Antibacterial action of Silver Nanoparticles against Staphylococcus aureus Isolated from wound infection
Haider Qassim Raheem1, Ehasn F. Hussein2, Ahmed Hameed Rasheed3, Najwan K. Imran4
1,3,4DNA Research Center, University of Babylon, Hilla, Babylon, Iraq.
2College of Medicine of Hamorabi University of Babylon, Hilla, Babylon, Iraq.
*Corresponding Author E-mail: haiderbio412@gmail.com, ehsan.f.hussein@gmail.com, alhly612@gmail.com, najwankadhim.imra@uobabylon.edu.iq
ABSTRACT:
This study aimed to assess antibiotics resistance and antibacterial action of silver nanoparticles against Staphylococcus aureus isolated from wound infection. A total of 100 samples of wound swab existed calm since wound patients who stayed the Al-Hillah, Teaching Hospital (wound unit) in, Babylon province, Iraq. S aureus was recognized biochemically and morphologically. A whole of 30(30%) of the whole specimens tested confident for S.aureus. Available of 30 S.aureus isolates, 8(26.6 percent) were MRSA. Antibiotic susceptibility for 8 antibiotics for MRSA that appeared to Penicillin G and Cefoxitin was tested, and all isolates were resistant (100percent), Were susceptible to Rifampin, Tetracycline, and Ciprofloxacin (62.5percent) Clindamycin sensitivity remained experimental in 75% isolates. Resistance to Erythromycin remained establish in approximately 62.5 percent of the population. Gentamycin resistance was found in 50% of the isolates. The antibacterial activity of silver nanoparticles (AgNPs) alongside S.aureus demonstrates extreme broad-range antibacterial act in contradiction of recognized bacteria, with an rise inhibition zone diameter related to nanoparticle concentration The MIC of Ag NPs ranged from 50 to 100g/ml, while the MBC ranged from 100 to 200g/ml. Ag NPs is suggested as an effective anti-MRSA alternative. This experiment discovered that Ag NPs is highly recommended as an alternative anti-MRSA agent with significant inhibitory and antibacterial effect due to the methicillin resistant strains' high rate of resistance to Penicillin G and Cefoxitin (100%), Erythromycin (62.5%), and Gentamycin resistance (50%).
KEYWORDS: Antibacterial, Ag, Nanoparticles, S. aureus, MIC, MBC.
INTRODUCTION:
Here numerous organic in addition chemical composites thru anti-bacterial action, for example penicillins besides normal produces, which eradicate bacteria or dawdling their growing. Nanoparticles, for example, require newly received increased consideration.1 Reactive oxygen species (ROS) for example, organic hydroperoxides, hydrogen peroxide hydroxyl radicals, also superoxide NPs statement scheduled the external of bacteria, then NPs gathering in the cytoplasm (periplasmic)area of bacteria canister stay a clue toward bacteria kill.3
Trendy the example of bacteria, (ROS)can cause the destruction of components of cell such as peptidoglycan, lipids, DNA also proteins by producing ROS via NPs4. Silver nanoparticles (NPs) are important components for research in the field of nanomedicine. This focus is based on physical-chemical factors such as shape and size. The external area to size percentage of NPs is critical intended for things.5,6 In this instance, one of the most significant pragmatic ingredients in manufacturing is silver (Ag), then its compound in nano-meters is the rule8,9. These silver nanoparticles canister be used as a good substitute for gold and copper nanoparticles10,11 Among several metallic oxide NPs, Ag has received the most attention because it is the greenest member of the intimate of silver composites and exhibits a variety of beneficial physical properties such as exceptional heat conductivity, electron link special effects, and gyration diminuendos.12,13,14
S.aureus is a Gram-positive pathogen that causes severe infections and diseases that result in significant illness and death. The development of novel (drug resistant) straining of this bacteria for example a outcome of uncontrolled use of antibiotic is currently a major source of concern.15 S.aureus is individual of the furthermost dangerous Staphylococci spp faced. Myocarditis, osteomyelitis, bacteremia, pneumonia, pericarditis, meningitis, encephalitis, mastitis, and scalded skin syndrome are all caused by it.16 Methicillin-resistant) S.aureus MRSA strains, also known as multidrug-resistant S.aureus, emerged in the latter period as a source of hospital infections linked to rapidly advanced latent fatal diseases for example life-threatening fasciitis, pneumonia, endocarditis, necrotizing fasciitis, osteomyelitis, sepsis and (toxic shock syndrome), For the ever-increasing number of life-threatening resistant strains, a safe and efficient alternative is desperately needed.17
MATERIALS AND METHODS:
Sources of materials:
Culture media and Antibiotics discs used in this study are listed.in table (1,2):
Table 1: Culture media used in this study.
|
Media |
Origin |
|
Nutrient agar, Nutrient broth, Mannitol salt agar, Ames transport, Blood agar base, Muller-Hinton agar and Brain heart agar |
Himedia -India |
Antibiotics discs were purchased from (Bioanalyse /Turkey) and are listed below.
|
Antibiotic Discs potency (µg) |
Symbol |
|
Tetracycline 30 |
TE |
|
Erythromycin15 |
E |
|
Gentamycin 10 |
CN |
|
PenicillinG 10 |
PCN |
|
Cefoxitin 30 |
CF |
|
Rifampin 5 |
RA |
|
Clindamycin 30 |
DA |
|
Ciprofloxacin 15 |
CIP |
Specimen Collections and Isolation:
Bacterial isolates were obtained as of patients by the Al-Hillah, Teaching Hospital (wound unit) in, Babylon province, Iraq. A total of 100 specimens were collected using Ames transport media, dwindling hooked on brain heart infusion broth (BHI), then incubated at (37°C for 24hr). Mannitol, Salt, Agar plates were inoculated then incubated at (37°C for 24hr). to isolate S.aureus .On nutrient agar, isolated colonies were purified (NA). According to Bergey's manual, S.aureus was recognized based on morphological features and biochemical tests on culture media.18
Antimicrobial Susceptibility Test by Disk diffusion Method:
The antibiotic names and standard inhibition diameters were used in this test, as recommended by19. The Inhibition zone dimeter was measured with a caliper and compared to those determined through the (Clinical Laboratory Standards Institute, CLSI 2021) to determine antibiotic resistance (R) or sensitivity (S).
Identification of Methicillin Resistant S. aureus:
Cefoxitin (30g) disc was used as a substitute for Methicillin, inhibition zone diameter of (19mm) was considered resistant to Methicillin, and inhibition zone diameter of (20mm) stayed reflected sensitive.20
Antibacterial activity of Ag NPs:
Ag NPs (40nm) were obtained from (US research nanomateril as Inc USA). The CLSI-specified antibacterial action was administered. As previously stated, the disk diffusion method is castoff to measure bacteria sensitivity to Ag NPs15. Triplicates of Ag NPs in deionized water were prepared (400, 200, 100, 50, 25, and 1.25g/ml), and ready discs were emerged in each dilution before usage. The MRSA isolates stayed incubated at 4°C for 15 minutes beforehand to 37°C for 24hours. After detecting a zone of inhibition, the positive results were documented.21
Minimum bactericidal concentration (MBC): determination) and Minimum inhibitory concentration (MIC):
Bacterial isolates were incubated overnight at 37°C and used to determine 0.5 McFarland. In triplicate, 10ml tubes of nutrient broth medium were prepared with five dilutions of Ag NPs (400, 200, 100, 50, 25, and 1.25g/ml) and a negative control (without Ag NPs). Each tube was (inoculated)with 1ml of the suspension of bacteria. The inoculated circles remained incubated overnight at 37°C. The MIC is defined such as the lowermost concentration through nope turbidity. Tubes with nope turbidity stayed cultured overnight proceeding nutrient agar at 37°C. Bacterial colony growth was observed, then the concentration that shows no growing is denoted as the MBC.22,23
RESULTS:
Thirty (30%) of the 100 wound swab samples used in the study tested positive for S. aureus in culture. Only 8 (26.6 percent) of the isolates were found to be MRSA.MRSA had the highest level of susceptibility (62.5percent) to Rifampin, Ciprofloxacin and Tetracycline, while 37percent were resistant. Completely isolates remained completely resistant to Penicillin (G), Cefoxitin. (25%) were Clindamycin resistant.
Table 3: Antibiotic Susceptibility Patterns and Antibacterial activity of AgNPs on S.aureus
|
Antimicrobial agent |
Inhibition zone diameter(mm) |
||||||||
|
No(1) |
No(2) |
No(3) |
No(4) |
No(5) |
No (6) |
No(8) |
No(8) |
|
|
|
Tetracycline |
12(R) |
13 (R) |
20 (S) |
21(S) |
21(S) |
20(S) |
13 (R) |
21(S) |
|
|
Gentamycin |
18(S) |
10(R) |
13(S) |
11(R) |
17(S) |
11(R) |
11(R) |
15(S) |
|
|
Erythromycin |
12(R) |
23(S) |
13(R) |
11(R) |
13 (R) |
23(S) |
12(R) |
23(S) |
|
|
Rifampin |
20 (S) |
15(R) |
21(S) |
14(R) |
21(S) |
21(S) |
15(R) |
21(S) |
|
|
PenicillinG |
19 (R) |
21(R) |
20(R) |
19(R) |
18(R) |
21(R) |
22(R) |
21(R) |
|
|
Cefoxitin |
20(R) |
18(R) |
18(R) |
19(R) |
19(R) |
17(R) |
18(R) |
19(R) |
|
|
Clindamycin |
20(S) |
19(S) |
20(S) |
22(S) |
14(R) |
13(R) |
18(S) |
20(S) |
|
|
Ciprofloxacin |
23 (S) |
13(R) |
23(S) |
22(S) |
13(R) |
22(S) |
14(R) |
23(S) |
|
|
Ag 400μg/ml |
22 |
23 |
20 |
24 |
22 |
23 |
22 |
24 |
|
|
Ag 200μg/ml |
20 |
18 |
18 |
20 |
21 |
20 |
19 |
21 |
|
|
Ag 100μg/ml |
18 |
17 |
16 |
18 |
16 |
18 |
19 |
17 |
|
|
Ag 50μg/ml |
14 |
15 |
15 |
16 |
14 |
16 |
!6 |
13 |
|
|
Ag 25μg/ml |
12 |
11 |
10 |
9 |
11 |
10 |
12 |
11 |
|
|
Ag 12,5μg/ml |
8 |
6 |
7 |
5 |
6 |
8 |
6 |
7 |
|
-R: Resistant. –S: Sensitive
Figuer 2: Antibacterial activity of Ag NPs on S. aureus (MRSA).
Erythromycin resistance was found in approximately 62.5 percent of the population, while 37.5 percent were sensitive. Gentamycin resistance was found in approximately half of the isolates (table 3). (figure 1). With MRSA, AgNPs exhibits a wide range of antibacterial activity The results in table (3) demonstrated the action of Ag NPs, which showed an increase of inhibition zone diameter relative to the rise in concentration of nanoparticle. Figure (2) shows that the MIC of AgNps ranged from 50g/ml to 100g/ml and the MBC ranged from 100g/ml to 200g. Figure (3) shows that the MIC of AgNps ranged from 50g/ml to 100g/ml and the MBC ranged from 100g/ml to 200g.
Figure 3: MIC and MBC of Ag NPs on S. aureus (MRSA).
DISCUSSION:
This finding contradicts the findings of. 24,25, who discovered that Erythromycin, Tetracycline, Gentamycin, Ciprofloxacin, and Rifampin have resistance percentages of 37.7 percent,34.9 percent, 28.5 percent, 29.2 percent, and 3.8 percent, respectively, but agree with the rates of resistance to Penicillin G and Cefoxitin (100%), as well as Clindamycin (32.8%). In addition26discovered that 60 percent of the isolates were Clindamycin resistant. These variations in results could be attributed to differences in topographical zones, isolate quantities, and gathering locations.
AgNPs exhibits a wide range of antibacterial activity this finding is consistent with the findings of20, who discovered that silver nanoparticles are antibacterial against S.aureus. In addition to superoxide species existence made and causal to the breakdown of biomolecules, AgNPs caused the discharge of reactive oxygen species (ROS), a fast drop in bacterial cell membrane integrity.27
CONCLUSION
This experiment discovered that Ag NPs is highly recommended as an alternative anti-MRSA agent with significant inhibitory and antibacterial effect due to the methicillin resistant strains' high rate of resistance to Penicillin G and Cefoxitin (100%), Erythromycin (62.5%), and Gentamycin resistance (50%).
CONFLICT OF INTEREST:
The authors declare no conflict of interest.
ACKNOWLEDGEMENTS:
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Received on 29.09.2021 Modified on 03.11.2021
Accepted on 06.12.2021 © RJPT All right reserved
Research J. Pharm. and Tech. 2022; 15(6):2413-2416.
DOI: 10.52711/0974-360X.2022.00401