Research Inventy : International Journal of Engineering and Science is published by the group of young academic and industrial researchers with 12 Issues per year. It is an online as well as print version open access journal that provides rapid publication (monthly) of articles in all areas of the subject such as: civil, mechanical, chemical, electronic and computer engineering as well as production and information technology. The Journal welcomes the submission of manuscripts that meet the general criteria of significance and scientific excellence. Papers will be published by rapid process within 20 days after acceptance and peer review process takes only 7 days. All articles published in Research Inventy will be peer-reviewed.
4. optimization of culture condition for enhanced decolorization of reactive ...Darshan Rudakiya
Many synthetic azo dyes and their metabolites are toxic, carcinogenic, and
mutagenic so removal of azo dyes using cost-effective and eco-friendly method is
major aspect.Comamonas acidovorans MTCC 3364 has been routinely reported for
different steroid bioconversion and heavy metal removal. The main purpose of this
study is to check the decolorization efficiency of Comamonas acidovorans MTCC
3364 for different dyes and to optimize the condition which gives maximum
decolorization of Reactive Orange 16 dye. The effect of various physicochemical
parameters including condition, carbon and nitrogen sources, temperature,pH and
dye concentration were studied. The % decolorization of dye was determined by
UV Visible spectroscopy. This bacterial strain efficiently decolorizes Reactive
Orange 16 at 37oC, pH 6.85 within 24 hours giving 99.03 ± 0.5 % dye
decolorization under optimum environmental conditions.
1) Phenolic disinfectants like phenol, 2,4-dichlorophenol, and p-tert-amylphenol bound to Micrococcus lysodeikticus cells, with higher percentages binding to cells for more potent disinfectants.
2) Protoplasts bound slightly less (around 20%) of the phenolic disinfectants compared to whole cells, suggesting cell walls contribute to binding.
3) Binding of 2,4-dichlorophenol decreased with increasing pH, while binding of phenol and p-tert-amylphenol was constant over the pH range tested, relating to differences in ionization properties.
This document provides a summary of various laboratory methods for assessing water quality, including tests for inorganic constituents, hardness, arsenic, alkalinity, pH, biological contaminants, and filtration processes. The methods described include tests for fluoride, iron, sulfate, nitrate, and total dissolved solids. Procedures are outlined for assessing arsenic, alkalinity through titration, and testing for total coliform and E. coli bacteria using UV light and growth indicators. The final section briefly describes a water treatment filter house. The document serves as a training aid for learning water quality analysis techniques.
Most probable number or multiple tube fermentation techniqueSamsuDeen12
multiple tube fermentation or most probable number is a microbiological technique used to check the portability of water. microbial analysis of water is determined, and distinguished between faecal and non faecal contaminated water.
1. Pseudomonas fluorescens was isolated from soil and found to produce an antibacterial substance.
2. The substance was extracted from the bacteria using methylene chloride and purified using silica gel chromatography.
3. Testing showed the substance was stable between pH 5-9 at 37°C for 24 hours, and was soluble in certain solvents but insoluble in others.
Oxidase Test Microbiology - Principle, Procedure, Limitations, Results, QC - in lab #Oxidase Test
As the channel name suggests, our channel will be a perfect lounge for the malayali medicos..we wil be covering videos which will be like lecture classes related to the subjects biochemistry and microbiology in which we are specialised.. It will be a better learning experience for the students especially for those who are not able to understand and follow the normal classes in college..we assure the students that you will get a basic idea regarding the topic and extra reading can be done from the reference textbooks...
If you like my video
#like
#comment
#subscribe my channel
don't forget to subscribe my channel
Qualification
Maneesha M Joseph
MSc MLT (Microbiology)
Assistant Professor
Baby memorial college of allied Health science
Kozhikode
Our Partner Channel
Health & Voyage channel link - https://youtu.be/nzKqRVjlwc0
#Oxidase Test
#Medical
#Microbiology
# malayalam lecturer
#Mallu Medicos Lounge
#MalluMedicosLounge
#MLT
Effect of Diode Laser (805) nm on alpha-toxinsin74
This document summarizes a study that investigated the effect of diode laser (805 nm) irradiation on alpha-toxin production and antibiotic sensitivity in Staphylococcus aureus. Ten S. aureus isolates collected from patient skin samples were exposed to laser light at different time periods. Results showed that alpha-toxin production decreased after 1, 3, and 5 minutes of laser exposure in a time-dependent manner. Laser exposure also slightly increased the antibiotic sensitivity of the S. aureus isolates. The study suggests that low-level laser irradiation may reduce virulence factors and increase antibiotic effectiveness against S. aureus.
This document summarizes an experiment on using the aquatic fern Salvinia molesta to decolorize and detoxify effluent from the textile industry. Key findings include:
- S. molesta was able to decolorize various textile dyes by 48-76% within 60 hours when tested individually, and decolorized Rubine GFL by 97% within 72 hours.
- When exposed to a mixture of dyes and real textile effluent in the lab, S. molesta reduced several pollution parameters like COD and BOD by 65-84% after treatment.
- Anatomical analysis showed changes in the stem of S. molesta when exposed
4. optimization of culture condition for enhanced decolorization of reactive ...Darshan Rudakiya
Many synthetic azo dyes and their metabolites are toxic, carcinogenic, and
mutagenic so removal of azo dyes using cost-effective and eco-friendly method is
major aspect.Comamonas acidovorans MTCC 3364 has been routinely reported for
different steroid bioconversion and heavy metal removal. The main purpose of this
study is to check the decolorization efficiency of Comamonas acidovorans MTCC
3364 for different dyes and to optimize the condition which gives maximum
decolorization of Reactive Orange 16 dye. The effect of various physicochemical
parameters including condition, carbon and nitrogen sources, temperature,pH and
dye concentration were studied. The % decolorization of dye was determined by
UV Visible spectroscopy. This bacterial strain efficiently decolorizes Reactive
Orange 16 at 37oC, pH 6.85 within 24 hours giving 99.03 ± 0.5 % dye
decolorization under optimum environmental conditions.
1) Phenolic disinfectants like phenol, 2,4-dichlorophenol, and p-tert-amylphenol bound to Micrococcus lysodeikticus cells, with higher percentages binding to cells for more potent disinfectants.
2) Protoplasts bound slightly less (around 20%) of the phenolic disinfectants compared to whole cells, suggesting cell walls contribute to binding.
3) Binding of 2,4-dichlorophenol decreased with increasing pH, while binding of phenol and p-tert-amylphenol was constant over the pH range tested, relating to differences in ionization properties.
This document provides a summary of various laboratory methods for assessing water quality, including tests for inorganic constituents, hardness, arsenic, alkalinity, pH, biological contaminants, and filtration processes. The methods described include tests for fluoride, iron, sulfate, nitrate, and total dissolved solids. Procedures are outlined for assessing arsenic, alkalinity through titration, and testing for total coliform and E. coli bacteria using UV light and growth indicators. The final section briefly describes a water treatment filter house. The document serves as a training aid for learning water quality analysis techniques.
Most probable number or multiple tube fermentation techniqueSamsuDeen12
multiple tube fermentation or most probable number is a microbiological technique used to check the portability of water. microbial analysis of water is determined, and distinguished between faecal and non faecal contaminated water.
1. Pseudomonas fluorescens was isolated from soil and found to produce an antibacterial substance.
2. The substance was extracted from the bacteria using methylene chloride and purified using silica gel chromatography.
3. Testing showed the substance was stable between pH 5-9 at 37°C for 24 hours, and was soluble in certain solvents but insoluble in others.
Oxidase Test Microbiology - Principle, Procedure, Limitations, Results, QC - in lab #Oxidase Test
As the channel name suggests, our channel will be a perfect lounge for the malayali medicos..we wil be covering videos which will be like lecture classes related to the subjects biochemistry and microbiology in which we are specialised.. It will be a better learning experience for the students especially for those who are not able to understand and follow the normal classes in college..we assure the students that you will get a basic idea regarding the topic and extra reading can be done from the reference textbooks...
If you like my video
#like
#comment
#subscribe my channel
don't forget to subscribe my channel
Qualification
Maneesha M Joseph
MSc MLT (Microbiology)
Assistant Professor
Baby memorial college of allied Health science
Kozhikode
Our Partner Channel
Health & Voyage channel link - https://youtu.be/nzKqRVjlwc0
#Oxidase Test
#Medical
#Microbiology
# malayalam lecturer
#Mallu Medicos Lounge
#MalluMedicosLounge
#MLT
Effect of Diode Laser (805) nm on alpha-toxinsin74
This document summarizes a study that investigated the effect of diode laser (805 nm) irradiation on alpha-toxin production and antibiotic sensitivity in Staphylococcus aureus. Ten S. aureus isolates collected from patient skin samples were exposed to laser light at different time periods. Results showed that alpha-toxin production decreased after 1, 3, and 5 minutes of laser exposure in a time-dependent manner. Laser exposure also slightly increased the antibiotic sensitivity of the S. aureus isolates. The study suggests that low-level laser irradiation may reduce virulence factors and increase antibiotic effectiveness against S. aureus.
This document summarizes an experiment on using the aquatic fern Salvinia molesta to decolorize and detoxify effluent from the textile industry. Key findings include:
- S. molesta was able to decolorize various textile dyes by 48-76% within 60 hours when tested individually, and decolorized Rubine GFL by 97% within 72 hours.
- When exposed to a mixture of dyes and real textile effluent in the lab, S. molesta reduced several pollution parameters like COD and BOD by 65-84% after treatment.
- Anatomical analysis showed changes in the stem of S. molesta when exposed
The document describes procedures for determining several water quality parameters through laboratory experiments. It discusses determining pH, hardness, turbidity, biochemical oxygen demand (BOD), chemical oxygen demand (COD), carbon dioxide, and alkalinity. For each parameter, it provides an overview, procedure, required apparatus and reagents, and technical discussion of the results and their significance for water quality.
Biochemical test for identification of Bacteria someshwar mankar
This document describes the citrate utilization test procedure and principles. The citrate utilization test is used to determine if an organism can use sodium citrate as its sole carbon source. The test medium contains sodium citrate as the only carbon source and ammonium salts as the sole nitrogen source. If the organism can metabolize the citrate, it will alkalinize the medium which is indicated by a color change of the bromothymol blue indicator from green to blue. Positive results are demonstrated by a color change to deep blue within 24-48 hours.
Disinfection of E.coli using Photocatalytic Sterilization of TiO2 and SiO2 f...Thiwa Wadprom
Subject : Chemical Engineering Project.
bachelor degree of chemical engineering
This project aims to study the efficiency of Eliminate Escherichia coli (E.Coli) bacteria by using the Photocatalytic Sterilization of titanium dioxide (TiO2) and silicon dioxide (SiO2) with different proportions In this project, the experiments were divided into 2 experiments, which were the experiment using the Degradation of Methylene blue method and the E.Coli eradication experiment using culture medium. solid culture and 3M Petrifilm E.Coli count plates to measure results and also invent a prototype (Prototype).
Biochemical tests and physiological tests for various groups of BacteriaHARISH J
This document discusses various biochemical and physiological tests that can be used to identify and characterize different groups of bacteria. It provides details on 14 common biochemical tests including IMVIC, citrate utilization, catalase production, urease, carbohydrate fermentation, and gelatin liquefaction. It also describes 6 physiological tests to determine factors like temperature range, pH tolerance, carbon source utilization, and oxygen relationship. The tests examine the enzymatic activities and metabolic pathways of bacteria, which can help identify them at the genus and species level.
The Rideal-Walker test was developed in 1903 to determine the germicidal value or bactericidal activity of disinfectants. It measures the value of an agent to disinfect cultures of bacteria, with phenol assigned a value of 1 as the standard. The test involves exposing dilutions of the test disinfectant and a phenol control to bacteria cultures and observing bacterial growth to calculate the disinfectant's coefficient relative to phenol. Several countries have since modified the test method but the underlying principle of determining a disinfectant's phenol coefficient remains the same.
This thesis report examines antibiotic resistance due to spices like garlic and turmeric. The objectives were to extract the spices using solvents, test their antimicrobial properties, and determine the minimum inhibitory concentration. Garlic, turmeric, and a combination were extracted using distilled water, ethanol, and chloroform. Extracts showed antibacterial effects against E. coli and Staphylococcus aureus, with ethanol and distilled water being most effective. The minimum inhibitory concentration of extracts was lower than standard antibiotics, indicating they may help reduce antibiotic resistance.
Biochemical tests are based on reactions that takes place in various living rganisms. In microbiology these are useful for identification of various microorganisms like identification and differentiation of various bacterial species. IMViC test is a group of test that are used to differentiate between Escheritia and Enterobacter species.
The document describes the oxidase test, which is used to identify bacteria. It works by detecting cytochrome c, an iron-containing protein in the respiratory chain, which causes an oxidase reagent to turn purple in oxidase-positive bacteria within 5-10 seconds. The document outlines several methods for performing the test, such as using filter paper or commercial strips soaked in reagents like tetramethyl-p-phenylenediamine. It lists both oxidase-positive and -negative bacteria and notes the test is useful for bacterial classification. Modifications to the standard method are presented to more rapidly and accurately detect weakly oxidase-positive organisms.
This document describes an experiment to determine the presence or absence of various secondary metabolites in a plant sample. Nine tests were performed to detect phytochemicals like saponins, glycosides, flavonoids, terpenoids, steroids, carotenoids, alkaloids, anthraquinones, and coumarins. Saponins and coumarins were found to be present, while the other phytochemicals were absent based on the observations from the chemical tests. The experiment was aimed at identifying phytochemicals in a given plant and determining which were present.
Redox Mediated Decolorization and Detoxification of Direct Blue 80 by Partial...Agriculture Journal IJOEAR
Abstract— Textile industries are releasing a large number of toxic synthetic dyes into waste waters. Hence, the removal of such compounds from environment prior to their final disposal is necessary. In the present study, potential use of ginger (Zingiber officinale) peroxidase in decolorization and detoxification of direct blue 80 has been investigated. It was found that only 0.166 U/ml of ginger peroxidase was sufficient for maximum decolorization of dye (25 mg/L). H2O2 was required in low concentration (0.3 mM) in the presence of 0.6 mM 1-hydroxybenzotriazole. Direct blue 80 was also successfully removed in stirred batch process. It was observed that ginger peroxidase was highly stable over a wide range of pH and temperatures. Km and Vmax of the enzyme for direct blue 80 was found to be 27.8 mg/L and 2.09 mg/L/min, respectively. In UV-visible spectral analysis a sharp decline in peak was observed for the treated direct blue 80 which substantiates the breakdown of chromophore group of dye. Genotoxicity assessment by comet assay and chromosomal aberration test confirmed that the direct blue 80 was successfully detoxified by ginger peroxidase. Other direct and acid dyes were also treated either as a single or a mixture of different dyes and it was observed that these dyes were also decolorized significantly under similar experimental conditions. Our study suggests that this enzyme-redox mediator system constitutes a cost effective model which can decolorize the industrial textile effluents and also can reduce the toxic load of environment.
This document summarizes several common biochemical tests used to identify bacteria, including the indole, methyl red (MR), Voges-Proskauer, citrate, and triple sugar iron (TSI) tests. It describes the principle, materials, procedure, and interpretation for each test. The indole test detects the production of indole from tryptophan, while the MR test detects acid production. The Voges-Proskauer test detects acetoin production. The citrate test detects the ability to use citrate as a carbon source. The TSI test detects glucose, lactose and sucrose fermentation and hydrogen sulfide production.
The document discusses methods for identifying bacteria, including phenotypic, immunological, and genetic techniques. Phenotypic methods examine bacterial morphology, staining characteristics, and biochemical reactions. Tests like Gram staining, colony morphology, and catalase can provide initial identification. Further tests of carbohydrate use, enzyme production, and sensitivity to inhibitors allow identification to the species level. Immunological methods detect bacterial antigens, while genetic techniques like PCR and nucleic acid analysis provide accurate identification by examining microbial DNA. Both traditional and molecular methods are used to fully characterize unknown bacteria.
The nitrate reductase test is used to differentiate bacteria based on their ability to reduce nitrate. It detects the intermediate product nitrite, which reacts with reagents to form a red compound. Nitrate-positive bacteria will reduce nitrate to nitrite, showing a color change. If nitrite is not detected, zinc powder is added to catalyze the reduction of any remaining nitrate to nitrite. If a color change then occurs, the bacteria can reduce nitrate but further reduce nitrite; if no color change, the bacteria cannot reduce nitrate. The test helps identify species in the Enterobacteriaceae family and differentiate Neisseria and Corynebacterium species.
This document provides an introduction and overview for a training on wastewater management and protecting drinking water sources. The objectives of the training are outlined, including understanding threats from wastewater like septic systems, wastewater treatment options, project planning activities, and attracting stakeholders. The training overview lists topics that will be covered such as wastewater treatment, onsite wastewater systems, advanced treatment options, and characterization of problems. Additional resources are also listed to supplement the training. Tips are provided for effective training delivery, including knowing the audience, using examples and exercises, and making the content locally relevant. Potential audience types for the training are also outlined.
High organic loaded wastes and wastewater creates environmental pollution. The textile wastewater is a serious environmental challenge faced by Bangladesh textile sub sector. The textile wastewater contains organic compounds and synthetic chemicals. Certain chemicals which are used in the textile industry cause environmental or health problems. Due to presence of chemicals, it cause instance allergic skin reaction or even cause cancer. Environmental degradation by the industrial sector is a matter of serious concern in Bangladesh. Textile processing industries is one of the most important industries of Bangladesh with regard to production, export and labor force employment. The major raw material for textile processing is grey fabric and most of materials consist of cotton and blended fabric. Textile processing employs a variety of chemicals depending on the nature of raw material and products such as; enzymes, detergents, dyes, acids, soda and salt. The textile wastewater is a highly polluted in terms of organic matter and suspended matter such as fibers, grease and chemicals. The textile wastewater is usually hot and alkaline with strong smell and color due to use of chemicals dyes. Studies reveled that wastewater from textile industry is highly toxic and has inhibitory effects on an activated sludge and nitrification.
The document proposes an eco-industrial park at the Port of San Francisco that would include a biodiesel plant and biosolids treatment facilities. The biodiesel plant would be an anchor tenant using feedstock from a rendering plant and water treatment plant. It would supply biodiesel to city and port fleets. The biosolids treatment would move pelletized biosolids by barge instead of trucks to reduce emissions. Adding food waste digesters could produce compost and methane. The eco-industrial park would create jobs and benefits for the community through reduced traffic, noise, pollution and odors.
The document presents a concept for the creation of an industrial park called "CENTRAL" in Kremenchuk, Ukraine. The initiator of the project is the Kremenchuk City Council. The industrial park aims to attract investment, introduce modern technologies, and become a leader in industrial production in the region. It will have five sectors focusing on various industries like engineering, logistics, IT, and alternative energy. The 168.55 hectare park is expected to generate over $422 million USD in investment and create over 3,500 new jobs. It will improve the local economy and standards of living.
This document discusses treatment of wastewater from a water jet loom machine in the textile industry. It compares the efficiency of chemical coagulation and electrocoagulation methods. For chemical coagulation, the type and amount of coagulant and coagulant aids, pH, and stirring rate significantly impacted treatment efficiency. The optimum conditions removed 89% of turbidity, 85% of COD, and 71% of oil. For electrocoagulation, electrode material, electric potential, and contact time were significant. The optimum electrocoagulation conditions removed 99% of turbidity, 97% of COD, and 87% of oil.
The document describes procedures for determining several water quality parameters through laboratory experiments. It discusses determining pH, hardness, turbidity, biochemical oxygen demand (BOD), chemical oxygen demand (COD), carbon dioxide, and alkalinity. For each parameter, it provides an overview, procedure, required apparatus and reagents, and technical discussion of the results and their significance for water quality.
Biochemical test for identification of Bacteria someshwar mankar
This document describes the citrate utilization test procedure and principles. The citrate utilization test is used to determine if an organism can use sodium citrate as its sole carbon source. The test medium contains sodium citrate as the only carbon source and ammonium salts as the sole nitrogen source. If the organism can metabolize the citrate, it will alkalinize the medium which is indicated by a color change of the bromothymol blue indicator from green to blue. Positive results are demonstrated by a color change to deep blue within 24-48 hours.
Disinfection of E.coli using Photocatalytic Sterilization of TiO2 and SiO2 f...Thiwa Wadprom
Subject : Chemical Engineering Project.
bachelor degree of chemical engineering
This project aims to study the efficiency of Eliminate Escherichia coli (E.Coli) bacteria by using the Photocatalytic Sterilization of titanium dioxide (TiO2) and silicon dioxide (SiO2) with different proportions In this project, the experiments were divided into 2 experiments, which were the experiment using the Degradation of Methylene blue method and the E.Coli eradication experiment using culture medium. solid culture and 3M Petrifilm E.Coli count plates to measure results and also invent a prototype (Prototype).
Biochemical tests and physiological tests for various groups of BacteriaHARISH J
This document discusses various biochemical and physiological tests that can be used to identify and characterize different groups of bacteria. It provides details on 14 common biochemical tests including IMVIC, citrate utilization, catalase production, urease, carbohydrate fermentation, and gelatin liquefaction. It also describes 6 physiological tests to determine factors like temperature range, pH tolerance, carbon source utilization, and oxygen relationship. The tests examine the enzymatic activities and metabolic pathways of bacteria, which can help identify them at the genus and species level.
The Rideal-Walker test was developed in 1903 to determine the germicidal value or bactericidal activity of disinfectants. It measures the value of an agent to disinfect cultures of bacteria, with phenol assigned a value of 1 as the standard. The test involves exposing dilutions of the test disinfectant and a phenol control to bacteria cultures and observing bacterial growth to calculate the disinfectant's coefficient relative to phenol. Several countries have since modified the test method but the underlying principle of determining a disinfectant's phenol coefficient remains the same.
This thesis report examines antibiotic resistance due to spices like garlic and turmeric. The objectives were to extract the spices using solvents, test their antimicrobial properties, and determine the minimum inhibitory concentration. Garlic, turmeric, and a combination were extracted using distilled water, ethanol, and chloroform. Extracts showed antibacterial effects against E. coli and Staphylococcus aureus, with ethanol and distilled water being most effective. The minimum inhibitory concentration of extracts was lower than standard antibiotics, indicating they may help reduce antibiotic resistance.
Biochemical tests are based on reactions that takes place in various living rganisms. In microbiology these are useful for identification of various microorganisms like identification and differentiation of various bacterial species. IMViC test is a group of test that are used to differentiate between Escheritia and Enterobacter species.
The document describes the oxidase test, which is used to identify bacteria. It works by detecting cytochrome c, an iron-containing protein in the respiratory chain, which causes an oxidase reagent to turn purple in oxidase-positive bacteria within 5-10 seconds. The document outlines several methods for performing the test, such as using filter paper or commercial strips soaked in reagents like tetramethyl-p-phenylenediamine. It lists both oxidase-positive and -negative bacteria and notes the test is useful for bacterial classification. Modifications to the standard method are presented to more rapidly and accurately detect weakly oxidase-positive organisms.
This document describes an experiment to determine the presence or absence of various secondary metabolites in a plant sample. Nine tests were performed to detect phytochemicals like saponins, glycosides, flavonoids, terpenoids, steroids, carotenoids, alkaloids, anthraquinones, and coumarins. Saponins and coumarins were found to be present, while the other phytochemicals were absent based on the observations from the chemical tests. The experiment was aimed at identifying phytochemicals in a given plant and determining which were present.
Redox Mediated Decolorization and Detoxification of Direct Blue 80 by Partial...Agriculture Journal IJOEAR
Abstract— Textile industries are releasing a large number of toxic synthetic dyes into waste waters. Hence, the removal of such compounds from environment prior to their final disposal is necessary. In the present study, potential use of ginger (Zingiber officinale) peroxidase in decolorization and detoxification of direct blue 80 has been investigated. It was found that only 0.166 U/ml of ginger peroxidase was sufficient for maximum decolorization of dye (25 mg/L). H2O2 was required in low concentration (0.3 mM) in the presence of 0.6 mM 1-hydroxybenzotriazole. Direct blue 80 was also successfully removed in stirred batch process. It was observed that ginger peroxidase was highly stable over a wide range of pH and temperatures. Km and Vmax of the enzyme for direct blue 80 was found to be 27.8 mg/L and 2.09 mg/L/min, respectively. In UV-visible spectral analysis a sharp decline in peak was observed for the treated direct blue 80 which substantiates the breakdown of chromophore group of dye. Genotoxicity assessment by comet assay and chromosomal aberration test confirmed that the direct blue 80 was successfully detoxified by ginger peroxidase. Other direct and acid dyes were also treated either as a single or a mixture of different dyes and it was observed that these dyes were also decolorized significantly under similar experimental conditions. Our study suggests that this enzyme-redox mediator system constitutes a cost effective model which can decolorize the industrial textile effluents and also can reduce the toxic load of environment.
This document summarizes several common biochemical tests used to identify bacteria, including the indole, methyl red (MR), Voges-Proskauer, citrate, and triple sugar iron (TSI) tests. It describes the principle, materials, procedure, and interpretation for each test. The indole test detects the production of indole from tryptophan, while the MR test detects acid production. The Voges-Proskauer test detects acetoin production. The citrate test detects the ability to use citrate as a carbon source. The TSI test detects glucose, lactose and sucrose fermentation and hydrogen sulfide production.
The document discusses methods for identifying bacteria, including phenotypic, immunological, and genetic techniques. Phenotypic methods examine bacterial morphology, staining characteristics, and biochemical reactions. Tests like Gram staining, colony morphology, and catalase can provide initial identification. Further tests of carbohydrate use, enzyme production, and sensitivity to inhibitors allow identification to the species level. Immunological methods detect bacterial antigens, while genetic techniques like PCR and nucleic acid analysis provide accurate identification by examining microbial DNA. Both traditional and molecular methods are used to fully characterize unknown bacteria.
The nitrate reductase test is used to differentiate bacteria based on their ability to reduce nitrate. It detects the intermediate product nitrite, which reacts with reagents to form a red compound. Nitrate-positive bacteria will reduce nitrate to nitrite, showing a color change. If nitrite is not detected, zinc powder is added to catalyze the reduction of any remaining nitrate to nitrite. If a color change then occurs, the bacteria can reduce nitrate but further reduce nitrite; if no color change, the bacteria cannot reduce nitrate. The test helps identify species in the Enterobacteriaceae family and differentiate Neisseria and Corynebacterium species.
This document provides an introduction and overview for a training on wastewater management and protecting drinking water sources. The objectives of the training are outlined, including understanding threats from wastewater like septic systems, wastewater treatment options, project planning activities, and attracting stakeholders. The training overview lists topics that will be covered such as wastewater treatment, onsite wastewater systems, advanced treatment options, and characterization of problems. Additional resources are also listed to supplement the training. Tips are provided for effective training delivery, including knowing the audience, using examples and exercises, and making the content locally relevant. Potential audience types for the training are also outlined.
High organic loaded wastes and wastewater creates environmental pollution. The textile wastewater is a serious environmental challenge faced by Bangladesh textile sub sector. The textile wastewater contains organic compounds and synthetic chemicals. Certain chemicals which are used in the textile industry cause environmental or health problems. Due to presence of chemicals, it cause instance allergic skin reaction or even cause cancer. Environmental degradation by the industrial sector is a matter of serious concern in Bangladesh. Textile processing industries is one of the most important industries of Bangladesh with regard to production, export and labor force employment. The major raw material for textile processing is grey fabric and most of materials consist of cotton and blended fabric. Textile processing employs a variety of chemicals depending on the nature of raw material and products such as; enzymes, detergents, dyes, acids, soda and salt. The textile wastewater is a highly polluted in terms of organic matter and suspended matter such as fibers, grease and chemicals. The textile wastewater is usually hot and alkaline with strong smell and color due to use of chemicals dyes. Studies reveled that wastewater from textile industry is highly toxic and has inhibitory effects on an activated sludge and nitrification.
The document proposes an eco-industrial park at the Port of San Francisco that would include a biodiesel plant and biosolids treatment facilities. The biodiesel plant would be an anchor tenant using feedstock from a rendering plant and water treatment plant. It would supply biodiesel to city and port fleets. The biosolids treatment would move pelletized biosolids by barge instead of trucks to reduce emissions. Adding food waste digesters could produce compost and methane. The eco-industrial park would create jobs and benefits for the community through reduced traffic, noise, pollution and odors.
The document presents a concept for the creation of an industrial park called "CENTRAL" in Kremenchuk, Ukraine. The initiator of the project is the Kremenchuk City Council. The industrial park aims to attract investment, introduce modern technologies, and become a leader in industrial production in the region. It will have five sectors focusing on various industries like engineering, logistics, IT, and alternative energy. The 168.55 hectare park is expected to generate over $422 million USD in investment and create over 3,500 new jobs. It will improve the local economy and standards of living.
This document discusses treatment of wastewater from a water jet loom machine in the textile industry. It compares the efficiency of chemical coagulation and electrocoagulation methods. For chemical coagulation, the type and amount of coagulant and coagulant aids, pH, and stirring rate significantly impacted treatment efficiency. The optimum conditions removed 89% of turbidity, 85% of COD, and 71% of oil. For electrocoagulation, electrode material, electric potential, and contact time were significant. The optimum electrocoagulation conditions removed 99% of turbidity, 97% of COD, and 87% of oil.
1) Anaerobic treatment is a biological process that occurs without oxygen to stabilize organic materials by converting them to methane, carbon dioxide, and ammonia.
2) It has several advantages over aerobic treatment including lower energy requirements, energy generation in the form of methane gas, and lower sludge production.
3) The process involves several groups of microorganisms that break down organic matter in stages through hydrolysis, acidogenesis, acetogenesis, and methanogenesis.
Indian textile industy environmental issues pptAdane Nega
The document discusses the environmental issues faced by the Indian textile industry. It provides details on the structure and size of the industry, which is one of the largest in India. The decentralized sector, which includes powerlooms and handlooms, accounts for the majority of textile production. Wet processing poses serious pollution problems due to the large quantity of chemicals used. Strict environmental regulations have been introduced but enforcement remains a challenge. The industry needs to modernize and adopt cleaner technologies to compete globally in a sustainable manner.
Lecture notes of Environmental Engineering-II as per Solapur university syllabus of TE CIVIL.
Prepared by
Prof S S Jahagirdar,
Associate Professor,
N K Orchid college of Engg and Technology,
Solapur
The document discusses various aspects of anaerobic wastewater treatment processes. It provides information on the types and characteristics of anaerobic reactors including UASB and EGSB reactors. It also describes the formation of anaerobic granular sludge, which allows high biomass retention and efficient COD removal. Additionally, it compares the kinetics, environmental factors, and advantages of anaerobic versus aerobic wastewater treatment processes.
This document discusses the treatment of wastewater from the textile industry. It notes that textile wastewater contains a variety of dyes and chemicals from dyeing and finishing processes that make it challenging to treat. The major pollutants in textile wastewater include high levels of suspended solids, chemical oxygen demand, heat, color, acidity, and other soluble substances. The document then outlines the primary, secondary, and tertiary treatment processes used to remove these pollutants, including screening, sedimentation, neutralization, flocculation, aerated lagoons, activated sludge processes, and various advanced oxidation processes.
The document discusses different types of chemical reactors used in industrial processes. It describes basic reactor components like tanks and pipes and operating modes like batch, continuous stirred-tank, and plug flow reactors. Key aspects covered include material and heat transfer, reaction rates, and the influence of temperature, pressure and catalysts. Common reactor designs are presented, such as jackets, coils and packed beds for heat exchange. The document also discusses homogeneous and heterogeneous catalysis.
The document describes the effluent treatment plant (ETP) at Rourkela Steel Plant. The ETP treats wastewater from the Gas Cleaning Plant and recycles it for further use. The wastewater contains high levels of suspended solids that are removed through a multi-step process involving flash mixing with coagulants, settling in clarifiers, dewatering using a filter press, and recycling of treated water. The ETP is designed to treat 1140 cubic meters of wastewater per hour to reduce costs and conserve water resources.
An Introduction To Wastewater And Sludge PrinciplesOliver Grievson
The document provides an overview of wastewater and sewage sludge treatment processes. It discusses the history of sewage treatment in the UK from the 14th century to present day. It then describes the basic treatment processes including preliminary treatment to remove solids, primary settlement to separate liquids and solids, secondary biological treatment using trickling filters or activated sludge plants, and sludge handling which includes thickening, digestion, and dewatering processes.
Effluent Treatment Plant Design, Operation And Analysis Of Waste Water Jaidev Singh
Contents
1. Introduction to Effluent Treatment Plant (ETP)
1.1 Use of water in industries
1.2 Industrial waste water sources
1.3 Effluent Treatment Plant
1.4 National Standards for waste water
1.5 What do these standards means?
1.6 Waste water treatment
1.7 Planning an Effluent Treatment Plant : Factors to Consider
2. Treatment Methods
2.1 Physical Unit Operations
2.2 Chemical Unit Processes
2.3 Biological Unit Processes
3. Operation and control
3.1 Mixed liquor suspended solids
3.2 Sludge Volume Index and Sludge Density Index
3.3 Sludge Age; Mean Cell Residence Time (MCRT)
3.4 Food/Mass Ratio
3.5 Constant MLSS
3.6 Return Activated Sludge Control (RAS)
4. Choosing an Effluent Treatment Plant
4.1 Biological Treatment
4.2 Physico-Chemical Treatment
4.3 Physico-Chemical and Biological Treatment
4.4 Area Requirement Comparison
4.5 Cost Comparison
5. Chemical Analysis of Waste Water
5.1 Commonly used chemicals
5.2 Chemical Tests and procedures
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Research Inventy : International Journal of Engineering and Science
1. RESEARCH INVENTY: International Journal of Engineering and Science
ISSN: 2278-4721, Vol. 1, Issue 12 (December 2012), PP 54-60
www.researchinventy.com
Biodegradation of Azo Dye Reactive Red BL by Alcaligenes Sp. AA09
1
Ajay Kumar Pandey, 2Vinay Dubey
1, 2,
(Department of Biotechnology, School Of Bioengineering, SRM University, Kattankulatur, Chennai,
Tamilnadu 603203, India)
Abstract- A dye-decolorizing bacterium strain was isolated from activated sludge of a textile printing
wastewater treatment plant of Perundurai. The strain was identified as Alcaligenes sp. AA09 based on Gram
staining and sequence analysis of 16S rDNA gene. Alcaligenes sp. AA09 decolorized more than 90% of the azo
dye reactive red BL within 24 h under aerobic condition at pH 7.0 and temperature 250C with 50-200mg/l dye
concentration. A 60Kd NADH/NADPH-dependent, oxygen sensitive, soluble cytosolic enzyme, azoreductase was
responsible for degradation of reactive red BL azo dye. The bacterial dye degradation was confirmed by
analysis of the degraded products using TLC and GC-MS analysis.
Key Words- Biodegrdation, Reactive Red BL, Aerobic, Decolorization, Azo dyes, Azoreductase, phytotoxicity
I. Introduction
Global industrialization has resulted in the release of large amounts of potentially toxic, mutagenic and
xenobiotic compounds into the biosphere [1-2]. Cleaning up of the environment by the removal of hazardous
contaminants is a crucial and challenging problem needing numerous approaches to reach long-lasting suitable
solutions [3]. The textile industries are using synthetic dyes with ease of production, fastness and variety in
colour compared to natural dyes and daily discharging millions of liters of untreated effluent containing harmful
dye wash into receiving water bodies posing serious health problems [4-6]. An average textile mill produces 60
x 104 m of fabric and discharges approximately 1.5 million liters of effluent per day in India [3-9]. Azo dyes,
with a total annual production 7 x105 metric tons worldwide production are commercially available and are
typically 5-10% of this amount is discharged into environment which is usually recalcitrant to conventional
wastewater treatment methods [10]. Presence of dyes in aqueous ecosystem diminishes the photosynthesis by
impeding the light penetration into deeper layer thereby deteriorating the water quality and lowering the gas
solubility [11-13]. To avoid these problems, the effluent from textile industries must be treated before their
discharge. During the past three decades, several physical, chemical and biological decolorization methods have
been accepted by the paper and textile industries [12-15]. Wide range o microorganisms including bacteria [9],
fungi, yeasts, actinomycetes and algae capable of degrading azo dyes have been reported of them bacterial and
fungal cultures have found to cause decolourization of various azo dyes to a great extent [15-16]. Azoreductase
enzymes responsible for decolourisation of azo dyes are purified from several bacterial strains [16-18].
The present investigation was aimed to exploit the biodegradation abilities of indigenous microbial
flora for remediative purposes. In the present study we screened and isolated bacterial strains organisms for the
ability to decolorize azo dyes present in the textile effluents as well as the commercially available azo dye
reactive red BL. The dye degraded products after the microbial treatment would be analyzed by TLC and GC-
MS. Further the study of enzyme involved in dye degradation and partial purification of the same was carried
out.
II. Materials And Methods
2.1. Dyes and Media
Azo dye Reactive Red BL was provided by Department of textile industry, Anna University, Chennai,
India. All media components and chemicals used in the studies were of analytical grade.
2.2. Sample Collection
The textile dye effluent samples were collected from Common effluent treatment plant, Perundurai in
sterile plastic containers. The samples were brought to the laboratory and processed within 48 h.
2.3. Isolation of bacteria from Textile effluent
Serial dilution of textile dye effluent sample has been done from 10-1 to 10-6 by using distilled water.
One hundred µl of effluent sample was inoculated on LB medium by using sterilized glass rod spreader and
incubated at 25oC for 48 h. After 48 h of incubation morphologically distinct colonies were picked and
transferred in 1 ml LB broth and incubated at 250C for 48h.The pure culture was prepared and stored at 40C for
further studies.
54
2. Biodegradation Of Azo Dye Reactive Red BL By Alcaligenes Sp. AA09
2.4. Decolorization of textile dye effluent by isolated bacterial strains
For checking the dye effluent decolorization by bacterial strains, decolorization assay was performed,
in which the bacterial colonies from pure culture were transferred to 25 ml LB broth and allowed to grow for
48h at 370C. The effluent was mixed with the bacterial culture at 1:1(v/v) and 3:1(v/v) ratio and incubated at
30°C under shaking conditions for 48 hrs. Uninoculated effluent with broth served as control.Decolorization
was determined in 5 ml aliquots from each flask after 48 hrs of incubation. The culture was centrifuged at
10,000 rpm for 15 min to remove the cells. The absorbance of the supernatants was measured at 533 nm using
spectrophotometer at regular intervals of 1h during the decolorization process.
2.5. Enrichment of dye decolorizing bacterial strains
The bacterial strains capable of decolorizing the textile effluent efficiently were enriched by growing
into conical flasks containing 100mL (pH 7.0) of MSM medium (gL-1 ): (Na2HPO4 , 1.264; KH2PO4, 0.326;
NH4Cl, 1.0; MgSO4, 0.098; CaCl2. 2H2O, 0.044; FeSO4.7 H2O, 0.01 and NaCl, 0.1) amended with 50mg/L dye
as the sole source of C and N. The MSM medium containing dye were inoculated with the strains that showed
efficient decolorization and incubated at 250C for 24 hrs under static condition, cell suspension from each flask
were plated onto MSM agar medium and incubated at 300C for 48 hrs. Morphologically distinct colonies were
selected and were purified by streaking twice on LB agar medium. Single colony were transferred on to agar
slant and stored at 40C for further studies [8].
2.6. Dye decolorization assay
The decolorization assay of azo dye, Reactive Red BL was carried out in 100ml MSM medium at 200
mg/L dye concentration with inoculum concentration 10% v/v at 25 °C and 150 rpm. Two types of control were
used, uninoculated sterile medium supplemented with dye (abiotic control) while in other inoculated medium in
which dye has been omitted (biotic control). After incubation at shaking condition to a sampling time 3h aliquot
(2ml), culture media was withdrawn, centrifuged at 10,000 rpm for 10 min to separate the bacterial cell mass
and clear supernatant.Decolorization of dyes was determined by monitoring the decrease in absorbance at the
maximum absorption wavelength (λmax.)at 518 nm for Reactive Red BL in an UV-Visible scanning
spectrophotometer. Decolorization activity was calculated by applying the following formula [2]:
A A
Decolorization (%) = 0 t x 100
A
0
A0- Initial Absorbance
At – Observed Absorbance
2.7. Effect of different concentrations of dye on decolorization
To check the efficiency of dye decolorization by the bacterial isolate in question decolorization assay
was carried out at different concentration of dye Reactive Red BL, 50 mg/L, 100 mg/L, 200 mg/L, 300 mg/L,
400mg/L, 500 mg/L. Samples were withdrawn at the interval of 3h and subjected to centrifugation at 10,000
rpm and supernatant was used to determine the decolorization of dye by UV-Vis spectrophotometer at 518 nm
and the % decolourization was calculated [2].
2.8. Effect of incubation conditions on dye decolorization
In a conical flask 100ml LB broth, 200mg/L dye and inoculum (10% v/v) were added. One set of flasks
were incubated at 25OC under static conditions and the other at same temperature on an orbital shaker at
150rpm. Appropriate controls were used as mentioned above. Samples were withdrawn at regular interval of 3h,
centrifuged at 10,000 rpm for 10 min and the absorbance of supernatant was read at 518 nm in a
spectrophotometer and the % decolorization was calculated.
2.9. Effect of yeast extract as a co substrate
In this experiment dye concentration was gradually increased from 100 mg/L to 500 mg/L while yeast
extract concentration decreased from 400 mg/L. To the flask containing 500 mg/L dye, no yeast extract was
added. Two controls have been used, one is abiotic without inoculation while other with 500mg/lit yeast with
inoculation in which dye has been omitted. The inoculated flasks were incubated at 25 OC in stationary
condition. Concentration of dye has been determined by taking absorbance at 518 nm in regular intervals in a
spectrophometer and decolorization calculated [19-20].
55
3. Biodegradation Of Azo Dye Reactive Red BL By Alcaligenes Sp. AA09
2.10. Effect of pH on decolorization
For determination of pH dependence of bacterial bio-decolorization, medium was supplemented with
Reactive Red BL dye at low concentration of dye 200 mg/lit. The medium pH was adjusted between 5.0 to 9.0
with hydrochloric acid and sodium hydroxide and inoculated with 10% (v/v) bacterial culture and incubated at
25Oc temperature. Samples have been collected from regular time intervals up to 24 hrs. Decolorization % was
calculated based on the OD values at 518 nm [21].
2.11. Identification and characterizationof Dye decolorizing bacterial strain
The dye decolorizing bacterium was found to be a Gram-negative, rod-shaped and motile. For
identification of bacterial strain the genomic DNA was extracted from culture [17-18]. The universal primers
were used and PCR amplification was performed in total reaction volume 20 μL. The PCR programming used
were 94 °C for 5 min, 30 cycles at 94 °C for 1 min, 54 °C for 1 min and 72 °C for 3 min and final extension was
at 72 °C for 10 min. The amplified PCR product was detected on 0.8% agarose gel in TBE buffer. Nucleotide
sequences were determined by DNA sequencing method. The sequences of 16S r DNA gene of the bacterial
strain were subjected to BLAST search tool in gene database of NCBI.
2.12. Azoreductase activity in crude protein extract
The culture was grown in minimal medium containing 25mg l −1 of Reactive Red BL until the dye was
completely decolorized. Cells were harvested by centrifugation, washed with 50 mM phosphate buffer (pH 7)
and resuspended in the same buffer. Cells were then disrupted by sonication (four cycles of 15 s, 75%
amplitude;), and cell debris was removed by centrifugation at 4°C. Supernatant was used as the crude protein
extract. The enzyme assay mixture was assayed as consisted of 50 mM phosphate buffer (pH 7), 2 mM NADH,
20 μM Reactive Red BL, and 0.1 ml of crude protein extract in a total volume of 1 ml. Dye decolorization was
monitored spectrophotometrically at 518 nm. Azoreductase activity in the presence of NADPH was measured in
a similar manner by the addition of 2 and 0.2 mM of NADPH, to the assay mixture. Enzyme activity was
expressed in units of micromoles of dye reduced per minute. Protein content of the crude protein extract was
determined by Lowry’s method using bovine serum albumin as standard for determination of specific activity,
i.e., enzyme units per milligram protein.
2.12. Partial purification of Azoreductase
The enzyme responsible for dye decolorization has been partially purified by a combination of different
purification processes. Crude extract was subjected to ammonium sulphate ((NH4)2SO4) precipitation at 40%
saturation to remove impurities, followed by 60% saturation in a second step to precipitate the azoreductase.
The precipitated proteins from the second precipitation step have been collected by centrifugation, and the pellet
has been dissolved in 30 mL of phosphate buffer (50 mM, pH7.0).The precipitated proteins were desalted by
dialysis against phosphate buffer (50 mM, pH7.0) overnight. The desalted solution after dialysis has been
filtered through a 0.45 μm filter and was subjected to anion exchange chromatography using DEAE cellulose as
column bed in 1.5 cm diameter with bed length 10 cm and applied 1.5 mL aliquots of the resulting solution to
the column. Protein was eluted at 5.4 cm/h. with the sample buffer and with step wise increasing concentration
of buffer containing NaCl with range 0-200 mM and highest concentration up to 1 M. The 1.5 mL fractions
were collected for each concentration of NaCl. The protein concentration and enzyme activity has been
determined at every steps of purification.
2.12. SDS-PAGE
In order to ascertain the presence of the enzyme, SDS PAGE was carried out using the sample.
Resolving gel and Stacking were prepared and poured into the gel apparatus and allowed to polymerize. Sample
buffer (β-merceptoethenol and bromophenol blue) was added into sample with volume 1:5 and the mixture was
heated at 100 °C for 2 min and then spun down for a few seconds and then loaded in the wells of acrylamide
polymerized gel in gel apparatus. The marker was also loaded in separate well to determine the molecular
weight. The electrodes were attached to the gel apparatus and power supply was turned on at 150 V.
Electrophoresis was carried out at a constant voltage until the dye reaches about 1 cm from the end of the gel.
Power was turned off and the gel was taken off with the help of spacer. The gel was submerged in staining
solution and allowed the gel to stain overnight. Gel was then placed in destaining solution to remove
background stain. Protein bands were clearly visible after destaining.
56
4. Biodegradation Of Azo Dye Reactive Red BL By Alcaligenes Sp. AA09
2.13. Extraction and analysis of degraded product
The bacterial strain was inoculated in minimal medium containing 100 mg/l of Reactive Red BL and
incubated under ambient conditions. After complete decolorization, 40 ml of dye degraded samples was taken
and centrifuged at 10000 rpm for 10 min. Then the supernatant was filtered through Whatman #1 filter paper.
Filtrate was then extracted three times with diethyl ether pooled and evaporated the extracted product to dryness
[21].The degraded products were analyzed by TLC and GC MS. For TLC the solvent system butenol: water:
acetic acid = 5:3:2 was poured it into the TLC tank. The extracted sample was dissolved in 0.5 ml of methanol
and was loaded on TLC plate of 3.5 × 5.0 cm and allowed to run in a TLC tank. After the run was complete the
TLC plate was taken out and air dried. The plate was observed under UV light and the Rf values of the bands
were recorded. The extracted metabolites were also analyzed by the Gas chromatography-mass spectrometry
carried out with JEOL GC Mate-II fitted with HP5 column.
III. Results and Discussion
3.1. Screening and Identification
The textile dye effluent sample of common textile effluent treatment plant and textile industries from
Perundurai and Erode area have given morphologically distinct colonies were picked up, showing decolorization
zone on LB agar plates containing Reactive Red BL dye. The isolates were used for textile effluent
decolorization assay and the maximum decolorization showing isolate was used for the decolorization in conical
flasks. The efficiency of decolorization was calculated. The selected bacterial strain 006/a/PP/I* has shown
maximum efficiency in decolorization of the mixture of textile effluent. The flask which contained effluent and
broth in 1:1(v/v) ratio showed more decolorization in comparison to 3:1 (v/v) ratio which was 81.1 % and 46 %
respectively after 24 h of incubation (Fig.1).
Fig.1. Textile Effluent Decolorization assay
The effect of incubation conditions namely shaking and stationary condition on decolorization of
Reactive Red BL by 006/a/PP/I*, revealed that static condition was more suitable for decolorization, where the
activity was found to be 95% and at shaking condition it was 84.83%. . The data suggest that in static condition
is more appropriate for the decolorization of the dye by the bacterium (fig.2).
Figure.2. Effect of Shaking and static condition on dye decolorization
57
5. Biodegradation Of Azo Dye Reactive Red BL By Alcaligenes Sp. AA09
Different concentrations of azo dye Reactive Red BL, ranges from 50,100,200,300,400,500mg/lit has
been taken for the determination of the effect of dye concentration on the decolorization. In 48 hrs it has been
found that with increase in dye concentration the dye decolourizing efficiency of the bacterial strain decreases.
The maximum decolorization was found on 50mg/lit concentration and minimum decolorization was found on
500mg/lit concentration (fig.3).
Fig
ure.3. Effect of dye concentration on decolorization efficiency of Alcaligenes sp. AA09
Different concentrations of yeast extract (100–500 mg/l) have been utilized the determination of effect
of yeast extract as co substrate for decolorization efficiency of bacterial strain (Fig 14). With increase
concentration of azo dye (0-500mg/lit) and decrease concentration of yeast extract (500-0mg/lit) the time taken
for decolorization was found to be 72hrs. At 500mg/L dye concentration and in absence of yeast extract the
decolorization after 70 h was found to be 78.91% suggesting that the organism could utilize azo dye as a sole C-
source (fig.4).
Fig
ure.4. Utilization of Azo Dye Reactive Red BL as Sole C-source
Effect of pH on decolorization indicated that the bacterial culture generally exhibited maximum
decolorization rate at pH values near 7 (Fig 5). An increase in pH from 5.0 to 9.0 did not show any marked
changes in the values of percentage decolorization indicating that the organism could be used in treatment of
effluents which are either acidic or alkaline. The optimum pH for decolorization of azo dye Reactive Red BL
however remains 7.0.
58
6. Biodegradation Of Azo Dye Reactive Red BL By Alcaligenes Sp. AA09
Figure. 5. Effect of pH on the decolorization of azo dye Reactive Red BL
3.2. Identification of dye decolorizing bacterial strain
The dye decolorizing bacterial strain was identified by 16S rDNA gene sequence analysis of 387 base
pair and it was designated as Alcaligenes sp. AA09.
3.3. Determination of azoreductase activity
Crude protein extract obtained from Alcaligenes sp. AA09 cells was found to decolorize Reactive Red
BL dye using NADH as electron donor. The absorbance of the enzyme assay mixture was taken at 518nm by
using spectrophotometer at the 5 min of interval. The absorbance was found to be decrease, which indicates that
the NADH is working as coenzyme for Azoreductase. By using Lowry method the protein concentration was
found to be 180μg/ml in crude extract and the enzyme activity was found to be 0.025 U/mg. Azoreductase
enzyme was visualized from the crude protein extract on a zymogram. The development of zymogram was
based on the decolorization of Reactive Red BL stained gel by azoreductase in presence of NADH, which could
be seen as a clear band against a red background. The enzyme involved in dye decolorization was partially
purified by a combination of purification methods by using ammonium sulphate (70% concentration)
precipitation (0.067 U/mg; 140μg/ml), dialysis (0.072 U/mg; 120μg/mL), anion exchange column
chromatography (0.081 U/mg; 90μg/mL), and gel filtration chromatography (0.086 U/mg; 85μg/mL). The
molecular weight of purified azoreductase was determined by gel electrophoresis using standard marker. The
partially purified azo reductase revealed band on 10% SDS-PAGE with an estimated molecular weight of 60
KDa approximately.
3.4. Analysis of dye degraded products
The TLC analysis of dye degraded products shown Rf value of the dye sample 0.42, and the Rf value
of degraded product was found to be 0.02. Therefore, the TLC analysis has confirmed that there was no starting
material, present in the degraded sample and two new degraded products were formed For confirmation of the
dye degradation Gas chromatography-mass spectrometry was carried out with Jeol GC Mate-II fitted with HP5
column. The dye and degraded samples were injected into a GC/MS (Fig. 24). First the blue dye sample,
handpicked from aquous solution, showed the elution of the dye’s expected mass 984.208 at retention time
1.78 min. Then the degraded sample was injected in normal temperature gradient to find out the residual amount
of compound remains. The GC did not show any peak at 1.78 min. The following GC have shown three major
elution at 10.4 and 20.58 min. The respective mass spectra does not show any mass with 984.208. So, it is
concluded that no starting material, i.e. Reactive Red BL dye, remains in the culture media Despite the fact that
untreated dyeing effluents might cause serious environmental and health hazards, they are being disposed off in
water bodies and this water is being used for an agriculture purpose. Use of untreated and treated dyeing
effluents in the agriculture has direct impact on fertility of soil. Thus, it was of concern to assess the
phytotoxicity of the dye before and after degradation.
59
7. Biodegradation Of Azo Dye Reactive Red BL By Alcaligenes Sp. AA09
IV. Conclusion
Toxic azo dye Reacive Red BL was completely biodegraded by Alcaligenes sp. AA09 bacterial strain,
isolated from textile dye wastewater, it required less incubation time for decolorization under static condition.
Enzymatic studies indicated the involvement of NADH/NADPH-dependent enzyme Azoreductase in the
biotransformation at pH 7.0 and temperature 250C with 50-200mg/l dye concentration, within 24 h, under
aerobic condition. TLC and GC-MS analysis confirmed the complete biodegradation transformation of Reactive
Red BL. The forerunning results suggest the potential of utilizing Alcaligenes sp. AA09 to decolorize textile
effluent containing textile azodye Reactive Red BL via appropriate bioreactor operations. The prospect plan of
work focused on the pilot scale treatment of textile tannery effluent, based on laboratory scale research by using
free and immobilized cells of Alcaligenes sp. AA09.
Acknowledgement
We would like to acknowledge the assistance provided by SRM University, Chennai for financial
assistance and Textile Engineering Department Anna University, Chennai for providing the dye sample.
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