Saker M L, Jungblut A-D, Neilan B A, Rawn D F K, Vasconcelos V M
Department of Ecotoxicology, Centro Interdisciplinar de Investigação Marinha e Ambiental, Rua dos Bragas 289, 4050 Porto, Portugal.
Toxicon. 2005 Oct;46(5):555-62. doi: 10.1016/j.toxicon.2005.06.021.
In this study we investigated the presence of toxin-producing cyanobacterial contaminants in food supplements manufactured from blooms of the non-toxic freshwater cyanobacterium Aphanizomenon flos-aquae. Previous reports investigating the contamination of health food supplements with toxin-producing cyanobacteria have used chemical and or biochemical methods such as HPLC, ELISA and protein phosphatase assays. Whilst these studies have drawn attention to the presence of hepatotoxic microcystins in some commercially available food supplements, the methods used do not provide any information on the source of the contaminant. Such information would be useful for the quality control of food supplements produced for human consumption. In this study we applied a molecular technique, involving the amplification of the 16s rRNA gene, the phycocyanin operon, and two genes of the microcystin synthetase gene cluster to show that all 12 food supplement samples, sourced from various internet distributors and containing non-toxic A. flos-aquae, also contained toxigenic cyanobacteria. Sequencing of the microcystin synthetase genes detected in all of the food supplements showed that M. aeruginosa was the organism responsible for the production of microcystins in the samples. The presence of microcystins in the food supplements was confirmed by ELISA, with concentrations within the range of 0.1--4.72 microgg(-1) (microcystin-LR equivalents). Given that the molecular methods applied here are highly sensitive, and show good agreement with the results obtained from ELISA, we believe that they could potentially be used as a quality control technique for food products that contain cyanobacteria.
在本研究中,我们调查了由无毒淡水蓝藻水华鱼腥藻制成的食品补充剂中产生毒素的蓝藻污染物的存在情况。先前有关产毒蓝藻对健康食品补充剂污染的报告使用了化学和/或生化方法,如高效液相色谱法(HPLC)、酶联免疫吸附测定法(ELISA)和蛋白磷酸酶测定法。虽然这些研究已关注到一些市售食品补充剂中存在肝毒性微囊藻毒素,但所用方法并未提供有关污染物来源的任何信息。此类信息对于生产供人类食用的食品补充剂的质量控制将是有用的。在本研究中,我们应用了一种分子技术,涉及16s rRNA基因、藻蓝蛋白操纵子以及微囊藻毒素合成酶基因簇的两个基因的扩增,以表明所有12个食品补充剂样品,这些样品购自不同的网络经销商且含有无毒的水华鱼腥藻,也含有产毒蓝藻。对在所有食品补充剂中检测到的微囊藻毒素合成酶基因进行测序表明,铜绿微囊藻是样品中产生微囊藻毒素的生物体。通过ELISA确认了食品补充剂中微囊藻毒素的存在,其浓度范围为0.1 - 4.72微克/克(微囊藻毒素-LR当量)。鉴于此处应用的分子方法高度灵敏,且与ELISA获得的结果显示出良好的一致性,我们认为它们有可能用作含有蓝藻的食品的质量控制技术。