Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561

Contents lists available at ScienceDirect

Environmental Nanotechnology, Monitoring & Management

journal homepage: www.elsevier.com/locate/enmm

Removal of methyl orange from aqueous solution using SRB supported

Bio-Pd/Fe NPs
Hemamalathi Kubendiran a, Dayal Hui a, Mrudula Pulimi a, N. Chandrasekaran a,
P. Sriyutha Murthy b, Amitava Mukherjee a, *
a
Centre for Nanobiotechnology, Vellore Institute of Technology, Vellore, India
b
Water & Steam Chemistry Division, BARC Facilities, Indira Gandhi Centre for Atomic Research Campus, Kalpakkam 603 102, Tamil Nadu, India

A R T I C L E I N F O A B S T R A C T

Keywords: Removal efficacy of biogenically (Desulfovibrio desulfuricans) synthesized bimetallic Pd/Fe nanoparticles on the
Methyl orange toxic azo dye Methyl Orange (MO) was evaluated using DM and natural water bodies. Nanoparticle concen­
Pd/Fe nanoparticles tration and conditions were optimized using response surface methodology (RSM). Biogenic Pd/Fe nanoparticles
Desulfovibrio desulfuricans
exhibited higher removal (96.16 ± 1.6%) efficiency in just 30 min. The role played by D. desulfuricans and Pd
In-situ removal efficacy
catalyst in MO removal was also assessed and 36.8 ± 1.4% removal by D. desulfuricans and 45.19 ± 1.7% by the
Cell viability assessment
Bio-Pd NPs were noted. Mechanism of removal of MO by Bio-Pd/Fe NPs as revealed by XRD, FTIR and ORP
analyses showed both adsorption and reduction processes. Removal studies carried out with the lake, tap and
ground water samples showed efficiency of 74 ± 1.95%, 81 ± 2.57% and 87 ± 1.36% respectively. Bio-Pd/Fe
nanoparticle treated MO exhibited reduced toxicity towards algal and bacterial species. Biogenic synthesis of
Pd/Fe NP offers a promising approach for removal of toxic methyl orange dye from contaminated aquatic
ecosystems and a methodology for remediation of MO is elucidated.

1. Introduction
Methyl orange (MO), an azo dye is considered as the most primitive
pollutant in wastewater and natural water bodies due to their compli­
cated molecular structure, synthetic origin, toxicity, carcinogenicity and
visible colour. MO is one among the most common and extensively used
acidic/anionic dyes in textiles, printing, cosmetics, electroplating,
mining, leather, pharmaceutical, tanning paper industries (Ismail et al.,
2019; Mahato et al., 2020). Around 0.7 million tons of toxic dyes con­
taining highly hazardous chemicals and by-products are being generated
annually by textile industries alone (Kulandaivel et al., 2014; Sadeghi
et al., 2019). The existence of an azo group on methyl orange and low
biodegradability make it a matter of consideration for environmental
studies to remove/remediate it.
In a majority of cases unused and left-over synthetic dyes are
released into natural water bodies without proper treatment methods
(He et al., 2020; Sun et al., 2019). These dyes are heat-resistant, muta­
genic in nature and even small traces are highly toxic. These dyes cause
renal disturbance, dysfunction in the digestive tract system, cerebral
damage, hepatic system, skin and central nervous system. They also
minimize food intake ability and inhibit growth and fertility rate (Dotto
et al., 2013; Feng et al., 2012). Studies have revealed that the azo
reductase enzyme of liver has the ability to reduce the azo bond
(–N– – N–) and produce aromatic amines, which could lead to intestinal
cancer (Chung, 2016; Feng et al., 2012). In natural ecosystems these
dyes hinders the passage of sunlight through the water column thus
affecting photosynthesis which in-turn results in reduced dissolved ox­
ygen (DO) level in water bodies (Chequer et al., 2013; Hussain et al.,
2019). Hence removal of toxic dyes from aquatic systems is highly
essential.
A number of physico-chemical techniques like sol – gel, chemical
coprecipitation, electrochemical, ball friction, hydrothermal, micro-
emulsion, coagulation, flocculation, nanofiltration, photolysis, acti­
vated carbon adsorption, reverse osmosis and ion exchange had been
successfully investigated (Hassaan and El Nemr, 2017; Katheresan et al.,
2018). Most of these techniques are energy intensive and require
extensive handling processes and not eco-friendly (Saini, 2017; Unuofin
et al., 2019). Also, these methods are not advantageous as they only
transmit the pollutants from one phase to another and also high
equipment cost limits their large-scale implementation. Hence nano

* Corresponding author at: Centre for Nanobiotechnology, VIT, Vellore 632014, Tamil Nadu, India.
E-mail address: amitav@vit.ac.in (A. Mukherjee).

https://doi.org/10.1016/j.enmm.2021.100561
Received 10 May 2021; Received in revised form 19 July 2021; Accepted 22 August 2021
Available online 25 August 2021
2215-1532/© 2021 Elsevier B.V. All rights reserved.
H. Kubendiran et al.
biotechnology has found a great attention by overcoming the above
problems using nanoparticles (NPs) (Jun et al., 2019; Sharma et al.,
2017). In water treatment, NPs usage is developing rapidly (Dhillon
et al., 2012; Saratale et al., 2018).
In this study, Pd/Fe nanoparticles had been synthesized using the
sulfate reducing bacteria (SRB) Desulfovibrio desulfuricans (hereafter
referred as Bio-Pd/Fe NPs). SRB was chosen as a supporting material for
formation of bimetallic Pd/Fe NPs to remediate MO, because of its
ubiquitous presence in wastewater systems. The novelty of the current
work is combining the catalytic activity of Pd/Fe bimetallic particles
with the SRB system, which has not been attempted so far. This proposed
novel bio-nano catalyst has increased stability as well as improved
removal strategy and hence it could be more effective than the existing
water treatment methods. To the best of our knowledge there are no
prior studies in this direction on dye removal using bimetallic Pd/Fe
nanoparticles supported with SRB. The current approach significantly
removed MO and the removal conditions were optimized using
Response surface methodology (RSM). The possible underlying mecha­
nism was also studied. Further the MO removal experiments were per­
formed using natural water samples and also the toxic level of MO before
and after treatment with Bio-Pd/Fe NPs was assessed towards bacterial
strains such as E. coli and Pseudomonas aeruginosa and freshwater
microalgae Scenedesmus sp to assure the environmental safety of the
transformed/degraded MO product.
2. Materials and methods
2.1. Materials
All the chemicals used were of analytical grade. Ferrous sulfate
(FeSO4⋅7H2O) was obtained from Sisco Research Laboratories Pvt. Ltd.,
India. Potassium Hexachloropalladate (IV) (K2PdCl6) was purchased
from Sigma-Aldrich and Methyl orange
(C14H14N3NaO3S) was from Hi–Media Laboratories, Mumbai, India.
All solutions were prepared using deionised water. Desulfovibrio desul­
furicans (NCIM 2047) was obtained from National Collection of Indus­
trial Microorganisms (NCIM Pune, India). SRB was grown and
maintained in Baar’s medium (K2HPO4 – 0.05 g; NH4Cl – 0.1 g; CaSO4 –
0.2 g; MgSO4⋅7H2O – 0.2 g; Sodium lactate – 0.7 g; Fe
(NH4)2(SO4)2–0.05 g; distilled water – 100 ml and pH – 7 to 7.5) (Mathur
et al., 2017).
2.2. Biological synthesis of bimetallic Pd/Fe nanoparticles
The biological synthesis method was followed as per previous report
with minor modifications to the reported protocol (Martins et al., 2017).
Desulfovibrio desulfuricans was grown in postgate medium and incu­
bated for 7 days at 37 ◦ C in anaerobic condition. After incubation, it was
transferred into falcon tubes (50 ml) inside an anaerobic chamber and
then centrifuged (Remi C-24BL) at 7000 rpm for 10 min. The superna­
tant was removed and the pellet was interacted with 0.05 M Potassium
Hexachloropalladate (IV) aqueous solution (pH 2.0 using 0.01 M HNO3)
(Humphries et al., 2006) inside an anaerobic chamber. Then, 100 ml of
culture media was transferred to serum bottles (125 ml) in aliquots and
sealed with butyl rubber stoppers. To replace the nitrogen in head space,
cell suspension was supplemented with sodium pyruvate (25 mM)
(injected through syringe). After 1 h of interaction at room temperature,
0.1 M of ferrous sulfate was added and allowed to interact for another 1
h until the color of solution changes to black. The resultant solution
containing cell bound Bio-Pd/Fe NPs was then separated by centrifu­
gation at 7000 rpm for 15 min. Resultant Bio-Pd/Fe NPs was then
washed with water thrice and then with ethanol (99.9%) once and dried
in vacuum oven (70 ◦ C).
2
Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561
2.3. Characterization of bimetallic Pd/Fe nanoparticles
The crystal structure was analysed using X-Ray Diffraction (Bruker
Advanced D8, Germany) using CuKa radiation (1.54 A˚). The surface
functional groups were analysed using Fourier-transform infrared
spectroscopy (IR Affinity-1, Shimadzu, Japan). To analyse the particle
size and shape of bacteria and Pd/Fe loaded biomass, Field emission
scanning electron microscope (FESEM) (Hitachi S 4800 HR, Japan) was
used.
2.4. Removal experiments by RSM
An important statistical method for optimizing the experimental
conditions and for building the quadrating model to assess the statistical
relationship among the parameters was obtained using the Response
surface methodology (RSM) (Biswas et al., 2019; Ravikumar et al.,
2020a). Through this method, industrial remediation processes by
nanomaterials and various chemical reactions can be designed (Nasiri
et al., 2018). The removal of methyl orange using Bio-Pd/Fe NPs was
investigated. The three important experimental parameters for the
removal of methyl orange are: 1) NPs concentration 2) initial MO con­
centration, and 3) pH. To optimize the experimental parameters,
factorial Central composite design (CCD) was used (Cho and Zoh, 2007).
Several permutation combinatorial experiments were conducted to
check the influence of one parameter on removal efficacy while other
two parameters were kept constant. The statistical significance of the
removal study findings was analyzed using ANOVA one-way by Design
Expert software version-11 (Dalvand et al., 2016). The ANOVA analyses
signifies whether the statistical model fits well or not from the Adjusted
R2 and predicted R2 values. Also, with the help of desirability score,
maximum MO removal at optimized condition can be predicted. The MO
concentration (40–80 mg/L), pH of the solution (5–9) and nanoparticle
dosage (500–1000 mg/L) were varied for optimization. These specific
ranges of these significant parameters had a say on MO removal as re­
ported by earlier studies (Deb et al., 2019; Jia et al., 2019; Uddin and
Baig, 2019).
As Desulfovibrio desulfuricans is anaerobic, removal of MO by Bio-Pd/
Fe NPs was performed under anaerobic condition. All MO removal ex­
periments were done in triplicates and it was carried out in 125 ml
serum bottles with 100 ml working volume. The bottles were then
purged with N2 for 10 min and closed with butyl rubber stoppers and
kept in shaker at room temperature. Periodically at specific time in­
tervals (10, 20 and 30 min), 2 ml of sample was collected and imme­
diately centrifuged at 8000 rpm for 10 min. The residual concentration
of methyl orange was measured using UV–vis spectrophotometer
(UV1750, Shimadzu Corporation, Kyoto, Japan) at 464 nm. The removal
percentage was calculated using the following equation:
MO Removal(%) =
Co − Ce
X100
C0
where C0 – initial methyl orange concentration; Ce (mg/L) – concen­
trations of methyl orange at equilibrium.
2.5. Removal of methyl orange using in-situ water samples
After optimizing the experimental conditions for MO removal in
double deionized water, MO removal using in-situ water samples was
studied. Water samples like lake water (lat: 12◦ 58′ 10.3′′ N, long:
79◦ 09′ 36.5′′ E), tap water (lat: 12◦ 58′ 10.4′′ N, long: 79◦ 09′ 20.0′′ E) and
ground water (lat: 12◦ 57′ 55.5′′ N, long: 79◦ 09′ 15.5′′ ) were collected in
and near VIT campus and characterized using Inductively coupled
plasma – optical emission spectrometry [ICP-OES (Perkin Elmer Optima
5300 DV)] to check the trace elements present in these samples (Feist
and Sitko, 2019). Conductivity, pH of real water samples was also
checked. MO removal study in these natural water samples was
H. Kubendiran et al.
performed using Bio-Pd/Fe NPs at optimized conditions (Initial MO
concentration: 40 mg/L; Bio-Pd/Fe NPs concentration: 1000 mg/L; pH:
5).
2.6. Cell viability assessment to check the toxic level of MO before and
after treatment with Bio-Pd/Fe NPs
Bacterial strains such as E. coli and Pseudomonas aeruginosa and
freshwater microalgae Scenedesmus sp. were used as test organisms for
the toxicity assessment of MO (diluted to 1 mg/L) before and after
treatment with Bio-Pd/Fe NPs. For growing bacterial strains, nutrient
broth (Hi media, India) was used. During the exponential phase, test
bacterial strains were harvested through centrifugation at 6000 rpm for
10 min. In order to remove the residues of culture media ingredients, cell
pellet was washed with PBS (phosphate buffered saline). The bacterial
culture was prepared in nutrient broth and OD was adjusted to 0.1 (1 ×
108 CFU/mL) for the toxicity assessment studies. The untreated MO and
Bio-Pd/Fe NPs treated MO was added to 5 ml of bacterial suspension and
incubated at 37 ◦ C for 24 h as reported in previous study (Ravikumar
et al., 2019). The viability was tested using standard plate count assay
and the viability percentage was calculated using control as reference.
The control for this assessment study is bacterial culture devoid of any
treatment (taken as 100% viable).
The BG-11medium was used for growing freshwater microalgae
(Scenedesmus sp) with conditions: temperature-20 ◦ C; under white light
illumination (TLD Super 80 Linear fluorescent tube, Philips, India) of
intensity 3000 lx; day/night rhythm of 12/12 h. The culture was
centrifuged at 7000 rpm for 10 min during exponential growth phase
and the pellet was rinsed thoroughly with sterile fresh water and the OD
was adjusted to 0.5 (5 or 105 cells/mL). The pellet was then allowed to
interact with MO and Bio-Pd/Fe NPs treated MO for 72 h under visible
irradiation condition. The algal toxicity assessment was performed as
per the method reported previously (Roy et al., 2018). Number of intact
cells without any damage were counted using haemocytometer under
optical microscope. The viability percentage was calculated by keeping
control as reference. Culture without any treatment is kept as control
and they are taken as 100% viable.
3. Results and discussion
3.1. Characterization of bimetallic Pd/Fe nanoparticle
The obtained XRD results (Fig. 1) for Bio-Pd/Fe NPs showed Pd0
peaks at 2θ value of 40◦ and 68◦ which relates to cubic crystal planes
(1 1 1) (2 2 0) of metallic Pd [ICDD/JCPDS, PDF-# 65-2867] and Fe0
peak at 2θ value of 46.6◦ which relates to (2 0 0) reflective planes of
cubic phase of BCC iron [JCPDS no: 00-006-0696] (Khaghani et al.,
2017; Ravikumar et al., 2019; Suja et al., 2014; Wang et al., 2012).
Fig. 1. XRD analysis of Bio-Pd/Fe NPs.
3
Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561
The FT-IR analysis was used to check the changes in composition of
D. desulfuricans after adsorption of Pd. Fig. 2a represents the FT-IR
spectra of D. desulfuricans and Fig. 2b shows the spectra obtained for
Bio-Pd/Fe NPs. The FT-IR spectra of D. desulfuricans exhibited peaks at
3597 cm-1 represented N–H asymmetric stretching of the amines and
O–H bond of the hydroxyl groups (Kiran et al., 2017) and 2332 cm− 1
indicates the presence of C– – S group in the blank D. desulfuricans (Singh
et al., 2011). Main characteristic peaks at 1406, 1506 and 1633 cm− 1
correspond to the stretching vibration of benzenes in SRB as reported
previously (Ling-Li et al., 2012). For Bio-Pd/Fe NPs, peaks were found at
3259, 1633, 1506, 1081 and 817 cm− 1. New peaks at 1081 cm− 1 cor­
responds to –C–O–C stretching of Desulfovibrio strains (Feio et al.,
2004) and 817 cm− 1 corresponds to C–O–C and C–O–P stretching
due to presence of oligo and polysaccharides in the bacteria (Kiran et al.,
2017). As there was no much difference observed with the main char­
acteristic absorption peaks, indicating that the structure and composi­
tion of D. desulfuricans was not affected after absorption of Pd/Fe
nanoparticles on their surface.
The surface morphological characteristics and the elemental
composition of Bio-Pd/Fe NPs were analysed with the help of SEM and
EDAX. The biologically synthesized Pd/Fe nanoparticles using SRB is
represented in Fig. 3a and the average size of the Pd/Fe nanoparticles
formed was found to be 108 ± 2 nm (Fig. 3b) and the Fig. S1 (supple­
mentary information) represents the EDAX mapping and elemental
composition of Bio-Pd/Fe NPs. Previous reports also showed that the
size of microbial mediated NPs were approximately in range of 100–150
nm (Ahiwale et al., 2017; Srivastava and Mukhopadhyay, 2013). A
previous study reported that the size of the NPs become even lesser after
Fig. 2. FT-IR spectra of a) D. desulfuricans b) Bio-Pd/Fe NPs.
H. Kubendiran et al. Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561

Fig. 3. SEM image of a) D. desulfuricans b) Bio-Pd/Fe NPs.

addition of second metal (i.e., size of bimetallic NPs is lesser than the
Table 2
monometallic NPs) (Valiyeva et al., 2019). The implication of the
The analysis of variance (ANOVA) results for MO removal using Bio-Pd/Fe NPs.
smaller size of bimetallic NPs would result in increased specific surface
area and hence more reactivity. Source Sum of Squares df Mean Square F-value p-value

Model 2517.25 9 279.69 34.17 <0.0001

A-MO 904.61 1 904.61 110.52 <0.0001
3.2. Optimization of experimental conditions for MO removal by RSM B-NP 1112.40 1 1112.40 135.91 <0.0001
C-pH 43.23 1 43.23 5.28 0.0444
The 20 experimental runs for MO removal are given in Table 1 and AB 95.23 1 95.23 11.64 0.0066
AC 3.86 1 3.86 0.4719 0.5077
analysed using CCD. The MO removal efficiency by Bio-Pd/Fe NPs was BC 4.65 1 4.65 0.5685 0.4682
evaluated using ANOVA by Design expert (Version 11) and results are A2 192.24 1 192.24 23.49 0.0007
represented in Table 2. Based on analyses of experimental results, the B2 88.87 1 88.87 10.86 0.0081
following quadratic polynomial equation can be proposed: C2 37.30 1 37.30 4.56 0.0586
Residual 81.85 10 8.18
R1 = +29.68401–1.19356 MO + 0.118800 NP + 13.15354 pH-
Lack of Fit 68.51 5 13.70
0.000690 MO*NP + 0.034743 MO*pH + 0.003051 NP*pH + Pure Error 13.33 5 2.67
0.009131 MO2–0.000040 NP2–1.60877 pH2 Cor Total 2599.10 19
Where, MO is the methyl orange initial concentration (mg/L); NP is Std. Dev. 2.86 R2 0.9685
Bio-Pd/Fe NPs concentration (mg/L) and pH is the pH of the solution.
Mean 73.58 Adjusted R2 0.9402
The p-value (˃0.001) in Table 2 reveals the significance of the model
C.V.% 3.89 Predicted R2 0.7778
at a confidence level of 95%. The Predicted R2 of 0.7778 is in reasonable Adeq Precision 22.8478
agreement with the Adjusted R2 of 0.9402 (i.e. the difference is less than
0.2) and R2 value of 0.9685, this showed the excellent fit of the model.
Fig. S2 (supplementary information) shows the three-dimensional that the maximum removal of MO can be obtained at initial MO con­
(3D) surface contours of the predicted outcomes for MO removal. It centration of 40 mg/L, pH of 5 and nanoparticle concentration of 1000
was found that the removal percentage decreases when MO concentra­ mg/L. With the help of desirability score (0.985), the maximum removal
tion increases and removal percentage increases with increase in the of MO at optimized condition was calculated to be 95.76%. And
sorbent (Bio-Pd/Fe NPs) concentration and higher removal percentage experimentally, maximum MO removal at optimized condition was
at lower pH and these findings are in accordance with previous studies found to be 96.16%. Fig. S3 (Supplementary information) shows the
(Luo et al., 2016; Ravikumar et al., 2019). From these plots, it was found predicted Vs actual MO removal.

Table 1
3.3. Batch study- MO removal by Bio-Pd/Fe NPs
RSM experimental design and results for MO removal using Bio-Pd/Fe NPs.
3.3.1. Effect of Bio-Pd/Fe NPs concentration
Experimental Initial MO Bio-Pd/Fe NPs pH of the Results
runs conc. (mg/L) conc. (mg/L) solution
Adsorbent concentration is a significant factor which determines the
removal efficiency. The effects of various Bio-Pd/Fe concentration from
1 40 500 5 65.7
500 to 1000 mg/L on MO removal was evaluated by keeping initial MO,
2 60 750 6 68.84
3 60 750 4.31 70.14 pH and reaction time constant at 40 mg/L, pH 5 and 30 min respectively.
4 60 750 7.68 69.66 The results are depicted in Fig. S4A (Supplementary information). The
5 60 750 6 70.84 removal percentage was found to be 74.97 ± 1 0.7% for 500 mg/L;
6 60 329.55 6 50.64 81.92 ± 1.3% for 750 mg/L and 96.16 ± 1.6% for 1000 mg/L. It was
7 80 1000 5 71.07
8 60 1170.45 6 91.03
observed that the removal percentage increased with increase in Bio-Pd/
9 93.63 750 6 56.88 Fe concentration. The maximum removal percentage was obtained at
10 80 500 7 46.6 Bio-Pd/Fe NPs concentration of 1000 mg/L and hence the same is fixed
11 26.36 750 6 72.97 for further batch removal experiments. The percent of MO eliminated
12 40 1000 7 79.1
increased when the amount of Bio-Pd/Fe was raised, which can be
13 60 750 6 70.84
14 60 750 6 70.84 explained by the increased number of active sites available on the Bio-
15 60 750 6 70.84 Pd/Fe surface. As a result, the number of active connection sites
16 80 1000 7 74.97 participating in the adsorption process improved. These outcomes were
17 60 750 6 70.84 in accordance with previous studies (Ravikumar et al., 2016; Ravikumar
18 40 500 7 64.64
19 40 1000 5 98.24
et al., 2018).
20 80 500 5 45.5

4
H. Kubendiran et al.
3.3.2. Effect of MO initial concentration
The effect of three different MO initial concentration (40–80 mg/L)
was evaluated under Bio-Pd/Fe NPs concentration, pH and reaction time
constant at 1000 mg/L, pH 5 and 30 min respectively. From the results
(Fig. S4B (Supplementary information)), it is clear that the removal ef­
ficacy decreased with increase in MO initial concentration. The removal
percentage was found to be 96.16 ± 1.7% for 40 mg/L, and with further
increase in MO initial concentration, the removal percentage decreased
to 82.40 ± 1.5% for 60 mg/L and 71.07 ± 1.8% for 80 mg/L. The MO
initial concentration of 40 mg/L was chosen for further testing since it
resulted in higher removal. The decrease in the removal percentage with
increase in MO initial concentration is due the fact that the amount of
MO increased (40–80 mg/L) while the amount of absorbent remained
constant (1000 mg/L), thereby limiting the number of active adsorption
sites on the Bio-Pd/Fe surface. This can also be owing to competitive
adsorption among MO molecules on Bio-Pd/Fe NPs surface. Also due to
increased MO concentration, the active sites will be saturated faster and
thus there won’t be enough active sites on Bio-Pd/Fe NPs surface for MO
molecules to react with. These findings are similar to that of previous
studies (Murugesan et al., 2011; Yadav et al., 2012).
3.3.3. Effect of pH
The pH of the dye solution has a considerable effect on removal ef­
ficacy because it has influence on both the degree of ionisation of the dye
and the surface properties of the adsorbent (Van et al., 2018). The effect
of different pH (5–9) was evaluated by keeping Bio-Pd/Fe NPs concen­
tration, MO initial concentration and reaction time constant at 1000
mg/L, 40 mg/L and 30 min respectively. The results are depicted in
Fig. S4C (Supplementary information). The removal percentage was
96.16 ± 1.6% at pH-5; 92.56 ± 1.9% at pH-7 and 90.98 ± 0.6% at pH-9.
This removal process was slightly pH dependant because there was no
substantial variation in removal efficacy. Optimum pH for MO removal
was found to be pH-5 and the removal efficiency slightly decreased with
increase in pH. The mechanism can be explained as, due to adsorption of
anionic dye MO on positively charged Bio-Pd/Fe, formation of iron
oxides (i.e., corrosion) will be favoured and thereby release of more H +
will also occur at pH 5. The results suggested that electrostatic attraction
played a significant role in the adsorption mechanism of MO onto Bio-
Pd/Fe NPs. An earlier study shows that pH remained in the range of
5.8–6.2 at SRB and copper doped iron scrap enriched constructed
wetland microcosm at which maximum removal was observed (Yadav
et al., 2012). Another study revealed pH 5.3 is favourable for bio-nZVI/
Pd system to remove BDE-47 (Jia et al., 2019).
3.4. Comparative studies
At the above optimized conditions (sorbent weight-1000 mg/L; MO
initial concentration-40 mg/L; pH-5), 96.16 ± 1.6% MO removal was
observed for Bio-Pd/Fe NPs. The major crucial role was played by SRB in
MO removal process. SRB has the capability of oxidizing organic sub­
stances by using sulfate as an electron acceptor and as a result, biogenic
sulfide will be formed, which in turn can help the decolorization of azo
dye (Aullo et al., 2013; Yadav et al., 2012; Zhou et al., 2013). But the
limitation with SRB alone is delayed reaction time. Some studies show
that SRB can be supported with Fe0 NPs alone/bimetallic NPs for the
development of faster and more efficient sorbent (Guha and Bhargava,
2005; Zhou et al., 2013). A previous study witnessed that the cop­
per–iron bimetallic particles have positive influences on SRB activity
and also improves the removal of heavy metals (Zhou et al., 2013). Thus,
bimetallic Pd/Fe NPs were synthesized with SRB system in this study to
obtain enhanced MO removal. Other important advantage of using SRB
as a supporting material is, it prevents aggregation of Pd/Fe thereby
improves the stability and hence reactivity of Pd/Fe NPs will be more.
These are the reasons for excellent removal efficiency of Bio-Pd/Fe NPs.
Also, other advantage of using biologically synthesized bimetallic Pd/Fe
nanoparticles is that only lesser concentration of NPs will be utilized,
5
Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561
because majority of the weight will be biomass weight in 1000 mg/L
Bio-Pd/Fe NPs concentration.
At the above optimized condition, a control experiment was con­
ducted using SRB alone (without Pd/Fe) under anaerobic condition to
determine the role of SRB alone in MO removal process. The removal
percentage was observed to be 36.8 ± 1.4% under optimized conditions.
The initial MO removal by SRB would be through biosorption resulting
in dye removal, but later through the reductive cleavage of adsorbed MO
dye. Also, decoloration of azo dyes could happen through bio-mediated
chemical reduction of the azo dye by sulfide (H2S, HS–, S2–), an end
product of sulfate reduction by SRB. These outcomes are in accordance
with few previous studies (Rasool et al., 2016; Yadav et al., 2012). This
indicates that SRB plays crucial role in MO removal process by Bio-Pd/
Fe NPs.
To know the importance of Pd catalyst, control experiment was
conducted using Pd NPs supported SRB (Bio-Pd) under anaerobic con­
dition and at the optimized conditions. The removal percentage was
observed to be 45.19 ± 1.7% under optimized conditions. As compared
to removal percentage of SRB alone, Bio-Pd shows more removal effi­
cacy. This indicates that the removal efficacy of SRB was enhanced by
addition of Pd catalyst. Similar observations were reported in an earlier
study (Martins et al., 2017).
However, the removal efficacy of Bio-Pd can be further enhanced by
addition of Nanoscale zerovalent iron (Fe0–NPs) to this system. This is
because Fe0–NPs has gained prominence in environmental remediation,
particularly in the remediation of contaminants susceptible to reduced
transformation such as azo dyes (Tara et al., 2020; Zhao et al., 2020).
Their smaller particle size, larger specific surface area, higher density of
reactive surface sites and higher intrinsic reactivity of surface sites adds
its prominence in environmental remediation (Fan et al., 2019; Zhao
et al., 2020). Compared to conventional methods, nZVI can split the azo
bond and reduce dye molecules into products such as sulfanic acid, N, N-
dimethyl p-phenylene-diamine and Nethyl-p-phenylene-ediamine
(Cheng et al., 2013; Zhang et al., 2011). However, the decline in nZVI
reactivity over time is probably due to the formation of oxide layers onto
the particle surface during most of the reaction or when the nZVI par­
ticles come into contact with air (Wang et al., 2012; Wang et al., 2013).
To solve this problem, surface deposition of a second metal like Pd, Pt or
Ni can significantly increase the generation of available electrons to
reduce contaminants (Wang et al., 2012). Thus, in this study bimetallic
Pd/Fe NPs were synthesized on the surface of SRB to develop an efficient
sorbent material. From the MO removal results, it is confirmed that Bio-
Pd/Fe NPs are efficient sorbent material than SRB alone and Bio-Pd.
Table S1 (supplementary information) summarizes the other avail­
able nZVI nanocomposites, bacterial and yeast mediated removal of azo
dyes. As compared to removal by other nZVI nanocomposites, Biogenic
Pd/Fe NP from the current study has effectively removed higher con­
centration of MO and bacterial mediated removal takes longer reaction
time. The current removal study is a combination of both bacteria and
nZVI nanocomposites and hence can remove higher concentration of
MO in lesser reaction time (30 min).
3.5. Possible MO removal mechanism by biogenic-Pd/Fe nanoparticles
3.5.1. Changes in the MO solution during interaction
The chromophore group of MO dye exhibits the maximum visible
absorption wavelength at 464 nm (visible range), which is responsible
for the visible color of the dye. The presence of azo bond (N–
– N) is the
main cause for the visible colour of dye. During MO removal, changes in
absorption maxima of samples were observed (Fig. S5 (supplementary
information)). The absorption peak at 464 nm was found to be
completely decreased after 30 min of reaction time. This shows that the
dye has become colorless, thereby indicating that azo bond has been
cleaved by Bio-Pd/Fe NPs. Similar observations were obtained in earlier
studies (Chen et al., 2011; Han et al., 2015). Thus, MO removal mainly
occurs by degradation process.
H. Kubendiran et al.
During the 30 min interaction time of MO with Bio-Pd/Fe NPs, ORP
values were found to reduce from 315 to 169 mV (Fig. S6 (supplemen­
tary information)). This is a confirmation of a strong reducing envi­
ronment in the reaction mixture. The decrease in the ORP can be due to
the oxidation of Pd/Fe nanoparticles during interaction and thus indi­
cating that MO has been reduced. Previous studies show that during AB
113 dye removal by NZVI–resin, ORP values dropped indicating that AB
113 dye has reduced (Shu et al., 2010). Similarly, drop in ORP value was
observed during MO removal by HJ/nZVI indicating the prompt
oxidation of nZVI and reduction of dye (Li et al., 2017). For further
confirmation of oxidation of Bio-Pd/Fe NPs and reduction of MO,
detailed characterization was performed using XRD and FTIR.
3.5.2. Changes in Bio-Pd/Fe NPs after interaction
XRD spectra of Bio-Pd/Fe NPs before and after interaction with MO
are represented in Fig. 4. It was observed that after interaction with MO,
iron (II) oxide peaks at 2θ value of 34.68◦ and 37.62◦ and methyl orange
peaks at 2θ value of 21.78◦ and 43.58◦ were obtained as observed in
earlier study (Yahia et al., 2012). The presence of iron (II) oxide clearly
reveals that the Pd/Fe nanoparticles had been oxidized during interac­
tion, indicating that MO has been reduced. The presence of methyl or­
ange peaks shows that MO has been absorbed by Bio-Pd/Fe NPs. By this,
it is deduced that MO removal was found to occur by combination of
both adsorption and reduction. This outcome is supported by prior in­
vestigations, where MO removal occurred by combination of adsorption
Fig. 4. XRD analysis of a) Bio-Pd/Fe NPs (Before interaction) b) Bio-Pd/Fe NPs
(After interaction).
6
Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561
and reduction using Bio-nZVI (Ravikumar et al., 2018); similar way, acid
violet red B removal occurred by Fe NPs (Lin et al., 2014).
FT-IR results of MO and MO interacted Bio-Pd/Fe NPs are repre­
sented in Fig. 5a & 5b. The FT-IR spectra of MO dye exhibited peaks at
3420 (OH stretch vibration), 1600 (–N– – N– stretching of azo group),
1520 (–C– – C– stretching vibration), 1363 (confirms the sulphonic
nature of MO dye), 1201 (stretching vibrations of S– – O), 1118
(stretching vibration of C–O band), 955 (confirms Ring vibration) and
827 (stretching of distributed benzene rings) cm− 1 (Akansha et al.,
2019; Ayed et al., 2010; Chen et al., 2008; Uddin and Baig, 2019). After
interaction with MO, Bio-Pd/Fe NPs exhibited additional peaks at 3420,
1375, 1235, 1118, 955, 827 cm− 1, which represents the O–H bending,
C–O stretching, C–N stretching, C–H stretching vibrations of benzene
ring (Shen et al., 2015) of MO. The presence of these additional peaks
clearly indicate that MO had been adsorbed over the Bio-Pd/Fe NPs.
Similar results has been reported in previous study (Uddin and Baig,
2019). But, there was no 1600 cm− 1 peak (–N– – N– stretching of azo
group) found and this clearly indicates that azo bond of MO has been
cleaved by Bio-Pd/Fe NPs as in accordance with earlier study (Rasool
et al., 2013). From these outcomes, it can be concluded that MO removal
found to occur by combination of both adsorption and reduction.
3.6. Removal of methyl orange in real water samples
Table S2 (supplementary information) represents the trace elements
present in the real water samples as determined by ICP-OES. The pH of
lake water, tap water and ground water was found to be 7.2, 6.2 and 6.8
respectively and the conductivity was found to be 3.98 ± 0.5 mS cm− 1,
0.837 ± 0.5 mS cm− 1 and 0.977 ± 0.5 mS cm− 1 respectively. At opti­
mized condition, MO removal percentage for lake, tap and ground water
was found to be 74 ± 1.95%, 81 ± 2.57% and 87 ± 1.36% respectively
Fig. 5. FT-IR spectra of a) Methyl orange b) Bio-Pd/Fe NPs (After interaction).
H. Kubendiran et al.
Fig. 6. MO removal studies in real water samples.
(Fig. 6). These results suggested that the Bio-Pd/Fe NPs can be used for
the removal of MO in various environmental water matrices. But the
removal percentage for real water samples was found to be lower than
the removal efficiency of Bio-Pd/Fe NPs for deionized distilled water.
The possible reason would be presence of colloidal particles and trace
elements in the natural water samples (Table S2 (Supplementary in­
formation)). This same tendency was documented in few previous re­
ports where removal efficiency decreased due to interference of
impurities in the real water matrices (Ravikumar et al., 2020b; Zhang
et al., 2017).
3.7. Cell viability assessment
The cell viability of both E. coli and Pseudomonas sp. was represented
in Fig. 7. It can be observed that E. coli is much more sensitive than the
Pseudomonas sp. For both treatment groups, a significant difference in
cell viability was observed for both species with respect to control. The
cell viability percentage for untreated MO was found to be 37 ± 2% for
E. coli and 40 ± 3% for Pseudomonas sp. In case of Bio-Pd/Fe NPs treated
MO, cell viability percentage was 73 ± 4.5% for E. coli and 84 ± 2.8% for
Pseudomonas sp. It was observed that the cell viability for Bio-Pd/Fe NPs
treated MO was more as compared to that of untreated MO.
Similarly, the viability of fresh water algae Scenedesmus sp. was
assessed and cell viability percentage was found to be 63 ± 1.3% for
untreated MO and 86.8 ± 1.6% for Bio-Pd/Fe NPs treated MO (Fig. 7).
The cell viability assessment for MO towards both bacteria and algae
was found to be significantly increasing after treatment with Bio-Pd/Fe
NPs as compared to untreated MO. This demonstrates that after treat­
ment with Bio-Pd/Fe NPs, MO has transformed to less toxic form. Few
previous reports also documented that after decolorization/degrada­
tion, MO has transformed to less toxic form (Du et al., 2015; Kubendiran
et al., 2021).
4. Conclusion
Results of the present study demonstrate the preparation of Bio-Pd/
Fe NPs using SRB. It was found that Bio-Pd/Fe NPs have enhanced MO
removal efficiency. Using conditions optimized by RSM, 96.16%
removal was obtained for Bio-Pd/Fe NPs. Removal application in nat­
ural water samples was also studied and it was found to be 74 ± 1.95%,
81 ± 2.57% and 87 ± 1.36% for lake, tap and ground water respectively.
The cell viability assessment showed that, after interaction with Bio-Pd/
Fe NPs, the toxic level of MO has been reduced (63 to 37% in E. coli, 60
to 16% in Pseudomonas sp. and 37 to 14% in fresh water algae). Results
of this study demonstrate that Bio-Pd/Fe NPs were found to be a best
available remediating agent for methyl orange dyes, and offers a po­
tential for application in situ treatment of wastewater.
7
Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561
Fig. 7. Cell viability assessment to check the toxic level of MO.
CRediT authorship contribution statement
Hemamalathi Kubendiran: Investigation, Methodology, Visuali­
zation, Formal analysis, Writing – original draft. Dayal Hui: Method­
ology, Visualization. Mrudula Pulimi: Methodology, Formal analysis.
N. Chandrasekaran: Formal analysis, Writing – review & editing. P.
Sriyutha Murthy: Writing – review & editing. Amitava Mukherjee:
Conceptualization, Methodology, Supervision, Project administration,
Writing – review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.enmm.2021.100561.
References
Ahiwale, S.S., Bankar, A.V., Tagunde, S., Kapadnis, B.P., 2017. A bacteriophage mediated
gold nanoparticles synthesis and their anti-biofilm activity. Indian J. Microbiol. 57
(2), 188–194.
Akansha, K., Chakraborty, D., Sachan, S.G., 2019. Decolorization and degradation of
methyl orange by Bacillus stratosphericus SCA1007. Biocatal. Agric. Biotechnol. 18,
101044.
Aullo, T., Ranchou-Peyruse, A., Ollivier, B., Magot, M., 2013. Desulfotomaculum spp.
and related gram-positive sulfate-reducing bacteria in deep subsurface
environments. Front. Microbiol. 4, 362.
Ayed, L., Khelifi, E., Jannet, H.B., Miladi, H., Cheref, A., Achour, S., Bakhrouf, A., 2010.
Response surface methodology for decolorization of azo dye Methyl Orange by
bacterial consortium: produced enzymes and metabolites characterization. Chem.
Eng. J. 165 (1), 200–208.
Biswas, S., Bal, M., Behera, S.K., Sen, T.K., Meikap, B.C., 2019. Process optimization
study of Zn2+ adsorption on biochar-alginate composite adsorbent by response
surface methodology (RSM). Water 11 (2), 325.
Chen, Y.-P., Liu, S.-Y., Yu, H.-Q., Yin, H., Li, Q.-R., 2008. Radiation-induced degradation
of methyl orange in aqueous solutions. Chemosphere 72 (4), 532–536.
Chen, Z.-X., Jin, X.-Y., Chen, Z., Megharaj, M., Naidu, R., 2011. Removal of methyl
orange from aqueous solution using bentonite-supported nanoscale zero-valent iron.
J. Colloid Interface Sci. 363 (2), 601–607.
Cheng, Y., Lu, M., Jiao, C., Liu, H.-J., 2013. Preparation of stabilized nano zero-valent
iron particles via a rheological phase reaction method and their use in dye
decolourization. Environ. Technol. 34 (4), 445–451.
Chequer, F.D., de Oliveira, G.A.R., Ferraz, E.R.A., Cardoso, J.C., Zanoni, M.B., de
Oliveira, D.P., 2013. Textile dyes: dyeing process and environmental impact. Eco-
friendly Textile Dye. Finish. 6, 151–176.
Cho, I.-H., Zoh, K.-D., 2007. Photocatalytic degradation of azo dye (Reactive Red 120) in
TiO2/UV system: Optimization and modeling using a response surface methodology
(RSM) based on the central composite design. Dyes Pigm. 75 (3), 533–543.
H. Kubendiran et al.
Chung, K.-T., 2016. Azo dyes and human health: a review. J. Environ. Sci. Health, Part C
34 (4), 233–261.
Dalvand, A., Nabizadeh, R., Reza Ganjali, M., Khoobi, M., Nazmara, S., Hossein
Mahvi, A., 2016. Modeling of reactive Blue 19 azo dye removal from colored textile
wastewater using L-arginine-functionalized Fe3O4 nanoparticles: optimization,
reusability, kinetic and equilibrium studies. J. Magn. Magn. Mater. 404, 179–189.
Deb, A., Kanmani, M., Debnath, A., Bhowmik, K.L., Saha, B., 2019. Ultrasonic assisted
enhanced adsorption of methyl orange dye onto polyaniline impregnated zinc oxide
nanoparticles: kinetic, isotherm and optimization of process parameters. Ultrason.
Sonochem. 54, 290–301.
Dhillon, G.S., Kaur, S., Verma, M., Brar, S.K., 2012. Biopolymer-based nanomaterials:
potential applications in bioremediation of contaminated wastewaters and soils. In
Comprehensive Analytical Chemistry (Vol. 59, pp. 91-129): Elsevier.
Dotto, G.L., Vieira, M.L.G., Esquerdo, V.M., Pinto, L.A.A., 2013. Equilibrium and
thermodynamics of azo dyes biosorption onto Spirulina platensis. Braz. J. Chem.
Eng. 30 (1), 13–21.
Du, L.-N., Li, G., Zhao, Y.-H., Xu, H.-K., Wang, Y., Zhou, Y., Wang, L., 2015. Efficient
metabolism of the azo dye methyl orange by Aeromonas sp. strain DH-6:
characteristics and partial mechanism. Int. Biodeterior. Biodegrad. 105, 66–72.
Fan, Z., Zhang, Q., Gao, B., Li, M., Liu, C., Qiu, Y., 2019. Removal of hexavalent
chromium by biochar supported nZVI composite: batch and fixed-bed column
evaluations, mechanisms, and secondary contamination prevention. Chemosphere
217, 85–94.
Feio, M.J., Zinkevich, V., Beech, I.B., Llobet-Brossa, E., Eaton, P., Schmitt, J.,
Guezennec, J., 2004. Desulfovibrio alaskensis sp. nov., a sulphate-reducing
bacterium from a soured oil reservoir. Int. J. Syst. Evol. Microbiol. 54 (5),
1747–1752.
Feist, B., Sitko, R., 2019. Fast and sensitive determination of heavy metal ions as
batophenanthroline chelates in food and water samples after dispersive micro-solid
phase extraction using graphene oxide as sorbent. Microchem. J. 147, 30–36.
Feng, J., Cerniglia, C.E., Chen, H., 2012. Toxicological significance of azo dye
metabolism by human intestinal microbiota. Front. Biosc. (Elite edition) 4, 568.
Guha, S., Bhargava, P., 2005. Removal of chromium from synthetic plating waste by
zero-valent iron and sulfate-reducing bacteria. Water Environ. Res. 77 (4), 411–416.
Han, L., Xue, S., Zhao, S., Yan, J., Qian, L., Chen, M., Yao, M., 2015. Biochar supported
nanoscale iron particles for the efficient removal of methyl orange dye in aqueous
solutions. PLoS ONE 10 (7), e0132067.
Hassaan, M.A., El Nemr, A., 2017. Health and environmental impacts of dyes: mini
review. Am. J. Environ. Sci. Eng. 1 (3), 64–67.
He, C., Lin, H., Dai, L., Qiu, R., Tang, Y., Wang, Y., Duan, P.-G., Ok, Y.S., 2020. Waste
shrimp shell-derived hydrochar as an emergent material for methyl orange removal
in aqueous solutions. Environ. Int. 134, 105340.
Humphries, A.C., Mikheenko, I.P., Macaskie, L.E., 2006. Chromate reduction by
immobilized palladized sulfate-reducing bacteria. Biotechnol. Bioeng. 94 (1), 81–90.
Hussain, S., Khan, N., Gul, S., Khan, S., Khan, H., 2019. Contamination of water resources
by food dyes and its removal technologies. Water Chemistry. IntechOpen.
Ismail, M., Akhtar, K., Khan, M.I., Kamal, T., Khan, M.A., Asiri, A.M., Seo, J., Khan, S.B.,
2019. Pollution, toxicity and carcinogenicity of organic dyes and their catalytic bio-
remediation. Curr. Pharm. Des. 25 (34), 3645–3663.
Jia, Y., Hu, F., Lv, Y., Chen, Y., Hu, Y., 2019. Biomineralization of 2′ 2′ 4′ 4′ -
tetrabromodiphenyl ether in a pseudomonas putida and Fe/Pd nanoparticles
integrated system. Chemosphere 221, 301–313.
Jun, L.Y., Yon, L.S., Mubarak, N., Bing, C.H., Pan, S., Danquah, M.K., Khalid, M., 2019.
An overview of immobilized enzyme technologies for dye, phaenolic removal from
wastewater. J. Environ. Chem. Eng. 102961.
Katheresan, V., Kansedo, J., Lau, S.Y., 2018. Efficiency of various recent wastewater dye
removal methods: a review. J. Environ. Chem. Eng. 6 (4), 4676–4697.
Khaghani, S., Ghanbari, D., Khaghani, S., 2017. Green synthesis of iron oxide-palladium
nanocomposites by pepper extract and its application in removing of colored
pollutants from water. J. Nanostruct. 7 (3), 175–182.
Kiran, M.G., Pakshirajan, K., Das, G., 2017. Heavy metal removal from multicomponent
system by sulfate reducing bacteria: mechanism and cell surface characterization.
J. Hazard. Mater. 324, 62–70.
Kubendiran, H., Alex, S.A., Pulimi, M., Chandrasekaran, N., Nancharaiah, Y.,
Venugopalan, V., Mukherjee, A., 2021. Development of biogenic bimetallic Pd/Fe
nanoparticle–impregnated aerobic microbial granules with potential for dye
removal. J. Environ. Manage. 293, 112789.
Kulandaivel, S., Kaleeswari, P., Mohanapriya, P., 2014. Decolorization and adsorption of
dyes by consortium of bacteria with agriculture waste. Int. J. Curr. Microbiol. Appl.
Sci 3 (12), 865–882.
Li, X., Zhao, Y., Xi, B., Meng, X., Gong, B., Li, R., Peng, X., Liu, H., 2017. Decolorization of
methyl orange by a new clay-supported nanoscale zero-valent iron: synergetic effect,
efficiency optimization and mechanism. J. Environ. Sci. 52, 8–17.
Lin, Y., Chen, Z., Chen, Z., Megharaj, M., Naidu, R., 2014. Decoloration of acid violet red
B by bentonite-supported nanoscale zero-valent iron: reactivity, characterization,
kinetics and reaction pathway. Appl. Clay Sci. 93-94, 56–61.
Ling-Li, W., Wan-Hong, M., Shu-Lian, W., Yu, Z., Man-Ke, J., Rui-Ping, L., Ying-Ping, H.,
2012. The contrastive research in the photocatalytic activity of BiOBr synthesized by
different reactants. J. Nanomat., 2012.
Luo, F., Yang, D., Chen, Z., Megharaj, M., Naidu, R., 2016. One-step green synthesis of
bimetallic Fe/Pd nanoparticles used to degrade Orange II. J. Hazard. Mater. 303,
145–153.
Mahato, N., Sharma, K., Sinha, M., Baral, E.R., Koteswararao, R., Dhyani, A., Hwan
Cho, M., Cho, S., 2020. Bio-sorbents, industrially important chemicals and novel
materials from citrus processing waste as a sustainable and renewable Bioresource: a
review. J. Adv. Res. 23, 61–82.
8
Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561
Martins, M., Mourato, C., Sanches, S., Noronha, J.P., Crespo, M.T.B., Pereira, I.A.C.,
2017. Biogenic platinum and palladium nanoparticles as new catalysts for the
removal of pharmaceutical compounds. Water Res. 108, 160–168.
Mathur, A., Bhuvaneshwari, M., Babu, S., Chandrasekaran, N., Mukherjee, A., 2017. The
effect of TiO2 nanoparticles on sulfate-reducing bacteria and their consortium under
anaerobic conditions. J. Environ. Chem. Eng. 5 (4), 3741–3748.
Murugesan, K., Bokare, V., Jeon, J.-R., Kim, E.-J., Kim, J.-H., Chang, Y.-S., 2011. Effect of
Fe–Pd bimetallic nanoparticles on Sphingomonas sp. PH-07 and a nano-bio hybrid
process for triclosan degradation. Bioresour. Technol. 102 (10), 6019–6025.
Nasiri, R., Arsalani, N., Panahian, Y., 2018. One-pot synthesis of novel magnetic three-
dimensional graphene/chitosan/nickel ferrite nanocomposite for lead ions removal
from aqueous solution: RSM modelling design. J. Cleaner Prod. 201, 507–515.
Rasool, K., Shahzad, A., Lee, D.S., 2016. Exploring the potential of anaerobic sulfate
reduction process in treating sulfonated diazo dye: microbial community analysis
using bar-coded pyrosequencing. J. Hazard. Mater. 318, 641–649.
Rasool, K., Woo, S.H., Lee, D.S., 2013. Simultaneous removal of COD and Direct Red 80
in a mixed anaerobic sulfate-reducing bacteria culture. Chem. Eng. J. 223, 611–616.
Ravikumar, K., Kubendiran, H., Gupta, R., Gupta, A., Sharma, P., Alex, S.A.,
Mukherjee, A., 2020a. In-situ coating of Fe/Pd nanoparticles on sand and its
application for removal of tetracycline from aqueous solution. J. Water Process Eng.
36, 101400.
Ravikumar, K., Kubendiran, H., Ramesh, K., Rani, S., Mandal, T.K., Pulimi, M.,
Mukherjee, A., 2020b. Batch and column study on tetracycline removal using green
synthesized NiFe nanoparticles immobilized alginate beads. Environ. Technol.
Innovation 17, 100520.
Ravikumar, K.V.G., Kumar, D., Rajeshwari, A., Madhu, G.M., Mrudula, P.,
Chandrasekaran, N., Mukherjee, A., 2016. A comparative study with biologically and
chemically synthesized nZVI: applications in Cr (VI) removal and ecotoxicity
assessment using indigenous microorganisms from chromium-contaminated site.
Environ. Sci. Pollut. Res. 23 (3), 2613–2627.
Ravikumar, K., Santhosh, S., Sudakaran, S., Nancharaiah, Y.V., Mrudula, P.,
Chandrasekaran, N., Mukherjee, A., 2018. Biogenic nano zero valent iron (Bio-nZVI)
anaerobic granules for textile dye removal. J. Environ. Chem. Eng. 6 (2), 1683–1689.
Ravikumar, K.V.G., Sudakaran, S.V., Ravichandran, K., Pulimi, M., Natarajan, C.,
Mukherjee, A., 2019. Green synthesis of NiFe nano particles using Punica granatum
peel extract for tetracycline removal. J. Cleaner Prod. 210, 767–776.
Roy, B., Chandrasekaran, H., Palamadai Krishnan, S., Chandrasekaran, N.,
Mukherjee, A., 2018. UVА pre-irradiation to P25 titanium dioxide nanoparticles
enhanced its toxicity towards freshwater algae Scenedesmus obliquus. Environ. Sci.
Pollut. Res. 25 (17), 16729–16742.
Sadeghi, M., Forouzandeh, S., Nourmoradi, H., Heidari, M., Ahmadi, A., Jami, M.S.,
Abdizadeh, R., Mohammadi-Moghadam, F., 2019. /+//Biodecolorization of
Reactive Black5 and Reactive Red120 azo dyes using bacterial strains isolated from
dairy effluents. Int. J. Environ. Sci. Technol. 16 (7), 3615–3624.
Saini, R.D., 2017. Textile organic dyes: polluting effects and elimination methods from
textile waste water. Int. J. Chem. Eng. Res. 9, 975–6442.
Saratale, R.G., Saratale, G.D., Shin, H.S., Jacob, J.M., Pugazhendhi, A., Bhaisare, M.,
Kumar, G., 2018. New insights on the green synthesis of metallic nanoparticles using
plant and waste biomaterials: current knowledge, their agricultural and
environmental applications. Environ. Sci. Pollut. Res. 25 (11), 10164–10183.
Sharma, N., Bhatnagar, P., Chatterjee, S., John, P., Soni, I.P., 2017. Bio
nanotechnological intervention: a sustainable alternative to treat dye bearing waste
waters. Indian J. Pharm. Biol. Res. 5 (01), 17–24.
Shen, T., Jiang, C., Wang, C., Sun, J., Wang, X., Li, X., 2015. A TiO 2 modified
abiotic–biotic process for the degradation of the azo dye methyl orange. RSC Adv. 5
(72), 58704–58712.
Shu, H.-Y., Chang, M.-C., Chen, C.-C., Chen, P.-E., 2010. Using resin supported nano zero-
valent iron particles for decoloration of Acid Blue 113 azo dye solution. J. Hazard.
Mater. 184 (1-3), 499–505.
Singh, R., Kumar, A., Kirrolia, A., Kumar, R., Yadav, N., Bishnoi, N.R., Lohchab, R.K.,
2011. Removal of sulphate, COD and Cr (VI) in simulated and real wastewater by
sulphate reducing bacteria enrichment in small bioreactor and FTIR study.
Bioresour. Technol. 102 (2), 677–682.
Srivastava, N., Mukhopadhyay, M., 2013. Biosynthesis and structural characterization of
selenium nanoparticles mediated by Zooglea ramigera. Powder Technol. 244, 26–29.
Suja, E., Nancharaiah, Y.V., Venugopalan, V.P., 2014. Biogenic nanopalladium
production by self-immobilized granular biomass: application for contaminant
remediation. Water Res. 65, 395–401.
Sun, B., Yuan, Y., Li, H., Li, X., Zhang, C., Guo, F., Liu, X., Wang, K., Zhao, X.S., 2019.
Waste-cellulose-derived porous carbon adsorbents for methyl orange removal.
Chem. Eng. J. 371, 55–63.
Tara, N., Siddiqui, S.I., Rathi, G., Chaudhry, S.A., Inamuddin, Asiri, A.M., 2020. Nano-
engineered Adsorbent for the removal of dyes from water: a review. Curr. Anal.
Chem. 16 (1), 14–40.
Uddin, M.K., Baig, U., 2019. Synthesis of Co3O4 nanoparticles and their performance
towards methyl orange dye removal: characterisation, adsorption and response
surface methodology. J. Cleaner Prod. 211, 1141–1153.
Unuofin, J.O., Okoh, A.I., Nwodo, U.U., 2019. Aptitude of oxidative enzymes for
treatment of wastewater pollutants: a laccase perspective. Molecules 24 (11), 2064.
Valiyeva, G., Di Palma, L., Hajiyeva, S., Ramazanov, M., Hajiyeva, F., 2019. Synthesis
and stabilization of bimetallic Fe/Ni and Fe/Cu nanoparticles. Dimens. Syst., 3 2019,
14.
Van, H.T., Nguyen, T.M.P., Thao, V.T., Vu, X.H., Nguyen, T.V., Nguyen, L.H., 2018.
Applying activated carbon derived from coconut shell loaded by silver nanoparticles
to remove methylene blue in aqueous solution. Water Air Soil Pollut. 229 (12), 1–14.
H. Kubendiran et al.
Wang, H., Ji, S., Wang, W., Linkov, V., Pasupathi, S., Wang, R., 2012. Pt decorated PdFe/
C: extremely high electrocatalytic activity for methanol oxidation. Int. J.
Electrochem. Sci. 7, 3390–3398.
Wang, T., Su, J., Jin, X., Chen, Z., Megharaj, M., Naidu, R., 2013. Functional clay
supported bimetallic nZVI/Pd nanoparticles used for removal of methyl orange from
aqueous solution. J. Hazard. Mater. 262, 819–825.
Yadav, A.K., Jena, S., Acharya, B.C., Mishra, B.K., 2012. Removal of azo dye in
innovative constructed wetlands: influence of iron scrap and sulfate reducing
bacterial enrichment. Ecol. Eng. 49, 53–58.
Yahia, I.S., Abd El-sadek, M.S., Yakuphanoglu, F., 2012. Methyl orange (CI acid orange
52) as a new organic semiconductor: conduction mechanism and dielectrical
relaxation. Dyes Pigm. 93 (1-3), 1434–1440.
Environmental Nanotechnology, Monitoring & Management 16 (2021) 100561
Zhang, C., Zhu, Z., Zhang, H., Hu, Z., 2011. Rapid reductive degradation of azo dyes by a
unique structure of amorphous alloys. Chin. Sci. Bull. 56 (36), 3988–3992.
Zhang, Y., Jiao, Z., Hu, Y., Lv, S., Fan, H., Zeng, Y., Hu, J., Wang, M., 2017. Removal of
tetracycline and oxytetracycline from water by magnetic Fe 3 O 4@ graphene.
Environ. Sci. Pollut. Res. 24 (3), 2987–2995.
Zhao, Y., Zhao, Z., Song, X., Jiang, X., Wang, Y., Cao, X., Si, Z., Pan, F., 2020. Effects of
nZVI dosing on the improvement in the contaminant removal performance of
constructed wetlands under the dye stress. Sci. Total Environ. 703, 134789.
Zhou, Q., Chen, Y., Yang, M., Li, W., Deng, L., 2013. Enhanced bioremediation of heavy
metal from effluent by sulfate-reducing bacteria with copper–iron bimetallic
particles support. Bioresour. Technol. 136, 413–417.