Comparison of two MALDI-TOF MS systems for theidentification of clinically relevant anaerobic bacteriain Argentina

Abstract The aim of this study was to compare the performance of two MALDI-TOF MS systems in the identification of clinically relevant strict anaerobic bacteria. The 16S rRNA gene sequencing was the gold standard method when discrepancies or inconsistencies were observed between platforms. A total of 333 isolates were recovered from clinical samples of different centers in Buenos Aires City between 2016 and 2021. The isolates were identified in duplicate using two MALDI-TOF MS systems, BD Bruker Biotyper (Bruker Daltonics, Bremen, Germany) and Vitek MS (bioMèrieux, Marcy-l'Etoile, France). Using the Vitek MS system, the identification of anaerobic isolates yielded the following percentages: 65.5% (n: 218) at the species or species-complex level, 71.2% (n: 237) at the genus level, 29.4% (n: 98) with no identification and 5.1% (n: 17) with misidentification. Using the Bruker Biotyper system, the identification rates were as follows: 85.3% (n: 284) at the species or species-complex level, 89.7% (n: 299) at the genus level, 14.1% (n: 47) with no identification and 0.6% (n: 2) with misidentification. Differences in the performance of both methods were statistically significant (p-values <0.0001). In conclusion, MALDI-TOF MS systems speed up microbial identification and are particularly effective for slow-growing microorganisms, such as anaerobic bacteria, which are difficult to identify by traditional methods. In this study, the Bruker system showed greater accuracy than the Vitek system. In order to be truly effective, it is essential to update the databases of both systems by increasing the number of each main spectrum profile within the platforms.

Resumen El objetivo de este estudio fue comparar el desempeño de dos sistemas MALDI-TOF MS en la identificación de bacterias anaerobias estrictas de interés clínico. La secuenciación del gen 16S ARNr fue el método de referencia utilizado cuando se observaron discrepancias o inconsistencias entre plataformas. Se recuperaron 333 aislados de muestras clínicas de diferentes centros de la Ciudad Autónoma de Buenos Aires entre 2016 y 2021. Los aislados se identificaron por duplicado mediante dos sistemas MALDI-TOF MS: el BD Bruker Biotyper (Bruker Daltonics, Bremen, Alemania) y el Vitek MS (bioMèrieux, Marcy-l'Etoile, Francia). A través del sistema Vitek MS, los mismos fueron identificados a nivel de especie o complejo de especies en un 65,5% (n: 218) y de género en un 71,2% (n: 237), mientras que no se identificaron en un 29,4% (n: 98) y fue incorrecta en el 5,1% (n: 17). Mediante el sistema Bruker Biotyper, dichos valores fueron del 85,3% (n: 284), del 89,7% (n: 299), del 14,1% (n: 47) y del 0,6% (n: 2), respectivamente. La diferencia entre ambos métodos fue estadísticamente significativa (p<0,0001). En conclusión, los sistemas MALDI-TOF MS aceleran la identificación microbiana. Son especialmente útiles para los microorganismos de crecimiento lento, como las bacterias anaerobias, que son difíciles de identificar con los métodos tradicionales. El sistema Bruker demostró ser más preciso que el Vitek MS. Para que estos métodos sean realmente efectivos es fundamental actualizar las bases de datos de ambos sistemas e incrementar el número de espectros de referencia dentro de las plataformas.

Comparison of two MALDI-TOF MS systems for the identification of clinically relevant anaerobic bacteria in Argentina.

The aim of this study was to compare the performance of two MALDI-TOF MS systems in the identification of clinically relevant strict anaerobic bacteria. The 16S rRNA gene sequencing was the gold standard method when discrepancies or inconsistencies were observed between platforms. A total of 333 isolates were recovered from clinical samples of different centers in Buenos Aires City between 2016 and 2021. The isolates were identified in duplicate using two MALDI-TOF MS systems, BD Bruker Biotyper (Bruker Daltonics, Bremen, Germany) and Vitek MS (bioMèrieux, Marcy-l'Etoile, France). Using the Vitek MS system, the identification of anaerobic isolates yielded the following percentages: 65.5% (n: 218) at the species or species-complex level, 71.2% (n: 237) at the genus level, 29.4% (n: 98) with no identification and 5.1% (n: 17) with misidentification. Using the Bruker Biotyper system, the identification rates were as follows: 85.3% (n: 284) at the species or species-complex level, 89.7% (n: 299) at the genus level, 14.1% (n: 47) with no identification and 0.6% (n: 2) with misidentification. Differences in the performance of both methods were statistically significant (p-values <0.0001). In conclusion, MALDI-TOF MS systems speed up microbial identification and are particularly effective for slow-growing microorganisms, such as anaerobic bacteria, which are difficult to identify by traditional methods. In this study, the Bruker system showed greater accuracy than the Vitek system. In order to be truly effective, it is essential to update the databases of both systems by increasing the number of each main spectrum profile within the platforms.

Brain abscesses caused by anaerobic microorganisms: a systematic review

The objective of this study was to perform a systematic review of the characteristics, causative microorganisms and outcome of brain abscesses caused by anaerobic bacteria over the past 25 years. We reviewed studies on brain abscesses which included infection due to anaerobic microorganisms published between 1998 and 2022. We excluded reports with polymicrobial infections (more than 2 anaerobic bacteria isolated) and those that do not provide enough information to make comparisons, the reports with only one case of brain abscess due to anaerobes, as well as those focused on an only anaerobic bacterium. Also, we have excluded the cases in pediatric population. We searched the scientific literature through the Cochrane Library, EMBASE and PubMed/MEDLINE databases for studies of this condition. We finally included 28 studies with 6,167 patients, of which 715 (11.5%) were cases caused by anaerobic bacteria. There was a male predominance (70%) and mean age of 40.3 years. Most infections were monomicrobial (59.4%). The most common anaerobic microorganisms isolated were Bacteroides spp (43.4%) and Gram-positive anaerobic cocci (35.1%). Cases of brain abscesses caused by anaerobic bacteria were most frequent in Asia and Europe. The source of infection most frequent was otogenic in 84.6% followed by a neurosurgery procedure infection in 23% of patients. The main symptom observed was headache in 95.6% of patients followed by fever (69.5%). Surgical treatment was performed in 48 % of patients and the percentage of patients in whom antibiotic treatment was applied range 88.8% to 100%. The main limitation of this review is the non-inclusion of studies published before of 1998 in which MALDI-TOF MS system had not been introduced in the majority of laboratories for routine identification. (AU)

El objetivo de este estudio fue realizar una revisión sistemática de las características, los microorganismos causantes y evolución de los abscesos cerebrales causados por bacterias anaerobias en los últimos 25 años. Revisamos los estudios sobre abscesos cerebrales que incluyeron infección por microorganismos anaerobios publicados entre 1998 y 2022. Se excluyeron estudios con infecciones polimicrobianas (más de 2 bacterias anaerobias aisladas) y aquellos que no proporcionaban suficiente información para realizar comparaciones, los estudios con solo un caso de absceso por anaerobios así como aquellos focalizados solo en una bacteria anaerobia. También se excluyeron los casos en población pediátrica. Se realizó búsqueda de la literatura científica a través de la librería Cochrane y base de datos EMBASE y PubMed/MEDLINE para estudios con esas características. Se incluyeron finalmente 28 estudios con 6167 pacientes, de los cuales 715 (11,5%) fueron casos causados por bacterias anaerobias. Hubo predominio masculino (70%) y edad media de 40,3 años. La mayoría de las infecciones fueron monomicrobianas (59,4%). Los microorganismos anaerobios más comunes aislados fueron Bacteroides spp (43,4%) y cocos anaerobios grampositivos (35,1%). Los casos de abscesos cerebrales causados por bacterias anaerobias fueron más frecuentes en Asia y Europa. La fuente de infección más frecuente fue la otogénica en un 84,6% seguida de una infección por procedimiento de neurocirugía en un 23% de los pacientes. El principal síntoma observado fue la cefalea en el 95,6% de los pacientes seguido de fiebre (69,5%). Se realizó tratamiento quirúrgico en el 48 % de los pacientes y el porcentaje de pacientes en los que se aplicó tratamiento antibiótico oscila entre el 88,8 % y el 100 %. La principal limitación de esta revisión fue la no inclusión de estudios anteriores a 1998 en los que todavía no se había introducido MALDI-TOF MS en la mayoría de los laboratorios para el diagnóstico rutinario. (AU)

Empiema pleural por Parvimonas micra en un paciente inmunocompetente: reporte de un caso / Pleural empyema by Parvimonas micra in an immunocompetent patient: a case report

Se presenta el caso de un paciente joven inmunocompetente, con antecedente de tuberculosis pulmonar, que acude al hospital por un cuadro clínico subagudo de fiebre persistente, baja de peso, disnea y abolición del murmullo vesicular. La tomografía de tórax mostró un extenso empiema en hemitórax izquierdo. Se le toman muestras para detección de gérmenes comunes y se le colocan un tubo de drenaje torácico y se inicia antibioticoterapia. La prueba de MALDI-TOF MS identificó a Parvimonas micra, una bacteria anaerobia, comensal de la flora oral, asociado a periodontitis severa, escasamente reportado en empiema pleural, especialmente, en personas inmunocompetentes. En la evaluación odontológica se realizó el diagnóstico de gingivitis y pericoronaritis de la tercera molar. El paciente evolucionó favorablemente. Se sugiere que, en casos de empiemas pleurales subagudos o crónicos, se debe considerar, además de las micobacterias, como agente etiológico al Parvimonas micra, y optar por exámenes como MALDI-TOF MS o secuenciamiento del 16S rRNA, colocación de tubo de tórax, cobertura antibiótica empírica y evaluación odontológica.

We present the case of a young immunocompetent patient, with a history of pulmonary tuberculosis, who attended the hospital with a subacute clinical picture of persistent fever, weight loss, dyspnea and abolition of vesicular murmur. Chest CT scan showed an extensive empyema in the left hemithorax. Samples were taken for detection of common germs. Then, a chest drainage tube was placed and antibiotic therapy started. The MALDI-TOF MS test identified Parvimonas micra, an anaerobic bacterium, commensal to the oral flora, associated with severe periodontitis, but rarely reported in cases of pleural empyema, especially in immunocompetent patients. Gingivitis and pericoronaritis of the third molar were diagnosed during oral evaluation. The patient progressed favorably. Parvimonas micra should be considered as a possible etiological agent in cases of subacute or chronic pleural empyema, in addition to mycobacteria. Tests such as MALDI-TOF MS or 16S rRNA sequencing, chest tube placement, empirical antibiotic coverage and an adequate oral evaluation should be considered in these cases.

Diagnostic and therapeutic approaches in deep neck infections: an analysis of 74 consecutive patients / Abordagens diagnósticas e terapêuticas em infecções cervicais profundas: uma análise de 74 pacientes consecutivos

Abstract Introductıon: Deep neck infections are a group of diseases with serious complications and mortality, which can occur as a result of common diseases in the community and which have surgical and medical treatment options. Objectives: Patients ages, genders, complaints, physical examination findings, hospitalization complaints, history of antibiotic use before the application, additional diseases, radiological tests and analysis of examinations, type of treatment method, antibiotic agents selected in treatment, bacterial culture results, duration of hospitalization, complications, mortality rates were systematically recorded. In the study, anaerobic bacterial factors, which are difficult to produce in routine, were produced by considering special transport conditions and culture media. Methods: A total of 74 patients who were hospitalized in the Department of Otorhinolaryngology, University of Mersin, between 01.07.2016 and 01.07.2017 for deep neck infection were evaluated prospectively. The study included 37 female and 37 male patients. The ages of the patients ranged from 1 to 69 and the mean age was 31 years. Results: According to the analysis of the obtained data, there was a statistically significant relationship between the patients with additional diseases and the treatment modalities of the patients (p = 0.017). The surgical treatment rate was increased in this group of patients. In patients with a history of antibiotic use, it was found that patients in the pediatric group were in hospital longer in terms of length of stay compared to adults (p = 0.036). In adult patients who underwent surgery, the absorptive long axis was found to be longer in mm than in patients receiving isolated medical treatment (p = 0.008). Conclusions: Deep neck infections is a disease group that seriously concerns public health, with significant mortality and morbidity. Ensuring airway safety of patients should be the first intervention. Abscesses located lateral to the tonsil capsule may not drain adequately without concomitant tonsillectomy.

Resumo Introdução: As infecções cervicais profundas constituem um grupo de doenças com graves complicações e mortalidade, que podem ocorrer como resultado de doenças comuns na comunidade e que têm opções de tratamento médico e cirúrgico. Objetivo: Detectar bactérias anaeróbias e comparar a eficácia do tratamento médico-cirúrgico em diferentes faixas etárias. Método: Foram sistematicamente registrados idade dos pacientes, sexo, queixas, achados do exame físico, queixas na hospitalização, histórico de uso de antibióticos, doenças adicionais, exames radiológicos e análise dos exames, tipo de tratamento, agentes antibióticos selecionados no tratamento, resultados de cultura bacteriana, duração da hospitalização, complicações e taxas de mortalidade. No estudo, culturas bacterianas anaeróbias, difíceis de obter rotineiramente, mesmo considerando-se condições especiais de transporte e meios, foram obtidas. Foram avaliados prospectivamente 74 pacientes internados no Departamento de Otorrinolaringologia da Universidade de Mersin, entre 01.07.2016 e 07.07.2017, devido a infecção cervical profunda. O estudo incluiu 37 pacientes do sexo feminino e 37 do masculino. A idade dos pacientes variou de 1 a 69 anos e a média foi de 31. Resultados: De acordo com a análise dos dados obtidos, houve uma relação estatisticamente significante entre os pacientes com doenças adicionais e as modalidades de tratamento dos pacientes (p = 0,017). A taxa de tratamento cirúrgico foi maior nesse grupo de pacientes. Em pacientes com histórico de uso de antibióticos, verificou-se que aqueles do grupo pediátrico permaneceram mais tempo hospitalizados em comparação aos adultos (p = 0,036). Nos pacientes adultos submetidos à cirurgia, verificou-se que o eixo longo absortivo era mais longo em mm do que nos pacientes que receberam tratamento médico isolado (p = 0,008). Conclusão: As infecções cervicais profundas constituem um grupo de doenças que são preocupantes na saúde pública, com sua mortalidade e morbidade. Garantir a segurança das vias aéreas dos pacientes deve ser a primeira intervenção. Os abscessos localizados lateralmente à cápsula tonsilar podem não apresentar drenagem sem a tonsilectomia.

Use of wide-field optical fluorescence for visualization of oral biofilm in a patient with peri-implant mucositis: a new approach / Uso da fluorescência óptica de campo amplo para visualização de biofilme oral em paciente com mucosite peri-implantar: uma nova abordagem

ABSTRACT Peri-implant diseases, caused by bacteria from biofilm related to dental implants, are one of the main causes of late loss of implants. In this sense, peri-implant diseases are divided into peri-implant mucositis, when it affects only the soft tissues, and peri-implantitis, when there is a bone involvement, which can lead to the failure of dental implant therapy. Thus, biofilm removal is essential for peri-implant health, allowing long-term success in implant therapy. To improve the visualization of oral biofilm, which is usually transparent or colorless, disclosing agents have been routinely used. However, disclosing agents have allergenic potential and can cause staining extrinsically in restorative and prosthetic materials, leading to aesthetic impairment. Thus, the use of fluorescence has been studied as an alternative for visualization of oral biofilm. Therefore, this report describes the use of wide-field optical fluorescence for visualization of oral biofilm associated with implants and teeth, in a routine appointment and follow-up of a partially edentulous patient with peri-implant mucositis. In addition, this report showed wide-field optical fluorescence can be used in a clinical routine of care of patients with dental implants. In this sense, wide-field optical fluorescence allowed easy and immediate visualization of the mature oral biofilm for its adequate removal, evaluation of the quality of restoration to sealing of screw access-hole of implant and identification of cariogenic lesions, without risk of allergic reactions or staining of prostheses and restorations.

RESUMO Doenças peri-implantares, causadas por bactérias de biofilme relacionadas a implantes dentários, são uma das principais causas de perda tardia de implantes. Nesse sentido, as doenças peri-implantares são divididas em mucosite peri-implantar, quando afeta apenas tecidos moles, e peri-implantite, quando há comprometimento ósseo, o que pode levar ao fracasso da terapia com implantes dentários. Assim, a remoção do biofilme é essencial para a saúde peri-implantar, permitindo sucesso a longo prazo na terapia com implantes. A fim de melhorar a visualização do biofilme oral, que geralmente é transparente ou incolor, agentes reveladores têm sido rotineiramente utilizados. No entanto, esses agentes têm potencial alergênico e podem causar manchas extrinsecamente em materiais restauradores e protéticos, levando a prejuízo estético. Assim, o uso da fluorescência tem sido estudado como alternativa para visualização do biofilme oral. Este relato descreve o uso da fluorescência óptica de campo amplo para visualização do biofilme oral associado a implantes e dentes em uma consulta de acompanhamento de rotina de uma paciente parcialmente edêntula com mucosite peri-implantar. Além disso, este relato evidenciou que a fluorescência óptica de campo amplo pode ser utilizada dentro da rotina clínica de atendimento de pacientes com implantes dentários. Nesse sentido, a fluorescência óptica de campo amplo permitiu a visualização fácil e imediata do biofilme oral maduro para sua remoção adequada, a avaliação da qualidade da restauração do selamento do orifício de acesso do parafuso do implante e a identificação de lesões cariogênicas, sem risco de reações alérgicas ou manchamento de próteses e restaurações.

Anal abscess microbiology as an anal fistula predictor / Microbiologia de abscesso anal como preditivo de fístula anal

ABSTRACT In order to determine the microbiology of anal abscess as a predictor of anal fistulas in patients who attended the external consultation of the Coloproctology unit of the Dr. Antonio María Pineda University Central Hospital, a prospective, descriptive, longitudinal study was conducted from September 2018 to July 2019. In this study, the population consisted of patients with a diagnosis of anal abscess, without associated comorbidities or contraindications for surgery, who agreed to be included in the study. A non-probabilistic, intentional sample consisting of 42 patients was determined. An appointment-based study protocol was applied by outpatient for patients who met the inclusion criteria applied, to perform due medical history through anamnesis, physical examination and culture taking of suppuration from the anal abscess to subsequently establish medical and surgical behavior thereof. The results were expressed in absolute numbers and percentages, a prevalence of ischiorectal abscesses was observed, followed by deep post-anal space abscesses. Anaerobic bacteria were isolated in 100% of the sample. In all fistulized patients, E. Coli was isolated as a predominant germ.

RESUMO Um estudo prospectivo, descritivo e longitudinal foi realizado de setembro de 2018 a Julho 2019 para determinar a microbiologia do abscesso anal como preditivo de fístulas anais em pacientes que compareceram à consulta externa da unidade de Coloproctologia do Hospital Central da Universidade Dr. Antonio María Pineda. Neste estudo, a população foi composta por pacientes com diagnóstico de abscesso anal, sem comorbidades ou contraindicações associadas à cirurgia, que concordaram em participar do estudo. Uma amostra intencional não probabilística, composta por 42 pacientes foi determinada. Um protocolo de estudo com base na consulta em regime ambulatorial foi aplicado aos pacientes que atenderam os critérios de inclusão estabelecidos, para realizar a anamnese, o exame físico e a devida cultura da supuração do abscesso anal para posteriormente estabelecer o comportamento médico e cirúrgico. Os resultados foram expressos em números absolutos e porcentagens, observando-se a prevalência de abscessos isquiorretais, seguidos por abscessos profundos no espaço pós-anal. Bactérias anaeróbias foram isoladas em 100% das amostras. Em todos os pacientes com fístulas, E. Coli foi isolada como um germe predominante.

Recuperación de bacterias aerobias y anaerobias de pacientes con apendicitis aguda mediante botellas de hemocultivo / Recovery of aerobic and anaerobic bacteria from patients with acute appendicitis using blood culture bottles

Introducción. La apendicitis aguda es la primera causa de abdomen agudo; sin embargo, poco se conoce sobre las bacterias asociadas y su perfil de sensibilidad. Objetivo. Identificar y determinar el patrón de resistencia de las bacterias aerobias y anaerobias aisladas en cultivo de líquido periapendicular tomado de los pacientes con apendicitis aguda, y establecer la proporción de bacterias según la fase clínica. Materiales y métodos. Se llevó a cabo un estudio descriptivo y prospectivo en el Hospital Universitario de San José de Bogotá (Colombia), en pacientes mayores de 16 años sometidos a apendicectomía abierta. Se tomaron muestras de líquido periapendicular, las cuales se sembraron directamente en botellas de hemocultivos para aerobios y anaerobios. Resultados. Se incluyeron 154 pacientes. Del total de cultivos, el 87 % (n=134) fueron positivos: 77 % (n=118) para aerobios y 51 % (n=79) para anaerobios. La proporción de cultivos positivos fue inferior en los casos de apendicitis no complicada, en comparación con aquellos de apendicitis complicada (80 % (66/83) Vs. 95 % (67/71); p=0,003). Los microorganismos aislados con mayor frecuencia fueron: Escherichia coli (53 %) (n=84), Bacteroides sp. (25 %) (n=25), Propionibacterium acnes (21 %) (n=21), Staphylococci coagulasa negativo (17 %) (n=27), Enterococcus sp. (10 %) (n=15) y Fusobacterium sp. (11 %) (n=11). La sensibilidad de E. coli a la amplicilina sulbactam fue de 30 %. La sensibilidad de Bacteroides spp. a la clindamicina y la ampicilina sulbactam fue de 91 %. El 100 % de los anaerobios fueron sensibles a piperacilina tazobactam, ertapenem, meropenem y metronidazol. Conclusiones. Los cultivos intraoperatorios son pertinentes en la apendicitis para determinar el patrón epidemiológico local, y establecer los antibióticos profilácticos y terapéuticos para esta enfermedad. Su siembra directa en botellas de hemocultivo permite una gran recuperación de microorganismos.

Introduction: Acute appendicitis is the first cause of acute abdomen, however, there is a little information about the associated bacteria and its sensibility profile. Objetive: To identify and to determine the resistance pattern of aerobic and anaerobic bacteria isolated in periapendicular fluid cultures taken in patients with acute appendicitis and to establish the proportions of isolates according to the clinical phase. Materials and methods: A descriptive and prospective study was undertaken at the Hospital Universitario de San José (Bogotá, Colombia) of patients older than sixteen years of age, undergoing an open appendectomy. A sample of periappendiceal fluid was taken, which was deposited directly into aerobic and anaerobic blood culture bottles. Results: One hundred and fifty-four patients were included. The overall positivity of cultures was 87% (n=1344); 77% (n=118) for aerobes and 51% (n=79) for anaerobes. The proportion of positive cultures was lower in the uncomplicated appendicitis cases as compared to the complicated ones (80% (66/83) vs. 95%(67/71), p = 0.003). The microorganisms isolated most frequently were: Escherichia coli (53%) (n=84); Bacteroides spp. (25%) (n=25); Propionibacterium acnes (21%) (n=21); coagulase negative Staphylococci (17%) (n=27); Enterococcus spp. (11%) (n=15), and Fusobacterium spp. (11%) (n=11). The sensitivity of E.coli to ampicillin/sulbactam was 30%. The sensitivity of Bacteroides spp. to clindamycin and ampicillin/sulbactam was 91%. All anaerobe isolates were sensitive to piperacillin/tazobactam, ertapenem, meropenem and metronidazole. Conclusions: Intraoperative cultures in acute appendicits are relevant in order to determine the local epidemiological pattern and to establish prophylactic and therapeutic antibiotics for this pathology; direct inoculation in blood culture bottles allows a high recovery of microorganisms.

Evaluation of MALDI-TOF mass spectrometry (VITEK-MS) compared to the ANC card (VITEK 2) for the identification of clinically significant anaerobes / Avaliação da espectrometria de massas MALDI-TOF (VITEK-MS) diante do cartão ANC (VITEK 2) na identificação de anaeróbios clinicamente significantes

ABSTRACT Introduction: The identification of anaerobic bacteria by conventional methods employed in clinical laboratories requires a lot of work and a long response time [turnaround time (TAT)]. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is an accurate, rapid and inexpensive technique with promising results for bacterial identification. Objective: To evaluate the MALDI-TOF mass spectrometry (VITEK-MS, bioMérieux, France) compared to the ANC card (VITEK 2, bioMérieux, France) for the identification of anaerobes, and also veriying the cost variation between both methodologies. Methods: 421 anaerobes were concomitantly identified by ANC (VITEK 2) and MALDI-TOF (VITEK MS). The conflicting results or those presenting low differentiation of the species were subjected to the 16S ribosomal ribonucleic acid (rRNA) sequencing. Results: Thirty-five strains were not identified by ANC (VITEK 2), and only one isolate was not identified by MALDI-TOF (VITEK MS). From the 386 anaerobes identified by the two methodologies, 97% agreement was observed on the identification of genus and species between the methodologies. Thirteen (3%) isolates were submitted to 16S sequencing. The agreement observed was 70% using ANC (VITEK 2) using 92% by MALDI-TOF (VITEK MS). Conclusion: Both methodologies showed an excellent performance for the identification of the strains tested with great differences in relation to cost-benefit. MALDI-TOF MS allowed 35 additional identifications and a saving of BRL$ 7,786 with the release of culture positive result five days ahead of the ANC (VITEK 2). TAT reduction may contribute to a successful clinical resolution.

RESUMO Introdução: A identificação das bactérias anaeróbias por métodos convencionais empregados nos laboratórios clínicos demanda muito trabalho e um longo tempo de resposta (TAT). A espectrometria de massa por ionização e dessorção a laser assistida por matriz (MALDI-TOF MS) é uma técnica precisa, rápida e barata, com resultados promissores para a identificação bacteriana. Objetivo: Avaliar a espectrometria de massas MALDI-TOF (VITEK MS, bioMérieux, France) diante do cartão ANC (VITEK 2, bioMérieux, France) para a identificação de anaeróbios, bem como verificar a variação de custos entre as metodologias. Métodos: Foram identificados 421 anaeróbios concomitantemente pelo ANC (VITEK 2) e pelo MALDI-TOF (VITEK MS). Os resultados discordantes ou que apresentaram baixa discriminação das espécies foram submetidos ao sequenciamento do 16S do ácido ribonucleico ribossonal (rRNA). Resultados: Trinta e cinco cepas não foram identificadas pelo ANC (VITEK 2), e somente um isolado ficou sem identificação pelo MALDI-TOF (VITEK MS). Dos 386 anaeróbios identificados pelas duas metodologias, a concordância na identificação de gênero e espécie foi observada em 97%. Treze (3%) isolados foram submetidos ao sequenciamento do 16S; a concordância observada foi de 70% com o ANC (VITEK 2) e 92% com MALDI-TOF (VITEK MS). Conclusão: Ambas as metodologias demonstraram ótimo desempenho para identificação das cepas testadas com grandes diferenças em relação ao custo-benefício. O MALDI-TOF MS permitiu 35 identificações adicionais e uma economia de R$ 7.786,00 com a liberação do resultado positivo da cultura cinco dias à frente do ANC (VITEK 2). A redução do TAT pode contribuir para uma resolução clínica bem-sucedida.

Atividade antimicrobiana, anti-inflamatória, citotoxicidade e genotoxicidade do extrato glicólico de Stryphnodendron barbatiman (Vell.) Mart. (Barbatimão) / Antimicrobial, anti-inflammatory, cytotoxicity and genotoxicity activity of the glycolic extract of Stryphnodendron barbatiman (Vell. ) Mart

Stryphnodendron barbatiman (Vell.) Mart (barbatimão) tem sido utilizado popularmente como anti-inflamatório e antimicrobiano. Este estudo se propôs a realizar a análise fitoquímica do extrato de S. barbatiman (Vell.) Mart. (barbatimão) e verificar in vitro algumas de suas atividades biológicas, como: antimicrobiana, sobre culturas planctônicas e biofilmes de Fusobacterium nucleatum, Porphyromonas endodontalis, Porphyromonas gingivalis, e Parvimonas micra; citotoxicidade e genotoxicidade em macrófagos murinos (RAW 264.7), fibroblastos murinos (L929) e queratinócitos humanos (HACAT); e ação anti-inflamatória sobre RAW 264.7 estimuladas por lipopolissacarídeo (LPS) de Escherichia coli. A atividade antimicrobiana sobre culturas planctônicas foi avaliada pelo método de microdiluição em caldo. Após, as concentrações efetivas foram avaliadas sobre biofilmes monomicrobianos. A análise de citotoxicidade foi verificada pelo teste de redução do brometo de 3(4,5-dimetiltiazol-2-yl)2,5-difeniltetrazólio (MTT) e a de genotoxicidade pelo teste de micronúcleos. A atividade anti-inflamatória foi analisada quantificando os níveis de citocinas pró-inflamatórias (TNF-α, IL-1ß, IL-6 e IL-17) e anti-inflamatória (IL-10), por enzyme-linked immunosorbent assay (ELISA). Os dados foram analisados por ANOVA e Teste de Tukey, ou Kruskal-Wallis e teste de Dunns, com nível de significância de 5% (P≤0,05). O extrato foi eficaz sobre os biofilmes, redução entre 54 e 100%. O extrato não foi citotóxico para RAW 264.7 e L929 no tempo de 24 h nem para HACAT por 5 min e em 24 h 50 e 100 mg/mL aumentaram a viabilidade celular. O extrato de S. barbatiman (Vell.) Mart. não foi genotóxico para RAW 264.7. O extrato associado ao LPS aumentou a produção de todas as citocinas estudadas, exceto do TNF-α. Concluiu-se que o extrato de barbatimão foi eficaz sobre biofilme mono espécie de bactérias anaeróbias em concentrações não citotóxicas para RAW 264.7, L929 e HACAT (AU)

Anti-inflammatory and antimicrobial are popular uses of Stryphnodendron adstringens (Mart.) Coville (barbatimão). This study proposed to perform the phytochemical analysis of S. barbatiman (Vell.) Mart. (barbatimão) extract and to verify in vitro some of its biological activities as antimicrobial on planktonic cultures and biofilms of Fusobacterium nucleatum, Porphyromonas endodontalis, Porphyromonas gingivalis, and Parvimonas micra; cytotoxicity and genotoxicity on human keratinocytes (HACAT), murine macrophages (RAW 264.7) and fibroblasts (L929), and anti-inflammatory action on RAW 264.7. The antimicrobial activity on planktonic cultures was evaluated by broth microdilution. Afterwards, the effective concentrations were evaluated on mono microbial biofilms. The cytotoxicity analysis was verified by the MTT test and the genotoxicity by the micronucleus test. Antiinflammatory activity was evaluated in RAW 264.7 stimulated by lipopolysaccharide (LPS) of Escherichia coli and treated with the extract, quantifying the levels of proinflammatory cytokines (TNF-α, IL-1ß, IL-6 and IL-17) and anti-inflammatory (IL-10), by enzyme-linked immunosorbent assay (ELISA). The data that presented normal distribution were analyzed by ANOVA and Tukey's test, and those that were not, were analyzed by Kruskal-Wallis and Dunns test, with a significance level of 5% (p≤0.05). Barbatimão extract was effective on biofilms, reduction between 54 and 100%. The extract was not cytotoxic to RAW 264.7 and L929 in the 24 hr time nor to HACAT for 5 min and in 24 h 50 and 100 mg / ml increased cell viability. The extract of S. barbatiman (Vell.) Mart. was not genotoxic for RAW 264.7. The LPS-associated extract increased the production of all cytokines studied except for TNF-α. In conclusion, barbatimão extract was effective on mono-species biofilm of anaerobic bacteria at non-cytotoxic concentrations for RAW 264.7, L929 and HACAT (AU)

Ação antimicrobiana e citotoxicidade de extratos aquoso e glicólico de própolis sobre bactérias anaeróbias de importância odontológica / Antimicrobial action and cytotoxicity of aqueous and glycolic extracts of propolis on anaerobic bacteria of dental importance

As plantas medicinais e os fitoterápicos têm sido utilizados como coadjuvantes e alternativos no combate a diversas doenças com crescente frequência, no entanto, na Odontologia seu uso ainda é bastante limitado. Com isso, o objetivo deste estudo é avaliar a ação antimicrobiana dos extratos aquoso e glicólico de própolis verde sobre micro-organismos anaeróbios de interesse odontológico, bem como sua citotoxicidade, a fim de introduzir e incentivar o uso efetivo e sistemático desse fitoterápico em produtos como dentifrícios e enxaguatórios bucais no combate a cáries e doenças periodontais. Os extratos comerciais aquoso e glicólico de própolis foram obtidos das empresas Apis Flora e Mapric, respectivamente. Para avaliação da atividade antimicrobiana foram utilizadas cepas-padrão (ATCC) de Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis Porphyromonas gingivalis e Prevotella intermedia em cultura planctônica, verificando a concentração inibitória mínima e concentração microbicida mínima (CIM e CMM), segundo Clinical and Laboratory Standards Institute. Para biofilmes monotípicos, suspensões padronizadas (107 céls/mL) foram adicionadas em poços de microplacas e após 48 h em anaerobiose foram tratados com 3 concentrações do extrato de própolis (n=12) por 5 min. Foi incluído um controle positivo (solução fisiológica) e um controle negativo (clorexidina). O biofilme foi mensurado pelos testes MTT e Cristal Violeta. Para análise de citotoxicidade, queratinócitos humanos (HaCaT) foram cultivados e colocados em contato com os extratos por 5 min e 24h. Os dados foram analisados estatisticamente pelos testes ANOVA e Tukey Test (5%). Os resultados mostraram que os extratos tiveram ação antimicrobiana contra as suspensões planctônicas e os biofilmes monotípicos dos patógenos, sendo tão ou mais eficazes que a clorexidina. Quanto à citotoxicidade, observou-se diminuição da viabilidade celular dos queratinócitos humanos após a aplicação dos extratos, do mesmo modo que a clorexidina, sendo o extrato glicólico menos citotóxico que o aquoso. Com isto conclui-se que os extratos comerciais aquoso e glicólico de própolis verde tem ação antimicrobiana contra os micro-organismos anaeróbios orais estudados e apresentam potencial para serem utilizados nos tratamentos contra os referidos patógenos, já que no que se refere à citotoxicidade, se comportaram de forma semelhante à Clorexidina, cujo uso é conhecidamente seguro e eficaz(AU)

Medicinal plants and herbal medicines have been used as adjuvants and alternatives in fighting various diseases with increasing frequency. However, in dentistry its use is still quite limited. Therefore, the objective of this study is to evaluate the antimicrobial action of the aqueous and glycolic extracts of green propolis on anaerobic microorganisms of dental interest, in order to introduce and encourage the effective and systematic use of this herbal medicine in products such as dentifrices and mouthwashes in the fight against tooth decay and periodontal diseases. Aqueous and glycolic commercial extracts of propolis were obtained from Apis Flora and Mapric companies, respectively.To evaluate the antimicrobial activity, standard strains (ATCC) of Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in planktonic culture was used, verifying the minimum inhibitory concentration and minimum microbicide concentration (MIC and CMM), according to Clinical and Laboratory Standards Institute. For monotypic biofilms, standardized suspensions (107 cells / mL) was added to microplate wells and after 48 h in anaerobiosis was treated with 3 concentrations of propolis extracts (n = 12) for 5 min. A positive control (saline solution) and a negative control (chlorhexidine) was included. The biofilm was measured by the MTT and Violet Crystal tests. For cytotoxicity analysis, human keratinocytes (HaCaT) were cultured and placed in contact with the extracts for 5 min and 24 h. Data were analyzed statistically by ANOVA and Tukey Test (5%). The results showed that the extracts had antimicrobial action against planktonic suspensions and monotypic pathogen biofilms, being as or more effective than chlorhexidine. As for cytotoxicity, the cellular viability of human keratinocytes was observed after application of the extracts, in the same way as chlorhexidine, the glycolic extract being less cytotoxic than aqueous. It is concluded that the aqueous and glycolic extracts of green propolis have an antimicrobial action against the studied oral anaerobic microorganisms and and have the potential to be used in the treatments against these pathogens, since in cytotoxicity they behaved in a way similar to chlorhexidine, the use of which is known to be safe and effective(AU)

Potencial antimicrobiano de extratos glicólicos vegetais sobre bactérias anaeróbias / Antimicrobial potential of plant glycolic extracts on anaerobic bacteria

A resistência microbiana aos antibióticos disponíveis é preocupação constante, devido à dificuldade no tratamento de infecções causadas por cepas resistentes, em decorrência do uso indiscriminado de antimicrobianos. Assim, a busca por terapias antimicrobianas alternativas tem sido crescente e necessária, sendo a fitoterapia umas das opções de escolha. O objetivo do presente estudo foi analisar a atividade antibacteriana de extratos glicólicos de Achyrocline satureioides (macela), Cynara scolymus (alcachofra), Hamamelis virginiana (hamamelis) e Persea americana (abacateiro), pelos períodos de 5 min e 24 h de exposição sobre bactérias anaeróbias Fusobacterium nucleatum subsp. nucleatum, Parvimonas micra, Porphyromonas endodontalis e Porphyromonas gingivalis, em culturas planctônica e biofilmes. As bactérias armazenadas a -80ºC foram ativadas em caldo Brucella enriquecido (hemina 1%, menadiona 1% e sangue de carneiro desfibrinado 5%) e incubadas em câmara de anaerobiose por 48 h a 37ºC por sete dias. A partir de culturas puras, o teste de microdiluição em caldo foi conduzido em microplacas por meio de suspensões bacterianas padronizadas em solução fisiológica estéril (NaCl 0,9%) e diluições dos extratos em caldo, sendo as placas incubadas por 48 h a 37ºC em anaerobiose. Alíquotas de cada poço foram semeadas em ágar Brucella enriquecido. Após incubação, a Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM) foram determinadas. As concentrações efetivas de cada extrato foram aplicadas sobre os biofilmes de cada espécie, formados em microplacas a partir de suspensões bacterianas puras padronizadas na escala 0,5 de McFarland. As microplacas foram incubadas por sete dias a 37ºC para formação dos biofilmes, sendo o meio trocado a cada 48 h. Os biofilmes foram tratados por 5 min e 24 h. Em seguida, foram lavados e desprendidos por homogeneizador ultrassônico. As suspensões diluídas foram adicionadas em ágar Brucella enriquecido. Após 48 h, as Unidades Formadoras de Colônia por mililitro (UFC/mL) foram determinadas. Os resultados foram analisados por ANOVA e teste de Tukey ou por Kruskal-Wallis e teste Dunns, ambos com nível de significância de 5% (p≤0,05). Sobre as culturas planctônicas, a CIM e CBM dos extratos foi determinada apenas para F. nucleatum. A CBM dos extratos de A. satureioides, C. scolymus e P. americana foi obtida sobre P. micra. Não foi obtida atividade bactericida para P. endodontalis e P. gingivalis. Sobre biofilmes, todas as espécies apresentaram reduções significativas quando expostas aos extratos em ambos os tempos. Pode-se concluir que os extratos testados apresentaram efeito bacteriostático sobre F. nucleatum. Atividade bactericida dos extratos foi observada sobre F. nucleatum, bem como sobre P. micra, exceto para H. virginiana. Os extratos avaliados também apresentaram efeito antibiofilme sobre F. nucleatum, P. micra, P. endodontalis e P. gingivalis por 5 min e 24 h de exposição(AU)

Microbial resistance to antibiotics available is constant concern, due to the difficulty in treating infections caused by resistant strains as a result of the indiscriminate use of antimicrobials. Thus, the search for antimicrobial alternative therapies has been growing and necessary, being one option the herbal medicine. The objective of the present study was to analyze the antibacterial activity to Achyrocline satureioides glycolic extracts (macela), Cynara scolymus (artichoke), Hamamelis virginiana (Witch-Hazel) and Persea americana (avocado), for periods of 5 min and 24 h from exhibition on anaerobic bacteria Fusobacterium nucleatum subsp. nucleatum, Parvimonas micra, Porphyromonas gingivalis and Porphyromonas endodontalis in planktonic communities and biofilms. Bacteria stored at -80°C have been activated in Brucella broth enriched (hemin 1%, menadione 1% and defibrinated sheep blood 5%) and incubated in anaerobiose chamber for 48 h at 37° C for seven days. From pure cultures, the microdiluição test in broth was conducted in microplates through standardized bacterial suspensions in sterile saline solution (NaCl 0.9%) and dilution of the extracts in broth, being incubated plates for 48 h at 37° C in anaerobiosis. Aliquots of each well were sown in Brucella agar enriched. After incubation, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined. Effective concentrations of each extract were applied on the biofilms of each species, formed in microplates from pure bacterial suspensions in 0.5 McFarland scale standard. The microplates were incubated for 7 days at 37°C for the formation of biofilms, being the culture medium replaced every 48 h. Biofilms were treated for 5 min and 24 h have been washed and given off by ultrasonic homogenizer. Dilute suspensions were added in Brucella agar enriched. After 48 h, the Colony Forming Units per milliliter (CFU/ml) were determined. The results were analyzed by ANOVA and Tukey test, or Kruskal-Wallis test and Dunns, both with a significance level of 5% (p ≤ 0.05). On the planktonic cultures, CIM and CBM of extracts was determined only to F. nucleatum. The CBM of the extracts of A. satureioides, C. scolymus and P. americana was obtained on P. micra. Bactericidal activity was not obtained for P. endodontalis and P. gingivalis. About biofilms, all species exhibited significant reductions when exposed to the extracts in both times. It can be concluded that the extracts tested showed bacteriostatic effect on F. nucleatum. Bactericidal activity of extracts was observed on F. nucleatum and P. micra, except for H. virginiana. The extracts evaluated also presented antibiofilme effect on F. nucleatum, P. micra, P. endodontalis and P. gingivalis for 5 min and 24 h(AU)

Absceso cerebral en niños: experiencia en diez años en un hospital pediátrico de alta complejidad / Brain abscess in children: ten years of experience in a third level pediatric hospital

El absceso cerebral es una infección focal, que se presenta con una frecuencia de 0,3-1,3 casos cada 100000 personas/año.¹ Se describe la epidemiología, clínica y microbiología de 38 niños con diagnóstico de absceso cerebral internados entre el 1/4/2005 y el 31/12/2015 en el Hospital de Pediatría "Prof. Dr. Juan P. Garrahan" de la Ciudad de Buenos Aires. Veinticuatro pacientes fueron varones. La mediana de edad fue de 132 meses. Se detectaron factores predisponentes en 25 niños. La mediana de evolución entre el inicio de los síntomas y la consulta fue de 7 días. Hubo 27 casos con lesiones únicas. Se realizó un drenaje quirúrgico en 34 pacientes. Las bacterias más frecuentes fueron anaerobios, Streptococcus viridans y Staphylococcus aureus. La mediana de tratamiento antibiótico fue de 56 días y la mediana de internación fue de 43 días. La letalidad fue del 3%.

Brain abscess is a focal infection that occurs with a frequency of 0.3-1.3 cases per 100,000 people/year. We describe the epidemiology, clinical and microbiology characteristics of 38 children diagnosed with brain abscess hospitalized between 4/1/2005 and 12/31/2015 at Hospital de Pediatría "Prof. Dr. Juan P. Garrahan" in Buenos Aires City. Twenty-four patients were male. The median age was 132 months. Predisposing factors were detected in 25 children. The median evolution from onset of symptoms to the visit was 7 days. There were 27 cases with single lesions. Surgical drainage was performed in 34 patients. The most frequent bacteria were anaerobes, Streptococcus viridans and Staphylococcus aureus. The median of antibiotic treatment was 56 days and the median of hospitalization was 43 days. The lethality was 3%.

Presencia de bacilos negro pigmentantés en bolsas periodontales de pacientes diabéticos tipo 2 y su relación con el control glicémico / Presence of bacilli black pigments in periodontal pockets of patients with type 2 diabetes and its relationship with the glycemic level

Objetivos: El estudio tuvo como objetivo identificar la presencia de bacilos negro pigmentantés (BNP) en bolsas periodontales de pacientes diabéticos tipo 2 y determinar una relación con su control glicémico. Material y métodos: Se evaluó un número total de 46 pacientes que cumplían con los criterio de inclusión en el estudio, de los cuales quedaron 20 por haber concluido con todas las pruebas realizadas en el estudio, como el estudio microbiológico de sus muestras de placa subgingival y las pruebas de hemoglobina glicosilada que se realizó en forma periódica en los 3 meses que se evaluó a los pacientes. Resultados: De los 20 pacientes evaluados que completaron todas sus pruebas, 14 eran mujeres y 6 varones, con un rango de edad de 40 a 68 años, pudiéndose identificar estos bacilos negro pigmentantés (BNP) en el 20 % de casos con diagnóstico de periodontitis moderada a severa (periodontitis moderada: bolsa periodontal de 6 a 7 mm y Periodontitis severa: bolsa periodontal mayor de 7 mm), presentando estos pacientes un control glicémico de condición bueno a moderado (control bueno: = 6.9 % HbAc1y control moderado: 7 a 7.9 % HbAc1). La prueba estadística realizada para valorar la relación presencia BNP y el control glicémico fue de Chi cuadrado el cual no determino significancia de la relación (P > 5 %). Conclusiones: Se identificó la presencia de BNP en pacientes diabéticos tipo 2 pero más relacionado a pacientes con cuadros de periodontitis moderada a severa y con un control glicémico de bueno a moderado, no presentando significancia estadística.

Objectives: The study aimed to identify the presence of bacilli black pigments in periodontal pockets of type 2 diabetic patients and determine a relationship with glycemic control. Material and methods: A total of 46 patients who met the criteria for inclusion in the study, of which were 20 for having completed all tests in the study, such as microbiological analysis of samples of subgingival plaque and evaluated glycosylated hemoglobin tests held periodically in the 3 months patients were assessed. Results: Of the 20 patients evaluated, 14 were women and 6 men, with an age range of 40 to 68 years, being able to identify these black bacilli pigments (BNP) in 20% of cases diagnosed with moderate to severe periodontitis (periodontitis. moderate: periodontal pocket of 6 to 7 mm and severe periodontitis: greater periodontal pocket of 7 mm), presenting these patients glycemic control in good condition to moderate (good control: = 6,9 % HbAc1 and moderate control: 7 to 7,9 % HbAc1) . The statistical test performed to assess the presence relationship BNP and glycemic control was Chi square which does not determine significance of the relationship (P> 5%). Conclusions: It was possible to identify the presence of BNP in patients with type 2 diabetes but more related to patients with moderate to severe pictures periodontitis and with good glycemic control to moderate, showing no statistical significance.

Concentración de nitritos y nitratos en embutidos elaborados en Misiones y su relación con el crecimiento de Anaerobios reductores de sulfito / Concentration of nitrites and nitrates in sausages made in Misiones and its relationship with the growth of sulfite-reducing anaerobes

Los nitritos y nitratos son conservantes alimentarios utilizados para evitar la proliferación de micoorganismos anaerobios, incluido Clostridium botulinum, que produce la toxina causante del botulismo, enfermedad transmitida por alimentos con alta letalidad. Asimismo estos conservantes pueden tener efectos tóxicos agudos, causando en el consumidor metahemoglobinemia como en el caso de los nitratos, y efectos crónicos, mediante formación de nitrosaminas en el caso de los nitritos, consideradas potentes carcinógenos, por lo que deben usarse bajo límites establecidos en la legislación. El objetivo de esta investigación fue determinar la concentración de nitratos y nitritos en embutidos elaborados en la provincia de Misiones, Argentina, y su relación con el crecimiento de anaerobios reductores de sulfito. El estudio fue descriptivo de corte transversal. Tras determinación de tamaño muestral y selección aleatoria de participantes, previo consentimiento de los elaboradores, se tomaron muestras de embutidos elaborados en la provincia de Misiones con posterior determinación de nitratos y nitritos y recuento de anaerobios reductores de sulfito en el Laboratorio, mediante metodología oficial. De 58 muestras estudiadas, solo una (1,7%) excedió el límite establecido para la concentración de Nitritos y ninguna para las concentraciones de Nitratos. En cuanto a la relación entre el crecimiento de bacterias anaerobias reductoras de sulfito y la concentración de nitrito, se observó que hay una relación inversamente proporcional entre las mismas demostrado estadísticamente (χ2 = 31.48 y p < 0.05). Estos resultados contribuyeron al conocimiento sobre los parámetros antes descritos, al desarrollo de estrategias tanto de vigilancia como de asesoramiento hacia los elaboradores, con la finalidad de lograr un mejoramiento de las buenas prácticas de manufactura y de la elaboración de los embutidos en la provincia

Avaliação da percepção pessoal em relação à condição de halitose e confirmação clínica / Personal perception of assessment for the condition of halitosis and clinical confirmation

Introdução: A halitose é uma condição em que o hálito se apresenta de forma desagradável, acometendo milhares de pessoas e causando impacto negativo para as mesmas. Objetivo: A presente pesquisa em campo buscou avaliar a percepção pessoal em relação à condição de halitose por meio de aplicação de questionário específico e confirmação clínica. Material e método: A amostra foi composta por 59 indivíduos entre 20 e 60 anos de idade, usuários de Unidades Básicas de Saúde de Araraquara, que foram selecionados de forma aleatória. A metodologia empregada constituiu-se de: i) Assinatura do Termo de Consentimento Livre e Esclarecido; ii) Aplicação de questionário especialmente elaborado para esta pesquisa; iii) Confirmação clínica, com a aplicação do teste com halímetro (Tanita Breath Alert CHC-212S). Após o experimento, todos receberam folhetos informativos com dados gerais sobre halitose. Os resultados obtidos foram agrupados em tabelas de contingência, construídas pelo Microsoft Excel, sendo então analisados pelo teste do Qui-quadrado. Resultado e conclusão: Após análises, conclui-se que as pessoas perderam a capacidade de perceber o próprio hálito quando tinham halitose; não tiveram condições de se autoavaliarem de forma eficaz; os fatores de riscos que influenciaram na halitose foram: idade superior aos 40 anos, uso de alguns tipos de medicamentos, algumas patologias, frequência de visita ao Dentista e falhas em procedimentos de higiene bucal, tais como escovação dentária, uso do fio dental, limpeza da língua e limpeza de próteses. Os outros fatores pesquisados não se mostraram estatisticamente suficientes para estabelecerem relação de influência na halitose.

Introduction: Halitosis is a condition characterized by unpleasant breath,affecting thousands of people and causing a negative impact on them. Objective: The aim of this study was to evaluate self-perception of halitosis condition by applying a specific questionnaire and having clinical evaluation. Material and method: The sample was composed by 59 randomly selected voluntaries between the ages of 20 and 60 years old who were users of Basic Health Units of Araraquara. The methodologies applied were: 1. Signature of informed consent; 2. Application of standardized questionnaire specially designed for this research; 3. Clinical confirmation through halimeter (Tanita Breath Alert CHC-212S). After the experimental phase, all the volunteers received a document with general information about halitosis (causes and treatment). The results were grouped in Microsoft Excel tables and analyzed by Chi-square test. Result and conclusion: After the analyses, it was concluded that people had lost the ability to perceive their own breath when they had halitosis and they are unable to evaluate themselves effectively; the risky factors that influenced the halitosis were: people over the age of 40, the use some types of drugs, some diseases, frequency of visit to the dentist and poor oral hygiene including bad-technique procedures such as teeth brushing, flossing, tongue cleaning and dentures cleaning. Other factors studied were not statistically sufficient to influence halitosis.

Bacterias anaerobias: procesos que realizan y contribuyen a la sostenibilidad de la vida en el planeta / Anaerobic bacteria: processes they perform and their contribution to life sustainability on the planet

Objetivo. Se realiza una revisión sobre los aspectos degradativos de materia orgánica, obtención de energía y nutrientes de las bacterias anaerobias. La importancia de estos microorganismos es el papel que desempeñan en los procesos que contribuyen al mantenimiento de la vida misma. Dentro del metabolismo para la descomposición de macromoléculas, estos microorganismos realizan varios procesos: hidrólisis, acetogénesis y metanogénesis, entre otros, cobija reacciones que se realizan dependiendo de las características particulares de la bacteria y de las funciones que cumplen dentro del ciclo degradativo, para la obtención de nuevos productos dependiendo de las rutas bioquímicas o procesos fermentativos que allí se desarrollan.

Objective. A review of the degradation aspects of organic matter, obtaining energy and nutrients from anaerobic bacteria is performed. The importance of these organisms is the role they play in the processes contributing to the maintenance of life itself. Within the metabolism for the decomposition of macromolecules, these microorganisms perform several processes: hydrolysis acetogenesis and methanogenesis, among others, it covers reactions performed depending on the particular characteristics of the bacteria and of the roles in the degradative cycle for development of new products depending on the biochemical pathways or fermentation processes that take place there.

[Anaerobic bacteria 150 years after their discovery by Pasteur]. / Las bacterias anaerobias 150años después de su descubrimiento por Pasteur.

In 2011 we celebrated the 150th anniversary of the discovery of anaerobic bacteria by Louis Pasteur. The interest of the biomedical community on such bacteria is still maintained, and is particularly focused on Clostridium difficile. In the past few years important advances in taxonomy have been made due to the genetic, technological and computing developments. Thus, a significant number of new species related to human infections have been characterised, and some already known have been reclassified. At pathogenic level some specimens of anaerobic microflora, that had not been isolated from human infections, have been now isolated in some clinical conditions. There was emergence (or re-emergence) of some species and clinical conditions. Certain anaerobic bacteria have been associated with established infectious syndromes. The virulence of certain strains has increased, and some hypotheses on their participation in certain diseases have been given. In terms of diagnosis, the routine use of MALDI-TOF has led to a shortening of time and a cost reduction in the identification, with an improvement directly related to the improvement of data bases. The application of real-time PCR has been another major progress, and the sequencing of 16srRNA gene and others is currently a reality for several laboratories. Anaerobes have increased their resistance to antimicrobial agents, and the emergence of resistance to carbapenems and metronidazole, and multi-resistance is a current reality. In this situation, linezolid could be an effective alternative for Bacteroides. Fidaxomicin is the only anti-anaerobic agent introduced in the recent years, specifically for the diarrhoea caused by C.difficile. Moreover, some mathematical models have also been proposed in relation with this species.

Role of endotoxin in the etiology of periapical lesions: molecular mechanisms involved in endotoxin's recognition and cell activation / Papel da endotoxina na etiologia da lesão periapical: mecanismos moleculares envolvidos no seu reconhecimento e na ativação celular

Periapical lesions are diseases resulting from contamination and necrosis of dental pulp and the progression of this infection toward the periapical tissues. Technological advances in microbiological culture and identification have shown that anaerobic microorganisms, especially Gram-negative, are predominate in root canals of teeth with pulp necrosis and chronic periapical lesions. Gram-negative bacteria not only have different factors of virulence and generate products and sub-products that are toxic to apical and periapical tissues, but also contain endotoxin in the outer membrane of their cell wall. This information is important because endotoxin is released during multiplication or bacterial death, causing a series of biological effects that lead to an inflammatory reaction and resorption of mineralized tissues. Thus, the aim of this work is to carry out a review of the literature on the role of bacterial endotoxin in the etiology of periapical lesions, its mechanism of action, and to elucidate molecular mechanisms involved in endotoxin's recognition by the immune system and cell activation.

As lesões periapicais são patologias resultantes da contaminação e necrose da polpa dental e progressão da infecção em direção aos tecidos periapicais. Os avanços tecnológicos na cultura e identificação microbiana demonstraram que, em canais radiculares de dentes portadores de necrose pulpar e lesão periapical crônica, predominam os microrganismos anaeróbios, particularmente os Gram-negativos. As bactérias Gram-negativas, além de possuírem diferentes fatores de virulência e gerarem produtos e subprodutos tóxicos aos tecidos apicais e periapicais, contêm a endotoxina em sua parede celular. Esse conhecimento é importante, uma vez que a endotoxina é liberada durante a multiplicação ou morte bacteriana, exercendo uma série de efeitos biológicos relevantes, que conduzem a uma reação inflamatória e à reabsorção dos tecidos mineralizados. Sendo assim, o objetivo deste trabalho foi efetuar uma revisão da literatura relativa ao papel da endotoxina bacteriana na etiologia das lesões periapicais, seu mecanismo de ação, bem como elucidar mecanismos moleculares envolvidos no seu reconhecimento pelo sistema imunológico e na ativação celular.

[Utility of MALDI-TOF MS for the identification of anaerobic bacteria]. / Utilidad de la espectrometría de masas MALDI-TOF en la identificación de bacterias anaerobias.

The analysis by MALDI-TOF MS (Matrix-assited laser desorption/ionization time-of-flight mass spectrometry) has become a reference method for the identification of microorganisms in Clinical Microbiology. However, data on some groups of microorganisms are still controversial. The aim of this study is to determine the utility of MALDI-TOF MS for the identification of clinical isolates of anaerobic bacteria. One-hundred and six anaerobic bacteria isolates were analyzed by MALDI-TOF MS and by conventional biochemical tests. In those cases where identification by conventional methodology was not applicable or in the face of discordance between sequencing methodologies, 16 S rRNA gene sequence analysis was performed. The conventional method and MALDI-TOF MS agreed at genus and species level by 95.3 %. Concordance in gram-negative bacilli was 91.4% and 100% among gram-positive bacilli; there was also concordance both in the 8 isolates studied in gram-positive cocci and in the single gram-negative cocci included. The data obtained in this study demonstrate that MALDI-TOF MS offers the possibility of adequate identification of anaerobic bacteria.