proforma for submission of project proposals on research...
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PROFORMA – I
PROFORMA FOR SUBMISSION OF PROJECT PROPOSALS ON RESEARCH AND DEVELOPMENT, PROGRAMME SUPPORT
(To be filled by the applicant)
PART I: GENERAL INFORMATION
1. Name of the Institute/ Project Proposal : Dr. N.G.P. Arts and science college , Coimbatore
2. State : Tamilnadu
3. Status of the Institute : College affiliated to Bharathiar University, Coimbatore
4. Name and designation of the Executive Authority of the Institute/University forwarding the application : Dr. P.R. Muthuswamy, Principal Dr. N.G.P. Arts and Science College, Coimbatore
5. Project Title : Screening, Optimization and Molecular Characterization of High potential Dye Degrading Indigenous Microbes from Noyyal
River Basin, Tamilnadu.
6. Category of the Project (Please tick) : R&D
7. Specific Area (Please see Annexure - II) : 3. Environment (Microbiology) 3.1. Bioremediation
8. Duration : 3 Years
9. Total Cost (Rs.) : Rs. 37, 61,000 (Thirty seven lakhs and sixty one thousand rupees)
10. Is the project Single Institutional or Multiple-Institutional (S/M) ? : Single institutional 11. If the project is multi-institutional, please furnish the following: NA 12. Scope of application indicating anticipated product and processes
• Our research focus is to substitute physical and chemical mode of effluent treatment by biological remediation. • This study will give the clear knowledge and characteristics of native organisms
from noyyal river, tamilnau and their dye degrading potential. • It will be cost effective and eco – friendly.
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13. Project Summary (Not to exceed one page. Please use separate sheet) : Enclosed (ANNEXURE- I)
PART II: PARTICULARS OF INVESTIGATORS Principal Investigator : 14. Name : Dr.S.S. Sudha
Date of Birth : 01/05/1972
Sex (M/F) : Female
Designation : Professor& Head
Department : Microbiology
Institute/University : Dr. N.G.P. Arts and Science College (Affiliated to Bharathiar University)
Address : Dr. N.G.P- Kalapatti Road, Coimbatote, Tamilnadu.
PIN: 641048
Telephone : 0422- 2369221, Mob : 09245282174
Fax : 0422-2369206
E-mail : [email protected]
Number of Research
projects being handled at present : Nil
Co-Investigator 15. Name : Mr. S. Senthil Prabhu
Date of Birth : 24/07/1980
Sex (M/F) : Male
Designation : Assistant Professor
Department : Microbiology
Institute/University : Dr. N.G.P. Arts and Science College (Affiliated to Bharathiar University)
Address : Dr. N.G.P- Kalapatti Road, Coimbatote, Tamilnadu.
PIN : 64 1048
Telephone : 0422- 2369221, Mob : 09842814320
Fax : 0422-2369206
E-mail : [email protected]
Number of Research projects being handled at present: Nil
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PART III : TECHNICAL DETAILS OF PROJECT
16. Introduction
Screening, Optimization and Molecular Characterization of High potential Dye Degrading Indigenous Microbes from Noyyal River Basin, Tamilnadu
India's dye industry produces every type of dyes and pigments close to 80,000 tones.
India is the second largest exporter of dyestuffs and intermediates (developing countries) after
China. The textile industry accounts for the largest consumption of dyestuffs; at nearly 80%.
Industrialization is vital to a nation’s economy because it serves as a vehicle for development.
However, there are associated problems (pollutions and adverse effect) resulting from the
introduction of industrial waste products into the environment.
Textile dyes are chemicals with complex aromatic structures designed to resist the effects
of laundering and sunshine, for example a great number of dyes and other chemicals are used in
textile wet processing. There are more than 105 commercially available dyes with over 1×106 tons
of dyestuff produced annually world-wide. Dyes are a synthetic origin and complex aromatic
molecular structures, which make them stable and difficult to degrade.
The textile industries utilize large volumes of water in its processing operations and
generate substantial quantities of wastewater. Dye wastes are usually discarded into water with or
without processing. Aesthetic merit, gas solubility and water transparencies are affected by the
presence of dyes even in small amounts. The presence of colour in waste water is very important
and usually contributes the major fraction of biochemical oxygen demand. Many classes of
synthetic dyes used in the textile and dyeing industries play a predominant role in almost every
type of application.
The textile factories daily discharge millions of litres of untreated effluents in the forms of
wastewater into public drains that eventually empty into rivers (Olayinka and Alo, 2004). This
alters the pH, increases the biochemical oxygen demand (BOD) and chemical oxygen demand
(COD), and gives the rivers intense colorations. The use of these water resources is limited and
the ecosystem is affected. Colored industrial effluent from the dyeing industries is one more
reason for pollution of aquatic systems which leads major environmental problems due to its
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impact on water bodies and growing public concern over its toxicity and it spoil the agriculture
land too.
Bioremediation using a variety of microbes for the degradation of xeno-biotics seems a
green solution to the problem of environmental pollution. Microbes have been gifted by nature
with the ability of degrading a wide spectrum of environmental pollutants. Microorganisms are
the most abundant organisms in natural freshwater systems and also play a key role in ecological
processes. Detailed knowledge of the diversity and function of microorganisms dwelling in
freshwater habitats is an essential prerequisite for the sustainable management of freshwater
resources. Bacteria, filamentous fungi and microalgae have been extensively evaluated for the
decoloration and degradation of dyes and colored effluents.
Microbes and their enzymes have the potentials to degrade complex and organic
compounds into simpler fragments; sometimes achieving complete mineralization. There has been
an intensive research on microbial decolorization of dye waste waters. Moreover, filamentous
fungi’s extracellular enzymes where Oxidative enzymes (Ferreira et al, 2006) and Bacteria’s
thermostable azoreductase (Maier et al, 2004) plays a significant role in biodegradation of Dyes.
Algae are photosynthetic organisms, which distributed in nearly all parts of the world and in all
kinds of habitats. Alga can degrade number of dyes, postulating that the reduction appears to be
related to the molecular structure of dyes and the species of algae used (Yan and Pan, 2004).
Compared with chemical and Physical methods, biological processes have received more
interest because of their cost effectiveness, lower sludge production and environmental
friendliness. Bioremediation is a cheaper alternative for colour removal in textile effluents.
Decolourization of dye wastewater by microbial metabolic activities and the mixed culture of the
microbes are the subject of many studies which is also our extension but through indigenous
microbial population, a rare prospect also.
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16.1 Origin of the research Proposal
The major and minor rivers flowing in Tamilnadu were the major source for surface and
ground water. One of such rivers flowing in Coimbatore, Erode and Karur districts is the Noyyal
River. The river Noyyal is a seasonal river and it originates from Vellingiri hills in the Western
Ghats of Coimbatore district. It flows through Coimbatore, Erode and Karur districts and finally
joins into Cauvery River near Noyyal village. It flows over a distance of 175 kilometers. The
catchment area of the river is 3.49 lakh hectares. Throughout its distance on both sides of the
banks of river Noyyal, more than 100 villages are situated. Noyyal was the major source for
irrigation, drinking water and other activities of the people living on both sides of the river and
even for people living beyond 3 kms from the river.
The industrial effluents released by dyeing and bleaching factories in Tirupur, a major
hosiery centre in South India has become a serious issue because, it has severe impact on water
bodies. After semi-treatment or without treatment, the effluents are released into Noyyal river. At
present there are about 750 dyeing and bleaching industries in Tirupur. The effluents released by
these units are stored in Orathupalayam Dam, which was constructed in 1991 at the cost of
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Rs. 1646 lakhs. The water-spread area is 1049 acres and it was expected to irrigate 500 acres in
Erode district and 9875 acres in Karur district.
The effluent has altered the nature and quality of both soil and water, which subsequently
and indirectly affect the health and the progeny of human beings. Environmental changes are
inevitable, but at what rate the change is happening will have an impact over the dwellings of
earth because, the drift should provide enough time to adapt, which is not available in the current
problem. This has motivated us to go for indigenous micro- flora that will be a divine force in
striving for welfare of both environment and economy.
Dying industry and garments are purely related to chemistry and Textiles, where the
survival of these firms and its employees has become a big question mark due to the legal and
responsible interference by environmental activists, farmers and guidelines by department of
environment and forestry, government of India. It is essential that everything survives and
environment is protected without fail. Our research purely aims at understanding micro-biota of
Noyyal river basin and discovering the possibilities for biological remediation; it creates a
significant role in installing the knowledge of microbiology for renaissance of both dying and
garment industries.
16.2 (a) Rationale of the study supported by cited literature
Dyes may also significantly affect photosynthetic activity in aquatic life by reducing light
penetration intensity and may also be toxic to some aquatic fauna and flora due to the presence of
aromatics, metals, chlorides, etc. (Dhaneshvar et al., 2007).
Dyes are resistant towards conventional methods of waste water treatment (Joshi et
al.,2004).
The economic and safe removal of the polluting dyes is still an important issue. Because
all the physicochemical methods are very expensive and results in the production of large amount
of sludge, which creates the secondary level of land pollution. In this situation bioremediation is
becoming important, because it is cost effective and environmental friendly and produces less
sludge (Chen et al.,2003).
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Biological process have potential to convert or degrade the pollutant into water, carbon
dioxide and various salts of inorganic nature. The isolation of potent species and there by
degradation is one of the interest in biological aspect of effluents treatment (Mohan et al., 2002).
Therefore, the use of microbial communities for on-site treatment of dye containing waste
waters from textile and dye-stuff industries could be an economical alternative (Ola et al., 2010).
Development of efficient dye degradation requires a suitable strain and its use under
favorable conditions to realize the degradation potential. In recent years there has been an
intensive research on fungal decolorization of dye wastewater. It is turning into a promising
alternative to replace or supplement present treatment processes (Mohandass Ramya et al., 2007).
Most of the Microorganisms have detoxifying abilities (that is, mineralization,
transformation and or immobilization of pollutants), microorganisms, particularly bacteria, play a
crucial role in biogeochemical cycles for sustainable development of the biosphere (Tropel and
Van der Meer, 2004).
Many microorganisms belonging to different taxonomic group of bacteria, fungi,
actinomycetes and algae have been reported for their ability to decolourize azo dyes. Many
bacteria are capable to degrade azo reactive dyes aerobically and anaerobically (Kuhad et al.,
2004).
The presence of extra bands in TLC in decolourized samples indicates the ability of the
bacterium to degrade dyes enzymatically and can be exploited for bioremediation of azo dyes and
their derivates containing wastes, since it seem to have the potential to degrade the toxic azo dyes
into non-toxic product form (Usman Aftab et al., 2011).
Decolorization of textile dyes from fungi Aspergillus Flavus, Fusarium oxysporum,
F.moniliforme and Trichoderma harzianum isolated from the soil samples around the textile
distillery industries of Nanjangud, Karnataka, India were tested their efficacy in decolorization of
textile dyes, Orange 3R, Blue 3R, Yellow GR etc. It was found that all the four fungal species
were found to be efficient in decolouring in textile dyes (Raju et al., 2007).
The potential of Cosmarium species, belonging to green algae, was investigated as a
viable biomaterial for biological treatment oftriphenylmethane dye, Malachite Green (MG). The
results obtained from the batch experiments revealed the ability of algal species in removing dye.
The effects of operational parameters (temperature, pH, dye concentration and algal
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concentration) on decolorization were examined. Optimal initial pH was determined 9 (Daneshvar
et al., 2006).
b) Hypothesis
I) This study is to thorough investigation of high potential dye degrading indigenous microbes
from noyyal river basin .
H1: The indigenous microbes has the high efficiency for dye degradation when individual
Strain.
H2: The consortium of indigenous microbes has high efficiency to dye degradation.
H ∅ : The indigenous microbes of noyyal river basin doesn’t have dye degradation efficiency.
II) This study also investigates the enzymes produced by isolated microbes on effective dye
degradation.
H1 : The enzymes from indigenous microbes posses high efficiency in dye degradation
H ∅ : The enzymes from indigenous microbes are not efficient in dye degradation.
C) Key questions
1. Why this proposed research is important?
2 How many bacterial strains are having high efficiency on dye degradation from
noyyal river basin, Tamilnadu?
3. How many fungal strains are having high efficiency on dye degradation from
noyyal river basin ?
4. How many microalgae can effectively degrade the dyes from noyyal river basin?
5. At what extent mixed cultures (consortium) of native microbes will degrade the dyes?
6. Will the enzymes from native microbes has the high potential to dye degradation?
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16.5 Current status of research and development International status
Wastewater effluents from the textile and other dye-stuff industries contain
significant amounts of synthetic dyes that require treatment to prevent groundwater
contamination. In research aimed at biotechnology for treatment of azo dyes, this study examined
4 strains of azo-dye degrading bacteria to identify efficient strains and determine incubation times
required for decolorization (Mohammad Reza et al.,2011).
The potential of Trametes villosa and Pycnoporus sanguineus to decolorize reactive textile
dyes used for cotton manufacturing in the State of Minas Gerais, Brazil, was evaluated. Growth
and decolorization halos were determined on malt extract agar containing 0.002g L-1 of the
dye(Katia et al., 2006).
Textile effluent-adapted bacteria were isolated from effluent samples collected from the
discharge and drainage pipes of three textile industries in the Oshodi/Isolo Local Government
area of Lagos, Nigeria. The textile effluent non-adapted bacteria were isolated from soil samples
taken from a municipal landfill. As a preliminary step in the development of textile effluent
biotreatment processes involving indigenous microbes, we have discovered textile effluent
adapted strains of Acinetobacter and Bacillus, and effluent non-adapted Bacillus species with
potential use in effluent treatment. (Olukanni et al., 2006).
Practical development of bacteria for bioaugmentation requires careful screening that is
based not only on their efficacy in pure culture, but also on their ability to compete with the
indigenous microbial communities in wastewater streams and ability to be produced and delivered
as a stable inoculum. (Azeem Khalid et al.,2010).
Activated sludge samples collected from wastewater treatment plant (WWTP) in textile
factory “Giorgetti Bulgaria” AD were exploited for isolation of dye decolorizing bacteria. A
microbial strain AZO29 was selected based on its efficiency, showing maximum and faster
decolourization of textile dye Amaranth. Identification of isolate by 16SrRNA technique revealed
that the microorganism were from family Pseudomonadaceae of Gammaproteobacteria (99%
confidence), the closest matches found after BLAST alignment were 16S sequences of
Pseudomonas strains (97% similarity) (Grekova-Vasileva et al., 2009).
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The industrial manufacturing of azo dyes and textile finishing processes generate
wastewater streams contaminated with azo dyes. Some of the azo dyes are difficult to treat by
conventional wastewater treatment methods. Compared with physical and chemical methods,
biological techniques are preferable because of economical advantages and low possibility of by
products production. Screening potential microorganisms is a critical step in the construction of
an effective remediation system (Usman Aftab et al., 2011).
Biological processes have potential to convert or degrade the pollutant in water into
carbon dioxide and various salts of inorganic nature. (Hazrat Ali et al., 2009).
Biological methods of treatment combined with physical or chemical methods or both for
colour removal are immensely useful and cost effective. It is therefore imperative to study the
metabolic and enzymatic systems of fungi in order to gain insight into their degradation
metabolisms. Amongst the fungi studied so far, wood degrading white-rot fungi has been reported
to have the potential to successfully treat coloured industrial effluents (Moturi et al.,2009).
In recent years, the ability of microorganisms to decolorize textile wastewater has received
great attention due to the environmental persistence and toxicity of these pollutants. In this paper
biological decolorization of triphenylmethane dye,C.I. Basic Green 4 (BG 4), by Chlorella
species was investigated. The effect of operational parameters (temperature, pH, initial dye
concentration and algal concentration) on decolorization efficiency was examined (Kahtee et
al.,2009).
National Status
Biological decolourization has been investigated as a method to transform, degrade or
mineralize azo dyes. In the present studies bacteria from soil from dye waste area were subjected
for acclimatization to C.I. Reactive yellow 145, an azo dye in the basal nutrient media.The most
promising bacterial isolate was used for further dye degradation studies. The 16s r RNA gene
sequencing revealed the isolated organism as Enterococcus faecalis strain YZ66. A preliminary
identification of the culture was based upon biochemical tests. 16sr-RNA sequencing of the
isolated organism was done in GeneOmbio Technologies Pvt. Ltd., Pune, India. Phylogenetic tree
was constructed by the Neighbour –Joining method (Sahasrabudhe and Pathade et al., 2011).
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A number of biotechnological approaches have been suggested by recent research as of
potential interest in combating this pollution source in an eco- efficient manner. The ligninolytic
enzymes of white-rot fungi (e.g., Mn peroxidases, lignin peroxidases, and laccases,) are directly
involved not only in the degradation of lignin in their natural lignocellulosic substrates, but also in
the degradation of various xenobiotic compounds including dyes (Indira Priyadarshini et
al.,2011).
The Pleurotus sajor caju silver nanoparticle effectively decolorized the dye within
24hours of incubation when compared with its plain culture (Pleurotus sajor caju) which takes
more than 48 hours for the same process.This is for the first time reporting that Pleurotus sajor
caju silver nanoparticle was used for the decolorization of the dye Congo red (Nithya and
Ragunathan,2011).
Isolation and identification of dye decolourizing fungal isolates from textile dye effluent
was carried out. The isolates of Aspergillus flavus, Aspergillus niger, Helminthosporium sp.,
Mucor sp. and Penicillium sp. were isolated from the textile effluent samples collected from
Elampillai Jalagandapur, Tamilnadu . Different parameters were used for optimizing. Conditions
for maximum decolourization effect by the fungal isolate (Ponraj et al.,2011).
The study describes degradation ability of A.tamaraii, mixed fungal culture (Trichoderma
sp. and A.flavus) and Penicillium purpurogenum on various synthetic dyes.A.tamaraii decolorized
>90% of Coomassie brilliant blue, Phenol blue, and malachite green dyes (Ramalingam et al.,
2010).
Decolourisation of textile dyes from fungi Aspergillus flavus, Fusarium oxysporum,
Fusarium moniliforme and Trichoderma harzianum isolated from the soil samples around the
textile distillery industries of Nanjangud, Karnataka, India were tested for their efficacy in
decolourisation of textile dyes (Raju et al., 2007).
Frequently encountered thirteen fungi isolated from dye contaminate soils were assessed
for their decolourisation and biodegradation of three sulphonated azo dyes. The fungal
organisms showed 41-80% decolourisation of scarlet red and 80-90% of fast greenish blue and
48-89 % of brilliant violet. For the decolourisation the responsible enzymes, K-amylase, protease
and catalase were estimated. A. niger showed maximum secretion of K-amylase in 14 days of
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incubation. The protease production was ranged from 25-38 U/ml by majority of the fungi tribe
T. notatum was found to be efficient in production of catalase. This study is the fungal based
treatment system for the reclamation of the dye contaminated soils (Laxminarayana, 2010).
Development of efficient dye degradation requires a suitable strain and its use under
favourable conditions to realize the degradation potential. In recent years there has been an
intensive research on fungal decolorization of dye wastewater. It is turning into a promising
alternative to replace or supplement present treatment processes (Mohandass Ramya et al., 2007).
16.6 The relevance and expected outcome of the proposed study
This study is to provide the high potential dye degrading organisms from noyyal river basin and
molecular characterization is to give clear knowledge of high efficiency dye degrading Indigenous
organisms. So this study will give the solution for major environmental problem & solution for dyeing
industries with cost effective and eco friendly method.
16.7 Preliminary work done so far
Studies on decolorization of textile dyes by white rot fungi- Trametes hirsute (2010).
The Study studies on white rot fungi Trametes hirsute given the greatest extent in
decolorization of Blue RR, Blue FNG, and Black dyes. Various parameters have been optimized
and dye degradation is analysed by GCMS. Blue RR was decolorized at 35o C, and at pH 4
using maltose as a carbon source and Yeast extract as nitrogen source. Blue FNG was
decolorized at 30o C and pH 3 using maltose as carbon source and yeast extract as nitrogen
source. Black dye was effectively decolorized at 35o C at pH 4 using glucose as carbon source
and yeast extract as nitrogen source. The laccase enzyme production from above fungi were
well optimized of for different parameters. Decolorization of dyes with Immobilized beads of
crude enzyme (laccase) extract was also observed and dye degradation was analysed by
GCMS. Along with other fungi like Aspergillus fumigates, Penicillium sp. and Fusarium
oxysporum were also used for degradation of various dyes.
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17. Objectives
Ø Collection of samples from noyyal river basin and industrial effluents. Ø Isolation and Screening of dye degrading potential organisms. Ø Identification of Bacteria, fungi and microalgae. Ø Molecular based Characterization of Isolates. Ø Optimization of culture condition and enzymes for effective dye degradation.
Ø Effect of mixed culture (consortium) on biodegradation. Ø Toxicity studies on plants, animals and microbes. Ø The field trial will be conducted using the enzymes and cell biomass.
18. Work Plan (Year-wise Plan of work and targets to be achieved)
18.1 Methodology 1st year 1.1Collection of sample
Soil, water samples from noyyal river basin, effluents from industries will be collected and
samples are transport to the laboratory.
1.2 Isolation of Organisms
Bacterial isolation by serial dilution on nutrient agar and maintain on same medium. Soil
Fungi will be isolated by serial dilution and plating on PDA (potato Dextrose Agar) medium.
Microalgae isolation and it will be subcultured on growth (MN) medium. (Khataee et al., 2009).
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2nd Year 2.1 Screening of organisms Screening of the bacterial isolates in Bushnell and Hass medium amended with different
concentrations of dyes, (Prachi et al., 2009). The fungal isolates to be screened for dye
degrading activity (Subramanyan, 1971). The dye degrading efficiency for isolated microalgae
will be checked by growth medium ( Incubation in glass jar with growth medium t 25 ο C for 15
days and exposed in light) amended with dye (Khataee et al., 2009) .
2.2 Decolorization assay
Decolorization activity will be analysed by UV-VIS Spectrophotometer (Moorthi et al.,
2007, Nehra et al., 2008).
2.3 Identification of Screened Organisms
The identification of bacterial isolates by gram staining, morphology, biochemical
methods and other standard microbiological methods (Cappuccino and Sherman, 1999). The
identification of fungal taxa will be made based on the Manual of Soil Fungi (Gilman, 1957);
2002, Rafi M, and Sjjad – Ur-Rahman, 2002). Hyphomycetes (Subramanyan, 1971); Dematiceous
and More Dematiceous Hyphomycetes (Ellis, 1971); Identification of cyanobacteria by using the
standard taxonomic manuals of Geitler (1932) and Desikachary (1959).
2.4 Molecular characterization
16S rRNA gene sequencing, plasmid profiling will be done based on standard protocol.
(Azcarate-Peril and Raya,2001).
2.5 Enzyme assay The enzymes from isolates to be assayed by using UV spectrophotometer. (Hatvani and Mecs, 2001, Shanmugam et al.,1999, Moturi and Singara charya, 2009).
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2.6 Optimization Optimization of physicochemical characters like temperature, pH, Carbon sources,
Nitrogen sources for potential dye degradation by the cultures and enzymes (Rajendran et al.
2011, Ponraj et al., 2011).
2.7 Estimation of protein by lowry’s method. 2.8 Design and development of Microbial consortium & immobilization Various combinations of isolated native microbes to developing consortium by applying
Cayley’s table (Rajendran et al.,2011). Immobilized cells also will be used to analysis the dye
degradation potential (Ohmomo et al. 1987; Shin et al. 2002).
2.9 Biodegradation activity analysis After completion of degradation, the decolorized medium will be analyzed in HPLC (High
performance liquid chromatography), FTIR (Fourier Transform Infrared Spectroscopy) UV-VIS
Spectrophotometer (Faisal et al.,2006).
3rd Year 3.1 Toxicity studies Phytotoxic studies on plants, Microbial toxicity by zone of inhibition method and toxicity
of both treated and untreated dyes will be checked on various animals and aquatic organisms
(Puwaneswari et al.,2006; Subhasini Sharma et al.,2007).
3.2 Field trails
The high potential dye degrading organisms, Mixed cultures, and enzymes will be applied
in bioreactors, industrial effluent treatment plants and the contaminated sites.
3.3 Report Preparation The results obtained will be interpreted by statistical analysis, finally the report preparation with documental evidences.
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18.2 Connectivity of the participating institutions and investigators (in case of multi- institutional projects only) : Not Applicable 18. 3. Alternate strategies (if the proposed experimental design or method does not work what is the alternate strategy) : Molecular cloning (recombinant technology) will be applied to isolated organisms for effective degradation of dyes. 19. Timelines: (Please provide quantifiable outputs) Period of study Achievable targets 0-6 Months Collection of review
6-12 Months Collection of samples and Isolation
12-18 Months Screening , identification and Molecular characterization
18-24 Months Optimization by different parameters for enzymes and cell biomass for dye degradation
24-30 Months Toxicity studies, Effect of consortium on dye degradation
30-36 Months Field trial (application) and report preparation
20. Name and address of 5 experts in the field
Sr.No. Name Designation Address 1. Dr. R. Rajendran Associate
professor Department of Microbiology, PSG College of Arts and Science College, Coimbatore- 14.Tamilnadu.
2. Dr.S. Rajakumar Assistant professor
Department of Marine Biotechnology Bharathidasan University, Trichirapalli-24, Tamilnadu
3. Dr.P.M. Ayyasamy Assistant professor
Department of Microbiology, Periyar University Salem-11, Tamilnadu
4. Dr.P..Lakshmanaperumalsamy Registrar Karpagam University, Coimbatore-21. Tamilnadu
5. Dr. K. Selvam Head & professor
Department of Biotechnology, Dr.N.G.P.Arts & Scince College, Coimbatore- 48, Tamilnadu
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Budget (In Rupees)
PART IV: BUDGET PARTICULARS
A. Non-Recurring (e.g. equipments, accessories, etc.)
S. No. Item Year 1 Year 2 Year 3 Total 1. Phase contrast microscope 1,50,000 1,50,000 2. HPLC 8,00,000 8,00,000 3. Gradient PCR 4,00,000 4,00,000 4 Books & Journals 1,20,000 1,20,000
Sub-Total (A) = 14,70,000 B. Recurring B.1 Manpower (See guidelines at Annexure-III)
S. No. Position No.
Consolidated Emolument
Year 1 Year 2 Year 3 Total
1. Project fellow Rs.12,000 /m 1-2 Yrs Rs. 14,000/m 3rd yer
1,44,000 1,44,000 1,68,000 4,56,000
Sub-Total (B.1) =4,56,000 B.2 Consumables
S. No.
Item Qty Year 1 Year 2 Year 3 Total
1. Chemicals, Reagents - 90,000 1,50,000 80,000 3,20,000 2. Glass wares - 75,000 75,000 60,000 2,10,000 3. Molecular characterization
kit - - 2,00,000 - 2,00,000
4. Animals for toxicity Studies
- 1,00,000 1,00,000
Sub-Total (B.2) =8,30,000
Other items Consolidated Emolument
Year 1 Year 2 Year 3 Total
B.3 Travel & field work
1,00,000 75,000 1,00,000 2,75,000 B.4 Contingency & hiring - 75,000 75,000 80,000 2,30,000 B.5 Overhead
- 5,00,000 Sub-total of B (B.1+B.2+B.3+B.4+B.5)
22,91,000
Grand Total (A + B) 37,61,000
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PART V : EXISTING FACILITIES Resources and additional information
1. Laboratory:
a. Manpower Lab assistants: 2 (no’s)
b. Equipments
S.No Name of the Instrument
1. Autoclave vertical (350 X 550 mm)
2. UV – VIS Spectrophotometer
3. Electrophoresis apparatus
4. High Speed Centrifuge
5. Laminar air flow chamber
6. Incubator
7. BOD Incubator
8. Rotary Evaporator
9. Ultra Cooling Centrifuge
10. Shaker
11. Colorimeter
12. UV Transilluminator
13. pH Meter
14. Magnetic Stirrer
15. Hot plate
16. Incubatory Shaker
17. Deep freezer
18. Clinical centrifuge
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19. Hot air oven
20. Qubec colony counter
21. Membrane filter unit with vaccum pump
22. Soxhlet apparatus
2. Other Infrastructural facilities:
Animal House, Fungal Room, Incubation room, Research Laboratory, Sterilization and Decontamination rooms.
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PART VII: PROFORMA FOR BIOGRAPHICAL SKETCH OF INVESTIGATORS Provide the following information for the key personnel in the order listed on PART II. Follow
this format for each person. Principal Investigator
Name : Dr. S.S. Sudha Designation : Professor & Head
Department/Institute/University : Microbiology/ Dr. N.G.P. Arts & Science College/ Bharathiar
University
Date of Birth: 01/05/1972 Sex (M/F); Female (39yrs) SC/ST: NA
Education (Post-Graduation onwards & Professional Career)
Sl No. Institution Place
Degree Awarded
Year Field of Study
1. Kandasamy Kandar College, Paramathy Vellur
M.Sc 1999 Microbiology
2. AVVM Sri Pushpam College, Thanjavur
PhD 2006 Microbiology
3. Bharathiar University, Coimbatore
MBA 2010 Hospital Management
A. Position and Honors Position and Employment (Starting with the most recent employment)
Sl No. Institution Place
Position From (Date) To (date)
1. Dr.N.G.P. Arts & Science College, Coimbatore
Professor & Head 01/06/2006 Till date
2. Shrimathi Indira Gandhi College, Trichy
Asst. Professor 01/06/2004 20/05/2005
3. Kurinji college of arts and Science, Trichy
Asst. Professor 01.06.1999 30.05.2004
Honors/Awards : 1. Acted as chair person for the technical session in International Conference on
Bioresource Technology organized by Department of Botany Nirmala college for Women on Antihyperlipidemic activity of spirulina species in diet induced albino rats. During 7 & 8th October 2010.
2. Acted as chair person for judging projects presented on National Children’s
Science Congress by Coimbatore district school students at GRD College on
12th November 08.
3. Invited speaker in Coimbatore Management Association Weekly Meeting on
24.11.2008 and delivered a speech on “Disaster Management. Professional Experience and Training relevant to the Project : NA
22
B. Publications (Numbers only) : 35 Books: NA Research Papers, Reports: 35 General articles:..... Patents: NA Others (Please specify) NA Selected peer-reviewed publications (Ten best publications in chronological order)
1. S.S.Sudha and Parvathy A. “A study on Screening for L-Asparaginase Producing
Microorganism from Beach soil” Research J of Biol. Sci.: II .1, 2010 : 50-54.
2. Uma P., and S S Sudha “Biodecolorization of textile Reactive Dyes using Trametes
hirsute”. Research J of Biol. Sci. Vol : I No.1, 2009: 30-38.
3 S S Sudha. “Marine Environment: A Potential Source for L-Asparaginase Producing Microorganism” The Icfai University Journal of Biotechnology. Vol : 3 No.4, 2009 :57-62.
4.S.Kalaivani, A.MahalakshmiPriya, S.S.Sudha and S.Balasubramaniyan
Phycoremediation of sewage using Microalgae Chlorella sp. Nature
Environment and Pollution Technology.Vol 8, No1, 2009: 187-192.
5. Vimalin J, S.S. Sudha, D. Padmanabhan, A. Asarudeen, P. Ayyappan. Isolation &
Characterization of community acquired Methicillin Resistant
Staphylococcus aureus in school going children. Res J Biol Sci.
2009;1(3): 153-56.
6. R. Vijayakumar, S.S. Sudha, A. Pannerselvam, T Thaajuddin. A comparative
study on cultivation and nutritional status of Oyster Mushroom Pleurotus
ostreatus on different substrates. J.Sci.Trans.Environ.Technov, 2008.
1(4):209-212.
7. S.S.Sudha,A.Panneerselvam and N.Tajuddin “Antagonistic interaction of some
Mangrove Soil Fungi against plant pathogens-Curuvlaria lunata and
Drechslera ellisii” (J.Sci. Trans.Environ.Technov., 1(3):154-157, 2008.
8. S.S.Sudha,A.Panneerselvam and N.Tajuddin “Seasonal variation of cyanobacteria at
Muthupet mangrove environs, Tamilnadu,South India.( Seaweed Res.Utiln.,29
(1&2): 263-271,2007).
23
9. S.S.Sudha,A.Panneerselvam and N.Tajuddin “Antagonistic Potentials of
Mangrove Soil Fungi against plant pathogens” Indian Journal of Applied
Microbiology, 6(1): 61-65,2006).
10. S.S.Sudha,A.Panneerselvam and N.Tajuddin.“Antagonistic effect of some mangal
soil fungi against Colletotrichum falcatum went. and Curvelaria pallescens
(Walker) Boedijn” .Geobios32, 288-290,2005 .
11. S.S.Sudha,A.Panneerselvam and N.Tajuddin .“Antagonistic Potential of some
Mangrove Soil Fungi against Fusarium oxysporum and Rhizoctonia solani”
J.Microb. World 7(2) PP.140-143, 2005..
Publications relevant to the proposed area of work
Publications
Ø Uma P., and S S Sudha “Biodecolorization of textile Reactive Dyes using Trametes
hirsute”. Research Journal of Biological Science. Vol : I No.1, 2009: 30-38.
Ø S.S.Sudha and Parvathy A. “A study on Screening for L-Asparaginase Producing
Microorganism from Beach soil” Research Journal of Biological Science. Vol : II
No.1, 2010 : 50-54.
Ø S.S.Sudha,A.Panneerselvam and N.Tajuddin “Seasonal variation of cyanobacteria at
Muthupet mangrove environs, Tamilnadu,South India.( Seaweed Res.Utiln.,29 (1&2):
263-271,2007).
Ø S.S.Sudha,A.Panneerselvam and N.Tajuddin “Antagonistic interaction of some
Mangrove Soil Fungi against plant pathogens-Curuvlaria lunata and Drechslera
ellisii” (J.Sci. Trans.Environ.Technov., 1(3):154-157, 2008).
Ø R. Vijayakumar, S.S. Sudha, A. Pannerselvam, T Thaajuddin. A comparative study
on cultivation and nutritional status of Oyster Mushroom Pleurotus ostreatus on
different substrates. J.Sci.Trans.Environ.Technov, 2008. 1(4):209-212.
24
Ø S.Kalaivani, A.MahalakshmiPriya, S.S.Sudha and S.Balasubramaniyan
Phycoremediation of sewage using Microalgae Chlorella sp. Nature Environment and
Pollution Technology.Vol 8, No1, 2009: 187-192.
Biographical sketch of Co- Investigator
Name : Mr. S. Senthil Prabhu Designation : Assistant Professor
Department/Institute/University : Microbiology/ Dr. N.G.P. Arts & Science College/ Bharathiar
University
Date of Birth: 24/07/1980 Sex (M/F) ; Male (30yrs) SC/ST: NA
Education (Post-Graduation onwards & Professional Career)
Sl No. Institution Place
Degree Awarded
Year Field of Study
1. Periar University, Salem M.phil 2008 Microbiology
2. G.R.Damodaran College of Science, coimbatore
M.Sc 2005 Microbiology
3. Dr.N.G.P. arts and science college, coimbatore
B.Sc 2001 Biochemistry
A. Position and Honors Position and Employment (Starting with the most recent employment)
Sl No. Institution Place
Position From (Date) To (date)
1. Dr.N.G.P. Arts & Science College, Coimbatore
Assistant Professor 01/06/2008 Till date
2. Nehru arts and science college, Coimbatore
Asst. Professor 01/06/2006 30/04/2008
Honors/Awards : Professional Experience and Training relevant to the Project : NA
B. Publications (Numbers only) : 06 Books: NA Research Papers, Reports: 05 General articles:01 Patents: NA Others (Please specify) NA
25
26
References.
Ajibola VO, Oniye SJ, Olubodi T, Umeh UG ,2005. Biodegradation of Indigo containing textile effluent using some strains of bacteria. J.Appl. Sci., 5(5): 853-855. Azcarate-Peril MA, Raya RR (2001) Methods for plasmid and genomic DNA isolation from lactobacilli : Food microbiology protocols, methods in Biotechnology, J.F.T. Spencer A.L. Ragout de Spencer. Humana press INC., Totowa, New Jersey. Azeem Khalid, Muhammad Arshad, and David Crowley,2010. Bioaugmentation of Azo Dyes, Hdb Env Chem 9: 1–37. Cappuccio JG and N.Sherman,1999. Microbiology: A laboratory Manual. 4th edition , Addison Wesley Longman, Inc., Harlow, England, 199-204. Chen KC, Wu JY, Liou DJ, Hwang SCJ.2003. Decolorization of the textile azo dyes by newly isolated bacterial strains. J Biotechnol 101; 57-68. Dhaneshvar, N., Ayazloo, M., Khatae A.R., Pourhassan M., 2007. Biological decolourization of dye solution containing malachite green by microalgae Cosmarium sp. Bioresource Technology 29, 1-7. Ferreira, V. S., Magalhaes, D. B., KIing, S. H., De Silva, J. G. &Bon, E. P. 2006. N- emethylation of methylene blue by ligninperoxidase from Phanerochaete chrysosporium. toichiometricrelation for H2O2 consumption. Applied Biochemistry and Biotechnology 84: 255-265. Ganesh Parshetti, Satish Kalme, Ganesh Saratale, and Sanjay Govindwar,2006. Biodegradation of Malachite Green by Kocuria rosea MTCC 1532, Acta Chim. Slov., 53, 492–498. Grekova-Vasileva M, I. Popov, D. Vassilev, Y. Topalova, 2009. Isolation and characterisation of microbial strain azo29 Capable of azo dye decolourization, biotechnol. & biotechnol. Eq. 23/2009/se xi anniversary scientific conference. Hazrat Ali, Wesal Ahmad and Taqweemul Haq.2009. Decolorization and degradation of malachite green by Aspergillus flavus and Alternaria solani. African Journal of Biotechnology Vol. 8 (8):1574-1576. Indira Priyadarshini R., V. Bhuvaneswari and K. Suresh Kumar .2011. Isolation Identification a nd Phylogenetic analysis of white rot fungus and Heterologous Expression of Gene Encoding Laccase. Journal of Applied Sciences in Environmental Sanitation. Vol (6): 69 - 8 3. Joshi M, Bansal R & Purwar R, Colour removal from textile effluents, Indian J fiber Textile Res, 29 : 239-259. Kátia M.G. Machado; Luciana C. A. Compart; Rúbio O. Morais; Luiz H. Rosa; Mércia H. Santos, 2006. Biodegradation of reactive textile dyes by basidiomycetous fungi from brazilian ecosystems, Brazilian Journal of Microbiology 37:481-487.
27
Khataee A. R., Pourhassan M. & Ayazloo M. 2009,Biological Decolorization of C.I. Basi c Green 4 Solution by Chlorella sp.: Effect of Operational Param eters, Chin J Appl Environ Biol , 15 ( 1 ): 110~114. Klich, MA,2002. Identification of common Aspergillus sp. 1st edition. Centralbureau voor Schimmelcultures, Urecht, Netherlands, 122. Laxminarayana E, M. Thirumala Chary, M. Randheer Kumar and M. A. Singara Charya, 2010.Decolourisation and biodegradation of sulphonated azo dyes by fungi to clean dye contaminated soil environments, J Nat Env Sci , 1(1):35-42. Sahasrabudhe M. M. and G. R. Pathade,2011. Decolourization of C.I. reactive yellow 145 by Enterococcus faecalisstrain YZ66, Archives of Applied Science Research, 3 (3):403- 414. Maier J, Kandelbauer A, Erlacher A, Cavaco – Paulo A, Gubits GM, 2004. A new alkali – thermostable azoreductase from bacillus sp.Strain SF. Appl. Environ. Microbial., 70: 837- 844. Mohammad Reza Majid Khoshkholgh Pahlaviani, Alireza Massiha, Khosro Issazadeh, 2011.An Innovative Approach to Biodegradation of Textile Azo Dyes by native bacterial strains in Iran, 2011 International Conference on Biotechnology and Environment ManagementIPCBEE vol.18 (2011) © IACSIT Press, Singapoore. Mohan S.V, Roa C.N, Prasad K.K, karthikeyan J., 2002. Treatment of simulated reactive yellow 22 (Azo) dye effluents using spirogyra species. Waste manage. 22, 575-582. Mohandass Ramya, Bhaskar Anusha, S. Kalavathy, and S. Devilaksmi1.2007. Biodecolorization and biodegradation of Reactive Blueby Aspergillus sp. African Journal of Biotechnology Vol. 6 (12): 1441-1445. Moorthi, SP, Selvam, PS, Sasikalaveni, Murugesan AK. & Kalaichelvan, PT. (2007). Decolorization of textile dyes and their effluents using white rot fungi. African J. Biotechnol. 6 424-429. Moturi, b and Singara charya, M. A.2009, Decolourisation of crystal violet and malachite green by fungi, Science World Journal Vol 4 :28-3. Naeem Ali, 2006. Biotechnological Approaches for the Treatment of Textile Dyes by Indigenous Microorganisms, Phd Thesis (Department of Microbiology/ Quaid-i- Azam University of Islamabad). Nehra, K. Meenakshi. A. & Malik. K. (2008). Isolation and optimization of conditions for maximum decolourization by textile dyedecolourizing bacteria. Poll. Res. 27 257- 264. Nithya.R and Ragunathan.R, Decolorization of the dye congored by Pleurotus sajor caju silverNanoparticle, 2011 International Conference on Food Engineering and BiotechnologyIPCBEE vol.9 (2011) © (2011)IACSIT Press, Singapoore.
28
Ohmomo, S., Kaneko, Y., Sirianuntapiboon, S., Somchai, P., Atthasampunna, P., Nakamura, I.1987. Decolorization of molasses waste water by a thermophilic strain, Aspergillus fumigatus G2-6. Agric. Biol. Chem. 51, 3339-3346. Olukanni O. D., Osuntoki, A. A. and Gbenle, G. O, 2006. Textile effluent biodegradation potentials of textile effluent-adapted and non-adapted bacteria, African Journal of Biotechnology Vol. 5 (20), pp. 1980-1984. Ponraj M , P. Jamunarani , V. Zambare,2011. Isolation and Optimization of Culture Conditions For Decolorization of True Blue Using Dye Decolorizing Fungi, Asian j. Exp. Biol. Sci. Vol 2(2): 270-277. Prachi Kaushik and Anushree Malik, 2009. Microbial decolourization of textile dyes through isolates obtained from contaminated sites, Journal of scientific & Industrial Research, 68:325-331. Puvaneswari N, J muthukrishnan and P Gunasekaran, 2006. Toxicity assessment and microbial degradation of azo dyes, India J. of Eper. Biol., 44:618-626. Rafi M, and Sjjad – Ur-Rahman, 2002. Isolation and identification of pseries. Int.J. Agri. Biol., 4: 552-557. Rajendran R, karthik sundaram and K Uma maheswari, Aerobic Biodecolorization of mixture of azo dye containing effluent using adapted microbial strains, J Env Sci & Tech,2011. Raju, N. S., Venkataramana, G. V., Girish, S. T., Raghavendra, V.B. & Shivashankar, P. 200). Isolation and evaluation of indigenous soil fungi for decolourization of textile dyes. Journal ofApplied Sciences 7: 298-301. Ramalingam, N.Sarawathy, S.Shanmugapriya, S.Shakthipriyadarshini, S.Sadasivam and M Shanmugaprakash, 2010. Decolorization of textile dyes by Aspergillus tamaraii, mixed fungal culture and Penicillium purpurogenum, Journal of Scientific & Industrial Research 69:151-153. Sathiya moorthi, P., Periyar selvam, S., Sasikalaveni, A. Murugesan, K., and Kalaichelvan, P. T. 2007.Decolorization of textile dyes and their effluents using white rot fungi, African Journal of Biotechnology Vol. 6 (4):. 424-429. Shanmugam, V., Kumari, M. and Yadav, K. D. (1999).n-proponol as substrate for assaying the lignin peroxidase activity of Phanerochaete chrysoporium.Indian Journal of Biochemistry and Biophyscis 36: 39 – 43 . Shazia Erum and Safia Ahmed 2011. Comparison of dye decolorization efficiencies of indigenous fungal isolates. African Journal of Biotechnology Vol. 10(17) :399-3411. Shin, M., Nguyen, T. and Ramsay, J. 2002. Evaluation of support materials for the surface immobilization and decolorization of Amaranth by Trametes versicolor. Appl. Microbiol.Biotechnol. 60, 218-223.
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Subhasini Sharma, Kalpana, Arti, Shweta, V. Suryavathi, Pawan K. Singh, Ramesh, Shipra and K.P. Sharma,2007 Toxicity assessment of textile dye wastewater using swiss albino rats, Australasian J of ecotoxicology Vol. 13, pp. 81-85, 2007. Tropel D, Van der Meer JR (2004). Bacterial Transcriptional Regulators for Degradation Pathways of Aromatic Compounds. Microbiol. Mol. Biol. Rev. 68: 474-500.
Usman Aftab, Muhammad Riaz Khan, Muhammad Mahfooz, Musadiq Ali, Salik Hassan Aslam and A. Rehman, 2011. Decolourization and Degradation of Textile Azo Dyes by Corynebacterium sp. Isolated From Industrial Effluent. Pakistan J. Zool., vol. 43(1), 1-8. Yan, H., Pan, G., 2004. Increase in biodegradation of dimethyl phthalate by Closterium lunula using inorganic carbon.Chemosphere 55, 1281–1285.
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Annexure - I
Project Summary
This study is to isolate bacteria, fungi and microalgae by various microbiological
methods. The isolated organisms are screened for dye degradation potential. Screened
organisms are subjected to molecular characterization, 16s rRNA sequencing after
polymerase chain reaction (PCR) and this to construct the phylogenetic tree, plasmid
profiling of screened organisms also will be done. The Enzymes from isolates will be used
for determination of potential dye degradation, this enzymes will be estimated by Lowry et al
method. Optimization of enzymes and cell mass by different physiological and biochemical
parameters for efficient dye degradation. The immobilized cells will be used to determine the
dye degradation potential. Toxicity studies will be conducted on plants, microbes and
animals. Finally above things will be practically applied to the fields as well as bioreactors in
Tirupur dying industries, and contaminated sites of noyyal river paves, its a way for natural
remediation for environment and remedy for dyeing industries.