Next Article in Journal
Integrated Planning of Feeder Route Selection, Schedule Design, and Fleet Allocation with Multimodal Transport Path Selection Considered
Next Article in Special Issue
Real-Time Instance Segmentation for Detection of Underwater Litter as a Plastic Source
Previous Article in Journal
Size Selectivity of a Diamond-Mesh Codend of Demersal Trawl for Largehead Hairtail (Trichiurus lepturus Linnaeus, 1758) in the Beibu Gulf, in the South China Sea
Previous Article in Special Issue
Floating Riverine Litter Flux to the White Sea: Seasonal Changes in Abundance and Composition
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
JMSE
Volume 11
Issue 7
10.3390/jmse11071446
jmse-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Biomarker Effects of Diesel Fuel Hydrocarbons Absorbed to PE-Plastic Debris on Mussel Mytilus trossulus

by
Nadezda Vladimirovna Dovzhenko
,
Victor Pavlovich Chelomin
,
Andrey Alexandrovich Mazur
,
Valentina Vladimirovna Slobodskova
,
Aleksandra Anatolyevna Istomina
and
Sergey Petrovich Kukla
*
V.I. Il’ichev Pacific Oceanological Institute, Far Eastern Branch, Russian Academy of Sciences, 690041 Vladivostok, Russia
*
Author to whom correspondence should be addressed.
J. Mar. Sci. Eng. 2023, 11(7), 1446; https://doi.org/10.3390/jmse11071446
Submission received: 30 June 2023 / Revised: 12 July 2023 / Accepted: 18 July 2023 / Published: 19 July 2023
(This article belongs to the Special Issue Marine Litter and Sustainability of Ocean Ecosystems)
Browse Figures
Versions Notes

Abstract

:
Pollution of global oceans by plastic litter is one of the most important ecological problems of our time. At the same time, the active sorption of highly toxic chemicals dissolved in water by plastic also poses a threat to the marine environment and its inhabitants. This article presents the results of experimental studies on the properties of polyethylene (PE) as a vector of petroleum hydrocarbons (PH) and its influence on the biochemical parameters of tissues in the Pacific mussel Mytilus trossulus. It was shown that the presence of unused polyethylene fragments (PE) and PE fragments with water-soluble fractions of diesel fuel (PE-WSF-DF) in seawater caused the development of oxidative stress in M. trossulus. We observed severe changes in hemolymph lysosome membrane stability (LMS) and a significant increase in DNA fragmentation in the gills and digestive glands of mollusks. The presence of PE-WSF-DF fragments in water increased the activity of antioxidant enzymes: catalase (CAT) and glutathione-S-transferase (GST). In the experiment, direct contact between plastic fragments and mussels was excluded, meaning the filter-feeding mollusks ingested the desorbed PH and leaching components existing in PE from the water.

1. Introduction

The most significant consequences of various forms of industrial and domestic human activities are manifested in coastal marine ecosystems, especially in enclosed and semi-enclosed bays, marine lagoons, and river estuaries. A great variety of pollutants are constantly introduced and concentrated in these areas by numerous river and terrigenous runoffs, wind drifts, and coastal currents. Heavy metals (HM), pesticides, and petroleum hydrocarbons (PH) account for most of these substances. In recent years, these “traditional” pollutants have been joined by plastic debris, which forms huge accumulations of plastic products and their fragments in the tidal zone around the world [1,2,3,4]. Plastic is already confidently considered one of the fastest-growing components polluting coastal ecosystems, given the current nature of man-made polymer production, where single-use products dominate and more than 50% of the world’s inhabitants live within a 50-mile zone along the marginal seas.
Plastic fragments, due to their hydrophobic properties, can adsorb various persistent and highly toxic pollutants, such as HM, pesticides, and polycyclic aromatic hydrocarbons [5,6,7,8,9,10,11]. These exogenous low molecular weight chemical compounds are physically bound to polymeric structures and can be released relatively freely into the environment under appropriate conditions. Even though the ratio of adsorption and desorption processes of these components depends on factors such as the concentration and chemical properties of polymers and toxicants, hydrochemical characteristics of the environment (temperature, pH, and salinity) [12,13,14,15,16,17], the plastic circulating in these coastal areas together with other hazardous chemicals represents a serious ecotoxicological threat to various marine organisms. Depending on their size, plastic fragments penetrate into the digestive system of relatively large marine species by ingestion, where the sorbed pollutants are desorbed, along with the corresponding harmful effects [18,19,20,21]. Plastic fragments saturated with sorbed toxic components in contaminated coastal areas can be transported relatively quickly over long distances under the influence of wind and coastal currents. In unpolluted waters or in waters with other hydrochemical characteristics, most contaminants can desorb from plastic fragments and threaten a wider range of aquatic organisms, including small-sized organisms and their planktonic forms [6].
From the ecotoxicological point of view, the development of these negative events seems most real for the inhabitants of the littoral–sublittoral zone, which is characterized by sharp changes in hydrochemical parameters, such as temperature, salinity, pH, and concentrations of dissolved oxygen and carbon dioxide in the water. We believe that within the context of the problem under consideration, model experiments involving macroplastics, and to a certain extent filter organisms, can simulate the real situation periodically occurring in the littoral zone and provide the most convincing assessment of the danger caused by “contaminated” plastic in marine ecosystems.
To show the ability of plastic to sorb and transfer hazardous chemicals to aquatic organisms through the water column, we used a water-soluble fraction of diesel fuel (WSF-DF) as a pollutant, to represent an oil refinery product. Today, petroleum hydrocarbons are still a major pollutant in coastal waters, especially in semi-enclosed bays with a massive accumulation of plastic debris, and they constantly attract increased attention from ecotoxicologists because of their potentially harmful effects on marine organisms [22,23,24,25].
The aim of this paper was to study the effect of polyethylene macrofragments, which had been saturated with diesel fuel hydrocarbons, on biochemical markers of oxidative stress (lysosomal membrane stability (LMS), DNA damage, total oxyradical scavenging capacity (TOSC), lipid peroxidation products (malonaldehyde (MDA)), and activity of antioxidant enzymes) in the Pacific mussel Mytilus trossulus. This littoral bivalve mollusk has a high filtration rate and is a typical representative of coastal ecosystems in the Sea of Japan. In addition, it is prone to bioaccumulation of xenobiotics and shows high sensitivity to various chemical stressors [26].

2. Materials and Methods

2.1. Description of the Experiment

The study was carried out at the marine experimental station “Popov Island” of the V.I. Il’ichev Pacific Oceanological Institute. The mussels M. trossulus (Gould, 1850) were collected by SCUBA divers from natural populations in Alexeev Bay (the Peter the Great Bay, Sea of Japan).
Mussels that were in the post-spawning period, 4 ± 0.5 cm in size were selected for the experiment. The mussels were pre-acclimated in laboratory conditions for 7 days. Mussels were kept in a glass tank with forced aeration at a water temperature of +16 °C and water salinity of 32.2–32.6 psu. During the acclimation and experiments, the mussels were not fed.
Experiments with mollusks were carried out in three tanks at a stable water temperature of +16 °C. The volume of seawater in each tank was 25 L (0.5 L for each mollusk). The water in the tanks was intensively aerated to ensure high oxygen content. In addition, this procedure created a simulation of currents and wave motion in the tanks that promoted the leaching of endogenous and exogenous chemicals from the experimental polyethylene (PE) fragments. The exposure period was 72 h.
In the first series of experiments, “virgin” (unused) fragments of low-pressure polyethylene (PE) film were placed in the aquariums with the experimental mollusks (group 1). The film (produced in Russia, State standard PE-16338-85) was cut into 10 × 10 cm fragments and used in this study. The total area of plastic fragments for each experimental tank was 0.04 m2/L.
In the second series of experiments, mussels (experimental group 2) were kept in the presence of PE fragments preincubated in a water-soluble fraction of diesel fuel (WSF-DF). The WSF-DF solution was prepared using the standard method in a 1:10 ratio, to represent the basis of summer diesel fuel and filtered seawater [25].
The composition of WSF-DF was previously investigated using the chromato–mass spectrophotometric method [25]. The concentration of water-soluble hydrocarbons was 10 mg/L. The 10 × 10 cm PE fragments were incubated for 24 h in the prepared WSF-DF solution with constant shaking. After incubation in the solution, fragments with sorbed hydrocarbons were removed and air-dried at +20 °C for 5 h. Then, the prepared PE-WSF-DF fragments (0.04 m2/L) were placed in a glass tank with the mollusks in experimental group 2. The control group of mussels was placed in the third tank.
The control group included mussels that were kept in a separate tank under the same conditions but without the addition of any plastic fragments.

2.2. Determination of Biochemical Parameters

Lysosomal membrane stability in mollusk hemocytes was determined using the method proposed by Martínez-Gomez et al. [27]. Hemolymph from mollusks was collected using a 0.1 mL hypodermic syringe from the muscle locker. Twenty samples (n = 20) were taken from each experimental group for analysis. The incubation time for each sample in the dye was 15 min. Samples were viewed under a microscope at 15 min intervals for 90 min.
The digestive gland and gills of each mollusk were used for the analysis of various biochemical markers (enzyme activity, DNA damage, integral antiradical activity index TOSC (total oxyradical scavenging capacity), and malondialdehyde).
To determine enzyme activity, tissues were homogenized in cooled 0.1 M phosphate buffer (pH 7.0) at a ratio of 1:10 g/mL (4 °C). The homogenate was centrifuged at 10,000× g for 40 min at 4 °C. The supernatants were used to determine the catalase (CAT) and glutathione-S-transferase (GST) activities. The activity of CAT was determined spectrophotometrically according to the method of Regoli and Principato [28]. The GST activity was determined according to the method described by Habig et al. [29]. The activities of the enzymes are expressed in μmol/min/mg protein. Determination of the total oxyradical scavenging capacity (TOSC) in the tissue supernatant was performed using the method previously described by Bartosz et al. [30]. The activity value was calculated using a calibration plot constructed using Trolox (6-hydroxy-2,5,7,8-tetramethylchloraman-2-carboxylic acid, Sigma Aldrich, Moscow, Russian Federation). The content of malonic dialdehyde in the molluscan tissues was determined spectrophotometrically by reacting with thiobarbituric acid (TBA) [31]. The protein concentration in the samples was determined using the modified Lowry method [32].
An alkaline version of the Comet assay [33], adapted to marine organisms [34], was used to determine the degree of DNA molecule damage. Casp 1.2.2 software (CASPlab, Wroclaw, Poland) was used for digital image processing.

2.3. Statistical Analysis

Statistical processing of the results was performed using MS Office Excel statistical tools and Statistica software package. The significant differences between the samples were determined by the nonparametric Mann–Whitney test. Differences were considered statistically significant at p < 0.05.

3. Results

Under our experimental conditions, the presence of “virgin” (unused) polyethylene (PE) macrofragments and polyethylene sorbing in the water of water-soluble hydrocarbons from diesel fuel (PE-WSF-DF) had no effect on the survival of the mussels in both experimental groups (groups 1 and 2). At the same time, these mussels showed various sublethal effects on the biochemical marker levels.

3.1. Lysosomal Membrane Stability (LMS)

Figure 1 shows the results of a series of experiments demonstrating the effect of PE and PE-WSF-DF fragments on lysosomal membrane stability in the hemocytes of the marine bivalve mollusk M. trossulus.
The presence of both types of plastics in the tanks caused serious changes in the stability of the lysosome membranes. In the experimental mussels (group 1), after adding “virgin” PE fragments, the hemocyte LMS decreased practically 2.5-fold compared to the mollusk control group. A similar effect was shown in group 2 when PE-SF-DF fragments were present in the tank.

3.2. Total Oxyradical Scavenging Capacity (TOSC)

The susceptibility of the digestive gland and gill cells in the experimental mussels to oxidative stress was assessed by analyzing the total ability of the low molecular weight component of the AO system of mussels (TOSC) to neutralize the radical (ROO-). Figure 2 shows the TOSC levels measured in the control group mussel tissues and in mussels from experimental groups 1 and 2, which were kept in tanks with PE and PE-WSF-DF fragments, respectively.
According to the data, the reaction by the experimental mollusks to the presence of both types of plastic in the medium was tissue-specific. No significant changes were shown in the TOSC in the digestive gland. Whereas in the gill cells, this index sharply decreased by more than 40% in both experimental groups compared to the control group, which indicates the suppression of the low-molecular-weight component in the AO system in the presence of the studied types of plastic.

3.3. Products of Peroxidation (MDA)

According to the results shown in Figure 3, no significant differences were observed in the level of TBA-active products in the experimental mussel tissues compared to the control group. These results indicate the absence of the accumulation of these reactive products in the cell membrane lipid matrix.

3.4. Enzyme Activity

The results of the activity of the AO enzymes in the tissues of all the experimental mussel groups are shown in Figure 4.
The presence of plastic in the tanks of the experimental mussels caused an increase in CAT activity in the cells of the digestive gland but had no effect on the activity of this enzyme in the gill cells. Although CAT activity increased in the digestive gland in both experimental groups of mollusks, significant differences were observed only between the control mussels and those in group 2 (Figure 4A). In contrast to CAT, the activity of the GST was increased in both the digestive gland and gills of the experimental mollusks. Nevertheless, as in the case of CAT, significant differences were only observed between the control mussels and mussels exposed to PE-WSF-DF fragments (Figure 4B).

3.5. Damage to DNA Structure

Comet analysis provided convincing evidence that exposure to both types of plastic (PE and PE-WSF-DF) causes genotoxic effects to varying degrees in the tissues of M. trossulus (Figure 5). Figure 5 shows one of the quantitative parameters of the comets obtained by this method (averaged values of DNA percentage in the “tail” of comets), characterizing the degree of damage to the nuclear DNA molecule of the digestive gland cells and gills of the control and experimental mollusks.
A significant increase in the proportion of fragmented DNA migrating from the nucleus to the comet “tail” is common in the mussel experimental groups. The data presented in Figure 5 shows that in mussels where only fragments of “virgin” PE were present in the tank, the level of DNA damage in the digestive gland and gill cells increased, almost by 1.5- and 2-fold, respectively, compared to the control mollusks. In the group 2 experimental mussels, after exposure to the PE-WSF-DF fragments, the process of destruction significantly increased and the level of fragmented nuclear DNA in the cells of both tissues increased by almost 3-fold compared to the control mollusks.

4. Discussion

All modern types of plastic are a complex mixture of chemical compounds based on a high-polymer structure with various low molecular weight chemical additives. Additives are added during polymer synthesis to carry out the polymerization process (catalysts, stabilizers, etc.), and provide the plastic-specific physical properties (for example, HM, bisphenol A, phthalates, polychlorinated biphenyls, dyes, etc.) necessary for use in the national economy [35,36]. As a rule, the polymerization process does not proceed completely, which leads to the presence of unreacted monomers, oligomers, and reaction byproducts in the composition of plastics, the amount of which varies over a wide range [36,37]. Additionally, these endogenous chemical compounds and synthetic polymers can sorb a variety of chemicals through their surface due to their hydrophobic properties, which they interact with during their use, as well as along their migratory paths, both on land and in the aquatic environment.
Endogenous and exogenous chemical compounds are very diverse in nature and properties, although a common property is that they can bind to the structures of the plastic but not to chemical (not covalent) bonds, although only by relatively weak physical bonds. Some of the chemical substances may leach out of the plastic and into the environment, causing the marine organism’s exposure or external conditions (physical and chemical characteristics) to change [38,39,40,41].
In this respect, a dangerous situation occurs in the coastal belt of marginal seas, where huge masses of plastic debris accumulate from various sources, mainly winds, coastal currents, and river runoffs [1,2,3,4]. Plastic debris located in the shoreline and intertidal zone is exposed to numerous external factors, such as mechanical rubbing with sand and small rocks, high water dynamics, fluctuations in temperature, salinity, pH, abundance of oxygen, and ultraviolet light. The combination of these factors and the abundance of plastic trash in this “transit” area makes it necessary not to neglect the potential hazard to which the organisms living there are exposed. It is practically impossible to reproduce the effect of all these factors on the leaching of endogenous and sorbed chemical compounds and to estimate their integral concentration under laboratory conditions.
Therefore, in short-term experiments, we approximately simulated real conditions of cohabitation by the filter mollusk M. trossulus with fragments of “virgin” polyethylene, excluding chemical sorption and polyethylene sorbing WSF-DF, followed by an assessment of the consequences and effects of these polymers by using oxidative stress molecular markers.
The reaction of two markers, LMS and tissue cell genome degradation, which showed high sensitivity to both types of plastic under the conditions of our short-term experiments, should be especially emphasized.
A sharp decrease in the stability of the hemocyte lysosome membranes in group 1 and, especially, group 2 mussels can be considered a consequence of a general nonspecific reaction to chemical ingestion and can be explained by a position of generally accepted notions—an increased generation of oxyradicals [42,43,44]. These assumptions are based on the well-known functions of lysosomes and literature data, which show that various components of oil and chemicals can leach from artificial polymers and cause a reduction in the stability of the lysosome membranes in bivalve mollusks [22,38,45,46,47].
The high sensitivity of LMS was demonstrated by Avio et al. [43], whereby the exposure of mussels to microplastic led to a decrease in LMS, despite weak changes in TOSC and MDA. The destabilization of the lysosome membrane can consistently lead to severe dysfunction in cells and tissues, compromising cell viability or initiating apoptosis [48,49]. According to the comet assay results shown in Figure 5, the common thing for both mussel experimental groups was a significant increase in the proportion of fragmented DNA in the gill and digestive gland cells migrating from the nucleus to the comet tail. We assume that chemicals leached from plastic fragments during the experiments caused genome destabilization. It is well known that a wide range of chemical compounds exhibit genotoxic properties, including both monomers of the various types of polymers and chemical additives used in their synthesis, and from the hydrocarbons of the oil and diesel fuel being transferred into the aqueous phase [23,50,51,52,53]. Although the PE monomers we used in our experiments are classified as “low-hazard” [54] or “of concern” [55], the chemical additives in this plastic exhibit genotoxicity. An especially high degree of DNA destruction was observed in the gill and digestive gland cells of the group 2 experimental mussels, which were kept in the presence of PE-WRF-DF. We previously showed [25] that the water-soluble diesel fuel fraction (WRF-DF), which we used in our experiments with the group 2 mussels, contains chemical components that enhance nuclear DNA damage. Genotoxicity is caused by the presence of toxic mono-, di-, and polyaromatic hydrocarbons in this fraction, which account for about 20% [25]. In this regard, our ideas are consistent with data from numerous previous studies showing that water-soluble hydrocarbons from oil and diesel fuel cause DNA damage in the cells of various hydrobionts [22,23,24]. These hydrocarbons, which are part of the water-soluble fraction of diesel fuel, can induce highly reactive oxyradicals, which are the main cause of DNA oxidative damage and the destabilization of lysosome membranes [24,52].
It is noteworthy that there are no significant changes in the MDA content in the tissues of the experimental mollusks compared to the control group (Figure 3), which is consistent with the data by Nogueira et al. [56]. In their study, after exposure to the bivalve Perna perna from biodiesel hydrocarbons, there was also no increase in MDA levels in tissues against a background of decreased activity of antioxidant enzymes. Although this index is widely used as a marker of oxidative stress in bivalves [57], there is a possibility that it is less sensitive under short-term exposure to low concentrations of toxic chemicals [58,59]. This assumption is based on the increased resistance to pro-oxidative processes by mussel cell membrane lipids, the composition of which revealed a high content of weakly oxidizable non-methylene-separated fatty acids [60]. In addition, it is logical to assume that chemicals leached from the plastic can induce the activity of glutathione peroxidase and GST enzymes in the cells of mussels, which are enzymes directly involved in the detoxification of fatty acid hydroperoxides and nucleophilic carbonyls [61]. According to our experimental data (Figure 4B), in the presence of both types of plastic, the activity of the GST enzyme increased to different degrees in the mussel tissues. Our results indicate that the response of biomarkers, such as the TOSC level and the activities of the CAT and GST enzymes, was tissue-specific and depended on the type of experimental plastic being used (PE or PE-WRF-DF).
The assimilation of nutrient organic and inorganic substances dissolved in seawater takes place through the gills. These substances then enter the hemolymph, which transports them to other organs. For this reason, during the filtration process, the chemicals leached from the plastic can have a direct effect, initially, on the biochemical system of the gill cells. Then, probably already in a weakened form, their influence is exerted on other mollusk organs and tissues, which primarily begin in the cells of the digestive gland.
In the presence of “virgin” and contaminated PE in mollusks, a decrease in TOSC level was observed in the gill cells (Figure 2), which indicated an increased generation of oxyradicals and the development of pro-oxygenic processes. In the cells in the digestive gland, a slight increase in antiradical activity (TOSC) was observed under the influence of the “virgin” plastic, which was obviously associated with the activation of compensatory mechanisms. These results support our view that in mollusks the antioxidant system in the digestive gland cells is more efficient compared to in the gill cells [59,62].
The probability of chemicals leaching from plastic into the water column and their penetration into the mollusk body can be estimated by changes in the activity of the glutathione-s-transferase (GST) enzyme. GST is a biomarker of exposure to a wide range of organic pollutants because it participates in the detoxification of xenobiotics, including petroleum hydrocarbons, in Phase II of metabolic transformations [58,59,63].
According to the data presented in Figure 4B, we can state that GST activity is induced in both mussel experimental groups. An increase in GST activity was recorded in the tissues of the group 2 mussels (PE-WSF-DF), indicating partial desorption of diesel hydrocarbons from PE-WSF-DF fragments and their penetration into the mussel tissues. A similar response was observed for glutathione-S-transferase to short-term exposure to different concentrations of WSF-DF in polychaetes (Laeonereis culveri), paddlefish (copepods Calanus glacialis, Calanus finmarchicus) and the bivalve clam Anomalocardia flexuosa [24,64]. A significant increase in glutathione-S-transferase activity was shown in the digestive gland of the mussel Crenomytilus grayanus in areas with the highest oil-hydrocarbon content in soils compared to clams from a conditionally clean area [58]. The work of Pandi et al. [63] can be cited as an indirect confirmation of our assumptions. The authors showed that the weathered micro-PE modulates the activity and enhances the expression of GST genes in Danio albolineatus fish.
We recorded a weak increase in CAT activity in only the digestive gland cells of the experimental animals in the presence of the PE-WSF-DF fragments (Figure 4A). Nogueira et al. [56] came to a similar conclusion based on the lack of the response by this enzyme in the clam Perna perna, even after long-term exposure to biodiesel (biodiesel B5). Apparently, catalase is not the main link in the enzymatic antioxidant system in the mussel M. trossulus in the detoxification of hydrogen peroxide, which is confirmed by its low activity among marine bivalves [26].
Our experiments involved large fragments of plastic and immobile filter organisms, which excludes their direct contact, although it suggests the effect of chemicals desorbed from PE through the water column. We did not analyze the types and concentrations of the chemical compounds leaching into the water during the experiments; however, from the reaction of the biomarkers, we can state with a high probability that biochemical changes in the mussel tissues were initiated by endogenous (experimental group 1) and exogenous (experimental group 2) chemicals leached from the plastic.
It is important to emphasize that regardless of the chemical substances leached from the PE and PE-WSF-DF fragments, negative effects were observed when M. trossulus were briefly exposed to them, despite the developed xenobiotic detoxification system in these mollusks.
Given that the leaching rate of specific additives or desorption of water-soluble hydrocarbons of diesel fuel from plastic is low, given the huge scale of accumulation of fragments of artificial polymers in the coastal zone, the concentration of these toxic substances in local areas may be sufficient to cause biological effects. The risk of various biological effects increases further when noting that the littoral zone is home to invertebrates prone to the bioaccumulation of various xenobiotics.
Biomarkers are commonly used to predict changes in higher biological organization levels and are defined as “short-term indicators of long-term biological effects” [65]. Thus, chronologically related disturbances in biochemical processes, which are signaled by the changes in the markers shown in our work, can be an early stage in the development of pathological events that lead to a lethal outcome. We believe that the biochemical changes identified in the work, based on the biomarker reactions leading to oxidative stress and genotoxicity, do not exhaust the whole range of consequences, which is obviously much wider. Nevertheless, genotoxicity, which is critical and carries serious risks for the future, should be highlighted. Taking into account that the comet assay method only assesses changes in the nucleus in the early stages of genome (DNA) destruction development, the genotoxic properties of chemicals leached from both types of plastics revealed in our experiments are prognostic in nature.
Overall, the experimental results revealed an additional potential risk problem arising from plastic pollution in marine coastal ecosystems.

Author Contributions

Conceptualization, V.P.C. and N.V.D.; methodology, V.V.S. and A.A.I.; soft-ware, S.P.K.; validation, A.A.M. and V.V.S.; formal analysis, A.A.M. and S.P.K.; investigation, A.A.I., V.V.S. and S.P.K.; resources, V.P.C.; data curation, N.V.D. and A.A.I.; writing—original draft preparation, N.V.D. and V.P.C.; writing—review and editing, N.V.D., A.A.M., and S.P.K.; visualization, S.P.K.; supervision, V.P.C. and N.V.D.; project administration, V.P.C.; funding acquisition, V.P.C. All authors have read and agreed to the published version of the manuscript.

Funding

This research was supported by the state assignment for research work of V.I. Il’ichev Pacific Oceanological Institute, FEB RAS (No. 121021500052-9).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Galgani, F.; Leaute, J.P.; Moguedet, P.; Souplet, A.; Verin, Y.; Carpentier, A.; Goraguer, H.; Latrouite, D.; Andral, B.; Cadiou, Y.; et al. Litter on the sea floor along European coasts. Mar. Pollut. Bull. 2000, 40, 516–527. [Google Scholar] [CrossRef]
  2. Kako, S.; Isobe, A.; Kataoka, T.; Hinata, H. A decadal prediction of the quantity of plastic marine debris littered on beaches of the East Asian marginal seas. Mar. Pollut. Bull. 2014, 81, 174–184. [Google Scholar] [CrossRef]
  3. Iñiguez, M.E.; Conesa, J.A.; Fullana, A. Marine debris occurrence and treatment: A review. Renew. Sust. Energ. Rev. 2016, 64, 394–402. [Google Scholar] [CrossRef] [Green Version]
  4. Jang, Y.; Kim, K.N.; Woo, J.R. Post-consumer plastic packaging waste from online food delivery services in South Korea. Waste Manag. 2023, 156, 177–186. [Google Scholar] [CrossRef]
  5. Mato, Y.; Isobe, T.; Takada, H.; Kanehiro, H.; Ohtake, C.; Kaminuma, T. Plastic resin pellets as a transport medium for toxic chemicals in the marine environment. Environ. Sci. Technol. 2001, 35, 318–324. [Google Scholar] [CrossRef]
  6. Teuten, E.L.; Saquing, J.M.; Knappe, D.R.U.; Barlaz, M.A.; Jonsson, S.; Bjorn, A.; Rowland, S.J.; Thompson, R.C.; Galloway, T.S.; Yamashita, R.; et al. Transport and release of chemicals from plastics to the environment and to wildlife. Philos. Trans. Royal Soc. 2009, 364, 2027–2045. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  7. Fries, E.; Zarfl, C. Sorption of polycyclic aromatic hydrocarbons (PAHs) to low and high density polyethylene (PE). Environ. Sci. Pollut. Res. Int. 2012, 19, 1296–1304. [Google Scholar] [CrossRef] [PubMed]
  8. Bakir, A.; Rowland, S.J.; Thompson, R.C. Enhanced desorption of persistent organic 421 pollutants from microplastics under simulated physiological conditions. Environ. Pollut. 2014, 185, 16–23. [Google Scholar] [CrossRef]
  9. Gauquie, J.; Devriese, L.; Robbens, J.; De Witte, B. A qualitative screening and quantitative measurement of organic contaminants on different types of marine plastic debris. Chemosphere 2015, 138, 348–356. [Google Scholar] [CrossRef]
  10. Gao, H.; Yan, C.; Liu, Q.; Ding, W.; Chen, B.; Li, Z. Effects of plastic mulching and plastic residue on agricultural production: A meta-analysis. Sci. Total Environ. 2019, 651, 484–492. [Google Scholar] [CrossRef] [PubMed]
  11. Fred-Ahmadu, O.H.; Bhagwat, G.; Oluyoye, I.; Benson, N.U.; Ayejuyo, O.O.; Palanisami, T. Interaction of chemical contaminants with microplastics: Principles and perspectives. Sci. Total Environ. 2020, 706, 135978. [Google Scholar] [CrossRef]
  12. Hartmann, N.B.; Rist, S.; Bodin, J.; Jensen, L.H.; Schmidt, S.N.; Mayer, P.; Meibom, A.; Baun, A. Microplastics as vectors for environmental contaminants: Exploring sorption, desorption, and transfer to biota. Integr. Environ. Assess. Manag. 2017, 13, 488–493. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  13. Rodrigues, S.M.; Almeida, C.M.R.; Silva, D.; Cunha, J.; Antunes, C.; Freitas, V.; Ramos, S. Microplastic contamination in an urban estuary: Abundance and distribution of microplastics and fish larvae in the Douro estuary. Sci. Total Environ. 2019, 659, 1071–1081. [Google Scholar] [CrossRef]
  14. Mei, W.; Chen, G.; Bao, J.; Song, M.; Li, Y.; Luo, C. Interactions between microplastics and organic compounds in aquatic environments: A mini review. Sci. Total Environ. 2020, 736, 139472. [Google Scholar] [CrossRef]
  15. Menéndez-Pedriza, A.; Jaumot, J. Interaction of environmental pollutants with microplastics: A critical review of sorption factors, bioaccumulation and ecotoxicological effects. Toxics 2020, 8, 40. [Google Scholar] [CrossRef]
  16. Song, J.; Na, J.; An, D.; Jung, J. Role of benzophenone-3 additive in chronic toxicity of polyethylene microplastic fragments to Daphnia magna. Sci. Total Environ. 2021, 800, 149638. [Google Scholar] [CrossRef]
  17. Gao, H.; Liu, Q.; Yan, C.; Mancl, K.; Gong, D.; He, J.; Me, X. Macro-and/or microplastics as an emerging threat effect crop growth and soil health. Resour. Conserv. Recycl. 2022, 186, 106549. [Google Scholar] [CrossRef]
  18. Koelmans, A.A.; Gouin, T.; Thompson, R.; Wallace, N.; Arthur, C. Plastics in the marine environment. Environ Toxicol. Chem. 2014, 33, 5–10. [Google Scholar] [CrossRef]
  19. Hermabessiere, L.; Dehaut, A.; Paul-Pont, I.; Lacroix, C.; Jezequel, R.; Soudant, D.G. Occurrence and effects of plastic additives on marine environments and organisms: A review. Chemosphere 2017, 182, 781–793. [Google Scholar] [CrossRef] [Green Version]
  20. Pedà, C.; Caccamo, L.; Fossi, M.C.; Gai, F.; Andaloro, F.; Genovese, L.; Perdichizzi, A.; Romeo, T.; Maricchiolo, G. Intestinal alterations in European sea bass Dicentrarchus labrax (Linnaeus, 1758) exposed to microplastics: Preliminary results. Environ. Pollut. 2016, 212, 251–256. [Google Scholar] [CrossRef] [PubMed]
  21. Rainieri, S.; Conlledo, N.; Larsen, B.K.; Granby, K.; Barranco, A. Combined effects of microplastics and chemical contaminants on the organ toxicity of zebrafish (Danio rerio). Environ. Res. 2018, 162, 135–143. [Google Scholar] [CrossRef] [PubMed]
  22. Giannapas, M.; Karnis, L.; Dailianis, S. Generation of free radicals in haemocytes of mussels after exposure to low molecular weight PAH components: Immune activation, oxidative and genotoxic effects. Comp. Biochem. Physiol. C Toxicol. Pharmacol. 2012, 155, 182–189. [Google Scholar] [CrossRef] [PubMed]
  23. Weber, L.; Carvalho, L.; Sa, N.; Silva, V.; Beraldini, N.; Souza, V.; Conceica, M. Genotoxic effects of the water-soluble fraction of heavy oil in the brackish/freshwater amphipod Quadrivisio aff. lutzi (Gammaridea) as assessed using the comet assay. Ecotoxicology 2013, 22, 642–655. [Google Scholar] [CrossRef]
  24. Braga, M.A.; Brauko, K.M.; Vicentini, M.; Salgado, L.D.; Silva de Assis, H.C.; Dolatto, R.G.; Grassi, M.T.; Sandrini-Neto, L.; Lana, P.C. Cytotoxicity and enzymatic biomarkers as early indicators of benthic responses to the soluble-fraction of diesel oil. Ecotox. Environ. Safety 2018, 164, 21–31. [Google Scholar] [CrossRef]
  25. Chelomin, V.P.; Slobodskova, V.V.; Kukla, S.P.; Zhuravel, E.V.; Chernyaev, A.P. Genotoxic Effects of Exposure to Water-Soluble Fraction of Diesel Fuel in Sand Dollar Scaphechinus mirabilis Gametes. Toxics 2023, 11, 29. [Google Scholar] [CrossRef] [PubMed]
  26. Istomina, A.; Yelovskaya, O.; Chelomin, V.; Karpenko, A.; Zvyagintsev, A. Antioxidant activity of Far Eastern bivalves in their natural habitat. Mar. Environ. Res. 2021, 169, 105383. [Google Scholar] [CrossRef]
  27. Martinez-Gomez, C.; Bignell, J.; Lowe, D. Lysosoma membrane stability in mussels. ICES Tech. Mar. Environ. Sci. 2015, 56, 41. [Google Scholar] [CrossRef]
  28. Regoli, F.; Principato, G. Glutathione, glutathione-dependant and antioxidant enzymes in mussel, Mytilus galloprovincilis exposed to metals in different field and laboratory condition. Implications for a proper use of biochemical markers. Aquat. Toxicol. 1995, 31, 143–164. [Google Scholar] [CrossRef]
  29. Habig, W.H.; Jakoby, W.B. Assays for differentiation of glutathione S-transferases. Methods. Enzymol. 1981, 77, 398–405. [Google Scholar] [CrossRef]
  30. Bartosz, G.; Janaszewska, A.; Ertel, D.; Bartosz, M. Simple determination of peroxyl radical-trapping capacity. Int. J. Biochem. Mol. Biol. 1998, 46, 519–528. [Google Scholar] [CrossRef]
  31. Buege, J.A.; Aust, S.D. Microsomal lipid peroxidation. Methods Enzymol. 1978, 52, 302–310. [Google Scholar] [CrossRef]
  32. Markwell, M.A.; Haas, S.M.; Bieber, L.L.; Tolbert, N.E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal. Biochem. 1978, 87, 206–210. [Google Scholar] [CrossRef]
  33. Singh, N.; Manshian, B.; Jenkins, G.J.S. NanoGenotoxicology: The DNA damaging potential of engineered nanomaterials. Biomaterials 2009, 30, 3891–3914. [Google Scholar] [CrossRef]
  34. Slobodskova, V.V.; Kukla, S.P.; Chelomin, V.P. An analysis of the quality of the marine environment based on determina-tion of the genotoxicity of DNA in the gill cells of the Yesso Scallop Mizuhopecten yessoensis (Jay, 1856). Russ. J. Mar. Biol. 2015, 41, 495–498. [Google Scholar] [CrossRef]
  35. Hahladakis, J.N.; Velis, C.A.; Weber, R.; Iacovidou, E.; Purnell, P. An overview of chemical additives present in plastics: Migration, release, fate and environmental impact during their use, disposal and recycling. J. Hazard. Mater. 2018, 344, 179–199. [Google Scholar] [CrossRef]
  36. Fauser, P.; Vorkamp, K.; Strand, J. Residual additives in marine microplastics and their risk assessment—A critical review. Mar. Pollut. Bull. 2022, 177, 113467. [Google Scholar] [CrossRef]
  37. Araujo, P.H.H.; Sayer, C.; Giudici, R.; Poço, J.G.R. Techniques for reducing residual monomer content in polymers: A review. Polymer Eng. Sci. 2002, 42, 1442–1468. [Google Scholar] [CrossRef]
  38. Capolupo, M.; Sørensen, L.; Jayasena, K.D.R.; Booth, A.M.; Fabbri, E. Chemical composition and ecotoxicity of plastic and car tire rubber leachates to aquatic organisms. Water Res. 2020, 169, 115270. [Google Scholar] [CrossRef]
  39. Suhrhoff, T.J.; Scholz-Böttcher, B.M. Qualitative impact of salinity, UV radiation and turbulence on leaching of organic plastic additives from four common plastics—A lab experiment. Mar. Pollut. Bull. 2016, 102, 84–94. [Google Scholar] [CrossRef]
  40. León, V.M.; García, I.; González, E.; Samper, R.; Fernández-González, V.; Muniategui-Lorenzo, S. Potential transfer of organic pollutants from littoral plastics debris to the marine environment. Environ. Pollut. 2018, 236, 442–453. [Google Scholar] [CrossRef]
  41. Pan, Y.; Long, Y.; Hui, J.; Xiao, W.; Yin, J.; Li, Y.; Liu, D.; Tian, Q.; Chen, L. Microplastics can affect the trophic cascade strength and stability of plankton ecosystems via behavior-mediated indirect interactions. J. Hazard. Mater. 2022, 430, 128415. [Google Scholar] [CrossRef]
  42. Regoli, F.; Giuliani, M.E. Oxidative pathways of chemical toxicity and oxidative stress biomarkers in marine organisms. Mar. Environ. Res. 2014, 93, 106–117. [Google Scholar] [CrossRef] [PubMed]
  43. Avio, C.G.; Gorbi, S.; Milan, M.; Benedetti, M.; Fattorini, D.; D’Errico, G.; Pauletto, M.; Bargelloni, L.; Regoli, F. Pollutants bioavailability and toxicological risk from microplastics to marine mussels. Environ. Pollut. 2015, 198, 211–222. [Google Scholar] [CrossRef] [PubMed]
  44. Katsumiti, A.; Nicolussi, G.; Bilbao, D.; Prieto, A.; Etxebarria, N.; Cajaraville, M.P. In vitro toxicity testing in hemocytes of the marine mussel Mytilus galloprovincialis (L.) to uncover mechanisms of action of the water accommodated fraction (WAF) of a naphthenic North Sea crude oil without and with dispersant. Sci. Total Environ. 2019, 670, 1084–1094. [Google Scholar] [CrossRef]
  45. Moore, M.N. Do nanoparticles present ecotoxicological risks for the health of the aquatic environment? Environ. Int. 2006, 32, 967–976. [Google Scholar] [CrossRef]
  46. Marigómez, I.; Soto, M.; Cancio, I.; Orbea, A.; Garmendia, L.; Cajaraville, M.P. Cell and tissue biomarkers in mussel, and histopathology in hake and anchovy from Bay of Biscay after the Prestige oil spill (Monitoring Campaign 2003). Mar. Pollut. Bull. 2006, 53, 287–304. [Google Scholar] [CrossRef] [PubMed]
  47. Viarengo, A.; Dondero, F.; Pampanin, D.M.; Fabbri, R.; Poggi, E.; Malizia, M.; Bolognesi, C.; Perrone, E.; Gollo, E.; Cossa, G.P. A biomonitoring study assessing the residual biological effects of pollution caused by the Haven wreck on marine organisms in the Ligurian Sea (Italy). Arch. Environ. Contam. Toxicol. 2007, 53, 607–616. [Google Scholar] [CrossRef]
  48. Neuzil, J.; Zhao, M.; Ostermann, G.; Sticha, M.; Gellert, N.; Weber, C.; Eaton, J.W.; Brunk, U.T. Alpha-tocopheryl succinate, an agent with in vivo anti-tumour activity, induces apoptosis by causing lysosomal instability. Biochem. J. 2002, 362, 709–715. [Google Scholar] [CrossRef]
  49. Kurz, T.; Terman, A.; Gustafsson, B.; Brunk, U.T. Lysosomes in iron metabolism, ageing and apoptosis. Histochem. Cell. Biol. 2008, 129, 389–406. [Google Scholar] [CrossRef] [Green Version]
  50. Mertens, B.; Simon, C.; Van Bossuyt, M.; Onghena, M.; Vandermarken, T.; Van Langenhove, K.; Demaegdt, H.; Van Hoeck, E.; Van Loco, J.; Vandermeiren, K.; et al. Investigation of the genotoxicity of substances migrating from polycarbonate replacement baby bottles to identify chemicals of high concern. Food. Chem. Toxicol. 2016, 89, 126–137. [Google Scholar] [CrossRef]
  51. O’Donovan, S.; Mestre, N.C.; Abel, S.; Fonseca, T.G.; Carteny, C.C.; Cormier, B.; Keiter, S.H.; Bebianno, M.J. Ecotoxicological effects of chemical contaminants adsorbed to microplastics in the clam Scrobicularia plana. Front. Mar. Sci. 2018, 5, 143. [Google Scholar] [CrossRef] [Green Version]
  52. Pannetier, P.; Cachot, J.; Clérandeau, C. Toxicity assessment of pollutants sorbed on environmental sample microplastics collected on beaches: Part I-adverse effects on fish cell line. Environ. Pollut. 2019, 248, 1088–1097. [Google Scholar] [CrossRef]
  53. Gunaalan, K.; Fabbri, E.; Capolupo, M. The hidden threat of plastic leachates: A critical review on their impacts on aquatic organisms. Water Res. 2020, 184, 116170. [Google Scholar] [CrossRef]
  54. Lithner, D.; Larsson ADave, G. Environmental and health hazard ranking and assessment of plastic polymers based on chemical composition. Sci. Total Environ. 2011, 409, 3309–3324. [Google Scholar] [CrossRef] [PubMed]
  55. Yuan, Z.; Nag, R.; Cummins, E. Ranking of potential hazards from microplastics polymers in the marine environment. J. Hazard Mater. 2022, 429, 128399. [Google Scholar] [CrossRef]
  56. Nogueira, L.; Garcia, D.; Trevisan, R.; Sanches, A.L.; da Silva Acosta, D.; Dafre, A.L.; Oliveira, T.Y.; de Almeida, E.A. Biochemical responses in mussels Perna perna exposed to diesel B5. Chemosphere 2015, 134, 210–216. [Google Scholar] [CrossRef] [PubMed]
  57. Belcheva, N.N.; Istomina, A.A.; Dovzhenko, N.V.; Lishavskaya, T.; Chelomin, V.P. Using heavy metal content and lipid peroxidation indicators in the tissues of the mussel Crenomytilus grayanus for pollution assessment after marine environmental remediation. Bull. Environ. Contam. Toxicol. 2015, 95, 481–487. [Google Scholar] [CrossRef] [PubMed]
  58. Istomina, A.; Mazur, A.; Chelomin, V.; Kukla, S.; Slobodskova, V.; Zvyagintsev, A.; Kolosova, L.; Zhukovskaya, A.; Fedorets, Y. The different biomarkers in the assessment of the marine environmental quality using the representative species Mytilus trossulus. Water Air Soil Pollut. 2020, 231, 403. [Google Scholar] [CrossRef]
  59. Santos, J.; Barreto, A.; Sousa, É.M.L.; Calisto, V.; Amorim, M.J.B.; Maria, V.L. The role of nanoplastics on the toxicity of the herbicide phenmedipham, using Danio rerio embryos as model organisms. Environ. Pollut. 2022, 303, 119166. [Google Scholar] [CrossRef]
  60. Zakhartsev, M.V.; Naumenko, N.V.; Chelomin, V.P. Non-methylene-interrupted fatty acids in phospholipids of the membranes of the mussel Crenomytilus grayanus. Russ. J. Mar. Biol. 1998, 24, 183–186. [Google Scholar]
  61. Naudí, A.; Jové, M.; Ayala, V.; Portero-Otín, M.; Barja, G.; Pamplona, R. Membrane lipid unsaturation as physiological adaptation to animal longevity. Front. Physiol. 2013, 17, 372. [Google Scholar] [CrossRef] [Green Version]
  62. Shenai-Tirodkar, P.S.; Gauns, M.U.; Mujawar, M.W.A.; Ansari, Z.A. Antioxidant responses in gills and digestive gland of oyster Crassostrea madrasensis (Preston) under lead exposure. Ecotoxicol. Environ. Saf. 2017, 142, 87–94. [Google Scholar] [CrossRef]
  63. Pandi, P.; Madhuvandhi, J.; Priya, K.K.; Thiagarajan, R.; Gopalakrishnan, S.; Elumalai, S.; Thilagam, H. Weathered polyethylene microplastics exposure leads to modulations in glutathione-S-transferase activity in fish. Front. Mar. Sci. 2022, 9, 990351. [Google Scholar] [CrossRef]
  64. Hansen, B.H.; Altin, D.; Øverjordet, I.B.; Jager, T.; Nordtug, T. Acute exposure of water soluble fractions of marine diesel on Arctic Calanus glacialis and boreal Calanus finmarchicus: Effects on survival and biomarker response. Sci. Total Environ. 2013, 449, 276–284. [Google Scholar] [CrossRef]
  65. Cajaraville, M.P.; Bebianno, M.J.; Blasco, J.; Porte, C.; Sarasquete, C.; Viarengo, A. The use of biomarkers to assess the impact of pollution in coastal environments of the Iberian Peninsula: A practical approach. Sci. Total Environ. 2000, 247, 295–311. [Google Scholar] [CrossRef] [PubMed]
Jmse 11 01446 g001 550
Figure 1. Changes in M. trossulus hemocyte lysosome membrane stability (LMS) exposed to PE and PE-WSF-DF fragments (mean ± standard deviation, n = 20), *—difference from control is significant at p < 0.05.
Figure 1. Changes in M. trossulus hemocyte lysosome membrane stability (LMS) exposed to PE and PE-WSF-DF fragments (mean ± standard deviation, n = 20), *—difference from control is significant at p < 0.05.
Jmse 11 01446 g001
Jmse 11 01446 g002 550
Figure 2. Changes in total oxyradical scavenging capacity (TOSC) in M. trossulus digestive glands and gills exposed to PE and PE-WSF-DF fragments (mean ± standard deviation, n = 15); *—difference from control is significant at p < 0.05.
Figure 2. Changes in total oxyradical scavenging capacity (TOSC) in M. trossulus digestive glands and gills exposed to PE and PE-WSF-DF fragments (mean ± standard deviation, n = 15); *—difference from control is significant at p < 0.05.
Jmse 11 01446 g002
Jmse 11 01446 g003 550
Figure 3. Changes in levels of malonaldehyde (MDA) in M. trossulus digestive gland and gills exposed to PE and PE-WSF-DF fragments (mean ± standard deviation, n = 15).
Figure 3. Changes in levels of malonaldehyde (MDA) in M. trossulus digestive gland and gills exposed to PE and PE-WSF-DF fragments (mean ± standard deviation, n = 15).
Jmse 11 01446 g003
Jmse 11 01446 g004 550
Figure 4. Changes in catalase (A) and glutathione-S-transferase (B) levels in M. trossulus digestive glands and gills exposed to PE and PE-WSF-DF fragments (mean ± standard deviation, n = 15); *—difference from control is significant at p < 0.05.
Figure 4. Changes in catalase (A) and glutathione-S-transferase (B) levels in M. trossulus digestive glands and gills exposed to PE and PE-WSF-DF fragments (mean ± standard deviation, n = 15); *—difference from control is significant at p < 0.05.
Jmse 11 01446 g004
Jmse 11 01446 g005 550
Figure 5. Changes in levels of DNA damage in M. trossulus digestive gland and gills exposed to PE and PE-WSF-DF fragments (mean ±standard deviation, n = 15); *—difference from control is significant at p < 0.05.
Figure 5. Changes in levels of DNA damage in M. trossulus digestive gland and gills exposed to PE and PE-WSF-DF fragments (mean ±standard deviation, n = 15); *—difference from control is significant at p < 0.05.
Jmse 11 01446 g005
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Dovzhenko, N.V.; Chelomin, V.P.; Mazur, A.A.; Slobodskova, V.V.; Istomina, A.A.; Kukla, S.P. Biomarker Effects of Diesel Fuel Hydrocarbons Absorbed to PE-Plastic Debris on Mussel Mytilus trossulus. J. Mar. Sci. Eng. 2023, 11, 1446. https://doi.org/10.3390/jmse11071446

AMA Style

Dovzhenko NV, Chelomin VP, Mazur AA, Slobodskova VV, Istomina AA, Kukla SP. Biomarker Effects of Diesel Fuel Hydrocarbons Absorbed to PE-Plastic Debris on Mussel Mytilus trossulus. Journal of Marine Science and Engineering. 2023; 11(7):1446. https://doi.org/10.3390/jmse11071446

Chicago/Turabian Style

Dovzhenko, Nadezda Vladimirovna, Victor Pavlovich Chelomin, Andrey Alexandrovich Mazur, Valentina Vladimirovna Slobodskova, Aleksandra Anatolyevna Istomina, and Sergey Petrovich Kukla. 2023. "Biomarker Effects of Diesel Fuel Hydrocarbons Absorbed to PE-Plastic Debris on Mussel Mytilus trossulus" Journal of Marine Science and Engineering 11, no. 7: 1446. https://doi.org/10.3390/jmse11071446

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop