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At least 451 records · Page 25Linked to original sources
PubMed1993

Clonal differentiation of uropathogenic Escherichia coli isolates of serotype O6:K5 by fimbrial antigen typing and DNA long-range mapping techniques.

Escherichia coli isolates of serotype O6:K5 are the most common causative agents of cystitis and pyelonephritis in adults. To answer the question, as to whether strains of this particular serotype represent one special clonal group, out of a collection of 34 serotype O6:K5 isolates [Zingler et al. (1990) Zentralbl. Bakteriol Mikrobiol Hyg [A] 274:372-381] 15 strains were selected and analyzed in detail. The flagellar (H) antigen and the outer membrane protein (OMP) pattern were determined. Further serum resistance properties and the genetic presence and expression of other virulence factors, including hemolysin, aerobactin, P fimbriae, S/F1C fimbriae and type 1 fimbriae was evaluated. In addition the XbaI-macrorestriction pattern of ten representative isolates was elaborated and the fimbrial (F) antigen type of the P fimbriae was determined, to obtain the complete O:K:H:F pattern. These analyses could clearly show that the O6:K5 isolates do not represent one clonal group. The XbaI-macrorestriction profiles were heterogeneous and marked differences in the hybridization patterns, using virulence-associated gene probes in Southern hybridization of long-range-separated genomic DNA, were observed among the strains. However, some of strains showed similarities in the genomic profiles, arguing for clonal groupings among the O6:K5 isolates. Interestingly the strains grouped together exhibited the same fimbrial F type that many indicate a coincidence of this phenotypic trait with clonality.

Adhesins, Escherichia coli↗
PubMed1993

Molecular genotype analysis of Salmonella bovismorbificans.

Genotypes were analysed within 30 isolates of Salmonella bovismorbificans from human or bovine salmonellosis and from environmental or food sources. Three clonal evolutionary lines (chromosomal genotypes) were identified on the basis of restriction fragment length polymorphism at the 16S rRNA genes, total protein profiles, and the location and copy number of a Salmonella-specific DNA insertion sequence, IS200. The predominant type was found with and without a 90 kbp plasmid homologous to the spv BC genes of S. typhimurium, and seven plasmid profiles were observed in this chromosomal genotype. This is the first such virulence plasmid to be reported in S. bovismorbificans. One example was found of two other clones which differed substantially from the predominant clone and from each other in chromosomal genotype. These results show that three clonal lines share the antigen profile of S. bovismorbificans, and provide a subtyping scheme for this serovar.

Animals↗
PubMed2005

Comparison of Helicobacter pylori virulence gene expression in vitro and in the Rhesus macaque.

We used a quantitative real-time reverse transcriptase PCR assay to measure the transcript abundance of 46 known and putative Helicobacter pylori virulence genes, including 24 genes on the Cag pathogenicity island. The expression profile of H. pylori cells grown in vitro was also compared to expression in vivo after experimental infection of rhesus macaques. Transcript abundance in vitro (mid-log phase) ranged from about 0.004 (feoB and hpaA) to 20 (ureAB, napA, and cag25) copies/cell. Expression of most genes was repressed during the transition from logarithmic- to stationary-phase growth, but several well-characterized H. pylori virulence genes (katA, napA, vacA, and cagA) were induced. Comparison of results in the rhesus macaque with similar data from humans showed a strong correlation (r = 0.89). The relative in vivo expression in the rhesus monkey was highly correlated with in vitro expression during mid-log (r = 0.89)- and stationary (r = 0.88)-phase growth. Transcript abundance was on average three- to fourfold reduced in vivo compared to in vitro during mid-log phase. However, when compared to stationary phase, increased expression in vivo was observed for 6 of 7 genes on a contiguous portion of the pathogenicity island, several of which are thought to encode the H. pylori type IV structural pilus and its accessory proteins. These results suggest the possibility that some genes encoding the H. pylori type IV structural pilus and accessory proteins may form an operon that is induced during growth in vivo.

Animals↗
PubMed2004

Identification of Cryptococcus neoformans temperature-regulated genes with a genomic-DNA microarray.

The ability to survive and proliferate at 37 degrees C is an essential virulence attribute of pathogenic microorganisms. A partial-genome microarray was used to profile gene expression in the human-pathogenic fungus Cryptococcus neoformans during growth at 37 degrees C. Genes with orthologs involved in stress responses were induced during growth at 37 degrees C, suggesting that a conserved transcriptional program is used by C. neoformans to alter gene expression during stressful conditions. A gene encoding the transcription factor homolog Mga2 was induced at 37 degrees C and found to be important for high-temperature growth. Genes encoding fatty acid biosynthetic enzymes were identified as potential targets of Mga2, suggesting that membrane remodeling is an important component of adaptation to high growth temperatures. mga2Delta mutants were extremely sensitive to the ergosterol synthesis inhibitor fluconazole, indicating a coordination of the synthesis of membrane component precursors. Unexpectedly, genes involved in amino acid and pyrimidine biosynthesis were repressed at 37 degrees C, but components of these pathways were found to be required for high-temperature growth. Our findings demonstrate the utility of even partial-genome microarrays for delineating regulatory cascades that contribute to microbial pathogenesis.

Cryptococcus neoformans↗
PubMed2001

Molecular analysis of clinical isolates of Providencia alcalifaciens.

In an attempt to elucidate the virulence factors and the pathogenic mechanisms of Providencia alcalifaciens, 36 isolates identified in 1994-1995 in Recife city, Brazil were analysed by PCR to investigate the presence of DNA sequences homologous to virulence genes described in other invasive enterobacteria, as well as their ability to invade HeLa cells, their plasmid profiles and antibiotic resistance patterns. The genetic diversity of the isolates was also analysed by RAPD-PCR. No homologous sequences of virulence genes were observed with any of the P. alcalifaciens isolates studied. Ten isolates had no plasmid and 26 harboured one-to-five plasmids of 147-<6.9 kb. Invasion of HeLa cells was observed in only 10 isolates. No correlation between the plasmid content of the strains, their invasion of HeLa cells or their resistance to antimicrobial drugs could be established. The isolates could be distributed into 10 genotypic groups by RAPD-PCR. Considering the genotypic profile and ability to invade HeLa cells, 7 of the 10 invasive isolates belonged to the same genotypic group. The presence of invasive isolates in the same or a related genotypic group suggests the existence of a clonal lineage responsible for the invasiveness.

Anti-Bacterial Agents↗
PubMed2003

Apoptosis genes in human alveolar macrophages infected with virulent or attenuated Mycobacterium tuberculosis: a pivotal role for tumor necrosis factor.

Tumor necrosis factor (TNF)-alpha-dependent apoptosis of alveolar macrophages (AM) after infection with avirulent Mycobacterium tuberculosis (Mtb) results in bacillary death and the destruction of a growth niche for the pathogen. This response is minimized after infection with virulent strains of Mtb. To study the genetic control of Mtb-induced apoptosis, we used microarrays to interrogate the expression profile of infected human AM. Although we found variation in gene expression between different donors of AM, a set of genes were constant for each condition. A group of proapoptotic genes were downregulated after infection by virulent Mtb strain H37Rv, whereas infection with avirulent Mtb H37Ra led to a gene expression profile that would favor macrophage apoptosis. Neutralizing TNF in macrophage cultures infected with H37Ra changed the gene expression profile to one that resembled the profile of macrophages infected with H37Rv. These data reveal that apoptosis-related genes are regulated differently by virulent or attenuated Mtb strains, and are consistent with the hypothesis that virulent Mtb interfere with TNF death signaling. Given the importance of TNF in host defense against tuberculosis, the ability to repress the expression of genes activated by TNF may constitute a bacillary virulence mechanism.

Apoptosis↗
PubMed2007

FNR is a global regulator of virulence and anaerobic metabolism in Salmonella enterica serovar Typhimurium (ATCC 14028s).

Salmonella enterica serovar Typhimurium must successfully transition the broad fluctuations in oxygen concentrations encountered in the host. In Escherichia coli, FNR is one of the main regulatory proteins involved in O2 sensing. To assess the role of FNR in serovar Typhimurium, we constructed an isogenic fnr mutant in the virulent wild-type strain (ATCC 14028s) and compared their transcriptional profiles and pathogenicities in mice. Here, we report that, under anaerobic conditions, 311 genes (6.80% of the genome) are regulated directly or indirectly by FNR; of these, 87 genes (28%) are poorly characterized. Regulation by FNR in serovar Typhimurium is similar to, but distinct from, that in E. coli. Thus, genes/operons involved in aerobic metabolism, NO. detoxification, flagellar biosynthesis, motility, chemotaxis, and anaerobic carbon utilization are regulated by FNR in a fashion similar to that in E. coli. However, genes/operons existing in E. coli but regulated by FNR only in serovar Typhimurium include those coding for ethanolamine utilization, a universal stress protein, a ferritin-like protein, and a phosphotransacetylase. Interestingly, Salmonella-specific genes/operons regulated by FNR include numerous virulence genes within Salmonella pathogenicity island 1 (SPI-1), newly identified flagellar genes (mcpAC, cheV), and the virulence operon (srfABC). Furthermore, the role of FNR as a positive regulator of motility, flagellar biosynthesis, and pathogenesis was confirmed by showing that the mutant is nonmotile, lacks flagella, is attenuated in mice, and does not survive inside macrophages. The inability of the mutant to survive inside macrophages is likely due to its sensitivity to the reactive oxygen species generated by NADPH phagocyte oxidase.

Anaerobiosis↗
PubMed2005

[Molecular epidemilogy, antimicrobial and disinfectant resistance of Escherichia coli 0175:H7 in Shichuan].

OBJECTIVE: To study epidemiological characteristics of Escherichia coli O157:H7 isolated in Sichuan. METHODS: Multiplex PCR was used to analyze the slt1, slt2, eaeA, and hly gene. Plasmid profile and PFGE was utilized to investigate the homology among the strains. Drug and disinfectant resistance patterns of 67 stains were determined. RESULTS: The overall virulence gene prevalence was 62.7% (42/67); and the main virulence gene type was slt1 + slt2 + eaeA + hly. Of the 67 Escherichia coli O157: H7 strains, 5 plasmid profile and 6 PFGE types were acquired. 64.2% (43/67) Escherichia coli O157:H7 strains resisted to 7 antibiotics, namely, Ampicilin, Chloramphnicol, Tetracyline, Gentamicin, Cephlothin, SMZ, TMP-SMZ, and there were 23, 35, 34 strains resisted to Ampicilin, Tetracyline and SMZ respectively. The resistance types were different in Escherichia coli O157: H7 strains isolated from humans, animals, food, and environment. 80.6% (54/67) Escherichia coli 0157:H7 strains resisted to alcohol and quaternary ammonium, but all the stains were sensitive to chlorhexidiniacetas. CONCLUSION: The virulence gene prevalence rate was familiar of 67 Escherichia coli O157: H7 Strains isolated from Sichuan, but virulence gene type was not the same in different sources. Bacterial drug resistance was high. Bacterial resistance to common disinfectants was so high that the strains could be killed by big dosage disinfectants. There were some familiarity with drug resistance, plasmid profile and PFGE types of 67 Escherichia coli 0157:H7 strains isolated from different sources, which indicated homology of the strains.

Ampicillin↗
PubMed1999

In vivo genetic analysis of bacterial virulence.

In vitro assays contribute greatly to our understanding of bacterial pathogenesis, but they frequently cannot replicate the complex environment encountered by pathogens during infection. The information gained from such studies is therefore limited. In vivo models, on the other hand, can be difficult to use, and this has to some extent diminished the incentive to perform studies in living animals. However, several recently developed techniques permit in vivo examination of many genes simultaneously. Most of these methods fall into two broad classes: in vivo expression technology and signature-tagged mutagenesis. In vivo expression technology is a promoter-trap strategy designed to identify genes whose expression is induced in a specific environment, typically that encountered in a host. Signature-tagged mutagenesis uses comparative hybridization to isolate mutants unable to survive specified environmental conditions and has been used to identify genes critical for survival in the host. Both approaches have so far been used exclusively for investigating pathogen-host interactions, but they should be easily adaptable to the study of other processes.

Animals↗
PubMed2006

Comparison of the proteome profiles of Entamoeba histolytica and its close but non-pathogenic relative Entamoeba dispar.

The microaerophilic protist Entamoeba histolytica is a source of considerable morbidity and mortality in many countries of the world by acting as a causative agent of amoebic dysentery and liver abscess. Its close relative, Entamoeba dispar, also colonises the human colon but is non-pathogenic. In the present study, we wanted to assess if the close relatedness of these two organisms could be used to identify virulence factors in E. histolytica through a comparison of the protein expression profiles of the pathogenic and the non-pathogenic species. We applied two-dimensional polyacrylamide gel electrophoresis in order to identify proteins which are specifically expressed in E. histolytica and which therefore could constitute candidate proteins potentially involved in E. histolytica pathogenicity. The evaluation of overall protein expression profiles derived from whole cell extracts, however, revealed that, in spite of the close relatedness at the genetic level, the disparity of the proteomic profiles of these two Entamoeba species is far too great in order to pinpoint proteins whose expression might render E. histolytica pathogenic. Taken together, our study performed at the proteomic level clearly supports the notion of E. histolytica and E. dispar as two distinct species.

Animals↗
PubMed2006

Occurrence of virulence and antimicrobial resistance genes in Escherichia coli isolates from different aquatic ecosystems within the St. Clair River and Detroit River areas.

Although the number of Escherichia coli bacteria in surface waters can differ greatly between locations, relatively little is known about the distribution of E. coli pathotypes in surface waters used as sources for drinking or recreation. DNA microarray technology is a suitable tool for this type of study due to its ability to detect high numbers of virulence and antimicrobial resistance genes simultaneously. Pathotype, phylogenetic group, and antimicrobial resistance gene profiles were determined for 308 E. coli isolates from surface water samples collected from diverse aquatic ecosystems at six different sites in the St. Clair River and Detroit River areas. A higher frequency (48%) of E. coli isolates possessing virulence and antimicrobial resistance genes was observed in an urban site located downstream of wastewater effluent outfalls than in the other examined sites (average of 24%). Most E. coli pathotypes were extraintestinal pathogenic E. coli (ExPEC) pathotypes and belonged to phylogenetic groups B2 and D. The ExPEC pathotypes were found to occur across all aquatic ecosystems investigated, including riverine, estuarine, and offshore lake locations. The results of this environmental study using DNA microarrays highlight the widespread distribution of E. coli pathotypes in aquatic ecosystems and the potential public health threat of E. coli pathotypes originating from municipal wastewater sources.

Bacterial Proteins↗
PubMed2026

Molecular epidemiology and genomic characteristics of clinical Acinetobacter baumannii isolates from patients with hospital-acquired pneumonia in China, 2019-2020: a multicentre retrospective study.

BACKGROUND: Carbapenem-resistant Acinetobacter baumannii (CRAB) is a leading cause of hospital-acquired pneumonia (HAP) with high mortality. However, large-scale nationwide data of HAP-causing CRAB in China remain limited. METHODS: Here, we performed a nationwide multicentre retrospective study to characterise the molecular epidemiology and genomic features of 802 A. baumannii isolates from patients with HAP across 33 tertiary hospitals in China during 2019-2020. Antimicrobial susceptibility testing (AST), whole-genome sequencing (WGS), phylogenetic and comparative genomic analysis were used to investigate molecular epidemiology of HAP-causing CRAB strain. Clinical comparative analyses were carried out on data from 500 patients with HAP stratified by distinct antimicrobial susceptibility and genomic profiles, and a Galleria mellonella infection model was utilised for in vivo virulence assessment. FINDINGS: The overall carbapenem resistance rate of A. baumannii was 82.0% (658/802), with marked regional variations. CRAB exhibited high resistance to conventional agents but remained largely susceptible to polymyxin, tigecycline, cefiderocol and sulbactam-durlobactam. Among enrolled patients, CRAB infection was linked to substantially higher mortality (39.0% vs. 17.5%), and multivariate analysis confirmed ICU admission and advanced age as independent risk factors for patients with CRAB infection. Molecular typing revealed STPas2 (96.2%) as the absolutely predominant type; STOxf208, STOxf195, STOxf540, and STOxf369 were the most prevalent Oxford sequence types with obvious geographic stratification and divergent comorbidity profiles among corresponding patients. A total of 654 CRAB isolates harboured carbapenemase genes, with blaOXA-23 dominating at 98.8%. Genomic analysis revealed lineage-specific features: STOxf208 carried more virulence genes, while STOxf540 harboured a broader antimicrobial resistance genes (ARGs). The STOxf208 clone mainly belonged to KL2 (62.1%) and KL7 (36.8%) serotypes, with KL2 strains possessing richer ARGs and virulence factors, and in vivo virulence assays further validated that KL2 strains possessed higher pathogenicity than KL7 strains. INTERPRETATION: This study demonstrates the extremely high prevalence and clonal dominance of CRAB in Chinese patients with HAP, providing critical evidence for clinical treatment, antimicrobial stewardship, and targeted infection control. FUNDING: National Key Research and Development Program of China (2024YFE0106200), National Natural Science Foundation of China (U22A20338, 82502763, W2621007), Zhejiang Provincial Natural Science Foundation of China (LQN25H190006), Zhejiang Provincial Postdoctoral Science Foundation (ZJ2025058).

Acinetobacter baumannii↗
PubMed1994

O-serogrouping and surface components of Aeromonas hydrophila and Aeromonas jandaei pathogenic for eels.

The relationship between virulence, O-serogroup, and some cell-surface features (self-pelleting [SP] and precipitation after boiling [PAB], profile of lipopolysaccharides [LPSs] and outer membrane proteins [OMPs]) was investigated in strains of the pathogenic species Aeromonas hydrophila and A. jandaei isolated from eels. Virulent strains of A. hydrophila reacted mostly with O:19 antiserum, and those of A. jandaei reacted with O:4, O:11, O:15 and O:29 antisera (Guinée and Jansen system). Regarding the PAB and LPS profiles two groups could be distinguished; (i) five PAB+ strains of serotype O:19 that possessed a homogeneous O polysaccharide side chain and (ii) thirteen PAB- strains antigenically diverse that either exhibited a heterogeneous side chain or were side chain deficient. A major 50 kDa protein was only found in the PAB+ strains, whereas major OMPs detected in PAB- strains ranged from 33 to 45 kDa irrespective of the species. Epizootic eel isolates of A. hydrophila belong to serotype O:19 and share cell-surface features with the Aeromonas highly virulent for other hosts. In contrast, epizootic A. jandaei isolates were antigenically diverse. These findings reinforce the importance of an O-serotype as an epidemiological marker in motile Aeromonas strains pathogenic for eels.

Aeromonas↗
PubMed2000

[Study on mRNA differential display in Mycobacterium tuberculosis H37Rv and H37Ra].

OBJECTIVE: To study genetic differences between H37Rv and H37Ra by mRNA differential display, paving the way for obtaining virulence-related gene of Mycobacterium tuberculosis (MTB). METHODS: Total RNA was extracted from MTB H37Rv and H37Ra, mRNA differential display was conducted following reverse transcription, polymerase chain reaction, polyacrilamide gel electrophoresis and radiography. The differential bands were recovered, reamplified, sequenced, and then homologous query was done. RESULTS: 10 differential bands between H37Rv and H37Ra were recovered, 2 of which were homologous to H37Rv DNA after query from NCBI database of GenBank. CONCLUSIONS: mRNA differential display is an effective, sensitive method, can display differences between different strains intra- and extra-species and be used in searching Mycobacterium tuberculosis virulence-related gene. Obtained differential bands in this study may cover MTB virulence-related gene.

Animals↗
PubMed2026

Comprehensive profiling of antibiotic resistance genes and functional clusters of orthologous groups annotation of gut microbiota in Indonesian Kedu chickens.

Antibiotic resistance is a growing global health concern, with poultry systems acting as important reservoirs of antibiotic resistance genes (ARGs). However, resistome and functional profiles of indigenous chickens raised under traditional systems remain underexplored. This study aimed to characterize the antibiotic resistome, virulence factor genes, and metabolic potential of gut microbiota in Indonesian Kedu chickens using a shotgun metagenomic approach. Digesta samples from five gastrointestinal segments of 21 healthy adult chickens were analyzed through high-throughput sequencing. ARGs were identified using the Comprehensive Antibiotic Resistance Database (CARD) and Antibiotic Resistance Genes Databases (ARDB), while virulence factors and functional genes were annotated using Virulence Factor Database (VFDB), Clusters of Orthologous Groups (COG), and Carbohydrate-Active EnZymes (CAZy) databases. Results revealed a diverse resistome dominated by multidrug resistance and efflux pump mechanisms, with prominent genes associated with fluoroquinolone, tetracycline, &#x3b2;-lactam, and glycopeptide resistance. The detection of clinically relevant ARGs suggests that genetic determinants associated with antimicrobial resistance are present in the gut microbiota of traditionally raised Kedu chickens, although metagenomic data alone cannot determine whether these genes are actively expressed or confer phenotypic resistance. Virulence factor analysis showed functions related to adherence, immune evasion, iron acquisition, quorum sensing, and efflux activity, reflecting strong microbial adaptability. Functional profiling demonstrated enrichment in translation, carbohydrate and amino acid metabolism, genome maintenance, and cell envelope biogenesis. Additionally, CAZyme analysis indicated a high capacity for complex polysaccharide degradation, supporting efficient utilization of fiber-rich traditional diets. In conclusion, this study provides a comprehensive metagenomic overview of antibiotic resistance and functional potential in Kedu chicken gut microbiota, emphasizing the importance of incorporating indigenous poultry into antimicrobial resistance surveillance within a One Health framework.

Antibiotic resistance genes↗
PubMed2006

Prevalence of shiga toxin-producing Escherichia coli in a diarrheagenic Tunisian population, and the report of isolating STEC O157:H7 in Tunis.

In Mellassine (a major city in the state of Tunis) and Ben Arous state (south east of Tunis), a total of 212 stool samples were collected from children and adults (symptomatic and asymptomatic groups) between November 2001 and November 2004. Three hundred and twenty-seven E. coli strains were isolated and studied, to look for shiga toxin-producing Escherichia coli (STEC) strains, which were further analysed to investigate and determine clonal relationship among Tunisian STEC strains isolated from different sources (diarrheal cases and food products). They were analysed to characterize their serotypes, virulence genes by PCR, cytotoxic effect on Vero cell, plasmid profiles, and pulsed-field gel electrophoresis (PFGE) patterns. Eleven isolates (10 nontypeable, one O157:H7) carried stx gene and shared Stx restriction fragment length polymorphism (RFLP) patterns (stx1 ( + ), stx2 ( + )). Seven of these strains were isolated from acute diarrheal cases, and four were isolated from a control group (among which the only isolated STEC O157:H7). Two of the STEC strains harboured both eae and ehxA genes. Analysis of the cytotoxic effect on Vero cells showed that a correlation exists between carrying stx1 ( + ), stx2 ( + ) genes and cytotoxicity. Also a correlation was noticed between STEC strains recovered from different sources regarding plasmid profiles and PFGE patterns. All stool samples positive for STEC were nonbloody. None of the STEC-positive patients developed severe diseases. These data demonstrate that although STEC is not a major cause of acute diarrhea in Tunis, it should not be overlooked. Measures should be taken to improve the detection and isolation of STEC from acute diarrheal cases as well as carriers.

Adult↗
PubMed1995

Oral Candida albicans biotypes in Chinese patients with and without oral candidosis.

A total of 53 oral Candida albicans isolates from Chinese patients with clinically diagnosed oral candidosis (27 patients) or without overt signs and mycological manifestations of infection (26) were biotyped using two commercially available API micromethod kits and a boric acid-resistance test. There were no significant differences in the biotypes in health and disease, although the biotype A1R was present only in diseased individuals. The biotype A1S accounted for 21% of the total isolates, as in a number of other previous studies from the West. However, 14 of the 27 biotypes characterized were new biotypes that have not been described before. These preliminary data indicate that biotypic profile of C. albicans may bear no relation to the virulence of the isolates, and that diverse subtypes of the fungus are globally prevalent.

Boric Acids↗