Minimum Inhibitory Concentration Values Research Articles

Comparative evaluation of MIC values of Trichosporon spp. by MTT assay and CLSI M27-A3 broth microdilution reference methods

Objectives: The objective of this study was to determine and compare the minimum inhibitory concentration (MIC) values of Trichosporon spp. by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl- 2H-tetrazoliumbromide) assay, and Clinical and Laboratory Standards Institute M27-3rd edition (CLSI M27-A3) broth microdilution methods. Materials and Methods: Antifungal susceptibility testing was done by CLSI M27-A3 broth microdilution and MTT assay for all the 72 Trichosporon isolates after genus specific and Trichosporon asahii specific polymerase chain reaction (PCR). Candida krusei ATCC 6258 was used as the reference strain. Statistical Analysis: All statistical data were analyzed using the Statistical Package for the Social Sciences, version 17 for Microsoft Windows. The percentage of agreement was calculated using the Type C intraclass correlation coefficient. Results: The MICs by MTT assay strongly correlated with those obtained by CLSI M27-A3 method, by being either the same or within 1 dilution of MIC by CLSI method. Furthermore, the ranges of MICs obtained by MTT and CLSI method were all identical in our study. The overall agreement between the two methods for the Trichosporon isolates was good, that is, 90.8% in our study. Conclusions: MTT assay can be an alternative method that assists reading of MICs visually with a colored end point, making it easier compared to CLSI M27-A3 method. MTT assay can also be standardized for other yeasts and molds so that antifungal susceptibility tests can be done for different fungi.

Antibacterial Activity of Ethanol Extract of Aceh Patchouli Leaves (Pogostemon cablin Benth.) against Enterococcus faecalis: A Potential Alternative for Root Canal Infections

The persistence of bacterial infections and antibiotic resistance in secondary root canal infections necessitate effective antimicrobial approaches utilizing natural compounds. This study aimed to evaluate the extraction efficiency, phytochemical composition, and antibacterial activity of the ethanol extract of Aceh patchouli leaves (Pogostemon cablin Benth.) against Enterococcus faecalis. The maceration method was employed for extraction and phytochemical analysis was conducted using gas chromatography-mass spectrometry (GC-MS). Antibacterial activity was assessed using the Kirby-Bauer method, and the minimum inhibitory concentration (MIC) was determined using UV-vis spectrophotometry. The results indicated a total extract yield of 17.25 % and significant antibacterial activity against E. faecalis. The active compounds in the extract were primarily sesquiterpenes and fatty acids, with patchouli alcohol as the most prominent. The MIC value was 12.5 %, suggesting the potential of Aceh patchouli extract as an alternative or adjunctive treatment for root canal infections. This study has the potential to make Aceh's indigenous Pogostemon cablin Benth. plant as an alternative or adjunctive treatment for root canal infections leading to safe and effective treatments that minimize the risk of antibiotic resistance. HIGHLIGHTS The ethanol extract of Aceh patchouli leaves (Pogostemon cablin Benth.) was obtained using the maceration method, yielding a total extract value of 17.25 %. The active compounds of the extract were primarily sesquiterpenes and fatty acids, with patchouli alcohol being the most prominent. The ethanol extract exhibited significant antibacterial activity against Enterococcus faecalis, a common bacterium responsible for secondary root canal infections. The MIC of the extract was 12.5 %, indicating its potential effectiveness as an antimicrobial agent. This study suggests that the Aceh patchouli leaves could serve as an alternative or adjunctive treatment for root canal infections, addressing the challenges of bacterial persistence and antibiotic resistance. GRAPHICAL ABSTRACT

Isolation and characterization of Streptococcus agalactiae inducing mass mortalities in cultured Nile tilapia (Oreochromis niloticus) with trials for disease control using zinc oxide nanoparticles and ethanolic leaf extracts of some medicinal plants

BackgroundStreptococcus agalactiae (Group B streptococcus, GBS) induces a serious infection that can harm not only aquatic life but also humans and other animals. In a fish farm in southern Egypt, Nile tilapia (Oreochromis niloticus) has developed an epidemic with clinical symptoms resembling piscine streptococcosis.ResultsInitial microscopic inspection of the affected fish brain and kidney indicated the presence of Gram-positive cocci. S. agalactiae was effectively isolated and identified using nucleotide homology of the 16S rRNA and species-specific PCR. The partial 16S rRNA sequence was deposited in the GenBank database at the NCBI and given the accession number MW599202. Genotyping using RAPD analysis indicated that the isolates in the present study belonged to the same genotypes and had the same origin. The challenge test, via immersion (9.2 × 107, 9.2 × 106, and 9.2 × 105 CFU/ml for 1 h) or intraperitoneal injection (4.6 × 107, 4.6 × 106, and 4.6 × 105 CFU/fish), elicited clinical symptoms resembling those of naturally infected fish with a mortality rate as high as 80%. The ability to create a biofilm as one of the pathogen virulence factors was verified. Zinc oxide nanoparticles and the ethanolic leaf extracts of nine medicinal plants demonstrated considerable antibacterial activities against the tested S. agalactiae strain with low minimum bactericidal concentrations (MBC) and minimum inhibitory concentrations (MIC). The ethanolic leaf extracts from Lantana camara and Aberia caffra showed potent antibacterial activity with MBC values of 0.24 and 0.485 mg/ml, and MIC values of 0.12 & 0.24 mg/ml, respectively.ConclusionThis study isolated S. agalactiae from O. niloticus mortalities in a fish farm in Assiut, Egypt. The pathogen persists in fish environments and can escape through biofilm formation, suggesting it cannot be easily eliminated. However, promising findings were obtained with in vitro control employing zinc oxide nanoparticles and medicinal plant extracts. Nevertheless further in vivo research is needed.

Anti-Trichophyton rubrum potential, association with fluconazole, and mechanism of action of (R)-(+)-citronellal

Trichophyton rubrum is a keratolytic and keratinophilic fungus responsible for causing dermatophytoses. These pathogens have been associated with resistance phenomena, which encourages the need to search for new substances with anti-Trichophyton rubrum activity. The present study aimed to investigate the antifungal activity of (R)-(+)- Citronellal (RCIT) against clinical isolates of T. rubrum. The antifungal potential of RCIT was evaluated from the Minimum Inhibitory Concentration (MIC), Minimum Fungicide Concentration (MFC), Association study, and assays with cholesterol, ergosterol and sorbitol. RCIT MIC values ranged from 4 to 512 µg/mL, while their MFC ranged from 4 to 512 µg/mL. When associating RCIT with the drug fluconazole, pharmacological indifference and antagonism were evidenced. It was shown that the mechanism of action is related to fungal ergosterol and showed interactions with exogenous cholesterol. The results obtained in this research demonstrate that RCIT has the potential to become a product for the treatment of dermatophytosis.

Synthesis and evaluation of antibacterial and antibiofilm agents based on phenylamino-substituted 1,4-benzoquinones.

Aim: This work describes the synthesis and antimicrobial evaluation of 6-aminated 1,4-benzoquinones (6-AQs) against seven resistant pathogens.Materials & methods: The 6-AQs, synthesized via a Michael addition reaction between bromoquinone and p-substituted anilines, were assessed for their antimicrobial activity through both in vitro and in silico analyses.Results: Bromoquinone and 6-AQs with electron-withdrawing groups demonstrated activity against Pseudomonas aeruginosa, with minimum inhibitory concentrations ranging from 16 to 128μg/ml, comparable to standard antimicrobials. Two derivatives exhibited minimum inhibitory concentrations values against methicillin-resistant Staphylococcus aureus ranging from 64 to 128μg/ml. These compounds demonstrated both bacteriostatic and bactericidal effects, and antibiofilm features.Conclusion: The 6-AQs 19g and 19f showed a promising antimicrobial profile, indicating their potential as new therapeutic options.

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.

Biosynthesis of CuO nanoparticle using leaf extracts of Ocimum lamiifolium Hochst. ex Benth and Withana somnifera (L) Dunal for antibacterial activity

Nanotechnology is becoming a promise for scientific advancement nowadays in areas like medicine, consumer products, energy, materials, and manufacturing. Copper oxide nanoparticles (CuO NPs) were synthesized using Ocimum lamiifolium Hochst. ex Benth and Withana somnifera (L) Dunal leaf extract via green synthetic pathway. The leaf of O. lamiifolium and W. somnifera were known to have strong antibiotic and antioxidant properties arising due to the presence of various secondary metabolites, including, flavonoids, alkaloids, saponins, tannins, cardiac glycosides, and phenolic compounds which serve as reducing, stabilizing, and capping agents for the CuO-Nanoparticles (NPs) synthesized. The biosynthesized CuO NPs were characterized based on Fourier transform infrared spectroscopy, X-ray diffraction spectroscopy, and scanning electron microscopy. O. lamiifolium and W. somnifera leaf extract mediated synthesis could produce CuO NPs with average crystallite size of 15 nm and 19 nm, respectively. The biosynthesized CuO-NPs were further examined for antibacterial activity with Gram-positive (S. aureus) and Gram-negative bacteria (E. coli and P. aeruginosa). The GZDK-CuO NPs synthesized using W. somnifera leaf extract inhibited the growth of E. coli. and P. aeruginosa largely in comparison to S. aureus. Whereas the DMAZ-CuO NPs synthesized with the help of O. lamiifolium leaf extract showed higher bacterial inhibition on E. coli compared to S. aureus and P. aeruginosa. The minimum inhibitory concentration (MIC) values of both types of NPs are also assessed on all three pathogens. The newly biosynthesized nanoparticles, thus, were found to be optional materials for inhibiting the growth of drug- resistant bacterial pathogens.

Nisin-loaded chitosan/sodium alginate microspheres enhance the antimicrobial efficacy of nisin against Staphylococcus aureus.

To develop and evaluate nisin-loaded chitosan/sodium alginate (CS/SA) microspheres as an improved antimicrobial delivery system targeting Staphylococcus aureus strains. The microspheres were prepared using a modified water-in-oil emulsion cross-linking method, resulting in spherical particles sized 1-8µm with a surface charge of -7.92±5.09mV, confirmed by scanning electron microscopy (SEM) and Zetasizer analysis. Encapsulation efficiency (EE) and loading capacity (LC) of nisin were 87.60±0.43% and 1.99±0.01%, respectively. In vitro release studies over 48 hours indicated a controlled release pattern of nisin, described by the Korsmeyer-Peppas model, with higher release rates at 37°C and alkaline pH. Antimicrobial assays showed an enhanced efficacy of nisin-loaded CS/SA microspheres compared to free nisin, with minimum inhibitory concentration (MIC) values reduced by 50%. Confocal laser scanning microscopy (CLSM), SEM and transmission electron microscopy (TEM) showed significant bacterial membrane damage and cellular disruption induced by the microspheres. This study highlights the potential of nisin-loaded CS/SA microspheres as an innovative antimicrobial delivery system with improved stability and antimicrobial efficacy against S. aureus, addressing limitations associated with nisin alone.

HPLC analysis and antimicrobial activity of Vitex altissima leaves extracts

The exploration of ethnomedicinal plants for phytochemical and pharmacological properties is crucial for developing novel therapeutics for chronic diseases. Vitex altissima, known as the Peacock chaste tree, is a significant member of the Verbenaceae family, is widely utilized in traditional medicine. This study investigates the phytochemical composition and antibacterial properties of V. altissima, a plant recognized for its medicinal use by various indigenous communities, including the Malayali tribes of Servarayan hills. The study involved extracting phytochemicals from the leaves of V. altissima using hexane, ethyl acetate and methanol through soxhlet extraction. The extracts were analyzed for secondary metabolites, including carbohydrates, proteins, amino acids, steroids, glycosides, alkaloids, tannins, phenolics, flavonoids, terpenoids, saponins, anthraquinones, oils and resins, diterpenes, phlobatannins and coumarins. Quantitative assessments of phenolic compounds, flavonoids and alkaloids were performed. Column chromatography was employed to fractionate the methanolic extract, resulting in 12 distinct fractions. These fractions were screened for antibacterial activity against E. coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus mutans and Staphylococcus aureus using the agar well diffusion method. The minimal inhibitory concentration (MIC) was determined through a 2-fold serial dilution technique. High-performance liquid chromatography (HPLC) was utilized to identify the polyphenolic compounds responsible for antibacterial activity. Phytochemical screening revealed a diverse array of bioactive compounds in the extracts, with methanol extracts showing the highest total phenolic content (218 ± 7.21 mg GAE/g of extract) and significant antibacterial activity. The ethyl acetate extract exhibited the highest total flavonoid content (40.67 ± 6.65 mg QE/g of extract). Among the fractions, the column fraction from the chloroform-methanol gradient (VACF) demonstrated superior antimicrobial activity, particularly against S. mutans and P. aureginosa. The MIC values for VACF were 427 µg/mL for P. aeruginosa and 400 µg/mL for S. aureus, indicating potent antibacterial properties. HPLC analysis identified key polyphenolic compounds, including p-coumaric acid, ferulic acid and elagic acid, as the primary contributors to the antibacterial activity. The presence of these compounds aligns with the observed antimicrobial efficacy and highlights the potential of V. altissima extracts as a source of natural antibacterial agents.

Antifungal efficacy of photodynamic therapy on Cryptococcus and Candida species is enhanced by Streptomyces spp. extracts in vitro.

The research on actinobacteria isolated from traditional medicinal plants is limited. Here, four new Streptomyces isolates (Ha1, Pp1, UzK and UzM) were obtained from the rhizospheres of Helianthus annuus, Pongamia pinnata and Ziziphus mauritiana, frequently utilized in Indian traditional medicine. The Streptomyces isolates aqueous extracts were studied alone against the growth of the Cryptococcus neoformans H99 reference strain, the fluconazole-tolerant T1-5796 and 89-610 strains, three histone deacetylase (HDAC) genes mutant strains, C. gattii NIH198, Candida albicans, C. glabrata, C. parapsilosis and C. tropicalis to determine minimum inhibitory concentration (MIC). Next, the extracts were employed in combination with aluminium-phthalocyanine chloride nanoemulsion-mediated photodynamic therapy to evaluate a possible interaction. We demonstrated that the C. neoformans T1-5796 fluconazole-tolerant strain was more severely inhibited by the Pp1 isolate extract (MIC: 6mg mL-1) than H99, which was not inhibited. Growth inhibition of the HDAC null mutants was more prominent for the extract of the UzM isolate, showing inhibition at 2mg mL-1. The UzM extract was also the most effective in hindering the Candida species proliferation, with MIC values ranging from 10 to 40mg mL-1. The four Streptomyces extracts, especially UzK and UzM, significantly enhanced the antifungal effect of the photodynamic therapy. Our results indicate these Streptomyces isolates as sources of novel metabolites which could potentiate the effect of photodynamic therapy in controlling yeasts superficial infections.

Monomelittoside and Dichotomoside D from bark extract of Alstonia scholaris (L.) R. Br. inhibits dental caries pathogen Streptococcus mutans

Dental caries advances to be a significant global health concern, primarily attributed to the virulent actions of Streptococcus mutans. The antibacterial property of the bark extract of Alstonia scholaris was assessed using standard disc diffusion assays, which revealed significant inhibition of S. mutans. Subsequently, we determined the Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) revealed potent antibacterial activity of bark extract against S. mutans, at MIC and MBC values 62.5 and 125 µg/mL, respectively and antibiofilm activity at 125 µg/mL. Furthermore, cytotoxicity assays showed no adverse effects on mammalian cells. Moreover, we employed liquid chromatography–mass spectrometry to identify bioactive compounds in the bark extract and revealed the presence of major active compounds as Monomelittoside and Dichotomoside D effective in targeting the S. mutans. These findings showed that, the promising antibacterial efficacy of Monomelittoside and Dichotomoside D, highlighting their potential as natural therapeutics for combating dental caries.

Effect of solvent polarity on yield extract, antioxidant and antibacterial activities of phytochemicals from Andrographis paniculata leaves

ABSTRACT In this study, the impact of solvent polarity on the extraction yield, antibacterial activity and antioxidant characteristics of phytochemical compounds derived from Andrographis paniculata leaves was investigated. The simplicia of A. paniculata was macerated in different organic solvents with raising polarity (n-Hexane, ethyl acetate, dichloromethane, ethanol, methanol, methanol–water) at room temperature for 24 h. Preliminary screening revealed that the yield extract, phytochemical composition, antibacterial activity and antioxidant properties were influenced by the polarity of the solvent used for extraction. The optimum yield extract was obtained using a methanol–water solvent (50:50, v/v) of 15.46%. The most active antibacterial activity against biofilm-forming bacteria (Pseudomonas aeruginosa) was obtained using ethyl acetate and dichloromethane with a minimum inhibition concentration (MIC) value of 312.5 µg mL−1. Meanwhile, the antioxidant properties of A. paniculata leaf extract with all solvents shows a very weak level. The antibacterial activity and antioxidant properties of A. paniculata leaf extracts are related to the phytochemical content in the extract. This result is evident in the extract with ethyl acetate solvents with the highest total phenolic and flavonoid values. However, GC-MS analysis of A. paniculata extract revealed 20 components, with Supraene having the greatest compounds.

Assessment of the activity and mechanisms of resistance to cefiderocol and combinations of β-lactams and the novel β-lactamase inhibitors avibactam, taniborbactam, zidebactam, nacubactam, xeruborbactam, and ANT3310 in emerging double-carbapenemase-producing Enterobacterales.

We aimed to investigate the activity of and mechanisms of resistance to cefiderocol and innovative β-lactam/β-lactamase inhibitor combinations in a nationwide collection of double-carbapenemase-producing Enterobacterales. In all, 57 clinical isolates co-producing two carbapenemases collected from Spanish hospitals during the period 2017-2022 were analyzed. Minimum inhibitory concentration (MIC) values for ceftazidime, ceftazidime/avibactam, aztreonam, aztreonam/avibactam, aztreonam/nacubactam, cefiderocol, cefepime, cefepime/taniborbactam, cefepime/zidebactam, cefepime/nacubactam, imipenem, imipenem/relebactam, meropenem, meropenem/vaborbactam, meropenem/xeruborbactam, and meropenem/ANT3310 were determined by reference broth microdilution. Genetic drivers of resistance were analyzed by whole-genome sequencing (WGS). The collection covered nine carbapenemase associations: VIM + OXA-48 (21/57), NDM + OXA-48 (11/57), KPC + VIM (10/57), KPC + OXA-48 (6/57), IMP + OXA-48 (3/57), NDM + KPC (2/57), NDM + VIM (2/57), NDM + GES (1/57), and KPC + IMP (1/57). Ceftazidime/avibactam, imipenem/relebactam, and meropenem/vaborbactam were the least active options. Aztreonam/avibactam and aztreonam/nacubactam were active against the whole collection and yielded MIC50/MIC90 values of ≤0.25/0.5 mg/L and 1/2 mg/L, respectively. Cefepime/zidebactam (56/57 susceptible), meropenem/xeruborbactam (56/57 susceptible), cefepime/nacubactam (55/57 susceptible), and cefiderocol (53/57 susceptible) were also highly active, with MIC50/MIC90 values ranging from ≤0.25-2 mg/L to 2-4 mg/L, respectively. Meropenem/ANT3310 (MIC50/MIC90 = 0.5/≥64 mg/L; 47/57 susceptible) and cefepime/taniborbactam (MIC50/MIC90 = 0.5/16 mg/L; 44/57 susceptible) also retained high levels of activity, although they were affected by NDM-type enzymes in combination with porin deficiency. Our findings highlight that cefiderocol and combinations of β-lactams and the novel β-lactamase inhibitors avibactam, nacubactam, taniborbactam, zidebactam, xeruborbactam, and ANT3310 show promising activity against double-carbapenemase-producing Enterobacterales.

Multi-Locus Sequence Typing-Based Genetic Analysis, Antifungal Resistance, and Clinical Prognosis of Cryptococcus neoformans Infections in HIV-Infected Patients in Northern China.

This study aimed to investigate the molecular epidemiological characteristics and drug sensitivity of Cryptococcus from HIV-infected patients and their relationship with patients' prognosis. Seventy-six strains were collected and identified to the species level by matrix-assisted laser desorption ionization-time of flight mass spectrometry, confirmed by internal transcribed spacer sequencing. Multi-locus sequence typing was used for the typing of Cryptococcus, and its antifungal susceptibility was tested using FUNGUS 3. The clinical outcomes of the patients were reviewed at 3-, 6-, 9-, and 12-month follow-ups. All strains were Cryptococcus neoformans var. grubii classified into seven sequence types (STs) dominated by ST5, ST31, and a new ST702 strain. The 6- and 9-month survival rates were highest for patients infected with ST31, ST32, and ST174. The antifungal resistant rates were 13.2%, 2.6%, and 1.4% for fluconazole, amphotericin B, and 5-fluorocytosine. Except itraconazole, the minimum inhibitory concentration (MIC) values and wild type (WT)/non-wild type (NWT) of Cryptococcus for antifungal drugs were not related to the clinical prognosis of HIV-infected patients with cryptococcal infection. ST5 was the main ST type, and the new ST702 type was found in a patient who died in a short period of time. Cryptococcus neoformans var. grubii had a relatively high antifungal drug resistance rate to fluconazole. The WT strain accounted for the highest proportions for 5-fluorocytosine, amphotericin B, fluconazole, voriconazole, and itraconazole. The MIC values of Cryptococcus for first-line antifungal drugs showed no relationship with clinical prognosis, implying that MIC values cannot be used to predict the clinical outcome of these patients.

Recent increase in Candida auris frequency in the SENTRY surveillance program: antifungal activity and genotypic characterization.

We observed an increase in the frequency of Candida auris among invasive candidiasis isolates in the 2022 SENTRY Antifungal Surveillance Program compared to prior years: ≤0.1% before 2018, 0.4%-0.6% from 2018 to 2021, and 1.6% in 2022. C. auris isolates were collected in seven countries, but 28 (35.9%) isolates were recovered in the USA (five states; more common in New York, Texas, and New Jersey) and 26 (33.3%) in Panama. Greece and Turkey had 12 and 9 isolates, respectively. Overall, 82.1% of the isolates were resistant to fluconazole; 17.9% were resistant to amphotericin B; and 1.3% were resistant to caspofungin, anidulafungin, or micafungin (Centers for Disease Control and Prevention tentative resistance breakpoints). Rezafungin inhibited 96.2% of the isolates (Clinical and Laboratory Standards Institute susceptibility breakpoint). Pandrug resistance was not observed, but 17.9% of the isolates were resistant to fluconazole and amphotericin B. South Asian (Clade I) isolates were most common (n = 40, 51.3%); of these, 97.5% were resistant to fluconazole and 30.0% were resistant to amphotericin B. Thirty (38.5%) isolates belonged to the South American region (Clade IV), and 56.7% of those were resistant to fluconazole and 6.7% to amphotericin B. Seven isolates belonged to the South African Clade III and one to East Asian Clade II. Erg11 (Y132F, K143R, and F126L) and MRR1 (N647T) alterations were detected. One isolate that was resistant to all echinocandins carried an FKS R1354G alteration. Two isolates displayed elevated rezafungin minimum inhibitory concentration (MIC) values but low MIC values against other echinocandins and no FKS alterations. As C. auris is spreading globally, monitoring this species is prudent.

The chemical composition, antioxidative and antimicrobial potentials of Toona sinensis essential oils from different regions of China

Toona sinensis is one of the traditional Chinese medicinal materials with rich physiological activities. This study aimed to investigate the chemical composition differences of the essential oils from the tender leaves and shoots of T. sinensis, collected from four main producing areas in China, and to verify their anti-listeria and antioxidant properties. Gas chromatography-mass spectrometry (GC-MS) analysis was conducted on the essential oils obtained by hydro-distillation. The results indicated that compounds such as β-elemene (3.5∼8.1%), α-caryophyllene (3.7∼7.2%), and trans-caryophyllene (4.0∼9.7%) were the main common components of T. sinensis essential oils (TSEO) from four producing areas. The results of DPPH and ABTS showed that the essential oil from Dazhou, Sichuan, China had the best antioxidant effect. At a concentration of 1.25 mg/mL, the antioxidant activity of TSEO-DZ from Sichuan was comparable to 0.08 mg/mL of BHT and was only slightly lower than 0.06 mg/mL of VC. The broth dilution method showed that the essential oil from Tai ‘an, Shandong, China, showed better antibacterial effect, with the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MLC) values of 1.25 μL/mL and 2.54 μL/mL, respectively. This study demonstrates that TSEO possesses promising application potential in antioxidant and anti-listeria activities, and it confirms that the growth environment of T. sinensis significantly influences the quality, content, and biological activity of the essential oil.

Innovative Microencapsulation of Polymyxin B for Enhanced Antimicrobial Efficacy via Coated Spray Drying.

This study aims to develop an innovative microencapsulation method for coated Polymyxin B, utilizing various polysaccharides such as hydroxypropyl β-cyclodextrin, alginate, and chitosan, implemented through a three-fluid nozzle (3FN) spray drying process. High-performance liquid chromatography (HPLC) analysis revealed that formulations with a high ratio of sugar cage, hydroxypropyl β-cyclodextrin (HPβCD), and sodium alginate (coded as ALGHCDHPLPM) resulted in a notable 16-fold increase in Polymyxin B recovery compared to chitosan microparticles. Morphological assessments using fluorescence labeling confirmed successful microparticle formation with core/shell structures. Alginate-based formulations exhibited distinct layers, while chitosan formulations showed uniform fluorescence throughout the microparticles. Focused beam reflectance and histograms from fluorescence microscopic measurements provided insights into physical size analysis, indicating consistent sizes of 6.8 ± 1.2 μm. Fourier-transform infrared (FTIR) spectra unveiled hydrogen bonding between Polymyxin B and other components within the microparticle structures. The drug release study showed sodium alginate's sustained release capability, reaching 26 ± 3% compared to 94 ± 3% from the free solution at the 24 h time point. Furthermore, the antimicrobial properties of the prepared microparticles against two Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, were investigated. The influence of various key excipients on the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values was evaluated. Results demonstrated effective bactericidal effects of ALGHCDHPLPM against both E. coli and P. aeruginosa. Additionally, the antibiofilm assay highlighted the potential efficacy of ALGHCDHPLPM against the biofilm viability of E. coli and P. aeruginosa, with concentrations ranging from 3.9 to 500 μg/m. This signifies a significant advancement in antimicrobial drug delivery systems, promising improved precision and efficacy in combating bacterial infections.

SPECIES IDENTIFICATION AND IN VITRO ANTIFUNGAL SUSCEPTIBILITY TESTING OF ASPERGILLUS ISOLATED FROM VARIOUS CLINICAL SAMPLES

Objectives: Aspergillus is ubiquitous mold species present in environment in the form of spores. Infection due to Aspergillus is uncommon in immunocompetent individuals unless they possess any abnormality or have undergone any treatment with corticosteroids. In immunocompromised individuals, infection by this fungus is common among people with prolonged neutropenia, transplants, and human immunodeficiency virus infection. The symptoms are diverse, ranging from allergic reactions to invasive aspergillosis and life-threatening complications. The aim of this study is to isolate and identify Aspergillus species obtained from various clinical specimens and perform antifungal susceptibility testing (AFST). Methods: A total of 200 isolates in which positive direct microscopic findings correlated well with culture growth for Aspergillus were included. Antifungal susceptibility was performed by micro broth dilution technique according to clinical laboratory standard institute M38-A2 guidelines. Results: Aspergillus flavus was found to be predominant species followed by Aspergillus fumigatus. AFST was performed, where all Aspergillus strains exhibited low minimum inhibitory concentration (MIC) and minimum effective concentration (MEC) values for most of antifungal drugs tested except for one strain of A. flavus, which exhibited high MIC for voriconazole and high MEC for caspofungin. Conclusion: Polyenes, azoles, and echinocandin resistance are emerging in many parts of the world. Therefore, continued application of AFST combined with morphological characterization for species identification is critical in detecting the emergence of resistance among various Aspergillus species to reduce mortality, morbidity, and overcome treatment failure.

Synergistic invitro antimicrobial activity of polyherbal combination of Morinda lucida fruit and Pterocarpus santalinoides seed against multi-drug resistant clinical bacterial isolates

Background: Synergistic drug combination has been shown to be a way of bypassing drug resistance and reducing the amounts of antimicrobials consumed. As infections caused by multi-drug resistant organisms (MDROs) continue to pose global threat, it is important to search for new antimicrobial combinations of plant origin that are safe and readily available. The objective of this study is to evaluate the synergistic antimicrobial activity of extracts of Morinda lucida fruit and Pterocarpus santalinoides seed against multi-drug resistant (MDR) bacterial isolates Methodology: Morinda lucida and P. santalinoides fruits were plucked and washed clean, and the fruits of P. santalinoides were deseeded to remove the seeds. They were cut into smaller pieces and dried under shade before being milled into smooth powder and extracted with methanol. The clinical bacterial isolates used were MDR Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. The phytochemical constituents of the extracts were determined using the standard method. The invitro antimicrobial assay of the extracts was done at a concentration of 50 to 400 mg/ml using the agar well diffusion technique. The minimum inhibitory concentration (MIC) and fractional inhibitory concentration (FIC) of each extract were determined using the Checker-board micro-titration method, and the FIC result obtained for each isolate was used to calculate the fractional inhibitory concentration index (FICI) of the combined extracts. The time kill assay was performed to confirm the synergistic and bactericidal activities from the MIC values obtained. Results: The phytochemical analysis revealed that the extracts of both herbal plants contained alkaloids, phenol, tannin, saponin, glycosides, and terpenoids. The antimicrobial assay results showed that the polyherbal combination of the extracts was active against all the MDR bacterial isolates tested with varying zones of inhibition. The mean inhibition zone diameters of individual M. lucida and P. santalinoides ranged from 18.0±0.9mm to 26.0±1.4mm and 17.0±0.1mm to 24.0±0.2mm respectively, while the MIC ranged from 6.25mg/ml to 12.5mg/ml for M. lucida and 12.5 mg/ml for P. santalinoides. The mean inhibition zone diameter of the polyherbal combination of the two plant extracts ranged from 25.00±00mm to 34.0±0.4mm, which compared favorably with that of levofloxacin control (28.0±0.0mm to 32.00±0.0mm), while their MIC ranged from 0.39mg/ml to 1.56mg/ml. The FIC of M. lucida extract ranged from 0.06mg/ml to 0.25mg/ml while that of P. santalinoides ranged from 0.03mg/ml to 0.13mg/ml. The FICI of combined extracts ranged from 0.09mg/ml to 0.38 mg/ml for all the MDR isolates, which is less than 0.5mg/ml, indicating synergism against all the isolates. The time of kill assay confirmed the synergistic and bactericidal activities with a 3log10 CFU/ml decrease in the number of viable cells within 6 hours of incubation.Conclusion: Our findings showed synergistic antibacterial actions of extracts of M. lucida fruit and P. santalinoides seed against MDR clinical bacterial isolates, comparable to levofloxacin. Combination of these herbal plants may serve as alternative sources of antimicrobial agents for the treatment of infections caused by MDR bacterial pathogens.

Antibacterial and anticancer potential of bioactive compounds and secondary metabolites of endophytic fungi isolated from Anethum graveolens.

Fungal endophytes are known to produce bioactive chemicals and secondary metabolites that are often identical to those produced by their host plants. The main objective of the current study was to isolate and identify endophytic fungi associated with the medicinal plant Anethum graveolens, and to investigate their potential antibacterial and anticancer properties. The ethyl acetate extracts from the isolated endophytic fungi, as well as the host plant A. graveolens, were subjected to bioactivity assays to evaluate their antibacterial and anticancer potential against multi-drug resistant bacterial strains and the human hepatocellular carcinoma cell line HepG2. The endophytic fungi isolated and identified from the A. graveolens samples included Diaporthe, Auxarthron, Arthrinium, Aspergillus, Microsporum, Dothiorella, Trichophyton, Lophiostoma, Penicillium, and Trichoderma species. The minimum inhibitory concentration (MIC) assay revealed that the A. graveolens extract exhibited the strongest antibacterial activity, with an MIC value of 4 μg/ml, followed by the Trichoderma sp. (5 μg/ml) and Penicillium sp. (6 μg/ml) extracts. Additionally, the crude extracts of Trichoderma sp., Penicillium sp., and Fusarium sp. demonstrated high anticancer activity against HepG2 cells, with inhibition rates ranging from 89 to 92% at a concentration of 50 μg/ml. Interestingly, the A. graveolens extract showed the most potent anticancer activity, with a 95% inhibition rate against HepG2 cells at the same concentration. These findings highlight the significant potential of endophytic fungi associated with A. graveolens, as a source of bioactive compounds with promising antibacterial and anticancer properties. The results reinforce the hypothesis that medicinal plants and their endophytic fungi can serve as an attractive alternative for the development of novel therapeutic agents, potentially offering a more sustainable and less harmful approach to disease management compared to traditional chemical-based methods.