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Research in Microbiology 2021Ralstonia pickettii are ubiquitous in water environments. Members of this species are frequently, but not always, resistant to both gentamicin and arsenite. Gentamicin...
Ralstonia pickettii are ubiquitous in water environments. Members of this species are frequently, but not always, resistant to both gentamicin and arsenite. Gentamicin and arsenite co-resistance and the putative molecular mechanisms were investigated. A group of 37 R. pickettii strains isolated from drinking water and hospital wastewater were characterized for gentamicin and arsenite resistance phenotypes, the number and size of plasmids, and screened for genetic elements associated with arsenite tolerance, Integrative and Conjugative Elements (ICEs), among other. The genomes of three representative strains were compared. Most gentamicin resistant (GR) isolates (32/33) were resistant to arsenite, and harbored ICE- and ars operon-related genes. These genetic elements were not detected in any of the five arsenite susceptible strains, regardless of the GR (n = 1) or gentamicin susceptibility (GS) (n = 4) phenotype. The comparison of the genomes of two GR (one resistant and one susceptible to arsenite) and one GS strains suggested that these phenotypes correspond to three phylogroups, distinguished by presence of some genes only in GR isolates, in addition to point mutations in functional genes. The presence of ICEs and ars operon-related genes suggest that arsenite resistance might have been acquired by GR lineages.
Topics: Anti-Bacterial Agents; Arsenites; Conjugation, Genetic; Drinking Water; Drug Resistance, Multiple, Bacterial; Genome, Bacterial; Gentamicins; Humans; Microbial Sensitivity Tests; Ralstonia pickettii; Wastewater
PubMed: 33197514
DOI: 10.1016/j.resmic.2020.11.001 -
Journal of Infection in Developing... Apr 2022Ralstonia pickettii infections are rare and may be mistaken for other bacteria. This study aims to report a hospital outbreak of R. pickettii at a tertiary hospital,...
INTRODUCTION
Ralstonia pickettii infections are rare and may be mistaken for other bacteria. This study aims to report a hospital outbreak of R. pickettii at a tertiary hospital, which was initially misidentified as Ralstonia insidiosa, along with its clinical consequences.
METHODOLOGY
A bacteraemia outbreak occurred between August 14 and October 4, 2019, infecting 22 patients admitted to diverse intensive care units. All isolates were identified with the use of the automated VITEK 2 Compact system and were then subjected to a microbial identification system, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Bacterial identification and genomic DNA typing was made using pulsed-field gel electrophoresis. Investigation covered all potential sources of the outbreak.
RESULTS
An index patient and five additional patients developed fever while receiving care. Blood cultures of these patients yielded R. insidiosa by the VITEK 2 Compact system. Culture isolates were then submitted to a reference centre for confirmation by the MALDI-TOF MS system, where the bacterium turned out to be R. pickettii. No pathogen was isolated in the commercial products except for three samples of unopened sterile distilled water. Despite its discontinuation, 16 new cases were identified, in which blood cultures grew R. pickettii by the MALDI-TOF MS system. Attempts to uncover the source of the outbreak failed. Clinical manifestation was confined to fever in all the patients.
CONCLUSIONS
During this outbreak, R. pickettii infections ran a relatively mild course without clinical deterioration or mortality, possibly due to low virulence.
Topics: Bacteremia; Disease Outbreaks; Electrophoresis, Gel, Pulsed-Field; Humans; Ralstonia pickettii; Sepsis; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 35544634
DOI: 10.3855/jidc.15159 -
Journal of Personalized Medicine Apr 2021The microbiota has been reported to be correlated with carcinogenesis and cancer progression. However, its involvement in the pathology of mesothelioma remains unknown....
The microbiota has been reported to be correlated with carcinogenesis and cancer progression. However, its involvement in the pathology of mesothelioma remains unknown. In this study, we aimed to identify mesothelioma-specific microbiota using resected or biopsied mesothelioma samples. Eight mesothelioma tissue samples were analyzed via polymerase chain reaction (PCR) amplification and 16S rRNA gene sequencing. The operational taxonomic units (OTUs) of the effective tags were analyzed in order to determine the taxon composition of each sample. For the three patients who underwent extra pleural pneumonectomy, normal peripheral lung tissues adjacent to the tumor were also included, and the same analysis was performed. In total, 61 OTUs were identified in the tumor and lung tissues, which were classified into 36 species. and were identified as abundant species in almost all tumor and lung samples. and were found to comprise mesothelioma-specific microbiota involved in tumor progression; thus, they could serve as targets for the prevention of mesothelioma.
PubMed: 33919754
DOI: 10.3390/jpm11040297 -
Journal of Microbiology, Immunology,... Feb 2022Ralstonia pickettii is an opportunistic waterborne microbe which can survive in many kinds of solutions. Contamination of these solutions may result as outbreaks, which...
BACKGROUND
Ralstonia pickettii is an opportunistic waterborne microbe which can survive in many kinds of solutions. Contamination of these solutions may result as outbreaks, which can be mortal for immuncompromised patients. Herein we report an outbreak of R. pickettii related to contaminated saline infusion in our center.
METHODS
This study was conducted in Ankara Pediatric City Hospital. An outbreak occured in Pediatric Hematology and Oncology Unit between August 28, 2019 and September 13, 2019. When the outbreak occured, infection control team began an investigation. Environmental samples were collected in order to find the source of the outbreak.
RESULTS
A total of 11 patients with catheter related blood stream infection caused by R. pickettii who were diagnosed with leukemia were affected. None of the patients infected with R. pickettii died during the outbreak. A total of seventy environmental samples were cultured with the purpose of finding the source of outbreak. R. pickettii grew in normal saline solution culture and all isolates had the same clone of R. pickettii. The outbreak lasted two weeks and was controlled by stopping the usage and sending back the saline solutions belonging to the same manufacturing batch.
CONCLUSIONS
We reported an outbreak of R. pickettii BSIs in highly immunocompromised patients due to contaminated intravascular solution, which was rapidly controlled by infection control measures. Vigilant surveillance by hospital infection control teams and prompt investigation to identify the source of nosocomial infections are crucial to stop an outbreak.
Topics: Child; Cross Infection; Disease Outbreaks; Humans; Leukemia; Ralstonia pickettii; Sepsis
PubMed: 33461864
DOI: 10.1016/j.jmii.2020.12.004 -
Medicine Jul 2019By 2030, the annual number of combined total hip and knee arthroplasty is estimated to reach 3.5 to 4 million in the US alone. In the context of a constant increase of... (Observational Study)
Observational Study
By 2030, the annual number of combined total hip and knee arthroplasty is estimated to reach 3.5 to 4 million in the US alone. In the context of a constant increase of the number of primary and revision total hip and knee arthroplasty, an increased risk of complication is expected. Prosthetic joint infections (PJIs) represent major cause of healthcare expenditure and morbidity. PJI still remain the most common and feared arthroplasty complication. A rapid and correct diagnosis of infection is decisive for a correct therapeutical management. In this setting, the Academic Emergency Hospital Sibiu adopted and implemented, with the beginning of September 2016, a new strategy for the diagnosis of PJIs strategy that uses sonication and beacon-based fluorescent in situ hybridization (bbFISH) technology.Until November 2017, 40 patients (40 retrieved implants) were enrolled in the study. Sonication fluid (SF) was collected after sonication of the implants, and samples were harvested on aerobic and anaerobic culture media. A bbFISH was used as a rapid method of bacteria detection.16 patients were diagnosed with PJIs (all 16 patients presented a positive culture of the SF). Comparing bbFISH with culture, 11 samples tested true-positive. As the kit doesn't contain probes for Pseudomonas fluorescens or Ralstonia pickettii, 4 strains of R pickettii and 1 strain of P fluorescens that was associated with Staphylococcus epidermidis were not detected.Bacteria culture of SF remains the gold standard. bbFISH holds promise to be a diagnostic tool for rapid identifying of PJIs. The bbFISH assay needs to be optimized for the detection of bacterial strains that are relevant for the PJIs field.
Topics: Aged; Aged, 80 and over; Arthroplasty, Replacement, Hip; Arthroplasty, Replacement, Knee; Bacterial Infections; Bacteriological Techniques; Connective Tissue; Female; Humans; In Situ Hybridization, Fluorescence; Male; Middle Aged; Prosthesis-Related Infections; RNA, Bacterial; RNA, Ribosomal, 16S; Sonication; Synovial Fluid
PubMed: 31335719
DOI: 10.1097/MD.0000000000016501 -
Antonie Van Leeuwenhoek Oct 2021To improve understanding of the role of Ralstonia in cystic fibrosis (CF), whole genomes of 18 strains from clinical samples were sequenced using Illumina technology....
To improve understanding of the role of Ralstonia in cystic fibrosis (CF), whole genomes of 18 strains from clinical samples were sequenced using Illumina technology. Sequences were analysed by core genome Multi-Locus Sequence Typing, Average Nucleotide Identity based on BLAST (ANIb), RAST annotation, and by ResFinder. Phylogenetic analysis was performed for the 16S rRNA gene, and the OXA-22 and OXA-60 ß-lactamase families. The minimal inhibitory concentrations (MICs) were determined using broth microdilution. ANIb data for the 18 isolates and 54 strains from GenBank, supported by phylogenetic analysis, showed that 8 groups of clusters (A-H), as well as subgroups that should be considered as species or subspecies. Groups A-C contain strains previously identified as Ralstonia solanacearum and Ralstonia pseudosolanacearum. We propose that group A is a novel species. Group B and C are Ralstonia syzygii, Ralstonia solanacearum, respectively. Group D is composed of Ralstonia mannitolilytica and Group E of Ralstonia pickettii. Group F and G should be considered novel species. Group H strains belong to R. insidiosa. OXA-22 and OXA-60 family ß-lactamases were encoded by all strains. Co-trimoxazole generally showed high activity with low MICs (≤1 mg/l) as did ciprofloxacin (≤0.12 mg/l). MICs against the other antibiotics were more variable, but generally high. RAST annotation revealed limited differences between the strains, and virulence factors were not identified. The taxonomy of the genus Ralstonia is in need of revision, but sequencing additional isolates is needed. Antibiotic resistance levels are high. Annotation did not identify potential virulence factors.
Topics: Humans; Multilocus Sequence Typing; Phylogeny; RNA, Ribosomal, 16S; Ralstonia
PubMed: 34463860
DOI: 10.1007/s10482-021-01637-0 -
Journal of Microbiology and... Sep 2019DDT is a hydrophobic organic pollutant, which can be bio-accumulated in nature and have adverse consequences on the physical condition of humans and animals. This study...
DDT is a hydrophobic organic pollutant, which can be bio-accumulated in nature and have adverse consequences on the physical condition of humans and animals. This study investigated the relationship between the white-rot fungus and biosurfactantproducing bacterium associated with the degradation of DDT. The effects of on fungal development were examined using in vitro confrontation assay on a potato dextrose agar (PDA) medium. culture was added to the culture at 1, 3, 5, 7, and 10 ml (1 ml ≈ 1.44 × 10 CFU). After 7 d incubation, about 43% of the initial DDT (12.5 μM) was degraded by the culture only. The augmentation of 7 ml of culture revealed a more highly optimized synergism with DDT degradation being approximately 78% and the ratio of optimization 1.06. According to the confrontational assay, promoted the growth of towards the bacterial colony, with no direct contact between the bacterial cells and mycelium (0.71 cm/day). DDD (1,1-dichloro-2,2-bis(4- chlorophenyl) ethane), DDE (1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene), and DDMU (1- chloro-2,2-bis(4-chlorophenyl) ethylene) were identified as metabolic products, indicating that the could enhance the DDT biodegradation by .
Topics: Biodegradation, Environmental; Coculture Techniques; DDT; Insecticides; Pleurotus; Ralstonia pickettii
PubMed: 31474097
DOI: 10.4014/jmb.1906.06030 -
Microbiology Spectrum Dec 2022Growing evidence indicates an association between gut dysbiosis and coronary artery disease (CAD). However, the underlying mechanisms relevant to stable CAD (SCAD)...
Growing evidence indicates an association between gut dysbiosis and coronary artery disease (CAD). However, the underlying mechanisms relevant to stable CAD (SCAD) pathogenesis, based on microbe-host metabolism interactions, are poorly explored. Here, we constructed a quasi-paired cohort based on the metabolic background of metagenomic samples by the propensity score matching (PSM) principle. Compared to healthy controls (HCs), gut microbiome disturbances were observed in SCAD patients, accompanied by differences in serum metabolome, mainly including elevated acylcarnitine and decreased unsaturated fatty acids in SCAD patients, which implicated the reduced cardiac fatty acid oxidation. Moreover, we identified Ralstonia pickettii as the core strain responsible for impaired microbial homeostasis in SCAD patientsm and may be partly responsible for the decrease of host unsaturated fatty acid levels. These findings highlight the importance of unsaturated fatty acids, R. pickettii, and their interaction in the pathogenesis of SCAD. Stable coronary artery disease (SCAD) is an early stage of CAD development. It is important to understand the pathogenesis of SCAD and find out the possible prevention and control targets for delaying the progression of CAD. We observed reduced levels of unsaturated fatty acids (USFAs) in SCAD patients. However, the reduced USFAs may be related to Ralstonia Pickettii, which was the core strain responsible for the impaired gut microbial function in SCAD patients, and further affected the host's cardiovascular health by altering amino acids, vitamin B metabolism, and LPS biosynthesis. These findings not only emphasized the importance of USFAs for cardiovascular health, but also R. Pickettii for maintaining microbial function homeostasis. More importantly, our study revealed, for the first time, that enriched R. Pickettii might be responsible for the reduced USFAs in SCAD patients, which adds new evidence on the role of altered gut microbiota for SCAD formation.
Topics: Humans; Coronary Artery Disease; Gastrointestinal Microbiome; Metabolome; Metagenomics; Lipid Metabolism
PubMed: 36354350
DOI: 10.1128/spectrum.02467-22 -
Nan Fang Yi Ke Da Xue Xue Bao = Journal... Dec 2022To develop a method for rapid detection of in water for pharmaceutical purpose using PCR-nucleic acid test strips.
OBJECTIVE
To develop a method for rapid detection of in water for pharmaceutical purpose using PCR-nucleic acid test strips.
METHODS
The genomic DNA of was extracted by boiling method. A pair of specific primers targeting the 16S rDNA with FITC and biotin labeling of the 5' ends was designed and cloned into competent DH5α cells. The nucleic acid test strips were assembled, and the workload of streptavidin labeled with colloidal gold and antibody concentration in the reaction system was optimized. After verification of the reaction mechanism and assessment of the test sensitivity, specificity and stability, the test strip was used for detecting 7 known strains of detected in pharmaceutical water, and an evolutionary tree was constructed to analyze the source of contamination.
RESULTS
The genomic DNA extracted by boiling method had a purity between 1.8 and 2.0, and the PCR products showed a 100% similarity of with 16S rDNA registered in GenBank. Using the colloidal gold amplification principle, in every 100 μL colloidal gold solution, 3.5 μL streptavidin was added; the detection line on nitrocellulose membrane was 2.0 mg·mL anti FITC antibody, and the quality control line was 1.2 mg · mL biotinylated BSA, and they generate a red band after binding with positive amplification product. Specificity test of the assembled test strip yielded consistent result with agarose gel electrophoresis without cross reaction with , , , or . Sensitivity test of the strip showed a lower detection limit for DNA concentration of 10 ng/μL, with a sensitivity 1000 times that of agarose gel electrophoresis. The test strip still had good performance after storage for 3, 6, 9 and 12 months.
CONCLUSION
We successfully developed a PCR-nucleic acid test strip for convenient and cost-effective detection of with good specificity and sensitivity and low cost to facilitate daily monitoring of pharmaceutical water contaminations.
Topics: DNA, Ribosomal; Escherichia coli; Gold Colloid; Nucleic Acids; Polymerase Chain Reaction; Ralstonia pickettii; Reagent Strips; Streptavidin; Water
PubMed: 36651256
DOI: 10.12122/j.issn.1673-4254.2022.12.16 -
Frontiers in Cellular and Infection... 2023Bladder cancer (BCa) is the most common malignancy of the urinary tract which can be divided into non-muscle-invasive bladder cancer (NMIBC) and muscle-invasive bladder...
BACKGROUND
Bladder cancer (BCa) is the most common malignancy of the urinary tract which can be divided into non-muscle-invasive bladder cancer (NMIBC) and muscle-invasive bladder cancer (MIBC), and their microbial differences are not fully understood. This study was conducted by performing 2bRAD sequencing for Microbiome (2bRAD-M) on NMIBC and MIBC tissue samples to investigate the microbiota differences between NMIBC and MIBC individuals.
METHODS
A total of 22 patients with BCa, including 7 NMIBC and 15 MIBC, were recruited. Tumor tissues were surgically removed as samples and DNA was extracted. Type IIB restriction endonucleases were used to enzymatically cleave the microbial genome for each microbe's tag and map it to species-specific 2bRAD markers to enable qualitative and quantitative studies of microbes between MIBC and NMIBC tissues.
RESULTS
A total of 527 species were detected. The microbial diversity of NMIBC tissues was significantly higher than that of MIBC tissues. Microbial composition of the two tumor tissues was similar, where Ralstonia_sp000620465 was the most dominant species. 4 species (Acinetobacter_guillouiae, Anoxybacillus_A_rupiensis, Brevibacillus_agri and Staphylococcus_lugdunensis) were enriched in NMIBC, while Ralstonia_mannitolilytica, Ralstonia_pickettii, and Ralstonia_sp000620465 were overrepresented in MIBC. 252 discriminatory character taxa were also revealed by linear discriminant analysis effect sizea (LEfSe). Species importance point plots identified Ralstonia_sp000620465, Cutibacterium_acnes and Ralstonia_pickettii as the three most important species between the two groups. Meanwhile, functional annotation analysis showed 3011 different COGs and 344 related signaling pathways between MIBC and NMIBC microbiome.
CONCLUSION
This first 2bRAD-M microbiome study on MIBC and NMIBC tissues revealed significant differences in the microbial environment between the two groups, which implies a potential association between tumor microbial dysbiosis and BCa, and provides a possible target and basis for subsequent studies on the mechanisms of BCa development and progression.
Topics: Humans; Urinary Bladder; Urinary Bladder Neoplasms; Neoplasm Invasiveness
PubMed: 37351184
DOI: 10.3389/fcimb.2023.1182322