Study the effect of antibiotics and propolis on pathogenicity of the methicillin resistant staphylococcus aureus (MRSA)‎

Other Title(s)

دراسة تأثير المضادات الحيوية و العكبر على إمراضية بكتريا المكورات العنقودية الذهبية المقاومة للمثسلين

Dissertant

al-Shaykh, Ilaf Basim Nuri

Thesis advisor

Yusuf, Hana Salim

Comitee Members

Alash, Samir Abd al-Amir Abd Ali
Abbud, Rana Sadi
Isa, Rajwa H.

University

University of Baghdad

Faculty

College of Science

Department

Department of Biology

University Country

Iraq

Degree

Master

Degree Date

2013

English Abstract

This study included collection of 100 swab specimens from patients in AL-Kindy Teaching Hospital and teaching laboratories of Medical City Hospitals in Baghdad during the period from August to December 2012 , these swab specimens differed in their sources which included 19 nasal swab, 16 wound swab, 27 burn swab, 7 pus swab, 15 sputum swab, 10 corneal swab and 6 urine swab .

Only 38 (38%) isolates were identified as Staphylococcus aureus, 38 isolates of them (82.6%) were coagulase-positive (COPS), while 8 isolates(17.3%) were coagulase negative (CONS), from total 46 isolates of Staphylococci.

The distribution of Methicillin resistance among Staphylococcus spp isolates was investigated by disc diffusion method.

In this study, 21 isolates (55.26%) out of 38 (38%) isolates were identified as Staphylococcus showed to be resistant to the Methicillin while 17 isolates (44.73%) were sensitive.

The highest rate of Methicillin resistance Staphylococci were obtained from wound and pus swabs.

The results obtained from antimicrobial sensitivity test showed that MRSA isolates were resistant to many other antimicrobial agents in addition to the Methicillin (Multi-drug Resistant).

Antimicrobial activities of different antimicrobial agents include (crude Ethanolic Extract Propolis (EEP)), Lysostaphin, Ciprofloxacin and Vancomycin) were tested singly and in combination against selected isolates (MRSA S3) by using agar disc diffusion assay.

Results revealed that S.

aureus (MRSA- S3) was more sensitive to 2 μg/ml concentration of EEP than other antimicrobial agents used in this study.

However, antibacterial activity of the other three reagents were significantly less than propolis, where MRSA isolate showed resistance for their low concentrations.

In comparing the pharmacodynamics of lysostaphin, vancomycin and Ciprofloxacilin against selected isolate (MRSA -S3) there was no significant differences with (P>0.05) between those drugs at the diameter of inhibitions zone which were (11, 11and 10mm) respectively, the effected concentration of them were (5.625μg/ml,16μg/ml and 3%).

The results showed a synergistic effect of this combination on the selected isolate (MRSA- S3), as well as, same results obtained when combined propolis with ciprofloxacin.

The bacterial growth was inhibited with elevation of inhibition zone to (11, 13mm) respectively, while selected isolate (MRSA-S3) showed ahigh resistance against the combination of vancomycin with (propolis and lysostaphin ) ,so they effects was antagonistic to S.

aureus.

The ability to produce slime layer by MRSA isolates was also investigated and the results showed that all isolates of MRSA have produced a slime layer when tested by tube method, but the amounts of adherent materials were differ among the isolates.

However, the results by Congo red agar method showed that 57% of MRSA isolates produced strong slime layer and 43% of MRSA revealed negative result.

Similarly the ability of MRSA to produce biofilm by tissue culture plate (TCP) was investigated and the results indicated that MRSA isolates showed strong ability to form abiofilm , and the OD value of biofilm formation ranged between (0.262 - 0.311nm).

moreover the OD value of biofilm formation significantly increased after addition of 1% glucose to the media.

The Antimicrobial activity of propolis and lysostaphin on the biofilm formed by MRSA isolates were investigated and the results showed that propolis at MIC (2μg/ml) were significantly inhibited the biofilm formation by MRSA S3 isolates when used alone, the optical density significantly (P<0.05) decreased to (0.027nm) in comparison with control group (0.3875nm).

Statistical analysis showed a slightly effect of lysostaphin under therapeutic concentration (5.625μg/ml ) on biofilm formation ability of (MRSA- S3) optical density was reduced to (0.312 nm) in compared to control group (0.389nm).

On the other hand, the results showed that the synergistic combination of propolis with (lysostaphin, ciprofloxacin) at therapeutic concentration were inhibited the biofilm formation by selected isolates.

Histo-pathological effect of different concentrations of propolis, lysostaphin and ciprofloxacin on the keratitis treatment induced in the rabbit eyes was studied.

Organs of rabbits (eyes) showed different pathological changes in a corneas treated with 2μg/ml propolis alone were significantly less inflamed than untreated MRSA S3–infected corneas.

The corneal epithelium and endothelium looked like an uninfected cornea, clinical signs of keratitis in the experimental rabbit were completely disappeared with score 0: they had normal and healthy appearance compared with negative control group (uninfected eyes).

Corneas treated with lysostaphin were highly edematous in comparison to uninfected eyes.

Histopathological changes showed highly neutrophils infiltrations with highly damaged endothelial layer, corneas treated with lysostaphin had similar appearance to untreated corneas.

On the other hand, when combined propolis with lysostaphin and propolis with ciprofloxacin results induced amoderate recovering in the infected eyes, the pathological changes included moderate corneal ulceration, conjunctively edema, and minor accumulation of fibrin in the anterior chamber were the primary changes noted in those treated groups, than untreated corneas.

While when compared the current results with a healthy cornea, it was found that the infected group still inflamed after treated with reagents combinations, also there’s significant (P<0.05) difference in comparable to un treated eyes.

Main Subjects

Biology

No. of Pages

181

Table of Contents

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Language

English

Data Type

Arab Theses

Record ID

BIM-597854