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Microplastic Ingestion by Crabs

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6th
PROCEEDING OF
CIVIL
ENGINEERING
The 6th Proceeding of Civil Engineering

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Edited by:

Dr. Dayang Zulaika Abang Hasbollah

Dr. Ain Naadia Mazlan
Dr. Nur Hafizah Abd Khalid
Dr. Mohd. Ridza Mohd. Haniffah
Dr. Libriati Zardasti
Dr. Kogila Vani Annammala
Dr. Mohamed Zuhaili Mohamed Najib
Dr. Muhammad Naqiuddin Mohd Warid
Dr. Erwan Hafizi Kasiman
 - 235 -

PART 2:
WATER AND
ENVIRONMENTAL
ENGINEERING

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Microplastic Ingestion by Crabs
Yau Zi Yi1, Shamila Azman1*, Audrey Primus1, Mohd
Ismid Mohd Said1, Muzaffar Zainal Abideen1
1
School of Civil Engineering, Faculty of Engineering, Universiti
Teknologi Malaysia, Malaysia
*
shamila@[Link]

ABSTRACT. Microplastics are anthropogenic contaminants with a diameter < 5

mm and are readily present in various morphology. The detrimental effects
resulted from microplastic pollution are becoming a growing concern due to their
omnipresence in the marine environment. It is perceived that the ecotoxicological
effects of microplastics retained in the edible marine species will further impact
food safety. Crab species are susceptible to microplastic ingestion due to their
natural feeding behaviour coupled with burrowing activities. This paper reviews
the current knowledge and research findings on the ingestion of microplastics by
various crab species. The review also covers the adopted methodology for
digestion of biological tissues and microplastic identification and impacts of
microplastics to the ecosystem as well as the implications for food security
concerns. Current knowledge gaps and future perspectives are also discussed as
well in this paper.

Keywords: Microplastics; Crab; Ingestion; Food Security.

INTRODUCTION

The versatility, durability, corrosion-resistant ability and inexpensive

manufacturing cost of plastics have made them preferable in manufacturing a
wide range of synthetic products. However, the intensive application of plastics
and their derivatives has led to the accumulation of plastic wastes in terrestrial
land and marine environment, thereby triggering ubiquitous plastic pollutions all
over the world [1]. Series of weathering and degradation processes result in the
fragmentation of large plastic debris into micro-sized fragments. The term
‘microplastics (MPs) is used to define the plastic fragments with a diameter less
than 5 mm [2]. The dissemination of microplastics over the global marine
ecosystems have progressively received wide concern regarding their possible
negative effects on aquatic organisms and natural ecosystems. The microplastic
particles are readily ingested by the aquatic species due to their relatively small
size and resemblance to natural prey [3]. Several scientific studies reported the
effects of unintentional ingestion of microplastics on influencing the physical
health of aquatic species.
Fish and other seafood (bivalves, crustaceans and shellfishes) serve as the
essential sources of protein for humans. However, seafood consumers are
susceptible to microplastic exposure when they consume the superior predator
marine species which are perceived to bear with the highest microplastics

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concentration due to the effects of bioaccumulation and biomagnification [4]. It
is suspected that crab species are highly vulnerable to microplastics
contamination as various edible crustacean species (prawns, lobsters, etc.) had
been proven for microplastics ingestion.
In this study, the main focus is drawn on crab species. Various crab species
serve as a source of income for fisheries and aquaculture, whereas they also play
an important role in the marine ecosystems as natural predators, prey or even
detritus feeders. For the consideration of the ecological and commercial
importance of crab species, research on microplastics contamination in this
aquatic species should be emphasized.

Problem Statement
It is beyond the shadow of a doubt that numerous studies had reviewed the
abundance, characteristics of distribution, bioavailability and ecotoxicology of
microplastics on the freshwater and marine aquatic species. However, the
available research on microplastics contamination in crab species is still limited
compared to other aquatic species such as fishes and bivalves.
The involuntary ingestion of microplastics pose risk to the marine
organisms and the dietary intake of seafood result in the trophic transfer of
microplastics into consumers. The retention of chemicals-containing
microplastics in the edible tissues of aquatic species is a cause of concern since
the prospective effects of microplastics abundance in the human body are not yet
comprehensively studied. Besides, the standardized database on the techniques
for microplastic identification is also scarce. To date, there are various
methodologies adopted by researchers to determine the microplastic
contamination in aquatic biota. This disparity often results in a contradiction in
making a comparison of data.

Objectives
The main purpose of this study is to investigate the presence of
microplastics in crab species. The objectives of this study are:
1. To summarize the presence and distribution of microplastics in different
crab species across different continents;
2. To discuss the method of digestion for biological tissues and
microplastic identification;
3. To access effects of microplastic retention in crab species and
implication with food safety.

Scope of Study
The study focuses on the microplastic ingestion by crab species, in which
various aquatic species have been used as experimental samples for microplastic
contamination such as bivalves, fishes and zooplankton. The literature review
does not limit to specific regions or continents to provide an overview of
microplastic ingestion by crabs originated from various locations all around the
world. Detailed review analysis will be conducted by extracting pivotal data from
all the achieved articles to compare the advantages and limitations of each

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microplastic digestion and identification method and summarize the perceived
effects of microplastic ingestion on crabs and the implications on human health.

LITERATURE REVIEW

Plastics are manufactured into diverse products due to their outstanding

features such as moldability, lightweight, durability, corrosion-resistant and high
electrical insulation [5]. However, extensive application of plastic and its
derivatives has triggered the accumulation of anthropogenic plastic debris in our
natural environment due to their chemical inertness and low rate of
biodegradation. Microplastics are used to define plastic fragments with a diameter
less than 5 mm [2]. In general, microplastics are classified based on their source
of origin: Primary microplastics are micro-size plastic particulates such as
microbeads originated from personal care products and industrial scrubbers;
Secondary microplastics are readily formed from the breakdown of large-sized
plastic debris into small fragments [6]. Due to the coupling effects of wind action,
stormwater runoff and the nature of buoyancy, microplastics debris are globally
omnipresent as they travel over vast distances within the marine environment.
The presence and abundance of microplastics have been identified in sediments,
freshwater, shorelines, seabed, polar region and aquatic biota that comprising of
edible species [7][8][9][10].
Microplastics ingestion have been attested in a wide range of organisms
from all trophic levels from pelagic zooplanktons to benthic mammals. It is
perceived that the ecotoxicological effects of microplastics resulted from
properties enhancement additives potentially retained in their bodies. The type
and amount of microplastics ingested by the marine organisms vary with their
foraging behaviours, feeding strategy and choice of dietary intake. Filter feeding
species such as bivalves, zooplankton and copepods are more likely to ingest low
density floating particles on the sea surface whereas high-density plastics, such
as polyvinyl chloride (PVC) which tend to settle as sediments, commonly being
ingested by deposit feeder species like sea cucumbers, crustaceans and flounders
[11].
Before the extraction of microplastics, digestion has to be performed to
dissolve the biological matrices in the sample without destructing the plastic
polymers. Several digestion methods are being introduced in various research
papers such as using acids, alkaline, oxidizing agent or enzymatic methods [12].
There are three common analytical methods to identify polymer constituents of
microplastics, which are visual inspection, chromatography/ thermal analysis and
spectroscopy techniques [13]. The efficiency and limitations of each digestion
and identification method will be assessed in this paper.
Crab species are readily exposed to microplastics loading due to their
feeding behaviour coupled with burrowing activities. Hence, the likelihood of
microplastics pollution reported in these species is either via involuntary ingestion
of sediments or through the consumption of microplastic-contaminated prey [14].
The retention of microplastic debris in the edible tissues of crabs suggests the
further trophic transfer in the food web, thereby emerging the food safety concern.

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METHODOLOGY

Sourcing Reference Material

To achieve the objectives of this study, literature retrieval has been
comprehensively conducted through various resource websites such as
ScienceDirect, Scopus, ResearchGate, AsloPubs, Pubmed and Academia. The
snowballing method is applied in collecting the data, where derived literature was
accessed in the references of the reviewed journals. The search terms include
“microplastics,” “ingestion,” “crabs,” and “aquatic biota”. All references
achieved are published between the year 2000 to 2021.

Quality Control of Data

All the achieved literature was preliminarily reviewed by subject
disciplines and titles before the full-text screening. The relevant data was
conscientiously extracted. To ensure the quality of the collected data for the study,
the following criteria were assessed: (1) experimental replication was conducted
for statistical analysis; and (2) precautions or any relevant contamination controls
were implemented in the study protocol such as negative controls in toxicity
testing, air-borne contamination controls throughout laboratory procedures with
simultaneous processing of procedural blanks. If any of these quality control
parameters were not satisfied, the literature was excluded from this paper. In
addition, the selected literature was cross-referenced with existing data showing
crab species with global commercial importance to aquaculture and fisheries. A
total of 50 numbers of literature were retrieved to ascertain the data presented in
this review.

RESULTS AND DISCUSSION

Table 1 shows the summary of retrieved literature on microplastic

ingestion by crabs worldwide, where the extracted data is arranged accordingly
from latest to oldest. Notably, flower crabs have been selected as sampling species
for microplastic ingestion in several studies due to their commercial value to be
served as a popular delicacy, particularly within the Southeast Asia region. As
stated in most of the review studies in Table 1, fibres seem to be the predominant
type of microplastics that are readily ingested by crabs. These fibres are most
probably originated from synthetic textiles and fishing nets. Due to the greater
density than water (range from 1.09 to 1.20 g cm −3 or  > 1.35 g cm−3), these fibres
would precipitate and settle down in the underlying sediments where the deposit-
feeding crab species feed. The magnitude of ecotoxicological effects of
microplastics on the aquatic species varies depending on the shape of
microplastics, where fibre will lead to more severe toxicity as compared to other
shapes like fragments and beads [15].

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Table 1: Literature records of the microplastics ingestion by crabs from various
continents
Location Crab Species Type of Detection Concentration Characteristics Reference
Organ Method of MPs of MPs

Fishing Flower crab Stomach FTIR 43 ± 7 grams Mostly [16]

harbours of (Portunus tissues spectrometry per flower crab fragments,
Kerala, India pelagicus) transparent and
white, size range
from 100 to 200
μm
Yellow Sea, Gazami crab Gills and μ‐FTIR 5 ± 4 items per Mostly fibres, [17]
East China Sea (Portunus gut spectrometry individual black-grey and
trituberculatus) blue-green with
sizes ranged
from 19.97 to
4976.22 μm

Port Blair Bay, Flower crab Gastrointe ATR-FTIR 5 number of Mostly fragments [18]
Andaman (Portunus stinal tract spectroscopy items per and size less than
Islands, India pelagicus) individual 500μm
Thames European green Gills and 8.78 ± 0.83 Mostly fibres, [19]
Estuary, crab gastrointes FTIR number of clear in colour
the Kent / Essex (Carcinus tinal tract spectroscopy items per and mean length
Strait, Great maenas) individual of 2.1mm
Britain
Corpus Christi Chesapeake blue Stomach μ‐FTIR 0.87 number of Mostly fibres, [20]
Bay, Texas crab tissues spectrometry items per crab transparent,
Coast, Gulf of (Callinectes white and yellow
Mexico sapidus)
Persian Gulf, Flower crab Gills and SEM-EDX 0.863 items per Mostly fibres, [43]
Iran (Portunus muscle analysis muscle black with size
armatus) tissues (Scanning less than 5mm
electron
microscopy
coupled with an
energy dispersive
X-ray (EDX)
unit.
California Pacific mole crab Digestive ATR-FTIR 0.65 ± 1.64 ite Mostly fibres [21]
Coast, United (Emerita analoga tract spectroscopy ms per
States ) individual
The Celtic Sea, European spider Stomach FTIR 1.39 ± 0.79 Mostly fibres [22]
south coast of crab tissues spectrometry number of
Ireland (Maja squinado) items per
bordered by the spider crab
English
Channel and
the Atlantic
Ocean
Bering-Chukchi Snow crab Soft tissue FTIR 0.04–1.67 Mostly fibres, [23]
seas continental (Chionoecetes and gills spectrometry number of red with mean
shelf, Artic opilio) items per sizes
region individual 1.45 ± 0.13 mm

Vistula Lagoon, Chinese mitten Stomach Microscopy 18 out of 208 Mostly fibres, [24]
Russia crab tissues individuals clear in colour.
(Eriocheir recorded
sinensis)

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microplastic
contamination

Microplastic Ingestion by Crabs

The colour of ingested microplastics varies according to species, which
are likely correlated with the species' location and feeding habits. According to
Zhang et al. [17], the size and colour of the particles have direct implications on
the microplastic ingestion by crabs, which are commonly known as visual
predators. The microplastic debris might have an identical size as sand, and their
colour might be similar to the colour of potential prey, which may cause mistakes
in dietary intake for a visual predator. The study on Chinese mitten crab
(Eriocheir sinensis) by Wójcik-Fudalewska et al. [24] elucidated the majority of
microplastics detected from these species consist of clear balled-fibres. The
outcome of the study matches the prevalence of fisheries in the sampling location,
where the fibres are perceived to be originated from fishing nets, ropes and gears.
Based on the reviewed literature, the common types of polymers reported
in microplastic ingestion by crab species are as the following: polyester (PET),
polystyrene (PS), polyethene (PE), polypropylene (PP) and polyamide (PA).
Polyester (PE) appeared to be the most abundantly ingested type of polymer by
crabs due to their prevalence as synthetic fibres in modern textiles. Having a
greater density than water (range from 1.09 to 1.20 g cm −3 or  > 1.35 g cm−3), the
polyester fibres are readily precipitate and settle down in the underlying
sediments where the benthic crab species feed. Although the identified polymers
did not allow tracing of their origin, polyethene (PE) and polypropylene (PP) are
generally originated from product packaging, medicine bottles, automobile parts,
kitchenware and shampoo containers [25]. Polystyrene (PS) is widely used in
laboratory apparatus, disposable plastic cutleries and fishing buoys, whereas
polyamide (PA) is commonly used as synthetic fibre in making robust fishing nets
and gears [26].
Most of the studies focused on the stomach tissues and gastrointestinal
tract on the detection of microplastics contamination. Some researchers reviewed
the microplastics exposure on other susceptible viscera such as gills. It is
suspected the likelihood of entanglement of microplastics, particularly in fibre
form, within hairs or folds on the internal organ’s surface when crab inspires by
ventilating water from the surrounding into the gills [27]. Studies had shown the
retention of microplastics in the gills of various marine organisms, in which the
abundance is comparable to those in the guts of organisms [28]. In addition, the
translocation of microplastics from the gut to other tissues in crabs is also
demonstrated by numerous laboratory researches. Figure 1 shows the uptake and
retention of microplastics within the internal organs of the European green crab,
Carcinus maenas according to the study by Watts et al. [27].

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Figure 1: Uptake and retention of microplastics within the body tissues of

European green crab, Carcinus maenas (Adapted from Watts et al. [27])

Digestion Methods of Biological Tissues

Before the extraction of microplastics, a digestion procedure has to be
performed to dissolve the complex matrices in the biological sample without
destructing the plastic polymers. To date, several digestion methods are being
introduced in various research papers such as using acids, alkaline, oxidizing
agent or enzymatic methods [12]. However, it is indispensably vital for
researchers to take appropriate precautions while applying these chemical
digestion methods, since certain chemicals will destroy the microplastics and
thereby contributing to the microplastics concentration underestimation [29].
Dehaut et al. [30] investigated the efficacy of biological tissues digestion based
on six protocols using different digestive chemicals. The authors suggested the
digestion method using 10% KOH solution (coupled with 24 hours incubation at
60 °C) to be the most compromising practice as it led to efficient digestion of
biological tissues (from mussels, crabs and fishes) without any significant
degradation noticed on all the tested plastic polymers and met the
recommendations by Marine Strategy Framework Directive (time effective and
low procedural cost).
There are certain limitations for the other digestion method: protocols
using nitric acid (HNO3) will be led to the degradation of certain types of
polyamide [31]; enzymatic approaches are expensive and time-consuming,
particularly for the large-sized species, despite its high digestibility reported [32];
application of oxidizing solution such as sodium hydroxide (NaOH) is less
effective as incomplete digestion reported on mussel tissues as well as
degradation on the structural integrity on plastics with types of cellulose acetate
(CA), polycarbonate (PC) and polyethylene terephthalate (PET) [30].
Methodology, terms and measuring units should be standardized for the
extraction of microplastics in different aquatic biota to minimize contradiction in
making a comparison.

Microplastic Identification Methods

There are three common analytical methods to identify polymer
constituents of microplastics, which are visual inspection, chromatography/
thermal analysis and spectroscopy techniques. Visual identification of
microplastics using a microscope is usually not recommended as it is prone to
have erroneous analysis despite its uncomplicatedness [33]. As stated in Table 1,

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most of the studies applied FTIR spectroscopy for microplastic identification in
the biological samples. FTIR spectroscopy has been experimentally proved as a
simple, efficient, inexpensive and non-destructive method for identifying the
characteristics of plastic polymers [34]. FTIR spectroscopy is advantageous in
providing information on the specific chemical bonds of particles, in which
carbon-based polymers can be easily identified. Different bond compositions will
produce unique spectra that distinguish plastics from other organic and inorganic
particles. The application of this method enables the identification of specific
types of polymer-based on the well-established polymer spectrum library.
However, there is a high possibility of missing tiny and transparent plastic
particles to be detected as microplastics with lengths less than 50 μm may require
trials to obtain a clear spectrum for precise identification.
Raman stereoscopy is another chemical characterization method used for
microplastic identification. This method will produce a distinctive spectrum for
each type of polymer through the projection of laser beam on the suspected items,
which yielding different frequencies of back-scattered light that are contingent on
the molecular structure and atoms present [35]. Besides identifying plastics,
Raman spectroscopy is also capable of providing polymer composition profiles
of each sample similar to FTIR spectroscopy. For the purpose to obtain a better
spectrum from particles less than 20 μm in size, Raman spectroscopy (in which a
laser beam with a smaller diameter is applied) is recommended to be used.
However, this method is reported to be sensitive to properties enhancement
additives in microplastics, which interfere with the polymer type identification
[34].
Thermal analysis is a technique that measures changes in the physical and
chemical properties of polymers based on their thermal stability. Differential
Scanning Calorimetry (DSC) is one of the thermal analytical methods that
investigates the thermal properties of polymeric materials. An abundance of
reference materials is required for polymer types identification as each plastic
materials demonstrate distinct characteristics in DSC. Pyro-GC-MS is a
combination of thermal analysis and gas chromatography-mass spectrometry
(GC-MS) that analyses decomposed gas thermally from polymers. This method
is capable of analyzing the chemical additives in microplastics simultaneously.
However, thermal analysis is seldomly used in studies is due to its
destructiveness, which interferes with the subsequent analysis of microplastic
samples. In addition, thermal analysis is considerably time-consuming in data
processing compared to the spectroscopy methods [34].

Effects of Microplastic Retention on Crabs

Various health impacts of microplastic retention on crabs have been
ascertained in recent research papers. According to Jeong et al. [36],
developmental delays, delayed moulting and reduced fertility are reported as the
physiological effects of microplastics retention on crustaceans. Furthermore,
microplastic ingestion showed the increased mortality of nauplii and copepodites
(crustacean larvae), thereby reducing the survival rate of juvenile crustaceans
[37]. The presence of chemical-additives-containing microplastics in the cellular
tissues can negatively affect the immune system functionality of crabs, in which
reduction in oxygen consumption and sodium (Na +) concentration, as well as the

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increase of hemocyanin in the haemolymph, had been proven inshore crabs [38].
However, the current knowledge on the mechanisms by which microplastics exert
these effects on crustaceans remains scarce despite the adverse effects of
microplastic ingestion that have been pervasively reported. It may be stated
without fear of contradiction that crabs play an indispensable vital role in the food
web of the coastal and marine ecosystems as natural predators, prey or even
detritus feeders. The stability of the crab community will significantly influence
the energy flow and material circulation in marine ecosystems. Hence, further
comprehensive studies and data extrapolation about microplastic contamination
on crabs are necessary.

Potential Food Safety Concern

Humans, as the top superior predator in the food chain, are susceptible to
be exposed to marine pollutants due to the processes of bioaccumulation and
biomagnification. Akhbarizadeh et al. [43] evaluated the effects of
biomagnification of microplastics through the seafood web by investigating the
relationship between weight, trophic level (TL) and microplastics content in the
samples’ muscle. The statistical results of the biomagnification factor (BMF) and
trophic magnification factor (TMF) calculation suggested that microplastics
concentration did not biomagnify in edible tissues of the marine food web in the
Persian Gulf, Iran. To date, the effects of microplastics biomagnification for
higher predators like humans are still poorly understood and detailed investigation
is demanded to ascertain the trophic transfer of microplastics in edible aquatic
organisms from different continents.
Microplastics are globally recognised as highly bioavailable contaminants
to marine organisms and their consumers [39]. The detrimental effects of
microplastics translocation have triggered an alarming concern on food safety and
human health among the scientific community. The overall and long-term impact
of microplastics retention in the human body remains unclear but research studies
have demonstrated oxidative stress development, tissue inflammation,
cytotoxicity and alteration in human sex steroids metabolism as a result of
microplastics exposure [40]. A study by Dong et al. [41] attested the adverse
effects of polystyrene exposure on human pulmonary cells. The researchers
employed the human lung epithelial BEAS-2B cells to investigate the implication
of microplastic exposure with pulmonary toxicity. Assay results revealed that
polystyrene exposure (concentration range from 1–1000 μg/cm2) for 24 and 48
hours significantly reduces the viability of the BEAS-2B cells by inducing
cytotoxic and inflammatory effects through the formation of reactive oxygen
species. In addition, Deng et al. [42] studied the effects of chronic microplastic
retention in the liver of house mice (Mus musculus) and reported several
detrimental effects, such as energy and lipid metabolism disturbance and
neurotoxic responses disorder. The concern is emerged for the effect of cellular
toxicity by microplastics uptake upon human liver cells. Although most of the
laboratory studies tend to apply a considerable microplastics concentration to
stimulate the biological response, the results from these short-term studies can
serve as indicators to evaluate the possible risks triggered by the prolonged
exposure to microplastics. Long-term exposure studies using environmentally
relevant microplastics concentration and having a morphological resemblance to

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those obtained from field studies should be conducted for a better comprehension
on the health implications of microplastics in the human body.

CONCLUSION

The accumulation of plastic debris in the marine environment has triggered

a growing concern about microplastic contamination in the aquatic biota.
Microplastic ingestion has been reported in various aquatic species, particularly
fishes, bivalves and pelagic planktons. However, studies on crab species are still
limited in different continents around the world. The ingestion of microplastics
by crabs might negatively affect their biological and physiological health.
Moreover, involuntary ingestion of microplastics might occur when humans
consume crabs. Thus, the potential risk assessments of microplastic exposure
upon human health are highly demanded, owing to the anticipated increase of
microplastic abundance in the natural environments. The findings that can be
drawn out from this study are as the following:
1. Fibres predominate as the most ingested types of microplastics by crabs as
they tend to settle down in the underlying sediment where the benthic crabs
feed.
2. The sizes and colours of microplastics ingested by crabs are correlated
with their feeding strategy and areas of habitation.
3. Microplastics are found abundantly in crabs’ stomach tissues and
gastrointestinal tract, but the translocation of microplastic into other
internal organs are attested.
4. Digestion method using 10% KOH solution with incubation at 60 °C for
24 hours appears to be the most compromising protocol with relatively
high efficiency of biological tissues without significant degradation of
plastic polymers.
5. FTIR spectroscopy is preferable by worldwide researchers invalidating the
microplastics polymer types in environmental samples due to their
uncomplicatedness and non-destructiveness.
Further research and assessments are required to address the following
uncertainties including the fate of microplastics within the internal organs of
aquatic organisms by residence time evaluation; the possibility of
biomagnification via trophic transfer of microplastics; the long-term effects of
microplastic intake on human health.

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