Materials and Methods Sample Collection Process We collected surface water samples from the Raritan River (N = 5) between May 12 and August 6, 2016. Samples were collected under dry weather conditions. We waded into the middle of the river and held the manta trawl net (Fig. 2) perpendicular to the surface current flow for 15 minutes. Water column samples were collected using a manta trawl with a flow meter attached (Model 315, OceanTest, Inc.). The flow length multiplied by the width of the trawl provides the area sampled, allowing particle abundance per km2 to be calculated. Abstract Recent research indicates that plastic pollution is ex- tensive in freshwater systems. Wastewater treatment plants built decades ago were not designed to remove this newly emerging category of pollutant during wastewater treatment. Estabani & Fahrenfeld (2016) observed significant increases in microplastic densities in 125 – 250 and 250-500 mm size categories down- stream of four wastewater treatment plants located on the Raritan River. In an effort to determine the pres- ence and density of microplastics, and to assess poten- tial environmental effects associated with microplastic pollution in urban New Jersey drinking waters, we con- ducted research to collect and quantify surface water microplastics, identify organic compounds associated with these microplastic samples, and determine if living fish embryos were affected by exposure to the micro- plastics. We hypothesized that: 1) the density of micro- plastics would be highest in the heavily urbanized lower reaches of two New Jersey rivers; 2) persistent organic compounds would be associated with the recovered microplastic pollutants; and 3) exposure to these micro- plastics would result in observable physiological effects in juvenile zebrafish (Danio rerio). To test these hypoth- eses, we collected freshwater samples from five sites in the Raritan River watershed. Fig. 3 Filtering the collectec samples (left), preserving the collected sample (right). Microplastics in Urban Freshwaters: Conduit for Persistent Organic Pollution B. Ravit 1 *, C. Kooper 2 , G. Moreno 3 , B. Buckley 4 , H. Wang 4 , S. Meola 5 , D. Jones 6 , A. Hsieh 6 1 Department of Environmental Sciences, School of Environmental & Biological Sciences, Rutgers, The State University of New Jersey, New Brunswick, NJ, 2 Department of Biochemistry & Microbiology, School of Environmen- tal & Biological Sciences, Rutgers, The State University of New Jersey, New Brunswick, NJ, 3 Graduate Program in Microchemistry & Microbiology, Rutgers, The State University of New Jersey, New Brunswick, NJ, 4 Environ- mental & Occupational Health Sciences Institute, Rutgers, The State University of New Jersey, Piscataway, NJ, 5 NY/NJ Baykeeper, 52 W. Front St., Keyport, NJ, 6 Undergraduate Program in Environmental Sciences, School of Environmental & Biological Sciences, Rutgers, The State University of New Jersey, New Brunswick, NJ Fish Studies Pure plastic samples were purchased from Sigma Al- drich and were pulverized via dremel with a diamond tip followed by glass homogenization in 95% ethanol. The slurry was collected, centrifuged, and dried. The dried pulverized plastic was weighed and re- suspended in egg water to make microplastic concen- trations of 1 or 10μg/mL. The treatment schedule followed used AB strain Zebrafish embryos that were exposed to one type mi- croplastic per embryo. Twenty embryos were placed in glass vials and were treated with different concentrations of pure micro- plastic. Daily observations were made and lesions were rec- orded. Alcian Blue- Alizaran Red staining was done to help with photographs and measurements. Fig. 2 Passaic River sampling (2016) above the Dundee Dam. Fig. 4 (left) sample undergoing Fenton Reaction, (right) counting plastic density types and measuring sizes. Fig. 8 Zebrafish Exposure protocol. Density In the Rutgers University labs, large organic particles were rinsed with DI to collect any attached plastic par- ticles and the organic material was then discarded Following the protocols of Ericson et al. (2013) the re- maining sample was digested using the Fenton Reac- tion to remove remaining small organic particles Recovered microplastics were placed under a dissect- ing microscope and separated into one of three size categories and the type of plastic within each size cat- egory determined. Plastic density was calculated using the formula: Plastic density km-2 = # microplastics recovered di- vided by (net opening x flow distance) Microplastic size breakdowns were fairly evenly divid- ed between the 3 size factions (Table 1). The type of microplastic recovered varied by site (Table 2). The microplastic most frequently recovered was frag- ment, followed by foam > line > film > pellet (Table 2). References 1. Estahbanati, S., Fahrenfeld, N.L. 2016. Influence of wastewater treatment plant discharges on microplastic concentrations in surface water. Chemo- sphere 162:277-284. 2. Eriksen, M., Mason, S., Wilson, S., Box, C., Zellers, A., Edwards, W., Farley, H., Amato, S. 2013. Microplastic pollution in the surface waters of the Lauren- tian Great Lakes. Mar. Pollut. Bull. 77:177-182 Acknowledgements We thank NY/NJ Baykeeper for providing logistic assistance with sample collections. We thank the New Jersey Water Re- sources Research Institute (NJWRRI) for providing funding for this study. We gratefully thank Dr. S. Mason for her guidance and advice regarding sampling and analytic protocols. Fig. 7 Microplastic densities (plastic units km -2 ) observed in Raritan River surface waters under dry weather conditions in summer, 2016. 0.E+00 1.E+06 2.E+06 3.E+06 4.E+06 5.E+06 6.E+06 7.E+06 8.E+06 BRIDGEWATER RIVERSIDE BOYD PARK EDISON SAYREVILLE Raritan River Microplastic (Dry) Table 2. Recovered Microplastic Units by type Location Microplastic Type Total Fragment Pellet Line Film Foam Bridgewater 6 0 1 0 0 7 Piscataway 0 8 9 23 0 40 New Brunswick 7 1 6 0 0 14 Edison 13 1 39 11 5 69 Sayreville 8 0 2 5 34 49 Total Recovered 34 10 57 39 39 179 % Total Recovered 19% 6% 32% 22% 22% Table 1. Recovered Microplastic Units by size Location Microplastic Size Total 0.3-.99mm 1-4.75mm > 4.75mm Bridgewater 2 4 1 7 Piscataway 2 2 0 4 New Brunswick 5 9 0 14 Edison 19 22 30 71 Sayreville 7 30 12 49 Total Recovered 35 67 43 145 % Total Recovered 24.14% 46.21% 29.66% Next Steps Analyze microplastic samples using GC/MS to identify organic compounds adsorbed to the plastic Analyze plastic samples using pyrolysis to determine compounds contained in the plastic itself that may be released upon environmental exposure Collect 50 additional samples from 10 Raritan River locations during summer, 2017 Expose fish embryos to Raritan River samples. Fig. 1 Arial Map of a) Raritan and b) Passaic River Watershed. Sampling locations signified by blue dots. Location of facilities with NJPEDs discharge permits issued by the State of New Jersey identified by red dots. Results Microplastics were observed at all five sampled loca- tions in the Raritan (Figs 7; Table 1,2). Densities ranged from approximately 50,000 to over 1,000,000 microplastics km -2 (Fig. 7). Raritan microplastic density appeared to increase closer to Raritan Bay (Fig. 7). Fish embryos exposed to plastic standards exhibited abnormalities in total body length and pericardial sack size (Fig. 6) Fig. 6. Significant increase in total body length was seen in polyethylene HD 10 μg/mL treatment exposure. Significant increase in pericardial sack size was seen in polyethylene LD 10 μg/mL, pol- yethylene HD 1 μg/mL and 10 μg/mL, and polystyrene 1 μg/mL and 10 μg/mL treatment exposures. Fig 5. (left) fish pericardial sack, (right) total body length.
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Materials and Methods
Sample Collection Process
We collected surface water samples from the Raritan
River (N = 5) between May 12 and August 6, 2016.
Samples were collected under dry weather conditions.
We waded into the middle of the river and held the
manta trawl net (Fig. 2) perpendicular to the surface
current flow for 15 minutes.
Water column samples were collected using a manta
trawl with a flow meter attached (Model 315,
OceanTest, Inc.). The flow length multiplied by the
width of the trawl provides the area sampled, allowing
particle abundance per km2 to be calculated.
Abstract
Recent research indicates that plastic pollution is ex-
tensive in freshwater systems. Wastewater treatment
plants built decades ago were not designed to remove
this newly emerging category of pollutant during
wastewater treatment. Estabani & Fahrenfeld (2016)
observed significant increases in microplastic densities
in 125 – 250 and 250-500 mm size categories down-
stream of four wastewater treatment plants located on
the Raritan River. In an effort to determine the pres-
ence and density of microplastics, and to assess poten-
tial environmental effects associated with microplastic
pollution in urban New Jersey drinking waters, we con-
ducted research to collect and quantify surface water
microplastics, identify organic compounds associated
with these microplastic samples, and determine if living
fish embryos were affected by exposure to the micro-
plastics. We hypothesized that: 1) the density of micro-
plastics would be highest in the heavily urbanized lower
reaches of two New Jersey rivers; 2) persistent organic
compounds would be associated with the recovered
microplastic pollutants; and 3) exposure to these micro-
plastics would result in observable physiological effects
in juvenile zebrafish (Danio rerio). To test these hypoth-
eses, we collected freshwater samples from five sites
in the Raritan River watershed.
Fig. 3 Filtering the collectec samples (left), preserving the collected sample (right).
Microplastics in Urban Freshwaters: Conduit for
Persistent Organic Pollution B. Ravit1*, C. Kooper2, G. Moreno3, B. Buckley4, H. Wang4, S. Meola5, D. Jones6, A. Hsieh6
1Department of Environmental Sciences, School of Environmental & Biological Sciences, Rutgers, The State University of New Jersey, New Brunswick, NJ, 2Department of Biochemistry & Microbiology, School of Environmen-
tal & Biological Sciences, Rutgers, The State University of New Jersey, New Brunswick, NJ, 3Graduate Program in Microchemistry & Microbiology, Rutgers, The State University of New Jersey, New Brunswick, NJ, 4Environ-
mental & Occupational Health Sciences Institute, Rutgers, The State University of New Jersey, Piscataway, NJ, 5NY/NJ Baykeeper, 52 W. Front St., Keyport, NJ, 6Undergraduate Program in Environmental Sciences, School of Environmental & Biological Sciences, Rutgers, The State University of New Jersey, New Brunswick, NJ
Fish Studies
Pure plastic samples were purchased from Sigma Al-
drich and were pulverized via dremel with a diamond
tip followed by glass homogenization in 95% ethanol.
The slurry was collected, centrifuged, and dried. The
dried pulverized plastic was weighed and re-
suspended in egg water to make microplastic concen-
trations of 1 or 10μg/mL.
The treatment schedule followed used AB strain
Zebrafish embryos that were exposed to one type mi-
croplastic per embryo.
Twenty embryos were placed in glass vials and were
treated with different concentrations of pure micro-
plastic.
Daily observations were made and lesions were rec-
orded. Alcian Blue- Alizaran Red staining was done to
help with photographs and measurements.
Fig. 2 Passaic River sampling (2016) above the Dundee Dam.
Fig. 4 (left) sample undergoing Fenton Reaction, (right) counting plastic density types and measuring sizes.
Fig. 8 Zebrafish Exposure protocol.
Density
In the Rutgers University labs, large organic particles
were rinsed with DI to collect any attached plastic par-
ticles and the organic material was then discarded
Following the protocols of Ericson et al. (2013) the re-
maining sample was digested using the Fenton Reac-
tion to remove remaining small organic particles
Recovered microplastics were placed under a dissect-
ing microscope and separated into one of three size
categories and the type of plastic within each size cat-
egory determined.
Plastic density was calculated using the formula:
Plastic density km-2 = # microplastics recovered di-
vided by (net opening x flow distance)
Microplastic size breakdowns were fairly evenly divid-
ed between the 3 size factions (Table 1).
The type of microplastic recovered varied by site
(Table 2).
The microplastic most frequently recovered was frag-
ment, followed by foam > line > film > pellet (Table 2).
References 1. Estahbanati, S., Fahrenfeld, N.L. 2016. Influence of wastewater treatment
plant discharges on microplastic concentrations in surface water. Chemo-
sphere 162:277-284.
2. Eriksen, M., Mason, S., Wilson, S., Box, C., Zellers, A., Edwards, W., Farley,
H., Amato, S. 2013. Microplastic pollution in the surface waters of the Lauren-
tian Great Lakes. Mar. Pollut. Bull. 77:177-182
Acknowledgements We thank NY/NJ Baykeeper for providing logistic assistance
with sample collections. We thank the New Jersey Water Re-
sources Research Institute (NJWRRI) for providing funding for
this study. We gratefully thank Dr. S. Mason for her guidance
and advice regarding sampling and analytic protocols.
Fig. 7 Microplastic densities (plastic units km-2
) observed in Raritan River surface waters under dry weather
conditions in summer, 2016.
0.E+00
1.E+06
2.E+06
3.E+06
4.E+06
5.E+06
6.E+06
7.E+06
8.E+06
BRIDGEWATER RIVERSIDE BOYD PARK EDISON SAYREVILLE
Raritan River Microplastic (Dry)
Table 2. Recovered Microplastic Units by type
Location Microplastic Type Total
Fragment Pellet Line Film Foam
Bridgewater 6 0 1 0 0 7
Piscataway 0 8 9 23 0 40
New Brunswick
7 1 6 0 0 14
Edison 13 1 39 11 5 69
Sayreville 8 0 2 5 34 49
Total Recovered
34 10 57 39 39 179
% Total Recovered
19% 6% 32% 22% 22%
Table 1. Recovered Microplastic Units by size
Location Microplastic Size Total
0.3-.99mm 1-4.75mm > 4.75mm
Bridgewater 2 4 1 7
Piscataway 2 2 0 4
New Brunswick 5 9 0 14
Edison 19 22 30 71
Sayreville 7 30 12 49
Total Recovered 35 67 43 145
% Total Recovered
24.14% 46.21% 29.66%
Next Steps
Analyze microplastic samples using GC/MS to identify
organic compounds adsorbed to the plastic
Analyze plastic samples using pyrolysis to determine
compounds contained in the plastic itself that may be
released upon environmental exposure
Collect 50 additional samples from 10 Raritan River
locations during summer, 2017
Expose fish embryos to Raritan River samples.
Fig. 1 Arial Map of a) Raritan and b) Passaic River Watershed. Sampling locations signified by blue dots.
Location of facilities with NJPEDs discharge permits issued by the State of New Jersey identified by red dots.
Results
Microplastics were observed at all five sampled loca-
tions in the Raritan (Figs 7; Table 1,2). Densities
ranged from approximately 50,000 to over 1,000,000
microplastics km-2
(Fig. 7).
Raritan microplastic density appeared to increase
closer to Raritan Bay (Fig. 7).
Fish embryos exposed to plastic standards exhibited
abnormalities in total body length and pericardial sack
size (Fig. 6)
Fig. 6. Significant increase in total body length was seen in polyethylene HD 10 μg/mL treatment
exposure. Significant increase in pericardial sack size was seen in polyethylene LD 10 μg/mL, pol-
yethylene HD 1 μg/mL and 10 μg/mL, and polystyrene 1 μg/mL and 10 μg/mL treatment exposures.
Fig 5. (left) fish pericardial sack, (right) total body length.
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