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MBio Jul 2014Bacterial DNA and live bacteria have been detected in human urine in the absence of clinical infection, challenging the prevailing dogma that urine is normally sterile....
Bacterial DNA and live bacteria have been detected in human urine in the absence of clinical infection, challenging the prevailing dogma that urine is normally sterile. Urgency urinary incontinence (UUI) is a poorly understood urinary condition characterized by symptoms that overlap urinary infection, including urinary urgency and increased frequency with urinary incontinence. The recent discovery of the urinary microbiome warrants investigation into whether bacteria contribute to UUI. In this study, we used 16S rRNA gene sequencing to classify bacterial DNA and expanded quantitative urine culture (EQUC) techniques to isolate live bacteria in urine collected by using a transurethral catheter from women with UUI and, in comparison, a cohort without UUI. For these cohorts, we demonstrated that the UUI and non-UUI urinary microbiomes differ by group based on both sequence and culture evidences. Compared to the non-UUI microbiome, sequencing experiments revealed that the UUI microbiome was composed of increased Gardnerella and decreased Lactobacillus. Nine genera (Actinobaculum, Actinomyces, Aerococcus, Arthrobacter, Corynebacterium, Gardnerella, Oligella, Staphylococcus, and Streptococcus) were more frequently cultured from the UUI cohort. Although Lactobacillus was isolated from both cohorts, distinctions existed at the species level, with Lactobacillus gasseri detected more frequently in the UUI cohort and Lactobacillus crispatus most frequently detected in controls. Combined, these data suggest that potentially important differences exist in the urinary microbiomes of women with and without UUI, which have strong implications in prevention, diagnosis, or treatment of UUI. Importance: New evidence indicates that the human urinary tract contains microbial communities; however, the role of these communities in urinary health remains to be elucidated. Urgency urinary incontinence (UUI) is a highly prevalent yet poorly understood urinary condition characterized by urgency, frequency, and urinary incontinence. Given the significant overlap of UUI symptoms with those of urinary tract infections, it is possible that UUI may have a microbial component. We compared the urinary microbiomes of women affected by UUI to those of a comparison group without UUI, using both high-throughput sequencing and extended culture techniques. We identified statistically significant differences in the frequency and abundance of bacteria present. These differences suggest a potential role for the urinary microbiome in female urinary health.
Topics: Actinomyces; Aerococcus; Aged; Arthrobacter; Corynebacterium; Female; Gardnerella; Humans; Lactobacillus; Microbiota; Middle Aged; RNA, Ribosomal, 16S; Staphylococcus; Streptococcus; Urinary Incontinence; Urinary Tract
PubMed: 25006228
DOI: 10.1128/mBio.01283-14 -
Emerging Infectious Diseases Jul 2015
Topics: Aged; Alcaligenaceae; Bacteremia; Female; Gram-Negative Bacterial Infections; Humans; United States
PubMed: 26079071
DOI: 10.3201/eid2107.150242 -
Atencion Primaria Nov 2001
Topics: Aged; Anti-Bacterial Agents; Female; Follow-Up Studies; Fosfomycin; Gram-Negative Bacterial Infections; Humans; Moraxella; Time Factors; Urinary Tract Infections; Urine
PubMed: 11747779
DOI: 10.1016/s0212-6567(01)70466-0 -
Zhongguo Dang Dai Er Ke Za Zhi =... Jul 2019To study the structural features of intestinal flora in preterm rats with cognitive impairment and the association of the change in intestinal flora with cognitive...
OBJECTIVE
To study the structural features of intestinal flora in preterm rats with cognitive impairment and the association of the change in intestinal flora with cognitive impairment in preterm rats.
METHODS
Sprague-Dawley rats at 16-17 days of gestation were intraperitoneally injected with lipopolysaccharide for two consecutive days to establish a model of cognitive impairment, and the rats treated with intraperitoneally injected phosphate-buffered saline were established as the control group. Cesarean section was performed on day 21 of gestation, and preterm rats were randomly assigned to healthy maternal rats for feeding. The place navigation test in the Morris water maze was used to evaluate cognition on day 30 after birth. According to the result, the preterm rats were divided into cognitive impairment group with 21 rats and normal control group with 10 rats. Hematoxylin and eosin staining was used to observe pathological changes of the hippocampus, and fecal samples were collected for 16S rRNA sequencing and analysis. A principal component analysis (PCA) was performed for intestinal flora.
RESULTS
Compared with the normal control group, the cognitive impairment group showed degeneration and necrosis of a large number of neurons in the hippocampus. Compared with the normal control group, the cognitive impairment group had significant reductions in the abundance and diversity of intestinal flora (P<0.05), with a significant increase in the abundance of Proteobacteria at the phylum level (P<0.05), as well as significant reductions in the abundance of Prevotella and Lactobacillus and significant increases in the abundance of Staphylococcaceae and Oligella at the order, family, and genus levels (P<0.05). PCA showed a significant difference in the composition of intestinal flora between the two groups.
CONCLUSIONS
There is a significant change in the structure of intestinal flora in preterm rats with cognitive impairment, which provides a basis for the treatment and intervention of microecological changes due to cognitive impairment after preterm birth.
Topics: Animals; Cesarean Section; Cognitive Dysfunction; Female; Gastrointestinal Microbiome; Pregnancy; RNA, Ribosomal, 16S; Rats; Rats, Sprague-Dawley
PubMed: 31315772
DOI: 10.7499/j.issn.1008-8830.2019.07.016 -
Antimicrobial Agents and Chemotherapy Jul 2005Acinetobacter spp. are emerging as opportunistic hospital pathogens that demonstrate resistance to many classes of antibiotics. In a metropolitan hospital in Cleveland,...
Acinetobacter spp. are emerging as opportunistic hospital pathogens that demonstrate resistance to many classes of antibiotics. In a metropolitan hospital in Cleveland, a clinical isolate of Acinetobacter baumannii that tested resistant to cefepime and ceftazidime (MIC = 32 microg/ml) was identified. Herein, we sought to determine the molecular basis for the extended-spectrum-cephalosporin resistance. Using analytical isoelectric focusing, a beta-lactamase with a pI of > or = 9.2 was detected. PCR amplification with specific A. baumannii cephalosporinase primers yielded a 1,152-bp product which, when sequenced, identified a novel 383-amino-acid class C enzyme. Expressed in Escherichia coli DH10B, this beta-lactamase demonstrated greater resistance against ceftazidime and cefotaxime than cefepime (4.0 microg/ml versus 0.06 microg/ml). The kinetic characteristics of this beta-lactamase were similar to other cephalosporinases found in Acinetobacter spp. In addition, this cephalosporinase was inhibited by meropenem, imipenem, ertapenem, and sulopenem (K(i) < 40 microM). The amino acid compositions of this novel enzyme and other class C beta-lactamases thus far described for A. baumannii, Acinetobacter genomic species 3, and Oligella urethralis in Europe and South Africa suggest that this cephalosporinase defines a unique family of class C enzymes. We propose a uniform designation for this family of cephalosporinases (Acinetobacter-derived cephalosporinases [ADC]) found in Acinetobacter spp. and identify this enzyme as ADC-7 beta-lactamase. The coalescence of Acinetobacter ampC beta-lactamases into a single common ancestor and the substantial phylogenetic distance separating them from other ampC genes support the logical value of developing a system of nomenclature for these Acinetobacter cephalosporinase genes.
Topics: Acinetobacter baumannii; Alleles; Amino Acid Sequence; Anti-Bacterial Agents; Bacterial Proteins; Base Sequence; Cephalosporin Resistance; Cephalosporinase; Genetic Variation; Humans; Microbial Sensitivity Tests; Molecular Sequence Data; Phylogeny; Sequence Analysis, DNA; beta-Lactamases; beta-Lactams
PubMed: 15980372
DOI: 10.1128/AAC.49.7.2941-2948.2005 -
The Journal of Veterinary Medical... Dec 1999In order to establish a rapid diagnostic method for contagious equine metritis (CEM), we developed and evaluated a polymerase chain reaction (PCR) test. Species-specific...
In order to establish a rapid diagnostic method for contagious equine metritis (CEM), we developed and evaluated a polymerase chain reaction (PCR) test. Species-specific PCR primer sets were derived from the DNA sequence of a cloned DNA fragment of Taylorella equigenitalis that did not hybridize with the genome of a taxomonically related species, Oligella urethralis. Single step PCR with primer set P1-N2 and two-step semi-nested PCR with primer sets P1-N2 and P2-N2 detected as low as 100 and 10 CFU of the bacteria, respectively. Single-step PCR detected T. equigenitalis from genital swabs of experimentally infected mares with sensitivity comparable to that of bacterial isolation. Furthermore, two-step PCR was more sensitive than the culture method. Upon examination of field samples, 12 out of 3,123 samples were positive by single-step PCR while only 2 were positive by bacterial culture. The 12 PCR-positive samples originated from 5 mares, of which 3 animals were considered to be carriers based on previous bacteriologic and serologic diagnoses for CEM. The PCR test described in this study would provide a specific and highly sensitive tool for the rapid diagnosis of CEM.
Topics: Animals; Base Sequence; Cervix Uteri; Clitoris; DNA Primers; DNA, Bacterial; Electrophoresis, Agar Gel; Endometritis; Female; Gene Library; Gram-Negative Bacterial Infections; Horse Diseases; Horses; Male; Molecular Sequence Data; Nucleic Acid Hybridization; Penis; Polymerase Chain Reaction; Sensitivity and Specificity; Sequence Analysis, DNA; Taylorella equigenitalis
PubMed: 10651048
DOI: 10.1292/jvms.61.1287 -
Infectious Diseases of Poverty Nov 2016Previously rare A2ML1 variants were identified to confer otitis media susceptibility in an indigenous Filipino community and in otitis-prone US children. The goal of...
BACKGROUND
Previously rare A2ML1 variants were identified to confer otitis media susceptibility in an indigenous Filipino community and in otitis-prone US children. The goal of this study is to describe differences in the middle ear microbiome between carriers and non-carriers of an A2ML1 duplication variant that increases risk for chronic otitis media among indigenous Filipinos with poor health care access.
METHODS
Ear swabs were obtained from 16 indigenous Filipino individuals with chronic otitis media, of whom 11 carry the A2ML1 duplication variant. Ear swabs were submitted for 16S rRNA gene sequencing.
RESULTS
Genotype-based differences in microbial richness, structure, and composition were identified, but were not statistically significant. Taxonomic analysis revealed that the relative abundance of the phyla Fusobacteria and Bacteroidetes, and genus Fusobacterium were nominally increased in carriers compared to non-carriers, but were non-significant after correction for multiple testing. We also detected rare bacteria including Oligella that was reported only once in the middle ear.
CONCLUSIONS
These findings suggest that A2ML1-related otitis media susceptibility may be mediated by changes in the middle ear microbiome. Knowledge of middle ear microbial profiles according to genetic background can be potentially useful for therapeutic and prophylactic interventions for otitis media and can guide public health interventions towards decreasing otitis media prevalence within the indigenous Filipino community.
Topics: Adolescent; Child; Child, Preschool; DNA, Bacterial; Ear, Middle; Female; Genes, Duplicate; Humans; Male; Microbiota; Otitis Media; Philippines; Population Groups; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Young Adult; alpha-Macroglobulins
PubMed: 27799062
DOI: 10.1186/s40249-016-0189-7 -
Journal of Clinical Microbiology Jun 1990A pseudomonad was isolated from the pleural fluid and pulmonary decortication tissue of a 5-year-old child with chronic granulomatous disease. Although the isolate was... (Comparative Study)
Comparative Study
A pseudomonad was isolated from the pleural fluid and pulmonary decortication tissue of a 5-year-old child with chronic granulomatous disease. Although the isolate was phenotypically similar to Pseudomonas cepacia, its biochemical profile was more similar to that of Pseudomonas pickettii biovar 2. Its slow growth rate, ability to hydrolyze urea rapidly, and lateral and polar flagellar pattern were suggestive of Oligella ureolytica (formerly CDC group IVe). The cellular fatty acid composition was similar to that of P. cepacia and Pseudomonas gladioli, except for the presence of dodecanoic acid. Numerical analysis of the fatty acid data supported the interrelatedness of the isolate with other species of the pseudomallei group (rRNA homology group II) of Pseudomonas. The organism described in this report is an addition to the growing list of catalase-positive organisms which can potentially cause severe morbidity in patients with chronic granulomatous disease.
Topics: Child, Preschool; Emphysema; Fatty Acids; Granulomatous Disease, Chronic; Humans; Male; Pneumonia; Pseudomonas; Pseudomonas Infections; RNA, Ribosomal; Sequence Homology, Nucleic Acid
PubMed: 2380349
DOI: 10.1128/jcm.28.6.1120-1124.1990