ANTIBACTERIAL POTENTIAL OF BIOLOGICALLY

SYNTHESIZED MAGNESIUM OXIDE

NANOPARTICLES USING SACCHAROMYCES
CEREVISIAE AGAINST GRAM-NEGATIVE BACTERIA
BY
AQSA SHAHID
ROLL NO: 1422
REGISTRATION NO: 2015-GCUF-18531
UNDER THE SUPERVISION OF
DR. MOHSIN KHURSHID

Department of Microbiology
Government College University, Faisalabad
Introduction

 Antibiotics the most important medical innovations in human history. The introduction of a
broad range of antibiotics drastically reduced the morbidity and mortality from infectious
diseases over the last century
 However, the emergence of antimicrobial resistance, which has been widely documented to
worsen clinical outcomes and increase the economic burden of infectious diseases
 Multidrug-resistant (MDR) bacteria are becoming a growing public health crisis and make
many healthcare-associated infections difficult to treat with current antibiotics
Nanoparticles as alternatives

 The application of nanoparticles (NPs) provides a potential strategy to manage infections

caused by these MDR bacterial pathogens
 NPs have shown therapeutic promise owing to their unique physiochemical properties.
 NPs exhibiting antibacterial activities can target multiple biomolecules and have the
potential to reduce or eliminate the evolution of MDR bacteria.

(Lee et al 2019)
 Nanoparticles

 Several characteristics of NPs make them

alternatives to traditional antibiotics.
• The large surface-area-to-volume ratio of
NPs increases the contact area with target
organisms.
• The NPs may enhance the inhibitory effects
of antibiotics.
• The combinations of antibiotics and NPs
provide complex antimicrobial mechanisms
to overcome antibiotic resistance
(Wang et al 2017)
 Magnesium Oxide (MgO) Nanoparticles

 Magnesium oxide (MgO) nanoparticles have attracted interest for use in biomedical

applications due to their antimicrobial properties
 The few studies have reported the antibacterial effect of chemically synthesized magnesium
oxide nanoparticles against some microorganisms.
 Even though the previous studies did prove that MgO nanoparticles had antibacterial
properties under different conditions.

(Nguyen et al 2018)
 Biologically synthesized MgO NPs

 Biologically synthesized NPs are far superior, in several ways, to the chemically synthesized NPs.
 Although the chemical synthesis can produce large quantities of nanoparticles with a defined size
and shape in a relatively short time, they are complicated, costly, and inefficient and produce
hazardous toxic wastes that are harmful, not only to the environment but also to human health.
 There are several reports on the preparation of MgO NPs using the plants and synthesis of MgO NP
from bacterial and fungal sources is missing in literature

(Iravani et al 2016)
Objectives

 Biological synthesis of MgO nanoparticles using yeast Saccharomyces cerevisiae

 Characterization of MgO nanoparticles by several techniques

 Bactericidal potential of MgO nanoparticles against Gram-negative bacteria

Materials and Methods
Clinical isolates

 Clinical isolates were obtained from the Department of Microbiology, Government College
University Faisalabad
 To check the morphological characteristics and confirmation of these isolates API20E and
several biochemical tests were performed
 Catalase
 Oxidase
 Citrate
 Urease
 Triple sugar iron agar
Characterization of nanoparticles

For the characterization of biologically synthesized MgO NPs, various analytical tools were
used
 UV-Visible spectroscopy
 Scanning electron microscopy (SEM)
 Fourier transform infrared (FTIR) spectroscopy
Antibacterial susceptibility testing

 To check the antimicrobial susceptibility profile of isolates, the standard Kirby disc diffusion
method for a list of antimicrobial agents followed by determination of MIC for ciprofloxacin
was used.
Antibacterial activities of MgO nanoparticles

Antibacterial activities were checked using various methods including

 Agar well diffusion method
 Minimum inhibitory concentration (MIC) of MgO NPs
 Minimum bacterial concentration (MBC) of MgO NPs
 Checkerboard assay
 Time-Kill Assay
Antioxidant potential of MgO NPs

 To determine the scavenging activity (free radical) of biologically synthesized magnesium

oxide nanoparticles stable radical i.e., 2,2- diphenyl-1-picrylhydrazyl (DPPH) was used.
Results

UV-Vis spectroscopy
Ultraviolet–visible spectroscopy
0.600

0.500

0.400
Ansorbance

0.300

0.200

0.100

0.000
150.0 200.0 250.0 300.0 350.0 400.0 450.0 500.0 550.0 600.0 650.0

Wavelength (nm)
Scanning electron microscopy
Fourier transform infrared spectroscopy
Antimicrobial susceptibility profile for Escherichia coli and
Klebsiella Pneumoniae

Antimicrobial Agents
Bacterial
species
AMP PRL AMC ATM SAM TZP CRO CTX CAZ FEP IPM MEM AK CN TOB CIP LEV SXT DO

E. coli R R R R R R R R R R R R R R R R R R S

K. Pneumoniae R R R R R R R R R R R R R R S R R R R
 Antimicrobial susceptibility profile of Acinetobacter baumannii and
Pseudomonas aeruginosa strains

Antimicrobial Agents
Bacterial species
AMP PRL AMC ATM SAM TZP CRO CTX CAZ FEP IPM MEM AK CN TOB CIP LEV SXT DO

Acinetobacter
R R R R R R R R R R R S R S R R R R
baumannii

Pseudomonas
R R R R R R NT NT R R S S R R R R R NT NT
aeruginosa
 Antimicrobial susceptibility profile of Salmonella enterica and Proteus
mirabilis strains

Antimicrobial Agents

Bacterial species
AZM
AMP PRL AMC TZP CRO CTX FEP IPM MEM AK CN TOB CIP LEV SXT DO

S
Salmonella enterica serovar Typhi R NT R R R R R S S NT NT NT R R R R

NT
Proteus mirabilis R R R R NT NT R R R S S S R R R R
MIC and MBC for the clinical isolates (Ciprofloxacin)

Bacterial isolates Breakpoints (µg/ml) MIC (µg/ml) MBC (µg/ml)

A. baumannii 4 32 128

P. aeruginosa 2 8 16

E. coli 1 16 64

K. pneumoniae 1 32 64

P. mirabilis 1 8 32

S. Typhi 1 8 16
Zone of Inhibition of NPs against bacterial strains by Agar well diffusion method

Zone of inhibition at different concentration

Bacterial strains
6.25 μg/mL 12.5 μg/mL 25 μg/mL 50μg/mL
Pseudomonas
Not detected Not detected 14 mm 17.5 mm
aeruginosa
Acinetobacter
Not detected Not detected 16 mm 27 mm
baumannii
Escherichia coli Not detected Not detected 13 mm 20 mm
Proteus mirabilis Not detected Not detected 11.5 mm 19 mm
Salmonella Typhi Not detected Not detected 9 mm 15.5 mm
Klebsiella pneumoniae Not detected Not detected 10 mm 21 mm
Synergy of NPs with antibiotic

MIC (µg/ml)
Bacteria Agents FIC FICI Interpretation
Alone Combination
CIP 32 16 0.5
A. baumannii 0.75 Partial synergistic
NP 50 12.5 0.25
CIP 8 4 0.5
P. aeruginosa 0.75 Partial synergistic
NP 100 25 0.25
CIP 16 4 0.25
E. coli 0.5 Synergistic
NP 50 12.5 0.25
CIP 32 8 0.25
K. pneumoniae 0.75 Partial synergistic
NP 50 25 0.5
CIP 8 2 0.25
P. mirabilis 0.5 Synergistic
NP 100 25 0.25
CIP 16 4 0.25
S. Typhi 0.5 Synergistic
NP 100 25 0.25
Time kill Assay
Time kill Assay
Time kill Assay
Antioxidant activity

1.4

1.2

0.8
Absorbance

NP (50 μg/mL )
0.6

0.4

0.2

0
505.0 510.0 515.0 520.0 525.0 530.0 535.0 540.0 545.0 550.0

Wavelength (nm)
Conclusions

 The results have shown that MgO were effective against P. aeruginosa, S. enterica, and P. mirabilis
with 100 μg/ml whereas and for E. coli, K. pneumoniae, and A. baumannii at 50 μg/ml
concentrations.
 The MBC for P. aeruginosa, S. enterica, and P. mirabilis were 200 μg/ml, and 100 μg/ml for E. coli,
K. pneumoniae, and A. baumannii.
 The synergetic effect of ciprofloxacin and MgO NPs was observed for E. coli, P. mirabilis and S.
Typhi whereas a partial synergistic effect for A. baumannii, P. aeruginosa and K. pneumoniae.
 MgO NPs also possess good antioxidants activities
 The MgO NPs can used as potential therapeutic agents against the infections caused by MDR
bacterial pathogens
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 Nguyen NY, Grelling N, Wetteland CL, Rosario R, Liu H. Antimicrobial activities and mechanisms
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