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Poison Produce?

Proposed Research in Accumulation of Microcystin in Food Plants
Anne C Ewert
Advisor: James F Haney
Department of Biological Sciences, University of New Hampshire – Durham, NH 03824

Cyanobacteria:
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Cyanobacteria are Earth’s oldest photosynthetic organisms
» Prokaryotic bacteria – not algae!
» Responsible for the oxygenation of our atmosphere
beginning 3.5 billion years ago

Research Questions:
1. To what degree does irrigation water with natural levels of
cyanobacteria and toxins contribute to the bioaccumulation of MC in food
plants? ~ Contamination via Direct Contact With Lake Water
2. To what degree do aerosols contribute to the bioaccumulation of MC
in food plants? ~ Contamination via Indirect Contact With Lake Water

Experimental Design:
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Use of hydroponics near a lake source to grow different
vegetable plants exposed to cyanobacteria directly at the
roots in the hydroponic system and/or indirectly via aerosols
Analysis of MC concentration via ELISA (Enzyme Linked
Immunosorbent Assay)

Previous Research: Aerosols
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Figure 1 – Microscopy photos of Oscillatoria limnetica and Microcystis aeruginosa - two
common toxin-producing cyanobacteria

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Well adapted to high-nutrient (eutrophic) systems but
common in low nutrient (oligotrophic) systems as well
» Often dominate in eutrophic systems
» Can form dense “blooms”, films, and scums

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MC found in aerosols from lakes with toxic cyanobacteria in
California5 & New Zealand6
Significant association found between non-alcoholic liver disease and
proximity to cyanobacterial blooms7
Methods for quantifying aerosolized cyanobacteria and toxins from
lakes developed8
Photo Credit:
Kate Langley

Figure 5 –
(C.L.A.M.) Field
instrument
designed to collect
aerosolized
particles emitted
from lake surfaces

Figure 9 – Lake Attitash, MA shoreline
with cyanobacteria bloom. Potential
study site for Summer 2016.

Implications:
Figure 6 – Epifluorescece Microscopy Image of an
aerosolized picocyanobacteria colony

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http://www.ibtimes.co.uk/dead-pigs-rivers-blood-shocking-photos-water-pollution-china-1459222

Figure 2 – Fishermen rowing through a
massive cyanobacteria bloom on Chaohu
Lake in Hefei, Anhui Province, in June, 2009

Figure 3 – Cyanobacteria float in Missisquoi
Bay of Lake Champlain in late August, 2015

Microcystin:
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Previous Research: Irrigation

http://www.burlingtonfreepress.com/story/life/2015/10/16/phosphorus-just-piece-blue-green-algae-puzzle/73394404

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Many cyanobacteria can produce an array toxins
Microcystin (MC) is the most common toxin produced
» Water soluble & highly stable
» Potent liver toxin and tumor promoter
» 76 dialysis patients died in 1996 after intravenous
exposure of MC contaminated water1
» Liver damage observed due to chronic exposure2
WHO Daily Intake Limits for Microcystin-LR only:
» 1 ug∙L-1 in drinking water & 0.04 ug∙kg-1 Body Mass3
MC has been found to accumulate in plant and animal tissues
including birds, fish, and invertebrates

Figure 4 – Healthy mouse liver (left) compared to mouse liver exposed to
Microcystin extract4

Pilot Study: MC accumulation seen in bush beans (Phaseolus
vulgaris) and cherry tomato plants using a known concentration of
Microcytis sp. for irrigation9
This chart right here
Figure 7 – MC levels found in
plants irrigated by contaminated
water sources extrapolated from
published MC concentrations.
Numbers reported are the total
MC that would be found in a
standard USDA ¼ cup serving of
vegetables compared to the
World Health Organization’s daily
intake limit for the weight of the
average US adult male (3.5µg d-1).

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Figure 8 – MC in the tissues of
wild blueberries collected July
2014 within 5 m of York Pond
and Christine Lake, NH.
Differences between berry
concentrations at the two
locations are significant
(p<0.05). One serving size of
blueberries of 200g (one cup)
would contain 0.89µg MC
(Christine Lake) and 0.76µg MC
(York Pond).12

Figure 10 – Example of potential hydroponic
system design. System would be placed near lake
shoreline for aerosol exposure, and irrigation
water would consist of contaminated lake water
for root exposure to MC.

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Add to the understanding of routes of exposure to
cyanotoxins, particularly aerosols
Help inform management decisions for irrigation sources and
overall water quality
» Agricultural irrigation ponds often provide ideal
habitat for cyanobacteria
» Due to warmer global temperatures and
anthropogenic eutrophication, large irrigation
sources such as the Great Lakes are also
experiencing increased cyanobacteria levels
Help inform lake management organizations and
communities on importance of reducing cyanobacteria levels
» Backyard vegetable gardens in lake communities
Provide information on sources of exposure to cyanotoxins
for humans and wildlife

Selected References:
1. Carmichael, WW, SMFO Azevedo, J Si An, RJR Molica, EM Jochimsen, S Lau, KL Rinehart, GR Shaw, and GK Eaglesham. 2001. Human fatalities from cyanobacteria: chemical and biological evidence
for cyanotoxins. Environmental Health Perspectives 109(7): 663-668.
2. Chen, J, P Xie, L Li, and J Xu. 2009. First identification of the hepatotoxic microcystins in the serum of a chronically exposed human population together with indication of hepatocellular damage.
Toxicological Sciences 108(1): 81-89.
3. Chorus, I. and J. Bartram [eds]. 1999. Toxic cyanobacteria in water: a guide to their public health consequences, monitoring, and management. World Health Organization and E & FN Spon,
London.
4. Briand, J.-F., S. Jacquet, C. Bernard, and J.-F. Humbert. 2003. Helth hazards for terrestrial vertebrates from toxic cyanobacteria in surface water ecosystems. Vet. Res. 34: 361-377
5. Backer, L., S. McNeel, T. Barber B. Kirkpatrick, C. Williams, M. Irvin, Y. Zhou, T. Johnson, K. Nierenberg, M. Aubel, R. LePrell, A. Chapman, A. Foss, S. Corum, V. Hill, S. Kieszak and Yung-Sung Cheng.
2010. Recreational exposure to microcystins during algal blooms in two California lakes. Toxicon 55: 909-921.
6. Wood, S. A. and D. R. Dietrich. 2011. Quantitative assessment of aerosolized cyanobacterial toxins at two New Zealand lakes. J. Environ. Monit. 13: 1617-1624.
7. Zhang, F., J. Lee, S. Liang, and C.K. Shum. 2015. Cyanobacterial blooms and non-alcoholic liver disease: evidence from a county level ecological study in the United States. Environmental Health.
14:41
8. Murby, A.L. and J.F. Haney. 2015. Field and laboratory methods to monitor lake aerosols for cyanobacteria and microcystins. Aerobiologia DOI 10.1007/s10453-015-9409-z
9. Lefebvre, B.R. 2013. The accumulations of the cyanobacterial toxin, microcystin, in cherry tomato (Solanum lycopersicum) and bush bean (Phaseolus vulgaris) plants. UNH Center for Freshwater
Biology Research 15(1): 1-11
10. Mohamed, ZA, and AM Al Shehri. 2009. Microcystins in groundwater wells and their accumulation in vegetable plants irrigated with contaminated waters in Saudi Arabia. Journal of Hazardous
Materials 172(2009): 310-315.
11. Codd, GA, JS Metcalf, and KA Beattie. 1999. Retention of Microcystis aeruginosa and microcystin by salad lettuce (Lactuca sativa) after spray irrigation with water containing cyanobacteria.
Toxicon 37(8): 1181-1185
12. Eisfeller, J. and A. Alexandrou. 2015. Microcystins in Vaccinium angustifolium from New Hampshire Lakes. University of New Hampshire URC. UNH, Durham, NH. Poster.

Acknowledgements:
A very special thanks to my advisor Dr. Jim Haney, my committee
members Jeff Schloss and Todd Guerdat, and to my lab-mate Kate Langley
for providing input and support in the continued development of my
research