocoons harvested from the Site 2 Only Tray Group 2CR: Cocoons harvested from the Site 2 Reeds and Trays group, taken from the Reeds 2CT: Cocoons harvested from the Site 2 Reeds and Trays group, taken from the trays A statistical analysis test will be used to determine whether the difference in viable bee cocoon numbers is significant (in other words, if variability in cocoon count is due to chance or nesting material type). If the difference is found to be significant, it may indicate a more effective nesting material, “effective” indicating the successful propagation of mason bees as shown by the higher number of bee cocoons. The secondary hypothesis of this study examines bee preference for nesting material. To determine bee preference, special attention will be paid to the C groups, which contain both reed and tray nesting options, during weekly observations. I will note and document bee activity, indicating which nesting material bees are investigating the most and which nesting material appears to be filled the quickest, particularly at the beginning of the season. In addition, statistical analysis tests will compare cocoon counts in 1CR vs. 1CT, 2CR vs. 2CT, 1CR vs. 2CR, and 1CT vs. 2CT to examine whether preference can be deciphered from cocoon count. Because preference does not necessarily equate to effectiveness level, only the number of cocoons will be considered in statistical tests regarding preference. Viability of cocoons will not be considered.

Relations Between Season and Discharge from a Hydroelectric Reservoir on the Density and Transport Distance of Zooplankton in the Saint Joseph River in Berrien County

Rivers, by their very nature, are a constantly changing ecosystem. River processes can be further complicated if a reservoir is built on the river, with variable discharge entering the river system downstream. For example, water flowing through a reservoir, and downstream from the dam, can be variable by season (e.g., wetter versus dryer months) and storm events, which could increase the outflow of water to a river from a dam. These dynamics can subsequently impact the ecosystem and alter the composition and abundance of certain organisms in a river network. One such important organism group that can be affected by a dam and variable water flow are zooplankton. Zooplankton are part of the food chain of aquatic ecosystems they provide an important source of food for other larger organisms such as fish and can affect water quality. Zooplankton communities can be drastically affected in rivers from seasonal dynamics mitigated through upstream dams and reservoirs (Havel et al. 2009). Although zooplankton are commonly studied in slower moving lakes, zooplankton can also inhabit river systems and be transported downstream to other aquatic ecosystems (Portinho et al. 2016). Some zooplankton species also prefer to inhabit different depths of rivers and can change their distribution day versus night. However, zooplankton remain understudied in rivers versus lake ecosystems, and more information is needed to better understand how seasonal and other environmental factors (such as storms, which are increasing in frequency and intensity in many regions because of climate change) affect zooplankton abundance and transport downstream from reservoirs. My project aims to study the effects of hydroelectric dams on the density and transport of zooplankton in the Saint Joe River in Berrien County.

The effects of storms are also an area of interest for the study as they can increase water discharge, thereby affecting the quantity and composition of zooplankton downstream from what is present in the reservoir. This river also enters into Lake Michigan, and so the amount of zooplankton entering Lake Michigan from an upstream reservoir can be quantified. Samples will be collected from the Saint Joseph River using a 60 µm mesh size, 50 cm diameter, zooplankton tow net. Zooplankton will be collected by a horizontal tow in three locations in the river at each sampling event, the tow net will be administered horizontally from a kayak for 5 minutes to allow enough volume of water to enter to capture enough zooplankton to quantify. These three river locations will be: just downstream of the reservoir, approximately equal distance between the reservoir and the mouth of Lake Michigan, and close to the mouth of Lake Michigan. In the reservoir (Lake Chapin), a vertical net tow will collect zooplankton in a deep section of the reservoir; these vertical tows will collect zooplankton from 1 m from the bottom to the surface. Samples will be collected once a month from May through August and before and after storm events (3 paired before and after storm events will targeted to be collected). After the samples are collected, the zooplankton will be preserved using 99% isopropyl alcohol, and will be analyzed using a dissecting microscope in the LSSU labs to identify each specimen to the genus level. Specimens will also be measured, and density and biomass will be calculated for each sample (Bum et al. 1996).

Effect of Poly-D-Lysine Coating on mRNA Transfection Efficiency in HEK293T Cell Cultures

The ability to make a cell produce a foreign protein (from another organism) is an important tool of cell and molecular biology research with many applications in medicine. In a general sense, some form of nucleic acid, DNA or RNA, needs to enter the cell to provide the genetic code for the foreign protein to be produced. This strategy for protein production in a test tube is the basis for protein subunit vaccines and recombinant protein drugs such as insulin. There are various ways to cause a cell to produce a foreign protein. When not using a virus, this process is known as transfection. One way to accomplish this is with mRNA and various delivery systems. Just as the Pfizer/Moderna mRNA vaccines cause your human cells to produce the coronavirus spike protein, this technology can be used to cause cultures of living cells to produce any protein for which the genetic sequence is known (with limitations.) Compared to cultures of bacteria or yeast, mammalian cell cultures require very specific conditions to survive. This includes: a specific liquid nutrient medium, temperature, gas mixture, and many require a surface to grow upon. The most common material that tissue culture vessels are made of is polystyrene, a plastic. Naturally water repelling, the surface of polystyrene tissue culture plates are treated with oxygen plasma to modify the surface. This results in a negatively charged more water loving surface that allows many cell types to adhere. Some cells require, and others may benefit from, the addition of an extracellular-matrix mimicking coating. This is because cells in mammals are usually surrounded by a matrix of fibers, like collagen. Coatings available include: the polymer poly-lysine and the various animal proteins such as collagen, laminin, and fibronectin. Developed to more closely mimic the extracellular environment of a living organism, the coatings increase the ability of some cell types to adhere to the vessel (Corning, product manual). The aim of this experiment is to assess the effect that a poly-D-lysine coating has on the efficiency of mRNA transfection in HEK293T (human embryonic kidney) cells. In 1977 this cell line was produced from human embryonic kidney tissue, and has been a common cell line for production of recombinant proteins and human viruses. The hypothesis is that the difference the coating presents to the extracellular environment could cause a difference in the development of the cellular membrane. The cellular membrane is the barrier that must be overcome in order for the mRNA to enter the cell and be translated to make a protein.

Therefore, differences in membrane composition or structure could cause differences in the ability of mRNA to enter the cell and be translated into protein. The percentage of cells that the mRNA successfully enters and produces the protein is known as transfection efficiency. The foreign protein being chosen is green fluorescent protein. Under UV light, green fluorescent protein emits a green light. This unique property of fluorescent proteins allows for easy identification, because human cells do not produce any type of fluorescent proteins. Tissue culture plates and tissue culture plates coated with poly-D-lysine will be seeded with the same stock of HEK293T cells. The cells will be grown and handled under the exact same conditions and will differ only in the addition of a poly-d-lysine coating. While growing, the cells will be observed via microscope to observe any difference in the appearance of the cell membrane or other growth characteristics. Once the cells have grown enough they will be treated with mRNA delivered by the common transfection reagent lipofectamine. The transfection efficiency will be determined using the Thermo Fisher Countess automated cell counter. It has the ability to count the total number of cells and the number of cells expressing the green fluorescent protein. Given as a percentage this represents the transfection efficiency. Mean transfection efficiency of coated wells will be compared to the non-coated wells via an appropriate statistical test.

Suicidal Behaviors, ACEs, and Educational Attainment:  A Potential Basis for Developing  More Effective Assessment Tools for Education Professionals

The brain undergoes rapid development during childhood. Experience, both positive and negative, plays an important role in this process. When children experience abuse and/or neglect, it can interfere with their ability to be successful in school. This puts them at risk of not reaching their full potential academically, limiting occupational choices, and ultimately, earning potential. Research has also shown correlations with numerous harmful effects in adulthood (e.g., physical and mental health problems; CDC, 2019) as well as increased risk of suicide and substance abuse (Park, et al., 2021; Whitesell, et al., 2009). Again, the impact to quality of life can be devastating when these children reach adulthood. Therefore, early identification and intervention is crucial. Currently, K-12 schools use the Adverse Childhood Experiences (ACEs) questionnaire as a screener to identify students at risk for poor academic achievement (Catterfeld, 2019; NEA; Pataky, et al., 2019). However, this is only one area of concern related to child maltreatment. Childrens’ suicidal behaviors and initial use of substances often begin before reaching adulthood (CDC, 2019; HHS; 2019). This raises the question of whether the use of the ACEs should be expanded to screen for risk of suicide and substance use to provide a more comprehensive intervention plan. However, a few issues need to be explored. First, the relationship between ACEs and suicide have been investigated with an emphasis on suicide attempts and completions but not suicidal behaviors. Second, a similar research focus exists between educational attainment and suicidal behaviors (i.e., educational attainment has been linked to suicide attempts/completions but not behaviors). Therefore, the purpose of this research is to establish a link between suicidal behaviors, ACEs, and educational attainment that would permit earlier identification of risk before a suicide attempt or completion occur and/or educational attainment is threatened. To explore this question, a sample of 150 adult participants recruited through Prolific will be assessed for ACEs, suicidal behaviors, educational attainment, and substance misuse. Childhood maltreatment will be measured using the ACEs, suicidal behaviors will be examined with the Suicidal Behaviors Questionnaire – Revised (SBQ-R), a question will ask participants what their highest level of education, and the NIDA-Modified ASSIST (NM ASSIST) will be used to characterize substance misuse. If a correlation is found between these variables, it would lend support to expanding the ACEs screener to include suicidal behaviors, giving educational professionals better tools with which to help students succeed.