Identification Of Specific Bacteria Research Articles

Variations in oral microbiome and its predictive functions between tumorous and healthy individuals.

Introduction. The oral cavity is one of the largest reservoirs of microorganisms and many pathogenic bacteria have been shown to be associated with the aetiology of oral cancers.Gap Statement. Owing to the complexity of oral microbial communities and their unclear relationship with oral cancer, identification of specific bacteria which contribute to oral cancer is a key imperative.Aim. To compare and investigate the variations in the composition of the bacterial microbiome and its functions between patients with oral tumorous lesions and healthy subjects.Methodology. Twenty-seven samples from individuals with oral tumours (five oral benign tumours and 22 oral squamous cell carcinomas) and 15 samples from healthy subjects were collected. Genomic DNA was extracted and the V3-V5 region of the 16S rRNA gene was sequenced. Subsequently, bioinformatic assessment was conducted using QIIME2, PICRUSt and linear discriminant analysis effect size analyses (LEfSe).Results. The oral microbiota was composed mainly of the phyla Proteobacteria (31.76 %, 35.00 %), Bacteroidetes (30.13 %, 25.13 %) and Firmicutes (23.92 %, 17.07 %) in tumorous and healthy individuals, respectively. Neisseria, Prevotella, Fusobacterium, Streptococcus, Capnocytophaga, Veillonella, Haemophilus, Prevotella, Porphyromonas and Leptotrichia were the most abundant genera. Alpha diversity in the tumour group was significantly greater than that in the healthy group (P<0.05). Differential analysis of microbes between groups demonstrated a significantly higher number of Neisseria, Veillonella, Streptococcus, Leptotrichia, Lautropia, Sphingopyxis, Sphingobium, Tannerella, Actinomyces and Rothia in healthy controls compared with the tumour group. However, the genera Treponema, Micrococcus, Pseudomonas, Janthinobacterium, Parvimos, Loktanella, Staphylococcus, Acinetobacter, Catonella, Aggregatibacter and Propionibacterium were significantly higher in the tumour group. Pathways related to cancers, cell motility, environmental adaptation, metabolism and signal transduction were enhanced in the tumour group, while functions associated with immune system diseases, replication, repair and translation were significantly enhanced in the healthy group.Conclusion. Variations in the oral microbiota and its functions showed a correlation with oral tumours. The tumour group showed an increased abundance of some multi-drug-resistant and periodontitis-related pathogens. The significantly altered microbiotas may serve as potential biomarkers or inform combination therapy for oral tumours.

The Oral Bacterial Microbiome of Interdental Surfaces in Adolescents According to Carious Risk.

Adolescence is closely associated with a high risk of caries. The identification of specific bacteria in an oral microniche, the interdental space of the molars, according to carious risk can facilitate the prediction of future caries and the anticipation of the progression or stabilization of caries in adolescents. A cross-sectional clinical study according to the bacteriological criteria of interdental healthy adolescents and carious risk factors—low and high—using a real-time polymerase chain reaction technique was conducted. The presence of 26 oral pathogens from the interdental microbiota of 50 adolescents aged 15 to 17 years were qualitatively and quantitatively analyzed. Bacteria known to be cariogenic (Bifidobacterium dentium, Lactobacillus spp., Rothia dentocariosa, Streptococcus cristatus, Streptococcus mutans, Streptococcus salivarius, Streptococcus sobrinus, and Streptococcus wiggsiae) did not present differences in abundance according to carious risk. Periodontal bacteria from the red complex are positively correlated with carious risk. However, only 3 bacteria—S. sobrinus, E corrodens and T. forsythia—presented a significant increase in the highest group. Estimating the risk of caries associated with bacterial factors in interdental sites of molars in adolescents contributes to the better definition of carious risk status, periodicity and intensity of diagnostic, prevention and restorative services.

417 Identification of Specific Bacteria in Jejunal Aspiration Is an Important Determinant of Small Intestinal Bacterial Overgrowth (SIBO)

417 Identification of Specific Bacteria in Jejunal Aspiration Is an Important Determinant of Small Intestinal Bacterial Overgrowth (SIBO)

MALDI-TOF identification of the human Gut microbiome in people with and without diarrhea in Senegal.

BackgroundIn Africa, there are several problems with the specific identification of bacteria. Recently, MALDI-TOF mass spectrometry has become a powerful tool for the routine microbial identification in many clinical laboratories.Methodology/Principal FindingsThis study was conducted using feces from 347 individuals (162 with diarrhea and 185 without diarrhea) sampled in health centers in Dakar, Senegal. Feces were transported from Dakar to Marseille, France, where they were cultured using different culture conditions. The isolated colonies were identified using MALDI-TOF. If a colony was unidentified, 16S rRNA sequencing was performed. Overall, 2,753 isolates were tested, allowing for the identification of 189 bacteria from 5 phyla, including 2 previously unknown species, 11 species not previously reported in the human gut, 10 species not previously reported in humans, and 3 fungi. 2,718 bacterial isolates (98.8%) out of 2,750 yielded an accurate identification using mass spectrometry, as did the 3 Candida albicans isolates. Thirty-two bacterial isolates not identified by MALDI-TOF (1.2%) were identified by sequencing, allowing for the identification of 2 new species. The number of bacterial species per fecal sample was significantly higher among patients without diarrhea (8.6±3) than in those with diarrhea (7.3±3.4; P = 0.0003). A modification of the gut microbiota was observed between the two groups. In individuals with diarrhea, major commensal bacterial species such as E. coli were significantly decreased (85% versus 64%), as were several Enterococcus spp. (E. faecium and E. casseliflavus) and anaerobes, such as Bacteroides spp. (B. uniformis and B. vulgatus) and Clostridium spp. (C. bifermentans, C. orbiscindens, C. perfringens, and C. symbosium). Conversely, several Bacillus spp. (B. licheniformis, B. mojavensis, and B. pumilus) were significantly more frequent among patients with diarrhea.Conclusions/SignificanceMALDI-TOF is a potentially powerful tool for routine bacterial identification in Africa, allowing for a quick identification of bacterial species.

Label-Free and Selective Bacteria Detection Using a Film with Transferred Bacterial Configuration

Specific identification of bacteria has been achieved through precisely transferred bacterial structure on the surface of overoxidized polypyrrole (OPPy) film. The recognition of target bacteria was successfully carried out in real time using OPPy film in combination with dielectrophoresis. The unique combination of both techniques made the specific detection of template bacilli possible at concentrations as low as 10(3) CFU/mL within 3 min, without any bacterial pretreatment. The observation of the movement of bacteria by using a fluorescent microscope revealed that living bacteria were being trapped vertically in the cavity created in the OPPy film. Further, the bacterial cavities had high selectivity and were able to discriminate particular target bacteria, Pseudomonas aeruginosa, out of bacterial mixtures containing Acinetobacter calcoaceticus, Escherichia coli, and Serratia marcescens, which are known to have a similar shape. This simple method can be used for a wide variety of applications in which rapid bacterial detection is required, such as food safety risk assessment, clinical point-of-care testing, and continuous environmental monitoring.

Central nervous system marginal zone B-cell lymphoma associated with Chlamydophila psittaci infection

Extranodal marginal zone B-cell lymphomas are linked to bacterial infections that vary according to the anatomical site. The occurrence of these lymphomas in the central nervous system is a very rare event, and the identification of specific bacteria in this setting has not been previously addressed. Herein, we report for the first time a case of primary central nervous system marginal zone B-cell lymphoma involving the choroid plexus associated with Chlamydophila psittaci infection. No concomitant ocular involvement was detected. C psittaci was identified with 3 independent methods, and through immunohistochemistry, it was visualized in the cytoplasm of monocytes/macrophages present within lymphomatous tissues. This observation points toward the opportunity to investigate the prevalence of C psittaci infection in central nervous system lymphomas, particularly in those with low-grade histologic features.

Detection and identification of specific bacteria in wound biofilms using peptide nucleic acid fluorescent in situ hybridization (PNA FISH)

Biofilms provide a reservoir of potentially infectious micro-organisms that are resistant to antimicrobial agents, and their importance in the failure of medical devices and chronic inflammatory conditions is increasingly being recognized. Particular research interest exists in the association of biofilms with wound infection and non-healing, i.e. chronic wounds. In this study, fluorescent in situ hybridization (FISH) was used in combination with confocal laser scanning microscopy (CLSM) to detect and characterize the spatial distribution of biofilm-forming bacteria which predominate within human chronic skin wounds (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus sp. and Micrococcus sp.). In vitro biofilms were prepared using a constant-depth film fermenter and a reconstituted human epidermis model. In vivo biofilms were also studied using biopsy samples from non-infected chronic venous leg ulcers. The specificity of peptide nucleic acid (PNA) probes for the target organisms was confirmed using mixed preparations of planktonic bacteria and multiplex PNA probing. Identification and location of individual bacterial species within multi-species biofilms demonstrated that P. aeruginosa was predominant. CLSM revealed clustering of individual species within mixed-species biofilms. FISH analysis of archive chronic wound biopsy sections showed bacterial presence and allowed bacterial load to be determined. The application of this standardized procedure makes available an assay for identification of single- or multi-species bacterial populations in tissue biopsies. The technique provides a reliable tool to study bacterial biofilm formation and offers an approach to assess targeted biofilm disruption strategies in vivo.

The Ibis T5000 Universal Biosensor: An Automated Platform for Pathogen Identification and Strain Typing

We describe a new approach to the sensitive and specific identification of bacteria, viruses, fungi, and protozoa based on broad-range PCR and high-performance mass spectrometry. The Ibis T5000 is based on technology developed for the Department of Defense known as T.I.G.E.R. (Triangulation Identification for the Genetic Evaluation of Risks) for pathogen surveillance. The technology uses mass spectrometry—derived base composition signatures obtained from PCR amplification of broadly conserved regions of the pathogen genomes to identify most organisms present in a sample. The process of sample analysis has been automated using a combination of commercially available and custom instrumentation. A software system known as T-Track manages the sample flow, signal analysis, and data interpretation and provides simplified result reports to the user. No specialized expertise is required to use the instrumentation. In addition to pathogen surveillance, the Ibis T5000 is being applied to reducing health care—associated infections (HAIs), emerging and pandemic disease surveillance, human forensics analysis, and pharmaceutical product and food safety, and will be used eventually in human infectious disease diagnosis. In this review, we describe the automated Ibis T5000 instrument and provide examples of how it is used in HAI control.

Identification and strain differentiation of Vibrio cholerae by using polyclonal antibodies against outer membrane proteins.

Cholera is caused only by O1 and O139 Vibrio cholerae strains. For diagnosis, 3 working days are needed for bacterial isolation from human feces and for biochemical characterization. Here we describe the purification of bacterial outer membrane proteins (OMP) from V. cholerae O1 Ogawa, O1 Inaba, and O139 strains, as well as the production of specific antisera and their use for fecal Vibrio antigen detection. Anti-OMP antisera showed very high reactivity and specificity by enzyme-linked immunosorbent assay (ELISA) and dot-ELISA. An inmunodiagnostic assay for V. cholerae detection was developed; this assay avoids preenrichment and costly equipment and can be used for epidemiological surveillance and clinical diagnosis of cases, considering that prompt and specific identification of bacteria is mandatory in cholera.

The specific and sensitive detection of bacterial pathogens within 4 h using bacteriophage amplification.

This paper describes a novel approach, termed the 'phage amplification assay', for the rapid detection and identification of specific bacteria. The technique is based on the phage lytic cycle with plaque formation as the assay end-point. It is highly sensitive, quantitative and gives results typically within 4 h. The assay comprises four main stages: (1) phage infection of target bacterium; (2) destruction of exogenous phage; (3) amplification of phage within infected host and (4) plaque formation from infected host with the aid of helper bacteria. A key component of this assay is a potent virucidal agent derived from natural plant extracts, pomegranate rind extract (PRE). In combination with ferrous sulphate PRE can bring about an 11 log-cycle reduction in phage titre within 3 min. This is achieved without any injury to the infected target bacteria. Subsequently, any resulting plaques are derived only from infected target organisms. Data are presented for a range of bacterial hosts including Pseudomonas aeruginosa, Salmonella typhimurium and Staphylococcus aureus. The detection limit for Ps. aeruginosa was 40 bacteria ml-1 in a time of 4 h and 600 bacteria m-1 for Salm. typhimurium. Application of the principles of this technology to other bacterial genera is discussed.

Detection of Rhizobium meliloti cells in field soil and nodules by polymerase chain reaction.

A genetically marked Rhizobium meliloti strain, R692, was prepared by insertion of a 1.7-kb DNA segment from Tn903 between the nifHDK and fixABC genes in the nod megaplasmid. This DNA was used as a marker, detectable by polymerase chain reaction (PCR), for the specific identification of bacteria in soil samples and alfalfa nodules. This detection technique was tested by applying different titres of the marked strain to field plots seeded with alfalfa. Samples of soil and nodules were assayed for the presence of the marker DNA fragment by PCR using primers specific to the marker sequence. The experiments revealed that the bacteria could be detected directly in soil containing about 10(3)-10(4) bacteria/g, but greater sensitivity was prevented by potent PCR inhibitors present in the samples. The titre of the bacteria in the soil decreased rapidly after inoculation, dropping about 10-fold per week. Tests of vertical location of the bacteria in soil cores showed that the bacteria were initially dispersed to a depth of 18 cm, and subsequently retained viability in the top 2-8 cm. As few as 10 marked R. meliloti per gram of soil resulted in its establishment at detectable levels in nodules. Application of about 10(4)-10(5) bacteria/g soil was sufficient to give the maximum number of nodules per plant and resulted in 70-90% occupancy by the marked strain. Limited movement of the inoculant was detected by analysis of nodules from plants adjacent to the sites where the bacteria were applied, probably by movement in water. The experiments demonstrated the advantages of PCR for the monitoring of marked microorganisms in the environment.

The presence of bacteria within the oral epithelium in periodontal disease. I. A scanning and transmission electron microscopic study.

The presence of bacteria within the gingival oral epithelium and adjacent connective tissue in cases of periodontitis and localized juvenile periodontitis have been described using scanning and transmission electron microscopy. The following bacterial morphotypes were identified: cocci, short rods, filaments and few spirochetes in periodontitis and mainly coccobacillary-shaped bacteria in localized juvenile periodontitis. Also Mycoplasma-like structures were identified in the localized juvenile periodontitis cases. Tunnel-like formations at different depths of the oral epithelium contained higher numbers of bacteria than those seen on the adjacent oral surface. Identification of specific bacteria in the oral epithelium may have important pathogenic and therapeutic implications.

Microbiologic Surveillance In Hospitals

To the Editor.— In response to a query regarding microbiologic sampling of the hospital environment, Carl W. Walter, MD, cited instances in which such studies have demonstrated sources of nosocomial infection (229: 578, 1974). Dr. Walter did not indicate specifically whether routine sampling is a worthwhile control measure. Except for identification of specific bacteria associated with individual outbreaks, no correlation has been established between levels of contamination and nosocomial infection. If infection-control committees placed primary emphasis on surveillance of infections and education of personnel, if sampling were inexpensive and were generally conducted, using standard methods, and if clinically significant ranges of contamination were known, routine sampling might be justified. Too often, what in fact occurs is subrogation of the responsibility of the staff and administration to the unstandardized collection of cultures, which is subsidized at apparently no cost by the laboratory. Until the true cost of conducting an effective program is