The Biodiversity of Ants in a Northeastern Suburban High School - PowerPoint Presentation

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The Biodiversity of Ants in a Northeastern Suburban High School

Julie Kadletz1, Van Gatz2, Caleigh Cantelupo2, & Dr. Jennifer Gatz31Shoreham_Wading River HS, 2Riverhead HS, 3Patchogue – Medford HS

Results

Abstract

DNA barcoding is an important measure for determining variations in mosquitoes of similar phenotypes. DNA Barcoding will be used in addition to traditional taxonomic methods to scientifically identify mosquitoes collected in three different locations on eastern Long Island. Mosquitoes are a problem on Long Island, and the warm humid climate in summer is conducive for them to increase in population size. Mosquitoes pose a threat to the public as a vector for disease. Recently, eight mosquito samples collected in late July throughout Suffolk County have tested positive for West Nile virus. Another local mosquito sample tested positive for a rare and possibly deadly virus called Eastern Equine Encephalitis. While there is a vaccine for horses against the virus, there is no vaccine for humans (Coburn, 2019).

We used a combination of a sugar and yeast solution, and dried ice in our traps to capture mosquitoes overnight in three different locations, from the north shore of Shoreham and Wading River, as well as the Wildwood Lake area in Riverhead. We hypothesized that the biodiversity of mosquitoes may be greater near the wetland area of Wildwood Lake, and its proximity to Manorville where mosquitoes carrying Equine Encephalitis were identified. The insects were photo-documented and DNA barcoding was used to identify species. By understanding what types of mosquitoes are in the Peconic area, we may be able to identify increased risk that certain species may pose as potential disease carriers.IntroductionThe goal of this study is to determine the species of mosquitoes breeding in the three different locations that we collected samples from. We want to recognize if the different kinds of land, wetland, and the north shore area, would affect mosquitoes' biodiversity. The question of this study is to what extent does the biodiversity of mosquitoes increase the chance of disease spread to humans? We will compare the DNA sequences we obtain from our samples collected and compare them to a base sequence. Once we know what type of species they relate closely to and how many different kinds of species we can identify in each area, we can study the potential that they have for being a vector for disease and notice if the biodiversity differs in each of the different areas. Therefore we might be able to inform the public of areas that may be at risk for getting an infection or illness via mosquito transmission. Materials & Methods We initially used dry ice as an attractant with the mosquito lure and three separate BG Sentinel-2 mosquito traps for the three locations. A second capture trial was needed in location two so, we supplemented a sugar and yeast solution in our trap. This second trial yielded a higher frequency of mosquitoes in location two, which may have presented a disadvantage to the other two location sites. Mosquitoes were trapped over a 24 hours from dusk to dusk the next day. The insects collected from the three locations will be photo-documented, and DNA barcoding will be used to identify species and if they are carriers of any type of disease. The BG Sentinel-2 trap is a piece of professional equipment designed to capture mosquitoes. It is challenging to collect mosquitoes on windy or rainy days, as mosquitoes are less likely to be out and about. The trap will be placed on the ground with dry ice and a chemical lure placed inside. The trap has a lid that has an attached intake funnel. To turn on the fan that will activate the intake funnel, we plug in a large battery. For the fan to start, the lid of the trap must be closed. As soon as the lid is closed and the battery wires are plugged in, the intake funnel will activate. The trap will be left outside for 24 hours. Since no mosquitoes were collected in site two, I replaced the dry ice with a sugar and yeast solution. When the collection is complete, I will unplug the battery and close the intake lid as soon as the battery is unplugged to avoid any mosquitoes from flying out. I will take the entire funnel and place it into a large plastic bag. To ensure the least amount of damage to the mosquitoes, the plastic bag will be placed into the freezer for a minimum of two- hours to allow the mosquitoes to die. After two hours, I will take the netting off of the funnel and dump all of the dead mosquitoes into a container and leave it in the freezer until the DNA barcoding takes place. DNA Extraction We began the DNA extraction process at Brookhaven National Laboratory. We started by ensuring that we used a series of sterile equipment such as tweezers, pipette, containers, etc., to avoid contamination of samples. We selected six samples from each collection site. To extract the DNA from the mosquitoes, we used a silica extraction method. This method is where DNA is separated by nucleic acids that are binding to silica resin. We preserved the mosquito DNA by storing it in 96-100% ethanol. We took two control mosquitoes, samples one and twenty-three, to look back on and make sure DNA was extracted correctly. Gel Electrophoresis To determine which samples we could send to be barcoded, gel electrophoresis was done for small amounts of each sample. We could see which samples contained DNA, and we were able to determine which samples could be sent off to be barcoded. Figure 1 shows if the samples that had DNA in them, the luminescent ones are the ones that were sent off to be documented. From site one, samples 2,5, and 6 were sent off to be barcoded. From site two, samples 17, 18, 26, 27, 29, and 30 were sent off to be barcoded. And from site three, samples 32, 33, 34, 35, 36, 37, and 38 were sent off to be barcoded. We sent the samples that contained DNA to Cold Spring Harbor Lab, where they referenced us to a DNA Subway program, we were sent to the MUSCLE database to determine the type of species of each mosquito. From there, we can determine the amount of biodiversity at each site.

Discussion Figure 1 shows that site one (samples PXR002 and PXR005) collected from the wetland of Riverhead with the coordinates -72.6734, 40.8996, contained the mosquito species punctipennis and albopictus. The sequence that matched to the site one mosquitoes JQ388786|aedes_albopictus and KR666470|anopheles_punctipennis. Sample PXR005 fit with the punctipennis species, and sample PXR002 matched most closely with the albopictus species. Figure 3 shows the comparison of sample PXR005 and the punctipennis mosquito species. Figure 4 shows the comparison of sample PXR002 and the albopictus species. Site two (samples PXR009-PXR030), collected in the north shore of Shoreham with the coordinates -72.9034, 40.9529, showed that most of these samples related novaeangliae and matched with the sequence KJ444714.1|limonia_novaeangliae. Figure 5 shows the comparison of sample PXR026 with the novaeangliae mosquito species. However, sample PXR018 was incorrectly chosen to be barcoded and not a mosquito because of the apparent genetic difference. Site three (samples PXR032-PXR038), collected in the north shore of Wading River with the coordinates -72.811 , 40.9587 seemed to have matched to the antiqua species and matched to the sequence KJ208351.1|delia_antiqua. Figure 6 shows the comparison of the antiqua species to sample PXR036. We see that in the north shore area, we get the same two species of mosquitoes in both sites, the novaeangliae and antigua species. In the wetland of Riverhead, we see a set of two totally different species. By comparing the samples from each different location, we see that each location has different types of species. Site one had two different mosquito types, and sites two and three only had one kind of mosquito. This supports our hypothesis that site one would have had a more considerable amount of biodiversity due to its location and climate. Site one was collected from the wetland area in Riverhead near Wildwood Lake. As stated above, a warm, humid environments are conducive for mosquitoes to increase population size and therefore increase the probability of disease and infection being spread. There is a higher risk of mosquitoes spreading and transmitting disease if there is a more considerable amount of biodiversity. We can use this information to inform the residents that live over there to be more careful with the mosquitoes in the summer and tell them that they are more likely to get an infection with a mosquito-borne disease because of the high population of mosquitoes there due to the climate. AcknowledgementsA big thank you to Dr.Gatz, Dr. Perez at Brookhaven National Lab, and Cold Spring Harbor DNA Learning Center for helping make this project possible.

Figure 1.

MUSCLE sequence alignment viewer retrieved from DNA Subway.