1. Biodiversity of Long Island Macroinvertebrate In the Connetquot River
Brianna Marinick, Michael Hanson, Samantha Kelly, Isa Hollander1
1Connetquot High School
work hard ladies! (at the beginning)
Abstract
Many macroinvertebrates are vulnerable to varying water quality and climate change, which makes them an ideal specimen to see how marine life is being affected by the changing factors. Macroinvertebrates were collected from the Connetquot River in three locations with the goal to prove whether or not the biodiversity of macroinvertebrate species has decreased in the Connetquot River over the last few years. After the samples were collected, their DNA was extracted and PCR was performed. After completing the PCR process, the samples were put into gel electrophoresis to see whether or not the COI gene was present and was then sent to Barcode LI for sanger sequencing. Once the sequences were sent back, they were processed through DNA subway to create barcodes and phylogenetic trees. After the barcodes and phylogenetic trees were created, barcode sequences from the Bold Systems database were used to compare to the samples that were collected at Connetquot River. After comparing five species to our samples, it was discovered that the species identical to most of our samples was OPC-2018.
Introduction While warmer weather may be beneficial to warm water species, indigenous species that are used to stable weather experience negative effects such as extinction from invasive species and a lack of food or other factors in places such as Long Island.1 Freshwater organisms, also known as scuds, have a higher chance of being affected due to their isolation.2 Fresh waters are particularly vulnerable to climate change because many species within these habitats have limited abilities to disperse as the environment changes.1 Scuds inhabit all bodies of water on Long Island.3 We hypothesize that biodiversity has decreased over the last few years due to temperature changes. Macroinvertebrates in the Connetquot River are good indicators of climate change because they are common in many locations and are susceptible to climate change and pollution. This can help measure the health of the Connetquot River because it can be seen whether or not macroinvertebrates had to go through mutations or died off due to climate change and pollution from the DNA sequences that were produced.
Materials & Methods
The samples were collected at Connetquot River, at three locations; the main pond, the big bridge, and above the hatchery. To collect macroinvertebrates, a net attached to pvc pipe in the shape of a circle with a pole sticking up was used so the scuds can be easily collected. The net was put in the water and with a little bit of dirt so that the net didn’t move. After ten minutes, the net was taken out of the water and any scuds, or anything that looked like a scud, was picked out of the dirt. This step was repeated until we got twenty samples. Once the DNA was extracted with the silica method, PCR amplified using the primers LCO1490 and HC Gel electrophoresis was used to separate the DNA and to help see which samples worked in PCR. It was also used to see which ones had the COI gene. Then DNA subway was used to process the sequences, generate barcodes and compare samples to databases.
Results Twenty-four samples were collected, DNA was then extracted and later put through the PCR process. Gel electrophoresis was done and showed to have been successful on only 12 of the samples. The DNA of the 12 samples was sent to Cold Spring Harbor Laboratory to get sequenced. The data was sent through the Barcode LI database and the results showed that there are 8 samples, PKX-002, PKX-005, PKX-009, PKX-010, PKX-016, PKX-017, PKX- 019, PKX-023, of the 12 samples that got sequenced which match the un-identified species of Proasellus, kno swn as Proasellus sp. OCP Sample PKX-024 was found to match none of the other samples.
PKX-005
PKX-024
Discussion
The results show that there are possible matches, but there is no definite match and therefore we couldn’t get the organisms down to the species level. Based on the pictures that were taken, most samples look to be of a similar species. However, when looking at sample PKX-024, there are differences such as the shape and overall appearance. After performing PCR and putting the data through DNAsubway, it’s clear that PKX-024 is a different species known as the sialis larvae. While this sample is different, the other samples were at least identified to the Proasellus genus. This means no definitive conclusion can be made on the biodiversity of the Connetquot River over time, as we did not obtain enough samples. While data was being collected, there were some issues that we faced, such as a few of the samples having a failed DNA extraction, which would lead to failed barcoding, which would prohibit our ability to identify all the samples. Future experimentation may be improved by being more careful during the DNA extraction process and collecting more samples to hopefully expand the number of different genus’ and species.
References
Woodward G, Perkins DM, Brown LE. Climate change and freshwater ecosystems: impacts across multiple levels of organization. Philosophical transactions of the Royal Society of London. Series B, Biological sciences Jul 12 [accessed 2019 May 15].
Johansson MP, Quintela M, Laurila A. Genetic divergence and isolation by thermal environment in geothermal populations of an aquatic invertebrate. Journal of evolutionary biology Sep [accessed 2019 May 15].
Copyright Global Sea Temperatures - A-Connect Ltd. Long Beach Sea Temperature May Average, United States | Sea Temperatures. World Sea Temperatures. [accessed 2019 May 15].
Acknowledgements
We would like to thank Cold Spring Harbor for providing us with materials, our science research teacher Mr. Halloran, a Day in the LIfe for letting us collect the samples, the BOLDSYSTEM database for helping to identify the samples and last but not least, DNASubway for helping us with our samples’ DNA sequences.