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

AIM: The present study aimed to evaluate the antimicrobial efficacy of silver nanoparticles infused with Azadirachta indica extract and chlorhexidine against red-complex periopathogens. MATERIALS AND METHODS: 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. RESULTS: 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). CONCLUSION: 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. CLINICAL SIGNIFICANCE: 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.

Standardisation of high throughput microdilution antifungal susceptibility testing for Candida albicans and Cryptococcus neoformans.

The Clinical and Laboratory Standards Institute (CLSI) M27 guidelines are the recommended and most commonly used protocols for broth microdilution antifungal susceptibility testing of yeasts. However, these guidelines are limited to the use of 96-well assay plates, limiting assay capacity. With the increased risk of fungal resistance emerging in the community, it is important to have alternative protocols available, that offer higher throughput and can screen more than eight to ten potential antifungal compounds per plate. This study presents an optimised broth microdilution minimum inhibitory concentration (MIC) method for testing the susceptibility of yeasts in an efficient high throughput screening setup, with minimal growth variability and maximum reproducibility. We extend the M27 guidelines and optimise the conditions for 384-well plates. Validation of the assay was performed with ten clinically used antifungals (fluconazole, amphotericin B, 5-fluorocytosine, posaconazole, voriconazole, ketoconazole, itraconazole, caspofungin diacetate, anidulafungin and micafungin) against Candida albicans and Cryptococcus neoformans.

Comparative Evaluation of Antimicrobial Activity and Minimum Inhibitory Concentration of Commercially Available Pediatric Dentifrices: An In Vitro Study.

Aim: The aim of this study was to evaluate the antimicrobial efficacy and minimum inhibitory concentration (MIC) of commercially available pediatric dentifrices containing different compositions against Streptococcus mutans and Lactobacillus activity. Materials and methods: Four different commercially available brands of pediatric dentifrices, designated as sample I-fluoride, sample II-herbal, sample III-xylitol with nanosilver particles, and sample IV-xylitol with fluoride, along with two control groups (a positive control-ciprofloxacin and a negative control-distilled water), were tested for their antibacterial activity by measuring the zone of inhibition, followed by MIC against two dental bacterial pathogens, S. mutans strain and Lactobacillus acidophilus (LB) strain, at five different twofold dilutions of 100, 50, 25%, 12.5, and 6.25% concentrations. Result: All four dentifrices were found to have wide variations in their effectiveness against the two tested microorganisms at 100% (pure) and 50% concentrations, with sample I having the highest activity, followed by sample IV and sample II. At 25% concentration, only sample I and sample IV showed antibacterial activity, while at 12.5 and 6.25% concentrations, none of the tested toothpastes exhibited any antibacterial activity. Sample III failed to show antibacterial activity even in pure form against the two microorganisms. Conclusion: In our present study, the fluoride-containing pediatric dentifrice with a lower fluoride concentration (458 ppm) exhibited the highest zone of inhibition, followed by the xylitol with fluoride dentifrice and the herbal dentifrice. No zone of inhibition was observed in the nanosilver with xylitol dentifrice. How to cite this article: Dureha R, Navit S, Khan SA, et al. Comparative Evaluation of Antimicrobial Activity and Minimum Inhibitory Concentration of Commercially Available Pediatric Dentifrices: An In Vitro Study. Int J Clin Pediatr Dent 2024;17(8):938-944.

Use of Cyrene™, as an alternative to dimethyl sulfoxide, as a diluent for Melatonin to determine its in vitro antimicrobial capacity.

Melatonin (MLT) is a methoxyindole that has potent antioxidant actions, anti-inflammatory, and antiapoptotic capacity. However, its in vitro antibacterial capacity has been the least studied of its properties. Dimethylsulfoxide (DMSO) has been the most used solvent for these tests, but it shows an antimicrobial effect if it is not dissolved. Cyrene™ is a new solvent that has emerged as an alternative to DMSO. Therefore, this study aimed to determine the antimicrobial capacity of MLT by MIC assays, using Cyrene™ as a solvent. Likewise, the solubility of MLT in this solvent and whether it exerted any effect on bacterial growth at different percentages was also determined. Different dilutions of MLT in Cyrene™ with different concentrations, were prepared. No growth inhibition caused by MLT was observed. The growth inhibition observed was because of Cyrene™. The maximum amount of MLT that can be diluted in 100% Cyrene is 10 mg/mL, but this percentage of solvent shows a bactericidal effect. Therefore, it must be dissolved at 5% to avoid this effect, so only 4 mg/mL of MLT can be diluted in it. Therefore, if no other solvents are available, the in vitro antibacterial role of MLT cannot be adequately assessed.

Post-Antibiotic and Post-Antibiotic Sub-Minimum Inhibitory Concentration Effects of Carvacrol against Salmonella Typhimurium.

Carvacrol is a compound present in essential oils with proven antimicrobial activity against numerous pathogens. We firstly determine the post-antibiotic effect (PAE) of carvacrol (1×, 2×, 4× MIC) and post-antibiotic sub-minimum inhibitory concentration (MIC) effect (1× + 0.25× MIC and 2× + 0.25× MIC) for two concentrations of Salmonella Typhimurium ATCC14028 (106 and 108 CFU/mL). Prior to testing, the minimum concentration and exposure time to achieve the bacterial inhibition (MIC 0.6 mg/mL and 10 min) were determined by broth microdilution and time-kill curve methods, respectively. At the MIC, carvacrol did not generate any PAE. At twice the MIC, the PAE was 2 h with the standard inoculum (106 CFU/mL) and 1 h with the high-density inoculum (108 CFU/mL). At 4× MIC concentrations, the PAE was higher in both cases > 43.5 h. Continuous exposure of post-antibiotic phase bacteria (1× and 2× MIC) to carvacrol at 0.25× MIC (0.15 mg/mL) resulted in an increase in PAE (PA-SME) above 43.5 h with both inocula. These results suggest that the PA-SME of carvacrol for S. Typhimurium can be significantly prolonged by increasing the sub-MICs, which would allow dose spacing, reduce adverse effects and improve its efficacy in the treatment of infected animals and as a disinfectant in agri-food facilities.

Assessment of antioxidant and antibacterial efficacy of some indigenous vegetables consumed by the Manipuri community in Sylhet, Bangladesh.

The rapid dietary changes experienced by indigenous people worldwide threaten the use of traditional foods, which are often undervalued. This study focused on evaluating the antioxidant and antibacterial efficacy of five vegetables typically consumed by the Manipuri ethnic groups in the Sylhet region of Bangladesh: Yongchak seed (Parkia speciosa), Telikadam seed (Leucaena leucocephala), Phakphai leaf (Persicaria odorata), Sheuli leaf (Nyctanthes arbor-tristis), and bamboo shoot (Bambusa spp.). The samples were dried and powdered to assess the antioxidant activity through total phenolic content (TPC), total flavonoid content (TFC), total tannin content (TTC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. Antibacterial efficacy was determined by measuring the zone of inhibition (ZOI), minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC). Leafy vegetables exhibited higher TPC, TFC, and TTC than seeds and shoots, with N. arbor-tristis leaf showing the highest TPC (99.16 ± 2.07 mg GAE/g DW) and P. odorata leaf exhibiting the highest TFC (9.19 ± 0.7 mg QE/g) and TTC (3.59 ± 0.26 mg TAE/g). However, Bambusa spp. shoot extract showed the highest antioxidant potential (IC50: 1.66 ± 0.05 mg/mL). All samples exhibited higher ZOI against gram-positive bacteria (Bacillus spp. and Staphylococcus spp.), ranging from 10 ± 2.65 to 19.33 ± 2.08 mm. L. leucocephala seed extract showed the highest antibacterial activity against both the tested gram-positive bacteria with a MIC of 15.6 mg/mL. Conversely, the P. odorata leaf extract exerted the strongest antibacterial effect against gram-negative bacteria, with the lowest MIC values for Klebsiella spp. (31.25 mg/mL) and Escheria coli (62.5 mg/mL). The findings of this investigation suggest that the selected indigenous vegetables could be valuable sources of phytochemicals with potential antioxidant and antibacterial activities. Incorporating and promoting these traditional foods into the diet may improve food security, dietary diversity, and public health in Bangladesh.

A Comprehensive Review of Silver and Gold Nanoparticles as Effective Antibacterial Agents.

The increasing threat from antibiotic-resistant bacteria has necessitated the development of novel methods to counter bacterial infections. In this context, the application of metallic nanoparticles (NPs), especially gold (Au) and silver (Ag), has emerged as a promising strategy due to their remarkable antibacterial properties. This review examines research published between 2006 and 2023, focusing on leading journals in nanotechnology, materials science, and biomedical research. The primary applications explored are the efficacy of Ag and Au NPs as antibacterial agents, their synthesis methods, morphological properties, and mechanisms of action. An extensive review of the literature on NPs synthesis, morphology, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and effectiveness against various Gram(+/-) bacteria confirms the antibacterial efficacy of Au and Ag NPs. The synthesis methods and characteristics of NPs, such as size, shape, and surface charge, are crucial in determining their antibacterial activity, as these factors influence their interactions with bacterial cells. Furthermore, this review underscores the urgent necessity of standardizing synthesis techniques, MICs, and reporting protocols to enhance the comparability and reproducibility of future studies. Standardization is essential for ensuring the reliability of research findings and accelerating the clinical application of NP-based antimicrobial approaches. This review aims to propel NP-based antimicrobial strategies by elucidating the properties that enhance the antibacterial activity of Ag and Au NPs. By highlighting their inhibitory effects against various bacterial strains and relatively low cytotoxicity, this work positions Ag and Au NPs as promising materials for developing antibacterial agents, making a significant contribution to global efforts to combat antibiotic-resistant pathogens.

In vitro and in vivo efficacy of zinc oxide green nanoparticles against multidrug-resistant Salmonella Typhi.

Antibiotic resistance is an increasing threat, requiring novel therapeutic solutions. Metal nanoparticles e.g., zinc oxide nanoparticles (ZnO NPs) exhibited the potential against many bacterial pathogens. Strains of Salmonella enterica serovar Typhi resistant to ceftriaxone were reported first from Pakistan in 2016. Since then, S. Typhi is a pathogen of concern globally owing to its rapidly emerging resistance potential against many last resort antibiotics. In the present study, in vitro and in vivo antimicrobial activity of ZnO NPs against multidrug resistant (MDR) and extensively drug resistant (XDR) Salmonella Typhi strains from Pakistan was evaluated. Zinc oxide green nanoparticles (ZnO GNPs), synthesized from Aloe vera, were characterized by SEM, XRD, UV-vis and Raman spectroscopy. In vitro antibacterial activity of two different concentrations of ZnO GNPs (7 and 15%) was checked using agar well diffusion method. Further, broth microdilution and time kill assays were performed using the ZnO GNPs. In vivo assays were conducted in BALB/c mice sepsis models. In all the three methods, agar well diffusion assay broth microdilution and time kill assay, different zinc oxide dihydrate precursor concentrations had shown the antibacterial activity. The minimum inhibitory concentration (MIC) of ZnO GNPs nanoparticles against MDR and XDR S. Typhi strains was found as 16 to 64 µg/ml. In vivo experiment has shown a significant decrease in CFU/ml in the mice treated with ZnO GNPs as compared to the control group. Our findings have revealed that ZnO GNPs have significant antibacterial activity against MDR and XDR S. Typhi, both in vitro and in vivo.

Evaluation of the cytotoxicity and antibacterial activity of nano-selenium prepared via gamma irradiation against cancer cell lines and bacterial species.

A recent scientific investigation has shown promising results of selenium nanoparticles (SeNPs) for the anticancer and antimicrobial activities. This study aims to evaluate the effects of PVP SeNPs on bacterial strains, including Staphylococcus aureus (S. aureus), Bacillus cereus (B. cereus), Klebsiella pneumoniae (K. pneumoniae), Escherichia coli (E. coli), and Pseudomonas aeruginosa (P. aeruginosa). Also, its antitumor activity against the MRC-5 carcinoma cell line. SeNPs were prepared via gamma irradiation using PVP as a capping agent, and their size and morphological structure were determined using HRTEM. The size of the SeNPs ranged from 36 to 66.59 nm. UV-vis spectra confirmed the formation of SeNPs, while FTIR measurement confirmed a change in the PVP structure after adding selenium nanoparticles. The highest effect was reported on HepG2 by an IC50 with a value of 8.87 µg/ml, followed by HeLa, PC3, MCF-7, and Caco2 cell lines, respectively. Furthermore, ZOI reached 36.33 ± 3.05 mm. The best value of the minimum inhibitory concentration (MIC) was 0.313 µg/ml. Scanning electron microscope (SEM) imaging against bacteria showed deformations and distortions in their structures. Transmission electron (TEM) revealed ultrastructure changes in treated bacteria because of the free radicals that made cytotoxicity which confirmed by Electron spin resonance (ESR).

Association between in vitro susceptibility and clinical outcomes in fungal keratitis.

PURPOSE: The purpose of this study was to assess the association between antifungal susceptibility as measured by minimum inhibitory concentration (MIC) and clinical outcomes in fungal keratitis. METHODS: This pre-specified secondary analysis of the Mycotic Ulcer Treatment Trial II (MUTT II) involved patients with filamentous fungal keratitis presenting to Aravind Eye Hospitals in South India. Antifungal susceptibility testing for natamycin and voriconazole was performed on all samples with positive fungal culture results according to Clinical and Laboratory Standards Institute Guidelines. The relationship between MIC and clinical outcomes of best-corrected visual acuity, infiltrate or scar size, corneal perforation, need for therapeutic penetrating keratoplasty, and time to re-epithelialization were assessed. RESULTS: We obtained MIC values from 141 patients with fungal keratitis. The most commonly cultured organisms were Aspergillus (46.81%, n = 66) and Fusarium (44.68%, n = 63) species. Overall, there was no association between antifungal MICs and clinical outcomes. Subgroup analysis revealed that among Fusarium-positive cases, higher voriconazole MIC was correlated with worse three-month best-corrected visual acuity (p = 0.03), increased need for therapeutic penetrating keratoplasty (p = 0.04), and time to re-epithelialization (p = 0.03). No significant correlations were found among Aspergillus-positive cases. There were no significant correlations found between natamycin MIC and clinical outcomes among organism subgroups. CONCLUSIONS: Decreased susceptibility to voriconazole was associated with increased odds of requiring a therapeutic penetrating keratoplasty in Fusarium-positive cases. Susceptibility to natamycin was not associated with any of the measured outcomes.

Antimicrobial and Phylogenomic Characterization of Bacillus cereus Group Strains Isolated from Different Food Sources in Italy.

Background:Bacillus cereus is a widespread environmental Gram-positive bacterium which is especially common in soil and dust. It produces two types of toxins that cause vomiting and diarrhea. At present, foodborne outbreaks due to Bacillus cereus group bacteria (especially Bacillus cereus sensu stricto) are rising, representing a serious problem in the agri-food supply chain. Methods: In this work, we analyzed 118 strains belonging to the Bacillus cereus group, isolated from several food sources, for which in vitro and in silico antibiotic resistance assessments were performed. Results: Many strains showed intermediate susceptibility to clindamycin, erythromycin, and tetracycline, suggesting an evolving acquisition of resistance against these antibiotics. Moreover, one strain showed intermediate resistance to meropenem, an antibiotic currently used to treat infections caused by Bacillus cereus. In addition to the phenotypic antimicrobial resistance profile, all strains were screened for the presence/absence of antimicrobial genes via whole-genome sequencing. There was inconsistency between the in vitro and in silico analyses, such as in the case of vancomycin, for which different isolates harbored resistance genes but, phenotypically, the same strains were sensitive. Conclusions: This would suggest that antibiotic resistance is a complex phenomenon due to a variety of genetic, epigenetic, and biochemical mechanisms.

Udder health, bacterial isolation and antimicrobial sensitivity of Staphylococcus species from non-dairy goats on smallholder farms in Hong Kong.

This research article describes an investigation into the udder health, bacterial isolation and antimicrobial sensitivity of three staphylococcal species isolated from the milk of non-dairy goats, suckling their kids, on two smallholder farms in the Hong Kong Special Administrative Region. Udder lesions were visually noted in 21 of 34 goats and two goats had palpable abnormalities. Collected milk samples grew a total of 11 bacterial organisms and the most frequently isolated organism was Staphylococcus chromogenes. Selected isolates of S. aureus, caprae and simulans from both farms were tested by antimicrobial sensitivity testing for 23 antimicrobials and all isolates showed antimicrobial resistance to doxycycline and tetracycline. Less common resistance was shown to ampicillin, chloramphenicol, penicillin and rifampicin. This preliminary study confirms the presence of udder lesions and mastitis bacteria in non-dairy goats in Hong Kong, along with the first information on the antimicrobial profile of three common Staphylococcus species bacteria affecting goats.

Antibacterial Activity of Cissus quadrangularis (Veldt Grape) Against Porphyromonas gingivalis, a Keystone Pathogen in Periodontal Disease: An In Vitro Study.

BACKGROUND: Porphyromonas gingivalis, a keystone pathogen and one of the primary pathogens responsible for periodontitis, leads to a chronic inflammatory condition that destroys the periodontal tissues and ultimately results in tooth loss. While conventional non-surgical therapy combined with antibiotics and local drug delivery systems are commonly used to treat periodontitis, certain medicinal herbs have also demonstrated efficacy in its prevention. Cissus quadrangularis L. (CQ), a perennial plant from the Vitaceae family, is widely recognized and used as a medicinal herb in many tropical countries, predominantly in India, Sri Lanka, Thailand, Java, West Africa, and the Philippines. AIM: The aim of the study was to determine the antibacterial activity of CQ against the periodontal keystone pathogen P. gingivalis. METHOD: Aqueous and ethanolic extracts of CQ were prepared using a Soxhlet extractor. The antibacterial effectiveness of these extracts against the periodontal pathogen P. gingivalis was evaluated at different concentrations, and the minimal inhibitory concentration (MIC) was determined using broth microdilution. RESULTS: The ethanolic extract of CQ mixed with 10% dimethyl sulfoxide (DMSO) showed higher inhibition compared to the aqueous extract of CQ against P. gingivalis. CONCLUSION: Our study revealed the potent inhibitory effects of CQ against P. gingivalis. Both aqueous and ethanolic extracts displayed MIC values of 500 µg/mL. Notably, the ethanolic extract of CQ, dissolved in 10% DMSO, demonstrated superior efficacy with a lower IC50 value of 194.36 µg/mL. These findings indicate promising potential for CQ in the management of periodontal disease.

Case of tinea corporis caused by a terbinafine-sensitive Trichophyton indotineae strain in a Vietnamese worker in Japan.

A 42-year-old Vietnamese egg factory worker in Ishikawa prefecture, Japan, presented with itchy concentric erythema on the trunk and left calf. The lesions tested positive by direct potassium hydroxide examination, and two fungal strains were isolated. The isolates produced conidia abundantly and were morphologically indistinguishable from Trichophyton mentagrophytes/interdigitale, but were identified as Trichophyton indotineae by internal transcribed spacer sequence of ribosomal DNA. The lesions were refractory to treatment with topical luliconazole (LLCZ) cream for 4 weeks but subsided with oral itraconazole (ITCZ) 100 mg/day for 4 weeks in combination with topical lanoconazole (LCZ) cream. The lesions recurred 6 weeks after discontinuation of oral ITCZ, and an additional isolate was cultured. The minimum inhibitory concentrations of antimycotics for the isolate cultured at the first visit were: terbinafine (TBF) 0.03 µg/mL, ITCZ 0.015 µg/mL, LLCZ 0.0005 µg/mL, and LCZ 0.002 µg/mL. No TBF-resistant mutation in the amino acid sequence of squalene epoxidase, i.e., Leu 393 Ser/Phe or Phe 397 Leu, was detected in the isolate. The reason for recalcitrance in this case, despite the isolate's sensitivity to antimycotics, was unclear. Possible factors include insufficient use of the antimycotics, incomplete removal of abundantly produced conidia from the lesions, the patient's environment, and a language gap between the patient and physician hindering communication.

Identification and antifungal sensitivity of Fusarium species isolated from piscine hosts.

Fusarium is a huge genus of filamentous fungi that has the potential to cause emerging diseases. Members of this genus can cause infections in plants, animals and humans. Here, we report the isolation of F. oxysporum and F. equiseti from 2 important fish species, Oncorhynchus mykiss (rainbow trout) and Tor putitora (golden mahseer), respectively. F. oxysporum has emerged as a significant fungal pathogen causing infection in many fish. However, F. equiseti has been isolated mainly from plants. As far as the available literatures are concerned, this is the first report on the isolation of F. oxysporum and F. equiseti from these hosts. The isolates were identified based on growth morphology and microscopic observation. F. oxysporum produced violet pigmentation on potato dextrose agar, while F. equiseti had yellow colouration. F. oxysporum produced 1- to 2-celled microconidia along with straight or curved macroconidia having 3 to 4 septa. F. equiseti produced abundant macroconidia with 4 or more septa. Species were further confirmed based on the nucleotide sequences of the internal transcribed spacer region. In a molecular phylogeny analysis, F. oxysporum and F. equiseti formed 2 different clades. In an antifungal sensitivity assay, F. oxysporum was found to be susceptible to clotrimazole with a minimum inhibitory concentration of 1.0 µg ml-1, whereas F. equiseti was susceptible to clotrimazole, ketoconazole and fluconazole. Overall, the main findings of this study are the infection of new hosts by Fusarium species and the limited activity of many antifungal drugs against these pathogens.

Phytochemical profiling and evaluation of antimicrobial activities of common culinary spices: Syzygium aromaticum (clove) and Piper nigrum (black pepper).

Background: The increasing resistance of microbial pathogens to conventional antibiotics necessitates the exploration of alternative antimicrobial agents. This study aims to evaluate the antimicrobial potential and phytochemical properties of Syzygium aromaticum (clove) and Piper nigrum (black pepper) extracts, both of which are known for their historical use in traditional medicine and culinary applications. Methods: Hydroalcoholic and aqueous extracts of clove and black pepper were prepared. The antimicrobial activity of these extracts was assessed using the disk diffusion method against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Aspergillus niger. Minimum inhibitory concentration (MIC) was determined using the broth dilution method. Qualitative phytochemical screening identified the presence of key bioactive compounds, while quantitative analysis measured total phenolic and flavonoid contents. LC-HRMS/MS analysis of ethanolic extracts was performed. Results: Both spices extracts exhibited significant antimicrobial activity, with inhibition zones ranging from 14 to 18 mm. clove showed superior antimicrobial efficacy compared to black paper, particularly against fungi. MIC values ranged between 3 mg/mL and 6 mg/mL for both spices. Phytochemical analysis revealed higher total phenolic and flavonoid contents in clove, with hydroalcoholic extracts showing greater concentrations than aqueous extracts. HPLC quantified higher eugenol content in clove extracts and higher piperine content in black pepper extracts. The differences in bioactive compound content were statistically significant (p < 0.05). Conclusion: The study confirms that both spices possess significant antimicrobial properties, attributable to their rich phytochemical composition, particularly phenolics and flavonoids. Clove exhibited slightly superior antimicrobial activity compared to black paper. These findings support the potential use of these spices as complementary antimicrobial agents. Further research should investigate their synergistic effects with conventional antibiotics and explore their applications in food preservation and alternative medicine.

Terpenoids from Marine Sources: A Promising Avenue for New Antimicrobial Drugs.

Currently, there is an urgent need for new antibacterial and antifungal agents to combat the growing challenge of antibiotic resistance. As the largest ecosystem on Earth, the marine ecosystem includes a vast array of microorganisms (primarily bacteria and fungi), plants, invertebrates, and vertebrates, making it a rich source of various antimicrobial compounds. Notably, terpenoids, known for their complex structures and diverse bioactivities, are a significant and promising group of compounds in the battle against bacterial and fungal infections. In the past five years, numerous antimicrobial terpenoids have been identified from marine organisms such as bacteria, fungi, algae, corals, sea cucumbers, and sponges. This review article provides a detailed overview of 141 terpenoids with antibacterial and/or antifungal properties derived from marine organisms between 2019 and 2024. Terpenoids, a diverse group of natural organic compounds derived from isoprene units, are systematically categorized based on their carbon skeleton structures. Comprehensive information is provided about their names, structures, biological sources, and the extent of their antibacterial and/or antifungal effectiveness. This review aims to facilitate the rapid identification and development of prospective antimicrobials in the pharmaceutical sector.

Detection of multidrug-resistance in Mycobacterium tuberculosis by phenotype- and molecular-based assays.

BACKGROUND: The whole-genome sequencing (WGS) is becoming an increasingly effective tool for rapid and accurate detection of drug resistance in Mycobacterium tuberculosis complex (MTBC). This approach, however, has still been poorly evaluated on strains from Central and Eastern European countries. The purpose of this study was to assess the performance of WGS against conventional drug susceptibility testing (DST) for the detection of multi-drug resistant (MDR) phenotypes among MTBC clinical strains from Poland and Lithuania. METHODS: The study included 208 MTBC strains (130 MDR; 78 drug susceptible), recovered from as many tuberculosis patients in Lithuania and Poland between 2018 and 2021. Resistance to rifampicin (RIF) and isoniazid (INH) was assessed by Critical Concentration (CC) and Minimum Inhibitory Concentration (MIC) DST as well as molecular-based techniques, including line-probe assay (LPA) and WGS. The analysis of WGS results was performed using bioinformatic pipeline- and software-based tools. RESULTS: The results obtained with the CC DST were more congruent with those by LPA compared to pipeline-based WGS. Software-based tools showed excellent concordance with pipeline-based analysis in prediction of RIF/INH resistance. The RIF-resistant strains demonstrated a relatively homogenous MIC distribution with the mode at the highest tested MIC value. The most frequent RIF-resistance conferring mutation was rpoB S450L. The mode MIC for INH was two-fold higher among double katG and inhA mutants than among single katG mutants. The overall rate of discordant results between all methods was calculated at 5.3%. Three strains had discordant results by both genotypic methods (LPA and pipeline-based WGS), one strain by LPA only, three strains by MIC DST, two strains by both MIC DST and pipeline-based WGS, and the remaining two strains showed discordant results with all three methods, compared to CC DST. CONCLUSIONS: Considering MIC DST results, current CCs of the first-line anti-TB drugs might be inappropriately high and may need to be revised. Both molecular methods demonstrated 100% specificity, while pipeline-based WGS had slightly lower sensitivity for RIF and INH than LPA, compared to CC DST.

Average Nucleotide Identity and Digital DNA-DNA Hybridization Analysis Following PromethION Nanopore-Based Whole Genome Sequencing Allows for Accurate Prokaryotic Typing.

Whole-genome sequencing (WGS) is revolutionizing clinical bacteriology. However, bacterial typing remains investigated by reference techniques with inherent limitations. This stresses the need for alternative methods providing robust and accurate sequence type (ST) classification. This study optimized and evaluated a GridION nanopore sequencing protocol, adapted for the PromethION platform. Forty-eight Escherichia coli clinical isolates with diverse STs were sequenced to assess two alternative typing methods and resistance profiling applications. Multi-locus sequence typing (MLST) was used as the reference typing method. Genomic relatedness was assessed using Average Nucleotide Identity (ANI) and digital DNA-DNA Hybridization (DDH), and cut-offs for discriminative strain resolution were evaluated. WGS-based antibiotic resistance prediction was compared to reference Minimum Inhibitory Concentration (MIC) assays. We found ANI and DDH cut-offs of 99.3% and 94.1%, respectively, which correlated well with MLST classifications and demonstrated potentially higher discriminative resolution than MLST. WGS-based antibiotic resistance prediction showed categorical agreements of ≥ 93% with MIC assays for amoxicillin, ceftazidime, amikacin, tobramycin, and trimethoprim-sulfamethoxazole. Performance was suboptimal (68.8-81.3%) for amoxicillin-clavulanic acid, cefepime, aztreonam, and ciprofloxacin. A minimal sequencing coverage of 12× was required to maintain essential genomic features and typing accuracy. Our protocol allows the integration of PromethION technology in clinical laboratories, with ANI and DDH proving to be accurate and robust alternative typing methods, potentially offering superior resolution. WGS-based antibiotic resistance prediction holds promise for specific antibiotic classes.

Comparative Evaluation of the Antimicrobial Efficacy of Elettaria cardamomum (0.5%) Mouthwash, Camellia sinensis (0.5%) Mouthwash, and 0.12% Chlorhexidine Gluconate Mouthwash against Streptococcus mutans: An In Vitro Study.

Aim: The in vitro study aimed to evaluate and compare the antimicrobial efficacy of Elettaria cardamomum (0.5%) mouthwash, Camellia sinensis (0.5%) mouthwash, and 0.12% chlorhexidine gluconate mouthwash against Streptococcus mutans. Materials and methods: A total of 60 samples of the five mouthwash preparations were prepared to check for their antimicrobial efficacy. The zone of inhibition (ZOI) against S. mutans was measured as a diameter in mm, and the minimum inhibitory concentration (MIC) of mouthwash preparations was measured as µg/mL. All the groups were compared statistically using the Mann-Whitney U test and the Kruskal-Wallis test. Results: The highest ZOI was observed in group V chlorhexidine gluconate [mean: 20.8, standard deviation (SD): 0.58], followed by group III C. sinensis (alcohol-free) (mean: 15.5, SD: 0.67), group IV C. sinensis (alcohol-based) (mean: 14.08, SD: 0.66), and group II E. cardamomum (alcohol-based) (mean: 13.2, SD: 0.45). The least ZOI was observed in group I E. cardamomum (alcohol-free) (mean: 10.7, SD: 0.45). This difference was statistically significant (p < 0.01). The MIC was similar in all the groups (p = 0.13). Conclusion: Chlorhexidine gluconate 0.12% mouthwash showed the best antimicrobial action; however, C. sinensis mouthwash showed potential against S. mutans. E. cardamomum mouthwash exhibited limited antimicrobial activity. How to cite this article: Deolikar S, Jawdekar A, Saraf T, et al. Comparative Evaluation of the Antimicrobial Efficacy of Elettaria cardamomum (0.5%) Mouthwash, Camellia sinensis (0.5%) Mouthwash, and 0.12% Chlorhexidine Gluconate Mouthwash against Streptococcus mutans: An In Vitro Study. Int J Clin Pediatr Dent 2024;17(4):461-466.