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BioMed Research International 2019The emergence of antimicrobial-resistant and virulent enterococci is a major public health concern. While enterococci are commonly found in food of animal origin, the...
The emergence of antimicrobial-resistant and virulent enterococci is a major public health concern. While enterococci are commonly found in food of animal origin, the knowledge on their zoonotic potential is limited. The aim of this study was to determine and compare the antimicrobial susceptibility and virulence traits of and isolates from human clinical specimens and retail red meat in Slovenia. A total of 242 isolates were investigated: 101 from humans (71 , 30 ) and 141 from fresh beef and pork (120 , 21 ). The susceptibility to 12 antimicrobials was tested using a broth microdilution method, and the presence of seven common virulence genes was investigated using PCR. In both species, the distribution of several resistance phenotypes and virulence genes was disparate for isolates of different origin. All isolates were susceptible to daptomycin, linezolid, teicoplanin, and vancomycin. In both species, the susceptibility to antimicrobials was strongly associated with a food origin and the multidrug resistance, observed in 29.6% of and 73.3% clinical isolates, with a clinical origin (Fisher's exact test). Among meat isolates, in total 66.0% of and isolates were susceptible to all antimicrobials tested and 32.6% were resistant to either one or two antimicrobials. In , several virulence genes were significantly associated with a clinical origin; the most common (31.0%) gene pattern included all the tested genes except . In meat isolates, the virulence genes were detected in only and the most common pattern included , , and (32.5%), of which showed a statistically significant association with a clinical origin. These results emphasize the importance of in red meat as a reservoir of virulence genes involved in its persistence and human infections with reported severe outcomes.
Topics: Animals; Anti-Bacterial Agents; Drug Resistance, Bacterial; Enterococcus faecalis; Enterococcus faecium; Food Microbiology; Humans; Pork Meat; Slovenia
PubMed: 31211134
DOI: 10.1155/2019/2815279 -
Journal of Microbiology and... Feb 2023A cryptic plasmid (pTH32) was characterized from 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of...
A cryptic plasmid (pTH32) was characterized from 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). and are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. and encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel shuttle vector, pMJ32E (7.3 kb, Em), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in 29212 and 31 but not in other LAB species. A gene, encoding aminopeptidase A (PepA) from CY54, was successfully expressed in 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than 31 cell (control). When 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of and species.
Topics: Plasmids; Genetic Vectors; Enterococcaceae; DNA Replication; Proteins; Open Reading Frames
PubMed: 36575862
DOI: 10.4014/jmb.2209.09024 -
Journal of Immunology Research 2021This study included 74 Chinese male patients with HCC. They were divided into early ( = 19), intermediate ( = 37), and terminal ( = 18) groups, referred to as Barcelona... (Observational Study)
Observational Study
METHOD
This study included 74 Chinese male patients with HCC. They were divided into early ( = 19), intermediate ( = 37), and terminal ( = 18) groups, referred to as Barcelona Clinic Liver Cancer stage 0+A, B, and C+D, respectively. Paired fecal and plasma samples were collected. Microbial composition and profiles were analyzed by 16S rRNA gene sequencing. The levels of gut damage marker (regenerating islet-derived protein 3 (REG3)) and microbial translocation markers (soluble CD14 (sCD14), lipopolysaccharide-binding protein (LBP), peptidoglycan recognition proteins (PGRPs)) were determined in plasma samples of patients by ELISA. Twenty plasma cytokine and chemokines were determined by Luminex.
RESULTS
In early, intermediate, and terminal groups, the abundance of the family decreased significantly (3.52%, 1.55%, and 0.56%, respectively, = 0.003), while the abundance of the family increased significantly (1.6%, 2.9%, and 13.4%, respectively, = 0.022). Levels of REG3 and sCD14 were markedly elevated only in the terminal group compared with the early ( = 0.025 and = 0.048) and intermediate groups ( = 0.023 and = 0.046). The level of LBP significantly increased in the intermediate ( = 0.035) and terminal ( = 0.025) groups compared with the early group. The PGRP levels were elevated only in the terminal group compared with the early group ( = 0.018). The ratio of to was significantly associated with the levels of REG3, LBP, sCD14, and PGRPs. With HCC progression, increased levels of inflammatory cytokines accompanied by a T cell-immunosuppressive response and microbial translocation were observed.
CONCLUSION
Gut microbiota compositional and functional shift, together with elevated gut damage and microbial translocation, may promote HCC development by stimulating inflammatory response and suppressing T cell response.
Topics: Actinobacteria; Aged; Bacterial Translocation; Carcinoma, Hepatocellular; DNA, Bacterial; Disease Progression; Dysbiosis; Enterococcaceae; Feces; Gastrointestinal Microbiome; Humans; Liver Neoplasms; Male; Middle Aged; RNA, Ribosomal, 16S
PubMed: 34722779
DOI: 10.1155/2021/4973589 -
MSphere Nov 2020Vancomycin-resistant (VRE) is a leading cause of hospital-acquired infections and continues to spread despite widespread implementation of pathogen-targeted control...
Vancomycin-resistant (VRE) is a leading cause of hospital-acquired infections and continues to spread despite widespread implementation of pathogen-targeted control guidelines. Commensal gut microbiota provide colonization resistance to VRE, but the role of gut microbiota in VRE acquisition in at-risk patients is unknown. To address this gap in our understanding, we performed a case-control study of gut microbiota in hospitalized patients who did (cases) and did not (controls) acquire VRE. We matched case subjects to control subjects by known risk factors and "time at risk," defined as the time elapsed between admission until positive VRE screen. We characterized gut bacterial communities using 16S rRNA gene amplicon sequencing of rectal swab specimens. We analyzed 236 samples from 59 matched case-control pairs. At baseline, case and control subjects did not differ in gut microbiota when measured by community diversity ( = 0.33) or composition ( = 0.30). After hospitalization, gut communities of cases and controls differed only in the abundance of the -containing operational taxonomic unit (OTU), with the gut microbiota of case subjects having more of this OTU than time-matched control subjects ( = 0.01). Otherwise, case and control communities after the time at risk did not differ in diversity ( = 0.33) or community structure ( = 0.12). Among patients who became VRE colonized, those having the -containing OTU on admission had lower relative abundance once colonized ( = 0.004). Our results demonstrate that the 16S profile of the gut microbiome does not predict VRE acquisition in hospitalized patients, likely due to rapid and profound microbiota change. The gut microbiome does not predict VRE acquisition, but it may be associated with expansion, suggesting that these should be considered two distinct processes. The Centers for Disease Control and Prevention estimates that VRE causes an estimated 54,000 infections and 539 million dollars in attributable health care costs annually. Despite improvements in hand washing, environmental cleaning, and antibiotic use, VRE is still prevalent in many hospitals. There is a pressing need to better understand the processes by which patients acquire VRE. Multiple lines of evidence suggest that intestinal microbiota may help some patients resist VRE acquisition. In this large case-control study, we compared the 16S profile of intestinal microbiota on admission in patients that did and did not subsequently acquire VRE. The 16S profile did not predict subsequent VRE acquisition, in part due to rapid and dramatic change in the gut microbiome following hospitalization. However, spp. present on admission predicted decreased abundance after VRE acquisition, and spp. present on admission predicted dominance after VRE acquisition. Thus, VRE acquisition and domination may be distinct processes.
Topics: Anti-Bacterial Agents; Bacterial Physiological Phenomena; Case-Control Studies; Cross Infection; Drug Resistance, Bacterial; Enterococcus faecium; Feces; Female; Gastrointestinal Microbiome; Gram-Positive Bacterial Infections; Humans; Intensive Care Units; Male; Middle Aged; RNA, Ribosomal, 16S; Vancomycin; Vancomycin-Resistant Enterococci
PubMed: 33208515
DOI: 10.1128/mSphere.00537-20 -
Medical Principles and Practice :... 2021The purpose of this study was to investigate the distribution pattern of genes responsible for erythromycin and tetracycline resistance and their association with...
OBJECTIVES
The purpose of this study was to investigate the distribution pattern of genes responsible for erythromycin and tetracycline resistance and their association with resistance phenotypes in enterococcus isolates.
MATERIALS AND METHODS
Eighty-six Enterococcus faecalis and 26 E. faecium isolates were collected from 2 hospitals in Kerman, Iran. Minimum inhibitory concentration of erythromycin and tetra-cycline was determined and then genes encoding resistance to erythromycin - erm (A-C), mef, and msr - and tetracycline - tet (M), tet (O), tet (S), tet (K), and tet (L) - were investigated.
RESULTS
In all resistant isolates (n = 72, 64%), high-level resistance to both tested antibiotics was found. The most prevalent erm gene was erm (B) (77.7%), followed by erm (A) (15.2%) and erm (C) (8.3%). Genes mediating erythromycin efflux were detected in 70.8% (mef) and 9.7% (msr) of resistant isolates. Regarding tetracycline, tet (M) was detected at the highest rate (50%), followed by tet (O) (31%) and tet (S) (11%). Export of tetracycline was found in 31% (tet (K)) and 12% (tet (L)) of isolates.
CONCLUSION
A high prevalence of high-level resistance to both erythromycin and tetracycline was documented. Alterations at the ribosomal level was more frequently detected in erythromycin and tetracycline resistance than efflux systems. Concurrent resistance mechanisms were more involved in resistance to erythromycin than tetracycline.
Topics: Anti-Bacterial Agents; Drug Resistance, Microbial; Enterococcus; Enterococcus faecalis; Enterococcus faecium; Erythromycin; Gram-Positive Bacterial Infections; Humans; Iran; Microbial Sensitivity Tests; Tetracyclines
PubMed: 33789292
DOI: 10.1159/000516216 -
MBio Apr 2018Enterococci are important human commensals and significant opportunistic pathogens. Biofilm-related enterococcal infections, such as endocarditis, urinary tract...
Enterococci are important human commensals and significant opportunistic pathogens. Biofilm-related enterococcal infections, such as endocarditis, urinary tract infections, wound and surgical site infections, and medical device-associated infections, often become chronic upon the formation of biofilm. The biofilm matrix establishes properties that distinguish this state from free-living bacterial cells and increase tolerance to antimicrobial interventions. The metabolic versatility of the enterococci is reflected in the diversity and complexity of environments and communities in which they thrive. Understanding metabolic factors governing colonization and persistence in different host niches can reveal factors influencing the transition to biofilm pathogenicity. Here, we report a form of iron-dependent metabolism for where, in the absence of heme, extracellular electron transfer (EET) and increased ATP production augment biofilm growth. We observe alterations in biofilm matrix depth and composition during iron-augmented biofilm growth. We show that the gene encoding l-lactate dehydrogenase is required for iron-augmented energy production and biofilm formation and promotes EET. Bacterial metabolic versatility can often influence the outcome of host-pathogen interactions, yet causes of metabolic shifts are difficult to resolve. The bacterial biofilm matrix provides the structural and functional support that distinguishes this state from free-living bacterial cells. Here, we show that the biofilm matrix can immobilize iron, providing access to this growth-promoting resource which is otherwise inaccessible in the planktonic state. Our data show that in the absence of heme, l-lactate dehydrogenase promotes EET and uses matrix-associated iron to carry out EET. Therefore, the presence of iron within the biofilm matrix leads to enhanced biofilm growth.
Topics: Biofilms; Electron Transport; Energy Metabolism; Enterococcus faecalis; Iron; L-Lactate Dehydrogenase
PubMed: 29636430
DOI: 10.1128/mBio.00626-17 -
International Journal of Molecular... Nov 2022Enterococcus belongs to a group of microorganisms known as lactic acid bacteria (LAB), which constitute a broad heterogeneous group of generally food-grade...
Enterococcus belongs to a group of microorganisms known as lactic acid bacteria (LAB), which constitute a broad heterogeneous group of generally food-grade microorganisms historically used in food preservation. Enterococci live as commensals of the gastrointestinal tract of warm-blooded animals, although they also are present in food of animal origin (milk, cheese, fermented sausages), vegetables, and plant materials because of their ability to survive heat treatments and adverse environmental conditions. The biotechnological traits of enterococci can be applied in the food industry; however, the emergence of enterococci as a cause of nosocomial infections makes their food status uncertain. Recent advances in high-throughput sequencing allow the subtyping of bacterial pathogens, but it cannot reflect the temporal dynamics and functional activities of microbiomes or bacterial isolates. Moreover, genetic analysis is based on sequence homologies, inferring functions from databases. Here, we used an end-to-end proteomic workflow to rapidly characterize two bacteriocin-producing Enterococcus faecium (Efm) strains. The proteome analysis was performed with liquid chromatography coupled to a trapped ion mobility spectrometry-time-of-flight mass spectrometry instrument (TimsTOF) for high-throughput and high-resolution characterization of bacterial proteins. Thus, we identified almost half of the proteins predicted in the bacterial genomes (>1100 unique proteins per isolate), including quantifying proteins conferring resistance to antibiotics, heavy metals, virulence factors, and bacteriocins. The obtained proteomes were annotated according to function, resulting in 22 complete KEGG metabolic pathway modules for both strains. The workflow used here successfully characterized these bacterial isolates and showed great promise for determining and optimizing the bioengineering and biotechnology properties of other LAB strains in the food industry.
Topics: Animals; Enterococcus faecium; Bacteriocins; Proteomics; Enterococcus; Cheese
PubMed: 36430310
DOI: 10.3390/ijms232213830 -
Emerging Infectious Diseases 1998Enterococci, leading causes of nosocomial bacteremia, surgical wound infection, and urinary tract infection, are becoming resistant to many and sometimes all standard... (Review)
Review
Enterococci, leading causes of nosocomial bacteremia, surgical wound infection, and urinary tract infection, are becoming resistant to many and sometimes all standard therapies. New rapid surveillance methods are highlighting the importance of examining enterococcal isolates at the species level. Most enterococcal infections are caused by Enterococcus faecalis, which are more likely to express traits related to overt virulence but--for the moment--also more likely to retain sensitivity to at least one effective antibiotic. The remaining infections are mostly caused by E. faecium, a species virtually devoid of known overt pathogenic traits but more likely to be resistant to even antibiotics of last resort. Effective control of multiple-drug resistant enterococci will require 1) better understanding of the interaction between enterococci, the hospital environment, and humans, 2) prudent antibiotic use, 3) better contact isolation in hospitals and other patient care environments, and 4) improved surveillance. Equally important is renewed vigor in the search for additional drugs, accompanied by the evolution of new therapeutic paradigms less vulnerable to the cycle of drug introduction and drug resistance.
Topics: Anti-Bacterial Agents; Drug Resistance, Microbial; Drug Resistance, Multiple; Enterococcus; Enterococcus faecalis; Enterococcus faecium; Gram-Positive Bacterial Infections; Humans; Microbial Sensitivity Tests; Virulence
PubMed: 9621194
DOI: 10.3201/eid0402.980211 -
Microbiology Spectrum Dec 2022It was recently proposed that Enterococcus faecium colonizing the human gut (previous clade B) actually corresponds to Enterococcus lactis. Our goals were to develop a...
It was recently proposed that Enterococcus faecium colonizing the human gut (previous clade B) actually corresponds to Enterococcus lactis. Our goals were to develop a PCR assay to rapidly differentiate these species and to discuss the main phenotypic and genotypic differences from a clinical perspective. The pan-genome of 512 genomes of E. faecium and E. lactis strains was analyzed to assess diversity in genes between the two species. Sequences were aligned to find the best candidate gene for designing species-specific primers, and their accuracy was tested with a collection of 382 enterococci. E. lactis isolates from clinical origins were further characterized by whole-genome sequencing (Illumina). Pan-genome analysis resulted in 12 gene variants, with gene (rhomboid protease) being selected as the candidate for species differentiation. The nucleotide sequence of diverged by 90 to 92% between sets, which allowed species identification through PCR with 100% specificity and no cross-reactivity. E. lactis strains were greatly pan-susceptible and not host specific. Hospital E. lactis isolates were susceptible to clinically relevant antibiotics, lacked infection-associated virulence markers, and were associated with patients presenting risk factors for enhanced bacterial translocation. Here, we propose a PCR-based assay using for easy routine differentiation between E. faecium and E. lactis that could be implemented in different public health contexts. We further suggest that E. lactis, a dominant human gut species, can cross the gut barrier in severely ill, immunodeficient, and surgical patients. Knowing that bacterial translocation may be a sepsis promoter, the relevance of infections caused by E. lactis strains, even if they are pan-susceptible, should be explored. Enterococcus faecium is a WHO priority pathogen that causes severe and hard-to-treat human infections. It was recently proposed that E. faecium colonizing the human gut (previous clade B) actually corresponds to Enterococcus lactis; therefore, some of the human infections occurring globally are being misidentified. In this work, we developed a PCR-based rapid identification method for the differentiation of E. faecium and E. lactis and discussed the main phenotypic and genotypic differences of these species from a clinical perspective. We identified the gene as the best candidate, based on the phylogenomic analysis of 512 published pan-genomes, and validated the PCR assay with a comprehensive collection of 382 enterococci obtained from different sources. Further detailed analysis of clinical E. lactis strains showed that they are highly susceptible to antibiotics and lack the typical virulence markers of E. faecium but are able to cause severe human infections in immunosuppressed patients, possibly in part due to gut barrier translocation.
Topics: Humans; Anti-Bacterial Agents; Enterococcus faecium; Genome, Bacterial; Gram-Positive Bacterial Infections; Polymerase Chain Reaction; Enterococcus
PubMed: 36453910
DOI: 10.1128/spectrum.03268-22 -
Mikrobiyoloji Bulteni Apr 2017Enterococci have recently become important due to their increased isolation rates in community-based and nosocomial infections and resistance to many antibiotics,...
Enterococci have recently become important due to their increased isolation rates in community-based and nosocomial infections and resistance to many antibiotics, including glycopeptides. The aim of this study was to evaluate the antimicrobial susceptible patterns and virulence factors of various clinical specimens; urine (n= 149), blood (n= 38), wound (n= 17), stool (n= 13), and other (n= 12) with a total of 229 enterococci including 138 E.faecalis and 91 E.faecium isolates. Aggregation factor (AF), enterococcus surface protein (esp), cytolysins and gelatinase encoding genes (asa1, esp, cylM, cylBcyl A, cylll, cylls, gelE, respectively) were investigated by molecular methods. Haemolysin production and gelatinase were studied phenotypically. A total of 30 isolates, 29 of E.faecium and one of E.faecalis isolates were resistant to vancomycin. High-level gentamicin and high-level streptomycin resistance in E.faecalis were 40.7% and 63.7% however, they were 47.1% and 55.8% in E.faecalis isolates. All strains were susceptible to linezolid. Ampicillin, penicillin and vancomycin resistance in E.faecium isolates were found to be higher than E.faecalis isolates (p= 0.001, p= 0.008 and p< 0.001). Asa1 (p< 0.001), cylll (p= 0.002) and cylls (p< 0.001) as well as gelatinase activity in isolates of E.faecalis were significantly higher than the isolates of E.faecium (p< 0.001). The most common virulence genes in our study were asa1 gene (45%), cyLs gene (33.2%) and esp gene (32.3%). Ciprofloxacin resistance in cylLL and cyLs gene positive isolates of E.faecalis were significantly higher compared to isolates that do not contain these genes (p= 0.035 and p= 0.047). Likewise, haemolysin producing E.faecium isolates were significantly more resistant to vancomycin compared to isolates that do not produce hemolysin (p< 0.001). When the virulence factors of vancomycin resistant and susceptible isolates were compared, the esp gene level in VRE E.faecium isolates was found to be 24.1%, while no esp gene was found in VRE E.faecalis isolates. The existence of asa1was negative in both VRE E.faecium and VRE E.faecalis isolates. The activity of hemolysin was found 42.3% for E.faecalis and 19.3% for E.faecium. In vancomycin-sensitive enterococcus (VSE) species, esp gene activity was 35.1% for E.faecalis, 29.4% for E.faecium, asa1 gene activity was 60.8% for E.faecalis and 47.1% for E.faecium, hemolysin activity was 52.8% for E.faecalis and 23.5% for E.faecium. In our study, it was found that VSE isolates have more virulence genes than VRE isolates. It should be kept in mind that VRE can causeinfections which are difficult-to-treat especially in hospitalized patients and VSE have significant virulence factors that can cause severe infections.
Topics: Anti-Bacterial Agents; Bacteremia; Bacteriuria; Drug Resistance, Bacterial; Enterococcus faecalis; Enterococcus faecium; Feces; Gram-Positive Bacterial Infections; Humans; Vancomycin Resistance; Virulence Factors; Wound Infection
PubMed: 28566074
DOI: 10.5578/mb.53992