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21.
Irena Šutiaková Natália Kovalkovičová Jaroslav Legáth Václav Šutiak 《Journal of environmental science and health. Part. B》2013,48(7):606-611
The fungicide tolylfluanid (N - dichlorofluoromethylthio-N′, N - dimethyl - N - p - tolylsulfamide), was investigated by cytokinesis-block micronucleus assay. Tolylfluanid at the lowest concentration (1 × 10? 6mol L? 1)did not influence significantly the frequency of micronuclei in sheep lymphocyte cultures in comparison with control (32.33 ± 3.51/1000 binucleated cells versus 30.33 ± 2.82/1000 binucleated cells in dimethylsulfoxide (DMSO) control, P = 0.44). Higher tolylfluanid concentrations (1 × 10? 4 and, 1 × 10? 5 mol L? 1) resulted in a significant dose-dependent increase in the number of micronuclei in comparison with control (74.00 ± 13.00/1000 binucleated cells and 52.67 ± 10.12/1000 binucleated cells versus 30.33 ± 2. 82/1000 binucleated cells in DMSO control, P = 0.005 and 0.02, respectively, ANOVA followed by Tukey test P < 0.05). Many of the treated cells also possessed multiple micronuclei. Tolylfluanid did not affect the nuclear division index at all treatment concentrations. Our in vitro results thus demonstrate that tolylfluanid had a significant genotoxic effect at only the highest concentration tested. 相似文献
22.
Jianshe Tang Min Zhang Guohua Cheng Ang Li Yitong Lu 《Journal of environmental science and health. Part. B》2013,48(8):707-712
Diethyl (carboxymethyl) phosphonate (DECP) was used as the hapten to develop an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) for detecting organophosphorus pesticides (OPs). Conjugator of DECP with bovin serum albumin (BSA) was used as the immunogen for producing the polyclonal antibodies (PcAbs). Three antisera were obtained after the immune procedure. Characterization studies of the PcAbs indicated that the titer of antiserum-1 was highest in 3 antisera, and antiserum-1 had high affinity and specificity to the parathion, dichlorvos and pirimiphos. The IC-ELISA showed an IC50 of 0.428 μ g/mL with a detection limit of 0.0125 μ g/mL to parathion. The assay also indicated that the IC50 values of pirimiphos and dichlorvos were 0.331 μ g/mL and 1.25 μ g/mL respectively, and the detection limits of pirimiphos and dichlorvos were 0.0116 μ g/mL and 0.048 μ g/mL respectively. Recoveries of parathion, pirimiphos and dichlorvos spiked into water samples ranged from 90% to 160%. The results indicated that the ELISA could be a convenient and supplemental analytical tool for monitoring OPs residues in environmental water samples. 相似文献
23.
K.M.S. Sundaram A. Sundaram L. Sloane 《Journal of environmental science and health. Part. B》2013,48(6):1341-1362
Abstract The effect of two tracer dyes [Erio Acid Red (EAR) and Acid Black 48 (AB‐48)] on initial deposits and persistence of Bacillus thuringiensis subsp. kurstaki (Btk) toxin (delta‐endotoxin) was studied after spraying two commercial formulations, Foray® 48B and Foray® 76B, over potted white spruce [Picea glauca (Moench) Voss] seedlings, at a dosage rate of 30 billion international units (BIU) per ha. Spray was applied using a spinning disc atomizer calibrated to deliver droplet sizes similar to those utilized in ultra‐low‐volume (ULV) treatments in operational insect control programs. The sprayed seedlings were left outdoors at the Sault Ste. Marie laboratory for 18 days under natural conditions of sunlight, wind and rainfall. Initial deposits and persistence of delta‐endotoxin protein in spruce foliage were determined by immunoassay [enzyme linked immunosorbent assay (ELISA)] quantification of the delta‐endotoxin. The total protein (inactive plus active) and delta‐endotoxin (active protein) concentrations in the two formulations were determined by a gravimetric procedure and by ELISA respectively. The initial deposit levels of the toxin on foliage were not markedly affected by the addition of either of the two tracer dyes, and showed only a narrow range of 1521 to 1625 ng/g foliage (fresh weight) for Foray 48B, and 1789 to 2056 ng/g for Foray 76B. However, the persistence of the toxin was significantly influenced by the presence of the dyes. The toxin persisted in foliage only for 7 d post‐spray When the EAR dye was added to Foray 48B, compared to 10 d when no dye was added. The average half‐life (DT50) of disappearance was 17.4 h for Foray 48B with EAR, and 20.9 h when no dye was present. In contrast, the situation was reversed in Foray 76B, since the duration of persistence was 10 d when EAR was added to Foray 76B, compared to 7 d when no dye was added. The average DT50 was 27.9 h for Foray 76B with EAR, and 22.2 h without the dye. Persistence was the longest (14 d) when the AB‐48 dye was added to Foray 76B, and the DT50 was 44.9 h. 相似文献
24.
Robert Kase Peter D. Hansen Birgit Fischer Werner Manz Peter Heininger Georg Reifferscheid 《Environmental science and pollution research international》2009,16(1):54-64
Background, aim, and scope The enzyme-linked receptor assay (ELRA) detects estrogenic and anti-estrogenic effects at the molecular level of receptor
binding and is a useful tool for the integrative assessment of ecotoxicological potentials caused by hormonally active agents
(HAA) and endocrine disrupting compounds (EDC). The main advantage of the ELRA is its high sample throughput and its robustness
against cytotoxicity and microbial contamination. After a methodological adaptation to salinity of the ELRA, according to
the first part of this study, which increased its salinity tolerance and sensitivity for 17-β-estradiol, the optimised ELRA
was used to investigate 13 native sediments characterised by different levels of salinity and chemical contamination. The
applicability of the ELRA for routine analysis in environmental assessment was evaluated. Salinity is often a critical factor
for bioassays in ecotoxicological sediment assessment. Therefore, salinity of the samples was additionally adjusted to different
levels to characterise its influence on elution and binding processes of receptor-binding substances.
Materials and methods The ELRA was carried out with the human estrogen receptor α (ER) in a 96-well microplate format using the experimental setup
known from the competitive immunoassay based on ligand–protein interaction. It is an important improvement that a physiologically
relevant receptor was used as a linking protein instead of an antibody. The microplates were coated with a 17-β-estradiol-BSA
conjugate, and dilution series of estradiol and of native sediment samples were added and incubated with the ER. After a washing
step, a biotinylated mouse anti-ER antibody was added to each well. Receptor binding to estradiol, agonistic and antagonistic
receptor binding, were determined by a streptavidin-POD-biotin complex with subsequent measurement of the peroxidase activity
at the wavelength of 450 nm using a commercial ELISA multiplate reader. The sediment elutriates and pore water samples of
sediments were tested in a dilution series to evaluate at which dilution step the receptor-binding potential ends. In the
elution process (see Section 2.1 to 2.2), a method was developed to adjust the salinity to the levels of the reference testings, which offers an appropriate option
to adjust the salinity in both directions. Statistical evaluation was made with a combination of the Mann–Whitney U test and the pT-method.
Results This part of the study characterised the environmental factor ‘salinity’ for prospective applications of the ELRA. Using reference
substances such as 17-β-estradiol, the ELRA showed sigmoid concentration-effect relations over a broad range from 0.05 μg/l
to 100 μg/l under physiological conditions. After methodological optimisation, both sensitivity and tolerance of the assay
against salinity could be significantly raised, and the ELRA became applicable under salinity conditions up to concentrations
of 20.5‰. The mean relative inter-test error (n = 3) was around 11% with reference substances and below 5% for single sediments elutriates in three replicates each. For
sediment testings, the pore water and different salinity-adjusted elutriates of 13 sediments were used. A clear differentiation
of the receptor-binding potential could be reached by application of the pT-method. Thereby, pT-values from one to six could
be assigned to the sediments, and the deviation caused by the different salinity conditions was one pT-value. The mean standard
deviation in the salinity adaptation procedure of the elutriates was below 5%.
Discussion Although the ELRA has already been used for assessments of wastewater, sludge and soil, its applicability for samples to different
salinity levels has not been investigated so far. Even if the ELRA is not as sensitive as the E-screen or the YES-assay, with
regard to reference substances like 17-β-estradiol, it is a very useful tool for pre-screening, because it is able to integrate
both estrogenic as well as anti-estrogenic receptor-binding effects. According to the results of sediment testing, and given
the integrative power to detect different directions of effects, the ELRA shows sufficient sensitivity and salinity tolerance
to discriminate receptor-binding potentials in environmental samples.
Conclusions The optimised ELRA assay is a fast, cost-effective, reliable and highly reproducible tool that can be used for high-throughput
screening in a microplate format in detecting both estrogenic and anti-estrogenic effects. Additionally, the ELRA is robust
against microbial contaminations, and is not susceptible towards cytotoxic interferences like the common cell-culture methods.
The general applicability and sufficient sensitivity of the ELRA was shown in freshwater environments. Marine and brackish
samples can be measured up to salinity levels of 20.5‰.
Recommendations and perspectives In view of the proven sensitivity, functionality and the fastness of the ELRA, it is recommendable to standardise the test
method. At the moment, no adequate in vitro test procedure exists which is standardised to DIN or ISO levels. The E-screen
and the yeast estrogen/androgen screens (YES/YAS) sometimes underlie strong cytotoxic effects, as reported in the first part
of this study. Further development of an ELRA assay using human androgen receptors appears to be very promising to gain information
about androgenic and anti-androgenic effects, too. This would offer a possibility to use the ELRA as a fast and reliable pre-screening
tool for the detection of endocrine potentials, thus minimising time and cost-expensive animal experiments. 相似文献
25.
26.
The present work deals with the application of genotoxicity biomarkers by means of the Comet assay in haemocytes and spermatozoa of the crustacean Gammarus elvirae exposed in vivo to heavy metals. Furthermore, a basal levels (BLs) study of DNA damage in the two cell types considered for two different gammarids species, G. elvirae and Echinogammarus veneris, was carried out. It is important to identify factors that influence the outcome of the assay in order to obtain reliable and reproducible results usable for risk assessment purposes. Our results highlight that the Italian legal limits for Hg and Pb, respectively, 0.5 and 50?µg/L, are inadequate for establishing safety thresholds in the aquatic environment. Furthermore, the freshwater invertebrate G. elvirae, used for the first time to measure the effect of genotoxicants, is a good candidate for evaluating the genotoxicity damage induced by heavy metals. Our results concerning spermatozoa show excessively variable responses and high BLs. 相似文献
27.
28.
基于微板藻毒性试验测定5个有机磷农药与4个三嗪类农药的单个及联合毒性.根据半数效应浓度(EC50),对斜生栅藻96h生长抑制的毒性大小顺序为:西草净>阿特拉津>扑灭通>苯嗪草酮>草甘膦>敌敌畏>磷胺>乙酰甲胺磷>甲胺磷.这表明直接干扰光合作用电子传输的三嗪除草剂的藻毒性明显大于有机磷农药.以通用浓度加和作为参考模型,三嗪类农药按EC50和EC10(10%效应浓度)浓度比的混合物对斜生栅藻呈现加和毒性.有机磷农药按EC50和EC10浓度比的混合物在低浓度呈现加和毒性,在高浓度呈现协同毒性.有机磷与三嗪类农药按EC50和EC10浓度比的混合物在低浓度为加和毒性,在高浓度为协同毒性. 相似文献
29.
利用彗星实验检测渤海区主要入海河流遗传毒性.以虾虎鱼为受试生物,暂养在河口水样中,染毒48h,取外周血细胞,运用彗星实验检测外周血细胞内DNA损伤程度,以尾相(TM)作为DNA损伤程度指标,并据此评估入海河流邻近海域遗传毒性风险.实验结果表明,入海河流中的特征污染物可导致虾虎鱼外周血细胞的DNA损伤,且损伤程度可以通过彗星实验定量分析,同时该试验方法操作简便、快速、灵敏度高,能够反映出多种污染因子的综合致毒能力.因此,通过彗星实验建立实验室检测入海河流遗传毒性方法具有可行行和创新性. 相似文献
30.
Ulrich Harréus MD PhD Philipp Baumeister Norbert Kleinsasser Maximilian Reiter Beatrice Bachmeier Christoph Matthias 《毒物与环境化学》2013,95(2):205-214
The etiology of salivary gland malignancies still remains unclear. Metal compounds are of special interest since they show ubiquitous presence in the environment, are present in many working places, and are accepted (co-)carcinogens in some other malignancies. Metals enter the body as xenobiotics by inhalation or ingestion. This study investigated the genotoxic potential of sodium dichromate (Na2Cr2O7), nickel sulfate (NiSO4), cadmium sulfate (CdSO4) and zinc chloride (ZnCl2) on human salivary gland cells and lymphocytes. Macroscopically healthy tissue of salivary glands was harvested from 46 patients during surgery and isolated to single cells by enzymatic digestion. The cells were incubated with Na2Cr2O7, NiSO4, CdSO4 or ZnCl2. Na2Cr2O7 was also incubated in combination with the other metal compounds listed. Carcinogenic and co-carcinogenic effects of cadmium were tested by incubation with Na2Cr2O7 and consecutive repair intervals. DNA damage and repair were evaluated by the Comet assay, determining DNA-strand breaks. The extent of damage was quantified using a digital analysis system. Na2Cr2O7 produced significantly enhanced DNA-strand breaks in human salivary gland tissue and lymphocytes. All other metal compounds exerted no damaging effect on both cell types. Co-incubation of Na2Cr2O7 with the other metals revealed a significant additive effect only for CdSO4. Specific analysis of the influence of cadmium showed a reduction of DNA-repair after Na2Cr2O7-induced strand breaks in salivary gland cells. This study provides evidence that exposure to distinct metals may significantly contribute to malignant salivary gland tumors. In consequence, further studies as epidemiological and toxicological data are warranted to determine the role of distinct metals as potential (co-) carcinogens. 相似文献