Bioconcentration and transfer of the organophorous flame retardant 1,3-dichloro-2-propyl phosphate causes thyroid endocrine disruption and developmental neurotoxicity in zebrafish larvae.

Organophosphate flame retardants are emerging environmental contaminants, although knowledge of their health risks is limited. Here, thyroid hormone homeostasis and neuronal development was studied in the progeny of adult zebrafish exposed to tris(1,3-dichloro-2-propyl) phosphate (TDCPP). Adult zebrafish were exposed to TDCPP (0, 4, 20, and 100 µg/L) for 3 months. Increased generation of reactive oxygen species and reduced survival rates was observed in exposed F1 larvae. We also observed a significant decrease in plasma thyroxine and 3,5,3'-triiodothyronine levels in F0 females and F1 eggs/larvae. The mRNA and protein expression of factors associated with neuronal development (e.g., α1-tubulin, myelin basic protein, and synapsin IIa) were significantly downregulated in exposed F1 larvae, as was the level of the neurotransmitters dopamine, serotonin, gamma amino butyric acid, and histamine. Larval locomotion was significantly decreased in exposed fish, but there was no effect on acetylcholinesterase activity. Bioconcentration of TDCPP was observed in F0 fish. TDCPP was also detected in F1 eggs following parental exposure, indicating maternal transfer of this compound. This study uniquely shows that TDCPP can be transferred to the offspring of exposed adults, causing thyroid endocrine disruption and developmental neurotoxicity.

Relationship of proteomic variation and toxin synthesis in the dinoflagellate Alexandrium tamarense CI01 under phosphorus and inorganic nitrogen limitation.

Paralytic shellfish toxins (PSTs) are originated from cyanobacteria and dinoflagellates, including Alexandrium tamarense, the common dinoflagellate species. In this study, a toxic dinoflagellate strain of A. tamarense CI01 was selected for studying the PSTs' concentration and the related protein variation during the whole cell cycle under different nutrient conditions. High-performance liquid chromatography, 2-D DIGE and Western blotting were used collectively for protein profiling and identification. Results showed that the toxin content was suppressed under nitrogen limiting condition, but enhanced in phosphorous limiting medium. Based on the results of proteomics analysis, 7 proteins were discovered to be related to the PSTs biosynthesis of A. tamarense CI01, including S-adenosylhomocysteine hydrolase, ornithine cyclodeaminase, argininosuccinate synthase, methyluridine methyltransferase cystine ABC transporter, phosphoserine phosphatase, argininosuccinate synthase and acyl-CoA dehydrogenase, which corresponds to the metabolism of the methionine, cysteine, ornithine, arginine and proline. Moreover, some photosynthesis relating proteins also increased their expression during PST synthesis period in A. tamarense CI01, such as phosphoenolpyruvate carboxylase, chloroplast phosphoglycerate kinase, peridinin-chlorophyll α-binding protein, Mg(2+) transporter protein and chloroplast phosphoglycerate kinase. The above findings are in support of our hypothesis that these proteins are involved in toxin biosynthesis of A. tamarense CI01, but cause-and-effect mechanisms need to be investigated in further studies.

Comparison of three protein extraction procedures from toxic and non-toxic dinoflagellates for proteomics analysis.

Three methods for extraction and preparation of high-quality proteins from both toxic and non-toxic dinoflagellates for proteomics analysis, including Trizol method, Lysis method and Tris method, were compared with the subsequent protein separation profiles using 2-D differential gel electrophoresis (2-D DIGE), Coomassie Blue and silver staining. These methods showed suitability for proteins with different pIs and molecular weights. Tris method was better for low molecular weight and low pI protein isolation; whereas both Lysis and Trizol method were better for high-molecular weight and high pI protein purification. Trizol method showed good results with Alexandrium species and Gynodinium species, and the background in gel was much clearer than the other two methods. At the same time, only Lysis method caused breaking down of the target proteins. On the other hand, Trizol method obtained higher concentration of ribulose-1,5-bisphosphate carboxylase/oxygenase proteins by Western-blotting, while Tris method was the best for peridinin-chlorophyll-protein complexes protein and T1 protein preparation. DIGE was better than Coomassie Blue and silver staining, except for some limitations, such as the high cost of the dyes, relatively short shelf life and the requirements for extensive and special image capturing equipment. Some proteins related to PSTs synthesis in dinoflagellates are hydrophobic with high molecular weight or binding on membranes and Trizol method performed better than Tris method for these proteins. The Trizol method and 2-D DIGE were effective combination for proteomics investigations of dinoflagellates. This procedure allows reliable and high recovery efficiency of proteins from dinoflagellates for better understanding on their occurrence and toxin-production for physiological and biochemical information.

Distribution and assessment of Pb in the supergene environment of the Huainan Coal Mining Area, Anhui, China.

Coal mining area is highly subject to lead (Pb) pollution from coal mining activities. Several decades of coal mining and processing practices in dozens of coal mines in the Huainan Coal Mining Area (HCMA) have led to the accumulation of massive amounts of coal gangue, which piled in dumps. In order to investigate the impacts of coal gangue dumps on Pb level in the supergene media of the HCMA, a systematic sampling campaign comprising coal gangue, soil, wheat, and earthworm samples was conducted. The average Pb content in the coal mining area soil is 24 mg/kg, which is slightly higher than the associated coal gangues (23 mg/kg) and markedly higher than reference region soil (12.6 mg/kg). Soil in the HCMA present a slight to moderate Pb contamination, which might be related to the weathering and leaching of coal gangue dumps. Lateral distribution of Pb in HCMA soil differed among individual coal mines. The soil profile distribution of Pb depends on both natural and anthropogenic contributions. Average Pb content is higher in roots than in stems, leaves, and wheat husks, while the Pb level in seeds exceeded the maximum Pb allowance for foods (Maximum Levels of Contaminants in Foods of China, GB 2762-2012). Earthworms in the selected area are significantly enriched in Pb, suggesting higher bio-available Pb level in soil in the HCMA.

Spatiotemporal variations of retinoic acids and their metabolites in the marine environment of Hong Kong.

Excessive intake of retinoic acids (RAs) and the oxidative metabolites, 4-oxo-RAs, can lead to abnormal morphological development in animals. This study investigated spatiotemporal variations of concentrations and compositions of these compounds in Hong Kong's seawater and during algal blooms. Total concentrations of the studied compounds in seawater were up to 0.790 and 0.427 ng/L in dry and wet seasons, respectively, though no significant seasonal variation was observed. Spatially, the Deep Bay Water Control Zone was the most enriched area with the studied compounds owing to its semi-enclosed nature and influence from the Pearl River discharge. During algal blooms, the studied compounds were detected up to 4.74 ng/L. Based on calculated risk quotients, the ecological risk of the studied compounds to Hong Kong's marine ecosystems was low. Nevertheless, the occurrence and distribution of these chemicals in the marine environment should be closely monitored where algal blooms frequently occur.

Occurrence of retinoic acids and their metabolites in sewage and their removal efficiencies by chemically enhanced primary treatment and secondary biological treatment.

Sewage treatment plants (STPs) are important in densely populated megacities like Hong Kong to control the release of harmful pollutants from households and industries into the receiving water bodies and maintain water quality for supporting various beneficial uses. This study investigated the occurrence of the teratogenic retinoic acids (RAs) and their oxidative metabolites in sewage and sludge of six selected STPs that treat about 87% of all sewage in Hong Kong annually, and compared the removal efficiencies of these compounds from sewage between two major sewage treatment processes, i.e., chemically enhanced primary treatment (CEPT) and secondary (biological) treatment. The total concentrations of the studied RAs in influent, effluent and sludge from the six selected STPs were found between 21.5 and 33.1 ng/L, 12.0-20.4 ng/L, and 4.33-7.02 ng/g dry weight, respectively. The compounds were dominated by all-trans-RA and 13-cis-RA, together accounting for 46.9-65.6%, 38.4-56.7%, and 62.8-82.8% of the total RAs in influent, effluent and sludge, respectively. The studied RAs could not be satisfactorily removed by both treatment processes with removal efficiencies ranging from 25.4% to 47.4% only, without significant difference in their removal between CEPT and secondary treatment. Based on the calculated hazard quotients of all-trans-RA equivalents (0.248-0.521), the treated effluents from all the six STPs exhibited medium ecological risks to the receiving coastal environment. Therefore, continuous monitoring of these compounds and enhancement of treatment technologies of STPs shall be considered in the future to improve the removal efficiencies of these compounds.

Low-pressure volume retarded osmosis for removal of per- and polyfluoroalkyl substances.

Forward osmosis is an energy efficient process that is capable of recovering high-quality water from secondary wastewater treatment. However, regeneration of the draw solution (DS) is a problem that needs to be addressed. Herein, we developed and optimized a one-step process that does not require additional treatment for the DS. This process, called pressure assisted-volume retarded osmosis (PA-VRO), utilizes naturally occurring pressure with the aid of a small inlet pressure (< 1 bar). Poly(styrenesulfonate) was employed as the DS, for its high solubility in water and large molecular size (∼70,000 Da). Accordingly, real wastewater was employed as the feed solution for 48 h to remove perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) through PA-VRO. The rejection rates for PFOA/PFOS and poly(sodium-4-styrenesulfonate) (PSS) were observed to exceed 98%, after 24 h and 99%, after 48 h. Moreover, there were no traceable amounts of PFOA/PFOS in the DS, and hence the detected concentrations of PFOA and PFOS can be attributed to the residuals from the equipment. Therefore, this well-optimized PA-VRO process can be utilized for potable water production from treated wastewater.

Long-term, selective production of caproate in an anaerobic membrane bioreactor.

Fermentative caproate production from wastewater is attractive but is currently limited by the low product purity and concentration. In this work, continuous, selective production of caproate from acetate and ethanol, the common products of wastewater anaerobic fermentation, was achieved in an anaerobic membrane bioreactor (AnMBR). The reactor was continuously operated for over 522 days without need for chemical cleaning. With an ethanol-to-acetate ratio of 3.0, the effluent caproate concentration was 2.62 g/L on average and the caproate ratio in liquid products reached 74%. Further raising the influent ethanol content slightly increased the effluent caproate level but lowered the product selectivity and resulted in microbial inhibition. The Clostridia (the major caproate-producing bacteria) and Methanobacterium species (which consume hydrogen to alleviate microbial inhibition) was significantly enriched in the acclimated sludge. Our results imply a great potential of utilizing AnMBR to recover caproate from the effluent of wastewater acidogenic fermentation process.

Comparative toxicities of four benzophenone ultraviolet filters to two life stages of two coral species.

The benzophenone (BP) organic ultraviolet (UV) filters have been measured in seawater at ng/L to µg/L levels, but more data on their effects in non-target marine organisms are needed. Corals can be exposed to BPs due to wastewater discharges and coastal recreational activities. In this study, toxicities and bioaccumulation of BP-1 (2,4-dihydroxybenzophenone), BP-3 (oxybenzone), BP-4 (sulisobenzone) and BP-8 (dioxybenzone) to larvae and adults of two coral species, Pocillopora damicornis and Seriatopora caliendrum, were assessed at concentrations ranging from 0.1-1000 µg/L. BP-1 and BP-8 exposure caused significant settlement failure, bleaching and mortality of S. caliendrum larvae [lowest observed effect concentration (LOEC): ≥10 µg/L] compared to the other BPs, while none of the tested compounds and concentrations affected P. damicornis larvae. Nubbins were more sensitive to BP-3, BP-1 and BP-8 than larvae. Overall, BP-1 and BP-8 were more toxic to the two tested species than BP-3 and BP-4, which matches the relative bioaccumulation potential of the four BPs (BP-8 > BP-1 ≈ BP-3 > BP-4). A conservative risk assessment using the effect concentrations derived from this study showed that BP-3, BP-1 and BP-8 pose high or medium risk to the health of corals in popular recreational areas of Taiwan and Hong Kong. Our study suggests that future ecotoxicological studies of corals should take their sensitivities, life stages and metabolic capacities into consideration.

Toxicological effects of two organic ultraviolet filters and a related commercial sunscreen product in adult corals.

Corals are exposed to organic ultraviolet (UV) filters and other personal care product (PCP) ingredients in the environment, but the toxicities of organic UV filters and their related PCP to corals are not well understood. In this study, 7-day exposures were conducted to evaluate the toxicities and bioaccumulation of two organic UV filters, ethylhexylmethoxy-cinnamate (EHMC; octinoxate) and octocrylene (OC) (single- and combined-chemical tests), and diluted sunscreen wash-off water containing both active ingredients to the adult life stage of two hard coral species, Seriatopora caliendrum and Pocillopora damicornis. In the single-chemical tests, death (33.3%) and bleaching (83.3%) were only observed in the 1000 µg/L EHMC treatment of S. caliendrum. In the sunscreen product exposures, 5% sunscreen water (containing 422.34 ±â€¯37.34 µg/L of EHMC and 33.50 ±â€¯7.60 µg/L of OC at Day 0) caused high mortality in S. caliendrum (66.7-83.3%) and P. damicornis (33.3-50%), and tissue concentrations were up to 10 times greater than in the single-chemical exposures; co-exposure to EHMC and OC at similar levels to those in the sunscreen product resulted in bioaccumulation similar to the single-chemical tests. These results confirm the bioaccumulation potential of EHMC and OC and show that other ingredients in sunscreen products may increase the bioavailability of active ingredients to corals and exacerbate the toxicity of sunscreen products. Future studies on the toxicities of PCPs to aquatic organisms should not only focus on the toxicities of active ingredients.

Emissive terbium probe for multiphoton in vitro cell imaging.

A polymeric terbium complex that can be excited by near-infrared excitation at 800 nm via multiphoton absorption processes has been synthesized. This complex has been demonstrated to show strong, observable, three-photon-induced f-f emission in cell imaging. In vitro studies carried out in three carcinoma cell lines (A549, HONE1, and HeLa) have been performed and shown to have low cytotoxicity. This complex is therefore a potential candidate for future infrared excitation imaging dyes.

Pacific Ciguatoxin Induces Excitotoxicity and Neurodegeneration in the Motor Cortex Via Caspase 3 Activation: Implication for Irreversible Motor Deficit.

Consumption of fish containing ciguatera toxins or ciguatoxins (CTXs) causes ciguatera fish poisoning (CFP). In some patients, CFP recurrence occurs even years after exposure related to CTXs accumulation. Pacific CTX-1 (P-CTX-1) is one of the most potent natural substances known that causes predominantly neurological symptoms in patients; however, the underlying pathogenies of CFP remain unknown. Using clinically relevant neurobehavioral tests and electromyography (EMG) to assess effects of P-CTX-1 during the 4 months after exposure, recurrent motor strength deficit occurred in mice exposed to P-CTX-1. We detected irreversible motor strength deficits accompanied by reduced EMG activity, demyelination, and slowing of motor nerve conduction, whereas control unexposed mice fully recovered in 1 month after peripheral nerve injury. Finally, to uncover the mechanism underlying CFP, we detected reduction of spontaneous firing rate of motor cortical neurons even 6 months after exposure and increased number of glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes. Increased numbers of motor cortical neuron apoptosis were detected by dUTP-digoxigenin nick end labeling assay along with activation of caspase 3. Taken together, our study demonstrates that persistence of P-CTX-1 in the nervous system induces irreversible motor deficit that correlates well with excitotoxicity and neurodegeneration detected in the motor cortical neurons.

Acute Exposure to Pacific Ciguatoxin Reduces Electroencephalogram Activity and Disrupts Neurotransmitter Metabolic Pathways in Motor Cortex.

Ciguatera fish poisoning (CFP) is a common human food poisoning caused by consumption of ciguatoxin (CTX)-contaminated fish affecting over 50,000 people worldwide each year. CTXs are classified depending on their origin from the Pacific (P-CTXs), Indian Ocean (I-CTXs), and Caribbean (C-CTXs). P-CTX-1 is the most toxic CTX known and the major source of CFP causing an array of neurological symptoms. Neurological symptoms in some CFP patients last for several months or years; however, the underlying electrophysiological properties of acute exposure to CTXs remain unknown. Here, we used CTX purified from ciguatera fish sourced in the Pacific Ocean (P-CTX-1). Delta and theta electroencephalography (EEG) activity was reduced remarkably in 2 h and returned to normal in 6 h after a single exposure. However, second exposure to P-CTX-1 induced not only a further reduction in EEG activities but also a 2-week delay in returning to baseline EEG values. Ciguatoxicity was detected in the brain hours after the first and second exposure by mouse neuroblastoma assay. The spontaneous firing rate of single motor cortex neuron was reduced significantly measured by single-unit recording with high spatial resolution. Expression profile study of neurotransmitters using targeted profiling approach based on liquid chromatography-tandem mass spectrometry revealed an imbalance between excitatory and inhibitory neurotransmitters in the motor cortex. Our study provides a possible link between the brain oscillations and neurotransmitter release after acute exposure to P-CTX-1. Identification of EEG signatures and major metabolic pathways affected by P-CTX-1 provides new insight into potential biomarker development and therapeutic interventions.

Occurrence and distribution of polybrominated diphenyl ethers (PBDEs) in the dissolved and suspended phases of the sea-surface microlayer and seawater in Hong Kong, China.

This study reports the first data on the concentration and distribution of polybrominated diphenyl ethers (PBDEs) in the sea-surface microlayer (SML), and their enrichment relative to bulk seawater, for coastal waters in the marine environment of Hong Kong, China. Samples were collected in March 2005 at five sample locations and analysed for eight congeners of primary interest, i.e. BDE 28, 47, 99, 100, 153, 156, 183 and 209. Concentration ranges of summation operatorPBDE in the dissolved phase (DP, defined as sum of truly dissolved and colloidal phase) and suspended particulate matter (SPM) of seawater were 31.1-118.7 pg/l (mean 70.7 pg/l), and 25.7-32.5 pg/l (mean 28.1 pg/l), respectively. Concentrations in the SML were generally higher by factor of 1.3-3.6 in the DP (concentration range from 40.2 to 228.2 pg/l, mean 149.2 pg/l) and by 0.3-2.1 in the SPM (concentration range 8.1-69.1 pg/l, mean 38.2 pg/l). Concentrations of PBDEs were general low and below detection limits in samples of an oceanic character and highest in the sheltered waters of Victoria Harbour. The congeners BDE 28, 47, 100 and 183 were most abundant, where BDE 209 was detected only in trace amounts. It is suggested that Hong Kong's marine waters show relatively low levels of PBDE contamination, and these compounds may originate from the disposal of electronic waste in southern China, as well as untreated discharge of wastewater locally.

Perfluorinated compounds in streams of the Shihwa Industrial Zone and Lake Shihwa, South Korea.

Concentrations of perfluorinated alkyl compounds (PFAs), including perfluorooctane sulfonate (PFOS), perfluorohexanesulfonate, perfluorobutanesulfonate, perfluorooctanesulfonamide, perfluorodecanoate, perfluorononanoic acid, perfluorooctanoate (PFOA), perfluoroheptanoate, and perfluorohexanoate, were measured in the streams of the Shihwa and Banweol industrial areas on the west coast of South Korea as well as the adjacent Lake Shihwa (an artificial lake) and Gyeonggi Bay. Perfluorinated alkyl compounds were concentrated from water using solid-phase extraction and were identified and quantified by liquid chromatography/ triple-quadrapole tandem mass spectrometry. Of the PFAs measured, PFOS and PFOA occurred at the greatest concentrations. Concentrations of PFOS ranged from 2.24 to 651 ng/L, and concentrations of PFOA ranged from 0.9 to 62 ng/L. The concentrations of PFOS observed in Lake Shihwa were among the greatest ever measured in the environment. These results suggest local industrial sources of PFOS and PFOA as well as other PFAs. Because of dilution, the greatest concentrations occur in a rather restricted area, near the points of discharge of the streams that empty into the lake. The greatest measured concentration of PFOS exceeded the threshold for effects predicted for predatory birds consuming aquatic organisms continuously exposed to this level.

Distribution of organochlorines in the dissolved and suspended phase of the sea-surface microlayer and seawater in Hong Kong, China.

Polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) were measured in the dissolved and suspended particulate phase in the sea-surface microlayer (SML) and subsurface water (SSW) collected from Hong Kong's coastal environment. The concentration ranges (pg/L) for summation sigmaHCHs, summation sigmaDDTs and summation sigmaPCBs in the SSW dissolved phase (DP i.e. sum of truly dissolved and colloidal phase) were 409-940 (mean 602), 774-5583 (mean 1908) and 266-433 (mean 278), respectively. The concentration ranges (pg/L) for summation sigmaHCHs, summation sigmaDDTs and summation sigmaPCBs in SSW suspended particulate matter (SPM) were <5-85 (mean 59), 358-1369 (mean 787) and 85.6-273 (mean 172), respectively. The enrichment factor of PCBs and OCPs in the SML varied between 1.1 and 4.5 for the DP, and 0.4-8.2 for the SPM. The distribution of contaminants between DP and SPM in both the SML and SSW indicates that particulate matter plays an important role in the distribution and fate of DDTs and PCBs, but not for HCH isomers. The Pearl River Estuary is likely to be a major source of contaminants transported to Hong Kong.

Competitive sorption of heavy metals by water hyacinth roots.

Heavy metal pollution is a global issue severely constraining aquaculture practices, not only deteriorating the aquatic environment but also threatening the aquaculture production. One promising solution is adopting aquaponics systems where a synergy can be established between aquaculture and aquatic plants for metal sorption, but the interactions of multiple metals in such aquatic plants are poorly understood. In this study, we investigated the absorption behaviors of Cu(II) and Cd(II) in water by water hyacinth roots in both single- and binary-metal systems. Cu(II) and Cd(II) were individually removed by water hyacinth roots at high efficiency, accompanied with release of protons and cations such as Ca2+ and Mg2+. However, in a binary-metal arrangement, the Cd(II) sorption was significantly inhibited by Cu(II), and the higher sorption affinity of Cu(II) accounted for its competitive sorption advantage. Ionic exchange was identified as a predominant mechanism of the metal sorption by water hyacinth roots, and the amine and oxygen-containing groups are the main binding sites accounting for metal sorption via chelation or coordination. This study highlights the interactive impacts of different metals during their sorption by water hyacinth roots and elucidates the underlying mechanism of metal competitive sorption, which may provide useful implications for optimization of phytoremediation system and development of more sustainable aquaculture industry.

Ciguatoxin reduces regenerative capacity of axotomized peripheral neurons and delays functional recovery in pre-exposed mice after peripheral nerve injury.

Ciguatera fish poisoning (CFP) results from consumption of tropical reef fish containing ciguatoxins (CTXs). Pacific (P)-CTX-1 is among the most potent known CTXs and the predominant source of CFP in the endemic region responsible for the majority of neurological symptoms in patients. Chronic and persistent neurological symptoms occur in some CFP patients, which often result in incomplete functional recovery for years. However, the direct effects of exposure to CTXs remain largely unknown. In present study, we exposed mice to CTX purified from ciguatera fish sourced from the Pacific region. P-CTX-1 was detected in peripheral nerves within hours and persisted for two months after exposure. P-CTX-1 inhibited axonal regrowth from axotomized peripheral neurons in culture. P-CTX-1 exposure reduced motor function in mice within the first two weeks of exposure before returning to baseline levels. These pre-exposed animals exhibited delayed sensory and motor functional recovery, and irreversible motor deficits after peripheral nerve injury in which formation of functional synapses was impaired. These findings are consistent with reduced muscle function, as assessed by electromyography recordings. Our study provides strong evidence that the persistence of P-CTX-1 in peripheral nerves reduces the intrinsic growth capacity of peripheral neurons, resulting in delayed functional recovery after injury.

Effects of 4-methylbenzylidene camphor (4-MBC) on neuronal and muscular development in zebrafish (Danio rerio) embryos.

The negative effects of overexposure to ultraviolet (UV) radiation in humans, including sunburn and light-induced cellular injury, are of increasing public concern. 4-Methylbenzylidene camphor (4-MBC), an organic chemical UV filter, is an active ingredient in sunscreen products. To date, little information is available about its neurotoxicity during early vertebrate development. Zebrafish embryos were exposed to various concentrations of 4-MBC in embryo medium for 3 days. In this study, a high concentration of 4-MBC, which is not being expected at the current environmental concentrations in the environment, was used for the purpose of phenotypic screening. Embryos exposed to 15 µM of 4-MBC displayed abnormal axial curvature and exhibited impaired motility. Exposure effects were found to be greatest during the segmentation period, when somite formation and innervation occur. Immunostaining of the muscle and axon markers F59, znp1, and zn5 revealed that 4-MBC exposure leads to a disorganized pattern of slow muscle fibers and axon pathfinding errors during the innervation of both primary and secondary motor neurons. Our results also showed reduction in AChE activity upon 4-MBC exposure both in vivo in the embryos (15 µM) and in vitro in mammalian Neuro-2A cells (0.1 µM), providing a possible mechanism for 4-MBC-induced muscular and neuronal defects. Taken together, our results have shown that 4-MBC is a teratogen and influences muscular and neuronal development, which may result in developmental defects.

Identification of a new Irgarol-1051 related s-triazine species in coastal waters.

A previously unknown s-triazine species present in commercially available Irgarol-1051, a booster biocide additive in copper-based antifouling paints for the replacement of organotin-based antifoulants, has been identified in the coastal aquatic environment. After careful isolation, purification and characterization by high resolution MS-MS and (1)H NMR, the molecular structure of that unknown species is found to be N,N'-di-tert-butyl-6-methylthiol-s-triazine-2,4-diamine (designated as M3). Levels of Irgarol-1051, its major degradation product (M1) and the newly identified M3 in the coastal waters of Hong Kong, one of the world's busiest ports located in the southern coast of China, were monitored by SPME-GC-MS and SPME-GC-FID. Water samples from five locations within Hong Kong waters were analysed and the levels of Irgarol-1051, M1 and M3 were found to be 0.1-1.6 microg l(-1), 36.8-259.0 microg l(-1) and 0.03-0.39 microg l(-1), respectively. Our results indicate that M3 is relatively stable against photo- and bio-degradation and may pose considerable risk to primary producer communities in the coastal marine environment.