Toxicological Response of Chironomus dilutus in Single-Chemical and Binary Mixture Exposure Experiments with 6 Perfluoralkyl Substances

Christopher J. McCarthy, Shaun A. Roark, Demitria Wright, Kelly O'Neal, Brett Muckey, Mike Stanaway, Justin N. Rewerts, Jennifer A. Field, Todd A. Anderson, Christopher J. Salice

Research output: Contribution to journalArticlepeer-review


Few studies have determined the toxicity of perfluoralkyl substances (PFAS) to aquatic invertebrates. We exposed Chironomus dilutus to 6 different PFAS to assess single-chemical toxicity and relative or proportional toxicity among substances. A 10-d range-finding test was conducted to inform 20-d assays for the following PFAS: perfluorooctane sulfonate (PFOS), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorobutanesulfonic acid (PFBS), perfluorohexanesulfonic acid (PFHxS), and perfluoroheptanoic acid (PFHpA). A 20-d binary mixture study of PFOS+PFHxS followed the single-chemical tests. Measurement endpoints for 20-d tests included larval survival and biomass. Log-logistic concentration response models were used to estimate 10, 20, and 50% effect concentrations (EC20, EC50) for PFOS, PFHxS, and PFOA. Survival EC50s for PFOS, PFHxS, and PFOA were 2.49, 3860, and 192 000 µg/L, respectively, whereas survival EC20s were 1.70, 913, and 119 000 µg/L for PFOS, PFHxS, and PFOA, respectively. Biomass as a combined survival and growth endpoint resulted in EC20s of 1.89, 896, and 137 000 µg/L for PFOS, PFHxS, and PFOA, respectively. Maximum concentrations tested (no-observed-effect concentrations) for PFNA, PFBS, and PFHpA were 2 to 3 orders of magnitude greater than the PFOS EC50s and showed no toxicity to C. dilutus, even at exposure concentrations well above what would be considered environmentally relevant. The binary mixture of 2.5 µg/L PFOS+1000 µg/L PFHxS showed reduced survival compared to controls and some indication of potential additive or synergistic interaction between PFOS and PFHxS. Overall, the present study supports previous studies showing PFOS to be the most toxic PFAS to aquatic life and suggests that PFOS could be more toxic to the freshwater midge than previously reported. Environ Toxicol Chem 2021;40:2319–2333.

Original languageEnglish
Pages (from-to)2319-2333
Number of pages15
JournalEnvironmental Toxicology and Chemistry
Issue number8
StatePublished - Aug 2021


  • Aquatic toxicity
  • Aqueous film–forming foam
  • PFAS mixture
  • Perfluoroalkyl substances (PFAS)
  • Perfluorohexanesulfonic acid
  • Perfluorooctane sulfonate


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