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1.
Due to hydrophobic and persistent properties, polycyclic aromatic hydrocarbons (PAHs) have a high capacity to accumulate in sediment. Sediment quality criteria, for the assessment of habitat quality and risk for aquatic life, include understanding the fate and effects of PAHs. In the context of European regulation (REACH and Water Framework Directive), the first objective was to assess the influence of sediment composition on the toxicity of two model PAHs, benzo[a]pyrene and fluoranthene using 10-day zebrafish embryo-larval assay. This procedure was undertaken with an artificial sediment in order to limit natural sediment variability. A suitable sediment composition might be then validated for zebrafish and proposed in a new OECD guideline for chemicals testing. Second, a comparative study of toxicity responses from this exposure protocol was then performed using another OECD species, the Japanese medaka. The potential toxicity of both PAHs was assessed through lethal (e.g., survival, hatching success) and sublethal endpoints (e.g., abnormalities, PMR, and EROD) measured at different developmental stages, adapted to the embryonic development time of both species. Regarding effects observed for both species, a suitable artificial sediment composition for PAH toxicity testing was set at 92.5 % dry weight (dw) silica of 0.2–0.5-mm grain size, 5 % dw kaolin clay without organic matter for zebrafish, and 2.5 % dw blond peat in more only for Japanese medaka. PAH bioavailability and toxicity were highly dependent on the fraction of organic matter in sediment and of the K ow coefficients of the tested compounds. The biological responses observed were also dependent of the species under consideration. Japanese medaka embryos appeared more robust than zebrafish embryos for understanding the toxicity of PAHs following a sediment contact test, due to the longer exposure duration and lower sensitivity of sediment physical properties.  相似文献   

2.
A natural sediment spiked with three individual polycyclic aromatic hydrocarbons (PAHs; pyrene, phenanthrene and benzo[a]pyrene) was used to expose zebrafish embryos and larvae during 4 days. The total PAH concentration was 4.4 μg g?1 which is in the range found in sediment from contaminated areas. Quantification of metabolites in the larvae after exposure confirmed the actual contamination of the larvae and indicated an active metabolism especially for pyrene and benzo[a]pyrene. After a transfer in a clean medium, the larvae were reared to adulthood and evaluated for survival, growth, reproduction, and behavior. Measured endpoints revealed a late disruption of growth (appearing at 5 months) and a trend toward a lower reproductive ability. Adults of embryos exposed to sediment spiked with PAHs displayed lethargic and/or anxiety-like behaviors. This latter behavior was also identified in offspring at larval stage. All together, these effects could have detrimental consequences on fish performances and contribution to recruitment.  相似文献   

3.
In aquatic environments, polycyclic aromatic hydrocarbons (PAHs) mostly occur as complex mixtures, for which risk assessment remains problematic. To better understand the effects of PAH mixture toxicity on fish early life stages, this study compared the developmental toxicity of three PAH complex mixtures. These mixtures were extracted from a PAH-contaminated sediment (Seine estuary, France) and two oils (Arabian Light and Erika). For each fraction, artificial sediment was spiked at three different environmental concentrations roughly equivalent to 0.5, 4, and 10 μg total PAH g?1 dw. Japanese medaka embryos were incubated on these PAH-spiked sediments throughout their development, right up until hatching. Several endpoints were recorded at different developmental stages, including acute endpoints, morphological abnormalities, larvae locomotion, and genotoxicity (comet and micronucleus assays). The three PAH fractions delayed hatching, induced developmental abnormalities, disrupted larvae swimming activity, and damaged DNA at environmental concentrations. Differences in toxicity levels, likely related to differences in PAH proportions, were highlighted between fractions. The Arabian Light and Erika petrogenic fractions, containing a high proportion of alkylated PAHs and low molecular weight PAHs, were more toxic to Japanese medaka early life stages than the pyrolytic fraction. This was not supported by the toxic equivalency approach, which appeared unsuitable for assessing the toxicity of the three PAH fractions to fish early life stages. This study highlights the potential risks posed by environmental mixtures of alkylated and low molecular weight PAHs to early stages of fish development.  相似文献   

4.
A new gravel-contact assay using rainbow trout, Oncorhynchus mykiss, embryos was developed to assess the toxicity of polycyclic aromatic hydrocarbons (PAHs) and other hydrophobic compounds. Environmentally realistic exposure conditions were mimicked with a direct exposure of eyed rainbow trout embryos incubated onto chemical-spiked gravels until hatching at 10 °C. Several endpoints were recorded including survival, hatching delay, hatching success, biometry, developmental abnormalities, and DNA damage (comet and micronucleus assays). This bioassay was firstly tested with two model PAHs, fluoranthene and benzo[a]pyrene. Then, the method was applied to compare the toxicity of three PAH complex mixtures characterized by different PAH compositions: a pyrolytic extract from a PAH-contaminated sediment (Seine estuary, France) and two petrogenic extracts from Arabian Light and Erika oils, at two environmental concentrations, 3 and 10 μg g?1 sum of PAHs. The degree and spectrum of toxicity were different according to the extract considered. Acute effects including embryo mortality and decreased hatching success were observed only for Erika oil extract. Arabian Light and pyrolytic extracts induced mainly sublethal effects including reduced larvae size and hemorrhages. Arabian Light and Erika extracts both induced repairable DNA damage as revealed by the comet assay versus the micronucleus assay. The concentration and proportion of methylphenanthrenes and methylanthracenes appeared to drive the toxicity of the three PAH fractions tested, featuring a toxic gradient as follows: pyrolytic?Arabian Light?Erika. The minimal concentration causing developmental defects was as low as 0.7 μg g?1 sum of PAHs, indicating the high sensitivity of the assay and validating its use for toxicity assessment of particle-bound pollutants.  相似文献   

5.
Superficial sediments collected from seven estuarine systems located along the Portuguese coast were analyzed for 7 polychlorinated dibenzo-p-dioxins (PCDDs), 10 polychlorinated dibenzofurans (PCDFs), and 12 dioxin-like polychlorinated biphenyls (dl-PCBs). Total PCDD/F concentration ranged from 4.6 to 464 pg g?1 dry weight (dw), while that of dl-PCBs varied from 26.6 to 8,693 pg g?1 dw. In general, the highest PCDD/F and dl-PCB concentrations were associated with densely populated and industrially impacted areas. Additionally, PCDD/F revealed a predominance of octachlorodibenzodioxin (OCDD) to total PCDD/Fs, while PCB 118 was the major contributor to total dl-PCBs. This study provided a global perspective of the contamination status of Portuguese estuaries by dioxin-like compounds and allowed a comparison between the investigated systems and other systems worldwide. PCDD/F and dl-PCB levels found in the collected sediments were lower than those of highly impacted areas from different parts of the globe. Nevertheless, comparison with guidelines and quality standards from other countries indicated that some Portuguese estuarine areas with a high industrialization level present PCDD/F and dl-PCB concentrations in superficial sediment that may constitute a risk to aquatic organisms.  相似文献   

6.
Organochlorine pesticides (OCPs) are ubiquitous pollutants, and their presence in urban lakes is a concern for human and ecological health. Surface sediments in the East Lake, China, were collected in winter 2012 and summer 2013 to investigate concentrations, distribution patterns, possible sources, and potential ecological risks of OCPs in this area. The total concentrations of 14 OCPs ranged from 6.3 to 400 ng g?1 dry weight (dw) with an average concentration of 79 ng g?1 dw. The mean values of hexachlorocyclohexanes (HCHs) (α-, β-, γ-, and δ-HCH) and dichlorodiphenyltrichloroethanes (DDTs) (p,p’-DDE, p,p’-DDD, and p,p’-DDT) were 36 and 7.6 ng g?1 dw, accounting for 45 and 10 % of the total OCPs, respectively. The concentrations of OCPs in sediment samples collected in winter were significantly higher than those in summer, especially the HCHs, of which in winter were two times greater than summer. Composition analyses indicated that DDTs and endosulfan were mainly from historical contribution. Historical use of technical HCH and new input of lindane were probably the source of HCHs in the East Lake. Most sampling sites of HCHs and DDTs were found to have the potential ecological risk based on levels specified in the sediment quality standards.  相似文献   

7.
Toxicity evaluation is an important segment in sediment quality monitoring in order to protect aquatic organisms and human health. The purpose of this study is to assess the toxicity of sediments from three sediment cores in Yangtze River Estuary, China, using the zebrafish (Danio rerio) embryo tests. Fertilized zebrafish eggs were exposed to both whole sediments and sediment organic extracts prepared from collected sediments, in order to provide a comprehensive and realistic insight into the bioavailable toxicity potential of the sediments. As end points, development parameters (mortality, hatching rate, and abnormality) in the developing embryos were recorded during the 96-h exposure. The results showed that some samples increased mortality, inhibited the hatching of embryos, and induced morphological abnormalities. The embryonic toxicities presented serrated changes and irregular distribution with depth, which may be related to hydrodynamic effect and unstable environmental input. However, lethal and sub-lethal effects were more significant at the sub-surface sediments (10~40 cm), which indicated that the pollution is more serious in recent decades.  相似文献   

8.
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10.
There is a consensus within the scientific community that sediments act as a long-term sink for a variety of organic and inorganic pollutants, which, however, can re-enter the water column upon resuspension of deposited material under certain hydraulic conditions such as flood events. Within the implementation of the European Water Framework Directive, it is important to understand the potential short- and long-term impact of suspended particulate matter (SPM)-associated contaminants on aquatic organisms as well as the related uptake mechanisms for a sound risk assessment. To elucidate the effects of sediment-bound organic pollutants, such as polycyclic aromatic hydrocarbons (PAHs), rainbow trout (Oncorhynchus mykiss) were exposed to three resuspended natural sediments with different contamination levels. Physicochemical parameters including dissolved oxygen concentration, pH and temperature, total PAH concentration in sediments and SPM as well as different biomarkers of exposure in fish (7-ethoxyresorufin O-deethylase activity, biliary PAH metabolites, micronuclei, and lipid peroxidation) were measured following seven days of exposure within an annular flume, a device to assess erosion and deposition processes of cohesive sediment. Concentrations of PAHs in SPM remained constant and represented the different contamination levels in the un-suspended sediments. Significant differences in bile metabolite concentrations as well as in 7-ethoxyresorufin O-deethylase induction compared to control experiments (untreated animals and animals that were exposed in the annular flume without sediment) were observed for all exposure scenarios. The ratio between 1-hydroxypyrene in bile from fish exposed to the three different contamination levels was 1.0:3.6:10.7 and correlated well with (1) the ratio of pyrene concentrations in corresponding sediments which was 1.0:3.1:12.7 and (2) with the ratio of particle-bound pyrene in SPM which was 1.0:2.7:11.7. In contrast, hepatic lipid peroxidation and micronuclei formation represented the different contamination levels less conclusive. The results of this study clearly demonstrate that firmly bound PAH from aged sediments can become bioaccessible upon resuspension under flood-like conditions and are readily absorbed by aquatic organisms such as rainbow trout. Associated short-term effects were clearly documented and possible adverse long-term impacts due to genotoxicity are likely to follow.  相似文献   

11.
Within the framework of the MYTIOR project in 2009, heavy metals and organic compounds contaminations were assessed in transplanted mussels in 16 different stations along the coasts of Libya. These stations were located at miles offshore industrial/urban sources but in open sea providing original results related to the background contamination rather than linked to a specific coastal source of pollutants. Results indicated mercury (Hg, 0.045–0.066 mg/kg dry weight (dw)), lead (Pb, 0.44–0, 71 mg/kg dw) and copper (Cu, 3.56–4.21 mg/kg dw) were in the same range or at lower value than control for all stations. Chromium (Cr) in Meleta (3.08 mg/kg dw) and Bomba (3.80 mg/kg dw) and Cadmium values in all stations (1.21–2.41 mg/kg dw) were above control. Meleta, stations from the gulf of Syrt and the three eastern stations were the most affected stations by nickel (max at 5.83 mg/kg dw in Syrt) when zinc was in the same range (141–197 mg/kg dw) and above the control (92 mg/kg dw) at all stations. Polycyclic aromatic hydrocarbon (PAH) levels were found in the range of 16.8–42.8 mg/kg (dry weight) indicating low levels along the Libyan coast with acenaphthene and benzo (a, b, k) pyrenes detected mainly in western Libya. The study of PAH ratios indicated a mixed petrogenic/pyrolytic origin. The only polychlorinated biphenyls (PCBs) found in Libya were PCB 101 in one location and PCB 153 in Tripoli, Garrapoli, Syrt, Ras Lanuf and Benghazi (1.2–1.9 μg/kg dw). Insecticides were lower than control in all stations except DDT, only detected in Misratah (3.5 μg/kg dw). Overall, the results indicated a low background contamination and a low pollution extent according to the environmental pressure occurring offshore the Libyan coast.  相似文献   

12.
Concentrations and profiles of 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) were investigated in sediment and plants collected from a salt marsh in the Tejo estuary, Portugal. The highest PCDD/F and dl-PCB concentrations were detected in uncolonized sediments, averaging 325.25?±?57.55 pg g?1 dry weight (dw) and 8,146.33?±?2,142.14 pg g?1 dw, respectively. The plants Sarcocornia perennis and Halimione portulacoides growing in PCDD/F and dl-PCB contaminated sediments accumulated contaminants in roots, stems, and leaves. It was observed that PCDD/F and dl-PCB concentrations in roots were significantly lower in comparison with stems and leaves. In general, concentration of ΣPCDD/Fs and Σdl-PCBs in H. portulacoides tissues were found to be twofold higher than those in S. perennis, indicating a difference in the accumulation capability of both species. Furthermore, congener profiles changed between sediments and plant tissues, reflecting a selective accumulation of low chlorinated PCDD/Fs and non-ortho dl-PCBs in plants.  相似文献   

13.
The goal was to determine dissolution potency of betulinol and wood sterols (WSs) from pulp and paper mill-contaminated sediments and the current stratification for assessment the load due to potential erosion in the river-like watercourse. Both compounds are wood extractives, which may be toxic to benthos and fish. This research continues a study in which other wood extractives, resin acids and their derivative, retene, were analysed. Sediments were collected from 1, 3.5, 12, 15, and 33 km downstream from the pulp and paper mills, and from 2 upstream reference sites. The dissolution potency into sediment–water elutriates (1?+?4?v/v) was studied by two agitation times and temperatures. The vertical amounts of extractives were determined from the uppermost 20 cm of sediment. The amounts of extractives potentially released were estimated from the sediment layers 0–2 and 2–5 cm by using spatial interpolation. According to the interpolation, the total amount of betulinol and β-sitosterol was calculated as kg/ha in the whole sediment area. Significant concentrations of betulinol (1,666 μg/g, dw) and WSs (2,886 μg/g, dw) were measured from the sediments. According spatial interpolation, the highest calculated amount of betulinol (4,726 kg/ha) and that of the most abundant WS, β-sitosterol (3,571 kg/ha), were in the lake where the effluents were discharged. In the dissolution experiment, the highest concentration of betulinol in sediment (0–10 cm) and elutriate was 412 μg/g (dw) and 165 μg/l, respectively. For WSs, concentrations were 768 μg/g (dw) in sediment and 273 μg/l in elutriate. In a worst-case scenario, betulinol may be desorbed to water in concentrations which are hazardous to aquatic animals. Instead WSs are not a risk in this study area. The amount of desorption varied depending on the concentration of contaminants in sediment, the nature of disturbance, and the sediment organic carbon content.  相似文献   

14.
Degradation of three sulfonamides (SAs), namely sulfamethoxazole (SMX), sulfamethazine (SMZ), and sulfadimethoxine (SDM) in surface water and sediments collected from Taihu Lake and Dianchi Lake, China was investigated in this study. The surface water (5–10 cm) was collected from the east region of Taihu Lake, China. Two sets of degradation experiments were conducted in 3-L glass bottles containing 2 L of fresh lake water and 100 μg/L of individual SAs aerated by bubbling air at a rate of approximately 1.2 L/min, one of which was sterilized by the addition of NaN3 (0.1 %). Sediment samples were taken from Taihu Lake and Dianchi Lake, China. For the sediment experiment, 5 g of sediment were weighed into a 50-mL glass tube, with 10 mg/kg of individual SAs. Different experimental conditions including the sediment types, sterilization, light exposure, and redox condition were also considered in the experiments. The three SAs degraded in lake water with half-lives (t 1/2) of 10.5–12.9 days, and the half-lives increased significantly to 31.9–49.8 days in the sterilized water. SMZ and SDM were degraded by abiotic processes in Taihu and Dianchi sediments, and the different experimental conditions and sediments characteristics had no significant effect on their declines. SMX, however, was mainly transformed by facultative anaerobes in Taihu and Dianchi sediments under anaerobic conditions, and the degradation rate of SMX in non-sterile sediment (t 1/2 of 9.6–16.7 days) were higher than in sterilized sediment (t 1/2 of 18.7–135.9 days). Under abiotic conditions, degradation of SMX in Dianchi sediment was faster than in Taihu sediment, probably due to the higher organic matter content and inorganic photosensitizers concentrations in Dianchi sediment. High initial SAs concentration inhibited the SAs degradation, which was likely related to the inhibition of microorganism activities by high SAs levels in sediments. Results from this study could provide information on the persistence of commonly used sulfanomides antibiotics in lake environment.  相似文献   

15.
Polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) were determined in 18 surface sediment samples collected from Bizerte lagoon, Tunisia. The total concentrations of ten PCBs (∑PCBs) and of four OCPs (∑OCPs) in the sediments from this area ranged from 0.8 to 14.6 ng g?1 dw (average value, 3.9 ng g?1 dw) and from 1.1 to 14.0 ng g?1 dw (average value, 3.3 ng g?1 dw), respectively. Among the OCPs, the range of concentrations of dichlorodiphenyltrichloroethane and its metabolites (DDTs) and hexachlorobenzene (HCB) were 0.3–11.5 ng g?1 dw (1.9 ng g?1 dw) and 0.6–2.5 ng g?1 dw (1.4 ng g?1 dw), respectively. Compositional analyses of the POPs indicated that PCB 153, 138 and 180 were the predominant congeners accounting for 60 % of the total PCBs. In addition, p,p′-DDT was found to be the dominant DDTs, demonstrating recent inputs in the environment. Compared with some other regions of the world, the Bizerte lagoon exhibited low levels of PCBs and moderate levels of HCB and DDTs. The high ratios ΣPCBs/ΣDDTs indicated predominant industrial versus agricultural activities in this area. According to the established guidelines for sediment quality, the risk of adverse biological effects from such levels of OCPs and PCBs, as recorded at most of the study sites, was insignificant. However, the higher concentrations in stations S1 and S3 could cause biological damage.  相似文献   

16.
This study investigated the contamination levels, homologue, and congener profiles and evaluated the residue inventory of polychlorinated biphenyls (PCBs) in sediment of the Songhua River in the vicinity of cement plant. The total concentration of detected 35 PCB congeners ranged from 1.12 to 2.19 ng/g dry weight (dw) in sediment, with a mean value of 1.56 ng/g dw, and the concentration of PCBs decreased in the following order: the downstream?>?cement plant?>?upstream. The results of total organic carbon (TOC)-normalized PCB concentrations indicate that the sediments have low potential ecological risk in this area. The study results on homologue and congener of PCBs show that the emissions from cement production could be the major sources of PCBs in sediment, and the low-chlorinated PCBs will be exchanged among air, water, and sediment with increasing temperature during summer and flow to downstream with water during the wet season. The spatial density and total burden of PCBs in the surface sediments were 17.2 ng/cm2 and 1.2 kg, respectively. To our knowledge, this study is the first to explore the pollution characteristics of unintentionally produced PCB emissions from cement industry by means of monitoring sediment samples.  相似文献   

17.
The concentrations, compositional profiles, and possible sources of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/PCDF) in water column and surface sediment samples from the lower reaches of the Yangtze River were investigated, and the potential risks posed by these chemicals were evaluated. Concentrations of the 17 2,3,7,8-substituted PCDD/PCDF in the water and sediment samples ranged from 0.45 to 10.0 pg TEQ/L (mean 2.34 pg TEQ/L) and from 0.31 to 51 pg TEQ/g dw (mean 5.25 pg TEQ/g dw), respectively. Water and sediments from the Nanjing–Nantong sections of the Yangtze River were found to have elevated PCDD/PCDF levels, and the PCDD/PCDF TEQ concentrations in 13 sediment samples exceeded the probable effect levels (PEL), above which adverse biological effects are likely. Multivariate statistical analysis indicated that the principal PCDD/PCDF contamination source for the water and sediment was the production and use of pentachlorophenol (PCP) or sodium pentachlorophenate (PCP-Na). PCDD/PCDF concentrations were stable over time near the Yangtze River Estuary but had increased sharply in recent decades in the Nanjing–Nantong section, which will have been related to the growth in industrial activities and other anthropogenic PCDD/PCDF sources. Total organic carbon (TOC) and PCDD/PCDF concentrations correlated significantly (R?=?0.474, P?<?0.01), suggesting that TOC plays an important role in the transport and re-distribution of PCDD/PCDF in the Yangtze River basin.  相似文献   

18.
Seventeen organochlorine pesticides (OCPs) were investigated in the water and sediments from a waterbird-inhabited lake (Yangchaihu Lake) to evaluate their current pollution levels and potential risks. The concentrations of total OCPs in water and sediments were 10.12–59.75 ng/l and 4.25–27.35 ng/g dry weight, respectively. Hexachlorocyclohexanes (HCHs) and dichlorodiphenyltrichloroethanes (DDTs) were the most abundant OCPs, while HCB and cyclodiene pesticides were detected with low levels. Levels of ∑OCPs (sum of 17 OCPs) at sites highly influenced by waterbirds were significantly higher than the sites with no significant waterbird populations (one-way ANOVA, P?相似文献   

19.
Diphenyl ether and its derivatives are widely used in the industry of spices, dyes, agrochemicals, and pharmaceuticals. Following the previous study, we selected 4,4′-dihydroxydiphenyl ether, 4,4′-difluorodiphenyl ether, 4,4′-dichlorodiphenyl ether, and 4,4′-dibromodiphenyl ether as research objects. The LC50 (96 h) values for these compounds in adult zebrafish were determined with the acute test. Also, developmental toxicities of the four substances to zebrafish embryos were observed at 24, 48, 72, and 96 hpf. All the LC50 (96 h) values of these compounds were between 1 and 10 mg/L, suggesting that they all had moderate toxicity to adult zebrafish. The embryonic test demonstrated that with increasing doses, 4,4′-dihydroxydiphenyl ether decreased the hatching rate, while 4,4′-difluorodiphenyl ether, 4,4′-dichlorodiphenyl ether, and 4,4′-dibromodiphenyl ether delayed the hatching time but had little effect on final hatchability at 96 hpf. All of these compounds inhibited larval growth, especially 4,4′-dihydroxydiphenyl ether. Exposure to these chemicals induced embryo yolk sac and pericardial edema. Spine deformation was visible in hatched larvae after 96 hpf 4,4′-dihydroxydiphenyl ether exposure, while tail curvature was observed for the halogenated compounds. The overall results indicated that 4,4′-dihydroxydiphenyl ether, 4,4′-difluorodiphenyl ether, 4,4′-dichlorodiphenyl ether, and 4,4′-dibromodiphenyl ether all had significant toxicity on adult and embryonic zebrafish.  相似文献   

20.
Using rare minnow (Gobiocypris rarus) at early-life stages as experimental models, the developmental toxicity of five widely used triazole fungicides (myclobutanil, fluconazole, flusilazole, triflumizole, and epoxiconazole) were investigated following exposure to 1–15 mg/L for 72 h. Meanwhile, morphological parameters (body length, body weight, and heart rate), enzyme activities (superoxide dismutase (SOD), glutathione S-transferase (GST), adenosine triphosphatase (ATPase), and acetyl cholinesterase (AChE)), and mRNA levels (hsp70, mstn, mt, apaf1, vezf1, and cyp1a) were also recorded following exposure to 0.2, 1.0, and 5.0 mg/L for 72 h. Results indicated that increased malformation and mortality, decreased body length, body weight, and heart rate provide a concentration-dependent pattern; values of 72 h LC50 (median lethal concentration) and EC50 (median effective concentration) ranged from 3 to 12 mg/L. Most importantly, the results of the present study suggest that even at the lowest concentration, 0.2 mg/L, five triazole fungicides also caused notable changes in enzyme activities and mRNA levels. Overall, the present study points out that those five triazole fungicides are highly toxic to the early development of G. rarus embryos. The information presented in this study will be helpful in better understanding the toxicity induced by triazole fungicides in fish embryos.  相似文献   

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