Gkelis S, Lanaras T, Sivonen K
Department of Botany, P.O. Box 109, GR-541 24, Aristotle University of Thessaloniki, Greece.
Aquat Toxicol. 2006 Jun 10;78(1):32-41. doi: 10.1016/j.aquatox.2006.02.001. Epub 2006 Mar 15.
Toxic bloom-forming cyanobacteria can cause animal death and adversely affect human health. Blooms may contain microcystins (MCs), cyanobacterial heptapeptide hepatotoxins and other peptides such as anabaenopeptins and anabaenopeptilides. MCs have been shown to occur in various aquatic organisms including mussels, water snails, crustaceans and fish. Muscle and viscera samples from eight species of fish (Acipenser gueldenstaedtii, Carassius auratus, Carassius gibelio, Cyprinus carpio, Perca fluviatilis, Rutilus rubilio, Silurus aristotelis and Silurus glanis), a frog (Rana eperotica), a mussel (Anodonta sp.) and a water snail (Viviparus contectus) were analyzed by high-performance liquid chromatography (HPLC), protein phosphatase 1 (PP1) inhibition assay (PP1IA) and ELISA. MC(s) was detected in all fish, frog, mussel and water snail samples tested by PP1IA and ELISA, including the frog R. eperotica and the freshwater snail V. contectus, in which the occurrence of MCs was not previously known. MC concentration ranged from 20 to 1500 ng g(-1)dw and from 25 to 5400 ng g(-1)dw in muscle and visceral tissue of fishes and frogs, respectively. In mussel and water snail tissue MC concentration ranged from 1650 to 3495 ng g(-1)dw. HPLC analysis revealed peaks having the same UV spectrum as anabaenopeptin- or anabaenopeptilide-like compounds, not previously known to occur in aquatic fauna tissue. The concentrations of the compounds detected ranged from 1.5 to 230 microg g(-1)dw. Comparison of the PP1IA and ELISA showed that values obtained with PP1IA where higher than those obtained with ELISA. Anabaenopeptins and/or anabaenopeptilides occurring in faunal tissue may account for the higher PP1IA values as we found that PP1 activity was inhibited by the purified anabaenopeptins A (45-60% inhibition) and B (5-75% inhibition). Purified anabaenopeptilides 90A and 90B exhibited weaker PP1 inhibition activity (5-35 and 5-23% inhibition, respectively). This is the first report of MC occurrence in aquatic animals collected from freshwaters of southern Europe.
形成有毒水华的蓝藻可导致动物死亡，并对人类健康产生不利影响。水华可能含有微囊藻毒素（MCs）、蓝藻七肽肝毒素以及其他肽类，如鱼腥藻肽和鱼腥藻肽内酯。已证明MCs存在于各种水生生物中，包括贻贝、田螺、甲壳类动物和鱼类。采用高效液相色谱法（HPLC）、蛋白磷酸酶1（PP1）抑制试验（PP1IA）和酶联免疫吸附测定法（ELISA）对8种鱼类（史氏鲟、鲫鱼、银鲫、鲤鱼、河鲈、赤梢鱼、斑真鮰和欧洲六须鲶）、1只青蛙（沼蛙）、1只贻贝（无齿蚌属）和1只田螺（静水椎实螺）的肌肉和内脏样本进行了分析。通过PP1IA和ELISA检测，在所有测试的鱼类、青蛙、贻贝和田螺样本中均检测到了MCs，包括此前未知有MCs存在的沼蛙和淡水螺静水椎实螺。鱼类和青蛙肌肉及内脏组织中的MC浓度分别为20至1500 ng g(-1)干重和25至5400 ng g(-1)干重。贻贝和田螺组织中的MC浓度为1650至3495 ng g(-1)干重。HPLC分析显示出与鱼腥藻肽或鱼腥藻肽内酯类化合物具有相同紫外光谱的峰，此前在水生动物组织中尚未发现此类化合物。检测到的这些化合物浓度为1.5至230 μg g(-1)干重。PP1IA和ELISA的比较表明，PP1IA获得的值高于ELISA获得的值。动物组织中出现的鱼腥藻肽和/或鱼腥藻肽内酯可能是PP1IA值较高的原因，因为我们发现纯化的鱼腥藻肽A（抑制率45 - 60%）和B（抑制率5 - 75%）可抑制PP1活性。纯化的鱼腥藻肽内酯90A和90B表现出较弱的PP1抑制活性（分别为5 - 35%和5 - 23%抑制率）。这是关于在欧洲南部淡水采集的水生动物中检测到MCs的首次报告。
