Differences in microcystin production and genotype composition among Microcystis colonies of different sizes in Lake Taihu

The cyanobacterium Microcystis, which occurs as colonies of different sizes under natural conditions, can produce toxic microcystins (MCs). To monitor the toxicity and assess the risk of Microcystis blooms in Lake Taihu, it is important to investigate the relationship between MC production and Microcystis colony size. In this study, we classified Microcystis collected from Zhushan Bay of Lake Taihu during blooms into four classes with size of <50 μm, 50–100 μm, 100–270 μm and >270 μm and studied their differences in MC production and genetic structure. The results showed that colonies with size of <50, 50–100, 100–270 and >270 μm produced 12.2 ± 11.2%, 19.5 ± 7.9%, 61.3 ± 12.6%, and 7.0 ± 9.6% of total MC, respectively. The proportion of cell density of colonies with size of 50–100, 100–270 and >270 μm was positively correlated with MC concentration during blooms, while that of colonies with size of <50 μm was negatively correlated. The MC cell quota tended to be higher during blooms in colonies with larger size except that of colonies with size of 100–270 μm was higher than that of colonies with size of >270 μm from June 11 to September 16. Colonies with size of <50 μm showed the highest proportion of the less toxic MC congener MC-RR, and colonies with size of >100 μm showed higher proportion of the most toxic MC congener MC-LR than colonies with size of <100 μm. Real-time PCR indicated that larger colonies had higher proportion of potential toxic genotype. Principal component analysis of PCR-denaturing gradient gel electrophoresis profile showed that cpcBA and mcyJ genotype compositions were different between colonies with size of <50 μm and colonies with size of >50 μm, and cpcBA genotype composition was also different among colonies with size of 50–100 μm, 100–270 μm and >270 μm. These results indicated that MC cell quota and congener composition were different in Microcystis colonies with different sizes in Lake Taihu during blooms, and the differences in MC production in colonies with different size resulted chiefly from the difference in their genotype composition. Therefore, the authorities of water quality monitoring and drinking water supply service in Lake Taihu should be alert that the toxicity of Microcystis colony with different size was different during blooms, and the high abundance of colonies larger than 50 μm could be an indicator of relatively high bloom toxicity.     

Analysis of dissolved microcystins in surface water samples from Kovada Lake, Turkey

Dissolved (extracellular) microcystin (MC) concentrations were determined at 3 sampling stations on Lake Kovada, Turkey. The dominant species of cyanobacteria found in August and September of 2006 were Microcystis aeruginosa, Synechococcus sp., Phormidium limosum, Phormidium formosa and Planktothrix limnetica. MC concentrations in water were measured by ELISA and MC variants were examined by HPLC-PDA. Quantitative analysis by HPLC indicated that five MC variants (MC-LR, -RR, -LA, -LW, -LF) were identified in water samples from Kovada Lake. The maximum concentration of dissolved MC-LW was 98.9 µg l^− 1 in October. MC-LR was only detected in May at a concentration of 0.5 µg l^− 1. The cross reactivity of the antibody (MC10E7) to variants such as MC-LA MC-LW & MC-LF was low. Hence the results determined by ELISA were lower than those determined by HPLC in September and October samples due to differences in the specificity of the antibody to MC variants. Total extraceullar MCs was quantified by ELISA and ranged from 0.73 to 48.5 µg MC-LR equivalents l^− 1, which in some cases exceeded the WHO provisional Guideline Value for MC-LR in drinking water. This study confirms that the lakes of Turkey should be monitored for toxic cyanobacteria and for MCs to avoid or reduce the potential exposure of people to these health hazards.     

Simultaneous determination of microcystin contaminations in various vertebrates (fish, turtle, duck and water bird) from a large eutrophic Chinese lake, Lake Taihu, with toxic Microcystis blooms

This is the first to conduct simultaneous determination of microcystin (MC) contaminations in multi-groups of vertebrates (fish, turtle, duck and water bird) from Lake Taihu with Microcystis blooms. MCs (-RR, -YR, -LR) in Microcystis scum was 328 μg g^− 1 DW. MCs reached 235 μg g^− 1 DW in intestinal contents of phytoplanktivorous silver carp, but never exceeded 0.1 μg g^− 1 DW in intestinal contents of other animals. The highest MC content in liver of fish was in Carassius auratus (150 ng g^− 1 DW), followed by silver carp and Culter ilishaeformis, whereas the lowest was in common carp (3 ng g^− 1 DW). In livers of turtle, duck and water bird, MC content ranged from 18 to 30 ng g^− 1 DW. High MC level was found in the gonad, egg yolk and egg white of Nycticorax nycticorax and Anas platyrhynchos, suggesting the potential effect of MCs on water bird and duck embryos. High MC contents were identified for the first time in the spleens of N. nycticorax and A. platyrhynchos (6.850 and 9.462 ng g^− 1 DW, respectively), indicating a different organotropism of MCs in birds. Lakes with deaths of turtles or water birds in the literatures had a considerably higher MC content in both cyanobacteria and wildlife than Lake Taihu, indicating that toxicity of cyanobacteria may determine accumulation level of MCs and consequently fates of aquatic wildlife.     

Dynamics of microcystin production and quantification of potentially toxigenic Microcystis sp. using real-time PCR

Cyanobacterial blooms in eutrophied water body are generally composed of various genotypes with or without microcystin-producing genes (mcy gene cluster). Thus there is a need for quantification of potent toxin producing strains. The present study aimed at identifying microcystin variants and its producer strains in Durgakund pond, Varanasi, India, based on quantification of cpcBA-IGS and mcyA (condensation domain) genes using real-time PCR and LC-MS. Increase in microcystin concentrations was correlated with increase in mcyA copy number and the level of pigments (chlorophyll a, phycocyanin and carotenoids). Also, selected environmental factors (water temperature, light irradiance, rainfall, pH, N and P) and the concentration of microcystin variants (MC-LR, -RR and -YR) were also assessed in samples during May 2010 to April 2011 to establish the possible correlation among these parameters. Nutrients favored cyanobacterial bloom but it could not be correlated with the levels of microcystin variants and seemed to be geographically specific. Microcystis sp. dominant in the pond comprised potentially toxigenic cells. The ratio of potentially toxigenic Microcystis sp. to that of total Microcystis sp. ranged from 0% to 14%. Such studies paved the way to identify and quantify the most potent microcystin producer in the tropical aquatic body.     

Spatial isolation favours the divergence in microcystin net production by Microcystis in Ugandan freshwater lakes

It is generally agreed that the hepatotoxic microcystins (MCs) are the most abundant toxins produced by cyanobacteria in freshwater. In various freshwater lakes in East Africa MC-producing Microcystis has been reported to dominate the phytoplankton, however the regulation of MC production is poorly understood. From May 2007 to April 2008 the Microcystis abundance, the absolute and relative abundance of the mcyB genotype indicative of MC production and the MC concentrations were recorded monthly in five freshwater lakes in Uganda: (1) in a crater lake (Lake Saka), (2) in three shallow lakes (Lake Mburo, George, Edward), (3) in Lake Victoria (Murchison Bay, Napoleon Gulf). During the whole study period Microcystis was abundant or dominated the phytoplankton. In all samples mcyB-containing cells of Microcystis were found and on average comprised 20 ± 2% (SE) of the total population. The proportion of the mcyB genotype differed significantly between the sampling sites, and while the highest mcyB proportions were recorded in Lake Saka (37 ± 3%), the lowest proportion was recorded in Lake George (1.4 ± 0.2%). Consequently Microcystis from Lake George had the lowest MC cell quotas (0.03-1.24 fg MC cell⁻¹) and resulted in the lowest MC concentrations (0-0.5 μg L⁻¹) while Microcystis from Lake Saka consistently showed maximum MC cell quotas (14-144 fg cell⁻¹) and the highest MC concentrations (0.5-10.2 μg L⁻¹). Over the whole study period the average MC content per Microcystis cell depended linearly on the proportion of the mcyB genotype of Microcystis. It is concluded that Microcystis populations differ consistently and independently of the season in mcyB genotype proportion between lakes resulting in population-specific differences in the average MC content per cell.     

Whole lake selective withdrawal experiment to control harmful cyanobacteria in an urban impoundment

Different environmental conditions support optimal growth by Aphanizomenon and Microcystis in Ford Lake, Michigan, USA, based on weekly species biovolume and water chemistry measurements from June through October 2005-2007. Experimental withdrawal of hypolimnetic water through the outlet dam was conducted in 2006, with 2005 and 2007 acting as control years, to test theory regarding management of nuisance and toxic cyanobacteria. The dynamics of Aphanizomenon and Microcystis blooms in Ford Lake appear to be driven largely by NO₃⁻ concentrations, with higher levels shifting the advantage to Microcystis (P < 0.0001). Aphanizomenon was most successful with a mean TN:TP ratio (mol:mol) of 48.3:1, whereas Microcystis thrived with a mean ratio of 70.1:1. Withdrawal of hypolimnetic water successfully destabilized the water column and led to higher levels of NO₃⁻ and the near elimination of the Aphanizomenon bloom in 2006 (P < 0.0001). Selective withdrawal did not reduce Microcystis biovolume or microcystin toxicity. Microcystis biovolume and NO₃⁻ levels were positively correlated with microcystin toxin (P = 0.01) and jointly accounted for 30.5% of the variability in the data. Selective withdrawal may be a viable management option for improving water quality under certain circumstances. To fully address the problem of nuisance and toxic algal blooms in Ford Lake, however, an integrated approach is required that targets cyanobacteria biovolume dynamics as well as conditions suited for toxin production.     

Exposure of the marine deposit feeder Hydrobia ulvae to sediment spiked with LAS congeners

The lethal and sub-lethal toxicity of LAS congeners to the mollusc gastropod Hydrobia ulvae were assessed in spiked sediment bioassays. This complements the little knowledge available to date on mixture effects in the sediment compartment. The LAS homologues joint effect was concentration additive (∑TU_i = 0.8-1). As opposed to the 10-d LC10 based on the sediment associated LAS concentration (91-330 mg/kg) which was independent of the homologue chain length, the LC10 based on the dissolved LAS fraction (0.804-0.068 mg/L) decreased as the homologue chain length increased from 10 to 13 carbons. The quantitative structure-activity relationship (QSAR) derived from these data was log (1/LC10 (mol/L)) = 0.64 log K_ow + 4.40 (n = 5; r² = 0.76; s = 0.24). It showed an apparent higher toxicity compared to the typical QSAR for polar narcosis in water-only systems probably due to the simultaneous exposure of the snail to LAS through the dissolved and the sediment associated fractions. The egestion rate of the surviving snails recovered after few days' exposure (1-d NOEC: 40-107 mg/kg, 9-d NOEC: 65-190 mg/kg) which suggests that the organisms were able to acclimate to LAS during the exposure.     

In-situ identification of iron electrocoagulation speciation and application for natural organic matter (NOM) removal

In this work, iron speciation in electrocoagulation (EC) was studied to determine the impact of operating parameters on natural organic matter (NOM) removal from natural water. Two electrochemical EC parameters, current density (i) and charge loading rate (CLR), were investigated. Variation of these parameters led to a near unity current efficiency (φ = 0.957 ± 0.03), at any combination of i in a range of 1–25 mA/cm² and CLR in a range of 12–300 C/L/min. Higher i and CLR led to a higher bulk pH and limited the amount of dissolved oxygen (DO) reduced at the cathode surface due to mass transfer limitations. A low i (1 mA/cm²) and intermediate CLR (60 C/L/min) resulted in low bulk DO (<2.5 mg/L), where green rust (GR) was identified by in-situ Raman spectroscopy as the primary crystalline electrochemical product. Longer electrolysis times at higher i led to magnetite (Fe₃O₄) formation. Both higher (300 C/L/min) and lower (12 C/L/min) CLR values led to increased DO and/or increased pH, with lepidocrocite (γ-FeOOH) as the only crystalline species observed. The NOM removal of the three identified species was compared, with conditions leading to GR formation showing the greatest dissolved organic carbon removal, and highest removal of the low apparent molecular weight (<550 Da) chromophoric NOM fraction, determined by high performance size exclusion chromatography.     

Impact of toxic cyanobacteria on gastropods and microcystin accumulation in a eutrophic lake (Grand-Lieu, France) with special reference to Physa (= Physella) acuta

Hepatotoxic microcystins (MCs) produced by cyanobacteria are known to accumulate in gastropods following grazing of toxic cyanobacteria and/or absorption of MCs dissolved in water, with adverse effects on life history traits demonstrated in the laboratory. In the field, such effects may vary depending on species, according to their relative sensitivity and ecology. The aims of this study were to i) establish how various intensities of MC-producing cyanobacteria proliferations alter the structure of gastropod community and ii) compare MC tissue concentration in gastropods in the field with those obtained in our previous laboratory experiments on the prosobranch Potamopyrgus antipodarum and the pulmonate Lymnaea stagnalis. We explored these questions through a one-year field study at three stations at Grand-Lieu Lake (France) affected by different intensities of cyanobacteria proliferations. A survey of the community structure and MC content of both cyanobacteria and gastropods was associated with a caging experiment involving P. antipodarum and L. stagnalis. In total, 2592 gastropods belonging to 7 prosobranch and 16 pulmonate species were collected. However, distribution among the stations was unequal with 62% vs 2% of gastropods sampled respectively at the stations with the lowest vs highest concentrations of MC. Irrespective of the station, pulmonates were always more diverse, more abundant and occurred at higher frequencies than prosobranchs. Only the pulmonate Physa acuta occurred at all stations, with abundance and MC tissue concentration (≤ 4.32 µg g DW^− 1) depending on the degrees of MC-producing cyanobacteria proliferations in the stations; therefore, P. acuta is proposed as a potential sentinel species. The caging experiment demonstrated a higher MC accumulation in L. stagnalis (≤ 0.36 µg g DW^− 1 for 71% of individuals) than in P. antipodarum (≤ 0.02 µg g DW^− 1 for 12%), corroborating previous laboratory observations. Results are discussed in terms of differential gastropod sensitivity and MC transfer through the food web.     

黄浦江源水中藻类和微囊藻毒素状况调查

对黄浦江源水中的藻类和微囊藻毒素-LR和-RR的含量进行了检测,结果表明:黄浦江源水中的微囊藻毒素浓度和藻类数目相对较低,藻类数目为(30~700)×104个/L,总MC-LR浓度为100~250 ng/L,总MC-RR浓度为450~650 ng/L;黄浦江源水中的MC-RR浓度比MC-LR浓度高两倍左右,总MC-RR浓度最高为650 ng/L,总MC-LR浓度最高为250 ng/L;受黄浦江水力和环境条件的影响,藻类在河道内大量死亡、裂解导致黄浦江源水中溶解性微囊藻毒素与总微囊藻毒素浓度的比值比一般源水的高很多。     

Seasonal occurrence and toxicity of Microcystis spp. and Oscillatoria tenuis in the Lebna Dam, Tunisia

Physicochemical and biological water quality, including the total microcystin concentrations, was investigated for the first time from January to December 2005 in the Lebna Dam, Tunisia. Microcystin levels and characterization of the different microcystin variants present were measured by protein phosphatase (PP2A) inhibition assays and by LC/MS/MS, respectively. Nutrient values were high, with total inorganic nitrogen and phosphorus concentrations ranging from 0.05 to 8.4 mg L(-1) and from 0.03 to 1.37 mg L(-1), respectively. However, the chlorophyll-a concentrations were very low with a peak (5.32 microg L(-1)) on 20 September 2005 at 9 m depth water samples. Microscopic examination of the phytoplankton samples showed the dominance in the autumn of three morphospecies of the genus Microcystis and the species Oscillatoria tenuis. The total (particulate and dissolved) microcystin concentrations at the surface and at 9 m depth water samples ranged between 0.008 and 1.73, and 0.005 and 5.57 microg microcystin (MC)-LR equivalent L(-1), respectively, with a peak on 20 September. The presence of the microcystin synthetase genes (mcyA, -B, and -C) in the lysates of the three morphospecies of the genus Microcystis and the species O. tenuis indicated that these species were responsible for the microcystin production in this system. The analysis of the field cyanobacterial sample extract containing these species by LC/MS/MS revealed the presence of two microcystin variants: microcystin-LR (MC-LR) and microcystin-YR (MC-YR).     

MC-LR分子印迹聚合物的制备及富集分离性能研究

以微囊藻毒素（MC）-LR作为印迹分子,采用溶胀聚合法制备了分子印迹聚合物,通过红外吸收光谱、热重分析、电镜扫描等技术对聚合物进行了表征,并考察了其对水中微囊藻毒素的富集分离性能。结果表明,当溶液中MC-LR的平均初始浓度为0．86mg／L时,经吸附后平均浓度降为0．07mg／L,聚合物的吸附容量为39．50μmol／g,对其去除率达91％以上。此外,该分子印迹聚合物对MC-LR具有良好的识别能力和特异性吸附能力,这为去除水中的藻毒素提供了一种新的方法。     

Biochemical stress response in tetradifon exposed Daphnia magna and its relationship to individual growth and reproduction

Environmental risk assessment of chemicals toxicity requires the use of costly and labor-intensive chronic data and short-term tests provide additional information. Energy budget is used by the animals for their growth, reproduction, and metabolism and it is reduced in case of toxic stress. Tetradifon acaricide is frequently used in the European Mediterranean region and it is implicated in aquatic environmental pollution. Previous studies showed that the EC₅₀-24 h of tetradifon on Daphnia magna was 8.92 mg/L. Based on that, D. magna were exposed to sublethal tetradifon concentrations of 0.10, 0.18, 0.22 and 0.44 mg/L for five days in order to investigate their effect on intermediate metabolism. Caloric content was determined as biomarker of tetradifon toxicity. Results were analyzed using one-way analysis of variance (ANOVA) and Duncan's significant difference test was used to find differences between groups (α was set at p = 0.05). Daphnids energy content decreased as tetradifon concentration increased. At 120-h caloric content was depleted > 51% at pesticide concentrations of 0.18 mg/L and higher. In order to determine a possible link between the 5-d test and the 21-d chronic test, animals under short-term test were exposed to the same pesticide concentrations known to cause adverse effects on reproduction, growth and survival. Results from the present study indicated a good correlation between the proposed 5-day test and daphnid energy budget. Comparison between both, caloric content results and the chronic effect values obtained using life-table studies, suggested a good fit between them. These studies can be used as earlier, predictive and useful tests with comparable results to the classic chronic ones. Our results indicate that caution must be done about the use of tetradifon in the aquatic environment.     

Short- and long-term dynamics of the toxic potential and genotypic structure in benthic populations of Microcystis

Microcystis colonies are known to overwinter on the surface of the sediment of freshwater ecosystems. However, little is known about the genotypic and toxicological dynamics of Microcystis populations during this benthic life stage. In this study, we report a two-year-long survey of benthic populations of Microcystis, which had spent from a few days to more than six years in the sediment. In order to avoid any interaction with the planktonic proliferations, we chose two deeply buried benthic populations, which could be easily dated. Quantitative PCR on mcyB gene and protein phosphatase inhibition assays were performed to measure their toxic potential, and their genotypic structure was assessed by Capillary Electrophoresis-Single Strand Conformation Polymorphism (CE-SSCP), based on 16S-23S Intergenic Transcribed Spacer (ITS). The microcystin content of the cells seemed to change sharply during the first few months of benthic survival, whereas this content was low and decreased steadily after several years of benthic life. No genetic selection was observed in either the proportion of potentially toxic clones or the ITS sequences for any of the populations considered. From these results, the benthic life stage of Microcystis appears to preserve the structure and the composition of the population over a far larger time scale than classical overwintering period. Finally, some genotypes were common in both of the benthic populations, even though they originated from planktonic blooms that had developed five years apart, suggesting a major overlap of planktonic proliferations in successive years.     

Multi-scale strategies for the monitoring of freshwater cyanobacteria: reducing the sources of uncertainty

Cyanobacterial blooms are a frequent phenomenon in eutrophic freshwaters worldwide and are considered potential hazards to ecosystems and human health. Monitoring strategies based on conventional sampling often fail to cover the marked spatial and temporal variations in cyanobacterial distribution and fluctuating toxin concentrations inherent to cyanobacterial blooms. To deal with these problems, we employed a multi-scale approach for the study of a massive Microcystis bloom in Tajo River (Spain) utilizing 1) remote sensing techniques, 2) conventional water sampling and 3) analysis of chemotypical subpopulations. Tajo River at the study area is influenced by high temperatures waters diverted upstream from a nuclear power plant, the presence of a dam downstream and a high nutrient load, which provide optimal conditions for massive cyanobacterial proliferation. MERIS imagery revealed high Chl-a concentrations that rarely fell below 20 μg L⁻¹ and moderate spatiotemporal variations throughout the study period (March-November 2009). Although the phytoplanktonic community was generally dominated by Microcystis, sampling points highly differed in cyanobacterial abundance and community composition. Microcystin (MC) concentrations were highly heterogeneous, varying up to 3 orders of magnitude among sampling points, exceeding in some cases WHO guideline values for drinking and also for recreational waters. The analysis of single colonies by MALDI-TOF MS revealed differences in the proportion of MC-producing colonies among points. The proportion of toxic colonies showed a highly significant linear correlation with total MC: biovolume ratio (r² = 0.9; p < 0.001), evidencing that the variability in toxin concentrations can be efficiently addressed by simple analysis of subpopulations. We propose implementing a multi-scale monitoring strategy that allows covering the spatiotemporal heterogeneities in both cyanobacterial distribution (remote sensing) and MC concentrations (subpopulation analysis) and thereby reduce the main sources of uncertainty in the assessment of the risks associated to bloom events.     

Effect of cadmium, lead and arsenic on the oviposition, hatching and embryonic survival of Biomphalaria glabrata

Biomphalaria glabrata is a widespread freshwater gastropod mollusc. The easy aquaculture of these organisms allow its use as an accessible tool for contamination bioassays. B. glabrata showed marked metabolic responses when exposed to cadmium, lead and arsenic. Those responses could also affect the reproduction of the snails. Taking into account this hypothesis, B. glabrata were exposed for 96 h (acute laboratory bioassays) to different concentrations of cadmium (0.1, 0.05 and 0 mg/L), lead (0.5, 0.1, 0.05 and 0 mg/L) and arsenic (0.5, 0.1, 0.05 and 0 mg/L). Snails were removed from the aquaria while eggs were left in the same contaminant concentrations. The effect of the assayed toxicants on snail reproduction was registered as the alterations of the total number of laid eggs (TNLE), hatching time and embryonic survival.At 0.10 mg/L cadmium significantly decreased the TNLE (p < 0.05) and no embryos survived. The lowest assayed level (0.05 mg/L) of cadmium, delayed the hatching time twice when it was compared with the control group (p < 0.01).Lead decreased the TNLE at 0.5 mg/L level (p < 0.01). The other assayed doses (0.05 and 0.10 mg/L) also decreased embryonic survival significantly (p < 0.05 and p < 0.01 respectively) and extended twice the time to hatching (p < 0.01). The 0.50 mg/L level killed all embryos.Arsenic at all studied concentrations decreased the TNLE (p < 0.05) while the hatching time was increased by 50%. Embryo survival only decreased at the highest level (0.5 mg/L) of arsenic assayed.In summary, the acute exposure (96 h) to cadmium lead and arsenic, altered the reproduction of B. glabrata, modifying the TNLE, hatching time and embryonic survival.     

Short-term bacterial community composition dynamics in response to accumulation and breakdown of Microcystis blooms

Short-term bacterial community composition (BCC) dynamics in response to accumulation and breakdown of Microcystis blooms were examined by conducting in situ mesocosm experiments with varying levels of Microcystis sp. biomass, ranging from 15 to 3217 μg/L as measured by chlorophyll-a concentration in the freshwater water column. The BCC was assessed by means of terminal restriction fragment length polymorphism (T-RFLP) of 16S ribosomal RNA genes followed by cloning and sequencing of selected samples. The results showed that the composition of both free-living and particle-attached bacterial communities changed during the accumulation and breakdown phases of a Microcystis bloom, and differences were also evident with different levels of Microcystis biomass. The relative abundance of bacteria affiliated with Micrococcineae and Legionellales increased in general after amendment with Microcystis. Significant correlation between the relative abundance of Micrococcineae and breakdown of Microcystis biomass was also observed. Canonical correspondence analysis (CCA) showed that the changes in the free-living and particle-attached bacterial community were mostly related to the changes in the concentrations of chlorophyll-a, dissolved organic carbon (DOC), dissolved oxygen (DO) and pH, which were mainly induced by the breakdown of Microcystis biomass. Overall, our study revealed the following: i) accumulation of Microcystis blooms and their breakdown have strong impacts on bacterial community composition; ii) there might be saprophytic association between Micrococcineae and decomposition of Microcystis biomass; iii) it is necessary to reveal potential associations between Legionellales organisms and Microcystis blooms in eutrophic freshwater lakes.     

Sublethal toxicity and biotransformation of pyrene in Lumbriculus variegatus (Oligochaeta)

The aim of this work was to study the toxicity and biotransformation of polyaromatic hydrocarbon (PAH) pyrene in the oligochaete aquatic worm, Lumbriculus variegatus. PAHs are ubiquitous environmental pollutants that pose a hazard to aquatic organisms, and metabolizing capability is poorly known in the case of many invertebrate species. To study the toxicity and biotransformation of pyrene, the worm was exposed for 15 days to various concentrations of water-borne pyrene. The dorsal blood vessel pulse rate was used as a sublethal endpoint. Pyrene biotransformation by L. variegatus was studied and the critical body residues (CBR) were estimated for pyrene toxicity. The toxicokinetics of pyrene uptake was evaluated. A combination of radiolabeled (¹⁴C) and nonlabeled pyrene was used in the exposures, and liquid scintillation counting (LSC) and high-pressure liquid chromatography were employed in both water and tissue residue analyses. The results showed that L. variegatus was moderately able to metabolize pyrene to 1-hydroxypyrene (1-HP), thus demonstrating that the phase-I-like oxidizing enzyme system metabolizes pyrene in L. variegatus. The amount of the 1-HP was 1-2% of the amount of pyrene in the worm tissues. The exposure to pyrene reduced the blood vessel pulse rate significantly (p < 0.05), showing that pyrene had a narcotic effect. The estimated CBRs remained constant during the exposure time, varying from 0.120 to 0.174 mmol pyrene/kg worm wet weight. The bioconcentration factors (BCF) decreased as exposure concentration increased. It was suggested that the increased toxicity of pyrene accounted for the decrease in BCFs by lowering the activity of the organism.     

Heterotrophic microorganism Rhodotorula mucilaginosa R30 improves tannery sludge bioleaching through elevating dissolved CO2 and extracellular polymeric substances levels in bioleach solution as well as scavenging toxic DOM to Acidithiobacillus species

The introduction of acid-tolerant heterotrophic microorganisms into sludge bioleaching systems has been proven effective in improving sludge bioleaching processes, and such positive effect is mainly attributed to the biodegradation of low molecular weight organic acids or sludge dissolved organic matter (DOM) toxic to Acidithiobacillus species by the heterotrophic microorganisms introduced. Here we report that elevated dissolved CO₂ concentration and resulting extracellular polymeric substances (EPS) in bioleach solution due to the incorporation of heterotrophic microorganisms also play important roles in improving sludge bioleaching. It was found that in tannery sludge bioleaching system coinoculated with Rhodotorula mucilaginosa R30 and Acidithiobacillus species, dissolved CO₂ concentration in bioleach solution can be elevated from 0.23-0.54 mg/L to 0.76-1.01 mg/L compared to the control inoculated only with Acidithiobacillus species. Correspondingly, the distinct degradation of sludge DOM was also observed in this experiment. It was experimentally demonstrated that the accumulation of CO₂ did greatly enhance the growth of Acidithiobacillus thiooxidans and the decrease rate of pH in the medium. In addition, EPS derived from R. mucilaginosa R30 could bind readily Fe³⁺ in bioleach solution with maximum binding capacity (MBC) of 0.82 mg Fe³⁺ by per mg DOC of EPS secreted and the oxidization activity of EPS-bound Fe³⁺ was decreased but not totally inhibited, indicating that the formation of soluble EPS-Fe(III) complexes enhances, to a certain extent, bioleaching efficiency due to maintaining Fe³⁺ level in solution by inhibiting Fe precipitation occurrence.     

Assessing the ecotoxicity of vinyl chloride using green alga P. subcapitata, nematode C. elegans, and the SOS chromotest in a closed system without headspace

The ecotoxicity of vinyl chloride (VC) was evaluated using green alga, nematode, and the SOS chromotest. The green alga and nematode tested were Pseudokirchneriella subcapitata, and Caenorhabditis elegans, respectively. Because of the tendency of VC to escape from an aqueous exposure medium to the air phase, all tests in the present study were performed in a closed system without headspace to minimize the losses of VC. Previous studies on VC toxicity were performed in an open system or closed system with headspace. VC inhibits the growth of P. subcapitata. The 48-h IC50 value for P. subcapitata exposed to VC was calculated to be 5.15 mg/L. The survival of C. elegans was not influenced at concentrations of up to 60 mg/L; however, VC has an adverse effect on the reproduction of C. elegans. In a stress-related gene expression test using C. elegans, a significant and concentration-dependent expression of heat shock protein 16.2 was observed, indicating that VC induces the stress of C. elegans at the genetic level. The results of the SOS chromotest using Escherichia coli PQ37 showed an IF_max value of 1.11, indicating that VC is not genotoxic. The present study demonstrated that VC has an adverse effect on the algal growth and reproductive and genetic levels of C. elegans. A closed system without headspace is an effective method of testing the aquatic toxicity of volatile organic compounds such as VC.