Comparison of Joint Effect of Acute and Chronic Toxicity for Combined Assessment of Heavy Metals on Photobacterium sp.NAA-MIE.

Predicting the crucial effect of single metal pollutants against the aquatic ecosystem has been highly debatable for decades. However, dealing with complex metal mixtures management in toxicological studies creates a challenge, as heavy metals may evoke greater toxicity on interactions with other constituents rather than individually low acting concentrations. Moreover, the toxicity mechanisms are different between short term and long term exposure of the metal toxicant. In this study, acute and chronic toxicity based on luminescence inhibition assay using newly isolated Photobacterium sp.NAA-MIE as the indicator are presented. Photobacterium sp.NAA-MIE was exposed to the mixture at a predetermined ratio of 1:1. TU (Toxicity Unit) and MTI (Mixture Toxic Index) approach presented the mixture toxicity of Hg2+ + Ag+, Hg2+ + Cu2+, Ag+ + Cu2+, Hg2+ + Ag+ + Cu2+, and Cd2+ + Cu2+ showed antagonistic effect over acute and chronic test. Binary mixture of Cu2+ + Zn2+ was observed to show additive effect at acute test and antagonistic effect at chronic test while mixture of Ni2+ + Zn2+ showing antagonistic effect during acute test and synergistic effect during chronic test. Thus, the strain is suitable and their use as bioassay to predict the risk assessment of heavy metal under acute toxicity without abandoning the advantage of chronic toxicity extrapolation.

Rapid assessment of heavy metal toxicity using bioluminescent bacteria Photobacterium leiognathi strain GoMGm1.

Several commercial test kits such as Microtox, LUMIStox, ToxAlert, Aboatox, and ToxScreen have been widely used for toxicity screening. Though this time saving assays offer excellent sensitivity, cost-effectiveness, and accuracy, these commercial assays are limited in terms of real-time monitoring in Indian coastal environment due to warmer temperatures. This necessitates the need to develop a rapid and accurate assay that can be effectively employed for real time monitoring with respect to heavy metals in the Indian coastal waters. With this objective, the present study was conducted by isolating an indigenous luminescent bacterium from the light organs of chordates Gazza minuta which showed higher luminescence in a wide range of temperatures. The isolate could grow well in the temperature of 30 ± 2 °C and withstand temperature up to 35 ± 2 °C. The isolated bacterium was identified as Photobacterium leiognathi GoMGm1 based on 16S rDNA and luxA gene sequences. The suitable growing medium was optimized using central composite rotational design (CCRD) method to obtain optimal growth and luminescence. The optimized medium exemplified the maximal growth and luminescence of P. leiognathi at OD600 nm of 5.78 ± 0.12 and RLU of 12.49 ± 0.43. The isolate was used to assess the toxicity of several heavy metals. The IC50 values of 0.0051, 1.13, 1.37, 3.1, and 6.68 mg L-1 were observed for the Hg, Cr, Cu, Ni, and Zn, respectively, after 15 min of exposure. Results obtained from principal component analysis (PCA) displayed the present assay's compatibility with other luminescent bacterial assay and commercial Microtox™ assay. Thus, it would the right candidate as an early detection system for heavy metals in aquatic bodies in tropical countries. Schematic representation of the present study.

Acute toxicity assessment of indoor dust extracts by luminescent bacteria assays with Photobacterium Phosphoreum T3.

In the last decades, there has been an increasing concern about the human exposure to indoor dust. Therefore, it is imperative to assess the toxicity of indoor dust and associated dust extracts. In this study, the acute toxicity assessment of indoor dust was performed using a bioluminescence test, with Photobacterium phosphoreum T3 (PPT3) chosen as the test bacterium. The different indoor dust samples were collected from residences, offices, dormitories and laboratories in Shanghai, China. Our data reveal that PPT3 is more active to water-soluble ions and organic contaminants at low concentrations, while extract solutions elicit increased bacterial toxicity at high concentrations. The results of a bioluminescence assay by PPT3 indicated that the dust organic extracts exhibited increased toxicity compared with the water exacts. Dust extracts from the laboratory exhibited the greatest bacterial toxicity when compared with office, dormitory and residence samples. Moreover, office dust exhibited higher bacterial toxicity than residence dust. Furthermore, the comprehensive toxicity of dust on PPT3 was assessed by extracts toxicity -addition (i.e. IRaddition). The calculated values were close to the corresponding experimental data. The bioluminescence test showed the indoor dust samples are weakly toxic to PPT3, which are equivalent to 0.046-0.123 mg Hg•L-1. Different dust extracts among the different sampling sites showed varying toxicity to PPT3. This study provides some important information to understand the potential health risk from different indoor environment using a rapid bioluminescence assay.

An Intact Cell Bioluminescence-Based Assay for the Simple and Rapid Diagnosis of Urinary Tract Infection.

Urinary tract infection (UTI) is one of the most common infections, accounting for a substantial portion of outpatient hospital and clinic visits. Standard diagnosis of UTI by culture and sensitivity can take at least 48 h, and improper diagnosis can lead to an increase in antibiotic resistance following therapy. To address these shortcomings, rapid bioluminescence assays were developed and evaluated for the detection of UTI using intact, viable cells of Photobacterium mandapamensis USTCMS 1132 or previously lyophilized cells of Photobacterium leiognathi ATCC 33981™. Two platform technologies-tube bioluminescence extinction technology urine (TuBETUr) and cellphone-based UTI bioluminescence extinction technology (CUBET)-were developed and standardized using artificial urine to detect four commonly isolated UTI pathogens-namely, Escherichia coli, Proteus mirabilis, Staphylococcus aureus, and Candida albicans. Besides detection, these assays could also provide information regarding pathogen concentration/level, helping guide treatment decisions. These technologies were able to detect microbes associated with UTI at less than 105 CFU/mL, which is usually the lower cut-off limit for a positive UTI diagnosis. Among the 29 positive UTI samples yielding 105-106 CFU/mL pathogen concentrations, a total of 29 urine specimens were correctly detected by TuBETUr as UTI-positive based on an 1119 s detection window. Similarly, the rapid CUBET method was able to discriminate UTIs from normal samples with high confidence (p ≤ 0.0001), using single-pot conditions and cell phone-based monitoring. These technologies could potentially address the need for point-of-care UTI detection while reducing the possibility of antibiotic resistance associated with misdiagnosed cases of urinary tract infections, especially in low-resource environments.

Efficient degradation of diclofenac by LaFeO3-Catalyzed peroxymonosulfate oxidation---kinetics and toxicity assessment.

Diclofenac was frequently found in various waters, indicating conventional wastewater treatment methods ineffective in its removal. In this study, LaFeO3 (LFO) was synthesized and its catalytic activity of LFO as the activator of different oxidants such as persulfate (PS), hydrogen peroxide and peroxylmonosulfate (PMS) was evaluated in terms of DCF degradation. The influence of calcination temperature was examined on the catalytic activity of LFO. The effects of various parameters including pH levels, PMS concentration, LFO dose and initial DCF concentration were investigated on DCF degradation rate. The marginal effects of PMS concentration and LFO dose were compared. Langmuir-Hinshelwood (LH) model was used to quantitatively describe DCF degradation reaction in LFO/PMS system. The two constants, k (Limiting reaction rate at maximum coverage) and K (Equilibrium adsorption constant), were determined on the basis of LH model. The performance of LFO/PMS process was also estimated in the presence of various inorganic anions. The potential toxicity of LFO and PMS were evaluated using phytoplankton and the toxicity evolution during DCF degradation was also investigated using luminescent bacteria. This contribution provides a basic study regarding the potential application of heterogeneous PMS activation by perovskite LFO for both DCF removal and toxicity elimination.

Acute toxicity assessment of explosive-contaminated soil extracting solution by luminescent bacteria assays.

Explosive-contaminated soil is harmful to people's health and the local ecosystem. The acute toxicity of its extracting solution was tested by bacterial luminescence assay using three kinds of luminescent bacteria to characterize the toxicity of the soil. An orthogonal test L 16 (45) was designed to optimize the soil extracting conditions. The optimum extracting conditions were obtained when the ultrasonic extraction time, ultrasonic extraction temperature, and the extraction repeat times were 6 h, 40 °C, and three, respectively. Fourier transform infrared spectroscopy (FTIR) results showed that the main components of the contaminated soil's extracting solution were 2,4-dinitrotoluene-3-sulfonate (2,4-DNT-3-SO3-); 2,4-dinitrotoluene-5-sulfonate (2,4-DNT-5-SO3-); and 2,6-dinitrotoluene (2,6-DNT). Compared with Photobacterium phosphoreum and Vibrio fischeri, Vibrio qinghaiensis sp. Nov. is more suitable for assessing the soil extracting solution's acute toxicity. Soil washing can remove most of the contaminants toxic to luminescent bacterium Vibrio qinghaiensis sp. Nov., suggesting that it may be a potential effective remediation method for explosive-contaminated soil.

iCar-PseCp: identify carbonylation sites in proteins by Monte Carlo sampling and incorporating sequence coupled effects into general PseAAC.

Carbonylation is a posttranslational modification (PTM or PTLM), where a carbonyl group is added to lysine (K), proline (P), arginine (R), and threonine (T) residue of a protein molecule. Carbonylation plays an important role in orchestrating various biological processes but it is also associated with many diseases such as diabetes, chronic lung disease, Parkinson's disease, Alzheimer's disease, chronic renal failure, and sepsis. Therefore, from the angles of both basic research and drug development, we are facing a challenging problem: for an uncharacterized protein sequence containing many residues of K, P, R, or T, which ones can be carbonylated, and which ones cannot? To address this problem, we have developed a predictor called iCar-PseCp by incorporating the sequence-coupled information into the general pseudo amino acid composition, and balancing out skewed training dataset by Monte Carlo sampling to expand positive subset. Rigorous target cross-validations on a same set of carbonylation-known proteins indicated that the new predictor remarkably outperformed its existing counterparts. For the convenience of most experimental scientists, a user-friendly web-server for iCar-PseCp has been established at http://www.jci-bioinfo.cn/iCar-PseCp, by which users can easily obtain their desired results without the need to go through the complicated mathematical equations involved. It has not escaped our notice that the formulation and approach presented here can also be used to analyze many other problems in computational proteomics.

Ecological risk assessment associated to the removal of endocrine-disrupting parabens and benzophenone-4 in wastewater treatment.

The occurrence of four widely used and endocrine disrupting parabens (PBs) (methylparaben, propylparaben, butylparaben and benzylparaben) and a polar UV filter (benzophenone-4) were determined in influent and effluent wastewater from the 19 major wastewater treatment plants (WWTPs) of Catalonia, Spain. For their analysis an on-line solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS) method was developed and validated. Laboratory analysis revealed high levels for both PBs and BP4, with maximum concentrations of 5700ngL(-1) and 1806ngL(-1), respectively, in influent samples, and 137ngL(-1) and 1080ngL(-1), respectively in effluent wastewaters. Removal rates (RE%) for the target compounds in each WWTPs were calculated. RE% for parabens were almost 100%, whereas for BP4 values where in the range 5-91%. The half-life time (t1/2), hydraulic retention time (HRT), and annual mass load (ML) for each facility was estimated. Results indicated that there was no clear influence of HRT on the RE% of BP4. MLs for BP4 were in the range 0.9-110.1kgy(-1), with the highest values in the most populated areas. Finally, a risk assessment, estimated in terms of hazard quotients (HQs), was carried out for aquatic biota. HQs for the target compounds in effluent wastewaters indicated a negligible effect, whereas for some influent wastewaters' HQs pointed out that some species are at risk.

Acute toxicity of benzophenone-type UV filters for Photobacterium phosphoreum and Daphnia magna: QSAR analysis, interspecies relationship and integrated assessment.

The hazardous potential of benzophenone (BP)-type UV filters is becoming an issue of great concern due to the wide application of these compounds in many personal care products. In the present study, the toxicities of BPs to Photobacterium phosphoreum and Daphnia magna were determined. Next, density functional theory (DFT) and comparative molecular field analysis (CoMFA) descriptors were used to obtain more detailed insight into the structure - activity relationships and to preliminarily discuss the toxicity mechanism. Additionally, the sensitivities of the two organisms to BPs and the interspecies toxicity relationship were compared. Moreover, an approach for providing a global index of the environmental risk of BPs to aquatic organisms is proposed. The results demonstrated that the mechanism underlying the toxicity of BPs to P. phosphoreum is primarily related to their electronic properties, and the mechanism of toxicity to D. magna is hydrophobicity. Additionally, D. magna was more sensitive than P. phosphoreum to most of the BPs, with the exceptions of the polyhydric BPs. Moreover, comparisons with published data revealed a high interspecies correlation coefficient among the experimental toxicity values for D. magna and Dugesia japonica. Furthermore, hydrophobicity was also found to be the most important descriptor of integrated toxicity. This investigation will provide insight into the toxicity mechanisms and useful information for assessing the potential ecological risk of BP-type UV filters.

Visible-light-mediated Sr-Bi2O3 photocatalysis of tetracycline: kinetics, mechanisms and toxicity assessment.

Photodegradation of tetracycline (TC) was investigated in aqueous solution by visible-light-driven photocatalyst Sr-doped ß-Bi2O3 (Sr-Bi2O3) prepared via solvothermal synthesis. The decomposition of TC by Sr-Bi2O3 under visible light (λ>420nm) irradiation followed pseudo-first-order kinetics, and the removal ratio reached 91.2% after 120min of irradiation. Sr-Bi2O3 photocatalysis is able to break the naphthol ring of TC which decomposes to m-cresol via dislodging hydroxyl group step by step by photogenerated electron. This mechanism was verified by electron spin resonance measurement, the addition of radical scavengers and the intermediate product analysis, indicating that the photogenerated electron acts as a reductant and can be the key to the degradation process. In contrast, in TiO2 photocatalysis the naphthol ring is broken via oxidation by hydroxyl radical, while in direct photolysis the ring remains intact. In addition, the toxicity of photodegradation products was analyzed by bioluminescence inhibition. After 120min of irradiation by Sr-Bi2O3, the toxicity decreases by 90.6%, which is more substantial than direct photolysis (70%) and TiO2 photocatalysis (80%), indicating that the Sr-Bi2O3 photocatalysis is more eco-friendly than the other two methods.

[Acute toxicity of antibiotics and anaerobic digestion intermediates in pharmaceutical wastewaters].

In order to determine the toxicity of antibiotics and anaerobic digestion intermediates on anaerobic treatment of pharmaceutical wastewaters containing antibiotics, the single and joint toxicities of some antibiotics and intermediates to Photobacterium phosphoreum were tested by using the 15-min half inhibitory concentration (15 min-IC50) at pH = 7.00 +/- 0.05. The results showed that the 15 min-IC50 of ethanol, acetate, propionate and butyrate were 19.40, 20.71, 10.47 and 12.17 g x L(-1), respectively, which indicated that the toxicity descended in the order of propionate, butyrate, ethanol and acetate. The 15 min-IC50 of Amoxicillin, Kanamycin, Lincomycin and Ciprofloxacin were 3.99, 5.11, 4.32 and 5.63 g x L(-1), respectively, so the toxicity descended in the order of Amoxicillin, Lincomycin, Kanamycin and Ciprofloxacin. Using equal effect mixing method, the joint toxicity of four anaerobic digestion intermediates, the four intermediates together with Amoxicillin, Ciprofloxacin, Kanamycin, Lincomycin individually and all together were investigated, which demonstrated that the first three interactions were additive and the last three were synergistic. The observations have laid a foundation for control and optimization of anaerobic biotechnology for pharmaceutical wastewater containing antibiotics.

Toxicity-based assessment of the treatment performance of wastewater treatment and reclamation processes.

The reclamation and reuse of wastewater is one of the possible ways to relieve the serious fresh water resource crisis in China. Efficient reclamation treatment technologies ensure the safe reuse of reclaimed water. In order to screen out and evaluate technologies appropriate for reclamation treatment, a great deal of efforts have been brought to bear. In the present study, a toxicity-based method including a Photobacterium phosphoreum test for acute toxicity and SOS/umu test for genotoxicity, accompanied by the traditional physicochemical parameters DOC (dissolved organic carbon) and UV254 (absorbance at 254 nm), was used to measure the treatment performance of different reclamation processes, including the anaerobic-anoxic-oxic biological process (A2O) and subsequent physical/chemical reclamation processes (ultrafiltration, ozonation, chlorination). It was found that for the secondary effluent after the A2O process, both the toxicity and physicochemical indices had greatly decreased compared with those of the influent. However, chemical reclamation processes such as ozonation and chlorination could possibly raise toxicity levels again. Fortunately, the toxicity elevation could be avoided by optimizing the ozone dosage and using activated carbon after ozonation. It was noted that by increasing the ozone dosage to 10 mg/L and employing activated carbon with more than 10 min hydraulic retention time, toxicity elevation was controlled. Furthermore, it was shown that pre-ozonation before activated carbon and chlorination played an important role in removing organic compounds and reducing the toxicity formation potential. The toxicity test could serve as a valuable tool to evaluate the performance of reclamation processes.

Ecotoxicological assessment of leachates from MSWI bottom ashes.

In this paper, chemical and ecotoxicological data of leachates from bottom ashes collected in different Municipal Solid Waste Incinerators (MSWI) are shown. The bottom ashes were collected in Belgium (three incinerators--samples B1 to B3), France, Germany, Italy and United Kingdom (one incinerator in each country--samples F1, D1, I1 and UK1, respectively). Both chemical and ecotoxicological characterizations of leachates were done on the framework of the European Directive 91/689/EEC and the European Council Decision 94/904/EC. This work was carried out under the European project called Valomat, which was supported by the European Commission through Brite-Euram III program. Twenty-one inorganic parameters were analyzed. The ecotoxicological assays were done under standard laboratory conditions, using the bacterium Photobacterium phosphoreum, the freshwater alga Pseudokirchneriella subcapitata, the crustacean Daphnia magna and the vegetable Lactuca sativa. Chemical data varied from sample to sample. Similar results were obtained in biological assays. The samples can be classified as ecotoxic/hazardous according to the French proposal for a Criterion and Evaluation Methods of Waste Ecotoxicity (CEMWE) and the German regulation on Hazardous Waste Classification (HWC). However, samples B1, B2, B3 and D1 comply the maximum limits for direct valorization category defined in the French Classification of Bottom Ashes based on their Polluting Potential (CBAPP). Sample B1 presented the lowest level of ecotoxicity, being considered as the most interesting to be used in the development of new materials for civil engineering works.

A novel and sensitive test for rapid determination of water toxicity.

The performance of a novel, rapid, and sensitive test for detecting chemical toxicants in water is described in this article. The bioassay utilizes a highly sensitive variant of the luminescent bacterium Photobacterium leiognathi that allows the detection in water at levels below milligrams per liter of diverse groups of toxicants, including heavy metals, pesticides, PCBs, polycyclic aromatic hydrocarbons, and fuel traces. For most toxic agents reported in this study, the new assay was markedly more sensitive than the Microtox(trade mark) Vibrio fischeri assay according to the bacterial bioluminescence toxicity data reported in the literature. Additional features of the new bioassay include the ability to discriminate between cationic heavy metals and organic toxicants and the option of being run at ambient temperatures (18 degrees C-27 degrees C), thereby enabling on-site testing with low-cost luminometers. In addition, the stability of the freeze-dried bacterial reagent preparation at ambient temperatures precludes the need for refrigeration or freezing during shipment, which contributes to further reducing overall operational costs.

A case study of logistic QSAR modeling methods and robustness tests.

Acute toxicity (15-min EC(50)) determination of 16 substituted naphthalene compounds to Photobacterium phosphoreum was undertaken according to the standard procedures, while the effects of molecular structures of selected compounds on their toxicity to test microorganisms were logistically conducted using the quantitative structure-activity relationship (QSAR) technique. The relationship was developed as -log EC(50)=5.5916 (+/-0.1189)-7.4893(+/-0.4900)qH(+)+0.7771(+/-0.0619E(lumo)+0.0088(+/-0.0009)alpha(N=16, R(2)(adj)=0.9698, SE=0.0892, P=0.0000). The cluster analyses of individual structure descriptors as well as the quality control chart and Monte Carlo simulation test indicated that the prediction model was reasonable and robust even if tested with several different methods. Furthermore, the quantum chemical parameters entering into the QSAR model were used to discuss the possible toxicity pathways, and the results revealed that the selected compounds were reactive and their toxicity behaviors were complex processes containing physical partition stages as well as biochemical reaction stages.

An ecotoxic risk assessment of residue materials produced by the plasma pyrolysis/vitrification (PP/V) process.

Plasma is the fourth state of matter, following the three states of solid, liquid and gas. Experience has amply demonstrated that solids exposed to the oxygen-deficient plasma flame are converted to liquid, and liquid exposed to the same flame is converted to gas. A low amount of vitrified solid residue material usually remains at the end of this process. Plasma pyrolysis/vitrification (PP/V) has been demonstrated as a safe, efficient, cost-effective technology for the treatment of wastes, including hazardous wastes. Besides the low amounts of gaseous byproducts that PP/V produces, the solid vitrified residue presents a low leachability of pollutants. Studies have been performed in many countries in order to assess the leachability of chemical substances. But from the results of identified studies, none has reported results on the ecotoxicological properties of the leachates. The aim of this study was to contribute to the assessment of ecotoxic risk of four different vitrified materials. Vitrified samples of contaminated soils, municipal solid wastes, and incinerator bottom ashes were submitted to the European leaching pre-standard test number prEN 12457-2. The leachates were analyzed for 22 chemical parameters. The biological characterization comprised the assessment of bioluminescence inhibition of Photobacterium phosphoreum bacterium, growth inhibition of Pseudokirchneriella subcapitata algae and the germination inhibition of Lactuca sativa vegetable. The chemical and ecotoxicological results were analyzed according to the French proposal of Criteria on the Evaluation Methods of Waste Toxicity (CEMWT) and a Toxicity Classification System (TCS). The chemical and ecotoxicological results indicated a low leachability of pollutants and a low toxicity level of leachates. All samples studied were as below the TCS class 1 level (no significant toxicity observed) and as non-ecotoxic for CEMWT. Therefore, the environmental ecotoxic risk of the analyzed vitrified samples was determined to be very low.

Assessment of a magnetic bead-EIA based kit for rapid diagnosis of fish pasteurellosis.

The accuracy of the magnetic beads-EIA based BIONOR AQUAEIA-Pp kit for the rapid diagnosis of pasteurellosis was evaluated. The kit reacted with all the Photobacterium damselae subsp. piscicida strains included in this study, with a detection limit of 10(4) bacteria/ml. However, non-specific reactions were observed with isolates of Ph. damselae subsp. damselae or Ph. histaminum when the bacterial concentration was high (10(9)-10(10) bacteria/ml). Similar findings in specificity and sensitivity were observed when the kit was applied to experimentally infected fish tissues. However, since those bacterial species are not usually found in the fish species susceptible to pasteurellosis, the AQUAEIA kit appears applicable for a rapid screening of the disease. In addition, when the kit was utilized to analyze cultured populations of seabream, it allowed the detection of the pathogen, not only in individuals affected by the disease, but also in asymptomatic carrier fish. Furthermore, the positive detection of Ph. damselae subsp. piscicida in broodstock gonads, seminal, and ovaric fluids, and also in eggs indicated the possibility of vertical transmission of pasteurellosis.

Ecotoxicological assessment of ashes and particulate matter from fluidized bed combustion of coal.

Toxicity of ash and particulate matter from the gases generated in coal fluidized bed combustion (FBC) has been determined by the Microtox assay according to the standard leachate procedure. Results are compared with the polycylic aromatic hydrocarbon (PAH) content of the particulate matter, which was determined by fluorescence spectroscopy. Although PAHs are considered highly toxic compounds, the low ecotoxicity values obtained could be explained by the low solubility of the compounds in water. The Microtox assay may underestimate the toxicity of water-insoluble compounds unless they are previously extracted with an organic solvent. Nevertheless, this type of assay can be very useful for measuring the potential toxicity of residues when exposed to water sources such as rain water and the risk of the components being dissolved and transported by runoff water.