Spatial and temporal variations of microcystins in hepatopancreas of a freshwater snail from Lake Taihu

In this paper, spatial and temporal variations of three common microcystins (MC-RR, MC-YR, and MC-LR) in the hepatopancreas of a freshwater snail (Bellamya aeruginosa) were studied monthly in two bays of Lake Taihu. Microcystins (MCs) concentration in hepatopancreas was quantified by liquid chromatography-mass spectrometry (LC-MS). The MCs concentrations in hepatopancreas were higher at Site 1 than those at other sites, which was in agreement with the changes of intracellular MCs concentrations in the water column. There was a significant correlation between MCs concentrations in the hepatopancreas and that in the seston, suggesting that spatial variances of MCs; concentrations in hepatopancreas among the five sites were due to spatial changes of toxic Microcystis cells in the water column. PCCA indicates that in addition to Microcystis, other factors (e.g., water temperature) also substantially affected the accumulation of MCs in hepatopancreas of the snail. (C) 2008 Published by Elsevier Inc.

Bacterial diversity in activated sludge from a consecutively aerated submerged membrane bioreactor treating domestic wastewater

The bacterial diversity of activated sludge from submerged membrane bioreactor (SMBR) was investigated. A 16S rDNA clone library was generated, and 150 clones were screened using restriction fragment length polymorphism (RFLP). Of the screened clones, almost full-length 16S rDNA sequences of 64 clones were sequenced. Phylogenetic tree was constructed with a database containing clone sequences from this study and bacterial rDNA sequences from NCB1 for identification purposes. The 90.6% of the clones were affiliated with the two phyla Bacteroidetes (50%) and Proteobacteria (40%), and beta-, -gamma-, and delta-Proteobacteria accounted for 7.8%, 28.1%, and 4.7%, respectively. Minor portions were affiliated with the Actinobacteria and Firmicutes (both 3.1%). Only 6 out of 64 16S rDNA sequences exhibited similarities of more than 97% to classified bacterial species, which indicated that a substantial fraction of the clone sequences were derived from unknown taxa. Rarefaction analysis of operational taxonomic units (orrUs) clusters demonstrated that 150 clones screened were still insufficient to describe the whole bacterial diversity. Measurement of water quality parameter demonstrated that performance of the SMBR maintained high level, and the SMBR system remained stable during this study.

Experimental studies on the effects of toxic Microcystis aeruginosa PCC7820 on the survival and reproduction of two freshwater rotifers Brachionus calyciflorus and Brachionus rubens

Blooms of Microcystis aeruginosa frequently occur in many eutrophic lakes in China, however, there is very little experimental study on the relationship between Microcystis and rotifers from Chinese waters. The effects of different concentrations of toxic M. aeruginosa PCC7820 on two common freshwater rotifers Brachionus calyciflorus and B. rubens were investigated in laboratory experiments. B. calyciflorus was able to utilize this strain of M. aeruginosa as a food source. However, M. aeruginosa suppressed the survival and reproduction of B. calyciflorus at the highest concentration (10(6) cells/ml) probably due to the inadequate nutrition. B. rubens was inhibited by toxic M. aeruginosa PCC7820 and the inhibition increased with the increasing Microcystis concentration. Our study indicates that the two rotifers have different sensitivities to toxic M. aeruginosa and that toxic cyanobacteria may affect zooplankton community structure by differentially inhibiting the different zooplankton taxa.

The accumulation of alkylphenols in submersed plants in spring in urban lake, China

The concentrations of alkylphenols (APs) were investigated in water, sediments and submersed macrophytes from the Moon Lake, Wuhan city, China. The water samples contained APs, ranging up to 26.4 mu g l(-1) for nonylphenol (NP) and 0.68 mu g l(-1) for octylphenol (OP). APs were found in the sediment samples with concentrations ranging from 4.08 to 14.8 for NP and from 0.22 to 1.25 mu g l(-1) dry weight for OP. The samples from the site near former sewage inlet showed the highest concentrations of APs in both water and sediments. The results of distribution pattern and dynamics of NP and OP in submersed macrophytes of the Moon Lake showed that the two pollutants were all found in Myriophyllum verticillatum, Elodea nuttallii, Ceratophyllum oryzetorum, and Potamageton crispus collected from the Moon Lake. For NP, M. verticillatum had the highest capacity of accumulation, followed by E. nuttallii, C. oryzetorum and P. crispus. However the distribution pattern of OP differed from that of NP, and the highest amount of accumulation was observed in E. nuttallii, followed by M. verticillatum, P. crispus, and C. oryzetorum. The temporal pattern of APs was also observed in submersed macrophytes from March to May, and the highest accumulation period was in May. (c) 2008 Elsevier Ltd. All rights reserved.

Non-microcystin producing Microcystis wesenbergii (Komarek) Komarek (Cyanobacteria) representing a main waterbloom-forming species in Chinese waters

It is well known that several morphospecies of Microcystis, such as Microcystis aeruginosa (Kutzing) Lemmermann and Microcystis viridis (A. Brown) Lemmermann can produce hepatotoxic microcystins. However, previous studies gave contradictory conclusions about microcystin production of Microcystis wesenbergii (Komarek) Komarek. In the present study, ten Microcystis morphospecies were identified in waterblooms of seven Chinese waterbodies, and Microcystis wesenbergii was shown as the dominant species in these waters. More than 250 single colonies of M. wesenbergii were chosen, under morphological identification, to examine whether M. wesenbergii produce hepatotoxic microcystin by using multiplex PCR for molecular detection of a region (mcyA) of microcystin synthesis genes, and chemical analyses of microcystin content by ELISA and HPLC for 21 isolated strains of M. wesenbergii from these waters were also performed. Both molecular and chemical methods demonstrated that M. wesenbergii from Chinese waters did not produce microcystin. (C) 2007 Elsevier Ltd. All rights reserved.

Growth and antioxidant system of Escherichia coli in response to microcystin-RR

Microcystins are a kind of cyclic hepatoxins produced by many species of cyanobacteria. The toxic effects of microcystins on animals and plants have been well studied. However, the reports about the effects of microcystins on microbial cells are very limited. In present paper, Escherichia coli was undertaken to determine the effect of microcystin-RR. These results suggested that microcystin-RR could prolong the growth of E. coli when exposed to high concentrations of microcystin-RR and cause the accumulation of ROS and induce the oxidant stress for a short time. The antioxidant system protects E. coli from oxidative damage.

Kinetic study of the 2-methyl-3-methoxy-4-phenylbutanoic acid produced by oxidation of microcystin in aqueous solutions

Microcystins (MCs) are a family of related cyclic hepatotoxic heptapeptides, of which more than 70 types have been identified. The chemically unique nature of the C20 beta-amino acid, (2S, 3S, 8S, 9S)-3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca4,6-dienoic acid (Adda), portion of the MCs has been exploited to develop a strategy to analyze the entirety. Oxidation of MCs causes the cleavage of MC Adda to form 2-methyl-3-methoxy-4-phenylbutanoic acid (MMPB). In the present study, we investigated the kinetics of MMPB produced by oxidation of the most-often-studied MC variant, MC-LR (L = leucine, R = arginine), with permanganate-periodate. This investigation allowed insight regarding the influence of the reaction conditions (concentration of the reactants, temperature, and pH) on the conversion rate. The results indicated that the reaction was second order overall and first order with respect to both permanganate and MC-LR. The second-order rate constant ranged from 0.66 to 1.35 M/s at temperatures from 10 to 30 degrees C, and the activation energy was 24.44 kJ/mol. The rates of MMPB production can be accelerated through increasing reaction temperature and oxidant concentration, and sufficient periodate is necessary for the formation of MMPB. The initial reaction rate under alkaline and neutral conditions is higher than that under acidic conditions, but the former decreases faster than the latter except under weakly acidic conditions. These results provided new insight concerning selection of the permanganate-periodate concentration, pH, and temperature needed for the oxidation of MCs with a high and stable yield of MMPB.

How polluted is the Yangtze River? Water quality downstream from the Three Gorges Dam

The concentrations of major anions and cations, nitrogen and phosphorus, dissolved and particulate trace elements, and organic pollutants were determined for the middle and lower reaches of the Yangtze River (Changjiang) from below the Three Gorges Dam (TGD) to the mouth at Shanghai in November 2006. The concentration of dissolved inorganic phosphate (DIP) was constant at a low level of 6-8 mu gP/L, but the concentration of nitrate (NO3-) approximately doubled downstream and was closely correlated with K+. This translated to a daily load of well over 1000 It of dissolved inorganic nitrogen (DIN) at Datong. The average concentrations of dissolved Pb (0.078 +/- 0.023 mu g/L), Cd (0.024 +/- 0.009 mu g/L), Cr(0.57 +/- 0.09 mu g/L), Cu (1.9 +/- 0.7 mu g/L), and Ni (0.50 +/- 0.49 mu g/L) were comparable with those in other major world rivers, while As (3.3 +/- 1.3 mu g/L) and Zn (1.5 +/- 0.6 mu g/L) were higher by factors of 5.5 and 2.5, respectively. The trace element contents of suspended particles of As (31 +/- 28 mu g/g), Pb (83 +/- 34 mu g/g), and Ni (52 +/- 16 mu g/g) were close to maximum concentrations recommended for rivers by the European Community (EC). The average concentrations of Cd (2.6 +/- 1.6 mu g/g), Cr (185 +/- 102 mu g/g), Cu (115 +/- 106 mu g/g), and Zn (500 +/- 300 mu g/g) exceeded the EC standards by a factor of two, and Hg (4.4 +/- 4.7 mu g/g) by a factor of 4 to 5. Locally occurring peak concentrations exceed these values up to fourfold, among them the notorious elements As, Hg, and Tl. All dissolved and particulate trace element concentrations were higher than estimates made twenty years ago [Zhang, J., Geochemistry of trace metals from Chinese river/estuary systems: an overview. Estuar Coast Shelf Sci 1995; 41: 631-658.]. The enormous loads of anthropogenic pollutants disposed to the river were diluted by the large water discharge of the Yangtze even during the lowest flow resulting in the relatively low concentration levels of trace elements and organic pollutants observed. We estimated loads of e.g. As, Pb and Ni to the East China Sea to be about 4600 kg As d(-1), 3000 kg Pb d(-1), and 2000 kg Ni d(-1). About 6000 t d(-1) of dissolved organic carbon (DOC) was delivered into the sea at the time of our cruise. We tested for 236 organic pollutants, and only the most infamous were found to be barely above detection limits. We estimated that the load of chlorinated compounds, aromatic hydrocarbons, phenols, and PAHs were between 500 and 3500 kg d(-1). We also detected eight herbicides entering the estuary with loads of 5-350 kg d(-1). The pollutant load, even when at low concentrations, are considerable and pose an increasing threat to the health of the East China Sea ecosystem. (c) 2008 Elsevier B.V. All rights reserved.

Detection and quantitative analysis of polychlorinated biphenyls in tilapia from Hawaiian waters

The purpose of this study was to investigate polychlorinated biphenyls (PCBs) contamination in tilapia (Oreochromis mossambicus) collected from the Manna stream and Ala Wai Canal of O'ahu, an island of the geographically isolated Hawaiian archipelago. Our results show that the average concentrations of PCBs varied from 51.90 to 89.42 ng g(-1) lipid weight for the sampling sites. Relative toxic potencies (RTPs) and toxic equivalencies (TEQs) were determined to be 20.38-40.60 ng TCDD g(-1) lipid weight and 2.89-4.17 ng TEQ g(-1) lipid weight by 7-ethoxy-resorufin-O-deethylase (EROD) activity analysis and calculation of PCB concentrations based on toxic equivalency factors (TEFs), respectively. Penta-chlorinated congeners were found to be predominant, which revealed that Aroclor 1254 was a possible major source of PCBs in our fish samples. PCB 118, an indicator PCBs, constituted more than 55% and 30% of the total PCBs and TEQs, respectively. In addition, PCB 118 was found to have a linear correlation to the total PCBs (R = 0.975) and TEQs (R = 0.782). Detection of concentrated PCBs in Hawaiian waters suggests a potentially adverse impact of this pollutant on human health, as well as ecological systems, and suggests the necessity of environmental monitoring and hazard assessment of PCBs within the Hawaiian Islands. (c) 2008 Published by Elsevier Ltd.

Response of macrozoobenthos communities to ecological engineering remediation in a hypertrophic urban lake

The relationship of macrozoobenthos communities with some environmental variables, and their response to the ongoing restoration measures were studied in a small hypertrophic urban lake near the Yangzte River, China. Twenty taxa including 9 oligochaetes, 7 insects, 2 mollusks and two other animals were found during March 2005 to May 2006. The reappearance of some indigenous macrozoobenthos species showed that the ecological engineering remediation carried out was helpful for the recovery of the macrozoobenthos communities. Through canonical correspondence analysis (CCA), it was detected that temperature (T), conductivity (COND), Secchi depth/deep (SD/Deep), TN, total suspended solids (SS) and chemical oxygen demand (CODcr) were significant environmental factors that influenced the pattern of macrozoobenthos. Limnodrilus hoffmeisteri, Tanypus sp. and Alocinma longicornis could be used as potential bio-indicators in monitoring the development of ecological restoration.

Accumulation of hepatotoxic microcystins in freshwater mussels, aquatic insect larvae and oligochaetes in a large, shallow eutrophic lake (Lake Chaohu) of subtropical China

This paper studied the seasonal changes of two common microcystins (MCs), MC-RR and -LR, in the commercially important mussel Corbicula fluminea in Lake Chaohu, where there occurred dense cyanobacteria. Occasional measurements were also made for MC in the mussel Arconaia lanceolat, the oligochaete Limnodilus hoffineisteri and the insect larva Chironomus sp. Mean MC of C. fluminea was much higher in hepatopancreas than in intestine and foot. Our study is the first to report accumulation of MCs in oligochaetes and aquatic insect larvae. The hi-h contents of MCs in the insect larvae suggest a great possibility for the transfer of MCs to benthos-feeding omnivores like common carp. According to the provisional standard by the WHO, 28.6% of the collected C. fluminea were harmful for human consumption, assuming a daily consumption of 300 by a person. It is recommended that edible mussels should not be collected for human consumption during toxic cyanobacterial blooms in Lake Chaohu.

Effects of Cd2+ and Pb2+ on the substrate biofilms in the integrated vertical-flow constructed wetland

The effects of single Cd2+ and Pb2+, and combined Cd2+ and Pb2+ on dehydrogenase activity and polysaccharide content of the substrate biofilms in the integrated vertical-flow constructed wetland (IVCW) were studied. Dehydrogenase activities decreased linearly with the increasing concentrations of Cd2+ and Pb2+ at different times (6, 24, 72, and 120 h). The activities at both 6 and 24 h were significantly higher than that at 72 and 120 h in the case of single and combined treatments. The single Cd2+ and Pb2+ treatments significantly inhibited dehydrogenase activities at concentrations in excess of 20 mu mol/L Cd2+ and 80 mu mol/L Pb2+, respectively. The inhibitory effect of Cd2+ was much greater than that of Pb2+. At the same time, the combined treatment of Cd2+ and Pb2+ Significantly inhibited dehydrogenase activities at all five concentrations studied and the lowest combined concentration was 1.25 mu mol/L Cd2+ and 5 mu mol/L Pb2+. A synergistic effect of Cd2+ and Pb2+ was observed. On the other hand, polysaccharide contents varied unpredictably with the increasing concentrations of Cd2+ and Pb2+ and extended experimental time. There were no significant statistical differences within the range of concentration and time studied, whether singly or in combination. These results implied that the effects of heavy metals on biofilms should be a concern for the operation and maintenance of constructed wetlands.

Tissue distribution and depuration of the extracted hepatotoxic cyanotoxin microcystins in crucian carp (Carassius carassius) intraperitoneally injected at a sublethal dose

An acute toxicity experiment was conducted by intraperitoneal injection with a sublethal dose of extracted microcystins (MCs), 50 mu g MC-LR (where L = leucine and R = arginine) equivalent/kg body weight (BW), to examine tissue distribution and depuration of MCs in crucian carp (Carassius carassius). Liver to body weight ratio increased at 3, 12, 24, and 48 h postinjection compared with that at 0 h (p < 0.05). MC concentrations in various tissues and aquaria water were analyzed at 1, 3, 12, 24, 48, and 168 h postinjection using liquid chromatography coupled with mass spectrometry (LC-MS). The highest concentration of MCs (MC-RR + MC-LR) was found in blood, 2 -270 ng/g dry weight (DW), followed by heart (3 -100 ng/g DW) and kidney (13 -88 ng/g DW). MC levels were relatively low in liver, gonad, intestine, spleen, and brain. MC contents in gills, gallbladder, and muscle were below the limit of detection. Significant negative correlation was present between MC-RR concentration in blood and that in kidney, confirming that blood was important in the transportation of MC-RR to kidney for excretion. Rapid accumulation and slow degradation of MCs were observed in gonad, liver, intestine, spleen, and brain. Only 0.07% of injected MCs were detected in liver. The recovery of MCs in liver of crucian carp seemed to be dose dependent.

Effects of nutrients on the substrate biofilms in the integrated vertical-flow constructed wetland

A L9 orthogonal array design involving 3 factors (C6H12O6, KNO3 and NaH2PO4) and 3 levels for each (C6H12O6: 0.2, 0.4 or 0.8 g/L; KNO3: 0.4, 0.8 or 1.6 g/L, NaH2PO4: 0.05, 0.1 or 0.2 g/L), was used to study the effects of nutrients on dehydrogenase activity and polysaccharide content of substrate biofilms in the integrated vertical-flow constructed wetland (IVCW). Results showed that C6H12O6 and KNO3 were the main factors for dehydrogenase activity and polysaccharide content of biofilms, respectively. The combinations of three nutrients at different concentrations had different effects on dehydrogenase activity and polysaccharide content of biofilms. The optimal combination for dehydrogenase activity was obtained by locating the concentrations Of C6H12O6, KNO3 and NaH2PO4 at 0.2, 0.8 and 0.05 g, and the optimal combination for polysaccharide content was obtained by locating the concentrations Of C6H12O6, KNO3 and NaH2PO4 at 0.2, 0.4 and 0.2 g/L, respectively. The corresponding maximum activity and polysaccharide content were 5.40 mu g TF/g substrate/12 h and 3454.6 mu g/g substrate, respectively. These results would provide the laboratory foundation for optimizing the purification function of the wetland systems.

Nitrogen dynamics in large shallow eutrophic Lake Chaohu, China

Temporal and spatial dynamics of nitrogen in lake and interstitial water were studied monthly in a large shallow, eutrophic lake in subtropical China from October 2002 to September 2003. The distribution of nitrogen was consistent with the idea that high nitrogen concentrations in the western part of the lake resulted from high levels of the nutrients from the surrounding cities through sewage-drainage systems. Nitrate was the predominant form of nitrogen in the overlying water, while ammonium was predominant in the interstitial water, indicating that strong oxidative nutrient regeneration occurred near the sediment-water interface. Nitrate could be an important dissolved inorganic matter source for phytoplankton, which in turn influenced the seasonal variations of nitrate concentrations in lake water. Significant positive correlation between ammonium fluxes and water temperature was observed and could probably be attributed to the intensified ammonification and nitrate reduction with increased temperature. Positive correlation between ammonium fluxes and algae biomass and Chl a concentrations may indicate that phytoplankton was an important factor driving ammonium fluxes in our study lake, and vice versa that higher fluxes of ammonium supported a higher biomass of the phytoplankton.