Doxycycline and amoxicillin

This research provides scientific bases for the notion that allicin is a broad-spectrum antimicrobial, capable of warding off different types of infections. It is likely that bacteria would develop resistance to allicin because this would require modifying the very enzymes that make their activity possible scientists found that allicin blocks the enzymes by reacting with one of their important components known as self hydyl (SH) groups, or

thiols

Staphylococcus aureus is very important pathogen that causes a variety of diseases including skin infections.

Gastro intestinal disease (*staph*) food poisoning, toxic shock syndrome and nosocomial infections acquired during hospitalization.

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

Escherichia coli is a bacteria that exists naturally as part of normal gut of healthy humans and mammals.

However, there are relatively few strains of this strains pf this organism, which are pathogenic to humans and are associated with

food

aureus that are resistant to penicillin first appeared in clinical specimens in early 1950s. Soon thereafter, multiple antibiotic resistance was detected in chemical isolates of S. aureus ; these strains was resistant to macrolide antibiotic, amino glycoside, and tetracycline. Plasmids and transposons are clearly important in conferring and transferring antibiotic resistance between bacteria.

Although plasmids and transposons are certainly involved, the actual evolutionary mechanisms underlying this phenomenon have yet to be explained.

One this consequence is the emergence of epidemics hospitals strains of S.

aureus that is resistant to virtually all useful antibiotics, including methicillin and vancomycin. These strains are currently a significance cause of nosocomial (hospital acquired) infections in parts of the world [11]. Ethical clearance was obtained from Ethical board of Debub University and support letter were delivered to Ethiopian Public Health Institute and Hawassa Referral Hospital. Garlic solution was obtained not from the whole part of the plant rather on the bulbs.

The bulb of Garlic were peeled, weighted and then ligated using pestle while adding small amount of H 2 O. The extracts then allowed to freeze at -18°C (deep freezing) so as to concentrate the chemical (allicin). Then after freezing, the filtrate was put into lyophilizer till an amorphous powder weighted and then diluted with distilled water and used for the experiment [12].

coli ) which were clinically isolated from patients were collected from Awasssa Referal Hospital.

Two standard pure cultures were collected from EHNRI (Ethiopian Health and Nutrition Research Institute). Both clinical and standard were sub cultured into subsequent Nutrient broth and definite Medias in which they favored i.e., Mannitol salt agar (MSA) for S. coli were prepared on a slant and on petri dishes [10]. Cultured of the test organisms were maintained on nutrient both.

Briefly, four to six colonies were picked with an inoculating loop and suspended in 5 ml of broth and incubated at 37°C for 24 hours. The turbidity of the broth culture was then equilibrated to match that of 0.5 Macfarlands standards.

This provides organisms in the range of 1 ? 10 6 to 5 ? 10 8 cfu/mol which is pathogenic that used for the test [12].

The antibacterial activity test of the crude extract of Garlic against both standard and clinical isolates were carried out by the Agardiffusion method [13].

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.

A separate agar plate without sample or drugs was also prepared in order to provide an appropriate growth of organisms.

(As the same time as control) Two standard drugs as a positive control were also tested against these microorganisms. These were 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 was determined by direct visual comparison of the growth of the test cultures.

All the tests were carried out in triplicate and the results were reported as the averages of these replications.

In this methods 0.2 ml of garlic extract was mixed with 20 ml of sterile nutrient agar using a mixer (vortex), and then poured into sterile Petri dishes.

After congealing, the seeded agar was punched out with a sterile bore (back hole of 10 ml pipette diameter=9 mm) at equally spaced out positions to make four holes.

Four of the holes were filled with 0.1 ml of the test sample solution while

the

The antibacterial activity was evaluated by measuring the diameter of the zone of inhibition using ruler (the media in both methods was prepared according to the instructions that the manufacturer orders (written in the flask that for how many gm of powder how money ml of distilled water is enough).

The results of the susceptibility of the test organisms against the garlic extracts which are stated below in Tables 1a and 1b . coli were sensitive to the concentration of 15 mg/ml (0.75 ml/20 ml of agar media) which is about 80% but about 10% of the organisms were not sensitive for lower concentrations i.e., for 0.25 ml in media.

Isolates Micro organisms Concentration of Garlic/20 ml 0.25 ml 0.5 ml 0.75 ml 1.5 ml Clinical S. coli + + + + Table 1a: Result of Antibacterial activity of Garlic aqueous extract by agar diffusion method. Antibiotic type Concentration Size of Clear zone Organisms presence or absence S.

aureus E.coli Garlic 5mg/ml Small - - 10mg/ml Small + - 15mg/ml Medium + + 25mg/ml Large + + Tetracycline 30mg/ml Very small - + Chloramphenicol 10mg/ml Large + + Penicillin 30mg/ml Medium - +

Table

Comparatively, Garlic extracts inhibited bacterial growths than Tetracycline and Penicillin.

The result showed the both clinical and standard isolates of S. coli were highly sensitive to concentrations of 0.75 ml/20 ml of agar media in using diffusion method

and

Therefore, the garlic extract was more important for the prevention of resistant S.

aureus which is currently becoming a challenge developing resistance to many commercially available drugs

like

In this study we have observed that, as the concentration of the garlic extract increases we have seen efficiency increased and hence inhibition and growth of test bacteria has been diminished. As observed from the above tables, Larger clear zones at higher concentrations and lower clear zones at lower concentrations.

This implies that, Garlic has both bacteriostatic and bactericidal effect. Based on the results of this study which showed garlic to make large clear zones than currently available antibiotics used in the study Garlic could be used as an effective antibacterial agent in Ethiopia where S. It could be made as a tablet in the best concentrations and affordable dosages so that it can be used as medicine to these two pathogenic gastro intestinal enteric.

In the era of those 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.

One can use garlic as a member of the daily diet for better health especially the fresh garlic.

This research can be used as a base to well identify the actual allicin minimum inhibition concentration. The authors declare that they have no any competing interests.

EA and AB designed the study methodologies and EA carried out the investigation and AB edited the manuscript. Materials used for the study were stored in Hawassa University, Department of Biology Laboratory.

But the files are fully presented in this manuscript.

First of all, I would like to thank my God who gave me the strength and patience to do the research.

Then, I would like to appreciate the staff member of Ethiopian Health and Nutrition Research Institute/EHNRI/ especially the Drug Research Laboratory Heads Dr. Dawit Dikasso and W/O Hirut Lemma for their valuable inputs to carry out the research at EHNRI. Next, I would like to thank my advisor Ato Asefaw Berhe who facilitated ways to work on this project and shared me important ideas for the research.