Multidrug Resistance Profiles Research Articles

Draft genome sequence of a co-harbouring blaNDM-5 and mcr-1.1 Escherichia coli phylogroup A isolate associated with patient colonisation in Ireland.

ObjectivesWhile Escherichia coli phylogroup-A is typically associated with commensal strains, some isolates can harbour virulence and exhibit multidrug-resistant (MDR) phenotypes. We report the draft genome of a rare instance of carbapenem, fosfomycin and colistin resistant E. coli phylogroup-A, isolated as part of routine screening of a human patient in a clinical setting in Ireland. MethodsE. coli E230738 was identified using MALDI-ToF/MS. Antibiotic susceptibility testing was performed using the Sensitire-EUMDRXXF plate. Whole-genome-sequencing was conducted with NextSeq1000, and genomic analysis identified antibiotic-resistance-genes (ARGs) and virulence-factors (VFs). Phylogenetic analysis was performed using whole-genome-multilocus-sequence-typing (wgMLST). ResultsE. coli E230738 genome was identified to belong to phylogroup-A/ST10 complex and to harbour 63 ARGs (17 acquired). Resistance to beta-lactams, including carbapenems and cephalosporins was likely due to predicted chromosomal blaNDM-5. Colistin resistance appeared associated with acquired mcr-1.1. Despite lacking fosfomycin-inactivating-enzymes, fosfomycin resistance was observed, possibly due to efflux pumps. 47 chromosomal VFs were identified, involved in adhesion and iron acquisition amongst others. Plasmid replicons associated with the spread of MDR genes such as IncHI2/HI2A were detected. Phylogenetic analysis showed the closest relative being a strain from the UK differing by 851 genes. ConclusionThis is a first detected instance of a blaNDM-5 and mcr-1.1 co-occurring in E. coli in Ireland. The MDR profile of E. coli E230738 highlights the growing public health threat posed by the dissemination of MDR E. coli lineages with limited treatment options and underscores the need for clinical screening coupled with genomic surveillance to better understand evolving MDR patterns in E. coli.

Prevalence, Cross Contamination, Virulence, and Multidrug Resistance Profiles of Staphylococcus aureus Isolates from Four Middle-Scale Dairy Farms in Bareilly, Northern India.

Milk, a nutritious global important food commodity, serves as an excellent medium for microbial growth as well. The foodborne pathogen Staphylococcus aureus is a commensal member of human microflora that enters the food chain through poor hygienic practices and cross contamination and causes various clinical manifestations in humans. During this study, raw milk and swab samples (milker's hand, udder, towel, milking bucket, and farm floor) were collected from four middle-scale buffalo dairy farms. The results revealed S. aureus presence in 11.6% (n = 56/448) bucket raw milk samples and 2.6% (n = 12/448) udder raw milk samples. Contrarily, S. aureus prevalence was significantly higher in farm floors (100%, n = 84/84), towel (35.7%, n = 10/28), milking bucket (11.6%, n = 56/448), milker's hand (10.7%, n = 3/28), and udder swab samples (4.0%, n = 18/448). The chi-square test yielded p values of 0.000, 0.005, and 0.0011 for udder raw milk, udder swab, and milker's hand swab, respectively. The p values of the milking bucket (p = 0.048) and farm floors (p = 0.0183) confirmed their possible role in S. aureus cross contamination. Gene amplifications of nuclease and enterotoxin A indicate potential virulence of S. aureus isolates in collected samples. Antibiotic susceptibility testing revealed multidrug resistance in 44% (n = 239) of S. aureus isolates with the highest resistance of 61% against penicillin. Resistance against ampicillin, streptomycin, and lincomycin was observed. Fewer S. aureus isolates were resistant to kanamycin and erythromycin, whereas the lowest number of resistant isolates was observed against chloramphenicol. A high prevalence of S. aureus in the farm environment and milking equipment suggested the cross contamination of potentially enterotoxin-producing and multidrug-resistant S. aureus to raw milk. Therefore, good hygiene practices should be enforced to avoid foodborne and zoonotic infections.

Assessing the Occurrence of Carbapenemase Producers Using Marine Animals as Sentinel Species

Carbapenemase-producing (CP) strains represent a substantial global threat, deactivating carbapenems and conferring resistance to β-lactam antibiotics. They can spread across various environments, yet data on their presence in marine animals are sparse. This study aimed to assess the occurrence of carbapenemase-producing strains in wild marine animals and to analyse their antimicrobial resistance (AMR) profiles to crucial antimicrobials and heavy metals frequently encountered in the marine environment due to anthropogenicactivity. A total of 28 samples were obtained from a fish auction in the Centre Region of Portugal. Non-fermenting bacilli (NFB) was isolated from the visceral content of wild marine animals. Identification of isolates was achieved through PCR-based amplification of the 16S rRNA gene, followed by sequencing. Antimicrobial susceptibility testing was conducted according to EUCAST guidelines, covering nine antimicrobials. Research of carbapenemases and metal tolerance genes was conducted by PCR, and statistical analysis utilized the Fisher’s exact test. Pseudomonas spp. and Aeromonas spp., among other isolates were identified (n=47/9/7, respectively). Susceptibility profiles showed 100% resistance or intermediate resistance to ticarcillin, piperacillin, piperacillin-tazobactam, ceftazidime, ciprofloxacin, and imipenem (n=63/63), while 27% were resistant to meropenem (n=17/63) and 13% to tobramycin (n=8/63). All of them exhibited susceptibility to amikacin and carried multidrug resistance (MDR) profiles, including heavy metal genes (merA and silA). None harboured the carbapenemase genes searched (blaKPC, blaGES, blaIMP, blaNDM or/and blaVIM). In this study, MDR profiles to clinically important antimicrobials were observed, including to carbapenems. However, no carbapenemase-producing strains were identified, suggesting the presence of other genes or alternative mechanisms of resistance. These findings underscore the importance of monitoring AMR in marine ecosystems, particularly given its close ties to the food chain.

Overview of Antimicrobial Resistant ESKAPEE Pathogens in Food Sources and Their Implications from a One Health Perspective.

Antimicrobial resistance is an increasing societal burden worldwide, with ESKAPEE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species and Escherichia coli) pathogens overwhelming the healthcare sectors and more recently becoming predominantly a concern for their persistence in food and food industries, including agricultural settings and animal husbandry environments. The aim of this review is to explore the mechanisms by which the ESKAPEE group gained its multidrug resistance profiles, to analyse their occurrence in different foods and other related reservoirs, including water, and to address the current challenges due to their spread within the food production chain. Moreover, the repertoire of surveillance programmes available focused on monitoring their occurrence, common reservoirs and the spread of antimicrobial resistance are described in this review paper. Evidence from the literature suggests that restricting our scope in relation to multidrug resistance in ESKAPEE pathogens to healthcare and healthcare-associated facilities might actually impede unveiling the actual issues these pathogens can exhibit, for example, in food and food-related reservoirs. Furthermore, this review addresses the need for increasing public campaigns aimed at addressing this challenge, which must be considered in our fight against antimicrobial resistance shown by the ESKAPEE group in food and food-related sectors.

Stenotrophomonas maltophilia bacteremia in hemato-oncological patients

Abstract Background Stenotrophomonas maltophilia is an increasingly common cause of bacteremia in immunocompromised patients, particularly those with underlying neutropenia. This study aims to characterize the clinical presentation, risk factors, and outcomes of Stenotrophomas maltophilia bacteremia in pediatric hemato-oncological patients. Methods We retrospectively reviewed the medical records of nine pediatric hemato-oncological patients who developed a cluster of Stenotrophomas maltophilia bacteremia at our institution from September to November in 2023. The identification and antibiotic susceptibility testing was performed by Vitek 2. Data collected included patient demographics, clinical presentation, underlying disease, treatment regimens, and outcomes. Results We identified nine cases of Stenotrophomas maltophilia bacteremia during the study period. The median age of the patients was 11 years (range 3-16 years), 7 were male. All patients had underlying hematologic malignancies (particularly acute lymphoblastic leukemia) and had profound neutropenia (ANC <100/mm3) at the time of bacteremia. The most common clinical presentation were fever (100%) and the most frequent risk factors were having a central venous catheter (CVC) (100%) and early chemotherapy treatment prior the diagnosis, median of 9 days (range 3-14 days). All patients received intravenous antibiotics, and the most commonly used agent was trimethoprim-sulfamethoxazole (TMP-SMX), the median duration of treatment was 12 days (range 7-16 days). The overall mortality rate was 11.1% on a patient with necrotizing fasciitis. Conclusion Stenotrophomas maltophilia bacteremia is a severe complication in pediatric hemato-oncological patients, considering its multidrug-resistance profile. The rise in Stenotrophomonas maltophilia isolations has triggered the implementation of outbreak investigation measures and exhibits the importance of an adequate control in central line care. Early diagnosis and appropriate antimicrobial therapy are crucial for improving outcomes.

Antimicrobial resistance in Escherichia coli and Staphylococcus aureus at human-animal interfaces on Chongming Island, Shanghai: A One Health perspective

Antimicrobial resistance (AMR) is a significant concern within the One Health framework due to its ability to spread across multiple interfaces. Phenotypic data remains the primary type for AMR surveillance, but exploring association across multiple interfaces poses certain challenges. In this study, AMR phenotypic data of clinical and food animal E. coli and S. aureus from Chongming Island over the past five years were analyzed to determine key characteristics of AMR and explore its association at the human-animal interface.The clinical E. coli isolates showed significant resistance to penicillins (83.92 %), cephems (63.05 %), fluoroquinolones (62.21 %), and tetracyclines (57.77 %), while S. aureus exhibited high resistance to penicillinase-labile penicillins (90.89 %), macrolides (51.51 %), penicillinase-stable penicillins (43.96 %), and lincosamides (43.55 %). Extended-spectrum β-lactamase (ESBL)-producing E. coli isolates accounted for 53.26 % (1398/2526), while methicillin-resistant Staphylococcus aureus (MRSA) prevalence was 43.81 % (435/993). Notably, there has been an increase in the proportion of E. coli isolates resistant to 8 to 12 antimicrobial classes, and in the proportion of S. aureus isolates resistant to 5 to 9 classes. Certain multi-drug resistance (MDR) phenotypes were first identified in food animal isolates and later emerged in clinical settings. Meanwhile, several MDR phenotypes were shared between the two interfaces, with 44 identified in E. coli and 12 in S. aureus. Further co-occurrence analysis in E. coli and S. aureus identified several co-occurrence phenotypic pairs or clusters, potentially mediated by a single plasmid or multiple plasmids within a bacterium, indicating potential associations at the human-animal interface.To summarize, a heightened prevalence of MDR in clinical E. coli and S. aureus has been observed, with some MDR profiles appearing in food animals before emerging in clinical settings. The co-occurrence of phenotypic pairs or clusters underscores the potential for AMR association and transmission between humans and food animals. Within the One Health framework, integrating genomic data into AMR monitoring is a crucial next step.

Determination of Pathogenic E. coli and Antimicrobial Resistance Genes in dogs and human Semen: Evidence of Multidrug-Resistance and Antimicrobial Resistance Gene Profiles

Determination of Pathogenic E. coli and Antimicrobial Resistance Genes in dogs and human Semen: Evidence of Multidrug-Resistance and Antimicrobial Resistance Gene Profiles

Antimicrobial Susceptibility of Enterococci Isolated from Nestlings of Wild Birds Feeding in Supplementary Feeding Stations: The Case of the Canarian Egyptian Vulture.

Antimicrobial resistance is a growing concern worldwide, requiring a holistic "One Health" strategy to address the interconnectedness of human, animal, and environmental health. This study focused on Enterococci isolated from Canary Island Egyptian vulture chicks, an endangered species that feeds at supplementary feeding stations in the Canary Islands. Sampling and identification revealed the presence of several Enterococcus species, with a predominance of E. faecalis. Antimicrobial susceptibility testing showed resistance patterns, especially to important antibiotics such as quinolones, vancomycin, and linezolid. The prevalence of multidrug-resistant profiles was lower than that in other wild bird species. This study underscores the need for further research to understand the dynamics of antimicrobial resistance in wildlife and its implications for public health and conservation efforts, emphasizing the importance of a "One Health" approach to address this pressing problem.

Identification and phylogenetic analysis of carbapenemase genes from clinical strains of Klebsiella pneumoniae.

Klebsiella pneumoniae is an encapsulated Gram-negative bacterium that is responsible for numerous infections in healthcare facilities worldwide and is frequently isolated. The World Health Organization has listed K. pneumoniaeas as a critical antibiotic resistant bacterial pathogen for which new antibiotics are urgently needed. This study aimed to use molecular tools to identify and examine antibiotic resistance in clinical strains of K. pneumoniae. A total of 15 unduplicated K. pneumoniae strains isolated from patient samples with multidrug-resistant (MDR) profiles were subjected to polymerase chain reaction (PCR) to amplify the most common carbapenem resistance genes. (GTG)5 PCR and phylogenetic analysis were performed to identify the genetic relationship between the strains. All strains yielded a (GTG)5-PCR profile, and this allowed us to group these strains into 8 groups according to the size and number of characteristic bands. Phylogenetic analysis was done using the free software UPGMA and a single bacterial clone with a correlation coefficient of over 97% was identified. New Delhi metallo-beta-lactamase NDM-like (blaNDM) carbapenem resistance genes were detected in three strains of K. pneumoniae, which represented a resistance rate of 20%. However, carbapenemases type A [Klebsiella pneumoniae carbapenemase (KPC) and imipenem-hydrolysing beta-lactamase (IMI), type D [oxacillinase-48 (OXA-48)], and other metallo-β-lactamase [Verona integron-encoded metallo-beta-lactamase (VIM), and enzyme active on imipenem (IMP)] were not detected. We identified and grouped the blaNDM resistance genes of Klebsiella pneumonia strains.

Antimicrobial, Antioxidant and Anti-Inflammatory Activities of the Mucus of the Tropical Sea Slug Elysia crispata.

Elysia crispata (Sacoglossa, Gastropoda) is a tropical sea slug known for its ability to incorporate functional chloroplasts from a variety of green macroalgae, a phenomenon termed kleptoplasty. This sea slug, amenable to laboratory cultivation, produces mucus, a viscous secretion that serves diverse purposes including protection, locomotion, and reproduction. In this study, we profiled the antimicrobial, antioxidant, and anti-inflammatory activities of the mucus of this sea slug. Results revealed inhibitory activity against several bacterial strains, more pronounced for Gram-negative bacteria. Particularly interesting was the strong inhibitory effect against Pseudomonas aeruginosa, a bacterial species classified by the WHO as a high-priority pathogen and associated with high-risk infections due to its frequent multidrug-resistant profile. Similar inhibitory effects were observed for the mucus native protein extracts, indicating that proteins present in the mucus contributed significantly to the antimicrobial activity. The mucus also showed both antioxidant and anti-inflammatory activities. The latter activities were associated with the low molecular weight (<10 kDa) fraction of the mucus rather than the native protein extracts. This study opens the way to further research on the biotechnological applications of the mucus secreted by this unique marine organism, particularly as an antimicrobial agent.

Virulence Genes Prevalence and Enterobacterial Repetitive Intergenic Consensus‐PCR Profiles of Goose‐Derived Campylobacter jejuni Isolates

ABSTRACTCampylobacter jejuni is a causative agent of gastroenteritis in humans worldwide, and wild and domestic poultry carry of this bacterium in their gastrointestinal tract. Molecular studies to determine the pathogenicity, origin, and epidemiological relationships among C. jejuni isolates from poultry such as chicken, turkey, and goose consumed as human food are important for public health and infection control. This study aimed to investigate the prevalence of virulence genes and Enterobacterial Repetitive Intergenic Consensus (ERIC‐PCR) based genotyping of C. jejuni isolates obtained from goose cloacal swab samples. For this purpose, PCR analysis of flaA, racR, dnaJ, pldA, cadF, cdtC, ciaB, cdtB, cdtA, virB11, and wlaN virulence genes and ERIC‐PCR analysis of 50 C. jejuni isolates were performed. The emerged genetic profiles and antimicrobial resistance genes regarding the isolates were interpreted with the existing multi‐drug resistance (MDR) findings. Virulence gene positivity was detected as 88%, 84%, 82%, 82%, 80%, 80%, 72%, 30%, and 18% for flaA, racR, dnaJ, pldA, cadF, cdtC, ciaB, cdtB, and cdtA, respectively. VirB11 and wlaN genes were not detected among the C. jejuni isolates. Virulence genes‐based genotyping revealed that the C. jejuni isolates exhibited 22 profiles (A–V). As a result of ERIC‐PCR analysis, the C. jejuni isolates showed heterogeneous distribution, exhibiting 14 different ERIC‐PCR profiles (Cluster I [Cl‐I]–Cluster XIV [Cl‐XIV]). The MDR positivity was detected in 7 (14%) of the C. jejuni isolates. Tetracycline and ciprofloxacin were the antibiotics most frequently included in the MDR profiles. There was no clear correlation between ERIC‐PCR profiles, virulence gene profiles, and MDR profiles. However, isolates with triple‐MDR resistant to ampicillin, tetracycline, and ciprofloxacin showed significant heterogeneity in both ERIC‐PCR profile and virulence gene‐based genetic profile, all of which were positive for ciaB and flaA genes. These results indicate that carriage of the C. jejuni isolates with high gene prevalence and MDR profiles by geese may pose a risk for Campylobacter infections in humans.

Epibiotic Bacteria Isolated from the Non-Indigenous Species Codium fragile ssp. fragile: Identification, Characterization, and Biotechnological Potential.

Due to their richness in organic substances and nutrients, seaweed (macroalgae) harbor a large number of epiphytic bacteria on their surfaces. These bacteria interact with their host in multiple complex ways, in particular, by producing chemical compounds. The released metabolites may have biological properties beneficial for applications in both industry and medicine. In this study, we assess the diversity of culturable bacterial community of the invasive alga Codium fragile ssp. fragile with the aim to identify key groups within this epiphytic community. Seaweed samples were collected from the Northern Tunisian coast. A total of fifty bacteria were isolated in pure culture. These bacterial strains were identified by amplification of the ribosomal intergenic transcribed spacer between the 16S and the 23S rRNA genes (ITS-PCR) and by 16S rRNA sequencing. Antimicrobial activity, biochemical, and antibiotic resistance profile characterization were determined for the isolates. Isolated strains were tested for their antimicrobial potential against human and fish bacterial pathogens and the yeast Candida albicans, using the in vitro drop method. About 37% of isolated strains possess antibacterial activity with a variable antimicrobial spectrum. Ba1 (closely related to Pseudoalteromonas spiralis), Ba12 (closely related to Enterococcus faecium), and Bw4 (closely related to Pseudoalteromonas sp.) exhibited strong antimicrobial activity against E. coli. The isolated strain Ba4, closely related to Serratia marcescens, demonstrated the most potent activity against pathogens. The susceptibility of these strains to 12 commonly used antibiotics was investigated. Majority of the isolates were resistant to oxacillin, cefoxitin, tobramycin, and nitrofurantoin. Ba7 and Ba10, closely related to the Vibrio anguillarum strains, had the highest multidrug resistance profiles. The enzymes most commonly produced by the isolated strains were amylase, lecithinase, and agarase. Moreover, nine isolates produced disintegration zones around their colonies on agar plates with agarolitic index, ranging from 0.60 to 2.38. This investigation highlighted that Codium fragile ssp. fragile possesses an important diversity of epiphytic bacteria on its surface that could be cultivated in high biomass and may be considered for biotechnological application and as sources of antimicrobial drugs.

In vitro efficacy of lavender oil, otological gel and gentamicin to eradicate biofilm produced by Pseudomonas aeruginosa.

Otitis externa (OE) is one of the most frequently diagnosed dermatological diseases in dogs, having a multifactorial aetiology. Among the bacterial agents associated with canine OE, Pseudomonas aeruginosa is of special concern owing to its frequent multidrug resistance profile and ability to form biofilms related to the infection's chronicity and recurrence. The main objective of this study was to evaluate and compare the antibiofilm activity of two innovative antimicrobials-an otological gel containing a synthetic antimicrobial peptide and Lavandula angustifolia essential oil-with gentamicin (a conventional antibiotic) using biofilm-producing P. aeruginosa isolates obtained from dogs with OE. The biofilm eradication capacity of gentamicin, otological gel and lavender oil was determined against a collection of 12 P. aeruginosa biofilm-producers among 35 clinical isolates obtained from the ear canals of dogs with OE. Also, the antimicrobial activity of the otological gel against P. aeruginosa biofilms was assessed in an invitro model of dog cerumen. Lavender oil showed the best effectiveness after 30 min of contact, eradicating 58.3% (seven of 12) of the isolates, and gentamicin showed full eradication (12 of 12) after 24 h. The otological gel acted more slowly than the lavender oil; yet at 24 h, the antibiofilm capacity of both compounds was similar, with no significant difference between them. It also was found that triglycerides from synthetic cerumen earwax had antipseudomonal activity and, when combined with the otological gel, led to the full eradication of P. aeruginosa. The results of this invitro study indicate that lavender oil and the otological gel may be effective topical treatments for canine OE promoted by P. aeruginosa biofilm-producers, as alternatives to gentamicin.

Insight into Virulence and Mechanisms of Amphotericin B Resistance in the Candida haemulonii Complex.

The Candida haemulonii complex includes emerging opportunistic human fungal pathogens with documented multidrug-resistance profiles. It comprises Candida haemulonii sensu stricto, Candida haemulonii var. vulnera, Candida duobushaemulonii, Candida pseudohaemulonii, and Candida vulturna. In recent years, rates of clinical isolation of strains from this complex have increased in multiple countries, including China, Malaysia, and Brazil. Biofilm formation, hydrolytic enzymes, surface interaction properties, phenotype switching and cell aggregation abilities, extracellular vesicles production, stress response, and immune evasion help these fungi to infect the host and exert pathological effects. Multidrug resistance profiles also enhance the threat they pose; they exhibit low susceptibility to echinocandins and azoles and an intrinsic resistance to amphotericin B (AMB), the first fungal-specific antibiotic. AMB is commonly employed in antifungal treatments, and it acts via several known mechanisms. Given the propensity of clinical Candida species to initiate bloodstream infections, clarifying how C. haemulonii resists AMB is of critical clinical importance. This review outlines our present understanding of the C. haemulonii complex's virulence factors, the mechanisms of action of AMB, and the mechanisms underlying AMB resistance.

Retrospective ANalysis of multi-drug resistant Gram-nEgative bacteRia on veno-venous extracorporeal membrane oxygenation. The multicenter RANGER STUDY

BackgroundVeno-venous extracorporeal membrane oxygenation (V-V ECMO) is a rapidly expanding life-support technique worldwide. The most common indications are severe hypoxemia and/or hypercapnia, unresponsive to conventional treatments, primarily in cases of acute respiratory distress syndrome. Concerning potential contraindications, there is no mention of microbiological history, especially related to multi-drug resistant (MDR) bacteria isolated before V-V ECMO placement. Our study aims to investigate: (i) the prevalence and incidence of MDR Gram-negative (GN) bacteria in a cohort of V-V ECMOs; (ii) the risk of 1-year mortality, especially in the case of predetected MDR GN bacteria; and (iii) the impact of annual hospital V-V ECMO volume on the probability of acquiring MDR GN bacteria.MethodsAll consecutive adults admitted to the Intensive Care Units of 5 Italian university-affiliated hospitals and requiring V-V ECMO were screened. Exclusion criteria were age < 18 years, pregnancy, veno-arterial or mixed ECMO-configuration, incomplete records, survival < 24 h after V-V ECMO. A standard protocol of microbiological surveillance was applied and MDR profiles were identified using in vitro susceptibility tests. Cox-proportional hazards models were applied for investigating mortality.ResultsTwo hundred and seventy-nine V-V ECMO patients (72% male) were enrolled. The overall MDR GN bacteria percentage was 50%: 21% (n.59) detected before and 29% (n.80) after V-V ECMO placement. The overall 1-year mortality was 42%, with a higher risk observed in predetected patients (aHR 2.14 [1.33–3.47], p value 0.002), while not in ‘V-V ECMO-acquired MDR GN bacteria’ group (aHR 1.51 [0.94–2.42], p value 0.090), as compared to ‘non-MDR GN bacteria’ group (reference). Same findings were found considering only infections. A larger annual hospital V-V ECMO volume was associated with a lower probability of acquiring MDR GN bacteria during V-V ECMO course (aOR 0.91 [0.86–0.97], p value 0.002).Conclusions21% of MDR GN bacteria were detected before; while 29% after V-V ECMO connection. A history of MDR GN bacteria, isolated before V-V ECMO, was an independent risk factor for mortality. The annual hospital V-V ECMO volume affected the probability of acquiring MDR GN bacteria.Trial Registration ClinicalTrial.gov Registration Number NCTNCT06199141, date 12.26.2023.

Proven anti-virulence therapies in combating methicillin- and vancomycin-resistant Staphylococcus aureus infections.

Despite years of efforts to develop new antibiotics for eradicating multidrug-resistant (MDR) and multi-virulent Methicillin-Resistant Staphylococcus aureus (MRSA) and Vancomycin-Resistant Staphylococcus aureus (VRSA) infections, treatment failures and poor prognoses in most cases have been common. Therefore, there is an urgent need for new therapeutic approaches targeting virulence arrays. Our aim is to discover new anti-virulence therapies targeting MRSA and VRSA virulence arrays. We employed phenotypic, molecular docking, and genetic studies to screen for anti-virulence activities among selected promising compounds: Coumarin, Simvastatin, and Ibuprofen. We found that nearly all detected MRSA and VRSA strains exhibited MDR and multi-virulent profiles. The molecular docking results aligned with the phenotypic and genetic assessments of virulence production. Biofilm and hemolysin productions were inhibited, and all virulence genes were downregulated upon treatment with sub-minimum inhibitory concentration (sub-MIC) of these promising compounds. Ibuprofen was the most active compound, exhibiting the highest inhibition and downregulation of virulence gene products. Moreover, in vivo and histopathological studies confirmed these results. Interestingly, we observed a significant decrease in wound area and improvements in re-epithelialization and tissue organization in the Ibuprofen and antimicrobial treated group compared with the group treated with antimicrobial alone. These findings support the idea that a combination of Ibuprofen and antimicrobial drugs may offer a promising new therapy for MRSA and VRSA infections. We hope that our findings can be implemented in clinical practice to assist physicians in making the most suitable treatment decisions.

Characterization of susceptibility patterns and adaptability of the newly emerged Candida auris.

The emergence of Candida auris has caused a major concern in the public health worldwide. This novel fungus is characterized by its multidrug resistance profile, ability to thrive in harsh and stressful conditions, as well as high temperatures and salt concentrations, persistence on hospital surfaces, causing nosocomial infections and outbreaks, and unique fitness properties. Here, we study the antifungal susceptibility patterns, thermotolerance, and halotolerance of 15 putative C. auris clinical isolates from Inkosi Albert Academic Hospital, Durban, South Africa. Five of the C. auris isolates showed resistance to all three antifungals (fluconazole, amphotericin B, and micafungin) and were selected for characterization of their adaptability mechanisms. Four of the tested multidrug-resistant C. auris isolates (C. auris strain F25, C. auris strain F276, C. auris F283, and C. auris M153) showed good growth when exposed to high temperature (42°C) and salinity (10% NaCl) conditions whereas one isolate (C. auris F65) showed moderate growth under these conditions. Candida parapsilosis showed poor growth whereas C. albicans no growth under these conditions. The five C. auris strains were positive for all the adaptive features.

Research note: characteristics of blaNDM and mcr-1 co-producing Escherichia coli from retail chicken meat

Carbapenems and colistin are vital antimicrobials used to treat Enterobacteriaceae-caused infections. The present study aimed to characterize the coexistence mechanism of carbapenem and colistin resistance in an Escherichia coli isolated from retail chicken meat. A total of 4 E. coli isolates co-harboring carbapenem resistance gene blaNDM (2 E. coli isolates with blaNDM-5 and 2 with blaNDM-9) and colistin resistance gene mcr-1. Antimicrobial susceptibility testing exhibited that all the 4 E. coli strains had multidrug resistance profile and consistent with the resistance genes they carried. MLST showed that 3 E. coli isolates belonged to a pathogenic E. coli lineage ST354, which is closely associated with human infections and pose a serious threat to public health. Whole genome sequencing (WGS) showed that 4 mcr-1-positive plasmids with sizes of 60.4 kb-67.4 kb all belonged to the IncI2 type. A total of 5 blaNDM-harboring plasmids ranged from 99.0 kb∼138.3 kb, among which 4 plasmids belonged to unknow type and only pCS5L-NDM belonged to IncFIA/IncFIB group of hybrid plasmids, a novel carrier for blaNDM. Comparative analysis exhibited that the mcr-1 or blaNDM-carrying plasmids of E. coli strains from chicken meat showed high identity with that from Enterobacteriaceae of human origin, which indicated the risk of mcr-1 or blaNDM dissemination from retail meat to human. The simultaneous occurrence of mcr-1 and blaNDM in E. coli emphasizes the significant of antimicrobial resistance surveillance in retail meat.

A lateral flow immunoassay for the rapid identification of Candida auris from isolates or directly from surveillance enrichment broths.

Candida auris is a recently discovered yeast with a multi-drug resistant profile associated with high mortality rates. The rapid identification of Candida auris in hospital settings is crucial to allow appropriate therapeutic and rapid implementation of infection management measures. The aim of this study was to develop a lateral flow immunoassay (LFIA) for the rapid identification of Candida auris. Highly specific monoclonal antibodies were obtained by immunizing mice with membrane proteins from Candida auris which were then used to develop a LFIA whose performance was assessed by testing 12 strains of Candida auris and 37 strains of other Candida species. Isolates were grown on either Sabouraud dextrose, CHROMagarTM Candida Plus or HardyCHROMTM Candida + auris agar plates. The strains were also cultured on salt sabouraud-dextrose with chloramphenicol or a commercially available Salt-Sabouraud Dulcitol Broth with chloramphenicol and gentamicin, and processed using a simple centrifugation protocol to recover a pellet. Finally, the colonies or yeast extract were transferred to the LFIA to determine the specificity and sensitivity of the assay. The LFIA reached 100% specificity and sensitivity from solid agar plates. For both enrichment broths, some Candida non-auris species were able to grow, but the LFIA remained 100% specific. The use of a dextrose-based sabouraud broth resulted in earlier identification with the LFIA, with most of the Candida auris strains detected at 24 h. The developed LFIA prototype represents a powerful tool to fight the emerging threat of Candida auris. Clinical validation represents the next step.

Multi drug resistance profile and antibiofilm activity of garlic oil on Staphylococcus aureus isolated from milk of cattle suffering with mastitis

Multi drug resistance profile and antibiofilm activity of garlic oil on Staphylococcus aureus isolated from milk of cattle suffering with mastitis