MALDI-TOF as a tool for microbiological monitoring in areas considered aseptic.

To maintain asepsis in production environments, contamination must be constantly controlled. To this end, microbiological monitoring is constantly used with the objective of evaluating the incidence of microorganisms prevalent in the sampling of air, surface, and people, in the area of an environment considered aseptic, isolated, and identified using the rapid and automated phenotypic microbiological methodology, highlighting the MALDI-TOF mass spectrometry analysis technique (MS), being identified at the level of genus and/or species. For that purpose, microbiological control of environmental monitoring of environments considered aseptic in a pharmaceutical industry was conducted for 12 months. The isolated microorganisms were identified using the mass spectrometry identification method (MALDI-TOF). In area classification A, the most prevalent microorganisms were bacteria in the sampling person. The microbial population was composed of bacteria of the genus Micrococcus sp. and Staphylococcus sp. Based on the results, it is possible to observe that in an environment where the process requires human operations, possible microbial contamination is inevitable and requires the identification of microorganisms at least at the level of species and/or genus. The microorganisms identified and found in the sampling of the aseptic environment must be evaluated with frequency to ensure that the productive environment guarantees the quality of the product produced.

Determinación de la eficacia de los desinfectantes empleados en las áreas asépticas de un centro productor de biofarmacéuticos / Determination of the efficacy of the disinfectants employed in the aseptic areas at a biopharmaceuticals production center

En la industria biofarmacéutica, la desinfección se ha hecho parte fundamental de las actividades para controlar el nivel de microorganismos contaminantes que puedan llegar a comprometer la calidad del producto final. El objetivo de este trabajo fue evaluar la eficacia de los desinfectantes AniosSpecial DJP SF, Aniosurf Premium, Bacteranios SF, Aniospray 29, Aseptanios AD y Surfanios utilizados en la limpieza y desinfección del área aséptica de la Planta de Productos Parenterales 2 del Centro Nacional de Biopreparados (Mayabeque, Cuba). Se comprobó la eficacia con pruebas de desafío en superficies y en condiciones prácticas de uso. Se comparó estadísticamente el número de microorganismos aislados y la microbiota presente en el área antes y después del uso de los desinfectantes. Todos los desinfectantes cumplieron con los requisitos establecidos. Por lo tanto, se demostró su eficacia para realizar la limpieza de las áreas asépticas(AU)

In the biopharmaceutical industry, disinfection has become a fundamental part of the activities to control the level of contaminants microorganisms that could compromise the quality of the final product. The objective of this work was to evaluate the efficacy of the disinfectants Anios Special DJP SF, Aniosurf Premium, Bacteranios SF, Aniospray 29, Aseptanios AD and Surfanios for the cleaning and disinfection of the aseptic area at the Parenterals Products Plant 2 (PPP2) at the National Center for Bioproducts (Mayabeque, Cuba). The efficacy was checked with challenge tests in surfaces, in practical usage conditions. The number of isolated microorganisms and the microbiota present before and after use was compared statistically. All the disinfectants complied with the established requirements. Therefore, the efficacy of the disinfectants was demonstrated to perform the cleaning and disinfection of the aseptic areas(AU)

Calidad microbiológica del aire de una unidad de preparados farmacéuticos estériles / Bacteriological quality of air in a ward for sterile pharmaceutical preparations

Introducción: La elaboración de preparados farmacéuticos estériles requiere áreas limpias que deben cumplir estándares internacionales para minimizar la contaminación microbiana. Objetivo: Evaluar la calidad bacteriológica del aire de la Unidad de Preparados Farmacéuticos Estériles del Servicio de Farmacia del Hospital Clínico de la Universidad de Chile y establecer niveles de alerta y acción. Material y Métodos: Se monitorearon ocho puntos representativos de la unidad, diariamente entre enero y febrero de 2005 y bisemanal-mente de junio a febrero de 2006. Se estudiaron 839 muestras de aire, recolectadas mediante el método de impacto en placa (equipo MAS-100). Resultados: De las muestras estudiadas, 474 (56,5 por ciento) fueron positivas; de éstas, sólo 17 (3,5 por ciento) estuvieron fuera del rango permitido, porcentaje que representa el 2 por ciento del total. Las muestras de los sitios 1 y 2 (flujo laminar grande y pequeño), que corresponden al área de preparación de preparados estériles fueron negativas. Los sitios 3 (mesón) y 4 (transfer) presentaron ocasionalmente valores superiores a los límites. Los microorganismos más frecuentes fueron Staphylococcus coagulasa negativa, Micrococcus spp y Corynebacterium spp, agentes de la microbiota de la piel y, menor porcentaje, Bacillus spp, agente de la microbiota ambiental. Conclusiones: Desde el punto de vista microbiológico, la calidad del aire de la zona de preparaciones estériles descrita presenta niveles ajustados a estándares internacionales. El establecer niveles de alerta y acción institucionales y la identificación de los microorganismos obtenidos en las áreas más sensibles de la unidad permite cuantificar la carga microbiana y conocer sus componentes para determinar las intervenciones a realizar cuando ellas estén indicadas.

Background: An extremely clean area is required for preparation of sterile pharmaceutical compounds, in compliance with international standards, to minimize the probability of microbial contamination. Aim: To evaluate the bacteriological quality of the air in the Sterile Pharmaceutical Preparation Unit of the University of Chile's Clinical Hospital and to set up alerts and action levels of bacterial growth. Methods: We studied eight representative sites of our Unit on a daily basis from January to February 2005 and twice a week from June 2005 to February 2006. We collected 839 samples of air by impact in the Petri dish. Results: 474 (56.5 percent) samples were positive; 17 (3.5 percent) of them had an inappropriate bacterial growth (2 percent of total samples). The samples from sites 1 and 2 (big and small biosafety cabinets) were negative. The countertop and transfer area occasionally exceeded the bacterial growth limits. The most frequently isolated bacteria were coagulase-negative staphylococci, Micrococcus spp and Corynebacterium spp, from skin microbiota, and Bacillus spp, an environmental bacteria. Conclusions: From a microbiological perspective, the air quality in our sterile preparation unit complied with international standards. Setting institutional alerts and action levels and appropriately identifying bacteria in sensitive areas permits quantification of the microbial load and application of preventive measures.

Coleta de amostras e métodos analíticos para determinaçãode chumbo / Collection of samples and analytical methods for lead determination

Chumbo é, frequentemente, usado como um indicador de poluição. Uma vez liberado no meio ambiente, este poluente pode seguir diferentes rotas, podendo ser convertido em espécies mais solúveis, tornado-se biodisponível. Neste artigo é apresentado de forma sucinta chumbo como poluente urbano, procedimentos de amostragem (coleta de amostras), transporte, armazenamento e preservação de amostras de interesse ambiental (solos e águas) e amostras clínicas (sangue e urina). A importância e classificação das Salas Limpas, assim como as principais técnicas analíticas para a determinação de chumbo - AAS, ICP OES e ICP-MS - também são abordadas.

The metal lead has frequently used as a sensitive pollution indicator. Once released in the environment, this pollutant can follow different ways, being converted in highly soluble species, became bioavailable. In this paper is presented in a succinct form, lead as urban pollutant, sampling procedures (collection of samples), transport, storage and preservation of environmental (soils and waters) and clinical (bloodand urine) samples. The importance and classification of clean rooms, as well as the main analytical techniques for lead determination - AAS, ICP-OES and ICP-MS - are broached too.

Microbiological monitoring of clean rooms in development of vaccines

The aim of the present work was to evaluate an environmental monitoring program for clean rooms, or classified environments, involved in the filling and quality control of biological products produced by Butantan Institute, São Paulo, Brazil. This monitoring established the quantification, characterization and seasonality of the microorganisms in air and operators and, moreover, determined the alert and action limits. The total detectable microbial number showed some contrasts in installed air purification systems and in the operational impact on adopted procedures. The typical microbial population consisted of Staphylococcus sp, Micrococcus sp, Bacillus sp and Penicillium sp. The highest microorganism concentration occurred during summer and springtime. The established internal alert and action limits supported the operational procedures. Therefore, the environmental monitoring program is recommended for other laboratories involved in the production of vaccines, hyperimmune sera and biopharmaceuticals.

O presente trabalho teve por objetivo avaliar um programa de monitoramento microbiológico ambiental para áreas limpas, ou ambientes classificados, envolvidas na produção, envasamento e controle dos imunobiológicos produzidos pelo Instituto Butantan. Este monitoramento permitiu a quantificação, a caracterização e a sazonalidade da população microbiana presente no ar e nos operadores, e a determinação dos limites de alerta e ação. O número total de bactérias detectáveis revelou diferenças nos sistemas de purificação de ar instalados e o impacto operacional ocasionado pelos procedimentos realizados. A população microbiana característica foi composta por bactérias dos gêneros Staphylococcus sp, Micrococcus sp, Bacillus sp e por fungos filamentosos do gênero Penicillium sp. A maior concentração de microrganismos ocorreu nos períodos de verão e primavera. Os limites internos de alerta e ação estabelecidos asseguram os procedimentos operacionais, recomendando o monitoramento microbiológico ambiental a outros laboratórios envolvidos na produção de vacinas, soros hiperimunes e imunobiológicos.

Microbiological monitoring of clean rooms in development of vaccines

The aim of the present work was to evaluate an environmental monitoring program for clean rooms, or classified environments, involved in the filling and quality control of biological products produced by Butantan Institute, São Paulo, Brazil. This monitoring established the quantification, characterization and seasonality of the microorganisms in air and operators and, moreover, determined the alert and action limits. The total detectable microbial number showed some contrasts in installed air purification systems and in the operational impact on adopted procedures. The typical microbial population consisted of Staphylococcus sp, Micrococcus sp, Bacillus sp and Penicillium sp. The highest microorganism concentration occurred during summer and springtime. The established internal alert and action limits supported the operational procedures. Therefore, the environmental monitoring program is recommended for other laboratories involved in the production of vaccines, hyperimmune sera and biopharmaceuticals.

O presente trabalho teve por objetivo avaliar um programa de monitoramento microbiológico ambiental para áreas limpas, ou ambientes classificados, envolvidas na produção, envasamento e controle dos imunobiológicos produzidos pelo Instituto Butantan. Este monitoramento permitiu a quantificação, a caracterização e a sazonalidade da população microbiana presente no ar e nos operadores, e a determinação dos limites de alerta e ação. O número total de bactérias detectáveis revelou diferenças nos sistemas de purificação de ar instalados e o impacto operacional ocasionado pelos procedimentos realizados. A população microbiana característica foi composta por bactérias dos gêneros Staphylococcus sp, Micrococcus sp, Bacillus sp e por fungos filamentosos do gênero Penicillium sp. A maior concentração de microrganismos ocorreu nos períodos de verão e primavera. Os limites internos de alerta e ação estabelecidos asseguram os procedimentos operacionais, recomendando o monitoramento microbiológico ambiental a outros laboratórios envolvidos na produção de vacinas, soros hiperimunes e imunobiológicos.