Bioactivity against Staphylococcus aureus of New Nanoparticles Derived from Iron Oxide and Beta vulgaris Extracts and Synthesis Characterization

 

Manal Khalid Mohammad, Sundus Hameed Ahmed, Rana Fadhil Abbas

Biology Department, College of Science, Mustansiriyah University.

*Corresponding Author E-mail: manalalkarhi@gmail.com

 

ABSTRACT:

In this study an opportunity was obtained that combines the VITEK system and the detection system of clinical microbiology laboratories to reach the rapid microbial identification and sensitivity test for Staph aureus bacteria, where the results of the susceptibility test were compared to 3925 combinations of antibiotics. The total rate was for the normal antimicrobials as well as the category approval for direct sensitivity test. 92% with less than 3% significant errors and the average time required to identify the identity with a sensitivity test is 7.5 hours (3.0 SD hours), compared to an average of 32.3 hours 9-14 hours for the normal method. The study focuses on the effectiveness of anti-bacterial iron nanoparticles from Beta vulgaris extract in various concentrations (1 mg/ml. 2 mg/ml. 3 mg/ml.4 mg/ml. against Stphylococcus aureus. Where in an inhibition zone was obtained for the highest concentration of iron nanoparticles for Beta vulgaris when compared to other samples.

 

KEYWORDS: Iron Nanoparticles, Beta vulgaris, Stphylococcus aureus, VITEK system.

 

 


INTRODUCTION:

Staphylococcus aureus is a Gram-positive, round in shaped, and considered as microbiota of the body, that found on the skin the upper respiratory tract. It is facultative anaerobe that grows without oxygen also positive for catalase and nitrate reduction 1. Although S. aureus due to as a commensals of the human microbiota and also become an opportunistic pathogen, can cause of infections to skin including respiratory infections such as sinusitis, abscesses, and food poisoning. virulence factors producing from pathogenic strains like protein toxins, and inactivates antibodies. by cell-surface protein methicillin-resistant S. aureus (MRSA) is the emergence of antibiotic-resistant strains of S. aureus that cause large problem in clinical medicine. It does not vaccine for S. aureus has been done. It considered as normal flora in skin in nostrils2 and normal of the lower reproductive tract for women and can cause skin infection like impetigo, folliculitis, boils, cellulitis, scalded skin syndrome, carbuncles It is stay one from five most common infect of hospital-acquired infections and cause of wound infections following surgery2,3.

 

Recently, iron oxide (FeNPs) considered as a nanoparticles it becomes the attention and the consideration of authors, due to its exceptional properties, such as surface-to-volunie, super paramagnetism, easy separation and its greater surface area in addition to its bioactivity against microbes. The synthesis of iron oxide nanoparticles from plant materials offers several benefits like eco-friendliness and a wide and various applications because the synthesis protocol do not involve toxic chemicals4 system is found in three types (VITEK 2 XL, VITEK 2 compact and VITEK). The an automatic microbiology system is VITEK 2 for technology of utilizing growth-based. All the types of system have the same colorimetric reagent cards its mean similar in a commodation. That automatically interpreted and incubated. The VITEK 2 was used in   this study by researcher5,6.

 

The study aimed to synthesize iron nanoparticles from iron oxide and Beta vulgaris extract, characterized them and test its bioactivity tracts and test its antimicrobial activity against Staphylococcus aureus.

 

METHODOLOGY:

Biosynthesis of Iron nanoparticles:

Beta vulgaris is the fresh fruits of crop were locally collected. Then fruits were washed with tap water for several times to remove the contaminant, after that, 10 g of the fruit was taken, cut into pieces as small, immersed in 100 rnL of distilled water and heated up for 30 min to 80 °C Then was filtered the hot solution to get the Beta vulgaris extract. The extract was stored in a refrigerator at 4 °C for 24 hrs. After that. 10 ml. of fruit extract was added dropwise to 0.01 M aqueous solution of Iron oxide. The mixture was kept with continuous mixing for 10 min to get collides and a dark black colour visually observed which medicates the formation of iron nanoparticles (FeNPs) P 7.

 

Diagnosis of Staphylococcus aureus by using of Vitek®2 compact:

Staphylococcus aureus cultured on media called nutrient agar for isolates and to obtain colonies in a single were incubated for 24 hrs at 37 °C, then identified the isolates after growth of colonies by using Vitek'®2 compact. The system manufactured by (BioMeneux, USA), the analysis was earned out m Public health lab in Najaf as suggested by Darbandi in 2010 and confirmed by Yahya and Ghosoon in 2016 8,9.

 

Antibiotic susceptibility test:

Kirby-Bauer disc diffusion method was used for Antimicrobial susceptibility’ testing on Mueller- Hinton agar that recommended through the Clinical Laboratory Standard Institute (10). The antimicrobial sensitivity results and their corresponding concentrations are shown in table 1.    

 

Table 1: Antibiotic sensitivity and correspond concentrations to    Staphylococcus aureus

No.

Compound

Concentration

1.

Amikacin

10 mg

2.

Ciprofloxacin

10 mg

3.

Gentamicin

10 mg

4.

EDTA

980 mg

5.

Ampicillin

25 mg

6.

Cloxacillin

5 mg

7.

Trimethoprim

1.25 mg

8.

Suphamethoxazole

23 75 mg

9.

Vancomycin

30 mg

10

Imipenem

10 mg

11.

Nitrofurantoin

100 mg

12

Rifampicm

5 mg

 

Anti Bacterial:

The bacterial suspension with moderate turbidity was inoculated on Mueller-Hinton plates and was incubated for 18-24 hrs at 37 °C, the resistance and the susceptibility of the Staphylococcus aureus. was measured for all antimicrobial agent with four different FeNPs concentrations (1, 2, 3, and 4 mg/I) as well as the fruit extract, and Iron oxide, the resides were interpreted according to MacFaddin 2000 criteria 11.

 

RESULTS AND DISCUSSION:

Table 2 shows Vitek 2 results for Staphylococcus aureus the compact system that was used had several advantages, it become identified with in 6 hrs comparing with other methods considered traditional that need 2-3 days, besides that it was automation in a high level of taxonomically and simple methodology. The test was carried out as explained in 2017 by Bhalerao and          Borkar 12.


Table 2: Vitek 2 results lav Staphylococcus aureus

Biochemical Details

2

AMY

-

28

AlaA

-

57

Draf

-

4

PIPLC

-

29

TyrA

-

58

O129R

+

13

APPA

-

38

dRIB

-

12

AGLTp

-

14

CDEX

-

39

ILATK

+

5

AspA

-

20

LeuA

--

47

NOVO

-

64

OPTO

+

23

ProA

-

50

NC6.5

+

24

BGURr

-

30

dSOR

-

16

BGAR

-

60

SAC

+

45

dMAL

+

27

BGUR

-

 

 

-

42

LAC

-

25

AGAL

-

9

BGAL

+

54

MBdG

+

37

dGAL

-

 

 

-

52

dMAN

+

31

URE

+

17

AMAN

-

62

dTRE

+

46

BACI

+

 

 

 

-

59

SAL

-

44

NAG

+

26

PyrA

+

11

AGLU

-

56

PUL

-

 

 

 

8

ADH1

+

53

dMNE

+

32

POLYB

+

19

PHOS

+

63

ADH2s

-

 

 

 

 


The antibiotic sensitivity test for Staphylococcus aureus shows high sensitivity against Gentamicin, Ciprofloxacin, Amikacin, Nitrofurantoin. EDTA. and Imipenem, while it exhibit resistant activity against Vancomycin, Rifampicin. Ampicillin and Cloxacillin as shown in Table 3

 

Table 3: Antibiotic Sensitivity

No.

C oinpound

Concentration

Sensitivity

1.

Amikacin

10 meg

S

2.

Inupenem

10 meg

S

3.

Gentamicin

10 meg

S

4.

Vancomycin

30 meg

R

5.

Ampicillin

25 meg

S

6.

Rifampicin

5 meg

R

7.

Trimethoprim

1.25 meg

I

8.

Suphamethoxazole

23.75 meg

I

9.

EDTA

980 meg

S

10.

Ciprofloxacin

10 meg

S

11.

Nitrofurantoin

100 meg

R

12.

Cloxacillin

5 meg

R

I is Insurance, R is Resistance, and S is Sensitivity

 

The color of non nanoparticles (FeNPs) solution was black, w’hile the non oxide solution was yellow, and the Beta vulgaris extract most like brownish read, the difference m colors is shown in figure 1.

 

Figure 1: Color changes of A)- FeNPs; B)- Iron Oxide; C)- Beta vulgaris extract

 

UV-Vis Spectra was carried to FeNPs solution: it shows a peak at 380-420 which confirm the coordmation between the orgasmic extract with iron ion and the formation of FeNPs. Also, color changing can be considered as a confirmation of the formation of iron nanoparticles. Methods which utilize biological materials such as plants prove an effective way for the nanoparticles synthesis, when use the plants possess many benefit like, easy scale-up for large-scale synthesis and cost-effectiveness. Besides in nanoparticles synthesis, plant extracts make both as reducing agents and stabilizing agents.

 

Figure 2 shows the inhibition zone of Staphylococcus aureus. by nanoparticle. Extract, oxide and D.W. while figure 3 shows the antibacterial activity against Staphylococcus aureus. for FeNPs. non oxide and Beta vulgaris extract. The test was carried out using four concentrations (1 mg/ml. 2 mg/ml. 3 mg/ml and 4 mg/ml) to each one. It was found that the inhibition zone increased with the increases of concentrations. The inhibition zone for 1 mg ml was 0.8 cm. for 2 mg/ml was 1.2 cm. for 3 mg/ml was 1.9 cm. and for 4 mg/I were 2 cm.

 

Figure 2: The inhibition zone of solutions

 

Fig 3: Antibacterial activity of: 1. Iron oxide nanoparticles 2. Beta vulgaris extract; 3. Iron Oxide

 

The results in this research medicate the high bioactivity of FeNPs against Staphylococcus aureus comparing with iron oxide and Beta vulgaris extract, the researcher recommends testing the bioactivity of iron nanoparticles against different types of microbes and even cancer cell lines that affect on humans and animals. Also, the researchers recommend using Beta vulgaris extract for the synthesis of different metal nanoparticles like copper, titanium, cobalt, zinc, cadmium and test their nanoparticles bioactivities.

 

ACKNOWLEDGMENT:

The authors would like to thank Mustansiriyah University (www. uomustansiriyah.edu.iq) Baghdad-Iraq for its support in the present work.

 

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Received on 15.06.2021           Modified on 12.02.2022

Accepted on 06.06.2022         © RJPT All right reserved

Research J. Pharm. and Tech. 2022; 15(7):3169-3172.

DOI: 10.52711/0974-360X.2022.00530