Aniline and parachloroaniline (PCA) are used in manufacturing industries. Aniline and PCA are toxic and suspected carcinogenic pollutants. These compounds have entered the environment and are problematic groundwater pollutants. We are evaluating the potential for aniline and p-chloroaniline biodegradation by native aquifer microorganisms from a large, chemically and geologically complex industrial site historically contaminated with many different chemicals including aniline and chlorinated aniline. A 55-ft sediment core was recovered from the site along with groundwater from a multi-level sampler. The sediment core was divided into 2-inch slices. The ~300 sediment core slices were shared among multiple teams and characterized for in situ geochemistry and microbial community via next generation sequencing of 16S rRNA genes. We used this data to develop hypotheses for microbial activity and simulating in situ conditions. Sediments from a contaminated site were enriched with aniline and PCA as carbon sources. Two groups of microcosms, targeted and composited, were established. Aerobic, sulfate-, nitrate-, and iron-reducing microcosms were established and incubated in the dark at room temperature. Non-active microcosms were bioaugmented using pure isolate obtained from the site. Microcosms were ~15% sediment slurry in groundwater from a matching depth or in minimal medium. Autoclaved controls were also established. Experiments were performed under sterile conditions using triplicates. Microcosms were amended with aniline and PCA and monitored by HPLC. Active microcosms were plated (0.1 mL) on minimal media agar plates without carbon and nitrogen sources to isolate pure cultures. Community analysis was done for the active aerobic microcosms. The targeted microcosms showed no substantial loss of aniline or p-aniline. The composited microcosms from the -9.2 to 10 ft depth showed a complete loss of aniline and some loss of p-aniline compared to the killed controls. Rhodococcus HA1 was isolated from the aerobic microcosms. HA1 uses aniline as a carbon and nitrogen source. Aniline was depleted in two weeks in non-active microcosms after bioaugmentation with pure isolates. Analysis of 16S rRNA genes showed a shift in the microbial community at the genus level in active microcosms in comparison to the original community. On-going work is focused on use of stable isotope probing (SIP) to determine which organisms are active in the microcosm bacterial community.

Biodegradation of aniline and p-chloroaniline under different redox conditions at a contaminated chemical manufacturing site

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.