Microbial technology for management of phenol bearing dyestuff wastewater

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Wal. Sci. Tecll. Vol. 33, No.8, pp. 47-5\. 1996. Copynght © 19961AWQ. Published by Elsevier Science Ltd Pnnted m Great Bntam. All nghts reserved. 0273-1223196 $15·00 + 0·00

Pergamon

PH: S0273-1223(96)00260-0

MICROBIAL TECHNOLOGY FOR MANAGEMENT OF PHENOL BEARING DYESTUFF W ASTEWATER Pradnya Kanekar, Seema Sarnaik and Anita Kelkar Division of Microbial Sciences, Agharkar Research Institute, G. G. Agharkar Road, Pune 41 1004. India

ABSTRACT Environmental pollution caused by dyestuff industry wastewater is two-fold since it contains toxic chemicals like phenol, aniline etc. and also imparts the colour of the dye manufactured. For management of dyestuff wastewater. microbial technology becomes a novel approach since microorganisms have the characteristic ability to break down aromatic compounds and decolourize the dyes. Microbial treatment of a dyestuff wastewater containing phenol and methyl violet was studied at the site of a dye factory near Pune in a fixed film bioreactor. The bioreactor consisted of a rectangular cement concrete tank of dImensions 9.5 m length x 7.3 m breadth x 1.5 m height and brick pieces of size 10 cm x 8 cm x 6.5 cm used as supporting media for formation of microbial film. A mixed culture of Pseudomonas alcaligenes (phenol degrader) and Pseudomonas mendocina (decolourizing methyl violet) grown in the dyestuff wastewater was spread on brick media. After formation of a microbial film on brick media. the dyestuff wastewater was recycled in form of a shower through the brick bed for 24. 48 and 72h. The influent loading rate was 1.44 m 3/m 2 brick bed area per day. The phenol and Chemical Oxygen Demand (COD) loading rates were on average 0.9 kg phenol/m 2/day and 5.67 kg COD/m 2/day respectively. The performance of the microbial degradation evaluated for a period of 68 days showed 97.5%. 63%. 54.4% and 51% removal in phenol. methyl violet (MV) content, COD and Total Organic Carbon (TOC) respectively in 24 hours of recycling and marginally enhanced in 48 and 72 hours. The microbial technology was thus effective in removal of phenol and the dye from dyestuff wastewater. Copyright © 1996 IAWQ. Published by Elsevier Science Ltd.

KEYWORDS Microbial treatment; fixed film bioreactor; Pseudomonas sp.; dyestuff wastewater; phenol; methyl violet.

INTRODUCTION In India. the dyestuff industry is well established and contributes significantly to the environmental pollution. Pollution of water caused by dyestuff industry wastewater is two fold since it contains toxic chemicals e.g. aniline and phenol which are used as raw materials and imparts colour of the dye(s) manufactured. It is therefore imperative to treat such wastewaters before discharging into the receiving waters. Microbial treatment of dyestuff wastewater becomes a novel approach since some microorganisms are endowed with the property to break down aromatic compounds into simpler intermediates and end products and decolourize the dyes. 47

P. KANEKAR et al.

48

Some reports are available in the literature on microbial degradation of phenol, aniline, dyes and their wastewaters. Evans et al. (1951), Gibson (1968), Bayley and Wigmore (1973), Murray and Williams (1974) and Anselmo and Novais (1984) have reported microbial degradation of aromatic compounds including phenol, catechol and cresols. Anson and Mackinnon (1984) have described aniline degradation. Biodegradation of dyes and dyestuff wastewaters has been reported by Brown (1981), Yatome et al. (1981), Plat et al. (1985), Bumpus and Brock (1988) and Kanekar and Sarnaik (1991). Since Pseudomonas spp., although versatile in degrading aromatic compounds, do not form settleable sludge, attempts have been made by some workers at immobilization of these bacteria on some supporting media which could be used in fixed film bioreactors. There are some reports on such fixed film reactor processes for degradation of phenolic compounds (Beltman and Rehm, 1985; Jeter, 1985; Westmier and Rehm, 1985 and 0' Reilly and Crawford, 1989). This paper describes microbial treatment of phenol bearing dyestuff wastewater in a fixed film reactor using cultures of Pseudomonas sp. at the site of the factory near Pune, manufacturing the basic dye methyl violet using dimethylaniline and phenol as the major raw materials. MATERIALS AND METHODS Chemical analysis of the waste effluent The dyestuff waste effluent was analyzed for pH, Chemical Oxygen Demand (COD), Phenol (Using Gas Chromatograph) and Total Organic Carbon (TOC, using TOC analyzer) as per standard methods (Greenberg et al., 1992). The dye, methyl violet (MV) was estimated by a spectrophotometric method by measuring optical density (O.D.) at 580 nm (the max value for MV) on Spectrophotometer DU 8B (Beckman, USA) as described by Kanekar and Samaik (1991). Gas chromatographic analysis of phenol was done using microcomputer based gas chromatograph and data station (CIC, Baroda, India) equipped with flame ionization detector (FID) and a stainless steel column (2 m x 3 mm) SE-30 (10% SE 30 on chrome W -AW, 80/100 mesh). Argon was used as carrier gas at the flow rate of 30 mUmin. and a mixture of hydrogen (30 mUmin) and air (300 mUmin.) for flame. The temperatures of the column, injector and detector were 180, 220 and 2200C respectively. Phenol (Sigma Chemicals, U.S.A.) was used as the standard at a concentration of 500 mgIL. Developinc microbial culture for treatment of the dyestuff wastewater Microbial cultures were developed by enrichment technique using cattle dung as the source of microorganisms and by adaptation of the culture to the wastewater as described by Kanekar and Sarnaik (1991). The microorganisms capable of growing on the wastewater were identified as Pseudomonas alcaligenes and Pseudomonas mendocina with reference to Bergey's Manual of Systematic Bacteriology (Krieg and Holt, 1984). The two species were tested for their ability to degrade the wastewater with special reference to removal of phenol and methyl violet, the major pollutants of the wastewater by growing them in the wastewater and estimating the phenol and MV content after 48 h of incubation under aeration at room temperature (28 +/- 2OC). Preparin~

inoculum of the culture

The two cultures grown in the dyestuff waste effluent and having inoculum density of I x 10 7 cellslmL were mixed in I: I proportion and multiplied further in the wastewater in a rectangular mild steel tank of 1.95 m 3 capacity (2.45 m x 1.27 m x 0.62 m) under aeration. This was designated as mother culture tank and the culture was maintained in it.

Dyestuff industry wastewater Settin~

49

up an on-site fixed film bjoreactor

The fixed film bioreactor was developed in a rectangular cement concrete tank of dimensions 9.5 m length x 7.3 m breadth x 1.5 m height at the site of the factory. The tank was filled with brick pieces of size 10 cm x 8 cm x 6.5 cm upto the height of 0.75 m from the bottom of the tank. Above this basal layer of brick media. two horizontal rows of brick pieces parallel to the breadth of the tank were raised. The tank was filled with a total volume of 69.35 m3 of the wastewater so as to keep 9.5 m x 7.3 m x 0.25 m column of the wastewater above the surface of the basal layer of the brick media. A mixed culture of Pseudomonas alcaligenes and Pseudomonas mendocina grown in the wastewater in the mother culture tank was spread on the brick media using a liquid circulation pump for 15 days. Formation of film of the culture applied was checked by resolution of the cultures from brick pieces collected at various depths in the reactor and identification in the laboratory. After the formation of microbial film. the dyestuff wastewater was recycled using a liquid circulation pump and in form of a shower through horizontal PVC pipes having holes of diameter 0.5 mm. The PVC pipes were raised on top of the two brick rows parallel to the length of the brick rows. Operatjonal conditions The influent loading rate was 1.44 m 3/m 2 brick bed area per day. The phenol and COD loading rates were an average of 0.9 kg phenoVm 2/day and 5.67 COD/m2/day respectively. The microbial treatment process was a batch process and the experiments were conducted for a period of 68 days. The samples of treated effluents drawn after mean residence times of 24. 48 and 72 hours were analyzed for pH. COD. Phenol. TOC and MV content. RESULTS AND DISCUSSION The data collected on biodegradation of the wastewater with special reference to the ability of the two microbial species to remove phenol and colour of the wastewater have been presented in Table I. It is seen from the table that while P. alcaligenes was efficient in removing phenol. P. mendocina was better in decolourizing the dye methyl violet from the wastewater. Table I. Biodegradation of the wastewater by Pseudomonas sp. Parameter

pH

MV, mglL

removal Phenol, mgIL % removal COD, mg/L % removal TOe, mglL % removal

Uninoculated control 7.60 2.60

%

99.74 6608 282.38

P. alcaligenel 7.14 0.70 73.08 0.0 100.0 1888 71.43 45.14 84.01

P. mendocina 7.32 0.45 82.69 24.45 75.49 2340 6459 50.34 82.17

The data collected on performance of the fixed film bioreactor with respect to removal of COD. Phenol. TOC and MV content are presented in Table 2 and Figure 1. The table and figure illustrate efficiency of the microbial process in removing phenol and colour of the wastewater. It is also seen that the performance of the bioreactor was marginally enhanced except for removal in TOC with extended residence time from 24 hours to 48 and 72 hours. Hence recycling of the wastewater through microbial film for a period of 24 hours was recommended to the factory authorities. The microbial process has been in operation at the site of the factory for the last two years.

so

P. KANEKAR el at. Table 2. Performance of the fixed film bioreactor in the treatment of the wastewater Period Parameter

24

U pH

range average

MV,

range

m~

average

T

or

recycling, h 48 T U

7.15 to 8.27 7.7

4.26 to 11.5 9.25

7.99

8.77

7.86

2 to 10.1 4.3

0.1 to 3.4 1.6

2.4 to 11.4 5.9

1 to 3

2.2 to 8.5

0.8 to 4.9

1.9

5.9

63.27

removal

61.03

range

27.2 to 2236

o to 113.7

36.6 to 2173

average

618

15.48

110

m~

7.25 to 8.91

%

3.21 to 10.8

T

4.01 to 11 .8 9.11

% Phenol,

72

U

97.5

7.05 to 8.41

1.8 69.25

o to 320.0

54.9 to 2324

25.67

704

96.38

o to 25.64 4 99.4

removal range

2711 to 7111

average

3919

COD, m~

%

1086 to 2104 1787

1922 to 9612

934 to 3270

2450 to 7445

3751

1787

4308

54.4

52.4

980 to 2127 1709 60.32

removal TOe, mg/L

range average

269 to 359 314

%

138.5 to 167.6

96 to 861

11 to 247

321 to 1479

163.9 to 212.1

153

470

116

900

188

51

75

removal

The values are average of 15 samples analyzed U c Untreated T '"' Treated

100 90

80 70 60 E 50

~ 0

~

. ~

ItO

30 20 10 0

21t ~MV

1t8 rtcycling period , hr" ~ PHENOL ~ COD

D10C

Figure I . Performance of the fixed film bioreaclor

79

Dyestuff industry wastewater

51

CONCLUSION The microbial technology has been found to be suitable for management of phenol-bearing dyestuff wastewater.

ACKNOWLEDGEMENT The authors are thankful to Mr R. D. Shembekar. Manager. Ganesh Dykem factory for all the necessary help to conduct the experiment at the site of the factory.

REFERENCES Anselmo. A. M. and Novais. J. M. (1984). Isolation and selection of phenol degrading microorganisms from an industrial effluent. Biotechnol. utters 6(9). 601-606. Anson. J. G. and Mackinnon. G. (1984). Novel Pseudomonas plasmid involved in aniline degradation. Appl. Environ. Microbiol. 48. 868-869. Bayley. R. C. and Wigmore. G. J. (1973). Metabolism of phenol and cresols by mutants of Pseudomonas putida. J. Bacteriol. 113. 1112-1120. Baltmann. H. and Rehm. H. J. (1985). Continuous degradation of phenols by Pseudomonas putida P8 entrapped in polyacrylamide hydrazide. Appl. Microbiol. Biotechnol. 22. 389-393. Brown. 1. P. (1981). Reduction of polymeric azo and nitro dyes by intestinal bacteria. Appl. Environ. Microbiol.41(5). 1283-1286. Bumpus. J. A. and Brock. B. J. (1988). Biodegradation of crystal violet by the white rot fungus Phanerichaete chrysosporium. Appl. Environ. Microbiol. 54(5).1143-1150. Evans. W. C .• Smith. B. S. W.• Lmstead. R. P. and Elvidge. J. A. (1951). Chemistry of the oxidative metabolism of certain aromatic compounts by microorganisms. Nature 168. 772-775 Gibson. D. T. (1968). Microbial degradation of aromatic compounds. Science 161.1093-1097. Greenberg. A. E.• Clesceri. L. S. and Eaton. A. D. (ed) (1992). Standard Methods for the Examination of Water and Wastewater. 18th edition. published jointly by APHA. AWWA and WPCF. Washington. D.C. Jeter. J. H. (1985). Biological treatement of phenolic paintstripping wastewater; Proceedings of the 40th Industrial Waste Conference. Purdue University. Indiana. Section 7- Paint. Ink and Dye waste. page 159. Kanekar. P. and Samaik. S. (1991). An activated sludge process to reduce the pollution load of a Dye-industry waste. Environ. Pollut. 70. 27-33. Krieg. N. R. and Holt. J. G. (eds) (1984). Bergey's Manual of Systematic Bacteriology VoLl. Williams and Wilkins. Baltimore. USA. Murray. K. and Williams. P. A. (1974). Role of catechol and the methyl catechols as inducers of aromatic metabolism in Pseudomonas putida. J. Bacteriol. 117(3). 1153-1157. O'Reilly. K. T. and Crawford. R. L. (1989). Kinetics of p-cresol degradation by an immobilized Pseudomonas sp. Appl. Environ. Microbiol. 55. 866-870. Plat. W.• Hadar. Y. and Chet. J. (1985). The decolourization of the polymeric dye poly-blue (polyvinylamine sulfonate anthroquinone) by lignin degrading fungi. Appl. Microbiol. Biotechnol. 21.394-396. Westmier. F. and Rehm. H. J. (1985). Biodegradation of 4-chlorophenol by entrapped Alcaligenes sp. A7-2. Appl. Microbiol. Biotechnol. 22. 301-305. Yatome. C .• Ogawa. T .• Koga. D. and Idaka. E. (1981). Biodegradibility of azo and triphenylmethane dyes by Pseudomonas pseudomallei 13 NA. J. Soc. Dyers Col. 97.166-169.