scholarly journals IDENTIFICATION OF BACTERIA CAUSING NECROTIC PULP WITH 16S rRNA GENE POLYMERASE CHAIN REACTION AND ANTIBIOTIC RESISTANCE TESTING AT THE DENTAL HOSPITAL IN SEKELOA, BANDUNG, INDONESIA

2017 ◽  
Vol 10 (6) ◽  
pp. 284
Author(s):  
Tina Rostinawati ◽  
Sulistiyaningsih Hadisoebroto ◽  
Yoppi Iskandar ◽  
Prasetyo Hadi Nugroho ◽  
Audria Amanda Tara
Keyword(s):  
Antibiotic Resistance ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Inhibition Zone ◽  
Resistance Testing ◽  
Rrna Gene ◽  
Bacterial Dna ◽  
Study Results ◽  
Dental Hospital ◽  
Polymerase Chain

Objective: This study aims to identify the bacteria that cause necrotic pulp in teeth of dental patients and test resistance of the bacteria found to antibiotics. Methods: Bacteria were taken with paper points that were inserted into the root canal of patients at the Dental Hospital of the Faculty of Dentistry, Universitas Padjadjaran, Sekeloa, Bandung. Bacteria were cultured and Gram-stained. Bacterial DNA was isolated to be identified by polymerase chain reactions 16S rRNA method against known sequences of bacterial DNA. Then, identified bacteria were tested for antibiotic resistance to tetracycline (30 μg), clindamycin (2 μg), amoxicillin (10 μg), and ampicillin (10 μg). Results: The 16s rRNA gene fragment of the main bacterium found was in 98% homology with 16S rRNA gene database in http://blast.ncbi.nlm. nih.gov, i.e., Pseudomonas aeruginosa and Acinetobacter schindleri. The results from inhibition zone of each antibiotic were 20.12 mm, 8.97 mm, 8.12 mm, and 8.03 mm for tetracycline, clindamycin, amoxicillin, and ampicillin, respectively, to P. aeruginosa. While inhibition zone of tetracycline to A. schindleri was 37.7 mm. Conclusion: Based on the study results, P. aeruginosa from patients with necrotic pulp samples were resistant to clindamycin, amoxicillin, ampicillin, and decreased activity to tetracycline. While Acinetobacter schindleri was still sensitive to tetracycline.

scholarly journals Molecular characterization of ancient algal mats from the McMurdo Dry Valleys, Antarctica

2011 ◽  
Vol 24 (2) ◽  
pp. 139-146 ◽  
Author(s):  
Doug E. Antibus ◽  
Laura G. Leff ◽  
Brenda L. Hall ◽  
Jenny L. Baeseman ◽  
Christopher B. Blackwood
Keyword(s):  
Polymerase Chain Reaction ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Rrna Gene ◽  
Dry Valleys ◽  
Chain Reaction ◽  
Bacterial Dna ◽  
Algal Mats ◽  
Polymerase Chain ◽  
Bacterial 16S Rrna Gene

AbstractThe McMurdo Dry Valleys possess a cold and dry climate which favours biomolecular preservation, and present the possibility for preservation of biological materials over long timescales. We examined patterns of bacterial DNA abundance and diversity in algal mats from 8–26 539 years of age. Bacterial DNA abundance was inferred from extractable DNA quantity and quantitative polymerase chain reaction targeting the bacterial 16S rRNA gene. Because damage to bacterial DNA could limit its availability for polymerase chain reaction, the efficacy of DNA repair by a commercially available kit was also examined. Polymerase chain reaction amplicons of the bacterial 16S rRNA gene were obtained from seven of eight samples. Bulk DNA abundance and bacterial 16S rRNA gene copy number of template DNA declined with increasing sample age consistent with expectations of accumulation of DNA damage in ancient materials. Clone libraries revealed age related patterns of abundance for some bacterial phylogenetic groups. For example, Firmicutes and several other lineages were abundant in ancient samples, but Cyanobacteria were absent. This points to a biased persistence of bacterial lineages that change over time since photosynthesis was active.

scholarly journals Detection of Prosthetic Hip Infection at Revision Arthroplasty by Immunofluorescence Microscopy and PCR Amplification of the Bacterial 16S rRNA Gene

1999 ◽  
Vol 37 (10) ◽  
pp. 3281-3290 ◽  
Author(s):  
Michael M. Tunney ◽  
Sheila Patrick ◽  
Martin D. Curran ◽  
Gordon Ramage ◽  
Donna Hanna ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Immunofluorescence Microscopy ◽  
Joint Infection ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Bacterial Dna ◽  
Prosthetic Joint ◽  
Culture Positive ◽  
Bacterial 16S Rrna Gene

In this study the detection rates of bacterial infection of hip prostheses by culture and nonculture methods were compared for 120 patients with total hip revision surgery. By use of strict anaerobic bacteriological practice during the processing of samples and without enrichment, the incidence of infection by culture of material dislodged from retrieved prostheses after ultrasonication (sonicate) was 22%. Bacteria were observed by immunofluorescence microscopy in 63% of sonicate samples with a monoclonal antibody specific forPropionibacterium acnes and polyclonal antiserum specific for Staphylococcus spp. The bacteria were present either as single cells or in aggregates of up to 300 bacterial cells. These aggregates were not observed without sonication to dislodge the biofilm. Bacteria were observed in all of the culture-positive samples, and in some cases in which only one type of bacterium was identified by culture, both coccoid and coryneform bacteria were observed by immunofluorescence microscopy. Bacteria from skin-flake contamination were readily distinguishable from infecting bacteria by immunofluorescence microscopy. Examination of skin scrapings did not reveal large aggregates of bacteria but did reveal skin cells. These were not observed in the sonicates. Bacterial DNA was detected in 72% of sonicate samples by PCR amplification of a region of the bacterial 16S rRNA gene with universal primers. All of the culture-positive samples were also positive for bacterial DNA. Evidence of high-level infiltration either of neutrophils or of lymphocytes or macrophages into associated tissue was observed in 73% of patients. Our results indicate that the incidence of prosthetic joint infection is grossly underestimated by current culture detection methods. It is therefore imperative that current clinical practice with regard to the detection and subsequent treatment of prosthetic joint infection be reassessed in the light of these results.

scholarly journals Microbiota in Exhaled Breath Condensate and the Lung

2017 ◽  
Vol 83 (12) ◽  
Author(s):  
Laura Glendinning ◽  
Steven Wright ◽  
Peter Tennant ◽  
Andrew C. Gill ◽  
David Collie ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Exhaled Breath Condensate ◽  
Operational Taxonomic Unit ◽  
Rrna Gene ◽  
Exhaled Breath ◽  
Bacterial Dna ◽  
Less Invasive ◽  
Colistimethate Sodium ◽  
Breath Condensate

ABSTRACT The lung microbiota is commonly sampled using relatively invasive bronchoscopic procedures. Exhaled breath condensate (EBC) collection potentially offers a less invasive alternative for lung microbiota sampling. We compared lung microbiota samples retrieved by protected specimen brushings (PSB) and exhaled breath condensate collection. We also sought to assess whether aerosolized antibiotic treatment would influence the lung microbiota and whether this change could be detected in EBC. EBC was collected from 6 conscious sheep and then from the same anesthetized sheep during mechanical ventilation. Following the latter EBC collection, PSB samples were collected from separate sites within each sheep lung. On the subsequent day, each sheep was then treated with nebulized colistimethate sodium. Two days after nebulization, EBC and PSB samples were again collected. Bacterial DNA was quantified using 16S rRNA gene quantitative PCR. The V2-V3 region of the 16S rRNA gene was amplified by PCR and sequenced using Illumina MiSeq. Quality control and operational taxonomic unit (OTU) clustering were performed with mothur. The EBC samples contained significantly less bacterial DNA than the PSB samples. The EBC samples from anesthetized animals clustered separately by their bacterial community compositions in comparison to the PSB samples, and 37 bacterial OTUs were identified as differentially abundant between the two sample types. Despite only low concentrations of colistin being detected in bronchoalveolar lavage fluid, PSB samples were found to differ by their bacterial compositions before and after colistimethate sodium treatment. Our findings indicate that microbiota in EBC samples and PSB samples are not equivalent. IMPORTANCE Sampling of the lung microbiota usually necessitates performing bronchoscopic procedures that involve a hospital visit for human participants and the use of trained staff. The inconvenience and perceived discomfort of participating in this kind of research may deter healthy volunteers and may not be a safe option for patients with advanced lung disease. This study set out to evaluate a less invasive method for collecting lung microbiota samples by comparing samples taken via protected specimen brushings (PSB) to those taken via exhaled breath condensate (EBC) collection. We found that there was less bacterial DNA in EBC samples compared with that in PSB samples and that there were differences between the bacterial communities in the two sample types. We conclude that while EBC and PSB samples do not produce equivalent microbiota samples, the study of the EBC microbiota may still be of interest.

PCR Analysis of Nasal Polyps, Chronic Sinusitis, and Hypertrophied Turbinates for DNA Encoding Bacterial 16S rRNA

2002 ◽  
Vol 16 (3) ◽  
pp. 169-173 ◽  
Author(s):  
Gerald A Bucholtz ◽  
Sherry A. Salzman ◽  
Fernando B. Bersalona ◽  
Timothy R. Boyle ◽  
Victor S. Ejercito ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Nasal Polyps ◽  
Chronic Sinusitis ◽  
Consensus Sequence ◽  
Rrna Gene ◽  
Dna Encoding ◽  
Bacterial Dna ◽  
Two Samples ◽  
The 16S Rrna Gene

Background Nasal polyps are considered to result from chronic inflammation, but the initial or persisting stimulus for the inflammation is not known. A variety of bacteria and fungi have been cultured from nasal polyps, but ∼35% have sterile cultures. Previously, Mycoplasma pneumoniae–specific DNA was detected in human nasal polyps using polymerase chain reaction (PCR) techniques, suggesting M. pneumoniae as a causative agent in the etiology of nasal polyps. Methods In this study, we tested for the presence of bacterial DNA in nasal polyps resected from 40 patients, in nasal mucosa membrane from 9 patients undergoing turbinectomy for hypertrophy, and in sinus mucosa membrane from 6 patients undergoing endoscopic surgery for chronic sinusitis. Tissue DNA was extracted and analyzed by PCR using M. pneumoniae specific primers for DNA that encode the 16S rRNA gene in 41 specimens (31 polyps, 6 turbinates, and 4 sinus), and by consensus sequence-based PCR using broad range primers for most eubacterial DNA encoding the 16S rRNA gene in 38 specimens (26 polyps, 7 turbinates, and 5 sinuses). Results Only two samples were positive for bacterial DNA encoding 16S rRNA: Streptococcus sp. DNA was isolated from one polyp specimen and Pseudomonas aeruginosa DNA was isolated in one maxillary sinusitis specimen. No evidence of M. pneumoniae–specific DNA encoding 16S rRNA was found in any of the tissues. Conclusions This study suggests that chronic bacterial infection is not a major component of nasal polyp etiology.

scholarly journals Effects of Antibiotics on the Bacterial Community, Metabolic Functions and Antibiotic Resistance Genes in Mariculture Sediments during Enrichment Culturing

2020 ◽  
Vol 8 (8) ◽  
pp. 604
Author(s):  
Meng-Qi Ye ◽  
Guan-Jun Chen ◽  
Zong-Jun Du
Keyword(s):  
Antibiotic Resistance ◽  
Bacterial Community ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
High Throughput ◽  
Antibiotic Resistance Genes ◽  
Control Group ◽  
Rrna Gene ◽  
Metabolic Functions ◽  
Culture Dependent

The effect of antibiotics on the diversity and functioning of indigenous microorganisms in the environment has attracted much attention. In this study, effects of exposure to six different antibiotics on the bacterial community, metabolic functions and antibiotic resistance genes (ARGs) in marine sediments during enrichment culturing were investigated. Classical culture-dependent method and high-throughput 16S rRNA gene sequencing method were both applied. In the culture-dependent analysis, the obtained 1549 isolates belonged to four phyla (Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria) and 155 genera. Proteobacteria and Firmicutes were the dominant phyla. The diversity and abundance of obtained bacteria after antibiotic processing exhibited different degrees of decrease. Enrichment culturing for different time could also affect the bacterial community composition. Some genera of bacteria were not isolated in the control group, but they could be isolated in the antibiotic-treated groups. In high-throughput 16S rRNA gene amplicon sequencing analyses, all the effective reads were clustered into 2822 OTUs at 97% similarity cutoff; they were annotated to 49 phyla, 103 class, 220 orders, 347 families, 624 genera and 1122 species. An alpha diversity analysis indicated that the community diversity and richness decreased under antibiotic exposure. The changes at the genus level were much more obvious. Only 48 genera of 129 genera were shared by all the samples. A total of 29 genera which were not detected in the initial control sample could be detected in at least one antibiotic-treated group. SIMPER analysis showed that OTU2543 and OTU1450 were the most common taxa to the dissimilarity of bacterial community between antibiotic-treated groups and the control group. OTU2034 and OUT2543 were the most contributive taxa to dissimilarity of groups incubating for different time. Metabolism was the predominant bacterial function. A total of 30 ARGs were detected in the samples. This study mainly focused on the changes of microbiota under the selective pressure of antibiotics for different time and the results demonstrated that the antibiotic could affect the bacterial diversity and richness in the marine ecosystem.

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