Methodology: The Antibacterial activity of the crude extract of garlic was investigated against Clinical and Standard isolates of S. aureus and E. coli by an Agar of both dilution and Cork borer techniques.Results and conclusions: The results showed that standard S.

aureus and E. coli were completely inhibited by 10 mg/ml and 15 mg/ml of agar media respectively and their clinical isolates were completely inhibited by 25 mg/ml, indicating that standard isolates are most sensitive and clinical isolates are least sensitive.Garlic could be used as effective antibacterial agent for these pathogenic microorganisms.Interests in plants with antimicrobial properties has come to use again because of emergence of resistance strains against antimicrobials such as penicillin [1].This study confirmed that the aqueous extract of Garlic had antibacterial effect against clinical isolates of S. aureus and E. coli.Even the forefather of antibiotic medicine Louis Pasture acknowledged garlic to be an effective antibiotic.Allicin is the antibacterial component found in Garlic.The researchers were able to study how garlic works at molecular level using allicin, garlic’s main biologically active component [7].Staphylococcus aureus is very important pathogen that causes a variety of diseases including skin infections.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.Plant materials.Garlic solution was obtained not from the whole part of the plant rather on the bulbs.Test organism.Two test organisms (S. aureus and E.

coli) which were clinically isolated from patients were collected from Awasssa Referal Hospital.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. aureus and Mackonkey for E. coli were prepared on a slant and on petri dishes [10].Antibacterial activity test.The antibacterial activity test of the crude extract of Garlic against both standard and clinical isolates were carried out by the Agardiffusion method [13].All the tests were carried out in triplicate and the results were reported as the averages of these replications.Table 2 showed that both clinical isolates of S. aureus and E.

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. aureus - + + + E. coli - - + + Standard S. aureus + + + + E. coli + + + +.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 1b: Result of antibacterial activity of Garlic extract compared with standard drugs against test organisms by Agar diffusion+indicates Inhibition-indicates growth.The result showed the both clinical and standard isolates of S.

aureus and E. coli were highly sensitive to concentrations of 0.75 ml/20 ml of agar media in using diffusion method and Cork borers.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 penicillin.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. aureus is known to be resistant.Then, I would like to appreciate the staff member of Ethiopian Health and Nutrition Research Institute/EHNRI/ especially the Drug Research Laboratory Heads Dr. Asfaw Debela, Dr. Dawit Dikasso and W/O Hirut Lemma for their valuable inputs to carry out the research at EHNRI.


A Comparison of the Antibacterial and Antifungal Activities of

DMTS, DETS, DATS and DPTS showed antibiotic activity against all bacteria in this study, resulting in clear inhibition zones, the size of which was dose-dependent (Fig.Figure 4 Plate inhibition zone assay showing the antimicrobial activity of different thiosulfinates against various bacteria and yeast.), Gram-positive bacteria (M. luteus) and yeast BY4742 cells.Therefore, considering antibacterial activity first, DMTS was most effective in this test system against P.

syringae 4612 and least effective against P. fluorescens whereas DATS was most effective against Micrococcus luteus and least effective against P.

fluorescens (Fig.All the thiosulfinates proved very effective against yeast BY4742 cells and resulted in relatively large inhibition zones in comparison to those for bacteria.To investigate the antibiotic activity of DMTS, DETS, DATS, DPTS and DBTS via the gas phase, a 20 µL drop of 80 mM test solution was placed in the centre of a Petri-dish lid and the Petri-dish base, containing medium seeded with bacteria, was inverted above the lid as previously described17.Thus, there was no contact between the test solution and the agar itself except by diffusion through the air.With the exception of DBTS, which was presumably not sufficiently volatile to achieve inhibitory concentrations, all thiosulfinates produced an inhibition zone above the droplet in the Petri plate lid for E. coli, P. syringae 4612 and M.

luteus.To test how much time was required for allicin to diffuse through the gas phase and achieve an inhibitory concentration at the seeded agar, a time-resolved experiment with allicin and E. coli was carried out.The results show that as little as one hour of exposure to allicin already leads to an effective growth inhibition of bacteria above the drop.Figure 6 Allicin (20 µL droplet of 80 mM solution) shows inhibitory effects against E. coli in this test dependent on exposure time, with maximum inhibition reached after only four hours exposure.Inhibition can be seen in comparison to growth on control plates without test substance.E. coli cells (Gram-negative) were hardly inhibited by 100 µM DMTS, DPTS or DBTS whereas allicin and DETS caused a high degree of inhibition after 36 h.

In the agar diffusion test, E. coli cells were inhibited by all test substances and this result demonstrates the importance of not relying on the conditions of a single test when assessing the antimicrobial effectivity of test compounds.Thus, the standard EUCAST procedure uses a low titre of cells in stationary culture, the agar diffusion test works with a concentration gradient, the drop test incorporates the substance at fixed concentrations and different cell densities are tested, whereas shake culture exposes the test cells under conditions of continuous agitation and high aeration.In the drop tests, Gram-negative P.

syringae 4612 cells were inhibited strongly by all thiosulfinates up to 48 h after plating out (Fig.Figure 7 Drop test illustrating the relative inhibitory activities of the thiosulfinates incorporated into growth medium at 100 µM on E. coli, P. syringae 4612 and M. luteus, respectively.Saccharomyces cerevisiae was used as a model fungus in drop tests on agar medium containing the test substance and in shake culture in 96 well plates (see next section) to assess the antimycotic activity of thiosulfinates.In the drop test, 10 µL of 10-fold serial dilutions were plated out onto control medium, or medium containing 5 µM of the test thiosulfinate.Thus, the mutants chosen are all appropriately relevant for testing and comparing the mechanism of action of allicin in relation to GSH metabolism, with respect to the other thiosulfinates (Fig.DMTS at 5 µM did not affect the growth of the wt or any of the mutant cells (Fig.DETS at 5 µM was not inhibitory for BY4742 wildtype, but clearly inhibited Δyap1, Δglr1, Δzwf1, Δgnd1 cells at this concentration.DBTS showed the highest toxicity to yeast of the thiosulfinates tested and again the drop test did not resolve any differential toxicity for DPTS at 5 µM between the wt and the mutants.Figure 9 Drop test of S. cerevisiae cells on CSM medium containing test thiosulfinates at 5 µM.The wt BY4742 was compared with the Δyap1 and other mutants.Figure 10 Effects of 50 µM thiosulfinates on the growth in shake culture of wt BY4742 and Δyap1, Δglr1, Δzwf1, Δgnd1, and Δtrx2 mutant yeast cells in CSM.(a) CSM alone (control); (b) DMTS; (c) DETS; (d) DATS (allicin); (e) DPTS.Figure 11 Effect of DBTS on the growth in CSM of wt BY4742 and Δyap1, Δglr1, Δzwf1, Δgnd1, and Δtrx2 mutant yeast cells in CSM and the synergistic effect of DMF with allicin.(f) 25 µM allicin and (g) 25 µM allicin in 0.5% DMF.Therefore, the effects of thiosulfinates on wt and Δyap1, Δglr1, Δzwf1, Δgnd1, and Δtrx2 yeast mutants were also investigated in shake culture because this has the additional advantage of providing relative growth kinetics and not just an end-point result48.In contrast to the stationary culture conditions in the MIC and MBC tests, or in drop tests where cells are plated onto medium containing the test substance, in shake culture cells tend to grow more robustly and generally tolerate higher concentrations of antibiotics.Previous experiments had shown that 50 µM allicin reduced the growth rate of wt BY4742 cells by approximately 50% at the end time point of 15 h in shake culture in CSM medium.Therefore, in these experiments we exposed the cells to 50 µM of thiosulfinates for comparison.DPTS at 50 µM was more inhibitory than allicin to the wt and again completely inhibitory to all of the mutants (Fig.This activity series can also be seen in the drop test, but the increased sensitivity of the mutants to DPTS compared to the wt was not resolved (Fig.DMF at this concentration showed no significant effect on growth of either the wt or the mutants in comparison to the CSM controls (Fig.However, a dose-dependent inhibition of wt and mutants can be seen at 5, 10 and 25 µM, with the latter concentration being completely inhibitory to the growth of all yeast strains (Fig.It can clearly be seen that the Δglr1 mutant is the most sensitive and this was completely inhibited for the duration of the experiment at 5 µM DBTS in 0.5% DMF.This very high degree of inhibition compared to the other thiosulfinates, suggests that DBTS had the greatest activity of the thiosulfinate series against Saccharomyces cerevisiae, a result which corresponds to the drop test results shown in Fig.Notably, a synergistic effect was observed between DMF and allicin, which at 25 µM showed a lesser inhibitory effect without DMF than with 0.5% DMF where it also completely inhibited the growth of wt and all mutant cells (Fig.To sum up the results of the chemogenetic profiling, the observation that the chosen mutants were generally more sensitive to thiosulfinates than the wt suggests that the other thiosulfinates are probably acting similarly to allicin and targeting the cellular GSH pool and GSH metabolism as well as resulting in protein thiol oxidation3,20,36,38.Thus, it seems that GSH is the first line of cellular defence and the ability of the cells to reduce GSSG to GSH is crucial for the cells’s resistance to allicin and the other thiosulfinates.Nevertheless, the tendency that DMTS was least toxic and allicin most toxic to A549 cells was a clearly visible trend in all the experiments and the data confirm that the analogues are of similar toxicity to allicin.The concentration-dependent inhibition of root growth by thiosulfinates is shown in Fig.The enhanced sensitivity of GSH metabolism mutants compared to the Col-0 wt confirms the results of the chemogenetic screen with yeast mutants (Figs. .

Effect of garlic on bacterial biofilm formation on orthodontic wire

Despite its antibacterial function, garlic extract increases biofilm formation by S mutans to orthodontic wire, likely through upregulation of glucosyltransferase expression.Growth inhibition of oral bacteria was tested after 50 µL of garlic extract was placed on an agar plate.After cultivating streptococci in biofilm medium (BM)-sucrose with garlic extract and orthodontic wire, adenosine triphosphate (ATP) measurement and viable cell counting was performed from the bacteria attached on the wire.8 In the present study, adenosine triphosphate (ATP) assay, viable cell counting, and scanning electron microscopic (SEM) analysis were performed on in vitro growth to investigate the effect of garlic on microbial attachment to orthodontic wire.Since S mutans exists almost exclusively in oral biofilms and is considered the primary etiologic agent of human dental caries, 7 we evaluated the effect of garlic extract on biofilm formation by S mutans on orthodontic wire in vitro.After the inoculum had dried, an 8-mm filter paper disk impregnated with 50 µL of garlic extract was placed onto an agar plate and incubated overnight at 37°C in aerobic condition.After 40-hour incubation at 37°C under aerobic condition, each wire was washed twice in sterile PBS (pH 7.2) and moved to another sterile Eppendorf tube.The average luminescence value of negative control wells containing PBS buffer and ATP assay solution was subtracted from each luminescence value.The relative expression levels of GTF family genes were normalized to those of 16S rRNA in the same samples.ATP assay to measure bacterial cell growth on orthodontic wires treated with garlic extract.ATP assay to measure bacterial cell growth on orthodontic wires treated with garlic extract.Measuring the effect of garlic on bacterial attachment to orthodontic wire, we interestingly found that the relative luminescence of wire-attached S mutans and S sobrinus ( Figure 1a ) increased continuously in a concentration-dependent manner in both bacteria, suggesting more bacterial cells attached to the orthodontic wire in the presence of garlic extract (S mutans, P = .01 for 16 mg/mL garlic extract; S sobrinus, P = .006 for 16 mg/mL garlic extract).MICs against S mutans and S sobrinus were 32 mg/mL and 64 mg/mL, respectively.Garlic extract has a wide spectrum of antibacterial activity, affecting Escherichia, Salmonella, Staphylococcus, Streptococcus, Klebsiella, Proteus, Clostridium, Mycobacterium, and Helicobacter species.13,14 Previous reports have shown a synergistic antibacterial effect when garlic extract and antibiotics are combined.15 Some oral streptococci have been shown to be sensitive to garlic extract, and a mouthwash containing garlic extract effectively reduced the total salivary bacterial and mutans streptococci counts.16 Shuford et al.17 demonstrated that fresh garlic extract inhibited growth of Candida albicans in its planktonic, adherent, and sessile phases, raising the question of whether garlic has an antifungal effect on Candida albicans biofilm, and if so, what the underlying inhibitory mechanism is.In this study, we tried to uncover the effect of garlic extract on dental biofilm formation using S mutans by analyzing attachment on orthodontic wire following garlic extract treatment.The GTFs, in concert with glucan-binding proteins, contribute greatly to initial attachment and to the formation of biofilms.18 A previous study showed that sub-MICs of allicin may play a role in the prevention of adherence of Staphylococcus epidermidis to microtiter plates.19 ,S epidermidis biofilm formation is known to be associated with the production of the polysaccharide intercellular adhesin (PIA), poly-N-acetylglucosamine polysaccharide (PNAG), and recent evidence indicates that staphylococcal accessory regulator (SarA), a central regulatory element that controls the production of S aureus virulence factors, is essential for the synthesis of PIA/PNAG and ensuing biofilm development in this species.20 These results suggest that the enzymes participating in bacterial biofilm formation are specific to bacteria, and that the increase of bacterial attachment by garlic extract through upregulation of GTF family genes expression is therefore very specific to S mutans.Therefore, some components of garlic extract in our study could have induced effective bacterial biofilm formation to wire.Another possibility is that the increase in biofilm formation by garlic extract was caused by pH change of the medium.We thus concluded that the increase in bacterial attachment on wire due to upregulated GTF expression was induced by garlic extract itself. .

Inhibitory effect of garlic extract on oral bacteria

The garlic extract (57.1% (w/v), containing 220 μg/ml allicin) inhibited the growth and killed most of the organisms tested. .


Therefore, to know the effect of food additives on survival of probiotic is important.Disk diffusion assays revealed that while Bifidobacterium longum BB536 is the most susceptible (49.37 ± 1.07 mm), Lactobacillus acidophilus 74-2 is the most resistant (no inhibition zone) bacteria to the fresh garlic juice, significantly.Evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. ).Journal of Probiotics and Health, 1(101), 1-11. phase is a distinct growth phase that prepares bacteria for exponential growth and involves transient metal accumulation.

).The growth rate and the doubling time are obtained during this phase. ). ).

).Food Science & Nutrition, 3(1), 48-55. of garlic and dill extract on yoghurt probiotic bacteria (Bifidobacterium bifidum and Lactobacillus acidophilus) and their role in rats triglycerides and cholesterol.and Lactobacillus acidophilus. ).The objective of this research is to investigate the growth curves of yoghurt starter cultures (Lactobacillus bulgaricus and Streptococcus thermophilus) and three commercial probiotic bacteria (Lactobacillus acidophilus 74-2, Lactobacillus rhamnosus HN001TM, and Bifidobacterium longum BB536).Hence, the second aim is to determine the effect of the garlic on selected starter culture and probiotic bacteria strains.For only B. longum, a MRS broth was sterilized at 118 °C for 15 minutes as instructed by the supplier, and the broth was sterilized at 121 °C for 15 minutes for other bacteria strains.

).To determine the growth curve of B. longum, a bacterial culture was added to the MRS broth, and then 24 individual tubes were obtained.Journal of Food Research, 2(1), 158. .

).2.5 Effect of garlic on test strains. ).In another experiment, each bacteria strain was inoculated in the 100 mL MRS broths with 0.02% GG and control medium, then were enumerated at the 0 and 24th h. Lactobacillus bulgaricus, Lactobacillus rhamnosus, and Bifidobacterium longum were incubated under anaerobic conditions, while others were incubated under aerobic conditions as indicated in Table 1.The relationship of bacterial growth and pH was evaluated using correlation analysis. ).Figure 1 illustrates the growth curves of the bacteria and the pH change in the medium during incubation.Since there was no significant difference between triplicates for each bacteria strain, the growth curve data were obtained by taking mean values into account.Growth curves of test strains and pH values in MRS broth at 37 °C.For L. bulgaricus, it was found that the first 2 h were the lag phase, and in the next 3 h microorganisms exhibited rapid growth, which was called the exponential phase (Figure 1A).After 24 h of incubation, the decrease in pH was determined as 2.35 units, which reached a more acidic pH than others, and the correlation between growth and pH was determined as moderately negative.Journal of Applied Pharmaceutical Science, 4(11), 8-14. determined that, after the first 2 h, L.

bulgaricus entered the exponential phase as in our study, and the specific growth rate was found to be 0.47 h−1 in SPY-10 (Soy Peptone Yeast) medium at 37 °C.reported that the specific growth rate of four different strains of S. thermophilus, which were activated in the M17 broth at 43 °C, ranged between 0.610-0.674 h−1.Similarly, the specific growth rate obtained in this study was 0, 619 h−1.The effect of temperature and pH on the growth of lactic acid bacteria.a pH-auxostat study.International Journal of Food Microbiology, 85(1-2), 171-183. , the specific growth rate of S. thermophilus St20 was defined 2.2 h−1 at 45 °C in modified medium.International Journal of Food Microbiology, 129(3), 211-220.

). ). ).It is known that at the beginning of the incubation, S.

thermophilus grow faster than L. bulgaricus.The MRS broth medium decreased from 5.40 to 3.91 pH at the end of 24 h, and the correlation between pH and growth between pH and growth was not found statistically significant.Studies on probiotics properties of two lactobacillus strains. stated that L. acidophilus’ (B / 103-1-5) growth curve was obtained by optical density measurement at 2 h intervals at 37 °C for 24 hours in a MRS liquid medium, the specific growth rate was 0.493 h−1, and the doubling time was 1.4 h. The shorter doubling time of L. acidophilus 74-2 used in the study shows that the values are different in a strain-by-strain comparison despite having the same temperature and medium.

, L. acidophilus strains were found to be similar in terms of growth parameters under the same growth conditions.As shown in Figure 1D, with L. rhamnosus HN001TM, the lag phase continued for the first 3 h after the exponential phase was observed, between 3 and 4 h. The specific growth rate was 1.93 h−1, and the doubling time was 0.36 h.

MedveDova´ et al. (2008)MedveDova´, A., Lipt’akov’a, D., & Val’ık, L. (2008).The results revealed that the influence of different temperature and media on growth was significant, and the doubling time decreased when the temperature increased.

, the specific growth rate in the MRS broth medium at 37 °C was 0.82 h−1. ).In the current study, the growth patterns of L.

acidophilus and L. rhamnosus are similar in terms of statistics under the same growth conditions.Compared to yoghurt bacteria and other probiotics, it was determined that B. longum was the bacteria that minimally changes the pH value in the MRS liquid medium which was statistically significant (Table 2).

). , studying the growth kinetics of Bifidobacterium adolescentis MB239 strain with different carbohydrate sources, found that the maximum biomass yield and specific growth rate were at pH 5.50, but the acid production did not have linear correlation with these parameters; the lowest level was found to be pH Effect of garlic on probiotics and yoghurt bacteria.The enumeration of each bacteria in the control and the broth containing 0.02% GG revealed that the probiotic and yoghurt bacteria have not been adversely affected in a significant way by the given ratio of garlic.The effect of garlic on yoghurt and probiotic strains.In vitro effects of food extracts on selected probiotic and pathogenic bacteria.International Journal of Food Sciences and Nutrition, 60(8), 717-727. ).Probiotics and Antimicrobial Proteins, 6(2), 82-87. Lactobacillus acidophilus.African Journal of Microbiological Research, 7(8), 669-677. where L.

acidophilus La14 150B did not show an inhibition zone against a 30 µL garlic extract. ).Inhibition effects against some pathogen food-borne bacteria of lactic acid bacteria and some spices (Master’s thesis).It was observed that B. longum BB536 was the most sensitive bacteria, and the susceptibility pattern was S. thermophilus, L. rhamnosus HN001TM, and L. bulgaricus with 28.16 ± 1.17, 18.81 ± 0.82, and 15.11 ± 1.49 mm zone diameter, respectively (Table 3).and Lactobacillus acidophilus.African Journal of Microbiological Research, 7(8), 669-677.

determined that Bifidobacterium lactis Bi-07 300B was the most resistant, and B. longum BB536 was the most susceptible bacteria in the study on the effect of garlic among Bifidobacterium spp.This result proves that different garlic origin or extraction method may have different antimicrobial effects on the same strain.As far as we know, this is the first study researching the effect of garlic on yoghurt bacteria by disk diffusion assay although there are numerous studies examining the inhibitory effect of garlic on some pathogenic microorganisms.In this study, it was determined that both bacteria were adversely affected, and S. thermophilus was more sensitive than L. bulgaricus against FGJ, but a 0.02% GG aqueous solution was not effective enough to inhibit growth.Therefore, the growth characteristics of certain strains and the effect of food additives on the strains before usage in a new food matrix is important knowledge.


5 Natural Antibiotics to Try at Home

Certain plant extracts, essential oils, and even foods have antibiotic properties.A small sampling study of 58 Chinese plants found that 23 had antibacterial properties and 15 had antifungal properties.To use honey as an antibiotic, apply it directly to the wound or infected area.The honey can help kill off the bacteria and aid in the healing process.A 2011 study found that garlic concentrate is effective against bacteria.You can purchase garlic concentrate or extract at your local health food store.If you’re taking a garlic supplement, be sure to follow the dosage directions as provided.Option 5: Oregano essential oil Share on Pinterest Carvacrol is an ingredient found in oregano essential oil.You shouldn’t ingest oregano essential oil or use undiluted essential oil on the skin.lemon Buy oregano essential oil here. .

Health Effects of Garlic

Sulfur compounds, including allicin, appear to be the active components in the root bulb of the garlic plant.C 5 to13 Patients at risk of thrombosis should be advised that garlic may have a modest but significant effect on platelet aggregation compared with placebo.C 5 to13 Patients at risk of thrombosis should be advised that garlic may have a modest but significant effect on platelet aggregation compared with placebo.Historically, garlic has been used around the world to treat many conditions, including hypertension, infections, and snakebites, and some cultures have used it to ward off evil spirits.Allicin is formed when alliin, a sulfur-containing amino acid, comes into contact with the enzyme alliinase when raw garlic is chopped, crushed, or chewed.The antimicrobial, hypolipidemic, antioxidant, and antithrombotic effects that have been attributed to garlic are thought to be related to allicin and other breakdown products.Uses and Efficacy Garlic has been studied extensively in vitro, in animal and human clinical trials, and in epidemiologic evaluations for its multiple medicinal properties.A European trial12 comparing garlic with a commercial lipid-lowering drug (bezafibrate, a fibric acid derivative not available in the United States) found them to be equally effective in decreasing lipids to a statistically significant extent.Only three trials showed a statistically significant reduction in diastolic blood pressure (2 to 7 percent), and one showed a statistically significant reduction in systolic blood pressure (approximately 3 percent) in patients treated with garlic compared with placebo.Contraindications, Adverse Effects, Interactions The ingestion of one to two cloves of raw garlic per day is considered safe in adults. .

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