Ecotoxicity of the fluoroquinolone antibiotic delafloxacin to the water flea Simocephalus vetulus and its offspring under the influence of calcium modulation.

Delafloxacin (DFX), one of the latest additions to the fluoroquinolone antibiotics, is gaining heightened recognition in human therapy due to its potential antibacterial efficacy in a wide range of applications. Concerns have arisen regarding its presence in the environment and its potential interactions with multivalent metals, such as calcium (Ca). The present study investigated the trans- and multigenerational effects of environmentally projected concentrations of DFX (100-400 µg DFX L-1) on individual- and population-level responses of parental S. vetulus (F0) and its descendants (F1) under normal (26 mg L-1) and high (78 mg L-1) Ca conditions. Exposure of the F0 generation to DFX under the normal Ca condition resulted in reduced juvenile body length (JBL), increased age-specific survival rate (lx), indicating prolonged developmental time, reduced age-specific fecundity rate (mx), and decreased population growth rate (rm). Under the high Ca condition, JBL, mx, and rm were adversely affected. Transgenerational effects of DFX existed, as F1 individuals exhibited persistent suppressions in at least one endpoint under both Ca conditions even after being transferred to a clear medium. Continuous exposure of the F1 generation to DFX had negative impacts on JBL, mx, and rm under the normal Ca condition, and on JBL and rm under the high Ca condition. However, cumulative effects were not observed, suggesting the potential development of tolerance to DFX in the F1 organisms. These findings suggest that DFX is a harmful compound for the non-target model organism S. vetulus and reveal a potential antagonism between DFX and Ca. Nevertheless, the interaction between other (fluoro)quinolones and Ca remains unclear, necessitating further research to establish this phenomenon more comprehensively, including understanding the interaction mechanism in ecotoxicological contexts.

Ultrasound-Assisted Lipid Extraction from Chlorella sp.: Taguchi Design and Life Cycle Assessment.

The present study delved into the enhancement of essential oil (EO) extraction process from Chlorella sp. through the implementation of ultrasound-assisted extraction. The Taguchi method was instrumental in determining the ideal parameters for the extraction process, which encompassed ultrasonic amplitude, reaction duration, hexane/ethanol (HE/EtOH) ratio, and processing temperature. The empirical findings indicated that optimal EO yield was realized at an ultrasonic amplitude of 80%, a reaction timeframe of 15 min, a HE/EtOH proportion of 3:1, and a temperature setting of 40 °C. These optimal conditions were further substantiated through additional experimentation, resulting in an EO yield of 18.8 ± 0.2%. A fatty acid profile analysis disclosed that the extracted EO predominantly consisted of long-chain fatty acids (C14-C20), with Palmitic, Heptadecanoic, Oleic, and Linoleic acids featuring as the main components. Nevertheless, a high unsaturation rate of 37.9% in the EO could potentially render it vulnerable to oxidative deterioration during storage, consequently affecting the quality of the ensuing biodiesel. A life cycle assessment of the sonication technique utilized for biodiesel production from Chlorella sp. highlighted that lipid extraction was the principal contributor to global warming and ecotoxicity, as per the CML and TRACI methods. Hence, the ultrasound-assisted extraction of EO from Chlorella sp. appears to be a promising and ecologically viable substitute to conventional techniques employed for biodiesel production.

Toxicity of ciprofloxacin and ofloxacin to Moina macrocopa and investigation of p-value adjustments for (eco)toxicological studies.

Ciprofloxacin (CFX) and ofloxacin (OFX) are commonly found as residual contaminants in aquatic environments, posing potential risks to various species. To ensure the safety of aquatic wildlife, it is essential to determine the toxicity of these antibiotics and establish appropriate concentration limits. Additionally, in (eco)toxicological studies, addressing the issue of multiple hypothesis testing through p-value adjustments is crucial for robust decision-making. In this study, we assessed the no observed adverse effect concentration (NOAEC) of CFX and OFX on Moina macrocopa across a concentration range of 0-400 µg L-1. Furthermore, we investigated multiple p-value adjustments to determine the NOAECs. Our analysis yielded consistent results across seven different p-value adjustments, indicating NOAECs of 100 µg CFX L-1 for age at first reproduction and 200 µg CFX L-1 for fertility. For OFX treatment, a NOAEC of 400 µg L-1 was observed for both biomarkers. However, further investigation is required to establish the NOAEC of OFX at higher concentrations with greater certainty. Our findings demonstrate that CFX exhibits higher toxicity compared to OFX, consistent with previous research. Moreover, this study highlights the differential performance of p-value adjustment methods in terms of maintaining statistical power while controlling the multiplicity problem, and their practical applicability. The study emphasizes the low NOAECs for these antibiotics in the zooplanktonic group, highlighting their significant risks to ecological and environmental safety. Additionally, our investigation of p-value adjustment approaches contributes to a deeper understanding of their performance characteristics, enabling (eco)toxicologists to select appropriate methods based on their specific needs and priorities.

Identifying the impact of land use land cover change on streamflow and nitrate load following modeling approach: a case study in the upstream Dong Nai River basin, Vietnam.

Tri An Reservoir is a vital source of water for agriculture, industry, hydropower, and public usage in Southern Vietnam. Due to human activities, water eutrophication has become a serious problem in recent decades. This study investigated for the first time the impact of land use and land cover (LULC) change on streamflow and nitrate load from the upstream Dong Nai River basin, which is the largest watershed of the reservoir. The study utilized several LULC scenarios, including LULC 2000, 2010, and 2020. The SWAT model was applied to model the watershed during the period 1997-2009. Results showed that the hydrological model performed satisfactorily based on the Nash-Sutcliffe efficiency (NSE) coefficient, the root mean square error observations standard deviation ratio (RSR), and the percent bias (PBIAS). The average simulated values of monthly streamflow and nitrate load were 453.7, 450.0, 446.7 m3/s and 17,699.43, 17,869.13, 17,590.81 tonnes for the LULC 2000, 2010, and 2020 scenarios, respectively. There were no significant differences in streamflow and nitrate load at the basin level under the different LULC scenarios. However, when looking at the subbasin level, there were differences in nitrate load among the scenarios. This suggests that the impacts of LULC on nitrate load may be more pronounced at smaller scales. Overall, our finding underscores the importance of modeling techniques in predicting the impacts of LULC change on streamflow and water quality, which can ultimately aid in the sustainable management of water resources.

Seasonal changes in phytoplankton assemblages and environmental variables in highly turbid tropical estuaries of the Mekong River, Vietnam.

The distribution of phytoplankton assemblages in response to physicochemical variables was assessed using Spearman's correlation and canonical correspondence analysis in four highly turbid estuaries of the Mekong River (MKR) system in Ben Tre Province, Vietnam. During two surveys (September 2017 and April 2018) at 20 sampling sites, a total of 162 species were recorded, with a dominance of diatoms (> 50%). Phytoplankton abundance varied from 3.01 × 105 to 11.85 × 105 cells/L, with the highest cell densities found at the BL2 station in the rainy season, mainly attributed to cyanobacteria during the rainy period, whereas diatoms were dominant in the dry season. Chlorophyll-a concentration in this area was low and decreased from the mouth stations. Similarity analysis distinguished the freshwater and oligohaline regions (characterized by lower phytoplankton abundance in the oligohaline areas) from the mesohaline and polyhaline zones (characterized by higher phytoplankton density in the polyhaline sections), based on the salinity gradient, which mostly explained the spatio-temporal distribution of phytoplankton. In which, freshwater and oligohaline sites were dominated by the diatom Coscinodiscus, the euglenoids Lepocinclis, and the green alga Scenedesmus in the rainy season, while mesohaline and polyhaline stations were dominated only by diatom species (Thalassionema and Skeletonema) in the dry season. High salinity and total dissolved solutes, and low nutrients are the major factors affecting phytoplankton growth and lead to low density and low primary biomass in turbid estuarine systems like the MKR estuaries. Our results further emphasize the importance of taking into account of salinity in understanding seasonal changes of phytoplankton communities, which can provide a valuable baseline data for ecological management strategies in tropical estuarine ecosystems.

Physiological response of Simocephalus vetulus to five antibiotics and their mixture under 48-h acute exposure.

Antibiotics, widely known as major environmental xenobiotics, are increasingly being released into ecosystems due to their essential functions in human health and production. During the COVID-19 pandemic waves, antibiotic use increases remarkably in treating bacterial coinfections. Antibiotics were initially expected only to affect prokaryotes, but recent research has shown that they can disturb the biological systems of eukaryotes, especially vulnerable aquatic creatures, through both direct and indirect processes. However, their toxicity to the freshwater cladoceran Simocephalus vetulus, an essential link in the aquatic food web, has never been evaluated. The effects of four fluoroquinolones (ciprofloxacin: CFX, ofloxacin: OFX, gatifloxacin: GFX, delafloxacin: DFX), tetracycline (TET), and a mixture of these medicines (MIX) on S. vetulus thoracic limb rate (TLR) were examined in this study. After S. vetulus was exposed to 20 and 40 mg GFX L-1, 90% and 100% mortality rates were recorded. At 2.5-10 mg L-1, GFX dramatically lowered the TLR of S. vetulus, resulting in a median effective concentration of 9.69 mg L-1. TLRs increased when the organisms were exposed to 10-40 mg L-1 of CFX and 1.25-40 mg L-1 of OFX. However, DFX and TET exposures did not affect TLRs. Exposure to MIX reduced TLR only at 40 mg L-1, suggesting an antagonistic interaction among the five pharmaceuticals. This study demonstrated that S. vetulus physiological responses to antibiotics, even in the same class, are complex and elusive. Beyond a common additive concentration principle, the antagonistic interaction of antibiotic mixture indicates a high level of uncertainty in terms of ecological dangers. We initially introduce S. vetulus to ecotoxicological studies of antibiotics, presenting the species as a low-cost model for physiological investigations of environmental xenobiotics.

Toxicity of di-2-ethylhexyl phthalate and tris (2-butoxyethyl) phosphate to a tropical micro-crustacean (Ceriodaphnia cornuta) is higher in Mekong River water than in standard laboratory medium.

Plasticizers such as di(2-ethylhexyl) phthalate (DEHP) and tris (2-butoxyethyl) phosphate (TBOEP) are manufactured chemicals produced in high volumes. These chemicals are frequently detected in the aquatic environment and cause toxic effects on organisms. In this study, we assessed the chronic impacts of DEHP and TBOEP, respectively, at the concentration of 100 µg L-1 dissolved in the artificial medium (M4/4) and Mekong River water on life history traits of a tropical micro-crustacean, Ceriodaphnia cornuta, for 14 days. DEHP and TBOEP substantially reduced the survival of C. cornuta. In M4/4 medium, both plasticizers strongly enhanced reproduction but did not influence the growth of C. cornuta. Mekong River water, plasticizers-exposed C. cornuta produced less neonates than those in the control. The detrimental impacts of DEHP and TBOEP on the fitness of C. cornuta were much stronger in natural river water than in M4/4. Our results suggest that plasticizers can cause adverse effects on tropical freshwater cladocerans, particularly in natural water. These results are of a deep concern, as national and international regulatory guidelines which are based on ecotoxicological tests using standard media may not fully capture these effects.

Removal of total nitrogen from wastewater by a combination of Chlorella sp. and audible sound.

In developing countries, nitrogen in the traditional market wastewater is a critical environmental problem. In this study, the microalga Chlorella sp., which was isolated from wastewater, was used to remove the total nitrogen (TN) from conventional market wastewater in combination with audible sound (Vietnamese classical music). In addition, effects of sound exposure on removal efficiency at different initial cell densities were analyzed. Results revealed that music sound control demonstrates potential to improve the removal efficiency. TN removal efficiencies of 96%, 69.5%, and 4.3% were observed for treatments with Chlorella sp./audible sound, Chlorella sp., and without Chlorella sp., respectively. The significance of probability value (p-value) (<0.05) on the paired sample t-test confirmed the critical role of audible sound and Chlorella sp. density on the TN removal in screening experiments. The predicted optimal conditions for TN removal were as follows: a Chlorella sp. density of 4%, an audible sound of 52.5 dB, and a cultivation time of 4.6 days. Results based on statistical analysis revealed that the quadratic models for TN removal are significant at a low p-value (<0.05) and a high predicted coefficient of determination (R2 = 0.9452) value. The obtained statistical results also indicated that most of the variables are significant for the abatement of TN from market wastewater using Chlorella sp.

Pesticide production wastewater treatment by Electro-Fenton using Taguchi experimental design.

In this study, the electro-Fenton (EF) method was applied to remove total organic carbon (TOC) from the pesticide production wastewater containing tricyclazole (TC). Statistical Taguchi method was used to optimize the treatment performance. Analysis of variance (ANOVA) indicated that the polynomial regression model fitted experimental data with R2 of 0.969. The optimal conditions for eliminating 75.4% TOC and 93.7% TC were 0.2 mM of Fe2+, 990 mg/L of Na2SO4, 180 min of reaction time at pH 3 with 2.22 mA/cm2 of current density. The removal of TC present in the wastewater followed the first-order reaction kinetic model (R2 = 0.993); while that was the second-order kinetic model in the case of the TOC removal (R2 = 0.903). In addition, the experimental results and theory approaches (density functional theory and natural bond orbital calculations) also showed the C-N bond breaking and nitrate ions cleavage to ammonia. Acute toxicity of the pesticide wastewater after treatment (PWAT) on microcrustaceans showed that the treated wastewater still exhibited high toxicity against D. magna, with LC50 values of 3.84%, 2.68%, 2.05%, and 1.78% at 24 h, 48 h, 72 h, and 96 h, respectively.

Chronic ecotoxicology and statistical investigation of ciprofloxacin and ofloxacin to Daphnia magna under extendedly long-term exposure.

Ciprofloxacin (CFX) and ofloxacin (OFX) are two of the most often used fluoroquinolone antibiotics, and their residues are found in large amounts in various aquatic settings. However, the toxicity tests of CFX using eukaryotic organisms such as Daphnia magna are inadequate, and the test result of OFX is currently unknown. Therefore, the chronic toxicity test for D. magna was performed during 42 days under exposure to CFX and OFX concentrations of 50, 500, and 5000 µg L-1. All exposure conditions did not cause mortality for D. magna. CFX exposure at 500 µg L-1 resulted in an earlier oogenesis date and increased brood size in the second birth. The Poisson-based generalized linear mixed-effects model revealed that the reduction of fertility was statistically significant for the CFX and OFX exposures at 5000 µg L-1. On the other hand, the production of dead eggs as offspring degradation was also found significantly as maternal D. magna exposed to antibiotics at 5000 µg L-1. In addition, following long-term exposure to antibiotics, maternal adaptation to antibiotics was established for offspring deterioration and fertility. However, the OFX exposure showed that the fertility-suppressed effects continued for a longer period than the CFX exposure. Although no rational explanation has yet been given for the more substantial effect of OFX on reducing fertility than CFX, molecular cell biology and symbiotic microbial flora derived from previous studies could explain our ecotoxicological results. This study is the first report for the OFX chronic toxicities on D. magna by comparing it to the toxicity of CFX. Our study contributes to guiding the future impact assessment of fluoroquinolone antibiotic pollution on ecosystems, including the need for new statistical methods in ecotoxicological studies.

Comparing the performance of machine learning algorithms for remote and in situ estimations of chlorophyll-a content: A case study in the Tri An Reservoir, Vietnam.

Chlorophyll-a (Chl-a) is one of the most important indicators of the trophic status of inland waters, and its continued monitoring is essential. Recently, the operated Sentinel-2 MSI satellite offers high spatial resolution images for remote water quality monitoring. In this study, we tested the performance of the three well-known machine learning (ML) (random forest [RF], support vector machine [SVM], and Gaussian process [GP]) and the two novel ML (extreme gradient boost (XGB) and CatBoost [CB]) models for estimation a wide range of Chl-a concentration (10.1-798.7 µg/L) using the Sentinel-2 MSI data and in situ water quality measurement in the Tri An Reservoir (TAR), Vietnam. GP indicated the most reliable model for predicting Chl-a from water quality parameters (R2 = 0.85, root-mean-square error [RMSE] = 56.65 µg/L, Akaike's information criterion [AIC] = 575.10, and Bayesian information criterion [BIC] = 595.24). Regarding input model as water surface reflectance, CB was the superior model for Chl-a retrieval (R2 = 0.84, RMSE = 46.28 µg/L, AIC = 229.18, and BIC = 238.50). Our results indicated that GP and CB are the two best models for the prediction of Chl-a in TAR. Overall, the Sentinel-2 MSI coupled with ML algorithms is a reliable, inexpensive, and accurate instrument for monitoring Chl-a in inland waters. PRACTITIONER POINTS: Machine learning algorithms were used for both remote sensing data and in situ water quality measurements. The performance of five well-known machine learning models was tested Gaussian process was the most reliable model for predicting Chl-a from water quality parameters CatBoost was the best model for Chl-a retrieval from water surface reflectance.

Toxic cyanobacteria and microcystin dynamics in a tropical reservoir: assessing the influence of environmental variables.

Toxic cyanobacterial blooms (TCBs) have become a growing concern worldwide. The present study investigated the dynamic of toxic cyanobacteria and microcystin (MC) concentrations in the Tri An Reservoir (TAR), a tropical system in Vietnam, with quantitative real-time polymerase chain reaction (qPCR) and high-performance liquid chromatography (HPLC), respectively. The results of the qPCR quantification revealed that Microcystis was the dominant group and the primary MC producer in the TAR. Potentially toxigenic cyanobacteria varied from 1.2 × 104 to 1.58 × 107 cells/mL, and the mean proportion of toxic Microcystis to that of the total toxic cyanobacteria varied from 21 to 88%. Microcystin concentrations in raw water and sediment samples often peaked during June to October as blooms occurred and varied from 0.27 to 6.59 µg/L and from 1.79 to 544.9 ng/g in wet weight, respectively. The results of this study indicated that conditions favoring Microcystis proliferation lead to the selection of more toxic genotypes. Water temperature and light availability were not driving factor in the formation of TCBs in the TAR. However, the high loads of total nitrogen (TN), phosphate, and total phosphorus (TP) into the water via rainfall runoff in combination with a high total suspended solid (TSS) and decreased water level during the early months of the rainy seasons did lead to a shift in Microcystis blooms and higher proportions of toxic genotypes of Microcystis in the TAR. This research may provide more insight into the occurrence mechanism of TCBs in tropical waters. The strategy to control TCB problems in tropical regions should be focused on these limnological and hydrological parameters, in addition to a reduction in nitrogen and phosphorus loading.

Estimation of nitrogen and phosphorus concentrations from water quality surrogates using machine learning in the Tri An Reservoir, Vietnam.

Surface water eutrophication due to excessive nutrients has become a major environmental problem around the world in the past few decades. Among these nutrients, nitrogen and phosphorus are two of the most important harmful cyanobacterial bloom (HCB) drivers. A reliable prediction of these parameters, therefore, is necessary for the management of rivers, lakes, and reservoirs. The aim of this study is to test the suitability of the powerful machine learning (ML) algorithm, random forest (RF), to provide information on water quality parameters for the Tri An Reservoir (TAR). Three species of nitrogen and phosphorus, including nitrite (N-NO2-), nitrate (N-NO3-), and phosphate (P-PO43-), were empirically estimated using the field observation dataset (2009-2014) of six surrogates of total suspended solids (TSS), total dissolved solids (TDS), turbidity, electrical conductivity (EC), chemical oxygen demand (COD), and biochemical oxygen demand (BOD5). Field data measurement showed that water quality in the TAR was eutrophic with an up-trend of N-NO3- and P-PO43- during the study period. The RF regression model was reliable for N-NO2-, N-NO3-, and P-PO43- prediction with a high R2 of 0.812-0.844 for the training phase (2009-2012) and 0.888-0.903 for the validation phase (2013-2014). The results of land use and land cover change (LUCC) revealed that deforestation and shifting agriculture in the upper region of the basin were the major factors increasing nutrient loading in the TAR. Among the meteorological parameters, rainfall pattern was found to be one of the most influential factors in eutrophication, followed by average sunshine hour. Our results are expected to provide an advanced assessment tool for predicting nutrient loading and for giving an early warning of HCB in the TAR.

Inland harmful cyanobacterial bloom prediction in the eutrophic Tri An Reservoir using satellite band ratio and machine learning approaches.

In recent years, Tri An, a drinking water reservoir for millions of people in southern Vietnam, has been affected by harmful cyanobacterial blooms (HCBs), raising concerns about public health. It is, therefore, crucial to gain insights into the outbreak mechanism of HCBs and understand the spatiotemporal variations of chlorophyll-a (Chl-a) in this highly turbid and productive water. This study aims to evaluate the predictable performance of both approaches using satellite band ratio and machine learning for Chl-a concentration retrieval-a proxy of HCBs. The monthly water quality samples collected from 2016 to 2018 and 23 cloud free Sentinel-2A/B scenes were used to develop Chl-a retrieval models. For the band ratio approach, a strong linear relationship with in situ Chl-a was found for two-band algorithm of Green-NIR. The band ratio-based model accounts for 72% of variation in Chl-a concentration from 2016 to 2018 datasets with an RMSE of 5.95 µg/L. For the machine learning approach, Gaussian process regression (GPR) yielded superior results for Chl-a prediction from water quality parameters with the values of 0.79 (R2) and 3.06 µg/L (RMSE). Among various climatic parameters, a high correlation (R2 = 0.54) between the monthly total precipitation and Chl-a concentration was found. Our analysis also found nitrogen-rich water and TSS in the rainy season as the driving factors of observed HCBs in the eutrophic Tri An Reservoir (TAR), which offer important solutions to the management of HCBs in the future.

Response of microcystin biosynthesis and its biosynthesis gene cluster transcription in Microcystis aeruginosa on electrochemical oxidation.

Microcystin-LR (MC-LR), which is one of the most commonly found microcystins (MCs) in fresh water, has been proved to be a potential tumour promoter and classified as 2B by the International Agency for Research on Cancer. MC-LR decomposition and inhibition of MC-LR production in Microcystis aeruginosa were investigated under electrolysis condition using an electrolysis cell consisting of Ti/Pt electrodes and Nafion membrane. The relationship between the decrease in MC-LR concentration and transcription of MC-LR synthesis gene clusters was determined by performing real-time reverse transcription polymerase chain reaction (RT-qPCR) to monitor changes in the levels of transcription encoding mcyB and mcyD (cDNA to DNA) in M. aeruginosa NIES 1086 under electrolysis condition and three different conditions (i.e. oxygenated, air aerated and unaerated) as controls. Cell density decreased from day 2 under electrolysis than under the three controls. Intracellular MC-LR concentration was approximately 33 fg cell-1 under electrolysis from days 4 to 8, while those in the other conditions ranged in 40-50 fg cell-1. The mcyB transcription continuously decreased from day 2 to nondetectable level in day 6 under electrolysis, while this transcription was stabilised under the three controls. This result suggested that oxidative stress, such as hydroxyl radicals, played an important role in the down-regulation of mcyB and mcyD gene transcription level and the MC-LR concentration and cell density of M. aeruginosa.

An overview of the accumulation of microcystins in aquatic ecosystems.

Cyanotoxins produced by toxic cyanobacteria pose a major, worldwide environmental threat to freshwater ecosystems. Microcystins (MCs) are considered to be the most hazardous groups. Indeed, some of the largest aquatic ecosystems on the earth are being contaminated with MCs. Questions have arisen regarding their transfer and bioaccumulation in natural environment. This review summarizes the present state of knowledge regarding toxic cyanobacteria and MCs, with a specific focus on their distribution in different components of aquatic ecosystems. Their accumulation in water columns, aquatic animals, plants, and sediments is summarized. MCs have been contaminating all areas of the aquatic ecosystems. Of these, the water column was the most contaminated with MCs and served as an intermediate transmission substation. Via this route, MCs could enter to other stations such as sediment, animals, aquatic and terrestrial plants. Therefore, the use of water contaminated with MCs may induce food chain contaminations with considerable health risks.

First report on free and covalently bound microcystins in fish and bivalves from Vietnam: Assessment of risks to humans.

The free and covalently bound microcystins (MCs) in 3 fish and 2 bivalves from the Dau Tieng Reservoir in Vietnam were investigated for the first time in the present study. The results showed that all species were contaminated with MCs. Our findings indicate that eating the muscle of fish from the Dau Tieng Reservoir is safe but that eating the bivalves is not safe during toxic cyanobacterial bloom episodes. Environ Toxicol Chem 2017;36:2953-2957. © 2017 SETAC.

Microcystin accumulation and biochemical responses in the edible clam Corbiculaleana P. exposed to cyanobacterial crude extract.

We investigated the accumulation and effects of cyanobacterial crude extract (CCE) containing microcystins (MCs) on the edible clam Corbiculaleana P. Toxic effects were evaluated through the activity of antioxidant and detoxification enzymes: catalase (CAT), superoxide dismutase (SOD), and glutathione-S-transferases (GSTs) from gills, foot, mantle and remaining soft tissues. Clams were exposed to CCE containing 400µg MC-LReq/L for 10days and were then kept in toxin-free water for 5days. Clam accumulated MCs (up to 3.41±0.63µg/g dry weight (DW) of unbound MC and 0.31±0.013µg/g DW of covalently bound MC). Detoxification and antioxidant enzymes in different organs responded differently to CCE during the experiment. The activity of SOD, CAT, and GST in the gills and mantle increased in MC-treated clams. In contrast, CAT and GST activity was significantly inhibited in the foot and mostly only slightly changed in the remaining tissues. The responses of biotransformation, antioxidant enzyme activity to CCE and the fast elimination of MCs during depuration help to explain how the clam can survive for long periods (over a week) during the decay of toxic cyanobacterial blooms in nature.

Microcystin uptake and biochemical responses in the freshwater clam Corbicula leana P. exposed to toxic and non-toxic Microcystis aeruginosa: Evidence of tolerance to cyanotoxins.

We investigated the accumulation and adverse effects of toxic and non-toxic Microcystis in the edible clam Corbicula leana. Treated clams were exposed to toxic Microcystis at 100 µg of MC (microcystin)-LReq L-1 for 10 days. The experimental organism was then placed in toxin-free water and fed on non-toxic Microcystis for the following 10 days for depuration. Filtering rates (FRs) by C. leana of toxic and non-toxic Microcystis and of the green alga Chlorella vulgaris as a control were estimated. Adverse effects were evaluated though the activity of catalase (CAT), superoxide dismutase (SOD) and glutathione S-transferase (GST). Clam accumulated MCs (up to 12.7 ± 2.5 µg g-1 dry weight (DW) of free MC and 4.2 ± 0.6 µg g-1 DW of covalently bound MC). Our results suggest that although both toxic and non-toxic cyanobacteria caused adverse effects by inducing the detoxification and antioxidant defense system, the clam was quite resistant to cyanotoxins. The estimated MC concentration in C. leana was far beyond the World Health Organization's (WHO) provisional tolerable daily intake (0.04 µg kg-1 day-1), suggesting that consuming clams harvested during cyanobacterial blooms carries a high health risk.