Process Biochemistry 39 (2004) 623–631

Treatment of tectilon yellow 2G by Chlorella vulgaris

E. Acuner, F.B. Dilek∗
Middle East Technical University, Environmental Engineering Department, Ankara 06531, Turkey

Received 12 August 2002; received in revised form 20 January 2003; accepted 10 April 2003

Abstract

Treatment of mono-azo dye, tectilon yellow 2G (TY2G), by Chlorella vulgaris was investigated. COD removal efficiencies were determined
as 69, 66 and 63% for the initial TY2G concentrations of 50, 200 and 400 mg/l, respectively, whereas acclimation of C. vulgaris caused them
to increase to 88, 87 and 88%, respectively. Absorbance spectral profiles obtained for unacclimated algae showed that the peak observed
initially at 450 nm disappeared and the one at 220 nm decreased remarkably while there was a new peak formation at 350 nm, indicating
the conversion of TY2G to an end product which was further confirmed as aniline by high-performance liquid chromatographic analysis.
In the case of acclimated algae, no aniline production was detected. The main mechanism of the removal was bioconversion in the case of
unacclimated algae and degradation in the case of acclimated algae. Moreover, the higher the initial algal concentration, the higher is the COD
removal efficiency achieved in a much shorter time.
© 2003 Elsevier Ltd. All rights reserved.

Keywords: Mono-azo dye; Chlorella vulgaris; Treatment; Toxicity; Acclimation

1. Introduction In recent years, the use of white-rot fungi in the decol-

Azo dyes account for the majority of synthetic dyes
used in the textile industry [1–4]. Release of azo dyes into
the environment from the effluents of the textile indus-
try has become a major concern in wastewater treatment
since some azo dyes may be mutagens or carcinogens [5].
Physical and/or chemical methods such as chemical coagu-
lation and/or ozonation [6–10], electrochemical destruction
[11,12] and sorption [13,14] have been used to achieve de-
colorization of the wastewater. However, implementation of
these methods may generate a significant amount of sludge
or may easily cause secondary pollution due to excessive
chemical usage or economic unfeasibility [15].
On the other hand, biological decolorization and degra-
dation of dyes remain more cost-effective [16]. However,
conventional biological treatment systems such as activated
sludge or lagoons fail to remove color from the textile indus-
try wastewaters due to the complex aromatic structures of
the dyes [17]. Anaerobic reduction and decolorization most
often generate aryl amines, which are generally more toxic
than the parent compounds [18,19].
∗ Corresponding author. Tel.: +90-312-210-5877;
fax: +90-312-210-1260.
E-mail address: fdilek@metu.edu.fr (F.B. Dilek).
orization of textile dyes has been investigated; however, the
requirement for the intrusion of other carbon sources is con-
sidered as the major drawback [20]. Therefore, there is still a
demand to develop more effective and more economical al-
ternative means of dye decolorization. In this respect, there
are some recent reports regarding degradation of azo dyes
by algae [15,21–24]. Certain algae (Chlorella vulgaris, C.
pyrenoidosa, Spirogyra and Oscillatoria tenuis) can degrade
a number of azo dyes to some extent, postulating that the
reduction appears to be related to the molecular structure of
the dyes and the species of algae used. Moreover, the dye
removal mechanisms suggested by these researchers show
some variations depending upon the dyes and species of al-
gae used. For example, Mohan et al. [15] proposed a mech-
anism of biosorption/respiration/photoconversion during the
treatment of reactive yellow 22 using Spirogyra species
whereas Jingi and Houtian [22] reported that Eriochrome
blueSE was biodegraded completely by C. vulgaris and O.
tenuis. On the other hand, Aziz and Ng [21] suggested a
mechanism of adsorption onto the surface of the algal cells
of mixed culture for the treatment of azo dye mixtures. Fur-
thermore, none of these studies has dealt with the effect of
acclimation of algae to dyes.
This study was aimed to investigate the treatment of a
mono-azo dye, namely tectilon yellow 2G (TY2G), with

0032-9592/$ – see front matter © 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0032-9592(03)00138-9
624 E. Acuner, F.B. Dilek / Process Biochemistry 39 (2004) 623–631
both unacclimated and acclimated cultures of C. vulgaris.
Dye toxicity reduction achieved during the dye treatment
was also sought.
2. Materials and methods
2.1. Dye used
Dye used in the experiments, namely TY2G (mono-azo
dye; Fig. 1) was obtained from the SAMUR Carpet Industry,
Turkey.
As can be seen from Fig. 1, TY2G has one N=N bond,
2-Cl (98% chloro compound), 2-SO3 , 1-NH2 and 1-OH
groups in its structure. According to the fact sheet of this
dye, provided by CIBA-GEIGY Corp., its toxicity level on
bacteria is 300 mg/l and its biodegradability is by 20–30%
as TOC.
2.2. Inocula
The alga, C. vulgaris, obtained from the Biology Depart-
ment of Hacettepe University, Turkey, was used in this study.
C. vulgaris was grown in several 1-l glass jars containing
growth medium in order to obtain stock algal culture to be
used during the experiments.
Activated sludge biomass used during the toxicity ex-
periments was taken from the aeration unit of the Ankara
Municipal Wastewater Treatment Plant.
3. Growth medium
The composition of the growth medium for C. vulgaris is
given in Table 1.
3.1. Experiments
3.1.1. Growth pattern of algae in medium containing dye
Growth of algae in a medium containing TY2G was fol-
lowed in 1-l glass jars. The reactors were placed in a trans-
parent waterbath at 25±1 ◦ C near the window and exposed
Fig. 1. Chemical structure of TY2G.
Table 1
Composition of growth medium [25]
Constituents Concentration (mg/l)
NH4 Cl 5241.7
KH2 PO4 4.9
K2 HPO4 5.0
MgSO4 ·7H2 O 5.0
CaCl2 ·2H2 O 3.357
Fe(NH4 )2 (SO4 )2 ·6H2 O 1.6
H3 BO3 0.50
(NH4 )6 Mo7 O24 ·4H2 O 0.40
MnSO4 ·H2 O 0.2805
ZnSO4 ·7H2 O 0.2097
CoCl2 ·6H2 O 0.13
CuSO4 ·5H2 O 0.010
Thiamine–HCl 2
Tris base 40.2
to the natural light and fluorescent light (18 W) mounted
about 30 cm above the jars. Air pumps and magnetic stirrers
provided gentle stirring. During the experiments, keeping
the fluorescent light source on for 12 h during the daytime
provided simulated field lighting conditions. C. vulgaris
was inoculated into jars containing 50, 200 and 400 mg/l
TY2G and growth medium ingredients (Table 1).The in-
oculum amount introduced to each jar was 500 ml of 400
mg/m3 stock algal culture. Samples were taken at reg-
ular intervals and filtered through 0.45 ␮m membranes
for chlorophyll-a analysis (Chl-a). A reactor containing
algae and growth medium without dye was provided as
control reactor. High-performance liquid chromatography
(HPLC) analyses for end product determination were also
performed.
3.1.2. Dye removal by unacclimated and acclimated algae
During the experiments with unacclimated algae, TY2G
was introduced to the reactors (1 l) containing growth
medium ingredients and 500 mg/m3 algae not previously ex-
posed to TY2G, to initial dye concentrations of 50, 200 and
400 mg/l. During the experiments with acclimated algae,
algae exposed to TY2G previously (i.e. obtained from the
reactors of the growth kinetic experiment of 19 days) were
used. The same operational conditions such as temperature,
lighting, aeration and stirring were provided as described
for the growth pattern experiments. Samples taken at regular
intervals were filtered through a 0.45-␮m membrane filter
and analysed for COD content and absorbance (A) values
at peak absorbance wavelengths (λmax ) of TY2G, namely
A450 nm and A220 nm . Spectral profiles from 190 to 700 nm
were also recorded before and after the algal treatments.
In addition, at the end of the experiments, dye adsorp-
tion on algal biomass was determined by extracting the dye
and measuring absorbance of the extract at 280 nm. One
cubic decimetre of samples containing algal cultures was
centrifuged and filtered through a 0.45-␮m membrane filter.
The algal filter cake was extracted with 20 ml of 5% NaOH
solution and washed twice with 10 ml of distilled water.
 E. Acuner, F.B. Dilek / Process Biochemistry 39 (2004) 623–631
Table 2
Composition of growth medium of activated sludge biomass [26]
Component Concentration (mg/l)
Proteous-peptone 407
NaCl 407.4
Na2 SO4 44.6
K2 HPO4 44.6
MgCl2 ·6H2 O 3.7
FeCl2 ·2H2 O 3.7
CaCl2 ·2H2 O 3.7
MnSO4 0.057
ZnSO4 0.046
CoSO4 0.049
CuSO4 0.076
The extract was adjusted to pH 7.5 with 0.1 N H2 SO4 and
diluted to 1 l with distilled water [27].
3.1.3. End product toxicity experiments
10 ml of activated sludge biomass suspension (1000 mg/l)
was inoculated into 500 ml Erlenmayer flasks containing in-
fluent and effluent samples of the dye removal experiments.
At the end of dye removal experiments with unacclimated
algae, the reactor contents were allowed to settle and the su-
pernatants were used as effluent samples. Growth medium
ingredients of activated sludge biomass (Table 2) were also
provided. Flasks were placed in a shaking incubator set at
a temperature of 25 ◦ C. Samples were taken every hour and
absorbance measurements were performed at 550 nm to fol-
low bacterial growth.
To eliminate dye-color contributions to the absorbance
measurements during experiments with influent samples,
absorbance values of dye solutions without biomass (i.e.
influent) were measured as well and these values were then
subtracted from the related sample measurements. In do-
ing so, no visual removal of dye by the activated sludge
throughout these toxicity experiments was taken into con-
sideration, and therefore correction was made only for the
initial absorbance values. This correction was not carried
out on effluent samples, as they bear no absorbance at
550 nm.
3.1.3.1. Analyses. COD measurements were carried out
according to an EPA-approved reactor digestion method
(for a COD range 0–1500 mg/l) using a HACH DR2000
Spectrophotometer (Model No.: 45600-02, Cole Parmer In-
strument Co., USA). For Chl-a analyses, a procedure given
for acetone extraction method (Method No.: 10200 H) as
described in standard methods (1998) was followed. Ab-
sorbance was measured with a Jenway 6105 UV/Vis spec-
trophotometer (Model No.: 6105, Jenway Ltd., England).
Spectral profiles were obtained using a Cary 105 Conc.
UV/Vis spectrophotometer (Model No.: ELCE093120,
Varian Co., Australia). End product formation was mon-
itored by HPLC (Shimadzu, LC-10AT vp) which was
equipped with a Nucleosil C18 column (4.6 mm×250 mm),
625
an LC-10AT vp solvent delivery module, an SCL-10Avp
system controller and an SPD-10Avp UV/Vis detector set
at 250 nm. The retention time of TY2G was 10 min. The
mobile phase was a 70:30 mixture of acetonitrile:water in
volume basis and applied at a flow rate of 0.75 ml min−1 .
Injection volume of sample was 20 ␮l.
In this study, all experiments and measurements were per-
formed in duplicate and the average of the duplicates was
used throughout the data analysis.
4. Results and discussion
4.1. Growth pattern of C. vulgaris in medium containing
TY2G
Growth of C. vulgaris in the presence of TY2G of differ-
ent concentrations, namely 50, 200 and 400 mg/l was fol-
lowed in terms of Chl-a content (Fig. 2). From the data be-
longing to the exponential phases of growth curves, specific
growth rates (µ) of C. vulgaris were calculated as 0.035,
0.030 and 0.021 per day for ascending dye concentrations.
These values fall below the value obtained for the control
experiment, i.e. 0.041 per day. Also, biomass levels reached
at the stationary phase of the growth showed a decreasing
trend with increasing dye concentration. On the other hand,
a lag phase duration (4 days) was observed to be the same
at all dye concentrations tested (Fig. 2). A similar lag phase
duration (4–6 days) for C. vulgaris in sewage culture was
observed by Lau et al. [28]. The same lag phase duration
for the control-algae case (Fig. 2) indicates no effect of dye
on the lag phase duration and, therefore, it can be said that
the lag phase duration is mainly governed by the autotrophic
growth process of the algae. Supportingly, Tarlan and Dilek
[29] reported that the algal lag phase is highly dependent
on the light intensity rather than the organic strength of
the wastewater as long as the nutrients required for the au-
totrophic growth are available.
COD removal obtained at the end of 14 days were quite
low with a value of 23, 13 and 7% for ascending dye con-
centrations. This COD removal, although low, was taken
as an indication of heterotrophic utilization of TY2G by
algae. Therefore, it can be stated that C. vulgaris grows
Fig. 2. Growth profile of C. vulgaris in the presence of TY2G.
626 E. Acuner, F.B. Dilek / Process Biochemistry 39 (2004) 623–631
Fig. 3. Results of HPLC analysis: (a) at the beginning of growth (400 mg TY2G/l itself); (b) at the end of growth (14 days); (c) at the time of 19 days;
(d) aniline (12.5 mg/l).
mixotrophically in the medium containing TY2G and it
would be worth to study the treatment of TY2G by C.
vulgaris.
HPLC analysis was also performed to monitor end prod-
uct formation. Fig. 3a and b present the results of analy-
ses obtained at the beginning and after C. vulgaris growth,
respectively. Fig. 3a shows that TY2G has two peaks; one
appears at about 2.5 min and the other at 4 min. A peak
observed at 2.077 min (Fig. 3b) refers to aniline produced,
which was confirmed by the injection of pure aniline solu-
tion to HPLC, resulting in a peak appearance at the same
detention time (Fig. 3d).
The degradation of aromatic amines formed after cleavage
of azo bonds by C. vulgaris was studied by Jingi and Houtian
[22]. They reported that C. vulgaris can utilize aniline, and
no other compounds were detected after treatment by C.
vulgaris. They claimed that C. vulgaris converted aniline
into some simple inorganic materials. Based on this literature
finding, it was thought that aniline produced in the present
experiments could be degraded by C. vulgaris if a longer
incubation period was allowed. Hence, the experiments were
continued for 5 more days to determine if aniline degradation
would occur. Fig. 3c illustrates the aniline peak observed
with an area of 52 190 on day 19th of incubation, which is
about 48.8% less than its previously detected value at the
end of day 14.
4.2. Dye removal by unacclimated and acclimated C.
vulgaris
Time course variations of COD (as normalized with re-
spect to initial COD) for different initial dye concentrations
are given in Fig. 4. COD removal by C. vulgaris occurs in
two phases: in the first phase which lasts about 10 and 6
h for unacclimated and acclimated algae, respectively, the
removal is rapid; and in the following second phase, the re-
moval slowed down reaching a stable value after about 30
and 150 h, respectively. Most of the COD was removed dur-
ing the first phase with unacclimated algae (Fig. 4a), whereas
it was removed in the second phase with acclimated algae
(Fig. 4b). Data obtained for the control reactor(s) proved
that there is no physical meaning of dye removal. COD re-
moval efficiencies achieved at the end of first and second
phases are presented in Fig. 5 for both unacclimated and
acclimated C. vulgaris. COD removal efficiencies obtained
with acclimated algae at the end of the experiment were sig-
nificantly higher than those obtained with unacclimated al-
gae. It is also observed that, for unacclimated algae, COD
removal efficiencies decreased with initial dye concentration
at the end of first phase (10 h); however, upon prolonged
incubation conditions (i.e. at the end of 200 h), the effect of
initial dye concentration on COD removal efficiency almost
disappeared. On the other hand, with acclimated algae, no
 E. Acuner, F.B. Dilek / Process Biochemistry 39 (2004) 623–631
Fig. 4. Time course variations of COD removal and Chl-a with (a) unacclimated and (b) acclimated C. vulgaris at different initial TY2G concentrations.
effect of initial dye concentration on COD removal was no-
ticed throughout the experimental period. When compared
with growth pattern experiment results, it was interesting to
observe that COD removals were high even in the case with
unacclimated algae. In fact, the only difference between the
growth pattern experiment and dye treatment experiment
Fig. 5. COD removal efficiencies with respect to initial TY2G concen-
trations.
627
with unacclimated algae was initial algal concentration, al-
though final algal biomass concentrations reached were al-
most same. Hence, the higher COD removal obtained with
unacclimated algae as compared to growth pattern experi-
ment can be taken as an effect of initial algal concentration
on COD or dye removal. In the case of the growth pattern
experiment, biomass concentration was initially 190–200
mg/m3 and then increased to about 230–260 mg/m3 within
10 days depending on the initial TY2G concentration. In the
case of unacclimated algae, the initial algal concentration
of 500 mg/m3 declined to 200 mg/m3 within the first 20 h
(Fig. 4a). This may indicate the lysis of the algal biomass,
because algae concentration was measured in terms of Chl-a.
When lysis occurs, a decrease in this parameter should be
expected since Chl-a would pass through the filter paper
while only algae which have not lysed would be captured
on the surface of the filter paper.
Interestingly, the total time required to reach the maxi-
mum COD removal or time beyond which no further re-
moval is observed (150 h) with acclimated algae is longer
than with unacclimated algae. Initial specific COD removal
628 E. Acuner, F.B. Dilek / Process Biochemistry 39 (2004) 623–631

Fig. 6. Time course variations of (a) A450 nm and A220 nm with unacclimated C. vulgaris and (b) A220 nm with acclimated C. vulgaris.

rates were determined for unacclimated algae as 148.1, 154.7

and 115.2 mg COD/mg algae/h, whereas for acclimated al-
gae, they were calculated as 41.6, 37.6 and 71 mg COD/mg
algae/h for ascending dye concentrations. Hence, it can be
concluded that the initial COD removal rates observed were
lower with acclimated algae than with unacclimated algae
at all dye concentrations.
Variation of absorbance values at peak absorbance wave-
lengths, A450 nm and A220 nm , with time is also followed
for unacclimated and acclimated algae as shown in Fig. 6.
A450 nm almost disappeared for all concentrations within the
first 15 h with unacclimated algae (Fig. 6a). A220 nm re-
moval was most efficient during the first 6 h, however, un-
like A450 nm , almost complete removal could not be achieved
even at the end of the experiment (Fig. 6a). Hence, only
A220 nm measurements were conducted for acclimated algae
(Fig. 6b). A220 nm removal efficiencies were found to be 85,
66 and 63% with unacclimated algae and 90, 85 and 83%
with acclimated algae for the initial dye concentrations of
50, 200 and 400 mg TY2G/l, respectively.
Spectral analyses of TY2G before and after algal treat-
ment were also performed (Fig. 7a) in order to understand Fig. 7. Spectral analysis obtained (a) before and after treatment of 400
if any intermediate or end product formation has occurred mg TY2G/l (b) for pure aniline.
 E. Acuner, F.B. Dilek / Process Biochemistry 39 (2004) 623–631 629

Fig. 8. HPLC results obtained after treatment with (a) unacclimated and (b) acclimated C. vulgaris.

during treatment by unacclimated C. vulgaris. The peak ob-
served at 220 nm decreased quantitatively and the peak ob-
served at 450 nm almost disappeared. However, new peak
formation took place at 350 nm. This new peak probably be-
longed to the aromatic amine, aniline, which is the expected
end product of the azo dye degradation. The spectral anal-
ysis of pure aniline solution presented in Fig. 7b confirmed
that the end product was aniline as it also yielded a peak at
350 nm.
In order to understand how much dye the algal cells had
adsorbed, an alkaline extraction experiment was performed.
Unacclimated algal cells for ascending dye concentrations
adsorbed 11, 9 and 8.75% of dye, whereas acclimated ones
adsorbed 10, 9.25 and 9% of dye. This indicated that adsorp-
tion was lower than about 10% and it should not be taken
into consideration as a main mechanism of TY2G removal
by C. vulgaris. Similarly, Mohan et al. [15] observed that
10% of azo dye removal by algae was through the biosorp-
tion in the case of removal of reactive yellow 22 by Spir-
ogyra species. In addition, however, they claimed that once
the dye molecules adsorbed onto the surface of the algal
cells, they participated in respiration/photoconversion reac-
tions (bioconversion). Dye remaining in the aqueous phase
then tended to adsorb onto the surface of polymers, which
were released as metabolic intermediates with excellent co-
agulation capacity, and then settle (biocoagulation) [15]. In
contrast, Aziz and Ng [21] reported that adsorption onto
the surface of the algal cells was the primary mechanism of
color removal from textile wastewater.
HPLC analyses were run to confirm the end product in
order to be able to suggest a removal mechanism. A com-
parison of Fig. 3a and Fig. 8a revealed that unacclimated
algae converted all TY2G to aniline. As can be seen from
Fig. 8a, at the end of the experiment with unacclimated al-
gae, an aniline peak was observed at 2.10 min. Hence, the
main mechanism of TY2G removal by C. vulgaris was con-
sidered to be bioconversion. As supportive evidence, Jingi
and Houtian [22] stated that in the reduction of azo dyes by
algae, the azo bridge is degraded by azo reductase and aro-
matic amine arise as a cleavage product. They proposed a
degradative pathway shown in Fig. 9. This degradative path-
way is mostly similar to the reduction mechanism of bacteria

Fig. 9. Dye removal mechanism of algae [22].

630 E. Acuner, F.B. Dilek / Process Biochemistry 39 (2004) 623–631

Table 3
Aniline production by unacclimated C. vulgaris at the end of 100 h
TY2G concentration (mg/l) Peak area Aniline concentration (mg/l) Theoretical COD of aniline (mg/l) COD remaining (mg/l)

50 5700 35 60 62
200 20843 128 225 255
400 39800 244 546 555

[30,31]. In this study, no other intermediate product, which ment. For 50, 200 and 400 mg TY2G/l dye concentrations,
could be detected with HPLC column used, was observed. the values of µ were estimated as 0.131, 0.065 and 0.053
The only end product formed was aniline as shown through h−1 , respectively, whereas the value for without dye (con-
the comparison of COD values remaining at the end of the trol) was 0.3 h−1 . All dye concentrations applied to acti-
experiment with the aniline concentrations and their corre- vated sludge biomass caused a decrease in the final amount
sponding theoretical COD values. The remaining COD of of biomass reached. In addition, as dye concentration in-
about 555 mg/l was derived from aniline produced (Table 3). creased, the value of µ decreased. This indicated that with
This can be confirmed by the theoretical COD (526 mg/l) the application of 50 mg TY2G/l and higher concentrations,
of 244 mg/l aniline. Therefore, all the remaining COD was inhibition of the growth of activated sludge biomass was
due to aniline. There may be other intermediate compounds clear as compared to the control in which no addition of
containing a single bond (–N–N–) before aniline formation TY2G was performed.
as shown in Fig. 8, but it could not be detected. No interme- The growth curves of the activated sludge biomass, ob-
diate other than aniline was observed even at the end of day tained in a medium containing the effluents of dye treatment
2 (data not shown). On the other hand, no aniline formation with unacclimated algae are presented in Fig. 10b. The cal-
was detected at the end of the experiment with acclimated culated µ values are 0.210, 0.480 and 0.560 h−1 for the
algae (Fig. 8b). Aniline was probably first produced and then
degraded completely by C. vulgaris within the experimental
time period (i.e. 200 h). This complete degradation of azo
dyes by algae was also suggested by Jingi and Houtian [22]
(Fig. 9), but they did not suggest any time period required
for this complete degradation. Although they stated that the
activity of azo reductase enzyme was increased remarkably
on acclimation of algae to dye, they did not link this ability
of algae (i.e. degrade the dye completely) to its acclimation
state. Therefore, it would not be wrong to claim that C. vul-
garis can degrade aniline effectively and turn it into some
simple inorganic material in a shorter time of period if accli-
mated. This is also supported by the results of growth pat-
tern experiment in which only 50% aniline degradation had
been observed in about 5 days if algae were not acclimated.

4.3. Toxicity experiments

Considering the end product, aniline, formation during

treatment with unacclimated algae and hence the possibility
of toxicity creation, effluent samples were subjected to tox-
icity analysis using activated sludge microorganisms. The
reason for selecting the activated sludge microorganisms to
test the toxicity was twofold. Activated sludge is a mixed
culture containing a variety of microorganisms and hence
represents a wide range of biota. In addition, in the case of
the treatment of TY2G containing wastewater with unaccli-
mated algae, further biological treatment to remove aniline
may be required and therefore the biodegradability or toxic-
ity of aniline-containing effluents in activated sludge plants
should be known.
Fig. 10a presents the growth curves of the activated sludge Fig. 10. Growth curves of activated sludge biomass at different TY2G
biomass at different dye concentrations before algal treat- concentrations (a) before and (b) after algal treatment.
 E. Acuner, F.B. Dilek / Process Biochemistry 39 (2004) 623–631
ascending initial dye concentrations. The biomass amount
reached at the end of the exponential growth phase in the
case of effluent of 50 mg/l TY2G-containing wastewater was
the same as in the control reactor and higher than the control
value in cases of 200 and 400 mg/l TY2G effluents. It can
then be stated that the effluents of the algal dye treatment are
not toxic in spite of the end product existence. Increase in µ
values with increase in end product aniline concentration to-
gether with increase in final biomass concentration reached
may suggest the ready utilization of aniline as substrate by
the activated sludge microorganisms.
5. Conclusion
• C. vulgaris was effective in the treatment of TY2G, es-
pecially when acclimatized. The effect of initial TY2G
concentration on the removal was not significant, within
the studied range.
• The mechanism of TY2G removal was not adsorption
onto the algal biomass, but bioconversion or degradation.
TY2G was found to be first converted to aniline, which is
further degraded by C. vulgaris upon prolonged exposure
times in the case of unacclimated algae while no aniline
was detected in case of acclimated algae.
• The toxicity of the effluent of TY2G treatment using un-
acclimated algae was lower than that of untreated TY2G.
End product aniline detected at the end of dye treatment
served as a substrate for the activated sludge microorgan-
isms, with a stimulating effect on their growth rate.
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