Comparison of Three Bacterial Toxicity Assays for Imidazolium-Derived Ionic Liquids

Ionic liquids have become leading candidates for replacing many common organic solvents used in the chemical process industry. There is, however, a general lack of toxicology data relevant to wastewater treatment facility microbes for these compounds. In this study, we performed three bacterial-based toxicity assays on several imidazolium-derived compounds as well as the precursor compound 1-methylimidazole. Two of the assays, the Shk1 and Microtox assays, are used as surrogate assays for toxicity to bacterial respiration in activated sludge wastewater treatment plants. The third assay was a direct measure of the effect of toxicity on mixed bacterial culture respiration, using a commercially available consortium of naturally occurring bacteria to obtain IC50 values for direct comparison to the EC50 values from the surrogate assays. The Shk1 assay is based on a genetically engineered bioluminescent Pseudomonas bacterium and is more highly correlated with the respiration inhibition than the Microtox assay. The Shk1 assay gave EC50 values more similar to IC50 values from the bacterial respiration inhibition assay for the compounds tested in this work. The Shk1 EC50 values were similar to that of 1-butanol, an alcohol with an alkyl chain length similar to that of the cation of the tested compounds, which were 1-butyl-3-methylimidazolium bromide ([bmim][Br]), 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]), 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, ([bmim][Tf2N]), and the precursor compound 1-methylimidazole, and were generally smaller than those typical of aromatic organic solvents.     

Comparative Study of Two Bioassays for Applications in Influent Wastewater Toxicity Monitoring

Bioluminescent bacteria-based assays can be used for influent wastewater toxicity monitoring for biological wastewater treatment systems. The most thoroughly studied bioluminescent bacteria-based test is the Microtox? assay. However, the response to toxicants of Photobacterium phosphoreum, the marine bacterial strain used in this assay, is different from that of the activated sludge microorganisms. We developed a continuous influent wastewater monitoring system based on the bioluminescent bacterium Shk1, a genetically modified Pseudomonad isolated from the activated sludge in an industrial wastewater treatment plant. The Shk1 toxicity data were correlated with the Microtox? toxicity data for 79 organic compounds and the two toxicity assays were compared. The Shk1 assay is less sensitive than the Microtox? assay and could therefore be more suitable for influent wastewater toxicity monitoring.     

Comparison of Bacterial Bioluminescence with Activated Sludge Oxygen Uptake Rates during Zinc Toxic Shock Loads in a Wastewater Treatment System

The use of a bioengineered bioluminescent bacterium (Shk1) for monitoring zinc toxicity was evaluated with samples from a municipal activated sludge wastewater treatment plant and in a bench-scale activated sludge system. Bioluminescent measurements were compared with oxygen uptake rates of activated sludge samples. In batch experiments with activated sludge, the Zn EC50 for Shk1 bioluminescence was 16 mg/L, while the Zn EC50 for activated sludge OURs was approximately 58 mg/L. In the bench-scale system, the influent Zn concentrations tested were 50 and 200 mg/L in toxic shock loads of about 4 h duration. Soluble Zn transport through the influent, aeration basin, and clarifier was able to be monitored by the decrease in Shk1 bioluminescence. However, bioluminescence in samples from the aeration basin decreased faster than activated sludge specific oxygen uptake rates. Differences in responses of Shk1 and the activated sludge community may be due to differences in the assay conditions, the growth forms, physiology of the organisms, or previous cultivation conditions.     

Toxicity Estimation of Phenolic Compounds by Bioluminescent Bacterium

Phenolic compounds are environmentally important due to their extensive use in various industries, presence in wastewaters, and potential toxicity. When phenolic compounds reach the activated sludge processes of publicly owned treatment works, they can cause upsets in the operations of the treatment plants. A continuous toxicity testing system was developed based on the bioluminescent bacterium Shk1. The toxicities of 23 phenolic compounds to Shk1 were studied and the 5 min EC50 values were obtained. Quantitative structure-activity relationship models based on the logarithm of the octanol-water partitioning coefficient [log(Kow)] were established for estimating the toxicity of phenolic compounds and the model validity was verified.     

Recurrence of alcoholic liver disease after liver transplantation

The acute toxicity of six metals [Hg, Cd, Cu, Zn, Cr(II), and Cr(VI)] to Daphnia magna neonates was evaluated using three different test media (Elendt M7, a complex medium containing EDTA; ASTM hard water and EEC, two simple media free of chelators). The EC50 values, at both 24 and 48 h, obtained for Zn, Cr(II), and Cr(VI) were similar in all of the media tested. Hg was more toxic in ASTM than in M7 and in EEC media. The toxicity of Cd and Cu was similar in ASTM and EEC media and higher when evaluated in M7 medium. Thus, M7 should be used only carefully for the toxicity evaluation of mixtures and effluents containing metals. It is recommended, however, that it be excluded from use in tests evaluating samples of unknown composition, or those known to contain Cu and Cd. For the metals tested in this study, a factor of five applied to each 48-h EC50 would be sufficient in order to attain the respective acute NOECs for immobility.     

市政污泥及其热处理渣对CHO-K1细胞的毒性研究

评估市政污泥资源化产物的健康风险对其安全规范利用具有指导意义。分别对某污水处理厂的污泥进行了热解和好氧发酵处理,运用中国仓鼠卵巢细胞（CHO-K1）对市政污泥及其热处理渣开展细胞毒性试验和遗传毒性试验,评估其对典型哺乳动物细胞的危害和潜在风险。结果显示,与原污泥样品相比,污泥热解渣和好氧发酵渣的细胞毒性和遗传毒性均降低。与原污泥样品相比,RTCA细胞增殖试验结果显示,污泥热解渣和好氧发酵渣对CHO-K1细胞的生长抑制性明显减弱;CCK-8细胞毒性试验结果显示,污泥热解渣和好氧发酵渣的细胞相对存活率分别增加了161%和137%;Caspase 3细胞凋亡蛋白酶活力试验结果显示,暴露于污泥热解渣和好氧发酵渣中的CHO-K1细胞的Caspase 3凋亡蛋白酶的活性分别降低了29.3%和20.1%;彗星试验结果显示,污泥热解渣和好氧发酵渣的尾长分别缩短了64.5%和25.9%,尾矩分别减少了78.4%和28.1%。研究结果表明,热处理技术（热解处理和好氧发酵处理）可减少市政污泥的细胞毒性和遗传毒性,降低市政污泥对生态环境和人体健康的潜在风险。     

Differentiation of Allium carlaviruses isolated from different parts of the world based on the viral coat protein sequence

Common primers which amplify the 3' terminal genomic RNAs of Allium carlaviruses were designed based on the nucleotide sequence of shallot latent virus (SLV), garlic latent virus (GLV) and garlic common latent virus (GCLV). A total of fifteen cDNAs encoding the coat protein (CP) of the carlaviruses, including the biologically identified isolates SLV, GLV and GCLV as well as viruses from infected Allium plants cultivated in different parts of the world, were amplified by RT-PCR with the common primers. The cDNAs were then cloned and sequenced. The predicted viral CP amino acid sequence as well as the nucleotide sequence revealed that SLV and GLV, previously considered as separate viruses on the basis of their biological and physical properties, belong to the same species of the genus Carlavirus. Both viruses are clearly differentiated from GCLV. In addition, every SLV and GLV isolate from the Allium plants in Taiwan showed characteristic and common variations in their CP sequences, suggesting the possible presence of geographical variants. However, no apparent sequence variations of SLV and GLV related to their host plant species, including A. sativum, A. wakegi, A. chinense, A. fistulosum, A. cepa and A. ampeloprasum, were observed. These findings suggested that the sequence variations observed in the respective virus isolates do not correlate with the specificity of their infectivities for Allium species.     

An experimental model for the rational treatment of arrhythmias: a clinical study with quinidine

Fourteen patients with supraventricular and/or ventricular ectopic beats selected by clinical, dynamic ECG and exercise test, underwent a basal continuous ECG recording (Holter) and then were given 400 mg of quinidine orally. The concentrations of the drug were determined in blood samples taken 30 minutes before, and 1, 2, 4, 6, 8, 12, and 24 hours after administration. On the same day patients were also monitored by continuous ECG recording (Holter). One week later, after a washout period, the same subjects were treated with 200 mg of quinidine orally 4 times a day for two weeks. On day 1, 2, 4, 7 and 14 quinidine was determined in plasma, and on day 7 and 14 a 24 hour ECG recording was also done. On the basis of pharmacokinetic parameters obtained from the acute test, the chronic levels of the drug were predicted. Predicted values were superimposable to those observed. Thus, the kinetics of single-dose quinidine is able to predict the steady-state levels after repeated dosing. During acute administration there was an increase in QTc interval and a decrease in ectopic beats. These effects correlated with a sigmoid pattern with acute quinidine levels. Concentrations producing 50% of the effect (EC50) could be calculated from these curves. In spite of similar steady-state blood levels of quinidine, some patients after chronic therapy did not respond to treatment (non responders). Non responders could be predicted by the acute test because they had greater EC50 values of QTc increase: patients with EC50 greater than 2 mg/L were all non responders.     

Bricks from Petroleum Effluent Treatment Plant Sludge: Properties and Environmental Characteristics

A voluminous and hazardous sludge containing a high amount of hydrocarbons and several trace metals is generated in petroleum oil field effluent treatment plants. The aim of this study was to utilize the sludge in preparing environmentally acceptable masonry bricks in a commercial brick plant. The effect of the sludge on the plasticity behavior of the brick raw mix was investigated. The addition of the sludge reduces the requirement of process water and fuel. The fired bricks meet all the requirements of the Indian Standard Specification. The bricks were subjected to toxicity characteristics leaching protocol leaching tests. Most of the toxic metals are fixed in the vitrification process and the leachates values meet the Environmental Protection Agency’s requirement for recycling of hazardous materials.     

Neural plasticity, neuropeptides and anxiety in animals--implications for understanding and treating affective disorder following traumatic stress in humans

Phenolic composition, toxicity and biodegradability of three different phenolic leachates/samples was studied. Samples A and C were the leachates from the oil-shale industry spent shale dumps at Kohtla-J?rve, Estonia. Sample B was a laboratory-prepared synthetic mixture of 7 phenolic compounds mimmicking the phenolic composition of the leachate A. Toxicity of these 3 samples was analyzed using two photobacterial test (BioTox and Microtox), Daphnia test (DAPHTOXKIT F pulex) and rotifiers' test (ROTOXKIT F). All the LC50 values were in the range of 1-10%, leachate A being the most toxic. The growth and detoxifying potential (toxicity of the growth medium was measured using photobacterial tests) of 3 different phenol-utilizing bacteria and acclimated activated sludges was studied in shake-flask cultures. 30% leachate A (altogether 0.6 mM total phenolic compounds) was too toxic to rhodococci and they did not grow. Cell number of Kurthia sp. and Pseudomonas sp. in 30% leachate A increased by 2 orders of magnitude but despite of the growth of bacteria the toxicity of the leachate did not decrease even by 7 weeks of cultivation. However, if the activated sludge was used instead of pure bacterial cultures the toxicity of the 30% leachate A was eliminated already after 3 days of incubation. 30% samples B and C were detoxified by activated sludge even more rapidly, within 2 days. As the biodegradable part of samples A and B should be identical, the detoxification of leachate A compared to that of sample B was most probably inhibited by inorganic (e.g. sulphuric) compounds present in the leachate A. Also, the presence of toxic recalcitrant organic compounds in the leachate A (missed by chemical analysis) that were not readily biodegradable even by activated sludge consortium should not be excluded.     

Orientation of interphase chromosomes as detected by Giemsa C-bands

The orientation of Giemsa C-bands has been studied in mitotic and interphase cells of Allium cepa. A sativum and of Aloe vera. The C-bands in these three species are located at the telomeres, secondary constriction region of the nucleolar chromosomes and the centromeric regions, respectively. Observations in A. cepa and Aloe indicate clearly that the interphase chromosomes are non-random in their orientation and possibly maintain their telophase configuration through the attachment of telomeres and perhaps of kinetochores with the nuclear membrane. Electron micrographs of onion cells also reveal that certain heterochromatic segments are associated with the nuclear membrane.--The nucleolar interstitial C-bands in A. sativum remain free in the nucleoplasm and may come close to each other due to heterochromatic attraction. Such a heterochromatic attraction is also evident between telomeric regions and between centromeres. However, a two by two attachment could not be noticed. A diagrammatic representation of the orientation of interphase chromosomes has been presented.     

Toxicity of N-substituted aromatics to acetoclastic methanogenic activity in granular sludge

N-substituted aromatics are important priority pollutants entering the environment primarily through anthropogenic activities associated with the industrial production of dyes, explosives, pesticides, and pharmaceuticals. Anaerobic treatment of wastewaters discharged by these industries could potentially be problematical as a result of the high toxicity of N-substituted aromatics. The objective of this study was to examine the structure-toxicity relationships of N-substituted aromatic compounds to acetoclastic methanogenic bacteria. The toxicity was assayed in serum flasks by measuring methane production in granular sludge. Unacclimated cultures were used to minimize the biotransformation of the toxic organic chemicals during the test. The nature and the degree of the aromatic substitution were observed to have a profound effect on the toxicity of the test compound. Nitroaromatic compounds were, on the average, over 500-fold more toxic than their corresponding aromatic amines. Considering the facile reduction of nitro groups by anaerobic microorganisms, a dramatic detoxification of nitroaromatics towards methanogens can be expected to occur during anaerobic wastewater treatment. While the toxicity exerted by the N-substituted aromatic compounds was closely correlated with compound apolarity (log P), it was observed that at any given log P, N-substituted phenols had a toxicity that was 2 orders of magnitude higher than that of chlorophenols and alkylphenols. This indicates that toxicity due to the chemical reactivity of nitroaromatics is much more important than partitioning effects in bacterial membranes.     

Biological and phytochemical evaluation of plants. XIV. Antiinflammatory evaluation of 163 species of plants

One hundred and seventy-seven plant extracts, representing 163 species of plants and/or fungi, were evaluated in rats to determine their antiinflammatory activity using the carrageenin-induced pedal edema assay. Of the 163 species of plants and/or fungi tested, 17 exhibited between 30--39 per cent inhibition of inflammation, 21 between 40-49 per cent, 15 between 50--59 per cent, four between 60--69 per cent, and two gave greater than 70 per cent inhibition of carrageenin-induced pedal edema.     

In vitro genotoxic effects of the insecticide deltamethrin in human peripheral blood leukocytes: DNA damage ('comet' assay) in relation to the induction of sister-chromatid exchanges and micronuclei

Deltamethrin, a synthetic dibromo-pyrethroid insecticide, is extensively used in agriculture, forestry and in household products because of its high activity against a broad spectrum of insect pests (both adults and larvae), its low animal toxicity and its lack of persistence in the environment. Data on the genotoxicity and carcinogenicity of deltamethrin are rather controversial, depending on the genetic system or the assay used. The aim of this study was to further evaluate the potential genotoxic activity of deltamethrin. The in vitro genotoxicity of deltamethrin has been evaluated by assessing the ability of the insecticide to damage DNA (as evaluated using the single-cell microgel-electrophoresis or 'comet' assay) or induce sister-chromatid exchanges (SCE) and micronuclei (MN) in human peripheral blood leukocytes. All treatments were conducted with and without the presence of an external bioactivation source (+/- S9mix). The results indicate that deltamethrin, in the presence of metabolic activation (+ S9mix), is able to induce DNA damage (double- and single-strand breaks, alkali-labile sites and open excision repair sites) as revealed by the increasing tail moment values observed with increasing doses. The frequency of SCE and MN were not statistically increased in deltamethrin-treated cells as compared to controls, both with and without S9mix. However, lower deltamethrin doses were tested, as compared to 'comet' assay, because of cytotoxicity.     

Evaluation of toxicity indicators in rat primary astrocytes, C6 glioma and human 1321N1 astrocytoma cells: can gliotoxicity be distinguished from cytotoxicity?

A comparison was made of rat primary astrocytes, C6 glioma cells pre-treated with dibutyryl cyclic AMP, and the human astrocyte 132N1 cell line using a range of 40 compounds and the neutral red (NR) assay. The 40 chemicals included substances known to be toxic to astrocytes or neurons, to be generally cytotoxic or not thought to be toxic to nervous tissue. For those compounds which were toxic, changes in glial fibrillary acidic protein (GFAP) levels were measured in the primary and C6 cultures, and changes in vimentin and S-100 measured in the C6 cells. The number of compounds with EC50 values < 2000 microg/ml for the NR assay for the different cell cultures were as follows: primary astrocytes, 19; C6 cells, 15; and 1321N1 cells, 11. The log of the EC50 values for the NR assay for the test compounds between the three cell types was not significantly different at the 5% level by paired Student's t-test. For the toxic substances the correlation coefficients of the EC50 values between primary cells and the C6 or 1321N1 cells were r > 0.5, and between the C6 and 1321N1 cells r > 0.9. For GFAP there was a similar degree of correlation in EC50 values between the different cell types. The GFAP, vimentin and S-100 levels showed similar EC50 values for the toxicants, but were not as sensitive as the NR assay. The toxic substances caused altered morphology in the primary, C6 and 1321N1 cells, with increased branching of cell processes. The combined astrocyte systems identified 8 out of 9 substances reported to be toxic to astrocytes in vivo, together with substances which have general cytotoxic properties. A number of substances (including the 1 out of 9 reported gliotoxic substances), which may primarily affect neurons, which may affect nervous tissue after long-term exposure, or which are not thought to be toxic to nervous tissue, were not detected. The astrocyte systems positively identify gliotoxic and cytotoxic substances and will allow detailed mechanistic studies to be made on the different underlying mechanisms.     

Environmental effects on cellular photosensitization: correlation of phototoxicity mechanism with transient absorption spectroscopy measurements

The presence of actin in eukaryotic nuclei and chromosomes, and especially in higher plant nuclei and chromosomes, has not been well established. We detected actin in meristematic cells of Allium cepa with indirect immunofluorescence technique and observed bright fluorescence in the intact nuclei and chromosomes, indicating that actin is present in the nuclei and chromosomes of the higher plant. We labeled sections of the meristematic cells of A. cepa with immunogold technique, gold particles were found over the whole nuclei and a number of gold particles were concentrated in condensed chromatin and nucleoli, confirming the results of the immunofluoresence observations. We treated the nuclei and chromosomes of A. cepa with DNase I and 2M NaCl and obtained DNA- and histone-depleted nuclei and chromosomes. Indirect immunofluorescence tests showed that the DNA- and histone-depleted nuclei and chromosomes reacted positively with the anti-actin antibodies. These results demonstrate that actin exists not only in intact nuclei and chromosomes, but also in DNA- and histone-depleted nuclei and chromosomes of the plant. In addition, our immuno-fluorescence tests indicate that tropomyosin is present in the nuclei and chromosomes of A. cepa.     

低浓度有机废水活性污泥处理实验技巧的研究

文章针对活性污泥法处理工业废水实验中废水的保质、性能预判等关键的步骤,在实验数据分散条件下,采用平均值为基础来进行定性数据分析。     

Lidocaine toxicity in primary afferent neurons from the rat

Evidence from both clinical studies and animal models suggests that the local anesthetic, lidocaine, is neurotoxic. However, the mechanism of lidocaine-induced toxicity is unknown. To test the hypothesis that toxicity results from a direct action of lidocaine on sensory neurons we performed in vitro histological, electrophysiological and fluorometrical experiments on isolated dorsal root ganglion (DRG) neurons from the adult rat. We observed lidocaine-induced neuronal death after a 4-min exposure of DRG neurons to lidocaine concentrations as low as 30 mM. Consistent with an excitotoxic mechanism of neurotoxicity, lidocaine depolarized DRG neurons at concentrations that induced cell death (EC50 = 14 mM). This depolarization occurred even though voltage-gated sodium currents and action potentials were blocked effectively at much lower concentrations. (EC50 values for lidocaine-induced block of tetrodotoxin-sensitive and -resistant voltage-gated sodium currents were 41 and 101 microM, respectively.) At concentrations similar to those that induced neurotoxicity and depolarization, lidocaine also induced an increase in the concentration of intracellular Ca++ ions ([Ca++]i; EC50 = 21 mM) via Ca++ influx through the plasma membrane as well as release of Ca++ from intracellular stores. Finally, lidocaine-induced neurotoxicity was attenuated significantly when lidocaine was applied in the presence of nominally Ca(++)-free bath solution to DRG neurons preloaded with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Our results indicate: 1) that lidocaine is neurotoxic to sensory neurons; 2) that toxicity results from a direct action on sensory neurons; and 3) that a lidocaine-induced increase in intracellular Ca++ is a mechanism of lidocaine-induced neuronal toxicity.     

Common genetic determinants of halothane and isoflurane potencies in Caenorhabditis elegans

BACKGROUND: Genetics provides a way to evaluate anesthetic action simultaneously at the molecular and behavioral levels. Results from strains that differ in anesthetic sensitivity have been mixed in their support of unitary theories of anesthesia. Here the authors use the previously demonstrated large variation of halothane sensitivities in Caenorhabditis elegans recombinant inbred strains to assess the similarities of the determinants of halothane action with those of another volatile anesthetic, isoflurane. METHODS: The recombinant inbred strains, constructed from two evolutionarily distinct C. elegans lineages, were phenotyped. A coordination assay on agar quantified the sensitivity to the volatile anesthetics; median effective concentrations (EC50s) were calculated by nonlinear regression of concentration-response data and were correlated between the drugs for those strains tested in common. Genetic loci were identified by statistical association between EC50s and chromosomal markers. RESULTS: The recombinant inbred strains varied dramatically in sensitivity to halothane and isoflurane, with a 10-fold range in EC50s. Heritability estimates for each drug were imprecise but altogether high (49-80%). Halothane and isoflurane EC50s were significantly correlated (r=0.71, P < 10(-9)). Genetic loci controlling sensitivity were found for both volatile anesthetics; the most significant determinant colocalized on chromosome V. A smaller recombinant inbred strain study of ethanol-induced immobility segregated different genetic effects that did not correlate with sensitivity to either halothane or isoflurane. CONCLUSIONS: The genetic determinants driving the large variation in anesthetic sensitivity in these C. elegans recombinant inbred strains are very similar for halothane and isoflurane sensitivity.