Antibacterial activity of garlic against clinical isolates of staphylococcus aureus

 

Antibacterial activity of garlic against clinical isolates of staphylococcus aureus

Introduction

Garlic  (Allium sativum L.) is under family Liliaceous. It is an erect annual herb with superficial adventitious roots, bulbs composed of a disk like stem (Abebe D, 2003). It has long tradition as medicinal plant, started with a direction of preparing a medicinal remedy written in a cuneiform character in about 3000 BC. Scientific investigations of various garlic preparations began in 1939 (Weiss RF, 1988)

The use of higher plants and preparations from them to treat infections is an age-old practice. Interests in plants with antimicrobial properties have come to use again because of emergence of resistance strains against antimicrobials such as penicillin (Dikasso D, 1999). There are a number of studies carried out to assess the value of herbal remedies including garlic preparations for treat of illness.

This study will confirm the aqueous extract of Garlic had antibacterial effect against clinical isolates of S. aureus.

(Ruqiya Sania)

Statement of problem

The hospital infection (Staphylococcus aureus) is at the top of major diseases,the organism we will use for our study is Staph. aureus which is resistant towards MARSA, we will remove it by using Garlic extract because Garlic has antibacterial activity.


Hypothesis

Garlic  extract has an antibacterial  properties against Staphylococcus aureus , our prediction is to show 80-90 % results against staph. infection.


Key words

Lab: Laboratory; MRSA: Methicillin Resistant Staphylococcus aureus; MSA: Mannitol Salt Agar; Staph: Staphylococcus. GPGCM: Govt. postgraduate college Mandian


 

Objectives

 

1.      To obtain the activity of garlic against nosocomial infection

2.      To evaluate the  anti-bacterial effect of garlic against clinical and standard isolates of S. aureus


Literature Review

Garlic as an antibiotic

Garlic is an antibacterial agent that naturally inhibits growth of infectious agents and at same time protects the body from the pathogen .It is known that the most sensitive bacterium to garlic is the deadly bacillus anthraces which cause the diseases anthrax.

 Allicin

Allicin is antibacterial components found in garlic . A molecular may be the basis for some of garlic , therapeutic effect .The researchers were able to study how  garlic works at molecular level using allicin , garlic's main biologically active components. Allicin created when garlic cloves are crushed , protects the plants from soil parasites and fungi and is also responsible for garlic's pungent smell.

It is natural weapon against infection that disables dysentery causing anabasis by blocking two groups off enzymes, cysteine proteinases and alcohol dehydrogenases. Cysteine proteinases enzymes are main culprits in infection, providing infectious organs with means to damage and invade tissue . Alcohol dehydrogenase enzymes play major role in these harmful organism survival and metabolism. Because of these groups of enzymes found in wide variety of infectious organism such as bacteria, fungi , and viruses . This research provide scientific bases motion that allicin is broad spectrum antimicrobial, capable of warding off different type of infectious .

It is likely that bacteria would develop resistance to allicin because this would require modifying very enzymes that make their activity possible. Scientist found that allicin blocks enzymes by reacting with one of their important component known as self hydl(Sh) group , or throts this finding has important implication because of sulfhydryl groups O are also crucial component of some enzymes that participate in synthesis of cholesterol . “GARLIC LOWERS THE LEVEL OF HARMFUL CHOLESTEROL”.

 Review on test organism

Staphylococcus aureus is very important pathogen that cause variety of disease including skin infections. Gastrointestinal diseases, food poisoning, toxic shock syndrome and nosocomial infection acquired during hospitalization.

Staphylococcus aureus is isolated on Mannitol salt agar which is medium that is high in salt (75%) and contain Mannitol as source of carbon as energy. S.aureus ferments the mannitol and cause medium turn yellow.


 Disclosure of Methicillin Resistant Strains of S. aureus (MRSA)

 

There is a growing medical problem due to increasing frequency of infection caused by penicillinresistant Staphylococci. B-lactamase producing strains of S. aureus that are resistant to penicillin first appeared in clinical specimens in early 1950s. Soon thereafter, multiple antibiotic resistances were detected in chemical isolates of S. aureus; these strains were resistant to macrolide antibiotic, amino glycoside, and tetracycline.


Materials and Methods

Plant materials

 

We will obtain the Garlic solution not from the whole part of the plant rather on the bulbs. The bulb of Garlic will peel off, weighted and then ligated using pestle while adding small amount of H2O. The extracts will then allow freezing at -18°C (deep freezing) so as to concentrate the chemical (allicin). Then after freezing, we will put filtrate into lyophilizer till an amorphous powder weighted and then diluted with distilled water and used for the experiment

Test organism

Test organism (S. aureus ) will isolate clinically from patients which will collected from Awasssa Referral Hospital.

 standard pure culture will  collected from EHNRI (Ethiopian Health and Nutrition Research Institute). Clinical and standard will sub cultured into subsequent Nutrient broth and definite Medias which is favour i.e., Mannitol salt agar (MSA) for S. aureus will prepare on a slant and on petri dishes.

Inoculum

Cultured of the test organisms will maintained on nutrient. Briefly, we will pic four to six colonies were picked with an inoculating loop and suspended in 5 ml of broth and incubate at 37°C for 24 hours. The turbidity of the broth culture was then equilibrated to match that of 0.5 McFarland standards. This provides organisms in the range of 1 × 106 to 5 × 108 cfu/mol which is pathogenic that used for the test

Antibacterial activity test

The antibacterial activity test of the crude extract of Garlic against both standard and clinical isolates will be carried out by the Agar diffusion method.

Agar diffusion method

The molten agar will mixed with a different concentrations of the test samples at molten state 45-50°C and mixed aseptically with different amounts of garlic extracts to a concentration of 0.25 ml, 0.5 ml, 0.75 ml and 1.5 ml which is equivalent to 5 mg/ml, 10 mg/ml, 15 mg/ml and 25 mg/ml of media. Then, the prepared media will let to solidify. A separate agar plate without sample or drugs will also be prepared in order to provide an appropriate growth of organisms. (As the same time as control) Two standard drugs as a positive control will also test against these microorganisms. These will chloramphenicol 0.30 mg and penicillin 0.30 mg. The negative control used in the cork borer as well as the solvent i.e., distilled water.

Antibacterial effect will be determined by direct visual comparison of the growth of the test cultures. All the tests will carried out in triplicate and the results will be reported as the averages of these replications.


Scope

The activity of garlic will provide the way to fight against the growing hospital infection. Garlic will be used in the form of tablets in best concentration as a medicine against the pathogen of gastric intestinal tract i.e. Staph.


Resources

This study will be conducted by our group in the microbiology lab, at GPGCM, Abbottabad.


Result

According to objectives, 80% of Staph will sensitive to the concentration of 15 mg/ml of garlic extract.


Conclusion

We will conclude that in  the era of drug resistant bacteria, we need to focus on alternative drugs that have long history to avoid such emerging diseases and that could be easily available and affordable. In this study we will observe the concentration of the garlic extract and growth of test bacteria.

References

 

1.      Abiy E, Berhe A. Anti-Bacterial Effect of Garlic (Allium sativum) against Clinical Isolates of Staphylococcus aureus and Escherichia coli from Patients Attending Hawassa Referral Hospital, Ethiopia. J Infec Dis Treat. 2016, 2:2. doi:10.21767/2472-1093.100023

 

  1. Dikasso D (1999) Antiviral Effect of Garlic. Berhanena Selam Printing Press: Addis Ababa.
  2. Abebe D (2003) Medicinal plants. Berhanena Selam Press: Addis Ababa 17:1108-1112.
  3. Weiss RF (1988) Herbal Medicine. Hippocrates, VerlagStuttgart.
  4. WHO (1990) Medicinal Plants in Vietnam. WHO Regional Publications, Western Series, Manila.
  5. Abebe D, Ayehu A (1993) Medicinal plants and health practice in Ethiopia. Berhanena Selam Printing Press: Addis Ababa,pp: 219-221.
  6. Dikasso D, Tito TM(1999)Medicinal preparation and use of Garlic by traditional healers in southern Nations Nationalities and People’s State.Ethiop J Health Dev 13: 93-99.
  7. University of Mary Land Medical Center, Garlic Overview.

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