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Article

Anthropogenic Microparticles in Aquaculture and Wild Fish: A Case Study of Three Commercially Important Species in the Eastern Mediterranean

by
Aikaterini Kostoula
1,
Eugenia Moschou-Kounopioti
2,
Niki Milatou
1,* and
Persefoni Megalofonou
1,2
1
Department of Biology, National and Kapodistrian University of Athens, Panepistimiopolis, Ilissia, 15784 Athens, Greece
2
Interinstitutional Postgraduate Program NKUA-HCMR Oceanography and Management of Marine Environment, National and Kapodistrian University of Athens, Panepistimiopolis, Ilissia, 15784 Athens, Greece
*
Author to whom correspondence should be addressed.
Fishes 2025, 10(10), 492; https://doi.org/10.3390/fishes10100492
Submission received: 1 September 2025 / Revised: 14 September 2025 / Accepted: 23 September 2025 / Published: 2 October 2025
(This article belongs to the Section Biology and Ecology)
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Abstract

Over the past decade, increasing attention has been given to the impacts of anthropogenic microparticle (AM) pollution on marine ecosystems. This study investigates AM ingestion in three commercially important fish species—Sparus aurata Linnaeus, 1758, Dicentrarchus labrax Linnaeus, 1758, and Boops boops Linnaeus, 1758—collected from both wild and farmed populations in Greek marine and lagoon environments. A total of 60 specimens were sampled from the Messolonghi Lagoon, Rhodes Island, and the Cyclades. AM were detected in 61.7% of the individuals analyzed. The mean number of ingested items per individual was 1.1 ± 1.2 in B. boops, 1.0 ± 1.7 in wild and 2.3 ± 2.1 in farmed S. aurata, and 2.5 ± 3.1 in wild and 3.6 ± 2.2 in farmed D. labrax. Ingestion ranged from 0 to 9 items per fish. No significant correlations were found between fish size and either the number or the size of ingested AM in any species. The ingested AM were primarily classified as fibers and fragments, displaying variability in size and color. Black was the dominant color across all species, followed by red and blue, while yellow was rarely observed. A statistically significant difference in the mean size of AM was recorded between wild and farmed D. labrax, whereas no such difference was observed for S. aurata. Overall, these findings provide new evidence on AM contamination in seafood species and highlight their occurrence in both natural and aquaculture environments of the eastern Mediterranean.
Keywords:
anthropogenic microparticle ingestion; Sparus aurata; Dicentrarchus labrax; Boops boops; aquaculture; eastern Mediterranean sea
Key Contribution: This study provides the first comparative investigation of anthropogenic microparticle (AM) ingestion in three commercially important fish species (S. aurata, D. labrax, and B. boops) from both wild and farmed populations in Greek marine and lagoon environments. AM were present in 61.7% of individuals, with fibers dominating and black being the most frequent color. While ingestion rates did not differ significantly between wild and aquaculture-origin fish, farmed individuals—especially D. labrax—tended to ingest particles of larger size. These findings underscore the ubiquity of AM contamination across natural and aquaculture settings in the eastern Mediterranean and highlight potential implications for seafood safety and human exposure.

1. Introduction

The presence of marine litter in the oceans is an increasingly critical threat to the health of marine ecosystems [1]. The majority of this litter consists of plastic items, which are pervasive due to their durability, low cost, and widespread use [2]. Plastics are synthetic polymeric compounds, primarily derived from petrochemical sources, and are characterized by high molecular weight and plasticity. Various chemical additives are incorporated during manufacturing to enhance the performance and durability of plastic products [3].
For decades, plastics were indiscriminately discarded into the oceans, often regarded as a vast “sink” for waste, without consideration for their long-term ecological consequences [4]. These plastics originate from the degradation of clothing, furniture, and sanitary products, and are introduced into the marine environment primarily through wastewater and runoff from rivers and lakes. Their accumulation is particularly pronounced in coastal areas subject to intense human activity [4].
Globally, approximately 350–400 million tonnes of plastic are produced annually, with an estimated 19–23 million tonnes entering aquatic ecosystems each year [5,6]. The Mediterranean Sea, with a surface area of about 2.5 million km2, is considered a major accumulation zone of floating plastic debris and one of the most impacted semi-enclosed seas worldwide [4,7].
Anthropogenic microparticle (AM) debris in the marine environment often breaks down into very small particles known as microplastics (MP). The most widely accepted definition, as established by NOAA, categorizes microplastics as plastic particles less than 5 mm in their largest dimension. Due to their small size, AM can be ingested by a wide range of marine organisms, raising concerns about their ecological and physiological effects [4,8].
Even smaller particles, known as nanoplastics, range in size from 0.001 µm to 0.1 µm. These are considered potentially more hazardous than AM because they are small enough to cross biological membranes and accumulate within cells [9].
The impact of marine debris ingestion is particularly pronounced in areas influenced by oceanic convergence currents, where floating litter tends to accumulate [10]. Although this phenomenon occurs globally, its extent is more pronounced in certain oceanic regions [2,11,12]. In the Mediterranean Sea, AM debris has been detected in the gastrointestinal tracts of a wide range of marine taxa, including sea turtles, teleost fishes, elasmobranchs, cetaceans, and invertebrates [13,14,15]. Several commercially important fish species have also been investigated for AM ingestion, such as B. boops, Mullus barbatus Linnaeus, 1758, Pagellus erythrinus Linnaeus, 1758, Sardina pilchardus Walbaum, 1792, and S. aurata, with reported frequencies of occurrence ranging from 30% to 80% depending on the region [16].
Wang and Liu [4] identified four main types of adverse effects of AM ingestion on marine organisms. First, ingestion can lead to reduced mobility, impaired reproduction, gastrointestinal obstruction, and malnutrition due to blockage of the esophagus. Second, ingested AM can leach hazardous chemicals that accumulate in tissues, potentially causing toxicological effects. Third, entanglement in plastic debris remains a serious threat to marine fauna, often resulting in injury or death, especially in birds and large vertebrates. Finally, with the global expansion of fisheries and aquaculture, plastic equipment (e.g., nets, ropes, buoys) contributes further to AM pollution in the marine environment.
The present study provides a comparative assessment of AM ingestion in three commercially important fish species (S. aurata, D. labrax, and B. boops) from Greek marine and lagoon ecosystems, by analyzing both wild and farmed individuals. The innovative element of this research lies in the direct, side–by–side comparison between wild and aquaculture-origin fish of the same species, combined with a detailed characterization of AM types, colors, and sizes. While AM ingestion has previously been reported in these species in the Mediterranean, this is the first study focusing on a comparative approach of wild and farmed populations from different Greek habitats. Specifically, it aims to: (i) assess the occurrence and frequency of AM in both wild and farmed individuals; (ii) characterize the types, colors, and sizes of AM ingested; and (iii) explore potential differences between wild and aquaculture-origin fish, with particular attention to the role of aquaculture-related AM sources.

2. Materials and Methods

2.1. Study Area

A total of 60 fish specimens were collected between February and December 2019 from three regions in Greece (Messolonghi Lagoon, Cyclades, and Rhodes Island), located in the eastern Mediterranean Sea. Specifically, 10 farmed S. aurata and 10 farmed D. labrax were obtained from a commercial fish farm in Rhodes; 10 wild S. aurata and 10 wild D. labrax were collected from the Messolonghi Lagoon; and 20 wild B. boops specimens were sampled in the Cyclades region (Figure 1).
All fish were processed at the time of slaughter in accordance with European Union animal welfare regulations (EC Regulation No. 1099/2009 on the Protection of Animals at the Time of Killing). Following collection, all specimens were stored at −20 °C until further analysis.

2.2. Extraction of Anthropogenic Microparticles (AM) from Fish Tissues

Prior to dissection, fish specimens were defrosted at room temperature. For each specimen, total length (TL) and round weight (RW, i.e., total body weight) were recorded (Table 1). Subsequently, the stomach and intestine were dissected, and their contents were removed and weighed (wet weight).
Gastrointestinal contents were initially inspected visually under a stereomicroscope to detect any suspected AM. To remove organic matter and facilitate the identification of AM, samples were digested using 30% hydrogen peroxide (H2O2), following the protocol established by the MEDSEALITTER project (https://publications.jrc.ec.europa.eu/repository/handle/JRC83985, accessed on 23 September 2025).
Specifically, the digestive tract content of each specimen was transferred into a clean glass beaker and mixed with 30% H2O2 (ChemLab, Ghent, Belgium) at a ratio of 1:20 (w/v). The mixture was heated on a hot plate at 60 °C until the peroxide was fully evaporated and the organic material was completely digested.
After digestion, each sample was diluted with 100 mL of Milli-Q purified water (Merck Millipore, Darmstadt, Germany), gently stirred, and vacuum-filtered through glass fiber filters (Whatman GF/C, 1.2 µm pore size, 47 mm diameter; Cytiva, Marlborough, MA, USA). Upon completion of filtration, filters were carefully transferred into clean Petri dishes and left to dry at room temperature until further analysis.

2.3. Anthropogenic Microparticle Detection and Quantification

In this study, AM were identified based on visual characteristics (shape, size, color) under a stereomicroscope, without spectroscopic confirmation of polymer type.
After drying, all filters were examined under a Nikon SMZ-2T binocular stereomicroscope (Nikon Instruments Inc., Melville, NY, USA) equipped with a digital camera (INFINITYlite, Teledyne Lumenera, Ottawa, ON, Canada). AM were photographed, counted, and measured using the Image Analysis Pro Plus 6.0 software.
For each particle, length, width, color, and type (e.g., fragment, fiber) were recorded. Identified AM were further categorized into three size classes: <0.05 mm, 0.05–1.0 mm, and 1.0 mm (https://publications.jrc.ec.europa.eu/repository/handle/JRC83985, accessed on 23 September 2025).
AM abundance was quantified in three ways: (a) as the average number of AM per individual, considering the total number of individuals analyzed, (b) as the average number of items per individual, calculated only for those specimens that contained AM, and (c) as the average number of items per gram of wet weight of gastrointestinal content, for individuals containing AM (https://publications.jrc.ec.europa.eu/repository/handle/JRC83985, accessed on 23 September 2025).

2.4. Precautions and Quality Control

To minimize the risk of contamination, all glassware and equipment were thoroughly rinsed with purified (Milli-Q) water prior to use. During the initial observation of gastrointestinal contents, each Petri dish was covered with a glass lid to prevent airborne contamination. Throughout the digestion process, as well as during any periods when samples were not actively being processed, they were kept covered with aluminum foil (https://publications.jrc.ec.europa.eu/repository/handle/JRC83985, accessed on 23 September 2025).
All filters were protected with glass covers during microscopic examination. Sample handling, including rinsing and vacuum filtration, was carried out inside a fume cupboard to further reduce exposure to airborne particles. Procedural blanks were included at every stage of the analysis. Any particles that matched those observed in the blanks were excluded from the dataset and considered potential laboratory contamination (https://publications.jrc.ec.europa.eu/repository/handle/JRC83985, accessed on 23 September 2025).

2.5. Statistical Analysis

Descriptive statistics (mean, standard deviation, and range) were calculated for all measured parameters. Prior to further analysis, all datasets were tested for normality and homogeneity of variances. For normally distributed variables, Student’s t-test was applied to assess differences in mean values between groups. When normality assumptions were not met, non-parametric tests were used: the Kolmogorov–Smirnov or Mann–Whitney U test for comparisons between two groups, and the Kruskal–Wallis test for comparisons among more than two groups.
Relationships between variables (e.g., fish length and number or size of AM) were examined using regression analysis. A significance level of p ≤ 0.05 was adopted for all statistical tests.
All statistical analyses and graphical outputs were performed using Statgraphics Centurion XVII and Microsoft Excel (Microsoft Corporation, Redmond, WA, USA).

3. Results

3.1. Anthropogenic Microparticle Ingestion

In total, 115 AM were detected in the gastrointestinal tracts of 11 B. boops (55%), 6 wild S. aurata (60%), 8 farmed S. aurata (80%), 4 wild D. labrax (40%), and 8 farmed D. labrax (80%) (Table 1).
The average number of AM per individual, considering all examined fish, was 1.1 ± 1.2 in B. boops, 2.5 ± 3.1 in wild D. labrax, 3.6 ± 2.2 in farmed D. labrax, 1.0 ± 1.7 in wild S. aurata, and 2.3 ± 2.1 in farmed S. aurata. No statistically significant differences were found among species or between wild and farmed groups (t-test, p > 0.05).
When considering only those individuals in which AM were present, the average number of particles per individual was 1.9 ± 1.0 in B. boops, 4.2 ± 2.9 in wild D. labrax, 4.5 ± 1.3 in farmed D. labrax, 2.5 ± 1.9 in wild S. aurata, and 2.9 ± 1.9 in farmed S. aurata. Again, no significant differences were observed among species or between wild and farmed individuals (t-test, p > 0.05), except for a statistically significant difference between wild D. labrax and B. boops (t-test, p < 0.05).
The average number of AM per gram of gastrointestinal content (wet weight), considering only individuals with AM, was 11.4 ± 6.7 items/g in B. boops, 21.5 ± 17.6 in wild D. labrax, 20.0 ± 12.1 in farmed D. labrax, 14.3 ± 13.5 in wild S. aurata, and 7.7 ± 5.2 in farmed S. aurata. No statistically significant differences were detected among species or between wild and farmed individuals (t-test, p > 0.05), with the exception of a significant difference between farmed D. labrax and farmed S. aurata (Mann–Whitney test, p < 0.05).
Furthermore, no significant differences were found between wild and farmed groups of D. labrax or S. aurata in terms of AM abundance (t-test, p > 0.05). The highest occurrence rates were observed in farmed D. labrax and farmed S. aurata (80% each), while wild D. labrax showed the lowest frequency (40%). Overall, farmed fish tended to exhibit a higher number of ingested AM per individual than their wild counterparts, though without statistical significance. No correlation was observed between the number of ingested AM and fish size (length or weight) within any species (Spearman’s correlation, p > 0.05).

3.2. Anthropogenic Microparticle Characterization (Shape, Size, and Color)

AM retrieved from the gastrointestinal contents of the examined specimens exhibited variation in shape, size, and color. Fibers were the dominant shape category across all fish species, in both wild and farmed individuals (Figure S1). Among wild specimens, the proportion of fibers and fragments was 86% and 14% in B. boops, and 100% fibers in both D. labrax and S. aurata. In farmed specimens, fibers accounted for 97% of particles in D. labrax and 87% in S. aurata, with fragments comprising the remaining 3% and 13%, respectively (Figure 2a and Figure 3a).
Regarding particle size, AM < 0.05 mm were the most abundant size class in wild individuals, comprising 50% of MP in S. aurata, 60% in D. labrax, and 62% in B. boops. In contrast, farmed fish were more frequently associated with AM in the 0.05–1.0 mm range, which represented 67% of particles in D. labrax and 78% in S. aurata (Figure 2b and Figure 3b). A statistically significant difference in AM size distribution was observed between wild and farmed D. labrax (Kolmogorov–Smirnov test, p < 0.05), whereas no such difference was found for S. aurata (p > 0.05). Furthermore, no significant differences in AM size were detected among the three species overall (Kruskal–Wallis test, p > 0.05). No correlation was found between AM size and fish total length in any species (Spearman’s r, p > 0.05).
In terms of color, black AM were the most dominant across all species, representing 43.5% of the total particles recorded. The next most frequent colors were red (24.3%) and blue (19.1%), while yellow particles were rare, comprising only 1% of the total (Figure 2c and Figure 3c).
In total, 106 out of 115 recorded AM (92.2%) were classified as fibers, confirming their overwhelming predominance across all fish categories. Based on visual quantification, the most common size class overall was 0.05–1.0 mm, comprising approximately 52% of all particles, followed by particles < 0.05 mm (about 45%). In terms of color distribution, of the 115 particles recorded, 50 were black, 28 red, 22 blue, and only one yellow, while the remaining 14 belonged to other color categories.

4. Discussion

4.1. Anthropogenic Microparticle Ingestion Across Species and Habitats

AM were detected in all fish species examined in this study (B. boops, D. labrax, and S. aurata), in both wild and farmed individuals. Particles varied in shape, size, and color, and were found within the stomach and gastrointestinal tract of the specimens. These findings confirm the widespread presence of AM pollution in all sampling areas investigated. Humans are also exposed to AM through seafood consumption, with recent estimates suggesting ingestion of up to 121,000 particles per year per adult [17].
The frequency of occurrence of ingested AM ranged from 40% to 80%, consistent with the literature [16,17,18]. AM abundance per individual, across all examined individuals, was comparable among species, with wild specimens showing values between 1.1 ± 1.2 and 2.5 ± 3.1 items per individual and farmed specimens between 2.3 ± 2.1 and 3.6 ± 2.2. The highest frequency of occurrence (80%) was observed in farmed D. labrax and S. aurata.
B. boops presented a percentage of AM ingestion (55%), which is higher than the 46% reported by Garcia-Garin et al. [17] in the NW Mediterranean and 47% by Tsangaris et al. [19] in the Eastern Mediterranean. Moreover, Nadal et al. [20] reported even higher ingestion rates (68%) for this species in the Balearic Islands and 56% by Sbrana et al. [21] in the Tyrrhenian Sea. Comparable results have also been obtained in other areas of the western Mediterranean, where 16–37% of the individuals examined contained AM [22].
Similar AM ingestion levels in B. boops have been reported by Garcia-Garin et al. [17] from the Catalan coast (1.68 ± 0.31 items/individual) and by Sbrana et al. [21] from the Tyrrhenian Sea (1.80 ± 0.20 items/individual). In contrast, Tsangaris et al. [19] found significantly lower values in B. boops from the Ionian Sea (0.23 ± 0.05) and the Saronikos Gulf (0.43 ± 0.13) (Table 2). In the Cabrera MPA, however, Compa et al. [23] reported much higher values in B. boops, with an average of 14.5 ± 7.8 items/individual. Farmed S. aurata and D. labrax from Spain have also been reported to ingest high AM loads, ranging between 2.0 ± 0.3 and 5.4 ± 4.2 items per individual [24,25]. However, Savoca et al. [26] reported a much lower abundance of microparticles in farmed S. aurata (0.21 items/individual in fry and 1.3 items/individual in adults).
Considering only individuals containing AM, the average number of AM per individual was relatively similar between B. boops (1.9 ± 1.0) and wild S. aurata (2.5 ± 1.9), and between wild and farmed S. aurata (2.5 ± 1.9 vs. 2.9 ± 1.9). A similar trend was observed for D. labrax (wild: 4.2 ± 2.9; farmed: 4.5 ± 1.3). However, a statistically significant difference was found between wild D. labrax and B. boops. These findings are generally consistent with those reported in previous studies [17,24,25]. Conversely, Anastasopoulou et al. [16] documented substantially higher values (7.3 ± 6.6) in wild S. aurata from the Adriatic Sea, likely reflecting the high AM load of that region (Table 2). In line with these observations, El-Sayed et al. [30] reported that the average MPs concentration was significantly higher in S. aurata (38.3 ± 28.4 items/fish) than in B. boops (20.0 ± 20.9 items/ fish). In contrast to the high density of plastics in S. aurata, this species exhibited the lowest composition of plastics in their digestive systems. B. boops had the highest plastic contents (43.0%), followed by S. rivulatus, D. vulgaris, and S. scriba (25.3, 18.2, and 16.2%, respectively). This pattern was attributed to the smaller size of fish, which reduces their ability to select appropriate food, thereby influencing stomach contents.
These results are broadly consistent with recent literature. For instance, Mosconi et al. [28] investigated seabream and seabass from offshore aquaculture systems in the Mediterranean and reported AM ingestion in 38% of individuals, with an average of 0.51 ± 0.78 anthropogenic particles/fish and a maximum of 3 particles per individual. Although ingestion occurred mainly in the gastrointestinal tract, a small number of anthropogenic particles were also found in liver and muscle tissue, raising concerns regarding human dietary exposure even from aquaculture products (Table 2). In addition, Matias et al. [29] demonstrated significantly elevated AM burdens in RAS-reared fish, suggesting that farming method and system design substantially influence AM ingestion risk.
No significant differences in AM ingestion were observed between wild and farmed specimens of D. labrax and S. aurata, suggesting that neither the origin (wild vs. aquaculture) nor the specific sampling site had a major influence on AM intake. These results align with findings by Digka et al. [27], who reported no differences in AM contamination between wild and farmed mussels from the northern Ionian Sea. The observed similarities may be attributed to multiple factors: (i) aquaculture facilities are typically located in coastal areas already exposed to high AM loads, subjecting farmed fish to conditions similar to those of wild populations; (ii) feeding habits and diets partly overlap between wild and aquaculture fish, potentially leading to comparable ingestion rates; and (iii) hydrodynamic conditions and water exchange between lagoon and marine environments may promote a broadly uniform distribution of anthropogenic particles. Collectively, these factors provide a plausible explanation for the absence of significant differences across habitats and fish origins.
Although the studied species differ in their trophic levels and feeding strategies (https://www.fishbase.se, accessed on 1 June 2025), these differences did not translate into significant variation in AM ingestion, suggesting that environmental exposure may be the dominant driver. In particular, B. boops is a planktivorous species with a flexible diet and high susceptibility to visual confusion between prey and anthropogenic particles. Several Mediterranean studies have reported high AM ingestion frequencies in this species [17,19,20,31], with marked spatial variability linked to local environmental conditions. This ecological profile helps to explain both the elevated ingestion rates often recorded for B. boops and the variability observed across regions.
In our study, the overall distribution of AM was broadly similar between wild and farmed specimens (Figure 2 and Figure 3). This suggests that both natural habitats and aquaculture facilities expose fish to comparable types of anthropogenic particles, in line with observations from the Mediterranean reported by Digka et al. [27].
It is important to highlight that comparisons across studies are often challenging due to methodological differences in AM extraction, identification, and reporting. Discrepancies in observed AM abundance, even for the same species, may be attributed to actual variations in environmental AM contamination, but also to inconsistent protocols among research groups. Consequently, there is a pressing need to standardize methodologies in AM research to improve the reliability of assessments and ensure comparability across studies and regions. Moreover, oceanographic processes (e.g., coastal currents, riverine inputs, and benthic resuspension) strongly influence AM concentrations and size distribution in the water column and sediments. Such processes can generate marked spatial heterogeneity even between nearby sites, potentially affecting the ingestion levels observed in the present study. Therefore, direct comparisons among sampling sites should be interpreted with caution.

4.2. Anthropogenic Microparticle Characterization

The shape, color, and size of the AM identified in the fish specimens analyzed in this study are generally consistent with findings from previous research in the Mediterranean Sea [17,18,19,21,24,25,27,28,29]. The concentration and composition of AM in the marine environment are known to vary depending on local conditions, sources, and hydrodynamics.
In our study, fibers represented the dominant shape category in all species and across both wild and farmed groups, ranging from 60% to 100% of the total anthropogenic particles, while fragments accounted for 0% to 40%. These results align with the majority of studies on AM ingestion in fish, which also report a higher prevalence of fibers (Table 2) [2,32,33,34,35,36,37]. The authors attributed this to the prevalence of synthetic textiles and fishing/aquaculture gear as likely sources of contamination, especially in offshore cages. In addition, the dominance of fibers across sites suggested a widespread exposure to textile-related microdebris, while spatial variability underscored the influence of localized environmental conditions [28]. Similarly, Mosconi et al. [28] found that fibers constituted 68% of all ingested anthropogenic particles in farmed seabream and seabass, attributing this to sources such as synthetic textiles and aquaculture gear. However, no direct comparison between pelagic and benthic species was provided.
In farmed S. aurata and D. labrax from the Canary Islands, Sánchez-Almeida et al. [25] observed a predominance of microfibers (>97%). The authors suggested that such patterns reflected homogeneous exposure in fish farms and potentially linked the sources to wastewater discharge points widely distributed around the islands.
In agreement with our findings, fragments and fibers were also the most abundant AM shapes reported in B. boops from other Mediterranean areas, such as the northern Catalan coasts and South Sardinia [19] and the southern Tyrrhenian Sea [31]. Regarding color, Bottari et al. [31] observed a predominance of transparent, black, and white particles in bogues, which they attributed to a potential resemblance to the exoskeletons of crustaceans or gelatinous plankton. Such results, together with the geographical variability in AM color reported by Tsangaris et al. [19], highlight both ecological feeding interactions and differences in local contamination sources across the Mediterranean.
Similar results were reported by Rios-Fuster et al. [22] for B. boops in the western Mediterranean, where fibers were also the most abundant type, and transparent particles were predominant, followed by blue. In their study, the size of the ingested particles ranged from 0.51 to 13.11 mm, confirming that B. boops mainly ingests fibrous microparticles. Moreover, in the Cabrera MPA, Compa et al. [23] reported a predominance of fibers (55.2%), followed by fragments (34.5%) and films (10.3%).
However, contrasting results have been reported in some studies, where fragments exceeded fibers, particularly in both fish and mussel species [29,38,39,40,41]. Such discrepancies may reflect variations in dominant AM sources and regional waste management practices [40].
With regard to color, black was the most frequent among all species examined, except in farmed S. aurata, where blue AM slightly predominated. Several studies report that blue is often the most common color of AM ingested by fish [17,18,25,27,29] with Reinold et al. [18] suggesting that blue items may be mistaken for prey due to their visual resemblance to water. In farmed D. labrax and S. aurata from the Canary Islands, Sánchez-Almeida et al. [25] also found colourless and blue fibers as the predominant categories, with a size distribution peaking around 0.8–1.2 mm. According to Hacısalihoğlu [42], the most color variation was observed in D. labrax. It was determined that blue, purple, and transparent colors were dominant in this species, while red was not observed. In D. labrax, the most common MP types were line and bead (30%), followed by fiber (20%), and fragment and film (10%) respectively. Savoca et al. [43] also revealed the occurrence of man-made cellulose fibers in B. boops from the Northern Sicilian coasts. Of the 30 specimens examined, 63.3% had ingested fiber items, ranging from 1 to 10 per specimen, with an average of 2.7 items/individual. Fiber length ranged from 0.5 to 30 mm, with the vast majority being black (95%) and a small proportion red (5%). These results corroborate our observations of limited variability in AM shape and color between farmed and wild fish and further support the role of aquaculture facilities as sinks of textile-derived microfibers. On the other hand, the near absence of yellow particles in our study may be attributed to the fact that yellow often serves as a warning color in nature and may therefore be avoided by fish [18]. This pattern is mirrored in Mosconi et al. [28], who found that black particles accounted for 41% of all ingested AM, followed by transparent and red. These colors are typically linked to degraded synthetic ropes, nets, or urban textile fibers. Similarly, Savoca et al. [26] observed that in farmed S. aurata, microfibers represented the dominant type of ingested particles. In particular, black (46.15%) and azure (20.5%) were the most frequently recorded colors, followed by lower proportions of other colors. These findings further support the evidence that aquaculture fish are exposed to AM, with microfibers being the most common form. In their study, particle sizes ranged from 0.24 to 8.86 mm, which is comparable to the size range recorded in the present work. In addition, their reported size range overlaps with ours, supporting the comparability of measurements.
As for AM size, the smallest size class (<0.05 mm) was the most prevalent in wild specimens, followed by particles measuring 0.05–1.0 mm. Farmed specimens, particularly D. labrax, exhibited a higher average AM size than wild individuals. This difference was statistically significant only in D. labrax, while no significant difference was found in S. aurata. Nonetheless, due to the relatively small sample size of the present study, these results should be interpreted with caution. Larger-scale investigations are required to validate potential size-related trends between wild and aquaculture-origin fish.
Similar patterns have been reported in the literature. Mosconi et al. [28] recorded anthropogenic particles ranging from 0.12 mm to over 3.0 mm in farmed seabream and seabass, with Greek aquaculture fish exhibiting a predominance of anthropogenic particles (>5 mm), whereas smaller particles were more common in samples from Turkey. This variation was attributed to differences in aquaculture practices, feed handling, and environmental exposure.
In parallel, Digka et al. [27] emphasized the role of habitat and hydrodynamics in shaping particle size ingestion. Their results showed that fish from lagoon environments ingested a higher proportion of larger, sediment-associated particles, while pelagic species exposed to surface waters primarily ingested smaller, suspended AM. Such findings support the hypothesis that environmental conditions and feeding ecology may influence not only the presence but also the size spectrum of ingested AM.

4.3. Human Exposure and Food Safety Implications

Although our study focused on gastrointestinal contents, the potential for translocation of AM to edible tissues is of increasing concern. Mosconi et al. [28] detected anthropogenic particles in fish muscle at a low frequency but still warned of potential risks due to chronic exposure. These findings raise food safety questions, particularly regarding species consumed whole (e.g., small fish) or where edible tissues may accumulate plastic-associated chemicals such as phthalates or persistent organic pollutants (POPs). The bioavailability and toxicological relevance of these compounds require further research, as highlighted by Mosconi et al. [28].
Considering the recommendations of the EFSA, EUMOFA and NOAA for the consumption of fish by different age groups, the intake of AM—calculated by ingestion of three commonly consumed fish species (D. labrax, Trachurus spp., and Scomber colias Gmelin, 1789)—ranges from 112 to 842 particles per year according to EFSA and from 518 to 3078 particles per year per capita according to EUMOFA and NOA (https://www.efsa.europa.eu/en; https://www.ncei.noaa.gov/, accessed on 1 June 2025).
Similarly, Gao et al. [44] documented the presence of anthropogenic particles in the muscle of commercial fish species from Malaysia, with an estimated annual dietary intake of 1062 items per capita through fish consumption. The majority of these particles were blue fibers smaller than 0.5 mm, further highlighting the potential health risks associated with seafood consumption.

4.4. Methodological Considerations and Future Directions

Both our study and recent literature stress the need for methodological harmonization. Differences in digestion protocols, filter pore size, polymer identification, and classification criteria lead to variability in reported AM levels, even within similar environments or species. Without standardization, comparison across studies remains challenging. One limitation of the present study is the absence of chemical analysis of the recovered AM (e.g., µFTIR or Raman spectroscopy). While the visual classification applied here provides reliable estimates of AM abundance and morphology, spectroscopic confirmation would allow for precise polymer identification and assessment of associated additives. Future research should: (a) expand sample sizes and sampling locations to include seasonal and depth-related variability, (b) examine edible tissues systematically to assess real exposure levels in consumers, (c) use spectroscopic techniques (e.g., µFTIR, Raman) to identify polymers and associated additives more precisely, and (d) integrate environmental monitoring (e.g., sediment, water column) to link AM availability with ingestion patterns. In addition, future studies should integrate chemical characterization to better understand the sources, composition, and potential toxicological risks of ingested AM.

5. Conclusions

This study provides robust evidence of the widespread occurrence of AM in three commercially important fish species from the Eastern Mediterranean, including both wild and farmed individuals. AM were detected in all species examined (B. boops, D. labrax, and S. aurata), with frequencies of occurrence ranging from 40% to 80%. No significant differences in ingestion were found between wild and farmed individuals of D. labrax and S. aurata, indicating that origin (wild vs. aquaculture) may not be a determining factor of exposure.
Fibers were the dominant type of ingested particles, and black was the most frequent colour. Wild fish tended to ingest smaller AM, whereas farmed individuals—particularly D. labrax—showed a tendency toward larger particles. These findings underscore the complexity of AM pollution and the role of both environmental exposure and aquaculture conditions.
Importantly, the presence of AM even in farmed seafood highlights potential concerns for food safety and human exposure, reinforcing the need for continued monitoring. At the same time, the variability among studies demonstrates the urgent need for methodological harmonization in sampling, extraction, and quantification protocols, to enable reliable comparisons across regions and species.
Future research should focus on Mediterranean wild and farmed species—particularly S. aurata, D. labrax, and B. boops—through expanded sample sizes, seasonal and spatial coverage, and investigations of potential physiological and toxicological impacts. Such efforts will be critical for clarifying the risks posed by AM contamination to both marine ecosystems and human consumers.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/fishes10100492/s1, Figure S1: Stereomicroscope photograph of a black microfiber found in the gastrointestinal tract of Dicentrarchus labrax analyzed in this study.

Author Contributions

A.K.: Methodology, investigation, data curation. E.M.-K.: Methodology, investigation. N.M.: Writing—original draft preparation, conceptualization, visualization, methodology, formal analysis, data curation, writing—reviewing and editing. P.M.: Conceptualization, methodology, supervision, resources, writing—reviewing and editing. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Ethical review and approval were waived for this study because it did not involve any experiments on animals.

Data Availability Statement

The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding author(s). Web pages: https://medsealitter.interreg-med.eu, accessed on 1 June 2025; https://www.efsa.europa.eu/en, accessed on 1 June 2025; https://www.ncei.noaa.gov/, accessed on 1 June 2025; https://www.fishbase.se, accessed on 1 June 2025.

Acknowledgments

The authors wish to express their gratitude to A. Maranti and C. Grammatikopoulos for their valuable assistance with field sampling and sample processing. Special thanks are also extended to the commercial fishermen and aquaculture producers for their cooperation and support during the sampling campaigns.

Conflicts of Interest

The authors declare no conflicts of interest.

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Fishes 10 00492 g001
Figure 1. Map illustrating the sampling locations for anthropogenic microparticle ingestion analysis in three commercial fish species (B. boops, D. labrax, and S. aurata) across Greece, including sites from the Messolonghi lagoon, Rhodes Island, and the Cyclades.
Figure 1. Map illustrating the sampling locations for anthropogenic microparticle ingestion analysis in three commercial fish species (B. boops, D. labrax, and S. aurata) across Greece, including sites from the Messolonghi lagoon, Rhodes Island, and the Cyclades.
Fishes 10 00492 g001
Fishes 10 00492 g002
Figure 2. Percentage distribution of anthropogenic microparticle (a) shape (fibers, fragments), (b) size classes (<0.05 mm, 0.05–1.0 mm, >1.0 mm), and (c) color (black, blue, red, yellow) identified in wild fish specimens (B. boops, D. labrax, and S. aurata) collected from Greek coastal waters.
Figure 2. Percentage distribution of anthropogenic microparticle (a) shape (fibers, fragments), (b) size classes (<0.05 mm, 0.05–1.0 mm, >1.0 mm), and (c) color (black, blue, red, yellow) identified in wild fish specimens (B. boops, D. labrax, and S. aurata) collected from Greek coastal waters.
Fishes 10 00492 g002
Fishes 10 00492 g003
Figure 3. Percentage distribution of anthropogenic microparticle (a) shape, (b) size classes, and (c) color detected in farmed fish specimens (D. labrax and S. aurata) from aquaculture facilities in Greece.
Figure 3. Percentage distribution of anthropogenic microparticle (a) shape, (b) size classes, and (c) color detected in farmed fish specimens (D. labrax and S. aurata) from aquaculture facilities in Greece.
Fishes 10 00492 g003
Table 1. Morphometric characteristics (TL = total length; RW = round weight, i.e., total body weight), frequency (%) of anthropogenic microparticle (AM) occurrence, and AM abundance (mean ± SD) in three commercial fish species (B. boops, D. labrax, and S. aurata) collected from Greece. AM abundance is expressed as: (a) average number of AM per individual among all examined individuals, (b) average number of AM per individual among individuals containing AM, and (c) average number of AM per gram wet weight (w.w.) of gastrointestinal content among individuals containing AM.
Table 1. Morphometric characteristics (TL = total length; RW = round weight, i.e., total body weight), frequency (%) of anthropogenic microparticle (AM) occurrence, and AM abundance (mean ± SD) in three commercial fish species (B. boops, D. labrax, and S. aurata) collected from Greece. AM abundance is expressed as: (a) average number of AM per individual among all examined individuals, (b) average number of AM per individual among individuals containing AM, and (c) average number of AM per gram wet weight (w.w.) of gastrointestinal content among individuals containing AM.
SpeciesBoops boopsDicentrarchus labraxSparus aurata
Wild/FarmedWildWildFarmedWildFarmed
TL (cm)23.70 ± 0.8535.30 ± 3.2735.00 ± 2.0427.90 ± 1.5330.50 ± 1.94
RW (kg)0.16 ± 0.020.51 ± 0.130.51 ± 0.070.41 ± 0.070.54 ± 0.05
Number of individuals examined2010101010
Number of individuals containing AM116848
AM frequency of occurrence (%)5560804080
AM number2125361023
AM abundance per individual in all individuals examined a1.10 ± 1.202.50 ± 3.103.60 ± 2.201.00 ± 1.702.30 ± 2.10
AM abundance per individual in individuals containing AM b1.90 ± 1.004.20 ± 2.904.50 ± 1.302.50 ± 1.902.90 ± 1.90
AM abundance per gram weight in individuals containing AM c11.40 ± 6.7021.50 ± 17.6020.00 ± 12.1014.30 ± 13.507.70 ± 5.20
Table 2. Overview of published studies on anthropogenic microparticle (AM) ingestion in Mediterranean fish. Data for species other than those examined in the present study are derived from published sources, as indicated in the references.
Table 2. Overview of published studies on anthropogenic microparticle (AM) ingestion in Mediterranean fish. Data for species other than those examined in the present study are derived from published sources, as indicated in the references.
ReferenceSpeciesNAreaOrigin% With AMAM (Items/Indiv; All)AM
(Items/Indiv; i.c.m.)		AM (Items/g w.w.; i.c.m.)AM Shape (%)AM Size AM Color
This studyB. boops20Cyclades, GreeceWild55%1.10 ± 1.201.90 ± 1.0011.40 ± 6.7086% fibers/14% fragments<0.05 mmBlack
This studyS. aurata10Messolonghi Lagoon, GreeceWild60%1.00 ± 1.702.50 ± 1.9014.30 ± 13.50100% fibers<0.05 mm Black
This studyS. aurata10Rhodes Island, GreeceFarmed80%2.30 ± 2.102.90 ± 1.907.70 ± 5.2087% fibers/13% fragments0.05–1.0 mmBlue
This studyD. labrax10Messolonghi Lagoon, GreeceWild40%2.50 ± 3.104.20 ± 2.9021.50 ± 17.60100% fibers<0.05 mm Black
This studyD. labrax10Rhodes Island, GreeceFarmed80%3.60 ± 2.204.50 ± 1.3020.00 ± 12.1097% fibers/3% fragments0.05–1.0 mmBlack
Anastasopoulou et al. [16]Chelon auratus, S. aurata, Solea solea, Mullus surmuletus, P. erythrinus, S. pilchardus, M. barbatus230 (pooled samples)Slovenian SeaWildNA6.70 ± 3.50NANA75.6% filamentsNANA
Croatian SeaWildNA2.50 ± 0.20NANA97.7% filamentsNANA
NE Ionian SeaWildNA1.70 ± 0.20NANA79% fragmentsNANA
Digka et al. [27]S. pilchardus36Eastern MediterraneanWild47%0.80 ± 0.201.80 ± 0.2034.90 ± 7.9080% fragments/20% fibers0.1–0.5 mmBlue
Digka et al. [27]P. erythrinus19Eastern MediterraneanWild42%0.80 ± 0.201.90 ± 0.2027.80 ± 24.6073% fragments/27% fibers0.1–0.5 mm Blue
Digka et al. [27]M. barbatus25Eastern MediterraneanWild32%0.50 ± 0.201.50 ± 0.3011.20 ± 2.8083% fragments/17% fibers0.1–0.5 mmBlue
Garcia-Garin et al. [17]B. boops34Catalan coast (Barcelona)Wild65%1.68 ± 0.312.59 ± 0.350.83 ± 0.1560% fragments/40% fibers 0.1–0.5 mmBlue
Garcia-Garin et al. [17]B. boops34Catalan coast (Blanes)Wild35%0.50 ± 0.141.42 ± 0.230.20 ± 0.0560% fragments/40% fibers0.1–0.5 mmBlue
Garcia-Garin et al. [17]B. boops34Catalan coast (Cap de Creus MPA)Wild38%0.53 ± 0.141.38 ± 0.180.16 ± 0.0260% fragments/40% fibers0.1–0.5 mmBlack
Tsangaris et al. [19]B. boops884Eastern MediterraneanWild47%NA2.51 ± 0.02NA82% filaments1.0–5.0 mmBlack
Mosconi et al. [28]S. aurata/D. labrax34Eastern MediterraneanFarmed38%0.51 ± 0.781.39 ± 0.65NA68% fibers/32% fragmentsNABlack
Matias et al. [29] D. labrax46Eastern MediterraneanCage aquaculture89%NA3.20 ± 2.300.18 ± 0.1359% fibers/22% fragments 0.1–0.5 mmBlue
50Semi-intensive pond aquaculture94%NA4.20 ± 2.800.53 ± 0.4048% fibers/36% fragments0.1–0.5 mmBlack
55Recirculating aquaculture system (RAS)96%NA4.89 ± 2.500.69 ± 0.42>60% fibers0.1–0.5 mmBlue
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Kostoula, A.; Moschou-Kounopioti, E.; Milatou, N.; Megalofonou, P. Anthropogenic Microparticles in Aquaculture and Wild Fish: A Case Study of Three Commercially Important Species in the Eastern Mediterranean. Fishes 2025, 10, 492. https://doi.org/10.3390/fishes10100492

AMA Style

Kostoula A, Moschou-Kounopioti E, Milatou N, Megalofonou P. Anthropogenic Microparticles in Aquaculture and Wild Fish: A Case Study of Three Commercially Important Species in the Eastern Mediterranean. Fishes. 2025; 10(10):492. https://doi.org/10.3390/fishes10100492

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Kostoula, Aikaterini, Eugenia Moschou-Kounopioti, Niki Milatou, and Persefoni Megalofonou. 2025. "Anthropogenic Microparticles in Aquaculture and Wild Fish: A Case Study of Three Commercially Important Species in the Eastern Mediterranean" Fishes 10, no. 10: 492. https://doi.org/10.3390/fishes10100492

APA Style

Kostoula, A., Moschou-Kounopioti, E., Milatou, N., & Megalofonou, P. (2025). Anthropogenic Microparticles in Aquaculture and Wild Fish: A Case Study of Three Commercially Important Species in the Eastern Mediterranean. Fishes, 10(10), 492. https://doi.org/10.3390/fishes10100492

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