Influence of extracellular polymeric substances (EPS) on Cd adsorption by bacteria

The role of extracellular polymeric substances (EPS) in Cd adsorption by Bacillus subtilis and Pseudomonas putida was investigated using a combination of batch adsorption experiments, potentiometric titrations, Fourier transform infrared spectroscopy (FTIR). An increased adsorption capacity of Cd was observed for untreated bacteria relative to that for EPS-free bacteria. Surface complexation modeling of titration data showed the similar pK_a values of functional groups (carboxyl, phosphate and hydroxyl) between untreated and EPS-free bacteria. However, site concentrations on the untreated bacteria were found to be higher than those on the EPS-free bacteria. FTIR spectra also showed that no significant difference in peak positions was observed between untreated and EPS-free bacteria and carboxyl and phosphate groups were responsible for Cd adsorption on bacterial cells. The information obtained in this study is of fundamental significance for understanding the interaction mechanisms between heavy metals and biofilms in natural environments.     

Function of bacterial cells and their exuded extracellular polymeric substances (EPS) in virus removal by red soils

The potential influence of autochthonous microorganisms on virus fate in soil is usually determined through extreme conditions of sterilization vs. nonsterilization; however, the relative importance of microbial cells and their exudates remains unclear. In this study, bacterial cells (cell) were harvested, and their exuded extracellular polymeric substances (EPS) were extracted from three strains of bacteria, namely, Gram-negative bacteria Pseudomonas putida and Pseudomonas aeruginosa as well as Gram-positive bacterium Bacillus subtilis. This study aimed to evaluate virus removal in solutions in the presence of cell, EPS, and their combination (cell/EPS), as well as to investigate how their presence affects virus removal efficiencies by four red soils based on batch experiments. Results showed that virus removal percentage in solutions ranged from 11 to 23 in the presence of cells only and from 12 to 15 in the presence of EPS only. The removal percentage in the combined cell/EPS treatment can be estimated by summing the results achieved by the cell and EPS treatments, separately. Meanwhile, cell presence had a negligible effect on virus removal by red soils. EPS and combined cell/EPS significantly reduced virus removal by 20 to 69 % and 16 to 50 %, respectively, which indicated that EPS served a dominant function in reducing virus removal. This study clearly demonstrated that the prediction of virus removal by red soils must consider the effect of bacteria, especially those producing large quantities of EPS, which can be responsible for the underestimation of viral load in certain studies.     

Characterization of extracellular polymeric substances (EPS) from periphyton using liquid chromatography-organic carbon detection–organic nitrogen detection (LC-OCD-OND)

A protocol was developed to extract, fractionate, and quantitatively analyze periphyton extracellular polymeric substances (EPS), which obtains both information on the molecular weight (M _r) distribution and protein and polysaccharide content. The EPS were extracted from freshwater periphyton between July and December 2011. Organic carbon (OC) compounds from different EPS extracts were analyzed using liquid chromatography-organic carbon detection–organic nitrogen detection (LC-OCD-OND), and total protein and polysaccharide content were quantified. Four distinct OC fractions, on the basis of M _r, were identified in all extracts, corresponding to high M _r biopolymers (≥80–4 kDa), degradation products of humic substances (M _r not available), low M _r acids (10–0.7 kDa), and small amphiphilic/neutral compounds (3–0.5 kDa). Low C/N ratios (4.3?±?0.8) were calculated for the biopolymer fractions, which represented 16–38 % of the measured dissolved organic carbon (DOC), indicating a significant presence of high M _r proteins in the EPS. Protein and polysaccharide represented the two major components of EPS and, when combined, accounted for the measured DOC in extracts. Differences in specific OC fractions of EPS extracts over the course of the study could be quantified using this method. This study suggests that LC-OCD-OND is a new valuable tool in EPS characterization of periphyton.     

Pb remobilization by bacterially mediated dissolution of pyromorphite Pb5(PO4)3Cl in presence of phosphate-solubilizing Pseudomonas putida

Remediation of lead (Pb)-contaminated sites with phosphate amendments is one of the best studied and cost-effective methods for in situ immobilization. In this treatment, a very stable mineral, pyromorphite Pb₅(PO₄)₃Cl, is formed. Several studies propose to improve this treatment method with the addition of phosphate-solubilizing bacteria (PSB). The effect of bacteria on solubilization of pyromorphite is unknown. In this study, the effect of the soil microorganisms on the stability of pyromorphite Pb₅(PO₄)₃Cl has been investigated in a set of batch solution experiments. The mineral was reacted with Pseudomonas putida, a common soil microorganism. Dissolution of pyromorphite was enhanced by the presence of P. putida, resulting in an elevated Pb concentration in the solution. This occurred even when the bacteria were provided with an additional source of phosphate in the solution. Pyromorphite has been shown to be a potential source of nutrient phosphorus for common soil bacteria. Thus, the use of PSB in remediation treatments of Pb contaminated sites may have adverse long-term impacts on Pb immobilization. Conscious phosphate management is suggested for long-term sustainability of the in situ Pb immobilization by pyromorphite formation.     

Bioadsorption and bioaccumulation of chromium trivalent in Cr(III)-tolerant microalgae: A mechanisms for chromium resistance

Anthropogenic activity constantly releases heavy metals into the environment. The heavy metal chromium has a wide industrial use and exists in two stable oxidation states: trivalent and hexavalent. While hexavalent chromium uptake in plant cells has been reported that an active process by carrying essential anions, the cation Cr(III) appears to be taken up inactively. Dictyosphaerium chlorelloides (Dc1M), an unicellular green alga is a well-studied cell biological model organism. The present study was carried out to investigate the toxic effect of chromium exposures on wild-type Cr(III)-sensitive (Dc1M^wt) and Cr(III)-tolerant (Dc1M^Cr(III)R30) strains of these green algae, and to determine the potential mechanism of chromium resistance. Using cell growth as endpoint to determine Cr(III)-sensitivity, the IC₅₀₍₇₂₎ values obtained show significant differences of sensitivity between wild type and Cr(III)-tolerant cells. Scanning electron microscopy (SEM) showed significant morphological differences between both strains, such as decrease in cell size or reducing the coefficient of form; and transmission electron microscopy (TEM) revealed ultrastructural changes such as increased vacuolization and cell wall thickening in the Cr(III)-tolerant strain with respect to the wild-type strain. Energy dispersive X-ray analysis (SEM/XEDS) revealed that Cr(III)-tolerant D. chlorelloides cells are able to accumulate considerable amounts of chromium distributed in cell wall (bioadsorption) as well as in cytoplasm, vacuoles, and chloroplast (bio-accumulation). Morphological changes of Cr(III)-tolerant D. chlorelloides cells and the presence of these electron-dense bodies in their cell structures can be understood as a Cr(III) detoxification mechanism.     

A new bioseed for determination of wastewater biodegradability: analysis of the experimental procedure

A new bioassay proposed in the patent P201300029 was applied to a pre-treated wastewater containing a mixture of commercial pesticides to simulate a recalcitrant industrial wastewater in order to determine its biodegradability. The test uses a mixture of standardized inoculum of the lyophilized bacteria Pseudomonas putida with the proper proportion of salts and minerals. The results highlight that biodegradation efficiency can be calculated using a gross parameter (chemical oxygen demand (COD)) which facilitates the biodegradability determination for routine water biodegradability analysis. The same trend was observed throughout the assay with the dehydrated and fresh inoculums, and only a difference of 5 % in biodegradation efficiency (E _f) was observed. The obtained results showed that the P. putida biodegradability assay can be used as a commercial test with a lyophilized inoculum in order to monitor the ready biodegradability of an organic pollutant or a WWTP influent. Moreover, a combination of the BOD₅/COD ratio and the P. putida biodegradability test is an attractive alternative in order to evaluate the biodegradability enhancement in water pre-treated with advanced oxidation processes (AOPs).     

Relationship between chromium(VI) resistance and extracellular polymeric substances (EPS) concentration by some cyanobacterial isolates

BACKGROUND, AIM, AND SCOPE: Chromium(VI) resistance and its association with extracellular polymeric substance (EPS) concentration in cyanobacteria was investigated. Increased EPS concentration was associated with Cr(VI) resistance. The most resistant isolate, Chroococcus sp. H(4), secreted the most EPS (427 mg/L). MATERIALS AND METHODS: EPS concentration of the two most resistant isolates (Chroococcus sp. H(4) and Synechocystis sp. S(63)) was investigated following exposure to 15 and 35 ppm Cr(VI). The composition of EPS produced by Chroococcus sp. H(4) following exposure to 10 ppm Cr(VI) was analyzed using high-performance liquid chromatography. Control EPS was composed of glucose (99%) and galactronic acid (1%); in the presence of 10 ppm Cr(VI), EPS composition changed to glucose (9%), xylose (75%), rhamnose (14%), and galacturonic acid (2%). RESULTS AND DISCUSSION: Results indicated that (1) exposure to elevated concentrations of Cr(VI) affected the composition of EPS produced by Chroococcus sp. H(4), and (2) there was a correlation between Cr(VI) resistance and EPS concentration in some cyanobacteria.     

Quantification of the interactions between Ca,Hg and extracellular polymeric substances (EPS) of sludge

The interactions between metals (Ca²⁺ and Hg²⁺) and extracellular polymeric substances (EPS) extracted from the aerobic and anaerobic sludge in wastewater treatment reactors were investigated using a combination of zeta potential measurement and 3-dimensional excitation–emission matrix (EEM) fluorescence spectroscopy with parallel factor (PARAFAC) analysis. Results show that Ca²⁺ had no substantial effects on the EEM fluorescence spectra of the EPS, but their zeta potentials increased with the increasing Ca²⁺ dosage. However, Hg²⁺ had a significant effect on the EEM fluorescence spectra of the EPS, while their zeta potentials seemed not to be affected by the dose of Hg²⁺. The interactions between Hg²⁺ and EPS were elucidated using the fluorescence quenching with PARAFAC analysis, while the interactions between Ca²⁺ and EPS were evaluated by the zeta potential technique. The binding constants for Hg²⁺ and EPS were two orders of magnitude higher than those for Ca²⁺ and EPS, suggesting that the binding mechanisms between Ca²⁺ and EPS were different from those between Hg²⁺ and EPS. The results might be useful for understanding the roles of EPS in bacterial self-protection against heavy metals and the aggregate formation mechanisms through ionic bridging interactions.     

Evaluation of bioaugmentation and biostimulation on arsenic remediation in soil through biovolatilization

Arsenic (As) removal through microbially driven biovolatilization can be explored as a potential method for As bioremediation. However, its effectiveness needs to be improved. Biostimulation with organic matter amendment and bioaugmentation with the inoculation of genetic engineered bacteria could be potential strategies for As removal and site remediation. Here, the experiments were conducted to evaluate the impacts of rice straw and biochar amendment, inoculation of genetic engineered Pseudomonas putida KT2440 (GE P. putida) with high As volatilization activity, on microbial mediated As volatilization and removal from three different arseniferous soils. In general, the addition of rice straw (5%) significantly enhanced As methylation and volatilization in comparison with corresponding non-amended soils. Biochar amendments and inoculation of the GE P. putida increased As methylation and volatilization, respectively, but less than that of rice straw addition. The effectiveness of As volatilizations are quite different in the various paddy soils. The combined amendments of rice straw and GE P. putida exhibited the highest As removal efficiency (483.2 μg/kg/year) in Dayu soil, with 1.2% volatilization of the total As annually. The highest water-soluble As concentration (0.73 mg/kg) in this soil could be responsible for highest As volatilization besides the rice straw and bacteria in this soil.     

Modification of exopolysaccharide composition and production by three cyanobacterial isolates under salt stress

Background, aim, and scope  

Polysaccharides are renewable resources representing an important class of polymeric materials of biotechnological interest, offering a wide variety of potentially useful products to mankind. Exopolysaccharides (EPSs) of microbial origin with a novel functionality, reproducible physico-chemical properties, stable cost and supply, became a better alternative to polysaccharides of algal origin. EPSs are believed to protect bacterial cells from desiccation, heavy metals or other environmental stresses, including hostimmune responses, and to produce biofilms, thus enhancing the cells chances of colonising special ecological niches. One of the most important stress factor is salt stress for microorganisms. The present investigation is aimed to determine correlation between salt resistance and EPS production by three cyanobacterial isolates (Synechocystis sp. BASO444, Synechocystis sp. BASO507 and Synechocystis sp. BASO511). It is also aimed to investigate the effect of salt concentrations on EPS production by cyanobacteria and effect of salt on monosaccharide composition of EPS.     

Metal binding properties of extracellular polymeric substances extracted from anaerobic granular sludges

Extracellular polymeric substances (EPS) were extracted from four anaerobic granular sludges with different procedures to study their involvement in biosorption of metallic elements. EPS extracts are composed of closely associated organic and mineral fractions. The EPS macromolecules (proteins, polysaccharides, humic-like substances, nucleic, and uronic acids) have functional groups potentially available for the binding of metallic elements. The acidic constants of these ionizable groups are: pK _a1 (4–5) corresponding to the carboxyl groups; pK _a2 (6–7) corresponding to the phosphoric groups; pK _a3 (8–10) and pK _a4 (≈10) corresponding to the phenolic, hydroxyl, and amino groups. The polarographic study confirms the higher affinity of the EPS to bind to lead than to cadmium. Moreover, the binding of these metallic compounds with the EPS is a mix of several sorption mechanisms including surface complexation, ion exchange, and flocculation. Inorganic elements were found as ions linked to organic molecules or as solid particles. The mineral fraction affects the binding properties of the EPS, as the presence of salts decreases the EPS binding ability. Calcite and apatite particles observed on SEM images of EPS extracts can also sorb metallic elements through ion exchange or surface complexation.     

Investigation of the removal mechanism of Cr(VI) in groundwater using activated carbon and cast iron combined system

Zero-valent iron (Fe⁰) has been widely used for Cr(VI) removal; however, the removal mechanisms of Cr(VI) from aqueous solution under complex hydrogeochemical conditions were poorly understood. In this research, the mixed materials containing cast iron and activated carbon were packed in columns for the treatment of aqueous Cr(VI)-Cr(III) in groundwater with high concentration of Ca²⁺, Mg²⁺, HCO₃ ⁻, NO₃ ⁻, and SO₄ ²⁻. We investigate the influences of those ions on Cr(VI) removal, especially emphasizing on the reaction mechanisms and associated precipitations which may lead to porosity loss by using X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) techniques. The results show that the precipitations accumulated on the material surface were (Fe/Cr) (oxy)hydroxide, mixed Fe(III)-Cr(III) (oxy)hydroxides, Fe₂O₃, CaCO₃, and MgCO₃. During these reactions, the Cr(VI) was reduced to Cr(III) coupled with the oxidated Fe⁰ to Fe(II) through the galvanic corrosion formed by the Fe⁰-C and/or the direct electron transfer between Fe⁰ and Cr(VI). In addition, Cr(VI) could be reduced by aqueous Fe(II), which dominated the whole removal efficiency. The primary aqueous Cr(III) was completely removed together with Cr(III) reduced from Cr(VI) even when Cr(VI) was detected in the effluent, which meant that the aqueous Cr(III) could occupy the adsorption sites. In general, the combined system was useful for the Cr(VI)-Cr(III) treatment based on galvanic corrosion, and the hardness ions had a negative effect on Cr(VI) removal by forming the carbonates which might promote the passivation of materials and decrease the removal capacity of the system.

     

Human cell death in relation to DNA damage after exposure to the untreated and biologically treated pharmaceutical wastewater

Among all the pharmaceutical drugs that contaminate the environment, antibiotics occupy an important place due to their high consumption rates in both veterinary and human medicine. The present study examined the ability of Pseudomonas putida to grow on the antibiotic wastewater, currently expanding in Tunisia, containing amoxicillin and cefadroxil. P. putida was very efficient to grow quickly in pharmaceutical wastewater (PW) and in reducing the total dissolved solids to 80.1 %. Cytotoxicity of PW, before and after biodegradation with P. putida mt-2, was evaluated in vitro, using the MTT assay, against four human tumor cell lines such as A549 (lung cell carcinoma), HCT15 (colon cell carcinoma), MCF7 (breast adenocarcinoma), and U373 (glioma cell carcinoma). The PW reduced all human cell lines viability in a dose-dependent manner. This activity was very remarkable against U373 cell line. For this reason, we have tested the genotoxicity of PW using comet assay for quantification of DNA fragmentation. In fact, PW has statistically significant (p?<?0.001) influence on DNA. Indeed, the percentage of genotoxicity was 66.87 and 87.5 %, after 24 and 48 h of treatment, respectively. However, cytotoxicity and genotoxicity decreased strongly when tested the PW obtained after incubation with P. putida mt-2. Our results indicate that P. putida is a promising and improved alternative to treating industrial-scale effluent compared to current chemical treatment procedures used by the industrials.     

Assessment of a process to degrade metal working fluids using Pseudomonas stutzeri CECT 930 and indigenous microbial consortia

The development of a novel biological process to treat metal working fluids (MWFs)-containing effluents at bioreactor scale was pursued in this work. The bacteria Pseudomonas stutzeri CECT 930 was investigated for the first time as an alternative agent for MWF degradation. An adequate medium design and mixing and aeration system, as well as an appropriate microorganism proved to be crucial for reaching high levels of degradation by P. stutzeri and by an indigenous consortium (about 70% and 50% of reduction in total petroleum hydrocarbon content in less than 2 wk, respectively). Additionally, as there is no information in literature trying to kinetically characterize an MWF-polluted effluent degradation process, all the experimental data were fitted to logistic and Luedeking and Piret models, that allowed to elucidate the growth-associated character of the biodegradation process.     

Effect of fungicides on plant growth promoting activities of phosphate solubilizing Pseudomonasputida isolated from mustard (Brassica compestris) rhizosphere

This study was navigated to examine the effects of fungicide-stress on the activities of plant-growth-promoting rhizobacterium Pseudomonasputida with inherent phosphate solubilizing activity. The fungicide-tolerant and phosphate solubilizing P.putida strain PS9 was isolated from the mustard rhizosphere and tentatively identified following standard morphological, physiological and biochemical tests. To further consolidate the identity of the strain PS9, the 16S rDNA sequence analysis was performed. Following the BLAST program, the strain PS9 was identified as P.putida. In the presence of the varying concentrations (0-3200 μg mL⁻¹; at a two fold dilution interval) of four fungicides of different chemical families (tebuconazole, hexaconazole, metalaxyl and kitazin) amended in minimal salt agar medium, the P.putida strain PS9 showed a variable tolerance levels (1400-3200 μg mL⁻¹) against the tested fungicides. The strain PS9 produced plant-growth-promoting (PGP) substances in significant amount in the absence of fungicides. In general, fungicides applied at the recommended, two and three times of the recommended rates, decreased the PGP attributes of P.putida the strain PS9 and affected the PGP activities in concentration-dependent manner. Fungicides at the recommended dose had minor reducing effect while the doses higher than the recommended dose significantly reduced the PGP activities (phosphate solubilization, salicylic acid, 2,3-dihydroxy benzoic acid, and indole-3-acetic acid production except exo-polysaccharides, hydrogen cyanate and ammonia production). Of the four fungicides, tebuconazole generally, showed maximum toxicity to the PGP activities of the strain PS9. This study inferred that fungicides must be examined in vitro for their possible adverse effects on soil micro flora before their application in agricultural fields. Moreover, the results also suggested the prerequisite of application of fungicide-tolerant PGPR strains as bioinoculants so that their PGP activities may not be suppressed under fungicide stress.     

Potential of the TCE-degrading endophyte Pseudomonas putida W619-TCE to improve plant growth and reduce TCE phytotoxicity and evapotranspiration in poplar cuttings

The TCE-degrading poplar endophyte Pseudomonas putida W619-TCE was inoculated in poplar cuttings, exposed to 0, 200 and 400 mg l⁻¹ TCE, that were grown in two different experimental setups. During a short-term experiment, plants were grown hydroponically in half strength Hoagland nutrient solution and exposed to TCE for 3 days. Inoculation with P. putida W619-TCE promoted plant growth, reduced TCE phytotoxicity and reduced the amount of TCE present in the leaves. During a mid-term experiment, plants were grown in potting soil and exposed to TCE for 3 weeks. Here, inoculation with P. putida W619-TCE had a less pronounced positive effect on plant growth and TCE phytotoxicity, but resulted in strongly reduced amounts of TCE in leaves and roots of plants exposed to 400 mg l⁻¹ TCE, accompanied by a lowered evapotranspiration of TCE. Dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA), which are known intermediates of TCE degradation, were not detected.     

Simultaneous reduction of Cr(VI) and oxidation of As(III) by Bacillus firmus TE7 isolated from tannery effluent

Hexavalent chromium [Cr(VI)] and arsenite [As(III)] are the most toxic forms of chromium and arsenic respectively, and reduction of Cr(VI) to Cr(III) and oxidation of As(III) to As(V) has great environmental implications as they affect toxicity and mobility of these toxic species. Bacillus firmus strain TE7, resistant to chromium and arsenic was isolated from tannery effluent. The strain exhibited ability to reduce Cr(VI) and oxidize As(III). It reduced 100 mg L^?1 Cr(VI) within 60 h in nutrient broth and oxidized 150 mg L^?1 As(III) within 10 h in minimal medium. It also completely reduced 15 mg L^?1 Cr(VI) and oxidized 50 mg L^?1 of As(III) simultaneously in minimal medium. To the best of our knowledge, this is the first bacterial strain showing simultaneous reduction of Cr(VI) and oxidation of As(III) and is a potential candidate for bioremediation of environments contaminated with these toxic metal species.     

Production of a monoclonal antibody and development of an immunoassay for detection of Cr(III) in water samples

In this study we report the production of a monoclonal antibody (Mab) specific for Cr(III)-chelate and the development of a competitive immunoassay for detection of Cr(III) in water samples. In the assay, the complete antigen (Cr(III)-ITCBE-BSA) was used as coating antigen, and Cr(III)-ITCBE as competitor competes with coating antigen to bind with Mab. Using this approach, the spiked water samples with Cr(III) were detected. The linear range of the detection was 0.7–12.4 ng mL⁻¹. The limit of the detection (LOD) was 0.51 ng mL⁻¹. The spiked results were also confirmed by ICP-MS, which showed a good correlation (R² = 0.997) between the two methods. The results indicated that the developed assay was reliable and suitable for the detection of Cr(III) in water samples.     

Alteration of in vitro and acute in vivo toxicity of textile dyeing wastewater after chemical and biological remediation

Introduction

Textile industry is one of the most common and essential sectors in Tunisia. However, the treatment of textile effluents becomes a university because of their toxic impacts on waters, soils, flora, and fauna.

Materials and methods

The aim of this work was to evaluate the ability of Pseudomonas putida mt-2 to decolorize a textile wastewater and to compare the biologic decolorization process to the chemical one currently used by the textile industry.

Results

P. putida exhibited a high decolorizing capacity of the studied effluent, compared to the coagulation?Cflocculation method with decolorization percentage of 86% and 34.5%, respectively. Genotoxicity of the studied effluent, before and after decolorization by P. putida mt-2, was evaluated in vitro, using the SOS chromotest, and in vivo, in mouse bone marrow, by assessing the percentage of cells bearing different chromosome aberrations compared to not treated mice. In addition, textile effluent statistically significant influenced acetylcholinesterase and butyrylcholinesterase activities and lipid peroxidation (p?P. putida is a promising and improved alternative to treating industrial scale effluent compared to current chemical decolorization procedures used by the Tunisian textile industry.