Ranchou-Peyruse Magali, Guignard Marion, Haddad Perla G, Robin Sylvain, Boesch Fabrice, Lanot Maud, Carrier Hervé, Dequidt David, Chiquet Pierre, Caumette Guilhem, Cézac Pierre, Ranchou-Peyruse Anthony
E2S-UPPA, CNRS, IPREM, Universite de Pau & Pays Adour, Pau, France.
E2S-UPPA, LaTEP, Universite de Pau & Pays Adour, Pau, France.
Front Microbiol. 2023 Jan 4;13:1012400. doi: 10.3389/fmicb.2022.1012400. eCollection 2022.
To be effective, microbiological studies of deep aquifers must be free from surface microbial contaminants and from infrastructures allowing access to formation water (wellheads, well completions). Many microbiological studies are based on water samples obtained after rinsing a well without guaranteeing the absence of contaminants from the biofilm development in the pipes. The protocol described in this paper presents the adaptation, preparation, sterilization and deployment of a commercial downhole sampler (PDSshort, Leutert, Germany) for the microbiological studying of deep aquifers. The ATEX sampler (i.e., explosive atmospheres) can be deployed for geological gas storage (methane, hydrogen). To validate our procedure and confirm the need to use such a device, cell counting and bacterial taxonomic diversity based on high-throughput sequencing for different water samples taken at the wellhead or at depth using the downhole sampler were compared and discussed. The results show that even after extensive rinsing (7 bore volumes), the water collected at the wellhead was not free of microbial contaminants, as shown by beta-diversity analysis. The downhole sampler procedure was the only way to ensure the purity of the formation water samples from the microbiological point of view. In addition, the downhole sampler allowed the formation water and the autochthonous microbial community to be maintained at pressure for laboratory analysis. The prevention of the contamination of the sample and the preservation of its representativeness are key to guaranteeing the best interpretations and understanding of the functioning of the deep biosphere.
为确保有效性,深层含水层的微生物学研究必须避免受到地表微生物污染物以及能接触到地层水的基础设施(井口、完井装置)的影响。许多微生物学研究基于对水井进行冲洗后采集的水样,但无法保证管道中生物膜生长产生的污染物不存在。本文所述方案介绍了一种商用井下采样器(PDSshort,德国勒特)用于深层含水层微生物学研究的改装、制备、灭菌及部署。该ATEX采样器(即适用于爆炸环境)可用于地质储气库(甲烷、氢气)。为验证我们的程序并确认使用此类装置的必要性,我们对使用井下采样器在井口或深部采集的不同水样,基于高通量测序进行细胞计数和细菌分类多样性分析,并进行了比较和讨论。结果表明,即使经过大量冲洗(7倍井筒容积),通过β多样性分析仍表明井口采集的水并非没有微生物污染物。从微生物学角度来看,井下采样器程序是确保地层水样品纯净的唯一方法。此外,井下采样器能使地层水和本地微生物群落保持压力以便进行实验室分析。防止样品污染并保持其代表性是保证对深层生物圈功能进行最佳解读和理解的关键。