Microplastics in marine mussels, biological effects and human risk of intake: A case study in a multi-stressor environment.

This study documented seasonal levels of microplastics (MPs) and biomarkers (condition index, neurotoxicity, energy, oxidative stress) in mussels (Mytilus galloprovincialis), and water physico-chemical parameters in the Douro estuary (NE Atlantic coast), and estimated the human risk of MP intake (HRI) through mussels. Mussel stress was determined through the Integrated Biomarker Response (IBR). HRI was estimated from mussel MP concentrations and consumer habits. MPs were mainly micro-fibres (72 %) with varied chemical composition. Seasonal MP means (±SEM) in mussels ranged from 0.111 ± 0.044 (spring) to 0.312 ± 0.092 MPs/g (summer). Seasonal variations of mussel stress (IBR: 1.4 spring to 9.7 summer) and MP concentrations were not related. MeO-BDEs, PBDEs, temperature, salinity and other factors likely contributed to mussel stress variation. HRI ranged from 2438 to 2650 MPs/year. Compared to the literature, MP contamination in mussels is low, as well as the human risk of MP intake through their consumption.

Immunotoxicological effects of perfluorooctanesulfonic acid on European seabass are reduced by polyethylene microplastics.

Marine environments receive plastic waste, where it suffers a transformation process into smaller particles. Among them, microplastics (MPs; <5 mm) are ingested by aquatic organisms leading to negative effects on animal welfare. The interactions between MPs, contaminants and organisms are poorly understood. To clarify this issue, European seabass (Dicentrarchus labrax L.) were fed with diets supplemented with 0 (control), polyethylene (PE) MPs (100 mg/kg diet), perfluorooctanesulfonic acid (PFOS, 4.83 µg/kg diet) or PFOS adsorbed to MPs (MPs-PFOS; final concentrations of 4.83 µg and 100 mg of PFOS and MP per kg of feed, respectively). Samples of skin mucus, serum, head-kidney (HK), liver, muscle, brain and intestine were obtained. PFOS levels were high in the liver of fish fed with the PFOS-diet, and markedly reduced when adsorbed to MPs. Compared to the control groups, liver EROD activity did not show any significant changes, whereas brain and muscle cholinesterase activities were decreased in all the groups. The histological and morphometrical study on liver and intestine showed significant alterations in fish fed with the experimental diets. At functional level, all the experimental diets affected the humoral (peroxidase, IgM, protease and bactericidal activities) as well as cellular (phagocytosis, respiratory burst and peroxidase) activities of HK leukocytes, being more marked those effects caused by the PFOS diet. Besides, treatments produced inflammation and oxidative stress as evidenced at gene level. Principal component analysis demonstrated that seabass fed with MPs-PFOS showed more similar effects to MPs alone than to PFOS. Overall, seabass fed with MPs-PFOS diet showed similar or lower toxicological alterations than those fed with MPs or PFOS alone demonstrating the lack of additive effects or even protection against PFOS toxicity.

Are microplastics contributing to pollution-induced neurotoxicity? A pilot study with wild fish in a real scenario.

Pollution-induced neurotoxicity is of high concern. This pilot study investigated the potential relationship between the presence of microplastics (MPs) in the brain of 180 wild fish (Dicentrarchus labrax, Platichthys flesus, Mugil cephalus) from a contaminated estuary and the activity of the acetylcholinesterase (AChE) enzyme. MPs were found in 9 samples (5% of the total), all of them from D. labrax collected in the summer, which represents 45% of the samples of this species collected in that season (20). Seventeen MPs were recovered from brain samples, with sizes ranging from 8 to 96 µm. Polyacrylamide, polyacrylic acid and one biopolymer (zein) were identified by Micro-Raman spectroscopy. Fish with MPs showed lower (p ≤ 0.05) AChE activity than those where MPs were not found. These findings point to the contribution of MPs to the neurotoxicity induced by long-term exposure to pollution, stressing the need of further studies on the topic to increase 'One Health' protection.

Hawksbill Sea Turtle (Eretmochelys imbricata) Blood and Eggs Organochlorine Pesticides Concentrations and Embryonic Development in a Nesting Area (Yucatan Peninsula, Mexico).

Environmental contaminants with chemical origins, such as organochlorine pesticides (OCPs) have major impacts on the health of marine animals, including sea turtles, due to the bioaccumulation of those substances by transference throughout the food chain. The effects of environmental pollution on the health of marine turtles are very important for management strategies and conservation. During recent decades, the south Gulf of Mexico and the Yucatan Peninsula have suffered from increasingly frequent disturbances from continental landmasses, river systems, urban wastewater runoff, port areas, tourism, industrial activities, pesticides from agricultural use, and other pollutants, such as metals, persistent organic pollutants (POPs) and hydrocarbons (from the oil industry activities), which contaminate water and sediments and worsen the environmental quality of the marine ecosystem in this region. In this study, we assessed the concentrations of OCPs in the blood and eggs of 60 hawksbill turtles (Eretmochelys imbricata) nesting at the Punta Xen turtle camp, and their effects on the nesting population's reproductive performance: specifically, maternal transfer and embryonic development were analyzed. Hematologic characteristics, including packed cell volume, white blood cell count, red blood cell count, and haemoglobin levels, and plasma chemistry values, including creatinine, blood urea nitrogen, uric acid, triglyceride, total cholesterol and glucose, were also measured. The general health of the turtles in this study, as well as their levels of urea, serum creatinine, glucose, uric, acid, cholesterol, and triglyceride, fell within normal ranges and was similar to other normal values, which could indicate the turtles' good energy levels and body conditions for nest-building activity, with all of the turtles able to successfully come ashore to nest. All the same, the obtained results also indicate that OCPs affect the nesting and reproductive performance of the hawksbill turtles, as well as their fertility and the development of the population of eggs and reproductive performance, specifically in terms of maternal transference and embryonic development. There were significant differences in the concentrations of OCPs (ΣHCHs and ΣDienes) between maternal blood and eggs, indicating that these chemicals are transferred from nesting females to eggs and, ultimately, to hatchlings. OCPs may, therefore, have an effect on the health and reproductive performance of hawksbill turtles, both in terms of their fertility and egg development. Conservation strategies need to be species-specific, due to differences in feeding, and address the reasons for any decline, focusing on regional assessments. Thus, accurate and comparable monitoring data are necessary, which requires the standardization of monitoring protocols.

Occurrence and risk assessment of endocrine-disrupting compounds in fish muscle: The case study of the Douro River estuary (North East Atlantic Ocean).

This work was focused on evaluating the occurrence of endocrine-disrupting compounds (EDCs) in fish muscles, such as bisphenol analogues, personal care products, including some UV-filters and musk fragrances, and selected pesticides. A total of 238 muscle samples of three fish species (Platichthys flesus, Mugil cephalus, and Dicentrarchus labrax) of an important estuary ending in the North East Atlantic Ocean (Douro River estuary, Portugal), in different seasons of the year, was analysed. M. cephalus was the species with a greater occurrence of contaminants, mainly in fishes collected during the spring. Bisphenol A was the main bisphenol analogue detected with an average content of 23.1 µg/kg wet weight (w.w.). Among chemicals used in personal care products, galaxolide, tonalide, and 2-ethylhexyl salicylate, were the most often found, being present in 15.9% (2.2-538.4 µg/kg w. w.), 4.6% (1.1-57.8 µg/kg w. w.) and 3.4% (3.9-56.1 µg/kg w. w.) of the samples, respectively. About 14% of the samples contained residues of at least one pesticide, being alachlor, aldrin, p,p'-DDT, permethrin, and prochloraz the main chemicals observed, varying from 0.1 µg/kg w. w. (p,p'-DDT) to 37.8 µg/kg w. w. (prochloraz). The daily intake estimates of the most frequent EDCs found in the fish muscles suggested that there are no health concerns, based on the recommended weekly consumption of fish for adults.

Occurrence and seasonal variation of several endocrine disruptor compounds (pesticides, bisphenols, musks and UV-filters) in water and sediments from the estuaries of Tagus and Douro Rivers (NE Atlantic Ocean coast).

Exposure of aquatic environments to emerging contaminants is a global issue, special relevant in many estuaries due to impacts from anthropogenic activity. The aim of this work was to evaluate thirty-seven endocrine disruptor chemicals (EDCs) from four different classes (pesticides, bisphenols, polycyclic musks and UV-filters) in water and sediment samples collected during one-year in the estuaries of Tagus and Douro Rivers located into the NE Atlantic Ocean coast. EDCs analysis was achieved afterward validation of a gas-chromatography mass spectrometry (GC-MS) method using Dispersive Liquid-Liquid Microextraction (DLLME) as extraction procedure for water samples, and Quick, Easy, Cheap, Efficient, Rugged and Safe (QuEChERS) combined with DLLME for sediments. Tagus estuary presented higher levels of contamination with pesticide residues and bisphenols (BPs) than the Douro estuary in both water and sediment samples. Contrariwise, levels and frequency of polycyclic musks (PCMs) and UV-filters (UVF) were slightly higher in Douro estuary. Levels of pesticide residues in both sediment and water samples, and levels of PCMs and UVF in water samples were higher in warmer seasons (summer and spring) than in colder ones (winter and autumn). The opposite was found in what respect levels of BPs in water and sediment samples, and PCMs and UVF levels in sediment samples. Although the levels found for each contaminant are low, usually in the order of a few ng/mL(g), the presence of a high number of toxic compounds is a source of concern and requires constant monitoring.

Microplastics in fishes from an estuary (Minho River) ending into the NE Atlantic Ocean.

Wild fish (Cyprinus carpio, Mugil cephalus, Platichthys flesus) from an estuary of the NE Atlantic coast were investigated for plastic contamination (N = 128). From the 1289 particles recovered from fish samples, 883 were plastics. Among these, 84% were fibres and 97% were microplastics. Thirty-six polymers were identified. The number of microplastics (mean ± SD) per individual fish (MP/fish) was 8 ± 6 in C. carpio, 10 ± 9 in M. cephalus and 2 ± 2 in P. flesus. The means of MP/fish per body site were 6 ± 7 in gastrointestinal tract, 0.5 ± 1.1 in gills, 0.3 ± 0.7 in liver and 0.6 ± 1.2 in muscle samples. A few large fibres in liver (≤ 4841 µm) and muscle (≤ 5810 µm) samples were found. The results evidence the existence of high fish contamination by microplastics and reinforce the need of further research on plastic pollution in estuaries.

Polybrominated diphenyl ethers and their methoxylated congeners in Douro river estuary biota: Seasonal occurrence and risk assessment.

Especially added on many industrial and domestic products as flame retardants (FRs), polybrominated diphenyl ethers (PBDEs) are among the chemicals of high environmental concern because of their potential harmfulness for environmental and human health. Seafood consumption is considered the main source of PBDEs and their methoxylated congeners (MeO-BDEs) for humans. The present study aims to investigate the seasonal occurrence of six PBDEs and eight MeO-BDEs congeners using Douro river biota (different trophic levels) as sentinels, as well as to evaluate the human exposure risk to PBDEs through seafood consumption. Biota samples (n = 273) were collected from one of the most important Portuguese estuaries in the north-western coast of Portugal at four different seasons (2019-2020). The analyses were performed by an environmental-friendly extraction procedure followed by Gas Chromatography coupled to a triple quadrupole detector (GC-MS/MS). PBDEs were detected in all seafood samples analysed, with means ranging from 0.02 ng g-1 ww (flounder in autumn) to 3.75 ng g-1 ww (mussel in winter). Levels of lower-brominated PBDE congeners were significantly higher than higher-brominated ones in all seasons (p < 0.01). MeO-BDEs ranged from 0.001 ng g-1 ww (grey mullet in summer) to 5.66 ng g-1 ww (green crab in spring). Crabs and mussels presented the highest means of PBDEs and MeO-BDEs. Regarding the health risk assessment of the studied PBDE congeners (47, 99, and 153), consumption of Douro river fish is not a case of concern for consumers.

Neurotoxicity, Behavior, and Lethal Effects of Cadmium, Microplastics, and Their Mixtures on Pomatoschistus microps Juveniles from Two Wild Populations Exposed under Laboratory Conditions-Implications to Environmental and Human Risk Assessment.

Microplastics (MPs) were found to modulate the toxicity of other pollutants but the knowledge on the topic is still limited. The goals of this study were to investigate the short-term toxicity of cadmium (Cd) to wild Pomatochistus microps juveniles, the potential modulation of acute Cd toxicity by 1-5 µm polyethylene MPs in this species, and possible differences of sensitivity to Cd and MPs-Cd mixtures between juveniles from two distinct wild populations. Juveniles were collected in the estuaries of Minho (M-est) and Lima (L-est) Rivers (NW Portugal). One 96 h bioassay with M-est juveniles and another one with L-est juveniles were carried out in laboratory conditions. Each bioassay had 12 treatments: control, 5 Cd concentrations, 1 MPs concentration, and 5 MPs-Cd mixtures. No significant differences in Cd-induced mortality between juveniles from distinct estuaries or between juveniles exposed to Cd alone and those exposed to MPs-Cd mixtures were found. The total 96h LC10 and LC50 of Cd alone were 2 mg/L (95% CI: 0-4 mg/L) and 8 mg/L (95% CI: 2-17 mg/L), respectively. Cd alone significantly decreased the post-exposure predatory performance (PEPP) of M-est (≥6 mg/L) and L-est juveniles (≥3 mg/L), and acetylcholinesterase (AChE) activity of M-est juveniles (13 mg/L). MPs alone (0.14 mg/L) significantly reduced the PEPP and AChE activity of L-est juveniles but not of M-est juveniles. MPs-Cd mixtures (3-13 mg/L of Cd + 0.14 mg/L of MPs) significantly inhibited the PEPP of juveniles from both estuaries and AChE of L-est estuary juveniles but not of M-est juveniles. Evidences of toxicological interactions, namely antagonism, between MPs and Cd were found. Overall, the results indicate that MPs modulated the sub-lethal toxic effects of Cd in wild P. microps juveniles, especially neurotoxicity. Moreover, the environmental conditions of the natural habitats to which juveniles were exposed during pre-developmental phases influence the sub-lethal toxicity of Cd, MPs, and their mixtures. The implications to environmental and human risk assessment are discussed and further research is needed.

Organochlorines Contaminants in Eggs of Hawksbill (Eretmochelys imbricata) and Green Sea Turtles (Chelonia mydas) from Mexico coast.

The investigation of organochlorine pesticides (OCPs) levels in sea turtles is an important issue in conservation research, due to the harmful effects of these chemicals. In the present study, OCPs concentrations were determined in the eggs of two sea turtle species (Eretmochelys imbricata and Chelonia mydas) collected from the Punta Xen and Isla Aguada (Mexican coast) in 2014 and 2015. Concentrations of 20 OCPs were analysed, including isomers of hexachlorocyclohexane, aldrin, chlordanes, endosulfans, methoxychlor, DDTs, and heptachlor. From the group of contaminants considered (analysed as families), the results revealed higher concentrations of ΣHCH and ΣDienes on both selected species. We analysed the relationship between turtle size and the OCPs concentrations; no correlation was found between the size of the female and concentrations in the eggs. In addition, principal component analysis indicated pattern differences between species and years, in good agreement with concentrations differences.

Oxidative stress biomarkers and organochlorine pesticides in nesting female hawksbill turtles Eretmochelys imbricata from Mexican coast (Punta Xen, Mexico).

Because of their vulnerable population status, assessing exposure levels and impacts of toxicants on the health status of Gulf of Mexico marine turtle populations is essential, and this study was aimed to obtain baseline information on oxidative stress indicators in hawksbill sea turtle (Eretmochelys imbricata). In order to evaluate the health status of sea turtles and the effect of organochlorine compounds (OC) in the southern part of the Gulf of Mexico, we searched for relationships between carapace size and the activity of antioxidant enzymes in the blood of the hawksbill sea turtle. The level of oxidative stress biomarkers such as the enzymes catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), glutathione S-transferase (GST), and acetylcholinesterase (Ache) in the hawksbill sea turtle was analysed during nesting season in the years 2014-2015 at Punta Xen (Campeche, Mexico). The results of this study provide insight into data of antioxidant enzyme activities in relation to contaminant OCPs in hawksbill sea turtles and the possible health impacts of contaminant in sea turtles.

Uptake and effects of the antimicrobial florfenicol, microplastics and their mixtures on freshwater exotic invasive bivalve Corbicula fluminea.

Microplastics and antimicrobials are widely spread environmental contaminants and more research on their toxicity is needed. The uptake and effects of the antimicrobial florfenicol, microplastics, and their mixtures on Corbicula fluminea were investigated. Bivalves were exposed for 96h to florfenicol (1.8 and 7.1mg/l), microplastics (0.2 and 0.7mg/l), or mixtures of the two substances. After 96h, all bivalves exposed to antimicrobial treatments had florfenicol in their body (e.g. 2±1µg/g). Microplastics were found in the gut, lumen of the digestive gland, connective tissue, hemolymphatic sinuses, and gills surface of animals. Florfenicol caused a significant inhibition of cholinesterase (ChE) activity (~32%). Animals exposed to 0.2mg/l of microplastics showed ChE activity inhibition (31%), and no other significant alterations. Mixtures caused feeding inhibition (57-83%), significant ChE inhibition (44-57%) and of isocitrate dehydrogenase activity, and increased anti-oxidant enzymes activity and lipid peroxidation levels. Overall, the results indicate that C. fluminea take up florfenicol and microplastics from the water and accumulated or at least retained it in their body for some time; both florfenicol (low ppm range) and microplastics (ppb range) were toxic to C. fluminea, with mixtures containing florfenicol and microplastics being more toxic. Thus, the risk of exposure and toxic effects of florfenicol to C. fluminea and other bivalves, and its predators increase in ecosystems contaminated with the antimicrobial and microplastics, as well as to humans consuming contaminated species from these ecosystems.