Microbial Community and Atrazine-Degrading Genetic Potential in Deep Zones of a Hypersaline Lake-Aquifer System

Abstract

The chemical composition of groundwater and related surface water ecosystems can be modified by intensive agricultural activities. This is the case of the Natural Reserve of Pétrola saline lake (Albacete, SE Spain), which constitutes the discharge area of an unconfined aquifer. The extended use of fertilizers and pesticides poses a threat to ecosystem sustainability. One of the most applied herbicides worldwide has been atrazine. Despite being prohibited in Spain since 2007 by European regulations, atrazine can still be detected due to its high persistence in the environment. Atrazine degradation pathways are mediated by biological processes performed by microorganisms with adapted metabolic mechanisms that make in situ bioremediation possible. To evaluate the presence of such microorganisms in the unconfined aquifer, groundwater was collected from a flowing 37.9 m deep piezometer. DNA was extracted, and the bacterial 16S rRNA gene was amplified and cloned. Later, 93 clones were sequenced, providing the first molecular assessment of bacterial community structure in the deep zones of the aquifer. Some of these bacteria have been previously described to be involved in atrazine degradation. In addition, 14 bacteria were isolated from the groundwater samples and identified by 16S rRNA gene sequencing. DNA from these bacteria was subjected to PCR assays with primers designed for the genes involved in the atrazine degradation pathway. Positive results in the amplification were found in at least three of these bacteria (Arthrobacter sp., Nocardioides sp. and Pseudomonas sp.). The atrazine-degrading genetic potential was shown to be dependent on the trzN and atzA,B,C gene combination. These results suggest for the first time the adaptation of the bacterial population present in deep aquifer zones to atrazine exposure, even after more than 15 years of its ban in Spain. In addition, this study provides the baseline data about the bacterial communities found in deep aquifer zones from the hypersaline lake-aquifer system.