Antimicrobial action of psidium guajava l. extract against enterococcus faecalis and candida albicans strains: a pilot study / Ação antimicrobiana do extrato de psidium guajava l. contra as cepas de enterococcus faecalis e candida albicans: um estudo piloto / Acción antimicrobiana de la extracción de psidium guajava l. contra los cepes de los enterococcus faecalis y los candida albicans: un estudio piloto

Endodontic treatment consists of decontamination of the root canal using irrigants like Sodium Hypochlorite (NaOCl). However, some microorganisms resist conventional therapies, causing relapse and secondary infections. Substances with microbicidal power and potential to become new irrigants need to be studied. Thus, the objective of the present study was to evaluate the antimicrobial potential of Psidium guajava L. (guava tree) extract against clinical and standard (ATCC) strains of Enterococcus faecalis and Candida albicans. For this purpose, the extract was produced from the raw material (leaves and shoots) diluted in the hydroethanolic vehicle (EtOH: H2O / 50:50) for subsequent microbiological analysis. The Minimum Inhibitory Concentration (MIC) and Minimum Microbicidal Concentration (MMC) of the plant extract were tested, according to the Clinical and laboratory standards institute (CLSI) guidelines, on four strains: E. faecalis (ATCC and clinic) and C. albicans (ATCC and clinic). The extract of P. guajava did not produce antifungal action on C. albicans, however, it showed microbicidal potential against strains of E. faecalis, showing MIC of 0.20%. This concentration was lower than the MIC of NaOCl which was 0.31%, a solution that is commonly used in dental clinics. In conclusion, the hydroethanolic extract of P. guajava presents bactericidal action against E. faecalis, being a natural product with potential for future studies regarding the development of new endodontic irrigants.

O tratamento endodôntico consiste na descontaminação do canal radicular com emprego de soluções irrigadoras, como por exemplo, o Hipoclorito de Sódio (NaOCl). Contudo, alguns microrganismos resistem as terapias convencionais, causando recidiva e infecções secundárias. Substâncias com poder microbicida e potencial para se tornarem novas soluções irrigadoras precisam ser estudadas. Assim, o objetivo do presente estudo foi avaliar o potencial antimicrobiano do extrato de Psidium guajava L. (goiabeira) contra cepas clínicas e padrão (ATCC) de Enterococcus faecalis e Candida albicans. Para tanto, o extrato foi produzido a partir da matéria bruta (folhas e brotos) diluído no veículo hidroetanólico (EtOH: H2O / 50:50) para posterior análise microbiológica. Foram testadas a Concentração Inibitória Mínima (CIM) e Concentração Microbicida Miníma (CMM) do extrato vegetal, de acordo com as normas da Clinical and laboratory standards institute (CLSI), sobre quatro cepas: E. faecalis (ATCC e clínica) e C. albicans (ATCC e clínica). O extrato de P. guajava não produziu ação antifúngica sobre C. albicans, contudo, apresentou potencial microbicida contra cepas de E. faecalis, exibindo CIM de 0,20%. Esta concentração foi menor que a CIM do NaOCl que foi de 0,31%, uma solução que é comumente utilizada nas clínicas odontológicas. Em conclusão, o extrato hidroetanólico de P. guajava apresenta ação bactericida contra E. faecalis, sendo um produto natural com potencial para futuros estudos quanto ao desenvolvimento de novos irrigantes endodônticos.

El tratamiento endotónico consiste en la descontaminación del canal raíz con el uso de soluciones de irrigación, como el Hipoclorito de Sodio (NaOCl). Sin embargo, algunos microorganismos se resisten a las terapias convencionales, causando recaídas e infecciones secundarias. Es necesario estudiar las sustancias con poder de microbicida y el potencial para convertirse en nuevas soluciones de riego. Por lo tanto, el objetivo de este estudio fue evaluar el potencial antimicrobiano del extracto de Psidium guajava L. (guava) frente a cepas clínicas y estándar (ATCC) de Enterococcus faecalis y Candida albicans. Con este fin, el extracto se produjo a partir de la materia prima (hojas y brotes) diluida en el vehículo hidroetanólico (EtOH: H2O / 50:50) para análisis microbiológicos posteriores. La Concentración Mínima Inhibidora (CMI) y la Concentración de Microbicidas Minima (CCM) del extracto vegetal se probaron de acuerdo con el Clinical and laboratory standards institute (CLSI) en cuatro cepas: E. faecalis (ATCC y clínico) y C. albicans (ATCC y clínico). El extracto de P. guajava no produjo acción antifúngica sobre C. albicans, sin embargo mostró potencial microbicida frente a cepas de E. faecalis, presentando CIM 0,20%. Esta concentración fue menor que la CMI de NaOCl, que fue del 0,31%, una solución comúnmente utilizada en clínicas dentales. En conclusión, el extracto hidroetanólico de P. guajava muestra una acción bactericida frente a E. faecalis, siendo un producto natural con potencial para futuros estudios sobre el desarrollo de nuevos irrigantes endotónicos.

Molecular and biotechnological characteristics of proteolytic activity from Streptococcus thermophilus as a proteolytic lactic acid bacteria to enhance protein-derived bioactive peptides.

The demand for healthy food items with a high nutrient value of bioavailability and bioaccessibility has created a need for continuous development of technology and food ingredients like bioactive peptides. This study aimed to investigate seven proteolytic lactic acid bacteria (PLABs) isolated from the plaa-som (fermented fish) sample originated from silver BARB species for production of proteolytic enzymes. Proteolytic enzymes produced by (PLABs) were used further to create potent bioactive peptides by hydrolyzing proteins throughout PLAB-probiotics enhancer. Protein derived-bioactive peptides was tested the proteolytic activity on different protein sources and examined bioactivities including antioxidative and antimicrobial effect for further use in functional foods. Results of screened-PLAB strains showed high proteolytic activity namely Streptococcus thermophilus strains (KKUPA22 and KKUPK13). These strains have proteolytic system consisting of extracellular and cell-bound enzymes that used for degrading protein in fish flesh protein (FFP) and skim milk (SKM) broth media. Proteolytic activity of tested bacterial enzymes was estimated after incubation at 45, 37, and 50 °C. Furthermore, FFP hydrolysates were formed with various peptides and has small molecular weights (checked by SDS-PAGE) in the range of10.5 to 22 kDa), exhibiting strong activity. Data revealed that S. thermophilus strains (KKUPA22 and KKUPK13) had high antioxidant activity in term of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS) radical-scavenging inhibition, and ferric reducing antioxidant power (FRAP) reducing power capacity. Both strains (KKUPA22 and KKUPK13) of S. thermophilus have higher antimicrobial activity against Gram-negative bacteria than against Gram-positive bacteria. We have confirmed presence of proteolytic (prt) gene regions in S. thermophilus strains using specific primers via PCR amplification. Results showed highest homology (100%) with the prtS gene of S. thermophillus located on the cell envelope proteolytic enzymes (CEPEs) such as serine proteinase. Therefore, it concluded that the proteolytic system of tested PLAB strains able to generate bioactive peptides-derived proteins having active biological property, good mechanism of degradability, and bioaccessibility for further use in catalyzing protein of functional foods.

Augmented reality navigation with ultrasound-assisted point cloud registration for percutaneous ablation of liver tumors.

PURPOSE: We present a novel augmented reality (AR) surgical navigation method with ultrasound-assisted point cloud registration for percutaneous ablation of liver tumors. A preliminary study is carried out to verify its feasibility. METHODS: Two three-dimensional (3D) point clouds of the liver surface are derived from the preoperative images and intraoperative tracked US images, respectively. To compensate for the soft tissue deformation, the point cloud registration between the preoperative images and the liver is performed using the non-rigid iterative closest point (ICP) algorithm. A 3D AR device based on integral videography technology is designed to accurately display naked-eye 3D images for surgical navigation. Based on the above registration, naked-eye 3D images of the liver surface, planning path, entry points, and tumor can be overlaid in situ through our 3D AR device. Finally, the AR-guided targeting accuracy is evaluated through entry point positioning. RESULTS: Experiments on both the liver phantom and in vitro pork liver were conducted. Several entry points on the liver surface were used to evaluate the targeting accuracy. The preliminary validation on the liver phantom showed average entry-point errors (EPEs) of 2.34 ± 0.45 mm, 2.25 ± 0.72 mm, 2.71 ± 0.82 mm, and 2.50 ± 1.11 mm at distinct US point cloud coverage rates of 100%, 75%, 50%, and 25%, respectively. The average EPEs of the deformed pork liver were 4.49 ± 1.88 mm and 5.02 ± 2.03 mm at the coverage rates of 100% and 75%, and the average covered-entry-point errors (CEPEs) were 4.96 ± 2.05 mm and 2.97 ± 1.37 mm at 50% and 25%, respectively. CONCLUSION: Experimental outcomes demonstrate that the proposed AR navigation method based on US-assisted point cloud registration has achieved an acceptable targeting accuracy on the liver surface even in the case of liver deformation.

Online Cure Monitoring and Modelling of Cyanate Ester-Based Composites for High Temperature Applications.

A cure kinetics investigation of a high temperature-resistant phenol novolac cyanate ester toughened with polyether sulfone (CE-PES blend) was undertaken using non-isothermal differential scanning calorimetry. Thin ply carbon fiber prepreg, based on the CE-PES formulation, was fabricated, and plates for further in-situ cure monitoring were manufactured using automated fiber placement. Online monitoring of the curing behavior utilizing Optimold sensors and Online Resin State software from Synthesites was carried out. The estimation of the glass transition temperature and degree of cure allowed us to compare real time data with the calculated parameters of the CE-PES formulation. Alongside a good agreement between the observed online data and predicted model, the excellent performance of the developed sensors at temperatures above 260 °C was also demonstrated.

Quantificação e identificação de Staphylococcus coagulase positiva e negativa em leites cru e pasteurizado comercializados na cidade de São Luís ­ MA / Quantification and identification of positive and negative Staphylococcus coagulase in raw and pasteurized milks marketed in the city of São Luís - MA

A qualidade microbiológica deleites cru e pasteurizado produzidos e comercializados em São Luís ­ MA foi avaliada por meio da quantificação e identificação de cepas de micro-organismos do gênero Staphylococcus. Verificou-se um alto nível de contaminação por Staphylococcus sp. em 60% das amostras de leite cru analisadas, com as contagens variando entre 1,7x105 e 1,1x107 UFC/mL, sendo comprovada a presença de Staphylococcus coagulase positiva nessas amostras. No leite pasteurizado foram obtidas baixas contagens desse micro-organismo, com valores variando entre 2,7x102 e 4,2x102 UFC/mL, não sendo detectados Staphylococcus coagulase positiva, apenas coagulase negativa em 23% (n=3) das amostras analisadas. As colônias características de Staphylococcus isoladas foram identificadas por meio de testes bioquímicos convencionais. Das 50 cepas isoladas, 38% foram positivas para o teste de coagulase, sendo identificadas as espécies S. aureus, S. hyicus, S. intermedius e S. delphini. Entre as cepas de Staphylococcus coagulase negativa, verificou-se a predominância das espécies S. gallinarum, S. cloosii, e S. xylosus. A presença de tais micro-organismos no leite é preocupante do ponto de vista da segurança dos alimentos, tendo em vista que espécies como S. aureus, S. hyicus e S. intermedius são potenciais produtoras de enterotoxinas termoestáveis.

The microbiological quality of raw and pasteurized milk produced and sold in São Luís ­ MA was evaluated through quantification and identification of Staphylococcus cepes. A high level contamination by Staphylococcus sp. was verified in 60% of samples of raw milk, counting varying between 1.7 × 105 e 1.1 × 107 CFU/mL, which proves the presence of positive coagulasis Staphylococcus in these samples. In the pasteurized milk low counting of this microorganism was obtained, values varying between 2.7 × 102 e 4.2 × 102 CFU/mL, in where it was not found positive coagulase Staphylococcus, but only negative ones in 23% (n=3) of the analyzed samples. The typical colonies of isolated Staphylococcus were identified through conventional biochemical tests. From 50 isolated cepes, 38% were positive for coagulase test, in which S. aureus, S. hyicus, S. intermedius e S. delphini species were identified. Among the cepes of negative coagulase Staphylococcus6, S. gallinarum, S. cloosii, e S. xylosus were predominant. The presence of these microorganisms in milk is a motive to be worried about concerning food safety, for species like S. aureus, S. hyicus e S. intermedius are potential producers of thermostable enterotoxins.

[The composition of volatile components of cepe (Boletus edulis) and oyster mushrooms (Pleurotus ostreatus)].

The composition of aroma compounds in cooked and canned cepe (Boletus edulis) and in cooked oyster mushrooms (Pleurotus ostreatus) is studied using capillary gas chromatography and chromatography-mass spectrometry. It is found that unsaturated alcohols and ketones containing eight atoms of carbon determine the aroma of raw mushrooms and take part in the formation of the aroma of cooked mushrooms as well. The content of these compounds was the highest in canned cepes. In oyster mushrooms, the concentration of these alcohols and ketones was lower in comparison with cepes. The content of aliphatic and aromatic aldehydes was much higher in oyster mushrooms. Volatile aliphatic and heterocyclic Maillard reaction products and isomeric octenols and octenones formed the aroma of cooked and canned mushrooms.

Infecciones invasivas por Staphylococccus aureus meticilino resistente adquirido en la comunidad. Presentación clínica y evolutiva observada en dos centros universitarios. Uruguay 2003-2007 / Invasive community-acquired methicillin-resistant Staphylococcus aureus infections. Clinical and evolutionary presentation observed in two university centers. Uruguay 2003-2007

Introducción: en Uruguay, desde el año 2001 Staphylococcus aureus meticilino resistente no multirresistente (SAMR-AC) se estableció como un patógeno emergente en las infecciones comunitarias del niño. Entre el año 2003 y 2004, la rápida diseminación de las infecciones por este germen ocasionó la muerte de varias personas generando alarma sanitaria. Objetivo: describir las características clínicas y evolutivas de los niños hospitalizados con infecciones invasivas por SAMR-AC en dos centros universitarios en el período 2003-2007. Material y método: se incluyeron los niños con infecciones invasivas por SAMR-AC hospitalizados entre el 1º de enero de 2003 y el 31 de diciembre de 2007 en el Hospital Pediátrico del Centro Hospitalario Pereira Rossell (HP-CHPR) y en el Hospital Escuela del Litoral de Paysandú (HELP). Se definió infección invasiva cuando el germen fue aislado de sitios habitualmente estériles. Se analizó edad, sexo, topografía, complicaciones y mortalidad. Resultados: fueron hospitalizados 78 niños, edad promedio 5,4 años. Las presentaciones clínicas más frecuentes fueron empiema (n=28), infección osteoarticular (n=27) y bacteriemia secundaria a infección de piel y partes blandas (n=9). En los niños con empiema, presentaron sepsis o shock séptico, o ambos, cinco; requirieron drenaje de tórax 27 niños, de los cuales 11 presentaron abscesos de pared, seis osteomielitis costal y uno requirió neumonectomía parcial. En el grupo con infecciones osteoarticulares, todos requirieron drenaje; presentaron pandiafisitis nueve; sepsis o shock séptico, o ambos, siete; trombosis venosa profunda cinco, y evolucionaron a la cronicidad cinco. Fallecieron cinco niños (6,4%); tres con empiema y dos con infecciones osteoarticulares. Conclusiones: una proporción importante de los niños con infecciones invasivas por SAMR-AC presentaron secuelas; la mortalidad en esta serie fue 6,4%. El conocimiento de sus formas de presentación clínica y posible evolución, resulta indispensable para el diagnóstico precoz y tratamiento empírico adecuado. La caracterización genética de las cepas podría contribuir a mejorar el control y el tratamiento.

Summary Introduction: in Uruguay, since 2001, community acquired non-multiresistant methicillin-resistant staphylococcus aureus was defined as an emerging pathogen in child community acquired diseases. Between 2003 and 2004, the fast dissemination of infections caused by this germ resulted in several people dying and the uprising of a health alarm. Objective: to describe the main clinical features and evolution in children hospitalized due to community acquired methicillin resistant staphylococcus aureus in two university medical centers between 2003 and 2007. Method: the study was conducted in children with invasive community acquired methicillin resistant staphylococcus aureus infections, who were hospitalized in the Pediatric Hospital of the Pereira Rossell Health Care Center and in the Litoral de Paysandú Teaching Hospital between January 1, 2003 and December 31, 2007. We defined an infection as invasive when the germ was isolated in usually sterile sites. We analyzed age, sex, topography, complications and mortality. Results: 78 children were hospitalized; average age was 5.4 years old. The most common clinical presentations were empyema (n=28), osteoarticular infection (n=27), bacteremia secondary to skin and soft tissue infections (n=9). Among children with empyemia, 5 evidenced sepsis or septic shock, or both; 27 required thorax drain, 11 of which evidenced abcess in the chest cavity, 6 of which evidenced costal osteomyelitis, one of which required partial neumonectomy. Among the osteoarticular infection group, they all required drainage; 9 evidenced pandiaphystis, sepsis or septic sic, or both; 5 evidenced deep vein thrombosis, and 5 evolved to chronicity. Five children (6.4%) died; three of them with empyemia and two with osteoarticular infections Conclusions: a large proportion of children with invasive community acquired methicillin resistant staphylococcus aureus infections presented sequelae; mortality in this series was 6.4%. Learning about the different forms of clinical presentation and possible evolution is essential for early diagnosis and the appropriate empirical treatment. Genetic characterization of strains could contribute to improving control and treatment.

Résumé Introduction: en Uruguay,depuis 2001, Staphylococcus aureus méticilline résistant non multi-résistant (SAMR-AC) s’est établi comme pathogène émergeant aux infections communautaires chez l’enfant. Entre 2003-2004, la rapide dissémination des infections par ce germe a provoqué la mort de plusieurs personnes, ce qui a motivé une alarme sanitaire. Objectif: décrire les caractéristiques cliniques et évolutives des enfants hospitalisés avec infections invasives par SAMR-AC dans deux centres universitaires pendant la période 2003-2007. Matériel et méthode: on inclut les enfants à infections invasives par SAMR-AC hospitalisés entre le 1er janvier 2003 et le 31 décembre 2007 à l’Hôpital Pédiatrique du Centre Hospitalier Pereira Rossell (HP-CHPR) et à l’Hôpital École du Litoral de Paysandú (HELP). On détermine infection invasive lorsque le germe est isolé dans des sites habituellement stériles. On tient compte de l’âge, du sexe, de la topographie, des complications et de la mortalité. Résultats: 78 enfants sont hospitalisés, dont une moyenne d’âge 5,4 ans. Les manifestations cliniques les plus fréquentes ont été empyème (n=28), de peau et de parties molles (n=9). Chez les enfants à empyème, cinq ont présenté sepsis ou shock septique, ou les deux; 27 enfants ont requis drainage de thorax dont 11 abcès de paroi, six, ostéomyélite costale et un a subi une pneumonectomie partielle. Dans le groupe d’infections ostéo-articulaires, tous ont requis drainage; neuf ont présenté pandiaphysite; sept, sepsis ou shock septique, ou tous les deux; cinq, thrombose veineuse profonde et cinq ont évolué vers la chronicité. Cinq enfants ont décédé (6,4%); trois par empyème et deux à cause des infections ostéo-articulaires. Conclusions: un taux important des enfants avec infections invasives par SAMR-AC ont eu des séquelles; la mortalité fut de 6,4%. La connaissance de leurs manifestations cliniques et de leur possible évolution, s’avère indispensable pour le diagnostic précoce et un traitement empirique ajusté. La caractérisation génétique des cèpes pourrait contribuer à améliorer le contrôle et le traitement.

Resumo Introdução: no Uruguai, desde 2001, o Staphylococcus aureus resistente a meticilina não multi-resistente (SAMR-AC) está instalado como um agente patológico nas infecções comunitárias infantis. Entre 2003 e 2004, a rápida disseminação das infecções causadas por esse microorganismo causou a morte de várias pessoas gerando um estado de alerta sanitário. Objetivo: descrever as características clínicas e evolutivas das crianças internadas com infecções invasoras por SAMR-AC em dois centros universitários de atenção de saúde no período 2003-2007. Material e método: foram incluídas todas as crianças com infecções invasoras por SAMR-AC internados no período entre 1º de janeiro de 2003 e 31 de dezembro de 2007, no Hospital Pediátrico do Centro Hospitalar Pereira Rossell (HP-CHPR) e no Hospital Escola do Litoral de Paysandú (HELP). Infecção invasora foi definida como aquela em que o gérmen foi isolado de localizações normalmente estéreis. As variáveis analisadas foram: idade, sexo, topografia, complicações e mortalidade. Resultados: foram internadas 78 crianças, com idade média de 5,4 anos. As apresentações clínicas mais freqüentes foram: empiema (n=28), infecção osteoarticular (n=27) e bacteriemia secundária a infecção de pele e de partes moles (n=9). Entre as crianças que apresentaram empiema, cinco apresentaram choque séptico, sepse ou ambos; foi necessário realizar drenagem de tórax em 27, onze dos quais apresentavam abscessos de parede, seis osteomilite costal e em um caso foi realizada uma pneumonectomía parcial. No grupo com infecções osteoarticulares, foi necessário fazer drenagem em todos os casos; nove apresentaram pandiafisite, sepse ou choque séptico ou ambos; cinco trombose venosa profunda e cinco evoluíram a caso crônico. Cinco crianças faleceram (6,4%); três com empiema e duas com infecções osteoarticulares. Conclusões: uma proporção importante das crianças com infecções invasoras por SAMR-AC apresentou seqüelas; a mortalidade neste grupo de pacientes foi de 6,4%. É indispensável conhecer as diferentes formas de apresentação destas infecções e suas possíveis formas de evolução para a realização de um diagnóstico precoce e tratamento empírico adequado. A caracterização genética das cepas poderia ajudar a melhorar o controle e o tratamento.

Co-infecções Aids e Hepatite B e C / Co-infections Aids and Hepatitis B and C

Esta edição , que anuncia a Mesa Redonda sobre Co-infecçoes AIDS e Hepatite B e C, não só nos convida a fazer uma série de reflexões, como acabou transformando-se em uma verdadeira aula sobre a situação atual em nosso País, com propostas para a ação individual e coletiva do Setor Saúde como um todo. O local nao poderia ser mais feliz, o recem-inaugurado Centro de Pesquisa em Saúde, o CEPES, da Faculdade de Medicina do ABC. Os Drs. David Uip, Hoel Sette Jr., Roberto Focaccia e Maria Cassia Mendes Correa, além do altissimo nível dos debates, mantiveram durante toda a reunião uma atitude ousada, provocativa, questionando os seus próprios conhecimentos cientifícos com a sua vivência, do seu dia a dia na linha de frente da luta contra a AIDS e a hepatite. Podemos afirmar que ao término da reunião, todos os presentes sentiam que não se encerrava um debate, mas sim que algo novo começava a ser criado. Os debatedores não se limitaram a fazer as suas conclusões finais, mas plantaram idéias fundamentais que, sem duvida, vão nortear as açoes futuras que se fazem necessarias para colocar o nosso País, mais uma vez, em posição de destaque no campo da Medicina

Caracterización genotípica de 80 cepas del género Mycobacterium en Uruguay / Genotype characterization of 80 strains of Mycobacterium in Uruguay

Los métodos tradicionales de identificación fenotípica del género Mycobacterium son lentos y poco sensibles, requiriéndose cuatro a seis semanas para lograr un diagnóstico apropiado a partir de un cultivo positivo. Los procedimientos moleculares han permitido acortar este período, obteniéndose resultados entre las 36 a 72 horas. En nuestro país la incidencia de M. tuberculosis es baja y no existen datos acerca de con qué frecuencia los casos diagnosticados como tuberculosis pulmonar son en realidad causados por Mycobacterium no tuberculosis (MNT), normalmente saprofitas. Desde el punto de vista terapéutico, el diagnóstico etiológico a través de la identificación precisa de la especie de Mycobacterium infectante resulta un aporte significativo, dado que el tratamiento y el manejo de sus contactos son diferentes según sea la especie involucrada. Por estas razones se introdujo en nuestro laboratorio el diagnóstico de Mycobacterium a través de su identificación genotípica. Para ello se eligieron dos marcadores moleculares de ADN: la secuencia de inserción IS6110, característica de los genomas del complejo M. tuberculosis, y la secuencia del gen ribosomal 16s (ADNr 16s) para estudiar la identidad específica dentro del género Mycobacterium. Una vez puestas a punto las técnicas moleculares seleccionadas, se procedió al estudio retrospectivo de una colección de 80 aislamientos, identificados como Mycobacterium por métodos fenotípicos. La mayoría de los aislamientos (75/80) resultaron cepas del complejo M. tuberculosis. Los restantes cinco fueron identificados como cepas MNT, tres de ellas causantes de infecciones pulmonares.

Summary Traditional methods to determine phenotype identification for mycobacterium are longer as compared with cellular procedures (4 to 6 weeks and 36 to 72 hours respectively). In Uruguay, the incidence of tuberculosis Mycobacterium is low and data on pulmonary tuberculosis cases but caused by non-tuberculosis Mycobacterium (MNT) -normally saprophytes- is lacking. From a therapeutic point of view, diagnosis based on an accurate identification of Mycobacterium infectant may be significant since treatment and management differ according to the strain found. Two DNA molecular markers were chosen in our laboratory to diagnose Mycobacterium through genotype identification: IS6110 insertion element and ribosomal DNA sequences 16s (DNAr 16s) to determine specific identity within Mycobacterium. Once selected molecular techniques were updated, we undertook a retrospective study of 80 isolates identified as Mycobacterium by phenotype methods. Most of the isolates (75/80) were tuberculosis Mycobacterium strains. The remained five were identified as MNT strains, of which three caused pulmonary infections.

Résumé Les méthodes traditionnelles d'identification phénotypi-que du genre Mycobacterium sont lentes et peu sensibles, quatre à six semaines étant nécessaires pour avoir un diagnostic approprié à partir d'une culture positive. Les procédés moléculaires ont permis de raccourcir cette période: on obtient des résultants au bout de 36-72 heures. Dans notre pays, l'incidence de M. tuberculosis est basse et il n'y a pas de registres pour savoir la fréquence avec laquelle les cas diagnostiqués comme tuberculose pulmonaire sont en réalité causés par Mycobacterium non tuberculose (MNT), normalement saprophytes. Du point de vue thérapeutique, le diagnostic étiolo-gique à travers l'identification précise de l'espèce de mycobacterium infectante est un apport important, étant donné que le traitement varie selon l'espèce en question. Voilà pourquoi on a introduit dans notre laboratoire le diagnostic de Mycobacterium à travers son identification génotypique. Pour ce faire, on a choisi deux marqueurs moléculaires d'ADN: la séquence d'insertion IS6110, caractéristique des génomes du complexe M.tuberculose, et la séquence du gène ribosome 16s (ADN 16s) pour étudier l'identité spécifique dans le genre Mycobacterium. Une fois mises à point les techniques moléculaires sélectionnées, on fait une étude rétrospective d'une collec-tion de 80 isolements, identifiés comme Mycobacterium par des méthodes phénotypiques. La plupart des isolements (75/80) étaient des cèpes du complexe M. tuberculose. Les autres cinq ont été identifiés comme des cèpes MNT, dont trois étant la cause d'infections pulmonaires.