Abstract

BackgroundBioaugmentation aims to use the capacities of specific bacterial strains inoculated into sites to enhance pollutant biodegradation. Bioaugmentation results have been mixed, which has been attributed to poor inoculant growth and survival in the field, and, consequently, moderate catalytic performance. However, our understanding of biodegradation activity mostly comes from experiments conducted under laboratory conditions, and the processes occurring during adaptation and invasion of inoculants into complex environmental microbiomes remain poorly known. The main aim of this work was thus to study the specific and different cellular reactions of an inoculant for bioaugmentation during adaptation, growth and survival in natural clean and contaminated non-sterile soils, in order to better understand factors limiting bioaugmentation.ResultsAs inoculant we focused on the monoaromatic compound-degrading bacterium Pseudomonas veronii 1YdBTEX2. The strain proliferated in all but one soil types in presence and in absence of exogenously added toluene. RNAseq and differential genome-wide gene expression analysis illustrated both a range of common soil responses such as increased nutrient scavenging and recycling, expression of defense mechanisms, as well as environment-specific reactions, notably osmoprotection and metal homeostasis. The core metabolism of P. veronii remained remarkably constant during exponential growth irrespective of the environment, with slight changes in cofactor regeneration pathways, possibly needed for balancing defense reactions.ConclusionsP. veronii displayed a versatile global program, enabling it to adapt to a variety of soil environments in the presence and even in absence of its target pollutant toluene. Our results thus challenge the widely perceived dogma of poor survival and growth of exogenous inoculants in complex microbial ecosystems such as soil and provide a further basis to developing successful bioaugmentation strategies.

Highlights

  • Bioaugmentation aims to use the capacities of specific bacterial strains inoculated into sites to enhance pollutant biodegradation

  • To more count P. veronii colony-forming units (CFUs) in soil, we used a derivative with a miniTn5 insertion constitutively expressing the green fluorescent protein (GFP) from the Pcirc promoter of the ICEclc element [24]

  • Comparative growth of P. veronii in different soils with toluene as added carbon substrate In order to benchmark growth of P. veronii in different soils, we inoculated microcosms dosed with toluene in comparison to liquid suspended medium or to artificial porous medium (APM, Fig. 1)

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Summary

Introduction

Bioaugmentation aims to use the capacities of specific bacterial strains inoculated into sites to enhance pollutant biodegradation. Whereas the questions of controlled invasion are old [15], we believe one can learn more about the process and its limitations from the bacteria themselves, how they perceive the transition from sterile culture medium into non-sterile contaminated sites, and which factors they express during growth and maintenance. Such knowledge may help to define specific process conditions favoring controlled growth within existing communities, and increase future success of bioaugmentation

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