Climate Change Cluster Faculty of Science, University of Technology Sydney, Sydney, NSW, Australia.
Waters, Wetlands, Coasts Science Branch, NSW Department of Primary Industries and Environment, Lidcombe, NSW, 2141, Australia.
Water Res. 2022 Jun 30;218:118534. doi: 10.1016/j.watres.2022.118534. Epub 2022 Apr 30.
Urbanised beaches are regularly impacted by faecal pollution, but management actions to resolve the causes of contamination are often obfuscated by the inability of standard Faecal Indicator Bacteria (FIB) analyses to discriminate sources of faecal material or detect other microbial hazards, including antibiotic resistance genes (ARGs). We aimed to determine the causes, spatial extent, and point sources of faecal contamination within Rose Bay, a highly urbanised beach within Sydney, Australia's largest city, using molecular microbiological approaches. Sampling was performed across a network of transects originating at 9 stormwater drains located on Rose Bay beach over the course of a significant (67.5 mm) rainfall event, whereby samples were taken 6 days prior to any rain, on the day of initial rainfall (3.8 mm), three days later after 43 mm of rain and then four days after any rain. Quantitative PCR (qPCR) was used to target marker genes from bacteria (i.e., Lachnospiraceae and Bacteroides) that have been demonstrated to be specific to human faeces (sewage), along with gene sequences from Heliobacter and Bacteriodes that are specific to bird and dog faeces respectively, and ARGs (sulI, tetA, qnrS, dfrA1 and vanB). 16S rRNA gene amplicon sequencing was also used to discriminate microbial signatures of faecal contamination. Prior to the rain event, low FIB levels (mean: 2.4 CFU/100 ml) were accompanied by generally low levels of the human and animal faecal markers, with the exception of one transect, potentially indicative of a dry weather sewage leak. Following 43 mm of rain, levels of both human faecal markers increased significantly in stormwater drain and seawater samples, with highest levels of these markers pinpointing several stormwater drains as sources of sewage contamination. During this time, sewage contamination was observed up to 1000 m from shore and was significantly and positively correlated with often highly elevated levels of the ARGs dfrA1, qnrS, sulI and vanB. Significantly elevated levels of the dog faecal marker in stormwater drains at this time also indicated that rainfall led to increased input of dog faecal material from the surrounding catchment. Using 16S rRNA gene amplicon sequencing, several indicator taxa for stormwater contamination such as Arcobacter spp. and Comamonadaceae spp. were identified and the Bayesian SourceTracker tool was used to model the relative impact of specific stormwater drains on the surrounding environment, revealing a heterogeneous contribution of discrete stormwater drains during different periods of the rainfall event, with the microbial signature of one particular drain contributing up to 50% of bacterial community in the seawater directly adjacent. By applying a suite of molecular microbiological approaches, we have precisely pinpointed the causes and point-sources of faecal contamination and other associated microbiological hazards (e.g., ARGs) at an urbanised beach, which has helped to identify the most suitable locations for targeted management of water quality at the beach.
城市海滩经常受到粪便污染的影响,但由于标准粪便指示细菌(FIB)分析无法区分粪便来源或检测其他微生物危害,包括抗生素耐药基因(ARGs),因此解决污染原因的管理措施常常变得复杂。我们的目的是使用分子微生物学方法确定澳大利亚最大城市悉尼的高度城市化海滩玫瑰湾内粪便污染的原因、空间范围和点源,使用分子微生物学方法。在一次重大(67.5 毫米)降雨事件过程中,在位于玫瑰湾海滩上的 9 个雨水渠的网络上进行了采样,在降雨前 6 天、初始降雨(3.8 毫米)当天、降雨后 3 天进行了采样,43 毫米降雨后 3 天,以及降雨后 4 天。定量 PCR(qPCR)用于针对细菌(即lachnospiraceae 和 bacteroides)的标记基因进行靶向,这些基因已被证明是人类粪便(污水)特异性的,以及针对鸟类和犬类粪便特异性的 helicobacter 和 bacteriodes 的基因序列,以及抗生素耐药基因(sulI、tetA、qnrS、dfrA1 和 vanB)。16S rRNA 基因扩增子测序也用于区分粪便污染的微生物特征。在降雨事件之前,低 FIB 水平(平均值:2.4 CFU/100 ml)伴随着人类和动物粪便标记物的一般低水平,除了一个横断面,可能表明有干燥天气污水泄漏。在 43 毫米降雨后,雨水渠和海水中的两种人类粪便标记物水平显著增加,这些标记物的最高水平指出了几个雨水渠是污水污染的来源。在此期间,污水污染一直延伸到离海岸 1000 米的地方,并且与抗生素耐药基因 dfrA1、qnrS、sulI 和 vanB 的水平高度升高呈显著正相关。此时,雨水渠中犬类粪便标记物的显著升高也表明,降雨导致周围集水区犬类粪便物质的输入增加。使用 16S rRNA 基因扩增子测序,鉴定了一些雨水渠污染的指示性分类群,如 Arcobacter spp.和 Comamonadaceae spp.,并使用贝叶斯源追踪工具对特定雨水渠对周围环境的相对影响进行建模,揭示了不同降雨事件期间离散雨水渠的不均匀贡献,一个特定雨水渠的微生物特征直接贡献了相邻海水中细菌群落的 50%。通过应用一系列分子微生物学方法,我们精确地确定了城市化海滩的粪便污染和其他相关微生物危害(例如抗生素耐药基因)的原因和点源,并有助于确定海滩水质有针对性管理的最合适位置。