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1.
Lethal effects of active ingredients and formulations of widely used soybean pesticides were assessed with the Hydra attenuata toxicity test. Studied pesticides were insecticides chlorpyrifos and cypermethrin, and herbicide glyphosate. Results indicate the following toxicity trend: chlorpyrifos > cypermethrin > glyphosate. Tested active ingredients of insecticides and respective formulations did not significantly differ between them. Glyphosate formulation exhibited higher toxicity at low concentrations (LC1–10) respect to active ingredient, reversing this behavior at higher concentrations (LC50–90). Comparing H. attenuata sensitivity with existent toxicity data for aquatic organisms indicates that this species is poorly sensitive to tested insecticides and highly sensitive to the herbicide.  相似文献   

2.
Many chemicals, including fertilizers, herbicides, and insecticides, are routinely applied to turf in the care and maintenance of lawns. These chemicals have the potential to leach into nearby surface waters and adversely affect aquatic biota. In this study, we evaluated the lethal and genotoxic effects of chemicals used in lawn care on an early life stage of freshwater mussels (Utterbackia imbecillis). The chemicals tested were copper and commercial formulations of atrazine, glyphosate, carbaryl, and diazinon. Mussel glochidia were exposed to chemicals singly or in combination (equitoxic and environmentally realistic mixtures) for 24 h and toxic interactions were evaluated with Markings additive index. Genotoxicity was quantified with the alkaline single-cell gel electrophoresis assay (Comet assay). In acute tests, copper was the most toxic of all chemicals evaluated (LC50 = 37.4 µg/L) and carbaryl was the most toxic of all pesticides evaluated (LC50 = 7.9 mg/L). In comparison to other aquatic organisms commonly used in toxicity tests (e.g., amphipods, cladocerans, and chironomids), mussel glochidia were as or more sensitive to the chemicals evaluated with the exception of diazinon, where mussels were observed to be less sensitive. The combined toxicity of equitoxic and environmentally realistic mixtures to mussels was additive. Genotoxic responses were observed in mussels exposed to copper, atrazine and diazinon at levels below their respective no-observed-effect concentrations. Together, these data indicate that freshwater mussels are among the most sensitive aquatic organisms tested for some chemicals commonly used in lawn care and that DNA damage may be useful as a screening tool to evaluate potential sublethal effects of lawn care products on non-target aquatic organisms.  相似文献   

3.
When assessing pesticides toxicity on test organisms, the use of organic solvents is frequently required to formulate solutions of pesticides having low or moderate water solubility. These solvents may influence the results of toxicity tests. This study examined the effects of ethanol and dimethyl sulfoxide (DMSO) on two species of algae, Chlorella vulgaris and Selenastrum capricornutum. The interactives effects of these organic solvents with various concentrations of atrazine were tested to analyse how these solvents can affect the toxicity of atrazine. Toxicity was measured as the change in chlorophyll (a) content (estimated via fluorescence) in cultures of the test organism that were incubated over a 96-h period at standard conditions of temperature and light. In the absence of atrazine, ethanol was toxic to both algae, yielding significant inhibition of chlorophyll (a) content at concentrations as low as 0.05%. Morever, S. capricornutum was less sensitive to ethanol than was C. vulgaris. DMSO did not show any toxic effects on either chlorophycea. At concentrations up to 0.5%, DMSO interacted additively with atrazine for both chlorophycea. An additive response was also observed with ethanol towards S. capricornutum. For C. vulgaris, ethanol interacted antagonistically at most of atrazine concentrations and gave few additive and synergistic interactions. An additive response of an atrazine-solvent mixture indicate the inherent toxicity of the atrazine. Since DMSO generally gave additive responses for both algae, it can be considered as an adequate organic solvent to use in bioassays.  相似文献   

4.

The lanthanides are considered emerging contaminants but information on their long-term toxicity to aquatic species under environmentally relevant conditions is scarce. We aimed to fill this gap by evaluating the long-term adverse effects of gadolinium on the freshwater model—crustacean Daphnia magna. The exposure of D. magna for up to 39 days to 0.1 mg Gd/L (a 21-days chronic toxicity NOEC value derived by us formerly) in the lake water had no negative effect (p?>?0.05) on vitality, size and reproduction of parent animals as well as their offspring. Thus, assumingly the current Gd contamination levels of surface waters pose no hazard to aquatic crustaceans that in general are very sensitive to various pollutants. Moreover, presence of 0.1 mg Gd/L in the lake water even mitigated the long-term toxic effect of 0.2 mg Ni/L (studied as a model co-contaminant) to D. magna’s vitality and productivity.

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5.
The toxicity assessment of chemicals is one of the main issues in the current policies in order to protect the health of the environment and human beings. Food and cosmetic additives have been extensively studied in relation to their toxicity to humans, but data about their ecotoxicological effects are scarce. The aim of this study was to evaluate the toxic effects of the additive 6-methylcoumarine in the aquatic milieu using a test battery comprising experimental model systems from different trophic levels. The inhibition of bioluminiscence was studied in the bacteria Vibrio fischeri (decomposer), the inhibition of growth was evaluated in the alga Chlorella vulgaris (producer) and immobilization was studied in the cladoceran Daphnia magna (first consumer). Finally, several end points were evaluated in the RTG-2 salmonid fish cell line, including neutral red uptake, protein content, methylthiazol tetrazolium salt metabolization, glucose-6-phosphate dehydrogenase activity, lactate dehydrogenase activity and leakage, and morphology. The sensitivity of the test systems employed was as follows: V. fischeri > D. magna > C. vulgaris > RTG-2 cell line. The results show that 6-methylcoumarine is not expected to produce acute toxic effects on the aquatic biota. However, chronic and synergistic effects with other chemicals cannot be excluded and should be further investigated.  相似文献   

6.
The acute toxicity of terbufos and its major metabolites, tested alone, in binary mixtures or in combination with atrazine were evaluated using neonates of the cladoceran Ceriodaphnia cf dubia. Terbufos, terbufos sulfoxide, and terbufos sulfone tested individually were highly toxic to C. cf dubia, with mean 96-h EC50 values of 0.08, 0.36, and 0.19 μg/l, respectively. The addition of atrazine (10 μg/l) significantly increased the toxicity of terbufos. The toxicity of terbufos sulfone was unaffected by atrazine, whereas the results for terbufos sulfoxide were equivocal. Equitoxic mixtures of the metabolites showed additive toxicity to C. cf dubia. The high toxicities of terbufos and its environmentally persistent oxidative metabolites suggest that contamination of aquatic systems with this insecticide mixture and the coapplied herbicide atrazine might pose a greater hazard to some biota than their individual toxicities.  相似文献   

7.
Boron is an essential nutrient for plants and an essential element for many organisms, but can be toxic to aquatic and terrestrial organisms above certain concentrations. The aim of this research was to determine boron concentrations in water, sediment and biotic samples (Gammaridae spp.-Crustacea, Helix sp.-Gastropoda, Donax sp.-Bivalvia, Helobdella sp.-Hirudinae, Ephemeroptera nymph, Chrinomidae larvae, Tipulidae larvae-Insecta, Rana sp.-Amphibia, Natrix sp.-Serpentes, fish sample Leiscus cephalus (Linnaeus, 1758) and leaves of Salix sp.-Salicacea from Seydi Stream (Kırka-Eskişehir). Our results have shown that boron concentrations of the Seydi Stream water is higher than the Turkish Environmental Guidelines standard (>1 mg L−1) and in Europe (mean values typically below 0.6 mg L−1).  相似文献   

8.
The use of herbicides for agricultural and aquatic weed control has increased worldwide. These substances are potentially toxic pollutants because they induce the production of reactive oxygen species for biological systems and exert oxidative stress in nontarget organisms living in the treated aquatic systems. Recent evidence suggests differences in the toxicity of glyphosate in the form of an active ingredient compared to the toxicity of glyphosate in combination with surfactants, such as those found in commercial formulations. In Mexico, one of the most widely used glyphosate-based herbicides is Yerbimat, which has agricultural as well as aquatic weed control applications. However, there are no aquatic toxicity data, particularly regarding native fish. Therefore, we determined the acute toxicity of commercial-formulation Yerbimat in a static bioassay at 96 h (LC50). We also determined its toxicity at 96 h in sublethal concentrations to assess the lipid peroxidation levels (LPX), catalase activity, hepatic glycogen content, and histological damage in the liver and gills of the fish Goodea atripinnis associated with chronic exposure (75 days). The LC50 was 38.95 ± 0.33 mg/L. The results of the short-term exposure study indicate that Yerbimat can potentially induce oxidative stress in G. atripinnis, because LPX was increased in the gills and liver. Catalase activity was reduced in the gills but increased in the liver, whereas hepatic glycogen was depleted. Chronic exposure was associated with histopathological damage in the gills and liver, some of which was irreversible. Yerbimat represents a potential risk for aquatic biota; therefore, we recommend that its application be carefully considered.  相似文献   

9.
Organophosphorus (OP) insecticides have high acute toxicity toward many nontarget vertebrate and invertebrate organisms, but direct measurement of OPs in environmental samples is difficult because their concentrations may fall below detection limits within hours to days after entering aquatic ecosystems. Because OPs exert toxicity through cholinesterase (ChE) inhibition, which may persist for up to several weeks, ChE inhibition has been widely used in aquatic ecosystems as a biomarker for OP exposure in aquatic organisms. However, the biological, environmental, and methodological factors affecting ChE activity have not been well documented and must be considered and understood before ChE activity can be used as a dependable indicator of OP exposure to aquatic organisms. This study examined the influence of water temperature, size of larval and juvenile walleye (Stizostedion vitreum), stress, long-term storage, postmortem changes, and methods of euthanasia on ChE activity. Water temperature (17.2, 20.9, and 24.6°C), stress, long-term storage (up to 180 days), postmortem changes, and method of euthanasia had no effect on ChE activity of walleye. There was a strong positive correlation (r = 0.87) between whole body ChE activity and total length (7.2–17.9 mm) for larval walleye, but a negative correlation between brain ChE activity and total length (59–164 mm) for juvenile walleye (r = 0.75). Because size, age, and development may affect ChE activity, fish of similar size should be used when evaluating the effects of ChE inhibitors. If fish of similar size are not available, it is recommended that relations between size, age, and development be understood so estimates of variation in ChE activity can be made. Received: 25 June 2001/Accepted: 19 January 2002  相似文献   

10.
Two hundred sediment samples were collected and their toxicity evaluated to aquatic species in a previous study in the agriculturally dominated Central Valley of California, United States. Pyrethroid insecticides were the main contributors to the observed toxicity. However, mortality in approximately one third of the toxic samples could not be explained solely by the presence of pyrethroids in the matrices. Hundreds of pesticides are currently used in the Central Valley of California, but only a few dozen are analyzed in standard environmental monitoring. A significant amount of unexplained sediment toxicity may be due to pesticides that are in widespread use that but have not been routinely monitored in the environment, and even if some of them were, the concentrations harmful to aquatic organisms are unknown. In this study, toxicity thresholds for nine sediment-associated pesticides including abamectin, diazinon, dicofol, fenpropathrin, indoxacarb, methyl parathion, oxyfluorfen, propargite, and pyraclostrobin were established for two aquatic species, the midge Chironomus dilutus and the amphipod Hyalella azteca. For midges, the median lethal concentration (LC50) of the pesticides ranged from 0.18 to 964 μg/g organic carbon (OC), with abamectin being the most toxic and propargite being the least toxic pesticide. A sublethal growth endpoint using average individual ash-free dry mass was also measured for the midges. The no–observable effect concentration values for growth ranged from 0.10 to 633 μg/g OC for the nine pesticides. For the amphipods, fenpropathrin was the most toxic, with an LC50 of 1–2 μg/g OC. Abamectin, diazinon, and methyl parathion were all moderately toxic (LC50s 2.8–26 μg/g OC). Dicofol, indoxacarb, oxyfluorfen, propargite, and pyraclostrobin were all relatively nontoxic, with LC50s greater than the highest concentrations tested. The toxicity information collected in the present study will be helpful in decreasing the frequency of unexplained sediment toxicity in agricultural waterways.  相似文献   

11.
Accelerated eutrophication is common to many freshwater and marine environments and often co-occurs with the presence of anthropogenic chemicals. However, the toxic effects of common chemical stressors such as herbicides in the presence of elevated nutrients are not well understood for most aquatic flora, particularly vascular species. To provide insight, field-collected Vallisneria americana Michx. (wild celery) were sequentially exposed to three nutrient concentrations for 3 months and then to nominal 11 and 110 μg L−1 atrazine for 96 h. Nutrient concentrations (combined NH4 +, NO2 , NO3 , PO4 ) were based on ambient concentrations in the St. Johns River (FL) and ranged from 0.013 to 0.668 mg L−1. Nutrient pretreatment potentiated the toxicity of atrazine as determined by chlorophyll fluorescence activity. Electron transport rates (ETR) were significantly less (48–59%) for plants pretreated with low and ambient nutrient levels in the presence of an average of 107.5–128.1 μg L−1 atrazine. Significant ETR reductions were also observed for plants exposed to an average of 11.4 μg L−1 atrazine after exposure to nutrients three times the ambient concentration in the St. Johns River. The results indicate the importance of considering the presence of nutrients in chemical hazard assessments, particularly for phytotoxicants and nontarget vascular plants.  相似文献   

12.
Laboratory toxicity data contrasting responses of aquatic organisms to insecticides are important for focusing on sensitive species (steepest exposure-response slope) exposed to aqueous concentrations of these insecticides in field studies. These data also allow prediction of expected responses of aquatic species to a range of insecticide concentrations in situ. Aqueous 48-h toxicity tests were performed to contrast responses of Daphnia magna Straus, Hyalella azteca Saussure, Chironomus tentans Fabricius, and Pimephales promelas Rafinesque to acetylcholinesterase-inhibiting insecticides: chlorpyrifos, aldicarb, and chlordane. As expected, invertebrates tested (H. azteca, C. tentans, and D. magna) were ≥ 200 times more sensitive than the vertebrate P. promelas to chlorpyrifos exposures. H. azteca was approximately 3.5 times more sensitive to chlorpyrifos (453% mortality/μg/L) than D. magna (128% mortality/μg/L). For both aldicarb and chlordane, C. tentans was the most sensitive species tested (2.44 and 2.54% mortality/μg/L, respectively). Differences in chlordane potency for test species varied only by a factor of approximately 2–3 (0.88% mortality/μg/L for H. azteca to 2.54% mortality/μg/L for C. tentans). Although point estimates of population responses such as LC50s, NOECs, and LOECs are of some utility for predicting effects of pesticides in aquatic systems, exposure-response slopes are also useful for extrapolation of laboratory data to diverse field situations, especially where sediment sorption may regulate insecticide exposure or bioavailability. Received: 15 May 1997/Accepted: 9 September 1997  相似文献   

13.
Acute toxicity tests with Cd, Pb, Hg, and methyl parathion were developed to compare the sensitivity of the rotifer Euchlanis dilatata with other model organisms used in aquatic ecotoxicology. Cd was the most toxic chemical (LC50 = 14.8 μg L−1), while methyl parathion was the least toxic (LC50 = 864.2 μg L−1). E. dilatata was more sensitive that other rotifer species, particularly of the genera Brachionus and Lecane. However, E. dilatata was less sensitive to mercury and methyl parathion than the crustacean, Daphnia magna. The high sensitivity of E. dilatata suggests that it may be an adequate benthic model to use in toxicity assessments of metal-contaminated sediments.  相似文献   

14.

Decabromodiphenyl ether (BDE-209), a member of a major group of brominated flame retardants, is detected in aquatic environments at considerable levels and induces physiological and toxic effects on aquatic plants. In this study, the physiological responses induced by and the toxic effects of BDE-209 at different concentrations (0, 0.2, 0.5 and 1.0 mg L−1) in Lythrum salicaria were examined. OJIP transient curves indicated that BDE-209 treatment negatively affected photosystem II (PSII) grouping. Additionally, the results showed that BDE-209 inhibited seedling development and elevated reactive oxygen species (ROS), phosphorylated histone H2AX (γ-H2AX), malondialdehyde (MDA) levels and antioxidative enzyme activities in the roots and shoots of L. salicaria. The results revealed that BDE-209 exposure contributed to ROS accumulation, which was considered as the probable toxicity mechanism. The current results provided an insight into the development of L. salicaria with high BDE-209 tolerance.

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15.
Impact of Atrazine on Chlorpyrifos Toxicity in Four Aquatic Vertebrates   总被引:2,自引:0,他引:2  
Atrazine has been shown previously to potentiate chlorpyrifos toxicity in selected invertebrates. This study examined interactions of atrazine and chlorpyrifos in four aquatic vertebrates. Organisms were exposed to binary mixtures of atrazine and chlorpyrifos during toxicity bioassays. Inhibition of cholinesterase (ChE) enzyme activity and chlorpyrifos uptake kinetics were also examined with and without atrazine exposure. Atrazine alone did not affect organisms at concentrations up to 5000 μg/L; however, the presence of atrazine at 1000 μg/L did result in a significant increase in the acute toxicity of chlorpyrifos in Xenopus laevis. Mixed results were encountered with Pimephales promelas; some bioassays showed greater than additive toxicity, while others showed an additive response. No effect of atrazine on chlorpyrifos toxicity was observed for Lepomis macrochirus and Rana clamitans. Atrazine did not affect ChE activity or chlorpyrifos uptake rates, indicating that these toxicodynamic and toxicokinetic parameters may not be related to the mechanism of atrazine potentiation of chlorpyrifos toxicity. Based on the results of this study, it does not appear that a mixture toxicity of atrazine and chlorpyrifos at environmentally relevant concentrations presents a risk to the vertebrate organisms examined in this study.  相似文献   

16.
Margosan-O, an insecticide formulated from extracts of neem tree (Azadirachta indica) seed kernels, besides being toxic, also has feeding, oviposition-deterring, and growth-inhibitory effects on insects. This product, registered in the United States for ornamental plants, has been proposed for food crop use. However, little information exists on its effects on aquatic organisms. This study investigated toxicity of Margosan-O to the mosquito Culex spp., a possible target species, and to nontarget species—two crustaceans, Daphnia magna, Hyalella azteca, and a dipteran, Chironomus riparius. The 48-h EC50 value of 105 mg L−1 for Culex spp. was significantly more toxic than for C. riparius (281 mg L−1), not significantly different from D. magna (125 mg L−1) but was significantly less toxic than for H. azteca (71 mg L−1). A concentration of 20–30 mg L−1 caused growth inhibitory effects in Culex spp. and C. riparius larvae and 40 and 84 mg L−1 affected growth and reproduction in H. azteca and D. magna, respectively. Margosan-O may not be suitable for mosquito control since the concentrations required to control emergence may have some nontarget effects. Alternatively, the agricultural application of Margosan-O is also not expected to reduce the survival or produce growth and reproductive effects in nontarget aquatic organisms. However, based on estimated concentrations of less than 10 mg L−1 in adjacent shallow bodies of water and recommendations for repeated applications, there should be concern that the threshold for chronic toxicity is too narrow. Received: 9 April 1997/Accepted: 7 April 1998  相似文献   

17.
The frequent use of pesticides in agricultural and commercial settings has led some researchers to devote their attention to studying the effects of mixtures of these compounds as they co-occur in the environment. Recent studies have demonstrated the potentiating effects of triazine herbicides, such as atrazine and its analogs, to the toxicity of a variety of organophosphate (OP) insecticides. One such OP insecticide, chlorpyrifos, has been the topic of much concern because of its prevalence in the environment. This study focused on examining the effects of 10 select triazine herbicides at concentrations of 1 μmole/L (approximately 200 μg/L) to chlorpyrifos with Hyalella azteca. The compounds selected include atrazine, three of its degradation products, and six other herbicide active ingredients. Toxicity tests were performed using a two-way analysis of variance matrix design with effect levels determined by way of probit analysis. Atrazine was found to have the greatest acutely lethal effect to H. azteca, followed by its closest degradation product, deethylatrazine. Two of the six atrazine analogs, simazine and cyanazine, also showed significant effects to the insecticide’s toxicity. Synergistic ratios (SRs) were calculated to compare the effect magnitudes for each of the herbicides. The highest ratio obtained was with atrazine (SR = 1.42). A majority of the past studies involving mixtures of triazines and OPs have examined the potentiation effects of active-use triazine herbicides on Chironomus species. However, compared with the acute effects previously obtained for Chironomus species, H. azteca show a higher tolerance to the presence of the triazine herbicides, even at levels often considered as being at the high end of environmentally relevant concentrations. When coupled with past studies from our laboratory, this research helps to provide a better understanding of the toxic effects of herbicide–insecticide interactions.  相似文献   

18.
Current-use pesticides (CUPs) and banned organochlorine compounds (OCCs) were measured in precipitation (snowpack and rain) and lake sediments from two national parks in the Western United States to determine their occurrence and distribution in high-elevation environments. CUPs frequently detected in snow were endosulfan, dacthal, and chlorothalonil in concentrations ranging from 0.07 to 2.4 ng/L. Of the OCCs, chlordane, hexachlorobenzene, and two polychlorinated biphenyl congeners were detected in only one snow sample each. Pesticides most frequently detected in rain were atrazine, carbaryl, and dacthal in concentrations from 3.0 to 95 ng/L. Estimated annual deposition rates in one of the parks were 8.4 μg/m2 for atrazine, 9.9 μg/m2 for carbaryl, and 2.6 μg/m2 for dacthal, of which >85% occurred during summer. p,p’-DDE and p,p’-DDD were the most frequently detected OCCs in surface sediments from lakes. However, concentrations were low (0.12 to 4.7 μg/kg) and below levels at which harmful effects for benthic organisms are likely to be observed. DDD and DDE concentrations in an age-dated sediment core suggest that atmospheric deposition of DDT and its degradates, and possibly other banned OCCs, to high-elevation areas have been decreasing since the 1970s. Dacthal and endosulfan sulfate were present in low concentrations (0.11 to 1.2 μg/kg) and were the only CUPs detected in surface sediments. Both pesticides were frequently detected in snow, confirming that some CUPs entering high-elevation aquatic environments through atmospheric deposition are accumulating in lake sediments and potentially in aquatic biota as well.  相似文献   

19.
Endocrine disrupting chemicals (EDCs) are introduced into the aquatic environment through industrial and municipal effluents along with urban and agricultural runoffs. Exposure of aquatic organisms to EDCs may lead to hormonal disruption and adverse health effects. The goals of our study were: to collect anchovy and mussel samples from the coastal region of Karachi, to use the yeast estrogen screen (YES) bioassay in estimating xeno-estrogen content in these samples, and to investigate if the bioassay could be used to quantify known amounts of 17β-estradiol (E2) injected into cod and salmon fillets. Results of the studies showed that mussel estrogenic activity in Karachi decreased in the order of Buleji point 1 (8.91 ± 4.77, mean ± SD) > Paradise point 1 (1.72 ± 0.81) > Paradise point 2 (0.61 ± 0.84) ng E2 equivalents/g wet wt (p < 0.05). By comparison, anchovy estrogenic activity at Korangi/Phitti Creek was much higher than at Manora. Together, these results confirmed previous reports that both Buleji point 1 and Korangi/Phitti Creek were the most contaminated areas of Karachi. The YES bioassay was only a semi-quantitative method in determining the contents of xeno-estrogens in aquatic organisms; it consistently overestimated the amounts of E2 injected into cod and salmon fillets due to additive and/or non-additive interactions between E2 and endogenous estrogens. Nevertheless, the YES bioassay was able to identify the contaminated sites in the coastal region of Karachi.  相似文献   

20.
The responses of algae to herbicides depend on the sensitivity of each species, but competition within algal communities may be an important regulator of the effects of herbicides on aquatic systems. The impact of herbicides on algae also depends on abiotic factors like nutrients, light, and temperature. We examined the tolerance of the alga Oscillatoria limnetica Lemmermann to the photosystem II inhibitor herbicide atrazine under different culture conditions to assess those interactions between herbicides and biotic and abiotic factors. The density of the cyanobacterium O. limnetica was determined in natural phytoplanktonic assemblages and in unialgal cultures in medium containing 10 μg/L atrazine. Experiments (total of 13) were conducted in spring and early and late summer, during which the effect of atrazine varied in nature and intensity. The growth of the cyanobacterium was always inhibited in cold experiments, whereas it was stimulated in warm experiments within the natural phytoplankton assemblage, but unaffected in pure culture. Laboratory experiments with unialgal culture showed that the sensitivity to atrazine increased as the temperature decreased. Phytoplankton community structure, interactions between species, and environmental parameters (e.g., temperature) are important factors controlling the responses of cyanobacteria to the herbicide. These interactions between sensitivity to herbicides and environmental factors may reduce or emphasize the effects of pollution in aquatic systems. Thus, the ecotoxicological relevance of herbicides in aquatic systems is quite complex and cannot be assessed by single-species short-term laboratory toxicological tests. Received: 29 January 1999/Accepted: 7 June 1999  相似文献   

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