Bulletin of Environmental Contamination and Toxicology (2019) 102:795–801

https://doi.org/10.1007/s00128-019-02599-w

Toxic Effects of Selected Textile Dyes on Elemental Composition,

Photosynthetic Pigments, Protein Content and Growth of a Freshwater
Chlorophycean Alga Chlorella vulgaris
Samchetshabam Gita1 · S. P. Shukla1   · Neelam Saharan1 · Chandra Prakash2 · Geetanjali Deshmukhe3

Received: 22 October 2018 / Accepted: 22 March 2019 / Published online: 29 March 2019
© Springer Science+Business Media, LLC, part of Springer Nature 2019

Abstract
Toxicity of three textile dyes—Optilan yellow, Drimarene blue and Lanasyn brown, was evaluated in a green alga Chlorella
vulgaris. The unialgal populations of the alga showed a concentration-dependent decrease in specific growth rate and pig-
ments after exposure to graded concentrations of above dyes. The elemental profile (C, H, N, S) of the treated and untreated
cells showed a change which was evident from a significant decrease in the quantity of elements after exposure to the dyes.
The observations provide convincing evidence that the textile dyes inhibited the growth, pigment and elemental composition
of the algal cells. The findings of the present investigation will contribute to gaining a better understanding of the impacts
of textile dyes on ecologically important aquatic organisms.

Keywords  Textile dye · Chlorella vulgaris · Growth · Protein · Pigments

In India, the dyestuff sector is one of the core chemical
industries as well as the second highest export segment in
the chemical industry. The Indian dyestuff industry is made
up of about 1000 small-scale units and 50 large organized
units which produces around 130,000 tonnes of dyestuff
(Naik et al. 2013). Maharashtra and Gujarat account for 90%
of dyestuff production due to the availability of raw materi-
als and the dominance of the textile industry in the region
(Naik et al. 2013). The colorization process that requires a
high amount of water generates a considerable volume of
dye wastewater. In the textile industry, during the dyeing
and finishing operations, up to 200,000 tons of textile dyes
are lost to effluents in the environment every year due to the
inefficiency of the dyeing process because all the dyes are
not adsorbed to the surface of the dying materials (Chequer
* S. P. Shukla
spshukla@cife.edu.in
1 tile wastewater ultimately lead to pronounced damage to the
Aquatic Environment and Health Management Division,
ICAR-Central Institute of Fisheries Education, Mumbai,
Maharashtra 400061, India
2
Aquaculture Division, ICAR-Central Institute of Fisheries
Education, Mumbai, Maharashtra 400061, India
3
FRPHM division, ICAR-Central Institute of Fisheries
Education, Mumbai, Maharashtra 400061, India
et al. 2013). In another study, it was estimated that glob-
ally 280,000 tons of textile dyes are discharged in the dye-
containing industrial effluents every year (Jin et al. 2007).
The wastewater which is released from the textile plants or
industries after dying is classified as the most polluting agent
of all the industrial sectors, due to the volume generated
and discharged as well as the effluent toxicity (Shukla et al.
1992; Mansour et al. 2012). The discharge of synthetic dyes
into the aquatic system has generated much concern due to
reported genotoxic, mutagenic, teratogenic and carcinogenic
effects (Chowdhury and Saha 2010). In general, dyes have
low toxicity towards mammals and aquatic organisms, but
products formed by their biodegradation, mainly aromatic
amines from the anaerobic reduction of dyes, can be harmful
(Pinheiro et al. 2004). Some dyes are not only highly toxic,
carcinogenic and mutagenic but they also lead to a decrease
in light penetration and therefore, photosynthetic activity
causing oxygen deficiency (Przystaś et al. 2012). Also, the
synergistic effects caused by other pollutants present in tex-
aquatic environment (Samchetshabam et al. 2017).
Little research has been done to study the toxicity of
textile dyes on microalgae as compared to other pollutants
such as heavy metals. However, given the rapidly growing
dye industries, an assessment on the toxicity of textile dyes
towards aquatic algae is important to design the future course

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796
of action in regards to the environment safety issues. The
awareness in the dye manufacturing sector has addressed
various issues pertaining to the environment and the prod-
ucts with least impacts on the environment are developed
and marketed. However, evaluation of the effects of textile
dyes on aquatic organisms will stimulate interest among the
users for the adoption of a proper treatment technology.
Materials and Methods
The algal sample was obtained from Micro-Algal laboratory,
Aquatic Environmental Management, ICAR-CIFE, Mum-
bai, India and study was conducted in the same place. The
pure culture of C. vulgaris was sub-cultured in sterile BG-11
media (Allen 1968) under photoautotrophic conditions, and
was maintained in the plant growth chamber with the illu-
mination of 3500 ± 100 lx using compact fluorescent lamps
(Philips, 23 W). The photoperiod was fixed at 16:8 h light
and dark periods and temperature maintained at 24 ± 2°C.
The cultures were shaken twice a day to ensure the proper
mixing of the algal suspension.
Three different textile dyes namely Optilan yellow (acid
dye), Lanasyn brown (acid dye) and Drimarene blue (reac-
tive dye) were gifted by Achroma Ltd. (Mumbai, India).
Range finding experiment was conducted (OECD 1984) to
determine the concentration range of the dye. The concen-
tration range of 10, 20, 30, 40, 50 ppm was prepared from
the result of the range finding experiments. All the tests
were conducted in triplicate for each dye concentration. To
determine the range of environmental sample concentration,
samples were collected from textile processing units. The
dye-house approached for the sample collection uses a pro-
cedure for the dying of the cloths without any pre-treatment
such as bleaching and mercerizing through chemicals. The
samples were collected at the end of the dying process in
pre-washed amber color bottles and brought to the labora-
tory for further analysis. Optical density of collected sample
was measured using Double beam UV–Visible spectropho-
tometer (MOTRAS Scientific, New Delhi) and concentration
was determined from standard graphs of same dye.
96 h toxicity experiment was conducted according to
OECD guidelines 201 (OECD 1984), with certain modifi-
cations like the duration of exposure for 96 h. The exponen-
tially growing algal culture was harvested by centrifugation
and re-suspended in dyes solutions of graded concentrations
(10, 20, 30, 40, 50 ppm) in the medium. The culture density
for all the experiments was maintained at 3 × 105 cells ­mL−1.
Three replicates at each test concentration including control
were incubated in the plant growth chamber for 96 h under
the photoautotrophic conditions as mentioned above. The
cultures were manually shaken twice a day to re-suspend
any settled cells. Samples were analyzed every 24 h time
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Bulletin of Environmental Contamination and Toxicology (2019) 102:795–801
interval. A volume of 15 mL algal cells was centrifuged
(Eltek Microprocessor High-speed Research refrigerated
centrifuge, MP 400 R, India) at 5000 rpm for 10 min. The
supernatant containing the dye was discarded, and algal
pellets were washed thrice with 15 mL sterilized double-
distilled water to remove the dye molecules adsorbed on the
surface of cells. The direct optical density was measured at
650 nm wavelength using a software (Scanalyse) operated
Double beam UV–Visible spectrophotometer (Tungsten and
halogen lamps; Dual Silicon photodiode detector; blazed
holographic grating in Czerny-Turner mounting for mono-
chromator with a range of 190–1100 nm. The instrument
was calibrated before the measurements using potassium
dichromate solution prepared by dissolving 57.0 mg of the
compound in 0.005 M, ­H2SO4, followed by maintaining of
the volume to 1000 mL. The values of Absorbance (A) (1%
1 cm) for the wavelengths 235, 257, 313 and 350 nm were
measured to ensure that the values are within the acceptable
range.
Specific growth rate (K) and Generation time (x) were
calculated using the formula given by Kratz and Myers
(1955):
) 2.303 log Nt − log N0
K day−1 =
(
Tt − T0
where, ­N0 is the initial optical density at 650 nm for C. vul-
garis at ­T0 and ­Nt is the final number optical density at time
­Tt when culture is in exponential phase.
The generation time or mean generation time (days) was
calculated using the formula:
In (2) 0.693
x= =
k k
For the estimation of pigments, a volume of 15 mL algal
suspension was collected in a centrifuged tube after wash-
ing with 15 mL sterilized double-distilled water to remove
the dye molecules adsorbed on the surface of cells and cen-
trifuged at 5000 rpm for 10 min (Eltek, India). 15 mL of
N, N-dimethylformamide (DMF) was added to the remain-
ing pellets, and the suspension was incubated under dark
conditions for 24 h at the room temperature (24 ± 2°C).
After incubation, the solution was centrifuged for 10 min at
5000 rpm, and supernatants were collected in clean tubes,
and optical densities were measured at the prescribed wave-
length of 664 nm and 461 nm for estimation of total chloro-
phyll and carotenoid respectively. The pigments chlorophyll
(Moran 1982) and carotenoids (Chamovitz et al. 1993) were
estimated using the following formula:
Total chlorophyll = (μg∕mL) = O.D.664 × 11.92
[ ( )]
Carotenoid (μg∕mL) = O.D.461 − 0.046 × O.D.664 × 4
Bulletin of Environmental Contamination and Toxicology (2019) 102:795–801 797

Protein content was estimated by taking one mL of
sample at every 24 h intervals during the test period using
bovine serum albumin (BSA) as a standard protein follow-
ing the method of Lowry et al. (1951). Data for 96 h median
effective concentration (­ EC50) of three different textile dyes
for C. vulgaris was calculated from cell number using probit
analysis (Finney 1971), SPSS 22.0.
Effect of dye on carbon, nitrogen, hydrogen, and sulfur
content of C. vulgaris was analyzed using the CHNS Ana-
lyzer (Elementar, VarioMICRO, India). Algal sample after
96 h of dye exposure [both in controls and treatments (only
­EC50 value)] were washed with distilled water and centri-
fuged. Then, algal biomass was dried in the oven at 60°C
for overnight. The analytical conditions of CHNS analyzer
were: Carrier gas: Helium (Flow rate 200 ± 10 mL min−1),
Combustion gas (Oxygen) flow rate : 10 ± 2  mL  min−1
(When no combustion), 30 ± 2 mL min−1 (During the onset
of combustion), Detector: Thermal Conductivity Detector,
Combustion tube temperature: 1150°C, Reduction tube
temperature: 850°C, Thermal conductivity detector temp:
59–60°C, Pressure: 1200 ± 50 mbar.
The result obtained after the toxicity experiments were
statistically analyzed using SPSS 22.0. One-way analysis of
variance (ANOVA) was done to determine the significant
deferences among the mean and when differences observed
the means were compared by Duncan multiple range tests,
at a level of significance of 0.05 (p < 0.05). Results are pre-
sented as a mean ± standard error. Independent t-test was
done to analyze percentage elemental composition of the C.
vulgaris after exposure to dye.
Results and Discussion
The concentration of the dyes discharged to the environ-
ment ranged from 20 to 200 ppm. The concentration of
the dyes used varied in a wide range due to the varying
dyeing procedures, depending mostly on the type of fabric
and the number of cycles involved in the complete process.
Dye concentration range used in the present investigation
was selected on the basis on range finding test where the
organism was exposed to graded concentration of the dyes
10–200 ppm and, ­EC50 values were calculated for individual
dyes using the cell number of C. vulgaris. The E ­ C50 ranged
from 34.13 ppm (Drimarene blue), 36.27 ppm (Lanasyn
brown), to 46.40 ppm (Optilan yellow). Considering the
­EC50 values, we selected 10–50 ppm concentration range
for toxicity evaluation. The method and procedure used for
quantification of the dyes showed an excellent sensitivity
up to 0.01 ppm. In spite of the fact that the techniques such
as GC–MS and HPLC provide higher sensitivity, the spec-
trophotometric method is widely used for quantification of
coloured compounds including dyes (Moosvi et al. 2005;
Yaseen and Scholz 2018a, b).
After exposure period of 96 h, the effects of the various
concentrations of three textile dyes on the specific growth
rate, ­EC50, generation time, protein content, pigments con-
tent and elemental composition for C. vulgaris was studied.
It was observed that with an increase in the concentration
of dye, the specific growth rate (SGR) of C. vulgaris signifi-
cantly decreased for all three dyes up to certain concentra-
tion after that no growth was detected, i.e. 100% growth
suppression was noticed (Fig. 1a). The concentration higher
than 30 ppm completely suppressed the growth in all the
three dyes tested. A significant (p < 0.05) decrease in the
specific growth rate and a corresponding increase in the
generation time for all the concentrations of the dyes was
recorded (Figs. 1, 2, 3). Based on the SGR, the percent-
age inhibition of growth was calculated and a significant
difference (p < 0.05) in percentage growth inhibition was
observed among the various concentrations of all the three
dyes exposed groups. The highest percentage inhibition of
SGR was 67% for Drimarene blue, 68% for Optilan yel-
low and 66% for Lanasyn brown occurred at 30 ppm for
all three dyes, and after that, growth was totally inhibited
for all three dyes (Figs. 1, 2, 3). Hu and Wu (2001) also

a b c
0.3 10
Generaon me (days)

w d 150
Inhibion of SGR (%)

x D D
SGR (Day-1)

0.2
y c 100
5 b C
z B
0.1 a 50 A
0 0 0

Dye concentraon (ppm) Dye concentraon (ppm) Dye concentraon (ppm)

Fig. 1  Effect of various concentrations of Drimarene blue on specific with different superscripts in the alphabetical symbol are significantly
growth rate (a), generation time (b) and % inhibition of SGR (c) of (p ≤ 0.05) different from each other
C. vulgaris. Data are represented in mean ± SE, n = 3. The data labels

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798 Bulletin of Environmental Contamination and Toxicology (2019) 102:795–801

a b c
0.3 120
x 10
c d D D

Inhibi on of SGR (%)

Genera on Time (Day)
0.25 100
y 8
80 B C
SGR (Day-1)

0.2 b
6
0.15 z z a
60
A
0.1 4 40
0.05 2 20
0 0
0

Dye Concentra on (ppm) Dye concentra on (ppm) Dye concentra on (ppm)

Fig. 2  Effect of various concentrations of Optilan yellow on specific with different superscripts in alphabetical symbols are significantly
growth rate (a), generation time (b) and % inhibition of SGR (c) of (p ≤ 0.05) different from each other
C. vulgaris. Data are represented in mean ± SE, n = 3. The data labels

a b c
10
Genera on tome (day)

0.3 w d 120

Inhibi on of SGR (%)

0.25 8 100
D D
x C
SGR (day-1)

0.2
y 6 c 80
0.15 b 60 B
z 4 a A
0.1 40
2 20
0.05
0 0 0

Dye concentra on (ppm) Dye concentra on (ppm) Dye concentra on (ppm)

Fig. 3  Effect of various concentrations of Lanasyn brown on specific with different superscripts in the alphabetical symbol are significantly
growth rate (a), generation time (b) and % inhibition of SGR (c) of (p ≤ 0.05) different from each other
C. vulgaris. Data are represented in mean ± SE, n = 3. The data labels

reported the effect of azo dye on the growth of cyanobac-
terium, Anabaena sp. and results indicated that the growth
of Anabaena sp. was inhibited in the medium containing
dye. The degree of inhibition increased from 50% to 81%
with an increase in the concentration of dye (0–50 mg/L).
Textile dye wastewater consists of several components such
as heavy metal, suspended solid etc. All these compounds
including the dye itself may be the cause for the reduction
in growth and biomass productivity of the microalga with
increasing concentrations of the dye.
EC 50 (96  h exposure) of three dyes for C. vulgaris
was found to be 34.13 ± 1.23  ppm (Drimarene blue),
46.40 ± 2.09  ppm (Optilan yellow), 36.27 ± 1.48  ppm
(Lanasyn brown), respectively at 95% confidence limit.
The protein level in all the three different dyes concentra-
tion treated algal groups showed a significant reduction.
Further significant increase in percentage protein inhibition
was noticed with increased in dye concentrations (Fig. 4).
Dwivedi (2013) reported that further increase in the concen-
tration of Congo red dye in the growth medium resulted in
suppression of protein content. It was revealed that protein
content at the highest concentration of dye (100 mg/L) is
lowest (33.87 µg/mL). Hu and Wu (2001) also reported that
an increase in the concentration of azo dye ­RP2B inhibits
the protein synthesis of the cyanobacterium Anabaena sp.
The pigments, chlorophyll, showed a significant reduc-
tion (p < 0.05) in pigment content in all the three dyes with
increased in concentration of dyes (Fig. 5). Mathias and
Rilwan (2013) revealed that Chlorophyll-a content, cell
density and dry weight of selected algae was negatively
affected by the dye indigo blue present in the effluent. Pres-
ence of high concentration of dye in water might affect the
light transmitted in the water, ultimately affecting the photo-
synthesis and growth of aquatic cyanobacteria (Hu and Wu
2001). Similar studies also reported by Dwivedi (2013) and
Mahalakshmi et al. (2015). Based on the above literature
cited, all the findings agreed with the present study.
On applying t-test, it was noticed that for Optilan yel-
low the percentage value of carbon, hydrogen, and sulfur
content differed significantly (p < 0.05) whereas no signifi-
cant (p > 0.05) difference was noticed in nitrogen content. In
case of Drimarene blue and Lanasyn brown, the percentage
value of hydrogen and sulfur content differed significantly
(p < 0.05) and no significant (p > 0.05) difference was found
in nitrogen and carbon (Table 1). There is very few reports
available on effects of dyes on elemental composition of

13
Bulletin of Environmental Contamination and Toxicology (2019) 102:795–801 799

0.14 30 D
0.12 a D
25 BC

Protein inhibition (%)

bc cd
Protein (mg/ml) 0.1 cd de de BC
20
0.08 A
0.06 15

0.04 10
0.02 5
0
0
10 20 30 40 50
Dremarene Blue dye concentration Dremarene Blue dye concentration
(ppm) (ppm)

0.12 a ab 30 E
bc cd

Protein inhibition (%)

0.1 de 25 D
Protein (mg/ml)

de
0.08 20 C
0.06 15 B
0.04 10 A
0.02 5
0 0
10 20 30 40 50
Optilan Yellow dye concentration Optilan Yellow dye concentration
(ppm) (ppm)

0.14 a C CD
40
0.12 b b 35
Protein inhibition (%)
Protein (mg/ml)

0.1 c d
d 30 B
0.08 25
0.06 20
A A
0.04 15
10
0.02
5
0
0
10 20 30 40 50
Lanasyn Brown dye concentration Lanasyn Brown dye concentration
(ppm) (ppm)

Fig. 4  Effect of various concentrations of dyes on the protein content (mg/mL) and protein inhibition (%) of C. vulgaris. Data are represented in
mean ± SE, n = 3. The data labels with different superscripts are significantly (p ≤ 0.05) different from each other

algae. However, effects of pesticides (paraquat and atrazine)
on Chlamydomonas moewusii and C. reinhardtii decrease
the carbon–nitrogen ratio significantly in treatments group
compared to control (Fernandez-Naveira et al. 2016).
Among the three dyes, Drimarene blue was found to be
most toxic, and others were medium to moderately toxic
to the microalgae species. However, in view of the toxic
compounds formed after degradation in the environment
such as aromatic amine, benzidine, and its derivatives, a
complete study involving the assessment of toxicity of the
products formed through degradation of these dyes will give
a better insight into the mechanism of toxicity. In this study,
the ubiquitous alga C. vulgaris was used as a model sys-
tem for assessing the toxicity on algae (primary producers)

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800 Bulletin of Environmental Contamination and Toxicology (2019) 102:795–801

Chlorophyll Carotenoid Chlorophyll Carotenoid

70

Pigment content (µg mL-1)

10 a C

% inhibition of pigment
b 60 e
8 dC
c 50 cC
6 cd d 40

content
d b
4 30 B
A B C D DE E 20 a
2 A
10
0 0
10 20 30 40 50
Dremarene Blue dye concentration Dremarene Blue dye concentration
(ppm) (ppm)

Chlorophyll Carotenoid Chlorophyll Carotenoid

8 60
Pigment content (µg mL-1)

7
a eE
50 cd

% inhibition of pigment
6 c C CD
b bc 40
5 cd a b
de B
e

content
4 30
A
3 A B B C D 20
2 C
1 10
0 0
10 20 30 40 50
Optilan Yellow dye conc. (ppm) Optilan Yellow dye conc. (ppm)

Chlorophyll Carotenoid Chlorophyll Carotenoid

8 60
Pigment content (µg mL-1)

a D
% inhibition of pigment

7 b 50 D d
b
6 c
d 40 c
5 C
content

4 e 30
AB b
3 A 20 A
B B C a a
2 D D 10
1
0 0
10 20 30 40 50

Lanasyn Brown dye conc. (ppm) Lanasyn Brown dye conc. (ppm)

Fig. 5  Effect of various concentrations of three different dyes on the pigment content and % inhibition of pigment of C. vulgaris. Data are repre-
sented in mean ± SE, n = 3. The data labels with different superscripts are significantly (p ≤ 0.05) different from each other

Table 1  Effect of textile dye on Elements Percentage ­content#

major elements content of C.
vulgaris (data are represented in Controls Drimarene blue Optilan yellow Lanasyn brown
mean ± SE, n = 3)
Nitrogen 8.603A ± 0.494 7.938A ± 0.106 7.690A ± 0.051 8.251A ± 0.111
Carbon 43.826A ± 0.182 43.268A ± 0.245 42.534B ± 0.306 43.752A ± 0.191
Hydrogen 4.987A ± 0.145 3.884B ± 0.140 4.046B ± 0.041 4.047B ± 0.047
Sulfur 1.223A ± 0.154 0.000B ± 0.000 0.000B ± 0.000 0.05B ± 0.039
#
 mean values with different superscripts are significantly (p ≤ 0.05) different from each other

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Bulletin of Environmental Contamination and Toxicology (2019) 102:795–801 801
however; further studies on toxicity using a larger number
of pure cultures of algae are recommended. This investiga-
tion provides useful baseline data for a better understand-
ing of the mode of toxicity of textile dyes on algae. Given
a scarcity of data on toxic effects of textile dyes and dye
house wastewater on algae, this report will facilitate further
investigations on effects of textile dyes on aquatic organisms
in general and microalgae in particular.
Acknowledgements  The authors are thankful to the Director, ICAR-
Central Institute of Fisheries Education for providing the necessary
facilities. S. Gita acknowledges the valuable inputs from the advisory
committee members. Archroma, Mumbai is acknowledged for gifting
the dyes. This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
References
Allen MM (1968) Simple conditions for growth of unicellular blue-
green algae on plates. J Phycol 4:1–4
Chamovitz D, Sandmann G, Hirschberg J (1993) Molecular and bio-
chemical characterization of herbicide-resistant mutants of cyano-
bacteria reveals that phytoene desaturation is a rate-limiting step
in carotenoid biosynthesis. J Biol Chem 268:17348–17353
Chequer FMD, de Oliveira GAR, Ferraz ERA, Cardoso JC, Zanoni
MVB, de Oliveira DP (2013) Textile dyes: dyeing process and
environmental impact. In Eco-friendly textile dyeing and finish-
ing. InTech. https​://doi.org/10.5772/53659​
Chowdhury S, Saha P (2010) Sea shell powder as a new adsorbent
to remove Basic Green 4 (Malachite Green) from aqueous solu-
tions: Equilibrium, kinetic and thermodynamic studies. Chem Eng
J 164:168–177
Dwivedi S (2013) Effect of textile dyes on Spirulina platensis. J Chem
Pharm Res 5:66–80
Fernández-Naveira A, Rioboo C, Cid A, Herrero C (2016) Atrazine
induced changes in elemental and biochemical composition and
nitrate reductase activity in Chlamydomonas reinhardtii. Eur J
Phycol 51:338–345
Finney DJ (1971) Probit analysis, 3rd edn. Cambridge University Press,
Cambridge
Hu TL, Wu SC (2001) Assessment of the effect of azo dye RP2B on
the growth of a nitrogen fixing cyanobacterium–Anabaena sp.
Bioresour Technol 77:93–95
Jin X, Liu G, Xu Z, Yao W (2007) Decolorization of a dye industry
effluent by Aspergillus fumigatus XC6. Appl Microbiol Biotechnol
74:239–243
Kratz WA, Myers J (1955) Nutrition and growth of several blue-green
algae. Am J Bot 42:282
Lowry OH, Rosenbrough NJ, Farr AL, Randal RJ (1951) Proteins esti-
mation by folin phenol method. J Biol Chem 193:265–265
Mahalakshmi S, Lakshmi D, Menaga U (2015) Biodegradation of dif-
ferent concentration of dye (Congo red dye) by using green and
blue green algae. Int J Environ Res 9:735–744
Mansour HB, Houas I, Montassar F, Ghedira K, Barillier D, Mosrati R,
Chekir-Ghedira L (2012) Alteration of in vitro and acute in vivo
toxicity of textile dyeing wastewater after chemical and biological
remediation. Environ Sci Pollut Res 19:2634–2643
Mathias AC, Rilwan IM (2013) Effect of indigo dye effluent on the
growth, biomass production and phenotypic plasticity of Scened-
esmus quadricauda (Chlorococcales). An Acad Bras Cienc
86:19–428
Moosvi S, Keharia H, Madamwar D (2005) Decolorization of textile
dye Reactive violet 5 by a newly isolated bacterial consortium
RVM11.1.World. J Microbiol Biotechnol 21:667–672
Moran R (1982) Formulae for determination of chlorophyllous pig-
ments extracted with N, N-dimethylformamide. Plant Physiol
69:1376–1381
Naik DJ, Desai HH, Desai TN (2013) Characterization and treatment of
untreated wastewater generated from dyes and dye intermediates
manufacturing industries of Sachin industrial area, Gujarat, India.
J Environ Res Develop 7(4A):1602
OECD 201 (1984) OECD guideline for testing of chemicals. ‘alga,
growth inhibition test
Pinheiro HM, Touraud E, Thomas O (2004) Aromatic amines from azo
dye reduction: status review with emphasis on direct UV spec-
trophotometric detection in textile industry wastewaters. Dyes
Pigment 61:121–139
Przystaś W, Zabłocka-Godlewska E, Grabińska-Sota E (2012) Biologi-
cal removal of Azo and Triphenylmethane dyes and toxicity of
process by-products. Water Air Soil Pollut 223:1581–1592
Samchetshabam G, Hussan A, Choudhury TG (2017) Impact of textile
dyes waste on aquatic environments and its treatment. Environ
Ecology 35:2349–2353
Shukla SP, Kumar A, Tiwari DN, Mishra BP, Gupta GS (1992) Assess-
ment of the effect of the toxicity of a textile dye on Nostoc musco-
rum ISU, a diazotrophic cyanobacterium. Environ Pollut 84:23–25
Yaseen DA, Scholz M (2018a) Comparision of experimental ponds for
the treatment of dye wastewater under controlled and semi-natural
conditions. Env Sci Pollut Res 24:16031–16040
Yaseen DA, Scholz M (2018b) Treatment of synthetic textile waste-
water containing dye mixtures with microcosms. Env Sci Pollut
Res 25:1980–1997
13