MYCOREMIDIATION OF

CONGO RED - AN AZO DYE

MANGAI.G,
II M.Sc., Biotechnology,
Guided By: Mr.SOURAV BHATTACHARYA, M.Sc.,
Faculty of Microbiology,
Genohelix,
Jain University,
Bangalore 560019
 INTRODUCTION
 Environmental degradation due to rapid
growing population and economic
development.
 Chemical wastes and byproducts or
discharges of poorly-treated or
untreated sewage from the dye and
textile industries are the major source of
water pollution.
 Bioremediation agents in the treatment of
wastewater containing textile dyes, which
are the major source of aromatic amines.
 More than 10,000 dyes used in textile
industry and 280,000 tons of textile dyes
are discharged every year worldwide.
 Azo dyes contribute to about 70% of all
used dyes, and have complex structure
and synthetic nature.
 The colour of dyes is due to azo bond and
Nitrogen to nitrogen double bond (–N N–)
and associated chromophores.
 Azo compounds are xenobiotic compounds
that are very recalcitrant against
biodegradative processes.
 Disposal of dyes into surface water not only
affects the aesthetic but cause also
biotoxicity.
 Congo red is one of the predominantly used
azo dye in the textile industries. It is a
secondary diazo dye and toxic in nature.
 Many physicochemical techniques are high
cost, low efficiency and inapplicability to a
wide variety of dyes.
 A definitive solution of the colour problem of
textile effluents would provide a marked
competitive advantage for this industrial
sector.
 OBJECTIVES
♦ Screening of the laboratory fungal
forms for degradation activity.

♦ To check the degradation activity

under static and shaking condition.

♦ To check the degradation activity when

dye is the sole source of Nitrogen.
♦ To check the degradation activity
when media is free of CuSO4.

♦ To check the degradation activity

when media is supplemented with
inorganic fertilizer.

♦ To check the degradation activity

when media is supplemented with
heavy metals.
METHODOLOGY
 Screening of the laboratory fungal
forms for degradation activity.

• Plating of semi synthetic media with congo

red (0.050g/L).

• Plates are inoculated with the fungal species such

as Aspergillus niger, Aspergillus oryzae,
Aspergillus flavus, Penicillium spp, Cladosporium
spp, Pleurotus spp.

• Zone of decolourization around the fungal growth

was observed after the incubation at 27° C.
 To check the degradation activity
under static and shaking condition.

• Preparation of Semi synthetic broth with congo

red (0.050g/L).

• Inoculation of fungal species.

• Incubation at static and shaking condition (120

rpm).

• Decolourization observed through

spectrophotometer at 570 nm.
 To check the degradation activity when
dye is the sole source of Nitrogen.

• Preparation of Semi synthetic broth with

congo red (0.050g/L) as the sole Nitrogen
source.

• Inoculation of fungal species.

• Incubation at static and shaking condition

(120 rpm).

• Decolourization observed through

spectrophotometer at 570 nm.
To check the degradation activity when
media is free of CuSO4.

• Preparation of Semi synthetic broth without

CuSO4 and with congo red (0.050g/L).

• Inoculation of fungal species.

• Incubation at static and shaking condition (120

rpm).

• Decolourization observed through

spectrophotometer at 570 nm.
 To check the degradation activity
when media is supplemented with
inorganic fertilizer (NPK fertilizer).

• Preparation of Semi synthetic broth with 1%

NPK fertilizer and congo red (0.050g/L).

• Inoculation of fungal species.

• Incubation at static and shaking condition (120

rpm).

• Decolourization observed through

spectrophotometer at 570 nm.
To check the degradation activity when
media is supplemented with heavy
metals.

• Preparation of Semi synthetic broth with

HgCl2 (0.1%) & congo red (0.050g/L).

• Inoculation of fungal species.

• Incubation at static and shaking condition

(120 rpm).

• Decolourization observed through

spectrophotometer at 570 nm.
 RESULTS
♦ Screening of the laboratory fungal forms for
degradation activity has shown the zone of
decolourization.
♦ The percentage of decolourization varied between 8.6%
- 32.5% (static) & 10.9% - 98.86% (shaker).
♦ When dye is used as the sole Nitrogen source, the
percentage of decolourization varied between 4.68% -
24.75% (static) & 8.41% - 39% (shaker).
♦ When SS broth is free of CuSO4, the percentage of
decolourization varied between 5.98% - 29.9% (static)
& 4.5% - 89.6% (shaker).
♦ When media is supplemented with NPK fertilizer the
percentage of decolourization varied between 47.82% -
81.2% (static) & 49% - 99.8% (shaker).
♦ When media is supplemented with the heavy metal HgCl 2
the percentage of decolourization varied between 89.1%
Decolourization activity is calculated by the formula
- 95.5% (static) & 88% - 90.6% (shaker).
Initial absorbance – Observed absorbance × 100
Initial absorbance
DECOLOURIZATION BY Aspergillus flavus

U – Uninoculated sample
A - 24 hrs
B - 48 hrs
C - 72 hrs
D – 96 hrs
 RESULTS
To check the degradation activity
under static and shaking condition.
Uninoculated sample’s OD @ 570 nm = 0.440
S.N O D @ 570 nm
MICRO 24 hrs 48 hrs 72 hrs 96 hrs
ORGANISMS
ST SH ST SH ST SH ST SH

1 Aspergillus niger 0.423 0.409 0.420 0.399 0.413 0.390 0.402 0.384
2 Aspergillus oryzae 0.401 0.397 0.386 0.386 0.367 0.385 0.355 0.225
3 Aspergillus flavus 0.399 0.336 0.390 0.273 0.354 0.005 0.297 0.005
4 Penicillium spp 0.410 0.410 0.405 0.389 0.400 0.405 0.398 0.376
5 Cladosporium spp 0.394 0.411 0.391 0.405 0.388 0.400 0.382 0.392
6 Pleurotus spp 0.396 0.402 0.392 0.398 0.389 0.392 0.381 0.364
 RESULTS
To check the degradation activity when dye
is the sole source of Nitrogen.
Uninoculated sample’s OD @ 570 nm = 0.832
S.N O D @ 570 nm
MICRO 24 hrs 48 hrs 72 hrs 96 hrs
ORGANISMS
ST SH ST SH ST SH ST SH

1 Aspergillus niger 0.794 0.784 0.790 0.739 0.776 0.724 0.775 0.709
2 Aspergillus oryzae 0.783 0.730 0.757 0.696 0.738 0.690 0.693 0.660
3 Aspergillus flavus 0.764 0.587 0.743 0.555 0.686 0.525 0.626 0.507
4 Penicillium spp 0.811 0.767 0.783 0.767 0.757 0.762 0.737 0.762
5 Cladosporium spp 0.802 0.771 0.799 0.769 0.795 0.766 0.793 0.761
6 Pleurotus spp 0.803 0.773 0.792 0.760 0.788 0.759 0.787 0.733
 RESULTS
To check the degradation activity when
media is free of CuSO4.
Uninoculated sample’s OD @ 570 nm = 0.618
S.N O D @ 570 nm
MICRO 24 hrs 48 hrs 72 hrs 96 hrs
ORGANISMS
ST SH ST SH ST SH ST SH

1 Aspergillus niger 0.614 0.596 0.589 0.596 0.587 0.596 0.581 0.590
2 Aspergillus oryzae 0.585 0.585 0.569 0.550 0.533 0.171 0.520 0.141
3 Aspergillus flavus 0.583 0.582 0.559 0.440 0.478 0.184 0.433 0.184
4 Penicillium spp 0.583 0.583 0.583 0.570 0.512 0.167 0.486 0.064
5 Cladosporium spp 0.583 0.582 0.581 0.582 0.574 0.503 0.569 0.440
6 Pleurotus spp 0.570 0.570 0.570 0.570 0.569 0.450 0.555 0.410
 RESULTS
To check the degradation activity when media
is supplemented with inorganic fertilizer(NPK
fertilizer).
Uninoculated sample’s OD @ 570 nm = 0.506
S.N O D @ 570 nm
MICRO 24 hrs 48 hrs 72 hrs 96 hrs
ORGANISMS
ST SH ST SH ST SH ST SH

1 Aspergillus niger 0.318 0.340 0.019 0.330 0.011 0.250 0.001 0.214
2 Aspergillus oryzae 0.267 0.295 0.034 0.240 0.005 0.205 0.005 0.205
3 Aspergillus flavus 0.225 0.302 0.009 0.241 0.016 0.142 0.001 0.095
4 Penicillium spp 0.299 0.310 0.295 0.300 0.264 0.266 0.264 0.264
5 Cladosporium spp 0.299 0.300 0.271 0.282 0.268 0.269 0.258 0.222
6 Pleurotus spp 0.295 0.307 0.272 0.285 0.244 0.285 0.243 0.265
 RESULTS
To check the degradation activity when media is
supplemented with heavy metal (HgCl2).
Uninoculated sample’s OD @ 570 nm = 1.785
S.N O D @ 570 nm
MICRO 24 hrs 48 hrs 72 hrs 96 hrs
ORGANISMS
ST SH ST SH ST SH ST SH

1 Aspergillus niger 0.238 0.238 0.219 0.217 0.185 0.198 0.176 0.199
2 Aspergillus oryzae 0.204 0.224 0.170 0.217 0.089 0.196 0.080 0.176
3 Aspergillus flavus 0.188 0.212 0.158 0.199 0.109 0.181 0.100 0.168
4 Penicillium spp 0.218 0.222 0.216 0.222 0.158 0.217 0.152 0.214
5 Cladosporium spp 0.217 0.214 0.191 0.201 0.167 0.187 0.163 0.167
6 Pleurotus spp 0.216 0.210 0.212 0.210 0.196 0.194 0.194 0.193
THANK ‘U’