Search Results (135)

Chagas disease (ChD) caused by Trypanosoma cruzi remains a major public health concern, affecting approximately 8 million people worldwide. However, the number of undiagnosed cases is likely much higher. Existing treatments rely on benznidazole and nifurtimox which, despite their efficacy during the acute phase of infection, are often associated with severe side effects that can be life-threatening. As a promising alternative, actinomycetes—which are renowned for producing pharmacologically and industrially relevant metabolites—have demonstrated potent antimicrobial properties; however, their antiparasitic potential remains largely unexplored. This study evaluated the anti-trypanocidal activities of extracellular metabolites produced by Streptomyces thermocarboxydus strain Chi-43 (ST-C43) and Streptomyces sp. strain Chi-104 (S-C104) against epimastigote, trypomastigote, and amastigote forms of T. cruzi. The strains were cultured in ISP2 broth, and their extracellular metabolites were assessed via antiparasitic diffusion assays in microplates. The 50% lethal concentration (LC₅₀) values ranged from 102 to 116 μg/mL against epimastigotes and trypomastigotes. The antiparasitic activity was confirmed through 3-(4,5-dimetiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based spectrophotometric assays and optical microscopy. Toxicity assays revealed that the extracellular metabolites were non-toxic to Artemia salina, non-cytotoxic to Huvecs, and non-hemolytic to human erythrocytes. Dose–response regression analysis showed statistically significant differences (p ≤ 0.05). LC-MS/MS analysis identified amphomycin and K-252c aglycone staurosporine as the active antiparasitic compounds. These findings highlight the potential of Streptomyces-derived extracellular metabolites as novel, selective, and safe anti-T. cruzi agents. Nevertheless, further studies in murine or preclinical models are needed to validate their efficacy and support future clinical applications for the treatment of ChD. 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

Carbapenemase-producing Klebsiella pneumoniae is responsible for multiple serious infections with high mortality rates. K. pneumoniae carbapenemases (KPCs) are the most commonly isolated carbapenemases worldwide. To study the epidemiological and molecular characteristics of KPC-producing K. pneumoniae (KPC-KP), we conducted a retrospective study at the University General Hospital of Ioannina, Greece. A total of 177 K. pneumoniae clinical strains from the period 2014–2015 were confirmed as KPC producers by polymerase chain reaction (PCR) and were further examined for the presence of bla_VIM, bla_NDM, bla_TEM, bla_SHV, and bla_CTX-M genes. Using the amplification refractory mutation system (ARMS) method, we identified the presence of the KPC-2 allele in 130 strains and the KPC-9 allele in 47. Strains from both allele groups belonged to the sequence type 258 (ST258). KPC-9 was responsible for a distinct outbreak, considered part of the broader KPC-2 outbreak. Molecular characterization of selected KPC-KP isolates from the period 2021–2022 revealed their continued presence in our hospital. Comparison of the antimicrobial susceptibility profiles of the two alleles showed a statistically significant increase in minimum inhibitory concentration (MIC) for ceftazidime (p = 0.03) and higher resistance to amikacin (p = 0.012) and colistin (p < 0.001) for KPC-9 compared to the KPC-2 allele. The two KPC alleles had similar mortality rates. This study demonstrates the heterogeneity of resistance genes in carbapenem-resistant K. pneumoniae (CR-KP) within a single-hospital setting and underscores the need for immediate containment measures. Full article

Background/Objectives: Salmonella is a major cause of foodborne illnesses, with multidrug-resistant (MDR) strains posing significant threats to public health worldwide. This study investigated the prevalence and antimicrobial resistance (AMR) of Salmonella, focusing on extended-spectrum β-lactamase (ESBL)-producing Salmonella in retail poultry meat in Korea. Methods: A total of 300 poultry meat samples were collected nationwide from retail markets. Multi-locus sequence typing, serotyping, and antimicrobial susceptibility testing were performed. Whole-genome sequencing (WGS) analysis was conducted against 28 representative ESBL-producing S. Infantis isolates to identify the genetic characteristics and phylogenetic relationship. Results: Salmonella was detected in 81.3% of raw poultry meat samples, with S. Infantis ST32 being the dominant serotype in chicken (53.0%) and S. Typhimurium ST19 predominant in duck (39.0%). MDR was identified in 58.2% of samples, with a significantly higher rate in chicken isolates than in duck isolates (p < 0.001). Notably, 75.3% of chicken MDR isolates were ESBL-producing S. Infantis carrying bla_CTX-M-65. WGS of 28 geographically and phenotypically representative ESBL-producing S. Infantis revealed five clonal clusters, suggesting the widespread dissemination of ESBL-producing S. Infantis across Korea’s poultry supply chain. All 28 ESBL-producing S. Infantis isolates contained a pESI-like megaplasmid, carrying multiple resistance and virulence genes, with sequences highly identical to plasmids reported in the United States, indicating potential international transmission. Conclusions: This study emphasizes the urgent need for continuous surveillance and responsible antibiotic use in livestock under a One Health framework. WGS can provide an effective tool for tracking AMR evolution and clonal spread within and across regions. Full article

Background: Antimicrobial resistance (AMR) has become a serious global challenge in the 21st century. Poultry, including turkeys, are a vital source of animal-derived protein worldwide. Commensal bacterial strains in poultry can act as reservoirs for AMR, making monitoring them crucial for both veterinary and public health. Enterococcus species are emerging pathogens, particularly in severe nosocomial infections. Methods: This study aimed to assess the resistance profiles of commensal Enterococcus strains isolated (n = 470) from large-scale turkey flocks in Hungary. From each animal, two swab samples were collected: one from the oropharyngeal region near the tracheal entrance and one from the cloaca. The samples were subsequently processed, and the minimum inhibitory concentration (MIC) was determined following the Clinical and Laboratory Standards Institute (CLSI) guidelines. The tested antibiotics included amoxicillin, amoxicillin–clavulanic acid, imipenem, neomycin, doxycycline, florfenicol, tylosin, enrofloxacin, potentiated sulfonamide, vancomycin, ceftriaxone, spectinomycin, tiamulin, lincomycin, and colistin. The dilution range for MIC determination was set between 512 and 0.001 µg/mL. Results: Resistance to amoxicillin, a first-line treatment for Enterococcus infections, was low (11.1%). However, high resistance levels were observed for tylosin (62.6%), florfenicol (51.1%), doxycycline (48.7%), and enrofloxacin (45.5%). Notably, vancomycin resistance reached 15.5%, a finding consistent with global trends. Compared to human-derived Enterococcus data, resistance to aminopenicillins was significantly lower in turkey isolates, while neomycin resistance levels were comparable to those observed in human E. faecalis strains. Conclusions: The findings underscore the necessity of continuous surveillance of AMR trends in poultry production. While amoxicillin remains an effective treatment, the presence of multidrug-resistant strains and vancomycin-resistant isolates raises concerns regarding the potential dissemination of resistance genes. Future studies should incorporate next-generation sequencing to elucidate the genetic mechanisms underlying resistance. Additionally, integrating antibiotic usage data from farms may provide further insights into resistance dynamics. Strengthening antibiotic stewardship programs and fostering collaboration between veterinary and human medicine are crucial steps in addressing AMR under the One Health framework. Full article

Crude oil and its derivates are among the most important environmental pollutants, where P. aeruginosa strains producing AlkB1 and AlkB2 alkane hydroxylases are often involved in their biodegradation. The aim of this study was to analyze antibiotic resistance and virulence determinants of a P. aeruginosa isolate cultured from a hydrocarbon-contaminated soil sample from Ogoniland, Nigeria, and to compare its characteristics with P. aeruginosa isolates cultured worldwide from hydrocarbon-contaminated environments or from clinical samples. Using the ResFinder reference database, a catB7 chloramphenicol acetyltransferase gene, an ampC-type PDC β-lactamase gene, and an OXA-50 type β-lactamase gene were identified in all P. aeruginosa strains analyzed in this study. In some of these P. aeruginosa strains, loss-of-function mutations were detected in the regulatory genes mexR, nalC, or nalD, predicting an efflux-mediated acquired antibiotic-resistance mechanism. Several P. aeruginosa sequence types that were associated with oil-contaminated environments have also been cultured from human clinical samples worldwide, including sequence types ST532, ST267, ST244, and ST1503. Our findings also indicate that environmental P. aeruginosa may serve as the source of human infections, warranting further studies from a One Health perspective about the application of P. aeruginosa for the in situ bioremediation of hydrocarbon-contaminated sites. Full article

(1) Background. Although Arum spp. are toxic in their raw state, they are sometimes used as food within their native ranges. (2) Methods. We review the available literature in order to provide an overview of its use and detoxification procedures worldwide. (3) Results. The food use of lords-and-ladies was already mentioned by Theophrastus, Dioscorides, Matthioli, Durante, Gerard, and Sirennius. In the references concerning 19th–21st-century use, seven species were identified: A. cyrenaicum, A. discoridis, A. italicum, A. maculatum, A. orientale, A. palaestinum, and A. rupicola. Past or current culinary use of the plant has been recorded in Morocco, Libya, the United Kingdom, the Scilly Islands, Germany, Switzerland, Italy, Romania, Ukraine (including Crimea), Czechia, Slovenia, Croatia, Bosnia-Herzegovina, Albania, Georgia, Türkiye, Syria, Palestine, Lebanon, Israel, Iraq, and Iran. (4) In Europe, rhizomes were used, mainly as a famine food. In SW Asia, the aerial parts remain an important element of local cuisine. Several detoxification procedures are used before consumption, such as prolonged boiling, often involving straining the boiled water and lowering the pH with lemon juice, sumac, citric acid, sorrel leaves, or pomegranate juice. (5) Conclusions. Further studies are needed to assess the safety of Arum use and record traditional local recipes in SW Asia. Full article

Enterotoxigenic Escherichia coli (ETEC) is a major cause of diarrheal disease worldwide, particularly in children in low- and middle-income countries. Its ability to rapidly colonize the intestinal tract through diverse colonization factors and toxins underpins its significant public health impact. Despite extensive research and several vaccine candidates reaching clinical trials, no licensed vaccine exists for ETEC. This review explores the temporal and spatial coordination of ETEC virulence factors, focusing on the interplay between adherence mechanisms and toxin production as critical targets for therapeutic intervention. Advancements in molecular biology and host–pathogen interaction studies have uncovered species-specific variations and cross-reactivity between human and animal strains. In particular, the heat-labile (LT) and heat-stable (ST) toxins have provided crucial insights into molecular mechanisms and intestinal disruption. Additional exotoxins, such as EAST-1 and hemolysins, further highlight the multifactorial nature of ETEC pathogenicity. Innovative vaccine strategies, including multiepitope fusion antigens (MEFAs), mRNA-based approaches, and glycoconjugates, aim to enhance broad-spectrum immunity. Novel delivery methods, like intradermal immunization, show promise in eliciting robust immune responses. Successful vaccination against ETEC will offer an effective and affordable solution with the potential to greatly reduce mortality and prevent stunting, representing a highly impactful and cost-efficient solution to a critical global health challenge. Full article

Background/Objectives: Antimicrobial resistance of the E. coli O25 ST131 clonal lineage poses a significant therapeutic challenge worldwide, often involving resistance to fluoroquinolones and extended-spectrum beta-lactamase (ESBL) production. This retrospective study compared the dissemination of multidrug-resistant E. coli O25 ST131 isolated from the urine of outpatients at the largest Croatian clinical microbiology department across six years over two study periods. Methods: The E. coli O25 ST131 clonal lineage was detected via a rapid PCR method using pabB and trpA primers after positive agglutination with E. coli serogroup O25 antisera. ESBL phenotypes and antibiotic susceptibility were investigated according to EUCAST guidelines and breakpoint tables. Results: In the first period, there were a total of 45 isolates of E. coli O25 ST131, among which 30 were isolates with proven ESBL production. In the second period, a total of 114 isolates of E. coli O25 ST131 were detected, among which 75 (65.8%) were ESBL-positive (p > 0.05). In ESBL-negative strains, the multidrug-resistant (MDR) phenotype was characterized by simultaneous resistance to ampicillin, co-trimoxazole, and fluoroquinolones (with an equal proportion of 3/15 isolates in the first period and 7/39 isolates in the second period, p > 0.05). There was no statistically significant difference in the frequency of MDR detection across the two study periods (36/45 and 98/114, p > 0.05). This is the first detection of E. coli O25 ST131 in the outpatient population in Zagreb. Conclusions: There was no statistically significant difference in the frequency of detecting the E. coli O25 ST 131 clone across the two study periods. The high frequency of MDR phenotype among ESBL-negative isolates of E. coli O25 ST131 and an equally high proportion of MDR strains among ESBL producers in this clonal lineage, with the total detection of MDR isolates ≥ 80% in both study periods, are the reasons why this bacterial clone poses a public health threat and why further investigation into its metabolic and virulence characteristics is needed in order to estimate its spreading potential among the outpatient population in Zagreb. Full article

Multidrug-resistant Acinetobacter baumannii is a major concern in healthcare institutions worldwide. Several reports described the dissemination of A. baumannii high-risk clones that are responsible for a high number of difficult-to-treat infections. In our study, 19 multidrug-resistant A. baumannii strains from Budapest, Hungary, were investigated based on whole-genome sequencing (WGS). The obtained results were analysed together with data from 433 strains of A. baumannii from the Pathogenwatch database. WGS analysis of 19 A. baumannii strains detected that 12 belonged to ST2 and seven belonged to ST636. Among ST2 strains, 11 out of 12 carried either bla_OXA-23 or bla_OXA-58 genes; however, all strains of ST636 uniformly carried bla_OXA-72 gene. All strains of ST2 and ST636 carried bla_OXA-66 and bla_ADC-25 genes. Based on core genome multilocus sequence typing (cgMLST), 10 strains of ST2 belonged to cgMLST906, one strain to cgMLST458, and one strain to cgMLST1320; by contrast, all strains of ST636 belonged to cgMLST1178. Certain virulence determinants were present in all strains of both ST2 and ST636, namely, Ata, Bap, BfmRS, T2SS and PNAG. Interestingly, OmpA was present in all strains of ST2, but it was absent in all strains of ST636. Comparative analysis of 19 strains of this study and the collection of 433 isolates from Pathogenwatch database, proved a diverse clonal distribution of high-risk A. baumannii clones in Europe. The major clone in Europe is ST2, which is present all over the continent. However, ST636 has been mainly reported in Eastern Europe. Interestingly, cgMLSTs of ST2 correspond to the production of different beta-lactamases, namely, OXA-82 in cgMLST116, OXA-72 in cgMLST506, and cgMLST556, PER-1 in cgMLST456 and cgMLST1041. Our study demonstrates that the ST2 high-risk clone of A. baumannii is the most widespread in Europe; however, based on cgMLST analysis, a detailed detection of beta-lactamase production can be determined. Full article

Background/Objectives: Bloodstream infection by carbapenem-resistant Acinetobacter baumannii (CRAB) is a serious clinical problem worldwide. To study its clonal relationship and genetic features, we report the draft genome sequence of CRAB strains isolated from human blood in South Korea. Methods: Among A. baumannii strains isolated from patients at nine general hospitals in 2020, 12 CRAB strains of different genotypes were selected. Genomic DNA was sequenced using a combination of Illumina MiSeq and Oxford Nanopore MinION platforms. Antimicrobial susceptibility testing was performed using the disk diffusion method. Antimicrobial resistance and virulence genes were investigated in silico using the Center for Genomic Epidemiology server and the Virulence Factors Database. Results: The multilocus sequence types of isolates included ST191, ST195, ST357, ST369, ST451, ST469, ST491, ST784, ST862, ST1933, ST2929, and a novel type, ST3326. The predominant sequence type, ST191, demonstrated close genetic relationships with several isolates, including ST469, ST369, ST195, ST784, ST491, and ST3326, with ST3326 classified as a subgroup of ST191. We found 18 antimicrobial resistance genes and one quaternary ammonium compound resistance gene. All examined strains harbored bla_OXA-23, which is associated with carbapenem resistance. While variations in antibiotic and disinfectant resistance genes were observed, all isolates exhibited similar virulence factors, with the exception of the biofilm and capsule production genes. Conclusions: This nationwide report of the draft genome sequence of patient-derived strains provides valuable insights into the genomic features associated with clonal relationships and antimicrobial resistance of CRAB in bloodstream infections. Full article

Streptococcus agalactiae is an important pathogen responsible for cases of high mortality in farmed and wild fish worldwide. In Brazil, this bacterium has been commonly associated with outbreaks in Nile tilapia farms, but other native fish species are also susceptible. Since floating cages are one of the most common culture systems used in the country, the close contact between farmed tilapia and native fish species presents a risk concerning the transmission of this pathogen. In this study, we characterized a mortality outbreak in free-living trahira and in farmed arapaima, as well as the genetic and antimicrobial susceptibility patterns of the isolates obtained. During the outbreaks, moribund fish were sampled and subjected to bacterial examination, after which the isolates were identified via MALDI-ToF analysis. Genotyping was evaluated using repetitive sequence-based PCR (REP-PCR) and multilocus sequence typing (MLST). Antimicrobial susceptibility was evaluated using disc diffusion assays. In addition, whole-genome analysis also was performed. S. agalactiae was identified in all diseased fish, all of which belonged to serotype Ib; however, trahira strains were classified as non-typeable lineages in the MLST assay, while arapaima strains were classified as ST260. These isolates were shown to be similar to the main genotype found in Nile tilapia in Brazil, using REP-PCR, MLST and phylogenomic analysis. The pathogenicity of the bacterium was confirmed by Koch’s postulates for both fish species. The antimicrobial susceptibility assay showed variable results to the same antibiotics among the isolates, prompting four of the isolates to be classified as multidrug-resistant. This study represents the first report of a natural outbreak of Streptococcus agalactiae infection in wild trahira and farmed arapaima inhabiting the same aquatic environment as Nile tilapia. Full article

Background/Objectives: The emergence and dissemination of carbapenem-resistant organisms, particularly Acinetobacter baumannii and Klebsiella pneumoniae, pose a significant threat to healthcare systems worldwide. This retrospective study aims to characterise carbapenem-resistant Acinetobacter baumannii (CRAB) and carbapenem-resistant Klebsiella pneumoniae (CRKP) strains in a teaching hospital and to determine the risk factors associated with patients’ in-hospital mortality. Methods: A total of 90 CRAB and 63 CRKP were included in this study. Carbapenemase genes and MLST types of CRAB and CRKP were determined using specific primers. Risk factors associated with in-hospital mortality were analysed with collected data. Results: All the CRAB strains consisted of OXA carbapenemase genes, with 98% of the strains co-harbouring blaOXA-23-like and blaOXA-51-like carbapenemase genes. Conversely, blaNDM is the predominant carbapenemase gene in CRKP, followed by blaOXA-48-like carbapenemase genes. ST2 and ST20 are the dominant MLST types in CRAB and CRKP, respectively. In CRAB, multivariate analysis identified age, ethnicity, the presence of a mechanical ventilator, and patients who experienced previous exposure to clindamycin in the last 90 days as associated with an increased risk of in-hospital mortality. In contrast, older age, male, ICU admission, and the presence of an indwelling urinary catheter were significantly associated with an increased risk of mortality for patients with CRKP. Conclusions: Both CRAB and CRKP lead to high rates of mortality. The MLST profile showed that the genomic patterns of CRKP were highly diverse, whereas CRAB strains had low genetic diversity. To tackle these challenging pathogens, robust surveillance and an in-depth understanding of molecular epidemiology and genomics studies are needed to tailor infection control strategies and individualise treatment approaches. Full article

Throughout Mycobacterium leprae’s (M. leprae) evolutionary trajectory, nearly half of its genome was converted into pseudogenes. Despite this drastic reduction in genetic content, the genome sequence identity among M. leprae isolates worldwide is remarkably high compared to other pathogens. In this study, we investigated the genotype and morphotype of three M. leprae strains: the reference strain Thai-53 (genotype 1A), and two clinical isolates from Brazilian leprosy relapse patients, which were Br014-03 (genotypes 3I) and Br014-01(4N). We compared their genome sequences and their interaction with human Schwann cells from the ST88-14 lineage and with human primary macrophages. On the genetic level, we observed over a hundred missense mutations in the three strains, translated into significant phenotypic changes such as: prolonged doubling time, altered cytokine induction, reduced interaction rates, and decreased intracellular viability in Schwann cells. Our findings underscore the concept that despite their 99.992% identity, even small genomic disparities in M. leprae genomes can elicit substantial alterations in bacilli interaction with host cells and subsequent immune responses. Consequently, our data could lead to better comprehension of correlation between pathogen mutations and the diverse clinical manifestations observed in leprosy patients. Full article

Entomopathogenic fungi (EPFs) are capable of infecting a variety of insect pests and are widely used as biopesticides worldwide. This study intended to isolate indigenous EPFs from cadavers of Protaetia brevitarsis and investigate their effects on the fall armyworm Spodoptera frugiperda (FAW), a globally widespread invasive pest. Morphological and molecular analyses confirmed four native EPF strains all belong to Beauveria bassiana. Pathogenicity tests showed they were virulent toward FAW 1st instar larvae. The application of EPFs either by dipping or spraying significantly increased the larval mortalities compared to the control group, with corrected mortalities ranging from 92% to 73% after dipping in a fungal suspension of 10⁸ conidia/mL, and those ranging from 76% to 35% after spraying with a fungal suspension of 10⁷ conidia/mL. Our findings revealed the infectivity of four strains to FAW larvae significantly changed in a dose- and time-dependent manner. In addition, the combination use of the local B. bassiana strain and parasitoid Microplitis prodeniae resulted in a significantly enhanced S. frugiperda 3rd instar larval mortality compared to a single inoculation with one of them, suggesting an apparent synergistic effect for the joint application of these two biological control agents. The mortality inflicted by B. bassiana was probably promoted by the release of parasitoids, since the parasitoids’ movements and attacks could strengthen the fungal distribution and infection processes. This study underscores the potential of the combination use of EPFs and parasitoids against S. frugiperda early-instar larvae, and provides insights into the consequences of integrating these EPFs into integrated pest management systems. Full article