Search Results (581)

Introduction: In recent years, antimicrobial resistance (AMR) rates have increased significantly in bacterial pathogens, particularly extended beta-lactam resistance. This study aimed to investigate resistome and phylogenomics of Escherichia coli (E. coli) strains isolated from various sources in Jimma, Ethiopia. Methods: Phenotypic antibiotic resistance patterns of E. coli isolates were determined using automated Kirby–Bauer disc diffusion and minimum inhibitory concentration (MIC). Isolates exhibiting phenotypic resistance to beta-lactam antibiotics were further analyzed with a DNA microarray to confirm the presence of resistance-encoding genes. Additionally, multilocus sequence typing (MLST) of seven housekeeping genes was conducted using PCR and Oxford Nanopore-Technology (ONT) to assess the phylogenetic relationships among the E. coli isolates. Results: A total of 611 E. coli isolates from human, animal, and environmental sources were analyzed. Of these, 41.6% (254) showed phenotypic resistance to at least one of the tested beta-lactams, 96.1% (244) thereof were confirmed genotypically. More than half of the isolates (53.3%) had two or more resistance genes present. The most frequent ESBL-encoding gene was CTX-M-15 (74.2%; 181), followed by TEM (59.4%; 145) and CTX-M-9 (4.1%; 10). The predominant carbapenemase gene was NDM-1, detected in 80% (12 out of 15) of carbapenem-resistant isolates. A phylogenetic analysis revealed clonality among the strains obtained from various sources, with international high-risk clones such as ST131, ST648, ST38, ST73, and ST405 identified across various niches. Conclusions: The high prevalence of CTX-M-15 and NDM-1 in multidrug-resistant E. coli isolates indicates the growing threat of AMR in Ethiopia. The discovery of these high-risk clones in various niches shows possible routes of transmission and highlights the necessity of a One Health approach to intervention and surveillance. Strengthening antimicrobial stewardship, infection prevention, and control measures are crucial to mitigate the spread of these resistant strains. Full article

Avian pathogenic Escherichia coli (APEC) poses a global threat to poultry health and public safety due to its high lethality, limited treatment options, and potential for zoonotic transmission via the food chain. However, long-term genomic surveillance remains limited, especially in countries like China where poultry farming is highly intensive. This study aimed to characterize the population structure, virulence traits, and antimicrobial resistance of 81 APEC isolates from diseased chickens collected over 16 years from Shandong and neighboring provinces in eastern China. The isolates were grouped into seven Clermont phylogroups, with A and B1 being dominant. MLST revealed 27 STs, and serotyping identified 29 O and 16 H antigens, showing high genetic diversity. The minor phylogroups (B2, C, D, E, G) encoded more virulence genes and had higher virulence-plasmid ColV carriage, with enrichment for iron-uptake, protectins, and extraintestinal toxins. In contrast, the dominant phylogroups A and B1 primarily carried adhesin and enterotoxin genes. Antimicrobial resistance was widespread: 76.5% of isolates were multidrug-resistant. The minor phylogroups exhibited higher tetracycline resistance (mediated by tet(A)), whereas the major phylogroups showed increased resistance to third- and fourth-generation cephalosporins (due to bla_CTX-M-type ESBL genes). These findings offer crucial data for APEC prevention and control, safeguarding the poultry industry and public health. Full article

Background: Hospitalized COVID-19 patients are particularly vulnerable to secondary bacterial infections, which can significantly worsen clinical outcomes. The aim of the study was to identify the cause of bacteremia in a group of hospitalized COVID-19 patients and find out the source of the outbreak to prevent further spread. Methods: Pathogen identification in blood cultures and sensitivity testing were carried out using the automated VITEK2 system. A total of 110 samples were tested; these were collected from patients’ colonization sites and from surfaces, materials and fluids used in the setting. Furthermore, multilocus sequence typing (MLST) and next-generation sequencing (NGS) were employed to characterize the isolates. Results: Achromobacter xylosoxidans was detected in the blood of nine hospitalized patients and in cotton used for disinfection; all isolates presented an identical antibiotic resistance pattern, and all carried the bla_OXA-114 gene which is intrinsic to this species. Infection control measures were implemented promptly. With one exception, all patients recovered and were discharged in good health. Conclusions: This outbreak underscores the urgent need for investigation and control of hospital infections, as bacteremia is associated with increased morbidity, mortality, hospitalization time, and cost. It also highlights the importance of close collaboration among healthcare professionals. Full article

Klebsiella pneumoniae, a zoonotic pathogen of global concern, poses significant threats to both veterinary and public health. Here, a comparative study characterized 14 clinical isolates (7 avian-derived, 7 human-derived) from Jiangsu, China, through integrated genomic and phenotypic analyses. Firstly, multilocus sequence typing (MLST) revealed distinct epidemiological patterns: the same ST type in avian isolates was circulating between different species and different regions, whereas it was not found in human isolates. In addition, hypervirulent Klebsiella pneumoniae (hvKP) phenotypes confirmed by string test were exclusive to two human isolates (KP15, KP20). Secondly, biofilm detection demonstrated 78.6% (11/14) of isolates possessed biofilm-forming capacity, with cellulose but not curli as the predominant matrix component. Human-derived KP15 and KP20 had the strongest biofilm formation ability in all isolates. Antimicrobial susceptibility profiling identified serious multidrug resistance in both avian and human isolates. Virulence gene analysis revealed striking disparities, with human isolates harboring 10–20 virulence factors (median 15) versus 6–7 (median 6.5) in avian counterparts. Finally, functional pathogenesis assessments demonstrated human-derived strains exhibited stronger epithelial cell adhesion (2-fold higher) and invasion (1.97-fold higher) in Calu-3 cell models and paradoxically showed reduced macrophage phagocytosis (2.85-fold lower at 2 h) for immune escape. In vivo models confirmed dose-dependent mortality, with human isolates demonstrating higher lethality in both Galleria mellonella and mice. Virulence gene burden positively correlated with mortality outcomes. These findings delineate critical host adaptation differences in Klebsiella pneumoniae populations and provide empirical evidence for pathogen transmission dynamics at the human-animal interface. Full article

The aim of this study was to examine the genetic characteristics that could be associated with the virulence characteristics of Escherichia coli collected from clinical samples. A collection of 100 non-repetitive E. coli isolates was analyzed. All isolates were typed by MLST. String production, biofilm formation and serum resistance were examined for all isolates. Twenty E. coli isolates were completely sequenced Illumina platform. The results showed that the majority of E. coli isolates (87%) produced significant levels of biofilm, while none of the isolates were positive for string test and resistance to serum. Additionally, the presence of CRISPR/Cas systems (type I-E or I-F) was found in 18% of the isolates. Analysis of WGS data found that all sequenced isolates harbored a variety of virulence genes that could be implicated in adherence, invasion, iron uptake. Also, WGS data confirmed the presence of a wide variety of resistance genes, including ESBL- and carbapenemase-encoding genes. In conclusion, an important percentage (87%) of the E. coli isolates had a significant ability to form biofilm. Biofilms, due to their heterogeneous nature and ability to make microorganisms tolerant to multiple antimicrobials, complicate treatment strategies. Thus, in combination with the presence of multidrug resistance, expression of virulence factors could challenge antimicrobial therapy of infections caused by such bacteria. Full article

Staphylococcus lugdunensis is a coagulase-negative staphylococcus known for its significant pathogenic potential, often causing severe infections such as endocarditis and bacteremia, with virulence comparable to S. aureus. Despite general susceptibility to most antibiotics, the emergence of oxacillin-resistant strains is increasingly concerning. This study conducted whole-genome sequencing on 20 S. lugdunensis isolates from Chang Gung Memorial Hospital to explore their genetic diversity, antimicrobial resistance mechanisms, and mobile genetic elements. The lugdunin biosynthetic operon, essential for antimicrobial peptide production, was present in multilocus sequence typing (MLST) types 1, 3, and 6 but absent in STs 4, 27, and 29. Additionally, IS256 insertion elements, ranging from 7 to 17 copies, were identified in four strains and linked to multidrug resistance. CRISPR-Cas systems varied across STs, with type III-A predominant in ST1 and ST6 and type IIC in ST4, ST27, and ST29; notably, ST3 lacked CRISPR systems, correlating with a higher diversity of SCCmec elements and an increased potential for horizontal gene transfer. Phage analysis revealed stable phage–host associations in ST1, ST6, and ST29, whereas ST4 displayed a varied prophage profile. Phenotypic resistance profiles generally aligned with genomic predictions, although discrepancies were observed for aminoglycosides and clindamycin. These findings highlight the complex genetic landscape and evolutionary dynamics of S. lugdunensis, emphasizing the need for genomic surveillance to inform clinical management and prevent the spread of resistant strains. Full article

Campylobacter jejuni is a leading cause of bacterial gastroenteritis worldwide which usually presents as mild, and self-limiting disease in immunocompetent individuals. However, in immunocompromised patients, such as those with common variable immunodeficiency, C. jejuni can cause severe recurrent infections requiring antibiotic treatment. Our study reports a case of a 37-year-old male patient with CVID, who had multiple episodes of C. jejuni intestinal infections over a 3.5-year period. A total of 27 stool samples were collected and analyzed between December 2020 and July 2024 during acute febrile diarrheal episodes, with C. jejuni isolated in 15 samples. Antimicrobial susceptibility testing (AST) during the course of the disease revealed three different antimicrobial resistance profiles including multi-drug-resistant phenotype. Whole genome sequencing was performed on three representative isolates, all identified as MLST type 367, ST-257 complex, with minimal genetic divergence, indicating a clonal origin. Genes and point mutations conferring resistance to macrolides, fluoroquinolones, beta-lactams, and tetracycline were identified in different C. jejuni isolates, along with key virulence factors linked to adherence, invasion, motility, and immune evasion. The genetic analysis of macrolide phenotypic resistance revealed different resistance mechanisms. Genotypic and phenotypic analyses of the same C. jejuni clone from single patient, and identified multidrug resistance pattern, present the first documented case of in vivo resistance development of C. jejuni in Croatia. This case highlights the role of prolonged antibiotic pressure in driving resistance evolution and underscores the need for careful antimicrobial stewardship and genomic monitoring in immunocompromised patients. Further research is needed to correlate phenotypic resistance with genetic determinants in Campylobacter spp. Full article

Staphylococcus aureus is an important cause of food intoxication, which has the potential to induce diverse infections, toxinoses and life-threatening diseases among humans and animals. This study investigated the prevalence, antimicrobial resistance, and genetic diversity of S. aureus and methicillin-resistant S. aureus (MRSA) in retail raw meat from Shandong (March 2021–October 2022). The distribution of virulence genes, antimicrobial susceptibility, and genetic diversity of these isolates were analyzed. From a total of 442 samples, 87 (19.7%) S. aureus and 11 (2.5%) MRSA were isolated. According to the antimicrobial susceptibility testing, it was found that all the S. aureus isolates were resistant to at least one antimicrobial. Most isolates (95.9%) were resistant to penicillin, with high resistance to ampicillin (82.7%) and multidrug resistance in 76.5% of cases. One isolate could simultaneously resist eleven antimicrobials (ERY-CLI-GEN-SMZ-FFC-PEN-PRL-AMC-CIP-TET-AMP). In contrast, all the isolates showed sensitivity to vancomycin. The most prevalent virulence gene was sed, accounting for 10.2%, followed by sec (8.2%). Regarding genetic polymorphism, these isolates were divided into 21 different sequence types (STs) using multilocus sequence typing (MLST) and 33 staphylococcal protein A (spa) types using spaTyper 1.0 tool. The most prevalent sequence types were ST398 (22.4%), followed by ST7 (20.4%), while ST59, ST1, ST188, ST9, ST398, and ST7 were observed in MRSA isolates. The most prevalent spa types were t034 (15.3%), followed by t899 (10.2%), while t441, t127, t184, t899, t034, and t091 were observed in MRSA isolates. In conclusion, our study highlights the high prevalence of S. aureus and MRSA in different retail raw meats in Shandong. This poses a potential threat to food safety and underscores the need for enhanced surveillance and stricter antibiotic control measures. Full article

Shiga-toxin-producing E. coli (STEC) is a leading causing of bacterial foodborne and zoonotic illnesses in the USA. Whole-genome sequencing (WGS) is a powerful tool used in public health and microbiology for the detection, surveillance, and outbreak investigation of STEC. In this study, we applied three WGS-based subtyping methods, high quality single-nucleotide polymorphism (hqSNP) analysis, whole genome multi-locus sequence typing using chromosome-associated loci [wgMLST (chrom)], and core genome multi-locus sequence typing (cgMLST), to isolate sequences from 11 STEC outbreaks. For each outbreak, we evaluated the concordance between subtyping methods using pairwise genomic differences (number of SNPs or alleles), linear regression models, and tanglegrams. Pairwise genomic differences were highly concordant between methods for all but one outbreak, which was associated with international travel. The slopes of the regressions for hqSNP vs. allele differences were 0.432 (cgMLST) and 0.966 wgMLST (chrom); the slope was 1.914 for cgMLST vs. wgMLST (chrom) differences. Tanglegrams comprised of outbreak and sporadic sequences showed moderate clustering concordance between methods, where Baker’s Gamma Indices (BGIs) ranged between 0.35 and 0.99 and Cophenetic Correlation Coefficients (CCCs) were ≥0.88 across all outbreaks. The K-means analysis using the Silhouette method showed the clear separation of outbreak groups with average silhouette widths ≥0.87 across all methods. This study validates the use of cgMLST for the national surveillance of STEC illness clusters using the PulseNet 2.0 system and demonstrates that hqSNP or wgMLST can be used for further resolution. Full article

Background: During the COVID-19 pandemic, the emergence of multidrug-resistant (MDR) pathogens, driven by heightened antibiotic usage and device-associated infections, has posed significant challenges to healthcare. This study reports an outbreak of Proteus mirabilis producing NDM-5 and CTX-M-15 β-lactamases in a hospital in Buenos Aires, Argentina, from October 2020 to April 2021. To our knowledge, this represents the first documented outbreak of NDM-5-producing P. mirabilis in the country. Methods: A total of 82 isolates were recovered from 40 patients, with 41.5% from blood cultures and 18.3% from respiratory and urinary samples, among others. Antimicrobial susceptibility testing, PCR-based methods, and MALDI-TOF MS cluster analysis were conducted. Whole genome sequencing (WGS) was performed to characterize the MLST, resistome and plasmid content. Biofilm formation assays and in vitro rifampicin susceptibility tests were also conducted. Result: Most isolates exhibited resistance to carbapenems, cephalosporins, aminoglycosides, and fluoroquinolones, while retaining susceptibility to aztreonam. Genetic analysis confirmed the co-presence of the bla_NDM-5 and bla_CTX-M-15 genes. Clonal relationships was supported by PCR-based typing and MALDI-TOF MS cluster analysis. WGS revealed a resistome comprising 25 resistance genes, including rmtB and both β-lactamases, as well as the presence of an incomplete IncQ1 replicon associated with multiple resistance determinants. MLST classified this clone as belonging to ST135. Despite the biofilm-forming capacity observed across strains, rifampicin demonstrated potential for disrupting established biofilms at concentrations ≥32 µg/mL in vitro. The MDR profile of the outbreak strain significantly limited therapeutic options. Conclusions: This study highlights the growing threat of NDM-producing P. mirabilis in Argentina. The absence of surveillance cultures from the index case limits insights into the outbreak’s origin. These findings underscore the importance of integrating genomic surveillance into infection control protocols to mitigate the spread of MDR pathogens. Full article

Background/Objectives: Pseudomonas aeruginosa is one of the most significant multidrug-resistant bacteria. It poses considerable challenges in terms of treatment and causes hospital-acquired infections that lead to high morbidity and mortality. Colonization by P. aeruginosa in a patient without clinical signs of infection is a concern in hospital settings, as it is an opportunistic pathogen and can potentially be a multidrug-resistant strain. The objective of this study was to characterize and provide a detailed genomic analysis of this strain of the P. aeruginosa PSU9449 genome, an isolate obtained from a patient at Songklanagarind Hospital, Thailand. Methods: Whole-genome sequencing (WGS) and bioinformatics analysis were employed to examine the genomic features of P. aeruginosa PSU9449. We performed sequence type (ST) determination through multilocus sequence typing (MLST), identified antimicrobial resistance genes (ARGs), virulence factor genes (VFGs), and analyzed the presence of mobile genetic elements (MGEs). Additionally, we compared the PSU9449 genome with strains from neighboring countries to understand its phylogenetic relationship. Results: The P. aeruginosa PSU9449 genome contained five insertion sequences and several ARGs, including fosA, aph (3’)-IIb, bla_OXA-50, and catB7. It also harbored VFGs related to flagella (fli, fle, and flg), the type 6 secretion system (hcpA, tssA, and las), and the type 3 secretion system (exoS, exoU, and exoT). MLST identified PSU9449 as ST3777, which was reported in Thailand for the first time. Phylogenetic analysis based on core gene SNPs revealed that PSU9449 was closely related to P. aeruginosa HW001G from Malaysia and P. aeruginosa MyJU45 from Myanmar, forming a distinct clade. Conclusions: This study presents a comprehensive genomic analysis of P. aeruginosa PSU9449, shedding light on its genetic characteristics, antimicrobial resistance profile, and virulence potential. Interestingly, ST3777, the novel STs from the published genomes of P. aeruginosa in Thailand, were assigned in this study. The findings enhance valuable insights into the expanding knowledge of P. aeruginosa PSU9449 and highlight the importance of ongoing surveillance of its genetic diversity. Full article

The global emergence of carbapenem-resistant Acinetobacter baumanii (CRAB) represents a significant public health threat. In the summer of 2022, a polyclonal CRAB outbreak occurred in our hospital, marking the first detection of an NDM-1 plus OXA-23 co-producing A. baumannii strain in Spain. The aim of this study was to phenotypically and genotypically characterize the clonal spread of NDM-1 and OXA-23 co-producing A. baumannii isolates and to describe the infection control measures implemented to contain the outbreak. Patients with multidrug-resistant A. baumannii isolates (July 2022–May 2023) were included in the study. Isolates were identified via MALDI-TOF, and antimicrobial susceptibility was tested using a broth microdilution method (DKMGN SensititreTM panels). Whole-genome sequencing was performed on 24 representative isolates. Phylogenetic analysis was performed using Ridom SeqSphere+ (cgMLST), while sequence typing was performed using ARIBA (Pasteur and Oxford schemes). A. baumannii isolates from the affected patients belonged to five different sequence types. The two main STs were ST1Pas/ST231Oxf (NDM-1- and OXA-23-co-producing), which accounted for 58%, and ST136Pas/ST406Oxf (OXA-23-producing), which accounted for 21%. All isolates were resistant to fluoroquinolones, trimethoprim/sulfamethoxazole, aminoglycosides, and carbapenems. In addition, 8% were resistant to colistin and 17% to cefiderocol. Finally, the affected patients were cohorted, and a thorough cleaning of the affected units was carried out. This study documents the clonal spread of an NDM-1- and OXA-23-co-producing A. baumannii strain in Spain, linked to a Libyan patient, highlighting the risk of cross-border spread. Although infection control measures successfully contained the outbreak, surveillance is essential as the incidence of CRAB infections is expected to increase. Full article

This study was undertaken to determine the effect of bird age and administering either Lactococcus lactis ssp. lactis 1 (LL) or Limosilactobacillus fermentum 1 (LF) in the drinking water on the prevalence of antimicrobial resistance by phenotypic test, multilocus sequence typing (MLST) and virulence genes of Escherichia coli (E. coli) isolated from broiler caeca by whole-genome sequencing (WGS) analysis. Male (Ross 308) day-old chicks (240) were reared for 28 days. Water was provided either untreated (CON) or with LL (10⁷/mL) or LF (10⁷/mL) via a nipple drinker on three days each week during the starter phase (days 1, 3, 5, 7, 9 and 11 d) in eight replicate pens per treatment, with initially ten chicks per pen. One chick from each pen was sacrificed when LL or LF was added to the water, and again on d 14 and 28. There was no evidence that LL and LF had any effect on the prevalence of antimicrobial resistance and virulence genes in E. coli isolates. The population density of Lactobacillus sp. and coliforms decreased with age (p < 0.001). The high resistance of E. coli to ampicillin and tetracycline was maintained throughout the life of the broilers. The prevalence of virulence genes was greatest during the starter phase but declined when birds were 28 days of age (p < 0.05). In birds < 14 d of age, E. coli MLST 457, 1640, 1485 and 155 were dominant, and these carried iucD, irp2, astA, iutA and iroN genes. When birds were 28 d of age, MLST 1286, 1112 and 973 predominated, and these carried few virulence genes. This suggests that young birds were more susceptible to putative pathogenic E. coli than older birds. Supporting the development of a healthy microbiome that might control the proliferation of potentially pathogenic E. coli is an area of future research. Full article

Nakaseomyces glabratus is a medically important fungal pathogen responsible for various opportunistic, life-threatening, and fatal infections, mainly among immunodepressed patients worldwide. Herein, genotypes identified in Central Poland by multilocus sequence typing (MLST) are presented. Along with the genotyping, drug susceptibility was performed. The research was conducted on 30 non-redundant clinical strains, and 15 distinct sequence types (STs) were identified, including three novel STs: ST212, ST213, and ST214. The most prevalent sequence types were ST3, ST6, and ST10. Antifungal susceptibility testing revealed varied resistance rates to azoles, with fluconazole susceptibility at 16.7% and high susceptibility to amphotericin B. No correlation between ST and antifungals MIC were found. The study findings highlight the genetic diversity of N. glabratus in Central Poland and the role of surveillance and research to elucidate antifungals resistance and molecular epidemiology of N. glabratus. Full article

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of pediatric infections and has shown evolving molecular characteristics over time. This study aimed to investigate the phenotypic and genotypic features of MRSA isolates collected from pediatric patients at a tertiary medical center in northern Taiwan between 2011 and 2020. A total of 182 MRSA strains were analyzed for SCCmec types, PVL gene presence, antimicrobial susceptibility, multilocus sequence typing (MLST), and clonal relatedness using pulsed-field gel electrophoresis (PFGE). ST59/SCCmec Vt was the most prevalent genotype, followed by ST59/SCCmec IV and ST8/SCCmec IV. Most ST59/SCCmec Vt and ST8/SCCmec IV isolates clustered genetically. Clindamycin and erythromycin resistance remained high, whereas co-trimoxazole susceptibility ranged from 76% to 100%. These findings confirm ST59 as the dominant clone and highlight the emergence of ST8 and ST45 in community-associated MRSA (CA-MRSA) infections. Oral co-trimoxazole remains the most effective empirical option, while clindamycin and erythromycin should be avoided. Continuous molecular surveillance is warranted to monitor trends and guide treatment strategies in pediatric MRSA infections. Full article