Impact of atrazine concentration on bioavailability and apparent isotope fractionation in Gram-negative Rhizobium sp. CX-Z

Compound-specific stable isotope analysis of micropollutants has become an established method for the qualitative and quantitative assessment of biodegradation in the field. However, many of environmental factors may have an influence on the observed isotope fractionation. Herein, we investigate the impact of substrate concentration on the observed enrichment factor derived from Rayleigh plot of batch laboratory experiments conducted to measure the atrazine carbon isotope fractionation of Rhizobium sp. CX-Z subjected to the different initial concentration level of atrazine. The Rayleigh plot (changes in bulk concentration vs. isotopic composition) derived from batch experiments shown divergence from the linear relation towards the end of degradation, confirming bioavailability of atrazine changed along with the decay of substrate concentration, consequently, influenced the isotope fractionation and lowered the observed enrichment factor. When microbial degradation is coupled to a mass transfer step limiting the bioavailability of substrate, the observed enrichment factor displays a dependence on initial atrazine concentration. Observed enrichment factors (ε) (absolute value) derived from the low concentration (i.e. 9.5 μM) are below 3.5‰ to the value of -5.4‰ determined at high bioavailability (membrane-free cells). The observed enrichment factor depended significantly on the atrazine concentration, indicating the concentration level and the bioavailability of a substrate in realistic environments should be considered during the assessment of microbial degradation or in situ bioremediation based on compound-specific stable isotope analysis (CSIA) method.