Determination Of Minimum Inhibitory Concentration Research Articles

Investigating antimicrobial resistance genes in probiotic products for companion animals.

One of the greatest challenges of our time is antimicrobial resistance, which could become the leading cause of death globally within a few decades. In the context of One Health, it is in the common interest to mitigate the global spread of antimicrobial resistance by seeking alternative solutions, alongside appropriate drug selection and responsible use. Probiotics offer a potential avenue to reduce antibiotic usage; however, there is a scarcity of research that examines commercial products in terms of carrying antimicrobial resistance genes (ARGs) involved in resistance development through microbial vectors. Our study investigated 10 commercially available probiotic products for cats and dogs. Initially, we conducted phenotypic testing through determination of minimum inhibitory concentration (MIC) for antibiotics important in animal and public health. Subsequently, we performed next-generation sequencing (NGS) of the products to elucidate the genetic background behind the decrease in phenotypic sensitivity. In total, 19 types of ARGs were identified, with 57.9% being found on plasmids, and in two cases, carriage as mobile genetic elements were found. One of the genes identified was the APH(3')-Ia gene, capable of inactivating aminoglycoside antibiotics through phosphotransferase enzyme production regulation, while the other was the tetS gene, capable of conferring reduced sensitivity to tetracycline antibiotics through target protection. Our findings underscore the importance of approaching antimicrobial resistance investigations from a broader perspective. We suggest that further studies in this area are justified and raise questions regarding the need to extend legally required studies on probiotic products from their use in economic livestock to their use in companion animals.

Antimicrobial Activity of Commercial Essential Oils Available in the Market of Kathmandu

Given the increasing global apprehension surrounding antibiotic resistance, there has been growing interest in exploring natural compounds, specifically essential oils, as potential substitutes for combating microbial pathogens. This research work aimed to evaluate the antimicrobial activity exhibited by commercial essential oils and by diluting them with the carrier oils at equal concentration. The screening of antimicrobial activity was conducted using the agar well diffusion method with eight essential oils. Using the broth dilution method, the antibacterial activity of eight commonly used essential oils: lavender, tea tree, eucalyptus, peppermint, thyme, citrus, rosemary, and cinnamon were assessed. Both Gram-positive and Gram-negative species were selected for antimicrobial activity. The minimum inhibitory concentration (MIC) was determined by the broth dilution method. Following MIC determination, a sub-culture was performed on nutrient agar plates to ascertain the minimum bactericidal concentration (MBC). All the antibiotics used in the study demonstrated sensitivity when subjected to the antibiotic susceptibility test. Thyme, cinnamon and peppermint strongly inhibited the growth of S. epidermidis 46 mm, 30mm, and 34 mm respectively. Findings revealed distinct variations in the MIC values among the different essential oils and bacteria tested. Thyme and tea tree oils exhibited the broadest antibacterial spectrum, inhibiting both Gram-positive and Gram-negative bacteria at relatively low concentrations. Thyme (15.625 μl/ml) and tea tree (31.25 μl/ml) oil demonstrated promising activity against Gram-positive pathogen, while tea tree oil (15.625 μl/ml) displayed better efficacy against Gram-negative bacteria. A significant outcome was observed when the essential oils were diluted with carrier oils, indicating promising results in terms of antibacterial activity. These findings highlight the potential of essential oils, particularly thyme and tea tree oils, as effective natural antimicrobial agents.

Characterization of Chlorhexidine Resistance Among Clinical Isolates of Coagulase-Negative Staphylococcus Species in a Tertiary Care Center, Chennai.

Background Coagulase-negativeStaphylococci(CoNS) are potential pathogens and are often associated with healthcare-associated infections (HAIs). Chlorhexidine (CHX) is the most widely used antiseptic to reduce colonization and infection by allStaphylococci, including CoNS. Resistance to CHXamong CoNS has been observed over the past few years, consequent to its widespread use. Phenotypic tolerance or reduced susceptibility to CHX is conferred by plasmid-mediatedqacgroup of genes, mainlyqacA/Bandsmr, which cause activation of efflux pumps over the bacterial cell wall. This study aims to characterize the phenotypic and genotypic resistance exhibited by CoNS species against CHX. Methods After ethical approval, 148 consecutive, non-repetitive isolates of clinically significant CoNS species of hospitalized patients, isolated from blood samples and exudative specimens, were included in the study. Speciation was performed by conventional biochemical identification and automated methods. Antimicrobial susceptibility testing was performed by disc diffusion technique and for vancomycin by minimum inhibitory concentration (MIC) determination, as per Clinical Laboratory Standards Institute (CLSI) M-100 2023 guidelines. Methicillin resistance was detected using a cefoxitin disc. MIC for CHX was performed by agar dilution method; reduced susceptibility was considered when MIC to CHX ≥4 µg/mL. The simplex polymerase chain reaction (PCR) was carried out with suitable controls to detectqacA/Bandsmr. Statistical analysis was conducted to determine the association ofqacA/Bandsmrgenes with MIC of CHXin the study isolates. Results Fifteen different species of CoNS were obtained from clinical samples. A high percentage of resistance was observed against various classes of antibiotics. Methicillin resistance was observed in 69.6% (103/148) of isolates. Of 148 CoNS, 52.7% (78/148) of isolates exhibited reduced susceptibility to CHX with an MIC ≥4 µg/mL. These isolates exhibited a higher percentage of methicillin resistance (75.6%, 59/78). By PCR, 34.5% (51/148) of isolates carried either or both genes. GeneqacA/Bwas solely detected in 27.02% (40/148) of isolates, of which 14 were CHX-tolerant and the remaining 26 were CHX-susceptible. Genesmrwas solely detected in 4.1% (6/148) of isolates comprising three isolates each in CHX-tolerant and susceptible categories.There were 3.4% (5/148) of isolates that harbored bothgenes, of which only one isolate was CHX-susceptible, while the other four were CHX-tolerant. A proportion of isolates that were phenotypically tolerant to CHX did not carry either or both genes. A significant statistical association was found between reduced susceptibility to CHX and the presence of antiseptic resistance genesin the study isolates (p-value=0.033942). Conclusion To our knowledge, this is the first study from South India to investigate CHX resistance among CoNS using phenotypic and genotypic methods. The rise of antiseptic resistance among CoNS is an emerging threat to current infection control practices. The presence ofqacA/Bandsmrgenes, especially in CHX susceptible isolates, is concerning since these resistance genes are located on transferable plasmids, and the isolates can develop resistance eventually upon exposure to CHX.

A rare case report of tissue infection caused by Pantoea piersonii (basionym Kalamiella piersonii).

In 2019, Pantoea piersonii was initially isolated from the interior surfaces of the International Space Station. This microorganism is a species within the genus Pantoea in the family Erwiniaceae, belonging to the order Enterobacterales. Recent literature has documented four cases of its isolation. Despite initial predictions suggesting the non-pathogenicity of P. piersonii strains, evidence from observed cases indicates potential pathogenicity. According to documented evidence in the literature, this microorganism is capable of causing severe and life-threatening conditions, including sepsis. Traditional tests, as well as automated systems, may fail to provide complete differentiation due to these similarities. While MALDI-TOF MS is a valuable tool for identification in clinical diagnostic microbiology, sequencing may be necessary for precise identification. To determine the antibiotic susceptibility profile, various methods can be utilized, including minimum inhibitory concentration determination, disk diffusion testing (Kirby-Bauer test), genotypic resistance assays (PCR and sequencing), and automated systems. The literature reports a limited number of cases associating P. piersonii with human infection. This study contributes to this body of knowledge by reporting a novel case in which P. piersonii was isolated from a tissue sample for the first time. In this case report, the patient achieved recovery following the administration of appropriate antibiotic treatment based on the diagnosis. It underscores the need for precise identification and understanding of its pathogenicity.

Characterization of Bacterial Isolates from Freshwater Environments in Livingstone Island, Antarctica, Using Cultivation and Molecular Methods

The present research work summarizes the investigation of the microbial characteristics of bacterial isolates obtained from freshwater samples collected at various points throughout the Bulgarian Antarctic base “St. Kliment Ohridski” on Livingstone Island. The ability of microorganisms in psychrophilic envi¬ronments to survive and reproduce at low temperatures, lack of nutrients, and high UV radiation, suggests that they have developed specific mechanisms to overcome these extreme conditions. To understand the range of influence of these environmental factors standard microbiological techniques were used. The re¬search provides data on the influence of UV light, temperature range, determination of minimum inhibitory concentration (MIC) to different heavy metals, and 16S rDNA taxonomic identification of the isolates. Ant¬arctic microorganisms are of great interest in food, textile, and brewing industries, production of detergents, etc. The microbial composition in such hostile habitats still little affected by the anthropogenic factor is es¬sential for understanding the potential of psychrophilic microorganisms in different practical applications.

Exploring the Potential of New 7‐Chloroquinoline‐benzylamine Hybrids as Antimicrobials: Synthesis, Biological Activity and MD Simulation Studies

AbstractNovel drug like chloroquinoline‐benzylamine hybrids were synthesized and subjected to comprehensive antibacterial evaluation in the present investigation. The pharmacological evaluation comprises determination of minimum inhibitory concentration (MIC), disk diffusion assay, hemolysis and combination assays against both Gram‐negative (Pseudomonas aeruginosa) and Gram‐positive (Bacillus subtilis, Enterococcus faecalis, and Staphylococcus aureus) bacterial strains. Among all, 7‐chloroquinoline‐benzylamine hybrid bearing p‐bromophenyl substituent (SA11) demonstrated significant antibacterial efficacy (MIC=128μg/mL) against the tested panel of Gram‐positive strains with no sign of toxicity towards human red blood cells (hRBCs). Furthermore, another hybrid with o‐hydroxyphenyl substitution (SA12) exhibited notable activity against the tested isolates with MIC values ranging from 64 to 256μg/mL. Molecular docking studies suggested compound SA11 binds within the active site of the biofilm causing protein (PDB: 7C7U) further supported by the molecular dynamics (MD) simulation studies at 100 ns. Compound SA11 exhibited significant efficacy in inhibiting S. aureus growth in the disk diffusion assay. Moreover, it displayed a synergistic effect when combined with ciprofloxacin (CIP), implying its potential utility in addressing antibiotic‐resistant bacterial strains through combination therapy. The pkCSM‐Pharmacokinetics assessment indicated favourable ADME profiles for SA11, supporting its potential as a viable drug candidate.

Evaluation of gradient strip diffusion for susceptibility testing of aztreonam-avibactam in metallo-β-lactamase-producing Enterobacterales.

The emergence of metallo-β-lactamase (MBL)-producing Enterobacterales presents unique clinical treatment challenges. Recently developed β-lactam/ β-lactamase inhibitor combination agents, while effective against other carbapenemase-producing organisms, are notably ineffective against MBL producers. While MBLs do not hydrolyze monobactams (aztreonam), many MBL-producing organisms are resistant to aztreonam through alternate mechanisms, leaving cefiderocol as the sole monotherapy treatment option recommended for MBL producers. Recent guidelines for the treatment of MBL-harboring organisms have added combination therapy with aztreonam and ceftazidime-avibactam, using ceftazidime-avibactam as a source of the β-lactamase inhibitor avibactam. Current laboratory testing options for the combination of aztreonam-avibactam are limited to broth microdilution (BMD) and broth disk elution (BDE) methods, which are not practical in most clinical laboratories. In this study, we evaluated the performance of aztreonam/avibactam gradient strips on 103 MBL-producing Enterobacterales patient isolates as well as an additional 31 isolates from the CDC AR Bank. All MBL Enterobacterales patient isolates included in this study harbored a New Delhi metallo-β-lactamase (blaNDM) gene. Essential agreement of gradient strip minimal inhibitory concentrations (MICs) for patient isolates compared to BMD was 93.2%. While there are no established breakpoints for aztreonam-avibactam, category agreement (CA) for patient isolates was 97.1% when using the CLSI aztreonam breakpoints. There were no major or very major errors observed. There were three minor errors. Precision for aztreonam-avibactam gradient strip diffusion was 100%. These data demonstrate that the use of gradient strip diffusion for aztreonam-avibactam MIC determination in MBL-producing Enterobacterales is a viable option for clinical laboratories.

Label-free single-cell antimicrobial susceptibility testing in droplets with concentration gradient generation.

Bacterial communities exhibit significant heterogeneity, resulting in the emergence of specialized phenotypes that can withstand antibiotic exposure. Unfortunately, the existence of subpopulations resistant to antibiotics often goes unnoticed during treatment initiation. Thus, it is crucial to consider the concept of single-cell antibiotic susceptibility testing (AST) to tackle bacterial infections. Nevertheless, its practical application in clinical settings is hindered by its inability to conduct AST efficiently across a wide range of antibiotics and concentrations. This study introduces a droplet-based microfluidic platform designed for rapid single-cell AST by creating an antibiotic concentration gradient. The advantage of a microfluidic platform is achieved by executing bacteria and antibiotic mixing, cell encapsulation, incubation, and enumeration of bacteria in a seamless workflow, facilitating susceptibility testing of each antibiotic. Firstly, we demonstrate the rapid determination of minimum inhibitory concentration (MIC) of several antibiotics with Gram-negative E. coli and Gram-positive S. aureus, which enables us to bypass the time-consuming bacteria cultivation, speeding up the AST in 3 h from 1 to 2 days of conventional methods. Additionally, we assess 10 clinical isolates including methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Staphylococcus aureus (MDRSA) against clinically important antibiotics for analyzing the MIC, compared to the gold standard AST method from the United States Clinical and Laboratory Standards Institute (CLSI), which becomes available only after 48 h. Furthermore, by monitoring single cells within individual droplets, we have found a spectrum of resistance levels among genetically identical cells, revealing phenotypic heterogeneity within isogenic populations. This discovery not only advances clinical diagnostics and treatment strategies but also significantly contributes to the field of antibiotic stewardship, underlining the importance of our approach in addressing bacterial resistance.

Combination of Intracanal Medicines with Zinc Oxide Nanoparticles for Effective Eradication of Enterococcus faecalis

Background: Intracanal-resistant infections are the most important reason for root canal treatment failure. Therefore, cleaning the root canals is crucial. The efficiency of nanomedicines in eliminating intracanal contaminations and preventing bacterial attachment to surfaces are new strategies in this field. This study aimed to evaluate the effect of intracanal medicines, including calcium hydroxide (CH), zinc oxide nanoparticles (nZnO), pantoprazole (PP), and chlorhexidine (CHX) on Enterococcus faecalis (E. faecalis) in the individual and combinatorial form in an in vitro design in order to introduce an effective medicine for the eradication of E. faecalis. Sterilized water and sodium hypochlorite (NaOCl) 5.25% were used as the negative and positive control of antimicrobial activity, respectively. Methods: E. faecalis (ATCC 29212) was used for bacterial assays. The powder and the gel forms of the tested compounds were used in the analysis. The antibacterial and antibiofilm activities of the medicines were evaluated by the Minimum Inhibitory Concentration (MIC) determination using the broth microdilution method. The anti-biofilm effect of the tested compounds was assessed by Minimum Biofilm Inhibitory Concentration (MBIC). The antibacterial and antibiofilm effects of antimicrobial agent combinations were detected using the checkerboard assay and determination of the Fractional Inhibitory Concentration Index (FICI). Time-kill kinetics were conducted at zero, 2, 4, 6, 8, 10, 12, and 24 h. Results: Except for the MIC of the NaOCl, which was significantly lower than CH, there was no statistically significant difference between groups in MIC and MBIC. Only the mixture of CHX and nZnO and the mixture of PP and nZnO had a synergic effect against the planktonic and biofilm form of E. faecalis. Except for CHX and ZnO nanoparticles in gel form, other medicines did not show significant anti-biofilm effects against E. faecalis. Furthermore, the gel for CHX and nZnO showed stronger anti-biofilm effects. Conclusion: Combination therapy of common intracanal medicines with zinc oxide nanoparticles would lead to higher antibacterial and anti-biofilm effects against E. faecalis compared to conventional intracanal medicaments.

Highly efficient and broad-spectrum antibacterial carbon dots combat antibiotic resistance

Bacterial infections have become a major global public health issue, particularly with the emergence of multidrug-resistant strains. Therefore, developing non-antibiotic antimicrobial agents is crucial for treating drug-resistant bacterial infections. Building on previous research into natural products as novel antibacterial agents, this study synthesized curcumin-derived carbon dots using curcumin and ethylenediamine as raw materials through a hydrothermal method. The resulting carbon dots not only improved the water solubility and stability of curcumin but also exhibited highly efficient broad-spectrum antibacterial activity. Detailed investigations into the antibacterial performance and mechanisms of the carbon dots were conducted through experiments such as minimum inhibitory concentration (MIC) determination, live/dead bacterial staining, morphological studies, nucleic acid concentration detection, and reactive oxygen species (ROS) detection. The results indicated that the carbon dots significantly damaged the structural integrity of bacteria and generated large amounts of ROS. They exhibited remarkable antibacterial effects against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus, and effectively inhibited drug-resistant MRSA. Their antibacterial efficacy was notably superior to that of broad-spectrum antibiotics such as chloramphenicol and Sulfadiazine. This study highlights the potential application of curcumin-derived carbon dots in combating bacterial infections and provides valuable insights for developing novel antibacterial agents derived from natural products.

Assessment of the potential susceptibility of planktonic cells and bacterial biofilms by diosgenin

Bacterial biofilms are formed by a complex community of microorganisms, held together by a matrix that provides structural support and stability to microbial communities, making infections associated with biofilms more difficult to treat. In the face of growing research in the health area in search of antibiotics with action against biofilms, natural products present an effective alternative strategy. In this context, the aim of this study was to evaluate bacterial biofilm formation, assess the susceptibility of planktonic cells to diosgenin, and examine its efficacy on inhibiting and treating biofilms. The broth microdilution method was used to determine the Minimum Inhibitory Concentration (MIC). Six bacterial strains were tested: Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 9027, Pseudomonas aeruginosa 24, Enterococcus faecalis INCQS 0017, Enterococcus faecalis INCQS 00018, and Enterococcus faecalis INCQS 654. The crystal violet method was employed to evaluate the potential of diosgenin in biofilm inhibition and eradication. Chlorhexidine gluconate served as a reference standard for the antibiofilm assays, along with the antibiotics norfloxacin, ampicillin, and gentamicin. MIC determination revealed that diosgenin exhibited intrinsic antimicrobial activity, with an MIC of 406 μg/mL against S. aureus ATCC 25923. The antibiofilm activity showed significant inhibition for E. faecalis 17 and moderate inhibition against E. faecalis 654 and S. aureus 25923. Furthermore, diosgenin effectively eradicated preformed biofilms of S. aureus 25923 and P. aeruginosa 9027, indicating its ability to interfere with biofilm biomass. These results suggest that diosgenin not only inhibits bacterial biofilm formation but also eradicates established biofilms, highlighting it as a promising candidate for the development of products aimed at combating bacterial biofilms.

Synthesis and evaluation of novel sulfamethoxazole-containing 1,8-dioxo-decahydroacridine derivatives: Antimicrobial activity and emission properties

A straightforward and efficient method for synthesizing 1,8-dioxo-decahydroacridine derivatives is presented, utilizing a one-pot, three-component condensation reaction involving dimedone, sulfamethoxazole, and aromatic aldehydes. The primary objective of this synthesis is to incorporate the sulfamethoxazole ring into the acridine family structure and examine the antibacterial effects and fluorescence properties of the compounds. The antibacterial activity of the synthesized compounds was evaluated against various gram-positive and gram-negative microorganisms, as well as a fungal strain, using the disk diffusion method, minimum inhibitory concentration determination, and bactericidal assays. Notably, five compounds exhibited potent antibacterial activity against both strains of Bacillus cereus. Furthermore, the influence of pH on the fluorescence emission intensity and absorption spectrum of the synthesized compounds was investigated, revealing that all products displayed fluorescence radiation.

EFFECT OF MORINGA (Moringa oleifera) LEAF EXTRACT ON IN VITRO INHIBITION OF Candida albicans BIOFILM

Background: Oral candidiasis is an opportunistic infection in the oral cavity. This infection is caused by the fungus Candida albicans. The most important virulent attribute of this fungus is its ability to form biofilms, which can adhere to mucosa, epithelial lining, organs, prostheses or dentures. The formed biofilm is resistant to antifungal drugs. Objectives: The purpose of this study was to determine the effect of Moringa (Moringa oleifera) leaf extract on the inhibition of Candida albicans biofilm formation in vitro. Methods: This type of research is experimental study using post tests with control group design. Fungus used in this study was Candida albicans ATCC 10231. Determination of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were conducted by microdilution method. Inhibition test of C. albicans biofilm formation was carried out using the polystyrene microplate assay method. Concentration of extracts used in this study were 12.5%, 6.25%, 3.13% and 1.57%. Inhibitory activity of biofilm was measured using a crystal violet (CV) assay. Results: The results showed that MIC of Moringa leaf extract against the fungus C. albicans was at a concentration of 6.25%. Moringa leaf extract starting at a concentration of 3.13% already has had ability to inhibit formation of C. albicans biofilm. Conclusion: It can be concluded that Moringa leaf extract can inhibit formation of Candida albicans biofilm, so it can be developed as an alternative herbal ingredient to prevent oral candidiasis.EFFECT OF MORINGA (Moringa oleifera) LEAF EXTRACT ON IN VITRO INHIBITION OF Candida albicans BIOFILM

The antibiotic resistance profiles of Pseudomonas aeruginosa in the Asia Cornea Society Infectious Keratitis Study

PurposeTo describe the prevalence and antibiotic resistance profiles of Pseudomonasaeruginosa isolated from the Asia Cornea Society Infectious Keratitis Study (ACSIKS).MethodsAll bacterial isolates from ACSIKS underwent repeat microbiological identification in a central repository in Singapore. Minimum inhibitory concentration (MIC) determination was conducted for isolates of P. aeruginosa against thirteen antibiotics from 6 different classes, and categorized based on Clinical Laboratory Standard Institutes’ reference ranges. The percentage rates of resistance (non-susceptibility) to each antibiotic included isolates of both intermediate and complete resistance. Multi-drug resistance (MDR) was defined as non-susceptibility to at least one agent in three or more antimicrobial classes.ResultsOf the 1493 unique bacterial specimens obtained from ACSIKS, 319 isolates were of P. aeruginosa. The majority of isolates were from centers in India (n = 118, 37%), Singapore (n = 90, 28.2%), Hong Kong (n = 31, 9.7%) and Thailand (n = 30, 9.4%). The cumulative antibiotic resistance rate was the greatest for polymyxin B (100%), ciprofloxacin (17.6%) and moxifloxacin (16.9%), and lowest for cefepime (11.6%) and amikacin (13.5%). Isolates from India demonstrated the highest antibiotic resistance rates of all the centers, and included moxifloxacin (47.5%) and ciprofloxacin (39.8%). Forty-eight of the 59 MDR isolates also originated from India. Antibiotic resistance rates were significantly lower in the other ACSIKS centers, and were typically less than 10%.ConclusionsThe antibiotic resistance profiles of P. aeruginosa varied between different countries. While it was low for most countries, substantial antibiotic resistance and a significant number of multi-drug resistant isolates were noted in the centers from India.

Assessing chemical composition and biological activity in Eucryphia cordifolia Cav. (Cunoniaceae): A study applying the ecological niche centrality hypothesis

Eucryphia cordifolia Cav. native to the forests of southern Chile, was used for medicinal purposes by the Mapuche people in ancient times. However, there are no recent studies on the biological properties of its leaves in their natural habitat. This study assessed how the location of E. cordifolia populations in their ecological niche (center or periphery) impacts the chemical composition, antioxidant activity, and antimicrobial properties of leaf extracts.Ultra-Liquid Chromatography with Diode Array Detector, coupled to Quadrupole-Time of Fly Mass Spectrometry was used to characterize the chemical composition of the extracts (UHPLC-DAD-QTOF). The antioxidant activity was assessed through DPPH, ABTS, and ORAC assays, and the total polyphenol content was determined using the Folin-Ciocalteu method. Antimicrobial activity was analyzed using methods such as agar diffusion, determination of minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and biofilm formation assay. The leaves extract contained 28 identified compounds, including flavonols, flavones, flavanols, and other bioactive compounds. The peripheral population exhibited higher antioxidant values based on ABTS and ORAC methods in autumn and spring, and higher total polyphenol levels only in autumn. The E. cordifolia extracts significantly inhibited the growth of E. coli, S. aureus, and P. aeruginosa, with minimum bactericidal concentration values ranging between 1.2 and 5.3 mg/mL, lower than in other studies. Additionally, the peripheral population displayed greater inhibition of C. glabrata biofilm formation compared to the centroid and intermediate populations in both summer and autumn. In conclusion, this first comprehensive investigation of E. cordifolia reveals that its leaves are a valuable source of bioactive compounds with biotechnological potential.

Evaluation of the Reveal® AST (SPECIFIC) for Antimicrobial Susceptibility Testing from Positive Blood Culture Spiked with Carbapenem-Resistant Isolates.

As bloodstream infections and associated septic shock are common causes of mortality in hospitals, rapid antibiotic susceptibility testing (AST) performed directly on positive blood cultures is needed to implement an efficient therapy in clinical settings. We evaluated the Reveal® rapid AST system on a collection of 197 fully characterized carbapenem-resistant Enterobacterales, including 177 carbapenemase producers (CPE) spiked in blood culture bottles. The clinical categorization based on the Minimal Inhibitory Concentration (MIC) determination of eighteen antimicrobial molecules was compared to the clinical categorization based on the disk diffusion assay as a reference. The Reveal AST system provided results within a mean time to result of 5 h. Overall, the categorical agreement (CA) between the two techniques was 94.1%. The rates of very major errors (VMEs), major errors (MEs) and minor errors (mEs) were 3.8%, 3.7% and 5.6%, respectively. Imipenem was the antimicrobial with the lowest CA rate (78.7%), with rates of 15% VMEs and 10.7% MEs, but the performances were better when considering only the non-CPE category (CA of 89%). On this resistant collection of Enterobacterales with numerous acquired β-lactamases, the Specific Reveal assay proved to be useful for a rapid determination of AST compatible with a quick adaptation of the patient's antimicrobial treatment.

Tigecycline Resistant Pattern among Carbapenem Resistant Gram-negative Bacilli: Prospective Cross-sectional Study

Introduction: Tigecycline is a unique tetracycline class of semi-synthetic, last-line broad spectrum antibiotic against multi-drug-resistant bacteria. However, recently, resistance to this antibiotic is on the rise. Aims: This study was conducted to determine the prevalence of tigecycline resistance amongst carbapenem-resistant Gram-negative bacilli (GNB) isolated from clinical samples (pus and sputum) as well as to evaluate their antimicrobial susceptibility pattern. Study design: Prospective cross sectional Place and Duration of Study: Department of Microbiology at Sanjay Gandhi Post Graduate Institute of Medical Sciences Lucknow, between January 2023 and December 2023. Methodology: Identification of GNB grown on culture was done by conventional biochemical tests and later validated by MALDI-TOF MS. The antimicrobial sensitivity testing of isolates was done using the E-test, and disk diffusion method. Minimum inhibitory concentration determination was done by Broth micro-dilution (BMD) method. Results: Amongst 8326 pus and respiratory samples, GNBs were recovered from 63.15% (5258/8326). Of 5258 GNB isolates, 50.74% (2668) were carbapenem-resistant, while 7.85% (413) demonstrated resistance to both tigecycline and carbapenem. Common isolates in this group were Klebsiella pneumoniae (37.04%), Acinetobacter spp. (25.18%), Enterobacter spp. (14.28%) and Escherichia coli (12.59%). BMD results demonstrated highest activity of tigecycline against carbapenem-resistant E. coli, followed by Citrobacter and Enterobacter. It works against resistant strains of Acinetobacter baumannii and K. pneumoniae as well, but in higher concentrations. Conclusion: High tigecycline resistance (one of the last-resort drugs) among carbapenem resistant GNB isolates is a matter of clinical concern, leaving physicians with limited options for treatment of such infections. Proper adherence to the policies of antimicrobial stewardship programs can reduce the emergence of resistance.

Identification of vancomycin resistance in enterococcus & determination of minimal inhibitory concentration of vancomycin by E-test

Infections due to Enterococcal species is 2 prevalent cause of Nosocomial Infections in health care settings. They are crucial because to their inherent resistance to penicillins, cephalosporins, and low level aminoglycosides & their tendency to develop higher level of aminoglycoside and vancomycin resistance, which leaves few therapeutic alternatives for therapy. Isolation and identification of Enterococci with their Antibiogram & determination of Minimal Inhibitory Concentration (MIC) of Vancomycin by E-test. The antibiogram generated by Kirby Baur disc diffusion method and standard microbiological procedures were used to identify enterococcal isolates from distinct clinical samples. MIC level for Vancomycin was tested for all the isolates using E-test from BIOMERIEUX. Total of 108 isolates (62 E.faecalis and 46 E. faecium) were tested by E-test for MIC of Vancomycin, of which 40 (37.03%) (22 E.faecalis & 18 E. faecium) were Vancomycin Resistant (VRE) strains, having MIC level >32 μg/ml. Nine strains out of 40 (45%) were found to have MIC >256 μg/ml. The enterococcal isolates were observed to be sensitive to Linezolid (80%), Ofloxacin (74%), Amoxyclav (74%) and Cefaperazone (70%). For the Indian medical community, the rise of antibiotic resistance among Enterococcal species has been a significant concern. So this is need of present era that our laboratories must be well-equipped to report the resistance patterns displayed by enterococci in order to practise evidence-based medicine and reduce hospital acquired infections.

Skipped wells and scientific error during fosfomycin agar dilution and broth microdilution lead to inconsistent minimal inhibitory concentrations and may be cause for reevaluating testing methods for Klebsiella pneumoniae.

Despite the first-line recommendation of fosfomycin for uncomplicated urinary tract infections (UTIs), there are pressing barriers for optimizing its use for the treatment of non-Escherichia coli Enterobacterales UTI. There are no approved breakpoints for oral use against other Enterobacterales, and the recommended agar dilution (AD) reference method for minimal inhibitory concentration (MIC) determination is largely impractical. Using 160 clinical Klebsiella pneumoniae isolates, we sought to understand rates of skipped wells and MIC imprecision in broth microdilution (BMD) and how that compares to rates of error using AD. Though the Clinical and Laboratory Standards Institute refers to the skipped well phenomena in their recommendation against the use of BMD, there is a paucity of data on its frequency. While AD and BMD produced similar MIC50/90 values (32/256 µg/mL for AD and 64/256 µg/mL for BMD), essential agreement was poor. No-growth wells at concentrations below the MIC occurred in up to 10.9% of wells at a given concentration, as the most frequent scientific error. Growth in concentrations above the measured MIC occurred in up to 3.3% of wells and was seen within three dilutions of the MIC for BMD. Observation of single colonies either at or beyond the measured MIC for AD was also common and occurred up to 8.3% and 2.5% of the time, respectively. The frequent scientific error in both testing methods should prompt re-evaluation of AD guidelines and expansion of MIC testing methods for fosfomycin susceptibility testing, as poor agreement with another method prone to scientific error should not be the main detractor from BMD use.IMPORTANCEDespite the recommendation of fosfomycin for uncomplicated urinary tract infections (UTIs), there are barriers for optimizing its use. There are no approved breakpoints for oral use against other Enterobacterales, and the recommended agar dilution (AD) reference method for MIC determination is largely impractical. The use of broth microdilution (BMD) for fosfomycin testing is not recommended by the Clinical and Laboratory Standards Institute due to unsatisfactory precision and skipped wells-occurrence of no-growth in a single well before the minimal inhibitory concentration (MIC)-and trailing endpoints. We sought to understand rates of skipped wells and growth at concentrations above measured MICs in BMD and how that compares to scientific error using AD. No-growth wells at concentrations below the MIC occurred in up to 10.9% of wells for BMD and single colonies at or beyond measured MICs for AD were also common. Frequent scientific error in both methods should prompt re-evaluation of both AD and BMD for fosfomycin susceptibility testing.

Determination of Minimum Inhibitory Concentrations of Sophorolipids Against Selected Halotolerant and Moderately Halophilic Bacteria, and Extremely Halophilic Archaea Isolated from Salt Samples and Salted Skins

Determination of Minimum Inhibitory Concentrations of Sophorolipids Against Selected Halotolerant and Moderately Halophilic Bacteria, and Extremely Halophilic Archaea Isolated from Salt Samples and Salted Skins