Invitro Determination Of Bacteriocidal Effect Of Garlic On Staphylococcus Aureus

Project and Seminar Material for Microbiology

Invitro Determination Of Bacteriocidal Effect Of Garlic On Staphylococcus Aureus


Invitro determination of bacteriocidal effect of garlic extract on staphylococcus aureus causing skin and urinary tract infection (UTI) on some female patients were studied. High vagina swab (HVS) samples were collected randomly from selected number of patients who had symptoms similar to that of Staphylococcus aureus activities. The samples were inoculated on Macconkey agar and nutrient agar and incubated at 370c for 24-48 hrs. Colonies that appeared pinkish in whitish creamy and yellowish on nutrient agar were gram stained and examined microscopically. Also, biochemical tests were carried out in order to characterize the organism present in each positive sample 80% of the samples examined were found to react positively to Staphylococcal infections. It’s prevalence is more among the age range of 18 – 30 years. It is suggested that government should encourage public health education on the potency of the cheap and readily available antibacterial agent like garlic and it’s addition to daily dietary which will act as a prop drug especially to women to reduce the incidence of S. Aureus that causes skin and urinary tract injection. Garlic extract has been proved to be an excellent cure to the ailment.

Table of Contents

Chapter One

1.0 Introduction

  • 1.1 Aim and objectives
  • 1.2 Hypothesis
  • 1.3 Delimitation
  • 1.4 Statement of problem

Chapter Two

2.0 Literature Review

Chapter Three

3.0 Materials and Method

  • 3.1 Bacteriological culture
  • 3.2 Sterilization
  • 3.3 Collection and transport of specimens
  • 3.4 Laboratory examination of H.V.S. Sample
  • 3.5 Biocehmical characterization of isolates catalase
  • 3.6 Procurement and extraction of garlic extract
  • 3.7 Serial dilution
  • 3.8 Disc preparation
  • 3.9 Impregnation of thegarlic extract into the paper disc.
  • 3.9.1 Sensitivity testing of garlic extract

Chapter Four

4.0 Result

  • 4.1 Preliminary identification of bacteria isolate
  • 4.2 Biochemical test showing staphyloccoccus aureus
  • 4.3 Sensitivity testing using garlic extract and three antibiotics indicating their zone of inhibition.

Chapter Five

5.0 Discussion, conclusion and recommendations

  • 5.1 Discussion
  • 5.2 Conclusion
  • 5.3 Recommendation
  • References

List of Tables

  • The level of bacteria according to age range
  • Preliminary identification of bacteria isolate
  • Biochemical test carried out to identify staphylococcus aureus
  • Sensitivity testing using garlic extract and three antibiotics
  • Table for minimum inhibitory concentration
  • Bacteriocidal evaluation of garlic extract

List of Figures

  • Chemical Structure of Allicin
  • Flow chart of garlic extract
  • Flow chart of serial dilution
  • Disc before incubation
  • Disc after incubation

Chapter One

1.0 Introduction

Allium satirum commonly known as garlic is an odoriferous plant belonging to a family lilacease. It is a small perennial herb with narrow flat leaves and surrounded by membranous patches, it is grown mostly in Northern Nigeria. It is used primarily for seasoning or spice and also for its medicinal property. The plant contain a medicinal properly just like other plants such as aloes, Indian liquorices, alligator pepper etc which are also found useful, although the extend is not accurately recorded. The medicinal property of garlic is due to its “sulphur” content which was believed to be responsible for it’s medicinal value. Garlic as a medicinal plant has been widely used and found to be effective on number of serious infections. Other plants like the ones mentioned earlier also have their medicinal properly, some which is as a result of the presence of ackloids, volatile oils, polyphenol and some related sulphur compound contained in them. Similarly, some are found to be used as vermifuge, stimulating carminative toxic and also as condiments and for treatment of worm bites just like garlic.

The diluted juice of the fresh bulb is used externally as a mild antiseptic and the finely chopped cloves added to food act as an effective vermifuge. By the distillation of the bulb, a volatile oil consisting mainly of daily disulphide and allyl – prop disulphide is obtained. It is used to diminish cough in chronic bronchitic. Meanwhile, in this project work, we shall be working on garlic’s bacteriocidal effect on staphylococcus aureus. Garlic is one of the best known natural plant that provide sulphur. Many of the sulphur containing compounds in garlic such as dietlyl – disulphide, propienyl methyl sulphide, allicin which stands as organic disulphide present in garlic inactivates the amino acid. Cysteine and such reactions of the garlic disulphide present in garlic may inhibit protein synthesis in microorganism. This reaction may have contributed to the bacteriocidal effect of garlic on bactenal cell. Garlic also contain a glycoside, vitamin B,C and D and allisation I and II, the volatile sulphur oil which form part of a vermifugal medicinal property. An antibiotic substance “allicin” which appear to be more bacteriostatic than bactericidal has been isolated from the cloves. Barley (1950). It has antibiotic effect on both gram positive and gram negative bacteria. Example Salmonella typhinurium, staphylococcus aureus, Bacillus Species, Eshericha Coli, etc. (Shelef, 1980).

Staphycoccus aureus is a predominant causative agent of skin and urinary tract infection among men and women. Therefore, urinary tract infection is referred to as infection affecting the urinary system comprising invasion of any of the tissues that make up the tract, extending from the renal cortex to the urethral meatus. Kunin, (1979). These diseases include cystitis – inflammation of bladder, Urethiritis – information of the urethra. Also, skin infection is referred to as infection affecting the skin. The organisms are present on the whole skin or part of the skin surface including those in deeper parts of the skin such as the ducts of sebaceous and sweat gland follicles.

Staphylococcus aureus is a gram positive bacteria, coagulase-positive and a human pathogen which live as a commensal in the anterior nares of over half the normal population. It may also colonize the skin. It is the most common cause of acute pyogenic (pus-producing)) infection in man. It has a marked predilection for the skin and surface structures and causes boils, carbunches, pustules, styles, impetigo (often in association with streptococcus pyogenes), pemphigus neonatorum, and sticky eyes in babies. Acute osteomyelitis in children and mastitis in lactating mothers. It is the commonest cause of infection of wounds and bums. Other staphylococcal species are coagulase-negative (Jackson, M.H. CRAWFORD, J.F, MORIS, G. and SMITH P.G. 1989)

1.1 Aim and Objectives

  1. To assess the bacteriocidal property of garlic as alternative to synthetic antibiotics
  2. To know the concentration of garlic extract that will be effective on the pathogen.

1.2 Hypothesis

  • HO:- Garlic extract has an inhibitory effect on Staphylococcus aureus isolated from urinary tract infection (UTI) among women.
  • HIL:- Garlic extract does not have an inhibitory effect on Staphylococcus aureus isolated from urinary tract infection (UTI)
  • HII: High concentration of garlic extract is needed an inhibitory effect on Staphylococcus aureus UTI among women.

1.3 Delimitations

  1. This project work will be limited on only Staphylococcus aureus causing urinary tract infection among women
  2. Due to time and financial factors, sample and materials collection were limited to Enugu metropolis.
  3. It is also limited to only the efficacy of garlic extract and the concentration that yield the highest efficacy.
  4. This work did not go further to determine the normal dose (potency) that will be taken in order to avoid adverse effect in hypersensitivity reactions.

1.4 Statement of Problem

  1. Garlic extract (drug) lose their efficacy after a period of time.
  2. Contamination of drug by opportunistic micro organisms during processing.

Chapter Five

5.0 Discussion and Conclusion

5.1 Discussion

Staphylococcus aurues which was found to e sensitive to garlic extract (GE) has a mean zone of inhibition of various potencies from 30mg to 50mg which is equipmetn to urinary tract infection (UTI) antibiotics like Nalixidic acid, and Gentamycin as broad specstrum as shown in table IV, this is in agreement to Shelef,(1980). Example, 30mg of garlic extract (GE) with inhibition zone of 13mm is equivalent to 30mg of Nalididic drug inhibition zone of 13.5mm. The highest mean h was compared with control drug, although the potencies are not the same is 50mg of high concentration garlic extract (GE).

Crude extract which were found not active against the test organism as seen in table VI may be due to the fact that were singly and therefore may need to be mixed with other extracts of active ingredients to be active. The sensitivity of micro organisms to plant coponets are advanced from the world of antibiotics when used against other micro organism can check the proliferation of microbial infection and are useful for their control staphylococcus aureus culture rate which was discovered to be more prevalent in the age range of 18-30 years, could be attributed to unhygienic measure and casual sex due to their sexual activeness in agreement Kunnin, (1979). While there was low prevalence at the age range of 61-70 years. Therefore, Staphylococcus aureus causing infection of the skin and urinary tract infection (UTI) as shown by the result is sensitive to garlic extract at high concentration, which implies that the more concentration of extract, the higher the magnified zone of inhibition to be seen, which is also equivalent to more intake of garlic extract (GE), the more Staph aureus is to be sensitive to it.

5.2 Conclusion

From the sensitivity testing, Garlic extract which was found to be effective on Staph aureus causing skin and urinary tract infection among women have no record on the effect of too much intake except on much exposure to garlic dust based on commercial extraction level. Then if one subject herself to garlic intake, there is every tendency of getting cured of staph aureus causing skin and urinary tract infections. To determine the max, mum potency that will be fixed as regards to other antibiotics regarded as antibacterial agent is the task ahead

5.3 Recommendation

Antibiotics like Nalidioxic acid, Nitrofurantoin and Genmntamycin were said to be effective against Staph aureus causing skin and urinary tract infection but the drugs are costly. This work has proud that high concentration of garlic extract is effective on staph aureus causing urinary tract infection and skin infections therefore, we recommended that:-

Further work be carried out to determine the normal dose that will be taken so that when one takes it, he/she will not experience adverse effect or hyperactivity. Many people mostly women have been staking garlic as spice without the knowledge of its medicinal potency, in light of this we recommend that the governments should encourage public health /education on the potency of the cheap and readily available antibacterial agent like garlic and its addition to daily dietary will act as prophylactic drug especially to women in the case of Staphylococcus aureus causing urinary tract infection and other skin diseases.

In order that a traditional folk could be fully accepted large scale controlled clinical trials must be enclosed and conducted by food drug Administration.

In the absence of an altruistic pharmaceutical firms steeping forward, the government or a public oriented organization might take the initiative to test garlic and other such agents for their therapeutic claims. With this is will encourage the garlic champions of present to come forward to prove the medicinal worth of this pungent plant.
This research work was limited to Staph aureus¬ alone and other research world should be carried out in other related bacterial like E. Co, Bacillus species and salmonella typhyrium and others.

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