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Water pollution, especially caused due to the indiscriminate release of heavy metals as a result of anthropogenic activities is a major concern worldwide. Chromium, a heavy metal, regardless of its commercial importance has found to be a potent water pollutant. Chromium generally exist as hexavalent (Cr(VI)) and trivalent (Cr(III)) chromium in the environment. Cr(VI) is ascertained to be more toxic compared to Cr(III) and the former is identified as a carcinogen by the World Health Organisation (WHO). Some of the conventional methods currently available for chromium pollution mitigation are not cost effective and most importantly lead to secondary pollution in the form of sludge. Bioremediation is a promising alternative technique which is also ecofriendly. The bioremediation process utilises biological materials such as microorganisms and agricultural byproducts. Biosorption is a bioremediation process that is a surface related phenomenon involving adsorption of contaminant chromium ions onto the binding sites of the biosorbents. In addition to the efforts made to the remediation of chromium, continuous monitoring of chromium contaminant level in polluted water bodies becomes imperative. The present research study encompasses findings related to bioremediation and detection of chromium ions using bacterial cells. The first part of the dissertation involves studies pertaining to the bioremediation of chromium ions using different bacterial strains as biosorbent. For the study, bacterial strains procured from a microbial culture collection bank as well as those isolated from chromium polluted water samples collected from an industrial site were assessed for their ability to remediate chromium. The next aim of the study was to elucidate the mechanisms involved in the bioremediation of chromium ions by the bacterial cells for which the different characterisation methods such as, Fourier Transform Infrared (FTIR) spectroscopy, Energy Dispersive Spectroscopy (EDS), X-ray Photoelectron Spectroscopy (XPS) and zeta potential measurements which enabled to throw light on the reactions occurring at the bacterial cell surface-chromium solution interface. The later part of the study examines the capability of the bacterial strains used in the bioremediation studies as sensors for the detection of Cr(VI) and Cr(III) ions by adopting electroanalytical techniques, such as, Cyclic Voltammetry (CV) and Cathodic Stripping Voltammetry (CSV), wherein a microbe-modified Carbon Paste Electrode (CPE) was used as the working electrode in a typical three electrode electrochemical cell with Saturated Calomel Electrode (SCE) and platinum wire used as the reference and auxiliary electrodes respectively. The key objectives of the present study are as follows: (i) To study the bioremediation of Cr(VI) and Cr(III) ions present in aqueous solutions using two bacterial strains procured from a microbial culture collection bank as biosorbents. The bacterial strains used were Corynebacterium paurometabolum (Cp), a Gram positive bacterium and Citrobacter freundii (Cf), a Gram negative bacterium. The various factors affecting the biosorption process are to be investigated. (ii) To isolate and identify bacterial strains from water samples collected from chromium contaminated mining site in Sukinda, Odisha, India, by adopting appropriate microbiological and molecular biological procedures. (iii) To study the various factors affecting bioremediation of Cr(VI) using the mine isolates (Chromobacterium sp. (Cb) and Sphingopyxis sp. (Sp)) both Gram negative, as biosorbents. (iv) To elucidate the mechanisms adopted by the chosen bacterial cells in the bioremediation of chromium. (v) To develop an electrochemical-microbial sensor by modifying the Carbon Paste Electrode (CPE) using the bacterial strains for the detection of Cr(VI) and Cr(III) ions present in aqueous solutions. (vi) To determine the capability of the developed sensor in the detection of Cr ions in mine water samples collected from Sukinda chromite mine in Odisha, India. (vii) To elucidate the mechanisms occurring at the bio-modified electrode–solution interface. A compendious description of the findings from the present work is given below: The capability of two bacterial strains procured from a microbial culture collection bank (MTCC), Corynebacterium paurometabolum (Gram positive bacterium) and Citrobacter freundii (Gram negative bacterium) as biosorbents for Cr (VI) and Cr(III) ions was assessed. Further, it became of interest to translate the studies related to bioremediation to an industrial situation. For this, bacterial strains were isolated from chromium contaminated water samples collected from surface water of Sukinda chromite mine in Odisha, India. Based on detailed microbiological and molecular biological protocols, two strains of bacteria were identified and characterised as Chromobacterium sp. and Sphingopyxis sp. The bioremediation efficiency of the strains was evaluated taking into consideration the various factors such as effect of contact time of bacterial cells with the chromium ions, pH of the chromium ion solution, biomass loading and initial chromium ion concentration. The Cr(VI) biosorption efficiency obtained for C. freundii was found to be about 59 %, followed by Sphingopyxis sp. and C. paurometabolum ≈ Chromobacterium sp. in the range of 50 % to 55 %. Subsequent to interaction of the bacterial cells with the Cr(VI) solution, the residual chromium was found to be in the form of Cr(III) ions. Hence, complete bioremediation of Cr(VI) could be achieved in terms of both biosorption and bioreduction processes using all the bacterial strains. It was found that the bioreduction process occurring in conjunction with the biosorption process resulted in nil concentration of Cr(VI) ions in the bulk solution. Similarly, studies related to bioremediation of Cr(III) using C. paurometabolum and C. freundii bacterial strains were also performed with higher biosorption efficiency achieved for the former, 50 % compared to 30 % obtained for C. freundii bacterial cells. The bioremediation of Cr(III) ions by the bacterial cells is achieved by the biosorption process. Biosorption of Cr ions by all the bacterial strains were found to follow a typical Langmuirian behaviour. The bioremediation process by the bacterial strains was also evaluated using suitable kinetic models and the results indicated that the bioremediation of Cr(VI) and Cr(III) by C. paurometabolum and C. freundii respectively followed pseudo first order kinetics, while the bioremediation of Cr(VI) by C. freundii, Chromobacterium sp. and Sphingopyxis sp. followed pseudo second order kinetics. It becomes of importance to ascertain the mechanisms of bioremediation of chromium ions by the bacterial cells and for this, different characterisation methods were adopted that helped in deducing the reactions occurring at the bacterial cell surface-chromium solution interface. The involvement of chemical forces in the bioremediation process was corroborated by the achievement of only partial desorption of chromium ions from the biosorbed bacterial cells. This was further confirmed by the Gibbs free energy (∆G) values, which were found to be in the range of -25 to -30 kJ/mol. FTIR spectral studies provided evidence in support of the key functional groups present on the bacterial cell surface such as, –OH, -COOH and –NH, which facilitated the binding with chromium. The EDS data for chromium biosorbed bacterial cells showed peaks corresponding to chromium, thereby confirming the binding of chromium by the bacterial cells. The redox state of chromium bound on the bacterial cell surface was determined with the help of XPS analysis. In the Cr2p XPS spectra obtained for the bacterial cells interacted with Cr(VI), it was interesting to observe a peak corresponding to Cr(III) in addition to Cr(VI), unequivocally indicating that the Cr(III) formed via bioreduction was not only released into the bulk solution but also got biosorbed on the bacterial cell surface. Apparent shifts in the binding energy values for the bacterial cells interacted with chromium were observed in the spectra recorded corresponding to C1s, O1s, N1s, P2p and S2p as compared to the spectra obtained for the bacterial cells alone. This attests to the fact that the functional groups corresponding to the elements mentioned are involved in chemical interaction with the chromium ions or are involved in the donation of electrons to bring about reduction of Cr(VI) to the less toxic Cr(III). The variation in the charge of the bacterial cell surface before and after interaction with chromium ions was monitored by performing zeta potential measurements as a function of pH. The surface charge of the bacterial cells alone was found to be negative over a wide range of pH. Subsequent to interaction of the bacterial cells with the negatively charged oxyanions of Cr(VI) ions, the surface charge was observed to be less electronegative, which further confirmed the binding of the positively charged Cr(III) ions formed via bioreduction on the bacterial cell surface. Similar results were also observed in the case when cells were allowed to interact with Cr(III) ions. The shifts in the iso-electric point for bacterial cells interacted with chromium ions further testified to the involvement of chemical binding forces in the bioremediation process. The findings obtained from the different characterisation methods enabled in understanding the reactions that are occurring at the bacterial cell surface-Cr solution interface. Initially, biosorption via electrostatic interaction of negatively charged oxyanions of Cr(VI) with the positively charged amino groups present on the bacterial cell surface takes place. Subsequent to the biosorption of Cr(VI) ions, the adjacent electron donating functional groups containing ligands present on the bacterial cell surface reduce Cr(VI) to Cr(III) via the reactions shown below: Bioreduction involving –OH group Bioreduction involving –SH group It can be seen that, the reactions involving bioreduction of Cr(VI) in the form of chromate oxyanion to Cr(III) involving hydroxyl and thiol group present on the bacterial cell surface result in the formation of intermediates, chromate-oxy and chromate-thio ester respectively. These intermediates facilitate the transfer of electrons from oxygen/sulphur donor centers to Cr(VI) acceptor molecule, thereby resulting in the reduction of Cr(VI) to Cr(III). The Cr(III) ions thus formed are then either released into the bulk solution or get complexed with the binding groups present on the bacterial cell surface. The next objective was to explore the potential of bacterial strains as sensors for the detection of Cr(VI) and Cr(III) ions. The chromium ions were detected using CV and CSV, both of which are electroanalytical techniques. For this, CPE was coated with the bacterial strains, C. paurometabolum, C. freundii, Chromobacterium sp. and Sphingopyxis sp., and the modified electrode was used as the working electrode in a typical three electrode electrochemical cell. These biosensors developed using each of the aforementioned strains resulted in a ~ 2 to 2.5 fold improved performance compared to the bare CPE for the detection of Cr(VI) ions, due to the binding ability of the various functional groups present on the bacterial cell surface. The lower limit of detection (LLOD) obtained for Cr(VI) and Cr(III) ions using CV technique was found to be 1x10-4 M and 5x10-4 M respectively. The LLOD was further improved to 1x10-9 M and 1x10-7 M for Cr(VI) and Cr(III) respectively using CSV. From the voltammograms obtained, it was postulated that the different functional groups present on the bacterial cell surface facilitate the detection of the chromium ions. Additionally, the developed microbial sensors were also found to be capable of detecting Cr(VI) ions in mine water samples collected from Sukinda chromite mine Odisha, India. In summary, the mechanisms of bioremediation of toxic Cr(VI) ions have been delineated as comprising of both biosorption and bioreduction processes. The residual Cr(VI) concentration subsequent to the treatment of the Cr(VI) aqueous solution with the bacterial cells was found to be nil, which meets the regulatory limit of 0.05 mg L-1 put forward by the US-Environmental Protection Agency (EPA) for a safe effluent discharge. Moreover, it has also been demonstrated that the chosen bacterial strains could be used as sensors for the detection of upto nanomolar concentration of Cr(VI) ions, under optimum conditions.

Pertenece a

ETD at Indian Institute of Science  

Autor(es)

Prabhakaran, Divyasree C - 

Id.: 71376342

Idioma: inglés (Estados Unidos)  - 

Versión: 1.0

Estado: Final

Palabras claveChromium Pollution and Toxicity - 

Tipo de recurso: Thesis  - 

Tipo de Interactividad: Expositivo

Nivel de Interactividad: muy bajo

Audiencia: Estudiante  -  Profesor  -  Autor  - 

Estructura: Atomic

Coste: no

Copyright: sí

Requerimientos técnicos:  Browser: Any - 

Relación: [References] G27310

Fecha de contribución: 06-jun-2018

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