environments

Article
Quantifying Spatial and Temporal Trends of Microplastic
Pollution in Surface Water and in the Eastern Oyster
Crassostrea virginica for a Dynamic Florida Estuary
Linda J. Walters 1, * , Casey A. Craig 1,2 , Emily Dark 3,4 , Jessy Wayles 5 , Vincent Encomio 6,7 , Glenn Coldren 7,8 ,
Tess Sailor-Tynes 5 , David W. Fox 9 and Lei Zhai 9

1 Department of Biology, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816, USA
2 Florida Fish and Wildlife Conservation Commission, Fish and Wildlife Research Institute, 100 Eighth Ave SE,
St Peterburg, FL 33701, USA
3 Florida Department of Environmental Protection-Indian River Lagoon Aquatic Preserves, 3300 Lewis St.,
Fort Pierce, FL 34981, USA
4 Martin County Board of County Commissioners, 2401 SE Monterey Rd., Stuart, FL 34996, USA
5 Marine Discovery Center, 520 Barracuda Blvd, New Smyrna Beach, FL 32169, USA
6 UF IFAS, Florida Sea Grant Martin County Extension, 2014 SE Dixie Highway, Stuart, FL 34996, USA
7 Florida Oceanographic Society, 890 NE Ocean Blvd., Stuart, FL 34996, USA
8 Department of Biology, Indian River State College, 3209 Virginia Ave., Fort Pierce, FL 34981, USA
9 Department of Chemistry and Nanoscience Technology Center, University of Central Florida, 4000 Central
Florida Blvd., Orlando, FL 32816, USA
* Correspondence: [email protected]; Tel.: +1-407-823-2148

Citation: Walters, L.J.; Craig, C.A.; Abstract: Microplastics (MPs) are a ubiquitous pollutant, emphasizing the need to understand
Dark, E.; Wayles, J.; Encomio, V.; their abundance and the factors that influence these patterns around the globe. In a prior study,
Coldren, G.; Sailor-Tynes, T.; Fox, high numbers of MPs were found in surface waters and tissues of the oyster Crassostrea virginica
D.W.; Zhai, L. Quantifying Spatial
collected from one location in the Indian River Lagoon (IRL, FL, USA). To better understand spatial
and Temporal Trends of Microplastic
and temporal variability of MPs throughout the IRL, for one year, monthly surface water samples
Pollution in Surface Water and in the
were collected from 35 sites, while oysters were collected quarterly from 12 sites. Microscopy
Eastern Oyster Crassostrea virginica
and ATR-FTIR were used to quantify MP. In total, 3755 MPs were found in 44% of water samples
for a Dynamic Florida Estuary.
Environments 2022, 9, 131.
(mean density ± CI: 1.47 ± 0.09 MP/L). South IRL water had the most MPs, likely associated with
https://doi.org/10.3390/ proximity to urbanization, inlets (MP sinks) and tributaries (MP sources). MP (n = 3181) were found
environments9100131 in 70% of examined C. virginica (n = 1402). Abundances of MP in oysters were lower in the spring
and in north IRL. The overall mean abundance was 2.26 ± 0.16 MP/oyster, and the density was
Academic Editor: Ioannis K.
2.43 ± 0.52 MP/g wet tissue weight. Our results provide a more complete picture of MPs in the IRL,
Kalavrouziotis
a subtropical, shallow-water estuarine system.
Received: 25 August 2022
Accepted: 10 October 2022 Keywords: citizen-science; Indian River Lagoon; microfiber; Mosquito Lagoon; shellfish restoration
Published: 13 October 2022

Publisher’s Note: MDPI stays neutral

with regard to jurisdictional claims in
published maps and institutional affil- 1. Introduction
iations. Plastic is both a common household material and pervasive pollutant despite its rela-
tively short history (e.g., [1–3]). Synthetic plastic was first created by Leo Baekeland in 1907;
mass production of plastic, however, did not begin until the 1950s when a new generation
of plastics (e.g., PVC, polyvinyl chloride; PS, polystyrene; Nylon; PE, polyethylene; PP,
Copyright: © 2022 by the authors.
polypropylene; and PET, polyethylene terephthalate) made this feasible [4,5]. Global plastic
Licensee MDPI, Basel, Switzerland.
production continued to increase, with an estimated 8300 million metric tons (Mt) produced
This article is an open access article
up to 2015; 79% is now in landfills or the environment, 9% has been recycled, and 12% was
distributed under the terms and
incinerated [6]. In 2015, an estimated 4.8 to 12.7 Mt of plastic debris entered Earth’s oceans
conditions of the Creative Commons
Attribution (CC BY) license (https://
from a myriad of sources, including ship overspill, container wash-off, coastal development
creativecommons.org/licenses/by/
and litter, with a tenfold increase predicted by 2025 [7,8].
4.0/).

Environments 2022, 9, 131. https://doi.org/10.3390/environments9100131 https://www.mdpi.com/journal/environments

Environments 2022, 9, 131 2 of 18

Once in the marine environment, plastics are subject to solar, thermal, mechanical
and biodegradation, which can weaken or fragment the plastics into smaller dimensions
called microplastics (MPs) [9,10]. The National Oceanic and Atmospheric Administration
defines an MP as any plastic object less than or equal to 5 mm in size, but can be further
categorized into two subclasses, primary and secondary MPs [9]. Primary MPs are those
that are manufactured at a small size, whereas secondary MPs form via fragmenting from
a larger plastic object [11]. Primary MPs include microbeads in personal care products
and ‘nurdles’, raw material formed into small pellets for easy transport that are used to
make larger plastic items [12,13]. Secondary MPs include fibers, fragments, foams, and
films which vary in shape, depending on how they are formed [11]. Oceanic MPs are
predominantly textile fibers (35%), fragments associated with city dust (24%), pieces of tires
(28%), or nurdles/beads (3% [7]). Fibers are especially common in estuaries and coastal
waters [14]. For example, Simon-Sanchez et al. [15] found fibers were the most abundant
type (70%) of MP in the Ebro Delta estuary in Spain. Luo et al. [16] documented similar
dominance of microfibers (80%) in coastal waters in the Shanghai area and found that MP
abundance increased closer to the city.
Plastic ingestion in marine biota has been documented in hundreds of species at
varying trophic levels (e.g., [17,18]). Species of particular interest are filter-feeders such
as oysters, clams, and mussels [14,19–21]. The eastern oyster, Crassostrea virginica, is an
estuarine species along the western Atlantic seaboard known to ingest MPs and negative
impacts on its physiology have been reported [22]. To feed, many shellfish species extract
particulate matter from the water, encase it in mucous, and then either reject or digest
the particles. If rejected, the material is excreted as pseudofeces. If accepted, the material
is brought to the mouth opening, passed through the digestive tract, and any remainder
is excreted as feces [23]. MPs have been found in both the feces and pseudofeces of
C. virginica [21]. Additionally, MP ingestion in Crassostrea has been documented to reduce
reproductive success (e.g., [24]) as well as reduce growth and survival [22].
Microplastics are a ubiquitous pollutant in the marine environment and a potential
risk to marine biota, emphasizing the need to understand MP abundance and the factors
that influence these patterns in marine systems. Microplastics are transported through
coastal systems by a dynamic series of forces such as rain, wind, freshwater discharge,
waves, tides, salinity gradients, surface drift, biofouling, and storm events [25,26]. All
of these variables can create seasonal trends in MP abundances (e.g., [27]). Identifying
what factors influence MP abundances in hydrologically complex coastal landscapes is a
defined research gap in the MPs field [28]. Our research goal is to understand spatial and
temporal patterns of MPs in surface waters of the Indian River Lagoon system that spans
251 km along the east coast of central Florida, USA. Additionally, we examine spatial and
temporal patterns of MPs in tissues of C. virginica to better understand how this keystone,
filter-feeding species has been impacted by anthropogenic MP pollution. To broaden our
impact on the IRL community, we worked with citizen-scientists along the length of the
IRL to collect and process samples after extensive training.

2. Methodology
2.1. Study Location
Our focal area is Florida’s largest lagoon, the Indian River Lagoon (IRL), which spans
40% of Florida’s east coast from Ponce de Leon Inlet in the north (29.075898◦ N, 80.917571◦ W)
to Jupiter Inlet in the south (26.944768◦ N, 80.073952◦ W). The IRL is a bar-built lagoon that
has limited water exchange through five inlets and is hydrologically complex [29–31]. Four
of the five inlets within the lagoon, Ponce de Leon, Sebastian, Ft Pierce, and St Lucie, were
included within the defined boundaries of this study. The IRL falls within the boundaries of
Volusia, Brevard, Indian River, St. Lucie, Martin, and Palm Beach counties. IRL water (hereafter
lagoon water) flows through three interconnected water bodies: Mosquito Lagoon, Indian
River, and Banana River, has an average depth of 1.2 m [29]. Saltwater influx comes from the
Atlantic Ocean through inlets, while freshwater input is predominantly from rainfall, discharge,
 Environments 2022, 9, x FOR PEER REVIEW 3 of 19

IRL water (hereafter lagoon water) flows through three interconnected water bodies: Mos-
Environments 2022, 9, 131
quito Lagoon, Indian River, and Banana River, has an average depth of 1.2 m [29]. Saltwa- 3 of 18
ter influx comes from the Atlantic Ocean through inlets, while freshwater input is pre-
dominantly from rainfall, discharge, and runoff from nearby land [29]. Currents, tides,
and circulation patterns are influenced by factors dependent on location from the nearest
and runoff from nearby land [29]. Currents, tides, and circulation patterns are influenced by
inlet; areas closer to inlets have a larger tidal influence while stretches between inlets are
factors dependent on location from the nearest inlet; areas closer to inlets have a larger tidal
primarily driven by wind and freshwater input [29]. This research is focused on IRL oys-
influence while stretches between inlets are primarily driven by wind and freshwater input [29].
ters and their surrounding waters because C. virginica has long been harvested for human
This research is focused on IRL oysters and their surrounding waters because C. virginica has
consumption and there are long-term efforts to restore these oyster populations (e.g., [32])
long been harvested for human consumption and there are long-term efforts to restore these
for the ecological benefits they provide for fishes [33,34], wading birds [35], and aquatic
oyster populations (e.g., [32]) for the ecological benefits they provide for fishes [33,34], wading
invertebrates [36,37].
birds [35], and aquatic invertebrates [36,37].
2.2.Sample
2.2. SampleCollection
CollectionandandCitizen
CitizenScience
Science
Lagoonwater
Lagoon waterwaswascollected
collectedfrom
fromthetheIRL
IRLover
overaa12-month
12-monthsampling
samplingperiod,
period,between
between
March 2019 and February 2020. Water was collected once per month
March 2019 and February 2020. Water was collected once per month from 35 sites that from 35 sites that
extendedthe
extended thelength
lengthof ofthe
theIRL.
IRL.All
Allsites
siteswere
wereaccessible
accessiblefrom
fromshore
shoreand andononpublic
publiclands
lands
(Figure1,1,Table
(Figure Table1).1).Each
Eachmonth,
month,lagoon
lagoonwater
watersamples
samplesfromfromall
allsites
siteswere
werecollected
collectedwithin
within
aa4-day
4-daytime
timeperiod
periodtotolimit
limittemporal
temporalvariation.
variation. At
At each
each site,
site, five
five replicate
replicate 11LLsurface
surface
lagoon water samples were collected using a discrete sampling protocol
lagoon water samples were collected using a discrete sampling protocol [38,39]. Sample [38,39]. Sample
bottleswere
bottles weretriple-rinsed
triple-rinsedin in0.45
0.45µmµmfiltered
filtereddeionized
deionizedwater
waterininthethelaboratory,
laboratory,andandthen
then
againwith
again withlagoon
lagoonwater
waterupon
uponsitesitearrival
arrivaltotoremove
removeany anyexisting
existingcontamination.
contamination.Bottle
Bottle
rinsingoccurred
rinsing occurredatat least
least 10 10 m away
m away fromfrom the sample
the sample collection
collection location.
location. Rinsed Rinsed
bottlesbottles
were
were partially
partially submergedsubmerged to collect
to collect the topthe
5 cm topof5lagoon
cm of lagoon
water andwater
cappedand capped while sub-
while submerged.
merged.
At At each
each site, site, parameters
abiotic abiotic parameters of airwater
of air and and water temperature
temperature werewere recorded
recorded using
using a
a thermometer ◦ (°C), salinity using a refractometer (ppt), and mean
thermometer ( C), salinity using a refractometer (ppt), and mean wind speed using an wind speed using an
anemometer (km/h).
anemometer (km/h). Samples
Sampleswere weretransported
transported back to laboratories
back to laboratories andand
keptkept
at room tem-
at room
perature (20(20
temperature °C)◦through
C) through completion
completionof processing.
of processing.

Figure1.1.Indian
Figure Indian River
River Lagoon
Lagoon microplastic
microplastic water
water sampling
sampling sites (points)
sites (points) andreef
and oyster oyster
areasreef areas
(boxes).
(boxes).
Environments 2022, 9, 131 4 of 18

Table 1. Indian River Lagoon water sampling sites from north to south.

Site No. Site Name Abbreviation Region Latitude Longitude

1 Smyrna Dunes Park SDP N 29.063822 −80.915744
2 Marine Discovery Center MDC N 29.030158 −80.917641
3 River Breeze Park RBP N 28.898601 −80.85174
4 CANA Boat Ramp CANAB N 28.934251 −80.829475
5 CANA Parking Lot #5 CANA5 N 28.857672 −80.777248
6 Haulover Canal HOC N 28.706285 −80.720657
7 Parrish Park PPK N 28.623625 −80.794767
8 Campground CAMP N 28.504 −80.7801
9 Briarwood BW N 28.42123 −80.75245
10 Lee Wenner Boat Ramp LWBR N 28.355086 −80.722994
11 Rockledge ROCK N 28.3014 −80.7005
12 Rotary Park RPK N 28.2295 −80.6714
13 Pineapple PINE C 28.154 −80.6382
14 Front Street FS C 28.079558 −80.599847
15 Malabar MAL C 27.9862 −80.5532
16 Christensen CHR C 27.93112 −80.526022
17 Outriggers OUT C 27.855367 −80.492992
18 Sebastian SEB C 27.80892 −80.466215
19 Environmental Learning Center ELC C 27.758069 −80.415706
20 Vero VERO C 27.654303 −80.368983
21 Round Island RI C 27.561131 −80.328635
22 Wildcat WC C 27.495292 −80.303114
23 Bear Point BP S 27.429391 −80.281382
24 Midway MID S 27.38723 −80.297868
25 Jensen Beach JEN S 27.308302 −80.22226
26 Palm City Bridge PCB S 27.155333 −80.261
27 Riverwalk RW S 27.20225 −80.253883
28 Fish House FH S 27.151083 −80.199867
29 Twin Rivers TR S 27.164933 −80.18215
30 Driftwood DW S 27.255533 −80.23085
31 Jensen Beach Impound JBI S 27.260117 −80.209233
32 River Cove RC S 27.21435 −80.183983
33 House of Refuge HOR S 27.199617 −80.166283
34 Indian Riverside Park IRP S 27.228535 −80.212716
35 Jimmy Graham Boat Ramp JGBR S 27.09958 −80.145616

Citizen scientists associated with the University of Central Florida (UCF; current
students and alumni) and partnering non-profit conservation agencies situated along
the IRL assisted with water sample collection and microscopic identifications. Citizens
who ranged in age from 16 to 80 were recruited through existing agencies’ volunteer
networks, social media postings, and by word of mouth. To be included as a citizen
scientist, individuals had to be available for the 12 months of the project and have their
own transportation. Citizen recruits underwent extensive MP training where they were
educated about MP generation and pollution, scientific procedures used for field sampling
of MP, and sample inspection protocols in the laboratory. Once successfully trained, citizen
scientists were independently deployed to collect water samples each month at pre-selected
sites that were near their homes. Lagoon water samples were processed and inspected for
MPs in laboratories at UCF or the associated conservation agencies under direct supervision
of university/agency investigators. A minimum of 10% of all samples were then re-checked
by university personnel as required by our EPA QAPP (Quality Assurance Project Plan).
On all sample collection and processing dates, all participants were requested to wear only
natural fiber clothing.
To compare to the collected water samples, the oyster C. virginica was collected quar-
terly for one year from 12 intertidal reefs in the IRL. This included 6 reefs from the north,
3 from the central, and 3 from the south IRL (Figure 1). Sampling reefs in the north region
were randomly selected using a random number generator (www.random.org) (accessed
on 1 December 2018), while central and southern reefs were the only sustainable, inter-
tidal reefs accessible in each respective region (E. Dark, pers. comm.; [40]). Additionally,
the 3 southern reefs were part of oyster restoration efforts. Sampling distribution was
skewed to the north to be representative of C. virginica abundance in the IRL, as there is
Environments 2022, 9, 131 5 of 18

a historical downward trend in oyster abundance as latitude decreases [40,41]). At each

reef, 30 individual C. virginica were collected. Fifteen large (shell length ≥ 36 mm) and
fifteen small (shell length < 36 mm) C. virginica were haphazardly collected from each
reef, wrapped in aluminum foil, bagged, and placed on ice. Oysters from all 12 reefs were
collected within a 7-day window to limit temporal variation. Samples were brought to the
University of Central Florida Department of Biology laboratory for storage in a −20 ◦ C
freezer until processing.

2.3. Sample Processing

IRL water samples were vacuum-filtered at room temperature using Whatman ni-
trocellulose membrane filter paper (47 mm, 0.45 µm pore size) to extract MP, and placed
in triple-rinsed, sealed, 60 × 15 mm Petri dishes. Filters were inspected once dry using
a dissecting microscope (Leica EZ4, Morrisville, NC, USA) at 20×–40× magnification.
MP type, color, and size (mm) were recorded following protocol established by the Shaw
Institute [42]. To distinguish between natural and synthetic items, potential MPs were
prodded using forceps to test breakage, and examined for discrete variation in color, shape,
and margins (smooth, jagged, frayed) along their lengths.
Individual C. virginica were thawed and shell heights (mm) were recorded using
calipers. After each oyster was shucked, the blotted-dry, wet-weight of soft tissue weighed
(g) was determined using a top-loading balance (Ohaus Scout Pro, Parsippany, NJ, USA),
and then placed in individual glass Erlenmeyer flasks (125 mL for small, 250 mL for
large oysters). Digestion protocol followed procedures established by [43] for the optimal
extraction of MPs from bivalve tissues. A 10% potassium hydroxide (KOH) solution was
added to each flask at a ratio of 3:1 volume (mL) to wet tissue weight. Flasks were covered
and placed in a shaking incubator at 40 ◦ C at 60 rpm for 24 h, and then removed and
left at room temperature for an additional 24 h where tissue digestion was completed.
A 1.0 M citric acid solution was added to the digested tissue solution until a neutral pH
(7.0) was reached to prevent an interaction with filters [20]. The neutralized solution was
vacuum-filtered under a fume hood using Whatman glass microfiber filters (90 mm, 1.2 µm
pore size) and placed in triple-rinsed Petri dishes for later quantification.

2.4. Limiting Polymer Contamination and Degradation

Procedural MP contamination was controlled for by triple-rinsing all equipment used
during digestion and filtration with 0.45 µm filtered deionized (DI) water prior to each
use (M.M. Patterson, pers. comm.). Solutions used during digestions were also made with
0.45 µm filtered DI water. Chemical digestion of oysters was conducted in a fume hood to
prevent polymer contamination during the filtration process [44]. KOH was preferrable
to digest bivalve soft tissue and extract MP particles as small as 1 µm in size because it
preserves major polymers, including rayon [43].
Aerial contamination was quantified during microscopy by using five filter-control
blanks (filters dampened with 0.45 µm filtered, deionized water placed in triple-rinsed Petri
dishes) [44,45]. Blanks (exposed filters) were haphazardly placed on the table immediately
around the microscopy station at all times during inspection to quantify potential air
contamination while samples were exposed [45]. Blanks were inspected for MP, then
normalized (AbundanceN ) to a mean contamination rate per minute.

2.5. Fourier-Transform Infrared Spectroscopy

To supplement MP identification, polymer composition was determined using attenu-
ated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR) at the University
of Central Florida Nanoscience Technology Center using a Shimadzu IRSpirit-T instrument.
A subset of samples containing MPs (10% of water and 10% of C. virginica samples with
MPs) were randomly selected (www.random.org) (accessed on 1 March 2020). And all
potential MPs larger than 0.5 mm in size were scanned [46]. MPs were scanned in the
600 to 4000 cm−1 range and spectra were matched to the reference library from Shimadzu
Environments 2022, 9, 131 6 of 18

using differential derivative point matching (ATR-FTIR Polymer and Polymer Additives
Database #220-93143-07, 2020). A score, also known as hit quality index (HQI), for each
spectrum was calculated to measure percent match using the equation:
 1 
D 3
1− S +1
HQI = (1)
2
where D is the summation of the primary and reference spectra by calculation of a fitting
method, and S is the area of the primary derivative curve of the sample spectrum. Spectra
were automatically included as a polymer if score match was 700 or higher, ambiguous
scores of 600–700 were manually sorted for inclusion in analysis, and scores of 600 or below
were excluded [2,47]. Ambiguously scored spectra were inspected and differentiated by
visual peak matching. Polyester is predominantly PET and could not be elucidated as a
distinctly different polymer, so polyester signals were classified as PET [6]. A subset of
signals (10%) on MP identified in control blanks were scanned using ATR-FTIR to determine
any overlap between polymers found in the IRL and aerial contamination.

2.6. Statistical Methods

Abundance data for MP in lagoon water and C. virginica MPs were broadly dispersed
and had a high presence of zeros, so each were analyzed using negative binomial gen-
eralized linear modelling (GLM) for zero-inflated data (R package “pscl”). To quantify
spatial and temporal variations in MP abundance in lagoon water, predictor variables tested
included region, site, and season. Seasons were defined by standard meteorological seasons
in Florida (Spring = March–May, Summer = June–August, Fall = September–November,
Winter = December–February). To determine what factors may influence MP abundance
in water, distance to the nearest tributary (km) and distance to the nearest inlet (km) were
tested as predictor variables in models. Distinct regional differences in MP abundance were
apparent, so both whole-IRL and independent regional model analyses were incorporated
to distinguish trends more precisely. The IRL is very expansive; therefore, models with
predictors of distance to an inlet and tributary were only included within the individual
regional analyses.
Control blanks were normalized to a contamination rate per minute (CM) using the formula:

MPB
CM = (2)
TB

where MPB is the mean number of MP per blank and TB is the time that blanks were
exposed in minutes [21]. Contamination per minute values then were used to calculate the
contamination per sample (CS ) using the formula:

CS = C M × TE (3)

where TE is the length of time each filter was exposed during inspection [21]. Contamination
per sample values were incorporated as a covariate in water models but only included if
significant in the model.
To quantify MP abundance and fluctuations in IRL oysters, predictor variables tested
in models included region, site, season, and shell height. To determine what factors may
influence MP abundance in oysters, distance to the nearest tributary, and distance to the
nearest inlet were also tested as predictor variables. Contamination per sample and tissue
weight were incorporated in all oyster models but only included in analyses if significant.
Post hoc model selection using all possible predictor variable combination for Akaike
information criterion (AIC) was used to determine which variables best predicted MP
abundance in both lagoon water and oyster samples from the IRL. Regional differences in
MP abundance were apparent so both whole-IRL and independent regional model analyses
 influence MP abundance in oysters, distance to the nearest tributary, and distance to the
nearest inlet were also tested as predictor variables. Contamination per sample and tissue
weight were incorporated in all oyster models but only included in analyses if significant.
Post hoc model selection using all possible predictor variable combination for Akaike infor-
mation criterion (AIC) was used to determine which variables best predicted MP abun-
Environments 2022, 9, 131 7 of 18
dance in both lagoon water and oyster samples from the IRL. Regional differences in MP
abundance were apparent so both whole-IRL and independent regional model analyses
of C. virginica were incorporated. Regional models used the same predictor variables as
of C.whole-IRL
the virginica were incorporated.
models Regional
to distinguish models used the same predictor variables as the
trends.
whole-IRL
Linearmodels to distinguish
regressions were usedtrends.
to determine differences in air and water temperature
Linear
between IRLregressions were
regions, and used to
seasons. determine were
Regressions differences in air
also used to and water temperature
determine wind speed
between IRL regions, and seasons. Regressions were also used to determine
and salinity differences between IRL regions, sites, and months. All statistical wind speed
analyses
and salinity differences between IRL
were performed using R, version 4.0.3 [48]. regions, sites, and months. All statistical analyses
were performed using R, version 4.0.3 [48].
3. Results
3. Results
3.1.
3.1. Microplastics
Microplastics inin Lagoon
Lagoon Water
Water
Overall,
Overall, a total of 3755 MPs
a total of 3755 MPs were
were observed
observed in in 44%
44% ofof all
all lagoon
lagoon water
water samples.
samples. WhenWhen
separated
separated by regions, 70.7% of north IRL samples contained MP, while 29.6% and
by regions, 70.7% of north IRL samples contained MP, while 29.6% and 66.0%
66.0%
of
of samples contained MP for the central and south regions, respectively. These were
samples contained MP for the central and south regions, respectively. These data data
collected by 84 by
were collected citizen-scientists who collectively
84 citizen-scientists who collectivelydonated 1600 h1600
donated to this MP
h to water
this MP effort.
water
Lagoon water MP
effort. Lagoon waterdensity ranged from
MP density ranged 0 to 25.00MP/L
from and
to 25.0 had aand
MP/L meanhad(±CI)
a mean of 1.47 ± 0.09
(±CI) of
1.47 ± 0.09 MP/L (Figure 2). Mean normalized abundance reduced the range slightlyMP
MP/L (Figure 2). Mean normalized abundance reduced the range slightly to 0–24.6 to
and theMP
0–24.6 meanandtothe
1.42 ± 0.10.
mean to MP
1.42abundance
± 0.10. MPdiffered
abundance between the IRL
differed regions.
between the Central sites
IRL regions.
had the lowest
Central sites hadMPthe abundance,
lowest MP followed by thefollowed
abundance, north, thenby south regions
the north, then(p south
< 0.001regions
for all,
Figure
(p 2). Fibers,
< 0.001 fragments,
for all, Figure films, and
2). Fibers, foams were
fragments, films,found in water,
and foams werewith fibers
found in comprising
water, with
95.6% MP. Fragments,
fibers comprising 95.6%foams, and films foams,
MP. Fragments, comprised the remaining
and films comprised 3.9%, 0.3%, and 3.9%,
the remaining 0.2%
MPs, respectively. Plastics ranged in size from 0.1 mm to 30.0 mm and
0.3%, and 0.2% MPs, respectively. Plastics ranged in size from 0.1 mm to 30.0 mm and hadhad a mean longest
dimension
a mean longest(±CI)dimension
of 1.9 ± 0.1(±
mm.CI) of 1.9 ± 0.1 mm.

Figure 2. Microplastic abundance per liter of water from the north, central, and south Indian River
Lagoon. Valuesare
Lagoon. Values aremean
meanabundance
abundance (point)
(point) andand 95%
95% confidence
confidence interval
interval of mean.
of the the mean. (GLM,
(GLM, p<
p < 0.05,
0.05, North = 716, Central = 598, South
North = 716, Central = 598, South = 776).= 776).

The most plausible model to predict lagoon-wide MP abundance in IRL water included
site and season as predictor variables (AIC = 6283.6; McFadden pseudoR2 = 0.12, Table 2).
For the entire IRL and the north IRL, MP abundance was higher in lagoon water in fall
and winter (p ≤ 0.02). In central and south regions, the most plausible predictors of MP
abundance were site and season (AIC = 1131.2, McFadden pseudoR2 = 0.10, Table 2) and
(AIC = 2710.5; McFadden pseudoR2 = 0.12), respectively (Table 2). Abundance was also
higher in fall and winter (p < 0.001) for both regions. When the presence/absence of MPs
was considered for IRL water overall, winter was a significant predictor of MP presence
(p = 0.005). Despite this lagoon-wide pattern, MP presence was not predicted by any season
when regions were considered separately.
Environments 2022, 9, 131 8 of 18

Table 2. Zero-inflated negative binomial GLM models of MP abundance in lagoon water overall
and by region (north, central, south). Values reported are AIC, delta AIC, degrees of freedom, and
AIC weight.

Indian River Lagoon Water AIC ∆AIC df AIC Weight

Site + season 6283.6 0 77 1
Site 6306.6 23 71 <0.001
Tributary + region + season 6569.9 286.3 15 <0.001
Tributary + region 6584.1 300.5 9 <0.001
Region + season 6644.6 361 13 <0.001
Region 6660.2 376.6 7 <0.001
Season 6938.3 654.8 9 <0.001
Inlet 6946.8 663.2 7 <0.001
Tributary 6951.4 667.8 5 <0.001
North Lagoon AIC ∆AIC df AIC Weight
Season 2422.9 0 9 0.8221
Site + season 2426.0 3.1 31 0.1748
Site 2434.1 11.2 25 0.0031
Central Lagoon AIC ∆AIC df AIC Weight
Site + season 1131.2 0 27 0.7207
Site 1133.1 1.9 21 0.2771
Inlet + season 1143.6 12.4 11 0.0015
Inlet 1145.0 13.8 5 <0.001
Tributary 1179.1 47,9 5 <0.001
Season 1209.9 78,7 9 <0.001
South Lagoon AIC ∆AIC df AIC Weight
Site + season 2710.5 0 33 1
Site 2742.7 32.2 27 <0.001
Tributary + season 2912.9 202.5 11 <0.001
Tributary 2920.5 210 5 <0.001
Season 3003.5 293 9 <0.001

MP abundances differed between IRL regions with regard to oceanic and freshwater
influences. In the north IRL, MP abundance did not vary with distance to a tributary or
inlet. There are no tributaries in the north and one oceanic inlet (Ponce de Leon Inlet).
A different trend was apparent in the central IRL, where MP abundance decreased with
increasing distance from a tributary (p < 0.001) and increased with increasing distance from
an inlet (p = 0.001). Abundance decreased by 0.99 MP/L for every 1 km increase in distance
from a tributary and increased by 0.82 MP/L for every 1 km increase in distance from an
inlet in the central IRL. In the southern region, no oceanic influence was apparent; however,
there was tributary influence and MP abundance decreased by 0.88 MP/L for every 1 km
increase in distance from a tributary (p < 0.001).

3.2. Microplastics in Oysters

Crassostrea virginica (n = 1402) from the IRL contained a total of 3181 MPs. The
composition of MP was dominantly fibers (95.0%), while fragments comprised 4.4%, and
films and foams comprising less than 1% MP combined. Seventy percent (n = 981) of
C. virginica contained MP in their tissues. When considered by region, 65.5% of north
samples, 70.4% of central samples, and 76.7% of south samples contained MP. The dominant
color of MPs lagoon-wide and for each region was black. Plastics ranged in size from 0.1
to 35 mm, with a mean size (±CI) of 2.79 ± 0.10 mm. Oysters had a mean MP abundance
(±CI) of 2.26 ± 0.16 MP/individual and density of 2.43 ± 0.52 MP/g wet tissue weight.
MP abundance differed between C. virginica from IRL regions; northern oysters contained
less MP than central and south oysters (p < 0.001, Figure 3), but abundance did not differ
between the central and south oysters.
 of central samples, and 76.7% of south samples contained MP. The dominant color of MPs
lagoon-wide and for each region was black. Plastics ranged in size from 0.1 to 35 mm, with
a mean size (±CI) of 2.79 ± 0.10 mm. Oysters had a mean MP abundance (±CI) of 2.26 ±
0.16 MP/individual and density of 2.43 ± 0.52 MP/g wet tissue weight. MP abundance dif-
fered between C. virginica from IRL regions; northern oysters contained less MP than cen-
Environments 2022, 9, 131
tral and south oysters (p < 0.001, Figure 3), but abundance did not differ between the9cen-
of 18

tral and south oysters.

Figure 3.
Figure Microplastic abundance
3. Microplastic abundance by
by region
region in
in Indian
Indian River
River Lagoon
Lagoon oysters.
oysters. Values
Values reported
reported are
are the
the
mean
mean (point)
(point) and
and the
the 95%
95% confidence
confidence interval
interval ofof the mean (error bar). (GLM,
(GLM, p == 0.01,
0.01, North
North ==710,
710,
Central
Central ==345,
345,South
South ==347).
347).

The
The most plausible model model to to predict
predictMP MPabundance
abundancethroughout
throughoutthe theIRLIRLforforC.C.virginica
virgin-
ica included
included season,
season, site,
site, andand shellheight
shell heightasaspredictor
predictorvariables
variables(Table
(Table3).3). MP abundance
abundance
was higher in in oysters
oysters sampled
sampled in in summer,
summer, fall, and winter winter (p(p <<0.03
0.03for
forall)
all)than
thanin inspring.
spring.
Within each region, there was variability in MP abundances (p < 0.05). In In north
north IRL,
IRL, the
the
most plausible model included site + season (AIC = 2376.0; McFadden pseudoR 2 = 0.11,
most plausible model included + season = McFadden pseudoR = 0.11, 2

Table 3),
3),while
whileininthethecentral
centralandand south
south IRL, IRL, season
season + shell
+ shell heightheight produced
produced the mostthe plau-
most
plausible
sible models models
(Table (Table 3). Abundance
3). Abundance was higher
was higher in central
in central C. virginica
C. virginica in summerin summer
and fall and
(p
<fall (p < 0.001).
0.001). The model The indicated
model indicated MP abundance
MP abundance increased increased by 1.4 MP/individual
by 1.4 MP/individual for everyfor 1
everyincrease
mm 1 mm increase
in shellin shell height
height (p < 0.001)(p < 0.001)
in the in the north.
north. The same
The same modelmodel indicated
indicated that that
MP
MP abundance
abundance increased
increased by 1.51
by 1.51 MP/individual
MP/individual for every
for every 1 mm1increase
mm increase
in shellin height
shell height
in C.
in C. virginica from the central IRL, and abundance increased
virginica from the central IRL, and abundance increased by 1.45 MP for every 1 mm by 1.45 MP for every 1 mm
in-
increase
crease in in shell
shell height
height ≤ 0.002)
(p ≤(p0.002) in the
in the south.south.
When When the presence/absence
the presence/absence of MPs of was
MPscon-was
considered
sidered by region,
by region, summer,
summer, fall,fall,
andand winter
winter were were significant
significant predictors
predictors of MP
of MP presence
presence in
in oysters
oysters fromfromthethe northern
northern region
region (p(p≤ ≤ 0.04).
0.04). Similarly,inincentral
Similarly, centralIRL,
IRL,summer,
summer, fall,fall, and
and
winter were
winter were all all predictors
predictors of of MPMP presence
presence (p (p ≤ 0.04) for
≤ 0.04) C. virginica.
for C. virginica. No No variable
variable was was aa
significant predictor of MP presence for
significant predictor of MP presence for south oysters. south oysters.
Microplastic abundance trends in C. virginica differed between IRL regions with regard
to oceanic and freshwater influence. In the northern and central IRL, MP abundance in
oysters decreased with every 1 km increase in distance from a freshwater tributary, by
0.77 MP/individual and 0.52 MP/individual, respectively (p ≤ 0.01 for both). However,
no oceanic influence on MP abundance was detected in any region, and in the southern
IRL, there was also no tributary influence.

3.3. Abiotic Parameters

Air and water temperature differed significantly between IRL regions and seasons
(p < 0.005 for all), but did not differ between sampling sites for water and oyster collections.
As expected, temperatures were higher in summer and spring, and lower in winter. The
water temperature was also higher in the southern region. Salinity did not vary between
regions or seasons but did vary between sites (p < 0.05 for all). Wind speed varied between
regions and seasons, but not months. Wind speed was slower in the central IRL region
(p = 0.004), and faster at northern water sampling sites that were more exposed (p < 0.01).
Environments 2022, 9, 131 10 of 18

Table 3. Zero-inflated negative binomial GLM models of MP abundance in Indian River Lagoon oys-
ters, overall and by region. Values reported are AIC, delta AIC, degrees of freedom, and AIC weight.

Indian River Lagoon Oysters AIC ∆AIC df AIC Weight

Site + season + shell height 4984.0 0.0 35 1
Season + shell height 5020.6 32.6 13 <0.001
Site + shell height 5053.1 69.1 29 <0.001
Shell height 5088.2 104.2 7 <0.001
Site + season 5158.6 174.6 35 <0.001
Season 5208.4 224.4 13 <0.001
Site 5248.5 264.5 29 <0.001
Tributary 5277.0 293.0 9 <0.001
Region 5289.1 305.1 11 <0.001
North Lagoon AIC ∆AIC df AIC Weight
Site + season 2376.0 0.0 23 0.63
Site + season + shell height 2377.1 1.0 23 0.37
Season + shell height 2404.2 28.2 13 <0.001
Season 2407.2 31.2 13 <0.001
Site 2411.0 34.9 17 <0.001
Site + shell height 2413.5 36.5 17 <0.001
Tributary 2435.6 59.6 9 <0.001
Inlet 2435.6 59.6 9 <0.001
Shell height 2436.5 60.5 7 <0.001
Central Lagoon AIC ∆AIC df AIC Weight
Season + shell height 1203.5 0.0 13 1
Shell height 1263.2 59.7 7 <0.001
Site + season 1289.7 86.2 15 <0.001
Season 1310.2 106.7 13 <0.001
Site 1364.5 161.0 9 <0.001
Tributary 1371.7 168.2 9 <0.001
South Lagoon AIC ∆AIC df AIC Weight
Season + shell height 1335.8 0.0 13 1
Shell height 1372.0 36.2 7 <0.001
Season 1408.7 72.9 13 <0.001

3.4. Polymer Composition and Contamination

In total, 122 signals of suspected MPs were obtained using ATR-FTIR spectroscopy,
and 78 (64%) were confirmed synthetic polymers. Fibers, fragments, foams, and films
were found in both lagoon water and C. virginica. Fibers dominated type composition,
comprising 95.6% and 95.0% MPs in water and C. virginica, respectively. Colors varied
across the spectrum, but black MPs were the most common. It is possible that the citizen
scientists who collected field samples may have contaminated their samples if their clothing
was not created from natural materials [49,50]. We acknowledge this potential source of
error. Project scientists ensured that all individuals involved in laboratory MP microscopy
wore only natural fiber garments.
Polyethylene terephthalate (PET) was the most abundant polymer in lagoon water
and C. virginica in the IRL, and comprised 50%, and 56% MPs, respectively. Polypropy-
lene (PP), polyethylene (PE), polystyrene (PS), and polyamide (PA) were also found in
lagoon water in differing proportions (Figure 4). All scanned MPs were fibers, except for
two clear fragments. One fragment was PE and the other was a synthetic wax. There was
one rayon fiber found and it was in an oyster. Miscellaneous (“other”) polymers found
in both water and oyster samples were polymer blends, with the exception of one acrylic
adhesive confirmed in lagoon water, and one polyacrylate fiber confirmed in C. virginica.
 (PP), polyethylene (PE), polystyrene (PS), and polyamide (PA) were also found in lagoon
water in differing proportions (Figure 4). All scanned MPs were fibers, except for two
clear fragments. One fragment was PE and the other was a synthetic wax. There was one
rayon fiber found and it was in an oyster. Miscellaneous (“other”) polymers found in both
water and oyster samples were polymer blends, with the exception of one acrylic adhesive
Environments 2022, 9, 131 11 of 18
confirmed in lagoon water, and one polyacrylate fiber confirmed in C. virginica.

Water Oysters
3%
6%
6% PET PET
9% PP 3% 22% PP
PE PA
50%
PA 56% PS
26% 13%
PS 3% Rayon
Other Other
3%

Figure 4. Synthetic
Figure 4. Synthetic polymers polymersfrom
in microplastics in microplastics
water and from water
oysters andthe
from oysters from
Indian the Indian
River Lagoon.River Lagoon.
Polymersterephthalate
Polymers are polyethylene are polyethylene terephthalate
(PET), (PET), polypropylene
polypropylene (PP), polyethylene
(PP), polyethylene (PE), polyamide
(PE), polyamide
(PA),rayon,
(PA), polystyrene (PS), polystyrene (PS), rayon, andpolymers
and miscellaneous miscellaneous polymers (Other).
(Other).

Aerial control blanks associated with lagoon water had a mean contamination rate of
Aerial control blanks associated with lagoon water had a mean contamination rate of
0.016 MP/min. The mean time of exposure associated with microscopy for water samples
0.016 MP/min. The mean time of exposure associated with microscopy for water samples
was 8 min. Mean aerial contamination rate for C. virginica samples was 0.020 MP/min
was 8 min. Mean aerial contamination
and samples rate for
were exposed foraC. virginica
mean samples
time of was 0.020
5 min. There was aMP/min and(three PET
15% overlap
samples were exposed forbetween
particles) a meanpolymers
time of found
5 min.inThere
sampleswas
andapolymers
15% overlap (threecontrol
from aerial PET blanks.
particles) between polymers found in samples and polymers from aerial control blanks.
4. Discussion
4. Discussion A growing body of research, including our results, suggest oceans function as sinks
for MPs, while coastal surface waters entering estuaries are sources [51–53]. The current
A growing body of research, including our results, suggest oceans function as sinks
study quantified MP abundance in surface water and C. virginica from the IRL to determine
for MPs, while coastal surface
if spatial waters entering
and temporal estuariesMP
factors influence are abundances
sources [51–53].
withinThe
thiscurrent
system. Overall,
study quantified MP abundance in surface water and C. virginica from the
C. virginica had an average of 2.26 MP/individual, or 2.43 MP/g tissue IRL to deter-
weight, and IRL
mine if spatial andwater
temporal factors
had 1.47 MP/L. influence MPvariations
Significant abundances andwithin
trends this system.
in MP Overall,
abundance were detected
C. virginica had an average
across of 2.26
seasons, and MP/individual, or 2.43
within spatial extents less MP/g
than 5 tissue weight,both
km, indicating and site
IRLand season
should
water had 1.47 MP/L. be incorporated
Significant into MP
variations andresearch
trendsdesigns. For example,were
in MP abundance our research
detecteddetermined
across seasons, andthat the south
within IRL was
spatial a hotspot
extents for MP
less than pollution.
5 km, indicating both site and season
Studies of estuarine and coastal bivalves have reported variable MP abundances
(e.g., [27,54,55]). Comparisons of MP abundance in Florida’s IRL to abundances of other
estuaries in the United States are summarized in Table 4. Compared to a previous study on
MPs in the north IRL by [56], our abundance values for both lagoon water and adult oysters
were lower even though the oyster reefs were in close proximity (separated by < 1 km). This
difference can likely be attributed to the different MP collection and extraction procedures
between studies, as FTIR was not included in [56], and we now know that large numbers
of natural fibers are present in the IRL system (C.A.C., pers. obs.). Additionally, aerial
contamination was not reported in the Waite et al. [56] publication. MP abundance in IRL
water was comparable to mean abundances in a surface water study from Tampa Bay (FL),
published in 2019, but not to earlier studies from highly urbanized Charleston Harbor, SC
or less urbanized Winyah Bay, SC [55,57]. It is important to note that the south IRL, where
mean MP abundance exceeded 2 MP/L, is the most highly urbanized area within the IRL
system and some of the sampling locations were along human-built canals. When oysters
were compared, MP abundance in C. virginica from the IRL was less than MP abundance
in C. gigas from the Oregon coast, but was similar to C. gigas in the Salish Sea, WA [58,59].
This may be related to oyster filtration rates, month or time of sampling, MP protocols,
Environments 2022, 9, 131 12 of 18

or localized urbanization. Filtration rates for C. gigas from the Pacific Ocean have been
measured in the field at 0.35–0.73 L g−1 h−1 and 2.5–12 L g−1 h−1 in laboratory trials (dry
tissue weight; [60–64]). There are reports that this species can filter in “high gear” vs. “low
gear”, with a 3-fold difference in filtration [65]. Galimany et al. [66] reported that adult
IRL C. virginica filter 18 L day−1 (0.75 L h−1 ) in flow-through chambers using lagoon water,
while Grizzle et al. [67] reported 1.2 L h−1 filtration rates in shallow IRL field conditions.
The MP abundances reported in this study were also comparable to those from C. gigas
collected along the French Atlantic coast (1.7 MP/individual; [68]).

Table 4. Comparison of microplastic abundance in water and oysters from the Indian River Lagoon
and other estuaries in the United States. Values reported are mean abundance per liter of water,
microplastics per individual oyster, and standard error of the mean.

Water Location Abundance ± S.E. Reference

Indian River Lagoon, FL 1.46 ± 0.05 Present study
Mosquito Lagoon, FL 23.1 [56]
Tampa Bay Estuary, FL 0.94 ± 0.52 [55]
Charleston Harbor, SC 6.6 ± 1.3 [57]
Winyah Bay, SC 30.8 ± 12.1 [57]
Oysters Location Abundance ± S.E. Reference Species
Indian River Lagoon, FL 2.26 ± 0.08 Present study Crassostrea virginica
Mosquito Lagoon, FL 16.5 [56] Crassostrea virginica
Salish Sea, WA 1.75 [59] Crassostrea gigas
Oregon Coast 10.95 ± 0.77 [58] Crassostrea gigas

4.1. Spatial Microplastic Fluctuations and Influences

MP abundance in both lagoon water and C. virginica were influenced differently by
hydrological factors of distance to a tributary or inlet. Freshwater contribution to the IRL
comes from land runoff and a dynamic matrix of rivers, drainage canals, creeks, and ditches,
which are unevenly distributed throughout the lagoon [29]. In the northern IRL, there
was no tributary or inlet influence detected on MP abundance in water or oysters. There
are no tributaries and the north IRL is considered microtidal with water residence times
(50% renewal time) within the region varying greatly with distance from inlet; for example,
residence time for the northernmost and southernmost portions of Mosquito Lagoon (north
IRL) are 15 days and 172 days, respectively [30]. Within the northern IRL, MP abundance
in water did not differ between sites; however, abundance was different in oysters from
different reefs. All northern sampling reefs were located within 20 km of Ponce de Leon
Inlet and within 5 km of each other so spatial influences on MP abundances in C. virginica
were hard to distinguish. All reefs were located in the central Mosquito Lagoon region,
where water residence time is low (~15 days; [30]). Likewise, abiotic conditions between
sampled reefs in the north IRL did not differ. It is likely that the variation in MP abundance
in the north IRL is not explained by variables captured in this study, and additional research
is needed. One possibility is that the location of stormwater outfalls, not incorporated in
this study, are important sources of MP pollution in this region of the IRL (L.J.W., pers.
comm.; [3,69]).
In the central IRL, a positive trend of freshwater influence on MP abundance was
found in both water and C. virginica, as abundance decreased with increasing distance from
a tributary. Tributaries within the boundaries of the central IRL are the Sebastian and Eau
Gallie Rivers, Turkey and Crane Creeks, and numerous manmade canals (e.g., Vero Main,
Vero North, Vero South, Taylor; [29]). The Sebastian River is the second largest tributary into
the IRL [70]. Despite the greater tributary presence, water from the central IRL contained
the lowest MP abundances, which may be attributed to the inlets flushing MPs out of the
region. There are two inlets within the boundaries of the central IRL, Sebastian and Fort
Pierce, which contribute to lower water residence times and increased tidal flushing in the
Environments 2022, 9, 131 13 of 18

region [71]. In oysters from the central IRL, however, there was no inlet influence while
MP abundance increased with closer proximity to freshwater tributaries. This may be
attributed to the proximity of two of the three reefs to tributaries; both were adjacent to
tributaries that empty into the lagoon, while the third reef was 3 km away from Fort Pierce
Inlet. Additionally, this third reef was located on the western side of the central IRL, where
there is a lessened tidal influence compared to the eastern shore [72]. Another important
factor for MP abundance in water and oysters in the central IRL was discharge from the
Sebastian River, which had a mean annual discharge rate of ~100 m3 s−1 throughout this
study [72]. Law et al. [73] found that MPs accumulate in areas with water velocities slower
than 2 cm s−1 , suggesting MPs in this area are flushed away by the water velocity of the
Sebastian River.
Southern IRL MP abundance was high for both C. virginica and lagoon water. MP
abundance in southern lagoon water decreased with increasing distance from a tributary;
however, MP abundance in C. virginica did not. There is one primary tributary in the
south IRL, the St. Lucie Estuary (SLE), which is also the largest tributary to the IRL and
connects the lagoon to Lake Okeechobee through the C44 canal [29]. MP abundance was
not influenced by distance to an inlet, in either lagoon water or C. virginica. There is one
inlet within the southern IRL boundary defined in this study, the St. Lucie Inlet, which has
constricted water flow into the area [74]. As a result, there is less tidal influence in this area
and in the SLE [74]. This suggests the St. Lucie Inlet is not flushing MP out of the southern
IRL at rates fast enough to accommodate deposition from the tributary. Abundance in
C. virginica in the southern IRL did not differ between sampling reefs. Similar abundances
in southern reefs may be attributed to their location, as they were all north of the SLE and
St. Lucie Inlet (>5 km) where water circulation patterns are less impacted by either the
tributary or inlet [49,74].

4.2. Temporal Microplastic Fluctuations and Influences

Our sampling design included one collection per month (IRL water) or per season
(oysters), so it is essential to consider this limitation to the study when examining tem-
poral trends in MPs in the IRL, especially with oysters. With IRL water overall and for
all three regions, MP abundance was higher in the fall and winter during our study. Tem-
poral variation in MP abundance was likely impacted by extreme events that happened
during the study period. Hurricane Dorian, which paralleled Florida’s east coast between
1 and 3 September 2019, may have impacted MP abundance in the lagoon. Hitchcock [25]
found MP abundance levels in the Cooks River Estuary (Australia) were 40-fold higher
during a storm event. Around the time of Hurricane Dorian, discharge out of the Sebastian
River and SLE increased to ~305 m3 s−1 [73]. There were also prolonged high-water levels
associated with a new lunar cycle and slowing of the Atlantic current in November 2019,
which may also have influenced water discharges, and thus, MP abundance [73].
With IRL oysters, the overall pattern suggests that spring had lower MP abundances
than other seasons. This pattern may be related to seasonal oyster eco-physiology and
seasonal differences in their pumping rates. Pumping rates increase with temperature [75].
Likewise, oyster reproduction increases with temperature and Florida oysters begin to
spawn each year when temperatures reach or exceed 25 ◦ C [40]. In the north IRL, C. virginica
recruited to reefs in all months of the year except January–March throughout a 6-year study;
≥5 spat were recorded per 0.25 m2 as long as the water temperature from nearby continuous
monitoring was ≥19.3 ◦ C [76]. Spring oysters were collected in March 2019 when water
temperatures were below this temperature threshold. Combined, this suggests that lowered
pumping and reproductive activity by oysters may result in lower MP abundance. This
could have implications for harvesting and human consumption, but more detailed studies
on this topic are required. Craig et al. [21] found that C. virginica, regardless of size, were
able to egest MPs at a mean rate of 1 MP per 1 h through feces, and 1 MP per 2 h through
pseudofeces, and that egestion efficiency decreased by 0.8% per 1 g increase in tissue weight.
Environments 2022, 9, 131 14 of 18

This corresponds to the current finding that size is important as MP abundance increased
by 1.4–1.5 MP/individual for every 1 mm increase in shell height.

4.3. Polymers and Coastal Restoration

Development and field testing of non-plastic materials have become important top-
ics in the field of coastal restoration to reduce any unintended consequences of plastic
restoration materials on the environment [77]. Negative assumptions continue to be made
about plastic-based restoration materials contributing to overall MP abundances without
supporting data. Materials traditionally used for restoration borrowed heavily from the
aquaculture industry and many areas around the globe use black, Naltex (PE) mesh bags
for oyster reef restoration and living shoreline stabilization (e.g., [78,79]). The mesh bags
are filled with shells, often from shell recycling programs, and deployed as the bases for
reefs or as wave breaks along shorelines. This made sense as this bagging mesh was
non-toxic, volunteer-friendly, and low cost. However, while aquaculturists may leave
materials in water bodies for finite time frames, this was not the plan for many restoration
projects. Hence, there is a community concern that these PE bags deployed for restoration
are now adding to the MP problem in the IRL and elsewhere. We were able to directly
test if C. virginica collected from the south IRL had a large number of black PE fibers or
fragments as all three study reefs were previously restored using Naltex bags (V.E., pers.
Comm.). Although black was the dominant color of fibers that dominated both water
and oyster MP in south IRL, it is unlikely that the source was the Naltex mesh bags as PE
signatures were limited (9% in water, < 2% in oysters). However, the minimum sample size
for FTIR was 0.5 mm; thus, the number of Naltex mesh bag MP of smaller dimensions is
not known. Craig et al. [21] likewise found very few (<5%) PE MPs in oyster biodeposits
(feces, pseudofeces) and 8% in the soft tissues of 280 individuals of C. virginica collected
throughout the IRL in a separate study.
There are many alternative sources of MPs in the IRL and around the globe. Polyethy-
lene terephthalate (PET) was the dominant polymer found in both C. virginica and water
in our study and the IRL oyster biodeposits study by Craig et al. [20]. PET was the most
abundant polymer in feces, pseudofeces, and oyster tissue, and comprised 80%, 50%, and
58% confirmed MP, respectively [21]. PET is prominent in the single-use plastic industry,
particularly plastic drink bottles [80–82]. Polyester, also known as PET, is the most pro-
duced synthetic textile material in the world and is common in clothing [83]. Since 95%
of MPs in the IRL were fibers, it is possible that some MPs originated from wastewater
treatment plants or septic systems. Another source is stormwater outfalls, and IRL outfall
locations have higher MP abundances than other regions (Walters et al., unpublished data).
Globally, Boucher and Friot [7] estimate 35% of plastics in oceans are from synthetic textiles
associated with laundry.
Of the 44 misidentified non-polymers, 39% were natural textile fibers, including wool,
cotton, and silk fibroin [84]. An additional 30% were cellulose derivatives (e.g., microcrys-
talline, microfibrillated cellulose), and 10% were ramie fiber. These are fibers engineered to
be resistant to breakage, suggesting that there may be a weakness in identification proce-
dures where resistance to breakage may be too heavily relied upon as a characteristic to
classify a particle as an MP.

5. Conclusions
Over the period of this study, 84 trained citizens participated in water sampling,
processing, and inspecting, and contributed 1600 h of their time to Indian River Lagoon
MP research. In the IRL, MP abundance was variable, both spatially and temporally, which
can be attributed to the unique hydrology of this 251 km long estuary along the east coast
of central Florida. In total, 6936 MPs were found in the IRL (water + oysters) collections,
95% of which were fibers, and the majority was PET. The southern IRL was a hotspot for
microplastic pollution; this region is highly urbanized and also includes the largest tributary
to the IRL. Overall, freshwater tributaries in the central and south IRL were the suggested
Environments 2022, 9, 131 15 of 18

sources of MP pollution, while the Sebastian and Ft. Pierce inlets flushed MPs out of this
system. Using a mean abundance of 1.5 MP/L and lagoon volume of 953,000,000 cm3 [85],
we estimate there are ~1.4 trillion MPs in the Indian River Lagoon. This research builds
on previous IRL MP research as well as the rapidly expanding global body of research
of spatial and temporal variations in MP abundance in dynamic estuarine systems that
include both highly urbanized and undeveloped areas.

Author Contributions: Conceptualization, L.J.W., E.D., J.W. and V.E.; methodology, L.J.W., C.A.C.,
E.D. and J.W.; validation, L.J.W. and C.A.C.; formal analysis, C.A.C.; investigation, L.J.W., C.A.C., E.D.,
T.S.-T. and G.C.; resources, L.J.W., E.D., J.W., V.E. and L.Z.; data curation, C.A.C.; writing—original
draft preparation, C.A.C.; writing—review and editing, L.J.W., E.D., J.W., V.E., T.S.-T., G.C., D.W.F.
and L.Z.; visualization, C.A.C.; supervision, L.J.W., and L.Z; project administration, L.J.W.; funding
acquisition, L.J.W., E.D., J.W. and V.E. All authors have read and agreed to the published version of
the manuscript.
Funding: This research was funded by the Indian River Lagoon National Estuary Program and
National Science Foundation award #1617374.
Data Availability Statement: Data are available on UCF STARS digital repository at https://stars.
library.ucf.edu (accessed on 24 August 2022).
Acknowledgments: We thank all the citizen-scientists, staff and research assistants from University of
Central Florida, Florida Oceanographic Society, Marine Discovery Center, Indian River Lagoon Aquatic
Preserve, Oxbow Eco-Center, and Environmental Learning Center who dedicated their time, space and
equipment to complete this project. In particular, we thank K. Fusco, P. Sacks, A. Wright, W. Giles, S.
Busch, M. Keplinger, S. Krulis and J. Copertino for field and laboratory assistance. Finally, we thank the
National Park Service (Canaveral National Seashore) for providing access to north IRL sites.
Conflicts of Interest: The authors declare no conflict of interest. The funders had no role in the design
of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or
in the decision to publish the results.

References
1. Sekudewicz, I.; Dabrowska, A.M.; Syczewski, M.D. Microplastic Pollution in Surface Water and Sediments in the Urban Section
of the Vistula River (Poland). Sci. Total Environ. 2021, 762, 143111. [CrossRef] [PubMed]
2. Zhao, J.; Ran, W.; Teng, J.; Liu, Y.; Liu, H.; Yin, X.; Cao, R.; Wang, Q. Microplastic Pollution in Sediments from the Bohai Sea and
the Yellow Sea, China. Sci. Total Environ. 2018, 640, 637–645. [CrossRef] [PubMed]
3. Kalavrouziotis, I.; Karapanagioti, H. Microplastics in Water and Wastewater, 2nd ed.; IWA Publishing: London, UK, 2020.
4. Baekeland, L.H. The Synthesis, Constitution, and Uses of Bakelite. Ind. Eng. Chem. 1909, 1, 149–161. [CrossRef]
5. Crespy, D.; Bozonnet, M.; Meier, M. 100 years of Bakelite, the Material of 1000 uses. ChemInform 2008, 39, 3322–3328. [CrossRef]
6. Geyer, R.; Jambeck, J.R.; Law, K.L. Production, Use, and Fate of All Plastics Ever Made. Sci. Adv. 2017, 3, e1700782. [CrossRef]
7. Boucher, J.; Friot, D. Primary Microplastics in the Oceans: A Global Evaluation of Sources; IUCN: Gland, Switzerland, 2017; pp. 227–229.
8. Jambeck, J.R.; Geyer, R.; Wilcox, C.; Siegler, T.R.; Andrady, A.; Narayan, R.; Law, K.L. Plastic Waste Inputs from Land into the
Ocean. Science 2015, 347, 768–771. [CrossRef]
9. Arthur, C.; Baker, J.; Bamford, H. (Eds.) Proceedings of the International Research Workshop on the Occurrence, Effects and Fate
of Microplastic Marine Debris. In NOAA Technical Memorandum NOS-OR&R-30; University of Washington Tacoma: Tacoma,
Washington, DC, USA, 2009.
10. Guo, X.; Wang, J. The Chemical Behaviors of Microplastics in Marine Environment: A Review. Mar. Pollut. Bull. 2019, 142, 1–14.
[CrossRef]
11. Barnes, D.K.; Galgani, F.; Thompson, R.C.; Barlaz, M. Accumulation and Fragmentation of Plastic Debris in Global Environments.
Phil. Trans. R. Soc. B Biol. Sci. 2009, 364, 1985–1998. [CrossRef]
12. Auta, H.S.; Emenike, C.U.; Fauziah, S.H. Distribution and Importance of Microplastics in the Marine Environment: A Review of
the Sources, Fate, Effects, and Potential Solutions. Environ. Int. 2017, 102, 165–176. [CrossRef]
13. Ellison, K. The Trouble with Nurdles. Front. Ecol. Environ. 2007, 5, 396. [CrossRef]
14. Li, H.X.; Ma, L.S.; Lin, L.; Ni, Z.X.; Xu, X.R.; Shi, H.H.; Yan, Y.; Zheng, G.; Rittschof, D. Microplastics in Oyster Saccostrea cucullata
Along the Pearl River Estuary, China. Environ. Pollut. 2018, 236, 619–625. [CrossRef]
15. Simon-Sanchez, L.; Grelaud, M.; Garcia-Orellana, J.; Ziveri, P. River Deltas as Hotspots of Microplastic Accumulation: The Case
Study of the Ebro River (NW Mediterranean). Sci. Total Environ. 2019, 687, 1186–1196. [CrossRef]
16. Luo, W.; Su, L.; Craig, N.J.; Du, F.; Wu, C.; Shi, H. Comparison of Microplastic Pollution in Different Water Bodies from Urban
Creeks to Coastal Waters. Environ. Pollut. 2019, 246, 174–182. [CrossRef]
Environments 2022, 9, 131 16 of 18

17. Wang, F.J.; Tan, Z.; Peng, J.; Qiu, Q.; Li, M. The Behaviors of Microplastics in the Marine Environment. Mar. Environ. Res. 2016,
113, 7–17. [CrossRef]
18. Zhang, F.; Man, Y.B.; Mo, W.Y.; Man, K.Y.; Wong, M.H. Direct and Indirect Effects of Microplastics on Bivalves, with a Focus on
Edible Species: A Mini-Review. Crit. Rev. Environ. Sci. Technol. 2020, 50, 2109–2143. [CrossRef]
19. Cho, Y.; Shim, W.J.; Jang, M.; Han, G.M.; Hong, S.H. Nationwide Monitoring of Microplastics in Bivalves from the Coastal
Environment of Korea. Environ. Pollut. 2021, 270, 116175. [CrossRef]
20. Ward, J.E.; Zhao, S.; Holohan, B.A.; Mladinich, K.M.; Griffin, T.W.; Wozniak, J.; Shumway, S.E. Selective Ingestion and Egestion of
Plastic Particles by the Blue Mussel (Mytilus edulis) and Eastern Oyster (Crassostrea virginica): Implications for Using Bivalves as
Bioindicators of Microplastic Pollution. Environ. Sci. Technol. 2019, 53, 8776–8784. [CrossRef]
21. Craig, C.A.; Fox, D.W.; Zhai, L.; Walters, L.J. In-Situ Microplastic Egestion Efficiency of the Eastern Oyster Crassostrea virginica.
Mar. Pollut. Bull. 2022, 178, 113653. [CrossRef]
22. Eierman, L. Multiple Negative Impacts of Polyethylene Terephthalate Plastic (PET) Exposure on Juvenile Eastern Oysters
(Crassostrea virginica). In Proceedings of the Coastal & Estuarine Research Federation 25th Biennial Conference, Mobile, AL, USA,
3–7 November 2019.
23. Newell, R.I.; Jordan, S.J. Preferential Ingestion of Organic Material by the American Oyster Crassostrea virginica. Mar. Ecol. Prog.
Ser. 1983, 13, 47–53. [CrossRef]
24. Sussarellu, R.; Suquet, M.; Thomas, Y.; Lambert, C.; Fabioux, C.; Pernet, M.; Le Goic, N.; Quillien, V.; Mignant, C.; Epelboin, Y.; et al.
Oyster Reproduction is Affected by Exposure to Polystyrene Microplastics. Proc. Natl. Acad. Sci. USA 2016, 113, 2430–2435.
[CrossRef]
25. Hitchcock, J.N. Storm Events as Key Moments of Microplastic Contamination in Aquatic Ecosystems. Sci. Total Environ. 2020,
734, 139436. [CrossRef]
26. Xia, W.; Rao, Q.; Deng, X.; Chen, J.; Xie, P. Rainfall is a Significant Environmental Factor of Microplastic Pollution in Inland Waters.
Sci. Total Environ. 2020, 732, 139065. [CrossRef]
27. Lima, A.R.A.; Barletta, M.; Costa, M.F. Seasonal Distribution and Interactions between Plankton and Microplastics in a Tropical
Estuary. Estuar. Coast. Shelf Sci. 2015, 165, 213–225. [CrossRef]
28. Zhang, Y.L.; Wang, F.X.; Shock, C.C.; Yang, K.J.; Kang, S.Z.; Qin, J.T.; Li, S.E. Effects of Plastic Mulch on the Radiative and Thermal
Conditions and Potato Growth under Drip Irrigation in Arid Northwest China. Soil Tillage Res. 2017, 172, 1–11. [CrossRef]
29. Indian River Lagoon National Estuary Program. 2030 Comprehensive Conservation and Management Plan. 2019. Available
online: https://onelagoon.org/management-plan/ (accessed on 19 August 2022).
30. Rosario-Llantin, J.A.; Zarillo, G.A. Flushing Rates and Hydrodynamical Characteristics of Mosquito Lagoon (Florida, USA).
Environ. Sci. Pollut. Res. 2021, 28, 30019–30034. [CrossRef]
31. Smith, N.P. Tidal and Nontidal Flushing of Florida’s Indian River Lagoon. Estuaries 1993, 16, 739–746. [CrossRef]
32. Walters, L.; Sacks, P.; Campbell, D. Boating Impacts and Boat-Wake Resilient Restoration of the Eastern Oyster Crassostrea virginica
in Mosquito Lagoon, Florida, USA. Fla. Sci. 2021, 84, 173–199.
33. Lewis, D.; Durham, K.; Walters, L.; Cook, G. Resident Fishes as Higher Trophic Level Indicators of Oyster Reef Restoration
Success. Sustainability 2021, 13, 13004. [CrossRef]
34. Loch, J.M.H.; Walters, L.J.; Donnelly, M.L.; Cook, G.S. Restored Coastal Habitat Can “Reel In” Juvenile Sportfish: Population and
Community Responses in the Indian River Lagoon, Florida, USA. Sustainability 2021, 13, 12832. [CrossRef]
35. Copertino, J.; Harris, K.; Chute, L.; Walters, L. Impact of Oyster (Crassostrea virginica) Reef Restoration on Benthic Invertebrates
and Coastal Birds in a Subtropical Estuary. Sustainability 2022, 14, 2371. [CrossRef]
36. Barber, A.; Walters, L.; Birch, A. Potential for Restoring Biodiversity of Macroflora and Macrofauna on Oyster Reefs in Mosquito
Lagoon, Florida. Fla. Sci. 2010, 73, 47–62.
37. Searles, A.; Gipson, E.; Walters, L.; Cook, G. Oyster Reef Restoration Facilitates the Recovery of Marine Invertebrate Abundance,
Diversity, and Composition in Estuarine Communities. Nat. Sci. Rep. 2022, 12, 8163. [CrossRef] [PubMed]
38. Cutroneo, L.; Reboa, A.; Besio, G.; Borgogno, F.; Canesi, L.; Canuto, S.; Dara, M.; Enrile, F.; Forioso, I.; Greco, G.; et al. Microplastics
in Seawater: Sampling Strategies, Laboratory Methodologies, and Identification Techniques Applied to Port Environment. Environ.
Sci. Pollut. Res. 2020, 27, 8938–8952. [CrossRef] [PubMed]
39. Zhu, J.; Zhang, Q.; Li, Y.; Tan, S.; Kang, Z.; Yu, X.; Lan, W.; Cai, L.; Wang, J.; Shi, H. Microplastic Pollution in the Maowei Sea, a
Typical Mariculture Bay of China. Sci. Total Environ. 2019, 658, 62–68. [CrossRef]
40. Radabaugh, K.R.; Geiger, S.P.; Moyer, R.P. (Eds.) Oyster Integrated Mapping and Monitoring Program Report for the State of Florida;
Technical Report 22; Florida Fish and Wildlife Conservation Commission, Fish and Wildlife Research Institute: St. Petersburg, FL,
USA, 2019.
41. Garvis, S.K.; Sacks, P.E.; Walters, L.J. Formation, Movement, and Restoration of Dead Intertidal Oyster Reefs in Canaveral
National Seashore and Mosquito Lagoon, Florida. J. Shellfish Res. 2015, 34, 251–258. [CrossRef]
42. Shaw Institute. Guide to Microplastics Identification, A Comprehensive Methods Guide for Microplastics Identification and
Quantification in the Laboratory. Shaw Institute: Blue Hill, ME, USA, 2019. Available online: https://www.shawinstitute.org/
(accessed on 1 September 2019).
43. Thiele, C.J.; Hudson, M.D.; Russell, A.E. Evaluation of Existing Methods to Extract Microplastics from Bivalve Tissue: Adapted
KOH Digestion Protocol Improves Filtration at Single-Digit Pore Size. Mar. Pollut. Bull. 2019, 142, 384–393. [CrossRef]
Environments 2022, 9, 131 17 of 18

44. Foekema, E.M.; de Gruijter, C.; Mergia, M.T.; van Franeker, J.A.; Murk, A.J.; Koelmans, A.A. Plastic in North Sea Fish. Environ.
Sci. Technol. 2013, 47, 8818–8824. [CrossRef]
45. Granek, E.F.; Brander, S.; Holland, E.B. Microplastics in Aquatic Organisms: Improving Understanding and Identifying Directions
for the Next Decade. Limnol. Oceanogr. Lett. 2020, 5, 1–4. [CrossRef]
46. Gago, J.; Galgani, F.; Maes, T.; Thompson, R.C. Microplastics in Seawater: Recommendations from the Marine Strategy Framework
Directive Implementation Process. Front. Mar. Sci. 2016, 3, 219. [CrossRef]
47. Frias, J.P.G.L.; Gago, J.; Otero, V.; Sobral, P. Microplastics in Coastal Sediments from Southern Portuguese Shelf Waters. Mar.
Environ. Res. 2016, 114, 24–30. [CrossRef]
48. R Core Team. R: A Language and Environment for Statistical Computing; R Foundation for Statistical Computing: Vienna, Austria,
2019; Available online: https://www.R-project.org/ (accessed on 1 December 2018).
49. Gwinnett, C.; Miller, R.Z. Are We Contaminating Our Samples? A Preliminary Study to Investigate Procedural Contamination
During Field Sampling and Processing for Microplastic and Anthropogenic Microparticles. Mar. Pollut. Bull. 2021, 173, 113095.
[CrossRef]
50. Rota, E.; Bergami, E.; Corsi, I.; Bargagli, R. Macro- and Microplastics in the Antarctic Environment: Ongoing Assessment and
Perspectives. Environments 2022, 9, 93. [CrossRef]
51. Siegfried, M.; Koelmans, A.A.; Besseling, E.; Kroeze, C. Export of Microplastics from Land to Sea. A Modelling Approach. Water
Res. 2017, 127, 249–257. [CrossRef]
52. van Franeker, J.A.; Law, K.L. Seabirds, Gyres and Global Trends in Plastic Pollution. Environ. Pollut. 2015, 203, 89–96. [CrossRef]
53. Wooddall, L.C.; Sanchez-Vidal, A.; Canals, M.; Paterson, G.L.; Coppock, R.; Sleight, V.; Calafat, A.; Rogers, A.D.; Narayanaswamy,
B.E.; Thompson, R.C. The Deep Sea is a Major Sink for Microplastic Debris. R. Soc. Open Sci. 2014, 1, 140317. [CrossRef]
54. Han, M.; Niu, X.; Tang, M.; Zhang, B.T.; Wang, G.; Yue, W.; Kong, X.; Zhu, J. Distribution of Microplastics in Surface Water of the
Lower Yellow River Near Estuary. Sci. Total Environ. 2020, 707, 135601. [CrossRef]
55. McEachern, K.; Alegria, H.; Kalagher, A.L.; Hanse, C.; Morrison, S.; Hastings, D. Microplastics in Tampa Bay, Florida: Abundance
and Variability in Estuarine Waters and Sediments. Mar. Pollut. Bull. 2019, 148, 97–106. [CrossRef]
56. Waite, H.; Donnelly, M.; Walters, L. Quantity and Types of Microplastics in the Organic Tissues of the Eastern Oyster
Crassostrea virginica and Atlantic Mud Crab Panopeus herbstii from a Florida Estuary. Mar. Pollut. Bull. 2018, 129, 179–185.
[CrossRef]
57. Gray, A.D.; Wertz, H.; Leads, R.R.; Weinstein, J.E. Microplastics in Two South Carolina Estuaries: Occurrence, Distribution, and
Composition. Mar. Pollut. Bull. 2018, 128, 223–233. [CrossRef]
58. Baechler, B.R.; Stienbarger, C.D.; Horn, D.A.; Joseph, J.; Taylor, A.R.; Granek, E.F.; Brandre, S.M. Microplastic Occurrence and
Effects in Commercially Harvested Finfish and Shellfish: Current Knowledge and Future Directions. Limnol. Oceanogr. Lett. 2020,
5, 113–136. [CrossRef]
59. Martinelli, J.C.; Phan, S.; Luscombe, C.K.; Padilla-Gamino, J.L. Low Incidence of Microplastic Contaminants in Pacific Oysters
(Crassostrea gigas Thunberg) from the Salish Sea, USA. Sci. Total Environ. 2020, 715, 136826. [CrossRef]
60. Wheat, E.; Ruesink, J.L. Commercially-Cultured Oysters (Crassostrea gigas) Exert Top-Down Control on Intertidal Pelagic Resources
in Willapa By, Washington, USA. J. Sea Res. 2013, 81, 33–39. [CrossRef]
61. Bougrier, S.; Geairon, P.; Deslous-Paoli, J.M.; Bacher, C.; Jonquieres, G. Allometric Relationships and Effects of Temperature on
Clearance and Oxygen-Consumption Rates of Crassostrea gigas (Thunberg). Aquaculture 1995, 134, 143–154. [CrossRef]
62. Coughlan, J. The Estimation of Filtering Rate from the Clearance of Suspensions. Mar. Biol. 1969, 2, 356–358. [CrossRef]
63. Dupuy, C.; Vaquer, A.; Lam-Hoai, T.; Rougier, C.; Mazouni, N.; Lautier, J.; Collos, Y.; Le Gall, S. Feeding Rate of the Oyster
Crassostrea gigas in a Natural Planktonic Community of the Mediterranean Thau Lagoon. Mar. Ecol. Prog. Ser. 2000, 205, 171–184.
[CrossRef]
64. Kobayashi, M.; Hofmann, E.; Powell, E.N.; Klinck, J.; Kusaka, K. A Population Dynamics Model for the Japanese Oyster,
Crassostrea gigas. Aquaculture 1997, 149, 285–321. [CrossRef]
65. Powell, E.; Hofmann, E.; Klinck, J.; Ray, S. Modeling Oyster Populations: I. A Commentary on Filtration Rate: Is Faster Always
Better? J. Shellfish Res. 1992, 11, 387–398.
66. Galimany, E.; Freeman, C.J.; Lunt, J.; Domingos, A.; Sacks, P.; Walters, L. Feeding Competition between the Native Oyster
Crassostrea virginica and the Invasive Mussel Mytella charruana. Mar. Ecol. Prog. Ser. 2017, 564, 57–66. [CrossRef]
67. Grizzle, R.E.; Greene, J.K.; Coen, L.D. Seston Removal by Natural and Constructed Intertidal Eastern Oyster (Crassostrea virginica)
Reefs: A Comparison with Previous Laboratory Studies, and the Value of In Situ Methods. Estuar. Coasts 2008, 31, 1208–1220.
[CrossRef]
68. Phuong, N.N.; Poirier, L.; Pham, Q.T.; Lagarde, F.; Zalouk-Vergnoux, A. Factors Influencing the Microplastic Contamination of
Bivalves from the French Atlantic Coast: Location, Season and/or Mode of Life? Mar. Pollut. Bull. 2018, 129, 664–674. [CrossRef]
69. Mak, C.W.; Tsang, Y.Y.; Leung, M.M.L.; Fang, J.K.H.; Chan, K.M. Microplastics from Effluents of Sewage Treatment Works and
Stormwater Discharging into the Victoria Harbor, Hong Kong. Mar. Pollut. Bull. 2020, 157, 111181. [CrossRef] [PubMed]
70. Bergman, M.J.; Donnangelo, L.J. Simulation of Freshwater Discharges from Ungauged Areas to the Sebastian River, Florida.
J. Amer. Water Res. Assoc. 2000, 36, 1121–1132. [CrossRef]
71. Kim, Y.T. Water Balance and Flushing Time in the Restricted Indian River Lagoon (IRL), Florida USA. Ocean Polar Res. 2003, 25,
75–87. [CrossRef]
Environments 2022, 9, 131 18 of 18

72. USGS Surface Water Data for the Nation. Available online: https://waterdata.usgs.gov/nwis/sw (accessed on 21 August 2022).
73. Law, K.L.; Morét-Ferguson, S.; Maximenko, N.A.; Proskurowski, G.; Peacock, E.E.; Hafner, J.; Reddy, C.M. Plastic Accumulation
in the North Atlantic Subtropical Gyre. Science 2010, 329, 1185–1188. [CrossRef]
74. Ji, Z.G.; Hu, G.; Shen, J.; Wan, Y. Three-Dimensional Modeling of Hydrodynamic Processes in the St. Lucie Estuary. Estuar. Coast
Shelf Sci. 2007, 73, 188200. [CrossRef]
75. Loosanoff, V.L. Some Aspects of Behavior of Oysters at Different Temperatures. Biol. Bull. 1958, 114, 57–70. [CrossRef]
76. Walters, L.; Phlips, E.; Badylak, S.; McClenachan, G.; Sacks, P.; Donnelly, M. Field Observations of the Negative Association
between Recruitment of the Eastern Oyster Crassostrea virginica and the Brown Tide Aureoumbra lagunensis in Mosquito Lagoon,
FL. Fla. Sci. 2021, 84, 81–91.
77. Walters, L.; Roddenberry, A.; Crandall, C.; Wayles, J.; Donnelly, M.; Barry, S.; Clark, M.; Escandell, O.; Hansen, J.C.;
Laakkonen, K.; et al. The Use of Non-Plastic Materials for Oyster Reef and Shoreline Restoration: Understanding What is Needed
and Where the Field is Headed. Sustainability 2022, 14, 8055. [CrossRef]
78. Morris, R.; Bilkovic, D.; Boswell, M.; Bushek, D.; Cebrian, J.; Goff, J.; Kibler, K.; La Peyre, M.; McClenachan, G.; Moody, J.; et al.
The Application of Oyster Reefs in Shoreline Protection: Are We Over-Engineering for an Ecosystem Engineer? J. Appl. Ecol. 2019,
56, 1703–1711. [CrossRef]
79. Morris, R.; Bilkovic, D.; Boswell, M.; Bushek, D.; Cebrian, J.; Goff, J.; Kibler, K.; Le Peyre, M.; McClenachan, G.; Moody, J.; et al.
Large-Scale Variation in Wave Attenuation of Oyster Reef Living Shorelines and the Influence of Inundation Duration. Ecol. Appl.
2021, 31, e02382. [CrossRef]
80. Beck, R.W. National Post-Consumer Plastics Recycling Report; American Plastics Council, Division of American Chemistry Council:
Arlington, VI, USA, 2004.
81. Delle Chiaie, K.R.; McMahon, F.R.; Williams, E.J.; Price, M.J.; Dove, A.P. Dual-Catalytic Depolymerization of Polyethylene
Terephthalate. Polym. Chem. 2020, 11, 1450–1453. [CrossRef]
82. Moore, C.J. Synthetic Polymers in the Marine Environment: A Rapidly Increasing, Long-Term Threat. Environ. Res. 2008, 108,
131–139. [CrossRef]
83. Jaffe, M.; Easts, A.J.; Feng, X. Polyester Fibers. In Thermal Analysis of Textiles and Fibers. The Textile Institute Book Series; Woodhead
Publishing: Sawston, UK, 2020; pp. 133–149.
84. Koh, L.D.; Cheng, Y.; Teng, C.P.; Khin, Y.W.; Loh, X.J.; Tee, S.Y.; Low, M.; Ye, E.; Yu, H.D.; Zhang, Y.W.; et al. Structures, Mechanical
Properties and Applications of Silk Fibroin Materials. Prog. Polym. Sci. 2015, 46, 86–110. [CrossRef]
85. Smith, N. The Intertidal Volume of Florida’s Indian River Lagoon. Fla. Sci. 1992, 55, 209–218.