Minimum Inhibitory Concentration Concentration Research Articles

Geraniol inhibits both planktonic cells and biofilms of the Candida parapsilosis species complex: Highlight for the improved efficacy of amphotericin B, caspofungin and fluconazole plus Geraniol.

The Candida parapsilosis species complex poses a recognized threat to the nosocomial environment. In the scenario of the global rise of resistant strains to antifungals, geraniol, a terpene isolated from different essential oils, has shown promising antimicrobial activity. We evaluated: (1) the effects of geraniol against the C. parapsilosis species complex, in planktonic and biofilm forms; (2) the strains' susceptibility to clinical antifungals and (3) the geraniol interaction with antifungals. Eighteen isolates were subjected to in vitro susceptibility testing by the broth microdilution protocol, using geraniol, amphotericin B, caspofungin, itraconazole and fluconazole to determine the minimum inhibitory concentration (MIC) and subsequently, we measured the fungicidal activity. Geraniol was tested against biofilms by the measurement of the metabolic activity and biomass. Pharmacological interactions were performed by the checkerboard method. Geraniol's MIC range was between 256 and 512µg/ml. MIC range for clinical antifungals was≤0.031-4µg/ml. Geraniol also showed antibiofilm activity with average reductions of metabolic activity (38.33%) and biomass (30.69%), at MIC concentration. Furthermore, geraniol showed synergistic/additive effects with antifungals. Briefly, geraniol inhibits both planktonic cells and biofilms of the C. parapsilosis species complex and besides it improves the efficacy of amphotericin B, caspofungin and fluconazole.

In vivo pharmacodynamic evaluation of the novel nystatin derivative BSG005 in the invasive candidiasis and invasive pulmonary aspergillosis mouse models.

Nystatin, a polyene, is one of the oldest antifungal drugs with wide in vitro potency. BSG005 is a novel, chemically modified, nystatin-like molecule in development for systemic therapy. We evaluated the pharmacokinetic/pharmacodynamic (PK/PD) relationships and target exposures using in vivo invasive pulmonary aspergillosis (IPA) and invasive candidiasis (IC) infection models for BSG005 against common fungal pathogens including Aspergillus fumigatus, Candida albicans, Candida auris, and Candida glabrata. For each species group, three to four strains were selected. Minimum inhibitory concentration (MIC) testing was done by Clinical Laboratory Standards Institute (CLSI) methods. Single-dose kinetics for BSG005 were performed at four dose levels. The immunosuppressed mouse IPA model was used for A. fumigatus studies. For all Candida studies, we utilized the neutropenic disseminated candidiasis model. We used quantitative PCR to enumerate Aspergillus in the lung and colony forming units (CFU) counts for Candida in the kidney. Treatment results were evaluated based on both area under the concentration-time curve (AUC)/MIC and maximum plasma concentration (Cmax)/MIC exposures. The BSG005 MIC was 1 mg/L against all strains. Escalating doses of BSG005 resulted in increased effect and, in general, the dose-response curves within each species were concordant. The median 96-h AUC/MIC associated with net stasis was lowest at 6.08 for C. glabrata. Increasing exposures were needed for same outcome for C. auris at 18.7, C. albicans at 29.3, and A. fumigatus at 102.4. Cmax/MIC targets for the four groups were 0.22, 0.48, 0.60, and 1.41. BSG005 demonstrated potent activity against a variety of fungal pathogens in the neutropenic mouse models. Cmax/MIC PK/PD targets were numerically lower than other polyene studies using the same infection models.

Antibacterial and antibiofilm activity of silver nanoparticles stabilized with C-phycocyanin against drug-resistant Pseudomonas aeruginosa and Staphylococcus aureus.

Biofilms are bacterial communities that can protect them against external factors, including antibiotics. In this study, silver nanoparticles (AgNPs) were formed by modifying AgNPs with C-phycocyanin (Ag-Pc) to inhibit the growth of carbapenem-resistant Pseudomonas aeruginosa (CR P. aeruginosa) and methicillin-resistant Staphylococcus aureus (MRSA) and destroy biofilm of these bacteria. The AgNPs were prepared with the green synthesis method, and Pc was used to stabilize the AgNPs. The Ag-Pc's antibacterial and antibiofilm effects were evaluated using the Microbroth dilution method and microtiter plate assay. The inhibitory effect of Ag-Pc on the expression of biofilm-related genes was evaluated by real-time PCR. Moreover, the MTT assay was used to assess the Ag-Pc toxicity. The Ag-Pc minimum inhibitory concentration (MIC) was 7.4μg/mL for CR P. aeruginosa and MRSA. Pc did not show antibacterial effects against any of the strains. Ag-Pc suppressed biofilm formation and destroyed matured biofilm in both bacteria more efficiently than the AgNPs (P< 0.05). The expression of all genes was not significantly reduced in the presence of synthesized nanoparticles. Finally, the MTT assay results did not show toxicity against a murine fibroblast cell line (L929) at MIC concentration. The present study showed the promising potential of Pc for improving the antibacterial and antibiofilm function of AgNPs and inhibiting drug-resistant bacteria. Therefore, Ag-Pc nanoparticles can be considered a promising therapeutic approach for the managing of the bacterial biofilm.

Exploring the antimicrobial efficacy of tea tree essential oil and chitosan against oral pathogens to overcome antimicrobial resistance

BackgroundConsidering that antimicrobial resistance among oral pathogens is a significant concern in dental practice, with broader implications for overall health due to the oral microbiota serving as a reservoir for antibiotic resistance genes (ARGs), research into natural products is crucial for addressing this issue. ObjectiveThis study aimed to evaluate tea tree oil (TTO) and chitosan (CH) performance against oral pathogens, including mixed-species biofilm, and its effects on bacteria growth, in addition to chemical characterization and cytotoxicity of TTO. MethodsTea Tree Oil and low molecular weight chitosan were used in this study. The chemical composition of TTO was analyzed using gas chromatography coupled with mass spectrometry (GC-MS). To evaluate TTO's antimicrobial properties, time-kill and cell viability assays were conducted. Additionally, minimum inhibitory concentration (MIC), minimum microbiocidal concentration (MMC), checkerboard, and biofilm assays were performed using TTO and CH alone and in combination. ResultsTTO chromatography peaks found consistent with the standard ISO4730:2017 and literature. TTO and CH exhibited inhibitory activity against all tested microorganisms. The predominantly microbiostatic activity of TTO is probably related to terpinen-4-ol associated with terpinene. The oil at MIC value was able to delay the log phase of Aggregatibacter actinomycetemcomitans growth. Fibroblasts (L929) viability remained above 70 % during 24 h for TTO concentrations ranging from 0.5 to 0.0625 mg/ml. TTO-CH combination showed a synergistic activity (FIC = 0.5) against A. actinomycetemcomitans and Streptococcus sanguinis, at a concentration of 0,25MIC for both species. The compounds at MIC concentration inhibited both monospecies and mixed-species biofilms studied bacteria to the same extent as the azithromycin control. ConclusionTTO and CH demonstrated efficacy in combating oral pathogens and TTO-CH combination offers a promising approach to confront microbial resistance in the oral environment.

Effects of Bacteriocin Extracted from Lactobacillus plantarum on Treatment-Resistant Escherichia coli Bacteria Isolated from Humans and Livestock

Background: Lactic acid bacteria (LAB) are among the most common probiotics and play a significant role in promoting health within the human intestinal microbiota. Their protective functions in the intestine include producing antimicrobial compounds, regulating the balance of the intestinal microbiome, and more. Objectives: This study aims to investigate the antimicrobial effects of bacteriocin extracted from Lactobacillus plantarum (L. plantarum) on Escherichia coli (E. coli) strains that cause treatment-resistant urinary tract infections (UTIs) in humans and mastitis in livestock. Methods: Twenty E. coli isolates were obtained from patients with UTIs and cows with mastitis. After culturing the samples on Eosin-methylene blue, McConkey agar, and blood agar, the strains were identified using biochemical tests. Bacteriocin produced by L. plantarum ATCC8014 was extracted, and its concentration was measured using the Bradford method. The antimicrobial effect of the bacteriocin was assessed using the broth microdilution method to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Additionally, the bacteriocin's antimicrobial activity was evaluated through three repetitions of disk diffusion and agar well diffusion methods. Results: The findings revealed that the MIC and MBC concentrations for all human and animal strains were 13.38 μg/mL and 26.76 μg/mL, respectively. Notably, two animal samples (No. 3 and No. 10) and one human sample (No. 6) showed lower MIC and MBC concentrations compared to the other samples and the standard E. coli sample. However, all strains were resistant to bacteriocin in the disk diffusion and agar well diffusion methods. Conclusions: The data indicate a high risk of increasing treatment-resistant E. coli strains in UTIs and livestock, which could pose a significant threat to public and animal health. Therefore, it is essential to further investigate the effects of different bacteriocins on these resistant strains, in conjunction with antibiotic therapies.

Combined potential of copaifera officinalis oleoresin and chitosan against oral pathogens.

Copaifera officinalis Oleoresin (COR) and Chitosan (CH) were combined to test the potential to inhibit oral bacteria. First, COR was analyzed by GC-MS to identify its main constituents and then Minimum Inhibitory Concentration (MIC) assays and Minimum Microbiocidal Concentration (MMC) of the compounds alone against 17 pathogens were performed. Sixteen primary compounds were identified in COR, but the major constituent was β-Caryophyllene (40.5%). COR showed MIC concentrations of 26.04 to 46.87µg/mL and CH 0.1mg/mL to 0.8mg/mL. Second, the combination against oral bacteria strains was tested using a checkerboard test with the determination of Fractional Inhibitory Concentration (FIC) for synergistic effect, followed by the bacterial biofilm aggregation test using monospecies and mixed biofilm. The combination of COR and CH showed a synergistic effect for S. oralis (ATCC 10557) and an additive effect for the other strains tested, promoting bactericidal activity, as well as reducing the concentrations needed to cause bacterial inhibition. In addition, it showed good activity in inhibiting biofilm formation, with inhibition percentages close to Azithromycin. The results of this study highlight the synergistic potential of COR and CH combination as a promising strategy in the search for innovative antimicrobial therapies for infections related to oral bacterial biofilms.

Comparative Evaluation of Antimicrobial Efficacy of Silver Nanoparticles Infused with Azadirachta indica Extract and Chlorhexidine Against Red-complex Pathogens.

The present study aimed to evaluate the antimicrobial efficacy of silver nanoparticles infused with Azadirachta indica extract and chlorhexidine against red-complex periopathogens. Neem leaf extraction was done followed by standardization to the synthesis of neem-infused silver nanoparticles and fractionation of compounds done by using thin layer chromatography to separate the mixture of neem leaf extract. Characterization of neem-infused silver nanoparticles was done by scanning electron microscopy and UV-Visible spectroscopy. The compound identified in neem-infused silver nanoparticles was gedunin which was confirmed by Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. Determination of antibacterial activity done by disc diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) methods. Group I-99% ethanolic extract, group II-neem-infused silver nanoparticles (NAgNPs), group III-chlorhexidine. The relative inhibitory zone value for Tannerella forsythia (180) in neem-infused silver nanoparticles (group II) was greater when compared with other periopathogens Porphyromonas gingivalis (133) and Treponema denticola (160) than 99% ethanolic extract (group I), chlorhexidine (group III). Neem-infused silver nanoparticles (group III) showed superior antimicrobial activity against T. forsythia (19.3 ± 31.1547) and T. denticola (18±0) when compared with P. gingivalis (17.6 ± 0.5774). On evaluating MIC and minimum bacterial concentrations, P. gingivalis is more resistant than other pathogens in neem-infused silver nanoparticles (group III). Neem-infused silver nanoparticles exhibited superior antibacterial activity as compared with gold-standard chlorhexidine against red-complex periodontal pathogens. For MIC and MBC all the three periopathogens were effective but P. gingivalis was more resistant. Antibiotics are effective against many drug-resistant bacteria. As a ready-made medicine, they can be used to treat many infections. Silver nanoparticles in drug delivery systems generally increase solubility, stability, and biodistribution, thereby increasing their effectiveness. Green synthesis using plant extracts as precursors to synthesize nanoparticles has proven to be environmentally non-hazardous combined with remarkably improved efficacy against bacterial and viral diseases. So neem-infused silver nanoparticles can be utilized as a drug delivery system. Hence, it can be used as a potential antibacterial ingredient in formulations for periodontal use like mouthwashes and gels for local drug delivery. How to cite this article: Krishnappan S, Ravindran S, Balu P, et al. Comparative Evaluation of Antimicrobial Efficacy of Silver Nanoparticles Infused with Azadirachta indica extract and Chlorhexidine Against Red-Complex Pathogens. J Contemp Dent Pract 2024;25(6):547-553.

Antibacterial and antibiofilm activities of tribulus terrestris methanolic extract against Streptococcus mutans, Streptococcus sobrinus, and Lactobacillus acidophilus: An in vitro study

ABSTRACT Background: Tribulus terrestris (TT) extract has shown good antibacterial activity against some bacteria. However, there are limited data on its cariogenic properties. This in vitro study aimed to evaluate the antibacterial and antibiofilm activities of TT extract against Streptococcus mutans (S. mutans), Streptococcus sobrinus (S. sorbinus), and Lactobacillus acidophilus (L. acidophilus) as the important cariogenic bacteria. Materials and Methods: This study was designed in an experimental model (in vitro). Phytochemical tests were carried out to detect herbal compounds in the TT extract. Agar well diffusion was performed to compare the extract (500–62.5 mg/mL) with different concentrations of chlorhexidine (2–0.25 mg/mL). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the TT extract and chlorhexidine were also determined. The lowest concentration showing ≥50% inhibition of biofilm formation (MBIC50) was determined using crystal violet assay. Further, the time-kill assay (Log of CFU/mL) was performed, and acid production (pH) was measured at 1 × MIC concentration in 2, 4, 8, 12, and 24 h. Data analysis conducted using SPSS software (v26, IBM) involved One-way analysis of variance, Tukey post hoc tests, and t-test to compare concentrations and groups. Significance level is set at 0.05. Results: The TT extract mostly consisted of flavonoids. Its inhibition zones in the well diffusion test were statistically comparable with chlorhexidine in some concentrations (P &gt; 0.05). The MIC of the TT extract was 15.625 mg/mL for all tested bacteria, whereas the MBC ranged from 31.25 to 62.5 mg/mL. Further, the MBIC50 ranged from 7.8125 to 15.625 mg/mL for the extract. Time-kill assay showed that the bactericidal activity of the TT extract lasted for 8, 12, and 2 h for S. mutans, S. sobrinus, and L. acidophilus, respectively. The acid production decreased obviously after 8 h. Conclusion: The TT extract showed good time-dependent antibacterial and antibiofilm activity, as well as acid production inhibition, against cariogenic bacteria in laboratory experiments.

Antifungal activity of Cinnamaldehyde derivatives against fluconazole-resistant Candida albicans

BackgroundCandida albicans is an opportunistic pathogen commonly found in human mucous membranes. In light of the escalating challenge posed by antibiotic resistance of C. albicans strains worldwide, it is an urgently necessary to explore alternative therapeutic options. ObjectiveThis study aims to assess the efficacy of two Cinnamaldehyde derivatives, 2-Cl Cinnamaldehyde (2-Cl CA) and 4-Cl Cinnamaldehyde (4-Cl CA), against C. albicans through both in vitro experiments and in vivo murine models and to evaluate their potential as new drug candidates for treating C. albicans. Methods and resultsThe minimum inhibitory concentrations (MICs) of Cinnamaldehyde 2-Cl and 4-Cl benzene ring derivatives against C. albicans were 25 μg/mL. Time-killing experiments revealed that both Cinnamaldehyde derivatives exhibited fungicidal activity against C. albicans at concentrations of 5 MIC and 10 MIC. In the checkerboard experiment, 4-Cl CA did not show any antagonistic effect when combined with first-line antifungal drugs. Instead, it exhibited additive effects in combination with nystatin. Both 2-Cl and 4-Cl CA demonstrated inhibitory activity against C. albicans biofilm formation, especially at 8 MIC and 16 MIC concentrations. In C. albicans biofilm eradication experiments, although high drug concentrations of 2-Cl and 4-Cl CA were unable to eradicate the biofilm completely, they were still effective in killing C. albicans cells within the biofilm. Moreover, sub-inhibitory concentrations of 4-Cl CA (ranging from 5 to 20 μg/mL) significantly inhibited cell aggregation and hyphal formation. Furthermore, 4-Cl CA effectively inhibited intracellular C. albicans infection in macrophages. Lastly, the effectiveness of 4-Cl CA was evaluated in a mouse model of hematogenous disseminated candidiasis caused by C. albicans, which revealed that 4-Cl CA significantly reduced fungal burden and improved mouse survival compared to the untreated controls. ConclusionThe 4-Cl CA exhibited inhibitory effects against C. albicans through both in vivo and in vitro models, demonstrating its therapeutic potential as a promising new drug candidate for treating drug-resistant candidiasis albicans.

Antimicrobial resistance induction potential of grapefruit seed extract on multi-species biofilm of E. coli in food industry

Biofilm formation in natural environments involving complex multi-structural arrangements hinders challenges in antimicrobial resistance. This study investigated the antimicrobial resistance potential of grapefruit seed extract (GSE) by examining the formation of mono-, dual-, and multi-species biofilms. We also explored the counterintuitive effect in response to GSE at various concentrations, including minimum inhibitory concentration (MIC) and sub-MIC (1/2 and 1/4 MIC). The results of the swimming and swarming motility tests revealed increased motility at the sub-MIC of GSE. The crystal violet assay demonstrated increased biofilm formation in multi-species biofilms, highlighting the synergistic effect of Escherichia coli, Salmonella Typhimurium, and Listeria monocytogenes. At the MIC concentration of GSE, field emission scanning electron microscopy (FE-SEM) revealed cell morphology damage, while sub-MIC increased biofilm formation and architectural complexity. Multi-species biofilms demonstrated greater biofilm-forming ability and antimicrobial resistance than mono-species biofilms, indicating synergistic interactions and enhanced resilience. These findings highlight the importance of understanding biofilm dynamics and antimicrobial resistance to ensure environmental safety.

Ratanjot (Alkanna tinctoria L.) Root Extract, Rich in Antioxidants, Exhibits Strong Antimicrobial Activity against Foodborne Pathogens and Is a Potential Food Preservative.

Natural and sustainable plant-based antioxidants and antimicrobials are highly desirable for improving food quality and safety. The present investigation assessed the antimicrobial and antioxidant properties of active components from Alkanna tinctoria L. (herb) roots, also known as Ratanjot root. Two methods were used to extract active components: microwave-assisted hot water (MAHW) and ethanolic extraction. MAHW extract yielded 6.29%, while the ethanol extract yielded 18.27%, suggesting superior Ratanjot root extract powder (RRP) solubility in ethanol over water. The ethanol extract showed significantly higher antioxidant activity than the MAHW extract. Gas Chromatography-Mass Spectrometry analysis revealed three major phenolic compounds: butanoic acid, 3-hydroxy-3-methyl-; arnebin 7, and diisooctyl pthalate. The color attributes (L*, a*, b*, H°ab, C*ab) for the ethanolic and MAHW extracts revealed significant differences (p < 0.05) in all the above parameters for both types of extracts, except for yellowness (b*) and chroma (C*ab) values. The ethanol extract exhibited antimicrobial activity against 14 foodborne bacteria, with a significantly higher inhibitory effect against Gram-positive bacteria (Listeria monocytogenes and Staphylococcus aureus) than the Gram-negative bacteria (Salmonella enterica serovar Typhimurium and Escherichia coli). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were both 25 mg/mL for the Gram-negative bacteria, while the MIC and MBC concentrations varied for Gram-positive bacteria (0.049-0.098 mg/mL and 0.098-0.195 mg/mL) and the antimicrobial effect was bactericidal. The antimicrobial activities of RRP extract remained stable under broad temperature (37-100 °C) and pH (2-6) conditions, as well as during refrigerated storage for 30 days. Application of RRP at 1% (10 mg/g) and 2.5% (25 mg/g) levels in a cooked chicken meatball model system prevented lipid oxidation and improved sensory attributes and retarded microbial growth during refrigerated (4 °C) storage for 20 days. Furthermore, the RRP extract was non-toxic when tested with sheep erythrocytes and did not inhibit the growth of probiotics, Lacticaseibacillus casei, and Lactiplantibacillus plantarum. In conclusion, the study suggests that RRP possesses excellent antimicrobial and antioxidant activities, thus making it suitable for food preservation.

Lavandula angustifolia Essential Oil Inhibits the Ability of Fusobacterium nucleatum to Produce Volatile Sulfide Compounds, a Key Components in Oral Malodor.

Oral malodor still constitutes a major challenge worldwide. A strong effort is invested in eliminating volatile sulfur compound-producing oral bacteria through organic natural products such as essential oils. Fusobacterium nucleatum is a known volatile sulfur compound-producing bacteria that inspires oral malodor. The aim of the present study was to test the effect of lavender essential oil on the bacterium's ability to produce volatile sulfide compounds, the principal components of oral malodor. Lavender (Lavandula angustifolia) essential oil was extracted by hydrodistillation and analyzed using GC-MS. The minimal inhibitory concentration (MIC) of lavender essential oil on Fusobacterium nucleatum was determined in a previous trial. Fusobacterium nucleatum was incubated anaerobically in the presence of sub-MIC, MIC, and above MIC concentrations of lavender essential oil, as well as saline and chlorhexidine as negative and positive controls, respectively. Following incubation, volatile sulfur compound levels were measured using GC (Oralchroma), and bacterial cell membrane damage was studied using fluorescence microscopy. Chemical analysis of lavender essential oil yielded five main components, with camphor being the most abundant, accounting for nearly one-third of the total lavender essential oil volume. The MIC (4 µL/mL) of lavender essential oil reduced volatile sulfur compound secretion at a statistically significant level compared to the control (saline). Furthermore, the level of volatile sulfur compound production attributed to 1 MIC of lavender essential oil was in the range of the positive control chlorhexidine with no significant difference. When examining bacterial membrane damage, 2 MIC of lavender essential oil (i.e., 8 µL/mL) demonstrated the same, showing antibacterial membrane damage values comparative to chlorhexidine. Since lavender essential oil was found to be highly effective in hindering volatile sulfur compound production by Fusobacterium nucleatum through the induction of bacterial cell membrane damage, the results suggest that lavender essential oil may be a suitable alternative to conventional chemical-based anti-malodor agents.

Antimicrobial and Membrane Stabilization Activity of Plant Extracts Against Pathogenic Bacteria and Fungi

The use and search for antimicrobial drugs derived from plants have accelerated in recent years. The present study was aimed to determine the antimicrobial, antibiofilm and membrane stabilization activities of plant extracts. Crude extracts of four plants namely, Mikania scandens (L.), Mimosa pudica (L.), Murraya paniculata (L.), and Syzygium aromaticum (L.) were tested against eleven human pathogens including four biofilm producing bacterial strains Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus, and one fungal strain Candida albicans. The antimicrobial activities as well as minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extracts were evaluated using disc diffusion and macro-broth dilution methods respectively. Among the plant extracts, ethanol extract of S. aromaticum exhibited the largest zone of inhibition 35 mm in diameter against Bacillus subtilis at 500 µg/disc concentrations. The lowest MIC (1000 µg/ml) and MBC (2500 µg/ml) were determined against P. aeruginosa with the same extract of S. aromaticum. In case of time kill assay, S. aromaticum extract showed the lowest optical density (OD600) 0.2 against E coli in 3h at MBC concentration. Moreover, the same extract of S. aromaticum displayed the strong antibiofilm activity, inhibiting 100% biofilm formation of E. coli at 2×MIC concentration. Furthermore, in vitro experiment was performed to evaluate membrane stabilization activity of plant extracts and S. aromaticum showed 100% activity in stabilizing cell membranes, preventing hemolysis of red blood cells. Therefore, this study provides valuable insight for designing antimicrobial products to efficiently eliminating human infections, and the plant extracts could be the potential antimicrobial agent. Bangladesh J Microbiol, Volume 40, Number 2, December 2023, pp 76-85

Mechanism of antibacterial activity of diallyl sulfide against Bacillus cereus

World health organization (WHO) recognizes antimicrobial resistance as a silent pandemic. It is estimated that 10 million deaths will occur annually due to antimicrobial resistant infections by 2050. Phytochemicals exhibit activity against drug resistant bacteria, offering potential for developing novel antibacterial agents. Garlic organosulphur compounds exhibit potent activity against a variety of drug-resistant bacteria. Identifying their mechanism of action is critical to assess their potential to be developed as novel antibacterial agents. Diallyl sulfide (DAS) is a component of garlic essential oil with antibacterial activity. In this study antibacterial activity of DAS was investigated against Bacillus cereus, a common foodborne pathogen. DAS exhibited activity against B. cereus with a minimum inhibitory concentration (MIC) of 54.75 mM. The presence of DAS significantly reduced the growth of B. cereus. The study also investigated the mechanism of antibacterial activity of DAS against B. cereus. Treating B. cereus with sub-MIC and MIC concentration of DAS resulted in a dose and time-dependent leakage of intracellular proteins. The protein leakage was enhanced at acidic pH. Scanning electron microscopy (SEM) of B. cereus treated with DAS showed deformation in the cell membrane. Thus, the data indicate that DAS exerts its antibacterial activity by compromising the membrane integrity of B. cereus. The study demonstrates DAS could be used to control B. cereus infections. The findings indicate that DAS has a membrane altering activity, suggesting that development of resistance to this mechanism is less likely and the compound could be novel antibacterial or a good adjuvant for antibiotics.

Analysis of the Molecular Structure of Hydroxychavicol, a Promising Oral Antibacterial

Abstract Objectives In order to better understand hydroxychavicol’s effectiveness as an oral antibacterial, its structural components were analyzed with respect to minimum inhibitory concentrations and minimum bactericidal concentrations against various oral bacteria. These structural components include the free hydroxy groups and allyl chain connected to hydroxychavicol’s benzene core. Methods Six structural analogs of hydroxychavicol were tested against a range of oral bacteria using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. MIC results were obtained using serial microdilution techniques in 96-well plates with resazurin dye as a colorimetric indicator. Aliquots within each MIC concentration range were then placed on appropriate agar medium and the minimum bactericidal concentration was determined as the lowest concentration with no observed colony growth. Key Findings A synergistic interaction was observed between the allyl chain and hydroxy groups on the benzene core of hydroxychavicol, which resulted in lower MICs against the tested oral bacteria. It was also found that a hydroxy group para to the allyl chain on the benzene ring resulted in more effective inhibition, with a MIC of &amp;lt;50 μg/mL against R. dentocariosa. Additionally, analytes possessing free hydroxy groups ortho to one another on the benzene ring resulted in MICs of 200-300 μg/mL or lower, whereas analytes with free hydroxy groups meta to one another on the benzene ring exhibited MICs of &amp;gt;1000 μg/mL. Conclusions This study helps elucidate the structural components responsible for hydroxychavicol’s effectiveness as an oral antibacterial. The findings herein help to understand the mechanism of hydroxychavicol’s antibacterial properties and will be helpful in the design and synthesis of more effective oral antibacterial treatments.

Interaction between Enrofloxacin and Three Essential Oils (Cinnamon Bark, Clove Bud and Lavender Flower)-A Study on Multidrug-Resistant Escherichia coli Strains Isolated from 1-Day-Old Broiler Chickens.

Avian pathogenic Escherichia coli (APEC) causes a variety of infections outside the intestine. The treatment of these infections is becoming increasingly difficult due to the emergence of multi-drug resistant (MDR) strains, which can also be a direct or indirect threat to humans as consumers of poultry products. Therefore, alternative antimicrobial agents are being sought, which could be essential oils, either administered individually or in interaction with antibiotics. Sixteen field isolates of E. coli (originating from 1-day-old broilers) and the ATCC 25922 reference strain were tested. Commercial cinnamon bark, clove bud, lavender flower essential oils (EOs) and enrofloxacin were selected to assess the sensitivity of the selected E. coli strains to antimicrobial agents. The checkerboard method was used to estimate the individual minimum inhibitory concentration (MIC) for each antimicrobial agent as well as to determine the interactions between the selected essential oil and enrofloxacin. In the case of enrofloxacin, ten isolates were resistant at MIC ≥ 2 μg/mL, three were classified as intermediate (0.5-1 μg/mL) and three as sensitive at ≤0.25 μg/mL. Regardless of the sensitivity to enrofloxacin, the MIC for cinnamon EO was 0.25% v/v and for clove EO was 0.125% v/v. All MDR strains had MIC values for lavender EO of 1% v/v, while drug-sensitive isolates had MIC of 0.5% v/v. Synergism between enrofloxacin and EO was noted more frequently in lavender EO (82.35%), followed by cinnamon EO (64.7%), than in clove EO (47.1%). The remaining cases exhibited additive effects. Owing to synergy, the isolates became susceptible to enrofloxacin at an MIC of ≤8 µg/mL. A time-kill study supports these observations. Cinnamon and clove EOs required for up to 1 h and lavender EO for up to 4 h to completely kill a multidrug-resistant strain as well as the ATCC 25922 reference strain of E. coli. Through synergistic or additive effects, blends with a lower than MIC concentration of enrofloxacin mixed with a lower EO content required 6 ± 2 h to achieve a similar effect.

Investigation of the Antibacterial Synergistic Activity of Fosfomycin-Teicoplanin Combination in Methicillin-Resistant Staphylococcus aureus Strains Isolated from Various Clinical Sample

The aim of this study was to investigate the detection of teicoplanin and fosfomycin antibiotic susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) strains by different methods and to evaluate the antibacterial synergistic effect of teicoplanin-fosfomycin combination by using checkerboard assay and time kill curve assay. Forty-five MRSA strains isolated from clinical samples in routine medical microbiology laboratory of Göztepe Prof. Dr. Süleyman Yalçın City Hospital were included in the study. In the first stage of the combination study, minimum inhibitory concentrations (MIC) were investigated by broth microdilution for teicoplanin and by both broth microdilution and agar dilution methods for fosfomycin. The combination of teicoplanin and fosfomycin was tested by the checkerboard method in 45 MRSA strains and combination effect was determined according to fractional inhibitory concentration index (ΣFIC) calculation. The synergistic effect and bactericidal activity of antibiotic combination were studied against a randomly selected strain from the strains used in the study by using time-kill method for 24 hours. As a result of teicoplanin and fosfomycin antibiotic susceptibility studies, all isolates were found to be susceptible to both antibiotics according to the susceptibility breakpoints determined by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). A synergistic effect was found in 22 (49%), additive effect in 22 (49%) and indifferent effect in one (2%) of the 45 strains studied with the checkerboard method. The mean ΣFIC of 45 isolates was found to be 0.5. In the combination study of the antibiotics of the isolate that was studied with time-kill method, synergism was detected for 1/8 MIC concentrations at 12th hour and 24th hour and synergism at 1/4 MIC concentration at sixth hour, 12th hour and 24th hour. In the combination study of 1/4 MIC concentrations of antibiotics, bactericidal effect was detected at sixth hour and this effect was observed to disappear at 12th and 24th hours. High rate of synergistic antibacterial effect of teicoplanin-fosfomycin combination on MRSA isolates was demonstrated as a result of in vitro tests. Such studies conducted on antibiotic-resistant bacterial infections will provide clinicians different treatment options and will contribute to increasing survival. As a result of this study, provided that it is supported by future clinical studies, it can be stated that the teicoplanin-fosfomycin combination may be an effective treatment option in community and hospital-acquired infections caused by MRSA.

Equinins as Novel Broad-Spectrum Antimicrobial Peptides Isolated from the Cnidarian Actinia equina (Linnaeus, 1758).

Sea anemones are valuable for therapeutic research as a diversified source of bioactive molecules, due to their diverse bioactive molecules linked to predation and defence mechanisms involving toxins and antimicrobial peptides. Acid extracts from Actinia equina tentacles and body were examined for antibacterial activity against Gram-positive, Gram-negative bacteria, and fungi. The peptide fractions showed interesting minimum inhibitory concentration (MIC) values (up to 0.125 µg/mL) against the tested pathogens. Further investigation and characterization of tentacle acid extracts with significant antimicrobial activity led to the purification of peptides through reverse phase chromatography on solid phase and HPLC. Broad-spectrum antimicrobial peptide activity was found in 40% acetonitrile fractions. The resulting peptides had a molecular mass of 2612.91 and 3934.827 Da and MIC ranging from 0.06 to 0.20 mg/mL. Sequencing revealed similarities to AMPs found in amphibians, fish, and Cnidaria, with anti-Gram+, Gram-, antifungal, candidacidal, anti-methicillin-resistant Staphylococcus aureus, carbapenemase-producing, vancomycin-resistant bacteria, and multi-drug resistant activity. Peptides 6.2 and 7.3, named Equinin A and B, respectively, were synthesized and evaluated in vitro towards the above-mentioned bacterial pathogens. Equinin B exerted interesting antibacterial activity (MIC and bactericidal concentrations of 1 mg/mL and 0.25 mg/mL, respectively) and gene organization supporting its potential in applied research.

Antimicrobial photodynamic effect of the photosensitizer riboflavin, alone and in combination with colistin, against pandrug-resistant Pseudomonas aeruginosa clinical isolates

IntroductionDevelopment of multi-, extensively-, and pandrug-resistant (MDR, XDR, and PDR) strains of Pseudomonas aeruginosa remains a major problem in medical care. The present study evaluated the effect of antimicrobial photodynamic therapy (aPDT) as a monotherapy and in combination with colistin against P. aeruginosa isolates. MethodsTwo P. aeruginosa isolates recovered from patients with respiratory tract infections were examined in this study. Minimum inhibitory concentration (MIC) of colistin was determined by the colistin broth disk elution (CBDE) and the reference broth microdilution (rBMD) methods. aPDT was performed using the photosensitizer (Ps) riboflavin at several concentrations and a light-emitting diode (LED) emitting blue light for different irradiation times with or without colistin at 1/2 × MIC concentration. ResultsBoth PA1 and PA2 isolates were identified as colistin-resistant P. aeruginosa with a MIC ≥4 μg/mL by the CBDE and MICs of 512 μg/mL and 256 μg/mL, respectively, by the rBMD. In aPDT, neither riboflavin nor LED light alone had antibacterial effects. The values of colony forming units per milliliter (CFU/mL) in both isolates were significantly reduced by LED + Ps treatments in a time-dependent manner (LED irradiation time) and dose-dependent manner (Ps concentration). In comparison with control, treatment with Ps (50 μM) + LED (120 s) and Ps (100 μM) + LED (120 s) resulted in 0.27 log10 CFU/mL and 0.43 log10 CFU/mL reductions in PA1, and 0.28 log10 CFU/mL and 0.34 log10 CFU/mL reductions in PA2, respectively, (P < 0.01). The best results were obtained after the combination of aPDT followed by colistin, which increased bacterial reduction, resulting in a 0.41−0.7 log10 CFU/mL reduction for PA1 and 0.35−0.83 log10 CFU/mL reduction for PA2 (P = 0.001). ConclusionsThis study suggests the potential implications of aPDT in combination with antibiotics, such as colistin for treatment of difficult-to-treat P. aeruginosa infections.

Nanoformulated meloxicam and rifampin: inhibiting quorum sensing and biofilm formation in Pseudomonas aeruginosa.

Background: We aimed to investigate the simultaneous effects of meloxicam and rifampin nanoformulations with solid lipid nanoparticle (SLN) and nanostructured lipid carrier (NLC) substrates on inhibiting the quorum-sensing system of Pseudomonas aeruginosa and preventing biofilm formation by this bacterium. Methods: Antimicrobial activity of rifampin and meloxicam encapsulated with SLNs and NLCs against P. aeruginosa PAO1 was assessed by disk diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Results: The SLN formulation was associated with lower doses for the MIC and minimum bactericidal concentration in comparison to NLC. Moreover, our results demonstrated that both nanoformulations were able to produce 100% inhibition of the biofilm formation of P. aeruginosa PAO1. Conclusion: All these findings suggest that meloxicam and rifampin encapsulated with SLNs could be the most effective formulation against P. aeruginosa.