INTRODUCTION:

A large number of chemical, petrochemical, fertilizers, paper, pharmaceuticals, leather processing, agro products industries are producing variety of organic as well as in organic chemicals. Some finished products of certain industries can be used as raw material for other industries so that variety of different chemicals is being formed for the satisfaction of human beings for luxurious, comfort and needs. In this process human beings are destroying the purity of the environment by adding different pollutants in atmosphere, on land and in water. Some of these pollutants are harmful to all the leaving organisms. Out of these pollutants water-solubles cause the pollution of water and affect the environmental balance. This insists the purification of wastewater and industrial effluent.

Even though environment gets polluted, development should not be halted, it must go hand in hand with environment. Therefore different activities in development should be such that they should have none or negligible adverse effect on environment.

Therefore during the development certain methods of controlling pollution must be employed. By these method total control of pollution is not possible but conversion of toxic and harmful pollutants to less harmful before flow out from industries must be deducted.

To minimize water pollution, number of methods have been given out of these biological method i.e. biodegradation of pollutant by microorganism is among the new technique in biotechnology to minimize water pollution. ¹

For degradation of artificial or manmade organic pollutant in water some microorganisms can be used. Number of scientist in other part of world have proved the use of microorganism in degradation of organic compounds, which are toxic and harmful to animal beings.

Degradation of all organic pollutants by single species of microorganisms is not possible but combination of species works together to degrade the number of organic pollutants. Environmental conditions, stability and adaptability of the species in the process of degradation is the important factor and it decides the success fo the process.

As the chemical industries producing different types of organic compounds and produces various wastewater streams. These streams differ in composition and strength of organic pollutants so they have different degree of degradability.

Also the domestic sewage water contains large number of pathogenic organisms including bacteria, protozoa, viruses etc. It also contains heavy metals organic and in organic compounds.

Therefore with increased discharge of industrial wastes into sewage system. Levels of heavy metals organic and inorganic toxic substances in municipal waste water and streams gets increased and water gets highly polluted. Therefore treatment of wastewater must be done with suitable method to purify water.

In the present research work attention is given to isolate suitable microorganisms and develop a method for biodegradation of organic pollutant of wastewater and industrial effluents to minimize water pollution with the help of micro-organisms

Materials and methods strains and culture:

Species of micro-organisms were collected from wood chips, stream water soil, domestic waste water and effluent from industries around Khopoli, Lonawala and Bhimashankar Forest. Some fungal forms in water growing parasitically and later on saprophytically on the algal forms. Each of the species collected was cultivated two times successively on Agar Media. Colonies showing good growth were grown up on yeast extract agar medium.²

The Micro-organisms were then cultivated individually with shaking in liquid medium.³ containing micro nutrient suitable Micro- organisms were selected by observing and studying their stability, capacity and degradability.

The pure strains of individual micro-organisms were isolated and some of them were maintained at 4^°C on malt extract agar and beef extract agar⁴

The activities of following various micro-organisms were studied for degradation.

A) Culture obtained from wood chips was :

1. Phellinus zeriensis

2. Streccherium hydneum

3. Coriolus versicolor

4. Fomes durissmus

5. Coniophora arida

B)Culture obtained from stream water, pounds, domestic waster water and industrial effluent were ⁵:

1. Chytridium lagenaria

2. Leptomitus lacteus (Roth)

3. Phylictochytrium quandricorne

4. Phlyctidium breviceps

5. Rhizophydium magrrhizum sparrow

6. Chlamydomonus sp

7. Chlorella sp

8. Spirulina sp

9. Oscillatoria sp

10. Bacteria (unknown)

All these micro-organisms studied with respect to stability and susceptibility and only few were found to be useful for waste water treatment for the present study of water purification and pollutant degradation following six micro-organisms were used

1 W1 Phellinus zeriensis

2 W2 Fomes durissmus

3 W3 Coriolus versicolor

4 W4 Streccherium hydneum

5 W5 Rhizophydium magrrhizum sparrow

6 W6 Mixed culture (Contains Rhizopus, Chlorella, Spirulina, Unknown Bacteria)

Collection of samples:

Water samples were collected from different part around Khopoli region, it includes mixed industrial effluents and domestic waste water. All samples were collected together on period of 10 days and representative sample were taken for studies. All these samples were collected and preserved as described by F.J. Welcher⁶. And analysed to find out the characteristic of wastewater and effluent used for the experiment within 4 hr. The characteristic of waste water used for experiments has been mentioned in table-1.

Table – I-Characteristics of wastewater used For the experiment

Sr. No.	Parameters	Characteristics
1	Acidity	10 mg/lit
2	Alkalinity	220 mg/lit
3	Total Dissolved solids	1250 mg/lit
4	Suspended solids	150 mg/lit
5	PH	8.62
6	Dissolved oxygen (D O)	1.85 mg/lit
7	Chemically dissolved oxygen (COD)	3600 ppm
8	Biological Oxygen demand (BOD)	95 mg/lit
9	Oils and greases	16 mg/lit
10	Phenolics	3 mg/lit
11	Chlorides	52 mg/lit

Biodegradation studies:

Experimental culture of W1, Phellinus zeriensis, W2 Fomes durissmus W3 Coriolus versicolor, W4 Streccherium hydneum were developed in a 20 ml liquid basal medium containing the following nutrient per liter of distilled water 0.2 gm, KH₂PO₄, 0.05 gm MgSO₄ 7H₂O, 0.01 gm CaC1₂, mineral solution, and vitamin solution 0.5 ml and pH maintained to 5.0 in 125 ml Erlenmeyer flasks for five days⁷

At the same time for culture W5 Rhizophydium magrrhizum sparrow, W6 Mixed culture (Contains Rhizopus, Chlorella, Spirulina, Unknown Bacteria) were developed in 20 ml liquid basal medium containing following nutrient per liter of distilled water 0.58 gm MgSO₄7H₂O, 67mg Cac1₂ 2H₂O, 2 mg NH₄M_OO₄, 4H₂O, 1gm (NH₄)² HOP₄, 3.5 gm KH₂PO₄, 4.5 gm K₂HPO₄per liter and pH was maintained to 7.0 in 125 ml Erlenmeyer flask for five days ⁴

Table - II Effect of microorganism on purification with respect to C O D in ppm and D O mg/lit After Seecific holding time

No	Micro organisms / No of Inoculation day --->	4th		5th		6th		7th
No	Micro organisms / No of Inoculation day --->	DO	COD	DO	COD	DO	COD	DO	COD
W1	Phellinus zerinsis	2	3350	3.1	1800	3.8	850	4.5	280
W2	Fomes durissmus	2	3210	3.1	1850	3.8	840	4.6	290
W3	Coriolus versicolor	2.1	3010	3.3	1750	4	700	5	240
W4	Streccherium hydneum	2.1	3025	3.3	1760	3.9	750	4.8	260
W5	Rhizophy dium magrrhizum sparrow	2.1	3200	3.1	1760	3.9	725	4.9	210
W6	Mixed culture (Rhizopus, chlorella, spirulina, unknown Bacteria)	2.3	2900	3.5	1650	4.2	690	5.2	170

Table – III- Effect of microorganism on waste water after 7 day treatment

No	Micro organisms	BOD gm/lit	Oil and Greases mg/lit	Phenolics mg/lit
W1	Phellinus zerinsis	20.5	2	0.1
W2	Fomes durissmus	21.8	1.9	0.15
W3	Coriolus versicolor	18.4	1.8	0.1
W4	Streccherium hydneum	20.3	1.9	0.1
W5	Rhizophydium magrrhizum sparrow	18.1	2.1	0.19
W6	Mixed culture (Rhizopus, chlorella, spirulina, unknown Bacteria)	17.2	2	0.19

For fermentation experiment 4 sets of wide mouth bottles of 2-liter capacity for each culture were used. 1000 ml of respective sample were taken in each bottle and neutralized with 0.2 M H2SO4 solution. For first 4 culture pH=5.0 and for last two culture pH=7.0 All the experimental bottles were fitted with rubber stopper through which two 6mm glass tubes were inserted. One tube extended into the bottle as gas inlet within the 1 cm of the sample fluid and outside it was connected to natural rubber tubing to a cotton plugged gas-sterilizing filter. The second glass tube as a gas outlet extended 1 cm or less into the bottle and outside it was connected to natural rubber tubing to cotton plugged as sterilizing filter^7,9,10

Bottles were flushed every day forcing air through the cotton filter at 100-200 ml per minute for 15 minutes

Then experimental bottles containing water sample were inoculated with five-day-old culture in respective bottle. Nutrient were added in the water sample in bottle after incubation of culture. Nitrogen was supplied from urea and phosphorous was supplied from diammonium phosphate to optimal ratio COD :N:P 200 : 5:1

Bottles were fitted with rubber stopper and aeration was done with fish tank aerator.

ANALYSIS:

After treatment of waste water sample with micro-organisms, treated water samples were analysed after 4,5,6,7 days for D.O. and COD while oil and greases, phenolics and B O D were analysed after 7 days ^6,8,11,12 and represented in Table II and III

RESULTS:

For white rot fungus W1 Phellinus Zeriensis, W2 Fomes durissmus W3 Coriolos versicolor, W4 Streccherium hydneum Were isolated from wood chips were used for water treatment. In the laboratory fermentation experiments were carried out at HP = 5.0 and the temperature 35 + 2^oC. Aeration was done with fish tank aerator. After inoculation of culture to water sample-holding time were chosen and analysis was done after 4^th , 5^th. 6^th and 7^th day and results are shown in Table – II and Table – III. The decrease in COD and increase in DO with increase in day of inoculation of different micro organism is shown in the figure – I

The analysis result shows that degree of water purification depends on the holding time of culture, with sufficient long residence time of culture in sample degradation of organic pollutant was more. The result indicate that decrease of COD after 4^th day of inocultation for culture W1, W2, W3. W4 was 6.94 to 16.38% and after 7^th day it was 91.94 to 93.33% while decrease of BOD after 7^th day was 77.05 to 80.2%.

The two micro organisms isolated from water stream and waste water W5 Rhizophydium magrrhizum sparrow, W6 Mixed culture (Contains Rhizopus, Chloralla, Spirolona, Unknown bacteria) were used more efficiently for water treatment. Their fermentation experiments were carried out at pH= 7.0 and temp 35+2^oc. Aeration was done with fish tank aerator. Analysis of treated waste water with W5 and W6 microorganism with same holding time show that the decreasing COD after 4^th day was 11.1 to 19.44% and after 7^th day was 94.16 to 95.27% while decrease of B O D after 7^th day was 80.94 to 81.89%

This indicates the microorganisms W5 and mixed culture of micro organism W6 were useful for waste water treatment.

DISCUSSION:

The result presented in this report shown that the several micro organisms were tested and four species of white rot fungi W1 Phellinus zeriensis, W2 Fomes durissmus W3 Coriolus versicolor, W4 Streccherium hydneum, one species of awuatic fungi W5 Rhizophydium magrrhizum sparrow, W6 Mixed culture (Contains Rhizopus, Chlorella, Spirulina, Unknown Bacteria) were used in this study. All there micro organisms were found to be competent in water treatment and degradation of organic pollutant with comparison of previous results⁽⁸⁾

From these fermentation experiments it is clear that it is essential to test the treatability of industrial wastewater at laboratory level. This will give the idea to what extent the degradation can be achieved by micro organisms. Then with selected microorganism further biodegradation studies can be done with different samples.

The different streams of wastewater and effluents were selected to study the degradation pattern. The representative sample of waste water was taken and analysed to find out its characteristic with respect to treatability. Domestic wastewater and industrial wastewater contain number of aliphatic and aromatic organic compound as pollutants so they show variable COD and BOD.

After treatment of wastewater with micro organism organic pollutant gates degraded and decrease in COD as well as BOD values were observed. They also increase the D.O. in the treated water sample. It helps to purity the wastewater.

This treated wastewater can be utilized for gardening. Floor washing, making lime solutions etc.

In the present work we have treated mixed sample of Domestic wastewater and industrial efficient with specific five micro organism and one mixed culture. Out of these one of the culture can be used for further detail studies on large scale water purification after deciding specific cultural parameters.

CONCLUSION:

Any wastewater or industrial effluent under study must be checked in laboratory for its treatability to get the good result after degradation.

Isolated purified cultures can be used as stock cultures and to obtain best result individual culture can be tested with waste water sample under specific cultural parameters like pH, temperature, atmosphere, medium etc.

Isolated culture can be used to decide mechanism by which the organic compounds were degraded and to find out the intermediates formed.

A suitable method for wastewater purification on large scale can be developed.

ACKNOWLEDGEMENT:

The authors are thankful to the management and the college authorities of Maharashtra Mahavidyalaya, Nilanga, K.M.C. College, Khopoli Dist Raigad and Rajarshi Shahu Mahavidyalaya, Latur for providing essential facilities for the present work, to Mr. Wakle G.V., Gadiwan N.K. for the computational work and also to Prof. T.G. Khamkar for maintaining the healthy ambience during the work.

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Received on 02.01.2013 Modified on 14.01.2013

Asian J. Research Chem. 6(2): February 2013; Page 106-110