Speciation analysis and toxicity evaluation of arsenolipids-an overview focusing on sea food.

Arsenic, which can be divided into inorganic and organic arsenic, is a toxic metalloid that has been identified as a human carcinogen. A common source of arsenic exposure in seafood is arsenolipid, which is a complex structure of lipid-soluble organic arsenic compounds. At present, the known arsenolipid species mainly include arsenic-containing fatty acids (AsFAs), arsenic-containing hydrocarbons (AsHCs), arsenic glycophospholipids (AsPLs), and cationic trimethyl fatty alcohols (TMAsFOHs). Furthermore, the toxicity between different species is unique. However, the mechanism underlying arsenolipid toxicity and anabolism remain unclear, as arsenolipids exhibit a complex structure, are present at low quantities, and are difficult to extract and detect. Therefore, the objective of this overview is to summarize the latest research progress on methods to evaluate the toxicity and analyze the main speciation of arsenolipids in seafood. In addition, novel insights are provided to further elucidate the speciation, toxicity, and anabolism of arsenolipids and assess the risks on human health.

The impact of nano/micro-plastics toxicity on seafood quality and human health: facts and gaps.

Contamination of the food and especially marine environment with nano/micro-plastic particles has raised serious concern in recent years. Environmental pollution and the resulting seafood contamination with microplastic (MP) pose a potential threat to consumers. The absorption rate of the MP by fish is generally considered low, although the bioavailability depends on the physical and chemical properties of the consumed MP. The available safety studies are inconclusive, although there is an indication that prolonged exposure to high levels of orally administered MP can be hazardous for consumers. This review details novel findings about the occurrence of MP, along with its physical and chemical properties, in the marine environment and seafood. The effect of processing on the content of MP in the final product is also reviewed. Additionally, recent findings regarding the impact of exposure of MP on human health are discussed. Finally, gaps in current knowledge are underlined, and the possibilities for future research are indicated in the review. There is an urgent need for further research on the absorption and bioavailability of consumed MP and in vivo studies on chronic exposure. Policymakers should also consider the implementation of novel legislation related to MP presence in food.

Contribution of child ABC-transporter genetics to prenatal MeHg exposure and neurodevelopment.

BACKGROUND: There is emerging evidence that exposure to prenatal methylmercury (MeHg) from maternal fish consumption during pregnancy can differ between individuals due to genetic variation. In previous studies, we have reported that maternal polymorphisms in ABC-transporter genes were associated with maternal hair MeHg concentrations, and with children's early neurodevelopmental tests. In this study, we add to these findings by evaluating the contribution of genetic variation in children's ABC-transporter genes to prenatal MeHg exposure and early child neurodevelopmental tests. METHODS: We genotyped six polymorphisms (rs2032582, rs10276499 and rs1202169 in ABCB1; rs11075290 and rs215088 in ABCC1; rs717620 in ABCC2) in DNA from cord blood and maternal blood of the Seychelles Child Development Study Nutrition Cohort 2. We determined prenatal MeHg exposure by measuring total mercury (Hg) in cord blood by atomic fluorescence spectrometry. We assessed neurodevelopment in children at approximately 20 months using the Bayley Scales of Infant Development (BSID-II). We used linear regression models to analyze covariate-adjusted associations of child genotype with cord MeHg and BSID-II outcomes (Mental Developmental and Psychomotor Developmental Indexes). We also evaluated interactions between genotypes, cord MeHg, and neurodevelopmental outcomes. All models were run with and without adjustment for maternal genotype. RESULTS: Of the six evaluated polymorphisms, only ABCC1 rs11075290 was associated with cord blood MeHg; children homozygous for the T-allele had on average 29.99 µg/L MeHg in cord blood while those homozygous for the C-allele had on average 38.06 µg/L MeHg in cord blood (p < 0.001). No polymorphisms in the children were associated with either subscale of the BSID. However, the association between cord MeHg and the Mental Developmental Index (MDI) of the BSID differed significantly across the three genotypes of ABCB1 rs10276499 (2df F-test, p = 0.045). With increasing cord MeHg, the MDI decreased (slope=-0.091, p = 0.014) among children homozygous for the rare C-allele. CONCLUSIONS: These findings support the possibility that child ABC genetics might influence prenatal MeHg exposure.

Fish Cytolysins in All Their Complexity.

The majority of the effects observed upon envenomation by scorpaenoid fish species can be reproduced by the cytolysins present in their venoms. Fish cytolysins are multifunctional proteins that elicit lethal, cytolytic, cardiovascular, inflammatory, nociceptive, and neuromuscular activities, representing a novel class of protein toxins. These large proteins (MW 150-320 kDa) are composed by two different subunits, termed α and ß, with about 700 amino acid residues each, being usually active in oligomeric form. There is a high degree of similarity between the primary sequences of cytolysins from different fish species. This suggests these molecules share similar mechanisms of action, which, at least regarding the cytolytic activity, has been proved to involve pore formation. Although the remaining components of fish venoms have interesting biological activities, fish cytolysins stand out because of their multifunctional nature and their ability to reproduce the main events of envenomation on their own. Considerable knowledge about fish cytolysins has been accumulated over the years, although there remains much to be unveiled. In this review, we compiled and compared the current information on the biochemical aspects and pharmacological activities of fish cytolysins, going over their structures, activities, mechanisms of action, and perspectives for the future.

New Method for Simultaneous Arsenic and Selenium Speciation Analysis in Seafood and Onion Samples.

This paper presents a new method for the simultaneous speciation analysis of arsenic (As(III)-arsenite, As(V)-arsenate, DMA-dimethylarsinic acid, MMA-methylarsonic acid, and AsB-arsenobetaine) and selenium (Se(IV)-selenite, Se(VI)-selenate, Se-Methionine, and Se-Cystine), which was applied to a variety of seafood and onion samples. The determination of the forms of arsenic and selenium was undertaken using the High-Performance Liquid Chromatography Inductively Coupled Plasma Mass Spectrometry (HPLC-ICP-MS) analytical technique. The separation of both organic and inorganic forms of arsenic and selenium was performed using two analytical columns: an anion exchange column, Dionex IonPac AS22, containing an alkanol quaternary ammonium ion, and a double bed cation-anion exchange guard column, Dionex Ion Pac CG5A, containing, as a first layer, fully sulfonated latex for cation exchange and a fully aminated layer for anion exchange as the second layer. The ammonium nitrate, at pH = 9.0, was used as a mobile phase. The method presented here allowed us to separate the As and Se species within 10 min with a suitable resolution. The applicability was presented with different sample matrix types: seafood and onion.

Identification of Fish Species and Toxins Implicated in a Snapper Food Poisoning Event in Sabah, Malaysia, 2017.

In the coastal countries of Southeast Asia, fish is a staple diet and certain fish species are food delicacies to local populations or commercially important to individual communities. Although there have been several suspected cases of ciguatera fish poisoning (CFP) in Southeast Asian countries, few have been confirmed by ciguatoxins identification, resulting in limited information for the correct diagnosis of this food-borne disease. In the present study, ciguatoxin-1B (CTX-1B) in red snapper (Lutjanus bohar) implicated in a CFP case in Sabah, Malaysia, in December 2017 was determined by single-quadrupole selected ion monitoring (SIM) liquid chromatography/mass spectrometry (LC/MS). Continuous consumption of the toxic fish likely resulted in CFP, even when the toxin concentration in the fish consumed was low. The identification of the fish species was performed using the molecular characterization of the mitochondrial cytochrome c oxidase subunit I gene marker, with a phylogenetic analysis of the genus Lutjanus. This is the first report identifying the causative toxin in fish-implicated CFP in Malaysia.

Mycotoxin Identification and In Silico Toxicity Assessment Prediction in Atlantic Salmon.

The present study aimed to identify mycotoxins in edible tissues of Atlantic salmon (Salmo salar) using liquid chromatography coupled to hybrid quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). After using a non-targeted screening approach and a home-made spectral library, 233 mycotoxins were analyzed. Moreover, the occurrence of mycotoxins in fish filets was evaluated, and their potential toxicity was predicted by in silico methods. According to the obtained results, forty mycotoxins were identified in analyzed salmon samples, the predominant mycotoxins being enniatins (also rugulosin and 17 ophiobolins), commonly found in cereals and their by-products. Thus, mycotoxin carry-over can occur from feed to organs and edible tissues of cultivated fish. Moreover, the toxicity of detected mycotoxins was predicted by the in silico webserver ProTox-II, highlighting that special attention must be paid to some less reported mycotoxins due to their toxic predicted properties.

Neuro-2a cell-based assay for toxicity equivalency factor - proposal and evaluation in Chilean contaminated shellfish samples.

There are two official PSP detection methods (mouse bioassay and HLPC-FLD) and a number of alternative methods. Ethical considerations have led to regulations being adopted in some countries that limit or prohibit the application of mouse bioassay. Analytical methodologies (e.g. HPLC-FLD or LC-MSMS) have the disadvantages of not being able to detect new toxins or analogues or reflecting the overall toxicity of the sample. In addition, they require highly trained personnel and expensive equipment, which are not always available. In this work, we have evaluated a method based on the Neuro-2a cell-based assay to detect substances that inhibit voltage-dependent sodium channels (Manger's method). We tested PSP standards and natural samples contaminated with PSP. Here we demonstrate that the adapted Manger's method is suitable for calculating Toxicity Equivalency Factors (TEF) for STX-analogues. The method was shown to be useful for screening contaminated natural samples in concentrations above the regulatory limit for these toxins (80 µg STX equivalents/100 g shellfish). We were able to detect PSP in 19 natural mollusc samples from South Chile despite the presence of other marine toxins. These preliminary results suggest that the method could be used as a first step in screening programmes.

Associations of Exposure to Perfluoroalkyl Substances With Thyroid Hormone Concentrations and Birth Size.

BACKGROUND: Adequate thyroid function during pregnancy is essential for optimal fetal growth. Gestational exposure to perfluoroalkyl substances (PFAS) can negatively affect birth size and disrupt maternal and neonatal thyroid function, although the interrelationship is unclear. OBJECTIVE: We aimed to quantify the associations between maternal serum-PFAS concentrations and birth weight, birth length, and cranial circumference. We also aimed to estimate associations between PFAS and thyroid hormone (TH) concentrations, thereby elucidating whether THs potentially mediate the associations between PFAS concentrations and birth size. METHODS: We studied a population-based prospective cohort of 172 mother-singleton pairs from the Faroe Islands. Twelve PFAS were measured in maternal serum obtained at 34 weeks of gestation. THs were measured in maternal and cord serum. Associations between PFAS concentrations and birth size and TH concentrations were estimated using multivariable linear regressions. Sex-stratified analyses along with a mediation analysis were performed to estimate potential mediating effects of THs in the association between PFAS and birth outcomes. RESULTS: Several PFASs were negatively associated with birth weight, length, and head circumference, and a general positive association between maternal serum-PFASs and cord serum-thyroid-stimulating hormone (TSH; also known as thyrotropin) was found. For instance, a doubling in perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) was associated with a 53% (95% CI, 18%-99%) and 40% (95% CI, 8%-81%) increases in TSH concentrations, respectively. There was little evidence of sexually dimorphic associations. Overall, THs were not found to mediate associations between PFASs and birth size. CONCLUSION: In this study, several PFASs were negatively associated with birth size and increased THs; however, this did not explain lower birth weight among children exposed to PFAS.

Biological Effects of the Azaspiracid-Producing Dinoflagellate Azadinium dexteroporum in Mytilus galloprovincialis from the Mediterranean Sea.

Azaspiracids (AZAs) are marine biotoxins including a variety of analogues. Recently, novel AZAs produced by the Mediterranean dinoflagellate Azadinium dexteroporum were discovered (AZA-54, AZA-55, 3-epi-AZA-7, AZA-56, AZA-57 and AZA-58) and their biological effects have not been investigated yet. This study aimed to identify the biological responses (biomarkers) induced in mussels Mytilus galloprovincialis after the bioaccumulation of AZAs from A. dexteroporum. Organisms were fed with A. dexteroporum for 21 days and subsequently subjected to a recovery period (normal diet) of 21 days. Exposed organisms accumulated AZA-54, 3-epi-AZA-7 and AZA-55, predominantly in the digestive gland. Mussels' haemocytes showed inhibition of phagocytosis activity, modulation of the composition of haemocytic subpopulation and damage to lysosomal membranes; the digestive tissue displayed thinned tubule walls, consumption of storage lipids and accumulation of lipofuscin. Slight genotoxic damage was also observed. No clear occurrence of oxidative stress and alteration of nervous activity was detected in AZA-accumulating mussels. Most of the altered parameters returned to control levels after the recovery phase. The toxic effects detected in M. galloprovincialis demonstrate a clear biological impact of the AZAs produced by A. dexteroporum, and could be used as early indicators of contamination associated with the ingestion of seafood.

Emerging Marine Biotoxins.

The emergence of marine biotoxins in geographical areas where they have never been reported before is a concern of considerable impact on seafood contamination, and consequently, on public health [...].

Reversible alopecia associated with high blood mercury levels and early menopause: a report of two cases.

OBJECTIVE: The aim of this study was to report on two women in early menopause with alopecia and high mercury (Hg) levels which reversed with a decrease in toxic levels. METHODS: Retrospective chart review and case studies in a reproductive endocrinology practice. RESULTS: A 43-year-old woman initially evaluated for early menopause later experienced sudden circumscribed hair loss on the scalp. Blood tests indicated elevated Hg levels and further investigation revealed a diet high in tuna. Levels fell with elimination of dietary tuna. Another woman, 39 years old was complaining of severe hot flashes, night sweats, and menstrual irregularity also developed alopecia. Treated unsuccessfully for low testosterone, blood tests indicated high Hg levels and simultaneous hair loss was observed; recommendation to alter diet, including fish intake, was followed by a reversal of alopecia, along with a decrease in blood Hg levels. Literature searches were conducted with a focus on Hg toxicity or poisoning with symptom of alopecia. CONCLUSIONS: Women of reproductive age frequently seek treatment for what is thought to be hormone-related hair loss especially at menopause. Two women demonstrated a strong temporal correlation to high Hg levels associated with early menopause, which was reversible. The development of alopecia in the setting of mild Hg intoxication has not been reported in the medical literature despite its appearance in the popular press. Measurement of Hg levels should be considered in women with alopecia and its relationship to early menopause is unclear but bears further research.

Using Precision Environmental Health Principles in Risk Evaluation and Communication of the Deepwater Horizon Oil Spill.

This article provides a description of the rationale and processes adopted by the Gulf Coast Health Alliance: Health Risks related to the Macondo Spill consortium to evaluate and communicate the risk of exposure to polycyclic aromatic hydrocarbons (PAHs) in seafood over several years following the Deepwater Horizon disaster and subsequent oil spill. We examined gaps in knowledge associated with PAH toxicity following exposure to petrogenic (oil-derived) PAHs by studying the metabolic fate of PAHs and their potential toxicity using sophisticated analytical methods. Using the data generated, we developed a risk communication strategy designed to meet the needs of the stakeholder communities including a consumption guideline calculator, a web-based tool to reconcile seafood consumption with risk of adverse health effects.

Variation in the biological half-life of methylmercury in humans: Methods, measurements and meaning.

BACKGROUND: Understanding methylmercury (MeHg) toxicity requires a complete understanding of its fundamental toxicokinetic and toxicodynamic characteristics in the human body. The biological half-life (t1/2) of MeHg is a kinetic property that directly influences the body burden of Hg that results from repeated exposures such as can occur with fish and seafood consumption. The t1/2 of MeHg in humans is approximately 50 days, equivalent to an elimination rate (kel) of 0.014 day-1. However, numerous studies report a wide range of half-life values (t1/2 < 30 to >120 days), demonstrating that significant variation in the biological process of MeHg elimination exists. This variation is a source of considerable uncertainty in deriving a meaningful reference dose for MeHg applicable to all individuals in a population. SCOPE OF REVIEW: First, we summarize fundamentals of MeHg toxicokinetics, emphasizing the central role that biological half-life plays in MeHg dosimetry. We next present important considerations for how kinetic analyses are performed. We provide an example of how MeHg half-life variation directly influences the body burden and, in certain contexts, can result in MeHg levels exceeding the US EPA Reference Dose. We then survey existing studies that report MeHg half-life determinations in people. MAJOR CONCLUSIONS: Recent advances in methods of determining MeHg kinetics in people have made individualized assessment of MeHg elimination rates more accurate and readily obtainable. GENERAL SIGNIFICANCE: Characterization of MeHg half-life, particularly in vulnerable individuals, such as pregnant women and children, will diminish the remaining toxicokinetic uncertainty surrounding MeHg exposures and will better inform the risk assessment process.

Ethnic, Geographic, and Genetic Differences in Arsenic Metabolism at Low Arsenic Exposure: A Preliminary Analysis in the Multi-Ethnic Study of Atherosclerosis (MESA).

We investigated the effect of candidate variants in AS3MT (arsenic (III) methyltransferase) with urinary arsenic metabolites and their principal components in a subset of 264 participants in the Multi-Ethnic Study of Atherosclerosis (MESA). Urinary arsenic species, including inorganic arsenic (iAs), monomethylarsonate (MMA), dimethylarsinate (DMA), and arsenobetaine (Ab), were measured using high performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICPMS) and corrected for organic sources from seafood consumption by regressing Ab on arsenic species using a validated method. Principal components of arsenic metabolism were also used as independent phenotypes. We conducted linear regression of arsenic traits with allelic dosage of candidate single nucleotide polymorphisms (SNPs) rs12768205 (G > A), rs3740394 (A > G), and rs3740393 (G > C) measured using Illumina MetaboChip. Models were stratified by non-Hispanic white vs. all other race/ethnicity and adjusted for age, sex, arsenic exposure, study site, and population stratification. Consistent with previous studies, rs12768205 showed evidence for strongest association (non-Hispanic white: iAs% -0.14 (P 0.83), MMA% -0.66 (0.49), DMA% 0.81(0.49); other race/ethnicity: 0.13 (0.71), -1.21 (0.09), 1.08 (0.20)). No association, however, passed the strict Bonferroni p-value. This was a novel study among an ethnically diverse population exposed to low arsenic levels.

Mercury transport and human exposure from global marine fisheries.

Human activities have increased the global circulation of mercury, a potent neurotoxin. Mercury can be converted into methylmercury, which biomagnifies along aquatic food chains and leads to high exposure in fish-eating populations. Here we quantify temporal trends in the ocean-to-land transport of total mercury and methylmercury from fisheries and we estimate potential human mercury intake through fish consumption in 175 countries. Mercury export from the ocean increased over time as a function of fishing pressure, especially on upper-trophic-level organisms. In 2014, over 13 metric tonnes of mercury were exported from the ocean. Asian countries were important contributors of mercury export in the last decades and the western Pacific Ocean was identified as the main source. Estimates of per capita mercury exposure through fish consumption showed that populations in 38% of the 175 countries assessed, mainly insular and developing nations, were exposed to doses of methylmercury above governmental thresholds. Our study shows temporal trends and spatial patterns of Hg transport by fisheries. Given the high mercury intake through seafood consumption observed in several understudied yet vulnerable coastal communities, we recommend a comprehensive assessment of the health exposure risk of those populations.

Toxicological Investigations on the Sea Urchin Tripneustes gratilla (Toxopneustidae, Echinoid) from Anaho Bay (Nuku Hiva, French Polynesia): Evidence for the Presence of Pacific Ciguatoxins.

The sea urchin Tripneustes gratilla (Toxopneustidae, Echinoids) is a source of protein for many islanders in the Indo-West Pacific. It was previously reported to occasionally cause ciguatera-like poisoning; however, the exact nature of the causative agent was not confirmed. In April and July 2015, ciguatera poisonings were reported following the consumption of T.gratilla in Anaho Bay (Nuku Hiva Island, Marquesas archipelago, French Polynesia). Patient symptomatology was recorded and sea urchin samples were collected from Anaho Bay in July 2015 and November 2016. Toxicity analysis using the neuroblastoma cell-based assay (CBA-N2a) detected the presence of ciguatoxins (CTXs) in T.gratilla samples. Gambierdiscus species were predominant in the benthic assemblages of Anaho Bay, and G.polynesiensis was highly prevalent in in vitro cultures according to qPCR results. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses revealed that P-CTX-3B was the major ciguatoxin congener in toxic sea urchin samples, followed by 51-OH-P-CTX-3C, P-CTX-3C, P-CTX-4A, and P-CTX-4B. Between July 2015 and November 2016, the toxin content in T.gratilla decreased, but was consistently above the safety limit allowed for human consumption. This study provides evidence of CTX bioaccumulation in T.gratilla as a cause of ciguatera-like poisoning associated with a documented symptomatology.

Hair mercury (Hg) levels, fish consumption and semen parameters among men attending a fertility center.

General population exposure to methylmercury (MeHg), the most common organic mercury compound found in the environment, occurs primarily through the consumption of contaminated fish and shellfish. Due to limited studies and lack of consideration of effect modification by fish consumption, it remains uncertain if exposure to mercury affects semen parameters. Thus, we investigated whether hair Hg levels, a biomarker of mercury exposure, were associated with semen parameters among men attending an academic fertility center, and whether this relationship was modified by intake of fish. This analysis included 129 men contributing 243 semen samples who were enrolled in the Environment and Reproductive Health (EARTH) Study between 2005 and 2013, and had data of hair Hg, intake of fish and semen parameters available. Hair Hg levels were assessed using a direct mercury analyzer. Intake of fish was collected using a validated food frequency questionnaire. Semen parameters were analyzed following WHO 2010 evaluation criteria. Generalized linear mixed models with random intercepts accounting for within-man correlations across semen samples were used to evaluate the association of hair Hg levels and semen parameters adjusting for age, BMI, smoking status, abstinence time and alcohol intake. Effect modification by total fish intake (≤1.68 vs. >1.68 servings/week) was tested. The median hair Hg levels of the men was 0.72ppm and ranged from 0.03 to 8.01ppm; almost 30% of the men had hair Hg levels >1ppm. Hair Hg levels were positively related with sperm concentration, total sperm count, and progressive motility, after adjusting for potential confounders and became attenuated after further adjustment for fish intake. Specifically, men in the highest quartile of hair mercury levels had 50%, 46% and 31% higher sperm concentration, total sperm count and progressive motility, respectively, compared to men in the lowest quartile. These associations were stronger among men whose fish intake was above the study population median. Semen volume and normal morphology were unrelated to hair Hg levels. These results confirmed exposure to MeHg through fish intake and showed the important role of diet when exploring the associations between heavy metals and semen parameters among men of couples seeking fertility care. Further research is needed to clarify the complex relationship between fish intake and Hg, and potential effects on male reproductive health, specifically, semen parameters.

Mercury poisoning in a fisherman working on a pelagic fishing vessel due to excessive tuna consumption.

OBJECTIVE: To report the case of a fisherman who developed chronic mercury poisoning due to excessive consumption of tuna while working on a pelagic fishing vessel. CASE REPORT: A 48-year-old male deep-sea fisherman developed paresthesia and pain in both legs while working at sea. He continued working for over 4 months on a pelagic fishing vessel but was eventually unable to function normally as his condition deteriorated. Upon arrival on land, he received specialist treatment, including imaging studies, for 2 months; however, the cause of the symptoms was not identified. An examination of his occupational history revealed that he had worked as a crew member on a pelagic fishing vessel catching tuna for the last 2 years and consumed tuna for two or more meals per day, every day. Two months after discontinuation of tuna consumption, he was tested for mercury. The result showed an elevated blood mercury level (BML) of 21.79 µg/l. Based on the half-life of mercury, the BML was evaluated as 38.70-53.20 µg/l when he was on board. Four months after discontinuing tuna consumption, his BML decreased to 14.18 µg/l, and the symptoms were almost ameliorated. The person responsible for preparing meals on a pelagic fishing ship should be aware that fish may contain high levels of heavy metals and should prepare meals for crew members according to the recommended levels. Crew members should also be aware that fish and shellfish may contain mercury, and hence, they should consume only an appropriate amount.