Biodegradation of chemicals tested in mixtures and individually: mixture effects on biodegradation kinetics and microbial composition

Biodegradation in the aquatic environment occurs in the presence of many chemicals, while standard simulation biodegradation tests are conducted with single chemicals. This study aimed to investigate the effect of the presence of additional chemicals on (1) biodegradation kinetics of individual chemicals and (2) the microbial composition in test systems. Parallel mixture and single substance experiments were conducted for 9 chemicals (phenethyl benzoate, oxacycloheptadec-10-en-2-one, alpha-ionone, methyl 2-naphthyl ether, decan-5-olide, octan-2-one, 2'-acetonaphthanone, methyl N-methylanthranilate, (+)-menthone) using inoculum from a Danish stream. Biotic and abiotic test systems were incubated at 12 degrees C for 1-30 days. Primary biodegradation kinetics were then determined from biotic/abiotic peak area ratios using SPME GC/MS analysis. The effect of the mixture on biodegradation varied with test chemical and was more pronounced for chemicals with lag-phases above 14 days: two chemicals degraded in the mixture but not when tested alone (i.e., positive mixture effect), and two degraded when tested alone but not in the mixture (i.e., negative mixture effect). Microbial composition (16S rRNA gene amplicon sequencing) was highly affected by 14 days incubation and the presence of the mixture (significant carbon source), but less by single chemicals (low carbon source). Growth on chemical mixtures resulted in consistent proliferation of Pseudomonas and Malikia, while specific chemicals increased the abundance of putative degraders belonging to Novosphingobium and Zoogloea. The chemical and microbiological results support (1) that simulation biodegradation kinetics should be determined in mixtures at low environmentally relevant concentrations and (2) that degradation times beyond some weeks are associated with more uncertainty. [GRAPHICS] .

Automated summary

This study investigated the effect of additional chemicals on the biodegradation kinetics and microbial composition in aquatic environments. It found that the impact of mixtures on degradation varied depending on the chemical, with longer lag-phases being more susceptible to mixture effects. Microbial composition was significantly affected by the presence of mixtures after 14 days of incubation, while individual chemicals had a lesser impact. Growth on chemical mixtures resulted in the proliferation of specific microorganisms, such as Pseudomonas and Malikia. These findings suggest that biodegradation kinetics should be determined in mixtures at environmentally relevant concentrations, and longer degradation times are associated with increased uncertainty.