Assessing the microcystins concentration through optimized protein phosphatase inhibition assay in environmental samples.

Protein phosphatase (PPase) inhibition assay (PPIA) is widely used to analyze the concentration of microcystins (MCs) because it is comparatively less expensive and faster than other assays. This study aimed to optimize the PPIA by determining a suitable reaction terminator and an optimal methanol concentration in the sample. The most suitable reaction time was 90 min, with the corresponding methanol concentration in the sample being 15% or less.

Quantification of toxigenic Microcystis spp. in freshwaters by quantitative real-time PCR based on the microcystin synthetase A gene.

A method to estimate the abundance of toxigenic Microcystis in environmental samples by using quantitative real-time PCR was developed and optimized. The basis of this method is the amplification of a highly conserved region of the mcyA gene within the microcystin synthetase gene cluster. Using this method, the average copy number of mcyA gene per cell in toxigenic Microcystis strains was estimated.

Simultaneous quantification of cyanobacteria and Microcystis spp. using real-time PCR.

In order to develop a protocol to quantify cyanobacteria and Microcystis simultaneously, the primers and probe were designed from the conserved regions of 16S rRNA gene sequences of cyanobacteria and Microcystis, respectively. Probe match analysis of the Ribosomal Database Project showed that the primers matched with over 97% of cyanobacterial 16S rRNA genes, indicating these can be used to amplify cyanobacteria specifically. The TaqMan probe, which is located between two primers, matched with 98.

Establishment of an axenic culture of microcystin-producing Microcystis aeruginosa isolated from a Korean reservoir.

In order to establish an axenic (bacteria-free) culture of Microcystis aeruginosa NIER 10039 isolated from a Korean reservoir, the culture was subjected to sequential treatment, including ultrasonication, washing, and addition of antibiotics. Three broad-spectrum antibiotics, namely, kanamycin, ampicillin, and imipenem, were applied separately in that order. Axenicity of the culture was confirmed by cultivation on bacterial media and observation under epifluorescence and scanning electron microscopes.

Dechlorination of individual congeners in aroclor 1248 as enhanced by chlorobenzoates, chlorophenols, and chlorobenzenes.

Previous investigations showed that three classes of haloaromatic compounds (HACs; chlorobenzoates, chlorophenols, and chlorobenzenes) enhanced the reductive dechlorination of Aroclor 1248, judging from the overall extent of reduction in Cl atoms on the biphenyl. In the present study, we further investigated the kind of polychlorinated biphenyl (PCB) congeners involved in the enhanced dechlorination by four isomers belonging to each class (2,3-, 2,5-, 2,3,5-, and 2,4,6-chlorobenzoates; 2,3-, 3,4-, 2,5-, and 2,3,6-chlorophenols; and 1,2-, 1,2,3-, 1,2,4-, and penta-chlorobenzenes). Although the PCB congeners involved in the enhanced dechlorination varied with the HACs, the enhancement primarily involved paradechlorination of the same congeners (2,3,4'-, 2,3,4,2'- plus 2,3,6,4'-, 2,5,3',4'- plus 2,4,5,2',6'-, and 2,3,6,2',4'- chlorobiphenyls), regardless of the HACs.

Reductive dechlorination of low concentration polychlorinated biphenyls as affected by a rhamnolipid biosurfactant.

We investigated whether the threshold concentration for polychlorinated biphenyl (PCB) dechlorination may be lower in biosurfactant-amended sediments compared with biosurfactant-free samples. At PCB concentrations of 40, 60, and 120 ppm, the surfactant amendment enhanced the PCB dechlorination rate at all concentrations and the rate was also faster at higher concentrations. On a congener group basis, dechlorination proceeded largely with group A (congeners with low threshold) in both surfactant-free and -amended sediments, accumulating mainly group C (residual products of dechlorination) congeners, and surfactant enhanced the dechlorination rate of group A congeners.

Molecular characterization of polychlorinated biphenyl-dechlorinating populations in contaminated sediments.

Polychlorinated biphenyl (PCB)-dechlorinating microorganisms were characterized in PCB-contaminated sediments using amplified ribosomal DNA restriction analysis (ARDRA). The sediments were prepared by spiking Aroclor 1248 into PCB-free sediments, and were inoculated with microorganisms eluted from St. Lawrence River sediments.

Effects of sulfate concentration on the anaerobic dechlorination of polychlorinated biphenyls in estuarine sediments.

In order to determine the effects of sulfate concentration on the anaerobic dechlorination of polychlorinated biphenyls, sediments spiked with Aroclor 1242 were made into slurries using media which had various sulfate concentrations ranging from 3 to 23 mM. The time course of dechlorination clearly demonstrated that dechlorination was inhibited at high concentration of sulfate due to less dechlorination of meta-substituted congeners. When the dechlorination patterns were analyzed by the calculation of Euclidean distance, the dechlorination pathway in the 3 mM sulfate samples was found to be different from that observed in the 13 mM samples, although the extent of dechlorination in these two samples was similar.

Effects of a rhamnolipid biosurfactant on the reductive dechlorination of polychlorinated biphenyls by St. Lawrence River (North America) microorganisms.

The effect of a rhamnolipid biosurfactant on the reductive dechlorination of polychlorinated biphenyls was investigated with the use of clean sediments spiked with Aroclor 1248. The surfactant was added to the contaminated sediments at four different concentrations (5, 10, 25, and 50 microg/g sediment [ppm] on a sediment dry wt basis), and dechlorination was followed over a 40-week period. The rate of overall dechlorination was enhanced at the three highest concentrations.

Bioconcentration and redeposition of polychlorinated biphenyls by zebra mussels (Dreissena polymorpha) in the Hudson River.

The potential impact of zebra mussel infestation on the dynamics of polychlorinated biphenyls (PCBs) in the Hudson River was determined by investigating the biodeposition and bioconcentration of PCBs, using algal food contaminated with 2,5,2'- and 2,4,2',4'-chlorobiphenyls (CBPs) in the laboratory. Approximately 46-90% of the total food was ingested depending on the supply rate. The highest proportion of ingested congeners was found in biodeposits (64+/-11% for 2,5,2'-CBP, and 52+/-6% for 2,4,2',4'-CBP), followed by tissues (17+/-3% for 2,5,2'-CBP, and 23+/-5% for 2,4,2',4'-CBP), and the lowest in shells.

Reductive dechlorination of polychlorinated biphenyls: threshold concentration and dechlorination kinetics of individual congeners in Aroclor 1248.

Reductive dechlorination of individual PCB congeners in Aroclor 1248 was investigated using sediment microorganisms from the St. Lawrence River (NY). No dechlorination was observed at Aroclor concentrations below 40 ppm [137 nmol (g of sediment)(-1)].

Enhancement of microbial PCB dechlorination by chlorobenzoates, chlorophenols and chlorobenzenes.

Abstract We investigated the effects of chlorobenzoates (3-, 2,3-, 2,4-, 2,5-, 2,3,5- and 2,4,6-chlorobenzoate), chlorophenols (2,3-, 3,4-, 2,5-, 2,3,6- and penta-chlorophenol), and chlorobenzenes (1,2-, 1,2,3-, 1,2,4- and penta-chlorobenzene) on polychlorinated biphenyl (PCB) dechlorination and on the enrichment of PCB-dechlorinating microorganisms. When the natural microbial populations eluted from St. Lawrence River sediments were enriched with each of the 15 haloaromatic compounds (HACs) in PCB-free sediments, PCB-dechlorinating microorganisms were found in all but pentachlorophenol-amended sediments.

Kinetics of polychlorinated biphenyl dechlorination by Hudson River, New York, USA, sediment microorganisms.

The kinetics of polychlorinated biphenyl (PCB) dechlorination by Hudson River (New York, USA) sediment microorganisms were investigated using Aroclor 1242 at 10 concentrations ranging from 0 to 900 ppm (0-11.2 micromol Cl/g sediment). The time course of PCB dechlorination and population growth were determined by congener-specific analysis and the most-probable-number technique, respectively, over a 44-week incubation period.