Hassard Francis, Andrews Anthony, Jones Davey L, Parsons Louise, Jones Vera, Cox Brian A, Daldorph Peter, Brett Howard, McDonald James E, Malham Shelagh K
School of Ocean Sciences, Bangor University, Bangor, United Kingdom.
Cranfield Water Science Institute, Cranfield University, Bedford, United Kingdom.
Front Microbiol. 2017 Oct 17;8:1996. doi: 10.3389/fmicb.2017.01996. eCollection 2017.
To assess fecal pollution in coastal waters, current monitoring is reliant on culture-based enumeration of bacterial indicators, which does not account for the presence of viable but non-culturable or sediment-associated micro-organisms, preventing effective quantitative microbial risk assessment (QMRA). Seasonal variability in viable but non-culturable or sediment-associated bacteria challenge the use of fecal indicator organisms (FIOs) for water monitoring. We evaluated seasonal changes in FIOs and human enteric pathogen abundance in water and sediments from the Ribble and Conwy estuaries in the UK. Sediments possessed greater bacterial abundance than the overlying water column, however, key pathogenic species ( spp., spp., hepatitis A virus, hepatitis E virus and norovirus GI and GII) were not detected in sediments. was detected in low levels in the Conwy water in spring/summer and norovirus GII was detected in the Ribble water in winter. The abundance of and spp. quantified by culture-based methods, rarely matched the abundance of these species when measured by qPCR. The discrepancy between these methods was greatest in winter at both estuaries, due to low CFU's, coupled with higher gene copies (GC). Temperature accounted for 60% the variability in bacterial abundance in water in autumn, whilst in winter salinity explained 15% of the variance. Relationships between bacterial indicators/pathogens and physicochemical variables were inconsistent in sediments, no single indicator adequately described occurrence of all bacterial indicators/pathogens. However, important variables included grain size, porosity, clay content and concentrations of Zn, K, and Al. Sediments with greater organic matter content and lower porosity harbored a greater proportion of non-culturable bacteria (including dead cells and extracellular DNA) in winter. Here, we show the link between physicochemical variables and season which govern culturability of human enteric pathogens and FIOs. Therefore, knowledge of these factors is critical for accurate microbial risk assessment. Future water quality management strategies could be improved through monitoring sediment-associated bacteria and non-culturable bacteria. This could facilitate source apportionment of human enteric pathogens and FIOs and direct remedial action to improve water quality.
为评估沿海水域的粪便污染情况,当前的监测依赖于基于培养的细菌指标计数法,该方法无法检测到存活但不可培养或与沉积物相关的微生物,从而妨碍了有效的定量微生物风险评估(QMRA)。存活但不可培养或与沉积物相关的细菌的季节性变化,对将粪便指示生物(FIOs)用于水质监测提出了挑战。我们评估了英国里布尔河和康威河口的水和沉积物中FIOs以及人类肠道病原体丰度的季节性变化。沉积物中的细菌丰度高于上覆水柱,然而,在沉积物中未检测到关键致病物种( spp.、 spp.、甲型肝炎病毒、戊型肝炎病毒以及诺如病毒GI和GII)。春季/夏季在康威河水中检测到低水平的 ,冬季在里布尔河水中检测到诺如病毒GII。通过基于培养的方法定量的 和 spp. 的丰度,与通过定量聚合酶链反应(qPCR)测量时这些物种的丰度很少匹配。在两个河口,冬季这两种方法之间的差异最大,原因是菌落形成单位(CFU)较低,同时基因拷贝数(GC)较高。秋季,温度占水中细菌丰度变异性的60%,而冬季盐度解释了15%的方差。在沉积物中,细菌指标/病原体与理化变量之间的关系不一致,没有单一指标能充分描述所有细菌指标/病原体的出现情况。然而,重要的变量包括粒度、孔隙率、粘土含量以及锌、钾和铝的浓度。在冬季,有机质含量较高且孔隙率较低的沉积物中,不可培养细菌(包括死细胞和细胞外DNA)的比例更大。在此,我们展示了理化变量与季节之间的联系,它们决定了人类肠道病原体和FIOs的可培养性。因此,了解这些因素对于准确的微生物风险评估至关重要。通过监测与沉积物相关的细菌和不可培养细菌,未来的水质管理策略可能会得到改进。这有助于对人类肠道病原体和FIOs进行源分配,并指导采取补救行动以改善水质。