Water Air Soil Pollut (2022) 233:455

https://doi.org/10.1007/s11270-022-05926-w

Microplastic in Sediments and Ingestion Rates in Three

Edible Bivalve Mollusc Species in a Southern Philippine
Estuary
P. S. P. Bonifacio · E. B. Metillo ·
E. F. Romano Jr

Received: 23 July 2022 / Accepted: 26 October 2022 / Published online: 4 November 2022
© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2022

Abstract Plastics are now a major environmental type of microplastic in terms of morphology, while
concern worldwide with their widespread contamina- black and blue are the dominant colors of microplas-
tion and accumulation. Microplastic particle (< 5 mm) tic particles. There were 1495 microplastic particles
is an emerging pollution issue as it is being detected (4.15 ± 3.37 particles/clam) present in the clam tis-
worldwide in aquatic and terrestrial ecosystems, but sues, of which polypropylene (PP) and rayon (RY)
relatively little is known in tropical regions. This polymers are the most common, whereas K. hiantina
study determined the (1) abundance of microplas- (707 particles) showed the highest amount of micro-
tics in sediment and (2) in situ and laboratory inges- plastics. The number of ATR-FTIR-confirmed poly-
tion rates of microplastics in three scarcely studied mer types in the wild clams is greater than that in the
tropical bivalve mollusc species (Donax sp., Meretrix sediments. The study reveals abundant microplastics
meretrix, and Katelysia hiantina) in Panguil Bay, in sediments and in the three species of bivalve indi-
Southern Philippines. A total of 2258 microplastic viduals from the wild. All clams ingested low-density
particles (62.72 ± 18.31 items/m2) were found on the polyethylene (LDPE) microplastic particles in the
sediment samples. Filament/fiber is the most abundant laboratory. The mean number of LDPE microplas-
tic particles ingested by clams is 4.62 ± 2.40 parti-
cles/clam/7days, with the highest value observed in
Supplementary Information The online version K. hiantina. Additionally, Donax sp., M. meretrix,
contains supplementary material available at https://​doi.​
org/​10.​1007/​s11270-​022-​05926-w.
and K. hiantina could ingest high densities of 40–60-
μm microplastic particles.
P. S. P. Bonifacio
Department of Biological Sciences, College of Science Keywords Bivalve mollusc · ATR-FTIR analysis ·
and Mathematics, Western Mindanao State University,
Microplastics · Panguil Bay · Sediments · Ingestion
7000 Zamboanga City, Philippines
rate
E. B. Metillo (*)
Department of Marine Science, College of Science
and Mathematics, Mindanao State University - Iligan
1 Introduction
Institute of Technology, 9200 Iligan City, Philippines
e-mail: [email protected]
Plastics are widely used in different applications
E. F. Romano Jr because they are lightweight, durable, and economi-
Chemistry Department, College of Arts and Sciences,
cal. According to PlasticsEurope (2020 as cited in
Negros Oriental State University, 6200 Dumaguete City,
Philippines Mai et al., 2022), global plastic production reached

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368 million tons in 2019. Moreover, due to the market 2015), and their small dimension makes them availa-
demand for plastic products, it is projected that plastic ble for ingestion by a wide range of marine organisms.
production may reach 600 million tonnes by 2025 and Besides entanglement and ingestion of macro debris
may exceed 1 billion tonnes by 2050 (FAO, 2017). by large vertebrates, microplastics are accumulated
Around 6300 metric tonnes (MT) of plastic waste is by planktonic and invertebrate organisms, being trans-
generated annually, out of which 9% is recycled, 12% ferred along food chains through the consumption of
is incinerated, and the remaining 79% gets accumu- lower trophic level prey (Avio et al., 2016; Farrell and
lated in natural environments and ultimately ends up Nelson, 2013). Primary sources of microparticle-sized
in seas and oceans (Geyer et al., 2017). The COVID- plastics refer to the pellet form of plastics used as
19 pandemic may have increased microplastic pollu- feedstock in the plastic industry and/or personal care
tion due to the global production of personal protec- products. In contrast, microplastics derived from sec-
tive equipment (PPE), gloves, face masks, and face ondary sources are a result of fragmentation and deg-
shields in billion pieces (Lee and Kim, 2022). The radation of larger plastic materials (Cole et al., 2011;
common sources of plastic pollution in the terrestrial Arp et al., 2021). The majority of microplastics in the
and marine environments come from the packaging oceans are secondary products derived from the deg-
materials that we use every day, which are made up of radation and fragmentation of mesoplastics or larger
polyethylene (PE), polypropylene (PP), polystyrene fragments (Gregory and Andrady, 2003).
(PS), polyethylene terephthalate (PET), polyvinyl In general, the bioavailability of microplastics
chloride (PVC), and polyamide (PA) (Andrady, 2011; and nanoplastics 0.001 to 0.1 μm (FAO, 2017) may
Avio et al., 2016). depend on their size, density, shape, and surface
Plastic wastes are potential threat to marine organ- charges which will affect their behavior in seawater,
isms. For example, ingestion of plastic debris or leading to agglomeration, resuspension, and settling.
entanglement has been recorded in 44–50% of all Moreover, their uptake, disposal, and bioaccumula-
seabirds, sea snakes, sea turtles, penguins, seals, sea tion by marine organisms are influenced by their feed-
lions, manatees, sea otters, fish, crustaceans, and half ing behavior, with benthic detritivores and suspension
of all marine mammals (Kühn et al., 2015). Moreo- feeders representing more susceptible target species
ver, these plastic wastes contribute to the formation of (Wright et al., 2013). Laboratory studies conducted
a new contaminant called microplastics. Reports indi- show that microplastics can be ingested by a range
cate a global increase and widespread distribution of of marine organisms when particles are mistaken
microplastics in the environment, thereby increasing for food (Browne et al., 2008; Boerger et al., 2010;
microplastic’s bioavailability to various organisms. Lusher et al., 2013).
This scenario suggests that microplastics potentially In addition to direct physical impacts from the
threaten wildlife, natural ecosystems, and human microplastic itself, ingested plastic debris may act
health (Endo et al., 2005; Teuten et al., 2009; Rocha- as a medium to concentrate and transfer chemi-
Santos and Duarte, 2015; Arp et al., 2021). cals and persistent, bioaccumulative, and toxic sub-
Microplastics, typically defined as plastic pieces stances (PBTs), such as polychlorinated biphenyls
that are < 5 mm in size (Barnes et al., 2009), although (PCBs) to organisms (Eerkes-Medrano et al., 2015).
a recent definition suggests to consider fragments Different particles polymers, like PVC, PE, PP, and
smaller than 1 mm (Browne et al., 2015), have been PS, were shown to have a high sorption capacity for
documented in aquatic systems on all seven conti- dichlorodiphenyltrichloroethane (DDT), polycyclic
nents, in both freshwater and marine environments aromatic hydrocarbons (PAHs), hexachlorocyclohex-
(Barnes et al., 2010), along beaches (Browne et al., anes (HCH), and chlorinated benzenes (Bakir et al.,
2011, Yabanlı et al., 2019), in sediments (Claessens 2012; Lee et al., 2014). This diversity of synthetic
et al., 2011; Reed et al., 2018), in biota (bivalve mol- substances places human beings at high risk for can-
luscs, zooplankton, and fish) (Rashid et al., 2021; cer and debilitating diseases (Dewailly et al., 1989;
Ding et al., 2021; Wootton et al., 2021), and in the Lohman and Muir, 2010).
water column itself (Eriksen et al., 2013). Microplas- The estuarine water of Panguil Bay, an important
tics are barely visible microlitter in the form of small fishing ground in Southern Philippines, supports
fragments, fibers, and granules (Desforges et al., three species of exploited bivalve molluscs, namely,

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the wedge clam, Donax sp., Asian hard clam, Mer- NE monsoon months (from October to March), but
etrix meretrix, and the surf clam, Katelysia hiantina lower (3.3–29.7) in the warmer (30–32 °C) south-west
(Jimenez et al., 2009). Microplastic contamination in (SW) monsoon months of June through to September.
habitats and ingestion in bivalve mollusc food spe- Higher mean dissolved oxygen values (7.1–7.9 mg
cies are poorly studied in the tropical region (Espiritu ­L–1) occur in November and August, compared with
et al., 2019). Thus, this study aimed to estimate the those (6.5–6.6 mg L ­ –1) in February and May. The pH
distribution and abundance of microplastic materi- (8.8–9.2) is typical of normal seawater in February,
als in the sediment habitat of the three bivalve spe- but slightly acidic, typical of a mangrove estuary, in
cies, quantify the abundance of microplastics in other months. Dissolved nitrate was generally lower
three bivalve mollusc species (= in situ ingestion), (0.4–0.7 mg ­L–1) in rainy SW monsoon months com-
and determine the ingestion rates of the three spe- pared with the warmer months of February through to
cies on low-density polyethylene microplastic in the May (0.4–0.9 mg ­L–1). The lowest concentrations of
laboratory. dissolved orthophosphate were recorded in November
(0.1–0.2 mg ­L–1), but higher in rainy months during
SW monsoon (0.1–0.4mg ­ L–1). Heavy rainfall and
2 Materials and Methods strong winds during NW monsoon coincided with
highest concentrations (0.8–2.9 x ­ 102 mg ­ L–1) of
2.1 Sampling Area total suspended solids throughout the bay.” The most
abundant commercially exploited species in the bay
Sediment and bivalve mollusc samples were collected are invertebrates with molluscs contributing to 10%
from four different sites (Baroy [site 1], Tangub [site (Jumawan et al., 2021).
2], Kapatagan [site 3], Cabgan [site 4]) in the estu- Plastic materials like fishing net, rope, plastic bag,
arine Panguil Bay (Fig. 1). Panguil Bay is about 41 adhesive tape, styrofoam, tire cord, wire, and cable
km long and is flanked by 10 municipalities and two insulation, and food and drink wrappers were found
cities belonging to three provinces (Lanao del Norte, in the sampling area which could be some of the
Zamboanga del Sur, and Misamis Occidental) and possible sources of plastic pollutants in Panguil Bay
three administrative regions (Regions 9, 10, and 12). (Bonifacio, 2021).
There are 11 towns and 78 coastal barangays sur-
rounding Panguil Bay (Jumawan et al., 2020). 2.2 Collection and Quantification of Microplastics in
Panguil Bay description, hydrography, and water Sediments
circulation were reported by Canini et al. (2013) and
Metillo et al. (2015). As reported, “Panguil Bay is a Sampling of sediments was conducted at three tran-
south-western inlet of the greater Iligan Bay, and with sect lines with three quadrats located 10 m away from
an average of 5 m, the depth gradually decreases from each other for each area, specifically in the intertidal
the mouth to the inner portion, but a narrow and deep zone. The sampling quadrat was 1 × 1 m. A metallic
lengthwise channel in the middle occupies three- shovel was used to collect sediments at 5 cm depth.
quarters of the total length of the bay. The volume of The sediments were sealed in a plastic container
exchanged water daily is about 22 % of the total vol- which was rinsed in the laboratory with seawater
ume of 1.1 ­km3. The remaining mangrove forest has before use. Sediments were transported back to the
an area of 27.3 ­km2, mostly concentrated in the inner laboratory, dried, and kept at room temperature until
parts of the bay. Rainfall on the catchment area brings extraction (Barasarathi et al., 2014; Besley et al.,
runoff into Panguil Bay via 29 major rivers and 46 2016). Dried sediment samples were sieved through
minor tributaries. Variation of hydrographic param- 0.45 mm mesh to separate macroplastics. Microplas-
eters is primarily influenced by monsoon. Strong tics in 2 kg sediment samples were extracted using
north-east (NE) monsoon winds prevail over Panguil the method adapted from Besley et al. (2016) with
Bay from November to March but are strongest from minor modifications and additional changes. A fully
January to February. Weak, variable winds typify the saturated salt solution was prepared by dissolving
intermonsoon months of April and May. Salinity is 358.9 g of sodium chloride (NaCl) in 1 L of deminer-
generally higher (10–31.7) in the cooler (24–32 °C) alized water. The salt solution was filtered to remove

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1
4
Philippines

Fig. 1  Sampling sites for microplastics in sediments and bivalve molluscs (Donax sp., Meretrix meretrix, and Katelysia hiantina) in Baroy (1), Tangub (2), Kapatagan (3), and
Cabgan (4) in Panguil Bay, Southern Philippines
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impurities. Microplastic was extracted by density Individuals of three species were randomly placed in
separation, where dry sand was combined with a satu- individual and identical 500-mL glass beakers with
rated salt solution. One hundred (100) g of dried sand 400 mL of the microplastic-seawater mixture fol-
was added to 400 mL of fully saturated salt solution lowing a setup of three bivalve mollusc species × 10
and was stirred for 5–10 min. The sediments were replicate individuals of each species. Microplastic-
then allowed to settle for not less than 3 days or until seawater mixture was prepared by mixing 1.5 g of
the water became clear. The supernatant was poured 40–60-μm LDPE microplastics and 1 L of filtered
into a vacuum filtration system and filtered through seawater. No sediments were added, and portable
Whatman filter paper grade 1 (11 μm pore size). Pro- oxygen pumps were used to provide aeration. The use
cedures were repeated two times for each sample. of plastic material was avoided while experimenting.
Collected particles in the filter paper were examined The exposure ran for 7 days. The water was changed
using a stereo microscope, compound microscope, in each glass beaker every 24 h with LDPE micro-
and ATR-FTIR spectroscopy. Blanks consisting of plastics. Clams were not fed to avoid any interaction
salt solution only were handled the same way as sam- of microplastics and food, but animals remained alive
ples to assess laboratory contamination. through the microplastic exposure period. After 7
days, ingestion rates of clams were measured using
2.3 Source of Polyethylene Microplastic the same acid digestion protocol for the wild-based
clams. Usually, whole organisms (Lusher et al.,
Low-density polyethylene (LDPE) particles were 2017b) are digested or analyzed in laboratory studies.
obtained from Beijing Yan Lin Fu Chemical Sales
Center (China) and were processed and size-sorted 2.6 Acid Digestion of Clams from the Wild and
into 40–60-μm microplastics through grinding and Laboratory Experiment
sieving by milling machine, blender, and metallic
sieves. LDPE microplastic sizes were verified under a Microplastics from wild clams and LDPE synthetic
compound microscope. microplastics from clams in the laboratory experi-
ment were determined using the digestion protocol
2.4 Collection and Laboratory Maintenance of adapted from Claessens et al. (2013) with minor
Samples modifications. Clams were refrigerated (1 °C)
before and during preliminary processing and were
A total of 840 bivalve mollusc individuals were col- frozen until acid digestion could take place. Dur-
lected in Panguil Bay. The three bivalve mollusc spe- ing freezing, clams gape, creating a potential route
cies (the wedge clam, Donax sp.; Asian hard clam, for contamination. Clams were bound with elas-
Meretrix meretrix; and the surf clam, Katelysia hian- tic bands while in the freezer to maintain pressure
tina) were collected in the same four sampling sites on both sides of the shell to avoid contamination
of the Panguil Bay. After collection, 360 mollusc (Davidson and Dudas, 2016). Clams were not depu-
individuals for acid digestion were refrigerated, and rated in order to include microplastics that had been
480 mollusc individuals intended for polyethylene both recently ingested and possibly translocated to
ingestion rate study were transferred to the laboratory, tissues (Mathalon and Hill, 2014; Li et al., 2015).
where they were acclimatized in filtered seawater for Clamshells were thoroughly rinsed with water and
7 days at constant aeration, with a photoperiod of 12 then removed using a scalpel and forceps. The soft
h light and 12 h darkness. tissues were weighed separately. Clam tissues were
digested in 10–15 mL of 69% nitric acid overnight
2.5 Laboratory Feeding at room temperature in a beaker/flask, followed by
⁓2 h of boiling (⁓100 °C). Digestions were com-
Live individuals were scrubbed to remove any plete when there was no visible organic material
organisms from their shells. The laboratory expo- remaining, and the solution was clear and yel-
sure method was adapted from Avio et al. (2015) low. The resulting mixture was diluted (1:10 v/v)
and Ribeiro et al. (2017) with minor modifications. with warm (⁓80 °C) deionized water and vacuum

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filtered over 3-µm pore-sized filter paper immedi- 2.8 Contamination Control

ately after dilution. All beakers and Buchner fun-
nels were rinsed with deionized water before diges- Contamination control measures were strictly imple-
tion and throughout to reduce the amount of sample mented and followed all throughout the experimenta-
lost during transfer stages. Filter papers were then tion. The use of plastic materials was minimized and
examined under a stereomicroscope (10–40× mag- all equipment were rinsed with distilled water before
nification) for microplastic particles. Individual every test. Container walls were washed frequently
microplastic samples were further examined under with distilled water during the extraction of micro-
a compound microscope (10–100× magnification) plastics to avoid losing samples. Laboratory room
when necessary. was cleaned before and after daily experimentation.
Cotton coats were worn and the number of people
was minimized inside the laboratory. Control samples
2.7 Microplastic Identification were constantly checked for contamination.

The simplest method in the identification of micro-

2.9 Statistical Analysis
plastics is visual identification, a time-consuming but
the most appropriate technique when dealing with a
Plastic particle concentrations were represented as
high volume of samples and limited resources (Craw-
mean values ± standard deviation (SD). Kruskal-
ford and Quinn, 2017). Microplastics were identified
Wallis H Test was used to evaluate microplastic
by color, structure, and type after clearing from debris,
abundance in sediments and clams among sampling
salt crystals, and sand grains. Most microplastics
sites, and LDPE exposed clams among sampling
were flexible, will not break, and often spring back
sites and mollusc species since data did not meet the
when prodded. A fine tweezer was used when prod-
assumption of normality (Shapiro-Wilk W Test) and/
ding without damaging the microplastics. Microplas-
or homogeneity of variance (Levene’s Test). Statisti-
tics could be covered by debris, salt crystals, and sand
cal significance was defined at p < 0.05. All statistical
grains, making them difficult to see and identify. The
data analyses were performed using the IBM SPSS
chemical composition of microplastic samples from
Statistics version 23.
sediments and bivalves was determined using Fourier-
transform infrared spectroscopy (Perkin Elmer Spec-
trum Two, USA) attenuated total reflectance mode (8
to 14 scans, 4 ­cm−1 resolution, 4000 ­cm−1 to 450 ­cm−1 3 Results and Discussion
wave number range), a standard analytical technique
a. Abundance and distribution of microplastics in
for identifying the chemical composition of samples
sediments and wild clams
larger than 0.5 mm (Biginagwa et al., 2015). The fol-
lowing criteria were used to define a plastic particle
Microplastics found in the sediments and wild
through visual examination (Noren, 2007):
clams were categorized into fragments, filament/fiber,
and foam (Fig. 2). A total of 2258 microplastic par-
a. No cellular or organic structures are visible in the
ticles in the sediments were visually identified under
plastic particle/fiber.
the microscope. Filament/fiber (56%) was found to
b. If the particle is a fiber, it should be equally thick,
be the predominant type of microplastic, followed by
not taper towards the ends, and have a three-
fragments (42%) and foam (2%) being the lowest. On
dimensional bending (not entirely straight fibers,
the other hand, a total of 1495 microplastic particles
which indicates a biological origin).
were found on clams in which filament/fiber (65%)
c. Clear and homogeneously colored particles (blue,
was the predominant type of microplastic particles,
red, black, and yellow).
followed by fragments (35%). No styrofoam was
d. If it is not obvious that the particle/fiber is
found in all the clam samples. Filaments/fibers are the
colored, i.e., if it is transparent or whitish, it shall
most abundant microplastics since apart from plastic
be examined with extra care in a microscope
bags and bottles, packaging straps and fishing lines in
under high magnification.

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Fig. 2  Microplastic cat-

egory in sediments and wild
clams

oceans are the most common plastic pollution items particular concern since they could easily be mistaken
(Blettler et al., 2018). Additionally, polyolefins (PE for food by marine organisms and seabirds (Thomp-
and PP) and nylons are primarily used in fishing gear son et al., 2004; Browne et al., 2008; Gregory, 2009).
applications and are commonly detected in coastal The highest number of microplastics in sediments
areas (Klust, 1982; Timmers et al., 2005). was found in site 4 (75.89 ± 22.88 items/m2), fol-
Different colored microplastic filaments/fibers lowed by site 1 (63.78 ± 10.65 items/m2), and site 2
were found in the sediment and wild clam samples (61.67 ± 17.19 items/m2) while site 3 (49.56 ± 11.93
(Fig. S1). Most of the filaments/fibers found were items/m2) had the lowest number (Fig. 4A). The
≤ 1 mm in size, measuring them along their longest abundance of microplastic particles observed in sedi-
dimension. Colors of microplastic particles in sedi- ments was similar within and among sampling sites.
ments and wild clams were black, blue, brown, green, Hence, we can assume that the abundance of micro-
orange, red, transparent, violet, white, and yellow plastic particles in sediments in the sampled areas
(Fig. 3). In sediments, black (28%) and blue (31%) and perhaps the entire Panguil Bay is homogenous.
particles are the predominant colors. The same domi- While prevailing currents and waves may cause dif-
nant colors were found in clams—black (37%) and ferences in the distribution of macroplastics, pro-
blue (14%). In this study, black and blue accounted cesses involved in microplastic distribution are less
for more than half of the colors of microplastics in well-known. Still, they can be influenced by particle
both sediment and clams, but color and shape, when distribution in the water column, settling rate, parti-
describing and quantifying microplastics, are still cle redistribution via hydrodynamics, and particle
in question as they introduce subjectivity due to the aggregation or animal activities (Browne et al., 2010).
process of visual sorting with microscopy (Baruah The mean abundance of sediment microplastics in
et al., 2022). Colored microplastics are the most com- the sampling sites is 62.72 ± 18.31 items/m2. This is
mon in water, accounting for 50.4–86.9% of the total higher compared to that reported in a Brazil estuary
microplastics (Wang et al., 2017). Studies in China with 60 items/m2 (Ivar do Sul et al., 2009) but lower
indicated that transparent and blue microplastics are compared to those in Portugal with 133.3 items/m2
the most abundant microplastics detected in biota (Martins and Sobral, 2011); South Korea with 913
(Fu et al., 2020). Recently, grouping microplastics items/m2 (Heo et al., 2013); St. Lawrence River, Can-
into four obvious colors (transparent, black, white, ada with 13,832 ± 13,677 items/m2 (Castañeda et al.,
and colored) instead of evaluating other more con- 2014); and Pearl River, Hongkong with 5595 items/
troversial colors (e.g., yellow, green, blue) has been m2 (Fok and Cheung, 2015).
suggested by Jiang et al. (2019). However, the vari- Microplastic particles present in wild clams (N
ations in size, shape, and color in microplastics are a = 30 of each of the species of clams: Donax sp. ≤

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Fig. 3  Abundance of
microplastic particles in
sediments (A) and wild
clams (B) from Panguil
Bay, Southern Philippines.
The number of microplas-
tic particles from each
sampling site (N = 30
per species) was visu-
ally determined. (C) The
number of microplastics per
clam ranged from 0 to 21
particles (Table S1)

1 g, Meretrix meretrix 3–5 g, and Katelysia hinatina particles per clam (Fig. 4C) reveals that the high-
3–7 g per site) are presented in Fig. 4B. The highest est mean value was recorded in K. hiantina (5.89 ±
total number of microplastic particles per individual 4.23), followed by M. meretrix (4.16 ± 2.86) and,
clam was observed in K. hiantina (707), followed by lastly, Donax sp. (2.41 ± 1.47). Considering all the
M. meretrix (499), and the lowest in Donax sp. (289). clam species (N = 360), the grand mean of particles
Meanwhile, sites 2 and 3 showed the highest number per clam is 4.15 ± 3.37, with filament/fiber being the
of microplastic particles ingested by one individual of most abundant shape. The abundance of microplas-
M. meretrix (19) and K. hiantina (21), respectively. tic particles observed in clams was statistically sig-
The lowest number of microplastics was found in nificant within and among sampling sites and mollusc
Donax sp. (8) from site 4. All bivalve mollusc species species. Hence, we can assume that the abundance
have samples that have zero microplastics found after of microplastic particles among bivalve mollusc spe-
digestion. Comparison of mean values of microplastic cies varies, which could be attributed to contrasting

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Fig. 4  Types of micro-

plastic polymer found in
sediments (A) and wild
clams (B) of Panguil Bay,
Southern Philippines.
Polymer types: acryloni-
trile butadiene styrene
(ABS), cellulose acetate
(CA), polyethylene (PE),
polyethylene terephthalate
(PET), polypropylene (PP),
polystyrene (PS), polyvi-
nyl chloride (PVC), azlon
(AZ), ethylene vinyl acetate
(EVA), nylon-6 (PA6), phe-
noxy resin (PR), polysty-
rene (PS), and rayon (RY).
Unclassified particles have
a correlation of < 0.7

feeding mechanisms and rates (ability to reject inedi- wild clam species (Fig. 5B). The number of FTIR-
ble particles) among species (Van Cauwenberghe and confirmed polymers is greater in the wild clams than
Janssen, 2014). in sediments. These polymers are azlon (AZ), ethyl-
Out of the 2258 visually identified particles from ene vinyl acetate (EVA), nylon-6 (PA6), polyethylene
the four sediment sampling sites, 56 particles (14 par- (PE), polyethylene terephthalate (PET), polypropylene
ticles × 4 sites) were randomly selected and further (PP), phenoxy resin (PR), polystyrene (PS), polyvinyl
validated via FTIR spectroscopy (Fig. 5A). The iden- chloride (PVC), and rayon (RY). The top three poly-
tified polymer types include acrylonitrile butadiene mers found in clams are RY (62), PET (37), and PP
styrene (ABS), cellulose acetate (CA), polyethylene (21), while PS (1), and PVC (1) are present in the
(PE), polyethylene terephthalate (PET), polypropylene smallest amount. Of the 175 samples, 22 were unclas-
(PP), polystyrene (PS), and polyvinyl chloride (PVC) sified polymers due to poor correlation values.
(Fig. S2). The top three polymers are PP (23), PVC Chemical composition is the most basic criterion
(12), and PE (10). A total of 175 particles were suc- for defining microplastic pollution (Zhang et al.,
cessfully identified as plastic polymers from the three 2020). Plastics are synthetic polymers made from

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Fig. 5  Proportion of
microplastics in terms of
color in sediments (A) and
wild clams (B) found in the
habitat of the three bivalve
mollusc species in Panguil
Bay, Southern Philippines

various compounds with different characteristics, microplastic contamination since the widely used
and FTIR and Raman spectroscopy are the highly microscopy, photography, and spectroscopy may still
recommended equipment to identify them (Hidalgo- result in unrealistic contamination estimates (Kroon
Ruz et al., 2012). Larger microplastic particles (> et al., 2018). In fact, not all microplastics were sent
500 μm) can be analyzed by ATR-FTIR (Tagg et al., for FTIR analyses since some were too small in size
2015; Ivleva et al., 2017). For smaller particles, FTIR for FTIR-ATR analysis. If not careful, the number
must be combined with an optical microscope, the identified and the number analyzed may have discrep-
so-called micro-FTIR technique (Imhof et al. 2012; ancy due to the particles being lost during the inspec-
Zhang L. et al. 2020), which can analyze 10 μm sizes tion, microscopic photography, and transferring of the
via reflection and transmission mode (Simon et al., microplastics to containers for spectroscopy. There is
2018). In this study, background scans were done always an element of subjectivity and potential bias
before analysis, and each spectrum was evaluated during visual identification. The effectiveness of vis-
manually based on the peaks’ position. Samples with ual identification is dependent on the size of the item
low correlation (< 0.6) were excluded from the inter- being scrutinized and becomes more considerably dif-
pretation of the final results (Bucol et al., 2020). In ficult as the size of the item decreases. Microscopy
the present study, a total of 12 plastic polymers were techniques though, which were done in this study, can
identified via ATR-FTIR spectroscopy: ABS, AZ, confirm the absence or presence of cellular structures
CA, EVA, PA6, PR, PE, PET, PP, PS, PVC, and RY, and improve visual identification skills (Crawford and
half of which is included in the most common types Quinn, 2017).
of polymers found in the marine environment. These
plastic polymers are commonly used in food packag- b. Laboratory feeding experiment
ing and fishing activities. The most common plastic
polymers in the marine environment are PE, PP, PS, All individuals (100%) of Donax sp., Meretrix
PVC, PA, PVA, and PET (Hidalgo-Ruz et al., 2012; meretrix, and Katelysia hiantina had ingested low-
Avio et al., 2016; Lusher et al., 2017a). density polyethylene (LDPE) microplastics after 7
Comparisons of microplastics have been difficult days of exposure period (Table 1). The highest num-
and limited to a few studies due to the difference in ber of LDPE microplastics ingested by Donax sp.,
units used in other reports, a drawback observed in M. meretrix, and K. hiantina after 7 days were 6,
current microplastic research (Wright et al., 2013; 14, and 13, respectively. The highest total number
Van Cauwenberghe et al., 2015), as well as plas- of LDPE microplastics was ingested by K. hiantina
tics in general. Some units cannot be transformed (740) followed by M. meretrix (608), and the lowest
between each other as they are measured in differ- in Donax sp. (316). The mean LDPE microplastic
ent dimensions (Yu et al., 2020). Additionally, there particle ingested by the three clam species was 4.62
are no universally accepted methods for quantifying ± 2.40 particles/clam/7days. A total of 1664 LDPE

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Water Air Soil Pollut (2022) 233:455 Page 11 of 16 455

Table 1  Amount of Mollusc Sampling site N Min Max Number of Sum Mean ± SD
low-density polyethylene microplas- (particles/clam/7days)
(LDPE) microplastic tic
particles ingested by
the four clam species in Donax sp. Baroy 30 2 5 89 316 2.63 ± 0.99
laboratory experiment for a
Cabgan 30 1 3 63
duration of 7 days
Kapatagan 30 1 4 77
Tangub 30 1 6 87
Meretrix meretrix Baroy 30 1 9 134 608 5.67 ± 2.37
Cabgan 30 2 7 139
Kapatagan 30 2 13 169
Tangub 30 3 14 166
Katelysia hiantina Baroy 30 4 9 170 740 6.17 ± 2.05
* Cabgan 30 3 10 144
The minimum and
maximum values are Kapatagan 30 4 8 182
the numbers of particles Tangub 30 6 13 244
obtained in each clam Total 360 1664 4.62 ± 2.40
sample

microplastic particles were ingested by 360 bivalve the relationship between plastic uptake and egestion
mollusc individuals. Meanwhile, the mean number up to date (Van Cauwenberghe and Janssen, 2014).
of LDPE microplastic particles ingested by the three In the study of Graham et al. (2019), it was reported
clam species was 4.62 ± 2.40 beads/clam/7days. The that Pacific oysters showed an efficient egestion rate,
study revealed that Donax sp., Meretrix meretrix, and egesting 84.6 ± 2% of the microplastic particles taken
Katelysia hiantina could ingest 40–60-μm microplas- up, which is slightly higher compared to the 74.5%
tic particles. In the study of Van Cauwenberghe et al. egestion rate post-depuration in a study conducted
(2015), the mussel Mytilus edulis can retain in their by Van Cauwenberghe and Janssen (2014), while
tissues particles ranging from 10 to 30 μm. It means only the 15.4 ± 2% of beads taken up were retained
that organisms will most likely ingest when micro- within the shell cavity. Ingestion and egestion rates of
plastic particles are readily available in their present microplastics vary among bivalve mollusc species as
environment, especially in high amounts. Results both are influenced by particle selectivity (Ward et al.
showed a significant difference within and among 2019), food or particle abundance, and other environ-
sampling sites and mollusc species exposed to LDPE mental conditions (Chae and An, 2020).
microplastic particles for 7 days. Such difference rein-
forces our main finding on the differences in micro-
plastic accumulation among the three clam species, 4 Conclusion
invoking a similar explanation of contrasting feeding
behavior among bivalve filter-feeding species. The abundance, distribution, and characteristics of
Other bivalve mollusc studies focus on the micro- microplastic particles in the marine environment and
plastic accumulation, translocation, and effects on the the aquatic organisms’ ingestion capability are essen-
organisms’ tissues (liver, gut, gills, and brain) by car- tial for evaluating microplastics’ potential effects and
rying out different biological analyses (e.g., Browne impacts on ecosystems. Sediment and clam sam-
et al., 2008; von Moos et al., 2012; Avio et al., 2015; ples obtained from the different sites of Panguil Bay,
Sussarellu et al., 2015; Van Cauwenberghe et al., Southern Philippines, were positive for microplastics,
2015). Additionally, ingestion and egestion in marine predominantly black and blue colored filament/fiber
organisms are important processes that can poten- and fragment particle type. Using ATR-FTIR spectros-
tially influence the cycling and transformation of copy, 7 polymer types were identified in sediments—
secondary pollution by microplastics (Browne et al., ABS, CA, PE, PET, PP, PS, and PVC, whereas 10
2011). However, there is still limited knowledge on polymers were identified in clams—AZ, EVA, PA6,

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 455 Page 12 of 16 Water Air Soil Pollut (2022) 233:455

PE, PET, PP, PR, PS, PVC, and RY. These polymer Declarations
materials came from fishing nets, rope materials, and
food and drink wrappers/packages. All clams exposed Ethics Approval Not applicable
to low-density polyethylene (LDPE) microplastic par-
ticles were found to have ingested the microplastic. Consent to Participate Not applicable
The highest number of LDPE microplastic particles
ingested within 7 days of laboratory exposure is 14, Consent for Publication Not applicable
while the lowest is 1. Further studies in microplastics,
Conflict of Interest The authors declare no competing interests.
especially in the Philippines, are needed to establish
standardized sampling, extraction, and identification
techniques. Also, food-related effects from microplas-
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