Volume 24, Issue 3 (Aug - Sep 2020)                   2020, 24(3): 234-245 | Back to browse issues page


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Naddafi S, Partoazar A, Dargahi Z, Soltan Dallal M M. Antibacterial Effect of Zinc Oxide Nanoparticles on Standard Strains and Isolates of Pseudomonas Aeruginosa and Staphylococcus Aureus. Journal of Inflammatory Diseases. 2020; 24 (3) :234-245
URL: http://journal.qums.ac.ir/article-1-2899-en.html
1- Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
2- Experimental Medicine Research Center, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
3- Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. , msoltandallal@gmail.com
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1. Introduction

Pseudomonads are non-fermenting bacteria that are widely distributed in the environment. Pseudomonas aeruginosa (P. aeruginosa) is an important opportunistic pathogen that can quickly become resistant to drugs. It causes lung infections in people with cystic fibrosis and those need artificial respiration [1].
Staphylococci are among the first known human pathogens that colonize the skin and mucous membranes [3]. One of the most important species of this pathogen is Staphylococcus aureus (S. aureus) which has become one of the major public health concerns due to its inherent ability and resistance to antimicrobial agents and drugs [4].
In recent years, metal oxides such as zinc oxide (ZnO), have been considered as antimicrobial compounds. ZnO nanoparticles inhibit bacterial growth by producing hydrogen peroxide and penetrating the cell wall and destroying the membrane; but its mechanism of action is still unclear [7-9].
In the study by Azam et al., the antibacterial effect of several nanoparticles on gram-positive and gram-negative bacteria was investigated. Their results showed that nanoparticles of ZnO had a better effect on both groups of bacteria [11]. In a study by Liu et al., ZnO nanoparticles could potentially be used as an effective antibacterial agent to protect the agriculture and food safety [13]. Due to a daily need for food in humans, any change in the food quality and quantity can have a significant impact on the community health. Removal of microbial contamination from food is important at any stages of food production, storage and supply [16]. The aim of this study was to compare the antibacterial effect of ZnO nanoparticles on standard strains of P. aeruginosa and S. aureus isolated from food.

2. Materials and Methods

This experimental study was performed on two pathogenic and spoilage bacteria in meat and vegetable foods along with two standard strains of the same bacteria. In order to use the collected isolates, they were stored in a tryptic soy broth containing 15% of cultured glycerol at -70° C. The standard strains of P. aeruginosa (ATCC 27853) and S. aureus (ATCC 25923) used in this study were purchased from Zistroyesh Company in a freeze-dried form. In order to prepare the bacterial suspension for daily tests, McFarland suspension (1-1.5×108 mL) was prepared. To ensure the correct turbidity of the McFarland suspension, its absorption was measured by a spectrophotometer at a wavelength range of 620 nm [17].
First, 100 g of zeolite with 70 g of zinc acetate was poured into 500-m beaker and, then, 400 mL of deionized water was added to them. This beaker was then placed on a magnetic steering device. After 30 minutes, the beaker content was filtered using Whatman cellulose filter paper (Grade 40) and a white filter band (S&S 589/2: 12-25 μm, Germany) and washed by 500 mL of deionized water. Both filtered contents were then transferred to a glass plate and dried for 24 hours at room temperature. On the second day, the plate was incubated for 80 hours at 80° C and then was placed in a 120° C oven for 2 hours. The 400° C furnace was then used for calcinations of the obtained material for 2 hours [18]. Finally, to measure the amount of ZnO, the XRF analyzer (PW 2404, Philips Co., Holland) was used available at the laboratory of Tarbiat Modares University.
The lowest concentration of nanoparticle suspension that did not show turbidity in the tube was determined as the Minimum Inhibitory Concentration (MIC) of nanoparticle growth. In tubes with no growth, Minimum Bactericidal Concentrations (MBC) was determined by performing a re-culture on Müller-Hinton agar medium. These assessments were repeated three times [19]. All mediums used in this study were prepared from Merck Company in Germany. The antibacterial effect of ZnO nanoparticles on standard strains of P. aeruginosa and S. aureus isolated from food was evaluated by macrodilution method which includes the determination of MBC and MIC values and disk diffusion.

3. Results

Using the XRF analyzer, different percentages of elements in non-nano ZnO suspension and ZnO nanoparticle suspension were determined. The amount of ZnO non-nano ZnO suspension was obtained 8.358 and the amount of ZnO nanoparticle suspension was 25.149. The MIC of ZnO nanoparticle growth was reported 4 mg/mL for the standard strain and isolate of P. aeruginosa and 2 mg/mL for the standard strain and isolate of S. aureus. The MBC of ZnO nanoparticle suspension for the standard strain and P. aeruginosa was obtained 16 and 8 mg/mL, respectively, and for the standard strain and isolate of S. aureus was 8 mg/mL

4. Conclusion

The results of this study showed that ZnO nanoparticle suspension had better antimicrobial effects on all bacteria compared to zeolite (non-nano ZnO) . During this study, antibacterial activity increased with the increase in the concentration of the nanoparticle solution. Reddy et al. examined the antimicrobial effects of ZnO nanoparticles on S. aureus and Escherichia coli and found that gram-positive S. aureus was more sensitive to ZnO nanoparticles than gram-negative Escherichia coli, which is consistent with the results of our study [24]. Ramani et al. synthesized ZnO nanoparticles with different structures and examined its antibacterial properties on 4 strains of gram-positive bacteria and 4 strains of gram-negative bacteria, and observed that spherical ZnO nanoparticles had better antibacterial properties [24]. Seil et al. synthesized a composite of polyvinyl chloride and ZnO nanoparticles and studied its antibacterial effect on S. aureus and showed that ZnO improves the antibacterial properties of the study composite [25]. According to the results of the present study, it can be found that S. aureus was more sensitive to ZnO nanoparticles than P. aeruginosa. 
It can be concluded that ZnO nanoparticles can be used in food packaging and storage as deterrents to pathogenic bacteria and food spoilage, leading to reduced consumption of raw materials and less waste in the packaging industry. The isolates of bacteria were more sensitive than the standard strains. Standard strains were clinical samples and become resistant over time due to the use of antibiotics.

Ethical Considerations

Compliance with ethical guidelines

This study obtained its ethical approval from the Ethics Committee of Tehran University of Medical Sciences (Code: IR.TUMS.VCR.REC.1397.484).

Funding

This study was extracted from the master thesis of first author, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences (Code: 39078).

Authors' contributions

Data collection, experiments, editing & review: Shima Naddafi, Zahra Dargahi; Data analysis and interpretation: Mohammad Mehdi Soltan Dallal, Alireza Partoazar; Initial draft preparation: Shima Naddafi. 

Conflicts of interest

The authors declare no conflicts of interest.
 

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Type of Study: Research | Subject: Microbiology

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