Effects of Different Polystyrene Concentrations on the Bioaccumulation of As, Cd, and Pb in Ruditapes Philippinarum

Effects of Different Polystyrene Concentrations on the Bioaccumulation of As, Cd, and Pb in Ruditapes Philippinarum | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effects of Different Polystyrene Concentrations on the Bioaccumulation of As, Cd, and Pb in Ruditapes Philippinarum Zhen Gao, Zhe Sun, Deyi Wang, yuyu jia This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1237924/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Microplastics constitute a new type of environmental pollutant with a particle size of less than 5 mm that can combine with heavy metals to enter organisms such as clams, and the impact of the combination of microplastics and heavy metals on the marine organism Ruditapes philippinarum remains unclear. Therefore, this problem is analyzed the effects of different concentrations (0, 0.1, 1, 5, and 10% w/w ) of polystyrene (PS) and three heavy metals (arsenic (As), cadmium (Cd), and lead (Pb)) (20 and 200 µg/L) on the bioaccumulation of the heavy metals in clams, and the effects of time for PS (0, 1, and 5%)-bound As, Cd, and Pb (ACP). We found that different concentrations of PS exerted different effects on the absorption of heavy metals by clams. A low concentration of PS (0.1%, 1%) inhibited heavy-metal bioaccumulation, while a high concentration of PS (5%, 10%) promoted their accumulation. Whether or not PS was added, the concentration of As, Cd, and Pb in clams increased with time. The relative amount bioaccumulated in clams was As > Cd > Pb, and we demonstrated a correlation between the concentration of heavy metals in clams and the concentration of PS in seawater. When exposed to a low concentration of PS, the concentrations of As and Cd in clams diminished with the concentration of PS in sea water; and when exposed to high concentrations of PS, the concentrations of Cd and Pb in clams appeared also to be reduced with increasing PS concentrations. Clam Arsenic Cadmium Lead Microplastics Bioaccumulation Figures Figure 1 Figure 2 Figure 3 1. Introduction Microplastics (MPs) are very small plastic particles or fragments that are difficult to visualize macroscopically (Kurniawan, et al. 2021 ), with a particle size of generally less than 5 mm (Cai et al. 2020 ). Importantly, MPs pose a potential threat to the environment (Kurniawan, et al. 2021 ), and they can enter the marine ecosystem directly or indirectly (Kurniawan, et al. 2021 ). Synthetic fibers, tires, and city dust are the principal sources of MPs in the ocean (Liu et al. 2021 ).Neto et al. ( 2020 ) tested 965 fish samples from the southeastern and southern coasts of Brazil, and we found that all contained plastics in their bodies. MPs have been shown to reduce the energy storage capacity of the marine organism Mytilus galloprovincialis by altering the activity of digestive enzymes (Trestrail et al. 2021 ), and another study revealed that most seabirds on the Korean Peninsula ingested plastic debris (Nam et al. 2021 ). Thus, controlling marine MPs pollution has become a timely challenge (Haward 2018 ). Polystyrene (PS) is often used as disposable cups, styrofoam, and disposable containers, which are difficult to recover after being discarded and easy to deposited on seashore (Kurniawan, et al. 2021 ), it poses a threat to the survival and safety of marine organisms, so PS is selected as the research object of this experiment. Arsenic (As) is ubiquitous in the environment, both naturally circulated and due to human input; it can also bioaccumulate and biotransform in organisms (Hirano 2020 ). Cadmium (Cd) exists in the environment as a pollutant (Genchi et al. 2020 ) that exerts a negative impact on plant growth (Haider et al. 2021 ), and Cd exposure has been shown to lead (Pb)to cell death in the zebrafish brain (Monaco et al. 2017 ). Pb is a globally distributed heavy-metal pollutant (Sary et al. 2012), that has been known for millennia (Cheung et al. 2017); and exposure can Pb to disorders of various bodily systems, induce inflammation, and cause chronic and acute poisoning (Boskabady et al. 2018 ). The concentrations of copper (Cu), Pb, zinc (Zn), and Cd have shown an upward trend in recent decades, and these heavy metals are found, for example, in the surface sediments of the Bohai Sea of China (Duan et al. 2017); and As, Cd, and Pb particularly showed slight-to-moderate pollution of this body of water. However, Cd and As can engender potentially considerable ecological risks due to their high toxicity (X. Wang et al. 2020 ). Yang et al. found that As hazard quotients in marine fish were high, and this requires our determined attention (Yang et al. 2021 ). Investigators have in fact uncovered As, Cd, and Pb in 30 types of marine fish (Kumar et al. 2021 ); and their muscle content of Pb was higher than in the liver (Sary et al. 2012). Chiocchetti et al. have stated that to provide healthy seafood to consumers, we need to pay greater attention to the toxic heavy metals and metalloids, such as mercury (Hg), Cd, As, and Pb in seafood (Chiocchetti et al. 2017 ). There are manifold environmental pollution sources, so it is meaningful to select the common metals As, Cd and Pb as the experimental heavy metal exposure to explore the impact on the survival of clams. When heavy metals in seawater accumulate to a significant extent, they also adsorb MPs (Gao et al. 2019 ). In some cases, the concentrations of heavy metals in MPs collected from beaches and sediments were higher than those in local estuarine sediments (Holmes et al. 2012 ). The average concentrations of Pb and Cu in MPs are also higher in rivers than in surface water because MPs have the ability to absorb heavy metals from the aquatic environment (Purwiyanto et al. 2020 ). Also, the metal concentrations in PS and polyvinyl chloride are 800 times higher than in the surrounding environment (Naqash et al. 2020 ). What effects the co-exposure of MPs and heavy metals have on organisms is far from clear, and thus many scholars have analyzed the combined effects of MPs and heavy metals on organisms. The total As concentration in earthworms exposed to As(V)+MPs was significantly lower than that for As(V)-alone (Wang et al. 2019 ). Therefore, co-exposure of MPs and heavy metals can precipitate synergistic, antagonistic, or intensifying effects on organisms (Cao et al. 2021 ). However, there are relatively few studies on the co-exposure to MPs and heavy metals in marine organisms. Meanwhile clams are seafood that human beings like, and their safety is very important. Therefore, the combined exposure of PS and heavy metals was selected to observe its effect on the enrichment of heavy metals in marine organism clams. In this study, we: (1) investigated the effects of co-exposure to As, Cd, Pb, and a series of concentrations of PS on the bioaccumulation of heavy metals in Ruditapes philippinarum ; (2) explored the effects of adding PS to the absorption of heavy metals by Ruditapes philippinarum over time; and (3) probed the potential relationship between the concentration of PS in seawater and the concentration of PS in Ruditapes philippinarum . 2. Materials And Methods 2.1 Clam collection and reagents Ruditapes philippinarum clams, 2.0–3.0 cm in size, were purchased from a coastal aquaculture farm in Yantai City, Shandong Province, China. The clams were taken to the laboratory were randomly and evenly placed in sixteen the 3.5-litre stainless steel basins of 30 specimens per tank, with about 1 L of naturally filtered seawater at 20°C. We removed seawater from Yantai Red Lip Beach and changed it once per day, oxygenated the seawater with an oxygen pump (SEBO) on a 24-h cycle, and maintained the clams under a 12-h light:12-h dark cycle. The clams were acclimated to the natural seawater for 14 days under laboratory conditions prior to being placed in the experiment. PS was the common MPs in marine environment (Xiaopeng Zhu et al. 2020 ). PS has a stronger pollutant adsorption capacity than other MPs at room temperature (Alimi et al. 2018 ). The experimental microplastic was 100 nm, that was purchased from Dongguan Zhangmutou Huachuang plastic raw material commercial firm, Guangdong, China. As (Arsenic +5 1000 µg/mL in H 2 O) was purchased from o2Si (Charleston, USA). Both Cd (CdCl 2 ·2.5H 2 O, CAS 7790-78-5) and Pb (Pb (NO 3 ) 2 , CAS 10099-74-8) for analytical pure, were purchased from the Shanghai test (SCR, Shanghai, China). 2.2 Experimental conditions for co-exposure The fresh seawater and a small amount of sand were taken from honglip beach in Yantai, China, the background concentration of As, Cd and Pb were 7.998, 0.086, and 0.032 µg/L, respectively. Fresh seawater with a little sand was taken to change the water for clams once a day. Approximately 1L seawater containing sand was weighed into each stainless-steel basin (3.5L). PS particles were evenly mixed with seawater, then add a certain concentration of As, Cd, and Pb. There were three experimental variables: (1) two As, Cd, and Pb concentrations (20 and 200 µg/L(ppb)), combined with previous studies, the concentration range of As, Cd, and Pb is 0.01-248.8 µg/L in the surface seawater of the Bohai Sea (Tian et al. 2020 ), therefore, the experiment uniformly selects the concentration of heavy metals is 20 ppb, and on this basis, the concentration is increased by an order of magnitude; (2) five doses of PS (0, 0.1, 1, 5, and 10% [ w/w ]), the concentration of PS was refer to experiment ((Kurniawan, et al. 2021 ), (Liu et al. 2021 ), (Wang et al. 2020 )); and (3) five sampling times (1 d, 3 d, 5 d, 7 d, and 14 d) for the exposure test. Therefore, a total of 10 treatments were applied, with each treatment having four replicates. On the 14th day of the exposure experiment, we collected 10 groups of clams that were co-exposed to 20 ppb or 200 ppb ACP (As, Cd, and Pb) with 0, 0.1, 1, 5, or 10% PS, respectively, which with five parallel experiments. We also conducted clam collection over time (1, 3, 5, 7, and 14 d) for co-exposure to 200 ppb ACP (As, Cd, and Pb) with 0%, 1%, or 5% PS, respectively. After cleaning the clamshells with tap water and deionized water and wiping off the water with paper toweling, we removed all of the clam meat, and the rest of the digestive gland was cut up and homogenized (MY-20 in a handheld tissue grinder (Shanghai Jingxin Industrial Development Co., Ltd.). The samples were then stored in a freezer at −20°C prior to testing. 2.3 Analysis of heavy matal in samples The concentration of As Cd, and Pb in Ruditapes philippinarum were analyzed by ICP-MS (Agilent 7700×). The process of sample digestion was to weigh about 1.5g samples, add 8 mL HNO 3 (70% wt, ≥99.999% metal basis) and 2 mL H 2 O 2 (30% wt, guaranteed reagent grade) in polytetrafluoroethylene digestion tank, heated it to 150 ℃ within 10 minutes, then heated it to 180℃ in 15 minutes, and hold it at 180℃ for 60 minutes in microwave digestion machine (MDS-6G). After digestion, the sample was incubated to 25 ml using ultrapure water and filtered with a 0.22 µm acetate cellulose membrane. The certified reference material (NCRM, GBW10210 Salmon lyophilized powder, Tanmo Quality Inspection Technology Co., Ltd, Jiangsu, China) was used to verify the reliability of the analysis method. The recovery rate of As, Cd, and Pb rate were estimated to 95±5%, 125±4%, and 97±5%, respectively. To ensure quality guarantee of the data, the method blank was included during sample analysis. 2.4 Statistical analysis The data were sorted and statistically analyzed by Microsoft Excel 2019 and SPSS statistics 25.0 software. The experimental data were expressed by the arithmetic mean ± standard deviation (mean ± SD) (n = 5). One-way ANOVA and LSD were used to test the significant difference (P < 0.05). Origin 2019 software was used for correlation analysis and mapping. 3. Results And Discussion 3.1 PS on the absorption of As, Cd, and Pb by clams As shown in Fig. 1 and Table S1, we analyzed the concentrations of As, Cd, and Pb (ACP) in clams after 14 days of co-exposure with 20 ppb or 200 ppb As, Cd, and Pb and with different concentrations of PS (0%, 0.1%, 1%, 5%, and 10%) ( w/w ). We found that the different concentrations of PS could promote or inhibit the absorption of heavy metals (As, Cd, and Pb) by clams. Compared with 0% PS, 0.1%, 1%, 5%, and 10% PS inhibited the bioaccumulation of 200 ppb As by clams; and both 0.1% and 1% PS inhibited clam accumulation at 20 or 200 ppb Cd and Pb (Fig. 1BC). In conclusion, low concentrations of PS (0.1% and 1%) inhibit the bioaccumulation of heavy metals (As, Cd, and Pb) in clams. Similar results were also observed when the co-exposure of As(V) and MPs (2000 mg MP kg −1 dry soil=0.2% [ w/w ]) led to a diminution in As(V) and in the accumulation of total As in the earthworm intestine, with the possible reason being that MPs may reduce the bioavailability of As by adsorbing/binding As(V) (Wang et al. 2019 ). Other similar research showed that the exposure to MPs (PSNPs) (10 mg/L=0.001% [ w/w ]) at low concentrations promoted the growth of wheat, and MPs partially reduced the toxicity of Cd to wheat. Thus, MPs may have been used as a carrier of Cd in wheat, decreasing Cd toxicity as the concentration of Cd in PSNPs-Cd culture medium was lower than that in the Cd solution alone, and metabonomic analysis further showed that the increase in carbohydrate and amino acid metabolism caused by PSNPs partially alleviated the toxicity of Cd to wheat (Lian et al. 2020 ). Although there was no adverse effect on the survival and growth of the fish, co-exposure to MPs (50 or 500 µg L −1 ) and Cd (50 µg L −1 ) for 30 days resulted in reduced Cd bioaccumulation in the discus fish due to the presence of MPs; and co-exposure also precipitated severe oxidative stress and stimulated innate immunity (Wen et al. 2018 ). Combined exposure to MPs and Cd showed antagonistic effect on zebrafish embryos at a low MP concentration in one study, and compared with the absence of MPs, the study showed lower mortality, higher heart rate, and improved body length (Zhang et al. 2020 ). Because there are limited studies on the combination of heavy metals and MP in aquatic organisms, this combination of toxicants needs to be evaluated further. PS at 5% significantly promoted the absorption of 20 and 200 ppb Cd by clams (P < 0.05), and 10% PS also promoted absorption at 200 ppb Cd (Fig. 1 B); 5% PS significantly promoted absorption of 20 ppb Pb in clams (P < 0.05); and 10% PS also promoted it at 200 ppb Pb (Fig. 1 C). A high concentration of PS (5% and 10%) promoted the heavy-metal (Cd and Pb) accumulation in clams, and 5% PS was even more augmentative. Studies have shown that high doses of PS enhance phytotoxicity for co-exposure of maize seeds to 10% HDPE, which is similar to our results (Wang et al. 2020 ). Plastics may also become carriers of some contaminants (Alimi et al. 2018 ), and most collected oysters bear microplastic particles (Zhu et al. 2020 ). Therefore, ingestion of MPs allocates a pathway for the metal transfer to organisms (Naqash et al. 2020 ). However, this is species dependent; as in the experimental exposure to MPs in mussels, it was found that mussels can excrete most microplastic particles within a certain period of time, and that the burden of MPs in large mussels is much less than that in small mussels (Weber et al. 2021 ). In conclusion, we found that a low concentration of MPs inhibited the accumulation by clams of heavy metals, and that high-concentration MPs exacerbated their accumulation. When zebrafish embryos were co-exposed to MPs and Cd, the investigators found that low concentrations of MPs (0.05, 0.1 mg/L) showed antagonistic effects on physiological indices in zebrafish, while high-concentration MPs (1, 5, 10 mg/L) showed synergistic sublethal toxicity (Zhang et al. 2020 ); and this is consistent with the present chapter. However, after clams were exposed to 20 ppb ACP+ PS for 14 days, 0.1% and 1% PS promoted the accumulation of As in clams, but a PS of 5% and 10% inhibited clam accumulation of 20 ppb As (Fig. 1 A). These results are similar to previous studies: when maize seeds were co-exposed to Cd and MPs, it was revealed that a lower concentration of polyethylene particles (0.1%, 1%) ( w/w ) promoted the bioavailability of heavy metals, while a higher concentration (10%) ( w/w ) inhibited the bioavailability of heavy metals (Wang et al. 2020 ). Therefore, the amount and number of heavy metals taken up by clams may exert differential effects on various concentrations of PS or PS combined with different types of heavy metals. 3.2 Effect of adding PS on the absorption of heavy metals by clams over time Clams were co-exposed to 0%, 1%, or 5% PS and 200 ppb As, Cd, and Pb (ACP) The concentrations of As, Cd, and Pb (wet weight) in clams were measured 1, 3, 5, 7, and 14 days after the start of the exposure experiment (Fig. 2 and Table S2), and we ascertained that regardless of whether or not PS was added, the concentrations of As, Cd, and Pb in the clams were elevated with time, and the heavy-metal absorptive capacity of the clams was As > Cd > Pb (Fig. 2 ). The high content of As was related to the elevated background concentration of As in clams, which is consistent with the tendency for increased metal levels in clams with time after a single heavy-metal exposure. Ruditapes philippinarum also showed a time-dependence after exposure to Cd (4 and 40 µg/L) for 70 days. By the middle of the Cd-exposure period, the Cd concentration in clams increased with exposure time, and then gradually decreased during the middle and later stages (Zhao et al. 2014 ). When Ruditapes philippinarum clams were exposed to Pb (10 and 100 µg/L) for 7 days, the Pb concentrations also increased with time commensurately (Aouini et al. 2018 ). MPs absorb heavy metals from the aquatic environment (Purwiyanto et al. 2020 ). Although the content of some heavy metals in seawater is low, they can still adsorb MPs when they gather to a certain extent (Gao et al. 2019 ). Also, the longer the exposure time, the higher the concentration of heavy metals bound by MPs in seawater (Gao et al. 2019 ); and the addition of MPs exhibits a significant effect on the chemical forms of many heavy metals (Yu et al. 2021 ). According to Holmes et al., metal adsorption is conducted through the interaction between divalent cations such as Cu 2+ , Pb 2+ , and Cr 2+ , and oxygen anions such as Cr 2 O 4 2− that possess charged or polar regions on the plastic surface; and via the nonspecific interactions between neutral metal-organic complexes and the hydrophobic surface of bulk plastic medium (Holmes et al. 2012 ). When the interaction between MPs and heavy metals is via physical adsorption, the chemical bond is weaker and the metals can be more easily released into the aquatic ecosystem again (Purwiyanto et al. 2020 ). However, PS adsorb Cu and Cd principally by chemical adsorption (Xu et al. 2021 ). 3.3 Promotion or inhibition of metal accumulation in clams due to PS Clams exposed to 200 ppb ACP and 1% PS exhibited augmented bioaccumulation of As on days 1, 3, and 5, but showed inhibition on days 7 and 14— indicating that ACP gradually inhibited 20-ppb As absorption with time and 5% PS inhibited As accumulation within 14 days (Table 1 ). Adding 1% or 5% of PS generally inhibited Cd accumulation in clams over time, and only 5% PS promoted Cd accumulation in clams on the 14th day (Table 1 ). The addition of 1% PS promoted Pb uptake on the first day, but inhibited it from 3 to 14 days, and the addition of 5% PS stimulated the uptake of Pb (Table 1 ). The bioaccumulation of heavy metals in clams due to MPs should be observed over a range of times because a single timepoint produces insufficient and potentially inaccurate data. Overall, 1% PS inhibited the accumulation of Cd and Pb within 14 days and gradually inhibited the uptake of As; while 5% PS inhibited the absorption of As and Cd and promoted the bioaccumulation of Pb within 14 days (Fig. 3 ). 3.4 Correlation analysis Using correlation analysis, we found that there was also a correlation between the concentration of heavy metals in clams and the concentration of MPs in seawater. At low concentrations of PS plus 20 ppb ACP, As concentration was positively correlated with PS concentration in clams (r 2 = 0.335, P < 0.05) (Fig. 3 A); however, Cd and Pb were not correlated with PS concentration (Fig. 3CE). Simultaneous exposure to low-concentration PS and 200 ppb ACP resulted in a negative correlation between the concentration of PS and concentrations of As and Cd in clams (Fig. 3 AC), although there was no correlation with the concentration of Pb (Fig. 3 E). These data thus illustrated an inverse relationship between As/Cd and PS—i.e., when clams were exposed to 200 ppb As or Cd and a concentration of PS below 1% ( w/w ), we observed that a higher concentration of PS was concomitant with a lower concentration of As or Cd. When exposed to high-concentration PS and 20 ppb ACP, the concentrations of Cd and Pb in clams were significantly negatively correlated with the concentration of PS in seawater(r D 2 =0.725, p<0.05) (r F 2 =0.647, p<0.05) (Fig. 3DF); i.e., when clams were co-exposed to high concentration PS (5–10%) and 20 ppb Cd and Pb, the concentration of Cd and Pb diminished with increasing PS concentration. Table 2 depicts the correlation between a series of PS concentrations and the bioaccumulation of heavy-metal concentrations in clams. We observed a negative correlation between the concentration of PS and 20 or 200 ppb As (r = −0.237, p > 0.05; r = −0.718, p 0.05; r =0.341, p > 0.05). Also, the concentration of PS was negatively correlated with the 20 ppb Pb (r = -0.331, p > 0.05) and 200 ppb Pb (r = 0.038, p > 0.05) in clams. PS can affect the metabolic responses of mussels, but the effect is small. MPs can bioaccumulate in shellfish (oysters)—usually in the form of fibers in gills and mantle—and they can be involved in the adsorption and precipitation of trace metals in oysters. Researchers also found that the in vivo concentration of MPs was correlated with the in vivo concentrations of Cd, chromium (Cr), Cu, and Pb, indicating that there exists a potential in-vivo interaction between MPs and trace metals (Zhu et al. 2020 ). However, the correlations between the concentration of MPs in water and the concentrations of MPs in organisms, and between the concentrations of heavy metals bound to MPs in organisms and the concentration of heavy metals in the organisms have not been fully elucidated and thus require further investigation. 4. Conclusion PS affected the accumulation of As, Cd, and Pb in Ruditapes philippinarum . Different concentrations of PS exerted different effects on the absorption of heavy metals by clams. A low concentration of PS (0.1%, 1%) inhibited heavy-metals (As, Cd, and Pb) bioaccumulation, while a high concentration of PS (5%, 10%) promoted their accumulation. Whether or not PS was added, the concentration of As, Cd, and Pb in clams increased with time. The relative amount bioaccumulated in clams was As > Cd > Pb. The bioaccumulation of heavy metals in clams due to MPs should be observed over a range of times because a single timepoint produces insufficient and potentially inaccurate data. 1% PS inhibited the accumulation of Cd and Pb within 14 days and gradually inhibited the uptake of As; while 5% PS inhibited the absorption of As and Cd and promoted the bioaccumulation of Pb within 14 days. When exposed to a low concentration of PS, the concentrations of As and Cd in clams diminished with the concentration of PS in sea water; and when exposed to high concentrations of PS, the concentrations of Cd and Pb in clams appeared also to be reduced with increasing PS concentrations. Declarations Acknowledgement We would like to thank Prof. Yuyu Jia and Deyi Wang for his constructive comments. Author contribution The author Zhen Gao and Zhe Sun were mainly responsible for experimental research, and authors of Yuyu Jia and Deyi Wang were mainly responsible for coaching. Funding This work was financially supported by the National Natural Science Foundation of China (Grant No. 31900938), the Natural Science Foundation of Shandong Province (Grant No. ZR2019BB073). Data Availability: Not applicable Ethics approval and consent to participate : Not applicable Consent for publication : Not applicable Competing interests : The authors declare no conflict of interest. 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Bull Environ Contam Toxicol 89:82–85. https://doi.org/10.1007/s00128-012-0646-3 Tian K, Wu QM, Liu P, Hu WY, Huang B, Shi B, Zhou YQ, Kwon BO, Choi K, Ryu J, Khim JS, Wang TY (2020) Ecological risk assessment of heavy metals in sediments and water from the coastal areas of the Bohai Sea and the Yellow Sea. Environ Int 136:105512. https://doi.org/10.1016/j.envint.2020.105512 Trestrail C, Walpitagama M, Miranda A, Nugegoda D, Shimeta J (2021) Microplastics alter digestive enzyme activities in the marine bivalve, Mytilus galloprovincialis . Sci Total Environ 779:146418. https://doi.org/10.1016/j.scitotenv.2021.146418 Wang FY, Zhang XQ, Zhang SQ, Zhang SW, Sun YH (2020) Interactions of microplastics and cadmium on plant growth and arbuscular mycorrhizal fungal communities in an agricultural soil. Chemosphere 254:126791. https://doi.org/10.1016/j.chemosphere.2020.126791 Wang FY, Zhang XQ, Zhang SQ, Zhang SW, Adams CA, Sun YH (2020) Effects of Co-Contamination of Microplastics and Cd on Plant Growth and Cd Accumulation. Toxics 8(36):1–12. https://doi.org/10.3390/toxics8020036 Wang HT, Ding J, Xiong C, Zhu D, Li G, Jia XY, Zhu YG, Xue XM (2019) Exposure to microplastics lowers arsenic accumulation and alters gut bacterial communities of earthworm Metaphire californica. Environ Pollut 251:110–116. https://doi.org/10.1016/j.envpol.2019.04.054 Wang XJ, Fu RL, Li HL, Zhang Y, Lu MQ, Xiao K, Zhang XL, Zheng CM, Xiong Y (2020) Heavy metal contamination in surface sediments: A comprehensive, large-scale evaluation for the Bohai Sea, China. Environ Pollut 260:113986. https://doi.org/10.1016/j.envpol.2020.113986 Weber A, Jeckel N, Weil C, Umbach S, Brennholt N, Reifferscheid G (2021) Wagner M, Ingestion and toxicity of polystyrene microplastics in freshwater bivalves. Environ Toxicol Chem 40(8):2247–2260. https://doi.org/10.1002/etc.5076 Wei Q, Hu CY, Zhang RR, Gu YY, Sun AL, Zhang ZM, Shi XZ, Chen J, Wang TZ (2021) Comparative evaluation of high-density polyethylene and polystyrene microplastics pollutants: Uptake, elimination and effects in mussel. Mar Environ Res 169:105329. https://doi.org/10.1016/j.marenvres.2021.105329 Wen B, Jin SR, Chen ZZ, Gao JZ, Liu YN, Liu JH, Feng XS (2018) Single and combined effects of microplastics and cadmium on the cadmium accumulation, antioxidant defence and innate immunity of the discus fish ( Symphysodon aequifasciatus ). Environ Pollut 243:462–471. https://doi.org/10.1016/j.envpol.2018.09.029 Xu GH, Liu Y, Yu Y (2021) Effects of polystyrene microplastics on uptake and toxicity of phenanthrene in soybean. Sci Total Environ 783:147016. https://doi.org/10.1016/j.scitotenv.2021.147016 Yang LP, Guo MC, Xin CL, Ren XF, Wang L, Liu YJ (2021) Comparison of trace element concentrations in freshwater fish and marine fish consumed in Shandong Province, China, and their potential risks to human health. Mar Pollut Bull 165:112114. https://doi.org/10.1016/j.marpolbul.2021.112114 Yu H, Zhang Z, Zhang Y, Fan P, Xi BD, Tan WB (2021) Metal type and aggregate microenvironment govern the response sequence of speciation transformation of different heavy metals to microplastics in soil. Sci Total Environ 752:141956. https://doi.org/10.1016/j.scitotenv.2020.141956 Zhang R, Wang M, Chen XP, Yang CM, Wu LL (2020) Combined toxicity of microplastics and cadmium on the zebrafish embryos ( Danio rerio ). Sci Total Environ 743:140638. https://doi.org/10.1016/j.scitotenv.2020.140638 Zhao LQ, Zhang Y, Liang J, Xu X, Wang H, Yang F, Yan XW (2014) Environmental Cadmium exposure impacts physiological responses in Manila clams. Biol Trace Elem Res 159:241–253. https://doi.org/10.1007/s12011-014-9975-x Zhu XT, Qiang LY, Shi HH, Cheng JP (2020) Bioaccumulation of microplastics and its in vivo interactions with trace metals in edible oysters. Mar Pollut Bull 154:111079. https://doi.org/10.1016/j.marpolbul.2020.111079 Zhu XP, Ran W, Teng J, Zhang C, Zhang WJ, Hou CW, Zhao JM, Qi XT, Wang Q (2020) Microplastic Pollution in Nearshore Sediment from the Bohai Sea Coastline. Bull Environ Contam Toxicol 107:665–670. https://doi.org/10.1007/s00128-020-02866-1 Tables Table.1 The mean concentration of heavy metals in Ruditapes philippinarums after co-exposure of different concentrations of PS As Cd Pb 1% PS 5% PS 1% PS 5% PS 1% PS 5% PS 1 d 1.12±0.01 0.74±0.01 0.85±0.17 0.95±0.12 1.33±0.38 1.21±0.05 3 d 1.10±0.14 0.92±0.14 0.57±0.08 0.63±0.07 0.71±0.13 1.10±1.11 5d 1.12±0.07 0.68±0.45 0.77±0.16 0.78±0.08 0.87±0.09 0.99±0.40 7d 0.93±0.26 0.88±0.07 0.59±0.13 0.80±0.18 0.84±0.10 1.74±0.18 14d 0.50±0.43 0.76±0.03 0.71±0.02 1.25±0.12 0.77±0.10 1.06±0.33 Table.2 Pearson correlation between PS and heavy metal in Ruditapes philippinarums As Cd Pb 20ppb 200ppb 20ppb 200ppb 20ppb 200ppb PS -0.237 -0.718 0.102 0.341 -0.331 0.038 Supplementary Files Graphical.jpg highlight.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-1237924","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":83157344,"identity":"269f3f6c-28d3-4f67-bc11-be91267f636f","order_by":0,"name":"Zhen Gao","email":"","orcid":"","institution":"Yantai University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zhen","middleName":"","lastName":"Gao","suffix":""},{"id":83157345,"identity":"80e33060-08fc-4601-9718-df41a0bb747d","order_by":1,"name":"Zhe Sun","email":"","orcid":"","institution":"Yantai University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zhe","middleName":"","lastName":"Sun","suffix":""},{"id":83157346,"identity":"1f3a6147-c9f3-46fd-9733-7b198e4e523b","order_by":2,"name":"Deyi Wang","email":"","orcid":"","institution":"Yantai University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Deyi","middleName":"","lastName":"Wang","suffix":""},{"id":83157347,"identity":"06f68a37-b757-4ed7-8344-e1754accb43a","order_by":3,"name":"yuyu jia","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5ElEQVRIiWNgGAWjYDADPgbGxgMMFRJy8kRrYWNgbDjAcMbC2LCBeC0MDAcY2yoSGQ4QUGkukfzscWGbXR4b++GGAx/nSSQwNjA/fHQDjxbLGWnmxjPbkovZeBIbDs7cJpHHzsBmbJyDR4vBjQQzad425sQ2hsSGw7zbJIoZG3jYpPFrSf8G1FKf2Mb/sOHw3zkSiQ0HCGrJAdlyOLENqPgwYwMxWs68KZPmOXccqOVhw8GeYxLGhs2E/HI8fZs0T1l1Yj9/+sMHP2rq5OTZmx8+xqcFC2AmTfkoGAWjYBSMAiwAAKxHS5IU7hcoAAAAAElFTkSuQmCC","orcid":"","institution":"Yantai University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"yuyu","middleName":"","lastName":"jia","suffix":""}],"badges":[],"createdAt":"2022-01-07 09:23:34","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1237924/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1237924/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":18258591,"identity":"fbc2238c-e598-47f6-900d-1787afd76026","added_by":"auto","created_at":"2022-02-15 23:18:05","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":130156,"visible":true,"origin":"","legend":"\u003cp\u003eThe mean concentration of As, Cd and Pb with different concentrations of PS in \u003cem\u003eRuditapes philippinarums\u003c/em\u003e\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-1237924/v1/214eff844db4dd2f31c92d42.jpeg"},{"id":18258618,"identity":"93aedfc0-6e6a-488e-af6c-7d154e1cb012","added_by":"auto","created_at":"2022-02-15 23:21:05","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":96055,"visible":true,"origin":"","legend":"\u003cp\u003eConcentration of As, Cd and Pb in different concentrations of PS over time\u003c/p\u003e\u003cp\u003e\u0026nbsp;(A) 0% PS + 200ppb ACP; (B) 1% PS + 200ppb ACP; (C) 10% PS + 200ppb ACP\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-1237924/v1/fb1f29bf4f01a347133a9174.jpeg"},{"id":18258593,"identity":"db3a3511-6b82-40a1-aeea-d2885c044562","added_by":"auto","created_at":"2022-02-15 23:18:05","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":248638,"visible":true,"origin":"","legend":"\u003cp\u003eCorrelation between the concentration of As, Cd and Pb and PS\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-1237924/v1/49264bb09448e05912354949.jpeg"},{"id":21644151,"identity":"1b1fe314-985a-4306-a74d-7de76034d9fd","added_by":"auto","created_at":"2022-05-19 04:40:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":570262,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1237924/v1/72cc2401-cf50-4de4-a39b-d0d743dc8ede.pdf"},{"id":18258595,"identity":"25821ea1-a119-44da-93ad-881ff25946e1","added_by":"auto","created_at":"2022-02-15 23:18:06","extension":"jpg","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":2661681,"visible":true,"origin":"","legend":"","description":"","filename":"Graphical.jpg","url":"https://assets-eu.researchsquare.com/files/rs-1237924/v1/2cec3e0ca23977467ada6d76.jpg"},{"id":18258592,"identity":"bcb1f279-99de-41b0-b886-0936fb2012eb","added_by":"auto","created_at":"2022-02-15 23:18:05","extension":"docx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":14689,"visible":true,"origin":"","legend":"","description":"","filename":"highlight.docx","url":"https://assets-eu.researchsquare.com/files/rs-1237924/v1/799cdb4d0038c9573ae2dc6b.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003eEffects of Different Polystyrene Concentrations on the Bioaccumulation of As, Cd, and Pb in Ruditapes Philippinarum\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eMicroplastics (MPs) are very small plastic particles or fragments that are difficult to visualize macroscopically (Kurniawan, et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), with a particle size of generally less than 5 mm (Cai et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Importantly, MPs pose a potential threat to the environment (Kurniawan, et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), and they can enter the marine ecosystem directly or indirectly (Kurniawan, et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Synthetic fibers, tires, and city dust are the principal sources of MPs in the ocean (Liu et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).Neto et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) tested 965 fish samples from the southeastern and southern coasts of Brazil, and we found that all contained plastics in their bodies. MPs have been shown to reduce the energy storage capacity of the marine organism \u003cem\u003eMytilus galloprovincialis\u003c/em\u003e by altering the activity of digestive enzymes (Trestrail et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), and another study revealed that most seabirds on the Korean Peninsula ingested plastic debris (Nam et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Thus, controlling marine MPs pollution has become a timely challenge (Haward \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Polystyrene (PS) is often used as disposable cups, styrofoam, and disposable containers, which are difficult to recover after being discarded and easy to deposited on seashore (Kurniawan, et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), it poses a threat to the survival and safety of marine organisms, so PS is selected as the research object of this experiment.\u003c/p\u003e \u003cp\u003eArsenic (As) is ubiquitous in the environment, both naturally circulated and due to human input; it can also bioaccumulate and biotransform in organisms (Hirano \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Cadmium (Cd) exists in the environment as a pollutant (Genchi et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) that exerts a negative impact on plant growth (Haider et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), and Cd exposure has been shown to lead (Pb)to cell death in the zebrafish brain (Monaco et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Pb is a globally distributed heavy-metal pollutant (Sary et al. 2012), that has been known for millennia (Cheung et al. 2017); and exposure can Pb to disorders of various bodily systems, induce inflammation, and cause chronic and acute poisoning (Boskabady et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The concentrations of copper (Cu), Pb, zinc (Zn), and Cd have shown an upward trend in recent decades, and these heavy metals are found, for example, in the surface sediments of the Bohai Sea of China (Duan et al. 2017); and As, Cd, and Pb particularly showed slight-to-moderate pollution of this body of water. However, Cd and As can engender potentially considerable ecological risks due to their high toxicity (X. Wang et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Yang et al. found that As hazard quotients in marine fish were high, and this requires our determined attention (Yang et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Investigators have in fact uncovered As, Cd, and Pb in 30 types of marine fish (Kumar et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2021\u003c/span\u003e); and their muscle content of Pb was higher than in the liver (Sary et al. 2012). Chiocchetti et al. have stated that to provide healthy seafood to consumers, we need to pay greater attention to the toxic heavy metals and metalloids, such as mercury (Hg), Cd, As, and Pb in seafood (Chiocchetti et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). There are manifold environmental pollution sources, so it is meaningful to select the common metals As, Cd and Pb as the experimental heavy metal exposure to explore the impact on the survival of clams.\u003c/p\u003e \u003cp\u003eWhen heavy metals in seawater accumulate to a significant extent, they also adsorb MPs (Gao et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In some cases, the concentrations of heavy metals in MPs collected from beaches and sediments were higher than those in local estuarine sediments (Holmes et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The average concentrations of Pb and Cu in MPs are also higher in rivers than in surface water because MPs have the ability to absorb heavy metals from the aquatic environment (Purwiyanto et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Also, the metal concentrations in PS and polyvinyl chloride are 800 times higher than in the surrounding environment (Naqash et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). What effects the co-exposure of MPs and heavy metals have on organisms is far from clear, and thus many scholars have analyzed the combined effects of MPs and heavy metals on organisms. The total As concentration in earthworms exposed to As(V)+MPs was significantly lower than that for As(V)-alone (Wang et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Therefore, co-exposure of MPs and heavy metals can precipitate synergistic, antagonistic, or intensifying effects on organisms (Cao et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). However, there are relatively few studies on the co-exposure to MPs and heavy metals in marine organisms. Meanwhile clams are seafood that human beings like, and their safety is very important. Therefore, the combined exposure of PS and heavy metals was selected to observe its effect on the enrichment of heavy metals in marine organism clams.\u003c/p\u003e \u003cp\u003eIn this study, we: (1) investigated the effects of co-exposure to As, Cd, Pb, and a series of concentrations of PS on the bioaccumulation of heavy metals in \u003cem\u003eRuditapes philippinarum\u003c/em\u003e; (2) explored the effects of adding PS to the absorption of heavy metals by \u003cem\u003eRuditapes philippinarum\u003c/em\u003e over time; and (3) probed the potential relationship between the concentration of PS in seawater and the concentration of PS in \u003cem\u003eRuditapes philippinarum\u003c/em\u003e.\u003c/p\u003e"},{"header":"2. Materials And Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Clam collection and reagents\u003c/h2\u003e \u003cp\u003e \u003cem\u003eRuditapes philippinarum\u003c/em\u003e clams, 2.0\u0026ndash;3.0 cm in size, were purchased from a coastal aquaculture farm in Yantai City, Shandong Province, China. The clams were taken to the laboratory were randomly and evenly placed in sixteen the 3.5-litre stainless steel basins of 30 specimens per tank, with about 1 L of naturally filtered seawater at 20\u0026deg;C. We removed seawater from Yantai Red Lip Beach and changed it once per day, oxygenated the seawater with an oxygen pump (SEBO) on a 24-h cycle, and maintained the clams under a 12-h light:12-h dark cycle. The clams were acclimated to the natural seawater for 14 days under laboratory conditions prior to being placed in the experiment.\u003c/p\u003e \u003cp\u003ePS was the common MPs in marine environment (Xiaopeng Zhu et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). PS has a stronger pollutant adsorption capacity than other MPs at room temperature (Alimi et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The experimental microplastic was 100 nm, that was purchased from Dongguan Zhangmutou Huachuang plastic raw material commercial firm, Guangdong, China. As (Arsenic\u003csup\u003e+5\u003c/sup\u003e 1000 \u0026micro;g/mL in H\u003csub\u003e2\u003c/sub\u003eO) was purchased from o2Si (Charleston, USA). Both Cd (CdCl\u003csub\u003e2\u003c/sub\u003e\u0026middot;2.5H\u003csub\u003e2\u003c/sub\u003eO, CAS 7790-78-5) and Pb (Pb (NO\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003e, CAS 10099-74-8) for analytical pure, were purchased from the Shanghai test (SCR, Shanghai, China).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Experimental conditions for co-exposure\u003c/h2\u003e \u003cp\u003eThe fresh seawater and a small amount of sand were taken from honglip beach in Yantai, China, the background concentration of As, Cd and Pb were 7.998, 0.086, and 0.032 \u0026micro;g/L, respectively. Fresh seawater with a little sand was taken to change the water for clams once a day. Approximately 1L seawater containing sand was weighed into each stainless-steel basin (3.5L). PS particles were evenly mixed with seawater, then add a certain concentration of As, Cd, and Pb.\u003c/p\u003e \u003cp\u003eThere were three experimental variables: (1) two As, Cd, and Pb concentrations (20 and 200 \u0026micro;g/L(ppb)), combined with previous studies, the concentration range of As, Cd, and Pb is 0.01-248.8 \u0026micro;g/L in the surface seawater of the Bohai Sea (Tian et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), therefore, the experiment uniformly selects the concentration of heavy metals is 20 ppb, and on this basis, the concentration is increased by an order of magnitude; (2) five doses of PS (0, 0.1, 1, 5, and 10% [\u003cem\u003ew/w\u003c/em\u003e]), the concentration of PS was refer to experiment ((Kurniawan, et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), (Liu et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), (Wang et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e)); and (3) five sampling times (1 d, 3 d, 5 d, 7 d, and 14 d) for the exposure test. Therefore, a total of 10 treatments were applied, with each treatment having four replicates. On the 14th day of the exposure experiment, we collected 10 groups of clams that were co-exposed to 20 ppb or 200 ppb ACP (As, Cd, and Pb) with 0, 0.1, 1, 5, or 10% PS, respectively, which with five parallel experiments. We also conducted clam collection over time (1, 3, 5, 7, and 14 d) for co-exposure to 200 ppb ACP (As, Cd, and Pb) with 0%, 1%, or 5% PS, respectively. After cleaning the clamshells with tap water and deionized water and wiping off the water with paper toweling, we removed all of the clam meat, and the rest of the digestive gland was cut up and homogenized (MY-20 in a handheld tissue grinder (Shanghai Jingxin Industrial Development Co., Ltd.). The samples were then stored in a freezer at \u0026minus;20\u0026deg;C prior to testing.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Analysis of heavy matal in samples\u003c/h2\u003e \u003cp\u003eThe concentration of As Cd, and Pb in \u003cem\u003eRuditapes philippinarum\u003c/em\u003e were analyzed by ICP-MS (Agilent 7700\u0026times;). The process of sample digestion was to weigh about 1.5g samples, add 8 mL HNO\u003csub\u003e3\u003c/sub\u003e (70% wt, \u0026ge;99.999% metal basis) and 2 mL H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e (30% wt, guaranteed reagent grade) in polytetrafluoroethylene digestion tank, heated it to 150 ℃ within 10 minutes, then heated it to 180℃ in 15 minutes, and hold it at 180℃ for 60 minutes in microwave digestion machine (MDS-6G). After digestion, the sample was incubated to 25 ml using ultrapure water and filtered with a 0.22 \u0026micro;m acetate cellulose membrane. The certified reference material (NCRM, GBW10210 Salmon lyophilized powder, Tanmo Quality Inspection Technology Co., Ltd, Jiangsu, China) was used to verify the reliability of the analysis method. The recovery rate of As, Cd, and Pb rate were estimated to 95\u0026plusmn;5%, 125\u0026plusmn;4%, and 97\u0026plusmn;5%, respectively. To ensure quality guarantee of the data, the method blank was included during sample analysis.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Statistical analysis\u003c/h2\u003e \u003cp\u003eThe data were sorted and statistically analyzed by Microsoft Excel 2019 and SPSS statistics 25.0 software. The experimental data were expressed by the arithmetic mean \u0026plusmn; standard deviation (mean \u0026plusmn; SD) (n = 5). One-way ANOVA and LSD were used to test the significant difference (P \u0026lt; 0.05). Origin 2019 software was used for correlation analysis and mapping.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results And Discussion","content":"\u003cdiv class=\"Section2\" id=\"Sec8\"\u003e\n \u003ch2\u003e3.1 PS on the absorption of As, Cd, and Pb by clams\u003c/h2\u003e\n \u003cp\u003eAs shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e and Table S1, we analyzed the concentrations of As, Cd, and Pb (ACP) in clams after 14 days of co-exposure with 20 ppb or 200 ppb As, Cd, and Pb and with different concentrations of PS (0%, 0.1%, 1%, 5%, and 10%) (\u003cem\u003ew/w\u003c/em\u003e). We found that the different concentrations of PS could promote or inhibit the absorption of heavy metals (As, Cd, and Pb) by clams.\u003c/p\u003e\n \u003cp\u003eCompared with 0% PS, 0.1%, 1%, 5%, and 10% PS inhibited the bioaccumulation of 200 ppb As by clams; and both 0.1% and 1% PS inhibited clam accumulation at 20 or 200 ppb Cd and Pb (Fig.\u0026nbsp;1BC). In conclusion, low concentrations of PS (0.1% and 1%) inhibit the bioaccumulation of heavy metals (As, Cd, and Pb) in clams.\u003c/p\u003e\n \u003cp\u003eSimilar results were also observed when the co-exposure of As(V) and MPs (2000 mg MP kg\u003csup\u003e\u0026minus;1\u003c/sup\u003e dry soil=0.2% [\u003cem\u003ew/w\u003c/em\u003e]) led to a diminution in As(V) and in the accumulation of total As in the earthworm intestine, with the possible reason being that MPs may reduce the bioavailability of As by adsorbing/binding As(V) (Wang et al. \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). Other similar research showed that the exposure to MPs (PSNPs) (10 mg/L=0.001% [\u003cem\u003ew/w\u003c/em\u003e]) at low concentrations promoted the growth of wheat, and MPs partially reduced the toxicity of Cd to wheat. Thus, MPs may have been used as a carrier of Cd in wheat, decreasing Cd toxicity as the concentration of Cd in PSNPs-Cd culture medium was lower than that in the Cd solution alone, and metabonomic analysis further showed that the increase in carbohydrate and amino acid metabolism caused by PSNPs partially alleviated the toxicity of Cd to wheat (Lian et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). Although there was no adverse effect on the survival and growth of the fish, co-exposure to MPs (50 or 500 \u0026micro;g L\u003csup\u003e\u0026minus;1\u003c/sup\u003e) and Cd (50 \u0026micro;g L\u003csup\u003e\u0026minus;1\u003c/sup\u003e) for 30 days resulted in reduced Cd bioaccumulation in the discus fish due to the presence of MPs; and co-exposure also precipitated severe oxidative stress and stimulated innate immunity (Wen et al. \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e). Combined exposure to MPs and Cd showed antagonistic effect on zebrafish embryos at a low MP concentration in one study, and compared with the absence of MPs, the study showed lower mortality, higher heart rate, and improved body length (Zhang et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). Because there are limited studies on the combination of heavy metals and MP in aquatic organisms, this combination of toxicants needs to be evaluated further.\u003c/p\u003e\n \u003cp\u003ePS at 5% significantly promoted the absorption of 20 and 200 ppb Cd by clams (P \u0026lt; 0.05), and 10% PS also promoted absorption at 200 ppb Cd (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eB); 5% PS significantly promoted absorption of 20 ppb Pb in clams (P \u0026lt; 0.05); and 10% PS also promoted it at 200 ppb Pb (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eC). A high concentration of PS (5% and 10%) promoted the heavy-metal (Cd and Pb) accumulation in clams, and 5% PS was even more augmentative. Studies have shown that high doses of PS enhance phytotoxicity for co-exposure of maize seeds to 10% HDPE, which is similar to our results (Wang et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). Plastics may also become carriers of some contaminants (Alimi et al. \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e), and most collected oysters bear microplastic particles (Zhu et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). Therefore, ingestion of MPs allocates a pathway for the metal transfer to organisms (Naqash et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). However, this is species dependent; as in the experimental exposure to MPs in mussels, it was found that mussels can excrete most microplastic particles within a certain period of time, and that the burden of MPs in large mussels is much less than that in small mussels (Weber et al. \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eIn conclusion, we found that a low concentration of MPs inhibited the accumulation by clams of heavy metals, and that high-concentration MPs exacerbated their accumulation. When zebrafish embryos were co-exposed to MPs and Cd, the investigators found that low concentrations of MPs (0.05, 0.1 mg/L) showed antagonistic effects on physiological indices in zebrafish, while high-concentration MPs (1, 5, 10 mg/L) showed synergistic sublethal toxicity (Zhang et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e); and this is consistent with the present chapter. However, after clams were exposed to 20 ppb ACP+ PS for 14 days, 0.1% and 1% PS promoted the accumulation of As in clams, but a PS of 5% and 10% inhibited clam accumulation of 20 ppb As (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eA). These results are similar to previous studies: when maize seeds were co-exposed to Cd and MPs, it was revealed that a lower concentration of polyethylene particles (0.1%, 1%) (\u003cem\u003ew/w\u003c/em\u003e) promoted the bioavailability of heavy metals, while a higher concentration (10%) (\u003cem\u003ew/w\u003c/em\u003e) inhibited the bioavailability of heavy metals (Wang et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). Therefore, the amount and number of heavy metals taken up by clams may exert differential effects on various concentrations of PS or PS combined with different types of heavy metals.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec9\"\u003e\n \u003ch2\u003e3.2 Effect of adding PS on the absorption of heavy metals by clams over time\u003c/h2\u003e\n \u003cp\u003eClams were co-exposed to 0%, 1%, or 5% PS and 200 ppb As, Cd, and Pb (ACP) The concentrations of As, Cd, and Pb (wet weight) in clams were measured 1, 3, 5, 7, and 14 days after the start of the exposure experiment (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e and Table S2), and we ascertained that regardless of whether or not PS was added, the concentrations of As, Cd, and Pb in the clams were elevated with time, and the heavy-metal absorptive capacity of the clams was As \u0026gt; Cd \u0026gt; Pb (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). The high content of As was related to the elevated background concentration of As in clams, which is consistent with the tendency for increased metal levels in clams with time after a single heavy-metal exposure. \u003cem\u003eRuditapes philippinarum\u003c/em\u003e also showed a time-dependence after exposure to Cd (4 and 40 \u0026micro;g/L) for 70 days. By the middle of the Cd-exposure period, the Cd concentration in clams increased with exposure time, and then gradually decreased during the middle and later stages (Zhao et al. \u003cspan class=\"CitationRef\"\u003e2014\u003c/span\u003e). When \u003cem\u003eRuditapes philippinarum\u003c/em\u003e clams were exposed to Pb (10 and 100 \u0026micro;g/L) for 7 days, the Pb concentrations also increased with time commensurately (Aouini et al. \u003cspan class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eMPs absorb heavy metals from the aquatic environment (Purwiyanto et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). Although the content of some heavy metals in seawater is low, they can still adsorb MPs when they gather to a certain extent (Gao et al. \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e). Also, the longer the exposure time, the higher the concentration of heavy metals bound by MPs in seawater (Gao et al. \u003cspan class=\"CitationRef\"\u003e2019\u003c/span\u003e); and the addition of MPs exhibits a significant effect on the chemical forms of many heavy metals (Yu et al. \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e). According to Holmes et al., metal adsorption is conducted through the interaction between divalent cations such as Cu\u003csup\u003e2+\u003c/sup\u003e, Pb\u003csup\u003e2+\u003c/sup\u003e, and Cr\u003csup\u003e2+\u003c/sup\u003e, and oxygen anions such as Cr\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e2\u0026minus;\u003c/sup\u003e that possess charged or polar regions on the plastic surface; and via the nonspecific interactions between neutral metal-organic complexes and the hydrophobic surface of bulk plastic medium (Holmes et al. \u003cspan class=\"CitationRef\"\u003e2012\u003c/span\u003e). When the interaction between MPs and heavy metals is via physical adsorption, the chemical bond is weaker and the metals can be more easily released into the aquatic ecosystem again (Purwiyanto et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). However, PS adsorb Cu and Cd principally by chemical adsorption (Xu et al. \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec10\"\u003e\n \u003ch2\u003e3.3 Promotion or inhibition of metal accumulation in clams due to PS\u003c/h2\u003e\n \u003cp\u003eClams exposed to 200 ppb ACP and 1% PS exhibited augmented bioaccumulation of As on days 1, 3, and 5, but showed inhibition on days 7 and 14\u0026mdash; indicating that ACP gradually inhibited 20-ppb As absorption with time and 5% PS inhibited As accumulation within 14 days (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Adding 1% or 5% of PS generally inhibited Cd accumulation in clams over time, and only 5% PS promoted Cd accumulation in clams on the 14th day (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). The addition of 1% PS promoted Pb uptake on the first day, but inhibited it from 3 to 14 days, and the addition of 5% PS stimulated the uptake of Pb (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eThe bioaccumulation of heavy metals in clams due to MPs should be observed over a range of times because a single timepoint produces insufficient and potentially inaccurate data. Overall, 1% PS inhibited the accumulation of Cd and Pb within 14 days and gradually inhibited the uptake of As; while 5% PS inhibited the absorption of As and Cd and promoted the bioaccumulation of Pb within 14 days (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec11\"\u003e\n \u003ch2\u003e3.4 Correlation analysis\u003c/h2\u003e\n \u003cp\u003eUsing correlation analysis, we found that there was also a correlation between the concentration of heavy metals in clams and the concentration of MPs in seawater. At low concentrations of PS plus 20 ppb ACP, As concentration was positively correlated with PS concentration in clams (r\u003csup\u003e2\u003c/sup\u003e = 0.335, P \u0026lt; 0.05) (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA); however, Cd and Pb were not correlated with PS concentration (Fig. 3CE). Simultaneous exposure to low-concentration PS and 200 ppb ACP resulted in a negative correlation between the concentration of PS and concentrations of As and Cd in clams (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eAC), although there was no correlation with the concentration of Pb (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eE). These data thus illustrated an inverse relationship between As/Cd and PS\u0026mdash;i.e., when clams were exposed to 200 ppb As or Cd and a concentration of PS below 1% (\u003cem\u003ew/w\u003c/em\u003e), we observed that a higher concentration of PS was concomitant with a lower concentration of As or Cd. When exposed to high-concentration PS and 20 ppb ACP, the concentrations of Cd and Pb in clams were significantly negatively correlated with the concentration of PS in seawater(r\u003csub\u003eD\u003c/sub\u003e\u003csup\u003e2\u003c/sup\u003e=0.725, p\u0026lt;0.05) (r\u003csub\u003eF\u003c/sub\u003e\u003csup\u003e2\u003c/sup\u003e=0.647, p\u0026lt;0.05) (Fig.\u0026nbsp;3DF); i.e., when clams were co-exposed to high concentration PS (5\u0026ndash;10%) and 20 ppb Cd and Pb, the concentration of Cd and Pb diminished with increasing PS concentration.\u003c/p\u003e\n \u003cp\u003eTable 2 depicts the correlation between a series of PS concentrations and the bioaccumulation of heavy-metal concentrations in clams. We observed a negative correlation between the concentration of PS and 20 or 200 ppb As (r = \u0026minus;0.237, p \u0026gt; 0.05; r = \u0026minus;0.718, p \u0026lt; 0.05), but the concentration of PS was positively correlated with the 20 and 200 ppb Cd (r =0.102, p \u0026gt; 0.05; r =0.341, p \u0026gt; 0.05). Also, the concentration of PS was negatively correlated with the 20 ppb Pb (r = -0.331, p \u0026gt; 0.05) and 200 ppb Pb (r = 0.038, p \u0026gt; 0.05) in clams.\u003c/p\u003e\n \u003cp\u003ePS can affect the metabolic responses of mussels, but the effect is small. MPs can bioaccumulate in shellfish (oysters)\u0026mdash;usually in the form of fibers in gills and mantle\u0026mdash;and they can be involved in the adsorption and precipitation of trace metals in oysters. Researchers also found that the in vivo concentration of MPs was correlated with the in vivo concentrations of Cd, chromium (Cr), Cu, and Pb, indicating that there exists a potential in-vivo interaction between MPs and trace metals (Zhu et al. \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). However, the correlations between the concentration of MPs in water and the concentrations of MPs in organisms, and between the concentrations of heavy metals bound to MPs in organisms and the concentration of heavy metals in the organisms have not been fully elucidated and thus require further investigation.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Conclusion","content":"\u003cp\u003ePS affected the accumulation of As, Cd, and Pb in \u003cem\u003eRuditapes philippinarum\u003c/em\u003e. Different concentrations of PS exerted different effects on the absorption of heavy metals by clams. A low concentration of PS (0.1%, 1%) inhibited heavy-metals (As, Cd, and Pb) bioaccumulation, while a high concentration of PS (5%, 10%) promoted their accumulation. Whether or not PS was added, the concentration of As, Cd, and Pb in clams increased with time. The relative amount bioaccumulated in clams was As \u0026gt; Cd \u0026gt; Pb. The bioaccumulation of heavy metals in clams due to MPs should be observed over a range of times because a single timepoint produces insufficient and potentially inaccurate data. 1% PS inhibited the accumulation of Cd and Pb within 14 days and gradually inhibited the uptake of As; while 5% PS inhibited the absorption of As and Cd and promoted the bioaccumulation of Pb within 14 days. When exposed to a low concentration of PS, the concentrations of As and Cd in clams diminished with the concentration of PS in sea water; and when exposed to high concentrations of PS, the concentrations of Cd and Pb in clams appeared also to be reduced with increasing PS concentrations.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u0026nbsp;\u003c/strong\u003eWe would like to thank Prof. Yuyu Jia and Deyi Wang for his constructive comments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contribution\u0026nbsp;\u003c/strong\u003eThe author Zhen Gao and Zhe Sun were mainly responsible for experimental research, and authors of Yuyu Jia and Deyi Wang were mainly responsible for coaching.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003eThis work was financially supported by the National Natural Science Foundation of China (Grant No. 31900938), the Natural Science Foundation of Shandong Province (Grant No. ZR2019BB073).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability:\u003c/strong\u003e Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e: Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e: Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e: The authors declare no conflict of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAlimi OS, Budarz JF, Hernandez LM, Tufenkji N (2018) Microplastics and Nanoplastics in Aquatic Environments: Aggregation, Deposition, and Enhanced Contaminant Transport. 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Bull Environ Contam Toxicol 107:665\u0026ndash;670. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00128-020-02866-1\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTable.1 The mean concentration of heavy metals in \u003cem\u003eRuditapes philippinarums\u003c/em\u003eafter co-exposure of different concentrations of PS\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.463414634146343%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"27.84552845528455%\"\u003e\n \u003cp\u003eAs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"27.84552845528455%\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"27.84552845528455%\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.56441717791411%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1% PS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e5% PS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1% PS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e5% PS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1% PS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e5% PS\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.56441717791411%\"\u003e\n \u003cp\u003e1 d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.12\u0026plusmn;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.74\u0026plusmn;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.85\u0026plusmn;0.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.95\u0026plusmn;0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.33\u0026plusmn;0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.21\u0026plusmn;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.56441717791411%\"\u003e\n \u003cp\u003e3 d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.10\u0026plusmn;0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.92\u0026plusmn;0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.57\u0026plusmn;0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.63\u0026plusmn;0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.71\u0026plusmn;0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.10\u0026plusmn;1.11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.56441717791411%\"\u003e\n \u003cp\u003e5d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.12\u0026plusmn;0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.68\u0026plusmn;0.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.77\u0026plusmn;0.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n 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\u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.74\u0026plusmn;0.18\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.56441717791411%\"\u003e\n \u003cp\u003e14d\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.50\u0026plusmn;0.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.76\u0026plusmn;0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.71\u0026plusmn;0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.25\u0026plusmn;0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e0.77\u0026plusmn;0.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"13.905930470347649%\"\u003e\n \u003cp\u003e1.06\u0026plusmn;0.33\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003eTable.2 Pearson correlation between PS and heavy metal in \u003cem\u003eRuditapes philippinarums\u003c/em\u003e\u003c/p\u003e\n\u003cdiv align=\"center\" id=\"isPasted\"\u003e\n \u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003eAs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003eCd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003ePb\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e20ppb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e200ppb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e20ppb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e200ppb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e20ppb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e200ppb\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003ePS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-0.237\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-0.718\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.102\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.341\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-0.331\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.038\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Clam, Arsenic, Cadmium, Lead, Microplastics, Bioaccumulation","lastPublishedDoi":"10.21203/rs.3.rs-1237924/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1237924/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMicroplastics constitute a new type of environmental pollutant with a particle size of less than 5 mm that can combine with heavy metals to enter organisms such as clams, and the impact of the combination of microplastics and heavy metals on the marine organism \u003cem\u003eRuditapes philippinarum\u003c/em\u003e remains unclear. Therefore, this problem is analyzed the effects of different concentrations (0, 0.1, 1, 5, and 10% \u003cem\u003ew/w\u003c/em\u003e) of polystyrene (PS) and three heavy metals (arsenic (As), cadmium (Cd), and lead (Pb)) (20 and 200 \u0026micro;g/L) on the bioaccumulation of the heavy metals in clams, and the effects of time for PS (0, 1, and 5%)-bound As, Cd, and Pb (ACP). We found that different concentrations of PS exerted different effects on the absorption of heavy metals by clams. A low concentration of PS (0.1%, 1%) inhibited heavy-metal bioaccumulation, while a high concentration of PS (5%, 10%) promoted their accumulation. Whether or not PS was added, the concentration of As, Cd, and Pb in clams increased with time. The relative amount bioaccumulated in clams was As \u0026gt; Cd \u0026gt; Pb, and we demonstrated a correlation between the concentration of heavy metals in clams and the concentration of PS in seawater. When exposed to a low concentration of PS, the concentrations of As and Cd in clams diminished with the concentration of PS in sea water; and when exposed to high concentrations of PS, the concentrations of Cd and Pb in clams appeared also to be reduced with increasing PS concentrations.\u003c/p\u003e","manuscriptTitle":"Effects of Different Polystyrene Concentrations on the Bioaccumulation of As, Cd, and Pb in Ruditapes Philippinarum","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-02-15 23:18:03","doi":"10.21203/rs.3.rs-1237924/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"54d2541c-1b11-4f00-9a81-55483d51ce89","owner":[],"postedDate":"February 15th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2022-05-19T04:40:50+00:00","versionOfRecord":[],"versionCreatedAt":"2022-02-15 23:18:03","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1237924","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1237924","identity":"rs-1237924","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}