PDF(Pubmed)
Abstract：
Harmful algal bloom (HAB) formation leads to the eutrophication of water ecosystems and may render recreational lakes unsuitable for human use. We evaluated the applicability and comparison of metabarcoding, metagenomics, qPCR, and ELISA-based methods for cyanobacteria/cyanotoxin detection in bloom and non-bloom sites for the Great Lakes region. DNA sequencing-based methods robustly identified differences between bloom and non-bloom samples (e.g., the relative prominence of Anabaena and Planktothrix). Shotgun sequencing strategies also identified the enrichment of metabolic genes typical of cyanobacteria in bloom samples, though toxin genes were not detected, suggesting deeper sequencing or PCR methods may be needed to detect low-abundance toxin genes. PCR and ELISA indicated microcystin levels and microcystin gene copies were significantly more abundant in bloom sites. However, not all bloom samples were positive for microcystin, possibly due to bloom development by non-toxin-producing species. Additionally, microcystin levels were significantly correlated (positively) with microcystin gene copy number but not with total cyanobacterial 16S gene copies. In summary, next-generation sequencing-based methods can identify specific taxonomic and functional targets, which can be used for absolute quantification methods (qPCR and ELISA) to augment conventional water monitoring strategies.
摘要：
有害藻华（HAB）的形成导致水生态系统的富营养化，并可能使休闲湖泊不适合人类使用。我们评估了元编码的适用性和比较，宏基因组学，qPCR,以及基于ELISA的方法，用于检测大湖地区的水华和非水华地区的蓝细菌/蓝毒素。基于DNA测序的方法强有力地识别了布卢姆和非布卢姆样品之间的差异(例如，鱼腥草和浮游植物的相对突出)。Shotgun测序策略还确定了水华样品中典型的蓝藻代谢基因的富集，尽管没有检测到毒素基因，提示可能需要更深入的测序或PCR方法来检测低丰度的毒素基因。PCR和ELISA表明，盛开部位的微囊藻毒素水平和微囊藻毒素基因拷贝明显更丰富。然而,并非所有的水华样本都对微囊藻毒素呈阳性,可能是由于不产生毒素的物种的开花发展。此外,微囊藻毒素水平与微囊藻毒素基因拷贝数显着相关（正相关），但与蓝藻16S基因总拷贝数无关。总之,基于下一代测序的方法可以识别特定的分类和功能靶标，可用于绝对定量方法（qPCR和ELISA）以增强常规的水监测策略。
