RESUMEN
"Bacteroides denticanum" is an anaerobic, non-spore-forming, gram-negative bacterium with a rod morphology typical of canine, ovine, and macropod oral flora. There is only one report of bloodstream infection caused by "B. denticanum" from a dog bite in human. Here, we report a case with no history of animal contact who developed an abscess caused by "B. denticanum" around a pharyngo-esophageal anastomosis after undergoing balloon dilatation procedure for stenosis following laryngectomy. The patient was a 73-year-old man with laryngeal cancer, esophageal cancer, hyperuricemia, dyslipidemia, and hypertension with a 4-week history of cervical pain, sore throat, and fever. Computed tomography showed fluid collection on the posterior pharyngeal wall. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) identified Bacteroides pyogenes, Lactobacillus salivarius, and Streptococcus anginosus from abscess aspiration. 16S ribosomal RNA sequencing re-identified the Bacteroides species as "B. denticanum". T2-weighted magnetic resonance images showed a high signal intensity adjacent to the anterior vertebral body of C3-C7. The diagnosis was peripharyngeal esophageal anastomotic abscess and acute vertebral osteomyelitis caused by "B. denticanum", L. salivarius, and S. anginosus. The patient was treated with sulbactam ampicillin intravenously for 14 days and then switched to oral amoxicillin with clavulanic acid for 6 weeks. To our knowledge, this is the first report of a human infection caused by "B. denticanum" without a history of animal contact. Despite remarkable advancements facilitated by MALDI-TOF MS in microbiological diagnosis, the accurate identification of novel, emerging, or uncommon microorganisms and comprehending their pathogenicity, suitable therapy, and follow up necessitate sophisticated molecular approaches.
Asunto(s)
Coinfección , Ligilactobacillus salivarius , Osteomielitis , Masculino , Humanos , Animales , Perros , Ovinos/genética , Anciano , Streptococcus anginosus/genética , ARN Ribosómico , Absceso/diagnóstico , Absceso/tratamiento farmacológico , Bacteroides , ARN Ribosómico 16S/genética , Análisis de Secuencia de ARN , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Anastomosis Quirúrgica/efectos adversosRESUMEN
OBJECTIVE: Chronic inflammation has earlier been detected in ruptured intracranial aneurysms. A previous study detected both dental bacterial DNA and bacterial-driven inflammation in ruptured intracranial aneurysm walls. The aim of this study was to compare the presence of oral and pharyngeal bacterial DNA in ruptured and unruptured intracranial aneurysms. The hypothesis was that oral bacterial DNA findings would be more common and the amount of bacterial DNA would be higher in ruptured aneurysm walls than in unruptured aneurysm walls. MATERIALS AND METHODS: A total of 70 ruptured (n = 42) and unruptured (n = 28) intracranial aneurysm specimens were obtained perioperatively in aneurysm clipping operations. Aneurysmal sac tissue was analysed using a real-time quantitative polymerase chain reaction to detect bacterial DNA from several oral species. Both histologically non-atherosclerotic healthy vessel wall obtained from cardiac by-pass operations (LITA) and arterial blood samples obtained from each aneurysm patient were used as control samples. RESULTS: Bacterial DNA was detected in 49/70 (70%) of the specimens. A total of 29/42 (69%) of the ruptured and 20/28 (71%) of the unruptured aneurysm samples contained bacterial DNA of oral origin. Both ruptured and unruptured aneurysm tissue samples contained significantly more bacterial DNA than the LITA control samples (p-values 0.003 and 0.001, respectively). There was no significant difference in the amount of bacterial DNA between the ruptured and unruptured samples. CONCLUSION: Dental bacterial DNA can be found using a quantitative polymerase chain reaction in both ruptured and unruptured aneurysm walls, suggesting that bacterial DNA plays a role in the pathogenesis of cerebral aneurysms in general, rather than only in ruptured aneurysms.
Asunto(s)
Aneurisma Roto/microbiología , ADN Bacteriano/aislamiento & purificación , Aneurisma Intracraneal/microbiología , Boca/microbiología , Aggregatibacter actinomycetemcomitans/genética , Femenino , Fusobacterium nucleatum/genética , Bacilos Gramnegativos Anaerobios Rectos, Curvos y Espirales/genética , Humanos , Masculino , Persona de Mediana Edad , Peptostreptococcus/genética , Faringe/microbiología , Porphyromonas gingivalis/genética , Prevotella intermedia/genética , Staphylococcus aureus/genética , Staphylococcus epidermidis/genética , Streptococcus anginosus/genética , Streptococcus gordonii/genética , Streptococcus mitis/genética , Streptococcus oralis/genética , Streptococcus sanguis/genética , Diente/microbiología , Treponema denticola/genéticaRESUMEN
Streptococcus anginosus is an emerging pathogen, but little is known about its virulence factors. To detect the genes responsible for ß-haemolysis we performed genomic mutagenesis of the ß-haemolytic S. anginosus type strain ATCC 12395 using the vector pGhost9:ISS1. Integration site analysis of 15 non-haemolytic mutants identified a gene cluster with high homology to the genes of the streptolysin S (SLS) encoding sag gene cluster of S. pyogenes. The gene cluster harbours 10 open reading frames displaying significant similarities to the S. pyogenes genes sagA-sagI, with the identities on protein level ranging from 38 to 87%. Complementation assays of S. anginosus sagB and sagD integration mutants with the respective genes confirmed their importance for ß-haemolysin production and suggest the presence of post-translational modifications in S. anginosus SLS similar to SLS of S. pyogenes. Characterization of the S. anginosus haemolysin in comparison to the S. pyogenes SLS showed that the haemolysin is surface bound, but in contrast to S. pyogenes neither fetal calf serum nor RNA was able to stabilize the haemolysin of S. anginosus in culture supernatants. Inhibition of ß-haemolysis by polyethylene glycol of different sizes was carried out, giving no evidence of a pore-forming haemolytic mechanism. Analysis of a whole genome shotgun sequence of Streptococcus constellatus, a closely related streptococcal species that belongs to the S. anginosus group, revealed a similar sag gene cluster. Employing a genomic mutagenesis strategy we were able to determine an SLS encoding gene cluster in S. anginosus and demonstrate its importance for ß-haemolysin production in S. anginosus.
Asunto(s)
Proteínas Bacterianas/genética , Genes Bacterianos/genética , Proteínas Hemolisinas/genética , Streptococcus anginosus/genética , Proteínas Bacterianas/antagonistas & inhibidores , Escherichia coli/genética , Vectores Genéticos/genética , Proteínas Hemolisinas/antagonistas & inhibidores , Hemólisis/genética , Humanos , Familia de Multigenes/genética , Mutagénesis/genética , Mutación/genética , Sistemas de Lectura Abierta/genética , Plásmidos/genética , Polietilenglicoles/farmacología , Regiones Promotoras Genéticas/genética , Procesamiento Proteico-Postraduccional/genética , Homología de Secuencia , Streptococcus constellatus/genética , Streptococcus pyogenes/genética , Estreptolisinas/genética , Factores de Virulencia/genéticaRESUMEN
OBJECTIVE: To elucidate a possible involvement of Streptococcus anginosus in oral cancer, we assessed the frequency of S. anginosus infection in oral cancer tissues, and investigated its infection route. MATERIALS AND METHOD: The tissue specimens were obtained from 46 oral cancer and three precancerous leukoplakia subjects. Frequency of S. anginosus infection was assessed by a species-specific polymerase chain reaction (PCR) assay. The genotype of the clinical isolates taken from cancer tissue and dental plaque samples was analyzed using pulsed-field gel electrophoresis (PFGE). RESULTS: S. anginosus DNA was frequently detected in squamous cell carcinoma (19/42), but not in other types of cancer (lymphoma and rhabdomyosarcoma) or leukoplakia samples. A subject-based analysis revealed that S. anginosus was solely detected in dental plaque and not in saliva from all 19 S. anginosus-positive squamous cell carcinoma cases. Further, the genotype of S. anginosus isolated from cancer tissue was identical to that from dental plaque of the same patients. CONCLUSION: Infection of S. anginosus could occur frequently in oral squamous cell carcinoma and that dental plaque could be a dominant reservoir of the S. anginosus.
Asunto(s)
Carcinoma de Células Escamosas/microbiología , Neoplasias de la Boca/microbiología , Infecciones Estreptocócicas/complicaciones , Streptococcus anginosus/aislamiento & purificación , Anciano , Animales , Placa Dental/microbiología , Electroforesis en Gel de Campo Pulsado , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa/normas , Saliva/microbiología , Sensibilidad y Especificidad , Streptococcus anginosus/genéticaRESUMEN
It is difficult to distinguish mutans streptococci on the species level, and even more so on the subspecies level. Intact cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) (ICM) was applied to reference strains of five of the species of the mutans group (Streptococcus criceti, Streptococcus downei, Streptococcus mutans, Streptococcus ratti, Streptococcus sobrinus), nonmutans streptococci (Streptococcus oralis, Streptococcus mitis, Streptococcus salivarius, and Streptococcus sanguinis), and 177 mutans streptococci isolated from saliva of 10 children. From the analysis of the reference strains, readily distinguishable ICM mass spectra were obtained for the different species. Based on multivariate statistical analysis, a correct and unambiguous assignment was made of the spectra of 159 isolated mutans streptococci to S. mutans and 16 isolates to S. sobrinus. Two isolates were sorted out and were identified by sequencing of their 16S rRNA genes as Streptococcus anginosus. In addition, ICM indicated a misclassification for some reference strains (AHT, V 100 and E 49) and re-classified AHT and E 49 as S. ratti and V 100 as S. sobrinus. This was confirmed by 16S rDNA sequencing. Based on a statistical similarity analysis of the spectra of reference strains and a quantitative assessment of the reproducibility of ICM, the isolates identified as either S. mutans or S. sobrinus were phenotyped on the subspecies level. In the population of the clinical isolates, 14 unambiguously different S. mutans and three different S. sobrinus phenotypes were detected. ICM proved to be a powerful tool for a differentiation of mutans streptococci down to the subspecies level.
Asunto(s)
Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Streptococcus/clasificación , Niño , Humanos , Fenotipo , ARN Ribosómico 16S/genética , Reproducibilidad de los Resultados , Saliva/microbiología , Análisis de Secuencia de ADN , Análisis de Secuencia de ARN , Streptococcus/genética , Streptococcus anginosus/clasificación , Streptococcus anginosus/genética , Streptococcus mitis/clasificación , Streptococcus mitis/genética , Streptococcus mutans/clasificación , Streptococcus mutans/genética , Streptococcus oralis/clasificación , Streptococcus oralis/genética , Streptococcus sobrinus/clasificación , Streptococcus sobrinus/genéticaRESUMEN
Multiple cancers frequently occur in the upper aerodigestive tract. The high incidence rate of multiple carcinomas in this region is often explained in terms of involvement of the same underlying risk factors. It has been reported that the oral bacterium Streptococcus anginosus (S. anginosus) is associated with esophageal, gastric, and pharyngeal cancer tissues. In this study, a highly specific quantification method for S. anginosus DNA using real-time PCR was established. We employed this assay to determine whether S. anginosus is also associated with oral cancer tissues. This precise quantification method revealed different degrees of infection with S. anginosus in esophageal cancer and oral cancer. We assayed 10 ng of genomic DNA from cancer tissues, and found that eight of 18 samples (44%) from the esophagus contained a detectable level (>10 fg) of S. anginosus DNA, whereas this was the case for only five of 38 samples (13%) of oral cancer. The quantity of S. anginosus DNA in the esophageal cancer tissues was significantly higher than in oral cancer. The maximum amount of S. anginosus DNA was approximately ten times higher in esophageal than in oral cancer tissues. In addition, none of the five different oral cancer sites (floor of the mouth, mandibular gingival, maxillary gingival, buccal mucosal, and tongue) showed significant signs of S. anginosus infection. On the other hand, most non-cancerous tissues of the esophagus and tongue showed an undetectable level of S. anginosus. These results suggest that S. anginosus is associated with esophageal cancer, but is not closely related with oral cancer.
Asunto(s)
ADN Bacteriano/análisis , Neoplasias Esofágicas/microbiología , Neoplasias de la Boca/microbiología , Infecciones Estreptocócicas/microbiología , Streptococcus anginosus/genética , Secuencia de Bases , Cartilla de ADN/química , ADN de Neoplasias/genética , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Streptococcus anginosus/aislamiento & purificaciónRESUMEN
The purpose of this study was to survey the prevalence of streptococcal species, especially Streptococcus anginosus (which has been reported to be associated with cancer in the upper digestive tract), Streptococcus constellatus, and Streptococcus intermedius in the saliva of different age groups. A sequence analysis of 16S rDNA was performed and DNA quantified using real-time polymerase chain reaction. The S. anginosus level increased with age, whereas the levels of S. constellatus and S. intermedius did not change. Streptococcus mitis was the predominant species in the saliva of all the age groups but, unlike the S. anginosus, the proportion of S. mitis in the salivary bacteria decreased with age. The increase in S. anginosus with age should be carefully monitored because of its association with diseases, including cancer.