Search Results (134)

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

Listeria monocytogenes (L. monocytogenes) is an important foodborne pathogen that poses great risks to food safety and public health, and knowledge about its presence and diversity in potential sources is crucial for effectively tracking and controlling it in the food chain. In this study, we investigated the prevalence, antimicrobial susceptibility, and genomic characteristics of Listeria monocytogenes (L. monocytogenes) collected from retail chicken meat samples in Qingdao, China, in 2022. A total of 38 (10.6%, 38/360) L. monocytogenes isolates were recovered from 360 retail chickens. All 38 isolates were classified into two lineages (I and II), three serogroups (IIa, IIb, IIc), eight sequence types (STs), eight clonal complexes (CCs), eight Sublineages (SLs) and nine cgMLSTs (CTs). ST121 and ST9 were the most prevalent STs in this study. The ST121 strains from China had heterogeneity with those from other countries, while the Chinese ST9 strains had homogeneity with those from other countries. One resistance cassette tet(M)-entS-msr(D) was identified in eight L2-SL121-ST121-CT13265 isolates, the genetic structure of which was identical to that of three reference genomes. All isolates carried the L. monocytogenes pathogenic island (LIPI)-1, with only one carrying LIPI-3 and three carrying LIPI-4. In addition, 11 isolates subtyped as L2-SL121-ST121-CT13265 were found to have a premature stop codon (PMSC) in the inlA gene in this study. Our data revealed the antimicrobial susceptibility, genomic characteristics and evolutionary relationships of L. monocytogenes in retail chicken in Qingdao, China. The characterization of genotypes, virulence, stress and antimicrobial markers of strains circulating in retail chicken in Qingdao, as described in this study, provides the opportunity to improve risk assessments of L. monocytogenes exposure. Full article

Staphylococcus aureus is an opportunistic pathogen, a member of the ESKAPE group, associated with nosocomial infections and foodborne illnesses due to its production of various toxins. This study conducted a comprehensive genomic characterization of S. aureus isolates producing toxic shock syndrome toxin (TSST-1) using a comparative genomics and bioinformatics approach. A total of 166, including 3 bovine mastitis isolates and 163 public genomes, were analyzed. Twenty-eight distinct sequence types (STs) were identified, with ST30 and ST5 being the most prevalent, corresponding to the clonal complexes CC30 and CC5, respectively. Phylogenetic reconstruction revealed two major clades aligned with these complexes, each exhibiting unique virulence factor profiles. Notably, TSST-1 was detected in bovine mastitis genomes, alongside a broad repertoire of virulence markers, such as enterotoxins and secretion system components, posing a potential risk to public health. Additionally, genes related to environmental information processing systems, including ABC transporters and phosphotransferase systems, were prevalent. These results underscore the need for strengthened genomic surveillance and the implementation of both preventive and corrective measures in dairy herds to mitigate zoonotic transmission and ensure food safety. Full article

Purpose: The adenoid microbiota plays a key role in adenoid hypertrophy (AH). This study explored the molecular epidemiology and antimicrobial resistance of Haemophilus. Influenzae (H. influenzae) strains in pediatric AH patients. Methods: Retrospective analysis of pediatric AH patients undergoing endoscopic adenoidectomy. Adenoid tissue samples were cultured to screen for pathogens. H. influenzae strains were identified by 16S rRNA sequencing and serotyped via q-PCR. Multilocus sequence typing (MLST) and ftsI gene analysis were conducted using PubMLST. β-lactamase genes (bla_TEM-1, bla_ROB-1) were detected by PCR, and antibiotic susceptibility testing (AST) was performed using the Etest method. For imipenem-resistant strains, the acrRAB efflux pump gene cluster and ompP2 porin gene were sequenced and compared with those of the wild-type strain Rd KW20. Results: Over 8 months, 56 non-duplicate H. influenzae strains were isolated from 386 patients. The detection rate was highest in children under 5 years (30.5%) compared to those aged 5–10 years (13.4%) and 10–15 years (8.7%). Of 49 sub-cultured strains, all were non-typeable H. influenzae (NTHi). MLST identified 22 sequence types (STs) and 13 clonal complexes (CCs), with CC11 (26.5%), CC3 (14.3%), and CC107 (14.3%) being predominant. Common STs included ST103 (22.4%), ST57 (10.2%), and ST107 (10.2%). Most strains belonged to the ftsI group III-like+ (57.1%). β-lactamase positivity was 98.0% (48/49), with bla_TEM-1 (95.9%) and bla_ROB-1 (18.4%) detected. AST showed low susceptibility to ampicillin (10.2%), amoxicillin–clavulanate (34.7%), azithromycin (12.2%), and trimethoprim–sulfamethoxazole (14.3%). Among the β-lactamase-positive strains, 44/48 were β-lactamase-positive ampicillin-resistant (BLPAR); none were β-lactamase-negative ampicillin-resistant (BLNAR). Imipenem susceptibility was 91.8% (45/49). No carbapenemases were found in the imipenem-resistant strains, but mutations in acrRAB (88.12–94.94% identity) and ompP2 (77.10–82.94% identity) were observed. Conclusions: BLPAR NTHi strains of CC11 are major epidemic strains in pediatric AH. Imipenem resistance in H. influenzae likely results from porin mutations rather than carbapenemase activity. Enhanced surveillance of H. influenzae’s role in AH and its resistance patterns is warranted. Full article

Water buffalo (Bubalus bubalis) are a primary source of milk in Pakistan, where bovine mastitis is a significant health issue among cattle, leading to substantial economic losses. Staphylococcus aureus is a predominant pathogen associated with mastitis; however, a detailed molecular characterization of the strains in the country remains limited. We previously characterized mastitis strains from the Hazara division of Khyber Pakhtunkhwa, Pakistan. In this study, we investigated mastitis cases in the Peshawar division, including samples from both animals and human farm workers for comparison. Higher rates of mastitis (67.27% of animals) and sub-clinical mastitis (91.03% of positive animals) were identified in Peshawar than for those (34.55% and 75.31%, respectively) previously observed in Hazara. Methicillin-susceptible S. aureus (MSSA) belonging to clonal complex 9 (ST2454) were predominant. Methicillin-resistant S. aureus (MRSA) belonging to ST22 and ST8 were also detected in the Nowshera district. While no S. aureus colonization was observed among animal handlers, evidence of hand contamination suggests a potential route for pathogen spread. Low levels of antibiotic resistance were noted amongst isolates, but higher rates were seen in MRSA. This study presents only the second comprehensive molecular investigation of S. aureus isolated from buffalo mastitis in Pakistan and indicates a concerning rise in mastitis within the province. Full article

(1) Background: CRISPR-Cas systems provide adaptive immunity against mobile genetic elements (MGEs) carrying antimicrobial resistance (AMR) genes. Carbapenem-resistant (CR) Klebsiella pneumoniae is a major public health concern, and the role of CRISPR-Cas in its resistance is understudied. This study explored CRISPR-Cas associations with multidrug resistance in clinical K. pneumoniae. (2) Methods: 400 K. pneumoniae isolates (200 CR and 200 carbapenem susceptible (CS)) were analyzed. Carbapenemase genes (bla_OXA-48, bla_NDM-1, bla_KPC-2), cas1, rpoB, and CRISPR1-3 loci were identified by PCR, while only CRISPR loci were sequenced. Genetic relatedness was assessed via PFGE, MLST, and spacer analysis. Statistical analysis utilized chi-squared and Fisher’s exact tests. (3) Results: CRISPR-Cas was present in 15.8% of isolates, mainly subtypes I-E and I-E* (93.3%), with CRISPR3 loci showing the greatest spacer diversity. Clonal complexes ST14/15/101 (CR) and ST35 (CS) were identified. bla_OXA-48 was linked to CRISPR-Cas-negative strains, while bla_NDM-1 and bla_KPC-2 were more frequent in CRISPR-Cas-positive strains (p < 0.0001). Imipenem/relebactam resistance was higher in CRISPR-Cas-negative isolates. (4) Conclusions: K. pneumoniae CRISPR-Cas systems correlate with specific carbapenemase profiles, suggesting pressure against bla_OXA-48 acquisition. The coexistence of I-E and I-E* subtypes highlight synergies in targeting MGEs. CRISPR loci could be tools for subtyping organisms following MLST. Full article

Campylobacter jejuni and Campylobacter coli are the two main campylobacter species that cause foodborne campylobacteriosis. Recent studies have reported that Campylobacter spp. are prone to developing resistance to antibiotics commonly used for their treatment, with many C. coli strains identified as multidrug-resistant. This study presents the results of the whole-genome sequencing analysis of the multidrug-resistant C. coli strain BCT3 isolated in Greece from a stool specimen of a pediatric patient presenting with diarrhea. The strain was isolated using selective culture media and, based on antimicrobial susceptibility tests, was found to be resistant to ciprofloxacin, tetracycline, erythromycin, azithromycin, clarithromycin, and doxycycline. To further characterize it, we performed whole-genome sequencing, which identified strain BCT3 as C. coli. Moreover, multilocus sequence typing assigned the BCT3 to the sequence type (ST) 872, belonging to clonal complex ST-828. The presence of multiple virulence genes revealed its pathogenic potential. The detection of antimicrobial resistance genes and mutated alleles was indicative of its resistance to fluoroquinolones, macrolides, and tetracyclines, supporting the observed phenotype. To our knowledge, this is the first reported clinical case of such a multidrug-resistant C. coli strain in Greece. 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

Background: Neisseria meningitidis (N. meningitidis) is a leading cause of acute meningitis and is classified into 13 serogroups, six of which are predominantly associated with invasive meningococcal disease. This study aimed to investigate the genotype, subgenotype, and antigenic profiles of N. meningitidis serogroup B strains isolated in Vietnam. Methods: Genotyping was performed on 106 N. meningitidis strains isolated from clinical samples from Vietnamese patients and nasopharyngeal swabs of healthy adolescents between 2019 and 2024. The genetic profiles, including the porA, porB, fetA, fHbp, abcZ, adk, aroE, fumC, gdh, pdhC, and pgm genes, were analyzed using Sanger sequencing and bioinformatic methods. Results: We found that 84.9% of the strains carried VR3 families 36 or 35-1, with VR1, VR2, and VR3 families 22-25, 14, and 36 being the most prevalent. Among the 106 serogroup B isolates, 20 variants of the porB allele 3 were identified, with porB 3-1212 being the most frequent (30.2%). Dominant PorB variable loops included L1.6, L4.5, L5.7, L6.6, and L7.13. fHbp variant group 2 was predominant (104/106 strains), and 12 FetA allele variants were identified, with F1-7 being the most common (47.2%). Three clonal complexes were identified, and clonal complex ST-32 was the most predominant. Fifty-five strains (51.9%) belonged to sequence types that have not yet been assigned to any clonal complexes, and 15 strains (14.1%) with allelic profiles were not assigned to STs. The 3-253 and 3-1212 alleles of porB, the F1-7 variant of FetA, the ST-44 and ST-1576 sequence types, and the ST-41/44 complex were observed more frequently in patients compared to asymptomatic carriers, suggesting their association with more virulence. Conclusions: This study showed a high genetic and antigenic diversity of N. meningitidis serogroup B isolates in Vietnam, with VR3 family 36 most common and porB 3-1212 as the predominant allele. fHbp variant group 2 and FetA allele F1-7 were most frequent. ST-32 was the dominant clonal complex, though many strains remained unassigned, highlighting the need for ongoing molecular surveillance. Full article

Background: High-risk Escherichia coli clones, such as sequence type (ST)131 and ST1193, along with multidrug-resistant (MDR) Klebsiella pneumoniae, are globally recognized for their significant role in urinary tract infections (UTIs). This study aimed to provide an overview of the virulence factors, clonal diversity, and antibiotic resistance profiles of extended-spectrum cephalosporin (ESC)-E. coli and K. pneumoniae causing UTIs in humans in the Tebessa region of Algeria. Methods: Forty E. coli and 17 K. pneumoniae isolates exhibiting ESC-resistance were recovered (July 2022–January 2024) from urine samples of patients at three healthcare facilities to be phenotypically and genotypically characterized. Whole genome sequencing (WGS) was performed on the ST1193 clone. Results: Among K. pneumoniae isolates, all except one harbored CTX-M-15, with a single isolate carrying bla_CTX-M-194. Additionally, two K. pneumoniae isolates co-harboring bla_CTX-M-15 and bla_NDM exhibited phenotypic and genotypic hypervirulence traits. Fluoroquinolone resistance (FQR) was detected in 94.1% of K. pneumoniae isolates. The E. coli isolates carried diverse ESC-resistance genes, including CTX-M-15 (87.5%), CTX-M-27 (5%), CTX-M-1, CMY-59, and CMY-166 (2.5% each). Co-carriage of bla_ESC and bla_OXA-48 was identified in three E. coli isolates, while 62.5% exhibited FQR. Phylogenetic analysis revealed that 52.5% of E. coli belonged to phylogroup B2, including the high-risk clonal complex (CC)131 CH40-30 (17 isolates) and ST1193 (one isolate). In silico analysis of the ST1193 genome determined O75:H5-B2 (CH14-64), and the carriage of IncI1-I(Alpha) and IncF [F-:A1:B10] plasmids. Notably, core genome single-nucleotide polymorphism (SNP) analysis demonstrated high similarity between the Algerian ST1193 isolate and a previously annotated genome from a hospital in Northwest Spain. Conclusions: This study highlights the spread and genetic diversity of E. coli CC131 CH40-30 and hypervirulent K. pneumoniae clones in Algeria. It represents the first report of a CTX-M-15-carrying E. coli ST1193 in the region. The findings emphasize the urgent need for antibiotic optimization programs and enhanced surveillance to curb the dissemination of high-risk clones that pose an increasing public health threat in Algeria. A simplified method based on virulence traits for E. coli and K. pneumoniae is proposed here for antimicrobial resistance (AMR) monitoring. Full article

Background: Staphylococcus aureus constitutes a significant public health threat due to its exceptional adaptability, antimicrobial resistance (AMR), and capacity to form biofilms, all of which facilitate its persistence in clinical and environmental settings. Methods: This study undertook an extensive in silico analysis of 44,069 S. aureus genomic sequences acquired from the NCBI database to assess the global distribution of biofilm-associated and resistance-associated genes. The genomes were categorized into human clinical and environmental groups, with clinical samples representing a predominant 96%. Results: The analysis revealed notable regional discrepancies in sequencing efforts, with Europe and North America contributing 76% of the genomes. Key findings include the high prevalence of the ica locus, which is associated with biofilm formation, and its robust correlation with other genes, such as sasG, which was exclusively linked to SCCmec type IIa. The AMR gene analysis revealed substantial genetic diversity within environmental samples, with genes like vga(E) and erm being identified as particularly prominent. The clonal complex analysis revealed ST8 (USA300) and ST5 as the predominant types in human clinical isolates, while ST398 and ST59 were most frequently observed in environmental isolates. SCCmec type IV was globally prevalent, with subtype Iva being strongly associated with ST8 in North America and subtype IVh with ST239 in Europe. Conclusions: These findings underscore the dynamic evolution of S. aureus via mobile genetic elements and highlight the necessity for standardized metadata in public genomic databases to improve surveillance efforts. Furthermore, they reinforce the critical need for a One Health approach in monitoring S. aureus evolution, particularly concerning the co-dissemination of biofilm and resistance genes across various ecological niches. Full article

Urinary tract infections (UTIs) are a leading cause of illness in children and adults of all ages, with uropathogenic Escherichia coli (UPEC) being the primary agent responsible. During colonization and subsequent infection of the urinary tract (UT), UPEC requires the expression of genes associated with virulence, such as those that encode the fimbrial adhesins FimH, PapG, and CsgA, as well as the presence of the TosA protein and the flagellar appendages of the bacteria. However, for colonization and infection to be successful, UPEC must overcome the host’s immunological barriers, such as physical barriers, expressed peptides and proteins, and immune cells found in the UT. In this context, the UT functions as an integral system where these factors act to prevent the colonization of uropathogens. Significant genetic diversity exists among UPEC strains, and the clonal complex ST131 represents one of the key lineages. This lineage has a high content of virulence genes, multiple mechanisms of antibiotic resistance, and a high frequency of extended-spectrum β-lactamases (ESBLs). New knowledge regarding protein structures known as adhesins and their role in the infection process can help identify therapeutic targets and aid in the design of vaccines. These vaccines could be based on the development of chimeric fusion proteins (FimH + CsgA + PapG), which may significantly reduce the incidence of UTIs in pediatric and adult patients. Full article

Listeria monocytogenes is an important foodborne pathogen, markedly persistent even in harsh environments and responsible for high hospitalization and mortality rates. The aim of the present study was to detect the strains circulating in Sicily over a five-year period and characterize their antimicrobial resistance profiles. The key element of this study was the sharing of data among various entities involved in food control and clinical surveillance of listeriosis in order to develop an integrated approach for this pathogen. A total of 128 isolates were analyzed, including 87 food-source strains and 41 clinical specimens. Whole-genome sequencing (WGS) was performed for sequence type (ST) and clonal complex (CC) identification through multilocus sequence typing (MLST) analysis. Antimicrobial resistance was assessed using the Kirby–Bauer method. The majority of strains belonged to serotype IVb (34/41 and 53/87 of clinical and food-source isolates, respectively) and were subtyped as CC2-ST2 (28/34 and 41/53 of clinical and food-source isolates respectively). Most of the isolates were susceptible to the main antimicrobials recommended for treatment of listeriosis. Resistance (R) and intermediate resistance (I) percentages worthy of attention were found against oxacillin (R: 85.9%) and clindamycin (I: 34.6%) in the food-source isolates and trimethoprim/sulfamethoxazole (R: 29.23%) in the clinical isolates. Also, 7.7% of the food-source isolates were multidrug resistant. Our results highlight how the punctual comparison between food and clinical strains is an essential tool for effectively tracking and preventing foodborne outbreaks. Full article

Background/Objectives: Acinetobacter baumannii is an increasingly significant nosocomial pathogen causing severe infections globally. The emergence of multidrug-resistant A. baumannii strains has raised concerns about the efficacy of current treatment options. This study aimed to investigate the molecular epidemiology and antimicrobial resistance patterns of A. baumannii isolates from Kazakhstan. Methods: We collected nine A. baumannii isolates in 2022–2023 in Karaganda, Kazakhstan, which were then subjected to whole-genome sequencing (WGS) using the IonTorrent platform for genome characterization. Multilocus sequence typing (MLST) was used to classify the isolates into distinct clonal complexes. In addition, antibiotic susceptibility testing was conducted using the standard methods for a range of antibiotics commonly used against A. baumannii. Results: Our results revealed a high degree of genomic diversity among isolates from Kazakhstan, with multiple distinct classes identified: ST78 (n = 4, 44.4%), ST15 (n = 2, 22.2%), ST2 (n = 2, 22.2%), and ST193 (n = 1, 11%). MLST analysis showed that ST78^Pas/1104^Oxf (harboring blaOXA-72 and blaOXA-90 genes) were prevalent among the multidrug-resistant isolates. Based on the results of MLST, KL, and OCL, the analyzed isolates were assigned to specific international clones: IC2—ST2(Pas)-KL2/168-OCL1, IC4—ST15(Pas)-KL9-OCL7, and IC6—ST78(Pas)-KL49-OCL1. Notably, these isolates exhibited resistance to multiple antibiotics including meropenem, imipenem, gentamicin, amikacin, and ciprofloxacin. Conclusions: This study highlighted the complex molecular epidemiology of A. baumannii in Kazakhstan over a two-year period, underscoring the need for targeted surveillance strategies to monitor antimicrobial resistance patterns. The emergence and dissemination of multidrug-resistant strains within this timeframe emphasizes the importance of whole-genome sequencing as a diagnostic tool and underscores the challenges posed by these infections. Full article

Streptococcus pneumoniae serotype 1 is one of the most prevalent serotypes commonly associated with invasive pneumococcal disease cases and outbreaks worldwide. Several sequence types of this serotype have been identified globally, including those exhibiting both high virulence potential and antimicrobial resistance profiles. This systematic review presents the global distribution of clones of pneumococcal serotype 1, describing their circulating patterns in various regions in the world. A database search was conducted in Google Scholar, PubMed, Scopus, ScienceDirect, and Web of Science using keywords related to Streptococcus pneumoniae serotype 1. The inclusion criteria entailed peer-reviewed studies published in English describing the utilization of at least one molecular genotyping tool to identify S. pneumoniae serotype 1 clones based on their sequence types. Data extracted were managed and analyzed using Microsoft Excel 365 (Version 2108). Forty-three studies were finally included in the systematic review. A total of 103 MLST serotype 1 sequence types were identified in 48 countries. These clones were widely reported to be associated with invasive pneumococcal diseases. Globally, ST217 and ST306 clonal complexes (CC217 and CC306) were the predominant lineages of serotype 1 sequence types, exhibiting distinct continental distribution patterns. CC217, characterized by ST217, ST303, ST612, ST618, and ST3081, was predominant in Africa and Asia. ST306 clonal complex, which is grouped into ST306, ST304, and ST227 were mostly found in Europe, Oceania, North America, and some countries in South America. ST615 was predominant in Chile, Peru, and Argentina. The hypervirulence nature of serotype 1, coupled with its complex genetic diversity, poses a significant public health threat. Our findings emphasize the need for enhanced surveillance and targeted interventions to mitigate the spread of these hypervirulent clones, ultimately informing evidence-based strategies for disease prevention and control. Full article