J. Eng. Technol. Sci., Vol. 54, No.

5, 2022, 220506

Microplastics in Wild Clams Harvested from Coastal

Waters of Lamongan, Indonesia
Muhammad Arif Asadi1,2*, Feni Iranawati1, Fairus Nafidya1, Supriyadi Supriyadi3
& Avijit Talukder4

1
Marine Science Department, Brawijaya University, Faculty of Fisheries and Marine
Science, Jalan Veteran Malang No. 16, Malang, East Java, 65145, Indonesia
2
Coastal Resilience and Climate Change Adaptation Research Group, Brawijaya
University, Jalan. Veteran Malang No. 16, Malang, East Java, 65145, Indonesia
3
Department of Capture Fisheries, Republic of Indonesia Defence University, Indonesia
Peace and Security Center, Sentul, 16810, Indonesia
4
Department of Marine Bioresource Science, Faculty of Fisheries, Chattogram
Veterinary and Animal Sciences University, Chattogram, 4225, Bangladesh
*E-mail: [email protected]

Highlights:
 A first assessment of microplastics present in different species of wild clams in
Indonesia.
 Almost all clams (99.23%) had ingested microplastics.
 The average concentration of microplastics in all clam samples was 3.5 ± 2.8
items/individual and 3.2 ± 2.52 items/g.
 Fibers dominated the microplastics present in the wild clams (80.77%).

Abstract. Contamination by microplastics has been reported in a wide array of

marine organisms, including bivalve mollusks. This study aimed to investigate the
occurrence and abundance of microplastics in bivalves from coastal waters of
Paciran, Lamongan, East Java, Indonesia. The high human population and
anthropogenic activities in the area may lead to contamination by microplastics in
marine organisms. Microplastics were recovered from the soft tissues of three
species of wild clams, Gafrarium tumidum, Anadara antiquata, and Venerupis
philippinarum. For each species, the microplastics found were grouped based on
type and size. The extraction of microplastics was done using 30% H2O2, while
the flotation method with NaCl was used to separate the dissolved liquid from the
clam’s soft tissues. Microplastic ingestion was found in 99.23% of the clam
samples with an average concentration of 3.5 ± 2.8 items/individual and 3.2 ± 2.52
items/g. V. philippinarum had the highest microplastics concentration (4.9 ± 2.80
items/g and 5.6 ± 3.22 items/individual). Microplastic types from all samples were
dominated by fibers (80.77%), where 57% of them were less than 200 µm in size.
The ubiquitous contamination of microplastics in clams from the coastal waters of
Lamongan, Indonesia is a serious concern for marine food webs and human health.

Received August 31st, 2021, 1st Revision October 27th, 2021, 2nd Revision April 22nd, 2022 Accepted for
publication May, 18th, 2022.
Copyright ©2022 Published by ITB Institute for Research and Community Services, ISSN: 2337-5779,
DOI: 10.5614/j.eng.technol.sci.2022.54.5.6
 Microplastics in wild clams of Indonesia

Keywords: 30% H2O2; fibers; fragments; Gafrarium tumidum; microplastics; Venerupis

philippinarum.

1 Introduction
The widespread use of plastic products drives the mass production of plastics [1-
3]. Since the 1960s, global plastics production has been growing steadily by
approximately 8.7% annually [4]. It was estimated that over eight million metric
tons of plastics enter the oceans annually [5], and plastics are now found in all
marine environments [6], from coastal areas [3,7] to deep sea sediments and polar
seas [8]. Once present in the marine environment, natural forces like wave action
and ultraviolet radiation will eventually degrade plastic waste into microplastics
(defined as plastic particles less than 5 mm in diameter) [5].

The first paper to describe microplastics was Buchanan [9], which reported
synthetic fibers with densities of up to 105/m3 in water samples from the North
Sea. Carpenter & Smith [10] noted a high density of polystyrene pellets in coastal
waters off southern New England. In the early 2000s, confirmation that
microplastics in the form of fragments and fibers are a ubiquitous form of marine
pollution has stimulated public awareness and research interest in microplastics
[11].

Microplastics as significant marine pollutants have been found in multiple marine

organisms [1,12,13]. The adherence and ingestion of microplastics have been
reported in a wide range of planktonic organisms, such as krill, zooplankton,
copepods, and other planktonic crustaceans [14-16]. Microplastics have also been
identified in fish, turtles, marine mammals, large crustaceans, and echinoderms
[17].

Mussels and other bivalves have been reported to contain and ingest multiple
types of microplastics in their tissues [12,18-20]. Fibers were the most common
types of microplastics in bivalve tissues of commercial bivalves from China and
two bivalve species from Oregon, USA [19,21], while fragments were the
majority of ingested microplastics in mussels from the Northern Ionian Sea [12].
Microplastics contamination also affected Asian green mussels, Perna viridis,
cultured in the coastal areas of Central Java, Indonesia [22].

Mussels and clams are among the favorable seafood products in Indonesia. The
coastal communities of Lamongan, East Java, for example, harvest, sell, and
consume marine bivalves. Ingesting contaminated seafoods could affect human
health [2,23]. In the present study, a survey on microplastic pollution in clams
from the coastal waters of Lamongan was conducted. The aim was to determine

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the abundance and type of microplastics present in wild clam tissues from
intertidal sediment of the area.

2 Materials and Methods

2.1 Study Site and Samples Preparation

Samples were collected from Lorena Beach (112.349439°, -6.867448°), Paciran
in June 2019, at the beginning of the dry season. The area is part of the coastal
areas of Lamongan, East Java with approximately more than 150,000 people
living in an area of only 120 km2 [24-25]. According to the Köppen-Geiger
climate classification, the study area has a tropical savannah climate with
temperature and precipitation averages at 27.4 °C and 1465 mm respectively [26].

In addition to its high population along the coastline, the area is situated between
the Sedayu Lawas River and the Ujung Pangkah River. Both rivers are part of the
delta of the Bengawan Solo River, the longest river on the Indonesian island of
Java (600 km in length). There are also many industrial and port activities around
the study area, which may increase the level of heavy metal pollution in its waters,
sediment, and cockles [27]. A previous study on microplastics in the area also
revealed high levels of microplastics, mainly fibers, in sediment from the Sedayu
Lawas River delta and across the coastal areas of Lamongan [7].

Figure 1 The location of sample collection, Lorena Beach, Lamongan, East Java,
Indonesia.

Clams were collected in the intertidal area at low tide, 1 km along the beach and
300 meter perpendicular to the coastline (Figure 1). A total of 80 wild clams were
sampled by wading and dredging the substrate at a maximum depth of 20 cm
below the surface of the sediment. The samples were separated according to their
species and were directly transferred to the laboratory and stored at -18 °C until

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further analysis. Prior to microplastics extraction, only clams with meat and
closed valves were considered for extraction, yielding 15, 8, and 15 clams from
the G. tumidum, A. antiquata, and V. philippinarum species, respectively.

2.2 Quality Control of Experiments

All liquids (natrium chloride solution, hydrogen peroxide and tap water) were
filtered prior to use with 0.45 µm filter paper to avoid sample contamination. All
glass bottles and other equipment used in the experiment were washed three times
with filtered tap water. Blank extraction procedures (n = 38 replicates) without
clam tissues and seawater were run to evaluate the background contamination.
There were 3 microfibers in all blank samples, representing 2.3% of all
microplastics extracted from the samples. Therefore, the blank contamination
was not subtracted from the results of this study. Furthermore, a probe and hot
needle test were performed when plastics were not able to be distinguished from
organic pieces.

2.3 Hydrogen Peroxide Treatment of Soft Tissue

The digestion method using hydrogen peroxide (H2O2) as oxidizing agent is
increasingly implemented. It is considered the most efficient method for the
destruction of organic matter for a wide variety of sample matrices [28]. H2O2 as
destruction agent has been used for the destruction of bivalve tissues [19,29,30],
as well commercial fish [31], anchovies [32], and even guano from Arctic seabird
colonies [33]. Furthermore, Al-Azzawi, et al. [28] compared preparation methods
for microplastic analysis and found that H2O2 digestion is the most effective in
terms of removal of organic matter from microplastic samples. In this study, the
extraction method of microplastics from clams was based on Li, et al. [29] with
minor modification. The shell weight, length, and width of each clam were first
recorded. The shells were then opened, and the soft tissue from each individual
clam was weighted and placed into a 750-mL glass bottle. In total, there were 15,
8, and 15 bottles for the clam species G. tumidum, A. antiquata, and V.
philippinarum, respectively. Approximately, 150 mL of 30% H2O2 was gently
poured into each bottle to digest the organic matter from each clam tissue. All
bottles were covered with foil to prevent airborne plastic contamination and
placed in an oscillation incubator at 65 °C at 80 rpm for 12 h. The samples were
then settled at room temperature for 24 to 36 h depending on the digestion status
of the clam tissue.

2.4 Floatation and Filtration with Natrium Chloride (Nacl)

Solution
The floatation method according to Li, et al. [29] was used to separate the
microplastics from the dissolved liquid from the soft tissues. Approximately 500

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 Muhammad Arif Asadi, et al..

mL of filtered NaCl solution was poured into each bottle and mixed thoroughly.
The solution was retained overnight and directly filtered over Whatman filter
paper (No. 42) using a vacuum system and was then placed into a petri dish with
a cover for further microplastics observation.

2.5 Microplastics Observation

The microplastics were observed and identified using an Olympus CX41
microscope. A visual assessment was applied to identify and classify the
microplastics particles according to their physical characteristics. The particles
were then classified as either fragment, film, fiber, or pellet according to Di &
Wang [34].

2.6 Statistical Analysis

Statistical analysis and graph design were performed using GraphPad Prism
9.0.0. The abundance of microplastics per species and the type of microplastics
per species were analyzed using two-way ANOVA. Šídák’s multiple
comparisons test was then used to compare the abundance of microplastics
between two species both in terms of items/individual and items/g. Also, the
microplastics types per species were compared using the same multiple
comparisons test.

3 Results and Discussion

3.1 Bivalve Measurements

The shell length and weight varied from 2.7 to 5.5 cm and 4.9 to 22.73 cm,
respectively, where G. tumidum had both the shortest shell length and the lowest
shell weight (3 ± 0.09 cm and 6.9 ± 0.33 g/individual, respectively). This species
also had the lowest soft tissue weight, 0.8 ± 0.04 g/individual (Table 1).

Table 1 Length, width, weight, and soft tissue weight of clams from coastal
waters of Lamongan, Indonesia.
Shell Soft tissue
Shell width Shell weight
Species Number length weight
(cm) (g/individual)
(cm) (g/individual)
Gafrarium
15 3 ± 0.09 2.4 ± 0.07 6.9 ± 0.33 0.8 ± 0.04
tumidum
Anadara
8 4.4 ± 0.44 10 10 3.1 ± 0.58
antiquata
Venerupis
15 3.7 ± 0.05 10 10 1.1 ± 0.05
philippinarum
Mean ± standard error (n = 8-15)

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 Microplastics in wild clams of Indonesia

3.1 Abundance of Microplastics in Bivalves

The number of total microplastics in the samples varied from 0 to 10
items/individual and 0 to 8.7 items/g (wet weight), while the total average of
microplastics concentration for all species was 3.5 ± 2.8 items/individual and
3.2 ± 2.52 items/g. Šídák’s multiple comparisons test of microplastics abundance
between two species showed significant differences (P < 0.001), where A.
antiquata had the lowest microplastics concentration (0.5 ± 0.37 items/g and
1.6 ± 1.51 items/individual), while, V. philippinarum had the highest
microplastics concentration (4.9 ± 2.80 items/g and 5.6 ± 3.22 items/individual)
(Figure 2).

Figure 2 Abundance of microplastics (items/g and items/individual) in clams

from the coastal waters of Lamongan, East Java, Indonesia.

3.2 Types of Microplastics in Bivalves

There were 130 microplastics extracted from the clams, where fibers accounted
for 80.77% of microplastics contamination. Meanwhile, both films and pellets
showed only a small percentage (0.77%) of microplastics ingestion. In A.
antiquata, 92.31% of microplastics were in the form of fibers and the remaining
microplastics were films. The highest share of fragments was in the G. tumidum
samples (22.86%), while no fragments were found in the A. antiquata samples
(Figure 3).

There were significant differences in the abundance of fibers and fragments in all
clam species (P < 0.001). The concentrations of both types of microplastics were
highest in V. philippinarum, 4.5 ± 2.59 items/individual and 1 ± 1
items/individual respectively, or 3.9 ± 2.39 items/g and 0.8 ± 0.78 items/g
respectively. Meanwhile, the lowest abundance of fibers was found in A.
antiquata, 1.5 ± 1.51 items/individual and 2.2 ± 1.56 items/g. Pellets and films
were only found in G. tumidum and A. antiquata respectively, with concentrations

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of 0.07 ± 0.25 items/individual and 0.09 ± 0.35 items/g for pellets and of
0.12 ± 0.35 items/individual and 0.37 ± 1.04 items/g for films (Figure 4).

Figure 3 Composition of microplastics types per clam species from the coastal
waters of Lamongan, East Java, Indonesia.

Figure 4 Composition of microplastics types per clam species from the coastal
waters of Lamongan, East Java, Indonesia.

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 Microplastics in wild clams of Indonesia

3.3 Size of Microplastics in Bivalves

The particle size for all microplastics, either in the form of fragments, pellets or
films, was less than 50 µm. Moreover, slightly more than half of the fibers (57%)
were in the size range of 3 to 50 µm, although 5.7% of fibers had a particle size
greater than 200 µm (Figure 5).

Figure 5 Particle size distribution of microplastics in the clams.

3.4 Discussion

3.4.1 Microplastics Ingestion by Clams

Microplastics ingestion in clams is widespread in the coastal waters of
Lamongan. Almost all samples (99.23%) were contaminated with microplastics,
ranging from 1 to 10 items in each clam. This indicates that the coastal waters
and sediment in the area are highly contaminated with microplastics particles. G.
tumidum, A. antiquata, V. philippinarum and most clam species are sediment
dwelling bivalves [35,36] and are filter feeders, which means that they feed by
straining suspended matter and food particles from the water in their surrounding
habitat [24,35]. The sediment of intertidal areas of Lamongan hold microplastics
at more than 200 items/kg [7]; therefore, the clams inevitably ingest microplastics
via the filter-feeder mechanism as microplastics within sediments are naturally
transported to bottom water [37].

Interestingly, despite its highest shell and soft tissue weight, A. antiquata ingested
far fewer microplastic particles than both G. tumidum and V. philippinarum, not
only by weight but also per individual. As the clams live in the same habitat, a
possible explanation for the differences in microplastics ingestion capacity are
species-specific traits. Bivalves, including clams, are selective particle feeders
and have the specific ability to select among microplastics particles and eject
microplastics in the form pseudo-feces and feces [38]. The difference in particle
selection depends on the gill architecture and the structure of the cirri [39].

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3.4.2 Types of Microplastics in The Clams

The overwhelming majority (>80%) of microplastics identified in the clam
samples were in the form of fibers. Fibers are the main type of microplastics, not
only in clams but also in their relatives, such as oysters, scallops, and mussels
[29,40-43]. Fibers have also been reported as the dominant type of microplastics
in other type of marine organisms, such as anchovies [32], zooplankton [44,45],
sea cucumbers [46], and sea turtles [47].

As filter feeders, bivalves, including clams, can reflect the variability of

microplastics types in water and sediment to an extent. In the sediment of the
intertidal area of Lamongan, fibers constituted 87% of microplastics particles [7];
therefore, the microplastics that were extracted from the clam samples were
mostly in the form of fibers. In mussels sampled from coastal waters of the United
Kingdom, fibers were also the main microplastic type identified in both seawater
and mussels, ranging from 50 to 90% [48]. However, no fibers exceeded a size
of 400 µm in the clams (Figure 5). Li, et al. [49] state that the limit of microfiber
internalization in clams is 500 µm.

Furthermore, in addition to the dominance of fibers in the coastal sediment of

Lamongan [7], the shape of microplastics may limit the internalization of
fragments in clams [50]. Microplastic fragments are derived from the breakdown
of larger plastic debris, which is mostly caused by physical, chemical, and
photodegradation processes over prolonged time scales [43,51]. Typically, the
shape of the fragments is often irregular, cracked, or semi-spherical [51]. The
internalization of microplastic in clams is also related to the softness and
smoothness of the microplastic materials; smoother fibers are easier to bend and
pass through the narrow openings of the clam’s gills [49].

3.4.3 Comparison of Microplastic Pollution in Bivalves

Microplastics ingestion in the clams collected from coastal waters of Lamongan
was widespread and showed a great variation among individuals and species
(Figure 2). Compared to the abundance reported in other studies (Table 2), G.
tumidum in this study had much lower microplastics content than in Pari Island,
Jakarta, Indonesia (13.44 to 17.33 items/individual, 9.1 items/g) [52]. In Jakarta
Bay, other bivalve species, Perna veridis, were also found to contain a high
number of microplastics, 5.39 to 39 items/g [53], while the total average of
microplastics concentration from all bivalve species from the Lamongan coast
was only 3.2 ± 2.52 items/g. Pari Island is located in Jakarta Bay. The sediment
of the bay holds at least 18,405 items/kg of microplastics [54], while the coastal
sediment of Lamongan sequestered only 206 items/kg [7].

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 Microplastics in wild clams of Indonesia

Although a considerably high number of microplastics, mainly fibers, was

observed in V. philippinarum from Baynes Sound, British Columbia, Canada
(0.07 to 5.47 items/g), V. philippinarum in the coastal area of Lamongan,
Indonesia had more ingested microplastic particles (Table 2). Clams from other
parts of the world also showed lower microplastic abundance; Donax faba from
the Gulf of Mannar, Cerastoderma edule from the Channel Coast, France, and
Siliqua patula from the Oregon coast, USA held a microplastics concentration of
0.6 to 1.3 items/g, 0.15 to 0.74 items/g, and 0.16 ± 0.02 items/g, respectively
[21,55,56].

Table 2 Comparison of microplastics contamination reported by this and other

studies.
Type of Abundance of
Species Location References
microplastics microplastics
Fiber (46%), 5.39-39
Perna veridis Jakarta Bay [53]
film (40%) items/g
Oregon coast, 0.16 ± 0.02
Siliqua patula Fiber (99%) [21]
USA items/g
Channel Coast, 0.15-0.74
Cerastoderma edule Fiber (50.2%) [55]
France items/g
Fiber (53%),
Gulf of Mannar, fragment 0.6-1.3
Donax cuneatus [55]
India (22%), film items/g
(21%)
Baynes Sound,
Venerupis 0.07-5.47
British Fiber (90%) [57]
philippinarum items/g
Columbia

4 Conclusions
Microplastics ingestion in clams from the coastal waters of Lamongan, Indonesia
is widespread, as almost all clam samples were contaminated with microplastics.
Therefore, this study provided further evidence that microplastics contamination
impacts marine organisms and can potentially be transferred to higher trophic
levels through the food chain. Furthermore, as the most ubiquitous microplastics
type ingested in the clams, fibers are an underestimated threat to aquatic
environments. Policy and regulations need to be implemented to reduce the
release of synthetic fibers in the marine environment.

Acknowledgement
This study was part of the Hibah Penelitian Pemula (HPP) research scheme of
University of Brawijaya. The authors would like to thank the dean of the Faculty
of Fisheries and Marine Science and the rector of the University of Brawijaya for
their support of the research and this publication.

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 Muhammad Arif Asadi, et al..

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