Cawthron Institute, Private Bag 2, Nelson 7010, New Zealand.
Cawthron Institute, Private Bag 2, Nelson 7010, New Zealand.
Harmful Algae. 2020 Jul;97:101869. doi: 10.1016/j.hal.2020.101869. Epub 2020 Jul 9.
Toxic cyanobacterial blooms are becoming more prevalent in freshwater systems, increasing the need for monitoring to protect human health. Phycocyanin fluorescence sensors have been developed as tools for providing fast and cost-effective proxy measurements for cyanobacterial biomass. However, poor precision and low sensitivity in many of the probe sensors assessed to-date has restricted their potential for practical application in cyanobacterial monitoring programmes. In the present study, the sensitivity and accuracy of a handheld fluorometer, the CyanoFluor, was assessed using 12 cyanobacterial strains and samples from four different lakes collected weekly for 12 weeks. After the initial measurements, the samples were lysed by sonication, which we hypothesised would reduce inter and intra-specific differences. The CyanoFluor displayed high sensitivity (limit of quantification = 3.5 µg L of phycocyanin) and was able to detect cyanobacterial biovolumes to levels much lower than the threshold levels in current recreational guidelines worldwide. There were strong and significant phycocyanin to biovolume relationships (r ≥ 0.88, P < 0.05) for all 12 cyanobacterial cultures. Collectively, strong relationships between phycocyanin fluorescence and cyanobacterial biovolumes were also identified in environmental samples (r ≥ 0.78, P < 0.001), although weaker relationships were identified when lakes were analysed separately (r = 0.06 - 0.90). There were differences in phycocyanin per biovolume between both cultured strains and lakes, highlighting innate interspecific differences that exist between cyanobacterial species. Lysis of samples consistently reduced variability between technical replicates, in cyanobacteria cultures (up to 87% reduction in sample variability) and environmental samples (71 - 93% reduction), indicating that it would be a useful methodological step to improve the repeatability of results. When guideline thresholds (aligned with currently enforced risk assessment categories) were modelled based on the most successful linear regression model, 74% of samples were assigned to the correct risk category. The sensitivity of the CyanoFluor and accuracy of the phycocyanin threshold models, indicates high potential for this method to be integrated into cyanobacterial monitoring programmes.
有毒蓝藻水华在淡水系统中越来越普遍,因此需要进行监测以保护人类健康。藻青蛋白荧光传感器已被开发为提供快速且具有成本效益的蓝藻生物量替代测量方法的工具。然而,迄今为止评估的许多探头传感器的精度和灵敏度都很低,限制了它们在蓝藻监测计划中的实际应用潜力。在本研究中,使用 12 种蓝藻菌株和来自四个不同湖泊的每周采集一次的样本,评估了手持荧光计 CyanoFluor 的灵敏度和准确性。初始测量后,通过超声处理裂解样品,我们假设这将减少种内和种间差异。CyanoFluor 具有高灵敏度(定量下限= 3.5μg L 藻青蛋白),能够检测到比当前全球休闲指南中的阈值水平低得多的蓝藻生物量。所有 12 种蓝藻培养物的藻青蛋白与生物量之间都存在强而显著的关系(r≥0.88,P<0.05)。在环境样本中也确定了藻青蛋白荧光与蓝藻生物量之间的强关系(r≥0.78,P<0.001),尽管当分别分析湖泊时,关系较弱(r=0.06-0.90)。在培养菌株和湖泊之间,单位生物量的藻青蛋白含量存在差异,这突出了蓝藻物种之间存在的固有种间差异。样品的裂解一致降低了蓝藻培养物(样品变异性降低高达 87%)和环境样品(71-93%的降低)之间技术重复的可变性,表明这将是提高结果可重复性的有用方法步骤。当基于最成功的线性回归模型模拟指南阈值(与当前执行的风险评估类别对齐)时,74%的样本被分配到正确的风险类别。CyanoFluor 的灵敏度和藻青蛋白阈值模型的准确性表明,该方法具有很高的潜力可集成到蓝藻监测计划中。