Search Results (215)

Introduction: Klebsiella variicola, a member of Klebsiella pneumoniae complex, has emerged as an opportunistic pathogen for human infection; however, antimicrobial resistance and hypervirulent characteristics of K. variicola have rarely been investigated in South Korea. Methods: We analyzed 76 clinical K. variicola isolates collected from 12 hospitals between September 2022 and October 2023. Bacterial identification was performed by MALDI-TOF MS. Antimicrobial susceptibility was tested by disk diffusion tests. Resistance determinants and virulence traits were investigated, and whole-genome sequencing was performed for hypermucoviscous or carbapenem-resistant K. variicola isolates. Results: Most (89.5%, 68/76) were susceptible to all 18 antimicrobials tested in this study, and 3 isolates harbored bla_CTX-M-15. One isolate carried bla_KPC-2 on its IncX3 plasmid, which is closely related to carbapenem-resistant K. pneumoniae plasmids. Capsular typing revealed 51 wzi allelic types. Ten isolates showed mucoid phenotype, mainly with KL60 and KL61. Conclusions: This study reveals relatively low resistance rates in K. variicola strains but the presence of multidrug-resistant and hypermucoviscous K. variicola strains. In addition, the evidence of interspecies dissemination of bla_KPC-2 highlights the need for continuous genomic surveillance. Full article

Clostridioides difficile was first identified in 1935 and subsequently emerged over the next several decades as the predominant bacterial cause of healthcare-associated gastrointestinal infections, placing a significant burden on healthcare systems worldwide. A major driver of the rapid rise in the incidence of C. difficile infection (CDI) was the emergence and spread of a hypervirulent strain, which became known as PCR ribotype 027 (RT027). The C. difficile RT027 strain produced not just the typical toxin A and toxin B virulence factors but also expressed a third toxin called binary toxin that enhanced pathogenicity. Interestingly, the C. difficile RT027 strain apparently emerged at least twice in geographically distinct areas. The two lineages can be differentiated by their resistance or susceptibility to fluoroquinolones. Other ribotypes of C. difficile that also express binary toxin have emerged recently, some of which are highly related genetically to RT027. The aim of this study is to integrate genomic data and published literature to clarify the emergence, divergence, and apparent decline of C. difficile RT027. Full article

Hypervirulent Klebsiella pneumoniae (hvKp) has emerged as a significant public health concern, yet rare sublineages remain poorly characterized. Here, we described a K30-ST198 hvKp sublineage identified in four isolates from two patients, including three sequential strains (K30B1, K30B2, K30B3) recovered over eight months from recurrent liver abscesses and one strain (K30-HUMV1) from a urinary tract infection. All isolates exhibited a yYpermucoviscous phenotype and resistance restricted to ampicillin and amoxicillin. Screening with the eazyplex hvKp assay detected ybt and rmpA in all strains, yielding a virulence score of 1. Biofilm production was strong in K30B1, K30B2, moderate in K30-HUMV1, but weak in K30B3. In the Galleria mellonella infection model, K30B1 showed higher virulence than the other isolates. Whole-genome sequencing identified the ICEKp1 carrying hypervirulence-associated genes (ybt, pagO, rmpAC, iroBCDN) together with additional virulence factors (fim, mrkD, uge, ureA, wabG, wcaJ, mliC), while antibiotic resistance genes were limited to fosA and bla_SHV-77. Protein structures and their functional domains were predicted using AlphaFold v3.0.1 and ColabFold v1.5.5, based on pLDDT scores, providing further insights into gene functionality. This work represents one of the first detailed characterizations of K30-ST198 hvKp, underscoring the need for integrated genomic, phenotypic, and structural approaches in hvKp surveillance. Full article

This study evaluated the diffusion of Listeria monocytogenes (LM) in a beef steak tartare production chain, aiming to (1) evaluate Listeria spp. diffusion in finishing farms supplying beef cattle, (2) evaluate LM prevalence in carcasses, and (3) map LM diffusion in the production plant. A detection rate of 6/76 was observed in the farm, while carcasses after skinning and before refrigeration tested positive in 19/30 and 11/30, respectively. During tartare production, 57/154 meat and 35/191 environmental samples tested positive. A total of 114 LM isolates were characterized via a whole-genome sequencing approach. Five clonal complexes (CCs) and seven sequence types (STs) were identified, with CC9-ST580 being the most prevalent. Four clusters were identified from both the slaughtering and production phases. Genes related to resistance to fosfomycin, quinolones, sulfonamides, lincosamide, and tetracycline were detected. Two hypervirulent strains (CC6-ST6 and CC2-ST145), harboring a full-length inlA, several virulence genes, and stress islands, were detected. Stress Survival Islet 1 was found in almost all the isolates. The wide diffusion of LM in steak tartare requires the management of some critical phases of the production chain (mainly slaughtering); genomic methodologies could be useful in describing the circulation and virulence of LM strains. Full article

Background/Objectives: Streptococcus pyogenes (Group A Streptococcus, GAS) is a major human pathogen that causes a wide spectrum of diseases. While mild infections like pharyngitis and impetigo are common, severe and invasive infections, though less frequent, pose significant health risks, particularly in children. In recent years, the re-emergence of hypervirulent GAS strains has heightened global concern. Nowadays, the absence of universally accepted guidelines compels clinicians to rely on a combination of clinical judgment, microbiological data and available evidence to manage these infections effectively. Our aim was to assess the current management of invasive GAS (iGAS) infections in Italy and the variability in therapeutic and preventive approaches. Methods: A web-based current clinical practice survey about invasive and severe GAS infections was designed according to the Checklist for Reporting of Survey Studies (CROSS) methodology and circulated among the members of the Italian Society of Pediatric Infectious Diseases (SITIP). Results: The survey reveals that while many practices are commonly shared among clinicians, particularly regarding first-line therapies (penicillin or ceftriaxone depending on the infection site), significant uncertainties remain, particularly about the use of combined antibiotic regimens and supportive treatments. The use of combined antibiotic regimens was considered appropriate as first-line therapy for STSS, NF and brain abscesses. Clindamycin was the preferred agent for combination with beta-lactam for most infections, except for brain abscesses, where linezolid was favored. However, there was disagreement regarding the optimal timing for de-escalation to beta-lactam monotherapy. Responses varied widely concerning the indications and dosages for IVIG, as well as the use of corticosteroids. Conclusions: Addressing the burden of invasive GAS (iGAS) infections in children requires enhanced surveillance, early recognition, prompt treatment and preventive strategies. Further work to increase surveillance, e.g., developing national registries, and to standardize the management of the disease, e.g., developing country-specific guidelines, is essential to build solid evidence on the most effective approaches. Full article

Plesiomonas shigelloides, a Gram-negative bacterium prevalent in aquatic environments and also frequently isolated from livestock and poultry, was investigated through integrated whole-genome sequencing and functional analyses. We deciphered the pathogenic mechanisms of P. shigelloides CA-HZ1, a highly virulent strain isolated from a novel piscine host, revealing a complete genome assembly with a 3.49 Mb circular chromosome and 311 kb plasmid housing 3247 predicted protein-encoding genes. Critical genomic features included 496 virulence factors and 225 antibiotic resistance genes. Pathogenicity analysis indicated that P. shigelloides was responsible for disease outbreaks. Antimicrobial susceptibility tests showed resistance to various drugs, such as kanamycin, erythromycin, and penicillin. Histopathological examination showed significant alterations in the infected hosts. Quantitative real-time PCR (qRT-PCR) was carried out to analyze immune-related gene (IL-6, IL-1β, IL-21, STAT1, and HSP70) levels in liver and intestinal tissues, demonstrating the potent immunity triggered by P. shigelloides infection. An analysis of the liver transcriptome revealed that P. shigelloides has the potential to influence the cellular composition, molecular functions, and biological processes. Collectively, this study describes the genomic basis underlying both the pathogenic potential and hypervirulence of P. shigelloides CA-HZ1, establishing a foundational framework for investigating its broad host tropism and immune response. Full article

Background: The emergence and spread of the tigecycline resistance gene tet(A)-v1 in carbapenem-resistant Klebsiella pneumoniae (CRKP) poses significant public health challenges. However, the prevalence of tet(A)-v1-positive CRKP, especially in pediatric patients, remains poorly understood. This study aims to address the gap by performing an in-depth analysis of isolates collected from a children’s hospital in China. Methods: A 4-year retrospective study was conducted in the children’s hospital in Suzhou, China. Non-duplicated specimens were obtained from pediatric patients, and antimicrobial susceptibility profiles were assessed. Whole-genome sequencing and bioinformatics analyses were conducted to characterize the genetic background, antimicrobial resistance determinants, hypervirulence-associated genes, diversity of tet(A)-v1-carrying plasmids, the genetic environment of tet(A)-v1, and the potential for clonal transmission. Conjugative transferability of tet(A)-v1-carrying plasmids was also evaluated via conjugation assays. Results: Of the 73 tet(A)-v1-positive CRKP isolates from pediatric patients, 10.96% were non-susceptible to tigecycline. These isolates exhibited high genetic diversity, spanning across 13 STs (sequence types), with ST17 being predominant. Three carbapenemases were identified, with IMP being the most common. Isolates from diverse backgrounds, such as ST17, ST20, ST323, ST792, and ST3157, demonstrated evidence of clonal transmission. The tet(A)-v1 gene was located on 14 distinct plasmids across seven replicon types, with IncFIA/IncHI1 and IncFII being most commonly detected. All tet(A)-v1-carrying plasmids were multidrug-resistant, and 68.49% were conjugatively transferable, indicating a high potential for horizontal transfer. Four genetic contexts bordering tet(A)-v1 were identified, which points to active clonal dissemination. Conclusions: Although limited to a single hospital, this study represents one of the first in-depth investigations of tet(A)-v1-positive CRKP in pediatric patients, providing valuable insights into the prevalence and spread of tet(A)-v1 in this vulnerable group. These findings emphasize the urgent need for enhanced surveillance and infection control measures to curb the spread of tet(A)-v1-positive CRKP in pediatric healthcare environments, offering critical insights to mitigate its public health impact. Full article

The global emergence of multidrug- and pandrug-resistant Klebsiella pneumoniae poses a critical threat to public health, particularly in hospital settings. This study describes a nosocomial outbreak caused by K. pneumoniae in a tertiary-care hospital in Mexico and provides a comprehensive genomic analysis of six clinical isolates. All isolates exhibited pandrug resistance, including carbapenems and colistin. Whole-genome sequencing identified 37 antimicrobial resistance genes, including blaNDM-1, blaOXA-1, blaCTX-M-15, and a pmrB R256G mutation associated with colistin resistance. Two conjugative plasmids (pAA046 and pAA276) carried multiple resistance genes and mobile genetic elements. Although all isolates harbored CRISPR-Cas type I-E systems, no spacers matched resistance plasmids, suggesting functional inactivity. Capsular typing identified the KL27 locus with the wzi187 allele. Phylogenetic and cgMLST analyses confirmed clonal dissemination and close genetic relatedness to strains from Europe and the USA. Despite the absence of classical hypervirulence markers, the presence of kfu, fimH, and mrkD genes indicates adaptation to the hospital environment. These findings confirm the clonal spread of pandrug-resistant K. pneumoniae ST392-KL27 in a Mexican hospital, underscoring the role of plasmid-mediated resistance and the potential for global dissemination. Full article

Streptococcus pyogenes (Group A Streptococcus, GAS) is a major human pathogen capable of causing infections ranging from mild pharyngitis and impetigo to severe invasive diseases such as bacteremia, necrotizing fasciitis, and streptococcal toxic shock syndrome (STSS). Historically, the incidence of GAS infections declined during the early antibiotic era but began rising again from the early 2000s, driven partly by the emergence of hyper-virulent strains such as emm1 and emm12. From 2005 onward, significant increases in GAS infections were reported globally, accompanied by rising antibiotic resistance, particularly to macrolides and tetracyclines. During the COVID-19 pandemic, widespread public health measures led to a sharp decline in GAS infections, including invasive cases, but this trend reversed dramatically in late 2022 and 2023, with surges exceeding pre-pandemic levels, notably in children. Recent data implicate factors such as “immunity debt,” viral co-infections, and the spread of virulent clones like M1UK. Looking forward, continued surveillance of GAS epidemiology, virulence factors, and resistance patterns is critical. Moreover, the emergence of GAS isolates with reduced susceptibility to beta-lactams underscores the need for vigilance despite the absence of fully resistant strains. The development of an effective vaccine remains an urgent priority to reduce GAS disease burden and prevent severe outcomes. Future research should focus on vaccine development, molecular mechanisms of virulence, and strategies to curb antimicrobial resistance. Full article

Antimicrobial resistance (AMR), particularly due to extended-spectrum β-lactamases (ESBLs) and carbapenemases (CPs), poses a critical threat to global health. This study aimed to characterize the molecular epidemiology, resistance profiles, and genomic features of ESBL- and CP-producing Escherichia coli and Klebsiella pneumonaie (ESBL/CP-Ec/Kp) isolates from a Spanish hospital (2020–2024) and explore links to environmental reservoirs like white storks foraging at a nearby landfill. A total of 121 clinical Ec/Kp isolates (55 ESBL-Ec, 1 CP-Ec, 35 ESBL-Kp, 17 CP-Kp, 13 ESBL+CP-Kp) underwent phenotypic testing, PCR, and whole-genome sequencing (WGS). Analyses included phylogenomics (cgMLST), detection of AMR genes, plasmid typing, and comparative genomics. Among ESBL-Ec, bla_CTX-M-15 was the most prevalent (60.0%), and one CP-Ec carrying bla_NDM-5 was identified. WGS of 44 selected ESBL/CP-Ec isolates revealed a variety of AMR genes, and 56.8% of isolates carried class one integrons (56.8%). IncF-type plasmids predominated, and 84.1% of isolates were assigned as ExPEC/UPEC. The lineage ST131 dominated (75%), with IncF-bla_CTX-M-15-carrying plasmids. Among the 18 ESBL/CP-Kp isolates sequenced, the lineage ST307 was the most frequent (44.4%), followed by ST15 and ST11, carrying a diversity of AMR determinants and plasmids (IncFIB(K), IncL, ColpVC). Virulence included ybt loci in ICEKp; hypervirulence genes were absent. Genomic analysis of 62 clinical isolates (44 Ec, 18 Kp) showed close phylogenetic links to stork-derived strains, with ST131-Ec and ST307-Kp from humans and birds differing just by ≤22 and ≤10 ADs, respectively, with a conserved plasmid content (i.e., IncL-bla_OXA-48, IncFIB(K)-bla_CTX-M-15). High-risk ESBL/CP-Ec/Kp clones persist across clinical and environmental contexts. WGS-based surveillance is key for understanding AMR spread and guiding interventions. Results support a One Health approach to combat AMR through cross-sector collaboration. Full article

Carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKp), a pathogen causing severe nosocomial infections and high mortality rates, is increasingly becoming a serious global public health threat. Capsular polysaccharide (CPS), a major virulence factor of hvKp, can be enzymatically degraded by bacteriophage-derived depolymerases. However, to our knowledge, depolymerases targeting K. pneumoniae K54-type strains have rarely been identified. Here, we identified and characterized three novel capsule depolymerases, Dep_C, Dep_Y, and Dep_Z, derived from three different K. pneumoniae phages, which retained robust activity across a broad pH range (pH 3.0–12.0) and demonstrated thermal stability up to 50 °C. These depolymerases could efficiently digest the CPS of K. pneumoniae K54-serotype strains, significantly inhibit biofilm formation, and remove their mature biofilms. Although no bactericidal activity was detected, these depolymerases rendered host bacteria susceptible to serum complement-mediated killing. We further demonstrate that Dep_C, Dep_Y, and Dep_Z can effectively and significantly prolong the survival time of mice in a pneumonia model infected with K54-type K. pneumoniae and reduce the colonization and virulence of the bacteria in the mice. These findings indicate that depolymerases Dep_C, Dep_Y, and Dep_Z could increase bacterial susceptibility to host immune responses of hvKp to the host through their degradation effect on the CPS. In conclusion, our study demonstrates that the three capsule depolymerases are promising antivirulent agents to combat CR-hvKp infections. 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

Aeromonas hydrophila ST251 is a crucial pathogen responsible for the outbreaks of Motile Aeromonas Septicemia (MAS) in global aquaculture. To elucidate the genetic basis underlying its hypervirulence, we investigated strain D4, an ST251 isolate recovered from diseased blunt snout bream. Phenotypic assays revealed that, compared to the environmental strain ATCC 7966^T, D4 exhibited enhanced motility, hemolytic activity, and protease production. Average nucleotide identity (ANI) analysis demonstrated that D4 clustered within a distinct ST251 clade, with ANI values ≥ 99.74%. Comparative genomic analysis of D4, nine additional ST251 strains, and ATCC 7966^T identified multiple unique genomic islands in ST251 strains, including pathways for myo-inositol and L-fucose utilization and a pseudaminic acid biosynthesis gene cluster. These genetic elements are associated with nutrient acquisition and flagellar assembly, potentially enhancing colonization and environmental adaptability. In addition, distinct plasmids and prophages in ST251 strains may contribute to host adaptation and virulence by regulating stress responses and virulence-associated genes. These findings offer new insights into the molecular mechanisms driving the pathogenicity and adaptability of hypervirulent A. hydrophila ST251 strains. Full article

The emergence of antimicrobial resistance (AMR) in Clostridium difficile (C. difficile), particularly to last-line antibiotics such as linezolid, represents a critical challenge in clinical settings. This study investigates the genomic epidemiology of linezolid-resistant C. difficile, focusing on the distribution and mutational patterns of the chloramphenicol–florfenicol resistance (cfr) gene and its association with multidrug resistance. We analyzed 514 clinical isolates (354 from NCBI Pathogen Detection, 160 from EnteroBase), revealing distinct prevalence patterns among cfr subtypes: cfr(C) was dominant (156/354 NCBI strains; 101/160 EnteroBase strains), whereas cfr(B) frequently harbored missense mutations (p.R247K, p.V294I, and less commonly p.A334T). The cfr(E) subtype was exclusively identified in ribotype 027 (RT027) strains. Notably, cfr(C) exhibited a strong association with RT017, correlating with a conserved 99 bp genomic deletion. Phylogenetic analysis linked cfr-carriage to predominant sequence types (ST1 in NCBI strains, ST37 in EnteroBase isolates). Furthermore, the co-occurrence of cfr with additional AMR genes conferred resistance to macrolides (erythromycin, azithromycin) and tetracyclines, indicating a convergent evolution toward multidrug resistance. These findings underscore the interplay between cfr mutations, hypervirulent ribotypes, and AMR dissemination, necessitating enhanced surveillance to mitigate the spread of resistant C. difficile lineages. Full article

Acinetobacter baumannii has become a formidable pathogen in healthcare systems worldwide, primarily due to its remarkable capacity to develop multidrug resistance and cause life-threatening infections. While traditionally A. baumannii is considered an opportunistic pathogen of low virulence, accumulating evidence now underscores the emergence of hypervirulent A. baumannii (hvAB) strains. These strains combine heightened pathogenicity with extensive drug resistance, posing unprecedented challenges for clinical management and infection containment. This review comprehensively explores the molecular mechanisms driving hvAB’s virulence and antimicrobial resistance and its evolutionary trajectory, clinical presentations, and global epidemiology. Additionally, we evaluate potential therapeutic strategies and their broader public health implications. Full article