Transcriptome analysis of a springtail, Folsomia candida, reveals energy constraint and oxidative stress during petroleum hydrocarbon exposure.

Petroleum hydrocarbon (PHC) contamination in soil is ubiquitous and poses harmful consequences to many organisms. The toxicity of PHC-impacted soil is difficult to predict due to variations in mixture composition and the impacts of natural weathering processes. Hence, high-throughput methods to assess PHC-impacted soils is required to expedite land management decisions. Next-generation sequencing is a robust tool that allows researchers to investigate the effects of contaminants on the transcriptome of organisms and identify molecular biomarkers. In this study, the effects of PHCs on conventional endpoints (i.e., survival and reproduction) and gene expression rates of a model springtail species, Folsomia candida were investigated. Age-synchronized F. candida were exposed to ecologically-relevant concentrations of soils spiked with fresh crude oil to calculate the reproductive EC25 and EC50 values using conventional toxicity testing. Soils spiked to these concentrations were then used to evaluate effects on the F. candida transcriptome over a 7-day exposure period. RNA-seq analysis found 98 and 132 differentially expressed genes when compared to the control for the EC25 and EC50 treatment groups, respectively. The majority of up-regulated genes were related to xenobiotic biotransformation reactions and oxidative stress response, while down-regulated genes coded for carbohydrate and peptide metabolic processes. Promotion of the pentose phosphate pathway was also found. Results suggest that the decreased reproduction rates of F. candida exposed to PHCs is due to energy constraints caused by inhibition of carbohydrate metabolic processes and allocation of remaining energy to detoxify xenobiotics. These findings provide insights into the molecular effects in F. candida following exposure to crude oil for seven days and highlight their potential to be used as a high-throughput screening test for PHC-contaminated sites. Adverse molecular effects can be measured as early as 24 h following exposure, whereas conventional toxicity tests may require a minimum of four weeks.

Phytotoxicity of weathered petroleum hydrocarbons in soil to boreal plant species.

Canada has extensive petroleum hydrocarbon (PHC) contamination in northern areas and the boreal forest region from historical oil and gas activities. Since the 2013 standardization of boreal forest species for plant toxicity testing in Canada, there has been a need to build the primary literature of the toxicity of weathered PHCs to these species. A series of toxicity experiments were carried out using fine-grained (<0.005-0.425 mm) background (100 total mg/kg total PHCs) and weathered contaminated soil (11,900 mg/kg total PHCs) collected from a contaminated site in northern Ontario, Canada. The PHC mixture in the contaminated site soil was characterized through Canadian Council of Ministers of the Environment Fractions, as indicated by the number equivalent normal straight-chain hydrocarbons (nC). The soil was highly contaminated with Fraction 2 (>nC10 to nC16) at 4790 mg/kg and Fraction 3 (>nC16 to nC34) at 4960 mg/kg. Five plant species (Elymus trachycaulus, Achillea millefolium, Picea mariana, Salix bebbiana, and Alnus viridis) were grown from seed in 0%, 25%, 50%, 75%, and 100% relative contamination mixtures of the PHC-contaminated and background soil from the site over 2-6 weeks. All five species showed significant inhibition in shoot length, shoot weight, root length, and/or root weight (Kruskal-Wallis Tests: p < 0.05, df = 4.0). Measurements of 25% inhibitory concentrations (IC25) following PHC toxicity experiments revealed that S. bebbiana was most significantly impaired by the PHC-contaminated soil (410-990 mg/kg total PHCs), where it showed <35% germination. This study indicates that natural weathering of Fraction 2- and Fraction 3-concentrated soil did not eliminate phytotoxicity to boreal plant species. Furthermore, it builds on the limited existing literature for toxicity of PHCs on boreal plants and supports site remediation to existing Canadian provincial PHC guidelines.

Assessment of the toxicity of weathered petroleum hydrocarbon impacted soils to native plants from a site in the Canadian Subarctic.

Remedial guidelines for petroleum hydrocarbons (PHCs) in soil aid in the mitigation of risks to human health and the environmental. However, some remediation guidelines may overestimate the potential for adverse effects to native plant species, contributing to unnecessary remedial efforts in attempts to meet the guidelines. At sites where PHC-contaminated soils undergo weathering, some PHCs may persist but with decreased bioavailability to organisms. In this study, the toxicity of both coarse and fine-grained subarctic soils, contaminated with weathered PHCs were assessed using five native plant species (Picea mariana, Achillea millefolium, Alnus viridis, Elymus trachycaulus and Salix bebbiana). Soil toxicity tests were conducted in a growth chamber with parameters set to simulate the site's subarctic climate conditions. Reference toxicant tests using boric acid were conducted to provide confidence in the interpretation of the results for the PHC-contaminated soils, and also provide new information on the sensitivities of the four boreal species to boric acid. All plants exhibited reduced growth and germination rates as boric acid concentrations increased. Despite exceeding the Canada-wide standard guidelines for Fraction 3 PHCs, field-collected contaminated soils had no significant negative impacts on the growth (i.e., length, dry weight and emergence) of any of the plant species tested.