A Comparison of Oligodynamic Effects of Copper Nanoparticles and Aluminum

(III) Oxide Nanoparticles against Streptococcus pyogenes

A Introduction of the Research Paper submitted to the Science Department

of the Senior High School Faculty of Makati Science High School

In partial fulfillment of the requirements for the course

Capstone Research

By:

Matthew Rei B. De Leon

Chrisjomar J. Sta. Maria

Rey David John D. Volante

Grade 12 – Josette Biyo

July 2017
 Chapter One

THE PROBLEM AND ITS SETTINGS

Background of the Study

Streptococcus pyogenes, a gram-positive bacteria, is one of the most rampant pathogens

in humans, with 5-15% of normal individuals harbouring the bacterium, usually in the

respiratory tract, without signs of disease. S. pyogenes can infect when its defence mechanism is

compromised or when the organisms are able to penetrate the constitutive defences. When the

bacteria are introduced or transmitted to vulnerable tissues, a variety of infections can occur

(Todar, 2012).

Numerous infections brought by S. pyogenes in humans include scarlet fever, cellulitis,

puerperal sepsis, vulvovaginitis, and even pneumonia. One of the most common infections

brought by this bacterium, especially in kids, is pharyngitis or strep throat (Stevens & Martin,

2014). In a report of the Public Health Agency of Canada (2011), there are 616 million cases of

pharyngitis caused by S. pyogenes worldwide each year. 15-20% of school-aged children has S.

pyogenes in its dormant form in their throats and is more at risk of having the disease.

Furthermore, there are 115.6 million cases of rheumatic heart disease caused by the bacteria

yearly and at least 18.1 million cases of invasive infections, predominantly in older populations

(Public Health Agency of Canada, 2011).

This project aims to compare the oligodynamic effects of copper nanoparticles and

aluminum (III) oxide nanoparticles against Streptococcus pyogenes, a gram positive bacterium.

Results and findings published in numerous scholarly articles suggest the novel ability of both

metal nanoparticles in applying oligodynamic effect against microorganisms.

 In a study by Warnes and Keevil published, it has been proved that copper can prohibit

MRSA spread by touching and fingertip contamination of surfaces. This antimicrobial

application of copper has been projected useful in schools, public places, and especially

hospitals, wherein a main cause of infection is transfer of bacteria through body or fluid contact.

Even application of small amounts of copper in counters, tables, and handrails can drastically

reduce MRSA spread (University of Southampton, 2016). Due to copper’s oligodynamic effect,

the metal can induce a toxic effect in living cells including bacteria. Because bacteria and other

microbes usually house frequently touched surfaces in public places like handrails and

doorknobs, application of copper helps in extinguishing the cells of microorganisms that are on

those surfaces. Other metals and alloys that showcase oligodynamic effect are silver, lead, brass,

and aluminum (Fenlon, 2013).

In another study by Sadiq I.M., et al (2009), metal oxide nanoparticles are known to

possess strong antimicrobial properties. Aluminum (III) oxide, a very common metal oxide used

in industries and households, was tested with its antimicrobial property against the gram-

negative Escherichia coli bacteria. However, mild toxicity effects have been observed in E. coli

in the environment, even in high aluminum (III) oxide concentrations, more likely because of

free-radical scavenging properties of the particles that have prevented cell wall disruption and

drastic antimicrobial action.

With this in mind, the researchers choose to try and compare both metal-containing

nanoparticles to this versatile microorganism and to determine which would have a greater

antibiotic effect on S. pyogenes. This would provide additional knowledge on to preventing the

spread of diseases caused by microorganisms, particularly the array of diseases that is caused by

the bacteria to be studied.

Research Framework

This research project aims to compare the oligodynamic effects of copper nanoparticles

and aluminum (III) oxide nanoparticles against S. pyogenes. This research is guided by the

following framework that will guide the researchers in conducting the project.

Figure 1.1 Research Framework Involving Comparison of Oligodynamic Effects of Copper

Nanoparticles and Aluminum (III) Oxide Nanoparticles against Streptococcus

pyogenes

Several variables are to be considered in conducting the experimentation. The presence of

the two metal-containing nanoparticles is regarded as the independent variable that shall make an

effect to the oligodynamic effect against S. pyogenes. Another important factor to be considered

is the size of the nanoparticles obtained for the experiment in either extraction or purchasing.
Several nanoparticle studies suggest that smaller-sized nanoparticles pose a greater effect since

total surface area is increased when size (diameter in nanometers) of individual nanoparticles is

decreased.

External factors include the entire laboratory conditions as well as the researchers

themselves. To ensure the control of such conditions, aseptic technique is to be strictly followed

in using laboratory apparatus in almost every instant to limit intervention with the

experimentation. The researchers are also to ensure they are in good health condition, and are

regularly sanitized to prevent other microorganisms in contact with the researchers’ bodies to

reach the bacterium cultures under study.

Statement of the Problem

This research project aims to answer the following questions at the end of the study:

MAIN QUESTION: Which of the metal-containing nanoparticles - copper or aluminum (III)

oxide - exhibits a greater oligodynamic effect on Streptococcus pyogenes?

SPECIFIC QUESTIONS:

1. Will the difference of oligodynamic effect of one metal-containing nanoparticle be

statistically

greater than that of the other nanoparticle?

2. Will the oligodynamic effect change if both nanoparticles will be tested against S.

pyogenes?

Research Hypotheses
NULL HYPOTHESIS: Neither of the two metal-containing nanoparticles exhibit a significantly

greater oligodynamic effect on Streptococcus pyogenes.

ALTERNATIVE HYPOTHESIS: Either one of the the two metal-containing nanoparticles

exhibit a significantly greater oligodynamic effect on Streptococcus pyogenes.

Significance of the Study

This study aims to compare the oligodynamic effects of copper nanoparticles and

aluminum (III) oxide nanoparticles against Streptococcus pyogenes, a versatile species of

bacteria responsible for a wide variety of diseases and complications. Since both metal-

containing nanoparticles are proven to be antimicrobial through past researches, this study serves

as an extension.

Furthermore, the knowledge that will be derived from this study will serve as basis to

benefit facilities which are potential breeding grounds for bacteria, specifically S. pyogenes, like

hospitals, clinics, and other potential microbial environments. Being able to determine the

comparison between nanoparticles will provide more solutions onto how will the spread of

bacteria be limited. This will benefit the general public, as well as medical professionals, which

are in constant contact with potential breeding grounds for bacteria (i.e. hand railing,

countertops, and other furnitures).

Scope and Limitation of the Study

For this study only two metal-containing nanoparticles will be concerned in this study:

copper and aluminum (III) oxide nanoparticles. Likewise, only the S. pyogenes will serve as

subject microorganisms in this study.

 Furthermore, this study is only concerned on the antimicrobial effect of these

nanoparticles against the bacterium species. The research will not tackle the process on how the

nanoparticles disrupt the proliferation of the bacteria. Copper and aluminum (III) oxide

nanoparticles will be compared in this study; the two will also be tested together to determine if

there is a change if both are used at the same bacterium culture. Zones of inhibition will be

determined to compute for the antimicrobial indices, which will be the data to be used for

statistical treatment.

Definition of Terms

Antimicrobial- acts an agent to inhibit the growth of microorganisms, specifically, the

pathogenic organisms (Merriam Webster, n.d.) .

Free radicals- are atoms or groups of atoms with an odd (unpaired) number of electrons and can

be formed when oxygen interacts with certain molecules.

Gram- negative bacteria - are microorganisms that do not retain crystal violet dye in gram

staining protocol. Their cell envelope contains an additional outer membrane composed by

phospholipids and lipopolysaccharides which face the external environment.

Gram- positive bacteria - are microorganisms that retain the crystal violet in gram staining

protocol.

Metal Oxides- are compounds formed by combining metal and oxygen. They are usually solids

at room temperature and if dissolved, it forms alkaline solutions.

Nanoparticles- are microscopic particles whose dimensions are less than 100 nm (ScienceDaily,

n.d.).
Oligodynamic Effect- result of metal ions in metallic substances having a toxic and antibiotic

effect on molds, spores, viruses, and other living cells such as bacteria (Ly, 2014).

Transmission Electron Microscope- a form of electron microscope in which an image is derived

from electrons that have passed through the specimen, in particular one in which the whole

image is formed at once rather than by scanning.

 Chapter Two

REVIEW OF RELATED LITERATURE

Oligodynamic action is the ability of small amounts of heavy metals to exert a lethal

effect on bacterial cells.Oligodynamic metals, such as silver and copper, have long been utilized

as disinfectants for non-spore-forming bacteria and viruses (Thurman & Gerba 1988). Many

metallic elements have been observed to inhibit the growth of bacteria and to inactivate enzymes.

Practical application of such activity of metals has been made in the purification of water and in

the preservation of tomato juice, cider and hides. This antimicrobial effect is shown by metals

such as mercury, silver, copper, lead, zinc, gold, aluminum and other metals, and the

concentration of the metal needed for this antimicrobial effect is extremely small (Jain 1990).

In a study conducted by Shreshta, Joshi, Gopali, and Piya in 2009, they aim to evaluate

the effect of oligodynamic metals such as copper, silver and brass against enteric gram negative

drinking water isolates such as Salmonella paratyphi, Shigella spp., E. coli (MDR), E. coli,

Vibrio cholerae and Klebsiella spp. in Kathmandu. The test was carried out by preparing broth of

the respective microorganisms followed with contaminating autoclaved distilled water with 1%

(by volume) of the prepared broth culture and incubating the contaminated water in the

respective metal pots up to 48 h (holding time). Reduction in the microbial load was assessed by

pour plating the water content in the metal pots on Nutrient agar medium in every 0, 4th, 8th,

12th, 24th and 48th h of incubation (holding time). Among three test pots, copper pot showed the

maximum bactericidal action compared to silver and brass pots towards most of the enteric gram

negative bacterial isolates of water. Complete inhibition of tested organism was recorded within

4 to 48 hours of holding time. This study suggested the promotion of use of water pots made of
oligodynamic metals such as silver and copper, and alloy such as brass to control the gram

negative enteric pathogens in drinking water.

The high occurrence of resistance of different microorganisms to the majority of

antimicrobial agents is attracting a great deal of attention. The World Organization for Animal

Health, Food and Agriculture Organization, and World Health Organization have all commented

on the serious threat posed by antimicrobial-resistant pathogenic organisms to human and animal

health (Raffi, et.al, 2010). Resistance of microorganisms to antibiotics is steadily rising, with

reports showing that quite a number of the recognized antimicrobial agents in existence have

demonstrated resistance by one species of microorganism or another, so basically there is no

single antimicrobial agent available for human and animal use that has not demonstrated

resistance by microorganisms. This development had compelled clinicians to rely on in vitro

antimicrobial susceptibility testing for diagnostic purposes. In this regard, synthesis or extraction

of compounds such as nanoparticles with antimicrobial properties is essential, and has potentially

promising applications in the fight against the ever-growing number of antimicrobial-resistant

pathogenic microorganisms which pose a continuous threat to human and animal health.

Nanotechnology is a field of science with vast potential in medicine. Being analogous

with nature, the combination of nanoscience and biology will not only strengthen the fight

against pathogenic microorganisms but can also result in a change in approach towards

combating infectious diseases. Consequently, diseases like cancer and rheumatoid arthritis are

also being combated using nanoparticles. Materials in the range of 100 nm or less are considered

to be nanoparticles. They exhibit a wide range of properties, including optical, electrical,

catalytic, magnetic, and biological activity, which is different from that of their bulk materials.

Some of the biological properties of nanoparticles have been explored by antimicrobial

susceptibility testing of nanoparticles produced from different metals using different synthetic

methods. It has been reported that metal nanoparticles (Ag, Cu, CuO, Au) exhibit a wide

spectrum of antimicrobial activity against different species of microorganisms, including fungi

and Gram-positive and Gram-negative bacteria. Antibacterial activity has been reported against

Escherichia coli and a nonresistant strain of Gram-positive bacteria (Staphylococcus aureus),

and the results obtained indicate inhibition of growth of the organism due to addition of the

nanoparticles.

In the research entitled “Synthesis, characterization, and antimicrobial properties of

copper nanoparticles” by Usman, El Zowalaty, Shameli, Zainudinn, Salama, and Ibrahim, the

scientists prepared pure copper nanoparticles in the presence of a chitosan stabilizer through

chemical means. The purity of the nanoparticles was authenticated using different

characterization techniques, including ultraviolet visible spectroscopy, transmission electron

microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and field emission

scanning electron microscopy. The antibacterial as well as antifungal activity of the

nanoparticles were investigated using several microorganisms of interest, including methicillin-

resistant Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Salmonella

choleraesuis, and Candida albicans. The effect of a chitosan medium on growth of the

microorganism was studied, and this was found to influence growth rate. The size of the copper

nanoparticles obtained was in the range of 2–350 nm, depending on the concentration of the

chitosan stabilizer.

Zerovalent copper nanoparticles (Cu0) of 12 nm size were synthesized using an inert gas

condensation method in which bulk copper metal was evaporated into an inert environment of

argon with subsequent cooling for nucleation and growth of nanoparticles. Crystalline structure,
morphology and estimation of size of nanoparticles were carried out by X-ray diffraction and

transmission electron microscopy. The antibacterial activity of these nanoparticles against the

Gram-negative bacterium Escherichia coli was assessed in liquid as well as solid growth media.

It was observed from scanning electron microscopic analysis that the interaction of copper

nanoparticles with E. coli resulted in the formation of cavities/pits in the bacterial cell wall. The

antibacterial property of copper nanoparticles was attributed mainly to adhesion with bacteria

because of their opposite electrical charges, resulting in a reduction reaction at the bacterial cell

wall. Nanoparticles with a larger surface-to-volume ratio provide more efficient means for

antibacterial activity (Raffi, et.al, 2010).

The bactericidal activity of catechins has been reported to be lower against Gram-

negative bacteria compared with that against Gram-positive bacteria ( Ikigai et al. 1993 ), and

this has been attributed to the presence of a strong negative charge on the lipopolysaccharide cell

surface of Gram-negative bacteria (Ikigai et al. 1993 ).

Recently, the bactericidal activity of catechin-Cu(II) complexes against Escherichia coli,

a Gram-negative bacterium ( Kimura et al. 1998 ; Hoshino et al. 1999 ) was reported, and it was

shown that the presence of a non-lethal concentration of Cu(II) dramatically increased the

bactericidal activity of catechins. A mechanism was proposed in which Cu(II) was reduced to

Cu(I) by catechins, and reoxidation of Cu(I) to Cu(II) by molecular oxygen generated hydrogen

peroxide on the cell surface, which was responsible for the bactericidal activity of the catechin-

Cu(II) complex ( Hoshino et al. 1999 ). At physiological pH, catechins are uncharged, but in the

presence of Cu(II), catechins are complexed ( Cetta et al. 1977 ; Weber 1988) and the resultant

catechin-Cu(II) complexes are considered to be positively charged. Therefore, the negative

charge on the cell surface of Gram-negative bacteria may attract catechin-Cu(II) complexes more
than catechin alone. Formation of catechin-Cu(II) complexes resulted in an increase in the

bactericidal activity of catechin against Gram-negative bacteria.

In this work of Hoshino et.al., the bactericidal activity of catechin-copper (II) complexes

against Staphylococcus aureus compared with Escherichia coli was investigated in relation to the

generation of hydrogen peroxide and the binding of Cu(II) ion onto the bacteria. The bactericidal

activity of catechin-Cu(II) complexes against Staph. aureus (Gram-positive) was much lower

than that against E. coli (Gram-negative), suggesting that the binding of copper ions to the

surface of bacterial cells plays an important role in the bactericidal activity of catechin-Cu(II)

complexes.

The treatment and cleaning of medical instruments and devices, water purification, food

industry are closely connected with the development of effective, environmentally friendly,

available and low cost bactericide compositions for pathogenic microorganism control. Due to a

wide presence in nature, implementation of different functions within the majority of living

organisms, relatively low cost and environmental safety, copper compounds (Cu) have a high

potential for their application as antibacterial agents being capable to replace silver and

composites of different precious metals.

Thus, it has been demonstrated that Cu nanosized particles are able to show antimicrobial

properties in relation to a wide range of microorganisms including pathogenic bacteria. On the

one hand, according to, Cu nanoparticles are hypotoxic. On the other hand, they show a high

antibacterial effect in relation to the cells of test cultures of G+ and G- microbacteria, which

makes reasonable their application as part of wound-healing preparations. At the same time, Cu

nanoparticles are demonstrated to have high efficiency for application in bactericide plasters or

bandages due to high antimicrobial activity to pathogenic microorganisms and the illegible
sensibility of human tissues to copper compounds. Other authors have shown that Cu

nanoparticles may significantly increase the antibacterial activity of sorbents exposed to

Escherichia coli cultures owing to the copper capability to disrupt metabolism of a microbial cell

while interacting with microorganisms. In other words, the production of bactericidal plasters,

bandages and medicines with the inclusion of copper nanoparticles and copper ions may have a

great potential in terms of their biomedical application. However, the lack of experimental data

in the literature on antibacterial properties linked to physicochemical parameters of nanoparticles

in water suspensions prevents forecasting their antibacterial activity and, respectively, preparing

widely used antibacterial suspensions.

In this study by Godymchuk, Frolov, Gusev, Zakharova, Yunda, Kuznetsov, and

Kolesnikov, the researchers points out that the production of bactericidal plasters, bandages and

medicines with the inclusion of copper nanoparticles and copper ions may have a great potential

in terms of their biomedical application. The work considers the influence of the synthesis

conditions, size, aggregation status, and charge of nanoparticles in aqueous solutions as well as

the type of microorganisms to the antibacterial properties of water suspensions of

electroexplosive copper nanoparticles in the conditions in vitro in relation to strains Escherichia

coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus cereus. Water dispersions

of copper nanoparticles were shown to inhibit the growth of test cells for both G+ and G-

microbacteria but the degree of such an influence strongly depended on the type of a test strain.

The authors have demonstrated that use of deeply purified water and alcohol-containing

stabilizers at the synthesis of nanoparticles via metals electric erosion in the liquid prevents the

copper nanoparticles coagulation and significantly influences on their physicochemical

characteristics and, consequently, antibacterial properties.

 The pandemic of hospital acquired infections caused by methicillin-resistant

Staphylococcus aureus (MRSA) has declined but the evolution of strains with enhanced

virulence, toxins and the increase of community-associated infections is still a threat. In previous

studies, simulated droplet contamination of MRSA was killed on copper and brass surfaces

within 90 minutes. However, contamination of surfaces is often via finger tips which dries

rapidly and may be overlooked by cleaning regimes unlike visible droplets. In this new study a

5-log reduction of a hardy epidemic strain of MRSA (EMRSA-16) was observed following 10

minutes contact with copper and 4-log reduction observed on copper nickel and cartridge brass

alloys in 15 minutes. A methicillin-sensitive strain (MSSA), from an osteomyelitis patient, was

killed on copper surfaces in 15 minutes and a 4-log and 3-log reduction occurred within 20

minutes contact with copper nickel and cartridge brass, respectively. Bacterial respiration was

compromised on copper surfaces and superoxide generated as part of the killing mechanism. In

addition, destruction of genomic DNA occurs on copper and brass surfaces allaying concerns

about horizontal gene transfer and copper resistance. Incorporation of copper alloy biocidal

surfaces could help to reduce the spread of this dangerous pathogen (Warnes and Keevil, 2016).

When the dimensions of a material are reduced to the atomic level it attains a range of

unique properties that can be manipulated suitably for the desired applications. Because most of

the biological processes also take place on the nanoscale, synergistic application of

nanotechnology and biology can possibly address several important biomedical

problems.Currently nanosized organic and inorganic nanoparticles (NPs) are finding increasing

applications in medical devices as a result of their amenability to biological functionalization.

Antimicrobial agents are highly relevant for a host of industrial applications in

environmental, food, synthetic textiles, packaging, healthcare, and medical care products. They
can be grouped broadly into two types: organic and inorganic. The inorganic materials have

gained importance over the past decade because of their ability to withstand adverse processing

conditions. The antibacterial activity is known to be a function of the surface area in contact with

the microorganisms; a larger surface area (as in the case of NPs) ensures a broad range of

probable reactions with bioorganics present on the cell surface, as well as environmental

inorganic and organic species.

Yamamoto et al investigated cytotoxic effects of metal oxide particles (Al2O3, titanium

dioxide [TiO2], and zirconium oxide [ZrO2]) on murine fibroblasts and murine monocyte

macrophages. They studied cytotoxicity as a function of shape, size, and surface area of the

particles and also compared toxicity behavior of bulk and nanomaterials. Hanawa et al studied

the toxicity of a range of metal oxide NPs, including Al2O3, having 500-nm to 3000-nm

diameter on human fibroblast cells. Several recent studies with alumina NPs showed

considerable cytotoxic effect on the mammalian cells.There have been very few studies available

in the literature on the interaction of the alumina NPs with microbes. One past study found no

detrimental effect of an alumina slurry between 62.5 and 250 mg/L concentration range on E.

coli. Considering the paucity of prior literature reports on the effect of alumina NPs on bacterial

species, the objective of the present work was to investigate the possible growth-inhibitory effect

of alumina NPs over a wide concentration range (10–1000 μg/mL) on an environmentally

relevant gram-negative model microorganism, E. coli. The implications of this type of growth-

inhibitory study are mainly twofold: A. For biomedical applications the bactericidal properties of

ceramic oxides must be studied. B. The growth inhibition effects of the oxides may reflect on

toxicity behavior of metal oxides on bacterial species in the environment.

 Metal oxide nanoparticles (NPs) are known to possess strong antimicrobial properties.

Aluminum oxide NPs have wide-range applications in industrial as well as personal care

products. In the absence of prior reports on the antimicrobial properties of alumina NPs for a

wide concentration range, the principal objective of the present work was to study the growth-

inhibitory effect of alumina NPs over a wide concentration range (10–1000 μg/mL) on an

environmentally relevant gram-negative model microorganism, Escherichia coli. The mean

diameter of the NPs was determined to be 179 nm in aqueous dispersion used for this study, and

surface area was determined to be 21.23 m2 /g. The concentration of 1000 μg/mL was found to

be moderately inhibitory for bacteria. Almost negligible dependence of growth rate on the

concentration of the NPs was observed. The extracellular protein content was found to be

slightly lower in case of cells interacting with 1000 μg/mL alumina than the uninteracted control

cells. Fourier transform–infrared studies established differences in structure between interacted

and uninteracted cells. Alumina NPs showed a mild growth-inhibitory effect, only at very high

concentrations, which might be due to surface charge interactions between the particles and cells.

Free-radical scavenging properties of the particles might have prevented cell wall disruption and

drastic antimicrobial action. This laboratory-scale study suggests that alumina NPs may only

exhibit mild toxicity toward microorganisms in the environment (Sadiq et al, 2009).

The emerging infectious diseases and the development of drug resistance in the

pathogenic bacteria and fungi at an alarming rate is a matter of serious concern. Despite the

increased knowledge of microbial pathogenesis and application of modern therapeutics, the

morbidity and mortality associated with the microbial infections still remains high. Therefore,

there is a pressing demand to discover novel strategies and identify new antimicrobial agents

from natural and inorganic substances to develop the next generation of drugs or agents to
control microbial infections. Prior to the extensive use of chemotherapeutics in modern health

care system, inorganic antimicrobials such as silver and copper were used since ancient times to

treat microbial infections. In the recent times, the advances in the field of nanosciences and

nanotechnology has brought to fore the nanosized inorganic and organic particles which are

finding increasing applications as amendments in industrial, medicine and therapeutics, synthetic

textiles and food packaging products. Nanoparticles usually ranging in dimension from 1-100

nanometers (nm) have properties unique from their bulk equivalent. With the decrease in the

dimensions of the materials to the atomic level, their properties change. The nanoparticles

possess unique physico-chemical, optical and biological properties which can be manipulated

suitably for desired applications. Moreover, as the biological processes also occur at the

nanoscale and due to their amenability to biological functionalization, the nanoparticles are

finding important applications in the field of medicine. The nanoparticles are broadly grouped

into organic and inorganic nanoparticles. The latter have gained significant importance due to

their ability to withstand adverse processing conditions. Currently, the metallic nanoparticles are

thoroughly being explored and extensively investigated as potential antimicrobials. The

antimicrobial activity of the nanoparticles is known to be a function of the surface area in contact

with the microorganisms. The small size and the high surface to volume ratio i.e., large surface

area of the nanoparticles enhances their interaction with the microbes to carry out a broad range

of probable antimicrobial activities. Metal nanoparticles with antimicrobial activity when

embedded and coated on to surfaces can find immense applications in water treatment, synthetic

textiles, biomedical and surgical devices, food processing and packaging. Moreover, the

composites prepared using metal nanoparticles and polymers can find better utilization due to the

enhanced antimicrobial activity.

 This study by Ravishankar Rai and Jamuna Bai in 2011 states that emerging infectious

diseases and the increase in incidence of drug resistance among pathogenic bacteria have made

the search for new antimicrobials inevitable. In the current situation, one of the most promising

and novel therapeutic agents are the nanoparticles. The unique physiochemical properties of the

nanoparticles combined with the growth inhibitory capacity against microbes has led to the

upsurge in the research on nanoparticles and their potential application as antimicrobials. From

centuries metals such as silver have been used for treating burns and chronic wounds, and copper

has been used to make water potable. It is quite evident that some of the metallic compounds

possess antimicrobial property. Recently, the confluence of nanotechnology and biology has

brought to fore metals in the form of nanoparticles as potential antimicrobial agents.

Nanoparticles have unique and well defined physical and chemical properties which can be

manipulated suitably for desired applications. Moreover, their potent antimicrobial efficacy due

to the large surface area to volume ratio has provided them an edge over their chemical

counterparts which are facing the problems of drug resistance. This review focuses on the

properties of different types of metallic nanoparticles such as copper, aluminium, gold, silver,

magnesium, zinc and titanium nanoparticles. The mechanism of action of nanoparticles as

bactericidal, antifungal and antiviral agents will be highlighted in this study. The potential

application of nanoparticles will be also reviewed. The application of nanoparticles as

antimicrobials is gaining relevance in prophylaxis and therapeutics, in medical devices, food

industry and textile fabrics. The problems related to toxicity of nanoparticles will be addressed in

brief.

The work of Lara, Ayala-Nuñez, Ixtepan Turrent, and Rodriguez Padilla narrates that

infections caused by drug-resistant microorganisms result in significant increases in mortality,

morbidity, and cost related to prolonged treatments. The antibacterial activity of silver

nanoparticles against some drug-resistant bacteria has been established, but further investigation

is needed to determine whether these particles could be an option for the treatment and

prevention of drug-resistant microbial infections. Hence, we challenged different drug-resistant

pathogens of clinical importance (multidrug-resistant Pseudomonas aeruginosa, ampicillin-

resistant Escherichia coli O157:H7 and erythromycin-resistant Streptococcus pyogenes) with a

suspension of silver nanoparticles. By means of a luciferase-based assay, it was determined that

silver nanoparticles inactivate a panel of drug-resistant and drug-susceptible bacteria (Gram

positive and Gram negative), exert their antibacterial activity through a bactericidal rather than

bacteriostatic mechanism, and inhibit the bacterial growth rate from the time of first contact

between the bacteria and the nanoparticles. Additionally, strains with a resistant phenotype to

silver nanoparticle were developed and used to explore the bactericidal mode of action of silver

nanoparticles. Through a Kirby–Bauer test, it was shown that silver nanoparticles’ general

mechanism of bactericidal action is based on inhibition of cell wall synthesis, protein synthesis

mediated by the 30s ribosomal subunit, and nucleic acid synthesis. Our data suggest that silver

nanoparticles are effective broad-spectrum biocides against a variety of drug-resistant bacteria,

which makes them a potential candidate for use in pharmaceutical products and medical devices

that may help to prevent the transmission of drug-resistant pathogens in different clinical

environments.

Titania materials are widely used in various research fields due to their several unique

features. For example, titania beads have been known as effective adsorbents specific for

phosphorylated peptides, and nanocrystalline TiO2 electrodes have been applied in the research

of solar cells. The ability of titania in photocatalytic reduction is applied in metal reduction to
remove heavy metals from wastewater. Furthermore, titania materials also have antimicrobial

activities. Previous studies have demonstrated that titania materials can inhibit the cell growth of

microorganisms via photochemical reactions. However, these materials have no selectivity for

specific microorganisms. Additionally, gold nanoparticles and magnetic nanoparticles have been

used as either antibiotic agents or bacterial capture probes. We have demonstrated that

immunoglobulin G (IgG)-bound magnetic nanoparticles can recognize several pathogenic

bacterial strains, including Staphylococcus aureus, Staphylococcus saprophyticus, and

Streptococcus pyogenes, using pseudo-immune interactions. The recognition by nanoprobes of

pathogenic bacteria is based on pseudo-immune interactions between the Fc sites of IgG

molecules and the binding proteins on the surfaces of these bacteria. IgG-bound magnetic

nanoparticles have broadband affinity for pathogenic bacteria, which have binding affinities with

the Fc site of IgG molecules.

Chen,W., Tsai, and Chen, Y. (2008) believe that photokilling approach for pathogenic

bacteria is demonstrated using a new type of magnetic nanoprobe as the photokilling agent. In

addition to their magnetic property, the nanoprobes have other features including a

photocatalytic property and the capacity to target bacteria. The nanoprobes comprise iron

oxide/titania (Fe3O4@TiO2) core/shell magnetic nanoparticles. As dopamine molecules can

self-assemble onto the surface of the titania substrate, dopamine is used as the linker to

immobilize succinic anhydride onto the surfaces of the Fe3O4@TiO2 nanoparticles. This is

followed by the immobilization of IgG via amide bonding. We demonstrate that the IgG–

Fe3O4@TiO2 magnetic nanoparticles not only have the capacity to target several pathogenic

bacteria, but they also can effectively inhibit the cell growth of the bacteria targeted by the

nanoparticles under irradiation of a low-power UV lamp within a short period. Staphylococcus

saprophyticus, Streptococcus pyogenes, and antibiotic-resistant bacterial strains, such as

multiantibiotic-resistant S. pyogenes and methicillin-resistant Staphylococcus aureus (MRSA),

are used to demonstrate the feasibility of this approach.

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