Vancomycin-resistant S. Aureus Research Articles

Phenotypic and genotypic characterization of methicillin and vancomycin-resistant Staphylococcus aureus isolated from human clinical samples in Kerman hospitals, Iran

Today, methicillin-resistant S. aureus (MRSA) isolates are widely spread in both communities and hospitals, causing high morbidity and mortality worldwide. However, the emergence of vancomycin resistance among MRSA isolates has been perceived as a formidable threat to therapeutic management. Therefore, the present study aimed to determine the frequency and antibiotic susceptibility of MRSA and vancomycin-resistant S. aureus (VRSA) strains in Kerman hospitals in Iran. A total of 455 human clinical samples were collected from Kerman hospitals. The samples were examined for the presence of S. aureus and the isolates were tested for their susceptibility to antibiotics by disk diffusion method. MRSA and VRSA isolates were detected by a combination of phenotypic and genotypic methods. Sixty-five coagulase-positive S. aureus strains were isolated, and the higher isolation rate was from wound samples (47.7%) compared with blood (18.5%), respiratory tract secretions (10.8%), urine (10.8%), body fluids (9.2%), abscess (1.5%), and cerebrospinal fluids (1.5%). The isolates exhibited high resistance toward tetracycline (69.2%). Thirty-four isolates (52.3%) were categorized as MRSA (phenotypically resistant to cefoxitin). mecA gene was amplified in 97.1% of MRSA strains. Moreover, only one MRSA strain was resistant to vancomycin (MIC=128), and this isolate carried the vanA gene. The frequency of MRSA was not significantly associated with gender, age, sample type, and sampling locality (p>0.05). Given the alarming rate of resistance among MRSA isolates, monitoring of antibiotic resistance should be performed to reduce treatment failure in patients with staphylococcal infections. Although vancomycin remains a drug of choice for MRSA, our study suggests that its efficacy may be limited by resistance development.

A hope for ineffective antibiotics to return to treatment: investigating the anti-biofilm potential of melittin alone and in combination with penicillin and oxacillin against multidrug resistant-MRSA and -VRSA.

The emergence and rapid spread of multi-drug resistant (MDR) bacterial strains, such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant S. aureus (VRSA), have posed a significant challenge to the medical community due to their ability to form biofilm and develop resistance to common antibiotics. Traditional antibiotics that were once effective in treating bacterial infections are now becoming increasingly ineffective, leading to severe consequences for patient outcomes. This concerning situation has called for urgent research to explore alternative treatment strategies. Recent studies have shown that antimicrobial peptides (AMPs) hold promise as effective agents against biofilm-associated drug-resistant infections as well as to enhance the efficacy of conventional antibiotics. Accordingly, we aimed to investigate the antimicrobial and antibiofilm effects of melittin AMP, both alone and in combination with penicillin and oxacillin, against biofilm-forming MDR-MRSA and -VRSA. In this study, we investigated the kinetics of biofilm formation and assessed various parameters related to the antimicrobial and antibiofilm efficacy of melittin and antibiotics, both alone and in combination, against MDR-MRSA and -VRSA. The antimicrobial parameters included the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), Fractional Inhibitory Concentration Index (FICi), Fractional Bactericidal Concentration Index (FBCi), and the antibiofilm activity of melittin and antibiotics indicated by the Minimum Biofilm Inhibitory Concentration (MBIC), Minimal Biofilm Eradication Concentration (MBEC), Fractional Biofilm Inhibitory Concentration Index (FBICi), and Fractional Biofilm Eradication Concentration Index (FBECi). The MIC results showed that all S. aureus isolates were resistant to penicillin (≥0.25 μg/mL), and 66% of isolates were resistant to oxacillin. The geometric means of the MIC values for penicillin, oxacillin, and melittin were 19.02, 16, and 1.62 μg/ml, respectively, and the geometric means of the MBC values for penicillin, oxacillin, and melittin were 107.63, 49.35, and 5.45 μg/ml, respectively. The study revealed that the combination indexes of melittin-penicillin and melittin-oxacillin, as determined by FIC values against all isolates, were 0.37 and 0.03, respectively. Additionally, melittin-penicillin and melittin-oxacillin exhibited combination indexes based on FBC values against all isolates at 1.145 and 0.711, respectively. Besides, melittin inhibited the biofilm formation of all S. aureus isolates, with MBIC values ranging from 10 to 1.25 μg/mL, and MBEC values ranging from 40 to 10 μg/mL. Generally, the combination indexes of melittin-penicillin and melittin-oxacillin, determined using FBIC values against all isolates, were 0.23 and 0.177, respectively. Moreover, melittin-penicillin and melittin-oxacillin typically had combination indexes based on FBEC values against all isolates at 5 and 2.97, respectively. In conclusion, our study provides evidence that melittin is effective against both planktonik and biofilm forms of MRSA and VRSA and exhibits significant synergistic effects when combined with antibiotics. These results suggest that melittin and antibiotics could be a potential candidate for further investigation for in vivo infections caused by MDR S. aureus. Furthermore, melittin has the potential to restore the efficacy of penicillin and oxacillin antibiotics in the treatment of MDR infections. Applying AMPs, like melittin, to revive beta-lactam antibiotics against MRSA and VRSA is an innovative approach against antibiotic-resistant bacteria. Further research is needed to optimize dosage and understand melittin mechanism and interactions with beta-lactam antibiotics for successful clinical applications.

Determination of Vancomycin Minimum Inhibitory Concentration in Methicillin-resistant Staphylococcus Aureus by Various Phenotypic Methods: A Cross-sectional Study

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) has become a global threat, causing nosocomial and community-acquired infections, thus significantly contributing to morbidity and mortality. These organisms often exhibit Multidrug Resistance (MDR) to various groups of antimicrobials. The emergence of reduced susceptibility to vancomycin and resistance further exacerbates the problem, leaving clinicians with few therapeutic options to treat serious, life-threatening infections. Aim: To determine the vancomycin susceptibility of MRSA using various phenotypic methods. Materials and Methods: The present cross-sectional study was conducted in the Department of Microbiology, SHKM Government Medical College, Nalhar, Haryana, India, from February 2019 to January 2020. All MRSA isolates were included, totaling 66 MRSA isolates. MRSA isolates were screened using the cefoxitin disc (30 μg) according to the Clinical and Laboratory Standards Institute (CLSI) recommended disc diffusion method. The Minimum Inhibitory Concentration (MIC) of vancomycin was determined using phenotypic methods such as broth dilution, agar dilution and Epsilometer test (E-test). Demographic details such as age, sex, site of the sample and diagnosis were recorded. Statistical analysis was performed using the Pearson’s product-moment correlation. The significance level was set at a probability of <0.05 with a confidence interval of 95%. Results: A total of 66 (30.3%) isolates of Staphylococcus aureus (S. aureus) were identified as MRSA using standard laboratory tests and were studied for vancomycin MIC using phenotypic methods: Broth dilution, agar dilution and E-test. Vancomycin- intermediate S. aureus (VISA) was observed in 54.5%, 42.4% and 51.5% of MRSA by E-test, agar dilution and broth dilution, respectively. The presence of Vancomycin-resistant S. aureus (VRSA) was not detected in the region. When compared with broth dilution, E-test results demonstrated higher accuracy (97%) than agar dilution (90.9%). Conclusion: The present study highlights the emergence of VISA in more than 40% of the MRSA in the region. The higher MIC of vancomycin emphasises the need for strict adherence to antibiotic prescription guidelines to prevent further emergence of vancomycin resistance.

Synthesis of 1,2,3-triazole-piperazin-benzo[b][1,4]thiazine 1,1-dioxides: antibacterial, hemolytic and in silico TLR4 protein inhibitory activities.

In this study, we designed and synthesized a number of novel 1,2,3-triazole-piperazin-benzo[b][1,4]thiazine 1,1-dioxide derivatives and investigated their in vitro antibacterial and hemolytic activity. When compared to the lead chemical, dicloxacillin, the majority of the compounds demonstrated acceptable activity. Among them, the most promising compounds 6e, 6g, 6i, 8d, and 8e exhibited excellent antibacterial activity against the methicillin-susceptible S. aureus (MSSA), methicillin-resistant S. aureus (MRSA), and vancomycin-resistant S. aureus (VRSA) with MIC values of 1.56 ± 0.22 to 12.5 ± 1.75 μg mL-1, respectively, The percentage of hemolysis ranged from 21.3 μg mL-1 to 33.8 μg mL-1. Out of the six compounds (6i, 6e, 6f, 6g, 8e, 8d) tested compound 8e and 8d displayed minimal or negligible hemolytic activity across all the tested concentrations 29.6% and 30.2% recorded at 100 μg mL-1 concentration respectively. In silico docking studies were performed to evaluate the molecular interactions of 6e, 6f, 6g, 6i, 8d, and 8e compounds with Human, Mouse and Bovine TLR4 proteins (PDB: 3FXI, 3VQ1, 3RG1) and observed that three of the compounds (6i, 8d, and 8i) had appreciable binding energies ranging from -8.5 to -9.0 Kcal mol-1. Finally, the in silico pharmacokinetic profile was predicted for potent compounds 8d, 8e and 6i using SWISS/ADME, All compounds investigated in this study adhered to Lipinski's rule of five with slight deviation in molecular weight (8d and 8e).

Biofilms: A developmental niche for vancomycin-intermediate Staphylococcus aureus

Staphylococcus aureus are gram-positive bacteria responsible for a wide array of diseases, ranging from skin and soft tissue infections to more chronic illnesses such as toxic shock syndrome, osteomyelitis, and endocarditis. Vancomycin is currently one of the most effective antibiotics available in treating patients infected with methicillin-resistant S. aureus (MRSA), however the emergence of vancomycin-resistant S. aureus (VRSA), and more commonly vancomycin-intermediate S. aureus (VISA), threaten the future efficacy of vancomycin. Intermediate resistance to vancomycin occurs due to mutations within the loci of Staphylococcal genes involved in cell wall formation such as rpoB, graS, and yycG. We hypothesized the VISA phenotype may also arise as a result of the natural stress occurring within S. aureus biofilms, and that this phenomenon is mediated by the RecA/SOS response. Wildtype and recA null mutant/lexAG94E strains of S. aureus biofilms were established in biofilm microtiter assays or planktonic cultures with or without the addition of sub-inhibitory concentrations of vancomycin (0.063 mg/l - 0.25 mg/L ciprofloxacin, 0.5 mg/l vancomycin). Efficiency of plating techniques were used to quantify the subpopulation of biofilm-derived S. aureus cells that developed vancomycin-intermediate resistance. The results indicated that a greater subpopulation of cells from wildtype biofilms (4.16 × 102 CFUs) emerged from intermediate-resistant concentrations of vancomycin (4 μg/ml) compared with the planktonic counterpart (1.53 × 101 CFUs). Wildtype biofilms (4.16 × 102 CFUs) also exhibited greater resistance to intermediate-resistant concentrations of vancomycin compared with strains deficient in the recA null mutant (8.15 × 101 CFUs) and lexA genes (8.00 × 101 CFUs). While the VISA phenotype would be an unintended consequence of genetic diversity and potentially gene transfer in the biofilm setting, it demonstrates that mutations occurring within biofilms allow for S. aureus to adapt to new environments, including the presence of widely used antibiotics.

Methicillin-Resistant Staphylococcus aureus and Coagulase-Negative Staphylococcus from School Dining Rooms in Argentina.

Methicillin-resistant Staphylococcus aureus (MRSA) constitutes an important cause for concern in the field of public health, and the role of the food chain in the transmission of this pathogen and in antimicrobial resistance (AMR) has not yet been defined. The objectives of this work were to isolate and characterize coagulase-positive Staphylococcus (CoPS) and coagulase-negative Staphylococcus (CoNS), particularly S. aureus, from school dining rooms located in Argentina. From 95 samples that were obtained from handlers, inert surfaces, food, and air in 10 establishments, 30 Staphylococcus strains were isolated. Four isolates were S. aureus, and the remaining ones (N = 26) belonged to 11 coagulase-negative species (CoNS). The isolates were tested for susceptibility to nine antibiotics. The presence of genes encoding toxins (luk-PV, sea, seb, sec, sed, and see), adhesins (icaA, icaD), and genes that confer resistance to methicillin (mecA) and vancomycin (vanA) was investigated. The resistance rates measured for penicillin, cefoxitin, gentamicin, vancomycin, erythromycin, clindamycin, levofloxacin, trimethoprim-sulfamethoxazole, and tetracycline were 73%, 30%, 13%, 3%, 33%, 17%, 13%, 7%, and 7% of the isolates, respectively. Seventeen AMR profiles were detected, and 11 isolates were multidrug resistant (MDR). Seven methicillin-resistant Staphylococcus isolates were detected in the hands of handlers from four establishments, two of them were MRSA. Two S. aureus isolates presented icaA and icaD, another one, only icaD. The gene vanA was found in two isolates. In relation to S. aureus, resistance to vancomycin but not to gentamicin was detected. School feeding plays a key role in the nutrition of children, and the consumption of food contaminated with MRSA and vancomycin-resistant S. aureus (VRSA) can be a serious threat to health. In particular, it was detected that the handlers were the source of MRSA, VRSA, MR-CoNS (methicillin-resistant coagulase-negative Staphylococcus), and MDR isolates. The results obtained indicate that the vigilance of this pathogen in school dining rooms should be extreme.

Partnering essential oils with antibiotics: proven therapies against bovine Staphylococcus aureus mastitis.

There is an urgent need to develop therapeutic options for biofilm-producing Staphylococcus aureus (S. aureus). Therefore, the renewed interest in essential oils (EOs), especially carvacrol, linalool and eugenol, has attracted the attention of our research group. Multidrug resistance and multivirulence profiles in addition to biofilm production of S. aureus strains isolated from cows with mastitis were evaluated using both phenotypic and genotypic methods. The antimicrobial and antibiofilm activities of EOs were tested using both in vitro and molecular docking studies. Moreover, the interactions between commonly used antibiotics and the tested EOs were detected using the checkerboard method. We found that all our isolates (n= 37) were biofilm methicillin resistant S. aureus (MRSA) producers and 40.5% were vancomycin resistant S. aureus (VRSA). Unfortunately, 73 and 43.2% of the recovered MRSA isolates showed multidrug resistant (MDR) and multivirulence patterns, respectively. The antimicrobial activities of the tested EOs matched with the phenotypic evaluation of the antibiofilm activities and molecular docking studies. Linalool showed the highest antimicrobial and antibiofilm activities, followed by carvacrol and eugenol EOs. Fortunately, synergistic interactions between the investigated EOs and methicillin or vancomycin were detected with fractionalinhibitory concentration index (FICI) values ≤ 0.5. Moreover, the antimicrobial resistance patterns of 13 isolates changed to sensitive phenotypes after treatment with any of the investigated EOs. Treatment failure of bovine mastitis with resistant S. aureus can be avoided by combining the investigated EOs with available antimicrobial drugs. We hope that our findings can be translated into a formulation of new pharmaceutical dosage forms against biofilm-producing S. aureus pathogens.

Identification of Vancomycin Resistance in Methicillin-resistant Staphylococcus aureus in two macaque species and decolonization and long-term prevention of recolonization in Cynomolgus Macaques (Macaca fascicularis).

Methicillin-resistant Staphylococcus aureus (MRSA) is a S. aureus strain with resistance to beta-lactam antibiotics, making it a global human and veterinary health concern. Specifically, immunosuppressed patients have a remarkably higher risk of clinical MRSA infections with significantly increased rates of prolonged clinical recovery, morbidity, and mortality. The current treatment of choice for MRSA is vancomycin. Importantly, we report the first known vancomycin-resistant S. aureus (VRSA) carriers in a cohort of Mauritian cynomolgus macaques (CM) imported to the Oregon National Primate Research Center (ONPRC), with a MRSA carrier rate of 76.9% (10/13 animals). All MRSA isolates also demonstrated resistance to vancomycin with prevalence of vancomycin-intermediate Staphylococcus aureus (VISA) at 30% (3/10 MRSA-positive CMs) and VRSA at 70% (7/10 MRSA-positive CMs). Additionally, we identified VRSA in a rhesus macaque (RM) housed within the same room as the VRSA-positive CMs and identified a MRSA/VISA carrier rate of 18.8% in RMs (3/16 positive for both MRSA and VISA) in unexposed recently assigned animals directly from the ONPRC RM breeding colony. Considering that the MRSA and VRSA/VISA-positive CMs future study aims included significant immunosuppression, MRSA/VRSA/VISA decolonization treatment and expanded "MRSA-free" practices were employed to maintain this status. We report the first controlled study using in-depth analyses with appropriate diagnostic serial testing to definitively show an MRSA decolonization therapy (90% success rate) and expanded barrier practice techniques to successfully prevent recolonization (100%) of a cohort of CMs MRSA-free (up to 529 days with a total of 4,806 MRSA-free NHP days).

Prevalence, antimicrobial resistance, and enterotoxin gene profiles of Staphylococcus aureus isolated from mobile phones of the food vendors in Phayao province, Thailand

BackgroundMobile phones are widely used and may cause bacterial pathogens to spread among various professionals. Staphylococcus aureus from the mobile phones can contaminate the hands of food vendors and food during the cooking or packaging process. This research aimed to determine the prevalence, enterotoxin genes, and antimicrobial resistance (AMR) profiles of S. aureus contaminating the vendors’ mobile phones.MethodsIn this study, 266 mobile phone samples were randomly collected from food vendors selling food on walking streets (n = 139) and in food centers (n = 127) in Phayao province. All samples were identified as S. aureus by the conventional culture method and confirmed species-specific gene by polymerase chain reaction (PCR). Then, all identified S. aureus isolates were tested for antimicrobial susceptibility by broth microdilution method and for the presence of staphylococcal enterotoxin (SE) genes by PCR.ResultsThe results showed that 12.8% of the mobile phones collected were contaminated with S. aureus. Of 49 S. aureus isolates obtained, 30 (61.2%) were positive for SE genes. The most common SE gene was sea followed by sec, seb, sem, seq, and sel. Moreover, S. aureus was most frequently resistant to penicillin, followed by chloramphenicol and tetracycline, erythromycin, clindamycin, and gentamicin. Methicillin-resistant S. aureus (MRSA), vancomycin-resistant S. aureus (VRSA), and multidrug-resistant (MDR) strains were also detected.ConclusionsThis study showed that mobile phones were an intermediate surface for the transmission of S. aureus, including MDR variants. It indicates that hand hygiene and the decontamination of mobile phones are essential to prevent cross-contamination of S. aureus in food settings.

Staphylococcus aureus Induced Wound Infections Which Antimicrobial Resistance, Methicillin- and Vancomycin-Resistant: Assessment of Emergence and Cross Sectional Study.

Wound infection is a prevalent concern in the medical field, being is a multi-step process involving several biological processes. Methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) infections often occur in areas of damaged skin, such as abrasions and open wounds. This research aims to light the incidence of MRSA and VRSA in wound swabs, the antimicrobial susceptibility configuration of isolated S. aureus patterns in pus/wound samples collected from Saudi Arabian tertiary hospital. The cross section study, β- lactamase detection, VRSA genotyping, MAR index, D-test and VRSA genotyping are methods, which used for completed this research. Patients of several ages and genders delivered specimens from two hospitals in the Al jouf area, in the northern province of Saudi Arabia. S. aureus was found in 188 (34.7%) of the 542 wounds. The traumatized wounds provided 71 isolates (38.8%), surgical wound provided 49 isolates (26.8%) and abscess were represented 16 by isolates (8.7%). In the study, 123 (65.4%) out of 188 were MRSA, 60 (31.9%) were MSSA, and five (2.7%) were VRSA. Linezolid and rifampin were found to be the most effective antimicrobials with 100% in vitro antibacterial activity against S. aureus isolates. The Multiple antimicrobials resistance (MAR) index revealed 73 isolates (38.9%) with a MAR index greater than 0.2, and 115 (61.1%) less than 0.2. The D-test showed that of MLSb phenotypes among S. aureus, 22 (11.7%) strains were D-test positive (MLSbi phenotype), 53 (28.2%) strains were constitutive MLSc phenotypes, and 17 (9%) strains were shown to have MSb phenotypes. All VRSA isolates (n=5) were found to be positive for vanA, and no vanB positive isolates were detected in the study. Regular monitoring and an antimicrobials stewardship program should be in place to provide critical information that can be utilized for empirical therapy and future prevention strategies.

Multidrug resistant and multivirulent avian bacterial pathogens: tackling experimental leg disorders using phytobiotics and antibiotics alone or in combination

Locomotor disorders caused by multidrug-resistant (MDR) bacterial pathogens denote one of the most detrimental issues that collectively threaten the poultry industry leading to pronounced economic losses across the world. Hence, searching for effective alternatives, especially those extracted from plant origins became of great priority targeting a partial or complete replacement of chemical antimicrobials to tackle their developing resistance. Therefore, we aimed to determine the prevalence and antimicrobial resistance of Staphylococcus aureus (S. aureus), Salmonella species, Mycoplasma synoviae (M. synoviae), and Escherichia coli (E. coli) recovered from 500 broilers and ducks (250 each) with locomotor disorders in various farms in Dakahlia and Sharkia Governorates, Egypt. Additionally, we assessed, for the first time, the in vitro antimicrobial effectiveness of marjoram, garlic, ginger and cinnamon essential oils (EOs) against MDR and multivirulent bacterial isolates as well as the in vivo efficiency of the most effective antibiotics and EOs either separately or in combination in the treatment of experimentally induced poultry leg disorders. The overall prevalence rates of S. aureus, E. coli, Salmonella species, and M. synoviae were 54, 48, 36, and 2%, respectively. Salmonella species and S. aureus prevailed among ducks and broilers (36 and 76%, respectively). Notably, MDR was observed in 100, 91.7, 81.1, and 78.5% of M. synoviae, E. coli, Salmonella, and S. aureus isolates, respectively. Our in vitro results displayed that marjoram was the most forceful EO against MDR and multivirulent chicken vancomycin-resistant S. aureus (VRSA) and duck S. Typhimurium isolates. The current in vivo results declared that marjoram in combination with florfenicol or amoxicillin/clavulanic acid succeeded in relieving the induced duck and chicken leg disorders caused by S. Typhimurium and VRSA, respectively. This was evidenced by improvement in the clinical and histopathological pictures with a reduction of bacterial loads in the experimental birds. Our encountered successful in vitro and in vivo synergistic effectiveness of marjoram combined with florfenicol or amoxicillin/clavulanic acid recommends their therapeutic application for leg disorders and offers opportunities for reducing the antibiotics usage in the poultry industry.

Insight into the Antibacterial Action of Iodinated Imine, an Analogue of Rafoxanide: a Comprehensive Study of Its Antistaphylococcal Activity.

In this study, we have focused on a multiparametric microbiological analysis of the antistaphylococcal action of the iodinated imine BH77, designed as an analogue of rafoxanide. Its antibacterial activity against five reference strains and eight clinical isolates of Gram-positive cocci of the genera Staphylococcus and Enterococcus was evaluated. The most clinically significant multidrug-resistant strains, such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant S. aureus (VRSA), and vancomycin-resistant Enterococcus faecium, were also included. The bactericidal and bacteriostatic actions, the dynamics leading to a loss of bacterial viability, antibiofilm activity, BH77 activity in combination with selected conventional antibiotics, the mechanism of action, in vitro cytotoxicity, and in vivo toxicity in an alternative animal model, Galleria mellonella, were analyzed. The antistaphylococcal activity (MIC) ranged from 15.625 to 62.5 μM, and the antienterococcal activity ranged from 62.5 to 125 μM. Its bactericidal action; promising antibiofilm activity; interference with nucleic acid, protein, and peptidoglycan synthesis pathways; and nontoxicity/low toxicity in vitro and in vivo in the Galleria mellonella model were found to be activity attributes of this newly synthesized compound. In conclusion, BH77 could be rightfully minimally considered at least as the structural pattern for future adjuvants for selected antibiotic drugs. IMPORTANCE Antibiotic resistance is among the largest threats to global health, with a potentially serious socioeconomic impact. One of the strategies to deal with the predicted catastrophic future scenarios associated with the rapid emergence of resistant infectious agents lies in the discovery and research of new anti-infectives. In our study, we have introduced a rafoxanide analogue, a newly synthesized and described polyhalogenated 3,5-diiodosalicylaldehyde-based imine, that effectively acts against Gram-positive cocci of the genera Staphylococcus and Enterococcus. The inclusion of an extensive and comprehensive analysis for providing a detailed description of candidate compound-microbe interactions allows the valorization of the beneficial attributes linked to anti-infective action conclusively. In addition, this study can help with making rational decisions about the possible involvement of this molecule in advanced studies or may merit the support of studies focused on related or derived chemical structures to discover more effective new anti-infective drug candidates.

Evidence and Molecular Characterization of Multidrug Resistant Staphylococcus aureus Isolated From Equines in Pakistan

The purpose of the current study was to investigate the nasal colonization and drug resistance profile of Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), and vancomycin-resistant S. aureus (VRSA) in donkeys (n =63), mules (n=42), and horses (n=98). MRSA and VRSA were confirmed based on phenotypic and molecular methods, followed by phylogenetic analysis. Furthermore, the association of various animal and management-based risk factors with S. aureus colonization was also evaluated. The presence of nuc gene on polymerase chain reaction showed an overall prevalence of 42.86% of S. aureus in equines. Based on Kirby-Bauer disc diffusion test, 26.44% of isolates showed resistance to vancomycin. Further, mecA and vanB genes were targeted which showed an overall 14.94% and 13.79% prevalence of methicillin and vancomycin-resistant isolates, respectively. The phylogenetic analysis revealed a significant variation of the study isolates with each other and with already reported sequences of mecA and vanB genes. Risk factor analysis revealed that raising purpose (P=.011), work intensity (P < .001), stocking density (P=.006), presence of other livestock animals in surroundings (P=.043), and common drinking water source (P=.023) as significant. Antimicrobial susceptibility testing of MRSA and VRSA isolates showed high resistance to various commonly used antibiotics. Furthermore, all the tested isolates showing resistance to three or more than three antibiotics were considered multiple drug-resistant. The current study manifests the molecular evidence of MRSA and VRSA isolated from equines in Pakistan which will help to address the emerging issue of multidrug resistance in S. aureus in equines and emphasizes the need for possible measures to tackle this issue.

Shapeshifting bullvalene-linked vancomycin dimers as effective antibiotics against multidrug-resistant gram-positive bacteria

The alarming rise in superbugs that are resistant to drugs of last resort, including vancomycin-resistant enterococci and staphylococci, has become a significant global health hazard. Here, we report the click chemistry synthesis of an unprecedented class of shapeshifting vancomycin dimers (SVDs) that display potent activity against bacteria that are resistant to the parent drug, including the ESKAPE pathogens, vancomycin-resistant Enterococcus (VRE), methicillin-resistant Staphylococcus aureus (MRSA), as well as vancomycin-resistant S. aureus (VRSA). The shapeshifting modality of the dimers is powered by a triazole-linked bullvalene core, exploiting the dynamic covalent rearrangements of the fluxional carbon cage and creating ligands with the capacity to inhibit bacterial cell wall biosynthesis. The new shapeshifting antibiotics are not disadvantaged by the common mechanism of vancomycin resistance resulting from the alteration of the C-terminal dipeptide with the corresponding d-Ala-d-Lac depsipeptide. Further, evidence suggests that the shapeshifting ligands destabilize the complex formed between the flippase MurJ and lipid II, implying the potential for a new mode of action for polyvalent glycopeptides. The SVDs show little propensity for acquired resistance by enterococci, suggesting that this new class of shapeshifting antibiotic will display durable antimicrobial activity not prone to rapidly acquired clinical resistance.

Prevalence and molecular characterization of methicillin resistant Staphylococcus aureus (MRSA) and vancomycin resistant Staphylococcus aureus (VRSA) in a tertiary care hospital

Resistance shown by Staphylococcus aureus to methicillin; mediated by mecA, and vancomycin; mediated by vanA, has led to difficulty in treatment of related infections. Despite reports showing methicillin resistant S. aureus (MRSA) and vancomycin resistant S. aureus (VRSA) in Nepal, and need for their regular surveillance, no study has been conducted on it in our hospital. So, this study is aimed to determine prevalence of MRSA, VRSA and their molecular characterization along with antibiogram. A descriptive cross-sectional study was conducted from August to December, 2022 in Clinical Microbiology Laboratory of NMCTH among S. aureus (n=160) isolated from various clinical specimens after receiving ethical approval from NMC-IRC. AST was done by modified Kirby-Bauer’s disc diffusion method. MRSA and VRSA were detected by cefoxitin disc method and agar dilution method respectively. Inducible clindamycin resistance was detected by D-test. Resistant genes (mecA, PVL, and vanA) were detected using conventional PCR. Prevalence of MRSA was found to be 31.2% (50/160) but none of the isolates were resistant to vancomycin. Total 7 (46.6%) mecA and 7 (46.6%) PVL genes were detected among 15 selected MRSA isolates but vanA was not found. All the MRSA isolates were susceptibile to co-trimoxazole, tigecycline, chloramphenicol, vancomycin, teicoplanin and linezolid. The resistance rate against ciprofloxacin, ofloxacin, and clindamycin was 52.0%, 44.0%, and 68.0% (20.0% iMLSB, 28.0% cMLSB and 16.0% MS-phenotypes) respectively. Prompt implementation of hospital antibiotic policy and AMR Act by government along with regular surveillance of MRSA and VRSA seems essential to contain MRSA infections. Co-trimoxazole could be treatment option against MRSA in our setting, keeping vancomycin in reserve. However, large scale studies are required to establish this conclusion.

Identification of Risk Factors for Nasal Colonization of Methicillin-resistant Staphylococcus aureus and Vancomycin-resistant Staphylococcus aureus in Health Workers at a Tertiary Hospital, Indonesia

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) and Vancomycin-resistant S. aureus (VRSA) frequently cause nosocomial infections yearly. During the COVID-19 pandemic, the potential for excessive use of antibiotics is a global threat to the increasing incidence of multiresistant bacteria. AIM: This study aimed to determine MRSA and VRSA colonization and identify factors associated with the risk of MRSA and VRSA nasal colonization in health workers at Dr. Soemarno Sosroatmodjo General Hospital, Kuala Kapuas, as one of the type C hospitals in Indonesia. METHODS: This cross-sectional analytic study at Dr. Soemarno Sosroatmodjo General Hospital, a tertiary hospital in Indonesia. A 128 health workers’ subjects had undergone nasal swab screening for MRSA and VRSA colonization examinations. Then, they were asked to complete a questionnaire concerning the risk factors of MRSA and VRSA infections. RESULTS: Nasal swab results obtained as many as 30 (23.5%) MRSA positive subjects and 6 (4.7%) subjects with positive VRSA. The most common risk factors that led to MRSA colonization included a history of positive MRSA in the previous hospital (60%), a history of ear, nose, and throat infection (41.7%), and did not do hand rub/handwash (36.7%). In comparison, the most risk factors for VRSA colonization were having pigs farm at home (33.3%), a history of positive MRSA in the previous hospital (20%), and a history of hospitalization in the past 6–12 months (16.7%). The results of multivariate analysis showed the most powerful and statistically significant risk factors in influencing nasal MRSA colonization were a history of positive MRSA in the previous hospital (OR 13.69, 95% confidence intervals [CI]: 1.34–140.25, p = 0.028) and did not do hand rub/handwash (OR 2.95, 95% CI: 1.167–7.49, p = 0.023). Meanwhile, marital status (OR 0.160, 95% CI: 0.02–1.06), p = 0.058) and home care service (OR 6.10, 95% CI: 0.79–46.96, p = 0.082) were the strongest risk factors for nasal colonization of VRSA but not statistically significant. CONCLUSION: As many as, 23.5% and 4.7% of healthcare workers’ subjects were found with nasal colonization of MRSA and VRSA, respectively. Accordingly, strict policies are needed to minimize the transmission of these organisms from the hospital setting to the community.

Molecular Basis of Methicillin and Vancomycin Resistance in Staphylococcus aureus from Cattle, Sheep Carcasses and Slaughterhouse Workers

Staphylococcus aureus (S. aureus) is a serious infection-causing pathogen in humans and animal. In particular, methicillin-resistant S. aureus (MRSA) is considered one of the major life-threatening pathogens due to its rapid resistance to several antibiotics in clinical practice. MRSA strains have recently been isolated in a number of animals utilized in food production processes, and these species are thought to be the important sources of the spread of infection and disease in both humans and animals. The main objective of the current study was to assess the prevalence of drug-resistant S. aureus, particularly vancomycin-resistant S. aureus (VRSA) and MRSA, by molecular methods. To address this issue, a total of three hundred samples (200 meat samples from cattle and sheep carcasses (100 of each), 50 hand swabs, and 50 stool samples from abattoir workers) were obtained from slaughterhouses in Egypt provinces. In total, 19% S. aureus was isolated by standard culture techniques, and the antibiotic resistance was confirmed genotypically by amplification nucA gen. Characteristic resistance genes were identified by PCR with incidence of 31.5%, 19.3%, 8.7%, and 7% for the mecA, VanA, ermA, and tet L genes, respectively, while the aac6-aph gene was not found in any of the isolates. In this study, the virulence genes responsible for S. aureus' resistance to antibiotics had the highest potential for infection or disease transmission to animal carcasses, slaughterhouse workers, and meat products.

Isolation and Detection of Drug-Resistant Bacterial Pathogens in Postoperative Wound Infections at a Tertiary Care Hospital in Saudi Arabia.

Surgical site infections (SSIs), especially when caused by multidrug-resistant (MDR) bacteria, are a major healthcare concern worldwide. For optimal treatment and prevention of antimicrobial resistance, it is important for clinicians to be aware of local drug-resistant bacterial pathogens that cause SSIs. To determine the frequency patterns of drug-resistant bacterial strains causing SSIs at a tertiary care hospital in Saudi Arabia. This retrospective study was conducted at the Microbiology laboratory of Al-Noor Specialist Hospital, Makkah, Saudi Arabia, and included wound swab samples from all cases of SSI between January 01, 2017, and December 31, 2021. The swabs were processed for the identification of bacterial strains and their resistance pattern to antibiotics according to the Clinical and Laboratory Standards Institute. A total of 5409 wound swabs were analyzed, of which 3604 samples (66.6%) were from male. Most samples were from the Department of Surgery (43.3%). A total of 14 bacterial strains were isolated, of which 9 were Gram-negative bacteria. The most common isolates were Klebsiella pneumoniae, followed by Pseudomonas aeruginosa, Escherichia coli, Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococci (VRE), and vancomycin-resistant S. aureus (VRSA). In terms of MDR in 2021, the highest rate of carbapenem-resistance was in A. baumannii (97%). MDR was as follows: A. baumannii, 97%; K. pneumoniae, 81%; E. coli, 71%; MRSA, 60%; P. aeruginosa, 33%; VRE, 22%; and VRSA, 2%. This study showed that in the city of Makkah, Saudi Arabia, the rates of MDR bacteria are high, with the majority being Gram-negative.

Biofilm forming multidrug resistant Staphylococcus aureus of dairy origin: molecular and evolutionary perspectives.

Biofilm production by Staphylococcus aureus is a prevailing cause of multidrug resistance. The evolutionary mechanisms of adaption with host and pathogenicity are poorly understood. The present study aimed to investigate the biofilm-forming potential, associated multidrug resistance, and the evolutionary analysis of S. aureus isolated from bovine subclinical mastitis. 122 S. aureus isolates were subjected to Congo red agar method (CRA), microtitre plate method (MTP), and PCR to check the biofilm-forming potential. The Kirby-Bauer disk diffusion method was used to evaluate the antibiotic resistance pattern. The icaA gene of isolates was subjected to molecular and evolutionary analysis using different bioinformatics tools. The results showed that 63.93% of S. aureus isolates carried the icaA gene and the detection rate of CRA was higher (36.07%) compared to the MTP test (24.59%). A total of 78.21% and 56.41% of biofilm-positive isolates were methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA), respectively. All S. aureus isolates (100%) showed multidrug resistance. The molecular analysis showed an evolutionary link between isolates and revealed a strong codon bias, three different recombination events, and positive selection in some residues of the semi-conserved segments of the icaA gene. The study concluded that biofilm-positive isolates have a high tendency to exhibit methicillin, vancomycin, and multidrug resistance. The findings suggest that mutation and selection are the most likely causes of codon bias in the icaA gene sequences. The variations led by recombination events and positive selection are suggestive of bacterial strategy to combat antimicrobial effects and to escape the host's immune surveillance.

Antibacterial and antibiofilm activities of taxifolin against vancomycin-resistant S. aureus (VRSA)

Background and objective: The medicinal effects of flavonoids are widely described in the literature; however, their antimicrobial effects against antibiotic resistant bacteria are yet to be highlighted. This study was aimed at investigating the growth and biofilm inhibitory effects of taxifolin, a flavonoid, against vancomycin-resistant Staphylococcus aureus (VRSA). Methods: Seven VRSA isolates were used to assess the antimicrobial and antibiofilm influence of taxifolin. The agar-well diffusion method was used to determine the zones of inhibition caused by taxifolin, and resazurin-based microdilution technique was used to assess the minimum inhibitory concentration. Crystal violet staining technique was used to assess the biomass of biofilms formed by the microorganisms. GraphPad Prism software was used to present the data in figures. Results: Taxifolin inhibited bacterial growth in a dose-dependent fashion and reduced bacterial viability. It similarly attenuated the biofilm production activity of bacterial isolates in a dose-dependent manner. Conclusions: Current findings suggest the antibacterial and antibiofilm influence of taxifolin against VRSA in a dose-dependent manner.