Effects of produced water from oil drilling on pregnant and not pregnant mice and their pups microbiome across multiple gestation periods using Nanopore Technologies Sequencing
Produced water is a wastewater byproduct of oil and gas extraction, containing a complex mixture of formation fluid and chemical additives used in production and maintenance. In our area of concern, the National Pollutant Discharge Elimination System permits the release of this water for livestock and wildlife watering, including the irrigation of crops. Little is known about the impacts of this water on humans and surrounding wildlife, therefore this study (as well as others in a larger project) was run to determine the impacts of produced water on mice gut microbiomes. In this project, pregnant and not pregnant mice (strain C578L/6N) were exposed to produced water at varying concentrations (0% (lab water), 12.5%, 50%, and 100%) through the drinking water (sipper sacks). Over the course of the study no changes in body weight gains, organ weights, or histopath changes occurred. Caecum samples were removed from the mice post mortum, both the dams and their pups (the pups were never introduced to the produced water except through their mothers). Microbiome DNA was isolated from the whole caecum samples (only the 0% and 100% for the dams and 0%, 50%, and 100% for the pups) using the Qiagen DNeasy PowerSoil Pro Kit according to the manufacturer's instructions. After isolation DNA concentration and quality was quantified using a Qubit Flex Fluorometer, Nanodrop 2000c Spectrophotometer, and an Agilent Bioanalyzer System. 16S rDNA was amplified using the Oxford Nanopore Technologies 16S Barcoding Kit. The DNA was amplified by PCR using specific 16S primers, 27F adn 1492R. The amplified DNA was sequenced using a R9.4.1 flow cell on the Oxford Nanopore Technologies MinION sequencer.