AASCIT Journal of Biology 2015; 1(2): 15-24

Published online April 20, 2015 (http://www.aascit.org/journal/biology)

Keywords Waste Water Treatments,

Bioremediation of Direct Green

Dye,

Biosorption,

RFLP and Sequencing of 18S,

ITS rRNA,

Fungi Identification

Received: March 29, 2015

Revised: April 11, 2015

Accepted: April 12, 2015

Biodiversity Among Dominant Fungi Involved in Water Production from Non-Traditional Water Resources

Wafaa M. Abd El-Rahim1, *

, Abdelaal Shamseldin2,

Fatma H. Abd El Zaher1, Hassan Moawad

1, Eman Refaat

3

1Department of Agriculture Microbiology, National Research Centre, Dokki, Cairo, Egypt 2Environmental Biotechnology Dept, Genetic Engineering and Biotechnology Research Institute,

City of Scientific Research and Technology Applications, Alex 3Department of Microbiology, Faculty of Science, Al-Azhar University, Girls' Branco, Egypt

Email address wafaa10m@hotmail.com (W. M. A. El-Rahim)

Citation Wafaa M. Abd El-Rahim, Abdelaal Shamseldin, Fatma H. Abd El Zaher, Hassan Moawad, Eman

Refaat. Biodiversity Among Dominant Fungi Involved in Water Production from Non-Traditional

Water Resources. AASCIT Journal of Biology. Vol. 1, No. 2, 2015, pp. 15-24.

Abstract Now a day it is very important to remove industrial pollutants with minimum cost. The

present work focused on the assessment of extracellular laccases produced by different

fugal strains and its potential for Direct Green (DG) bio-removal from textile effluent

water, which can be used as a resource of non-traditional water. Ten fungal strains from

polluted sites were screened to test their capability of DG bioremoval to clean up waste

water containing-dye. Fungal dye removal capacity was assessed by measuring the

residual of DG dye in supernatant using spectrophotometer, over production of fungal

biomass and reduction of COD values after 28h of incubation. Results showed that the

fungal isolates could remove the simulated effluent containing-DG dye and they reduced

the COD values indicating on bio-removal of dye from the growth media. Strain FRD 11

was the most effective strain which removed about 85% of (300 mgl-1

) dye in 28h.

Molecular characterization of fungal isolates was done by using the restriction fragment

length polymorphisms (RFLP) and sequencing of ITS rRNA region as a reproducible

molecular method for fungal species identification and differentiation. RFLP of 16S

rRNA with CfoI proved the suitability of this technique for strain typing, species

identification and could divide them into three genetic clusters. Cluster 1 included strains

(FRD1, 2, 3 and 5), cluster 2 (FRD 4, 7, 8, 10 and 11) and cluster 3 (FRD 9).

Phylogenetic tree of ITS rRNA of five representative strains showed that strains FRD 7

and 11 were identified as Aspergillus tubigenesis, strain FRD 9 was identified as

Aspergillus niger and strains FRD1 and 3 were belong to Aspergillus terreus. These

results indicated that these fascinating new fungal strains can play significant role in

bioremediation and detoxification of effluents polluted with DG dye due to their ability

to biosorb or produce laccase enzyme-remediating dye.

1. Introduction

Industrial pollutants are the major environmental threats. Untreated industrial effluents

discharged into ecosystems cause serious problems to the surrounding ecosystems. One

of the greatest pollutants generated from industrial activities is the textile organic

pollutants. Textile effluents contain carcinogenic aromatic amines, dyes, organic and

inorganic materials (Ramachandran et al., 2013). Removing of textile dye from effluent

gained tremendous concern by environmentalists dye to the negative impact on the

16 Wafaa M. Abd El-Rahim et al.: Biodiversity Among Dominant Fungi Involved in Water Production from

Non-Traditional Water Resources

environment. Treatment of effluents polluted with textile dye

by physical and chemical methods is highly expensive, while

the use of biological process (bioremediation) using

microorganisms can eventually convert organic pollutants

into water and carbon dioxide. This process is much cheaper

and reliable method (Nehra et al., 2008; Wafaa and Moawad,

2010).

Many studies used bacteria for treating dye containing

effluents. Recently several other authors (Azmiet al., 1998;

Coulibaly et al., 2003; Wafaa et al., 2003b; Braret al., 2006;

Quet al., 2012; Tan et al., 2013) noted the suitability of fungi

as biological agents for removing and cleaning environments

from pollution of textile dyes. The use of fungal strains for

treating dye containing effluents was much advantageous than

the use of bacteria since fungi can remove dye residues by two

distinct mechanisms: biosorption and bioremediation (Tan et

al., 2013). This study focused on the degradation of widely

used direct green textile dye by fungal isolates to better

understand the bioremoval through biosorption and

bioremediation of the dye residues to reduce the environmental

threats. The biosorption is defined as binding of the dyes on

the surface of fungal cells (Crini, 2006; Crini and Badot, 2008).

This processes doesn't require energy, therefore, both dead and

living cells of fungi can accomplish the biosorption process.

Dyes bioremediation is defined as the breakdown of dyes

into carbon dioxide and water by specific enzymes. Through

this biological process dyes can be removed from aquatic

media without harming the environment. It is reported that

certain extracellular enzymes such as peroxidases and

phenoloxidases can degrade the dyes to carbon dioxide and

water (Duran et al., 2002).

This study directed for isolation and selection of fungal

strains highly efficient in removing textile DG residues dye

through biosorption and/or bioremediation. The study also

focused on the molecular differentiation of the fungal isolates

to identify the fungal species involved in bioremediation

process.

2. Materials and Methods

2.1. Dyes

Commercially used Direct Green dye was obtained from

Ixmadye (Dyestuffs and Chemicals Co.). All the chemicals

used were of analytical grade and a Dye stock solution

prepared and kept at -20°C until for further analysis.

2.2. Effluent Source

Effluents were collected from waste water produced from

industrial companies in Egypt: El-Mahala El-Kubra, Kafer

El-Dawar, New Borg El-Arab and Shubra El-Khima located

at four governorates; Gharbia, El- Behiera, Alexandria, and

Cairo. The effluents were collected in airtight plastic

containers. Samples were filtered to remove large suspended

particles and stored at 4°C until use. The textile waste

effluents were used for fungal isolation by dilution plate

method on a specific media. Fungal strains were grouped

based on their morphological characteristics. The

representative fungal isolates covering all morphological

variations were cultured and maintained on potato dextrose

agar medium (PDA) amended with direct violet dye at room

temperature. The culture was consistently sub-cultured every

15 days.

2.3. Isolation of Fungal Isolates from

Industry Effluents

Fungal isolates were obtained from dye effluent collected

from the previously mentioned sites. The effluent samples

were serially diluted seven times in tenfold (1/10) in sterile

water. Aliquots of diluted effluent samples (100 µl) from last

dilution were spread on agar plates and incubated at 280C for

5 days. Individual fungal colonies growing after this period

were re-streaked on potato dextrose medium (PDA) plates

containing Direct violet dye (300 mg l-1

) as carbon source,

and incubated at 28oC for 3 days then isolates were purified

by re-streaking on PD agar plates several times and checked

for their purity. Representative fungal isolates (10 isolates)

were selected based on high dye removing activity as clear

zone on the PDA plates. The purified isolates were examined

microscopically for their morphology to test their purity and

stored on PD media of agar slants. The selected fungal

isolates were designated as FRD (Fungal remediating dye)

and maintained on nutrient agar slants.

2.4. Preparation of Fungal Biomass Inocula

One square cm fungal mycelium (4 to 5 days old culture)

grown on ager plates was transferred to 250 ml of conical

flask containing PD broth medium and incubated on rotary

incubator shaker at 150 rpm and 28oC for 4 to 5 days to

obtain enough fungal biomass. Ten ml of the activated

growth were cultivated in 250 ml Erlenmeyer flasks

containing 100 ml of basal mineral medium supplemented

with 10gl-1

sucrose and 0.5g l-1

yeast extract. Flasks were

shaken on an incubator shaker (150 rpm) at 28oC for 4-5 days.

Fungal growth was separated by centrifugation at 8000 rpm

under aseptic conditions and the pellets were re-suspended in

sterile dye solution (500 mg l-1

). After re-suspension of

fungal growth, homogenization of suspension was made

using sterile magnetic bars on magnetic stirrer. Five ml were

withdrawn by manual sterile pipette and dried at 105°C to

determine the fungal biomass. Based on back calculations a

volume of fungal biomass containing 200-mg dry weight was

used as inoculums.

2.5. Determination of Dye Bioremoval

Capacity

Fungal removing capacity was measured by the difference

in dye absorption before and after incubation at specific time

intervals. Percentage of dye removal was calculated from

absorption values obtained against the controls using the

following equation:

AASCIT Journal of Biology 2015; 1(2): 15-24 17

Ei EfPercentage of Removing= 100

Ei

− ×

Where, Ei is the initial absorbance by dye solution and Ef

is the final absorbance of fungal growth media amended with

dye at different incubation time intervals.

2.6. UV- VIS Spectrophotometric Analysis

The synthetic dyes and diluted effluents containing dye

were analyzed using LBK spectrophotometer modal 4054 to

measure direct green dye concentrations at λ max 396 nm.

2.7. Dry Weight of Pellets and COD

Determination

Dry weight of pellets was obtained by filtering cultures

through filter paper and drying to constant weight at 65°C. At

the end of the experiment COD was measured as the oxygen

equivalent for the organic material in water. COD of the cell

free broth was measured at the end of the incubation period

by using a Hatc spectrophotometer test kit (HACH, CO)

adopting manufacture instructions.

2.8. Assay of Laccase (Lac) Enzyme

Lac activity was determined according to the method

described by Paszczynskiet al. (1988) which depend on the

oxidation of dimethoxy phenolic compound (DMP) by lac

enzyme. Six hundred µl of samples were mixed with 250 µl

of 250 mM sodium malonate buffer (pH 4.5) and 50 µl of 20

mM DMP then kept on water bath at 30ºC for 2 min. The

reaction was stopped by putting on ice for 10 min. The

enzyme activity was assessed by measuring the absorbance at

468 nm using 3UV vis spectrophotometer.

2.9. Molecular Differentiation and

Identification of the Fungal Strains

Fungal total DNA was extracted adopting the procedure of

Promega kit as manufacturer recommendations. Pure DNA

was concentrated and adjusted to give final concentration of

50 ng ml-1

. Template DNA was used to amplify a part of ITS

rRNA using the forward primer F 5-

TCCGTAGGTGAACCTGCGG-3, and reverse primer R 5-

TCCTCCGCTTATTGATATGC-3 (Whiteet al., 1990). The

standard PCR protocols applied with annealing temperature

at 52°C. The PCR product was examined by running on 0.8%

agarose gel incorporated with ethidium bromide and 100 bp

ladders for comparison. The fragment of ITS rRNA purified

using Qiagen kit and digested with CfoI and MspI overnight

and samples were run on 2% agarose gels for 20 min. RFLP

fragments were normalized using 100 bp ladders. The

purified ITS rRNA fragments were sequenced with the same

primers which were previously used for amplification based

on the Sanger di-deoxy method. Sequences were compared

with the available sequences from the Gene bank using NCBI

server and our sequences were deposited on the Gene Bank

under accession numbers KF289940-KF289943. Multiple

nucleotide sequence alignments were generated and edited

using ClustalW, as implemented in BioEdit. The ITS rRNA

multiple sequence alignment was manually adjusted to fit

that produced by the Ribosomal Database Project-II. Model

fitting was performed by likelihood ratio tests (LRTs) as

implemented in DAMBE. A phylogenetic tree was

constructed using a neighbor joining (NJ) phylogeny inferred

with the model selected by LRTs using MEGA2.1 (Kumar et

al., 2001) and the complete gap deletion option. The

robustness of the phylogeny was assessed by non-parametric

bootstrapping with 1000 pseudo replicates. Sequences of ITS

rRNA of five representative strains were constructed in a

phylogenetic tree for comparison with six sequences of the

Gene Bank. Sequences of the Gene Bank were from strains

Asperigillus terreus KAML04 (KC119260.1), Aspergillus

niger wxm76 (HM037958.1), Asperigillus tubigenesis cs/7/2/

(JN585941.1), Penicillium sp. vegaE2-22 (EF694630.1),

Trichoderma sp. DAOM222105 (Ay380901.1) and

Aseprigillus flavus strain DAOM225949 (JN938987.1).

3. Results and Discussion

3.1. Morphological Characteristics of Fungal

Isolates

Ten representative fungal isolates obtained from collected

waste water samples were examined to test their capacity to

remove green textile dye from aqueous media containing dye.

The fungal isolates were primary identified based on their

morphological properties to the genus level adopting the

standard fungal identification procedures (Gilman, 1957 and

Barnett and Hunter, 1972) Table (1). These 10 representative

isolates were selected to represent the morphological

variations among all fungal colonies on agar media used for

fungal isolation. These ten representative isolates likely

belonged to three species of genus Aspergillus.

Table (1). Effluent collection sites and morphological variations among representative isolates.

Isolate codes Governorate Sampling sites Genera Spore color

FRD 1, 2 and 3 Cairo Subra El-Khima Aspergillus, Brown

FRD 4 “ ‘’ ‘’ Gray

FRD 6 Alexandria New Borg El-Arab ‘’ Green

FRD 7 Cairo Subra El-Khima ‘’ Black

FRD8 and 9 El-Behiera Kafr El- Dawar ‘’ Black

FRD10 and 11 Gharbia El-Mahalla

El-Kubra ‘’ Black

18 Wafaa M. Abd El-Rahim et al.: Biodiversity Among Dominant Fungi Involved in Water Production from

Non-Traditional Water Resources

The dye bioremoval analysis by these ten isolates showed

high capacity to remove (300 mg l-1

) dye from aqueous

ecosystems. These results are in agreement with other authors

(White et al., 1997) who noted that heterotrophic fungi

belong to genus Mucor, Aspergillus, Penicillium were

capable of removing textile dyes. The mechanism of

removing dyes was stated to be due to reduction,

accumulation, mobilization and/or immobilization (White et

al., 1997).

3.2. Dye Removal in the Batch Liquid

Cultivation

Fig 1 showed that all isolates were active in bioremoval of

dye from simulated effluent aqueous solutions. The isolates

differed in their capacity to remove the dye. Isolates; 1, 7, 8,

9, 10 and 11 showed the capacity to remove from 72.9% to

85% of dye in 8 hours. The remained isolates either started

the bioremoval later than the previous strains by 8 hours

(isolate 3 and 4) or the speed of dye bioremoval (isolates 3

and 6) was much lower than other isolates. At the end of

incubation (28 hours) all isolates could remove between

12.51% and 85% of the dye. This showed the high capacity

of all tested isolates to remove the dye regardless the time

factor involved in this experiment.

The increasing of dye bioremoval by fungal strains was

mostly accompanied with the increase of fungal growth

(Fig1). The fungal biomass amounted 0.59, 0.78, 0.9, 0.96,

1.05, 1.10, 1.23, 1.26, 1.29, 1.29 and 1.34 g/ 300 ml for

isolates FRD1 to FRD8 on respectively. Strain FRD 8 gave

the maximum record of biomass dry weight (Fig2). The

increase in dry weight ranged between 2.5 to 6.5 times

comparing to the initial dry weight of the inoculums (200

mg). Similar results were obtained by Asher et al. (2008)

who stated that de-colorization of RB19 dye was carried out

by a variety of white-rot fungi. Singh (2008a) reported the

microbial degradation of Congo red by Gliocladium virens.

On the other side, dyes such as Congo red, Acid red, Basic

blue and Bromophenol blue, Direct green were successfully

bio-treated by fungus Trichoderma harzianum (Singh and

Singh, 2010). The biodegradation of Methylene blue,

Gentian violet, Crystal violet, Cotton blue, Sudan black,

Malachite green and Methyl red by few species of

Aspergillus was reported by Muthezhilan et al.(2008). The

biodegradation of three azo dyes (Congo red, Orange II and

Tropaeolin O) by the fungus Phaenerocheate

chrysosporium was reported by Cripps et al. (1990).

Revankar and Lele (2007) found that 73% of RB19 dye

(100 mgl-1

) could be removed in 8 hours by Ganoderma sp,

while in our study fungal isolates could remove (300 mgl-1

)

in 28h.

Changes in COD values by different isolates can be used

as a good evidence for progress of bioremediation process

(Ademorotti et al., 1992; Patheet al., 1995), consequently. In

this study the COD values measured at the end of experiment

(28h of inoculation) of the simulated effluent containing dye

and treated by ten fungal isolates are presented in Fig3. The

results showed that the fungal growth reduced the organic

load of the synthetic effluents by 56 to 81%. Fungal isolates

FRD 4, 8, 9 and 10 were the highest efficient strains in

removing (81% removal) of the organic dye contents of the

synthetic effluent. The rest of the isolates were capable of

removing up to 55% of the growth media dye contents. This

clearly showed that the fungal isolates included in this study

are good candidates for bioremediation of green textile dye.

These results are in agreement with previous studies stating

that fungi can play important role in detoxification of waste

water from hazardous organic pollutants (Muthezhilan et al.,

2008; Cripps et al. (1990) and Revankar and Lele (2007).

3.3. Activity of Lac Enzyme in

Biodegradation Process

Lac enzymes were reported to be found in eukaryotes, e.g.,

fungi, plants, and insects. Lac was used for the treatment of

different industrial effluents containing chlorolignin or

phenolic compounds (Bollag and Leonowicz, 1984; Brenna

and Bianchi, 1994; Jonssonet al., 1998; Ullahet al., 2000;

Bohmeret al., 1998; Call and IMucke, 1996). The lac

activities of the ten fungal strains used in this study are

presented in Fig 4. The fungal isolates FRD 1 and 6 had the

same pattern of lac enzyme activity. However, the

bioremoval of green dye by fungal biomass differed

significantly (Fig 4) being 34% with isolate FRD 6 compared

to (72%) removal by isolate FRD 1 after 28h incubation.

Four strains FRD2, FRD3, FRD4 and FRD6 were actively

produced lac enzyme throughout the incubation period.

Based on molecular identification which will presented later

in this study, two of these strains; FRD2 and FRD6 belonged

to Aspergillus terreus, whereas the other two strains; FRD3

and FRD4 were Aspergillus tubigenesis. These four strains

expressed steady increase in lac activity starting with early

incubation till the end of incubation 28h. The strains of A.

terreus FRD3 and FRD4 were distinguished, among all other

strains, with the highest lac activity. The results showed that

the lac activity of these selected strains is strain specific and

not species specific. Strain FRD1 that belonged to A. terreus

did not show lac activity as strain FRD2 and FRD3 which

belonged to the same species. Similar results were found with

strains A. tubigenesis FRD8 and FRD10. None of the strains

belonging to A. niger was distinguished as a strong lac

producer. The results of lac activity did not follow the same

trends obtained in dye bioremoval experiment presented in

(Fig 4). The strains which showed high dye removal capacity

namely, FRD8, FRD9, FRD10 and FRD11 were very poor in

lac production. This clearly showed that the dye bioremoval

takes place by more than one mechanism. It is likely that the

high bioremoval without high production of lac could be

mainly due to the high biosorption of the dye on fungal

biomass. The high biosorption of dyes by fungal growth was

reported before (Wafaa and El-ardy 2011, Wafaa et al., 2009,

Wafaa and Moawad 2010, Wafaa, 2006). The over production

of lac by other strains namely FRD2, FRD3, FRD4 and

FRD6 indicated that these strains contributed to remove of

AASCIT Journal of Biology 2015; 1(2): 15-24 19

the dye residues not only by biosorption but also by

biodegradation through lac production. It has been reported

that lac induce degradation of azo dyes and phonemic

compounds, which exist in commercial azo dyes (Nyanhongo

et al., 2002). Rodriguez et al. (1999) also reported that the

de-colorization of dye by Trametes hispida strain is mainly

ascribed to extracellular enzyme activity of lac that plays a

major role in de-colorization. The bioremediation of textile

dye effluents by fungi is a complex process that include

direct dye removal by fungal biomass (Heinflinget al. 1997;

Asher et al. 2008: Singh and Singh 2010: Tan et al., 2013)

and/or possible dye bioremediation by certain enzymes

existing in fungal cells such as Lac enzyme (Brenna and

Bianchi 1994; Nyanhongo et al., 2002).The reduction of lac

activity during the incubation time was associated with FRD

8, 10 and 11. However, these isolates had high efficiency in

dye bioremoval (85%). This may be attributed to the decrease

in dye concentration as a result of dye degradation which can

cause less induction of the enzyme due to decrease in

substrate. The presence of very low concentration of dye

within 28h could be the reason for decrease in the enzyme

activity. Similar results were reported by Kalme et al. (2007)

for LiP, lac and tyrosinase. On the other hand, Wafaa et al.

(2003b) reported that the rapid biosorption of dyes on the

intact fungal biomass caused a decrease of free dye

concentration in the medium.

3.4. Molecular Identification of Fungal

Isolates

Standard biochemical tests, and morphological

characteristics have conventionally been used for the

identification of fungal species, but these methods of

identification are costly, time-consuming, and require

special skills. In addition, these tests fail to identify near

fungal species due to lack of precise test for species

identification. An amplified fragment of 600 bp of

ITSrRNA was generated by PCR amplification (data not

shown).We used the RFLP technique using two restriction

enzymes (CfoI and MspI) to digest the targeted fragment of

ITS rRNA to differentiate among fungal isolates. The

digestion of the PCR product by enzyme CfoI could classify

the ten fungal isolates into three genetic profiles (Fig5).

Genetic profile I included (strains FRD 1, 2, 3 and 5);

genetic profile II contained (strains FRD 4, 7, 8, 10 and 11)

and genetic profile III (strain FRD 9). Strain FRD 6 had the

same genetic profile of group I (data not shown). The RFLP

of PCR product with enzyme Msp1 confirmed the results

obtained from digestion with CfoI (Data not show). The

obtained RFLP results confirmed the possibility of using

this part of ribosomal rRNA to differentiate among fungal

strains as reported previously by Colin et al. (1999) who

used the same part of ITS to identify Dermatophyte Fungi

strains. The use of RFLP analysis of ITS interagenic regions

of the rRNA repeat is a valuable technique both for

molecular strain differentiation of T. rubrum and for species

identification of common dermatophyte fungi. This study

showed that internal transcribed spacer region analysis

using polymerase chain reaction based on RFLP using CfoI

and MspI enzymes is useful for rapid differentiation among

the fungal isolates capable of removing textile dye.

Paolocci et al. (1997) successfully used the same technique

to identify commercially important black truffle fungi

(Tuber melanosporum). Results of phylogenetic tree (Fig 6)

confirmed the results of RFLP analysis. Strains FRD1 and 3

shared the genetic clade with reference strain of Aspergillus

terreus KALM 104, while strains FRD 7 and 11 similar to

Aspergillus tubigenesis and strain FRD 9 was similar to

Aspergillus niger. However there is no clear difference

between these last three strains in the phylogenetic tree,

although, strain FRD 9 had a unique RFLP pattern. This

may be due to mutation in into restrictions sites that made

strain FRD 9 differed than other strains or due to the high

similarity between species of Aspergillus niger and

Aspergillus tubigenesis. There are many papers on the

bioremediation of Azo-dyes but for our knowledge this is

the first report about the bioremediation of this specific dye

(green direct dye). Our results reflected the importance of

such fugal isolates to be use as a safe bio resource fungal

product to reduce the environmental pollution caused by the

industrial effluents. This study also identified three different

fungal species equipped with enzymatic machinery that can

play important role in textile dye bioremediation.

20 Wafaa M. Abd El-Rahim et al.: Biodiversity Among Dominant Fungi Involved in Water Production from

Non-Traditional Water Resources

Figure 1. Bioremoval of direct green dye by the biomass of fungal isolates throughout 28 hours of inoculation.

AASCIT Journal of Biology 2015; 1(2): 15-24 21

Figure 2. Fungi biomass accumulation after 28 hours in the medium amended with direct green dye.

Figure 3. Efficiency of DG dye bioremediation as indicated by COD reduction after 28 hours of inoculation.

22 Wafaa M. Abd El-Rahim et al.: Biodiversity Among Dominant Fungi Involved in Water Production from

Non-Traditional Water Resources

Figure 4. Activity of Laccase enzyme as expressed in OD associated with growth of fungal isolates.

AASCIT Journal of Biology 2015; 1(2): 15-24 23

Figure 5. RFLP of ITS rRNA fragments for fungal isolates using Cfo1

enzyme. From left to right, lanes: 1, 100 pb ladder, then FRD 1, FRD2,

FRD3, FRD4, FRD5, FRD7, FRD8, FRD9, FRD10, FRD11, and 100 bp

ladder.

Figure 6. Neighbor-Joining phylogenetic analysis of five representative

strains using 600 bp of ITS rRNA, Bootstrap probabilities of 1000 pseudo

replicates are indicated.

4. Conclusions

This study demonstrated that several fungal strains belong

to three different species of Aspergillus were highly active in

removing direct green textile dye from aquatic polluted

effluents through the two pathways of biosorption and

biodegradation. Strains FRD 1 and FRD7 were proven to be

fast bio-removing of the DG dye residue most likely by

biosorption mechanism. A. terrous strains FRD 3 and FRD4

were actively bio degrading dye due to their ability for lac

production. The consortium of these strains is recommended

as bioremediation agent for fast and save removal of the DG

dye residue.

Acknowledgement

Authors wish to thank Egypt/US joint funded for

sponsoring the project on “Developing of Enzymatic

Bioremediation Technology For Textile Dye Bioremoval”

which supported this research.

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