Isolation and identification of microplastic on the cockleshell of blood clam (Anadara Granosa Linn)

This study aims to figure out the occurrence of microplastics in cockleshells of Anadara granosa Liin from the five beaches. The dried cockleshells were immersed in NaCl for the partition step in view of the particular gravity of the microplastic and categorized according to shape under a stereo microscope. In all samples, were consistently found microplastics. The beach that has the largest abundance of microplastics is the Kenjeran Beach Surabaya in East Java Province. The shape of the abundance of microplastics found in this research were fragments, film, foams, crystals, granules, pellets, filaments, and yarn. The average microplastic abundance was 171 pieces per 75 grams of cockleshell dry weight. Among the identified microplastics, fragment (55.33%) was the most frequent type. The occurrence and distribution of microplastics in cockleshells of specimens from the studied region provided information for evaluating the risks of microplastics.


Introduction
The microplastic has been reported in the surface waters of every ocean [1], in sediments, mangroves, intertidal areas, coastal areas and sands of the deep sea [2], as well as bivalves group organisms [3].
Channel taking care of marine life forms, for example, shellfishes are inclined to ingest microplastics because of taking care of and gill ventilation systems [3].Ingestion of plastic waste was also found in shellfish [4].Ingested microplastics were accounted for to be caught and amassed in shellfish tissue.Microplastics have also been found to move through food webs in water [4].
Studies have shown that microplastics are eaten by both developed and wild mussels, in this manner influencing their development and slowing down the section of food through the stomach [5] and [6].Microplastics in the ocean also increase the adsorption capacity of heavy metals [7], which might affect human well-being and marine nature.System functionality raises risk.Microplastics can possibly become transporters of tenacious natural contaminations in marine fish and shellfish and, hence, in people [8].The potential health effects of microplastic chemical toxicity depend on the size, shape, type of polymer, density, chemical composition, and absorption kinetics of microplastics [9]and [10].
The investigation of microplastic defilement in fish for human utilization is significant in forestalling and moderating the potential well-being influences for the purchaser's local area.Also, microplastic research gives data to the hydroponics local area and neighbourhood state-run These animals ingest microplastics directly or through predation of organisms at lower levels of the food chain [11].Based on the research of [12] on shellfish taken from the free seas of China, they stated that microplastics were found in shellfish with an average number of up to 4.6 plastic particles per sample.The small size of microplastics is considered as food by marine biota, resulting in the migration of microplastics from low trophic to high trophic [13] and [12].
The high production of clam is followed by high shell waste [14].The utilization of waste shells containing calcium and phosphorus minerals began to be carried out.Various uses are made not only to make wall decorations but also for animal feed, cosmetic mixtures, calcium fortification in food [15], abrasives in toothpaste and food preservatives.
This study was to visually identify and analyze the presence of microplastics in the cockleshell of blood clams (Anadara Granosa Liin) taken in 5 areas and compare their abundance.This study was conducted at Sidoarjo Regency, Gresik Regency, Pasuruan and Surabaya.These beaches include areas with high anthropogenic activity, originating from residential areas, plantations, fishing industry, aquaculture and tourism sectors around the river mouth to the coast.

Study Location
Samples of blood clam cockleshells (Anadara Granosa Liin) were taken at 5 locations: Kenjeran Beach Surabaya City; Gisik Cemandi Beach Sidoarjo Regency; Lekok Beach Pasuruan Regency; Mengare Beach Gresik Regency, and Lingkar Timur Beach Sidoarjo Regency, in East Java Province, Indonesia (Figure 1).The equipment used to reduce shells to small fragments was a drying oven, a crusher and a mesh sieve.Equipment for observation was a stereo microscope, a 500 ml glass beaker for immersing cockleshells, a funnel, a container to hold the filtrate, a sample bottle, a petri dish, and filter paper.

Procedure for making cockleshell flour
The cockleshell samples were dried, then crushed and sieved using a 200 mesh sieve [16].The sample preparation process, visual identification and analysis of samples at the Ecoton Surabaya Laboratory.

Sample Analysis Method and Microplastic Types
The method of analyzing microplastics in the laboratory referred to the research of [17] [18].The destruction of organic matter using a solution of 30% Hydrogen Peroxide and 0.05 M Fe (II) of 20 ml each [19].The dried example was then added with 300 ml of immersed NaCl for the partition step in view of the particular gravity of the microplastic [20].
Visual perception of microplastics on cockleshells was done by pouring 3-5 ml of test arrangement into a petri dish and utilizing a sound system magnifying instrument.ID of microplastics was done by utilizing the visual perception strategy by taking a gander at the state of the microplastic particles [21].The microplastics acquired were then included in the number.It distinguished the sorts of microplastics as pieces, granules, films, pellets, fibres and froths [22].

Figure 2. Based On Visual Observations And The Use Of A Microscope, Total Number Of
Microplastics In Five Sampling Locations The fibre was identified by its elongated shape with the same thickness; fragments are particles with an irregular shape and can not be crushed using tweezers, and can be in a variety with a stiff, thick texture, with sharp and bent edges; thin sheet films which are usually crushed from plastic bags or food packaging, are irregular in shape, thin and flexible and usually transparent; pellets come from the plastics industry.They are irregular in shape, round, and generally larger, with a width of about 5 mm.Granules are generally flat on one side and can be of a variety of tones, have a standard round shape and are usually smaller in size, around 1 mm in diameter, and appear in regular tones of white, cream, brown, etc.The most abundant kind of plastic particle is fibre.They can be short or long, with various thicknesses and varieties, and the froth most frequently comes from enormous styrofoam particles.They are delicate, sporadic in shape, and in colour variety [22].

3.1.Total Abundance of Microplastics in Five Sampling Locations Based on Visual Observations
Using a Microscope The five coastal areas are ship traffic lanes.Most of the microplastics found came from fishing boats and fishing gear.The location distance between mangroves, fish auction sites and ponds is not far, so the types of microplastics found are relatively the same.Meanwhile, the highest anthropogenic activities were at fish and mangrove auction sites.Mangroves also have a high occurrence because waste can be trapped in the mangrove roots.In Figure 3 and Figure 4, compared to the fragments and fibres in almost all locations, there is also a very small amount of film-type microplastics.
The five coastal areas are ship traffic lanes.Most of the microplastics found came from fishing boats and fishing gear.The location distance between mangroves, fish auction sites and ponds is not far, so the types of microplastics found are relatively the same.Meanwhile, the highest anthropogenic activities were at fish and mangrove auction sites.Mangroves also have a high abundance because waste can be trapped in the mangrove roots.In comparison to fragments and fibres at almost all locations, Figure 3 also indicates that the types of film microplastics are very small.
Wellsprings of microplastics are the consequence of the fracture of bigger plastics conveyed by waterways, spillover, tides, and winds and afterwards moved by sources from the ocean, which incorporate fishing gear, development hardware and apparel filaments from homegrown wastewater steps [24].Notwithstanding the contribution from land, the high grouping of microplastics is prompted by passing boats that make a significant commitment to microplastic contamination [25].

Figure 3. Shape and Total Abundance Microplastics in Five Sampling Locations Based on Visual
Observations Using a Microscope The same thing was found at the research location.Microplastics with lower thickness will generally drift.Film-type microplastics have a lower thickness, so they will quite often drift in the water segment and are more straightforward to ship [26].At the location of the fish auction, mangroves and ponds are strongly influenced by tides, so the film particles in these two areas are found very little because they are easier to use drifting.

Shape and Total Abundance Microplastics in Five Sampling Locations Based on Visual Observations Using a Microscope
The kind of microplastic that has the most elevated typical overflow is a piece, and this connects with a similar poison source.The wellspring of microplastic contamination comes from anthropogenic exercises, for example, family waste and fishing exercises, which are remembered to contribute to the biggest microplastic sections.Microplastic contamination also comes from rivers and seas that enter the coast.
[27] found that microplastic in the form of fibre dominates Surabaya marine sediments by 57%, in the form of filaments by 36%, and fragments by 7% in Surabaya marine sediments.Many types of fibre are found because they come from nets used by fishermen [28].[27] reported that the FTIR test results in the Jagir -Wonorejo Estuary of Surabaya had identified polymer compounds (microplastics) in the form of Polyester.
Polyester is the most abundant type due to its density being almost as dense as seawater.On the other hand, the density of Low-Density Polyethylene and Polypropylene was lower than that of seawater.However, polypropylene and Low-Density Polyethylene particles were also found in aquatic sediments.The cause could be biofouling, and biofouling can deposit particles to the seabed because it can increase the density and weight of microplastics.The patterns of abundance of microplastics found between the waters in this research were fragments more of film, more foams, more of crystals, more of granules, more pellets, more filaments, and more yarn.
Most forms of fibre are produced from textiles and fishing gear [28].In addition, many laundry businesses are growing in metropolitan cities that do not have operating permits and wastewater treatment facilities [27].Laundry liquid waste is generally disposed of directly into a drainage channel that empties into a river.Past investigations have announced that the washing system can deliver 1900 -700000 Fiber/6kg textures into the climate, and fibre poisons additionally come from homegrown waste because of material washing exercises and the utilization of care items [29].The deposition process and the speed of fibre deposition make the particles settle more easily than other forms of particles in nature.
Sources of microplastic fragments obtained came from bottles, plastic bags and pieces of pipe.In addition to fragments, microplastic fibre and film types were also found in this area.Wellsprings of fibre-type microplastics are remembered to come from manufactured textures, squandered from fishing boats and fishing stuff like fishing nets and fishing lines.The wellspring of film-type microplastics comes from food bundling.The majority of the wellsprings of these microplastics come from human exercises and are impacted by flows and tides.Most forms of fibre are produced from textiles and fishing gear [28].In addition, many laundry businesses are growing in metropolitan cities that do not have operating permits and wastewater treatment facilities [27].Laundry liquid waste is generally disposed of directly into a drainage channel that empties into a river.Past investigations have announced that the washing system can deliver 1900 -700000 Fiber/6kg textures into the climate, and fibre poisons additionally come from homegrown waste because of material washing exercises and the utilization of care items [29].The deposition process and the speed of fibre deposition make the particles settle more easily than other forms of particles in nature.

Microplastic Visual Data using a microscope visual
Sources of microplastic fragments obtained came from bottles, plastic bags and pieces of pipe.In addition to fragments, microplastic fibre and film types were also found in this area.Wellsprings of fibre-type microplastics are remembered to come from manufactured textures, squandered from fishing boats and fishing stuff like fishing nets and fishing lines.The wellspring of film-type microplastics comes from food bundling.The majority of the wellsprings of these microplastics come from human exercises and are impacted by flows and tides.
From Figure 4 to Figure 8, microplastics in the form of films are the most commonly found microplastics in the Brantas River in Surabaya, Indonesia [31].Based on Qing's research on China's Pearl River, the results of this study are similar [32].Based on the shape of this microplastic, it can be estimated where it came from.The shape of this plastic is similar to the shape of plastic packaging.Past examinations led in Singapore Mangroves and the Bay of Mexico assessed that the type of microplastic film type comes from plastic bundling and pop packs.This condition is because of the popularity of plastic bundling.

Conclusion
The average abundance of microplastics in clamshells (Anadara Granosa Liin) from 5 (five) water bodies was 171 particles/75 g of sample.It is the type of microplastics found in water that are fragments, films, granulates, granules, foams, fillers, crystals and threads.The highest frequency of microplastic fragments was found at all sites.This is because microplastic pollution comes more from household waste and debris from human activities.Many types of research related to microplastics in clam meat have been carried out, but are the microplastics found in clam meat also in clamshells?This is due to the increasing use of cockleshells for animal feed that will be consumed by humans.Hence, it is important to explore the microplastic content in cockleshells, particularly blood cockleshells, which are the most elevated maker in fishery creation.The study is part of a risk analysis of microplastics in seafood.