Citation: Kumar R, Kumar R, Chauhan A, Kumar M, Goyal MK, et al. (2015) Characterization and Isolation of Fungi for Removal of Color from Pulp DQG3DSHU0LOO(IÀXHQW0HHUXW ,QGLD -(QYLURQ$QDO7R[LFRO GRL 10.4172/2161-0525.1000324
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was used for the continuous enrichment of the fungal strain has been used at the rate of 24 gm per liter of distilled water. The pH of the media was adjusted to 5.0 ± 1.0. Minimal Salt Medium (MSM): The sterilized minimal salt medium was used for the continuous enrichment of the microbial strains in this investigation. The composition is described above. Antibiotic: Antibiotic streto penicillin was used for the prevention of any contamination of the media used in the investigation. It has been used at the rate of 100 mg per liter sterilized media. Isolation of fungus from the sediments: 1.0 gm of sediments was dissolved in 10 ml of autoclaved water. It was shaken vigorously to mix them properly. It was kept standing at room temperature for 2 hr. Then the supernatant was decanted and centrifuged at 1000 rpm for 5 minute. The same process was repeated with the sediments of both the industry. Serial dilution of the supernatant in the order of 10 -3 , 10 -4 , 10 -5 were done using 0.1 ml autoclaved double distilled water from each dilution was spread on the PDA plats and incubated at 30°C for 4 days. Spreading was done in triplicate. Preparation of fungal inoculums (Pellets): For the preparation of fungal inoculums in the form of pellets initially the fungal isolates were individually grown on potato dextrose agar plates by incubation at 30°C for 4 days. Fungal mycelium disc (of about 1 cm diameter) were cut for the zone of active growth and inoculated at the rate of mycelia discs inn sterilized Erlenmeyer flasks containing potato dextrose broth (100 ml) and steriptopenicilliini (100 ml). The flasks were incubated at 30°C for 4 days under shake conditions in orbital shaker. The pellets of approximately 1.5-2.0 mm size were observed suspended in the medium ready for use in the treatment of pulp and paper mill effluent. Screening of potential strains MSM effluent (150 ml) in a sterilized Erlenmeyer flasks inoculated with individual fungal isolates and a control were then inoculate with individual fungal isolates and a control were then incubated at 30°C in a rotary shaker for 15 days. The parameter such as color and lignin were observed and measured at an interval of 0, 1, 3, 5, 7, 10, 15 days. The experiment was repeated on the basis of comparative analysis of reduction percentage of different parameters studied by the individual isolates along with control; the potential strains were screened out. Decolonizing assay Color was estimate by the method 212°C (APHA). The sample was filtered and then centrifuged at 10,000 rpm for 30 minutes to remove all the suspended matter. The supernatant was taken and volume of sample was maintained at 50 ml. The pH of the sample was adjusted to 7.0 with 2M NaOH. Then the absorbance was taken at the wavelength of 456 nm using Varian Cary Spectrophotometer. The value of color was calculated in coloring unit by making standard curve [3-8]. Preparation of standard curve Dissolved 1.246 gm of potassium chloroplatinate and 1.0 gm crystallized cobaltous chloride in ware with 100 ml core HCl and diluted to 1 liter. This stock solution has color of 500 CU. The suitable dilution was made from this stock solution to prepare standard curve. Measurement of Lignin The lignin of the samples was estimated according to Pearl and Benson. In this method the sample was centrifuged at 10,000 rpm for 30 minutes to remove all the suspended matter. The pH of the
Installed capacity (tons per year-TPY)
State
Number of mills
Andhra Pradesh
22
414550 220500
Assam
2 4
Bihar
25000
Gujarat Haryana
68 15
935800 149150
Himachal Pradesh Jammu and Kashmir
6 1
53200
5000
Karnataka
14
34530
Kerala
5
215600 290650 1034050
Madhya Pradesh
21 71
Maharashtra
Nagaland
1 1 1
33000 27050
Odissa
Pondicheri
9000
Punjab
37
375162
Rajasthan Tamil Nadu
7
12195
31 73 18
639250 870780 222600
Uttar pradesh West Bengal
Total 6121327 Table 1: Geographical spread of the pulp and paper mill in India. 406
The rest mills are small and have a production capacity of less than 30 tons per day of over 300 mills producing about 2.5 million tons of pulp about 10 percent is pulp for viscous rayon grade industry. The pollution potential of small paper mills is greater than that of large mills as they lack proper effluent treatment system and chemical recovery units. Characteristics of pulp and paper mill effluent The paper making processes produces an effluent which contains a substantial quality of cellulose “fines” and others additives. This can be up to 50% of the total mass. This contaminated water is frequently referred to as “white water”. Reclamation of the effluent is economically essential as the gross usage of water in the industry is very high and cost of effluent treatment for all water assigned to drain would be too expensive and would also involve a loss of raw materials [6-13]. Materials and Methods Sampling Sampling was started for the purpose of isolation of fungal strains. The sediment was collected from the Kaccha nala outside the Centaury Pulp and Paper Mill (Ghanshyam Dham, Lalkuan, Nainital and Uttaranchal) premises. Second sampling was done near Anand Tissue Paper Mill, Meerut. Sediments samples were collected from the drain near the industry. The Sediment was collected in clean plastic containers and brought to the laboratory of the department and immediately stored in refrigerator at 4°C unit used for further analysis. Microbial community procurement Culture media: The composition of the culture media which was used in the present investigation has been given as under: Potato Dextrose Agar (PDA): The sterilized potato dextrose agar media used for culturing the fungal strains was used at the rate of 39 gm per liter of distilled water and the pH of the media was adjusted to 5.0 ± 1.0. Potato Dextrose Broth (PDB): The sterilized potato dextrose broth
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