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Effects of Oxybenzone Sunscreen on the Growth Of Lemna minor

Adrian Perono adrian.perono@ocvts.org
Marine Academy of Technology and Environmental Science

Introduction:
Oxybenzone, or benzophenone-3, is an active ingredient in many sunscreens. It is an
ultraviolet (UV) filter, meaning that it protects human skin against direct exposure to UV
radiation (Wang et al., 2016). It has proven to be an endocrine disruptor which is a chemical that
interferes with our hormones. Oxybenzone (OBZ) can enter rivers, lakes, groundwater, sewage,
and other areas through human activities and was identified as an “emerging environmental
pollutant” by the US Environmental Protection Agency. It contaminates kelp forests, seagrass
beds, and coral (Rodriguez, 2018). Recently, there have been many studies on the effects of OBZ
on coral. Whether in darkness or light, OBZ transformed the coral planulae from a motile state to
a deformed, sessile condition. Planulae exhibited an increasing rate of coral bleaching in
response to increasing concentrations of OBZ (Downs et al., 2015). Furthermore, OBZ has been
identified in various species of fish worldwide. After noticing this continuous pattern of toxicity
in animals and plants, the purpose of the project was to see how OBZ affects aquatic plants.
Lemna minor, or Duckweed, is a tiny floating plant that often forms a carpetlike cover on
the surface of the water and grows at a very fast rate. L. minor is important to the environment
because it can efficiently remove nitrogen, phosphorus, and heavy metals from water during
growth, and it has also been used in low-cost and low-energy wastewater treatment systems
(Ishizawa et al., 2017). Additionally, microbes that coexist with L. minor are believed to have
significant effects on growth in natural cultivation systems (Berg et al., 2014). Variable
fluorescence/maximum fluorescence (Fv/Fm) values measure chlorophyll fluorescence and
indicate plant health, with greater values reflecting better plant health and lower values reflecting
worse plant health. This study analyzed the effects that occurred when Lemna minor, or
duckweed, was exposed to different concentrations of OBZ-containing sunscreen.
Hypothesis:
By measuring the Fv/Fm levels, the plants exposed to OBZ will have lower Fv/Fm
values, as OBZ inhibits photosynthesis.
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Methods:
I. Plant Quantity and Usage
A. 150 plants of Lemna minor were planted
B. Live plants were bought from Carolina Biological Supply Company
C. All plants were kept at 20℃
D. They were grown using artificial sunlight
E. 10 plants were placed into each cup, for a total of 16 cups
II. OBZ Amounts
A. 16 cups were divided into 4 groups, each being exposed to a different amount of
sunscreen
a. Neutrogena Ultra Sheer® Dry-Touch Sunscreen Broad Spectrum SPF 100+
b. The plants were exposed to the sunscreen by a solution containing 1mL, 2
mL, or 4 mL of the sunscreen and 237 mL of water.
III. Testing
A. Leaf Chlorophyll levels were taken two or three times a week for eighteen days
using an AquaFluor™ Handheld Fluorometer in Fv/Fm.
IV. Site
A. Testing was done at Marine Academy of Technology and Environmental Science
in Stafford Township, New Jersey.
B. The growing of Lemna minor took place in the household.

Results:

The Fv/Fm values of all the plants in a concentration group were recorded and the daily
mean values were found and recorded in Table 1. Overall, the Fv/Fm values ranged from 0.5725
to 0.7233 during the eighteen day sampling period. This process was repeated for all four
concentrations of OBZ until all the mean values were found. None of the plants’ Fv/Fm values
varied greatly from the beginning to the end of the project (Figure 1). The data shows that the
plants exposed to 0 mL had higher Fv/Fm levels but only by a very small amount (Figure 2).
The data does not support the hypothesis that Lemna minor plants that are exposed to higher
concentrations of OBZ will have lowered Fv/Fm levels.
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Table 1: Daily mean Fv/Fm values for all concentration groups (0. 1. 2. 4 mL) across 18 days.
Values taken with AquaFluor™ Handheld Fluorometer. Calculated means are included (n=7)
Day 0 Day 4 Day 6 Day 8 Day 12 Day 14 Day 18 Mean
0 mL 0.645 0.6725 0.7175 0.71 0.69 0.7075 0.685 0.6896
1 mL 0.6075 0.6225 0.655 0.665 0.705 0.6975 0.65 0.6575
2 mL 0.5725 0.6825 0.665 0.68 0.705 0.6675 0.69 0.6662
4mL 0.6625 0.6575 0.7233 0.6767 0.6867 0.655 0.5575 0.6599

Figure 1: Daily mean Fv/Fm values for all concentration groups (0. 1. 2. 4 mL)
across 18 days. Values taken with AquaFluor™ Handheld Fluorometer.

Figure 2: Calculates Mean Fv/Fm Values [n=7 for each mean] for all
concentration groups (0. 1. 2. 4 mL) across 18 days with ± 5% Error Bars.
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Discussion:

These results suggest that Lemna minor has a strong ability to cope with higher
concentrations of OBZ in contaminated bodies of water since they were not greatly affected by
them. Another possible explanation for the outcomes of this project is that L. minor is a hardy
plant. For example, when the aquatic ecosystem dries out or declining temperatures occur, L.
minor has mechanisms to persist until conditions return that can support growth (DUCKWEED,
2019). Another example of this plant's hardiness is when it was exposed to pollutants like copper
and a chloroacetamide herbicide, it survived treatment very well, its growth was inhibited
scarcely, and no visible symptoms occurred (Obermeier et al., 2015). This would explain the lack
of change in terms of Fv/Fm values, because hardy plants tend to adapt better. One possible
explanation for the outcomes of this project is that L. minor has shown characteristics of an
accumulator species. An accumulator species is a species that gathers certain minerals or
nutrients from the soil and stores them in a more bioavailable form and in high concentration in
their tissues, then used as fertilizer or just to improve the mulch (Kourik & Kane, 2005). So, the
reason L. minor was able to cope with OBZ was because it stored the compound in a less-toxic
form, allowing the plant to survive. This brings up the possibility of the use of L. minor in
phytoremediation which refers to the use of plants rehabilitating water polluted by toxic metals
and organic compounds.
This project is important because it observes how L. minor would be affected in the wild
by the use of OBZ sunscreens. L. minor is an important source of food for fish and waterfowl
that provides a habitat for aquatic animals and yields enough shade to keep the growth of
oxygen-robbing algae down (Aquatic Biologists, Inc., 2020). Due to L. minor’s importance as a
species in aquatic food webs, it is of utmost importance to look out for changes in it’s health,
especially when caused by human activity.

Conclusion:

In conclusion, there is no trend between the concentration of OBZ that L. minor

plants are exposed to and lowered Fv/Fm levels. The hypothesis that the plants exposed
to OBZ will have lower Fv/Fm values was not supported, as none of the plants’ Fv/Fm
values varied greatly after being exposed to OBZ.

Acknowledgements:
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I would like to recognize my professors for providing the opportunity to conduct

independent research, as well as providing guidance throughout the process. I would
also like to thank Gabrielle Bonifacio for providing emotional support.
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References:
Berg, G., Grube, M., Schloter, M., & Smalla, K. (2014). Unraveling the plant microbiome:

looking back and future perspectives. Frontiers in Microbiology, 5.

https://doi.org/10.3389/fmicb.2014.00148

Downs, C. A., Kramarsky-Winter, E., Segal, R., Fauth, J., Knutson, S., Bronstein, O., Ciner, F.

R., Jeger, R., Lichtenfeld, Y., Woodley, C. M., Pennington, P., Cadenas, K., Kushmaro,

A., & Loya, Y. (2015). Toxicopathological Effects of the Sunscreen UV Filter,

Oxybenzone (Benzophenone-3), on Coral Planulae and Cultured Primary Cells and Its

Environmental Contamination in Hawaii and the U.S. Virgin Islands. Archives of

Environmental Contamination and Toxicology, 70(2), 265–288.

https://doi.org/10.1007/s00244-015-0227-7

DUCKWEED. (2019). Fao.org.

http://www.fao.org/ag/againfo/resources/documents/DW/Dw2.htm

Fern, K. (2021, April 22). Lemna minor - Useful Tropical Plants. Tropical.theferns.info.

http://tropical.theferns.info/viewtropical.php?id=Lemna+minor

Kourik, R., & Kane, M. (2005). Designing and maintaining your edible landscape naturally.

Chelsea Green Pub.

Lady Bird Johnson Wildflower Center - The University of Texas at Austin. (2016, September 27).

Www.wildflower.org. https://www.wildflower.org/plants/result.php?id_plant=lemi3

Obermeier, M., Schröder, C. A., Helmreich, B., & Schröder, P. (2015). The enzymatic and

antioxidative stress response of Lemna minor to copper and a chloroacetamide herbicide.

Environmental Science and Pollution Research, 22(23), 18495–18507.

https://doi.org/10.1007/s11356-015-5139-6
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Rodriguez, K. (2018). Sunscreen and Effects of Oxybenzone. Online Journal of Complementary

& Alternative Medicine, 1(1). https://doi.org/10.33552/ojcam.2018.01.000504

Wang, J., Pan, L., Wu, S., Lu, L., Xu, Y., Zhu, Y., Guo, M., & Zhuang, S. (2016). Recent

Advances on Endocrine Disrupting Effects of UV Filters. International Journal of

Environmental Research and Public Health, 13(8).

https://doi.org/10.3390/ijerph13080782