Bioscience Biotechnology Research Communications Vol 14 No (4) Oct-Nov-Dec (2021)

Toxicological Communication

On the Phytotoxicity of Waste-Water from Textile Industry

on Selected Crops Seed Germination and its Treatment
Using Bacteria and Zinc Oxide Nanoparticles

A. Nirmala,* S. Soundariya and A. Rajeshwari Praveena

Department of Biotechnology, Aarupadai Veedu Institute of Technology, Vinayaka
Missions Research Foundation, Paiyanoor, Chengalpatt, Tamil Nadu, India

ABSTRACT
Industrialization plays a major role for the economic development of any nation. In spite of various positive aspects of industrialization,
the foremost negative aspect is pollution by discharge of more waste water in to the environment. So, the aim of the present work
is to analyse the phytotoxic effect of textile industry waste water on seed germination and treating the waste water using Bacterial
Species and Zinc Oxide Nanoparticle. Physico-chemical parameters of dying industry waste water such as color, temperature, pH,
conductivity, turbidity, total dissolved solid, total hardness, COD, BOD, oil and greases, chloride, sodium, potassium, chromium,
copper, total alkalinity and zinc were analysed as per standard methods. To confirm the harmfulness, an investigation was made to
study the degree of toxicity of dyeing industry effluent on seed germination and growth of Sorghum bicolor (white sorghum) Vigna
unguiculata (cow pea) were selected for this study and it was placed in soil containing pots and watering with untreated industry
waste water and treated waste water. The waste water was treated by two methods, one by bacterial degradation, using Pseudomonas
fluroscence and the secondly, by chemical oxidation, using zinc oxide nanoparticles. After treatment their efficiency was tested with
above two plants. All the above said parameters were found to be high in untreated waste water. There was a gradual decrease in the
percentage of seed germination and seedling growth due to higher concentration of effluent, when compared with control. In this
comparative study, dye degradation(methyl orange) degradation by the chemical oxidation using zinc oxide nanoparticle was more
effective and faster than the biological oxidation of bacterial species.

KEY WORDS: Dye Degradation, Industrialization, Microorganism,

Nanoparticle, Phytotoxicity.

INTRODUCTION The waste water treatment system in Indian companies

was substantially established to fulfil the waste water
Water contamination is a current environmental concern discharge regulations, but only 10% of the waste water is
that has caught a lot of attention throughout the world. now treated, and the rest of the untreated water discharged
Textile dyeing and finishing industry effluents are the major into neighbouring water bodies. The major industrial
source of water contamination. The textile industry faces a pollutants are industrial effluents, which contain organic
challenge in using large quantities of dyes and auxiliaries that and inorganic chemicals, acids, alkalies, suspended
are required for current textile processes due to substantial particles, and other recalcitrant compounds (Mehta and
changes in client demands (Mamatha and Beena 2018). Bhardwaj 2012). However, dumping of untreated effluents
Factories discharged a wide range of pollutants, many of into the environment affects the ecological niches of living
which contain chemical elements in higher concentrations organisms (Bharagava et al. 2020). Municipalities from
to harm the surface and ground water supplies (Khan and government sector, even private entities face a serious of
Malik 2018; Bharagava et al. 2020). difficulty with wastewater disposal, particularly in large
urban areas with limited space for land-based treatment and
disposal. But still, they are taking corrective steps to make
safer (Naveenraj et al. 2021).
Article Information:*Corresponding Author: drnirmala81@gmail.com
Received 15/10/2021 Accepted after revision 17/12/2021 The textile mills daily discharging millions of litters of
Published: 31st December 2021 Pp- 1564-1570
untreated effluents loaded with synthetic dyes into public
This is an open access article under Creative Commons License,

1564
Published by Society for Science & Nature, Bhopal India.
Available at: https://bbrc.in/ DOI: http://dx.doi.org/10.21786/bbrc/14.4.30
Nirmala et al.,
wastewater stream that eventually drain into water bodies. Material and Methods
This can lead to acute toxicity of aquatic ecosystem.
This also impacts water quality by changing the pH and The effluent samples were collected from dyeing industry
increasing the Biochemical Oxygen Demand (BOD) and in plastic container located at Srivilluputhur, Tamil Nadu
Chemical Oxygen Demand (COD) (Imtiazuddin and (Fig-1 20L water sample). After collection, the effluent
Tiki 2020). Natural dyes come from a variety of sources, was immediately transported to the laboratory for analysis.
including vegetables, plants, minerals, insects, and animals. Physico-chemical parameters such as colour, temperature,
But synthetic dyes are made from organic molecules; pH, conductivity, turbidity, total dissolved solid, total
they may be produced in a consistent manner. These dyes hardness, COD, BOD, copper, chloride; chromium, sodium,
will linger in the environment for a long time if they are total alkalinity, potassium and zinc were analyzed as per
not properly treated (Olukanni et al. 2006). Among other the standard methods (APHA 2012).
dyes, Azo dyes are the greatest group of colorants in terms
of number and production volume, accounting for 60-70 The zinc oxide nanoparticle was obtained by the precipitation
percent of all organic dyes produced worldwide (Carliell et method. In a beaker 7.4g of zinc nitrate was taken in 50ml
al. 1998; Bafana et al. 2011; Imtiazuddin and Tiki 2020). of water. In another beaker 2.65g of Sodium carbonate was
taken in 50ml of water. Then sodium carbonate was added
The success of azo dyes is owing to their ease and low cost drop by drop to the solution of zinc nitrate. This mixture
of synthesis when compared to natural dyes, as well as their was stirred nearly 15-20min to get mixed together. Then
structural variety, high molar extinction coefficient, medium it was filtered using the Whatman filter paper to filter the
to high light and moisture fastness qualities. They're precipitate. The precipitate thus formed was washed with the
employed in a variety of industries, including textiles, water for 2 to 3 times to remove the minute particles. After
pharmaceuticals, and cosmetics, as well as food, paper, washing with water, the precipitate was again washed with
leather and paints industries (Chung et al. 1992; Mansour ethanol to remove the moisture present. Then it was dried
et al. 2007; Seesuriyachan et al. 2007). Azo dyes have an in a hot air oven for 90-100 degree for overnight period of
azo bond (–N=N–) in their basic structure, which is difficult 4-5 hrs. Thus the moisture was fully removed and obtained
to breakdown naturally. Depending on the number of azo a powdered substance. Finally powered substance was kept
groups, this class of dye is divided into mono azo, di azo, in a muffle furnace at 300℃ for 2hrs (Fig -2).
tris azo, and poly azo dyes (Engel et al. 2008). Reactive azo
dye is resistant due to their stability, xenobiotic nature, and For the characterization of zinc oxide nanoparticles, four
it is not completely destroyed by standard treatments such as main methods were considered.
light, chemicals, or activated sludge. (Grekova-Vasileva et • FT-IR (Fourier transform infrared spectroscopy)
al. 2009; Imtiazuddin and Tiki 2020). Hence in the present • XRD (x-ray diffraction)
study focused on treatment of textile industry waste water • SEM (scanning electron microscope)
containing Azo dyes by using bacteria and Nanoparticles. • UV-DRS (UV-Vis diffuse réflectance spectroscopy)

Waste water, on the other hand, is a resource that can be The methyl orange was degraded by using the zinc oxide
used for productive purposes because it includes nutrients nanoparticle. The zinc oxide nanoparticle was taken as a
that can be used in agriculture and aquaculture as well. As catalyst for the degradation process. 250 ml of textile dyeing
a result, wastewater can be viewed as a resource on one waste water was taken into a conical flask with the addition
hand and a problem on the other. The use of industrial of 0.5g catalyst zinc oxide nanoparticle. This was kept under
and domestic sewage effluents for irrigation has recently the sunlight for 24hrs and the stirring done after each 15min
become a popular method of repurposing waste water, which it undergoes with the photo catalytic reaction. The
owing to the availability of significant amounts of N, P, K, zinc oxide nanoparticle added to the waste water degrades
and Ca, as well as other critical nutrients (Niroula 2003, and decolorizes the compounds present in it. Then the
Ali et al 2020). absorbance of degraded waste water was measured by using
UV-visible spectrophotometer. The degradation of methyl
Its repurposing could have a positive impact on the rural orange was done by bacteria. Bacterial species was cultured
community. Another advantage of reusing wastewater for (Pesudomonas fluroscence) using minimal media. Then the
irrigation is water conservation; due to water shortage, cultured bacteria were added in to the waste water taken in a
industrial effluents are employed as a source of irrigation different concentration as 1, 2, 3, 4,5μl in 5ml. Thus it was
for crops; however, indiscriminate effluent uses, ignores the kept in an incubator for overnight and the absorbance was
fact that untreated effluent can have severe effects on crop measured for initial, 3rd and 6th day concentration. From
development and quality. Therefore, it is necessary to study the absorbance efficiency of degradation was noted.
the impact of these effluents on crop system before they
are recommended and used for irrigation (Hari et al. 1994; To study the effect of untreated and treated waste water on
Thamizhiniyan et al. 2009). The present investigation has seed germination and growth, the healthy and uniform seeds
been carried out to study the degree of toxicity of dyeing of Sorghum bicolor (White sorghum) and Vigna unguiculata
industry effluent on seed germination and development (cow pea) were selected and surface sterilization done with
of Sorghum bicolor (white sorghum) Vigna unguiculata 0.1% Hgcl2 and thoroughly washed with distilled water
(cow pea). These crops we are commonly cultivating in to avoid surface contamination. Germination experiments
agriculture land. were carried out (Fig-3) in two set of soil containing pots
(one pot for white sorghum and other pot for cow pea).

1565 Textile Industry Waste Water Treatment BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS
 Nirmala et al.,
One set irrigated with untreated waste water and another et al. 2010). The effluent selected for the present study was
set of pots irrigated with treated industrial waste water. The very dark brown to black color, with observed value 70
germination was observed and recorded after 48hours. hazen, it was higher than value of BIS (25 hazen). This value
indicates that the effluent was highly colored due to presence
Results and Discussion of different dyes, colour producing compounds, metals, pH
of the effluent, temperature during dyeing and bleaching of
It is important to study all features of the textile effluent fabrics. Color reduces aquatic life's photosynthetic activity
to improve environmental performance and also to sustain and has an impact on other metrics such as DO, BOD and
considerable quality of the individual companies. Below others. In general, traditional treatment methods have a hard
are the images of nanoparticles collected with the help of time to removing colours (Forgacs et al. 2004; Przystaś et
precipitation method and seed germination done to initiate al. 2012; Hubbe et al. 2012; Imtiazuddin and Tiki 2020).
the experiment.

The Physico- Chemical parameters of textile effluent have

Figure 3: Seed germination study
been analysed and the experimental results were compared
with standard BIS limits. The results of the analysis are
presented in table -1.

Effect of effluents on seed germination: Experiment on

Sorghum bicolor and Vigna unguiculata showed (Fig -4 a,
b) an inhibitory effect of untreated industrial effluents on
seed germination and its early growth.

Figure 1: Waste water collected from industry

Table 1. The Physico-chemical parameters of dyeing

industry effluent

Parameters Observed BIS

value Value

Colour , Hazen 70 25
Temperature 29.1 50
pH@25℃ 6.75 5.5-9
Conductivity, mS/cm 2822 300
Turbidity, NTU 266 10
Total dissolved solid , 3404 2,100
mg/ L
Waste water treated with Zinc oxide nanoparticle (Fig -5a, Total hardness , 188 500
b) and bacterial species (Fig -6a, b) used for irrigating
mg/L as CaC03
the seed containing pots (Sorghum bicolor and Vigna
Chemical oxygen 300 250
unguiculata).
demand, mg/L
Biological oxygen 128 100
Figure 2: Nanoparticle obtained from precipitation demand, mg/L
method Oil and greaces, mg/L 16 10
Chloride , mg/L as Cl 616 600
Sodium, mg/L 234 200
Potassium, mg/L 43.8 12
Chromium , µg/L BDL(DL1:0.5) 100
Copper , µg/L 0.047mg/L as cu 3000
Total alkalinity 138 mg/L as CaCO3
P-Alkalinity nil mg /l as CaCo3
Zinc, µg/L BDL(DL1:0.02) 5000
The colour of the effluent was key issue in textile industry, mg/l as Zn
and it was widely recognized as a important pollutant in
wastewater. In fact, even at low concentrations, colour in The waste water temperature observed to be 29.1℃. The
effluent is easily visible to human eyes, and no one likes the acidic and basic nature of the effluent can be identified by
appearance of unclean water (Ibrahim et al. 1996; Wijetunga pH value and also it determines the presence or absence of

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS Textile Industry Waste Water Treatment 1566
Nirmala et al.,

various ionic species of the textile effluent. The pH value et al. 2012). Turbidity is to measure the degree, in which
of the dyeing industry effluent was found to be little acidic the water loses its transparency due to the presence of
pH 6.75 but it was within the permissible limit prescribed suspended particulates. High intensity of scattered lights
by BIS (pH 5.5-9). Although no health-based guideline was it means that higher turbidity. The value of turbidity was
proposed for pH but sometimes, eye irritation and other skin recorded as 266 NTU which have been found to be much
disorders are associated with values of pH greater than 11, higher than the BIS limit (10 NTU). This may be due to
but in our result found within the BIS permissible limit. more colour, more total suspended solids and oily scum
Joshi and Kumar (2011) observed the pH of textile effluent incorporate with colloidal matter increases the turbidity
ranged from 7.6-7.9 whereas another study reported the pH which gives bad appearance and foul smell. Thus, turbidity
of dye industry effluent ranged between 8.2 and 9.0 (Joshi should be measured and treated carefully before disposal
and Kumar 2011; Ahmad et al. 2012; Farid et al. 2012; (Elango 2017; World Health Organization 2017; Bharagava
Imtiazuddin and Tiki 2020). et al. 2020).

Inorganic salts (bicarbonates, chlorides, and sulphates of

Figure 4: Seeds containing pots irrigated with untreated calcium, magnesium, potassium and sodium) and minor
waste water amount of soluble organic matter make up Total Dissolved
Solids (TDS). As a result, TDS is equal to the total number
of cations and anions in water. Although TDS is not
generally regarded a main pollutant, it is utilised as an
indicator of aesthetic features of drinking water as well as an
aggregate signal of the presence of a wide range of chemical
pollutants. The value of TDS ranges from 2320 to 9840,
indicating the presence of high amount of inorganic salts.
From this study we observed the TDS was about 3404 mg/L,
it was higher than the BIS value (2100mg/L) (Islam and
Guha 2013). The hardness of the analysed water sample was
188 mg/L as CaC03. The maximum limit of total hardness
recommended by WHO and American Public Health
Figure 5: Seed containing pot irrigated with Zinc oxide Association (APHA) are 500 and 250 mg/L respectively.
nanoparticle treated waste water Samples evaluated possess hardness within the permissible
limits according to APHA and WHO. Low hardness value
may be due to water softening steps carried out in textile
industries. Our findings are similar to study report published
in the past. (Prasad 2011; Kumari et al. 2011; Joshi and
Shrivastava 2012; Bharagava et al. 2020).

Chemical Oxygen Demand recorded in the effluent was 300

mglˉ1 exceeded the permissible limit prescribed by BIS
(250 mglˉ1). Saravanan et al. (2012) recorded that chemical
oxygen demand of tannery industry effluent was above the
acceptable limits. Increases in COD can be due to huge
Figure 6: Plants irrigated with Pseudomonas fluorescens amount of industrial wastes such as detergents, softeners,
treated waste water non-biodegradable dyeing chemicals, formaldehyde based
dye fixing agents etc. Higher concentration of COD in water
implies toxic conditions and the presence of biologically
resistant organic substances (Saravanan et al. 2012; Elango
2017; Bharagava et al. 2020).

The excess BOD due to the presence of more organic

contaminants of textile effluents in to a water body. The
low or nil BOD shows good quality water, whereas a high
BOD indicates the water is highly contaminated and not
suggested for drinking purposes. The experimental data
Electrical Conductivity is an important physical parameter of present work shows 128 mg/L BOD. It was found to
to measure the sodium hazard of water quality. The be higher than the permissible limit of BIS. Increased
Electrical Conductivity found in the effluent (2822 mS/ level of BOD leads to microbial oxygen demand causes
cm) was greater than that of the permissible limit of BIS reducing DO which may induce hypoxia conditions with
(300 mS/cm) this may be due to the continuous discharge subsequent adverse effects on aquatic biota (Goel 1997).
of the chemicals and salts used along with dyes in the Oil and grease value was recorded as 16 mg/L which was
industries. Our results in accordance with Saravanan et al. quite higher than BIS limits (10 mg/L). Presence of oil and
(2012) they reported that electrical conductivity of tannery grease in water bodies leads to the formation of oil layer,
industry effluent was above the accepted limits (Saravanan which causes significant environmental problems like it

1567 Textile Industry Waste Water Treatment BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS
 Nirmala et al.,

prevent the transfer of oxygen from atmosphere to water bacterial species used for irrigating the seed containing
medium as a result reduces the amount of dissolved oxygen pots (Sorghum bicolor and Vigna unguiculata). By this
(DO) at the bottom of the water bodies (Central Pollution comparative study of both processes, the dye degradation
Control Board 1990; Bharagava et al. 2020). by the chemical oxidation of zinc oxide nanoparticle was
more effective and faster than the biological oxidation of
In the present study the amount of chloride was recorded bacterial species (Fig -5a, b). The zinc oxide nanoparticle
as 616mg/L as Cl which exceeds the BIS limits (600 has an Advanced Oxidation Process (AOP). The zinc oxide
mg/L). Mainly high chloride content in water destroys nanoparticle also has a property of photo catalytic activity
the microorganisms which are necessary to maintain food thus it possesses the higher seed germination and growth,
chains in aquatic life and it was found to be favour the when compared with waste water treated with Pesudomonas
EC, TDS, TSS, alkalinity and sulphate (United States fluroscence (Fig – 6 a, b). The research finding of Hussein
Environmental Protection Agency 1999). The studies of (2013) reported that Zinc oxide was most effective catalyst
Sajani and Muthukkaruppan (2011) revealed that sugar with all types of dyes the extent of decolourization reaches
mills effluent contains high amount of chloride (United 100 %. The extent of decolourization was also 100 % when
States Environmental Protection Agency 1999; Samuel and anatase was used as photocatalyst but that need more time
Muthukkaruppan 2011). Sodium recorded in the effluent than zinc oxide was used (Falah Hussein 2013; Naveenraj
was 234 mglˉ1, this level was higher than the value set by et al. 2021).
BIS is 200 mglˉ. Potassium content recorded in the effluent
was 43.8 mg/l found to be higher than the permissible Conclusion
limit of BIS (12 mglˉ1). But other compounds like copper,
chromium, alkalinity and zinc levels are found to be trace The findings of present study suggest that the physico-
value (BDL). Our results are accordance with Falah and chemical parameters were relatively high in dyeing industry
Hussein (2013) who observed that no significant changes effluent and it was toxic to plants, severely affected the
were observed in the concentration of zinc, iron, cadmium, seed germination and seedling growth. The waste water
copper, nickel, cobalt and manganese before and after treated by chemical oxidation of zinc oxide nanoparticle
treatment of real and simulated textile industrial wastewater. was more effective and fast than the biological oxidation
Moreover, the concentrations of most of these metals were of a bacterial species in seed germination and growth. The
very low or non-detective (Falah Hussein 2013; Naveenraj study further concluded that the plant Sorghum bicolour was
et al. 2021). more suitable to grow when compare to Vulgna ungcuilata.
The future will concentrate on cost analysis of the treatment
Effect of effluents on seed germination: Experiment on techniques and also on treatment of complex dyes. In Tamil
Sorghum bicolor and Vigna unguiculata showed (Fig -4 a, Nadu, particularly in Tirupur, Erode, Karur, Srivilluputhur
b) an inhibitory effect of untreated industrial effluents on and other districts, the textile industries are growing very
seed germination and its early growth. Inhibition of seed fast due to its several advantages but on the other hand it is
germination may be due to greater number of dissolved one of the root causes for environmental pollution.
solids that increases the salinity and conductivity of the
absorbed solute by seed before germination moreover higher Acknowledgements
salt content also changes the osmotic potential outside the
seed thereby reducing the amount of water absorbed by the The study was financially supported by the Aarupadai
seed which results in retardation of seed germination (Mehta Veedu Institute of Technology, Vinayaka Missions Research
and Bhardwaj 2012; Naveenraj et al. 2021). Foundation and Presidency College, University of Madras,
Chennai.
Similar result reported by David Noel and Rajan (2015), was
made investigation to study the degree of toxicity of dyeing Conflict of Interests: Authors declare no conflict of
industry effluent on seed germination and early growth of interests to disclose.
Lady’s finger. Germination experiments were carried out
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