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Per- and poly-fluoroalkyl substances (PFAS) pose a threat to organisms and ecosystems due to their persistent nature. Ecotoxicology endpoints used in regulatory guidelines may not reflect multiple, low-level but persistent stressors. This study examines the biological effects of PFAS on Eastern short-necked turtles in Queensland, Australia. In this study, blood samples were collected and analysed for PFAS, hormone levels, and functional omics endpoints. High levels of PFAS were found in turtles at the impacted site, with PFOS being the dominant constituent. The PFAS profiles of males and females differed, with males having higher PFAS concentrations. Hormone concentrations differed between impacted and reference sites in male turtles, with elevated testosterone and corticosterone indicative of stress. Further, energy utilisation, nucleotide synthesis, nitrogen metabolism, and amino acid synthesis were altered in both male and female turtles from PFAS-impacted sites. Both sexes show similar metabolic responses to environmental stressors from the PFAS-contaminated site, which may adversely affect their reproductive fitness. Purine metabolism, caffeine metabolism, and ferroptosis pathway changes in turtles can cause gout, cell death, and overall health problems. Further, the study showed that prolonged exposure to elevated PFAS levels in the wild could compromise turtle reproductive fitness by disrupting reproductive steroids and metabolic pathways., Competing Interests: Declaration of competing interest The authors declare that they have no known competing commercial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Turtles are a potential sentinel species of aquatic ecosystem health as they inhabit aquatic ecosystems, are long lived, and potentially have high exposure to anthropogenic chemicals via food and water. This study investigated per- and polyfluoroalkyl substances (PFAS) tissue partitioning in female Emydura macquarii macquarii turtle, and the maternal offloading of (PFAS) into eggs and then hatchlings as well as the accumulation of PFAS in male and female Emydura macquarii macquarii serum. Significantly higher levels of perfluorosulfonic acids (PFSAs) and perfluorocarboxylic acids (PFCAs) were measured in the male serum compared to the female turtle serum, whereas perfluoroalkane sulfonamides (FASAs) were significantly higher in the female turtle serum. Perfluorooctane sulfonate (PFOS) was the predominant PFAS in the turtles whereas PFHxA was the predominant PFAS found in the surrounding water. PFHxA was not reported in any turtle tissue or the serum. The short-chain PFSAs and FASAs appeared to be highly associated with blood; long-chain PFSAs and PFCAs were more likely to be associated with tissue. Half of the PFHxS and all the long-chain PFSAs and PFCAs reported in the yolks were transferred into the hatchlings (by mass), suggesting a potential intergenerational effect., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2024. Published by Elsevier B.V. All rights reserved.)

Background: Per- and polyfluoroalkyl substances (PFAS) have been associated with higher cholesterol and liver function markers in some studies, but the evidence for specific cardiometabolic conditions has been inconclusive., Objectives: We quantified the associations of single and combined PFAS with cardiometabolic markers and conditions in a cross-sectional study of three Australian communities with PFAS-contaminated water from the historical use of aqueous film-forming foam in firefighting activities, and three comparison communities., Methods: Participants gave blood samples for measurement of nine PFAS, four lipids, six liver function markers, and completed a survey on sociodemographic characteristics and eight cardiometabolic conditions. We estimated differences in mean biomarker concentrations per doubling in single PFAS concentrations (linear regression) and per interquartile range increase in the PFAS mixture (Bayesian kernel machine regression). We estimated prevalence ratios of biomarker concentrations outside reference limits and self-reported cardiometabolic conditions (Poisson regression)., Results: We recruited 881 adults in exposed communities and 801 in comparison communities. We observed higher mean total cholesterol with higher single and mixture PFAS concentrations in blood serum (e.g., 0.18 mmol/L, 95% credible interval -0.06 to 0.42, higher total cholesterol concentrations with an interquartile range increase in all PFAS concentrations in Williamtown, New South Wales), with varying certainty across communities and PFAS. There was less consistency in direction of associations for liver function markers. Serum perfluorooctanoic acid (PFOA) concentrations were positively associated with the prevalence of self-reported hypercholesterolemia in one of three communities, but PFAS concentrations were not associated with self-reported type II diabetes, liver disease, or cardiovascular disease., Discussion: Our study is one of few that has simultaneously quantified the associations of blood PFAS concentrations with multiple biomarkers and cardiometabolic conditions in multiple communities. Our findings for total cholesterol were consistent with previous studies; however, substantial uncertainty in our estimates and the cross-sectional design limit causal inference., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Professor Martyn Kirk worked part-time for the Australian Government Department of Health between 2020 and 2022 on the Australian national COVID-19 response., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Human biomonitoring programs of per- and polyfluoroalkyl substances (PFAS) have been conducted around the world to assess human exposure and health risk. Inquiry into population PFAS levels in a socioeconomically and geographically unique region such as the Pacific Island Papua New Guinea, may provide new insights into PFAS exposure pathways and sources. This study presented the first indication of PFAS exposure in the Papua New Guinea population. De-identified serum samples were pooled from surplus pathology serum samples collected between 2019 and 2020. A total of 11 PFAS were detected in the serum pools including 10 perfluoroalkyl acids (PFAA) and 9Cl-F53B (a perfluorooctane sulfonic acid - PFOS alternative). The observed PFAA profile was somewhat similar to that observed for general population data of other countries such as Australia, Malaysia, and Canada suggesting similar exposure sources and/or pathways. However, PFAS concentrations were consistently lower than concentrations in the serum measured in pools obtained from Australia. The detection of 9Cl-F53B in all pools was a new finding which might be related to exposure from locally industrial sources., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Elevated levels of perfluorooctane sulfonate (PFOS) and elevated detection frequency of chloro-substituted PFOS have been reported in Australian firefighters with historical exposure to aqueous-film forming foam (AFFF). The aim of this study is to estimate the apparent half-lives of Cl-PFOS and PFOS isomers in firefighters following the end of exposure to 3M-AFFF. Paired serum samples from 120 firefighters, collected approximately five years apart, were analyzed for 8-Cl-PFOS (8-chloroperfluoro-1-octanesulfonic acid) and PFOS isomers via targeted LC-MS/MS. Apparent half-life was estimated by assuming a first order-elimination model. Cl-PFOS was detected in 93% of all initial serum samples (

Objectives: Firefighters who used aqueous film forming foam in the past have experienced elevated exposures to perfluoroalkyl acids (PFAAs). The objective of this study was to examine the associations between clinical chemistry endpoints and serum concentrations of perfluorooctanoic acid (PFOA), perfluorohexane sulfonate (PFHxS), perfluoroheptane sulfonate (PFHpS) and perfluorooctane sulfonate (PFOS) in firefighters. Multiple linear regression was used to assess relationships between PFAA serum concentrations and biochemical markers for cardiovascular disease, kidney-, liver- and thyroid function, in a cross-sectional survey of 783 firefighters with elevated levels of PFHxS, PFHpS and PFOS in relation to the most recently reported levels in the general Australian population. Linear logistic regression was used to assess the odds ratios for selected self-reported health outcomes. Repeated measures linear mixed models were further used to assess relationships between PFAAs and biomarkers for cardiovascular disease and kidney function longitudinally in a subset of the firefighters (n = 130) where serum measurements were available from two timepoints, five years apart. In the cross-sectional analysis, higher levels of all PFAAs were significantly associated with higher levels of biomarkers for cardiovascular disease (total-cholesterol, and LDL-cholesterol). For example, doubling in PFOS serum concentration were associated with increases in total cholesterol (β:0.111, 95% confidence interval (95%CI): 0.026, 0.195 mmol/L) and LDL-cholesterol (β: 0.104, 95%CI:0.03, 0.178 mmol/L). Doubling in PFOA concentration, despite not being elevated in the study population, were additionally positively associated with kidney function marker urate (e.g., β: 0.010, 95%CI; 0.004, 0.016 mmol/L) and thyroid function marker TSH (e.g., β: 0.087, 95%CI: 0.014, 0.161 mIU/L). PFAAs were not associated with any assessed self-reported health conditions. No significant relationships were observed in the longitudinal analysis. Findings support previous studies, particularly on the association between PFAAs and serum lipids., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Firefighters may be occupationally exposed to per- and polyfluoroalkyl substances (PFASs) through Aqueous Film-Forming Foam (AFFF), smoke, dust and turnout gear, in addition to other background exposure sources. Epidemiological assessment of PFAS exposure in an occupational cohort of firefighting staff commenced in 2013-2014, following cessation of PFAS-based AFFF in Australian aviation. Here we present the study design and methodology of a follow-up study conducted in 2018-2019. We focus on our experiences engaging with stakeholders and participants with the establishment of an inclusive study group and highlight the key lessons learned from implementing a co-design process in the study. The study included a cross-sectional assessment of blood serum concentrations of 40 PFASs, including perfluorooctanoic acid (PFOA), perfluorooctane sulfonate (PFOS) and perfluorohexane sulfonate (PFHxS), and 14 health-related biomarkers in 799 current and former Aviation Rescue Firefighting Services employees. A large proportion (87%) of the participants from the preliminary exposure study in 2013-2014 were re-recruited in the follow-up study. This enabled further longitudinal analyses in this subset of 130 participants. Participants included employees from different work roles and timeframes, reflecting the periods when three different firefighting foams were utilised in Australia. Establishment of a collaborative and inclusive study group (including stakeholders and participants) contributed to several components of the study design, including the expansion of robust analytical quality assurance and control measurements, and tailoring of communication and dissemination strategies. These outcomes were key factors that improved transparency of the research design, methods and results. Additionally, implementing elements of co-design helped build trust between researchers and participants, which is an important consideration for studies funded by stakeholders related to the exposure source., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

PFAS mixtures in the environment are common and identifying PFAS constituents, bioaccumulation, and biological impacts of mixtures remains a challenge. Here, an omics-based ecosurveillance approach was taken to investigate the impacts of PFAS pollution in freshwater turtles (Emydura macquariimacquarii). Four turtles were collected from an impacted waterway downstream from an industrial source of PFAS contamination in Queensland, Australia and analysed for 49 different PFAS. One turtle was collected from a suitable control site. PFAS concentrations were quantified in turtle serum using an established targeted methodology. The serum PFAS concentration was ten-fold greater at the impacted site (Σ49 PFAS 1933 ± 481 ng/mL) relative to the control sample (Σ49 PFAS 140 ng/mL). Perfluorooctane sulfonate (PFOS; 889 ± 56 ng/mL) was 235 times higher in turtle serum than in the water that they were collected from (ΣPFAS 32.0 μg/L). Perfluorobutane sulfonamide (FBSA; 403 ± 83 ng/mL) and perfluorohexane sulfonamide (FHxSA; 550 ± 330 ng/mL) were also reported at substantial concentrations in the serum of impacted turtles. Biochemical profiles were analysed using a mixture of liquid chromatography triple quadrupole (QqQ) and quadrupole time-of-flight (QToF) mass spectrometry methodologies. These profiles demonstrated a positive correlation in the impacted turtles exposed to elevated PFAS with an enhanced purine metabolism, glycerophosphocholines and an innate immune response, which suggest an inflammation response, metabolic preservation and re-routing of central carbon metabolites. Conversely, lipid transport and binding activity were negatively correlated. Using these preliminary data, we were able to demonstrate the negative metabolic impact from PFAS mixtures on turtle metabolic health. With further research on a larger turtle cohort, omics-based data will contribute towards linking adverse outcome pathways for turtle populations exposed to PFAS mixtures. Moreover, expanding the use of ecosurveillance tools will inform mechanistic toxicological data for risk assessment and regulatory applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2021. Published by Elsevier B.V. All rights reserved.)

Per- and poly-fluoroalkyl substances (PFAS) are a range of persistent organofluorine contaminants, some of which have been found to accumulate in humans and have long half-lives. In longitudinal studies, when relying on measurements obtained at different points in time, it is critical to understand the associated analytical uncertainties when interpreting the data. In this manuscript we assess precision measurements of serum PFAS analysis in a follow-up study undertaken approximately 5 years after the initial study. These measurements included intra-(n = 58) and inter-batch duplicates (n = 57), inter-batch replicates (n = 58), inter-laboratory replicates (n = 10) and a re-analysis of 120 archived serum samples from the initial study. Average coefficients of variation (CV) for perfluorooctanoic acid (PFOA), perfluorohexane sulfonate (PFHxS) and perfluorooctane sulfonate (PFOS) associated with the reanalysis of archived samples ranged from 4 to 8%, which was greater than the inter- and intra -batch duplicates (<3%), but lower than the inter-laboratory comparison (CV ≥ 10%). Multi-centre analytical capacity in studies increases the variance within the dataset and implementation of variability-measures are useful to refine and maintain comparability. Due to long PFAS half-lives, this variance is an important consideration when deciding appropriate time intervals for sample collections in longitudinal studies, to ensure the difference is greater than the analytical uncertainty., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Previous studies have shown that accumulation of perfluoroalkyl acids (PFAAs) in the tissues of aquatic species is highly variable. Movement and migration patterns in these species represent an important consideration when evaluating contaminant accumulation in exposed biota, and may have a large influence on the risk profiles for migratory seafood species. In this study, relationships between PFAA concentrations in muscle and liver tissue, and recent fish migration history (inferred from metals profiles in fish otoliths, otherwise known as otolith chemistry) were evaluated in Sea Mullet (Mugil cephalus). A greater number of PFAAs, and higher concentrations, were found in liver compared to muscle tissue. Perfluorooctane sulfonate (PFOS) was present in highest concentrations in both muscle and liver tissues, and there was strong correlation in concentrations between these two tissues. PFOS was found to decrease and increase alongside recent strontium and barium concentrations (respectively) in the otolith, suggesting higher concentrations of PFAAs in fish recently exposed to comparatively lower salinity environments. This study highlights how otolith chemistry can be employed to examine links between contaminant concentrations in fish, and their recent migration history. This approach shows promise for studying contaminant residues in mobile seafood species within the natural environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2021. Published by Elsevier Ltd. All rights reserved.)

Poly and perfluorinated alkyl substances (PFASs) are persistent organic pollutants (POPs) that are highly resistant to environmental degradation, and have been detected in a broad range of terrestrial and aquatic species. Portunid crabs have been shown to accumulate comparatively high concentrations of PFASs, but previous work examining depuration in crabs was inconclusive. Here, we trialled a novel experimental design to study depuration of PFASs from edible tissues of portunid crabs, using paired claw samples, and trial this design with Giant Mud Crab Scylla serrata exposed to the contaminant under natural conditions. We found evidence for depuration of perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHxS) and perfluorooctanoic acid (PFOA), but with depuration half-lives as high as 40 days (for PFOS). We also observed substantial variability in the data, including differences in PFAS concentrations between claws from the same individuals, potentially resulting from claw loss and re-growth prior to capture. These results have broad implications for assessing and minimising exposure risk in seafood species., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2020. Published by Elsevier B.V. All rights reserved.)