http://annalsofrscb.ro 1448
GCMS analysis of tannery effluent by using Halophilic Bacterial Strain Pseudomonas aeruginosa sthc002 and
Keratinase Enzyme
Aji R1 and Raja Jeya Sekar R2
1 Reg. No: 11109, Department Of Zoology, S.T. Hindu college, Nagercoil, Affliated to Manonmanium Sundaranar University.
2 Asst. Professor in Zoology, S.T. Hindu College, Nagercoil.
Abstract
The discharge of untreated effluents during the starting of new industries and development of the industries results in water, air and soil pollution. During this period industrial waste effluent considered as a pollution causer because of the disposal of untreated effluent. Water pollution is the major pollution that effects the environment much and one of the sources of this pollution is tannery industrial effluents. Effluents are so toxic that fishes cannot survive in it even for two hours and also effects the drinking water and hence it should be treated effectively before release. Bioremediation is the only way to tackle these xenobiotics and reduce the pollutants which is eco-friendly. Keratinase enzymes and halophilic microorganisms are ubiquitous in nature and usually found in all living organisms. This paper based on the characterization and GCMS analysis of tannery effluent by using halophilic bacterial strain Pseudomonas aeruginosa sthc002 and keratinase enzyme. An experiment were conducted to isolate keratinase producing microbial strain from tannery effluent from Puthalam salt pan, Kanyakumari District, Tamil Nadu. Biochemical tests were done to identify the microorganisms.
Key words: Keratinase,P. aeruginosa sthc002, tannery waste, GCMS, Waste degradation Introduction
Tannery effluents are the major reason for the pollution in environment[1]. In leather industry Chromium sulphate is the pollution causing agent[2,3]. Indian progress and elevated life styles of individuals are the indicators of industrialization and economic development[4,5]. However, industrialization brought many environmental hazards like land, water, soil and soil solid pollution[6,7]. All the sectors like industry, agriculture, construction, transportation, mining and consumers of our society generate waste materials[8]. Industries have been regarded as a major source of pollution due to inappropriate treatment of waste disposal methods[9]. The environmental pollutions results in the discharge of untreated effluents from the industries which harmfully effect the assimilation capability of the agriculture[10]. The quantity and the toxic materials in the waste effluent changes with the process in the industries[11]. Water pollution is a major pollution that effects the environment much and one of the main sources is industrial effluents[12]. The inherent property of the industrial technique is such that heavy quantities of water is consumes[13]. Tannery waste is a major hazard that affects the drinking water and hence it should be treated effectively beforedisposing.Chrome tanning is the major pollution by leather industry[14]. Bacillus subtilis are a great interest for the biotechnology. Bacillus subtilis is a gram positive organism found in soil and GIT of ruminants and humans[15,16]. These were
http://annalsofrscb.ro 1449 popular worldwide before the introduction of antibiotics[17]. By using the Bacillus subtilis, we are converting the harmful tannery effluents to less harmful[18].
The enzyme keratinaseis one of the all rounder, because it involves in the different biological reactions like cellular and organism level and also in nitrogen circulation naturally.Exoprotease (cleaves at the end of the polypeptide chain) and endoprotease (cleaves inside the chain) are the two classes of protease enzyme, based on the site where cleavage happens (hydrolytic enzyme).
This enzyme may divided into 7 groups; ser, cys, thr, aspartic keratinases., glutamic keratinases, metallo proteases and asparagine peptide lyases depends on the sequence of amino acid at active sites of enzyme. On the basis of evolutionary relationshipsbetween the proteolytic enzymesera of sequences are introduced for upgrading system of classification. Establishment of MEROPS database, in that peptidases are arranged into clans of 62 and families of 268, according tothe classification based on structur[19]. Bacillus genus includes the strains that produce serine keratinases, industrially. For various reasons the genus has major part in white biotechnology–
well established, non-pathogenicspecies (for example Bacillus cereus, Bacillusstearothermophilus, Bacillus licheniformis, Bacillus subtilis ) and into the culture medium multiple extracellular proteins are released. Years ago, interest improving, with culturing methodology, manipulation and genetic engineering were the factors supporting the adaptation mechanism of halophilic microbes to environment. Highsalinity adaptation can be acquired byvarious methods for controlling the osmotic stress. Salt-in process include the accumulation of KCl=NaCl in environment utilized by haloarchea, and studied about that, its enzymesneed 4-5M salt. Salt-out technique is another one, which accumulate or produce de novo solutes excluding salts. It is mainly utilized by halophilic bacteria and eukarya. Some halophiles associated with adaptive mechanisms may also work[20].
The analytical method GC/MS analysis, it identifies different substances within a liquid sample. Gas chromatography mass spectrometry analysis which identifies the unknown samples in the tannery effluent. Representation of industrial catalysts share can be done by the enzymes secreted by microorganisms. Several applications of keratinases makes them as major enzyme types. Here in this study we analyzed both raw and treated tannery effluent and find out the known and unknown components in the sample[21,22].
Methodology
Collection of Brine Sample from the Salt Pan
The experimental brine was collected from the reservoir and condenser pond from Puthalam salt pan, Kanyakumari District, Tamil Nadu, India. The brine sample was collected aseptically and transported to the laboratory for the isolation of enzyme producing microorganisms.The pH and salinity were recorded.
Isolation of Bacterial Strains
Bacterial strains were isolated by serial dilution technique. For stock solution 1000 microliter of sample was diluted to 10ml with deaminized water. Six test tubes each with 9ml of distilled water were set, marked as 1-6. 1 milliliter from stock solution transfer into first test tube without contamination making the dilution 10-1,the tube were shaken well to mix the contents properly.
1ml from 10-1tube was transferred to next test tube marked it as 10-2. The process were continued
http://annalsofrscb.ro 1450 till to get 10-6 dilution. From each test tube 100 µl samples was poured to sterile petri plates having halophilic agar and spread properly. Then it kept for incubation at thirty seven °C for 24hours.The bacterial strains were maintained by sub culturing every 3 weeks on slants containing Nutrient Agar. After incubation the culture plate was examined to confirm the presence of microbes on the plate. The isolated colony on the plate were subcultured to get pure culture of the specific microorganism.
Identification of Bacteria
Various tests were conducted to identify the bacterial strains. Enzymatic activities of microorganisms are widely used to differentiate and characterize bacteria. Related bacteria can be separated into distinct species by using biochemical tests. The isolated organisms were subjected to the following biochemical tests and were identified by comparing with the standard data provided in Bergey’s manual of Systematic Bacteriology (2000).
Colony Morphology
Colony morphology was the initial step in identifying a bacterium. The colony morphology of the bacterial culture grown on the halophilic agar was examined for their size, colour, shape, margin and elevation.
Screening of Extracellular Enzyme Producing Bacteria
The extracellular enzymes protease and keratinase producing bacteria were screened with the help of suitable medium.
KeratinaseEnzyme Producing Bacteria
For assessing the keratinase activity, the bacterial strains were streaked on skim milk agar plates and incubated at 37 oC for 24 hours for production. The milk consist of casein, which converted to small particles by keratinase enzyme. The cells producing keratinase would give a clear transparent zone around them in an otherwise opaquemilk media.
Optimization of Bacteria for Extracellular Enzyme Activity
The bacterial strains were optimized with physico chemical and nutritional source for maximum production of extracellular enzyme.
Effect of pH
The effect of different pH ranging from 5 to 11 was examined for the growth efficiency of the bacterial isolates. Nutrient agar medium added with NaCl were used as suitable for culturing bacterial strains. The incubation temperature was maintained at 37°C. The pH was adjusted by using 0.01 M Hcl and 0.01M NaOH.
Effect of Temperature
The isolated bacteria were studied at different temperature such as 30°C, 35°C, 40°C, 45°C, 50°C and55°C. Each bacterial strain was optimized with different temperature to produce maximum extracellular enzyme.
http://annalsofrscb.ro 1451 Effect of Incubation Time
The incubation time varying from 24-120 hours were examined for the detection of optimum incubation time required for the growth of the bacteria. The incubation temperature was maintained at 37°C.
Effect of Salt Concentration
The effect of salt concentration on optimization of isolates was determined by increasing of NaCl concentration from 1 to 8 % concentration of NaCl was incubated at optimized conditions of all the above parameters understatic condition.
Effect of Carbon Source
The effect of several sources of carbon such as paddy glucose, fructose, sucrose, xylose and starch were used to study the growth of the bacterial strains and their respective extracellular enzyme activity. In 100ml of nutrient agar medium, 1% of respective carbon source was added individually. The incubation temperature was maintained at 37°C.
Effect of Nitrogen Source
The effect of various organic nitrogen sources such as beef extract, yeast extract, peptone, gelatin, casein and urea were used to study growth of the bacterial strain and their respective extracellular enzyme activity. In 100ml of nutrient agar medium 1% of respective nitrogen sources were added individually. The incubation temperature was maintained at 37°C.
Effect of Metal Ions
To evaluate the effect of various metal ions on the growth of bacterial strains and extracellular enzyme activity the metal ions such as sodium nitrate, ammonium sulphate, ammonium chloride, ferrous sulphate, calcium chloride and magnesium chloride were used. Each of the metal ions was separately added at 1 mM concentration in the 100 ml nutrient agar medium to study the growth of the bacterial strains and extracellular enzyme activity.
Biomass Production by the Bacteria
Pre-dried and pre-weight measure filter paper were placed on a petriplate .Samples were added filter paper placed in petriplates. Then dried at 105°C overnight in a hot air oven. After drying the final weight was determined by using an analytical weighing balance.
Tannery Effluent Collection
In the present study, tannery wastewater was collected from common effluent treatment plant, Pallavaram, Chennai. Samples were collected in sterilized glass bottles aseptically and transported to the laboratory in an ice pack condition. The collected samples were preserved at 4°C in refrigerator for further analysis.
Physico-chemical Characterization of Samples
http://annalsofrscb.ro 1452 In the present study, tannery wastewater was collected from common effluent treatment plant, Pallavaram, Chennai. The collected tannery effluent sample was used to study parameters like pH, temperature, Biological Oxygen Demand, Chemical Oxygen Demand,Total Dissolved Solids,chloride and hardness in order to determine the quality of raw textile effluent, bacterial strain treated effluent and extracellular enzyme effluent.
pH
pH meter is the equipment used to find the pH. The effluent sample was added to a conical flask (100 ml) and the pH was measured. The pH electrode was rinsed with distilled water and calibrated using standard solution before use.
Temperature
Temperature is a measure of heat in terms of a standardized unit. It is an important ecological factor and an environmental variable, not just seasonal- as it also fluctuates on daily or even hourly bases.
Biochemical Oxygen Demand (BOD)
DO determination of one group of flasks has been done, at the 5 days incubationtime (20°C) of second group of flask.Then the incubated sample of DO was determined after 5 days. To the contents of the BOD bottle 2 ml of manganoussulphate solution and 2ml solution of alkali-iodide azide was added. Thoroughly,the bottle was stopped and gently mixed. There is a formation of precipitate which is brown in color called basic manganic oxide, settle down the precipitate for the formation of clear supernatant.For completing dissolution process few ml of conc.H2SO4
were added to the sides of bottle, and then stopped, gently mixed. The titrationof this contents against sodium thiosulphate (0.025N) and utilize indicator starch. The titration was continued until it becomes colorless.
(DO0–DO5 –BC) X Volume of diluted sample BODmg/L = ---
Volume of the sample taken
For titration process, the volume of sodium thiosulphate (0.025N) used = DO in mg/l DO0 = Initial DO
DO5 = DO after 20 ºC incubation for 5 days
BC = Blank correction (Difference in DO of blank on the initial day and after 5 days incubation Chemical Oxygen Demand (COD)
A 250mlof refluxing flask was fitted with double surface condenser having 300mm into which glass cap was fitted. Into this flask 50ml of the sample was transferred. 1gm of mercuric sulphate, few glass beads and 5ml of sulphuric acid-silver sulphate reagent was added, mixed well and allowed to cool. 25ml of 0.125N potassium dichromate was pipetted into the flask. The flask was connected to the condenser and cooling water was turned on. 70ml of sulphuric acid
http://annalsofrscb.ro 1453 reagent was added through open end of condenser with swirling and mixing. Water circulation was started and refluxed for 2hrs. For dilution process, the contents were added to 350ml of distilled water. It titrated against solution of FAS (0.125N) after adding few drops of indicator ferroin. The color turned into reddish brown from blue green. A blank with distilled water instead of sample along with reagents was used. The same procedure was followed as for the sample.
(Titre value of blank –Titre value of sample) x 0.125 x 1000 x 8 COD in mg/L = ---
Volume of the sample taken Total Dissolved Solids (TDS)
250ml of the effluent sample was filtered using a glass microfiber filter paper. Then the filtrate were boiled in a tared porcelain dish , preheated at 105°C and then at 550°C for in a 1 hrs in muffle furnace and cooled. The dish was kept at 180°C for one hour, cooled and weighed.
(A-B) x 1000 Total dissolved solids (TDS) mg/L = ---
Volume of sample taken A = weight of dried residue + dish, mg
B = weight of dish, mg Estimation of Chloride
20 ml of sample was taken and the pH was adjusted to 7-9.5. Titration against solution of standard silver nitrate with continuous stirring after adding potassium chromate indicator of 1ml, till red color formed in precipitate. The titration was repeated with 100ml blank containing distilled water (chloride free) instead of sample.
(A-B) x N x 35.45 x 1000 Chloride mg/L = ---
Volume of sample taken A = Volume of silver nitrate for sample
B = Volume of silver nitrate for blank N = Normality of silver nitrate
Total Hardness
20 ml of tannery effluent placed in a beaker that requires 20 ml EDTA titrant (0.02 M) and after adding buffer, within 5 minutes titration have to be complete. Using distilled water 25ml have to be diluted to 50ml. For getting 10.0 to 10.1 pH, 1 to 2 ml buffer solution have to add. And also provide few drops of indicator solution of Erichrome Black T or approximate amount powder.
Until, the disappearance of red tinge, slowly add titrant of EDTA, with constant mixing. At 3 to 5 seconds of interval, last 2 to 3 drops have to be added. The solution is blue at end point.
A xB x N x 1000
Hardness mg/L = --- Volume of sample taken A: Volume of EDTA
N: Normality of EDTA
B: Equivalent weight of CaCO3
http://annalsofrscb.ro 1454 Chromium Removal
Chromium content of tannery wastewater was determined before and after the experiment. The candidate bacterial species was inoculated (1%, v/v) and incubated in tannery wastewater at room temperature (30 ± 2 °C) for seven days. The wastewater was first digested using HNO3
followed by colorimetric determination using a UV- visible spectrophotometer at 357.9 nm (Clesceriet al., 1999).
Influence of Bacterial Strain on Biodegradation of Tannery Effluent
The rate of degradation expressed in percentage of effluent treated with bacterial strains.
Absorbance reduction at absorption maxima was observed for the determination of percentage.
Effluent sample contained in un inoculated nutrient medium was taken as a reference. Reaction mixture samples of 2 ml were centrifuged at 10,000 rpm. For separating biomass 10 minutes is required. At 660 nm, the determination of dye concentration has to be done. Spectrophotometer is used for measuring absorbance. Depends on the ratio of percentage the efficiency of removing color was stated, following the equation.
Initial OD−Final OD ×100 Dye Degradation (%) = ---
Initial OD
Influence of Extracellular Enzymes on Biodegradation of Tannery Effluent
The rate of degradation expressed in percentage of effluent treated with extracellular enzymes.
Absorbance reduction at absorption maxima was observed for the determination of percentage.
Effluent sample contained in un inoculated nutrient medium was taken as a reference. Reaction mixture samples of 2 ml were centrifuged at 10,000 rpm. For separating biomass 10 minutes is required. At 660 nm, the determination of dye concentration has to be done. Spectrophotometer is used for measuring absorbance. Depends on the ratio of percentage the efficiency of removing color was stated, following the equation [16].
Initial OD−Final OD ×100 Dye Degradation(%)= ---
InitialOD
GC-MS
GC-MS of metabolites was carried out by using PerkinElmer (UK) equipped with a PE auto system XL gas chromatograph and a PE-5MS capillary column (20 m × 0.18 mm internal diameter, 0.18 mm film thickness). Helium was used as a carrier gas (flow rate: 1 mL min-1) using split less injector (injector temperature was 280 °C). The column temperature was programmed as 50 °C (5 min); 50-300°C (10 °C min-1, hold time: 5 min). The MS transfer line and ion source temperatures were kept at 200 and 250 °C, respectively. A solvent delay of 3.0 min was selected. In full-scan mode, the electron ionization (EI) mass spectra were recorded in range of 30-550 (m/z units) at 70 eV. Retention time and fragmentation patterns were analysed.
Data Analysis
http://annalsofrscb.ro 1455 The data were analyzed the present study, following the statistical analysis described by Zar (1974).
Standard Déviation (SD) SD = √ (Σd2)
N Where,
d- Refers to the deviation of each scope from mean N- The total number of samples.
Analysis of Variance (ANOVA)
One way and two way ANOVA were found with the help of the software PASW statistics data editor and Ky plot respectively. Means were compared at 0.05 % for Two Way ANOVA and 0.001 % level.
Results and Discussion Isolation of Bacterial Strains
Keratinase Enzyme Producing Bacteria
The halophilic bacterial strain P. aeruginosa sthc002 streaked at the centre of the sterile skimmed milk agar plate showed a dumb bell shaped zone around the bacterial colonies at 37 °C and 24 hours of incubation, it was shown in fig: 1.
Fig 1: Screening of P. aeruginosa sthc002 (Accession number: MW332065) for Keratinase activity
Optimization of Bacteria for Extracellular Enzyme Activity
http://annalsofrscb.ro 1456 Bacterial strains were optimized with nutritional source for maximum production of extracellular enzyme.
Influence of ph and Temperature Ph 8, temperature 40 °C
Influence of NaCl Concentration on P. aeruginosa sthc002 AJ for Keratinase Production Max produ 4%, Enzyme production 68.3 ± 0.05 U / ml. Biomass 22.5 ± 0.05 mg / ml
Min produ 2%, Enzyme production 14.0 ± 0.03 U / ml. Biomass 4.5 ± 0.03 mg / ml Influence of Incubation Time on P. aeruginosa sthc002 AJ for Keratinase Production Max produ 72hr, Enzyme production 69.4 ± 0.02 U / ml. Biomass 24.4 ± 0.04 mg / ml Min produ 2%, Enzyme production 19.0 ± 0.04 U / ml. Biomass 5.2 ± 0.03 mg / ml
Influence of Carbon Sources on P. aeruginosa sthc002AJ for Keratinase and Biomass Production
Glucose, maltose, fructose, xylose and starch
Max produ maltose, Enzyme production 65.2 ± 0.03 U / ml. Biomass 22.2 ± 0.06 mg / ml Min produ glucose Enzyme production 51.5 ± 0.01 U / ml. Biomass 15.5 ± 0.02 mg / ml
Influence of Nitrogen Sources on P. aeruginosa sthc002 AJ for Keratinase and Biomass Production
Beef extract, yeast extract, casein, peptone, gelatin
Max produ –Yeast extract, Enzyme production 83.3 ± 0.03 U / ml. Biomass 17.8 ± 0.03 mg / ml Min produ- beef extract Enzyme production 57.3 ± 0.03 U / ml. Biomass 8.3 ± 0.06 mg / ml Effect of Metal Ions on P. aeruginosa sthc002 AJ for Keratinase and Biomass Production Sodium nitrate, ammonium sulphate, ammonium chloride, ferrous sulphate, calcium chloride, manganese chloride
Max produ –calcium chloride, Enzyme production 74.5 ± 0.06 U / ml. Biomass 29.5 ± 0.01 mg / ml
Min produ- sodium nitrate, Enzyme production 38.4 ± 0.02 U / ml. Biomass 8.3 ± 0.02 mg / ml Physico-chemical Characterization of Samples
The collected tannery effluent sample was used to study parameters like pH and temperature.
Here the figure 2 shown the difference of pH and temperature in pretreated, P. aeruginosa sthc002AJ treated and keratinase treated sample. Figure 3 represent the chemical properties like
http://annalsofrscb.ro 1457 BOD, COD,TDS, chloride and hardness in order to determine the quality of raw textile effluent, bacterial strain treated effluent and extracellular enzyme effluent. The samples were analyzed according to the standard methods.
Fig 2: difference of pH and temperature in pretreated, P. aeruginosa sthc002 AJ treated and Keratinase treated sample
Fig 3: physico-chemical properties of raw textile effluent, bacterial strain treated effluent and extracellular enzyme effluent.
GCMS Analysis
0 5 10 15 20 25 30 35
Pre treated effluentTreated with P. aeruginosa strain sthc002Keratiase treated
pH / Temperature ( ˚C )
pH Temperature
0 500 1000 1500 2000 2500
BOD COD TDS Chlorides Hardness chromium
Parameters ( mg / l )
Pre treated
Treated with P. geniculata AJ Protease treated
http://annalsofrscb.ro 1458 The list of identified compounds from both pretreated and treated sample details including name, retention time, molecular weight, formula and structure were given in table 1 and 2.
Table 1: GC-MS Analysis of Raw Tannery Effluent S.
No
Name of the
compound
Retenti on time ( Minites )
Area
٪
Molecul ar
weight ( g / mol )
Molecul ar formula
Molecular structure
1 Acetic acid 9.296 10.28 2
60 C2H4O2
2 Methanol, (methyl- one- azoxy)-, acetate (ester)
9.616 18.26 5
131 C5H9O3
N
3 2-
butanone, 3-hydroxy
9.726 10.71 6
88 C4H8O2
http://annalsofrscb.ro 1459 4
Silane,dim ethyl
15.473 7.773 60 C2H8SI
5 2-Hydroxy ethylhydraz ine
15.603 8.883 76 C2H8ON
2
6 methylene- 2b-
hydroxyme thyl-3,3- dimethyl- 4b-(3- methylbut- 2-enyl
26.743 7.510 222 C15H26O
7 spiro[andro st-5-ene- 17,1'- cyclobutan]
-2'-one, 3- hydroxy-, (3.beta., 17.beta.)
26.823 9.328 328 C22H32O
2
http://annalsofrscb.ro 1460 8 3-chloro-5-
cholestene
26.913 14.32 9
404 C27H45Cl
9 2,6,10- dodecatrien -1-ol, 3,7,11- trimethyl- 9-
(phenylsulf onyl)-, (e,e)
27.228 6.317 362 C21H30O
3S
10 5,8,11,14- eicosatetrae noic acid, methyl ester, (all- z)
27.333 6.597 318 C21H34O
2
Table 2: GC-MS Analysis of Treated Tannery Effluent S.No Name of the
compound
Retentio n time
Area ٪ Molecula r weight
Molecular formula
Molecular structure
1 hexane, 1-
propoxy
23.057 23.777 144 C9H20O
2 2,6,6-trimethyl- bicyclo[3.1.1]he pt-3-ylamine
23.827 24.378 153 C10H19N
http://annalsofrscb.ro 1461
3 7-
oxabicyclo[4.1.
0]heptane, 1- methyl-4-(2- methyloxiranyl)
25.998 16.044 168 C10H16O2
4 Diazoprogester one
26.463 19.613 338 C21H30N4
5 2-
Hydroxyethylhy drazine
15.603 16.183 76 C2H8ON2
5 1-
adamantanemet hylamine, .alpha.-methyl
12.692 11.584 179 C12H21N
6 3-Amino-2- 13.928 21.376 102 C3H6O2 N2
http://annalsofrscb.ro 1462 oxazolidinone
Fig 4: GC-MS Spectrum of Hexane, 1-propoxy(RT 23.757) Detected in Treated Tannery Effluent.
http://annalsofrscb.ro 1463 Fig 5: GC-MS Spectrum of 2,6,6-trimethyl-bicyclo[3.1.1]hept-3-ylamine (RT 23.378) Detected in Treated Tannery Effluent.
Fig 6: GC-MS Spectrum of 7-oxabicyclo[4.1.0]heptane, 1-methyl-4-(2-methyloxiranyl)- (RT 16.044) Detected in Treated Tannery Effluent.
http://annalsofrscb.ro 1464 Fig 7: GC-MS Spectrum of Diazoprogesterone (RT 19.613) Detected in Treated Tannery Effluent.
Fig 8: GC-MS Spectrum of 1-adamantanemethylamine, .alpha.-methyl (RT 12.69 ) Detected in Treated Tannery Effluent
http://annalsofrscb.ro 1465 Fig 9: GC-MS Spectrum of 3-Amino-2-oxazolidinone (RT 13.928) Detected in Treated Tannery Effluent
The above project work results that tannery effluent was grey colored, turbid with a disagreeable odor, acidic in pH, with high organic and inorganic load indicating high values of TDS, total hardness, calcium, magnesium, sodium chloride, sulphate, heavy metal and chromium. From the present findings, the level of turbidity (O.D), pH and Physico-chemical parameters found to decreases from high to moderate or optimum level. The most attractive feature of P. aeruginosa sthc002micro organism is very effective against tannery effluent and microbe treated water can be used for irrigation purpose. The study on this topic explains that the P. aeruginosa sthc002would play an important role in tannery effluent treatment. We check both pre treated and treated tannery effluent sample.
Saravanan. R et al. explained in his work the tannery samples shows similar results when it treated with Bacillus subtilis (MTCC 1427).In another work Sivaprakasam et al donebacteria which are tolerant to salt were isolated from marine and tannery saline wastewater samples were discovered as P.aeruginosa, B. flexus, E. homiense and S. aureus. Growth factors of the identified strains were optimized. They were identified salt inhibition effects on chemical oxygen demand removal rate and comparative analysis was made by treating the tannery effluent with activated sludge obtained from CETP. Natural environmental microorganisms present in raw tannery saline wastewater .
Compare with other studies in our study we done the GCMS for the treated and pretreated tannery effluent sample so that we can find out the presented compounds, by using these results we can easily identify the quantity and presence of different compounds. From figure 4 to 9, it represent the GC-MS spectrum of treated tannery effluent. From the figure 3 you can understand the amount of chromium was very low in pretreated and treated (Keratinase and P. aeruginosa
http://annalsofrscb.ro 1466 sthc002AJ) samples, similarly you can see the TDS value was higher in pretreated sample. In the case of temperature and pH was almost similar in treated and pretreated samples.
Bioremediation has been used as a strategy to remove pollutants. It may be due to the concurrent process of absorption and metabolism properties of the microbial consortia, includes the heavy metal tolerance by permeability barrier, intra-cellular and extra-cellular sequestration, active transport efflux pumps, enzymatic methods and also reduction in the sensitivity of targeted cellular organelles to metal ions It is evident that Keratinase and P. aeruginosa sthc002AJ are very effective against tannery effluent. Based on these results the enzyme and microbes treatment method can be applied for large scale tannery effluents treatment.
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Published 2008 Apr 29.
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