Journal of Advances in Biology & Biotechnology

Volume 27, Issue 9, Page 1152-1161, 2024; Article no.JABB.123137

ISSN: 2394-1081

Green Synthesis of Nanomaterials with

Phytochemicals for Treating Multidrug
Resistant Bacteria
Vadlamudi Nikhil a, Ayla Sanjay a,
Mohammad Aftab Khizer a,b, Mohd Asif a,b,
Syed Shah MinAllah Alvi b and Chand Pasha a*
a Department of Microbiology, Nizam College, Osmania University, Hyderabad, Telangana, India.
b Department of Microbiology, Mumtaz degree and PG College, Malakpet, Affiliated to Osmania

University, Hyderabad, Telangana, India.

Authors’ contributions

This work was carried out in collaboration among all authors. All authors read and approved the final
manuscript.

Article Information
DOI: https://doi.org/10.9734/jabb/2024/v27i91386

Open Peer Review History:

This journal follows the Advanced Open Peer Review policy. Identity of the Reviewers, Editor(s) and additional Reviewers,
peer review comments, different versions of the manuscript, comments of the editors, etc are available here:
https://www.sdiarticle5.com/review-history/123137

Received: 02/07/2024
Original Research Article Accepted: 04/09/2024
Published: 05/09/2024

ABSTRACT

The Bacteria with Multidrug resistance and Extreme drug resistance are increasing at a rapid rate.
Various methods have been employed to combat drug resistant bacteria. Major classes of
antibiotics aren’t effective against these bacteria. Alternative methods have been studied in recent
years. Nanoparticles are used against multidrug resistant bacteria; The green synthesized
nanoparticles are more reliable due to more shelf life and lesser toxicity relative to chemically
synthesized nanoparticles. Multi drug resistant E. coli and Staphylococcus aureus was isolated
from sewage samples. Green synthesized nanoparticles from various plants samples have been
_____________________________________________________________________________________________________

*Corresponding author: E-mail: cpasha21@gmail.com;

Cite as: Nikhil, Vadlamudi, Ayla Sanjay, Mohammad Aftab Khizer, Mohd Asif, Syed Shah MinAllah Alvi, and Chand Pasha.
2024. “Green Synthesis of Nanomaterials With Phytochemicals for Treating Multidrug Resistant Bacteria”. Journal of Advances
in Biology & Biotechnology 27 (9):1152-61. https://doi.org/10.9734/jabb/2024/v27i91386.
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

prepared with Zinc and Copper forming respective oxides with Neem, Nakara, Jatropha, Mango,
Clove, Ginger, Cardamom, Cinnamon and Betel against multidrug resistant Escherichia coli and
Staphylococcus aureus. Isolated E. coli was susceptible to Fluoroquinolone and Augmentin
whereas S. aureus was susceptible to vancomycin. Green synthesized nanoparticles had more
antimicrobial activity against E. coli and S. aureus than chemically synthesized nanoparticles and
plant extracts. Green synthesized Nakara CuO nano particles had inhibition zone of 31 ±0.6mm
and 30 ±0.7mm for E. coli and S. aureus respectively, ZnO nano particles of Nakara had 25
±0.6mm inhibition zone for E. coli and S. aureus. Green synthesized Jatropha CuO nano particles
had inhibition zone of 26 ±0.5 and 26 ±0.4mm for E. coli and S. aureus. ZnO nano particles of
Jatropha had 31±0.7mm and 30±0.7mm inhibition zone for E. coli and S. aureus respectively. The
Scanning electron microscopy studies revealed 26nm Jatropha ZnO and 25nm Nakara CuO
nanoparticles. Nano materials were found to be non-toxic in cell line studies. The present study
concludes to study the impact of green synthesized nanoparticles as an alternative to antibiotics to
combat multidrug resistant bacteria.

Keywords: Phytochemicals; surface coating; green synthesis; resistant bacteria; metal oxide
nanomaterials.

1. INTRODUCTION chemical approaches [6]. Metal and metal oxide

nanoparticles formed by green synthesis method
Excessive use of antibiotics by human, as well as are increasingly applied in the biomedical field,
in agriculture and in aquaculture led to the rapid including disease treatment, wound healing,
increase in multidrug resistant bacterial strains. immunotherapy, dentistry etc [7].
IDSA (The Infectious Diseases Society of
America) classifies antimicrobial resistance as It is planned in study to green synthesize nano
one of the major significant global threats to materials and evaluate the antimicrobial activity
human health [1]. Bacteria found resistant to against the extreme drug-resistant gram negative
majority of the known Antibiotics including those and gram-positive bacteria.
considered last-line treatments like vancomycin
[2]. Due to development of resistance to several 2. MATERIALS AND METHODS
antibiotics, antimicrobial medicines lose their Sample collection: 15 Sewage sample collected
effectiveness, making the infections harder or from different locations of Hyderabad Telangana,
impossible to treat, raising the risks of disease India during May, June 2024.
transmission, chronic illness and mortality.
2.1 Isolation of Escherichia coli and
Phytochemicals are active compounds naturally
occurring in plants, recognized for their potential Staphylococcus aureus
health benefits and contributions to human
nutrition and medicine. They play major role in Pure E. coli and S. aureus were isolated from
plant growth and defence mechanism against collected sewage samples. The samples were
competitors, predators and pathogens. spread on the MacConkey agar plate and
Numerous plants serve as significant sources of Mannitol Salt Agar (HIMEDIA, India) incubated at
antimicrobial complexes that demonstrate potent 37oC for 24hrs. Colonies which were similar
activity against bacterial strains. More than 7000 based on growth on specific media to E. coli and
species of wild consumable plants contribute S. aureus were sub-cultured on nutrient agar
nutrition in human being [3]. and most of the plates.
antimicrobial activity is yet to be studied [4,5]. 2.1.1 Characterization
Green synthesis of nanoparticles is carried out Colony morphology: Colony characteristics of
by producing nano particles through living cells the isolates were observed on specific media E.
through biological pathways, this method of coli on MacConkey Agar, S. aureus on Mannitol
synthesis is more efficient and higher yield Salt Agar.
compared to other methods. Green synthesis
methods are recognized for their eco-friendly Microscopy: Gram staining and microscopic
nature, non-toxicity, cost efficiency, and superior observations were performed to confirm the
stability compared to alternative, physical, and organisms isolated.

1153
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

2.1.2 Biochemical test Extraction of phytochemicals: Aqueous and

ethanolic phytochemical extracts of Neem,
Several biochemical tests were carried out as Nakara, Jatropha milk, Mango seed, Clove,
Indole test, Methyl Red test, Voges Proskauer Ginger, Cardamom, Cinnamon and Betel leaves.
test, Citrate utilization test, Catalase test and were performed. Similar extracts were also
Coagulase test were performed. prepared for spices.

2.2 Antibiotic susceptibility Test Aqueous extracts were prepared by suspending

10 grams of samples in 90ml of phosphate buffer
The Antibiotic susceptibility test of E. coli and S. and grinded. The mixture was heated at 60oC for
aureus were carried out by placing HIMEDIA 10mins in water bath. The collected solution was
antibiotic disc on Mueller Hinton agar consisting filtered through Whatman filter paper and filtrate
of Antibiotics: Ampicillin Methicillin, was collected in sterile screw cap bottles and
Cephalosporin, Tetracycline, Monobactam, stored at 4oC for further use.
Carbapenem, Sulfonamide, Nitroimidazole,
Ethanolic extracts were prepared by suspending
Macrolide, Rifamycin, Fluoroquinolone,
10 grams of samples in 90ml of ethanol in sterile
Elfamycin, Ceftazidime, Cefepime, Norfloxacin,
screw cap bottles, heated at 60oC for 10mins,
Levofloxacin, Chloramphenicol, Streptomycin,
after cooling ethanolic fraction was separated.
Augmentin, Kanamycin, Pencillin – G,
The filtrate was evaporated on rotatory
Gentamycin and Vancomycin.
evaporator at 65oC until the ethanol is
evaporated. The resultant powder was
Augmentin antibiotic solution was prepared by
suspended in water, filtered through Whatman
adding 20mg Amoxicillin in 10mg potassium
filter paper and stored at 4oC for further use.
clavulanate and the solution was added in wells.
2.4 Synthesis of Nano Particles
2.3 Molecular Analysis
Copper oxide and Zinc oxide nano particles were
16srRNA method was used to identify the prepared by taking 0.02M of copper sulphate and
bacterial cultures. Bacterial cultures were grown Zinc acetate separately, dissolving each in 100ml
on nutrient broth for overnight to isolate the water. The solutions were titrated against 1M
genomic DNA of E. coli and S. aureus by NaOH dropwise 40ml for 10mins at 60oC. Further
QIAamp DNA kits® Bacterial Genomic DNA stirring at 60oC without NaOH was done until
Purification Kit (QIAGEN). brick red and white colour precipitate was
observed for Copper and Zinc respectively
Extracted genome was amplified by Polymerase indicating the formation of copper oxide and Zinc
Chain Reaction. The universal primers with 16S oxide nano particles.
rRNA gene, forward primer (5′-
AGAGTTTGATCMTGGCTCAG-3′) and reverse Green synthesis of Nano particles was
primer (5′-CTGCTGCSYCCCGTAG-3′) were performed by taking 3.1 grams of copper
used for the amplification of the 16S rRNA gene sulphate and 3.6 grams of Zinc acetate was
fragment. The amplified PCR products were taken separately and suspended in 100ml of
sequenced by ABI DNA sequencer (Applied Aqueous extracts samples. The solutions were
Biosystem Inc). titrated against 1M NaOH dropwise 4ml for
10mins at 60oC. Further stirring at 60oC without
The computation analysis of 16s rRNA gene NaOH was done until brick red and white colour
sequence of E. coli and S. aureus isolates precipitate was observed for Copper and Zinc
were compared with sequences in National respectively indicating the formation of green
Center for Biotechnology Information (NCBI) by synthesized copper oxide and Zinc oxide nano
BLAST [8]. Phylogenetic trees were made to particles coated with aqueous extracts.
understand the evolutionary relationships by
2.5 Purification of Nano Particles
ClustalW [9].
Precipitated nanoparticles were collected in an
Selection of plant: The plant samples were Eppendorf and subjected to centrifugation at
collected from Anantagiri Hills forest, Vikarabad, 10,000rpm for 5mins. The pellet was collected
Telangana, India specimens were identified by and washed with non-ionized distilled water. The
Dr. Vijaybhaskar Reddy, Taxanomist, Department washed pellet was dried in hot air oven for
of Botany Osmania university, Hyderabad. overnight at 80oC.

1154
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

Table 1. Plant Sample collection and part of samples

Samples Scientific name Part of Sample

Neem Azadirachta indica Leaf
Nakara Ximenia americana Latex
Jatropha Jatropha curcas Latex
Mango Mangifera indica Kernel
Clove Syzygium aromaticum Clove flower buds
Ginger Zingiber officinale Rhizome
Cardamom Elettaria cardamomum Seeds
Cinnamon Cinnamomum verum Bark
Betel Piper betle Leaf

Characterization of Nano particles: Nano tetrazolium crystal. Absorbance was measured at

particles characterization was carried out by the 570 nm using a microplate reader (Bio-Rad,
Scanning electron microscopy and antimicrobial Hercules, CA, USA). The data were analyzed
activity. with 3 parallel experiments and were expressed
as mean ± standard deviation.
The antimicrobial activity of phytochemicals,
nano particles and green synthesized nano Statistical Analysis: Experiments were
particles was tested against E. coli and S. repeated thrice in triplicate (n=9) and value with
aureus. The 100µl cultures E. coli and S. aureus standard deviation is presented.
were spread on Mueller Hinton agar separately.
The 30µl liquid samples and solid samples of 3. RESULTS
3mg were added in wells and incubated at 37oC
for 24hrs. Colony on MacConkey agar morphology: E.
coli strain had small round, smooth and pink
Green synthesized CuO and ZnO nano particles
colonies with depression in middle.
were further characterized by Scanning electron
Staphylococcus aureus had round, convex
microscopy. The samples were sterilized under
colonies with yellow colonies on MSA agar due to
UV light and placed on SEM stubs, the samples
fermentation of mannitol.
were then gold coated and scanning electron
microscopy was performed.
Microscopic observations: E. coli was rod
2.6 Toxicity shaped, gram negative, non-sporing with
peritrichous flagella. Staphylococcus aureus was
The cell toxicity was measured by a MTT method coccus shape, gram positive and grape like
which is a simple non-radioactive colorimetric clusters arrangements were observed.
assay [10]. A549 cells (Lung cancer) and A375
(Melanoma cells) cells were plated out at a Biochemical test: The following observations of
density of 1X104 cells/well in 96-well microtiter biochemical tests were observed for E. coli and
plates. After 24 h incubation, the cells were Staphylococcus aureus.
treated with nano materials (green synthesized
Zinc oxide nanoparticles with Jatropha and 3.1 Antibiotic Susceptibility Test
Copper oxide nanoparticles with Nakara) upto
20ppm for 24 h. followed by incubation, the The Antibiotic susceptibility tests were conducted
media were replaced with 20μl of MTT reagent (5 by placing various HIMEDIA antibiotic disc and
mg/ml) and incubated in 5% CO2 at 37°C for 4 h. antibiotic susceptibility profiles were made by the
DMSO was then added to solubilize the MTT observation of clearance zones formed.

Table 2. Biochemical tests for E. coli and Staphylococcus aureus

Biochemical test Escherichia coli Staphylococcus aureus

Indole test Positive Negative
Voges Proskauer Negative Negative
Citrate utilization test Positive Negative
Catalase test Negative Positive
Coagulase test Negative Positive

1155
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

Table 3. Antibiotic susceptibility test for E. coli and S. aureus

S.no Antibiotics E. coli (mm) Staphylococcus aureus (mm)

1 Ampicillin 4±0.01 9±0.4
2 Cephalosporin 9±0.3 2±0.04
3 Macrolide 12±0.3 5±0.26
4 Monobactam 9±0.2 3±0.09
5 Carbapenem 8±0.2 4±0.08
6 Sulfonamide 7±0.3 7±0.3
7 Nitroimidazole 5±0.25 5±0.2
8 Rifamycin 7±0.4 2±0.03
9 Fluoroquinolone 23±0.9 6±0.1
10 Elfamycin 8±0.3 2±0.05
11 Ceftazidime 5±0.03 2±0.04
12 Cefepime 6±0.04 7±0.2
13 Norfloxacin 3±0.09 5±0.1
14 Levofloxacin 1±0.04 4±0.09
15 Chloramphenicol 6±0.03 8±0.2
16 Tetracycline 2±0.08 9±0.3
17 Streptomycin 9±0.3 1±0.04
18 Augmentin (Amoxicillin & 26±0.6 4±0.03
Potassium Clavulanate)
19 Kanamycin 8±0.2 2±0.07
20 Pencillin – G 2±0.06 6±0.02
21 Gentamycin. 9±0.4 2±0.07
22 Vancomycin 7±0.02 26±0.665
23 Methicillin 9±0.4 8±0.3

The E. coli isolate was sensitive to The antimicrobial activity results of were as
Fluoroquinolone and Augmentin whereas S. shown in the Table 4, Table 5 and Table 6.
aureus was resistant to every antibiotic except
Vancomycin. The green synthesized nano particles with
greater activity were characterized by Scanning
3.2 Molecular Analysis electron microscopy.
The given organisms were identified as E. coli 3.3.1 Toxicity
and S. aureus by 16srRNA analysis. The
phylogenetic analysis was done by ClustalW [9]. The nano materials were found to be non-toxic
and the tree were constructed by Neighbour up to 20ppm concentration on cell lines tested.
joining method. The 16srRNA sequences of E. Green synthesized Zinc oxide nanoparticles with
coli and S. aureus were submitted to NCBI. The Jatropha and Copper oxide nanoparticles with
accession numbers were given below: Nakara’s soluble MTT OD is less than the control
E. coli Accession number: PQ084693 Mitomycin C and Doxorubicin.
S. aureus Accession number: PQ084695
4. DISCUSSION
3.3 Antimicrobial Activity of
Phytochemicals, Nano Particles and Extreme drug resistant bacteria were isolated
Green Synthesized Nano Particles from sewage samples collected in Hyderabad,
Telangana, India. Among 24 antibiotics tested S.
The aqueous and ethanolic extracts of Neem, aureus was found to be sensitive to one
Nakara, Jatropha milk, Mango seed, Clove, antibiotic (vancomycin) whereas E. coli was
Ginger, Cardamom, Cinnamon and Betel were found to be sensitive to two of the antibiotics
tested for antimicrobial activity. The Copper oxide (Fluoroquinolone and Augmentin). Similar
and Zinc oxide nano particles and the green findings of extreme drug resistance bacteria were
synthesized nano particles formed by respective reported in the literature. High resistance to the
phytochemicals were also tested for antimicrobial antibiotics i.e. cephalosporins, fluoroquinolones,
activity. trimethoprim-sulfamethoxazole, and tetracycline

1156
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

by Naziri Z et al. [11]. Nearly all antibiotics highest sensitivity for E. coli and E. faecalis [15].
including frequently using beta lactam The minimum inhibitory concentration of Cu/Zn
combination antibiotics were found resistant [12]. nanomaterials for the E. coli and S. aureus
Out of the 73 isolated strains of S. aureus by strains were of 3.75 and 2.50 mg/ml [16]. 40
Sadat SS et al. [13] 32 were found to be different isolates of subclinical mastitis are
methicillin-resistant S. aureus (MRSA). Among recovered from the milk samples were sensitive
methicillin-resistant S. aureus isolates, 96.8 and to zinc oxide nanoparticles [17]. The study of
12.5% were multi-drug resistance and extreme Abbas ZM et al. [18] showed that the conjugation
drug resistance, respectively. All the methicillin of copper and zinc nanoparticles with the
resistant strains were found to sensitive to classical antibiotics has a great antibacterial
vancomycin [13]. Iqbal Z et al [14] showed high activity 9 Plants aqueous extracts were used
resistant bacteria against cefuroxime, co- for antimicrobial activity and among them
amoxiclav, cefixime, ceftazidime, cefotaxime, Jatropha and Nakara plants aqueous
ceftriaxone, nalidixic acid, ciprofloxacin, extracts were showing more antimicrobial activity
pepedemic acid, norfloxacin, and co-trimoxazole. against gram positive and gram-negative
Nanomaterials particularly Silver (Ag), Copper bacteria. These aqueous extracts were used for
(Cu), Zinc (Zn) were researched as potent green synthesis of copper and zinc oxide
antimicrobials. Copper nanoparticles shown the nanomaterials.

Fig. 1. Neighbour joining tree of E. coli containing 16srRNA gene sequence

Table 4. Antimicrobial activity of Aqueous and ethanolic extracts of phytochemicals collected
from plant samples against E. coli and S. aureus

Sample E. coli (mm) S. aureus (mm)

Aqueous Ethanolic Aqueous Ethanolic
Neem 22 ±0.4 18 ±0.4 20 ±0.6 19 ±0.4
Nakara 23 ±0.3 20 ±0.5 22 ±0.5 19 ±0.3
Jatropha milk 24 ±0.6 19 ±0.4 23 ±0.4 20 ±0.5
Mango seed 15 ±0.2 14 ±0.3 16 ±0.3 12 ±0.1
Clove 14 ±0.1 12 ±0.2 16 ±0.2 15 ±0.1
Ginger 13 ±0.1 11 ±0.1 14 ±0.2 11 ±0.1
Cardamom 17 ±0.3 15 ±0.2 17 ±0.3 16 ±0.2
Cinnamon 15 ±0.2 12 ±0.1 14 ±0.3 11 ±0.2
Betel 16 ±0.2 12 ±0.2 17 ±0.4 15 ±0.3

1157
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

Fig. 2. Neighbour joining tree of S. aureus containing 16srRNA gene sequence

Table 5. Antimicrobial activity of Chemically synthesized nano particles against E. coli

and S. aureus

Nano particles E. coli (mm) S. aureus (mm)

CuO nano particle 19 ±0.4 20 ±0.3
ZnO nano particle 21 ±0.5 20 ±0.4

Table 6. Antimicrobial activity of Green synthesized nano particles against E. coli

and S. aureus

Green synthesised nano E. coli (mm) S. aureus (mm)

particles
ZnO CuO ZnO CuO
synthesized synthesized synthesized synthesized
Neem aq 26 ±0.5 24 ±0.6 26 ±0.7 26 ±0.5
Nakara aq 25 ±0.6 31 ±0.6 25 ±0.6 30 ±0.7
Jatropha milk aq 31 ±0.7 26 ±0.5 30 ±0.7 26 ±0.4
Mango seed aq 18 ±0.4 20 ±0.3 19 ±0.4 21 ±0.3
Clove aq 18 ±0.3 20 ±0.4 19 ±0.2 20 ±0.4
Ginger aq 19 ±0.4 18 ±0.3 18 ±0.3 20 ±0.3
Cardamom aq 18 ±0.4 19 ±0.2 20 ±0.6 20 ±0.4
Cinnamon aq 20 ±0.5 21 ±0.6 19 ±0.4 19 ±0.5
Betel aq 19 ±0.3 20 ±0.4 19 ±0.3 21 ±0.6

The green synthesized nanomaterials sizes were zinc nitrate were of spherical shape with the
with an average of 26nm for ZnO with Jatropa average size of 21.49 and 25.26 [20].
and 25nm for CuO with Nakara. Similar studies Vishveshvar K et al. [21] reported SEM studies of
of Pradheesh G et al. [19] reported Ag2O green synthesized CuO with average size of
nanomaterials of different sizes which are below 300nm.
100nm. The nanomaterials of zinc acetate and

1158
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

Fig. 3. SEM images green synthesized Zinc oxide nanoparticles with Jatropha and Copper
oxide nanoparticles with Nakara

Antimicrobial activities of these green 5. CONCLUSION

synthesized nanomaterials were more when
compared with phytochemical, chemical The present study concludes that the green
synthesized nanomaterials and 30mg synthesized nano particles are effective in
chloramphenicol. Antimicrobial activities were in treating multidrug resistant bacterial infections
accordance to the reports of the nanoparticles over antibiotics and chemically synthesized nano
the inhibition for gram positive and negative particles due to more shelf life, lesser side effects
bacteria with the minimal concentration of and had effective bactericidal properties, in turn
12.5mg/ml. 20±0.7 and 16±0.5 diameter was reducing environmental wastage produced.
observed as the highest inhibition against the S. Green synthesized nano particles retain their
aureus and E. coli strains [22]. Different activity for longer periods than chemically
concentrations of CuSO4 (0.1 and 0.01M) shown synthesized nano particles. The usage of green
antimicrobial effect for the strains of E. coli and synthesized nano particles can be cost effective
S. aureus (33±0.57 and 6±2mm) whereas and efficient compared to other conventional
complete absence of growth is seen in case of S. methods.
aureus in study by Taran M et al. [23]. 98.8 and
99.7 percent efficiency reported against gram DISCLAIMER (ARTIFICIAL INTELLIGENCE)
positive and negative bacteria with CuO/Ag
nanoparticles whereas 91.7 and 89.3 percent Author(s) hereby declare that NO generative AI
efficiency reported with ZnO/Ag nanoparticles technologies such as Large Language Models
against E. coli and S. aureus observed by (ChatGPT, COPILOT, etc) and text-to-image
Asamoah RB et al. [24]. Green synthesis of generators have been used during writing or
nanomaterials is in acceleration due to editing of manuscripts.
ecofriendly process, non-consumption of toxic
chemicals, safer synthesized metals etc.
COMPETING INTERESTS
Nanomaterials are generally regarded as toxic as
Authors have declared that no competing
reported in the studies by Hussain SM et al. [25]
interests exist.
and Sakhtianchi R et al. [26] against mouse
fibroblast cells Hence toxicity studies to
chemically synthesized and green synthesized REFERENCES
nanomaterials were carried out significantly less
toxic when compared with chemical synthesized 1. Spellberg B, Blaser M, Guidos RJ, et al.
nanomaterials. Combating antimicrobial resistance: policy
recommendations to save lives. Clinical
Green synthesized nanomaterials could be Infectious Diseases. 2011;52(Suppl 5):
alternative to antibiotics to control the extreme S397–428.
drug resistance bacteria. DOI:https://doi.org/10.1093/cid/cir153

1159
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

2. Urban-Chmiel R, Marek A, Stępień- 10. Ahmadi M, Hajikhani B, Shamosi A,

Pyśniak D, Wieczorek K, Dec M, Yaslianifard S, Sameni F, Qorbani M,
Nowaczek A, Osek J. Antibiotic resistance Mohammadzadeh M, Dadashi M. Cytotoxic
in bacteria—A review. Antibiotics. 2022; and apoptosis-inducing properties of
11(8):1079. Staphylococcus aureus cytoplasmic extract
DOI:https://doi.org/10.3390/antibiotics1108 on lung cancer cells: Insights from MTT
1079 assay and bax/bcl-2 gene expression
3. Hochma E, Yarmolinsky L, Khalfin B, analysis. Gene Reports. 2024:101955.
Nisnevitch M, Ben-Shabat S, Nakonechny DOI:https://doi.org/10.1016/j.genrep.2024.
F. Antimicrobial effect of phytochemicals 101955
from edible plants. Processes. 2021;9 11. Naziri Z, Derakhshandeh A, Soltani
(11):2089. Borchaloee A, Poormaleknia M,
DOI: https://doi.org/10.3390/pr9112089 Azimzadeh N. Treatment failure in urinary
4. Abdel-Reheem MA, Oraby MM. Anti- tract infections: a warning witness for
microbial, cytotoxicity, and necrotic virulent multi-drug resistant ESBL-
ripostes of Pimpinella anisum essential oil. producing Escherichia coli. Infection and
Annals of Agricultural Sciences. 2015;60 drug resistance. 2020:1839-50.
(2):335-40. DOI: https://doi.org/10.2147/IDR.S256131
DOI:https://doi.org/10.1016/j.aoas.2015.10. 12. Wang M, Wang W, Niu Y, Liu T, Li L, Zhang
001 M, Li Z, Su W, Liu F, Zhang X, Xu H. A
5. Abdelrahman M, Jogaiah S, Abdelrahman clinical extensively-drug resistant (XDR)
M, Jogaiah S. Saponins versus plant Escherichia coli and role of its β-lactamase
fungal pathogens. Bioactive Molecules in genes. Frontiers in microbiology. 2020;11:
Plant Defense: Saponins. 2020:37-45. 590357.
DOI:https://doi.org/10.1007/978-3-030- DOI:https://doi.org/10.3389/fmicb.2020.590
61149-1_4 357
6. Mustapha T, Misni N, Ithnin NR, Daskum 13. Sadat SS, Ahani Azari A. Frequency of
AM, Unyah NZ. A review on plants and multidrug-resistant, extensively drug-
microorganisms mediated synthesis of resistant, and pandrug-resistant
silver nanoparticles, role of plants phenotypes among clinical isolates of
metabolites and applications. International Staphylococcus aureus. Infection
Journal of Environmental Research and Epidemiology and Microbiology. 2020;6(4):
Public Health. 2022;19(2):674. 269-75.
DOI:https://doi.org/10.3390/ijerph1902067 DOI:http://dx.doi.org/10.29252/iem.6.4.269
4 14. Iqbal Z, Mumtaz MZ, Malik A. Extensive
7. Pandit C, Roy A, Ghotekar S, Khusro A, drug-resistance in strains of Escherichia
Islam MN, Emran TB, Lam SE, Khandaker coli and Klebsiella pneumoniae isolated
MU, Bradley DA. Biological agents for from paediatric urinary tract infections.
synthesis of nanoparticles and their Journal of Taibah University Medical
applications. Journal of King Saud Sciences. 2021;16(4):565-74.
University-Science. 2022;34(3):101869. DOI:https://doi.org/10.1016/j.jtumed.2021.0
DOI:https://doi.org/10.1016/j.jksus.2022.10 3.004
1869 15. Ahamed M, Alhadlaq HA, Khan MM,
8. Johnson M, Zaretskaya I, Raytselis Y, Karuppiah P, Al-Dhabi NA. Synthesis,
Merezhuk Y, McGinnis S, Madden TL. characterization, and antimicrobial activity
NCBI BLAST: a better web interface. of copper oxide nanoparticles. Journal of
Nucleic acids research. 2008;36(suppl_2): Nanomaterials. 2014;2014(1):637858.
W5-9. DOI: https://doi.org/10.1155/2014/637858
DOI: https://doi.org/10.1093/nar/gkn201 16. Javadhesari SM, Alipour S,
9. Larkin MA, Blackshields G, Brown NP, Mohammadnejad S, Akbarpour MR.
Chenna R, McGettigan PA, McWilliam H, Antibacterial activity of ultra-small copper
Valentin F, Wallace IM, Wilm A, Lopez R, oxide (II) nanoparticles synthesized by
Thompson JD. Clustal W and Clustal X mechanochemical processing against S.
version 2.0. bioinformatics. 2007;23(21): aureus and E. coli. Materials Science and
2947-8. Engineering: C. 2019;105:110011.
DOI:https://doi.org/10.1093/bioinformatics/ DOI:https://doi.org/10.1016/j.msec.2019.11
btm404 0011

1160
 Nikhil et al.; J. Adv. Biol. Biotechnol., vol. 27, no. 9, pp. 1152-1161, 2024; Article no.JABB.123137

17. Alekish M, Ismail ZB, Albiss B, Nawasrah DOI:https://doi.org/10.1007/s12668-018-

S. In vitro antibacterial effects of zinc oxide 0508-5
nanoparticles on multiple drug-resistant 22. Takele E, Feyisa Bogale R, Shumi G,
strains of Staphylococcus aureus and Kenasa G. Green synthesis,
Escherichia coli: An alternative approach characterization, and antibacterial activity
for antibacterial therapy of mastitis in of CuO/ZnO nanocomposite using Zingiber
sheep. Veterinary world. 2018 ;11(10): officinale Rhizome Extract. Journal of
1428. Chemistry. 2023;2023(1):3481389.
DOI:https://doi.org/10.14202%2Fvetworld. DOI: https://doi.org/10.1155/2023/3481389
2018.1428-1432 23. Taran M, Rad M, Alavi M. Antibacterial
18. Abbas ZM, Mohsin IH, Ahmade N. The activity of copper oxide (CuO)
biological activity of Zinc oxide and copper nanoparticles biosynthesized by Bacillus
oxide nanoparticles against sp. FU4: optimization of experiment
Staphylococcus aurous and Escherichia design. pharmaceutical sciences. 2017;
coli bacteria. Solid State Technology. 2020; 23(3):198-206.
63(6):12957-68. DOI:https://doi.org/10.15171/PS.2017.30
19. Pradheesh G, Suresh S, Suresh J, 24. Asamoah RB, Annan E, Mensah B,
Alexramani V. Antimicrobial and Anticancer Nbelayim P, Apalangya V, Onwona-
Activity Studies on Green Synthesized Agyeman B, Yaya A. A comparative study
Silver Oxide Nanoparticles from the of antibacterial activity of CuO/Ag and
Medicinal Plant Cyathea nilgiriensis ZnO/Ag nanocomposites. Advances in
Holttum. International Journal of Materials Science and Engineering. 2020;
Pharmaceutical Investigation. 2020;10(2). (1):7814324.
DOI: https://doi.org/10.5530/ijpi.2020.2.27 DOI:https://doi.org/10.1155/2020/7814324
20. Fakhari S, Jamzad M, Kabiri Fard H. 25. Hussain SM, Hess KL, Gearhart JM, Geiss
Green synthesis of zinc oxide KT, Schlager JJ. In vitro toxicity of
nanoparticles: a comparison. Green nanoparticles in BRL 3A rat liver cells.
Toxicology in vitro. 2005;19(7):975-83.
chemistry letters and reviews. 2019;12(1):
DOI:https://doi.org/10.1016/j.tiv.2005.06.03
19-24.
4
DOI:https://doi.org/10.1080/17518253.201
26. Sakhtianchi R, Minchin RF, Lee KB,
8.1547925
Alkilany AM, Serpooshan V, Mahmoudi M.
21. Vishveshvar K, Aravind Krishnan MV, Exocytosis of nanoparticles from cells: role
Haribabu K, Vishnuprasad S. Green in cellular retention and toxicity. Advances
synthesis of copper oxide nanoparticles in colloid and interface science. 2013;
using Ixiro coccinea plant leaves and its 201:18-29.
characterization. BioNanoScience. 2018;8: DOI:https://doi.org/10.1016/j.cis.2013.10.0
554-8. 13

Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual
author(s) and contributor(s) and not of the publisher and/or the editor(s). This publisher and/or the editor(s) disclaim responsibility for
any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.
_________________________________________________________________________________
© Copyright (2024): Author(s). The licensee is the journal publisher. This is an Open Access article distributed under the terms
of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.

Peer-review history:
The peer review history for this paper can be accessed here:
https://www.sdiarticle5.com/review-history/123137

1161