Antibacterial and Antivirulence Effects of Kale (Brassica oleracea var. sabellica) on Streptococcus intermedius
Kittipong LaosuwanChaiyakorn SongsangwattanakijKathawut TachasuttirutSurawut PongsiriwetJitjiroj IttichaicharoenSunee Chansakaow
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Persistent odontogenic infections caused by resistant bacterial species, such as , have consistently been associated with deep-seated infections. This in vitro study aimed to evaluate the antibacterial and antivirulence effects of kale () on . was freshly incubated in tryptic soy broth media. Three experiments per concentration of kale were conducted under aseptic conditions (i.e., disc diffusion, broth microdilution, and reverse-transcription polymerase chain reaction) to evaluate the antibacterial and antivirulence effects. The samples were then treated with 1000, 500, 250, 125, 65, 30, 15, 7, and 3 mg/mL kale; ampicillin (positive control); and tryptic soy broth (negative control). After 24-h incubation, the inhibition zone, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and relative gene expression of the virulence factor (intermedilysin []) were measured. All assays were conducted in triplicate. The findings were reported and analyzed as means ± standard deviations. The agar disc diffusion and relative gene expression were statistically analyzed using one-way analysis of variance and Tukey’s test, with the significance level set at P < 0.05. Kale showed antibacterial effects on by significantly inhibiting bacterial growth and reducing expression only at a concentration of 1000 mg/mL; it yielded an inhibition zone of 11.12 ± 1.59 mm, which was smaller than that with ampicillin. The MIC and MBC ranged from 15 to 65 mg/mL and from 500 mg/mL, respectively. Conversely, the highest concentration of kale yielded significantly less inhibition than did ampicillin. The antibacterial effects of kale may be dose-dependent. Kale can inhibit bacterial growth and suppress expression under in vitro conditions of , which is mainly involved in deep-seated odontogenic infections.Keywords:
Minimum bactericidal concentration
Tryptic soy broth
Agar diffusion test
Plants with medicinal value produce certain chemical elements known as phytochemicals that have antibacterial activity. The study was aimed at determining the antibacterial activity of Vernonia amygdalina against bacterial isolates using agar well diffusion method. In addition, the phytochemicals analysis of the extracts was also determined. The phytochemical analysis showed the presence of saponins, steroids, terpenoids, tannins, alkaloids, and flavonoids. The result of Vernonia amygdalina showed that the average zones of inhibitions observed against these bacterial ranges from 6-22mm. The highest zone is also exhibited against E. coli with average diameter of zone of inhibition of 22mm. At 100mg/ml concentration for Samonella, the zone of inhibition was recorded to be 21mm while at 12.5mg/ml there was no inhibition. At 25mg/ml and 12.5mg/ml, against Pseudomonas there was no inhibition. In other to further confirm the activity of these plant extracts, the minimum inhibitory concentration and minimum bactericidal concentration was determined and the result showed that the extract exerted good antibacterial activity on all the test organisms at different concentration. The result of minimum inhibitory concentration ranges from 10 to 12.5mg/ml and that of MBC ranges from 5 to 20mg/ml. It is worthy to note that MBC values is greater than that of minimum inhibitory concentration. The study provides insight into the antibacterial activities of the plant extracts and its use in the treatment of bacterial infections.
Vernonia amygdalina
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Minimum bactericidal concentration
Agar diffusion test
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This study aimed at evaluating the health benefits of popular Moringa oleifera leaf. The aqueous and methanolic extracts of the leaf at two different concentrations (1:1 and 1:2) was used to determine the phytochemical screening and its antibacterial activity. Escherichia coli, Streptococcus pneumonia and Staphlococcus aureus were used in this study, applying agar diffusion methods. The phytochemical screening indicated presence of secondary metabolites such as alkaloid, flavonoids, anthraquinoines, tannins and phenol in both extracts making it to have antibacterial potentials. Both extract showed remarkable activity against the growth of the selected bacteria; nevertheless, the methanol extract had more antibacterial activity than the water extract, more so the extracts were discovered to be more active at higher concentration. The water extract was not active at low concentration, that is 1:1 but had diameter zone of inhibition of 10 mm each for 1:2 concentration. The minimum inhibition concentration (MIC) that inhibits these bacterial ranged between 1:4 and 1:16 and the minimum bactericidal concentration (MBC) that kills the growth of the bacterial isolates completely was 1:16. The result of this study showed that M. oleifera could be a valuable antibacterial drug in the treatment of infections caused by the test organisms. Key words: Agar diffusion method, aqueous and methanol extracts, secondary metabolites, zone of inhibition, minimum inhibition concentration (MIC), minimum bactericidal concentration (MBC).
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Moringa
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Bacterial growth
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Since antibiotic resistance of bacteria is becoming such a large and frequent problem, this research was conducted to determine if resistance in Escherichia coli could be induced by repeated exposure to ampicillin.The procedure was based on a standardized method of testing resistance called the Kirby-Bauer (KB) Disk Diffusion Test. Brain-Heart Infusion (BHI) broth was labeled CØ for control and Tl-5 for tests. The broths were inoculated with E. coli. Ampicillin disks were added to the Test Group's BHI broths. After incubation, the BHI broths were sub-cultured to blood agar plates and Mueller-Hinton agar plates for the KB test. The plates were incubated, and the zones of inhibition read. The E. coli was then sub-cultured from the blood agar to the BHI, and the process was repeated until the E. coli showed significant resistance to the ampicillin.The average rate of resistance of the CØ Group was 0 mm per day, because the zone diameters of the CØ Group stayed at 20 mm. The average rate of resistance of the Test Group was 3.2 mm per day. This data indicated that the CØ Group remained susceptible to the ampicillin throughout the experiment. The Test Group, however, did become resistant to the ampicillin.
Brain heart infusion
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Agar diffusion test
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The use of bacteria in the synthesis of silver nanoparticles (AgNPs) emerges as an ecofriendly and exciting approach. In the present study, we reported the biosynthesis of AgNPs by using culture supernatant of the bacteria Bacillus licheniformis (MN900686). The biogenically synthesized AgNPs were confirmed by the change in the color of the culture filtrate from yellow to brown after the addition of AgNO3. Further characterization performed by means of UV vis-spectroscopy showed absorption peak at 414 nm which confirmed the formation of AgNPs. Fourier Transfer infrared (FTIR) confirmed the involvement of biological molecules in the formation of nanoparticles (NPs). The SEM revealed that the NPs have approximately 38 nm size. The agar well diffusion assay was used to determine antibacterial activity while tube dilution method was used to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The human pathogenic bacterial strains i.e., P. aeruginosa (MN900691) and B. subtilis (MN900684), were used as test strains. The anti-bacterial assay against test strains revealed that these NPs showed concentration dependent increased zone of inhibition (ZOI). The maximum ZOI at 25 µL of AgNPs was 20 mm against B. subtilis after 24 hours of incubation. One-way ANOVA test showed significant ZOI (p ≤ 0.05) against B. subtilis. The MIC was ranged from 4.3-6.6 μg/mL while MBC ranged from 8.3 to 6.6 μg/mL. Overall, this study suggested that the biogenically synthesized NPs are an effective alternative source of antimicrobials against pathogenic bacteria.
Bacillus licheniformis
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This study synthesized silver N-methyl chitosan (Ag-NMC) and tested it for its antimicrobial and antifungal activity. Ag-NMC was characterized by FTIR, XRD, measured for its molecular weight (MW), solubility, and toxicity. The antimicrobial activity was tested by the agar diffusion method, determining the MIC (Minimum Inhibitory Concentration), MBC (Minimum Bactericidal Concentration) against Staphylococcus aureus and Escherichia coli bacteria, and determining the Minimum Fungicidal Concentration (MFC) against the fungus Candida albicans. The results showed that Ag-NMC had MW, solubility, and LC50 of 555.65 g/mol, 50 mg/mL, 945,492 mg/L, respectively. The diameter of the inhibition zone from the resulting diffusion test showed that Ag-NMC had better antimicrobial activity than N-methyl chitosan (NMC) and chitosan. The MIC, MBC, and MFC values of Ag-NMC were always lower than that of NMC and chitosan.
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Relevance. Urinary tract infections pose a growing threat to humanity due to the rise of antibiotic resistance in uropathogens. Exploring natural sources for alternative treatments has become a prominent approach. The aim of the research was to investigate the antibacterial effects of clove (Syzygium aromaticum L.) against uropathogenic Escherichia coli (E. coli). Materials and Methods. The research was performed on three clinical multidrug-resistant uropathogenic E. coli isolates and E. coli ATCC 25922. Clove hydroalcoholic extract was obtained by cold maceration technique. To evaluate the antibacterial activity of the extract, agar well diffusion method was performed. Minimum inhibitory and minimum bactericidal concentrations of the extract were determined by microbroth dilution method. Light microscopy was used to investigate morphological changes in uropathogenic E. coli after exposure to clove extract. Checkerboard assay was used to assess synergism between clove extract and antibiotics. All obtained data were statistically processed. Results and Discussion. In well diffusion method, bacterial responses to clove extract were concentration-dependent with inhibition zone diameter of 7-10/10-15 mm for uropathogenic strains and E. coli ATCC 25922, respectively. Minimum inhibitory and minimum bactericidal concentrations of clove extract against uropathogenic strains were 25 mg/mL. The extract showed a lower minimum inhibitory concentration against E. coli ATCC 25922 (6.25 mg/ mL) with minimum bactericidal concentration being 25 mg/mL. Minimum inhibitory and minimum bactericidal concentrations ratio showed that clove extract tends to be bactericidal agent. Synergy test revealed that the combination of clove extract and nitrofurantoin or ciprofloxacin resulted in no interaction. However, minimum inhibitory concentrations of all tested agents in combinations exhibited varying degrees of decrease. Incubation of uropathogenic strains with the extract transformed them to unstable spherical L-form in percentage of 96-99 %. Conclusion. This study highlights clove as a potential natural antibacterial agent against multidrug-resistant uropathogenic E. coli, warranting further investigations into its antibacterial properties.
Syzygium
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Abstract Three different aquatic plants Selaginellaplana (Desv. Ex. Poir) Hieron, Ludwigiaadscendens (L.) H. Hara, dan Ephitemabenthamii C. B. Clarke were collected from South Sulawesi and crude extracts all of plants parts were prepared in ethanol. This extracts were checked for the presence of inhibitory zone by agar well diffusion method against gram negative Flavobacteriumcolumnare bacterial. Furthermore Minimum Inhibitory Concentration (MIC), Minimum bactericidal concentrations (MBC) testing has been done for crude extract of plants. Extracts of three plants prepared in ethanol showed relatively inhibitory zone to Flavobacterium columnareand two extracts of above plants were positive in MIC and MBC.
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Agar diffusion test
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Ralstonia solanacearum
Silver nanoparticle
Minimum bactericidal concentration
Agar diffusion test
Pathogenic bacteria
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The aim of this paper was to investigate the antibacterial potential of Kelussia odoratissima Mozff extract against Gram-negative and Gram-positive bacteria. Karafs-eKoohi with the scientific name of Kelussia odoratissima is an Iranian endemic edible plant in the middle region of Iran with enormous use as food, spice and medicinal herb . The antibacterial effect of the extracts was investigated using pour plate and disk diffusion methods. Minimum Inhibitory Concentration (MIC( and Minimum Bactericidal Concentration (MBC) were also studied using the dilution method. Repeated measure ANOVA was used for data analysis. The results showed that in disk diffusion method all concentrations of ethanolic extract had inhibitory effect against Bacillus subtilis and Staphylococcus aureus . Minimum Inhibitory Concentration (MIC( of Kelussia odoratissima leaves of aqueous and ethanolic extracts for Bacillus subtilis and Staphylococcus aureus were 16 and 8 mg/ml, and for Enterobacter aerogenes were 32 and 16 mg/ml, respectively. Minimum Bactericidal Concentration (MBC) of Kelussia odoratissima leaves of aqueous and ethanolic extracts for Bacillus subtilis and Staphylococcus aureus were 32 and 16 mg/ml, and for Enterobacter aerogenes were 64 and 32mg/ml, respectively. The results showed that the extract of Kelussia odoratissima had a satisfactory antimicrobial activity and the ethanolic extract of Kelussia odoratissima leaves had greater inhibitory effects on the strains studied compared to aqueous extract in vitro. A significant correlation was also observed between zone of inhibition and concentration of extracts.
Enterobacter aerogenes
Minimum bactericidal concentration
Agar diffusion test
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