Surveillance of Microplastic Pollution in

Central Virginia Freshwater Ecosystems

Alexandra Reddy
Department of Biology and Chemistry
 Outline
1. Overview of Plastics
2. Characterization of Plastic Pollution
3. Impacts of Plastic Pollution on Aquatic Organisms
4. Marine vs Freshwater Microplastics
5. Bridging the Gap This Project
6. Methodology
7. Results + Discussion
8. Conclusions
9. Future Directions
 The Origin of Plastic
• Synthesized in 1907
• Cheap
• Minimal energy
• 20 major types (North and Halden, 2013)

Retrieved from https://www.recycleandrecoverplastics.org/plastic- Retrieved from Retrieved from https://www.dailydot.com/irl/straw-ban-ableism-

materials/plastic-bottles/plastic-bottles-istock-926200658-medium-2/ http://exclusive.multibriefs.com/content/walmart-joins- restaurant-thread/
worldwide-effort-to-reduce-its-plastic-consumption/waste-
management-environmental
 Plastic Production
• 1950 1.5 million tons
• 2014 299 million tons
• 2019 1.3 billion tons
• 8 million tons enter the oceans (Nelms et al., 2015)

Retrieved from https://www.sundried.com/blogs/news/how-bad-is-the-global-plastic-pollution-problem-and-what-can-we-do-

about-it
Distribution of Plastic Pollution

Retrieved from https://ourworldindata.org/faq-on-plastics

 Categories of Plastic Pollution
• Large plastics
o Entanglement

o Ingestion with visible effects

Retrieved from https://inhabitat.com/peanut-the-story-

• Small plastics
behind-the-poor-sea-turtle-deformed-by-a-six-pack-ring/

o Ingestion

o Gastric obstruction

o Physiological effects

o Trophic transfer
Retrieved from
https://ocean.si.edu/ocean-life/seabirds/laysan-albatrosses-
 What are microplastics?
• Larger pieces quantified into smaller pieces
– Wave movements
– Ultraviolet light
– Physical abrasion

Retrieved from https://www.brushmable.com/blogs/mable-lifestyle/microplastics-the-smallest-big-problem

What is classified as a microplastic?
• Classified according to length
• <5 mm considered to be microplastics

(Oberbeckmann et al., 2015, p. 554)

How do microplastics impact aquatic organisms?
• Respiration

• Direct or indirect ingestion

• Gastric obstruction

• Physiological effects Retrieved from https://www.odt.co.nz/news/national/still-waiting-—-long-road-marine-protection-

area

– Liver toxicity

– Endocrine disruption

– Pesticides

– Microorganisms
Retrieved from https://www.worldwildlife.org/stories/what-do-sea-turtles-eat-
unfortunately-plastic-bags
 Primary vs Secondary Microplastics
• Primary
– Micron size
– Industrial production:
textiles,
pharmaceuticals,
cosmetics
• Secondary
– Fragmentation and
degradation
– Agriculture, clothing,
fishing, automobile, Retrieved from https://encounteredu.com/multimedia/images/sources-of-microplastics

household
How are microplastics transported throughout the food
chain?

(Medrano et al., 2015, p. 76)

 Outline
1. Overview of Plastics
2. Characterization of Plastic Pollution
3. Impacts of Plastic Pollution on Aquatic Organisms
4. Marine vs Freshwater Microplastics
5. Bridging the Gap This Project
6. Methodology
7. Results + Discussion
8. Conclusions
9. Future Directions
 Marine Microplastics
• Well-researched
• Documented in a variety of species
– Sea turtles
– Dolphins
– Rays
– Seals
Retrieved from https://ars.els-cdn.com/content/image/1-s2.0-
S0269749117343294-fx1_lrg.jpg
 Freshwater Microplastics
• Research is scarce
• Particularly for freshwater invertebrates
– Macroinvertebrates
– Amphipod crustacean
– Water flea

Retrieved from https://www.curriculumnacional.cl/614/w3-article-25330.html

 Bridging the Gap
• Keystone species Crayfish
• Contribute to freshwater
microplastic knowledge
• Surveillance of local
microplastic pollution

Retrieved from https://bugguide.net/node/view/1157618/bgpage

 Outline
1. Overview of Plastics
2. Characterization of Plastic Pollution
3. Impacts of Plastic Pollution on Aquatic Organisms
4. Marine vs Freshwater Microplastics
5. Bridging the Gap This Project
6. Methodology
7. Results + Discussion
8. Conclusions
9. Future Directions
 Sample Collection
• Two stream locations:
1. Opossum Creek
2. Rock Castle Creek
 Sample Measurements and Dissection
• Total length (mm)
• Blotted wet mass (g)
• Gender
• Dissection
– Dissecting microscope
– Scanning Electron Microscope
Retrieved from https://microscopecentral.com/products/asbestos-stereo-microscope

– Glass jars
 Sample Preparation for Chemical Analysis

• Ultrasonication
• Membrane filtration
– 25 mm stainless steel Retrieved from Retrieved from https://laboratory.pall.com/en/sterile-

– Glass microsyringe
https://www.thomassci.com/scientific- filtration-clarification/filter-holders-membranes/
supplies/Filter-Holder products.html

– Glass jars

Retrieved from https://www.grainger.com/product/AIR-TITE-Reusable-Glass-Syringe-

19G355
 FTIR-ATR
• Chemical fingerprints Functional groups
• Most reliable technique

Retrieved from https://www.slideshare.net/savvysahana/atr-ftir-sahana (Ory et al., 2018, Supplementary Material Handout p. 6)

Classification

(Cozar et al., 2017, Supplementary Material Handout Figure 4)

 Outline
1. Overview of Plastics
2. Characterization of Plastic Pollution
3. Impacts of Plastic Pollution on Aquatic Organisms
4. Marine vs Freshwater Microplastics
5. Bridging the Gap This Project
6. Methodology
7. Results + Discussion
8. Conclusions
9. Future Directions
 Preliminary Investigation
• Carolina Biological Supply
Company crayfish
• Two hard particles
– Speculated to be foreign
contaminants
 Preliminary Investigation
• Polyethylene filled with calcium carbonate
– Filler for white lawn furniture
• Components of the gastroliths
• Identities inconclusive, but compelling
Rock Castle Creek Findings
Fiber vs Commercial Translucent PP
102

100

98

96

%T (scaled)
94

92

90

88
%T RCfiber1 201419 32s
86
%T PP translucent; scale and shift
84
3500 3000 2500 2000 1500 1000 500
wavenumber (cm -1 )
 Tank Study
• Inadvertent vs intentional ingestion
• Tank study Feeding behavior
• 8 crayfish from Opossum Creek
– 8 individual glass aquaria
– Randomized to a plastic treatment:
plastic bag, synthetic fabric, or control
 Tank Study
• Dissected after 5 months
• No microplastic particulates
• Shredding behavior
• Interpretation:
1. Did not target and ingest
2. Did target, ingest, and excrete
 Gill Filament Analysis
• Filter water over gills
• Entrapment of plastic
fibers
• Blue fiber identified in
an Opossum Creek
crayfish
 Gill Filament Analysis
• 10 Rock Castle Creek crayfish
• 4 fibers identified
 Outline
1. Overview of Plastics
2. Characterization of Plastic Pollution
3. Impacts of Plastic Pollution on Aquatic Organisms
4. Marine vs Freshwater Microplastics
5. Bridging the Gap This Project
6. Methodology
7. Results + Discussion
8. Conclusions
9. Future Directions
 Big Picture
This investigation strengthens the assertion
that plastic pollution pervades our aquatic
ecosystems and has negative implications for
aquatic life.
 Bridging the Gap
• Preliminary investigation Baseline
• Limited knowledge of freshwater pollution
– Particularly freshwater invertebrates
• Support investigation of microplastics in
crayfish
 Future Directions
• Investigation of species in higher trophic levels
– Biomagnification of microplastic particulates
• Investigation of the microbiota of microplastic
particulates
– Influence on the normal microbiota of crayfish
Questions?
 References
• Cozar, A., Marti, E., Carlos, M., Garcia, J., Sebille, E., Ballatore, T., . . .Irigoien, X. (2017). The Arctic Ocean
as a dead end for floating plastics in the North Atlantic branch of the thermohaline circulation.
Science Advances, 3,1-8.
• Medrano, D., Thompson, R., & Aldridge, D. (2015). Microplastics in freshwater systems: A review of
the emerging threats, identification of knowledge gaps and prioritization of research needs. Water
Research, 75, 63-82.
• Nelms, S.E., Duncan, E.M., Broderick, A.C., Galloway, T.S., Godfrey, M.H., Hamann, M., Lindeque, P.K., &
Godley, B.J. (2016). Plastic and marine turtles: a review and call for Research. ICES Journal of Marine
Science, 73(2), 165–181.
• North, E., & Halden, R. (2013). Plastics and environmental health: the road ahead. Reviews on
Environmental Health, 28(1), 1-8.
• Oberbeckmann, S., Loder, M. & Labrenz, M. (2015). Marine microplastic-associated biofilms – A
review. Environmental Chemistry, (12), 551-562.
• Ory, N., Chagnon, C., Felix, F., Fernandez, C., Ferreira, J., Gallardo, C., . . .Thiel, M. (2018). Low
prevalence of microplastic contamination in planktivorous fish species from the Southeast Pacific
Ocean. Marine Pollution Bulletin, 127, 211-216.
• All figures not noted by a “Retrieved from” with a URL link are personal figures that I created.