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Monsignor McClancy Memorial High SchoolMentor: Ann T. SmithTeam Members: Gabriella Sikorski, Randy Singh, Shannon Connors, Peter Maxham, Lily Para, Megan , Pacione, Elvedina Velovic, Quentin? Holmes, Delme Fernandes, Daniel Martini, Cindy Navarro, Daniel ModzelewskiOUR ORIGINAL ESSENTIAL RESEARCH QUESTION:Does a difference in the phosphorus content of soil in a North Eastern United States Urban Forested Wetland, affect the genetic biodiversity of soil macroinvertebrates (sub-phylum Arthropoda, Insecta Chilopoda and Arachnida)?Possible sources of phosphorus for further analysis: NYC Sewage wastewater overflow due to excessive rain, Invasive species Phragmites as a phosphorus source and sink, close proximity to a golf course using high phosphorus fertilizers. ABSTRACT / INTRODUCTION:Biodiversity is a good measure of how healthy an environment is. Using macro invertebrates as our focus point we can determine if the land that we caught them on is healthy. We performed chemical soil testing and soil texture analysis tests on the soil we analyzed at both locations, in addition to physical items found, such as discarded plastic bottles, papers and mall pieces of old furniture that was buried. Macroinvertebrates are excellent creatures to use to test the health of an environment because each individual species thrives in a very specific environment, making the study of macroinvertebrate biodiversity a clear indicator of whether or not an area is clean. We recently did a simulation in lab called “Benthic Bugs” where we learned about the “Simpsons Diversity Index,” which tells us how healthy a body of water is by the biodiversity of its’ macroinvertebrates. We believe soil the biodiversity of soil macroinvertebrates should be able to indicate the health of soil too. According to the Simpson’s diversity index that we worked on in lab called, "Benthic Bugs and Bioassessment,” benthic macro-invertebrates they can be an indicator of water pollution. We are working to find out if there is a similar tolerance level in soil macro-invertebrates, as well. This information, along with the data from the DNA testing confirming the identity all species will be an excellent indicator as to whether or not this land is healthy enough to support biodiversity of soil macroinvertebrates from the sub-phylum Arthropoda, Insecta Chilopoda and Arachnida (excluding Branchiopoda/crustaceans)Our project is relevant to where we live because our data collection took place less than 15 minutes from one of the Brothers of the Sacred Heart High Schools. It compares the biodiversity of these two locations, which are in close proximity to each other on the fringes of one of the most highly urban areas in the world, New York City, within 55 miles of each other, one on each side of Manhattan. For Msgr. McClancy MHS, our research area at APEC in Little Neck, NY was within 500 feet of Northern Boulevard and within a half mile from the Cross Island Parkway. This area was slightly more urban. This area had to litter visible, but was also kept by a caretaker. There is a sewage overflow adjacent to the property. There have been massive changes to the biodiversity in this area in the past century. According to their website, Alley Pond Park was one a healthy and thriving commercial shellfishery thrived in Little Neck Bay during the mid-1800s, but was forced to close in the early 1900s due to watershed pollution from urbanization of the area. The acres of salt marsh where APEC is now located became dumping grounds for debris from the construction of the Cross Island Parkway in the 1930s and the Long Island Expressway in the 1950s. Into the early 1970s, this neglected area remained in horrific condition, subjected to a constant bombardment of dumping. The landscape was still in terrible shape up until 1979 when APEC was designated a National Environmental Study Area. (APEC)Yochanan D. John, a Queens resident who has extensively explored the creek recounted, "has undergone a remarkable recovery since its nadir in the 1970s, when it was a neglected dumping ground suffering under a legacy of industry and pollution... There were factories everywhere, and raw sewage stinking to high heaven." This area is bouncing back from having been a veritable dumping ground, but is it still affecting the area today, and can soil macroinvertebrates be used as an indicator. Kensinger added, " remediation efforts are still ongoing," indicating that the area is not yet what it once was. (Kensinger) In 1976 this dumping ground became part of the “Wetlands Reclamation Project”, restoring its natural beauty.For St. Joseph’s RHS, on Dismal Pond, which is the South Eastern corner of the Dismal Swamp region of Metuchen, NJ there were no major roads within 500 feet of our research area, and over 100 years ago this area was part of a private farm. The environment from where we collected specimen in New Jersey was more suburban. We saw more physical litter (i.e. plastic bottles) in this location, but the only caretakers on this site have been the students from St. Joseph’s Metuchen science classes and Troop 417. There is a rich history of pollution and changes in biodiversity in the Dismal Swamp region of New Jersey. Because so much of the history is not documented, we conducted several interviews with the Brother of the Sacred Heart, who have owned the land for over a century.According to Brother Lucien Blaine, the area was used as a dumping ground for early farms and a quarry, around the turn of the century Brother Robert Ziobro, BSH, added by the 1940’s the area became more built-up and much of the water was diverted, changing the biodiversity of the area. We even found artifacts of the irons of an old fashioned fireplace that we uncovered as we worked in the soil collecting field samples. There were snakes, rabbits and fox, raccoons and possum falcons hawks and deer, some of which still remain in the area. He noted that the mosquito population has increased and now the Brothers wear head-netting when cutting grass, and there has been a decrease in flies during the summer as compared to 50 years ago. The “Dismal Swamp Preservation Act” of 1974was passed ensuring a restoration of its natural beauty. Both locations were inhabited by Native Americans of the Lenape tribe, and share a rich and beautiful history, but more recently both sites share the fact that they were once used as dumping grounds. About 100 years ago in our own US history, wetlands were not viewed as beautiful places to preserve, but a nuisance that bred disease and mosquitos. Both Dismal Swamp and Alley Pond are within a mile of a major interstate highway, both locations have buildings on the grounds, and both locations were bordered by large main streets with residential homes and other small businesses. In general, the New Jersey site was slightly more suburban in appearance than New York, and we initially believed the area to be less polluted as a whole. Thus far, this project has helped us realize how diverse and different our country can be. Our urban environments offer little forest land, and it is therefore essential that we protect these remaining locations.The use of barcoding could possibly help identifying threats to human health, if we can identify whether any rapid evolution has occurred amongst the macroinvertebrates due to pollution. We have recently seen rapid evolution occurring in the Atlantic killifish. (Klein) In our research we came across a study by the Barcode of Life Initiative of Canada, that we agree with that says barcoding will alleviate the human error factor. DNA barcoding can eliminate mistakes we would have made if we only had a dichotomous key to work with. This gives us a positive and undeniable answer that will use over and over again as we continue to build upon our project that we started this year. An article entitled, “How DNA Barcoding Can Help Science,” helped us to understand that we will be contributing to a global database of DNA barcodes. It may lead to new ideas for preserving the biodiversity by identifying the same organisms in different habitats, we can learn from each other to help conservation. SPECIFIC AIMS:Evaluate chemical differences in soil chemistry and correlate to biodiversity two different environments. To document the biodiversity of two locations that used to be dumping grounds but have gone through changes over the past century, and restorations over the past 40 years, for approximately the same amount of time.To have this become a multi-year project, due to the extensive history we have for both pieces of property from research and living history interviews.To compare the biodiversity between these two locations based on their proximity (or lack thereof) to potential pollutants. Identify which species are more tolerant to environmental differences To determine if similar environments should support similar species in a specific geographic region.Study the burrowing habits of macroinvertebratesThe effect of weather on field sample collectionMETHODS:Seniors from Monsignor McClancy MHS traveled to both St. Joseph's High School in New Jersey, and the Alley Pond Environmental Center in New York.Using gloves, field vials, recovery bins, flags, quadrats and small used tools to gently dig around and scoop them up, we extracted various species of macroinvertebrates from an urban forested wetland, which was abundant in trees and near a body of water. We first flagged the area, laid down our quadrats and dug in the soil and investigating rocks and logs and inside of our field-work areas. We sift through soil on the ground directly if they were large enough, or scooped up the soil with a small sieve and then sifted through it in the recovery bin. Mostly we were able to trap the macroinvertebrate with the tube or with our hands. It was difficult catching some of them as they crawled so quickly. ?Next it was placed into a separate recovery tray that would then make it easier to transfer the macroinvertebrate into a field sampling vial tubeThe field specimens were collected by using sieves and our hands to gently sift through the soil under leaf litter or the fallen trees/logs. Each group member helped in making sure to collect interesting and diverse samples, not duplicated.Next, another group member gently place the living specimens into separate field collection tubes Approximately 10 macroinvertebrate specimens from each regional location were collected out in the field.These macroinvertebrate specimens were then brought back to the lab, photographed and frozen for inspection and DNA processingStudents put each specimen on a piece of 1cm3 graph paper with rulers to take multiple pictures and measurements as well.This process occurred twice, once for each site.We used the LaMotte soil testing kit to perform various chemical tests, such as: wide range pH, nitrogen, phosphorus and potassium. We further completed soil percolation/infiltration testing, mechanical soil analysis using the “Jar Test” and the textural soil triangle.According to the “posts and piling” sampling methods for this year, as indicated in the 2016-2017 BLI welcome email, we conducted our field sampling as follows: In Dismal Pond, NJ, we used the string and stakes method as our sampling technique. First we flagged off the 50x50 target area that we wanted to collect from. Then we collected from two separate quadrats from the ground, inside of our flagged target area: Quadrat 1 - 40.551597, -74.380395 (samples #1-5); Quadrat 2 - 40.551604, -74.380594 (samples #6-10). In Alley Pond, NY we sampled in three separate quadrats, but only used the samples from the first two: Quadrat 1 - 40.762551, -73.752468 (samples #1-6); Quadrat 2 - 40.762338, -73.751975 (samples #7-10); Quadrat 3 - 40.762602, -73.753203 (did not use). This giving us the total of 20 field samples.We have an understanding of the wet-lab work that is to come because we have been studying the videos and downloads available on the CSH website in class. Further, as a freshman we participated in a lab at Cold Spring Harbor to extract DNA from a sample to identify a certain gene. We took a sample of our own cells and extracted the DNA from them. Using a gene amplification device, we were able to make copies and load our own gel electrophoresis tray with our own DNA. From there, we were able to see if we were homozygous dominant, homozygous recessive or heterozygous. I remember that I was one of 2 students to be heterozygous and one of 6 to have the gene at all. This testing allowed us to see if we carried the ALU sequence, a gene that originated in Africa many years ago. The testing allowed us to apply our scientific knowledge and findings to our own lives by helping us to identify things about ourselves that we otherwise never would have known. We understand there will be more work involved in this project, but we understand what is expected of us in our own lab at school.FINDINGS:Upon first observations, our group members believe the specimens found in New Jersey are more diverse than those of New York. From this, we have already discovered that the more harmful chemicals found in pollutants are already present in the NY location, as the phosphorus is already present in the soil samples. We hypothesize that our project could indicate a major difference between the Little Neck, NY and Metuchen, NJ locations if rapid evolution has occurred. We believe the NY area is more polluted on a whole, because of its’ proximity to industrial activities, and we will find less biodiversity in the New York location. We hypothesize the specimens found in NJ will be part of a far more biodiverse community, and thus the biodiversity will not be as affected by pollutants as New York. We have not yet examined the DNA of the specimen only the physical features. Further, we collectively agree that local weather differences may have an effect on the biodiversity of our sampling. All other determinations are awaiting the DNA testing results.On-site chemical testing: LaMotte soil test kit & jar soil test with textural triangle are as follows:Dismal swamp, NJAlley pond env. center, NYResults:wide range pH - 6.0nitrogen – tracephosphorus - tracepotassium - traceSoil type: Sandy Loamwide range pH - 6.0nitrogen - tracephosphorus – highpotassium - traceSoil type: Sandy LoamSAME SAME **FLAGSAME SAME ALLEY POND ENV. CENTER, NY2.761.4%sand4.336.4%silt4.42.3%claysandy loamS.W. DISMAL SWAMP, NJ1.661.5%sand2.430.8%silt2.67.7%claysandy loamSUMMARY:This research project can help us analyze how different environments can affect similar organisms. Although it seems relatively simple, doing the DNA testing by ourselves may be difficult. If we do this correctly we will surely get the definitive answers we were looking for.As a student researcher, there has been a lot that I have learned from being in an AP Environment class, especially in the one I am in. I have been able to physically experience the environment rather than being taught of people who have done so. By actually performing scientific research, my interest in the scientific field has expanded and made me become more aware of certain issues. It brings in a different point of view that I would not have had without this.TEAM INFORMATION:Gabriella Sikorski: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370Gabriella is a senior at Monsignor McClancy High School. She is interested in the Barcode work and has begun considering biotechnology as a possible career choice because of our project.Randy Singh: Randy Singh - I currently attend Monsignor McClancy High School. My intended major in college is a Physician Assistant, I also plan to fulfill my studies at St. John’s University.Shannon Connors: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370A senior at Msgr. McClancy MHS and a member of our school’s cheerleading squad, drama club, National Honor Society, and Campus Ministry. Outside of school she is a dancer and volunteers with Brownie Girl Scout Troop 4415. Peter Maxham: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370I am a senior in Monsignor McClancy Memorial High school. I am an honors student who has achieved honor roll throughout his four years at school. This year he is proud to be a member of the AP Environmental Science class that is working on this project.Lily Para: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370Lily is a senior who enjoys working on citizen science projects, and has enjoyed working on the Billion Oyster Project this year. She likes working as a student-researcher and is now considering careers where research may be required.Megan Pacione: She is currently a senior at Msgr. McClancy Memorial High School, and she hopes to attend John Jay next fall, where she wants to major in forensic psychology while I am there.Elvedina Velovic: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370She is currently a senior and in the National Honor Society, Spanish National Honor Society and is the Captain of the Varsity Softball team. She is planning to go to St. Francis College to study nursing. Quentin? Holmes: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370Quentin is in his senior year and has been an honor and AP student for his entire career. He has already committed to Mississippi State University for baseball. As a student-athlete, he won a gold medal playing baseball in Mexico this past summer for the Pan American games with Team USA. Delme Fernandes: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370Delme is a senior, President of the Student Council and one of the leads in this year’s school play. Delme has a wonderful singing voice and has been performing on stage since elementary school.Daniel Martini: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370Daniel is a senior who enjoys working out doors and with hands-on activities. He especially liked working on this Barcode Long island project because it involved outdoor fieldwork, and that is what he enjoys the most.Cindy Navarro: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370Cindy is a senior who is a hard worker and likes to learn new things. She likes working with power tools and built house wren nesting boxes. She helped her class win the Cornell University contest for case of binoculars with our project, “Full Steam ahead with Stem.” Daniel Modzelewski: Grade 12, Msgr McClancy MHS 71-06 31st Avenue East Elmhurst, NY 11370Daniel is a senior who is on the first-string of the varsity basketball team. He enjoys fieldwork, and he likes working on citizen science projects.REFERENCES:Klein, J. (12/09/16) Rapid Evolution Saved This Fish From Pollution, Study Says. NY Times. . John’s University. $1.3 million awarded for community-based projects to improve health of long island sound. (2014, Nov 19).?Targeted News Service.?Online.Alley Pond Environmental Center , N. (June 9, 2016) In a Queens Park, Reclaiming an Urban Wetland: The once-polluted Alley Creek is slowly but surely making a comeback. Curbed New York. than $1.6 million awarded for community-based projects to improve health of long island sound; more than $900,000 granted to support projects in new york (NY).?(2012). (). Lanham: Federal Information & News Dispatch, Inc. Online. New York State Department of Environmental Conservation. Healthy Water, Healthy People Water Quality Educators Guide. "Benthic Bugs and Bioassessment" by Project WET International Foundation, 2003. Daily. How DNA Barcoding Can Help Science (September 17, 2007) Source: Consortium for the Barcode of Life. , M.A., Fisher, B.L., Hebert, D.N. DNA barcoding for effective biodiversity assessment of a hyperdiverse arthropod group: the ants of Madagascar. Barcode of Life Initiative, Biodiversity Institute of Ontario, Department of Integrative Biology, University of Guelph, Guelph, ON, Canada N1G 2W1 2 Department of Entomology, California Academy of Sciences, 875 Howard Street, San Francisco, CA 94103, USAZiobro, Br. R., Blaine, Br. L. (December 2016) Interview by Peter Maxham ................
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