Yuan Jian, Kim Hyun-Joong, Filstrup Christopher T, Guo Baoqing, Imerman Paula, Ensley Steve, Yoon Kyoung-Jin
Departments of Veterinary Diagnostic and Production Animal Medicine (Yuan, Guo, Imerman, Ensley, Yoon), Iowa State University, Ames, IA.
Food Science and Human Nutrition (Kim), Iowa State University, Ames, IA.
J Vet Diagn Invest. 2020 May;32(3):369-381. doi: 10.1177/1040638720916156. Epub 2020 Apr 20.
is a widespread freshwater cyanobacterium that can produce microcystin, a potent hepatotoxin harmful to animals and humans. Therefore, it is crucial to monitor for the presence of toxigenic spp. to provide early warning of potential microcystin contamination. Microscopy, which has been used traditionally to identify spp., cannot differentiate toxigenic from non-toxigenic . We developed a PCR-based method to detect toxigenic spp. based on detection of the microcystin synthetase C () gene and 16S rRNA gene. Specificity was validated against toxic and nontoxic strains, as well as 4 intergeneric freshwater cyanobacterial strains. Analytical sensitivity was as low as 747 fg/µL genomic DNA (or 3 cells/µL) for toxic . Furthermore, we tested 60 water samples from 4 farm ponds providing drinking water to swine facilities in the midwestern United States using this method. Although all water samples were positive for spp. (i.e., 16S rRNA gene), toxigenic spp. were detected in only 34 samples (57%). Seventeen water samples contained microcystin (0.1-9.1 μg/L) determined with liquid chromatography-mass spectrometry, of which 14 samples (82%) were positive for . A significant correlation was found between the presence of toxigenic spp. and microcystin in water samples ( = 0.0004). Our PCR method can be a low-cost molecular tool for rapid and specific identification of toxigenic spp. in farm ponds, improving detection of microcystin contamination, and ensuring water safety for farm animals.
是一种广泛分布的淡水蓝藻细菌，可产生微囊藻毒素，这是一种对动物和人类有害的强效肝毒素。因此，监测产毒 属物种的存在对于提供潜在微囊藻毒素污染的早期预警至关重要。传统上用于鉴定 属物种的显微镜检查无法区分产毒和非产毒 。我们开发了一种基于聚合酶链反应（PCR）的方法，基于对微囊藻毒素合成酶C（ ）基因和16S核糖体RNA（rRNA）基因的检测来检测产毒 属物种。针对有毒和无毒 菌株以及4种淡水蓝藻细菌属间菌株验证了特异性。对于有毒 ，分析灵敏度低至747 fg/µL基因组DNA（或3个细胞/µL）。此外，我们使用此方法测试了来自美国中西部为养猪场提供饮用水的4个农场池塘的60份水样。尽管所有水样的 属物种（即16S rRNA基因）均呈阳性，但仅在34份样品（57%）中检测到产毒 属物种。用液相色谱-质谱法测定的17份水样含有微囊藻毒素（0.1 - 9.1 μg/L），其中14份样品（82%）的 呈阳性。在水样中产毒 属物种的存在与微囊藻毒素之间发现了显著相关性（ = 0.0004）。我们的PCR方法可以成为一种低成本的分子工具，用于快速、特异性地鉴定农场池塘中的产毒 属物种，改进微囊藻毒素污染的检测，并确保农场动物的用水安全。
