Threshold for increased liver weight is protective of other effects in peromyscus exposed to PFNA.

Perfluorononanoic acid (PFNA) is a commercially relevant, long-chain (8 fully fluorinated carbon) perfluorinated carboxylic acid (PFCA). PFNA has limited terrestrial ecotoxicity data and is detected in humans, animals, and the environment. This study is the fourth in a series with the objective of investigating the toxicity of a suite of per- and polyfluoroalkyl substances (PFAS) detected on military installations in a mammal indigenous to North America. Peromyscus leucopus (white-footed mice, ∼25/sex/dose) were exposed via oral gavage to either 0, 0.03, 0.14, 1, or 3 mg PFNA/kg-day for 112 consecutive days (4 weeks pre-mating exposure followed by an additional 12 weeks of exposure after onset of mating). Parental generation animals were assessed for potential reproductive and developmental effects, organ weight changes, thyroid modulation, and immunotoxicity. Pup weight and survival were assessed at postnatal days 0, 1, 4, 7, and 10. Change in liver weight was determined to yield the most sensitive dose response according to benchmark dose analysis, and serves as the most protective point of departure (BMDL = 0.37 mg/kg-d PFNA). Other effects of PFNA exposure included reduced formation of plaque forming cells, which are indicative of functional immune deficits (BMDL = 2.31 mg/kg-d); decreased serum thyroxine (BMDL = 0.93 mg/kg-d) without changes in some other hormones; and increased stillbirths (BMDL = 0.61 mg/kg-d PFNA). Pup weight and survival were not affected by PFNA exposure. Combined with data from previous studies, data from Peromyscus provide a One Health perspective on health effects of PFAS.

Comparative Toxicity of Seven Aqueous Film-Forming Foam to In Vitro Systems and Mus.

The comparative toxicity of six per- and polyfluoroalkyl substance (PFAS)-free and one PFAS-containing aqueous film-forming foam (AFFF) was evaluated in an outbred mouse species as well as several in vitro assays. The in vivo toxicological profile of PFAS-free AFFFs in short-term, high-concentration exposures is different than that of a PFAS-containing AFFF. The PFAS-containing reference product induced increased liver weights, while the PFAS-free AFFFs were linked to either decreased or unaffected relative liver weights. The in vitro toxicological profile across PFAS-free AFFFs was uniform except in the Microtox® assay, where thresholds were variable and spanned several orders of magnitude. This direct comparison of products through short-term toxicity tests and in vitro screenings represents early data to support evaluation of potential regrettable substitutions when selecting alternative PFAS-free AFFFs. Further work in diverse taxa (e.g., aquatic organisms, terrestrial invertebrates, birds) and mammalian studies capturing sensitive life stages will refine and expand this data set across a range of risk-relevant toxicological endpoints. Environ Toxicol Chem 2023;42:2364-2374. Published 2023. This article is a U.S. Government work and is in the public domain in the USA.

Reproductive and immune effects emerge at similar thresholds of PFHxS in deer mice.

Although perfluorohexane sulfonate (PFHxS) is structurally similar to perfluorooctane sulfonate (PFOS) and also widely detected in humans and the environment, comparatively fewer toxicity data exists on this 6-chain perfluoroalkyl sulfonic acid. In this study, repeated oral doses of PFHxS were administered to deer mice (Peromyscus maniculatus) to evaluate subchronic toxicity and potential effects on reproduction and development. Maternal oral exposure to PFHxS caused increased stillbirths, which is relevant for ecological risk assessment, and resulted in a benchmark dose lower limit (BMDL) of 5.72 mg/kg-d PFHxS. Decreased plaque formation, which is relevant for human health risk assessment, occurred in both sexes of adult animals (BMDL = 8.79 mg/kg-d PFHxS). These data are the first to suggest a direct link between PFHxS and decreased functional immunity in an animal model. Additionally, female animals exhibited increased liver:body weight and animals of both sexes exhibited decreased serum thyroxine (T4) levels. Notably, since reproductive effects were used to support 2016 draft health advisories and immune effects were used in 2022 drinking water health advisories released by the United States Environmental Protection Agency for PFOS and perfluorooctanoic acid (PFOA), these novel data can potentially support advisories for PFHxS because relevant points of departure emerge at similar thresholds in a wild mammal and corroborate the general understanding of per- and polyfluoroalkyl substances (PFAS).

Toxicology assessment for six per- and polyfluoroalkyl (PFAS)-free aqueous film forming foam (AFFF) products.

There is an urgent need to understand the toxicity hazards of aqueous film forming foam (AFFF) replacement products to ensure the balance between performance and toxicity hazards and avoid regrettable substitutions during the rapid phasing out of per- and polyfluoroalkyl substance (PFAS)-containing AFFFs. To address this need, we assessed the toxicity of six candidate PFAS-free products via literature review, estimation techniques, and incorporation of testing data from whole products and compared them against one PFAS-containing product. Then, we combined the relative hazards across human occupational exposure (e.g., concentrate, foam, or dilute exposures), human environmental exposure (e.g., training, emergency response, cleanup), and environmental exposure to aquatic, mammalian, and other terrestrial species using an index-based scoring system to quantify potential hazards across these domains. We found that most PFAS-free products in their concentrated form may cause dermal and ocular irritation, and aquatic toxicity may be a concern from direct or repeated environmental releases. Additionally, all PFAS-free AFFF products assessed contain chemicals that are notable as plausible hazards resulting from release uncertainties (e.g., concentration, release volume, release timing), but the PFAS-free AFFF products appear to have a lower likelihood of environmental persistence and bioaccumulation and to have lower oral human health toxicity than the PFAS-containing reference product. Decision makers can use this information alongside cost-benefit, sustainability, or life-cycle analyses to make a data-driven decision for the adoption of PFAS-free AFFF. Integr Environ Assess Manag 2023;19:1609-1618. © 2023 SETAC.

Next-generation PFAS 6:2 fluorotelomer sulfonate reduces plaque formation in exposed white-footed mice.

6:2 fluorotelomer sulfonate (6:2 FTS) has been used as a replacement for legacy per- and polyfluoroalkyl substances (PFAS). We assessed reproductive and developmental effects in a human-wildlife hybrid animal model based on the association of adverse effects linked to legacy PFAS with these sensitive life stages. In this study, white-footed mice were exposed orally to 0, 0.2, 1, 5, or 25 mg/kg-day 6:2 FTS for 112 days (4 weeks premating exposure plus at least 4 weeks mating exposure). Pregnancy and fertility indices were calculated, and litter production, total litter size, live litter size, stillbirths, litter loss, average pup weight, and pinna unfolding were assessed. Sex steroid and thyroid hormone serum levels were assessed. Body weight, histopathology, and immune function were also assessed in this study. Reproductive endpoints were not significantly altered in response to 6:2 FTS. Spleen weight increased in male mice dosed with 6:2 FTS. Immune function determined via a plaque-forming cell (PFC) assay was decreased in both male and female mice in the 2 highest doses. A low benchmark dose was calculated based on PFCs as the critical effect and was found to be 2.63 and 2.26 mg/kg-day 6:2 FTS in male and female mice, respectively. This study characterizes 6:2 FTS as being potentially immunotoxic with little evidence of effect on reproduction and development; furthermore, it models acceptable levels of exposure. These 2 pieces of information together will aid regulators in setting environmental exposure limits for this PFAS currently thought to be less toxic than other PFAS.

Reproductive and developmental toxicity of perfluorooctane sulfonate (PFOS) in the white-footed mouse (Peromyscus leucopus).

Concerns about per- and polyfluoroalkyl substances (PFAS) stem from their ubiquitous presence in the environment, bioaccumulation, resistance to degradation, and toxicity. Previously, toxicity data relevant to ecological risk assessment has largely been aquatic, terrestrial invertebrates, or avian in origin. In this study, repeated oral exposures of perfluorooctane sulfonate (PFOS) were administered to white-footed mice (Peromyscus leucopus) to evaluate effects on reproduction and development. Prenatal exposure to high doses of PFOS caused neonatal mortality, though growth and development were unaffected by low doses. Additionally, parental (P) generation animals exhibited increased liver:body weight, increased hepatocyte cytoplasmic vacuolization, and decreased serum thyroxine (T4) levels. Total litter loss was selected as the protective critical effect in this study resulting in a benchmark dose low (BMDL) of 0.12 mg/kg-d PFOS. Importantly, PFOS exposure has been linked to reduced adult recruitment in myriad species and at similar thresholds to this study. Similarities in critical/toxicologic effects across taxa may add confidence in risk assessments at sites with multiple taxa or environments.

Comparative Toxicity of Herbicide Active Ingredients, Safener Additives, and Commercial Formulations to the Nontarget Alga Raphidocelis Subcapitata.

Chloroacetanilide herbicides are used worldwide to control weeds that affect crops such as corn, soybeans, and cotton. These herbicides are frequently paired with a "safener," which prevents herbicidal damage to the crop without diminishing weed control. Formulated herbicide products that include safeners and other ingredients are infrequently assessed for toxicity. Our goal was to understand the potential toxicity of safeners and herbicide + safener formulations relative to the toxicity of associated active ingredients. We quantified the concentration of safeners in commercially available formulations and tested effects on nontarget algae, Raphidocelis subcapitata, when exposed to individual herbicide active ingredients, safeners, and commercial formulations. The median effective concentrations (EC50s) causing 50% reduction in population growth for the herbicide active ingredients S-metolachlor and acetochlor were 0.046 and 0.003 ppm, respectively. The safeners benoxacor, AD-67, furilazole, and dichlormid were all substantially less toxic than the herbicides and were not toxic at environmentally relevant concentrations. The commercial formulations Dual II Magnum®, Me-Too-Lachlor II®, Harness®, and Surpass EC® all resulted in EC50 values that fell within the 95% confidence interval of the associated active ingredient herbicide. Interestingly, a significant increase in cell size was observed when algae were exposed to all the formulations, herbicides (acetochlor and S-metolachlor), and safener (dichlormid). The safener furilazole caused a significant decrease in cell size, whereas benoxacor and AD-67 had no observed effect on algae cell size. Significant algae cell size effects all occurred at or above the EC50 concentrations for each chemical, suggesting that other morphological effects may be occurring. Importantly, safeners in commercial formulations appeared not to impact toxicity to R. subcapitata compared with the active ingredient alone. Environ Toxicol Chem 2022;41:1466-1476. © 2022 SETAC.

Data of texture profile analysis performed by different input settings on stored 'Nui' and 'Rahi' blueberries.

Texture Profile Analysis is a well-established method for assessing mechanical properties of horticultural food products and consists of two compression cycles on a repeated motion to a given strain using a flat surface probe (i.e., compression plate). Input settings of target deformation (strain%) and duration (s) between compression cycles utilized for Texture Profile Analysis could influence output mechanical properties. The article provides data related to the ability of different Texture Profile Analysis operational settings to enable the separation of blueberries with variable mechanical properties. To create variable mechanical parameters of 'Nui' and 'Rahi' blueberries, fruit was stored in four relative humidity for 21 d at 4°C. For each storage humidity, mechanical properties of hardness (BH, N), hardness slope (BHS, kN m-1), apparent modulus of elasticity (E, MPa), and resilience (BR, -) were determined by utilizing two strain (15% or 30% of berry equatorial height). Meanwhile, mechanical parameters of cohesiveness (BCo, -), and springiness (BSp, -) were obtained by utilizing the combination of two strain (15% or 30%) and two duration between cycles (2 s and 10 s) as TPA operational settings. The statistical evaluation was conducted by one-way ANOVA, and the means of each storage humidity were separated according to the Tukey-HSD test (P = 0.05). The data presented in this article was used to select the Texture Profile Analysis operational settings utilized in the article entitled "Influence of water loss on mechanical properties of stored blueberries" Rivera et al. [1].

Patterns in Serum Toxicokinetics in Peromyscus Exposed to Per- and Polyfluoroalkyl Substances.

Per- and polyfluoroalkyl substances (PFAS) are compounds manufactured for use in paints, cleaning agents, fire suppressants, nonstick cookware, food containers, and water-resistant products. Concerns about PFAS stem from their ubiquitous presence in the environment, persistence, and variable/uncertain bioaccumulation and toxicity. In the present study, 5 perfluoroalkyl acids and one polyfluoroalkyl substance were administered to white-footed mice (Peromyscus leucopus) to elucidate the kinetics of each chemical over 28 d of exposure. Perfluorooctanoate, perfluorohexane sulfonate (PFHxS), and perfluorobutane sulfonate were administered to male and female mice via drinking water. Perfluorooctane sulfonate, perfluorononanoate, 6:2 fluorotelomer sulfonate, and PFHxS were administered to male and female mice via oral gavage. Blood samples collected after 14 or 21 and 28 d of exposure were analyzed for individual PFAS concentrations via liquid chromatography-tandem mass spectrometry. In general, a plateau in serum concentration in this toxicity test-relevant timeline depended on interactions between 1) the type of PFAS (i.e., perfluoroalkyl sulfonic acids [PFSAs] vs perfluoroalkyl carboxylic acids [PFCAs] vs polyfluorinated), 2) continuous versus bolus dosing, and 3) to a lesser extent, sex. Specifically, PFCAs were detected at higher concentration in females than males, whereas PFSAs were generally detected at similar levels across sex. An exception occurred when PFHxS yielded higher serum levels in males than females through bolus, but not continuous, dosing. Type of PFAS had the largest impact on serum concentrations, whereas sex had the lowest. As such, future work on the toxicokinetics of PFAS in common ecological receptors would be valuable to further explore these patterns. Environ Toxicol Chem 2021;40:2886-2898. © 2021 SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Application of Absorption and Scattering Properties Obtained through Image Pre-Classification Method Using a Laser Backscattering Imaging System to Detect Kiwifruit Chilling Injury.

Kiwifruit chilling injury (CI) damage occurs after long-term exposure to low temperature. A non-destructive approach to detect CI injury was tested in the present study, using a laser backscattering image (LBI) technique calibrated with 56 liquid phantoms for providing absorption coefficient (µa) and reduced scattering coefficient (µs'). Calibration of LBI resulted in a true-positive (TP) classification of 91.5% and 65.6% of predicted µs' and µa, respectively. The optical properties of 'SunGold™'and 'Hayward' kiwifruit were analysed at 520 nm with a two-step protocol capturing pre-classification according to the LBI parameters used in the calibration and estimation with the Farrell equation. Severely injured kiwifruit showed white corky tissue and water soaking, reduced soluble solids content and firmness measured destructively. Non-destructive classification results for 'SunGold™' showed a high percentage of TP for severe CI of 92% and 75% using LBI parameters directly and predicted µa and µs' after pre-classification, respectively. The classification accuracy for severe CI 'Hayward' kiwifruit with LBI parameter was low (58%) and with µa and µs' decreased further (35%), which was assumed to be due to interference caused by the long trichomes on the fruit surface.

Per- and Polyfluoroalkyl Substances (PFAS) in Surface Water Near US Air Force Bases: Prioritizing Individual Chemicals and Mixtures for Toxicity Testing and Risk Assessment.

Per- and polyfluoroalkyl substances (PFAS) are a large class of persistent chemicals used for decades in industrial and commercial applications. A key challenge with regard to estimating potential risk to ecological (and human) receptors associated with PFAS exposure lies in the fact that there are many different PFAS compounds and several to many can co-occur in any given environmental sample. We applied a data science approach to characterize and prioritize PFAS and PFAS mixtures from a large dataset of PFAS measurements in surface waters associated with US Air Force Installations with a history of the use of aqueous film-forming foams (AFFFs). Several iterations of stakeholder feedback culminated in a few main points that advanced our understanding of a complex dataset and the larger ecotoxicological problem. First, perfluorooctane sulfonate (PFOS) was often a dominant PFAS in a given surface water sample, frequently followed by perfluorohexane sulfonate (PFHxS). Second, a 4-chemical mixture generally accounted for >80% of the sum of all routinely reported PFAS in a sample, and the most representative 4-chemical mixture was composed of PFOS, PFHxS, perfluorohexanoic acid (PFHxA), and perfluorooctanoic acid (PFOA). We suggest that these results demonstrate the utility of formalized data science analysis and assessment frameworks to address complex ecotoxicological problems. Specifically, our example dataset results can be used to provide perspective on toxicity testing, ecological risk assessments, and field studies of PFAS in and around AFFF-impacted sites. Environ Toxicol Chem 2021;40:871-882. © 2020 SETAC.

Intraspecific interactions affect outcomes of pulse toxicity at different Daphnia magna population phases.

Traditional toxicity tests assess stressor effects on individuals, while protection goals are focused on the population-level and above. Additionally, these tests ignore common ecological factors such as resource levels and population growth phase. The objective of this research was to explore effects of - and interactions between - resource availability and stress response at the individual and population levels using Daphnia magna as a model. We hypothesized that density-dependent changes in resources at various phases of population growth would cause different population responses to the same toxicant stress. Laboratory populations of Daphnia magna were exposed to a 48-h pulse of 20 or 30 µg/l pyraclostrobin in one of four distinct phases of laboratory population cycles: growth, peak, decline, and stable. Population size and recovery were observed throughout the 51-day study. Populations exposed to pyraclostrobin during the growth phase had the least mortality and fastest recovery, while populations in the peak phase had the greatest mortality and slowest recovery. These data suggested that high density and low food at the peak phase resulted in more sensitive daphnids. To further test this hypothesis, a resource-amended acute toxicity study was conducted to quantify the effects of food resource on pyraclostrobin toxicity to Daphnia magna. Three age classes of Daphnia magna (neonate, subadult, adult) were fed low or high food levels and exposed to pyraclostrobin for 48 h. Toxicity was greater, as shown by lower 48 h LC50s, for smaller Daphnia magna age classes and lower food levels comporting results in the population study. Importantly, the acute toxicity studies generally yielded lower effect levels than the population studies suggesting that while the standard acute studies are ecologically unrealistic, they may be protective of toxicity under some circumstances. Collectively, these data point to the importance of population phase and the resource environment in modulating toxicity.

Insights Into Olive Fruit Surface Functions: A Comparison of Cuticular Composition, Water Permeability, and Surface Topography in Nine Cultivars During Maturation.

Olive (Olea europaea L.) growing has outstanding economic relevance in Spain, the main olive oil producer and exporter in the world. Fruit skin properties are very relevant for fruit and oil quality, water loss, and susceptibility to mechanical damage, rots, and infestations, but limited research focus has been placed on the cuticle of intact olive fruit. In this work, fruit samples from nine olive cultivars ("Arbequina," "Argudell," "Empeltre," "Farga," "Manzanilla," "Marfil," "Morrut," "Picual," and "Sevillenca") were harvested from an experimental orchard at three different ripening stages (green, turning, and ripe), and cuticular membranes were enzymatically isolated from fruit skin. The total contents of cuticular wax and cutin significantly differed among cultivars both in absolute and in relative terms. The wax to cutin ratio generally decreased along fruit maturation, with the exception of "Marfil" and "Picual." In contrast, increased water permeance values in ripe fruit were observed uniquely for "Argudell," "Morrut," and "Marfil" fruit. The toluidine blue test revealed surface discontinuities on green samples of "Argudell," "Empeltre," "Manzanilla," "Marfil," and "Sevillenca" fruit, but not on "Arbequina," "Farga," "Morrut," or "Picual." No apparent relationship was found between water permeability and total wax coverage or the results of the toluidine blue test. The composition of cuticular waxes and cutin monomers was analyzed in detail, and sections of fruit pericarp were stained in Sudan IV for microscopy observations. Skin surface topography was also studied by means of fringe projection, showing large differences in surface roughness among the cultivars, "Farga" and "Morrut" fruits displaying the most irregular surfaces. Cultivar-related differences in cuticle and surface features of fruit are presented and discussed.

Will temperature increases associated with climate change potentiate toxicity of environmentally relevant concentrations of chloride on larval green frogs (Lithobates clamitans)?

An important challenge in amphibian ecotoxicology and conservation is that amphibian toxicity tests are usually focused on a single chemical while populations experience multiple, simultaneous stressors. For example, about 14 million tons of road de-icing salts are used each year in North America with NaCl accounting for 98% of total salt use and, hence, elevated chloride is an important environmental stressor to aquatic organisms, including amphibians. As well, higher temperature as a result of climate change is becoming an increasingly important environmental stressor. There are no data on the combined effects of chloride and temperature on amphibians hinders conservation efforts. We conducted field studies to characterize chloride concentrations and water temperatures in known amphibian breeding habitats and performed toxicity tests to explore impacts of these two stressors on a common anuran, the green frog (Lithobates clamitans). A 96-hour acute toxicity test was conducted to first determine a chloride LC50 (2587.5 mg Cl-/L) at a single, neutral temperature, which was used to inform the treatment levels of the sub-chronic test, which also included a temperature range. In the sub-chronic study, green frog larvae were exposed to three temperatures (18, 22, and 25 °C), and four concentrations of chloride (0, 500, 1000, and 2000 mg Cl-/L) for 35 days. At all temperatures, tadpoles exposed to 2000 mg Cl-/L had significantly higher mortality. While there was no significant effect of temperature alone on mortality, survival of tadpoles was significantly lower at 1000 mg Cl-/L at the two higher temperatures suggesting a potentiation of chloride ion toxicity with increasing temperature. Comparing toxicity results to field measurements of chloride and temperature suggests green frog tadpoles and other species with similar sensitivity are likely negatively affected. Data on additional species and populations would further increase our understanding of how salt and temperature may shape aquatic communities.

Measuring ethylene in postharvest biology research using the laser-based ETD-300 ethylene detector.

BACKGROUND: Ability to measure ethylene is an important aspect of postharvest management, as knowledge of endogenous ethylene production is used in assessing physiological status, while response of crops to exogenous ethylene informs efforts needed to control unwanted ripening. An ethylene monitoring device with a laser-based photoacoustic detector, ETD-300, was recently developed by Sensor Sense B.V., Nijmegen, The Netherlands. In terms of performance, the ETD-300 is superior to all other current ethylene measurement devices, with a sensitivity of 0.3 nL L-1, a response time of 5 s, and an ability to monitor ethylene in real time. Although the ETD-300 is relatively easy to operate, the performance and correctness of the data obtained depends on the choice of settings, which depends on the application. RESULTS: This article provides a description of different ways in which the ETD-300 can be used in postharvest research for monitoring ethylene production and ethylene presence in an environment. We provided guidelines on selecting the appropriate method (Continuous Flow, Stop and Flow, and Sample methods), and operational curves for deciding on suitable combination of free volume, flow rates, and period for the different measurement methods. CONCLUSIONS: Using these guidelines and operational curves, ETD-300 users can considerably reduce the measurement effort by limiting trial and error in establishing appropriate methodologies for their application. The guidelines also comment on accurate use of the ETD-300, as using the inappropriate settings could lead to erroneous measurements. Although these methodologies were developed primarily for postharvest application, they can be applied in other plant science research.