• Journal of the Korean Chemical Society
• /
• v.38 no.10
• /
• pp.741-748
• /
• 1994

A new HPLC method for the analysis of cyanobacterial toxins, i.e. microcystin was developed using cyano-type prepacked cartridge while the conventional method was to utilize ODS cartridge. The cartridge was washed with 0.5 M acetic acid, then microcystins RR and LR were eluted from the cartridge with 30% acetonitrile. A better degree of quantitation was observed than with a ODS cartridge. Especially, in the case of microcystin LR a great difference in peak area was observed.

• Applied Chemistry for Engineering
• /
• v.8 no.4
• /
• pp.610-616
• /
• 1997

Hepatotoxic cyanobacteria, Microcystis species, were collected from the Nakdong River and we could isolate hepatotoxins, microcystin-LR and microcystin-RR, which are also strong inhibitors of protein phosphatase 1 and 2A. From the microcystins, several microcystin derivatives were synthesized and tested on the mouse toxicity in order to establish the structure-activity relationship. Esterification od carboxyl groups of Glu and MeAsp residue produced nontoxic compounds. However, when we reduced the Mdha residue with sodium borohydride into Ala residue, toxicity was still maintained. Also, the change of guanidyl moiety of Arg residue in microcystin-LR into dimethylpyrimidyl moiety did not change the toxicity of microcystins as well. Thus the carboxyl groups seem to play important roles in binding with protein phosphatase 1 and 2A, whereas Mdha residue and the guanidyl moiety of Arg residue do not.

• Journal of the Korean Chemical Society
• /
• v.42 no.1
• /
• pp.51-56
• /
• 1998

It is very difficult to separate and purify the microcystins, cyanobacterial toxins since it exist in a trace level in natural lakes. In this paper, we developed a new analytical method to separate and purify the microcystin RR and LR from the freeze-dried cyanobacterial cells in natural lakes. We used 7.5 g silica gel as a stationary phase and ethyl acetate: isopropanol: water (30: 45: 25) as a mobile phase and microcystins were eluted using an open column. The eluting solvent was collected in a small bottle at the intervals of 3 mL and the fractions were chromatographed with HPLC to confirm the microcystin RR and LR.

• Toxicological Research
• /
• v.22 no.3
• /
• pp.301-306
• /
• 2006

Microcystin-LR(MC-LR), a cyanobacterial toxin produced by Microcystis aeruginosa, causes severe hepatotoxicity. Here we investigated the morphologic changes of rat hepatocyte spheroid induced by exposure of MC-LR($10^{-6}M$) in vitro. In addition, to determine the effects of such toxin in the process of hepatocyte spheroid formation, primarily isolated hepatocytes were incubated with MC-LR and the process of spheroid formation was observed. In both hepatocyte spheroid and suspension culture systems, the morphologic changes caused by MC-LR were noticible at 5 min post exposure and were characterized by the loss of microvilli, cytoplasmic vacuolation, the accumulation of lipid droplets, and blob formation. Especially, the size and numbers of blob on the cell surface were increased as the incubation time prolonged and the appearance of electron dense bodies were observed in the cytoplasm of hepatocyte at 20 min post exposure. Furthermore, bile canaliculi-like structures in the hepatocyte spheroids were slightly widened and the process of spheroids formation was inhibited in the isolated hepatocytes incubated with MC-LR. These results indicate that morphologic changes in. the hepatocyte membrane and organelles seem to be typical events in showing the MC-LR induced hepatotoxic effects and the spheroid culture method might be a useful experimental tool to evaluate hepatoxicity since it reflects the in vivo status of hepatocytes.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.2
• /
• pp.212-218
• /
• 2006

Three out of fourteen Microcystis-dominant cyanobacterial blooms in Central India were found to be toxic to mice ($LD_{50}$ ranging from 35-450 mg bloom dry mass/kg body weight). The liver architecture of the treated mice showed characteristic symptoms of hepatotoxicity relative to the untreated controls, with increased enzyme activities of serum lactate dehydrogenase (LDH), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), and serum glutamate pyruvate transaminase (SGPT). RP-HPLC revealed the presence of microcystin-LR, microcystin-RR, and desmethyl microcystin-RR in the given region to maximum amounts of 390, 1,030, and $860{\mu}g/g$ bloom dry weight, respectively, corresponding to a maximum of 2.8 mg/l microcystin-LR in the lake water. Further confirmation of the microcystin variants was conducted using a MALDI-TOF MS analysis.

• Korean Journal of Ecology and Environment
• /
• v.34 no.3 s.95
• /
• pp.175-184
• /
• 2001

The temporal and diel changes of cyanobacterial cell density, species composition, and cyanobacterial toxins (microcystin-RR, -YR, -LR) were examined for the outflow water of Lake Suwa in Japan from May to October, 1998. The highest total cell densities of Microcystis were observed in July and September, when the dominant phytoplankton was Microcystis ichthyoblabe and M. viridis, respectively. Both the species composition and total cell density of Microcystis affected the variation of the concentration of three microcystin variants. Only microcystin-RR(MC-RR) and -LR (MC-LR) were detected in July when Microcystis ichthyoblabe dominated, while microcystin-RR, -YR (MC-YR) and -LR were detected in August and October when Microcystis viridis dominated. The microcystin concentration and the cell density of Microcystis in the outflow water showed diel variations; the ratio of maximum to minimum value was $3{\sim}20$ fold far microcystin concentration, and $5{\sim}31$ fold for cell density. The diel variations of toxin concentration as well as Microcystis cell density was closely related to the diel variation of wind. During the windy period, when higher speeds occurred in the afternoon hours than morning hours, both the cell density of Microcystis and microcystin concentration tended to increase in the morning and decrease in the afternoon. The results of this study suggest that controlling the timing of lake discharge at the floodgate or intake tower can be useful for water resource management with respect to decreasing cyanobacteria biomass within intake water.

• Korean Journal of Ecology and Environment
• /
• v.38 no.3 s.113
• /
• pp.313-321
• /
• 2005

Based on the result that biological control agent (BCA) increased the dissolved microcystin-LR in a field experiment to control the cyanobacterial bloom (Kim etal., 2005), a laboratory experiment was used to evaluate the effects of dissolved microcystin-LR (MCLR) with different concentrations on abundance, dominance, diversity of phytoplankton community, concentration of chlorophyll a and microcystin concentration in replicated microcosms. The treatments in this laboratory experiment comprised different concentrations of T1 (natural MCLR concentration), T10 (ten times to natural MCLR concentration), and T100 (one hundred times to natural MCLR concentration). MCLR treatment of exclusively Stephanodiscus hantzschii-dominated community in Chonho bridge hardly changed in algal species, but abundance. In Kildong pond, Aulacoseira and Dinobryonrich community was replaced by green algae Scenedesmus-rich community especially in T100 experiment. However, in Yangsoori-Ryukgakji Pond having the highest concentration of initial MCLR, Microcystis aeruginosa was decreased in abundance. Therefore, the treatment of BCA to control M. aeruginosa severely changed the Phytoplankton community in term of algal species, abundance (chlorophyll a) and dissolved microcystin-LR via a high release of MCLR.

• Proceedings of the Korea Environmental Mutagen Society Conference
• /
• 2002.11a
• /
• pp.148-148
• /
• 2002

• Korean Journal of Ecology and Environment
• /
• v.47 no.spc
• /
• pp.19-30
• /
• 2014

Microcystins (MCs) produced by cyanobacteria are severe hepatotoxins for mammalian and protein phosphatase inhibitors. Irrigation water for grain and vegetables is often contaminated with cyanobacteria and microcystin during warm seasons. We assessed the effects of various concentrations (0, 0.01 to $10{\mu}gmL^{-1}$) of microcystin-LR (MC-LR) and microcystin-RR (MC-RR) exposure on Oryza sativa (rice) and Brassica oleraces var. italica (broccoli). The $EC_{50}$ of leaves and roots of rice was 0.9 and $1.1{\mu}gMC-LRmL^{-1}$, respectively. The no observed effect level (NOEL) of rice was less than $0.1{\mu}gmL^{-1}$ ($100{\mu}gL^{-1}$). The $EC_{50}$ of the stems and roots of broccoli was 8.7 and $7.2{\mu}gMC-RRmL^{-1}$, respectively. There was no difference in the germination rate of broccoli among microcystin-RR concentrations. After exposure to 0, 0.01 to $10{\mu}gmL^{-1}$ MC-RR for seven days, 14, 89 and 154 ng mg-1 (dry weight) MC-RR accumulated in B. oleracea. These $EC_{50}$ values showed that microcystin-LR and -RR affected the growth of rice and broccoli. These findings suggest that MC is carried into terrestrial ecosystems via irrigation, and that the biota of higher ecological niches can be influenced by MC through bioaccumulation. Therefore, a guideline for MC concentrations in irrigation water should be set using the NOEL.

• Bulletin of the Korean Chemical Society
• /
• v.26 no.12
• /
• pp.2089-2092
• /
• 2005