Microplastic Pollution in the Gastrointestinal Tract and Gills of Some Teleost and Sturgeon Fish from the Caspian Sea, Northern Iran.

The increasing microplastic pollution in the marine environment has raised global concern. The main risk of microplastics in aquatic ecosystem is their bioaccumulation in aquatic organisms. A few studies have reported microplastic pollution in the digestive system of Caspian Sea fish species, but there is no research on sturgeon species, nor on fish gills. We investigated the occurrence of microplastics in the gastrointestinal tract (GIT) and gills of 62 specimens belonging to four species including three teleosts (Cyprinus carpio, Rutilus kutum, and Chelon aurata) and one sturgeon (Acipenser persicus, a valuable endangered species) from the Caspian Sea between January and March 2022. Fish tissues were removed, exposed for 24 h to 10% KOH, and then dried on filter paper. Particles were observed under a stereomicroscope and analyzed by Raman microspectrometry, scanning electron microscopy, and energy-dispersive spectroscopy. A total of 91 microplastics were detected in the GIT (average of 1.46 ± 1.17 items/individual) and 63 microplastics in the gills (average of 1.01 ± 0.62 items/individual). A significant correlation was not found between the number of microplastics found in both tissues and fish body length, body weight, GIT weight, and gill weight (p > 0.05), except between microplastics isolated from gills and gill weight in C. carpio (rs = 0.707, p = 0.022). The abundance of microplastics in fish followed the order of A. persicus > C. aurata > R. kutum > C. carpio. The microplastics were in the size range of 45 to 5000 µm, with particles of 300 to 1000 µm being the most prevalent; 74.68% of the particles were shaped like fibers, 30.53% were red, and 70.6% were composed of nylon polymer. Environ Toxicol Chem 2023;42:2453-2465. © 2023 SETAC.

Microplastics in different tissues of some commercially important fish species from Anzali Wetland in the Southwest Caspian Sea, Northern Iran.

The aim of this study was to investigate the occurrence of microplastics (MPs) in the gastrointestinal (GI) tract, muscle and gonads of 193 individuals of nine commercially-important fish species from Anzali Wetland between May and July 2018. Tissues were removed, digested by 10% KOH for 24 h at 60 °C, filtered on cellulose nitrate filter and then dried. MPs were found in all tissues. There were significant differences between the numbers of isolated particles from GI tract and control groups (procedural blank containing Milli Q water without tissue), while their abundance in the muscle and gonads did not display significant differences with control groups. Higher abundances of MPs were recorded in omnivorous fish (2.26 ± 2.93 items/individual) than carnivorous species (1.10 ± 1.10 items/individual) (P < 0.05). MPs were ranged from 20 to 4800 µm in size and were dominated by fiber-like in shape, polyethylene in polymer and black in color.

Identification and distribution of microplastics in the sediments and surface waters of Anzali Wetland in the Southwest Caspian Sea, Northern Iran.

The occurrence, abundance and distribution of microplastics (MPs) were studied in sediments and surface waters of the Anzali coastal wetland, southwest of the Caspian Sea. Sediment and surface water samples were taken at 11 and 6 stations in June 2018 and January 2019 respectively. The abundances of MPs in sediment samples were 140-2820 and 113-3690 items/kg dry weight while in surface waters were 0.40-4.41 and 0.19-2.85 items/m3 in June and January respectively. Fiber was the most common shape, followed by fragment and film. Red, black and blue were the most frequent colors. Particles 1-2 mm were the dominant size. SEM/EDS and ATR-FTIR analyses were used to identify MPs. Polypropylene, polyethylene and polyester were the most frequently found polymers. Our results highlighted the widespread dispersion of MPs in Anzali Wetland which in turn might be a pathway of MPs pollution transporter to the adjacent and connected Caspian Sea.

Distribution and Characterization of Microplastics in Surface Waters and the Southern Caspian Sea Coasts Sediments.

The Caspian Sea is the largest enclosed aquatic ecosystem in the world. The combinations of the toxic pollutants with microplastics endanger the Caspian Sea ecosystem. In this work, the distribution of microplastics was studied in surface waters and southern Caspian Sea coasts sediments. The samples were collected from eight stations, including the Tonakabon, Chalos, Nowshahr, Noor, Mahmood Abad, Babolsar, Sari, and Neka coasts. The average concentrations of microplastics in the coastal waters and sediments were 34,490 particles per km2 and 210 particles per kg, respectively. Isolated microplastics were characterized using ATR-FTIR and energy dispersive X-ray (EDS) techniques. The samples exhibited a strong carbon peak in the EDS spectra, which was screened as microplastic particles. The microplastics were mainly fragments and foams and identified as polyethylene, polypropylene, and polystyrene by means of ATR-FTIR spectra. This is the first study to determine the distribution of microplastics in southern Caspian coastal regions.

Arsenic-containing hydrocarbons and arsenic-containing fatty acids: Transfer across and presystemic metabolism in the Caco-2 intestinal barrier model.

SCOPE: Arsenic-containing hydrocarbons (AsHCs) and arsenic-containing fatty acids (AsFAs) represent two classes of arsenolipids occurring naturally in marine food. Toxicological data are yet scarce and an assessment regarding the risk to human health has not been possible. Here, we investigated the transfer and presystemic metabolism of five arsenolipids in an intestinal barrier model. METHODS AND RESULTS: Three AsHCs and two AsFAs were applied to the Caco-2 intestinal barrier model. Thereby, the short-chain AsHCs reached up to 50% permeability. Transport is likely to occur via passive diffusion. The AsFAs showed lower intestinal bioavailability, but respective permeabilities were still two to five times higher as compared to arsenobetaine or arsenosugars. Interestingly, AsFAs were effectively biotransformed while passing the in vitro intestinal barrier, whereas AsHCs were transported to the blood-facing compartment essentially unchanged. CONCLUSION: AsFAs can be presystemically metabolised and the amount of transferred arsenic is lower than that for AsHCs. In contrast, AsHCs are likely to be highly intestinally bioavailable to humans. Since AsHCs exert strong toxicity in vitro and in vivo, toxicity studies with experimental animals as well as a human exposure assessment are needed to assess the risk to human health related to the presence of AsHCs in seafood.

Arsenolipids in oil from blue whiting Micromesistius poutassou--evidence for arsenic-containing esters.

Arsenic-containing lipids in the oil from the blue whiting fish (Micromesistius poutassou) were separated into three broad polarity groups and investigated by HPLC and mass spectrometry. A total of 11 arsenolipids including 4 new compounds were identified. The polar lipid fraction constituting 24% of the total arsenolipid content (which totalled 2.16 µg As/g) contained four known dimethylarsinoyl fatty acids and three known dimethylarsinoyl hydrocarbons. The less polar fraction (ca 30% of the total arsenolipids) contained four new dimethylarsinoyl hydrocarbons with chain lengths 22-30 carbons, in addition to more complex arsenicals that hydrolysed to known dimethylarsinoyl fatty acids suggesting they were conjugated carboxylic acids, presumably esters. The rest of the lipid-soluble arsenic (ca 45% of the total) remained in the non-polar fraction together with the bulk of the fish oil lipids, a complex mixture of compounds that precluded identification of the small amounts of arsenolipids.

Synthesis and Characterization of Arsenolipids: Naturally Occurring Arsenic Compounds in Fish and Algae.

Arsenic-containing lipids (arsenolipids) are natural products present in fish and algae. Because these compounds occur in foods, there is considerable interest in their human toxicology. We report the synthesis and characterization of seven arsenic-containing lipids, including six natural products. The compounds comprise dimethylarsinyl groups attached to saturated long-chain hydrocarbons (three compounds), saturated long-chain fatty acids (two compounds), and monounsaturated long chain fatty acids (two compounds). The arsenic group was introduced through sodium dimethylarsenide or bis(dimethylarsenic) oxide. The latter route provided higher and more reproducible yields, and consequently, this pathway was followed to synthesize six of the seven compounds. Mass spectral properties are described to assist in the identification of these compounds in natural samples. The pure synthesized arsenolipids will be used for in vitro experiments with human cells to test their uptake, biotransformation, and possible toxic effects.

In vitro intestinal bioavailability of arsenosugar metabolites and presystemic metabolism of thio-dimethylarsinic acid in Caco-2 cells.

Whereas inorganic arsenic is classified as a human carcinogen, risks to human health related to the presence of arsenosugars in marine food are still unclear. Since studies indicate that human inorganic arsenic metabolites contribute to inorganic arsenic induced carcinogenicity, a risk assessment for arsenosugars should also include a toxicological characterization of their respective metabolites. Here we assessed intestinal bioavailability of the human arsenosugar metabolites oxo-DMAA(V), thio-DMAA(V), oxo-DMAE(V), thio-DMAE(V) and thio-DMA(V) in relation to arsenite in the Caco-2 intestinal barrier model. Whereas arsenite and thio-DMA(V) caused barrier disruption at concentrations ≥10 µM, all other metabolites did not cause a barrier leakage, even when applied at 50 times higher concentrations than arsenite and thio-DMA(V). The transfer studies point to a strong intestinal bioavailability of thio-DMA(V) and thio-DMAE(V), whereas oxo-DMAA(V), thio-DMAA(V) and oxo-DMAE(V) passed the in vitro intestinal barrier only to a very small extent. Detailed influx and efflux studies indicate that arsenite and thio-DMA(V) cross the intestinal barrier most likely by passive diffusion (paracellular) and facilitated (transcellular) transport. LC-ICP-QMS based arsenic speciation studies during the transfer experiments demonstrate transfer of thio-DMA(V) itself across the intestinal barrier and suggest metabolism of thio-DMA(V) using the in vitro intestinal barrier model to its oxygen-analogue DMA(V). In the case of arsenite no metabolism was observed. In summary the two arsenosugar metabolites thio-DMA(V) and thio-DMAE(V) showed intestinal bioavailability similar to that of arsenite, and about 10-fold higher than that reported for arsenosugars (Leffers et al., Mol. Nutr. Food Res., 2013, DOI: 10.1002/mnfr.201200821) in the same in vitro model. Thus, a presystemic metabolism of arsenosugars might strongly impact arsenic intestinal bioavailability after arsenosugar intake and should therefore be considered when assessing the risks to human health related to the consumption of arsenosugar-containing food.

In vitro toxicological characterization of two arsenosugars and their metabolites.

SCOPE: In their recently published Scientific Opinion on Arsenic in Food, the European Food Safety Authority concluded that a risk assessment for arsenosugars is currently not possible, largely because of the lack of relevant toxicological data. To address this issue, we carried out a toxicological in vitro characterization of two arsenosugars and six arsenosugar metabolites. METHODS AND RESULTS: The highly pure synthesized arsenosugars, DMA(V) -sugar-glycerol and DMA(V) -sugar-sulfate, investigated in this study, as well as four metabolites, oxo-dimethylarsenoacetic acid (oxo-DMAA(V) ), oxo-dimethylarsenoethanol (oxo-DMAE(V) ), thio-DMAA(V) and thio-DMAE(V) , exerted neither cytotoxicity nor genotoxicity up to 500 µM exposure in cultured human bladder cells. However, two arsenosugar metabolites, namely dimethyl-arsinic acid (DMA(V) ) and thio-dimethylarsinic acid (thio-DMA(V) ), were toxic to the cells; thio-DMA(V) was even slightly more cytotoxic than arsenite. Additionally, intestinal bioavailability of the arsenosugars was assessed applying the Caco-2 intestinal barrier model. The observed low, but significant transfer rates of the arsenosugars across the barrier model provide further evidence that arsenosugars are intestinally bioavailable. CONCLUSION: In a cellular system that metabolizes arsenosugars, cellular toxicity likely arises. Thus, in strong contrast to arsenobetaine, arsenosugars cannot be categorized as nontoxic for humans and a risk to human health cannot be excluded.

Arsenic-containing lipids are natural constituents of sashimi tuna.

Arsenic occurs naturally in many types of seafood as water- and fat-soluble organoarsenic compounds. Although water-soluble compounds have been well characterized, the fat-soluble compounds, so-called arsenolipids, have until recently remained unknown. We report that sashimi-grade tuna fish, with a total arsenic content of 5.9 microg of As/g dry mass, contains approximately equal quantities of water- and fat-soluble arsenic. The water-soluble arsenic comprised predominantly arsenobetaine (>95%) with a trace of dimethylarsinate. Two fat-soluble compounds, which together accounted for about 40% of the lipid-arsenic, were isolated and characterized. The first was identified as 1-dimethylarsinoylpentadecane [(CH(3))(2)As(O)(CH(2))(14)CH(3)] by comparison of HPLC/mass spectrometric data and accurate mass data with those of an authenticated synthesized standard. The second arsenolipid was postulated as 1-dimethylarsinoyl all-cis-4,7,10,13,16,19-docosahexane from mass spectrometric data and analogy with non-arsenic-containing lipids found in fish. The remaining fat-soluble arsenic consisted of less polar arsenolipids of currently unknown structure. This is the first identification of arsenolipids in commonly consumed seafood.

Identification of arsenolipids with GC/MS.

Arsenic-containing hydrocarbons have recently been reported as natural constituents of fish oil. We report a simple method for determining these compounds by GC/MS. Application of the methodology will delineate the distribution of these novel arsenic compounds in foods, and facilitate an assessment of the toxicological implications.

Arsenic-containing hydrocarbons: natural compounds in oil from the fish capelin, Mallotus villosus.

Arsenic-containing hydrocarbons have been identified for the first time as natural components of fish oil.