Targeting Microcystin (MC), which is most abundantly detected in the North-Han River water area, we analyzed the relationship between the MC biosynthesis gene (mcyA gene), cyanobacteria cell density, and MC concentration, derived an RNA-MC conversion formula, and derived the cyanobacteria. The concentration of MC present in cells was predicted. In the North-Han River waters, the mcyA gene was found mainly at downstream sites of the North-Han River after Muk-Hyeon Stream junction, and higher copy numbers were found on average than other sites. In the Uiam Lake waters upstream of the North-Han River, the mcyA gene copy number increased at the Kong-Ji Stream point, and after September, the mcyA gene copy number decreased throughout the North-Han River waters. The expression of the mcyA gene was concentrated in the short period of summer due to the spatio-temporal difference between upstream and downstream water bodies. The mcyA gene expression level was not only highly correlated with MC concentration, but also correlated with the cell density of Microcystis aeruginosa and Dolichospermum circinale, which are known to biosynthesize MC. Six conversion formulas derived based on the RNA-MC relationship showed statistical significance (p<0.05) and exhibited high correlation coefficients (r) of 0.9 or higher. The expression level of MC biosynthesis gene present in eRNA determines the synthesis of cyanotoxin substances in water, quickly quantifies gene activity, and can be fully utilized for early warning of MC development.

Cyanobacteria (blue green algae) blooms formed in natural water resources due to the environmental pollution produce toxic compounds as secondary metabolites, causing health hazards to both humans and other living beings. Microcystin is a well-known toxin produced by cynobacteria. The present study was undertaken to evaluate varying concentrations and exposure times of two different forms of microcystin, viz., -LR (MCLR) and -LA (MCLA), on the motility and seizure-like behavior of planarian (Dugesia japonica). Compared to control, reduced motility was observed in both the MCLR or MCLA treated groups, but did not differ significantly with increasing concentrations of microcystin. However, the number of seizure-like behaviors were increased dose-dependently in planarian exposed to MLCR or MCLA. Exposure time to microcystine also affected the motility and seizure-like behaviors of planarians; 24 hrs incubation with MCLR, and 48 and 96 hrs exposure to MCLA, showed significantly (p<0.05) lower motility, as compared to the control. Assessing regeneration of the planarians revealed the simultaneous completion of eye formation at day 9 in planarians incubated in the absence or presence of MCLR or MCLA, thereby indicating that exposure to microcystin has no effect on the process. In conclusion, we determined that exposure to microcystins resulted in decrease in the number of motility, and induced abnormal behavior pattern in planarians. Further studies are required to identify the toxicity of microcystin that affects aquatic ecosystems.

NSF developed NSF p477, which covers POU active media systems. Exact types and quantities of cyanotoxins vary. In 2015 USEPA set recommended health advisories for microcystins in drinking water at or below 0.3 μg/L for children less than 6 years old, and at or below 1.6 μg/L for older children and adults. NSF developed and characterized a natural source for microcystin compounds to be used as a contaminant challenge for the POU active media systems being tested. Analytical methodologies were refined to reliably identify and quantify microcystin at levels in the sub part per billion range. Laboratory column studies using various activated carbons were conducted to determine the relative retention of various microcystin congeners, to assess the appropriateness of various congeners for testing purposes.

For several decades, lactic acid bacterium (Lactobacillus graminis: LAB) has been generally recognized as safe. To develop the pan-environmental bio-control agent, algicidal activity of the live LAB cell and its culture filtrate (CF) was examined against Microcystis aeruginosa. LAB cells perfectly lysed M. aeruginosa within 3 days, while the CF had a less effect than the live cells, approximately 78% inhibition of algal growth during a same culture period. The concentration of microcystin in alone culture of M. aeruginosa was 7.1 μg L-1, but gradually increased and leach 158.5 μg L-1 on 10 days. However, LAB cells clearly decreased the microcystin by 10.3 μg L-1 in the same period, approximately 93.5%. CF of LAB showed a strong algicidal activity over 75% between pH 2-7, 91.3% by the treatment of proteinase K, 87.8% by below 3 kDa in particle size, and 75.3% by heat treatment, respectively. Of five solvents, fractions of CF passed through solvents diethyl ether and ethyl acetate showed an obvious algicidal activity in the algal-lawn test. Among 5 fractions purified by silica-gel TLC plate, two spots showed a most strong removal activity on M. aeruginosa. Another analysis of GC indicate that CF contained six representative fatty acids. Even though most of these substance have been known as an anti-algal substance against M. aeruginosa, oleic acid is the most effective. These results suggested that the culture filtrate or specific substances, like a fatty acids, in comparison with live L. graminis can be a successful and eco-friendly agent to control Microcystis bloom.

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 μg mL-1) of microcystin-LR (MC-LR) and microcystin-RR (MC-RR) exposure on Oryza sativa (rice) and Brassica oleraces var. italica (broccoli). The EC50 of leaves and roots of rice was 0.9 and 1.1 μg MC-LRmL-1, respectively. The no observed effect level (NOEL) of rice was less than 0.1 μg mL-1 (100 μg L-1). The EC50 of the stems and roots of broccoli was 8.7 and 7.2 μg MC-RR mL-1, respectively. There was no difference in the germination rate of broccoli among microcystin-RR concentrations. After exposure to 0, 0.01 to 10 μg mL-1 MC-RR for seven days, 14, 89 and 154 ng mg-1 (dry weight) MC-RR accumulated in B. oleracea. These EC50 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.

환경시료에서 마이크로시스틴 분해능을 나타낸 균주 1종을 분리하였고 16S rRNA gene sequence 분석 결과, Microbacterium sp.로 동정되어 Microbacterium sp. MA21로 명명하였다. R2A배지를 기본 배지로 하여 50 μg L-1 microcystin-LR을 첨가하여 30􀆆C, 12시간 동안 배양한 후 PPIA를 통해 microcystin이 80% 이상 분해되는 것을 확인하였다. Microcystin-LR의 분해를 HPLC 분석을 통해 재확인하였고, microcystin 분해산물로 추정되는 두 개의 peak를 확인하였다. 16S rRNA 염기서열을 이용한 계통분류 분석 결과, 본 연구에서 분리한 Microbacterium sp. MA21은 Alphaproteobacteria의 Sphingomonas 속에 속하지 않는 것은 물론 Actinobacteria에는 속하지만 기존에 보고되지 않은, 새로운 genus로 확인되었다.

1996년과 1997년에 남조류가 발생한 여러 호수에서 microcystin-RR, microcystin-LR, microcystin-YR의 함량을 정량하였고, 쥐를 이용하여 독성 검사를 실시하였다. 소양호를 제외한 모든 호수에서 Microcystis속의 종 들이 우점하였으며 전체 남조류 발생 호수의 시료에 대한 독소 검출의 빈도는 85%이였다. 조류세포내 총 microcystin의 함량은 0.29~2.61 mg/g이었으며 수중의 농도로 환산하면 0.4~21.6 μg/l이였다. 독소 함량은 1996년 보다 1997년에 높았고, 세 종류의 microcystin중 microcystin-RR이 가장 많이 검출되었다. 각 시료의 쥐에 대한 LD50은 총 microcystin함량과 LD50의 상관관계식에 의해 계산되었다. 화학분석에 의해 microcystin이 검출된 시료의 추출물이 주입되었을 때 쥐의 치사효과가 나타난 것으로 보아 microcystin 함량의 화학분석결 과와 bioassay 결과는 잘 일치하는 것으로 보이며, 영랑호 시료를 대상으로 실측한 LD50은 경험식으로 계산된 LD50과 일치하였다. 대상호수의 LD50은 27~338 mg/kg 의 범위로서 국내외에서 보고된 것에 비하여 높은 독성을 나타냈다.

The primary purpose of this study was to determine the risk of various disease outcomes due to exposure to cyanobacteria toxin (microcystin-LR) through drinking water in a Korean watershed. In order to determine the risk in a more quantitative way, the risk assessment framework developed by the National Research Council (NRC) of the United States (US) - hazard identification, dose-response relationship, exposure assessment, and risk characterization - was used in this study. For dose-response relationships, a computer software (BenchMark Dose Software (BMDS)) developed by the US Environmental Protection Agency (EPA) was used to fit the data from previous studies showing the relationship between the concentration of microcystin-LR and various disease outcomes into various dose-response models. For exposure assessment, the concentrations of microcystin-LR in the source water and finished water in a Korean watershed obtained from a recent study conducted by the Ministry of Environment of Korea were used. Finally, the risk of various disease outcomes due to exposure to cyanobacteria toxin (microcystin-LR) through drinking water was characterized by Monte-Carlo simulation using Crystall Ball program (Oracle Inc.) for adults and children. The results of this study suggest that the risk of disease due to microcystin-LR toxin through drinking water is very low and it appears that current water treatment practice should be able to protect the public from the harmful effects of cyanobacteria toxin (microcystin-LR) through drinking water.