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1.
Int J Syst Evol Microbiol ; 70(2): 732-737, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31702538

RESUMO

An obligately anaerobic, Gram-stain-positive and coccus-shaped bacterium, designated strain YH-panp20T, was isolated from pig faeces. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that the isolate belongs to the family Erysipelotrichaceae, and is most closely related to Catenisphaera adipataccumulans KCTC 15517T (93.5 % sequence similarity), followed by Faecalitalea cylindroides KCTC 5815T (92.2 %), Faecalicoccus acidiformans KCTC 15521T (90.2 %) and Holdemanella biformis KCTC 5969T (89.6 %). Average nucleotide identity values between YH-panp20T and its closest relatives were lower than 71 %. The G+C content of the isolate was 38.4 mol%, and its cell-wall peptidoglycan was found to be of A1γ type and contained meso-diaminopimelic acid. The predominant fatty acids were C18 : 1 cis 9, C18 : 0 DMA and C16 : 0. The major end-products of glucose fermentation were lactate, acetate and formate. Therefore, based on the phenotypic, phylogenetic and chemotaxonomic properties, a novel genus and species, Absicoccus porci gen. nov., sp. nov., is proposed for isolate YH-panp20T (=KCTC 15747T=JCM 32769T).


Assuntos
Firmicutes/classificação , Filogenia , Suínos/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , Parede Celular/química , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Fezes/microbiologia , Fermentação , Firmicutes/isolamento & purificação , Bacilos Gram-Positivos/genética , Peptidoglicano/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA
2.
Artigo em Japonês | MEDLINE | ID: mdl-31856572

RESUMO

Identification of bacteria by using MALDI Biotyper is relevant at the species category if Score Value (SV) is not less than 2.000. However, in practical examination, the analysis by MALDI Biotyper frequently produces the multiple candidate bacterial species with SV ≥2.000. In this study, we analyzed the ratio of multiple results among 10,081 specimens and identified the species of bacteria with high frequency of multiple results. Our analysis indicated that 8,129 strains out of 10,081 strains examined from July 2015 to July 2017, showed multiple identification results with MALDI Biotyper, and that multiple result was obtained in 4.9% of gram positive cocci analysis, 5.8% of gram positive rods, 25.4% of gram negative cocci, 16% of gram negative rod, none of fungus. In particular, MALDI Biotyper analysis of Enterobacter spp. (E. cloacae, E. asburiae, E. kobei, etc.), Acinetobacter spp. (A. baumannii, A. nosocomialis, A. pittii etc.), Neisseria spp. (N. flavescens, N. perflava etc.) had high ratios of multiple results. Our data suggests that genetic homology among bacteria results in multiple results of bacteria identification. The mass spectrometer method is the rapid test for bacteria identification. However, for obtaining higher specificity, it is required to combine with other methods. Furthermore, systematic annotation of bacteria is highly recommended.


Assuntos
Bactérias Gram-Negativas , Cocos Gram-Positivos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Bacilos Gram-Positivos , Reprodutibilidade dos Testes
3.
Oper Neurosurg (Hagerstown) ; 14(6): 681-685, 2018 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-28961750

RESUMO

BACKGROUND: Intracavernous aneurysms constitute up to 9% of all intracranial aneurysms and 6% are infectious (IIA). First line therapy is a protracted antibiotic course, yet with failure, surgery and endovascular parent vessel sacrifice have been utilized. Reconstructive endovascular therapies have emerged for aneurysm control and may demonstrate a safer therapeutic alternative. OBJECTIVE: To present an IIA treated with a flow-diverting Pipeline stent (ev3 Neurovascular, Irvine, California). METHODS: A 41-yr-old female presented with visual loss, ophthalmoplegia, and cavernous sinus thrombosis with an associated phlegmon. Transsphenoidal evacuation was performed without complication or bleeding and she continued on medical therapy. Two weeks postoperatively, she developed a worsening right third cranial nerve palsy and MRA demonstrated a 1-cm right IIA, not evident on postoperative MRI. Three days of dual antiplatelet therapy preceded successful pipeline embolization. Angiography demonstrated aneurysm obliteration at 3 mo and her right ophthalmoplegia resolved. RESULTS: A literature review identified 6 reported cases of IIAs treated with stent embolization. Only 1 documented a flow-diverting Silk stent used in a child. All lesions were obliterated at follow-up without neurological sequelae. No complication arose with implantation in the setting of infection, and as few as 3 d of dual antiplatelet therapy was sufficient for preprocedural prophylaxis, although in Vivo antiplatelet activity may be more significant. CONCLUSION: We report the first case of an IIA treated with a flow-diverting pipeline stent. These devices preserve native vasculature and neurological function compared to surgical and endovascular vessel sacrifice strategies. They appear to be safe management options for the treatment of IIAs.


Assuntos
Actinomicose/complicações , Aspergilose/complicações , Doenças das Artérias Carótidas/terapia , Embolização Terapêutica/métodos , Procedimentos Endovasculares/métodos , Infecções por Bactérias Gram-Positivas/complicações , Aneurisma Intracraniano/terapia , Stents , Adulto , Doenças das Artérias Carótidas/diagnóstico por imagem , Doenças das Artérias Carótidas/microbiologia , Trombose do Corpo Cavernoso/etiologia , Celulite (Flegmão)/etiologia , Celulite (Flegmão)/microbiologia , Celulite (Flegmão)/cirurgia , Descompressão Cirúrgica , Embolização Terapêutica/instrumentação , Emergências , Procedimentos Endovasculares/instrumentação , Desenho de Equipamento , Feminino , Bacilos Gram-Positivos/isolamento & purificação , Humanos , Aneurisma Intracraniano/diagnóstico por imagem , Aneurisma Intracraniano/microbiologia , Angiografia por Ressonância Magnética , Oftalmoplegia/etiologia , Tomografia Computadorizada por Raios X
4.
Foodborne Pathog Dis ; 14(3): 177-187, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-28128639

RESUMO

A study was conducted to evaluate Sensititre® Automated Reading and Incubation System 2x System (ARIS), API® (API), and Bruker MALDI-TOF MS (MALDI) bacterial species identification systems using 132 diverse bacterial isolates from bovine milk samples and bulk tank milk received at the Penn State Animal Diagnostic Laboratory. The results were compared with 16S rRNA gene sequence analysis, which served as the reference method for species identification. The ARIS, API, and MALDI identified 0%, 40%, and 33.4% of species classified as Gram-positive rod isolates belonging to genera Arthrobacter, Bacillus, Brachybacterium, Brevibacterium, and Corynebacterium, respectively. It was observed that 76.5%, 93.9%, and 96.9% of catalase-negative, Gram-positive cocci (n = 33; Aerococcus, Enterococcus, Lactococcus, Streptococcus) were correctly identified to the species level by ARIS, API, and MALDI, respectively, while 33.4%, 84.5%, and 97.7% of catalase-positive, Gram-positive cocci (n = 45; Kocuria, Staphylococcus) were correctly identified to their species by ARIS, API, and MALDI, respectively. A total of 48 isolates (Acinetobacter, Citrobacter, Enterobacter, Escherichia, Klebsiella, Pantoea, Pasteurella, Providencia, Pseduomonas, Serratia) of Gram-negative bacteria were examined, of which 85.4%, 93.7%, and 95.8% of the isolates were correctly identified to the species level by ARIS, API, and MALDI, respectively. In our laboratory, the MALDI had the least costs associated with consumables and reagents compared to ARIS, API, and 16S rRNA identification methods. Identification of bacterial species was accomplished in <2 h using MALDI and 24 h for ARIS, API, and 16S rRNA identification systems.


Assuntos
Bactérias Gram-Negativas/isolamento & purificação , Cocos Gram-Positivos/isolamento & purificação , Bacilos Gram-Positivos/isolamento & purificação , Mastite Bovina/diagnóstico , Leite/microbiologia , Animais , Bovinos , Feminino , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Bactérias Gram-Negativas/classificação , Cocos Gram-Positivos/classificação , Bacilos Gram-Positivos/classificação , Mastite Bovina/microbiologia , RNA Ribossômico 16S/isolamento & purificação , Análise de Sequência de RNA , Especificidade da Espécie , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
5.
J Clin Lab Anal ; 31(5)2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27859671

RESUMO

OBJECTIVES: To evaluate the ability of laser flow cytometry to predict cocci/mixed growth in the pre-analytical phase of urine specimens. METHODS: We retrospectively reviewed urine samples from women with uncomplicated urinary tract infections from urologic clinics for study. Urine analyses were performed with laser flow cytometry (UF1000i, Sysmex, Kobe, Japan) and then diagrams were generated (forward scatter vs. fluorescent light scatter). Each specimen (bacteria count >357 BACT/µL) was classified as either cocci bacteria or rods/mixed growth according to the diagrams. Standard urine cultures were performed, and the agreement between cultures and the UF1000i interpretations was analyzed with kappa statistics. RESULTS: Finally, 491 specimens met the criteria for analysis. Among the 376 specimens with single bacteria growth, there were 26 gram-positive cocci (13 Streptococci spp., 7 Staphylococci spp., 6 Enterococci spp.), 1 gram-positive rods (Corynebacterium spp.), and 349 gram-negative rods (273 Escherichia coli, 33 Klebsiella spp., 29 Proteus spp., 6 Citrobacter spp., 4 Enterobacter spp., 3 Pseudomonas spp., and 1 Providencia spp.). There were 115 specimens with two bacteria species or more that were regarded as mixed growth. Agreement of rods or cocci/mixed growth between the laser flow cytometry and urine cultures yielded a kappa value of 0.58. The positive and negative predictive rate of the UF1000i for cocci/mixed growth in voided urine culture was 81.8% and 84.7%, respectively. CONCLUSIONS: Through laser flow cytometry, we can predict growth of cocci/mixed growth in the pre-analytical phase of urine culture, thus avoiding unnecessary urine culture and waiting time.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Coinfecção/microbiologia , Citometria de Fluxo/métodos , Cocos Gram-Positivos/citologia , Bacilos Gram-Positivos/citologia , Infecções Urinárias/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Coinfecção/diagnóstico , Feminino , Cocos Gram-Positivos/isolamento & purificação , Bacilos Gram-Positivos/isolamento & purificação , Humanos , Pessoa de Meia-Idade , Estudos Retrospectivos , Infecções Urinárias/diagnóstico
6.
Insect Biochem Mol Biol ; 81: 62-71, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-28017798

RESUMO

Cqm1 and Aam1 are α-glucosidases (EC 3.2.1.20) expressed in Culex quinquefasciatus and Aedes aegypti larvae midgut, respectively. These orthologs share high sequence similarity but while Cqm1 acts as a receptor for the Binary (Bin) insecticidal toxin from Lysinibacillus sphaericus, Aam1 does not bind the toxin, rendering Ae. aegypti refractory to this bacterium. Aam1 is heavily glycosylated, contrasting to Cqm1, but little is known regarding how glycosylation impacts on its function. This study aimed to compare the N-glycosylation patterns and the catalytic activities of Aam1 and Cqm1. Mutant proteins were generated where predicted Aam1 N-glycosylation sites (N-PGS) were either inserted into Cqm1 or abrogated in Aam1. The mutants validated four N-PGS which were found to localize externally on the Aam1 structure. These Aam1 and Cqm1 mutants maintained their Bin binding properties, confirming that glycosylation has no role in this interaction. The α-glucosidase activity of both proteins was next investigated, with Aam1 having a remarkably higher catalytic efficiency, influenced by changes in glycosylation. Molecular dynamics showed that glycosylated and nonglycosylated Aam1 models displayed distinct patterns that could influence their catalytic activity. Differential N-glycosylation may then be associated with higher catalytic efficiency in Aam1, enhancing the functional diversity of related orthologs.


Assuntos
Aedes/enzimologia , Culex/enzimologia , alfa-Glucosidases/metabolismo , Animais , Glicosilação , Bacilos Gram-Positivos , Simulação de Dinâmica Molecular
7.
J Clin Microbiol ; 54(12): 2928-2936, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27629905

RESUMO

Commensal bacteria from the skin and mucosal surfaces are routinely isolated from patient samples and considered contaminants. The majority of these isolates are catalase-positive Gram-positive rods from multiple genera routinely classified as diphtheroids. These organisms can be seen upon Gram staining of clinical specimens or can be isolated as the predominant or pure species in culture, raising a priori suspicion of a possible involvement in infection. With the development and adoption of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), suspicious isolates are now routinely identified to the species level. In this study, we performed a retrospective data review (2012 to 2015) and utilized site-specific laboratory criteria and chart reviews to identify species within the diphtheroid classification representative of true infection versus contamination. Our data set included 762 isolates from 13 genera constituting 41 bacterial species. Only 18% represented true infection, and 82% were deemed contaminants. Clinically significant isolates were identified in anaerobic wounds (18%), aerobic wounds (30%), blood (5.5%), urine (22%), cerebrospinal fluid (24%), ophthalmologic cultures (8%), and sterile sites (20%). Organisms deemed clinically significant included multiple Actinomyces species in wounds, Propionibacterium species in joints and cerebrospinal fluid associated with central nervous system hardware, Corynebacterium kroppenstedtii (100%) in breast, and Corynebacterium striatum in multiple sites. Novel findings include clinically significant urinary tract infections by Actinomyces neuii (21%) and Corynebacterium aurimucosum (21%). Taken together, these findings indicate that species-level identification of diphtheroids isolated with a priori suspicion of infection is essential to accurately determine whether an isolate belongs to a species associated with specific types of infection.


Assuntos
Bacilos Gram-Positivos/classificação , Bacilos Gram-Positivos/isolamento & purificação , Tipagem Molecular/métodos , Membrana Mucosa/microbiologia , Pele/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Bacilos Gram-Positivos/genética , Humanos , RNA Ribossômico 16S/genética , Estudos Retrospectivos , Análise de Sequência de DNA
8.
Diagn Microbiol Infect Dis ; 85(3): 372-376, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27179368

RESUMO

With the advent of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), most Gram-positive rods (GPRs) are readily identified; however, their clinical relevance in blood cultures remains unclear. Herein, we assessed the clinical significance of GPRs isolated from blood and identified in the era of MALDI-TOF MS. A retrospective chart review of patients presenting to the Mayo Clinic, Rochester, MN, from January 1, 2013, to October 13, 2015, was performed. Any episode of a positive blood culture for a GPR was included. We assessed the number of bottles positive for a given isolate, time to positivity of blood cultures, patient age, medical history, interpretation of culture results by the healthcare team and whether infectious diseases consultation was obtained. We also evaluated the susceptibility profiles of a larger collection of GPRs tested in the clinical microbiology laboratory of the Mayo Clinic, Rochester, MN from January 1, 2013, to October 31, 2015. There were a total of 246 GPRs isolated from the blood of 181 patients during the study period. 56% (n = 101) were deemed contaminants by the healthcare team and were not treated; 33% (n = 59) were clinically determined to represent true bacteremia and were treated; and 8% (n = 14) were considered of uncertain significance, with patients prescribed treatment regardless. Patient characteristics associated with an isolate being treated on univariate analysis included younger age (P = 0.02), identification to the species level (P = 0.02), higher number of positive blood culture sets (P < 0.0001), lower time to positivity (P < 0.0001), immunosuppression (P = 0.03), and recommendation made by an infectious disease consultant (P = 0.0005). On multivariable analysis, infectious diseases consultation (P = 0.03), higher number of positive blood culture sets (P = 0.0005) and lower time to positivity (P = 0.03) were associated with an isolate being treated. 100, 83, 48 and 34% of GPRs were susceptible to vancomycin, meropenem, penicillin and ceftriaxone, respectively.


Assuntos
Antibacterianos/farmacologia , Bacteriemia/microbiologia , Sangue/microbiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Bacilos Gram-Positivos/efeitos dos fármacos , Bacilos Gram-Positivos/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Antibacterianos/uso terapêutico , Bacteriemia/tratamento farmacológico , Bacteriemia/epidemiologia , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Infecções por Bactérias Gram-Positivas/epidemiologia , Bacilos Gram-Positivos/química , Humanos , Testes de Sensibilidade Microbiana , Estudos Retrospectivos
9.
Int J Syst Evol Microbiol ; 66(6): 2225-2233, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26961808

RESUMO

A novel Gram-stain-negative, rod-shaped, facultatively anaerobic, oxidase-negative and catalase-positive bacterium, designated 2W32T, was isolated from a marine solar saltern on the coast of Weihai, Shandong Province, China. Strain 2W32T was tolerant to moderate salt conditions. Optimal growth occurred at 33-37 °C (range 20-45 °C) and pH 7.5-8.0 (range pH 7.0-8.5) with 6-10 % (w/v) NaCl (range 2-18 %). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 2W32T shared highest similarity with Aliifodinibius sediminis YIM J21T (94.6 %), Aliifodinibius roseus YIM D15T (94.4 %), Fodinibius salinus YIM C003T (93.6 %), Gracilimonas tropica CL-CB462T (88.6 %) and Balneola vulgaris 13IX/A01/164T (86.4 %) and less than 83.0 % similarity with other species of the phylum Bacteroidetes. The isolate and closely related species formed a novel family-level clade in the phylum Bacteroidetes. The polar lipid profile of the novel isolate consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unidentified aminolipid, an unidentified glycolipid and an unidentified lipid. The dominant cellular fatty acids (>10 %) were iso-C15 : 0, iso-C17 : 1ω9c and summed feature 3 (C16:1ω7c and/or iso-C15 : 0 2-OH) and the sole respiratory quinone was menaquinone 7 (MK-7). The DNA G+C content of strain 2W32T was 47.5 mol %. Comparative analysis of 16S rRNA gene sequences and characterization indicated that strain 2W32T represents a novel species within the genus Aliifodinibius, for which the name Aliifodinibius halophilus sp. nov. is proposed. The type strain is 2W32T (=KCTC 42497T=CICC 23869T). In addition, a novel family, Balneolaceae fam. nov., is proposed to accommodate the genera Fodinibius, Aliifodinibius, Gracilimonas and Balneola.


Assuntos
Bacteroidetes/classificação , Bacilos Gram-Positivos/classificação , Filogenia , Salinidade , Microbiologia da Água , Técnicas de Tipagem Bacteriana , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Bacilos Gram-Positivos/genética , Bacilos Gram-Positivos/isolamento & purificação , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
10.
Braz J Microbiol ; 46(2): 443-53, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26273259

RESUMO

The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240) thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5%) represented by B. licheniformis (119), B. aerius (44), B. sonorensis (33), B. subtilis (subsp. spizizenii (2) and subsp. inaquosurum (6)), B. amyloliquefaciens (subsp. amyloliquefaciens (4) and subsp. plantarum (4)), B. tequilensis (3), B. pumilus (3) and Bacillus sp. (19). Only six isolates (2.5%) belonged to the genus Aeribacillus represented by A. pallidus (4) and Aeribacillus sp. (2). In this study, B. aerius and B. tequilensis are described for the first time as thermophilic bacteria. Moreover, 71.25%, 50.41% and 5.41% of total strains exhibited high amylolytic, proteolytic or cellulolytic activity respectively.


Assuntos
Bacillaceae/classificação , Bacillaceae/isolamento & purificação , Bacilos Gram-Positivos/classificação , Bacilos Gram-Positivos/isolamento & purificação , Fontes Termais/microbiologia , Microbiologia do Solo , Microbiologia da Água , Bacillaceae/genética , Bacillaceae/efeitos da radiação , Biodiversidade , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Bacilos Gram-Positivos/genética , Bacilos Gram-Positivos/efeitos da radiação , Dados de Sequência Molecular , Marrocos , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Esporos Bacterianos/citologia
11.
Int J Syst Evol Microbiol ; 65(Pt 1): 293-7, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25342112

RESUMO

A novel thermophilic bacterial strain, CBS-Z(T), was isolated from a terrestrial hot spring in the Changbai Mountains, PR China. Cells of strain CBS-Z(T) were short straight rods without flagella and had Gram-positive cell walls. Growth was observed at 40-90 °C (optimum 75 °C) and at pH 5.6-8.6 (optimum pH 7.8). The primary end-products from the fermentation of filter paper by strain CBS-Z(T) were acetate, lactate, H2, and CO2. The main cellular fatty acids were iso-C17:0, iso-C14:0 3-OH and C16:0. The G+C content of the genomic DNA was 36.08 mol%. Multiple sequence alignment of the 16S rRNA gene sequence and phylogenetic analyses indicated that strain CBS-Z(T) belongs to the genus Caldicellulosiruptor and the most similar micro-organism was Caldicellulosiruptor saccharolyticus DSM 8903(T) (96.36% 16S rRNA gene sequence similarity); the 16S rRNA gene sequence similarity of strain CBS-Z(T) to other species was below 95%. Based on its phylogenetic and phenotypic characteristics, strain CBS-Z(T) represents a novel species of the genus Caldicellulosiruptor, for which the name Caldicellulosiruptor changbaiensis sp. nov. is proposed. The type strain is CBS-Z(T) ( =DSM 26941(T) =CGMCC 1.5180(T)).


Assuntos
Bacilos Gram-Positivos/classificação , Fontes Termais/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Bacilos Gram-Positivos/genética , Bacilos Gram-Positivos/isolamento & purificação , Hidrogênio/metabolismo , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Int J Syst Evol Microbiol ; 65(Pt 3): 805-10, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25481294

RESUMO

An obligately anaerobic bacterium, designated strain GK12(T), was isolated from an anaerobic digester in Fukagawa, Hokkaido Prefecture, Japan. The cells of strain GK12(T) were non-motile, non-spore-forming cocci that commonly occurred in chains. 16S rRNA gene sequence analysis revealed that strain GK12(T) was affiliated with the family Erysipelotrichaceae in the phylum Firmicutes and showed 91.8 % sequence similarity to the most closely related species, Faecalicoccus acidiformans. The strain grew at 30-50 °C (optimally at 40 °C) and at pH 5.5-8.5 (optimally at pH 7.5). The main end product of glucose fermentation was lactate. Yeast extract was required for growth. The strain contained C14 : 0, C14 : 0 1,1-dimethoxyalkane (DMA), C16 : 0 DMA and C18 : 0 DMA as the major cellular fatty acids (>10 % of the total). The polar lipid profile was composed of phosphatidylglycerol, phosphatidylinositol and an unidentified phospholipid. The whole-cell sugars were galactose, rhamnose and ribose. The cell-wall murein contained alanine, glutamic acid, lysine, serine and threonine, but not diaminopimelic acid. The G+C content of the genomic DNA was 47.7 mol%. Based on phenotypic, phylogenetic and chemotaxonomic properties, a novel genus and species, Catenisphaera adipataccumulans gen. nov., sp. nov., is proposed to accommodate strain GK12(T) ( = NBRC 108915(T) = DSM 25799(T)).


Assuntos
Bactérias Anaeróbias/classificação , Bacilos Gram-Positivos/classificação , Filogenia , Bactérias Anaeróbias/genética , Bactérias Anaeróbias/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , Reatores Biológicos/microbiologia , DNA Bacteriano/genética , Ácidos Graxos/química , Fermentação , Bacilos Gram-Positivos/genética , Bacilos Gram-Positivos/isolamento & purificação , Japão , Dados de Sequência Molecular , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
13.
J Periodontal Res ; 50(3): 320-9, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25040261

RESUMO

BACKGROUND AND OBJECTIVES: A number of species/phylotypes have been newly implicated as putative periopathogens. The objective of this study was to explore associations among classical and new pathogens in subgingival biofilm and to assess their relative importance to chronic periodontitis. MATERIAL AND METHODS: Pooled subgingival biofilm samples were obtained from 40 patients with chronic periodontitis and 40 healthy controls. Taqman q-PCR assays were used to determine the absolute and relative counts of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Parvimonas micra, Filifactor alocis, oral Synergistetes and oral TM7s. Microbial associations were assessed using cluster analysis. Different statistical models were used to explore associations between microbial parameters and periodontitis. RESULTS: The median log and relative counts were lowest for TM7s (4.4 and 0.0016%, respectively) and highest for oral Synergistetes (7.2 and 1.4%, respectively). Oral Synergistetes clustered strongly with the red complex, particularly T. forsythia (100% rescaled similarity). All species/phylotypes except TM7s were significantly associated with periodontitis (Mann-Whitney test; p ≤ 0.005). However, P. gingivalis and F. alocis lost association after adjusting for confounders (ordinal regression). In receiving operator characteristic curve analysis, the log counts of oral Synergistetes were the best markers of periodontitis (82.5% sensitivity and specificity), followed by those of T. forsythia, P. micra and T. denticola. In prediction analysis, however, P. micra was the only microbial predictor of periodontal parameters. CONCLUSIONS: Oral Synergistetes are presented here as new members of the red complex, with relative importance to periodontitis exceeding that of the classical members. P. micra is shown as an important periodontal pathogen warranting more attention.


Assuntos
Biofilmes , Periodontite Crônica/microbiologia , Placa Dentária/microbiologia , Gengiva/microbiologia , Adulto , Área Sob a Curva , Carga Bacteriana , Bacteroides/isolamento & purificação , Estudos de Casos e Controles , Índice de Placa Dentária , Feminino , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/isolamento & purificação , Bacilos Gram-Positivos/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Peptostreptococcus/isolamento & purificação , Perda da Inserção Periodontal/microbiologia , Índice Periodontal , Porphyromonas gingivalis/isolamento & purificação , Curva ROC , Sensibilidade e Especificidade , Treponema denticola/isolamento & purificação
14.
Int J Syst Evol Microbiol ; 65(Pt 2): 570-7, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25404482

RESUMO

A thermophilic, agar-degrading bacterium, strain FAB2(T), was isolated from sewage sludge compost. According to phylogenetic analysis based on 16S rRNA gene sequences, strain FAB2(T) belonged to the family Paenibacillaceae within the phylum Firmicutes. However, FAB2(T) was different enough at the genus level from closely related species. The percentages of 16S rRNA gene sequence similarity with related organisms were 90.4 % for Thermobacillus xylanilyticus, 91.8 % for Paenibacillus barengoltzii, 89.4 % for Cohnella lupini, 90.1 % for Fontibacillus aquaticus, and 89.0 % for Saccharibacillus sacchari. Morphological and physiological analyses revealed that the strain was motile, rod-shaped, Gram-stain-positive, aerobic and able to form oval endospores in swollen sporangia. Ammonium was required as a nitrogen source while nitrate, nitrite, urea and glutamate were not utilized. Catalase and oxidase activities were weakly positive and positive, respectively. The bacterium grew in the temperature range of 50-65 °C and in media with pH 7.5 to 9.0. Optimal growth occurred at 60 °C and pH 8.0-8.6. Growth was inhibited at pH≤7.0 and NaCl concentrations ≥2.5 % (w/v). In chemotaxonomic characterization, MK-7 was identified as the dominant menaquinone. Major fatty acids were iso-C16 : 0 and C16 : 0. Dominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Phosphatidylcholine was present in a moderate amount. The diamino acid in the cell wall was meso-diaminopimelic acid. The G+C content of the genomic DNA was 49.5 mol% in a nucleic acid study. On the basis of genetic and phenotypic characteristics, strain FAB2(T) ( = NBRC 109510(T) = KCTC 33130(T)) showed characteristics suitable for classification as the type strain of a novel species of a new genus in the family Paenibacillaceae, for which the name Ammoniibacillus agariperforans gen. nov., sp. nov. is proposed.


Assuntos
Ágar/metabolismo , Bacillales/classificação , Filogenia , Microbiologia do Solo , Bacillales/genética , Bacillales/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Bacilos Gram-Positivos/genética , Bacilos Gram-Positivos/crescimento & desenvolvimento , Bacilos Gram-Positivos/isolamento & purificação , Japão , Dados de Sequência Molecular , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
15.
J Dent Res ; 93(8): 725-32, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24898946

RESUMO

Infection-induced periodontal disease has been primarily focused on a small group of periodontal pathogens. A paradigm shift, based on data emerging from the oral microbiome project, now suggests the involvement of as-yet-unculturable and fastidious organisms. Collectively, these studies have demonstrated that there are changes in the periodontal status associated with shifts in the composition of the bacterial community in the periodontal pocket. In addition, it is likely that the emerging new pathogens may play a more significant role in the disease. One of the organisms previously unrecognized is Filifactor alocis. While this Gram-positive anaerobic rod has been identified in peri-implantitis, in endodontic infections, and in patients with localized aggressive periodontitis, its presence is now observed at significantly higher levels in patients with adult periodontitis or refractory periodontitis. Its colonization properties and its potential virulence attributes support the proposal that F. alocis should be included as a diagnostic indicator of periodontal disease. Moreover, these emerging characteristics would be consistent with the polymicrobial synergy and dysbiosis (PSD) periodontal pathogenesis model. Here, unique characteristics of F. alocis are discussed. F. alocis has specific factors that can modulate multiple changes in the microbial community and host cell proteome. It is likely that such variations at the molecular level are responsible for the functional changes required to mediate the pathogenic process.


Assuntos
Infecções por Bactérias Gram-Positivas/diagnóstico , Bacilos Gram-Positivos/patogenicidade , Periodontite/microbiologia , Biofilmes , Coinfecção/microbiologia , Bacilos Gram-Positivos/fisiologia , Interações Hospedeiro-Patógeno , Humanos , Consórcios Microbianos/fisiologia , Virulência
16.
J Clin Microbiol ; 52(7): 2371-9, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24759706

RESUMO

We evaluated whether the Bruker Biotyper matrix-associated laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) system provides accurate species-level identifications of 147 isolates of aerobically growing Gram-positive rods (GPRs). The bacterial isolates included Nocardia (n = 74), Listeria (n = 39), Kocuria (n = 15), Rhodococcus (n = 10), Gordonia (n = 7), and Tsukamurella (n = 2) species, which had all been identified by conventional methods, molecular methods, or both. In total, 89.7% of Listeria monocytogenes, 80% of Rhodococcus species, 26.7% of Kocuria species, and 14.9% of Nocardia species (n = 11, all N. nova and N. otitidiscaviarum) were correctly identified to the species level (score values, ≥ 2.0). A clustering analysis of spectra generated by the Bruker Biotyper identified six clusters of Nocardia species, i.e., cluster 1 (N. cyriacigeorgica), cluster 2 (N. brasiliensis), cluster 3 (N. farcinica), cluster 4 (N. puris), cluster 5 (N. asiatica), and cluster 6 (N. beijingensis), based on the six peaks generated by ClinProTools with the genetic algorithm, i.e., m/z 2,774.477 (cluster 1), m/z 5,389.792 (cluster 2), m/z 6,505.720 (cluster 3), m/z 5,428.795 (cluster 4), m/z 6,525.326 (cluster 5), and m/z 16,085.216 (cluster 6). Two clusters of L. monocytogenes spectra were also found according to the five peaks, i.e., m/z 5,594.85, m/z 6,184.39, and m/z 11,187.31, for cluster 1 (serotype 1/2a) and m/z 5,601.21 and m/z 11,199.33 for cluster 2 (serotypes 1/2b and 4b). The Bruker Biotyper system was unable to accurately identify Nocardia (except for N. nova and N. otitidiscaviarum), Tsukamurella, or Gordonia species. Continuous expansion of the MALDI-TOF MS databases to include more GPRs is necessary.


Assuntos
Infecções por Actinomycetales/diagnóstico , Actinomycetales/classificação , Técnicas Bacteriológicas/métodos , Listeria/classificação , Listeriose/diagnóstico , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Actinomycetales/química , Actinomycetales/isolamento & purificação , Infecções por Actinomycetales/microbiologia , Bactérias Aeróbias/química , Bactérias Aeróbias/classificação , Bactérias Aeróbias/isolamento & purificação , Análise por Conglomerados , Bacilos Gram-Positivos/química , Bacilos Gram-Positivos/classificação , Bacilos Gram-Positivos/isolamento & purificação , Humanos , Listeria/química , Listeria/isolamento & purificação , Listeriose/microbiologia , Sensibilidade e Especificidade
17.
J Clin Microbiol ; 52(4): 1089-97, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24452159

RESUMO

Reported matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) identification rates of Gram-positive rods (GPR) are low compared to identification rates of Gram-positive cocci. In this study, three sample preparation methods were compared for MALDI-TOF MS identification of 190 well-characterized GPR strains: direct transfer, direct transfer-formic acid preparation, and ethanol-formic acid extraction. Using the interpretation criteria recommended by the manufacturer, identification rates were significantly higher for direct transfer-formic acid preparation and ethanol-formic acid extraction than for direct transfer. Reducing the species cutoff from 2.0 to 1.7 significantly increased species identification rates. In a subsequent prospective study, 215 clinical GPR isolates were analyzed by MALDI-TOF MS, and the results were compared to those for identification using conventional methods, with discrepancies being resolved by 16S rRNA and rpoB gene analysis. Using the direct transfer-formic acid preparation and a species cutoff of 1.7, congruencies on the genus and species levels of 87.4% and 79.1%, respectively, were achieved. In addition, the rate of nonidentified isolates dropped from 12.1% to 5.6% when using an extended database, i.e., the Bruker database amended by reference spectra of the 190 GPR of the retrospective study. Our data demonstrate three ways to improve GPR identification by the Bruker MALDI Biotyper, (i) optimize sample preparation using formic acid, (ii) reduce cutoff scores for species identification, and (iii) expand the database. Based on our results, we suggest an identification algorithm for the clinical laboratory combining MALDI-TOF MS with nucleic acid sequencing.


Assuntos
Algoritmos , Técnicas Bacteriológicas/métodos , Infecções por Bactérias Gram-Positivas/diagnóstico , Bacilos Gram-Positivos/química , Bacilos Gram-Positivos/isolamento & purificação , Manejo de Espécimes/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , RNA Polimerases Dirigidas por DNA/genética , Humanos , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade , Análise de Sequência de DNA
19.
Int J Syst Evol Microbiol ; 63(Pt 9): 3138-42, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23435245

RESUMO

A novel α-amylase/pullulanase-producing bacterium, designated strain GST4(T), was isolated from samples collected from the wastewater of a cassava starch factory in Nanning, Guangxi Autonomous Region, southern China. Cells of strain GST4(T) were rod-shaped bacilli containing ellipsoidal terminal spores and found to be Gram-reaction-positive, aerobic, motile, oxidase-positive, catalase-negative and formed light yellow colonies on agar plates. Strain GST4(T) was able to grow at pH 4.5-8.5 (optimum at pH 5.5), temperatures ranging from 20 to 42 °C (optimum at 37 °C) and salt concentrations of 0-1% (w/v) NaCl (optimum at 0.5%, w/v) on R2A medium. Strain GST4(T) grew heterotrophically on complex carbon substrates and chemolithoautotrophically on inorganic sulfur compounds, as demonstrated by growth on sodium thiosulfate and sulfite as sole electron donors. It can reduce nitrate and nitrite. Strain GST4(T) contained iso-C(15:0) and anteiso-C(15:0) as the major cellular fatty acids and menaquinone 7 (MK-7) as the major respiratory quinone. The cell-wall peptidoglycan was of type A1γ. The genomic DNA G+C content of strain GST4(T) was 53.7 mol%. Physiological and chemotaxonomic characteristics combined with phylogenetic analysis based on 16S rRNA gene sequences revealed that strain GST4(T) was a member of the genus Tumebacillus and most closely related to Tumebacillus permanentifrigoris DSM 18773(T) and Tumebacillus ginsengisoli DSM 18389(T) with 97.3 and 94.5% sequence similarity, respectively. The DNA-DNA relatedness values between strain GST4(T) and T. permanentifrigoris DSM 18773(T), and strain GST4(T) and T. ginsengisoli DSM 18389(T) were 44.0 and 60.4%, respectively. The new isolate differed from those species of the genus Tumebacillus in that it has peritrichous flagella for motility. Based on the evidence obtained from this study, strain GST4(T) represents a novel species of the genus Tumebacillus, for which the name Tumebacillus flagellatus sp. nov. is proposed. The type strain is GST4(T) ( =CGMCC 1.12170(T) =DSM 25748(T)).


Assuntos
Glicosídeo Hidrolases/biossíntese , Bacilos Gram-Positivos/classificação , Filogenia , Águas Residuárias/microbiologia , alfa-Amilases/biossíntese , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/análise , Bacilos Gram-Positivos/genética , Bacilos Gram-Positivos/isolamento & purificação , Manihot , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Peptidoglicano/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/análise
20.
Clin Oral Investig ; 17(4): 1135-42, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-22961460

RESUMO

OBJECTIVE: Information about the spectrum of microorganisms in the intraimplant cavities of two-piece dental implants is scarce. The purpose of this study was to assess the intraimplant microflora of two-piece dental implants by conventional biochemical testing, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and 16 s rDNA gene sequencing. MATERIALS AND METHODS: Ten patients (six men and four women; average age = 66.7 years; age range = 58-78 years) received 35 two-piece titanium implants carrying ball attachments. Biofilm sampling was performed with sterile microbrushes, and nonadherent microbial samples were obtained by injection and reuptake of predefined volumes of NaCl solution. The samples were cultured and analyzed by conventional biochemical testing, MALDI-TOF MS, and 16 s rDNA gene sequencing. RESULTS: Of the 103 species detected, 27 and 33 were identified only in the biofilm and nonadherent microbial samples, respectively. Forty-three species were identified in both types of samples. CONCLUSIONS: Two-piece dental implants harbored a broad spectrum of gram-positive and gram-negative aerobes and anaerobes, especially rods and cocci. CLINICAL RELEVANCE: These findings confirm bacterial translocation from the oral cavity to intraimplant cavities. Microbiological methods as used in this study are necessary to reveal the complete vital microflora of intraimplant cavities.


Assuntos
Projeto do Implante Dentário-Pivô , Implantes Dentários/microbiologia , Microbiota , Alvéolo Dental/microbiologia , Idoso , Bactérias Aeróbias/genética , Bactérias Anaeróbias/genética , Biofilmes , Contagem de Colônia Microbiana , DNA Bacteriano/análise , DNA Bacteriano/genética , Feminino , Bacilos e Cocos Aeróbios Gram-Negativos/genética , Cocos Anaeróbios Gram-Negativos/genética , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/genética , Cocos Gram-Positivos/genética , Bacilos Gram-Positivos/genética , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Sequência de DNA , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
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