Search Results (5)

Methicillin-resistant Staphylococcus aureus (MRSA) continues to represent a significant clinical challenge, characterized by consistently elevated rates of morbidity and mortality. Care regimen success is still difficult and necessitates assessing new antibiotics as well as supplemental services, including source control and searching for alternative approaches to combating it. Hence, we propose to synthesize silver nanoparticles (Ag-NPs) by employing a cell-free filter (CFF) of Streptomyces sp. to augment antibiotic activity and combat biofilm-forming MRSA. Seven bacterial isolates from clinical samples were identified, antibiotics were profiled with Vitek-2, and the phenotypic detecting of biofilm with Congo red medium and microplate assay was carried out. The PCR technique was used for detecting genes (icaA and icaD) coded in biofilm forming. The characterization of Ag-NPs was performed using several analytical methods, such as UV spectroscopy, dynamic light scattering (DLS), zeta potential measurement, transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The antibacterial properties of Ag-NPs and oxacillin–Ag-NPs were assessed against standard strains and clinical isolates by employing the agar well diffusion technique and the microdilution assay. The biogenic synthesis Ag-NPs resulted in uniformly spherical particles, with an average size of 20 nm. These Ag-NPs demonstrated significant activity against biofilm-forming MRSA, with minimum inhibitory concentrations (MICs) ranging from 12 to 15 μg/mL. Additionally, Ag-NPs completely impede biofilm formation by MRSA at sublethal doses of 0.75 MICs. The expression levels of the icaA and icaD genes were reduced by 1.9- to 2.2- and 2.4- to 2.8-fold, respectively. A significant synergistic effect was noted when Ag-NPs were used in combination with oxacillin, leading to reduced MICs of 1.87 μg/mL for oxacillin and 4.0 μg/mL for Ag-NPs against MRSA. The FICi of 0.375 further validated the synergistic relationship between oxacillin and Ag-NPs at the concentrations of 1.87 and 4 μg/mL. Findings from the time-kill test demonstrated the highest reduction in log₁₀ (CFU)/mL of the initial MRSA inoculum after 12-hour exposure. The cytotoxicity analysis of Ag-NPs revealed no significant cytotoxic effects on the human skin cell line HFB-4 at low concentrations, with IC₅₀ values of 61.40 µg/mL for HFB-4 and 34.2 µg/mL for HepG-2. Comparable with oxacillin–Ag-NPs, Ag-NPs showed no cytotoxic effects on HFB-4 at different concentrations and exhibited an IC₅₀ value of 31.2 against HepG-2-cells. In conclusion, the biosynthesis of Ag-NPs has demonstrated effective antibacterial activity against MRSA and has completely hindered biofilm formation, suggesting a valuable alternative for clinical applications. Full article

Background/Objectives: The International Organization for Standardization (ISO) 20776-2:2021, which replaces ISO 20776-2:2007, focuses solely on the performance of antimicrobial susceptibility testing (AST) assays, emphasizing the ISO 20776-1 broth microdilution method as the reference standard. Consequently, categorical agreement (CA) and associated errors should not be applied. We verified the Vitek Reveal AST assay according to both ISO 20776-2:2021 and ISO 20776-2:2007 criteria. Methods: Samples from 100 simulated and clinical Gram-negative (GN) positive blood cultures (PBCs) were tested at a large teaching hospital. The simulated GN-PBCs were obtained from a hospital collection of isolates selected to represent diverse antimicrobial resistance profiles. The Reveal assay results were compared with those from the reference assay, and the time to result (TTR) for the Reveal assay was calculated. Results: The essential agreement rates were 96.1% (816/849) for simulated and 98.8% (929/940) for clinical GN-PBC samples. The bias values were −3.1 for simulated and −11.0 for clinical samples. The CA rates were 97.7% (808/827) for simulated and 99.2% (924/931) for clinical samples. The mean TTR ± SD (hours) for resistant organisms was significantly lower (4.40 ± 1.15) than that for susceptible, increased exposure (5.52 ± 0.48) and susceptible (5.54 ± 0.49) organisms. Conclusions: Our findings reinforce the potential of the Reveal assay as a valuable tool and support its implementation in clinical microbiology laboratories. Full article

Candida auris was reported by the WHO as second to Cryptococcus neoformans, in the list of nineteen fungal priority pathogens, along with two species with a new nomenclature, Nakaseomyces glabrata (Candida glabrata) and Pichia kudriavzevii (Candida krusei). This novel classification was based on antifungal resistance, the number of deaths, evidence-based treatment, access to diagnostics, annual incidence, and complications and sequelae. We assessed which molecular assays have been used to diagnose Candida auris outbreaks in the last five years. Using “Candida auris; outbreak; molecular detection” as keywords, our search in PubMed revealed 32 results, from which we selected 23 original papers published in 2019–2024. The analyzed studies revealed that the detection methods were very different: from the VITEK® 2 System to MALDI TOF (Matrix-Assisted Laser Desorption Ionization–Time of Flight), NGS (Next-Generation Sequencing), WGS (Whole Genome Sequencing), and commercially available real-time PCR (Polymerase Chain Reaction) assays. Moreover, we identified studies that detected antifungal resistance genes (e.g., FKS for echinocandins and ERG11 for azoles). The analyzed outbreaks were from all continents, which confirms the capability of this yeast to spread between humans and to contaminate the environment. It is important that real-time PCR assays were developed for accurate and affordable detection by all laboratories, including the detection of antifungal resistance genes. This will allow the fast and efficient implementation of stewardship programs in hospitals. Full article

Due to the misuse and overuse of antibiotics, antibiotic residues accumulate in natural environments, leading to the development of antibiotic-resistant bacteria (ARBs). The presence of ARBs in bodies of water poses health hazards to the surrounding community. This study focused on Laguna Lake, the largest lake in the Philippines, which serves as a water source for agriculture and domestic purposes. We aimed to detect the presence of antibiotic-resistant Escherichia coli from the lake waters and potential reservoirs of resistance as well as determine the multiple antibiotic resistance (MAR) indices of the isolates. E. coli (n = 450) was isolated from fecal-associated samples (chicken, cow, pig, human, sewage) and water samples (sites in Laguna Lake and selected river tributaries). The isolates were subjected to an antibiotic resistance assay using VITEK 2^®. Among the 16 antibiotics tested, the isolates exhibited varying resistance to 14, but complete susceptibility to amikacin and tigecycline was observed. Isolates were most frequently resistant to ampicillin (196/450, 43.6%). Among fecal-associated samples, chicken isolates exhibited the highest MAR index (0.174), whereas samples from Pila River exhibited the highest MAR index (0.152) among water samples. The results of this study demonstrate the presence of multidrug-resistant E. coli in samples collected around Laguna Lake and reveal fecal and sewage sources as potential reservoirs of ARBs in the water body. With this information, the public is urged to use antibiotics responsibly to help mitigate the spread of antibiotic resistance. Full article

Streptococcus agalactiae is responsible for serious infections in newborn babies, pregnant women, and other patients. The aim of this study was to evaluate antimicrobial susceptibility, serotype distribution, and virulence determinants of the S. agalactiae isolates derived from clinical specimens considering the global increase of both antibiotic resistance and virulence. A total of 165 isolates were identified and serotyped by PCR techniques. Antimicrobial susceptibility was assessed by disk diffusion method, gradient diffusion method and VITEK^® System. Virulence associated genes were investigated by PCR; ability to form biofilm was assessed using a microtiter plate assay. The highest observed MIC value for penicillin G was 0.12 µg/mL, seen in 8.5% of isolates. Resistance to erythromycin and clindamycin were found in 30.38% and 24.8% of the strains, respectively. The serotype III (32.73%), V (25.45%), and Ia (18.18%) were found as the most frequently represented. Previously unidentified strains in Poland, belonging to serotypes VI (three strains) and VII (one strain) were recognized. The presence of genes encoding various virulence factors as well as diverse ability to form biofilm were found. In conclusion, macrolide-resistance and decreased susceptibility to penicillin G were revealed signifying the increasing resistance among group B streptococci. Moreover, the presence of genes encoding various virulence factors and the ability to form biofilm were confirmed indicating their role in the pathomechanisms of the evaluated GBS infections. Full article