Malays. Appl. Biol.

(2022) 51(3): 25-35

https://doi.org/10.55230/mabjournal.v51i3.2190

BIODEGRADATION OF METHYLENE BLUE BY BACTERIA

STRAINS ISOLATED FROM CONTAMINATED SOIL
FAZILAH ARIFFIN1,2* and NUR EQA MARDIHAH CHE ANUAR1

1
Faculty of Science and Marine Environment, Universiti Malaysia Terengganu,
21030 Kuala Nerus, Terengganu, Malaysia
2
Biological Security and Sustainability Research Group, Universiti Malaysia Terengganu,
21030 Kuala Nerus, Terengganu,Malaysia
*
E-mail: [email protected]

Accepted 17 June 2022, Published online 30 September 2022

ABSTRACT

Methylene blue is one of the textile dyes that are commonly used in the textile coloring industry. The discharge of textile
wastewater containing residual textile coloring substances into the environment can lead to environmental pollution. Thus,
bioremediation can be a solution to reduce dye pollution by using bacterial strains. In the present study, two bacterial strains with
the ability to degrade methylene blue dye were isolated from contaminated soil. Both isolated bacteria were further evaluated for
the dye decolorization percentage and the effect of abiotic parameters on bacterial growth. The isolated bacteria were incubated
in a mineral salt medium added with methylene blue dye for eight days in static aerobic conditions. The dye degradation was
examined by using UV-VIS spectrophotometer at 665 nm. The isolated bacteria were identified as Klebsiella pneumoniae strain
UMTFA1 (EK) and Pseudomonas aeruginosa strain UMTFA2 (EP) using 16s rRNA sequencing. The biodegradation study
showed that Klebsiella pneumoniae strain UMTFA1 (EK) and Pseudomonas aeruginosa strain UMTFA2 (EP) were capable to
degrade 10.52% and 11.65% of methylene blue dye after 8 days of incubation, respectively. The present study may provide a
basis for biotreatment and bioremediation of dye-contaminated soils.

Key words: Biodegradation, dye, methylene blue dye, soil bacteria

INTRODUCTION textile dyes into wastewater can lead to pollution.

Even at a concentration as low as 1 mg/L (1 ppm),
The textile manufacturing industry has been one of the dyes can easily be visible and detected by naked
the most popular sectors nowadays and is among eyes (Guaratini & Zanoni, 2000). According to
each country’s primary industries. In Malaysia, the Vimala et al. (2015), 50% of the textile effluent that
textile industry contributes to the Gross Domestic has been discharged contains a residual dye and it is
Product (GDP) by about 1.2% which is significantly challenging to be removed by conventional treatment.
important for the economy (Ślusarczyk et al., 2019). Thus, dye pollution will also affect groundwater due
Along with the textile industry’s popularity, dyes to soil leaching (Sodeinde & Eboreime, 2013). Textile
have played a major role as a textile colorant. It is dyes can persist in the environment and are difficult
expected that textile industries have accounted for the to degrade due to their complex structure (Sodeinde
extensive consumption of dyestuffs which is nearly & Eboreime, 2013). The biological transformation
80% (Vimala et al., 2015). Dyes exist in different of these dyes may produce aromatic amines that
structures that give them other names like azo, can be absorbed dermally and are known to have
reactive, disperse, basic, acidic, metal-complex, and genotoxic and carcinogenic properties (Brüschweiler
anthraquinone-based dye. According to Semeraro et & Merlot, 2017). This aromatic amine released during
al. (2015), more than 10,000 tons per year and around the breakdown of the dye linkages will turn the dye
100 tons/year of dyes are released into wastewater. colorless (Jaiswal & Gomashe, 2017). Usually, the
Besides being used as a colorant in textiles, these dyes biological decolorization of the dye is done under
are also widely used in the printing, pharmaceutical, anaerobic conditions. However, aromatic amines and
and food industries (Gayathri et al., 2014). other carcinogenic and mutagenic metabolites are still
Wastewater discharged from textile industries being released from the decolorized dyes (Kausar et
contains many pollutants including textile dyes al., 2016).
(Yaseen & Scholz, 2019). Improper discharge of Various physical methods (adsorption,
membrane filtration, ion exchange, and electrokinetic
* To whom correspondence should be addressed coagulation) and chemical methods (Fenton’s reagent,
26 BIODEGRADATION OF METHYLENE BLUE BY BACTERIAL STRAINS

ozonation, photochemical, and electrochemical by using bacteria to degrade the dye structure and
destruction) have been proposed to remove the dye used the compound for their energy requirement
from textile wastewater effluents. Though all of these and growth (Rumky et al., 2013). Thus, bacterial
methods can efficiently decolorize and remove the degradation of textile dye can help to mineralize
dye from wastewater, most of it is not economically and reduce the toxicity of the dye. Furthermore, this
feasible for larger-scale use, can generate a large method is low in cost, has lower sludge production,
amount of sludge, and need a high disposal cost and is environmentally friendly. Therefore, this study
(Robinson et al., 2001). As a viable alternative, aims to isolate and identify bacterial strains with the
microbial decolorization of textile dyes has gained ability to degrade textile dye, methylene blue. The
great interest because of their cost-effectiveness, less bacterial strain that possessed the ability to degrade
sludge production, environmentally friendly, and methylene blue dye will be selected for further study
yields non-toxic end products. to determine their dye degrading activity.
Recent studies have shown that various
microorganisms can decolorize a broad range of MATERIALS AND METHODS
textile dyes (Banat et al., 1996; Saratele et al., 2011).
Over the year, several microorganisms have been Preparation of the dye
tested to decolorize and mineralize the azo dye under The textile dye that used methylene blue
certain environmental conditions including fungi, (chemical formula: C16H18ClN3S3H2O, molecular
bacteria, yeasts, and algae (Pandey et al., 2007). weight: 373.9  g/moL, solubility in water: 40  g/L)
According to Banat et al. (1996), fungi decolorization obtained from brand Sigma- Aldrich. The dye is in a
results in slow processing and needs a longer time powder state and the concentration of the dye stock
for optimum decolorization. In addition, some fungi solution used in this experiment was 1 g/L.
only decolorize the dye color in the same species
but do not affect the chromophore center of the Soil sample collection
dye. Thus, fungal decolorization is limited in large- The soil sample was taken from Habibi Batik
scale decolorization (Chen et al., 2003). In contrast, House in Kampung Pak Tuyu, Kuala Nerus,
bacterial decolorization can achieve a higher degree Terengganu (GPS location 5.4156624,103.083565)
of color removal and complete mineralization of the as shown in Figure 1. The soil sample was collected
dye under optimum conditions (Saratele et al., 2011). near the effluent wastewater channel using a sterile
The previous study by Jaiswal and Gomashe spatula for about 10 cm depth from the soil surface.
(2017), stated that discharging colored effluent The sample was then placed in sterile plastic and
into the water bodies has become a great deal since stored at 4 °C. The temperature and pH of the soil
the dye effluent contains highly toxic material were measured.
that can be carcinogenic and mutagenic to living
organisms. In addition to the dye toxicity, pollution Isolation of dye degrading bacteria
of the colored effluent on the water surface can For the isolation of the bacteria, 10 g of soil
reduce light penetration into inner layers interrupting sample was mixed in 100 mL of nutrient broth added
the photosynthetic activities, lowering the oxygen with 50 mg/L methylene blue dye for 48 h at 37 °C.
content, and harming the aquatic life (Pandey et al., Serial dilution was performed after the incubation
2007; Abo-state et al., 2017). Thus, removing dye period to isolate the bacterial colony that decolorized
from wastewater has been a significant concern as the methylene blue dye. Then, 0.1 mL of the sample
they are generally challenging to degrade naturally was pipetted on the nutrient agar supplemented
and reductive cleavage of the dye structure can lead with 0.1 g/L of methylene blue and the spread plate
to the formation of aromatic amines that can be technique was applied using a sterile glass spreader.
carcinogenic to living organisms (Pandey et al., 2007). The bacterial colony with decolorizing ability was
To overcome the environmental pollution caused by observed after 48 h of incubation. The bacterial colony
textile dye contamination in the wastewater, textile that can decolorize dye has produced a clearance
dye degrading bacteria should display the ability to zone around the colony. The bacterial colony with a
completely decolorize and mineralize textile dyes. clearance zone was streaked on the nutrient agar plate
The bioremediation process uses the bacterial to obtain a single colony for the bacterial identification
strain that can help reduce dye toxicity and pollution and dye degrading assay. The pure bacterial isolates
into the environment. Microorganisms used in the were stored at 4 °C on a nutrient agar slant.
bioremediation process will feed on the aromatic
amines as their carbon sources (Alabdraba & Bayati, Identification of the bacterial isolates
2014). Throughout this study, biodegradation by The bacterial isolates were observed by their
bacteria can reduce the environmental pollution colony morphological characteristics and biochemical
caused by releasing textile dyes into wastewater. activities. Gram staining was performed then followed
Biodegradation is the chemical dissolution process
 BIODEGRADATION OF METHYLENE BLUE BY BACTERIAL STRAINS 27

Fig. 1. Sampling site, Kampung Pak Tuyu, Kuala Nerus, Terengganu (Google Maps, 2021)

by SIM (Hydrogen- Sulphide, Indole, Motility) test, different pH (pH 4, pH 7, & pH 8), dye concentrations
oxidase test, citrate utilization test, nitrate reduction (50 mg/L, 100 mg/L, 150 mg/L, 250 mg /L and 400
test, and triple sugar Iron test (Advanced Microbiology mg/L), and temperatures (28 °C, 37 °C, & 50 °C). The
Lab Manual, 2021). BBL Crystal Identification Kit screening process was conducted one factor at a time
from Becton Dickinson Microbiology Systems, (OFAT). It is performed by supplementing methylene
Cockeysville, Md., was also used to identify the blue dye in 10 mL of the mineral salt medium under
bacterial isolates. (MSM) preferred condition. The MSM contained the
Direct sequencing of 16S rRNA gene sequence following components: 1.8 g K2HPO4, 4.0 g NH4Cl,
was done by PCR-amplified 16S rDNA. The bacterial 0.2 g MgSO4.7H2O, 0.1 g NaCl, 0.01 g FeSO4.7H2O in
16S rDNA, full-length of 1.5 kb was amplified using 1 L of distilled water (Zajic & Supplisson, 1972). For
universal primers 27F and 1492R. The PCR was all treatments, the bacterial density was checked after
performed as follows: 1 cycle (94 °C for 2 min) 24 h of static incubation using a spectrophotometer
for initial denaturation; 25 cycles (98 °C for 10 s; at a wavelength of 600 nm. The bacterial culture
53 °C for 30 s; 68 °C for 1 min) for annealing and condition that gives the highest growth rate was used
extension of the amplified DNA. The PCR products for the dye degradation analysis.
were purified by the standard method and directly
sequenced with primers 785F and 907R using Dye degradation analysis
BigDye® Terminator v3.1 Cycle Sequencing Kit After the screening process was conducted on
(Applied Biosystems) according to the manufacturer’s the effect of abiotic parameters on bacterial growth,
protocol. The phylogenetic tree was constructed using the bacterial isolates from the stock culture were re-
the Neighbor-Joining method (Saitou & Nei, 1987). cultured in the enrichment culture consisting of 100
The evolutionary distances were computed using the mL of MSM and 400 mg/L methylene blue dye. A
Maximum Composite Likelihood method (Tamura et control experiment was conducted containing 100
al., 2004) and are in the units of the number of base mL MSM and 400 mg/L methylene blue dye without
substitutions per site. Evolutionary analyses were bacterial isolate. The mediums were incubated under
conducted in MEGA X (Kumar et al., 2018). static conditions for 8 days and the absorbance of each
medium was observed every 24 h of culture. For all
Effect of abiotic parameters on the bacterial treatments, 10 mL of the sample and control were
growth centrifuged at 5000 rpm for 10 min. The supernatants
Two types of bacterial strains with potential were analyzed using a spectrophotometer at a
decolorizing ability were further investigated under wavelength of 665 nm (Eslami et al., 2017). Each of
different abiotic parameters. Screening of the dye the experiments was carried out in triplicate.
degrading was performed to determine the best culture The calculation for percentage of dye degradation
condition for the bacteria to grow and decolorize the activity was calculated using the following formula
dye. The screening process will help to identify the (Zhuang et al., 2020):
bacteria’s ability to decolorize methylene blue dye
under suitable pH, concentration, and temperature.
Each of the bacterial isolates with potential dye
decolorizing ability was evaluated by testing the
ability of the selected bacteria to degrade dye at
28 BIODEGRADATION OF METHYLENE BLUE BY BACTERIAL STRAINS

RESULTS AND DISCUSSION Identification of isolated bacteria

The cell morphology magnification with
Isolation of dye degrading bacteria immersion oil shows that the isolated bacteria turn to
In this study, after the combination of 10 g of be Gram-negative with rod shape for both Isolate 1 and
contaminated soil with 100 mL of nutrient broth Isolate 2. These bacteria were undergone biochemical
added with 50 mg/L methylene blue (MB) dye, the tests to further identify the dye-degrading bacteria.
results revealed that the MB from blue color (Flask The biochemical tests for Isolate 1 indicated positive
A) was decolorized and formed in the Flask B (Figure results for lactose and carbohydrate fermentation
2). The medium showed color changed from dark which showed the ability to reduce nitrate. Negative
blue to brown after 48 h of incubation. Then, 0.1 results are shown for citrate fermentation, oxidase, and
mL of the sample was spread on a nutrient agar plate SIM (Sulphide-Indole-Motility) test. Additionally,
supplemented with methylene blue dye. The agar the result for BBL Crystal Kit proves that Isolate 1
medium has shown a clearance zone of dye around the is Klebsiella pneumoniae. Meanwhile, Isolate 2,
bacterial colonies, which proved their decolorization shows that the bacterial strain is a non-lactose and
abilities (Figure 3). two bacteria colonies showed carbohydrate fermenter. Moreover, it also gives
decolorization abilities named Isolate 1 and Isolate 2. negative results for the SIM test and nitrate reduction
These observations indicated that the isolates test. Positive results are shown in citrate utilization
bacteria were able to decolorize dye in static conditions and oxidase test. Hence, BBL Crystal Kit indicates
even during low oxygen. This support by Cao et al. that Isolate 2 is Pseudomonas aeruginosa as shown
(2019) was conducted decolorization of Direct Blue in Table 1.
2B by indigenous bacterial consortium experiments The identification result was further confirmed by
under static conditions. A study by Lade et al. (2015), 16S rRNA sequencing as shown in Figure 4 showed
stated that 99% decolorization of Trypan Blue dye the highest similarity with Klebsiella pneumoniae and
occurred under microaerophilic conditions within Pseudomonas aeruginosa and identified and named
hour using native microbial consortium and only 12% as Klebsiella pneumoniae strain UMTFA1 (EK) and
found decolorization under aerobic conditions. Pseudomonas aeruginosa strain UMTFA2 (EP). In

A B

Fig. 2. The color changes from dark blue (Flask A) to brown (Flask B) after 48 h of incubation.

(a) (b)

Fig. 3. The clearance zone for the dye decolorization on sample plate (b) compared to the control plate without soil microorganism
(a) after 48 h of incubation.
 BIODEGRADATION OF METHYLENE BLUE BY BACTERIAL STRAINS 29

previous studies, Klebsiella sp. was reported to have affects cell structures such as the membrane cell
azo dye-decolorizing abilities. The present study (Shah, 2013). A similar finding by Eslami et al.
shows similar to Holkar et al. (2018) which identified (2019), had shown that maximum decolorization
a facultative Klebsiella sp. C NCIM 5546 was based occurred at 37 °C. A similar effect of temperature (37
on the 16 S rDNA gene sequences and this bacterium °C) was observed in Malachite green dye degradation
was able to degrade reactive blue dye (Holkar et al. showed more than 97% by isolated soil bacteria under
2018). While a study done by Ariffin and Ariffin static conditions (Etezad & Sadeghi-Kiakhani, 2021).
(2020), proved that Pseudomonas aeruginosa based For the effect of concentration, the growth of
on the 16 S rDNA gene sequences was the dominant both isolates was tested in different concentrations in
bacteria for biodegradation of textile dye such as the range of 50 mg/L to 400 mg/L. Figure 5c shows
Reactive Green 19. that at the concentration of 400 mg/L, the bacterial
growth was the highest for both isolates. When higher
Effect of abiotic parameter on bacterial growth or lower concentrations of dye were provided in the
The effect of pH was varied at pH 4, pH 7, and enrichment culture, there were decreases in bacterial
pH 8. The highest number of bacteria growths was growth. Too little or too much initial concentration of
at pH 7 for both isolates after 48 h of incubation, as dye will not be able to induce the enzyme that can
shown in Figure 5a. It shows that the cells can tolerate break down the azo dye linkage because the enzyme
a range of pH because they do have some mechanisms is inductive (Dave et al., 2015). The present study
to regulate their cytoplasmic pH. However, there are showed the isolated bacteria growth was higher at a
limits to tolerance to their pH range. The bacterial dye concentration tested of 400mg/L. A similar finding
growth is decreased at pH out of this range due to by Roy et al. (2018), indicate that Enterobacter sp.
the destruction of the stability of plasma membrane, CV–S1 can decolorize and degrade a relatively higher
inhibition of membrane enzymes, and transport concentration of the crystal violet dye. Therefore, the
protein (Prescott et al., 2017) effect of dye concentration plays an important role in
Both isolates show the highest bacterial growth at the process of bioremediation of textile wastewater.
37 °C indicated in Figure 5b. Bacteria need optimum The effect of abiotic factors such as dye
temperature for growth. The higher temperature concentration, pH, temperature, time, glucose, and
causes thermal inactivation of proteins and probably sodium chloride concentrations on decolorization

Table 1. Bacterial identification results for Isolate 1 and Isolate 2

Isolate 1 Isolate 2
A. Morphological characteristics Colonies appear large, mucoid, and Colonies appear large, flat colonies
white on nutrient agar. with irregular margins and greenish on
nutrient agar.
Rod-shaped, Gram-negative
bacteria Rod-shaped, Gram-negative bacteria
B. Biochemical Test
• Mac Conkey Agar Pink colonies Colorless colonies
• Nitrate test Positive Negative
• Citrate utilization test Negative Positive
• Oxidase test Negative Positive
• SIM test H2S production: Negative H2S production: Negative
Motility: Negative Motility: Negative
Indole test: Negative Indole test: Negative
• Triple Sugar Iron test Slant: Yellow Slant: Red
Butt: Yellow Butt: Red
Gas production: Positive Gas production: Negative
H2S production: Negative H2S production: Negative
C. BBL Crystal Identification Kit Klebsiella pneumoniae Pseudomonas aeruginosa
D. Molecular Identification using Klebsiella pneumoniae strain Pseudomonas aeruginosa strain
16S rRNA sequencing UMTFA1 (EK) UMTFA2 (EP)
30 BIODEGRADATION OF METHYLENE BLUE BY BACTERIAL STRAINS

(a)

(b)
Fig. 4. Phylogram showed a genetic relationship between the isolated bacteria (a) Klebsiella pneumoniae strain UMTFA1 (EK)
(b) Pseudomonas aeruginosa strain UMTFA2 (EP) and other related reference bacteria based on the 16S rRNA gene sequence
analysis comparisons with accession numbers.

is important to determine the optimal conditions The present study showed that the decolorization
required for maximum decolorization or degradation rate for both isolates was found to be slower compared
of selected dye (Ikram et al., 2020). Results of previous to previous studies (Zabłocka-Godlewska et al., 2015;
studies have shown that the optimal conditions for the Garg et al., 2020). This might be due to the isolated
decolorizing dye activity of bacteria were pH 7, the bacteria utilizing only the dye in the MSM as a sole
temperature of 37 °C, the glucose concentration of source of carbon and energy without supplementing
6 g/L, and the dye concentration of 100 mg/L (Holkar any other carbon source. According to Mishra et al.
et al., 2018). (2020), the addition of some nutrients, such as carbon
sources (acetate and glucose) and nitrogen sources
Dye degradation analysis (yeast extract & peptone) enhanced the degradation
The study proceeds for the dye degradation process by the bacteria and maintain the diversity
activity by both isolates after eight days of incubation of the microbial community. The effect of abiotic
in static conditions. Figure 6 shows the decolorization parameters on the extensiveness of dye decolorization
percentage by Klebsiella pneumoniae strain UMTFA1 was tested by varying the carbon source and
(EK) and Pseudomonas aeruginosa strain UMTFA2 nitrogen source added to MSM and the maximum
(EP) incubated at 37 °C with a concentration of 400 decolorization was observed in MSM supplemented
mg/L and pH 7. The decolorization percentages for with peptone at pH 7 (Garg et al., 2020).
the Klebsiella pneumoniae strain UMTFA1 (EK) Another reason ineffectiveness of dye degradation
and Pseudomonas aeruginosa strain UMTFA2 (EP) could be considered is the mode of cultivation of the
are increasing from Day 0 to Day 8 with 10.52% and microorganism itself. Previous studies reported that
11.65% of decolorization percentages after 8 days, static conditions are preferred and are more efficient
respectively. The presence of this slight decolorization for azo dye decolorization (Prasad & Aikat, 2014).
may be correlated to the suitable temperature for the It is because the aromatic amines of its metabolite
enzymatic activity in many cells, which is normally products were assumed to resist further degradation
placed between 35 and 40 °C. However, the incubation compared with shaking cultivation conditions (Pandey
time may need to be extended further to see the effect et al., 2007). However, the effects of different modes
on the decomposition of the dye. of cultivation, either static or shaking conditions on
 BIODEGRADATION OF METHYLENE BLUE BY BACTERIAL STRAINS 31

dye decolorization efficiency, and the mechanisms In previous studies, many Klebsiella species were
of azoreductase activity and metabolite production reported to have azo dye-decolourizing abilities. For
during the dye degradation still need to be further example, Klebsiella strain Rz7 degraded 89.12 % of
investigated. Evans Blue dye (Zabłocka-Godlewska et al., 2015)
Several studies have shown that Pseudomonas while Klebsiella pneumoniae strain AHM completely
sp. exhibited high potential for selected dye decolorized 50 and 250 mg/L Reactive Orange dye
decolorization. Pseudomonas stutzeri, strain AK6 has 16 within 150 and 600 min under static condition
been studied and decolorized up to 86.2% within 96 h (Kumar et al., 2017). However, there are only a few
of Acid Blue 113 dye at 300 ppm (Hinsu et al., 2020). studies reporting methylene blue dye decolorization
While the findings reported by Garg et al. (2020) by Klebsiella pneumoniae.
showed that Pseudomonas aeruginosa decolorized
about 85% of 50 mg L−1 Reactive Yellow 145 dye CONCLUSION
within 4 days under static conditions. Pseudomonas
sp. SUK1 was approximately degraded by 25% of a In this study, two isolated bacteria from contaminated
mixture of ten azo dyes after 4 h and approximately soil were successfully identified known as Klebsiella
90 % after 24 h in static conditions (Zabłocka- pneumoniae strain UMTFA1 (EK) and Pseudomonas
Godlewska et al. 2015). aeruginosa strain UMTFA2 (EP). These isolated

(a) 0.7
0.6
0.5
0.4
OD(600nm)

Isolate 1
0.3 Isolate 2
0.2
0.1
0.0
pH 4 pH 7 pH 8

(b)
0.8
0.7
0.6
OD (600nm)

0.5
0.4 Isolate 1
0.3
Isolate 2
0.2
0.1
0
28˚C 37˚C 50˚C
Temperature (oC)

(c) 0.8
0.7
0.6
OD (600nm)

0.5
0.4 Isolate 1
0.3 Isolate 2
0.2
0.1
0
50 100 150 250 400
Dye Concentration (mg/L)

Fig. 5. Bacteria growth of Isolate 1 and Isolate 2 at different pH (a) different temperatures (b) different concentrations (c) after
48 h incubation in MSM supplemented with Methylene Blue dye.
32 BIODEGRADATION OF METHYLENE BLUE BY BACTERIAL STRAINS

(a)
12

10
% degradation

0
0 1 2 3 4 5 6 7 8

Incubation Time (Day)

Klebsiella
Klebsiella pneum oniae strain
pneumoniae strain UMTFA1
UMTFA1 (EK)
(EK) Control

(b)
14
12
10
% degradation

8
6
4
2
0
0 1 2 3 4 5 6 7 8

Incubation Time (Day)

Pseudomonasaeruginosa
Pseudomonas strain UMTFA2
aeruginosastrain UMTFA2 (EP)
(EP) Control

Fig. 6. Decolorization of methylene blue dye by Klebsiella pneumoniae strain UMTFA1 (EK) (a) Pseudomonas aeruginosa
strain UMTFA2 (EP) (b) after 8 days of incubation in MSM (at pH 7, 37 °C, and 400 mg/L concentration of dye).
 BIODEGRADATION OF METHYLENE BLUE BY BACTERIAL STRAINS 33

bacteria have the potential decolorizing 10.52% Blue 2B by indigenous bacterial consortium.
and 11.65% of methylene blue dye within 8 days. Journal of Environmental Management,
Therefore, it believes that these bacteria have the 242: 229-237. https://doi.org/10.1016/j.
potential in treating contaminated textile effluent jenvman.2019.04.067
as well as dye-contaminated soil. Further studies Chen, K.C., Wu, J.Y., Liou, D.J. & Hwang, S.C.J.
need to be carried out, such as factors that influence 2003. Decolorization of the textile dyes by newly
the performance of the bacteria to maximize the isolated bacterial strain. Journal of Biotechnology,
biodegradation process. 101: 57-68. https://doi.org/10.1016/S0168-
1656(02)00303-6
ACKNOWLEDGEMENTS Dave, S.R., Patel, T.L. & Tipre D.R.. 2015. Bacterial
degradation of azo dye-containing wastes. In:
This article is based on the work supported by the Microbial Degradation of Synthetic Dyes in
Ministry of Higher Education, Malaysia under the Wastewaters. S. Singh (Ed.). Springer, Cham. pp.
Fundamental Research Grant Scheme (FRGS 59519) 57-83. https://doi.org/10.1007/978-3-319-10942-
Grant No. FRGS/1/2018/STG05/UMT/02/3, the 8_3
Faculty of Science and Marine Environment and Eslami, H., Sedighi Khavidak, S., Salehi, F.,
Universiti Malaysia Terengganu (UMT) for research Khosravi, R., Fallahzadeh, R. A., Peirovi, R. &
facilities and support. Sadeghi, S. 2017. Biodegradation of methylene
blue from aqueous solution by bacteria isolated
CONFLICT OF INTEREST from contaminated soil. Journal of Advances in
Environmental Health Research, 5(1): 10-15.
The authors declare no conflict of interest Eslami, H., Shariatifar, A., Rafiee, E., Shiranian, M.,
Salehi, F., Hosseini, S.S. & Ebrahimi, A.A. 2019.
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