Toxicities of some heavy metals to the tadpoles of frog, Microhyla ornata (Dumeril & Bibron)

Static bioassays were conducted to determine the relative acute toxicities of five heavy metals (mercury, cadmium, copper, manganese and zinc) to 1-week-old and 4-week-old tadpoles of the frog, Microhyla ornata. Toxic effects were calculated on the basis of LC50 for 24 h, 48 h, 72 h and 96 h exposures at 25.5-26.0 degrees C. Mercury was the most toxic and zinc was the least toxic of the heavy metals tested. The sensitivity of the tadpoles to the heavy metals increased with increased age.     

Sublethal effects of chronic exposure to an organochlorine compound on northern leopard frog (Rana pipiens) tadpoles

Global contamination with organochlorine compounds (OCs) has posed developmental and reproductive problems in wildlife worldwide. However, little is known about the impact of OCs or other pollutants on amphibians, despite mounting concerns about amphibian population declines and developmental deformities in the wild. Wildlife populations may be affected critically by sublethal impacts of anthropogenic disturbances, yet little research has focused on such effects in amphibians. In the current study, northern leopard frog (Rana pipiens) tadpoles were chronically exposed to a polychlorinated biphenyl (PCB) congener, 77-TCB, and effects on behavior, morphology, competitive performance, and corticosterone content were determined. R. pipiens activity levels and feeding rates were decreased by 77-TCB exposure, but morphology of mouthparts and body proportions were unaffected. 77-TCB enhanced growth and altered competitive interactions between R. pipiens and wood frog (Rana sylvatica) tadpoles. R. pipiens tadpoles exposed to 77-TCB showed decreased whole-body corticosterone content compared to controls both before and after injection with adrenocorticotropic hormone (ACTH). All of the factors examined in the current study play critical roles in tadpole development, growth, survivorship, and eventual reproductive success, suggesting negative population-level consequences for amphibians in PCB-contaminated habitats.     

GC法同时测定林蛙卵中5种脂肪酸含量

目的:建立林蛙卵中肉豆蔻酸、棕榈酸、硬脂酸、油酸和亚油酸的含量测定方法。方法:采用毛细管气相色谱法,甲酯化后同时测定林蛙卵中肉豆蔻酸、棕榈酸、硬脂酸、油酸和亚油酸的含量。采用PEG-20M毛细管柱(30 m×0.25 mm,0.25μm);氢火焰离子化检测器(FID);程序升温:起始温度200℃,保持2 m in,以4℃.m in-1升至230℃,维持8 m in;进样口温度:260℃;检测器温度:280℃;流速:0.3 mL.m in-1;分流比为20∶1;进样体积为1μL。结果:肉豆蔻酸甲酯、棕榈酸甲酯、硬脂酸甲酯、油酸甲酯和亚油酸甲酯浓度分别在0.015~0.152 mg.mL-1(r=0.9994),0.106~1.056 mg.mL-1(r=0.9993),0.020~0.203 mg.mL-1(r=0.9991),0.133~1.332 mg.mL-1(r=0.9995),0.055~0.545 mg.mL-1(r=0.9992)范围内有良好的线性关系(n=6);方法回收率(n=9)分别为97.4%,97.6%,97.9%,97.4%,96.0%。结论:本法简单、准确,重复性好,可作为测定林蛙卵中...     

Toxicity of endosulfan to tadpoles of Fejervarya spp. (Anura: Dicroglossidae): mortality and morphological deformities

The acute toxicity of endosulfan to the tadpoles of three coexisting species of the anuran genus Fejervarya revealed 96 h LC₅₀ values of 46.715, 6.596, and 3.015 μg l^?1 for Fejervarya sp.1, F. teraiensis and Fejervarya sp.2, respectively. Toxicity of endosulfan was also tested at the sublethal concentrations of 5 and 0.5, and 0.3 and 0.03 μg l^?1 (c 10 and 1 % of their respective 96 h LC₅₀ values) in Fejervarya sp.1 and Fejervarya sp.2, and 0.35 and 0.18 μg l^?1 (c 5 and 2.5 % of 96 h LC₅₀) in F. teraiensis. Endosulfan was observed to cause mortality at concentrations as low as c 1, 2.5 and 10 % of their respective 96 h LC₅₀ values in Fejervarya sp.2, F. teraiensis, and Fejervarya sp.1. Such vulnerabilities are likely to have implications for the survival of natural populations of these co-existing species as well as other anurans present in the study area where pesticide use is relatively high in the tea plantations. Morphological deformities caused by endosulfan comprised failure to develop one or both forelimb in Fejervarya sp.1 and F. teraiensis, stunted hindlimb growth in Fejervarya sp.1, and axial malformation in Fejervarya sp.1 and Fejervarya sp.2. Fore- and hind-limb deformities were likely to have occurred due to the impairment of thyroid metabolism by endosulfan. These effects illustrate the threat that continued endosulfan use poses to natural populations of anuran amphibians.     

Assessment of antioxidant system,cholinesterase activity and histopathology in Lithobates catesbeianus tadpoles exposed to water from an urban stream

Ecotoxicology - Anthropogenic activities promote changes in community structure and decrease the species abundance of amphibians. The aim of this study was to assess potential alterations in the...     

Effects of exposure to naphthenic acids in tree swallows (Tachycineta bicolor) on the Athabasca oil sands, Alberta, Canada

Naphthenic acids (NAs) are a group of carboxylic acids that are of particular concern to the steadily growing oil sands mining industry of Alberta, Canada, because they become highly concentrated in the water used for oil sands extraction and are toxic to aquatic biota and mammals. Upon mine closure, vast amounts of process-affected water will need to be reclaimed and proven safe for wildlife colonizing reclaimed areas. The effects of exposure to NAs have not been investigated in avian species. To address this void, tree swallow (Tachycineta bicolor) nestlings were dosed with NAs while being reared normally by their free-ranging parents on a site in the vicinity of the oil sands. Nestlings received 1.5 mg NAs/day (approximately 0.075 g/kg body mass) from d 7 to d 13 of age, which represented a 10-fold "worst exposure" scenario. Nestling growth, hematocrit, blood biochemistry, organ weights, and ethoxyresorufin O-deethylase (EROD) activity were unaffected by NAs. The only change detected on histopathological evaluation of major organs was an increase in extramedullary erythropoiesis in the liver. These findings indicate that nestling tree swallows can successfully tolerate short-term exposures to environmentally realistic concentrations of NAs. However, this study did not investigate the chronic or reproductive toxicity of NAs. More research needs to be conducted to complete this initial assessment, to determine environmental risks on reclaimed areas where birds will be breeding and where their exposure to NAs could extend for several weeks.     

Oxidative damage of naphthenic acids on the Eisenia fetida earthworm

Naphthenic acids (NAs) have been gaining recognition in recent years as potentially harmful environmental contaminants. Few studies have focused on the potential ecotoxicity of NAs to terrestrial environment. In this study, the responses of antioxidant system and lipid peroxidation and DNA damage were investigated after exposing Eisenia fetida to soil contaminated with NAs. The results indicated that NAs induced a significant increase (p < 0.05) in superoxide dismutase and catalase enzyme activities. The glutathione peroxidase enzyme activities were significantly inhibited (p < 0.05) in the medium and high dose treatments. An increase in malondialidehyde indicated that NAs could cause cellular lipid peroxidation in the tested earthworms. The percentage of DNA in the tail of comet assay of coelomocytes as an indication of DNA damage increased after treatment with different doses of NAs, and a dose‐dependent DNA damage of coelomocytes was found. In conclusion, oxidative stress caused by NAs exposure induces physiological responses and genotoxicity on earthworms. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1337–1343, 2016.     

Toxicity of nitromusks in early lifestages of South African clawed frog (Xenopus laevis) and zebrafish (Danio rerio)

Musk xylene (MX), musk ketone (MK) and musk moskene (MM) are synthetic nitro-containing fragrances. Due to their inherent lipophilicity and environmental persistence, they are frequently detected in environmental samples and especially in aquatic ecosystems. Despite this, the current environmental toxicity database of nitromusks is limited. Although nitromusks have been shown to accumulate in aquatic organisms, little is known about their potential developmental effects in the respective aquatic species. To investigate the developmental toxicity of these compounds to amphibians and fish, early lifestages of xenopus (Xenopus laevis) and zebrafish (Danio rerio) were exposed to three nitromusks for 96 h to examine the developmental effects of these compounds in the two species. Nitromusk body concentration measurements were carried out in parallel for correlation with potential developmental effects. No increased mortality, malformation or growth inhibition was observed in either species following 96-h exposure to 400 microg/l MX, MK and MM. However, an approximately 20% reduced viability was observed in xenopus larvae when exposed to 400 microg/l MX, MK and MM for 11 days. Xenopus and zebrafish exposed to 10, 153, 871 and 1637 microg/l 14C-MX for 96 h resulted in whole-body concentrations of 0.7 +/- 0.1, 11.1 +/- 1.1, 38.7 +/- 1.9 and 76.3 +/- 18.3 microg/g, and 4.3 +/- 0.6, 73.3 +/- 11.8, 440.0 +/- 72.7 and 664.0 +/- 47.7 microg/g wet body weight, respectively. Exposure of xenopus larvae to 400 microg/l MX, MK and MM for 11 days, resulted in whole body concentrations (extrapolated from gas chromatographic determinations) of 4700 +/- 5000, 1300 + 300 and 4600 + 4800 microg/g wet weight for MX, MK and MM, respectively. The latter toxicity results, in conjunction with the fact that the concentrations used for the above experiments were between 400- and 10000-fold higher than those detected in the environment, suggest that environmental concentrations of nitromusks are not hazardous for early lifestages of fish and amphibians.     

Combined effects of malathion and nitrate on early growth,abnormalities, and mortality of wood frog (<Emphasis Type="Italic">Rana sylvatica</Emphasis>) tadpoles

Use of pesticides and other agro-chemicals adversely influence amphibians either directly by killing them or by inducing sublethal, chronic effects. Many studies have investigated the effect of mixtures of pesticides or fertilizers. We studied the combined effects of nitrate and malathion ([(dimethoxy phosphino thioyl] butanediotae) on the early growth, expression of abnormalities, and mortality of Wood Frog (Rana sylvatica) tadpoles in a laboratory experiment. Tadpoles were treated with factorial combinations of 0, 8, and 16 mg NO₃–N l⁻¹ and 0, 250, 500, and 1,000 μg malathion l⁻¹ for a period of 14 days. Feeding behaviour, total length, mean tadpole mass, frequencies of abnormalities, and survivorship in each treatment were recorded. Malathion showed a significant negative influence on all parameters and strongly influenced the frequencies of morphological anomalies. In contrast, nitrate alone did not produce any significant effects on behavior, total length, tadpole mass, or the frequency of abnormalities during the experiment. Malathion and nitrate had an interactive effect on tadpole length and mass, but did not affect any other parameters. Our results suggest that exposure to malathion, even at relatively low concentrations can have serious negative consequences for Wood Frog tadpoles. In addition, our results also indicate that there was little synergistic interaction between malathion and nitrate exposure under laboratory conditions.     

Acute and subchronic mammalian toxicity of naphthenic acids from oil sands tailings.

Naphthenic acids are the most significant environmental contaminants resulting from petroleum extraction from oil sands deposits. In this study, a mixture of naphthenic acids isolated from Athabasca oil sands (AOS) tailings pond water was used in acute and subchronic toxicity tests with rodents, in order to assess potential risks posed to terrestrial wildlife. Dosages were chosen to bracket worst-case environmental exposure scenarios. In acute tests, adult female Wistar rats were given single po dosages of naphthenic acids at either 3, 30, or 300 mg per kg body weight (mg/kg), while adult male rats received 300 mg/kg. Food consumption was temporarily suppressed in the high-dose groups of both sexes. Following euthanasia 14 days later, histopathology revealed a significant incidence of pericholangitis in the high-dose group of both sexes, suggesting hepatotoxicity as an acute effect. Other histological lesions included brain hemorrhage in high-dose males, and cardiac periarteriolar necrosis and fibrosis in female rats. In subchronic tests, naphthenic acids were po administered to female Wistar rats at 0.6, 6, or 60 mg/kg, 5 days per week for 90 days. Results again suggested the liver as a potential target organ. The relative liver weight in the high-dose group was 35% higher than in controls. Biochemical analysis revealed elevated blood amylase (30% above controls) and hypocholesterolemia (43% below controls) in high-dose rats. Excessive hepatic glycogen accumulation was observed in 42% of animals in this group. These results indicate that, under worst-case exposure conditions, acute toxicity is unlikely in wild mammals exposed to naphthenic acids in AOS tailings pond water, but repeated exposure may have adverse health effects.     

Oxidation of retinoic acids in hepatic microsomes of wild bullfrogs Lithobates catesbeianus environmentally-exposed to a gradient of agricultural contamination

Agricultural contaminants are suspected of contributing to the increased incidence of deformities and the decline of amphibians populations worldwide. Many authors have further suggested that a retinoid effect could be implicated in teratogenic mechanisms since the reported deformities resemble those caused by abnormal levels of retinoic acid (RA). We previously reported altered retinoid concentrations in male bullfrogs from the Yamaska River basin (Québec, Canada) associated with moderate-to-high agricultural activity, and the findings were consistent with a possible effect on hepatic RA oxidation. An in vitro assay was therefore optimized and hepatic microsomal RA oxidation in bullfrogs was found to be quite different from that of other vertebrates. With either all-transRA (atRA) or 13cisRA as the substrate, the major metabolite generated was at4-oxo-RA. The reaction with 13cisRA as substrate, markedly greater compared with atRA, was enhanced in the presence of a reducing agent and inhibited by cytochrome P450 inhibitors in a dose-dependent manner. Hepatic RA oxidation in male bullfrogs showed significant differences between sites with no clear relationship to a gradient of agricultural activity or 13cis-4-oxo-RA quantified in plasma. In contrast, the in vitro RA oxidation in females increased with the levels of contamination and coincided in vivo with higher plasma 13cis-4-oxo-RA concentration. The levels of circulating 4-oxo-derivatives could be influenced by hepatic RA oxidative metabolism as well as isomerization conditions or RA precursor levels.     

Toxicity of ZnO nanoparticles (NPs) to THP-1 macrophages: interactions with saturated or unsaturated free fatty acids

In a biological microenvironment, free fatty acids (FFA) as ubiquitous biological molecules might interact with nanoparticles (NPs) and consequently change the toxicological responses. However, whether the chemical structures of FFA could influence their interactions with NPs remain unknown. This study investigated the interactions between ZnO NPs and saturated or unsaturated FFA (complexed to BSA), namely stearic acid (SA, C18:0), oleic acid (OA, C18:1), and α-linolenic acid (ALA, C18:3). It was shown that BSA, SA, OA, and ALA increased the atomic force microscope (AFM) heights as well the polydispersity index (PDI) of ZnO NPs. BSA modestly protected THP-1 macrophages from ZnO NP exposure, whereas OA and ALA led to relatively less cyto-protective effects of BSA. Moreover, only co-exposure to ZnO NPs and SA significantly promoted the release of interleukin-8. BSA, SA, OA, and ALA equally changed intracellular ROS and Zn ions associated with ZnO exposure, but co-exposure to ZnO NPs and OA/ALA particularly activated the expression of endoplasmic reticulum stress-apoptosis genes. In combination, these results showed that FFA could influence the colloidal aspects and toxicological signaling pathway of ZnO NPs, which is dependent on the number of unsaturated bonds of FFA.     

Mortality of embryos,developmental disorders and changes in biochemical parameters in marsh frog (Rana ridibunda) tadpoles exposed to the water-soluble fraction of Kazakhstan crude oil and O-Xylene

The effects of different concentrations of water-soluble fraction of crude oil (WSFO) from the Zhanazhol oil field (Aktobe region, Kazakhstan) and compared to o-xylene, prevalent in this oil, on growth and development of marsh frog (Rana ridibunda) were assessed. In subchronic experiments (7 d), a dose-related increase in mortality and incidence of deformities in embryos were observed. In chronic experiments (60 d; starting from the Gosner stage 26), a dose-dependent decrease in body weight, size and developmental delay by 3–4 stages were also detected. In addition, the content of lipid hyperoxide (LHO) and malondialdehyde (MDA), as well as activities of superoxide dismutase (SOD) and catalase (CAT) enzymes in liver of the tadpoles were determined at the end of chronic experiment. Exposure to 0.5 mg/L or 1.5 mg/L WSFO elevated the content of LHO by 76% and 86%, and MDA by 47% and 58% but decreased SOD activity by 26% and 49%, and CAT by 35% and 46%, respectively. A less pronounced adverse effect was found after chronic exposure to the same concentrations of o-xylene. In tadpole liver exposed to o-xylene levels of LHO was increased by 40% and 51%, MDA by 11% and 29%, while the activity of SOD was lowered by 18% and 41%, and CAT – by 13% and 37% in the 0.5 mg/L and 1.5 mg/L treatment groups, respectively. Data demonstrated the embryotoxic and teratogenic effects attributed to WSFO and o-xylene exposure which may involve oxidative stress mechanisms.