An epidemiological survey for the causes of a high incidence of primary liver cancer (PLC) in Haimen city, Jian-Su province and Fusui county, Guangxi province in China, found a close correlation between the incidence of PLC and the drinking of pond and ditch water. With an aim to clarify whether microcystins (MC), a hepatotoxic peptide produced by water bloom algae, contaminate the drinking water in the endemic areas of PLC in China, a highly sensitive enzyme-linked immunosorbent assay with a detection limit of 50 pg/ml, was introduced to monitor the MC. Three trials to survey the drinking water were carried out in 1993-1994. Samples, 1135 in total, were collected from different sources such as: ponds, ditches, rivers, shallow wells and deep wells in Haimen city. The first survey in September 1993 found that three out of 14 ditch water specimens were positive for MC, with a range of 90-460 pg/ml. Several toxic algae such as Oscillatoria agardhii were present in some of the ditches. In the second trial, samples were collected from five ponds/ditches, two rivers, two shallow wells and two deep wells monthly for the whole year of 1994. These data showed that MC was highest in June to September, with a range of 62-296 pg/ml. A third trial on the 989 different water samples collected from the different types of water sources in July 1994 revealed that 17% of the pond/ditch water, 32% of the river water, and 4% of the shallow-well water were positive for MC, with averages of 101, 160 and 68 pg/ml respectively. No MC was detected in deep well water. A similar survey on 26 drinking water samples in Fusui, Guangxi province, demonstrated a high contamination frequency of MC in the water of ponds/ditches and rivers but no MC in shallow and deep wells. These data support a hypothesis that the blue-green algal toxin MC in the drinking water of ponds/ditches and rivers, or both, is one of the risk factors for the high incidence of PLC in China. Based on previous findings on the epidemiology of PLC and the present results from the mass screening of MC in the drinking water, an advisory level of MC in drinking water was proposed to below 0.01 microg/l. The combined effect of a potent hepatocarcinogen AFB1 and an intermittent intake of MC in drinking water in the summer season was discussed as an etiology of PLC.

Microcystins and nodularin, isolated from toxic blue-green algae, are hepatotoxic monocyclic polypeptides. Both microcystins and nodularin inhibited in vitro protein phosphatase activity present in a cytosolic fraction of mouse liver, bound to the okadaic acid receptors, protein phosphatases 1 and 2A, and thus resulted in the increase of phosphoproteins; this was referred to as the apparent "activation" of protein kinases. Their concentrations causing 50% of the maximal effects are comparable to that of okadaic acid, a potent protein phosphatase inhibitor and a potent tumor promoter, in the nanomolar range of concentration. The increase of phosphoproteins was observed in rat primary cultured hepatocytes and was subsequently associated with morphological changes, which appeared to be a step in the process of hepatotoxicity. The well-known hepatotoxic compounds, alpha-amanitin and phalloidin, did not show any effects similar to those of microcystins, nodularin and okadaic acid. It is suggested that the hepatotoxicity of microcystins and nodularin may result from inhibition of protein phosphatases and the increase of phosphoproteins.

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Effects of light intensity, temperature, and nutrients on the toxicity of Microcystis aeruginosa were investigated, using a toxic strain which kills mice. A marked change in toxicity was observed in the light intensity experiment, and slight changes were observed to be caused by temperature and phosphorus deficiency.

Temporal variability in the concentration of toxic heptapeptide microcystin was studied ( ) ( ) during the warm season of four years 1991 -1994 in a hypertrophic lake Lake Suwa in central ( ) ( Honshu, Japan. Lake water samples ca. 5 L were filtered to separate intracellular microcystin cell ) ( ) fraction from extracellular microcystin filtered lake water fraction . These fractions were analyzed to measure the total quantity of microcystin in lake water. Total amounts of extra-and intracellular microcystin were measured with high performance liquid chromatography. Concentrations of intracellu-( ) lar microcystin usually exceeded concentrations of extracellular microcystin 24 out of 26 times . High concentrations of intracellular microcystin were found during the exponential growth phase of the blooms, whereas concentrations of extracellular microcystin were highest at the end of the blooms.( ) However, concentrations of extracellular microcystin remained very small -4 g / L compared to the levels of intracellular microcystin. The relatively higher percentages of microcystin in filtered lake water ( ) )20% at the end of blooms suggests that release of microcystin from cells occurs during senescence and the decomposition period of Microcystis cells.