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, α-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 °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.

Determining the Temperature Dependency of Biodegradation Kinetics for 34 Hydrocarbons while Avoiding Chemical and Microbial Confounding Factors.

Biodegradation kinetics data are keystone for evaluating the environmental persistence and risk of chemicals. Biodegradation kinetics depend highly on the prevailing temperature, which influences microbial community structures, metabolic rates, and chemical availability. There is a lack of high-quality comparative biodegradation kinetics data that are determined at different test temperatures but with the same microbial inoculum and chemical availability. The present study was designed to determine the effect of test temperature on the biodegradation kinetics of hydrocarbons while avoiding confounding factors. We used inocula from a Northern river (2.7 °C) and a Central European river (12.5 °C). Aqueous stock solutions containing 45 individual hydrocarbons were generated by passive dosing and added to river water containing the native microorganisms. Compound-specific biodegradation kinetics were then determined at 2.7, 12, and 20 °C based on substrate depletion. Main findings comprise the following: (1) Degradation half-times (DegT50) of 34 test chemicals were determined at different test temperatures and were largely consistent with the Arrhenius equation (activation energy, 65.4 kJ/mol). (2) Differences in biodegradation kinetics between tested isomers were rather limited. (3) The recent lowering of standard test temperature from 20 to 12 °C results typically in a doubling of DegT50 values and can lead to a stricter persistency assessment.

Combining Headspace Solid-Phase Microextraction with Internal Benchmarking to Determine the Elimination Kinetics of Hydrophobic UVCBs.

Substances classified as unknown or variable composition, complex reaction products or biological origin (UVCB) present a challenge for environmental hazard and risk assessment. Here, we present a novel approach for whole-substance bioconcentration testing applied to cedarwood oil-an essential oil composed of volatile, hydrophobic organic chemicals. The method yields whole-body elimination rate constants for a mixture of constituents. Our approach combines in vivo dietary fish exposure with internal benchmarking and headspace solid-phase microextraction (HS-SPME) equilibrium sampling followed by suspect-screening analysis. We quantified depuration rate constants of 13 individual cedarwood oil constituents based on relative peak areas using gas chromatography (GC) coupled with Orbitrap-mass spectrometry (MS) and GC triple-quadrupole (QqQ)-MS. For seven constituents with available analytical standards, we compared the rate constants to the results obtained from solvent extraction, clean-up, and targeted GC-MS analysis. The HS-SPME sampling approach allowed for automated sample extraction and analyte enrichment while minimizing evaporative losses of the volatile target analytes and reducing matrix interferences from low-volatility organics. The suspect-screening analysis enabled the quantification of constituents without available analytical standards, while the internal benchmarking significantly reduced variability from differences in delivered dose and analytical variability between the samples.

Accelerated equilibrium sampling of hydrophobic organic chemicals in solid matrices: A proof of concept on how to reach equilibrium for PCBs within 1 day.

Equilibrium sampling of hydrophobic organic chemicals (HOCs) is increasingly used to measure freely dissolved concentrations and chemical activities in sediments and soils. However, for the most hydrophobic chemicals (Log Kow > 6) such equilibrium sampling requires often very long sampling times in the order of weeks to months. The aim of the present study was to explore two strategies for markedly increasing the HOC mass transfer from matrix to sampler with the overall goal to shorten equilibration times down to a few hours. Two Solid Phase Microextraction (SPME) approaches were thus developed and tested in sediment and soil contaminated by polychlorinated biphenyls (PCBs). In the first method, the SPME fiber was immersed directly in the aqueous suspension of the sample under vigorous agitation. In the second method equilibration took place via the headspace and was accelerated by elevating the temperature. Headspace-SPME at 80 °C provided fast equilibration within approximately 2 h without contacting the sample and thus avoiding fiber fouling. Both SPME methods were calibrated by passive dosing from preloaded silicone rods and yielded similar results, supporting the validity of HS-SPME at elevated temperatures on a proof of principle level. Finally, by using 13C labelled PCB standards, total concentrations were simultaneously measured, which in turn allowed calculation of matrix-water distribution coefficients.

Thermodynamic assessment of (semi-)volatile hydrophobic organic chemicals in WWTP sludge - combining solid phase microextraction with non-target GC/MS.

Applying WWTP sludge on arable soil has clear benefits from a resource recycling point of view but can potentially also lead to contamination of soil, agricultural products and the environment. The sludge contains a complex mixture of particularly hydrophobic organic chemicals (HOCs) that sorb to the organic matter. Equilibrium sampling was recently applied to the measurement of chemical activities of polycyclic aromatic hydrocarbons (PAHs) in secondary and digested sludge, and clear activity increases due to the anaerobic digestion treatment were observed. In the present study we extend this work to a large number of (semi-)volatile HOCs by combining automated headspace solid phase microextraction with non-targeted gas chromatography mass spectrometry. Chemical activity ratios were determined between sludge from the different stages of a WWTP and after co-composting with garden waste and sorbent amendment with activated carbon (AC) and biochar (BC). Generally, chemical activities increased from primary, to secondary, to digested sludge and the level in the dewatered sludge was not significantly different from the level in the digested sludge. Decamethylcyclopentasiloxane (D5) behaved differently as the level was similar until the dewatering step, where it increased 4-fold. The results confirmed the earlier observation that anaerobic digestion increased chemical activity, now for a broader range of chemicals, and showed that co-composting was effective in reducing chemical activities of most of the tested (semi-)volatile organic chemicals. Of the studied compounds, activities of D5 and a musk fragrance were reduced the least by co-composting.