Decolorization of Acid Blue 25 Dye by Individual and Mixed Bacterial Consortium Isolated from Textile Effluents
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Int.J.Curr.Microbiol.App.Sci (2015) 4(5): 1015-1024 ISSN: 2319-7706 Volume 4 Number 5 (2015) pp. 1015-1024 http://www.ijcmas.com Original Research Article Decolorization of Acid Blue 25 dye by individual and mixed bacterial consortium isolated from textile effluents B.Aruna, L.Rathna Silviya, E.Shiva Kumar, P.Roja Rani, Durbaka V.R Prasad and D.VijayaLakshmi* Department of Microbiology, Yogi Vemana University, Kadapa-516003, India *Corresponding author A B S T R A C T The dye decolorizing bacterial isolates Bacillus sp1, Bacillus sp2, Acinetobacter sp., Citrobacter sp. and klebsiella sp. were isolated from the textile effluents. Different parameters such as carbon source, nitrogen source, temperature, pH and K e y w o r d s dye concentrations were optimized for decolorization of acid blue 25 dye by using selected bacterial isolates. The aim of the study was the decolorization of acid blue Anthraquinone 25 dye showed maximum 70% decolorization under optimum conditions. Among dye, Bio- all isolates the klebsiella sp. was found to be most efficient in dye decolorization. decolorization, All the selected isolates showed maximum decolorization in static condition Bacterial compared to shaking condition. The optimum pH obtained for decolorization of isolates, Acid acid blue 25 by bacterial isolates was 7-8 but Citrobacter sp. showed maximum blue dye 25, decolorization at optimum pH of 6-7 at 370C and in Citrobacter sp. it was at 250C. static condition Bacterial consortium i.e. combination of five bacterial isolates showed highest decolourization potential activity when compared to the individual isolates at different parameters. Maximum decolorization potential was observed in the presence of glucose as carbon source and yeast extract as a nitrogen source. Introduction Synthetic dyes have variety of colors and a discharging. Insufficient treatment of high stability to light, temperature and dyestuff wastes of the industries leads to microbial attack. Furthermore, some contamination of soil and natural water synthetic dyes such as Anthraquinone dyes bodies. Physical-chemical treatment are carcinogenic or mutagenic (Spadaro et processes have disadvantages where the al., 1992). (Poonam Dayaram et al., 2008,) contaminant is not destroyed, while reported that the synthetic and reactive dyes biological treatment methods are cheap and showed decolorization of textile effluents of offers the best alternative with proper dye contaminated soil. analysis and control the environmental pollution (Banat et al., 1996). One Synthetic dyes are widely used in the textile promising strategy is the use of microbes industries and their effluents are that possess the ability to decolourize 1015 Int.J.Curr.Microbiol.App.Sci (2015) 4(5): 1015-1024 synthetic dyes. Microbial decolourization isolates (Bacillus sp.1, Acinetobacter sp., and degradation is an environment friendly, Citrobacter sp.). But combination of five cost-competitive and alternative to physico- bacterial isolates showed highest chemical decomposition processes for the decolourization capability when compared treatment of industrial effluents (Verma and to other combinations like four and three Madamwar, 2003). Many bacterial and bacterial isolates. fungal species have the ability to absorb or degrade the textile dyes. Bacterial Materials and Methods decolourization of Anthraquinone dyes is either aerobic or anaerobic (Pandey et al., Sample collection 2007). Nature offers wide microbial community for decolorizing the textile dyes, Waste water along with soil samples anthraquinone dye decolourizing bacteria contaminated with textile effluents and can be isolated from soil and effluent water, dyeing industry were collected from the however their potential ecological niches discharges of Siera Silk Mills from were targeted. Bangalore. The water samples were collected sterilized screw cap bottles and Anthraquinone dyes represent the second soil samples were collected by polythene largest class of textile dyes extensively used bags. in the industries due to their wide array of color shades. They are resistant to Dye degradation due to their fused aromatic structure which remains for long time Acid blue 25 an Anthraquinone based dye causing considerable concern to the supplied by sigma Aldrich chemicals environmental pollution and also dye limited, water soluble and widely used in retention of the soils or effluents. textile industries. Its maximum absorption Bioremediation techniques have gained was 600-630 nm with the following much attention, microbial decolorization and chemical structure. degradation is cost- effective and eco- friendly compared to different conventional Medium methods. Some potential bacteria have the ability to decolorize synthetic / commercial The Bacterial cultures grown at 370C in the dyes used for textile dyeing. Effluents from basal medium viz Zhou and Zimmermann textile and dyeing industries cause serious medium (ZZ) containing the following in pollution to the environment. g/l, Glucose 0.5%,Yeast extract 0.5%, (NH4)2SO4- 0.5g/l, KH2PO4 - 2.66g/l, In the present study, we record to isolate Na2HPO4 - 4.32 g/l, dye - 100 mg/l. potential dye degrading bacteria and the development of bacterial consortium from Isolation and Screening of potential individual isolates. We tried with different strains combinations by mixing of five bacterial isolates (Bacillus sp.1, Bacillus sp.2, For isolation the textile effluents were Acinetobacter sp., Citrobacter sp. and collected as sources of microorganisms Klebsiella sp.) four bacterial isolates Cultures were obtained through serial (Bacillus sp.1, Bacillus sp.2, Acinetobacter dilution by pour plate method. Isolated sp., Citrobacter sp.) and three bacterial colonies were then obtained was preserved 1016 Int.J.Curr.Microbiol.App.Sci (2015) 4(5): 1015-1024 for the further use. Fresh culture was then culture supernatants at 600 nm using UV- inoculated in to nutrient broth and incubated Vis spectrophotometer. Percentage of for 24 hrs at 370C.In order to screen decolourization calculated by the following potential strain 2% 24 hrs fresh inoculums formula: was transferred in to the 100 ml of ZZ medium containing dye and incubated at D = [D0-D1/D0] x 100 370C.Potential strains utilizing dye as a nutrient source which were placed on ZZ Where, D=Decolorization in percentage, agar plates and incubated. From these D0=Initial absorbance, D1=Final absorbance. enriched cultures isolated colonies were repeatedly & subsequently inoculated on Results and Discussion nutrient agar to obtain potential cultures. Therefore strains that showed high Total Fourteen bacterial cultures were decolourization potential were chosen to be isolated, identified and screened for the for decolorization and degradation studies at decolorization degradation of different concentration, pH temperature & Anthraquinone dyes from textile effluent. other nutrient factors etc.. All the strains tested were further selected & screened for dye degradation on Screening and identification of dye morphologically and biochemically degrading bacterial isolates characterized for identification. Based on biochemical test they were identified as The organisms isolated on nutrient agar was Bacillus sp.1, Bacillus sp.2, Acinetobacter further sub cultured on modified ZZ medium sp., Citrobacter sp. and Klebsiella sp. were containing 100mg of acid blue 25 dye and selected for Acid blue 25 dye degradation. incubated at 37oC under static conditions. Dye degradation experiments were tested on After six days of incubation the selective medium i.e. ZZ medium and decolourization potential was calculated. decolorization potential was observed by The bacterial isolates utilizing the dyes as a five bacterial isolates individually and by the source of nutrient, showing high efficiency development of bacterial consortium of decolourization were selected for the (Nachiyar et al .2012 & Maulin p.Shah study. 2014). Bacterial consortium with combinations of five bacterial isolates Decolorization was studied on various (Bacillus sp.1, Bacillus sp.2, Acinetobacter carbon sources such as (Glucose, Fructose, sp., Citrobacter sp. and Klebsiella sp.), four Maltose, Sucrose & Mannitol), Nitrogen bacterial isolates (Bacillus sp.1, Bacillus sources (Yeast extract, Beef extract & sp.2, Acinetobacter sp., and Citrobacter sp.) Peptone), different dye concentration (100- and three bacterial isolates (Bacillus sp.1, 600mg/l) pH values (5, 6, 7,8and 9) and Acinetobacter sp., and Citrobacter sp.) were temperature (250,370,450C).Growth was tested and the percentage was recorded monitored by using spectrophotometrically. (Table 1). Among three combinations the most potential decolourization activity was After decolorization, the decolourized observed. With combination of five bacterial medium was centrifuged at 10000 rpm for isolates (Bacterial consortium) showed high 20 minutes to separate the cell lysate and decolourization percentage and further study supernatant. Decolourization potential was was carried. determined by measuring the absorbance of 1017 Int.J.Curr.Microbiol.App.Sci (2015) 4(5): 1015-1024 Effect of carbon source on dye decolorization in the presence of yeast decolorization extract. In combination of five bacterial isolates showed the decolorization activity Optimization of dye decolorization with five of Acid blue 25 with yeast extract was about bacterial isolates was analysed by the 86%, beef extract 56% and peptone 59% incorporation of Glucose, Sucrose, Fructose, under static conditions. Among all