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Mushrooms as Bioindicators of Heavy Metals in Sites Affected by Industrial Activity in the Macatawa Watershed

Hope College

Introductions

Amber BoschMajor: BiologyMinor: Environmental Science

Brooke MattsonMajor: GeologyMinor: Environmental Science

Kathleen FastMajor: GeologyMinor: Environmental Science

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MotivationTo investigate if the Lake Macatawa watershed contains regions of soils with elevated heavy metals using a cost and time-efficient methodology

Questions

Can mushrooms be used as bioindicators to assess heavy metal concentrations in the soil of the Lake Macatawa Watershed?

To what extent is biomonitoring able to accurately represent heavy metal concentrations from the environment?

https://www.visittheusa.com/destination/holland

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Biomonitoring

● Relatively cost effective method that employs natural organisms as a way of quantifying environmental conditions

● Some organisms and organic materials that have been used as biomonitors include oysters, bee honey, and various non-vascular plants

Why Mushrooms?

● Over 2,500 mushroom species in Michigan

● Found in a wide range of habitats● Underground structures allow for

bioaccumulation and depositing of metals into fruiting bodies

http://www.mushbox.co/The-Mushroom-Life-Cycle_b_2.html

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Industrial Activity & Metal Concentrations in Mushrooms

● Studies associated with industrial activity result in high metal concentrations within mushrooms

● Uptake amounts can vary in different species

● Overall trends make mushrooms adequate bioindicators

Sample Site Selection

2 Clean Sites:

● Upper Macatawa Natural Area● Hope College Nature Preserve

3 Contaminated Sites:

● Howard B. Dunton Park● Riley Trails● Dredge Material Placement Site

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Sample Sites

Sample Digestion

Clean & Dry Samples

8hrs 2X with HNO3 on hotplate

8hrs with HNO3 + H2O2 on hotplate

Heated into solution for 1hr

Dilute for analysis via Atomic Absorption

Homogenize samples and mass out ~0.5 grams

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Flame Atomic Absorption Spectroscopy

● Analysis of Pb, Cd, Ni, and Fe● Standards were tested between

0.05 - 5.0 ppm for Pb, Cd, and Ni 0.5 - 25 ppm for Fe ○ to account for the relatively high

environmental abundance in soil

Results: Iron RT3: 8 ppmRT6: 9 ppm

Du1: 87 ppm*Du2: 10 ppm

DSWT: 6 ppmDSY: 7 ppm**

UP5: 9 ppm

HCNP3: 9 ppmHCNP4: 9 ppm

R² = 0.99829*R² = 0.92536**R² = 0.99885

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Cadmium RT3: 2 ppmRT6: 2 ppm

Du1: 2 ppmDu2: 2 ppm

DSWT: 1 ppmDSY: 2 ppm

UP5: 5 ppm

HCNP3: 5 ppmHCNP4: 5 ppm

R² = 0.37682

Nickel RT3: 4 ppmRT6: 5 ppm

Du1: 4 ppmDu2: 5 ppm

DSWT: 3 ppmDSY: 6 ppm

UP5: 2 ppm

HCNP3: 2 ppmHCNP4: 2 ppm

R² = 0.94166

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Lead RT3: 1 ppmRT6: 1 ppm

Du1: 1 ppmDu2: 1 ppm

DSWT: 1 ppmDSY: 1 ppm

UP5: 1 ppm

HCNP3: 1 ppmHCNP4: 1 ppm

R² = 0.99744

Discussion

● No trend found between predicted contaminated and clean sites

● Considerations for high iron concentration of Du1● Of the four metals, iron was best suited for analysis with

instrumentation of this sensitivity● Variation in R2 values

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Discussion

Metal R2 ValueIron (0-50 ppm) 0.92536

Iron 2 (0-10 ppm) 0.99885

Iron (0-10 ppm) 0.99829Cadmium 0.37682Nickel 0.94166Lead 0.99744

Discussion

Experimental Concentration Average (ppm)

Literature Concentration Average (ppm)

Michigan Soils Concentration Average (ppm)

EPA Soil Contaminant Direct Contact Criteria (ppm)

Iron 17 70-4,000 10520 160,000

Cadmium 2 0.3-25 0.9 550

Nickel 5 0.6-18 12 40,000

Lead 1 1.0-8 9.2 400

EPA (2013), A.M et al. (2016), Chen et al. (2009), Zhu et al. (2011), Zhang et al. (2008), Siric et al. (2016)

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Sources of Error and Limitations

● Metal contamination due to lack of metal free zone in lab

● Varying and limited detection levels between metals for

Flame Atomic Absorption Spectrometry

● Results weakened by lack of consistent species and

small sample size

Future Research and Implications

● Within our study○ Use ICP-OES to analyze metal content

of Cd, Ni, and Pb○ Analysis of heavy metals in soil samples○ Sampling throughout the year

● Further experimentation of this biomonitoring methodology in the Macatawa Watershed and Midwest to determine viability

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Questions?

References

A, K., M, S., & K, S. (2016). Determination of heavy metals in edible mushrooms consumed in shahrekord. Journal of Shahrekord University of Medical Sciences, 18(1), 54-62.

Chen, X., Zhou, H., & Qiu, G. (2009). Analysis of several heavy metals in wild edible mushrooms from regions of china. Bulletin of Environmental Contamination and Toxicology, 83(2), 280-285.

EPA. (2013). Risk-Based Cleanup Criteria. Cleanup Criteria for Contaminated Soil and Groundwater, 11-20García, M., Alonso, J., Fernández, M. et al. Arch. Environ. Contam. Toxicol. (1998)Jarvie, M. (2017, April 13). Identifying wild Michigan mushrooms that are safe to eat. In Michigan State University Agriculture and Natural

Resources. Retrieved September 19, 2017, from http://msue.anr.msu.edu/news/identifying_wild_michigan_mushrooms_that_are_safe_to_eat

Radulescu, C., Stihi, C., Busuioc, G., Gheboianu, A. I., & Popescu, I. V. (2010). Studies concerning heavy metals bioaccumulation of wild edible mushrooms from industrial area by using spectrometric techniques. Bulletin of environmental contamination and toxicology, 84(5), 641-646.

Širić, I., Humar, M., Kasap, A., Kos, I., Mioč, B., & Pohleven, F. (2016). Heavy metal bioaccumulation by wild edible saprophytic and ectomycorrhizal mushrooms. Environmental Science and Pollution Research, 23(18), 18239-18252.

ZHANG Dan GAO Tingyan MA Pei LUO Ying SU Pengcheng. (2008). Bioaccumulation of heavy metal in wild growing mushrooms from liangshan yi nationality autonomous prefecture, china.

Zhu, F., Qu, L., Fan, W., Qiao, M., Hao, H., & Wang, X. (2011). Assessment of heavy metals in some wild edible mushrooms collected from yunnan province, china. Environmental Monitoring and Assessment, 179(1), 191-199. doi:10.1007/s10661-010-1728-5