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Toxicological study of PFOS/PFOA to zebrafish (Danio rerio) embryos
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Acute toxicity of PFOS/PFOA to zebrafish (Danio rerio) and development effects to zebrafish embryo were examined using a zebrafish embryo test. PFOS/PFOA showed remarkably toxicity effects on zebrafish. The LC50 (48 h) values are 1 005 mg/L for PFOA, 107 mg/L for PFOS, while the LC50 (96 h) values are 499 mg/L for PFOA, 71 mg/L for PFOS. Moreover PFOS/PFOA inhibited embryo development, and caused embryo abnormality and death. After exposure to high concentration of PFOS (> 240 mg/L), cells in animal pole of embryos autolyzed to coagulate, which indicated PFOS caused cell membranes damage. The most sensitive endpoints for PFOS exposure is spinal column malformation, and the EC50 values is 9.14 mg/L. While for PFOA hatching (96 h) is the most sensitive, and the EC50 values is 328.0 mg/L. Both PFOS and PFOA retarded embryo development which indicates their development toxicity.
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... Lethal effects of PFOS on zebrafish have been listed in Table 1. PFOS can reduce embryo and larval survivorship in zebrafish (D. rerio) Du et al., 2009;Hu, 2009;Ye et al., 2009;Kim et al., 2011;Wang et al., 2011;Zheng et al., 2012;Li et al., 2015;Zhao et al., 2016). The 48-h, 96-h, and 120-h LC50 of PFOS to zebrafish embryos were 31.27-107 ...
... The 48-h, 96-h, and 120-h LC50 of PFOS to zebrafish embryos were 31.27-107 mg/L (Ye et al., 2009;Li et al., 2015), 10.89-71 mg/L (Ye et al., 2009;Li et al., 2015) and 2.2-28.21 mg/L (Huang et al., 2010;Hagenaars et al., 2011), respectively; while 72-h, 96-h and 120-h LC50 to larvae were 68 mg/L (Zheng et al., 2012), 31.03 ...
... The 48-h, 96-h, and 120-h LC50 of PFOS to zebrafish embryos were 31.27-107 mg/L (Ye et al., 2009;Li et al., 2015), 10.89-71 mg/L (Ye et al., 2009;Li et al., 2015) and 2.2-28.21 mg/L (Huang et al., 2010;Hagenaars et al., 2011), respectively; while 72-h, 96-h and 120-h LC50 to larvae were 68 mg/L (Zheng et al., 2012), 31.03 ...
Rapid industrial development and extensive use of chemicals have resulted in elevated concentrations of emerging contaminants worldwide, posing a substantial threat to the ecological environment and human health. Per- and polyfluoroalkyl substances (PFASs) have been recognized as emerging pollutants that are widely distributed and accumulated in the environment and they have drawn the attention of scholars for several decades. The variety, long-term use, and long-distance transmission of PFASs have resulted in the ubiquitous contamination of global ecosystems, especially in aquatic environments. Ever since perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) were added to the Stockholm Convention on Persistent Organic Pollutants (POPs), they have become the most typical, eye-catching, and frequently investigated PFASs. Owing to the high stability and bioaccumulation of PFASs, as well as the adverse impact on the endocrine, immune, and nervous systems, investigating their contamination levels, risk of transfer along the food chain, and ecotoxicity should be prioritized. In addition to the important evolutionary significance as primitive vertebrates and the main consumers of aquatic environment, fishes generally exist in various aquatic food chains from the bottom to the top and occupy a critical position in terms of aquatic ecology protection; while amphibians, as the key link from aquatic to terrestrial organisms, are highly sensitive to different environmental pollutants. This review is a comprehensive summary of the toxic effects and toxicity-related factors of PFASs on aquatic vertebrates, mainly Pisces and Amphilla organisms, the characteristics of different aquatic vertebrates in toxicity investigations, and the evaluation of the feasibility of PFASs substitute applications.
... According to earlier research, the PFOA 96-h LC50 values in zebrafish were around 500 mg/L [24][25][26]. In another study, the PFOA 96-h LC50 for larval rainbow trout was 730 mg/L [27]. ...
PFOA, a newly emerging persistent organic pollutant, is widely present in various environmental media. Previous reports have proved that PFOA exposure can accumulate in the ovary and lead to reproductive toxicity in pregnant mice. However, the potential mechanism of PFOA exposure on fertility remains unclear. In this study, we explore how PFOA compromises fertility in the zebrafish. The data show that PFOA (100 mg/L for 15 days) exposure significantly impaired fertilization and hatching capability. Based on tissue sections, we found that PFOA exposure led to ovarian damage and a decrease in the percentage of mature oocytes. Moreover, through in vitro incubation, we determined that PFOA inhibits oocyte development. We also sequenced the transcriptome of the ovary of female zebrafish and a total of 284 overlapping DEGs were obtained. Functional enrichment analysis showed that 284 overlapping DEGs function mainly in complement and coagulation cascades signaling pathways. In addition, we identified genes that may be associated with immunity, such as LOC108191474 and ZGC:173837. We found that exposure to PFOA can cause an inflammatory response that can lead to ovarian damage and delayed oocyte development.
... Supplementary Materials: The following supporting information can be downloaded at: https: //www.mdpi.com/article/10.3390/foods12071423/s1, Table S1: Acute toxicity data [60][61][62][63][64][65][66][67][68][69][70][71][72][73][74]; Table S2: The gradient elution program; Table S3: MRM experimental conditions used for LC-MS/MS identification of PFOA and known metabolites in fish organs and tissue; Table S4: Recovery yields (R), matrix effect (EM), and limits of quantification (LOQ) obtained for PFOA and its metabolites in fish tissue; Table S5: Initial characteristics of the specimens selected for the bioconcentration test (0 days); Table S6: Final characteristics of the specimens after the exposure period (92 days); Table S7: PFOA concentration values determined in tissues and organs in Experiment 1 (10 µg/L PFOA); Table S8: PFOA concentration values determined in tissues and organs in Experiment 2 (100 µg/L PFOA); Table S9: Concentration values (ng/g d.w.) of PFOA biotransformation products determined in fish organs; Table S10: Spearman correlation and p-values determined between PFOA and metabolites in the gills; Table S11: Spearman correlation and p-values determined between PFOA and metabolites in the kidney; Table S12: Spearman correlation and p-values determined between PFOA and metabolites in the brain; Table S13: Spearman correlation and p-values determined between PFOA and metabolites in the gonads; Table S14: Spearman correlation and p-values determined between PFOA and metabolites in the liver; Table S15: Spearman correlation and p-values determined between PFOA and metabolites in the gallbladder; Table S16: Spearman correlation and p-values determined between PFOA and metabolites in the intestine; Table S17: Spearman correlation and p-values determined between PFOA and metabolites in the muscle; Table S18: Spearman correlation and p-values determined between PFOA and metabolites in the skin; Table S19: Spearman correlation and p-values determined between PFOA and metabolites in the scales; Figure S1: MRM chromatogram obtained for PFOA and selected metabolites (10 µg/L); Figure S2. The total concentration of PFOA (a) and the percentage distribution in organs (b) in Experiment 1; Figure S3: The total concentration of PFOA (a) and the percentage distribution in organs (b) in Experiment 2; Figure S4: PFOA variation and biotransformation product evolution in fish organs in Experiment 1; Figure S5: PFOA variation and biotransformation product evolution in fish organs in Experiment 2. ...
The perfluoroalkyl substances (PFAS) represent a persistent class of synthetic chemicals that spread in the environment as a result of industrialization. Due to their bioaccumulative and endocrine disruption implications, these chemicals can affect food quality and human health, respectively. In the present study, the bioconcentration and biotransformation of perfluorooctanoic acid (PFOA) in common carp (Cyprinus carpio) were evaluated in a biphasic system (exposure and depuration). Carp were continuously exposed, under laboratory conditions, to 10 (Experiment 1) and 100 (Experiment 2) µg/L PFOA for 14 weeks, followed by a wash out period of 3 weeks. Fish organs and tissues were collected at 8, 12, 14 weeks of exposure and at week 17, after the depuration period. The results obtained from the LC-MS/MS analysis showed the presence of PFOA in all studied organs. The highest values of PFOA were identified in the gallbladder (up to 2572 ng/g d.w.) in Experiment 1 and in the gallbladder (up to 18,640 ng/g d.w.) and kidneys (up to 13,581 ng/g d.w.) in Experiment 2. The average BCF varied between 13.4 and 158 L/Kg in Experiment 1 and between 5.97 and 80.3 L/Kg in Experiment 2. Four biotransformation products were identified and quantified in all organs, namely: PFBA, PFPeA, PFHxA, and PFHpA. PFBA was proven to be the dominant biotransformation product, with the highest values being determined after 8 weeks of exposure in the kidney, gallbladder, brain, liver, and gonads in both experiments. Because freshwater fish are an important food resource for the human diet, the present study showed the fishes’ capacity to accumulate perfluoroalkyl substances and their metabolites. The study revealed the necessity of monitoring and risk studies of new and modern synthetic chemicals in aquatic resources.
... 48-78 (A) or 47-61 (C) organisms from all samples were measured across 3 independent experiments . was also reported in other freshwater organisms, e.g., PFAS shared comparable EC 50 toxicities between T. pyriformis (PFOA: 1724 μM / 714 mg/L and PFOS: 103 μM / 52 mg/L), green neon shrimps (Neocaridina denticulate); PFOA: 2400 μM / 1000 mg/L and PFOS: 400 μM / 200 mg/L) water fleas (Daphnia magna; PFOA: 720 μM / 298 mg/L and PFOS: 386 μM / 193 mg/L; Moina macrocopa; PFOA: 481 μM / 199.51 mg/L and PFOS: 36 μM / 18 mg/L) and zebrafish (Danio rerio; PFOA: 2427 μM / 1005 mg/L and PFOS: 214 μM / 107 mg/L) (Ji et al., 2008;Li, 2009;Ye et al., 2009). ...
Per-/Poly- fluoroalkyl substances represent emerging persistent organic pollutants. Their toxic effects can be broad, yet little attention has been given to organisms at the microscale. To address this knowledge shortfall, the unicellular eukaryote Tetrahymena pyriformis was exposed to increasing concentrations (0 - 5000 μM) of PFOA/PFOS and monitored for cellular motility, division and function (i.e., phagocytosis), reactive oxygen species generation and total protein levels. Both PFOA/PFOS exposure had negative impacts on T. pyriformis, including reduced motility, delayed cell division and oxidative imbalance, with each chemical having distinct toxicological profiles. T. pyriformis represents a promising candidate for assessing the biological effects these emerging anthropogenically-derived contaminants in a freshwater setting.
... PFOA was able to have multiple effects on the endocrine system by interfering with hormone receptors ER and TR, altering steroid hormone synthesis and major steroidogenic gene expression, and disrupting endocrine-related gene levels in zebrafish at 0.1-0.5 mg/L from 4 h postfertilization (hpf) to 120 hpf (Du et al., 2013). In addition, the 96 h LC 50 values of PFOS and PFOA were 71 mg/L and 499 mg/L, respectively (Lu et al., 2009). In our study, the 96 h LC 50 of PFHxA was calculated to be 79.72 mg/L in our experiment by using the 24-hole plate method (EPA et al., 1996). ...
Perfluorohexanoic acid (PFHxA) has been recognized as an alternative to the wide usage of perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) in the fluoropolymer industry for years. PFHxA has been frequently detected in the environment due to its wide application. However, the ecological safety of PFHxA, especially its toxicological effects on aquatic organisms, remains obscure. In the present study, PFHxA at different concentrations (0, 0.48, 2.4, and 12 mg/L) was added to the culture medium for zebrafish embryo/larval exposure at 96 h postfertilization (hpf). Zebrafish larvae showed a slow body growth trend and changes in thyroid hormone levels (THs) upon PFHxA exposure, indicating the interference effect of PFHxA on fish larval development. Moreover, the transcription levels of genes related to the hypothalamic-pituitary-thyroid (HPT) axis were also analyzed. The gene expression level of thyroid hormone receptor β (trβ) was upregulated in a dose-dependent manner. Exposure to 0.48 mg/L PFHxA increased the expression levels of the thyrotrophic-releasing hormone (trh) and thyroid hormone receptor α (trα). Significant increases in corticotrophin-releasing hormone (crh) and transthyretin (ttr) gene expression were also observed when the zebrafish larvae were treated with 12 mg/L PFHxA, except iodothyronine deiodinases (dio1), which decreased obviously at that point. There were significant declines in the transcription of both thyroid-stimulating hormone β (tshβ) and uridinediphosphate-glucuronosyltransferase (ugt1ab) upon exposure to 2.4 mg/L PFHxA. In addition, PFHxA induced a dose-related inhibitory effect on the transcription of sodium/iodide symporter (nis). Finally, the thyroid status will be destroyed after exposure to PFHxA, thus leading to growth impairment in zebrafish larvae.
... LC 50 (96 h) values of PFOA in zebrafish were approximately 500 mg/ L (Ye et al., 2009;Valsecchi et al., 2017). 1/20 of LC 50 was chosen to obtain more stable and accurate content of PFOA in different tissues in the early stage of exposure while reducing the influence on the survival growth of zebrafish (Narra et al., 2011;Appenzeller et al., 2016;Levitskaia et al., 1985). ...
Perfluorooctanoic acid (PFOA) as an emerging environmental contaminant, has become ubiquitous in the environment. It is of significance to study bioconcentration and tissue distribution of aquatic organisms for predicting the persistence of PFOA and its adverse effects on the environment and human body. However, the distribution of PFOA in different tissues is a complex physiological process affected by many factors. It is difficult to be accurately described by a simple kinetic model. In present study, a new strategy was introduced to research the PFOA distribution in tissues and estimate the exposure stages. Zebrafish were continuously exposed to 25 mg/L PFOA for 30 days to simulate environmental process. Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) method was used to monitor the spatio-temporal distribution of PFOA in zebrafish tissues. By analyzing the law of change obtained from the high spatial resolution MSI data, two different enrichment trends in ten tissues were summarized by performing curve fitting. Analyzing the ratio of two types of curves, a new “exposure curve” was defined to evaluate the exposure stages. With this model, three levels (mild, moderate, and deep pollution stage) of PFOA pollution in zebrafish can be simply evaluated.
... The initial PFOS concentrations in water were held approximately constant with respect to the particles size, with a concentration to particle size ratio of 0.6 to 0.8. The concentration of PFOS obtained on PE MPs particles at low sorption, e.g., ng/g range, corresponded to environmental concentration that were found in sewage sludge (Loganathan et al. 2007;Becker et al. 2008;Guo et al. 2009), whereas the PFOS concentrations at high sorption concentrations tends to represent a concentration inducing acute toxicity on zebrafish embryos exposed to the pure compound (Ye et al. 2009). The effect of the particle size on the sorption capacity of PE MPs is poorly understood, but it has been shown that a decrease in particles size, at micron and submicron size, increased the sorption rates of contaminants (Wang et al. 2019). ...
The sorption processes of persistent organic pollutants on microplastics particles are poorly understood. Therefore, the present study investigated the sorption processes of perfluorooctanesulfonate (PFOS) on polyethylene (PE) microplastic particles (MPs) which are representing a prominent environmental pollutant and one of the most abundant microplastic polymers in the aquatic environment, respectively. The focus was set on the investigation of the impact of the particle size on PFOS sorption using four different PE MPs size ranges. The sorption kinetics for 6 months was studied with one selected size range of PE MPs. Besides, the desorption of PFOS from PE MPs under simulated digestive conditions was carried out by using artificial gut fluid mimicking the intestinal juice of fish. The investigation of the size effects of particles over 6 months demonstrated a linear increase of PFOS concentration sorbed onto PE with a decrease of the particle size. Thus, our findings implicate efficient sorption of PFOS onto PE MPs of different sizes. The results showed that PFOS desorbed from the PE MPs into the artificial gut fluid with a rate of 70 to 80%. Besides, a longer exposure of PE MPs to PFOS leads to a higher concentration adsorbed by PE MPs, which may favor the ingestion of higher concentration of PFOS, and thus represents a higher risk to transfer relevant concentrations of PFOS during digestion.
Per- and polyfluoroalkyl substances (PFAS) are commonly used as surfactants and coatings for industrial processes and consumer products. These compounds have been increasingly detected in drinking water and human tissue, and concern over their potential effects on health and development is growing. However, relatively little data are available for their potential impacts on neurodevelopment and the degree to which different compounds within this class may differ from one another in their neurotoxicity. The present study examined the neurobehavioral toxicology of two representative compounds in a zebrafish model. Zebrafish embryos were exposed to 0.1-100uM perfluorooctanoic acid (PFOA) or 0.01-1.0uM perfluorooctanesulfonic acid (PFOS) from 5 to 122 h post-fertilization. These concentrations were below threshold for producing increased lethality or overt dysmorphologies, and PFOA was tolerated at a concentration 100× higher than PFOS. Fish were maintained to adulthood, with behavioral assessments at 6 days, 3 months (adolescence) and 8 months of age (adulthood). Both PFOA and PFOS caused behavioral changes in zebrafish, but PFOS and PFOS produced strikingly different phenotypes. PFOA was associated with increased larval motility in the dark (100uM), and enhanced diving responses in adolescence (100uM) but not adulthood. PFOS was associated with a reversed light-dark response in the larval motility test (0.1-1uM), whereby the fish were more active in the light than the dark. PFOS also caused time-dependent changes in locomotor activity in the novel tank test during adolescence (0.1-1.0uM) and an overall pattern of hypoactivity in adulthood at the lowest concentration (0.01uM). Additionally, the lowest concentration of PFOS (0.01uM) reduced acoustic startle magnitude in adolescence, but not adulthood. These data suggest that PFOS and PFOA both produce neurobehavioral toxicity, but these effects are quite distinct from one another.
Perfluoroalkyl substances (PFAS) are highly persistent organic pollutants that have been detected in a wide array of environmental matrices and, in turn, diverse biota including humans and wildlife wherein they have been associated with a multitude of toxic, and otherwise adverse effects, including ecosystem impacts. In the present study, we developed a toxicity assay for embryonic stages of mahi-mahi (Coryphaena hippurus), as an environmentally relevant pelagic fish species, and applied this assay to the evaluation of the toxicity of “legacy” and “next-generation” PFAS including, respectively, perfluorooctanoic acid (PFOA) and several perfluoroethercarboxylic acids (PFECA). Acute embryotoxicity, in the form of lethality, was measured for all five PFAS toward mahi-mahi embryos with median lethal concentrations (LC50) in the micromolar range. Consistent with studies in other similar model systems, and specifically the zebrafish, embryotoxicity in mahi-mahi generally (1) correlated with fluoroalkyl/fluoroether chain length and hydrophobicity, i.e., log P, of PFAS, and thus, aligned with a role of uptake in the relative toxicity; and (2) increased with continuous exposure, suggesting a possible role of development stage specifically including a contribution of hatching (and loss of protective chorion) and/or differentiation of target systems (e.g., liver). Compared to prior studies in the zebrafish embryo model, mahi-mahi was significantly more sensitive to PFAS which may be related to differences in either exposure conditions (e.g., salinity) and uptake, or possibly differential susceptibility of relevant targets, for the two species. Moreover, when considered in the context of the previously reported concentration of PFAS within upper sea surface layers, and co-localization of buoyant eggs (i.e., embryos) and other early development stages (i.e., larvae, juveniles) of pelagic fish species to the sea surface, the observed toxicity potentially aligns with environmentally relevant concentrations in these marine systems. Thus, impacts on ecosystems including, in particular, population recruitment are a possibility. The present study is the first to demonstrate embryotoxicity of PFAS in a pelagic marine fish species, and suggests that mahi-mahi represents a potentially informative, and moreover, environmentally relevant, ecotoxicological model for PFAS in marine systems.
Acute toxicity testing in fish is a standard method used in estimating the influences of chemicals on aquatic vertebrates. The ecotoxicological data obtained from acute toxicity tests in fish are, however, not sufficiently reliable to justify the continued use of this test.
Fertilised eggs of zebrafish ( Brachydanio rerio) were used to test the acute toxicity of chemicals. They were chosen because the development of B. rerio has been studied extensively and information already exists concerning the normal development of this species.
The following parameters of the development of B. rerio were observed: coagulation of the egg, gastrulation, number of somites, movement, development of organs, pigmentation, heartbeat and circulation. Some of these toxicological endpoints indicate lethality. In addition, various interactions between the test chemical and the embryos can be measured by investigating these parameters.
Six chemicals (2,4-dinitrophenol, 4,6-dinitro-o-cresol, malathion, carbaryl, phenol and 4-nitrophenol) were selected for testing in the embryos of zebrafish within the first 48 hours of their development. The toxicities of the test chemicals to zebrafish embryos were compared with their acute toxicities to adult fish. Further investigations with more chemicals are in progress.
The effective concentrations (EC50) and the lowest effect concentrations (LOEC) investigated in this preliminary study were comparable to the LC50 values for adult fish. The use of this test system as an alternative method to testing toxicity in adult fish is discussed.
The acute biological activity of a homologous series of perfluorinated carboxylic acids - perfluorohexanoic acid (PFHxA), perfluoroheptanoic acid (PFHpA), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA) and perfluorodecanoic acid (PFDA) - was studied. To analyze the potential risk of the perfluorinated acids to humans and the environment, different in vitro toxicity test systems were employed. The cytotoxicity of the chemicals towards two different types of mammalian cell lines and one marine bacteria was investigated. The viability of cells from the promyelocytic leukemia rat cell line (IPC-81) and the rat glioma cell line (C6) was assayed calorimetrically with WST-1 reagent. The evaluation was combined with the Vibrio fischeri acute bioluminescence inhibition assay. The biological activity of the compounds was also determined at the molecular level with acetylcholinesterase and glutathione reductase inhibition assays. This is the first report of the effects of perfluorinated acids on the activity of purified enzymes. The results show these compounds have a very low acute biological activity. The observed effective concentrations lie in the millimole range, which is well above probable intracellular concentrations. A relationship was found between the toxicity of the perfluorinated carboxylic acids and the perfluorocarbon chain length: in every test system applied, the longer the perfluorocarbon chain, the more toxic was the acid. The lowest effective concentrations were thus recorded for perfluorononanoic and perfluorodecanoic acids.
Acute toxicity of BDR-29, a combined preparation of Cassia Semen, Prunellae Spica, Tribuli Fructus, and Uncariae Rhamulus et Uncus was examined using male and female Sprague-Dawley rats. Rats were treated with the BDR-29 intiagastrically at 0 mg/kg, 5 mg/kg, 50 mg/kg, 500 mg/kg or 2,000 mg/kg and observed for two weeks. At the doses used no mortality or abnormal clinical signs in animals were shown during at the observation period. Also there was no difference in net body weight gain, gross pathological findings, and urine analysis at the terminal sacrifice among the groups rats treated wih different doses of the test substance. The results suggested that acute oral toxicity of BDR-29 in rats is very low at the conditions employed in this study.
We describe a series of stages for development of the embryo of the zebrafish, Danio (Brachydanio) rerio. We define seven broad periods of embryogenesis—the zygote, cleavage, blastula, gastrula, segmentation, pharyngula, and hatching periods. These divisions highlight the changing spectrum of major developmental processes that occur during the first 3 days after fertilization, and we review some of what is known about morphogenesis and other significant events that occur during each of the periods. Stages subdivide the periods. Stages are named, not numbered as in most other series, providing for flexibility and continued evolution of the staging series as we learn more about development in this species. The stages, and their names, are based on morphological features, generally readily identified by examination of the live embryo with the dissecting stereomicroscope. The descriptions also fully utilize the optical transparancy of the live embryo, which provides for visibility of even very deep structures when the embryo is examined with the compound microscope and Nomarski interference contrast illumination. Photomicrographs and composite camera lucida line drawings characterize the stages pictorially. Other figures chart the development of distinctive characters used as staging aid signposts. ©1995 Wiley-Liss, Inc.
In the present study we evaluated the toxicological effects of a scarcely documented environmental pollutant, perfluorooctane sulfonic acid (PFOS), on selected biochemical endpoints in the common carp, Cyprinus carpio. Juvenile organisms were exposed to PFOS through a single intraperitoneal injection (liver concentrations ranging from 16 to 864 ng/g after 5 days of exposure) and after 1 and 5 days effects were assessed in liver and serum of the exposed organisms. The investigation of the hepatotoxicity of PFOS included the determination of the peroxisome proliferating potential (peroxisomal palmitoyl CoA oxidase and catalase activity) and the compounds influence on the average DNA basepair length (ABPL) by agarose gel electrophoresis. Total antioxidant activity (TAA), cholesterol and triglyceride levels were monitored in the serum. After 1 day of exposure the ABPL was significantly increased in the 270 and 864 ng/g treatment groups. After 5 days of exposure significant increases relative to the control were observed for the 16, 270 and 864 ng/g treatment groups. Enzyme leakage from the liver was investigated by measurement of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in the serum. At 561, 670 and 864 ng/g PFOS a significant increase in serum ALT activity became apparent after 5 days of exposure with values ranging from 159 to 407% relative to the control. For serum AST activity a significant increase for the 864 ng/g treatment group was observed with a value of 112% relative to the control. Determination of the polymorphonuclear leukocyte migration into liver tissue as assessed through myeloperoxidase (MPO) activity in liver, was used as an indicator for inflammation. It appeared that inflammation was not involved in the observed membranous enzyme leakage for the 561, 670 and 864 ng/g PFOS treatment groups. The results of this study suggest that PFOS induces inflammation-independent enzyme leakage through liver cell membranes that might be related to cell necrosis. Furthermore, results show that PFOS does not significantly affects serum antioxidant levels nor does it clearly induce peroxisome proliferation in carp. This study also points out that PFOS might interfere with homeostasis of the DNA metabolism. The results of these biochemical analyses were used to perform an initial hazard assessment study indicating that PFOS levels observed in tissues of wildlife populations could induce a clear rise in serum transaminase levels indicative for disruption of hepatocyte membrane integrity.
Perfluorooctanoic acid (PFOA) is an octanoic acid derivative to which all aliphatic hydrocarbons are substituted by fluorine. PFOA and its salts are commercially used in various industrial processes. The chemical is persistent in the environment and does not undergo biotransformation. It was reported that PFOA is found not only in the serum of occupationally exposed workers but also general populations. Recent studies have suggested that the biological half-life of PFOA in humans is 4.37 years based on study of occupationally exposed workers. It is increasingly suspect that PFOA accumulates and affects human health, although the toxicokinetics of PFOA in humans remain unclear. In experimental animals, PFOA seems low in toxicity. PFOA is well-absorbed following oral and inhalation exposure, and to a lesser extent following dermal exposure. Once absorbed in the body, it distributes predominantly to the liver and plasma, and to a lesser extent the kidney and lungs. PFOA is excreted in both urine and feces. Biological half-life of PFOA is quite different between species and sexes and the difference is due mainly to the difference in renal clearance. In rats, renal clearance of PFOA is regulated by sex hormones, especially testosterone. PFOA is excreted into urine by active tubular secretion, and certain organic anion transporters are though to be responsible for the secretion. Fecal excretion is also important in the elimination of PFOA. There is evidence that PFOA undergoes enterohepatic circulation resulting in reduced amounts of fecal excretion. Elucidation of the mechanisms of transport in biological systems leads to elimination and detoxification of this chemical in the human body.
This study investigates reproductive impairment and biochemical changes in fathead minnow (Pimephales promelas) exposed for 39 d to varying concentrations of perfluorooctanoic acid (PFOA) under microcosm conditions. While the concentrations tested in this study were much higher than those normally found in the environment, no mortality was associated with PFOA exposure. Only modest changes were observed in condition factor and in relative liver and gonad size. Significant declines in circulating plasma steroids were observed, but these were accompanied by only limited increases in time to first oviposition and decreases in overall egg production. Peroxisome proliferation, as quantified by fatty acyl-CoA oxidase (FAO) activity, was elevated with low PFOA concentrations but attenuated with exposure to higher PFOA doses. Little evidence was seen of differential induction of peroxisome-associated enzyme activity with sex. Oxidative stress, as quantified by the 2-thiobarbituric acid reactive substances (TBARS) assay, was only modestly influenced by PFOA exposure and is not a significant consequence of FAO activity in fathead minnow. Perfluorooctanoic acid appears to be relatively nontoxic at environmentally relevant concentrations but may impact biochemical and reproductive endpoints under conditions associated with environmental spills.
Rainbow trout (Onchorhynchus mykiss) liver microsomes were incubated with N-ethyl perfluorooctanesulfonamide [N-EtPFOSA, C8F17SO2NH(C2H5)], to examine the possibility of in vitro biotransformation to perfluorooctane sulfonate (PFOS, C8F17SO3-) and perfluorooctanoate (PFOA, C7F15COO-). Incubations were performed by exposing trout liver microsomes to N-EtPFOSA at 8 degrees C in the dark. Reaction mixtures were analyzed after incubation periods of 0, 2, 4, 8, 16, and 30 h for N-EtPFOSA, PFOS, PFOA, and perfluorooctanesulfonamide (PFOSA, C8F17SO2NH2), a suspected intermediate. Amounts of PFOS and PFOSA were found to increase with incubation time, but only background levels of PFOA were detected. Three possible reaction pathways are proposed for the conversion of N-EtPFOSA to PFOS: (i) direct conversion of N-EtPFOSA to PFOS by deethylamination accompanied by conversion of the sulfone group to sulfonate, (ii) deethylation of N-EtPFOSA to PFOSA, followed by deamination to form PFOS, and (iii) direct hydrolysis of N-EtPFOSA. These findings represent the first report indicating a possible biotransformation of a perfluorosulfonamide to PFOS in fish and may help to explain the detection of PFOS, which is relatively involatile, and thus not likely to undergo atmospheric transport, in biota from remote regions.
Perfluorooctanoic acid (PFOA) is an organic fluorochemical, and its elimination in rats is markedly sex-dependent. Liver and kidney are two primary tissues of distribution of PFOA in rats. In this study, the subcellular distribution of PFOA in male and female rat liver and kidney was examined. The results demonstrated that PFOA content in the liver cytosol of the female rat was significantly higher (49 +/- 6% of total radioactive residues, TRR) than in the male liver (26 +/- 5% TRR), whereas PFOA distribution in the heavier subcellular fractions, especially the nuclei and cell debris fraction, was marginally higher in male rat liver. In rat kidney, more than 70% of PFOA was distributed in the cytosolic fraction, with no significant difference between sexes. The degree of protein binding of PFOA in rat liver and kidney cytosol was analyzed by two different chromatographic methods. The percentage of protein-bound PFOA in the liver cytosol was found to be approximately 55% in both male and female rats. In contrast, significantly more PFOA was bound to cytosolic proteins in the kidney of male rats (42 +/- 6% TRR) than in females (17 +/- 5% TRR). Ligand blotting analysis revealed that multiple proteins from the liver cytosol, nuclei, and mitochondria fractions were capable of specific binding to PFOA.
Perfluorooctanoic acid (PFOA) is an anthropogenic contaminant detected in various environmental and biological matrices. This compound is a fluorinated surfactant, belonging to a class of molecules known for persistence and their global distribution, but for which little ecotoxicological data are currently available, especially under field conditions. The environmental fate and toxicity of PFOA to the aquatic macrophytes Myriophyllum sibiricum and M. spicatum were investigated using 12,000 L outdoor microcosms. Replicate microcosms (n = 3) were treated with 0.3, 1, 30, and 100 mg/L PFOA as the sodium salt, plus controls, and assessed at regular intervals over 35 days. PFOA showed no significant dissipation from the water column, except at the greatest concentration, where partitioning from the water column into other compartments is suspected. The two species of Myriophyllum were similar in their sensitivity to PFOA under these simulated field conditions. Toxicity after 14 to 35 days of exposure in the evaluated endpoints for M. spicatum was > or = 5.7 mg/L PFOA for EC10s and > or = 31.8 mg/L PFOA for EC50s and in M. sibiricum was > or = 8.4 mg/L PFOA for EC10s and > or = 35.8 mg/L PFOA for EC50s. The no observed effects concentrations (NOECs) for Myriophyllum spp. were consistently > or = 23.9 mg/L PFOA. A risk assessment for these plant species estimated a negligible probability of toxicity being observed from PFOA exposure at current environmental concentrations.