Antibacterial Efficacy of Mineral Trioxide Aggregate, Bioactive Glass and Epoxy-Resin Based Sealer: An In-Vitro Interventional Study.

Aim: The aim and objective of this in vitro study was to evaluate the antibacterial efficacy of mineral trioxide aggregate, bioactive glass sealer, and epoxy-resin-based sealer. Materials and Methods: In the present study, 22 Mueller Hinton agar (MH agar) plates were employed and equally divided into two groups. Three holes were made by removal of agar at equidistant points and filled with root canal sealers. The strains of the bacteria used in this study were S. aureus (ATCC 6538) and C. albicans (ATCC 10231) and were divided into two groups and root canal sealers were divided into three subgroups: mineral trioxide aggregate (MTA) fillapex Sealer, Nishika Bioactive Glass sealer, and Syntex Epoxy Resin base sealer. For Staphylococcus aureus, peptone water was placed in a 2 mL test tube and bacteria were extracted from blood agar plates using a nichrome wire loop and poured into the peptone water-containing test tube and incubated for 2 hours and for C. albicans, fungi were grown at 37°C for 24 hours in MH Broth and seeded into MH agar to produce turbidity of 0.5 on the McFarland scale, which corresponds to a concentration of 108 CFU/mL. This MH broth was used as a second layer. The seeded agar was then added over the plates immediately after the insertion of sealer cement. After incubation, the diameters of zones of inhibition around the plates were measured. Results: The results of this study showed that the highest inhibition was recorded in Syntex sealer against Staphylococcus aureus followed by MTA fillapex sealer and Nishika sealer, whereas MTA fillapex showed the highest inhibition against C. albicans followed by Syntex sealer and Nishika sealer. Conclusion: Syntex sealer exhibits better antibacterial efficacy against Staphylococcus aureus and MTA fillapex exhibit better antibacterial efficacy against C. albicans.

Accurate estimation of the inhibition zone of antibiotics based on laser speckle imaging and multiple random speckle illumination.

The antimicrobial susceptibility test (AST) plays a crucial role in selecting appropriate antibiotics for the treatment of bacterial infections in patients. The diffusion disk method is widely adopted AST method due to its simplicity, cost-effectiveness, and flexibility. It assesses antibiotic efficacy by measuring the size of the inhibition zone where bacterial growth is suppressed. Quantification of the zone diameter is typically achieved using tools such as rulers, calipers, or automated zone readers, as the inhibition zone is visually discernible. However, challenges arise due to inaccuracies stemming from human errors or image processing of intensity-based images. Here, we proposed a bacterial activity-based AST using laser speckle imaging (LSI) with multiple speckle illumination. LSI measures a speckle pattern produced by interferences of scattered light from the sample; therefore, LSI enables the detection of variation or movement within the sample such as bacterial activity. We found that LSI with multiple speckle illuminations provides consistent and uniform analysis of measured time-varying speckle images. Furthermore, our proposed method effectively identified the boundary of the inhibition zone using the k-means clustering algorithm, exploiting a result of speckle pattern analysis as features. Collectively, the proposed method offers a versatile analytical tool in the diffusion disk method.

Assessing the Efficiency of Antimicrobial Plant Extracts from Artemisia afra and Eucalyptus globulus as Coatings for Textiles.

This study aimed to assess the antimicrobial activities of plant extracts from Artemisia afra and Eucalyptus globulus when used as coatings for textiles. A pulsed ultrasound-assisted extraction method (PUAE) was employed to obtain methanolic and hexanoic extracts from both plants. Eucalyptus globulus methanol extraction exhibited the highest yield at 22.76% (±0.61%), while Artemisia afra demonstrated lower yields. Phytochemical screening identified various secondary metabolites in the extracts, including phenols, quinones, and steroids. Antimicrobial tests against Staphylococcus aureus and Escherichia coli revealed varying degrees of susceptibility, with Eucalyptus globulus hexanoic extracts showing the highest activity against Staphylococcus aureus at an average percentage growth of 18.74% (±0.26%). Minimum inhibitory concentration (MIC) values were determined for the extracts, but complete inhibition did not occur at concentrations below 500 µg/mL. The extracts exhibited varying effects on Staphylococcus aureus and Escherichia coli growth, with some extracts promoting bacterial growth. Coating textiles with Eucalyptus globulus methanolic extracts demonstrated antibacterial activity against Staphylococcus aureus with the highest zone of inhibition observed in cotton-coated samples (258.4 mm2). Polyester-coated samples exhibited smaller inhibition zones, with the lowest observed in Eucalyptus globulus methanolic extract coating (65.97 mm2). Scanning electron microscope (SEM) analysis revealed visible surface morphology changes in coated fabrics, depicting fine, cluster, lumpy, flaky, and fragment-like morphologies. Laundering effects on coated fabrics were investigated, showing a significant decrease in antimicrobial activity after washing. Fourier-transform infrared spectroscopy (FTIR) identified functional groups in the extracts associated with antimicrobial properties. The complexity of the bioactive compounds suggests potential antimicrobial efficacy, resting on factors such as geographical location, climate, and extraction methods. Notwithstanding the limitations, this study contributes valuable insights into the use of plant extracts as antimicrobial coatings for textiles.

Biosynthesis of Zinc Nanoparticles From Actinobacterium Streptomyces Species and Their Biological Potential.

BACKGROUND: In today's world, antibiotic-resistant microorganisms are a major concern. There is solid evidence that metal nanoparticles (NPs) tend to have antimicrobial properties. The most effective substitute for antibiotic resistance is the incorporation of metal NPs. The antibacterial properties of NPs are currently being explored and shown to be successful. Zinc (Zn) NPs that are biosynthesized from marine Actinobacterium proved to be more biocompatible, bioactive, and affordable.  Aim: This study aims to investigate the synthesis of ZnNPs from Actinobacterium Streptomyces species and their antimicrobial effects against gram-positive and gram-negative bacteria. MATERIALS AND METHODS: The current study uses natural, considerably safer processes to synthesize ZnNPs from marine Actinobacteria with little to no negative side effects. It involves sample collection, identification, and isolation of Actinobacterium Streptomyces species. The isolated sample was air-dried, and extracts of ZnNPs were taken. Among the isolates from marine sediment, two Actinobacteria that generate bioactive secondary metabolites-Streptomyces species (MOSEL-ME28) and Rhodococcus rhodochrous (MOSEL-ME29)-were selected for extracellular synthesis of ZnNPs. The antimicrobial activity of the biosynthesized ZnNPs from marine Actinobacteria was analyzed against Staphylococcus (MRSA), Klebsiella pneumoniae, and Streptococcus mutans. The results were statistically analyzed and graphs were created. RESULTS: ZnNPs obtained from Actinobacterium Streptomyces species exhibited antimicrobial effects against Staphylococcus (MRSA), Klebsiella, and Streptococcus mutans. At 280 nm wavelength, analysis of the UV spectrum showed a notable absorbance value of 1.8. The antibacterial efficacy against Staphylococcus MRSA, Klebsiella species, and Streptococcus mutans was assessed by measuring the zone of inhibition in diameter. The zones of inhibition were 8, 8, and 7 mm on the evaluation for Streptococcus mutans, S. aureus, and Klebsiella species, respectively, at a dose of 75 µg/mL. When the dosage was increased to 100 µg/mL, the inhibition zones were found to be 9.5, 9, and 7.5 mm for the respective bacterial strains. CONCLUSION: ZnNPs are biosynthesized from marine Actinobacterium Streptomyces species in this research study. They have a significant antimicrobial activity against both gram-positive and negative bacteria. This indicates that ZnNPs have enormous antimicrobial potential and have an extensive spectrum of applications. However, clinical trials must be completed before it can be used safely on patients.

Gallic Acid Derived 1, 2-Diarylindole as a Potential Antifungal Agent against Candida Strain.

BACKGROUND: In recent decades, Candida albicans has become a serious issue for public health. The worldwide rapid rise in drug resistance to conventional therapies is the main contributing reason. Moreover, because of their potent activity at low concentrations and apparent lack of toxicity, compounds originating from plants are used in therapeutic treatments because of their potent activity at low concentrations and apparent lack of toxicity. Particularly in immunocompromised people, Candida species can result in a wide range of ailments. OBJECTIVES: Present manuscript describes antifungal activity of an indole derivative 1-(4-((5- methoxy-2-(3,4,5-trimethoxyphenyl)-1H-indol-1-yl) methyl) phenoxy)-N,N-dimethylethan-1- amine (7, 100DL-6) by using an in-silico and in-vitro anti-candidal activity against two Candida strains; Candida kefyr-DS-02 (ATCC-204093) and Candida albicans (AI-clinical isolate, AIIMS- Delhi). METHODS: The synthetic strategy for the preparation of indole derivatives was modified through Fischer indole reaction. Antifungal activity of an indole derivative 1-(4-((5-methoxy-2-(3,4,5- trimethoxyphenyl)-1H-indol-1-yl) methyl) phenoxy)-N,N-dimethylethan-1-amine (7, 100DL-6) was done by using an in-silico and in-vitro anti-candidal activity against two Candida strains; Candida kefyr-DS-02 (ATCC-204093) and Candida albicans (AI-clinical isolate, AIIMS-Delhi). Compound 100DL-6 efficacy was determined by Combination synergy study, ergosterol binding assay, MTT toxicity study and Mutagenicity. RESULTS: Compound 100DL-6 was obtained in 65% yield on desired motifs. Docking scores found were 100DL-6 (-8.7 kcal/mol) and Fluconazole (-7.6 kcal/mol). Further, RMSD were shown for 100DL6 (0.26 ± 0.23 nm) and fluconazole (1.2 ± 0.62 nm). Indole derivative 100DL-6 was active against the tested fungal pathogens and the total zone of inhibition was measured between 13-14 mm in diameter and MIC values between 31.25 µg/mL to 250 µg/mL and MFC values between 62.5 µg/mL to 500 µg/mL. In checkerboard assay synergistic mode of interaction of 100DL-6 with known antifungal drugs was observed. In the presence of ergosterol 100DL-6 and standard drug (s) increased their MIC values, demonstrating a considerable affinity for ergosterol. Compound 100DL-6 was considered to be less-cytotoxic to the cells as determined by MTT assay. Lead compound 100DL-6 was found to be non-mutagenic. CONCLUSION: In the present study, 100DL6 (indole derivatives) significantly abrupted the ergosterol biosynthetic pathway and showed moderate anti-candidal effects. These studies suggest that 100DL6 significantly enhances antifungal effect of clinical drug fluconazole synergistically and may be considered as in clinical trial prior to some extensive in-vivo validations.

Azadirachta indica (AI)leaf extract coated ZnO-AInanocore-shell particles for enhanced antibacterial activity against methicillin-resistantStaphylococcus aureus(MRSA).

With the rise in microbial resistance to traditional antibiotics and disinfectants, there is a pressing need for the development of novel and effective antibacterial agents. Two major approaches being adopted worldwide to overcome antimicrobial resistance are the use of plant leaf extracts and metallic nanoparticles (NPs). However, there are no reports on the antibacterial potential of NPs coated with plant extracts, which may lead to novel ways of treating infections. This study presents an innovative approach to engineer antibacterial NPs by leveraging the inherent antibacterial properties of zinc oxide NPs (ZnO NPs) in combination withAzadirachta indica(AI) leaf extract, resulting in enhanced antibacterial efficacy. ZnO NPs were synthesised by the precipitation method and subsequently coated withAIleaf extract to produce ZnO-AInanocore-shell structures. The structural and morphological characteristics of the bare and leaf extract coated ZnO NPs were analysed by x-ray diffraction and field emission scanning electron microscopy, respectively. The presence of anAIleaf extract coating on ZnO NPs and subsequent formation of ZnO-AInanocore-shell structures was verified through Fourier transform infrared spectroscopy and photoluminescence techniques. The antibacterial efficacy of both ZnO NPs and ZnO-AInanocore-shell particles was evaluated against methicillin-resistantStaphylococcus aureususing a zone of inhibition assay. The results showed an NP concentration-dependent increase in the diameter of the inhibition zone, with ZnO-AInanocore-shell particles exhibiting superior antibacterial properties, owing to the combined effect of ZnO NPs and the poly phenols present inAIleaf extract. These findings suggest that ZnO-AInanocore-shell structures hold promise for the development of novel antibacterial creams and hydrogels for various biomedical applications.

Development of a yeast cell based method for efficient screening of high yield tacrolimus production strain.

Tacrolimus (FK506) is a widely used and clinically important immunosuppressant drug that can be produced by fermentation of Streptomyces tsukubaensis. The industrial strains are typically obtained through multiple rounds of mutagenesis and screening, a labor-intensive process. We have established an efficient yeast cell based screening method for the evolutionary process of high-FK506-yielding strain. The S. tsukubaensis strains of different FK506 yields were tested for zone of growth inhibition of the wild type and calcineurin mutant (cnb1∆) yeast strains. We found that different FK506 yields correspond well to altered yeast growth inhibitions. Based on the combinational inhibition effects of FK506 with different antifungals that have been frequently reported, we also tested the zone of inhibition by addition of fluconazole, amphotericin B and caspofungin to the medium. In the end, for the best screening performance, we systemically evaluated the strategy when different yeast strains and different antifungals were used according to the clarity, size, and divergence of the inhibition circles. Using different yeast strains and antifungals, we successfully broadened the screening spectrum. An efficient high-FK506-yield S. tsukubaensis screening method has been established and optimized. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03870-y.

Enhancement of antimicrobial properties by metals doping in nano-crystalline hydroxyapatite for efficient biomedical applications.

In this study, we have introduced a method for the synthesis of various metal-doped nano-crystalline hydroxyapatites (HAp) using a standard wet chemical precipitation technique. Both divalent (Ni and Zn) and trivalent (Al and Fe) metals were selected for the doping process. Additional research work was also conducted to assess the antimicrobial efficacy of these doped-HAps against a range of gram-positive and gram-negative microorganisms. All the synthesized metal-doped hydroxyapatite (HAp) exhibited notable antibacterial characteristics against gram-negative bacterial strains, namely Escherichia coli (E. coli) and Salmonella typhi (S. typhi), outperforming the pure HAp. The inhibition zone observed for the metal-doped HAp ranged from 14 to 16 mm. The Fe ion displayed a notable inhibitory zone measuring 16 mm, proving to be the most expansive among all tested ions against both E. coli and S. typhi bacterial strains. The Zn-HAp exhibited a comparable inhibitory zone size of 14 mm against both S. typhi and E. coli. Additional characterization methods, such as X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, and Scanning electron microscopy (SEM), were used to validate the structural properties of the synthesized metal-doped hydroxyapatite (HAp) samples. The biocompatibility assessment of metal-doped hydroxyapatite (HAp) samples was carried out by haemolysis tests, which revealed that all synthesized hydroxyapatite (HAp) samples have the potential to serve as reliable biomaterials.

An antimicrobial activity of oil extracted from Saara hardwickii / Atividade antimicrobiana do óleo extraído de Saara hardwickii

Present research work represents antiviral and antibacterial value of body fat of Saara hardwickii commonly called as spiny tailed lizard. Oil was extracted from body fats located in the ventral region of this animal using hydrocarbons e.g., n-hexane, methanol, butanol and ethyl acetate as a solvent. The antibacterial activity of lizard oil was tested against standard as well as multi-resistant lines ofEscherichia coli, Styphalococcus aureus, Pseudomonas aeruginosa and Proteus vulgaris alone and with antibiotic ampicillin. For antibacterial potential, Ethyl acetate and Butanol solvent extract showed best zone of inhibition (7mm) with P. aeruginosa and S. aureus respectively. For antiviral potential, Butanol and Methanol extract showed best HA (Hemagglutination) titer of 04 with NDV and IBV viral strain respectively. It is concluded that lizard oil has antimicrobial potential against different pathogens strains (virus, bacteria).

O presente trabalho de pesquisa apresenta a importância antiviral e antibacteriana da gordura corporal de Saara hardwickii, comumente chamado de lagarto de cauda espinhosa. O óleo foi extraído de gorduras corporais localizadas na região ventral desse animal usando hidrocarbonetos, por exemplo, n-hexano, metanol, butanol e acetato de etila, como solvente. A atividade antibacteriana do óleo do lagarto foi testada em linhagens padrão e multirresistentes de Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa e Proteus vulgaris, de forma isolada e com antibiótico ampicilina. Para o potencial antibacteriano, acetato de etila e extrato de butanol apresentaram melhor zona de inibição (7 mm) com P. aeruginosa e S. aureus, respectivamente. Para o potencial antiviral, o extrato de butanol e o extrato de metanol apresentaram melhor título de hemaglutinação de 4 com as cepas virais NDV e IBV, respectivamente. Conclui-se que o óleo do lagarto possui potencial antimicrobiano contra diferentes cepas de patógenos (vírus e bactérias).

An antimicrobial activity of oil extracted from Saara hardwickii

Abstract Present research work represents antiviral and antibacterial value of body fat of Saara hardwickii commonly called as spiny tailed lizard. Oil was extracted from body fats located in the ventral region of this animal using hydrocarbons e.g., n-hexane, methanol, butanol and ethyl acetate as a solvent. The antibacterial activity of lizard oil was tested against standard as well as multi-resistant lines ofEscherichia coli, Styphalococcus aureus, Pseudomonas aeruginosa and Proteus vulgaris alone and with antibiotic ampicillin. For antibacterial potential, Ethyl acetate and Butanol solvent extract showed best zone of inhibition (7mm) with P. aeruginosa and S. aureus respectively. For antiviral potential, Butanol and Methanol extract showed best HA (Hemagglutination) titer of 04 with NDV and IBV viral strain respectively. It is concluded that lizard oil has antimicrobial potential against different pathogens strains (virus, bacteria).

Resumo O presente trabalho de pesquisa apresenta a importância antiviral e antibacteriana da gordura corporal de Saara hardwickii, comumente chamado de lagarto de cauda espinhosa. O óleo foi extraído de gorduras corporais localizadas na região ventral desse animal usando hidrocarbonetos, por exemplo, n-hexano, metanol, butanol e acetato de etila, como solvente. A atividade antibacteriana do óleo do lagarto foi testada em linhagens padrão e multirresistentes de Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa e Proteus vulgaris, de forma isolada e com antibiótico ampicilina. Para o potencial antibacteriano, acetato de etila e extrato de butanol apresentaram melhor zona de inibição (7 mm) com P. aeruginosa e S. aureus, respectivamente. Para o potencial antiviral, o extrato de butanol e o extrato de metanol apresentaram melhor título de hemaglutinação de 4 com as cepas virais NDV e IBV, respectivamente. Conclui-se que o óleo do lagarto possui potencial antimicrobiano contra diferentes cepas de patógenos (vírus e bactérias).

Antagonistic activity of Bacillus amyloliquefaciens subsp. amyloliquefaciens against multidrug resistant Serratia rubidaea.

Serratia rubidaea is an opportunistic Gram-negative pathogen that has developed antimicrobial resistance to a variety of commercial antibiotics. The spread of this multidrug-resistant pattern predicts that it will get harder and harder to treat S. rubidaea infections in the future. For this perception, antimicrobial proteins might represent a safe, effective, and biodegradable alternative because their site of action is on cyclic peptides. In this study, one candidate Bacillus amyloliquefaciens subsp. amyloliquefaciens was isolated from the soil of Sundarban mangrove forest, and its identification was confirmed both using the PCR (Polymerase chain reaction) method and the BIOLOG™ microbial identification system. The antibacterial protein, which has a molecular mass of about 50 kDa, was isolated from B. amyloliquefaciens subsp. amyloliquefaciens. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to confirm the extracted protein's purity. This potential protein was discovered to develop and exhibit antagonistic activity throughout a broad temperature, pH, and salinity range. At doses ranging from 300 to 400 µg/ml, this protein has antagonistic activity against multidrug resistant S. rubidaea and a wide range of other resistant pathogenic bacteria such as Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and so on. The research provides new insights to develop bio-control agents that can be applied for prevent, treat, and control infectious diseases caused by multidrug resistant S. rubidaea, as well as other pathogenic bacteria.

Investigating the Molecular Interactions of Quinoline Derivatives for Antibacterial Activity Against Bacillus subtilis: Computational Biology and In Vitro Study Interpretations.

Bacterial infections are evolving and one of the chief problems is emergence and prevalence of antibacterial resistance. Moreover, certain strains of Bacillus subtilis have become resistant to several antibiotics. To counteract this menace, the present work aimed to comprehend the antibacterial activity of synthesized two quinoline derivatives against Bacillus subtilis. Toxicity predictions via Protox II, SwissADME and T.E.S.T (Toxicity Estimation Software Tool) revealed that these derivatives were non-toxic and had little to no adverse effects. Molecular docking studies carried out in Schrodinger with two quinoline derivatives (referred Q1 and Q2) docked against selected target proteins (PDB IDs: 2VAM and1FSE) of B. subtilis demonstrated ideal binding energies (2VAM-Q1: - 4.63 kcal/mol and 2VAM-Q2: - 4.46 kcal/mol, and 1FSE-Q1: - 3.51 kcal/mol, 1FSE-Q2: - 6.34 kcal/mol). These complexes were simulated at 100 ns and the outcomes revealed their stability with slight conformational changes. Anti-microbial assay via disc diffusion method revealed zones of inhibition showing that B. subtilis was inhibited by both Q1 and Q2, with Q2 performing slightly better than Q1, pointing towards its effectiveness against this organism and necessitating further study on other bacteria in prospective studies. Thus, this study demonstrates that our novel quinoline derivatives exhibit antibacterial properties against Bacillus subtilis and can act as potent anti-bacterials.

Antimicrobial Efficacy of Three Different Mouthwashes on Periodontal Pathogens-An In vitro Study.

Aim: The aim of this research was to identify the antimicrobial effectiveness of three different mouthwashes on periodontal pathogenic microorganisms. Materials and Methods: 2 periodontal disease-causing microorganisms, i.e., Aggregatibacter actinomycetemcomitans, and Porphyromonas gingivalis, were chosen for this investigation. Prior to commencing this research, a variety of branded and commercially obtainable mouthwashes were procured. Three oral rinses, namely HiOra, Hexidine, and Amflor, were chosen for the current research. The subculture of A. actinomycetemcomitans as well as P. gingivalis was performed by subjecting them to incubation for 48 to 72 hours at 35-37°C. The disk diffusion method was employed to evaluate the antibacterial efficiency of the extract in opposition to the pathogens tested. The zone of inhibition was calculated in millimeters. The mean value of every sample was documented. Results: Hexidine oral rinse in pursuit by Amflor as well as HiOra oral rinse exhibited the highest zone of inhibition in opposition to A. Actinomycetemcomitans and P. gingivalis. The differences amid the groups were statistically significant with a P value < 0.001. Conclusion: The current research concluded that amid the three different oral rinses employed in the current research, Hexidine oral rinse exhibited greatest antimicrobial effectiveness versus Amflor and HiOra mouthrinse.

Assessment of Antimicrobial Activity of Lycopene, Vitamin E, and Lycopene-Vitamin E Combination Against Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis, and Candida albicans: An In Vitro Study.

Background Lycopene is a naturally occurring compound classified as a carotenoid, a group of pigments responsible for the vibrant colors observed in many fruits and vegetables. It is most commonly associated with red-colored fruits and vegetables, such as tomatoes, watermelon, pink grapefruit, and papaya. Vitamin E encompasses a group of chemical compounds that share a structural relationship with alpha-tocopherol and are essential for the proper functioning of the human body. It is a fat-soluble vitamin and is known for its antioxidant properties. The aim of this study is to evaluate the antimicrobial activity of lycopene extract, vitamin E extract, and their combination against oral pathogens for their potential application in the treatment of oral diseases. Materials and methods The potential antimicrobial effects of extracts derived from lycopene, vitamin E, and their combination were evaluated against oral commensals like Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis, and Candida albicans. Three concentrations (25 µl, 50 µl, and 100 µl) of the extract were tested. Mueller-Hinton agar (MHA) and Rose Bengal agar (RBA) bases were utilized to determine the zone of inhibition. And the experiments were repeated in triplicate for each group. Results The identification and assessment of the antimicrobial activity of lycopene extract, vitamin E extract, and their combination revealed the greatest efficacy at the highest concentration (100 µl) against all tested microbial strains. Notably, C. albicans exhibited the highest susceptibility compared to the other strains. Vitamin E had the least antimicrobial effect and combination had the highest antimicrobial effect. Conclusion The results of our study demonstrated substantial antimicrobial activity of lycopene and vitamin E. These findings suggest that lycopene and vitamin E can be harnessed in the development of diverse drug formulations for the treatment of oral diseases.

Evaluation of in vitro activity of herbal formula Di Er You and herb Coptis against bacteria isolated from dogs with otitis externa.

OBJECTIVE: To determine the in vitro activity of the herbal formula Di Er You (DEY) and the single-herb Coptis against bacteria cultured from dogs with otitis externa. ANIMALS: 32 client-owned dogs diagnosed with otitis externa. METHODS: A sample of otic debris from each patient was collected and plated onto a fresh Sheep's Blood Agar plate in the hospital. After bacterial growth was confirmed, 4 wells were created, numbered randomly, and treated with saline (placebo), DEY, Coptis, and Zymox Otic Enzymatic Solution with 1% Hydrocortisone (Zymox). After 24 hours of incubation, the diameter of the zone of inhibition (dZOI) of each treatment was measured and recorded, and compared among treatments. A sample of the bacterial colonies grown was submitted to an outside lab for bacterial identification. RESULTS: The mean ± SD dZOI values for saline, DEY, Coptis, and Zymox treated wells were 0.25 ± 1.41, 12.47 ± 3.92, 14.25 ± 7.12, and 3.22 ± 5.12, respectively. Post hoc multiple comparisons test revealed that (1) saline-treated wells had significantly smaller dZOI values than the other 3 groups (all P < .001), (2) Zymox treated wells had significantly smaller dZOI values than either herbal treated groups (both P < .001), and (3) DEY treated wells had significantly smaller dZOI values than those treated with Coptis (P = .0042). CLINICAL RELEVANCE: The results from this in vitro study suggested that both DEY and Coptis could be effective treatments in inhibiting the growth of bacteria in dogs with otitis externa. Prospective randomized controlled clinical trials are warranted to confirm these findings.

Evaluation of Antibacterial Efficacy of Two Commercially Available Probiotics as Intracanal Medicament against Enterococcus faecalis: An In Vitro Study.

AIM: This study was performed to evaluate the antibacterial efficacy of two commercially available probiotics (BIFILAC and VSL 3) as intracanal medicament against Enterococcus faecalis in endodontic therapy. MATERIALS AND METHODS: Microorganisms from commercially available probiotics (BIFILAC and VSL 3) were extracted via the manufacturer's recommendations and mixed by weight. About 30 microliters were then placed on sterile discs. The pathogenic test organism was E. faecalis set to a 1 McFarland standard challenge. A two-probiotic disc template on blood agar plates was inoculated with E. faecalis and incubated at 37°C for 48 hours and 1 week respectively. Phase-1 of the study was conducted by a disc diffusion assay test to evaluate zones of inhibition (ZOI) in millimeters (mm). Phase-2 was conducted by mixing 9 mL of 30% poloxamer 407 and MRS broth in a test tube, together with the two probiotic mixtures and E. faecalis, set at a 2 McFarland standard. Serial dilutions up to 108 were done and the mixture was placed inside root canals and incubated at 37ºC for 36 hours and evaluated for colony-forming unit (CFU)/mL counts. RESULTS: The results of phase-1 showed that probiotics Lactobacillus rhamnosus and Bifidobacterium species are effective in fighting against E. faecalis with the acceptable zone of inhibition. The results of phase-2 showed that both the probiotics are effective against E. faecalis with a reduction in the number of CFU after probiotic usage. CONCLUSION: Commercially available probiotics can be used effectively as an intracanal medicament to fight against E. faecalis, Poloxamer 407 is a promising vehicle for delivering probiotics inside the root canal system. Further in vitro and in vivo studies are needed to determine the full potential of "Bacteriotherapy" with an application of probiotics. CLINICAL SIGNIFICANCE: If probiotics are proved to be an effective intracanal medicament against E.faecalis they can be used as an alternative to calcium hydroxide as intracanal medicament with no side effects to the host.

Tandem LC-MS Identification of Antitubercular Compounds in Zones of Growth Inhibition Produced by South African Filamentous Actinobacteria.

Novel antitubercular compounds are urgently needed to combat drug-resistant Mycobacterium tuberculosis (Mtb). Filamentous actinobacteria have historically been an excellent source of antitubercular drugs. Despite this, drug discovery from these microorganisms has fallen out of favour due to the continual rediscovery of known compounds. To increase the chance of discovering novel antibiotics, biodiverse and rare strains should be prioritised. Subsequently, active samples need to be dereplicated as early as possible to focus efforts on truly novel compounds. In this study, 42 South African filamentous actinobacteria were screened for antimycobacterial activity using the agar overlay method against the Mtb indicator Mycolicibacterium aurum under six different nutrient growth conditions. Known compounds were subsequently identified through extraction and high-resolution mass spectrometric analysis of the zones of growth inhibition produced by active strains. This allowed the dereplication of 15 hits from six strains that were found to be producing puromycin, actinomycin D and valinomycin. The remaining active strains were grown in liquid cultures, extracted and submitted for screening against Mtb in vitro. Actinomadura napierensis B60T was the most active sample and was selected for bioassay-guided purification. This resulted in the identification of tetromadurin, a known compound, but which we show for the first time to have potent antitubercular activity, with the MIC90s within the range of 73.7-151.6 nM against M. tuberculosis H37RvTin vitro under different test conditions. This shows that South African actinobacteria are a good source of novel antitubercular compounds and warrant further screening. It is also revealed that active hits can be dereplicated by HPLC-MS/MS analysis of the zones of growth inhibition produced by the agar overlay technique.

Antibacterial and Antibiofilm Activities of ß-Lapachone by Modulating the Catalase Enzyme.

BACKGROUND: Bacterial infections constantly have a large impact on public health, because of increased rates of resistance and reduced frequency of development of novel antibiotics. The utility of conventional antibiotics for treating bacterial infections has become increasingly challenging. The aim of the study was to assess the antibacterial effect of ß-Lapachone (ß-Lap), a novel synthetic compound. METHODS: The antibacterial activity of the ß-Lap compound was examined against laboratory strains by agar well diffusion method and broth dilution assay. Growth kinetics in presence of ß-Lap on Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa (ATCC 27853) were assessed by microplate alamarBlue assay. Crystal violet blue assay was used for biofilm inhibition and biofilm eradication. P. aeruginosa catalase (KatA) complexed with ß-Lap was modeled using molecular docking approach. RESULTS: ß-Lap exhibited potent antimicrobial activity against laboratory strains of bacteria with MIC of 0.2 mM for S. saprophyticus and Staphylococcus aureus, and 0.04 mM for Staphylococcus epidermidis and Pseudomonas aeruginosa ATCC 27853. The inhibition of catalase enzyme was found to be the cause for its antibacterial activity. Bioinformatics analysis suggests that ß-Lap can inhibit KatA activity by interacting with catalase proximal active site and heme binding site. The activity of some commercial antibiotics was enhanced in association with ß-Lap. In addition, ß-Lap inhibited the biofilm formation and eradicated the already formed and ultra-mature biofilms of aforesaid bacterial strains. CONCLUSION: These observations indicated that ß-Lap could be a promising antibacterial agent for the treatment and prevention of infectious diseases.

The quality by design approach for optimization of slayer exciter based low power portable atmospheric plasma jet on bactericidal efficacy of Pseudomonas aeruginosa.

A simple, portable, economical low-temperature atmospheric plasma (LTAP) for bactericidal efficacy of Gram-negative bacteria (Pseudomonas aeruginosa) with different carrier gases (argon, helium, and nitrogen) using the quality by design (QbD) approach, design of experiments (DoE), and response surface graphs (RSG) is presented. Box-Behnken design was used as the DoE to narrow down and further optimize the experimental factors of LTAP. Plasma exposure time, input DC voltage, and carrier gas flow rate were varied to examine the bactericidal efficacy using the zone of inhibition (ZOI). A higher bactericidal efficacy was achieved under the optimal bactericidal factors having ZOI of 50.837 ± 2.418 mm2 with the plasma power density of 132 mW/cm3 for LTAP-Ar at 61.19 s, 14.8747 V, and 219.379 sccm than LTAP-He and LTAP-N2 . The LTAP-Ar was further evaluated at different frequencies and probe lengths to achieve a ZOI of 58.237 ± 4.01 mm2 .

A zone-of-inhibition assay to screen for humoral antimicrobial activity in mosquito hemolymph.

In insects, antibacterial immunity largely depends on the activation of downstream signaling and effector responses, leading to the synthesis and secretion of soluble effector molecules, such as antimicrobial peptides (AMPs). AMPs are acute infection response peptides secreted into the hemolymph upon bacterial stimulation. The transcription of innate immunity genes encoding for AMPs is highly dependent on several signaling cascade pathways, such as the Toll pathway. In the African malaria mosquito, Anopheles gambiae, AMPs hold a special interest as their upregulation have been shown to limit the growth of malaria parasites, bacteria, and fungi. Most of the current knowledge on the regulation of insect AMPs in microbial infection have been obtained from Drosophila. However, largely due to the lack of convenient assays, the regulation of antimicrobial activity in mosquito hemolymph is still not completely understood. In this study, we report a zone of inhibition assay to identify the contribution of AMPs and components of the Toll pathway to the antimicrobial activity of A. gambiae hemolymph. As a proof of principle, we demonstrate that Micrococcus luteus challenge induces antimicrobial activity in the adult female mosquito hemolymph, which is largely dependent on defensin 1. Moreover, by using RNAi to silence Cactus, REL1, and MyD88, we showed that Cactus kd induces antimicrobial activity in the mosquito hemolymph, whereas the antimicrobial activity in REL1 kd and MyD88 kd is reduced after challenge. Finally, while injection itself is not sufficient to induce antimicrobial activity, our results show that it primes the response to bacterial challenge. Our study provides information that increases our knowledge of the regulation of antimicrobial activity in response to microbial infections in mosquitoes. Furthermore, this assay represents an ex vivo medium throughput assay that can be used to determine the upstream regulatory elements of antimicrobial activity in A. gambiae hemolymph.