Luminescent Bacteria Toxicity Research Articles

Potential and mechanism of disinfection by-products removal in drinking water by bubbling corona discharge

Disinfection by-products (DBPs) with significant teratogenic and carcinogenic properties have become a growing concern among the public. As an efficient and environmentally friendly technology, non-thermal plasma offers potential for removing emerging micro-pollutants. In this study, the degradation performance of bubbling corona discharge was evaluated on 24 halogenated alicyclic and aliphatic DBPs present in drinking water at concentrations ranging from ng/L to μg/L. The degradation of DBPs followed pseudo-first-order kinetics with rate constants (kobs) in the descending order of halonitromethanes (HNMs), halogenated benzoquinones (HBQs), haloacetonitriles, trihalomethanes (THMs), haloketones, halogenated aldehydes, and haloacetic acids (HAAs). THMs, HNMs, and HBQs were effectively removed within 5 min under a discharge power of 28 W. Degradation rates achieved by plasma treatment surpass those of other conventional treatment technologies. The required energy consumption was in the range of 5–30 kW·h/m3/order.Furthermore, the study investigated the effects of discharge power, initial concentration, and economic analysis on the degradation of four selected DBPs as representatives of mono-, di- and multi-carbon-containing DBPs, namely chloroform (TCM) and bromoform (TBM), tribromoacetic acid (TBAA), and 2,3,5,6-tetrachloro-1,4-benzoquinone (TetraC-BQ). Reactive radicals in the plasma system were investigated using electron paramagnetic resonance, optical emission spectroscopy, fluorimetry, and radical scavengers. Hydrated electrons and hydroxyl radicals played an important role in the removal of DBPs. The intermediates generated during the degradation of TCM, TBM, TBAA, and TetraC-BQ were identified, and the possible degradation pathways for mono- and binary C-DBPs and HBQs were deduced. The breakdown of HBQs did not produce secondary contamination with aliphatic DBPs. The carbon in DBPs was primarily converted to formic acid, acetic acid, and oxalic acid, and the halogens were mainly converted to halogen ions. Additionally, luminescent bacteria toxicity testing confirmed that plasma treatment could reduce the acute toxicity of water samples. These findings demonstrate the potential of plasma treatment as a post-treatment device at the household level.

Shift of combined ecotoxicity index in petroleum polluted soils during a bacterial remediation

Introduction: Bioremediation has been shown to be an effective strategy for removing toxic pollutants from the environment, particularly organic chemicals such as petroleum hydrocarbons. This paper investigates the changes in toxicity of petroleum-contaminated soil as a result of microbial remediation processes.Methods: Changes in the ecotoxicity of the contaminated soil were examined using a plant, earthworm, enzyme activity and luminescent bacteria toxicity tests.Results: The results showed that bioremediation could effectively degrade petroleum hydrocarbon (C10–C40) pollutants. After 42 days of remediation, the petroleum hydrocarbon (C10–C40) content of Group A (bioaugmented polluted wetland soil) decreased from 1.66 g/kg to 1.00 g/kg, and the degradation rate was 40.6%. The petroleum hydrocarbon (C10–C40) content of Group B (bioaugmented polluted farmland soil decreased from 4.00 g/kg to 1.94 g/kg, and the degradation rate was 51.6%. During the microbial remediation progress, the ecological toxicity of petroleum-contaminated soil first increased and then decreased. The photosynthetic pigment content index in the higher plant toxicity test, the earthworm survival index and the soil catalase activity all showed good agreement with the relative luminescence index of extracted DCM/DMSO in the luminescent bacterial toxicity test. The soil toxicity decreased significantly after remediation. Specifically, the photosynthetic pigment content of wheat were inhibited in the soil during the whole process (remediation for 42 days), and decreased to the minimum on remediation day 21. The 7-day and 14-day survival rate of earthworms in Group A and Group B gradually decreased in the soil remediation process, and then gradually increased, survival rate at the end of remediation was higher than at the beginning. Soil catalase activity was significantly negatively correlated with petroleum hydrocarbon (C10–C40) content (−0.988, −0.989). The ecological toxicity of contaminated soil reached to the maximum on the 21st day of remediation, relative luminosity of luminescent bacteria in dichloromethane/dimethyl sulfoxide extracts from Group A and Group B were 26.3% and 16.3%, respectively.Conclusion: Bioremediation could effectively degrade petroleum hydrocarbon (C10–C40) pollutants. Wheat photosynthetic pigment content, earthworm survival rate, soil catalase activity and relative luminescence of luminescent bacteria can better indicate the ecological toxicity of petroleum-contaminated soil in bioremediation process.

Evolvement of carbamazepine under solar irradiation in the presence of chlorine: Efficiency, influences, degradation pathways and ecotoxicity

Carbamazepine (CBZ) and residual chlorine coexist in wastewater discharged from wastewater treatment plants (WWTPs), while the mechanism of their photochemical transformation in the natural aquatic environment is still unclear. In this study, an approach combining experiment and quantum chemical calculation was used to explore the degradation mechanism of CBZ under chlorine/solar irradiation. The degradation of CBZ was not evident during disinfection alone or sunlight irradiation. However, CBZ was rapidly degraded under chlorine/solar irradiation (kobs = 0.463 min−1 at pH = 8.0). Quenching and probe experiments showed that the efficient removal of CBZ under chlorine/solar irradiation was credited with the production of ·Cl, ·OH, and O3, in which ·Cl (61.95%) and ·OH (33.14%) contributed significantly. The reaction mechanism of CBZ degradation was postulated, in conjunction with the identification of degradation products by HPLC/MS analysis and density functional theory. Among them, the primary degradation way was acridine production under the action of ·Cl and ·OH, and the decarboxylation process was the rate-limiting step. In addition, bioassay results indicated that the luminescent bacteria toxicity and genotoxicity of CBZ after chlorine/solar irradiation increased. Based on the identified transformation products and the quantitative structure-activity relationship, the elevated ecotoxicity was attributed to the TP209 and TP179 generated by pathway 1, and then controlling the decarboxylation process by adjusting pH and temperature to reduce the high toxic degradation products were proposed for ecological safety.

The impact of natural sunlight irradiation on the biotoxicities of different molecular sizes EfOM/SRNOM and its relationship with spectral and molecular level parameters

Natural sunlight irradiation is regarded as an efficient and low-carbon method for controlling the biotoxicity of effluents from domestic wastewater treatment plants (WWTPs). Dissolved organic matter in WWTPs effluent (EfOM) is responsible for the non-specific biotoxicity of effluent. In the present study, the variation in spectral characteristics, molecular composition, luminescent bacteria toxicity, and genotoxicity of EfOM of different molecular sizes (MOSs) during natural sunlight irradiation were investigated from a systematic perspective, and the standard natural organic matter from the Suwannee River (SRNOM) was synchronously assessed for comparative purposes. To further explore the cause of the biotoxicity changes, the relationships between the spectral or molecular level parameters (obtained from FT-ICR MS analysis) and biotoxicity were assessed using correlation analysis. The molecules in <1 kDa EfOM with lower molecular weight, higher unsaturation degree, and higher humification and fluorescence had higher luminescent bacteria toxicity under sunlight irradiation. However, in the <1 kDa SRNOM, the molecules which were characterized by higher humification and fluorescence had higher luminescent bacteria toxicity. The notable genotoxicity reduction of EfOM under sunlight irradiation was attributed to the photochemical degradation of components with a high unsaturation degree. Such findings could enable ecological safety improvement of aquatic environments using natural sunlight.

Ferrate (VI) Oxidation Is an Effective and Safe Way to Degrade Residual Colistin - a Last Resort Antibiotic - in Wastewater.

The rise of novel mcr mobile resistance genes seriously threatens the use of colistin as a last resort antibiotic for treatment of multidrug-resistant Gram-negative bacterial infections in humans. Large quantities of colistin are released annually into the environment through animal feces. This leads to environmental toxicity and promotes horizontal transmission of the mcr gene in aqueous environments. We examined colistin degradation catalyzed by the presence of strong oxidant Fe (VI). We found almost complete colistin degradation (>95%) by Fe (VI) at initial colistin levels of 30 μM at a molar ratio of Fe (VI): colistin of 30 using an initial pH 7.0 at 25°C for 60 min. The presence of humic acid did not alter the degradation rate and had no significant impact on the removal of colistin by Fe (VI). Quantitative microbiological assays of Fe (VI)-treated colistin solutions using Escherichia coli, Staphylococcus aureus, and Bacillus subtilis indicated that the residual antibacterial activity was effectively eliminated by Fe (VI) oxidation. Luminescent bacteria toxicity tests using Vibrio fischeri indicated that both colistin and its degradation products in water were of low toxicity and the products showed decreased toxicity compared to the parent drug. Therefore, Fe (VI) oxidation is a highly effective and environment-friendly strategy to degrade colistin in water.

Modelling of possible tanker accident oil spills in the Istanbul Strait in order to demonstrate the dispersion and toxic effects of oil pollution.

Countries located on the Black Sea coast perform most of their sea trade through the Istanbul Strait (IS). Approximately 50,000 ships pass through the IS each year, with crude oil tankers making up the majority. Thus, the aim of the study is to determine the acute toxic effect of oil pollution that may occur as a result of crude oil tanker accidents in the IS. By utilising data related to accidents that have occurred in the IS, locations of concentrated tanker accidents, or "hot spots," were determined by Kernel Density Analysis. Subsequently, the distribution of potential leaks following an oil tanker accident, within these hot spots, is modelled with GNOME software. Finally, acute toxicity caused by oil pollution in the marine ecosystem is determined by Aliivibrio fischeri luminescent bacteria toxicity test. In this research, 5 hot spots are identified, where the maximum calculated amount of oil that can reach the coastline after 72h is 3096 metric tons. Similarly, oil pollution can affect a total coastline of 30-35km. Furthermore, it was determined that after the oil was diluted in seawater, at a ratio of 1:200,000, the toxic effects decrease (EC50 above 100mg/L), yet the chronic effects may still continue. The results of this study may serve as a reference for coastal state authorities to develop emergency response plans. Having this valuable knowledge of where high-risk accidents are most concentrated, where the accidents occur intensely, which areas can be affected by the pollution, the duration of the pollution effects, and the distance between the areas, will help determine the number of intervention stations to be installed, their locations, and equipment to be installed to the stations.

Micropollutants and biological effects as control indexes for the operation and design of shallow open-water unit ponds to polish domestic effluent

Additional control indexes should be considered for the operation and design of post-treatment systems, as the wastewater treatment objectives are developing toward protecting the safety of ecological environments. In this study, two control indexes were selected and examined systematically in pilot-scale shallow open-water unit (SOWU) ponds for domestic effluent polishing: micropollutants and biotoxicities. The total risk quotient (RQTotal ≤ 1) and effect-based trigger value (EBT) were set as the thresholds for known micropollutants and biological effects, respectively. The results showed that RQTotal of micropollutants (n = 46) could be mitigated to an acceptable level and the luminescent bacteria toxicity was in compliance with the EBT after SOWU polishing in the warm season. The reduction of micropollutants and biotoxicities in the SOWUs both fit the k–C* model well (R2 > 0.9) in the warm and cold seasons. Finally, the k–C* model integrated with the control indexes was developed to design the SOWU dimensions, and the results indicated that a pond area of 21.7–108.5 m2 was required for every 1 m3/d of effluent when micropollutants were set as the control index, while a pond area of 3.6–18.2 m2 was required when luminescent bacteria toxicity was set as the control index.

Bioremediation of metal-contaminated soils by microbially-induced carbonate precipitation and its effects on ecotoxicity and long-term stability

Bioremediation using microorganisms is a promising technique to remediate soil contaminated with heavy metals. In this study, Sporosarcina pasteurii (S. pasteurii) bioremediation by mixing method was used to remediate soils contaminated with lead (Pb), zinc (Zn) and cadmium (Cd). A significant reduction of heavy metal leaching concentrations was observed in S. pasteurii bio-treated samples. Furthermore, urease hydrolyzing bacteria have additional advantages of accelerating metal precipitation by increasing pH. The soluble-exchangeable Pb, Zn and Cd was reduced by 33.3 % ∼ 85.9 %, 21.4 % ∼ 66.0 %, 13.6 % ∼ 29.9 % respectively after bioremediation. The primary objective of metal stabilization was achieved by reducing the bioavailability through immobilizing the Pb, Zn and Cd in the urease-driven carbonate precipitation. Luminescent bacteria toxicity experiments revealed that the metal toxicity of contaminated soil was reduced after bioremediation using S. pasteurii. When subjected to severe environmental conditions, S. pasteurii bioremediation was superior to chemical precipitation technology in terms of long-term stability.

Kinetics and Mechanism of Sucralose Degradation in Water Using UV-activated Persulfate Process

The degradation of emerging pollutant artificial sweetener sucralose (SUC) using UV/persulfate (UV/PS). The effects of several process parameters, including UV light intensity, PS dosage, pH, and anion concentration, were also investigated. The degradation products and their toxicity during the UV/PS process were further analyzed and evaluated. It is reported that, compared with single UV or PS, the degradation of SUC by UV/PS was more obvious. The degradation rate constants increased with an increase in the light intensity and PS dosage. The SUC degradation could be improved under neutral conditions. The background ions NO3- and HCO3- could inhibit the degradation process, while Cl- and SO42- ions could accelerate the process. Sixteen intermediate products were identified using high-resolution mass spectrometry (HRMS) and GC-MS. Hydroxylation, oxidation, ether cracking, and other reactions were involved. A degradation path was further proposed. Moreover, luminescent bacteria toxicity test and ECOSAR prediction showed that the intermediates with higher toxicity could be produced during UV/PS, which could pose a potential threat to the ecological environment.

Iron-nitrogen co-doped carbon nanotubes decorated with Cu2O possess enhanced electronic properties for effective peroxymonosulfate activation

Exploiting the full potential of copper-based nanoparticles in the activation of peroxymonopersulfate (PMS) is a great challenge due to their insufficient dispersity and electronic properties. We report here a novel iron‑nitrogen co-doped carbon nanotube (FNC) modified with a Cu2O nanocomposite (Cu2O/FNC) that exhibits ultrahigh catalytic performance in the activation of PMS to degrade fluconazole (~95%). Catalytic performance evaluation illustrated that Cu2O/FNC also has wide pH applicability (3.0–11.0), long-term stability and excellent adaptability. In addition, luminescent bacteria toxicity tests confirm that Cu2O/FNC/PMS significantly reduced the acute biotoxicity of various recalcitrant pollutants (reduced by 45–83%). By identifying the reactive oxygen species (ROS) and catalytic performance for various pollutants, we propose that pollutants that interact weekly with activators are mostly destroyed by sulfate radicals and hydroxyl radicals, whilst both radical and non-radical routes were involved in the degradation of pollutants that were easily adsorbed. By modifying Cu2O with FNC, several crucial properties such as the specific surface area, surface defects, active sites and the charge transfer rate were significantly improved, leading to excellent catalytic performance for pollutant removal. Finally, a reasonable reaction mechanism is advanced for the fluconazole degradation pathway. This study not only develops a novel PMS oxidation system for fluconazole degradation, but also provides a new strategy to improve the reactivity and applicability of PMS activators by combining radical and non-radical activation pathways.

Removal of trace organic pollutants (pharmaceuticals and pesticides) and reduction of biological effects from secondary effluent by typical granular activated carbon

Residual trace organic pollutants (TOPs) and associated biological effects from secondary effluent (SE) are attracting much attention because of their safety concerns. Granular activated carbon (GAC) adsorption, due to its low cost and high efficiency, is widely applied for further wastewater treatment, but its selective removals of TOPs and biological effects are poorly understood. In the present study, the surface physicochemical characteristics of four types of typical GACs were investigated, and their correlation with luminescent bacteria toxicity was discussed. Based on the biological effect control, shell GAC, with a great adsorption capacity and high functional group contents was selected for further study, including for the removal of fluorescent dissolved organic matter (DOM), 21 TOPs, and 3 biological effects. The shell GAC showed a promising property of removing fluorescent DOM and TOPs. The total concentration of 21 detected TOPs, including 12 pesticides and 9 pharmaceuticals, achieved 82% removal when 30 g/L shell GACs was added. Individual chemicals removal by GAC adsorption was not well described by an individual parameter (e.g., logD, molecular size, charge, functional groups), but rather by a variety of physical and chemical interactions among TOPs, DOM, and GAC. The biological effects from SE were mainly caused by TOPs and DOM. Hence, shell GACs also showed high removal efficiencies of luminescent bacteria toxicity, genotoxicity, and photosynthetic inhibition effect. The removal mechanisms of the three biological effects from SE were deeply discussed. Therefore, the GAC treatment is considered to be one of the most suitable options to ensure the ecological safety of discharged wastewater, because it can effectively control DOM, TOPs, and associated biological effects.

Biological Toxicity Effects of Soil Pollution Caused by Galvanized Wastewater Based on Vibrio Qinghaiensis sp.-Q67.

Objective To establish a rapid diagnosis method for the biological toxicity of soil, accurately and rapidly evaluate the toxicity of contaminated sites and identify the dominant pollutants. Methods Take the soil pollution of a galvanized factory as an example, while the metal concentration level was analyzed and detected, a rapid biological toxicity detection method based on the acute toxicity test of luminescent bacteria （Vibrio qinghaiensis sp.-Q67） was established, and the dominant pollutants were identified by stepwise multiple regression. Results The pollutants came from wastewater and metal plating fragments directly discharged from the manufacturing line of the factory. The concentration of those pollutants was correlated with the acute toxicity of Vibrio qinghaiensis sp.-Q67. The dominant pollutants in the study were zinc （Zn）, aluminum （Al） and copper （Cu）. Conclusion The luminescent bacteria toxicity test method based on Vibrio qinghaiensis sp.-Q67 can conveniently and rapidly assess the degree of toxic damage of polluted soil and identify the dominant pollutants and can be applied to the acute toxicity evaluation of polluted soil.

Evaluation of ecotoxicological effects of benzophenone UV filters: Luminescent bacteria toxicity, genotoxicity and hormonal activity

The widespread use of organic ultraviolet (UV) filters in personal care products raises concerns about their potentially hazardous effects on human and ecosystem health. In this study, the toxicities of four commonly used benzophenones (BPs) UV filters including benzophenone (BP), 2-Hydroxybenzophenone (2HB), 2-Hydroxy-4-methoxybenzophenone (BP3), and 2-Hydroxy-4-methoxybenzophenone-5-sulfonicacid (BP4) in water were assayed in vitro using Vibrio fischeri, SOS/umu assay, and yeast estrogen screen (YES) assay, as well as in vivo using zebrafish larvae. The results showed that the luminescent bacteria toxicity, expressed as logEC50, increased with the lipophilicity (logKow) of BPs UV filters. Especially, since 2HB, BP3 and BP4 had different substituent groups, namely -OH, -OCH3 and -SO3H, respectively, these substituent functional groups had a major contribution to the lipophilicity and acute toxicity of these BPs. Similar tendency was observed for the genotoxicity, expressed as the value of induction ratio=1.5. Moreover, all the target BPs UV filters showed estrogenic activity, but no significant influences of lipophilicity on the estrogenicity were observed, with BP3 having the weakest estrogenic efficiency in vitro. Although BP3 displayed no noticeable adverse effects in any in vitro assays, multiple hormonal activities were observed in zebrafish larvae including estrogenicity, anti-estrogenicity and anti-androgenicity by regulating the expression of target genes. The results indicated potential hazardous effects of BPs UV filters and the importance of the combination of toxicological evaluation methods including in vitro and in vivo assays.

Forms Distribution and Ecotoxicity of Three Forms of Sulfonamides in Root-Soil Interface of Maize

Forms distribution and ecotoxicity of sulfonamides in root-soil interface of maize were studied by chemical analysis and luminescent bacteria toxicity assay. The results indicated that the water soluble residues were the main form in rhizosperic soils, and the organic solvent extractable residues were the main form in far-rhizosperic soils. The three forms of sulfonamides showed relatively lower toxicity to luminescent bacteria with the relative inhibition rates less than 30%. The toxicity of water soluble residues was lower than that of the organic solvent extractable residues, and the bound residues showed the lowest toxicity to luminescent bacteria. The contents and relative inhibition rates of water soluble residues in rhizosperic soils were higher than those in far-rhizosperic soils, while the contents and relative inhibition rates of organic solvent extractable and bound residues in rhizosperic soils were lower than those in far-rhizosperic soils.

Managing Drilling Wastes: Detoxification of Two Formaldehyde-Releasing Biocides

Summary An increased awareness of the environmental impact and operational costs associated with freshwater usage and wastewater disposal in energy production has shifted industry interest toward replacing freshwater sources with lower-quality or recycled water in oilfield applications, and has highlighted the importance of addressing toxicity as part of a successful waste-management plan. Poor-quality and recycled waters often contain high concentrations of bacterial assemblages, which can cause operational challenges such as corrosion, slime formation, and souring. Microbial-control agents, such as biocides, are subsequently necessary to manage bacteriological problems. However, these chemicals are highly reactive and can react indiscriminately with biological targets, making their toxicity both a performance metric and an ecological, human-health, and disposal concern. The luminescent-bacteria toxicity test presented in this work, for instance, is a key regulatory parameter in the pumpoff and landspray disposal of drilling fluids in Alberta, Canada. Considering the necessary toxicity of biocides, controlled detoxification following use is a pertinent factor in responsible hazard management. Formaldehyde-releasing agents are the most widely used category of microbial-control additives that slowly and continuously release small amounts of formaldehyde, a toxic environmental pollutant and known human carcinogen. This research evaluated the acute (short-term) aquatic toxicity of two liquid formaldehyde-releasing biocides, identified necessary parameters for their detoxification, measured the resulting change in their toxicity over time, and used regulatory requirements for toxicity testing set by the Alberta Energy Regulator (AER) for drilling-waste management to evaluate the practical relevance of this detoxification to waste-management practices. The additives investigated were a tetrakis(hydroxymethyl)phosphonium sulfate (THPS)-based product, and a 1,3-dimethylol-5,5-dimethylhydantoin (DMDMH)-based product. Laboratory results suggest that the THPS-based additive was more toxic than the DMDMH-based additive on a percent volume basis, and pH was an important factor in THPS toxicity. Aeration alone decreased the toxicity of DMDMH over the course of the experiment, while a combination of aeration and pH increase were necessary to decrease the toxicity of THPS over the same time period. This work presents a proof of concept for a relatively simple and cost-effective detoxification of the evaluated additives, highlights the key parameters for this process, and uses toxicity-threshold levels referenced by the AER drilling-waste-management directive to evaluate their application in waste-assessment practices.

Toxicity bioassays for water from black-odor rivers in Wenzhou, China

Following urbanization, a large number of urban rivers were contaminated and turned to black-odor rivers. The traditional approach for detecting water quality is based on chemical or physical analysis. However, biological toxicity of black-odor water has been less addressed. As two typical black-odor rivers, Jiushanwai River (JS) and Shanxia River (SX) are tributaries of Wen-Rui Tang River in Wenzhou (south of China). The eco-safety of the urban rivers was evaluated by bioassay for water toxicity in this study. Ten and 5 sampling sites were respectively set along JS and SX. Water samples were collected monthly from October 2010 to October 2011. The general physical and chemical parameters of river water were monitored. In order to investigate the ecotoxicological effects of black-odor water, the following bioassays were used: (1) Fish acute toxicity test (Danio rerio, comprehensive toxicity), (2) luminescent bacteria bioassay (Qinghaiensis vibrio, toxicity to bacteria), and (3) tropical claw embryo assay (Xenopus tropicalis, embryo toxicity). Biotoxicity of black-odor rivers water was demonstrated by D. rerio, Q. vibrio, and X. tropicalis embryos. Toxicological effects of black-odor water were respectively shown by mortality of zebrafish, and by the relative inhibitory light rate of luminescent bacteria. However, luminescent bacteria were more sensitive to inspect biotoxicity than zebrafish. In X. tropicalis embryos test, toxicological effects of black-odor water were mostly shown by embryos' survival rate and teratogenic rate. Bioassay results showed that toxicity of SX water was higher than that of JS water, especially in summer. Statistical analysis of luminescent bacteria toxicity test showed that biotoxicity of SX and JS was high in summer, but low in winter and spring. The seasonal changes of water toxicity of the black-odor river were positively correlative with changes of water temperature (p < 0.05), and related to pH and ammonium nitrogen of water. Typical black-odor river water displays different degrees of biotoxicity to D. rerio, luminescent bacteria, and X. tropicalis embryos. The ecotoxicological risk of black-odor rivers was demonstrated in urban area, which suggests bioassay is necessary for evaluation of water quality. In the present study, spatial and seasonal bioassay for toxicity of JS and SX provides a complete example for evaluation of urban rivers.

Control of the Survival and Activity of Luminescent Bacteria*

Luminescent bacteria toxicity test has attracted considerable attention in the past ten years.However,maintaining the stabilization of the activity of luminescent bacteria is a demanding task,which seriously limits the application of such diagnostics in on-line determination and analysis.In this study,a control method of maintaining the stabilization of the intensities of Photobacterium phosphoreum and P.leiognathi was investigated by adding various cryoprotectants.In this method,the bacteria in the exponential growth phase was centrifuged,re-suspended in the cold skimmed milk solution and stored at 5 ℃.Three cold storage methods were studied and compared,and they were the skimmed milk-bacteria suspension,freeze-dried bacteria suspension made by rehydrating freeze-dried bacteria powder and fresh bacteria solution being stored at 5 ℃.The results indicated that the storage life using skimmed milk bacteria suspension could reach 7 days,with relative luminosity 93%.This method could significantly improve the stabilization of the luminescent bacteria,and it could be applied in on-line toxicity monitor to improve continuous operation time.

Energetic Salts Based on Dipicrylamine and Its Amino Derivative

Energetic salts based on dipicrylamine and its amino derivative were synthesized. All salts were fully characterized by multinuclear NMR spectroscopy ((1)H, (13)C), vibrational spectroscopy (IR), and elemental analysis. Ethylenediammonium di-DPA (DPA=dipicrylamine) and 1,3-diaminoguanidinium DPA were further confirmed by single-crystal X-ray diffraction. These salts exhibit reasonable physical properties, such as high densities (1.71-1.81 g cm(-3)), good thermal stabilities (T(d) =155-285 °C), and low solubilities in water. The impact sensitivity of 1-methyl-3,4,5-triamino-1,2,4-triazolium DPA is lower than that of 2,4,6-trinitrotoluene (TNT), and for some other energetic salts their impact sensitivities are comparable to that of TNT. Based on experimental densities and theoretical calculations carried out by using the Gaussian 03 suite of programs, all the salts have calculated detonation pressures (22.5-27.8 GPa) and velocities (7226-7917 m s(-1)) that exceed those of conventional TNT. The toxicities of these salts measured by luminescent bacteria toxicity tests are much lower than that of TNT, and two binary eutectic mixtures with melting points that fall between 70 and 100 °C were identified.

Whole effluent assessment of industrial wastewater for determination of bat compliance

The applicability of the Whole Effluent Assessment concept for the proof of compliance with the "best available techniques" has been analysed with paper mill wastewater from Germany by considering its persistency (P), potentially bio-accumulative substances (B) and toxicity (T). Twenty wastewater samples from 13 paper mills using different types of cellulose fibres as raw materials have been tested in DIN or ISO standardised bioassays: the algae, daphnia, luminescent bacteria, duckweed (Lemna), fish-egg and umu tests with lowest ineffective dilution (LID) as test result. The potentially bio-accumulative substances (PBS) were determined by solid-phase microextraction and referred to the reference compound 2,3-dimethylnaphthalene. Usually, a primary chemical-physical treatment of the wastewater was followed by a single or multi-stage biological treatment. One indirectly discharged wastewater sample was pre-treated biologically in the Zahn-Wellens test before determining its ecotoxicity. No toxicity or genotoxicity at all was detected in the acute daphnia and fish egg as well as the umu assay. In the luminescent bacteria test, moderate toxicity (up to LIDlb=6) was observed. Wastewater of four paper mills demonstrated elevated or high algae toxicity (up to LIDA=128), which was in line with the results of the Lemna test, which mostly was less sensitive than the algae test (up to LIDDW=8). One indirectly discharged wastewater sample was biodegraded in the Zahn-Wellens test by 96% and was not toxic after this treatment. Low levels of PBS have been detected (median 3.27 mmol L(-1)). The colouration of the wastewater samples in the visible band did not correlate with algae toxicity and thus is not considered as its primary origin. Further analysis with a partial wastewater stream from thermomechanically produced groundwood pulp (TMP) revealed no algae or luminescent bacteria toxicity after pre-treatment of the sample in the Zahn-Wellens test (chemical oxygen demand elimination 85% in 7 days). Thus, the algae toxicity of the respective paper mill cannot be explained with the TMP partial stream; presumably other raw materials such as biocides might be the source of algae toxicity. Comparative data from wastewater surveillance of authorities confirmed the range of ecotoxicity observed in the study. Wastewater from paper mills generally has no or a moderate ecotoxicity (median LID 1 and 2) while the maximum LID values, especially for the algae and daphnia tests, are considerably elevated (LIDA up to 128, LIDD up to 48). Wastewater from paper mills generally is low to moderately ecotoxic to aquatic organisms in acute toxicity tests. Some samples show effects in the chronic algae growth inhibition test which cannot be explained exclusively with colouration of the samples. The origin of elevated algae ecotoxicity could not be determined. In the algae test, often flat dose-response relationships and growth promotion at higher dilution factors have been observed, indicating that several effects are overlapping. At least one bioassay should be included in routine wastewater control of paper mills because the paper manufacturing industry is among the most water consuming. Although the algae test was the most sensitive test, it might not be the most appropriate test because of the complex relationship of colouration and inhibition and the smooth dose-effect relationship or even promotion of algae growth often observed. The Lemna test would be a suitable method which also detects inhibitors of photosynthesis and is not disturbed by wastewater colouration.

Microbial toxicity and impacts on soil enzyme activities of pesticides used in potato cultivation

In the conventional cultivation of potatoes, weed control and the control of potato late blight caused by Phytophthora infestans are carried out by the application of herbicides and fungicides. We investigated the impacts of the herbicides metribuzin and linuron and the fungicide fluazinam on soil microbiota in microcosms, in mesocosms and in the field. The toxicity of each pesticide in solution was assessed using the luminescent bacteria test and in soil by a solid phase modification. In microcosm tests, the microbial activity and biomass were estimated by measuring several soil enzyme activities together with soil ATP content. In the mesocosm tests, the separate addition of each pesticide and the simultaneous use of all the pesticides were investigated. We monitored the impacts on ten different soil enzyme activities and measured soil toxicity with the luminescent bacteria test separately in the 5 cm top layer and in the layer from 5 to about 20 cm below the surface. During one season, the impact of the use of pesticides was monitored in the field in plots receiving pesticides for the third consecutive year and in control plots cultivated without the use of pesticides for the 3 preceding years. The pesticide concentrations were monitored in each experiment. The luminescent bacteria toxicity test revealed a strong inhibition by fluazinam. In microcosms the herbicides increased several enzyme activities but metribuzin inhibited luminescence in the bacterial test. Fluazinam was highly toxic in microcosms. In the mesocosm with combined use of pesticides, decreased activities of some enzymes were observed first in the surface soil and at harvesting also deeper in the soil. In the mesocosm experiment with separate use of pesticides, less impacts were observed. In the field experiment the pesticides decreased seven enzyme activities, when calculated per soil fresh weight but activities of four enzyme decreases if calculated per soil organic matter. Controlling weeds by herbicides decreased weed growth and the decreases in enzyme activities were interpreted to be due to the lack of the stimulating impact of weed roots. A strong inhibition in the soil toxicity test and continuing bioavailability of fluazinam were detected throughout the experiments even after winter in the field.