Journal of Environmental Science, Computer Science ...

JECET; March 2015-May 2015; Sec.A Vol.4.No.2, 301-310

.E-ISSN: 2278–179X

Journal of Environmental Science, Computer Science and Engineering & Technology An International Peer Review E-3 Journal of Sciences and Technology

Available online atwww.jecet.org Section A: Environmental Science Review Article

Bioefficacy of Textile Effluent Adapted Bacterial Strains for COD Reduction Neha Sharma*, Sonika Saxena ,Aparna Datta, Saksham Gupta Dr B. Lal Institute of Biotechnology, 6-E, Malviya Industrial Area, Malviya Nagar, Jaipur(Rajasthan) India Received: 12 February 2015; Revised: 26 February 2015; Accepted: 04 March 2015

Abstract: Textile industry is one of the most important sectors which contributes to

Rajasthan’s economy and at the same generates huge volumes of waste water characterized by intense colour, high pH, BOD, COD. Keeping in view the above cited fact, an attempt has been made to characterize untreated textile waste water collected in accordance with standard procedures from a local dyeing unit based at Sanganer, Jaipur. objective of our study was to monitor COD reduction by microcosm analysis at an interval of 5 days till 20 days by indigenous bacteria and the strains was biochemically characterized as Enterobacter sp., Pseudomonas sp., Sarcina sp., Bacillus sp. and consortium was established. The strains were inoculated in Nutrient Broth (NB) to study growth curve. COD reduction was carried out by in situ analysis in which the inoculum concentration was varied from 0.1 % to 1 % v/v. 1% inoculum concentration was found to be most promising in terms of COD reduction. Keywords:Textile industry, Industrial effluents, ecological,pollutants,

JECET; March 2015-May 2015; Sec.A Vol.4.No.2, 301-310.

301

Bioefficacy ...

Neha Sharma et al.

INTRODUCTION Cleaner production is the need of an hour for sustainable development. Textile industry in particular becomes one of the culprits to be considered in terms of damage it causes to the environment. The strategic approach for cleaner production has gained momentum in recent years for environmental protection and preservation of ecological resources from excessive depletion and, has shown its ability to decrease environmental pollution, preserve natural resources Li et al 1.The current problem faced with industrial waste waters is absence of treatment prior to its being released into environment .Industrial effluents urban runoff, direct disposal of untreated textile effluent into adjoining water bodies specially the drains, fertilizers and animal wastes constitute the major reservoir of contaminants Jaishree and Khan 2. Excessive usage of enormous volumes of water and chemically different types of synthetic dyes contributes to water pollution caused by textile industry. The major constituents of textile based effluents are several types of chemicals such as dyes, dispersants, leveling agents, acids and alkali Olukanni et al 3 are discharged into water bodies without treatment which increases the chemical oxygen demand (COD) and biochemical oxygen demand (BOD), alters the pH and gives the water bodies (rivers) intense colorations. Coloured wastewater from textile industries is considered to be the most polluting in almost all industrial sectors Andleeb et al 4.Sanganer textile industry which is one of the largest handloom manufacturing zones from western India covers more than 154 block printing units. It isbecause of the toxic nature of effluents generated by textile cluster, it has gained much attention Talware et al 5. Effluent from these industries is being discharged without treatment into agricultural fields and used for irrigational purposes and pollutes both ground water and soil due to the percolation of some water soluble pollutants Pandey et al 6.COD being one the major pollution indicator reflects the quality of water both in terms of its use for irrigational purposes and release into adjoining surface waters. The fact has been established that untreated textile waste water has very high COD7-9 Sanganer being one of the major textile clusters of Rajasthan is deprived of an Effluent Treatment Plant (ETP),the waste water generated from textile and dyeing units is directly introduced into the adjoining surface water and primarily is used for irrigational purposes. Studies pertaining to reduction in COD by indigenous micro flora have been well established both nationally and internationally. Sharma et al.13 studied the effect of indigenous bacteria on simulated textile effluent in terms of COD reduction and dye degradation. Roy et al.14studied the effect of algae and aquatic macrophyteson COD reduction of textile effluent collected from post discharge of equalization tanks and witnessed decrease of 69% in COD. Musa andAhmad15 studied the effect of indigenous bacteria on waste water of pineapple industry in terms of reduction in COD by microcosm analysis. Keeping in view, the above cited facts, our study is aimed to characterize the untreated textile effluent and to isolate and identify a potential bacterial strain the effect of which would be studied to minimize the level of COD through microcosm analysis. MATERIALS AND METHODS

Sampling Site: Textile waste water samples were collected in accordance with standard procedures16 from a local dye house society situated as Sanganer, Jaipur.Figure 1 represents the sampling site and Figure 2 represents the textile waste water samples.

JECET; March 2015-May 2015; Sec.A Vol.4.No.2, 301- 310.

302

Bioefficacy ...

Figure 1: Sampling Site

Neha Sharma et al.

Figure 2: Textile waste water samples

Microbiological analysis: The textile waste water sample was serially diluted upto 10

-10

dilutions.

These dilutions were then quadrant streaked onto general purpose media (Nutrient Agar ) with the following composition (mg/l): Beef extract 3gm, NaCl- 5gm , Peptone -5gm; Agar: 15 gm and incubated at 37ºC for 24-48 hours. Isolation and identification of bacterial isolates: From the mixed culture obtained following incubation, individual colonies were picked up and quandrant streaked onto nutrient agar plates and were also grown in nutrient broth until a desired OD660 nm was attained

17

. The strains were

designated as Tex- A, Tex- B, Tex-C and Tex-D. Inoculum preparation for in situ studies: Different inoculum concentrations of all strains (Tex- A, Tex- B, Tex-C and Tex-D) were prepared as 0.1%, 0.5% and 1%v/v. In situ bioremediation of untreated textile effluent was carried out in accordance with protocol devised by Musa and Ahmad, 2010. The effluent sample was equally distributed (400 ml each) into a set of 3 erlenmeyer flasks (1000ml capacity) and labeled as I, II and III A separate flask containing only the effluent sample was maintained as negative or abiotic control .In flask I, 0.1 % (v/v) inoculums was added, to flask II, 0.5 % (v/v) inoculum was added and to flask III, 1 % (v/v) inoculums was added whereas flask IV was devoid of inoculum. A consortium was also prepared in which all the strains at 1 % (v/v) were inoculated in effluent sample.


303

Bioefficacy ...

Neha Sharma et al.

Bioefficacy of strains in terms of COD reduction: The simulated effluent samples were analyzed for COD reduction by dichromate reflux method at an interval of 5 days till a period of 20 days for all inoculum concentrations (0.1 %, 0.5 % and 1%) The decrease in COD was expressed in terms of percent reduction by the formula: Initial Final

% COD reduction =

COD X

100

Initial COD

RESULTS AND DISCUSSION

The bacterial isolates were designated Tex-A, Tex-B, Tex-C and Tex-D and were identified by their biochemical characteristics. The results of different biochemical properties of bacterial isolates are given in Table 1. Figure3 represents pure cultures of bacterial isolates. Biochemical characteristics of bacterial isolates: Table 1: Biochemical tests of bacterial isolates S.No. 1. 2

Tests Gram Stain Cultural Characteristics

3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

Lactose Dextrose Sucrose H2S. Production NO3 Reduction Indole Production MR Reaction VP Reaction Citrate Use Urease activity Catalase activity Oxidase Gelatin Starch Lipid Genus

(Tex-A) Rod (-) abundant, thick white, glistening growth AG AG AG ± + + + + Enterobacter Sp.

(Tex-B) Rod (+) Abundant, opaque, white waxy growth A A + ± + + (Rapid) + ± Bacillus Sp.


(Tex-C) Coccus(+) Pale orange colonies

(Tex-D) Rod (-) abundant, thin white growth, with medium

+ + + + + + + + + Sarcina Sp.

+ + + + + Rapid + Pseudomonas Sp.

304

Bioefficacy ...

Neha Sharma et al.

Figure 3: Pure culture of isolates on Nutrient Agar (Tex-A, Tex-B, Tex –C and Tex-D)

COD reduction: Reduction in COD Microcosm analysis was carried out by standardizing the O.D.660 =0.6 of the isolates. Three different inoculum concentrations were used (0.1%v/v, 0.5% v/v and 1%v/v). For all the strains under study, a maximum reduction in COD was observed with 1% v/v and least with 0.1%v/v for all strains. Figure 3 represents plot of reduction in COD by Enterobacter sp. with increase in incubation hour. A maximum reduction of 88.5% reduction in COD was observed after 20 days of incubation with 1%v/v inoculum. A minimum of 16.41% reduction in COD was attained on day 1 with 1% v/v inoculum. Reduction in COD attained by abiotic control was found to be in range 6.71% -10%. Similar trends in COD reduction by Sarcina sp.was observed. 1% v/v inoculum size reduced the COD of effluent sample by 80% on 20th day of incubation, and a minimum of 1.64% reduction in COD was observed with 0.1% v/v on day 1. Decolourisation by abiotic control was found to be in the range 1.04%-11.64% (Figure 4). Figure 5 represents COD reduction by Bacillus sp. A maximum of 73.43% of reduction in COD was observed on 20th day and a minimum of 12.32% with 0.1% v/v on day 1 of incubation. With abiotic control COD was reduced in the range 4.61%-11.94%. Figure 6 represents COD reduction by Pseudomonas sp.was 89.2% on 20th day of incubation and it was found to be maximum for all strains under study. A minimum of 16.41% of COD was found to reduce on day 1 with 0.1% v/v for the strain. For abiotic control, 0.59%-10.59% COD was reduced.


305

Bioefficacy ...

Neha Sharma et al.

0.1 % v/v 0.5%v/v

80 60 40

1.0%v/v

20

Abiotic control

0

1

100 COD % reduction

COD % reduction

100

0.5%v/v

40

1.0%v/v

20

40

0.5%v/v

30 20

1.0%v/v

10 Abiotic 5 10 15 20 control Incubation time(in days)

COD % reduction

Fig. 5: Bio efficacy of Bacillus sp with different inoculum % for COD reduction 90 80 70 60 50 40 30 20 10 0

COD % reduction

0.1%v/v

50

1

5 10 15 20 Incubation time(in days)

different inoculum % for COD reduction

70 60

Abiotic control

Fig.4: Bio efficacy of Sarcina sp.with

80

COD % reduction

60

0

different inoculum % for COD reduction

1

0.1% v/v


Fig. 3: Bio efficacy of Enterobactersp with

0

80

100 90 80 70 60 50 40 30 20 10 0

0.1%v/v 0.5%v/v 1.0%v/v Abiotic control 1


Fig. 6: Bio efficacy of Pseudomonas sp. with different inoculum % for COD reduction

I II III IV

1


Fig. 7: Bio efficacy of Cons T with different inoculum % for COD reduction


306

Bioefficacy ...

Neha Sharma et al.

Figure 8 represents comparative analysis of indigenous bacterial strains (1 % v/v inoculum) for COD reduction expressed as percent reduction by all strains. Maximum COD reduction of 89.2% was attainable by consortia (Cons T) after 20 days of incubation whereas a minimum of 18.2% reduction in COD was accomplished by Bacillus sp. on day 1.

100 90 COD Reduction %

80 70 Enterobacter

60

Bacillus

50

Sarcina

40

Pseudomonas

30

Cons T

20 10 0 0

5

10 Days

15

20

Fig. 8: Comparative analysis of indigenous bacterial strains (1 % v/v inoculum)for COD reduction expressed as percent reduction DISCUSSION Wastewater from industrial plants such as textile, electroplating and petroleum refineries can contain various substances that tend to increase the chemical oxygen demand (COD) of the wastewater. Various local agencies have placed limits on the allowable levels of COD in industrial wastewater effluent. It is desired to develop a process suitable for treating the wastewater to meet the regulatory limits. Various methods are available for COD reduction in industrial wastewater such as precipitation, incineration, chemical oxidation and biological oxidation. This study attempts to solve the high concentration of COD in industrial wastewater using bacteria. The organic matter that contributed to COD in the wastewater will be degraded by the bacteria under aerobic conditions using batch process. The wastewater will be supplemented with an agricultural liquid waste for bacterial growth and metabolism. This will ensure the continual presence of bacteria to carry out degradation of organic matter in the wastewater. The effect of bacterial consortium on COD reduction in dye contaminated soil sampled from vicinity of textile industry was studied by Sharma et al,13. A similar study in which indigenous bacterial strains have been used for COD reduction has been investigated in effluent generated from pineapple industry has also been reported JECET; March 2015-May 2015; Sec.A Vol.4.No.2, 301- 310.

307

Bioefficacy ...

Neha Sharma et al.

Musa and Ahmad15.It is expressed as percent reduction which is calculated at regular intervals of 5 days. Microcosm analysis with an aim to reduce COD of textile effluent by bacteria and fungi isolated from textile dye contaminated soil has been reported Agarry and Ajani10 Reduction in COD of textile effluent by establishing aerobic –anaerobic bioreactor has been well accounted18. Roy et al, 14 collected effluents samples from post discharge of equalization tanks of local composite textile mills of Saver were treated with aquatic macrophytes, algae and their combination. They were found to be effective for the reduction of chemical oxygen demand (COD) and pH. Sixty nine per cent of COD was reduced with the combination treatment of Nostoc, Eichhorniacrassipes and Pistiastratiotes. With the combination treatment of Nostocand E. crassipes reduced 65 per cent COD in glass containers. pH was reduced from 11.2 to 8.6. Between earthen and glass containers, glass container was found to be more effective. Wastewater that has been discharged from from the pineapple industry contributes to high levels of Biochemical Oxygen Demand (BOD), Chemical Oxygen Demand (COD) and suspended solids. The wastewater treatment using bacterial pellet showed higher COD reduction as compared to whole bacterial culture 15.Sharma et al,13 reported the potential of mixed bacterial culture for biodegradation of simulated textile effluent.COD was removed in aerobic process. Ajao et al.19 reported immobilisation of Pseudomonas aeruginosa and Bacillus subtilis found to have degradative capacity was immobilized on agar-agar, which was transferred into bioreactor fitted with air sparger. The effluent was treated in the bioreactor. The immobilized cells significantly reduced COD to 200mg/I, BOD to 20mg/l, TSS 300mg/I that are upper limit for disposal into surface water. The result indicates overall % reduction in COD, BOD, Nitrate, Sulphate, Phosphates as 83%, 97%, 61.3%, 62.8%, 61.2% respectively CONCLUSION

Primary objective of our research was based on bio augmentative approach aimed to reduce COD of untreated textile effluent. The autochthonous bacterial strains exhibited a tremendous potential to reduce COD by microcosm analysis consortium was also developed which was found to be most promising for COD reduction by microcosm analysis and through bioaugmentative approach the effect of the same on other pollution indicators like BOD, suspended solids could be studied and the process could be scaled up and in situ bioremedial approach. Also, the study could be extended to study the enzymatic mechanisms of the selected strains which were responsible for reducing COD up to appreciable level. Since, study of biodiversity offers an economic and cost effective tool for bioremediation of textile waste water, culture independent approach to assess the diversity of indigenous microflora by metagenomic analysis can be a promising alternative to combat water pollution caused by textile industries. ACKNOWLEDGEMENT

The authors would like to thank Dr. B. Lal Institute of Biotechnology for providing an excellent platform for carrying out the research. JECET; March 2015-May 2015; Sec.A Vol.4.No.2, 301- 310.

308

Bioefficacy ...

Neha Sharma et al.

REFERENCES 1. Z. Li, S. Zhang, Y. Zhang, L. Wei, Evaluation of cleaner production audit in pharmaceutical production industry: case study of the pharmaceutical plant in Dalian, P. R. China, Clean Techno Environ Policy 2011,13 (1): 195–206. 2. Jaishree, T.I.Khan, Monitoring of Heavy Metal in Textile Waste Water of Sanganer, Jaipur (Rajasthan). International Journal of Scientific and Research Publications 2014, 4(3): 1-4. 3. O. D. Olukanni, A. A. Osuntoki, G. O. Gbenle, Textile effluent biodegradation potentials of textile effluent-adapted and non-adapted bacteria, Afri. J. Biotechnol 2006, 5 (20) :1980 – 1984. 4. S. Andleeb, N. Atiq, M. I. Ali, R.L.Hussain, M. Shafique, , B. Ahmed, , P. B. Ghumro, , M. Hussain, A Hameed, S. Ahmed, Biological treatment of textile effluent in stirred tank bioreactor. Int. J. Agric. Biol 2010, 12 (2): 256 – 260. 5. P.P. Talware, V.S. Shrivastava, Metallic and organic study: A Detective green chemistry Approach, Asian Journal of Chemical and Environmental Research 2008, 1(4): 54-57. 6. K.S. Pande, S.D. Sharma, Distrubution of organic matter and Toxic Metals in the sediments of Ramganga River Moradabad, Pollution Research 2009,18, 43-48. 7. N. Sharma, S. Chatterjee, , P. Bhatnagar, Assessment of Physicochemical Properties of Textile Wastewaters and Screening ofBacterial Strains for Dye Decolourisation,Universal Journal of Environmental Research and Technology 2013, 3 (3): 345-355. 8. V. Goyal, Sudesh , S. Singh, Physico - chemical analysis of textile effluents of dye and printing clusters of Bagru region, Jaipur, India. Journal of Environmental Research and Development 2013, 8 (1): 11-15. 9. S. A. Paul, S. K. Chavan , S. D. Khambe, Studies on characterization of textile industrial

waste water in Solapur city. Int.J. Chem. Sci; 2012 10 (2): 635-642. 10. S. E. Agarry, A. O. Ajani, Evaluation of Microbial Systems for Biotreatment of Textile Waste Effluents in Nigeria: Biodecolourization and Biodegradation of textile Dye, J. Appl. Sci. Environ. Manage 2011, 15 (1): 79 – 86. 11. L. Varma, J. Sharma, Analysis of Physical and Chemical Parameters of Textile Waste Water, Journal of International Academy of Physical Sciences 2011, 15 (2):269-276. 12. K. V. B. Rao, A. Prasad, Physico chemical analysis of textile effluent and decolorization

of textile azo dye by Bacillus endophyticus strain VITABR13 The IIOAB Journal, Research: Bioremediation 2011, 2(2): 55-62. 13. P. Sharma, L. Singh, J. Mehta, COD reduction and colour removal of simulated textile mill wastewater by mixed bacterial consortium, Rasayan J. Chem 2010, 3(4): 731-735. 14. R. Roy, A. N. M. Fakhruddin , R. Khatun, M. S. Islam , Reduction of COD and pH of textile industrial effluents by aquatic macrophytes and algae, Journal of Bangladesh Academy of Sciences 2010, 34(1): 9-14. 15. Musa, N. S., and Ahmad, W.A. 2010. Chemical Oxygen Demand Reduction in Industrial Wastewater using Locally Isolated Bacteria Proceedings of Regional Annual Fundamental Science Seminar (RAFSS) 2010..http://www.ibnusima.utm.my/rafss ;2010.


309

Bioefficacy ...

Neha Sharma et al.

16. APHA (1998). Standard methods for examination of waters and wastewater 20th Edition American Public health Association Washington (APHA) American water works association (AWWA),Water environment Federation(WEF)Washington DC USA. 17. R. Suizhou, , G. Jun , Z. Guoqu, S. Guoping, Decolourization of triphenymethane, azo

and anthraquinone dyes by a newly isolated Aeromonashydrophila strain, Appl. Microbiol Biotechnol 2006, 72: 1316-1321. 18. Kassa R.M. 2007. Biological organic matter and nutrient removal from textile wastewater using anaerobic-aerobic bioprocess. School of Graduate Studies Environmental Science Programme.Thesis. Addis Ababa University, South Africa. 19. T. Ajao, G.B. Adebayo S.E. Yakubu, Bioremediation of Textile Industrial Effluent using mixed culture of Pseudomonas aeruginosa and Bacillus subtilis immobilized on agar agar in a Bioreactor. J. Microbiol. Biotech. Res 2011, 1 (3):50-56.

Corresponding Author: Neha Sharma; Dr B. Lal Institute of Biotechnology, 6-E, Malviya Industrial Area, Malviya Nagar, Jaipur-(Rajasthan) India


310

Journal of Environmental Science, Computer Science ...

Journal of Environmental Science, Computer Science ...

Suggest Documents