Evaluation of phenotypical and genotypical methods for the identification and typing of Stenotrophomonas maltophilia isolated from a pharmaceutical facility.

AIMS: Evaluate methods for identification and typing of Stenotrophomonas maltophilia isolated from a pharmaceutical facility. METHODS AND RESULTS: From 270 S. maltophilia strains identified by VITEK®2, 40 were selected and submitted to MALDI TOF-MS, 16S and 23S rRNA gene analysis, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), and an antimicrobial susceptibility profile. 16S rRNA sequencing was able to identify 39 (97.5%) strains as Stenotrophomonas spp. and one (2.5%) as Luteimonas huabeiensis. MALDI TOF-MS identified 37 (92.5%) strains as S. maltophilia, and three (7.5%) were not identified. PCR targeting 23S rRNA yielded a positive result for 39 (97.5%) strains. However, after sequencing, two strains were identified as Stenotrophomonas rhizophila, showing false-positive results. The confirmed S. maltophilia strains (n = 37) showed 35 distinct ERIC-PCR profiles and exhibited sensitivity to minocycline and levofloxacin, and six (16.3%) showed intermediate resistance to sulfamethoxazole-trimethoprim. CONCLUSION: Matrix-assisted laser desorption lonization-time of flight mass spectrometry (MALDI-TOF MS) was a satisfactory methodology for the identification of S. maltophilia, but expansion of the database is necessary for the identification of other species. 16S rDNA sequencing showed low resolution for Stenotrophomonas species differentiation. PCR targeting 23S rRNA could not differentiate S. maltophilia from S. rhizophila. ERIC-PCR was shown to be a useful tool for the microbial source tracking of S. maltophilia.

Ensino em tempos de pandemia: adaptações a um grande desafio

Prezados leitores, Desde que recebi o convite para ser editora científica da revista Vigilância Sanitária em Debate: Sociedade, Ciência & Tecnologia (Health Surveillance under Debate: Society, Science & Technology) ­ Visa em Debate, tenho direcionado muita atenção para o fortalecimento da revista como potencial veículo de disseminação de conhecimento em Vigilância Sanitária. As áreas de pesquisa e desenvolvimento tecnológico têm caminhado intrinsecamente, porém outras, como a educação e a formação profissional em Vigilância Sanitária, necessitam de muita promoção e projetos que possibilitem essa formação.

Até quando?

É assustador o cenário que se apresenta, pois vivemos a epidemia dos não vacinados contra a COVID-19. E isso acontece apesar de todas as evidências científicas indicarem que a vacina continua sendo altamente eficaz para a prevenção dos casos graves e que as medidas não farmacológicas (manutenção de distância segura, uso de máscaras e lavagem das mãos) podem proteger a nós mesmos e aos que estão ao nosso redor.

Adaptation and validation of a method for evaluating the bactericidal activity of ethyl alcohol in gel format 70% (w/w).

Ethyl alcohol 70% (w/w) is a broad-spectrum bactericidal agent that is mandatorily marketed as a gel in Brazil since 2002. However, there is no method described for the microbiological quality control of surface disinfectants in the form of gel, which makes it impossible to monitor the effectiveness of these products by the Health Surveillance. Thus, the present study aimed to adapt an analytical method and validate it so that it is possible to evaluate the bactericidal activity of ethyl alcohol-based disinfectants at 70% (w/w) in gel form. The proposed analytical method is an adaptation of a method developed by the Association of Official Analytical Chemists (AOAC), which is currently used to evaluate the bacterial activity of disinfectants in the form of spray and aerosol. The method consisted of challenging the disinfectant by putting it in contact with 60 carriers - previously contaminated with the test microorganism - during the contact time established by the manufacturer. The bacteria used in this work were Staphylococcus aureus CBRVS 00039 ATCC 6538, Salmonella enterica CBRVS 00028 ATCC 10708, and Pseudomonas aeruginosa CBRVS 00025 ATCC 15442. The parameters chosen for the validation of the method were: matrix effect, robustness, and repeatability. The experiments performed with a disinfectant matrix showed that there is no interference of the gel on the effectiveness of alcohol. Satisfactory results regarding the evaluation of bactericidal activity were obtained for the three microorganisms tested when the volume of 100 µL of disinfectant was applied. The method also showed good repeatability, as it proved to be robust with modification of incubation times, equipment and analysts. The development and validation of this method were extremely important for the quality monitoring of these products, besides being the first method described for the evaluation of gel disinfectants.

Polymyxin B Heteroresistance and Adaptive Resistance in Multidrug- and Extremely Drug-Resistant Acinetobacter baumannii.

Acinetobacter baumannii is an emerging pathogen associated with nosocomial infections and multidrug resistance. Polymyxin B has been used to treat infections caused by multidrug-resistant (MDR) A. baumannii but an increase in polymyxin B resistance has been observed. We aimed to determine the diversity, antimicrobial susceptibility, presence of polymyxin B heteroresistance and adaptive resistance in 72 A. baumannii clinical isolates from two public hospitals in Rio de Janeiro. The isolates were identified by sequencing of rpoB gene. Determination of the genetic diversity of isolates was performed by pulsed-field gel electrophoresis and oxacillinases genes were detected by polymerase chain reaction. The polymyxin B heteroresistance was analyzed by population analysis profile and adaptive resistance was evaluated after serial daily passages of isolates in broth containing increasing polymyxin B concentrations. The results showed that 49% of the isolates were collected from respiratory system and 62% were MDR, while 35% were extensively drug resistant. Additionally, all the isolates carried blaOXA-23-like, blaOXA-51-like genes and ISAba1, while 1% had blaOXA-24-like gene. The association of ISAba1-blaOXA-23 was found in 96% of the isolates. Polymyxin B heteroresistance was found in 36% of the isolates and polymyxin B adaptive resistance was not found in the isolates. Our study demonstrated the high resistance to antimicrobials used in clinical practice and the spread of oxacillinases genes and insertion sequence (IS). We also reported the presence of heteroresistance to polymyxin B used as a last-resort therapy for MDR A. baumannii.

Multidrug-resistant Acinetobacter baumannii: differential adherence to HEp-2 and A-549 cells.

Acinetobacter baumannii has been associated with antimicrobial resistance and ability to form biofilms. Furthermore, its adherence to host cells is an important factor to the colonization process. Therefore, this study intended to identify some virulence factors that can explain the success of A. baumannii in causing nosocomial infections. We studied 92 A. baumannii isolates collected from hospitals in Rio de Janeiro, Brazil. Isolates were identified and the susceptibility to antimicrobials was determined. Oxacilinase type ß-lactamase encoding genes were amplified by polymerase chain reaction, and genetic diversity was investigated by pulsed-field gel electrophoresis (PFGE). In addition, biofilm formation on polystyrene plates using crystal violet staining was quantified, and adherence to human cell lines was evaluated. Eighty-six isolates were multidrug-resistant, of which 93% were carbapenem-resistant. All isolates had the blaOXA-51 gene and 94% had the blaOXA-23 gene, other searched blaOXA genes were not detected. PFGE typing showed two predominant clones, and biofilm production was observed in 79% of isolates. A. baumannii isolates adhered better to HEp-2 cell compared with A-549 cell. Clones A, B, E, and F showed a significantly increased adherence to HEp-2 compared with adherence to A-549 cell. Our findings revealed that A. baumannii isolates had high frequencies of resistance to antimicrobial agents, ability to form biofilm, and capacity to adhere to HEp-2 cells.

Assessment of pyrogenic response of lipoteichoic acid by the monocyte activation test and the rabbit pyrogen test.

Lipoteichoic acid (LTA) is a non-endotoxin pyrogen of a great importance in the pathogenesis of sepsis. The Rabbit Pyrogen Test (RPT) is able to detect all types of pyrogens but involves the use of animals. The Bacterial Endotoxin Test (BET) cannot fully replace the RPT because it only detects endotoxins. The Monocyte Activation Test (MAT) is sensitive to all types of pyrogens and it is based on the same biological mechanism that is responsible for the fever reaction in humans. Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) has recommended its use for other pyrogens than endotoxin because its equivalence to RPT can be demonstrated. The aim of this study was to evaluate the pyrogenic responses of the RPT and MAT that was induced by LTA. Different LTA concentrations were assayed by the MAT in parallel to the RPT. The results showed that the MAT was more sensitive than the RPT, demonstrating that the MAT detected LTA. This result may contribute to the acceptance of this test by the Brazilian regulatory agencies as a replacement for the animals used in the RPT.

Avaliação da contaminação microbiana em produtos desinfetantes de uso geral / Microbial contamination in household disinfectant products

Em vista de um estudo desenvolvido no Setor de Saneantes do Instituto Nacional de Controle deQualidade em Saúde da Fundação Oswaldo Cruz, referente à avaliação da atividade antimicrobiana dedesinfetantes, no presente trabalho foram analisados produtos desinfetantes de uso geral disponíveis nomercado brasileiro. Os desinfetantes foram coletados aleatoriamente, dos quais três (produtos A, B e C),apresentaram contaminação microbiana. Para efetuar o isolamento e a identificação dos microrganismoscontaminantes foram utilizados o aparelho Vitek® 2, a amplificação e o sequenciamento do gene rRNA16S. A análise realizada por meio de Vitek® 2 revelou a presença das bactérias Serratia marcescens eAchromobacter xylosoxidans, respectivamente, nos produtos A e B. No produto C foram detectadasAeromonas salmonicida pelo Vitek® 2 e Burkholderia lata pela técnica de amplificação da reação em cadeiada polimerase.

Avaliação dos aspectos do controle da qualidade de produtos cosméticos comercializados no Brasil analisados pelo Instituto Nacional de Controle de Qualidade em Saúde / Analysis on the quality control aspects of cosmetic products commercialized in Brazil performed by the National Institute of Quality Control in Health

Os produtos cosméticos são definidos como preparações constituídas por substâncias naturais ou sintéticas, de uso externo nas diversas partes do corpo humano, que têm o objetivo principal de limpar, perfumar, alterar e/ou corrigir odores corporais e manter o corpo em bom estado. Por ter essas funções, esses produtos são utilizados diariamente por milhões de pessoas – e, por isso, é fundamental efetuar a vigilância da qualidade desses produtos. Neste trabalho, foi examinada a documentação de 133 produtos cosméticos que foram recebidos e analisados no Instituto Nacional de Controle da Qualidade (INCQS) no período de 2005 a 2009. Desvios referentes aos parâmetros físico-químico, microbiológico e toxicológico foram encontrados. O dado mais surpreendente foi referente à rotulagem desses produtos, visto que 94% dos produtos avaliados apresentaram o rótulo em desacordo com a legislação brasileira vigente.

Tolerância de Mycobacterium abscessus subsp. bolletii a desinfetantes de alto nível / Tolerance of Mycobacterium abscessus subsp. bolletii to high-level disinfectants

Mycobacterium abscessus subps. bolletii (clone BRA100), entre outras micobactérias de crescimento rápido (MCR), tem sido isolada como agente etiológico de infecções localizadas e sistêmicas no Brasil. Neste estudo, foi avaliada a suscetibilidade de MCR pelo método confirmatório para Avaliação da Atividade Micobactericida de Desinfetantes frente a produtos à base de glutaraldeído, ácido peracético e ortoftalaldeído. Todos os produtos foram utilizados nas concentrações recomendadas pelos fabricantes.Foram avaliadas cinco cepas clínicas pertencentes ao clone BRA100, juntamente com as cepas de referência M. abscessus ATCC 19977, M. chelonae ATCC 35752, M. fortuitum ATCC 6841 e M. abscessus subps.bolletii INCQS 00594. Os três desinfetantes à base de glutaraldeído não foram eficazes contra M. abscessuse M. abscessus subps. bolletii. Os desinfetantes à base de ácido peracético demonstraram eficácia para todasas micobactérias empregadas, embora as cepas do clone BRA100 tenham demonstrado suscetibilidade reduzida ao ácido peracético B. O produto à base de ortoftalaldeído foi eficaz apenas frente a M. chelonae.Visando a redução de infecções nosocomiais, sugere-se que seja suspenso o uso de produtos à base de glutaraldeído como desinfetante de alto nível para quaisquer finalidades.

Difteria pelo Corynebacterium ulcerans: uma zoonose emergente no Brasil e no mundo / Corynebacterium ulcerans diphtheria: an emerging zoonosis in Brazil and worldwide / Difteria por Corynebacterium ulcerans: una zoonosis emergente en Brasil y en el mundo

O artigo revisa a literatura sobre a emergência de infecções humanas causadas por Corynebacterium ulcerans em diversos países, incluindo o Brasil. Foi realizada análise de artigos publicados entre 1926 e 2011 nas bases Medline/PubMed e SciELO, bem como artigos e informes do Ministério da Saúde. Apresenta-se um esquema de triagem, rápido, econômico e de fácil execução, capaz de permitir a realização do diagnóstico presuntivo de C. ulcerans e C. diphtheriae na maioria dos laboratórios brasileiros públicos e privados. A circulação de C. ulcerans em vários países, aliada aos recentes casos de isolamento do patógeno no Rio de Janeiro, é um alerta a clínicos, veterinários e microbiologistas sobre a ocorrência de difteria zoonótica e a circulação do C. ulcerans em regiões urbanas e rurais do território nacional e/ou da América Latina.

The article is a literature review on the emergence of human infections caused by Corynebacterium ulcerans in many countries including Brazil. Articles in Medline/PubMed and SciELO databases published between 1926 and 2011 were reviewed, as well as articles and reports of the Brazilian Ministry of Health. It is presented a fast, cost-effective and easy to perform screening test for the presumptive diagnosis of C. ulcerans and C. diphtheriae infections in most Brazilian public and private laboratories. C. ulcerans spread in many countries and recent isolation of this pathogen in Rio de Janeiro, southeastern Brazil, is a warning to clinicians, veterinarians, and microbiologists on the occurrence of zoonotic diphtheria and C. ulcerans dissemination in urban and rural areas of Brazil and/or Latin America.

El articulo revisa la literatura sobre la emergencia de infecciones humanas causadas por Corynebacterium ulcerans en diversos países, incluyendo Brasil. Se realizó análisis de artículos publicados entre 1926 y 2011 en las bases Medline/Pubmed y SciELO, así como artículos e informes del Ministerio Brasileño de la Salud. Se presenta un esquema de selección, rápido, económico y de fácil ejecución, capaz de permitir la realización del diagnóstico presuntivo de C. ulcerans y C. diphtheriae en la mayoría de los laboratorios brasileños públicos y privados. La circulación de C. ulcerans en varios países, aliada a los recientes casos de aislamiento del patógeno en Rio de Janeiro (Sureste de Brasil), es un alerta a clínicos, veterinarios y microbiólogos sobre la ocurrencia de difteria zoológica y la circulación de C. ulcerans en regiones urbanas y rurales del territorio nacional y/o de América Latina.

Corynebacterium ulcerans diphtheria: an emerging zoonosis in Brazil and worldwide.

The article is a literature review on the emergence of human infections caused by Corynebacterium ulcerans in many countries including Brazil. Articles in Medline/PubMed and SciELO databases published between 1926 and 2011 were reviewed, as well as articles and reports of the Brazilian Ministry of Health. It is presented a fast, cost-effective and easy to perform screening test for the presumptive diagnosis of C. ulcerans and C. diphtheriae infections in most Brazilian public and private laboratories. C. ulcerans spread in many countries and recent isolation of this pathogen in Rio de Janeiro, southeastern Brazil, is a warning to clinicians, veterinarians, and microbiologists on the occurrence of zoonotic diphtheria and C. ulcerans dissemination in urban and rural areas of Brazil and/or Latin America.

Legislação sanitária brasileira e a comunicação de risco de prudutos de limpeza doméstica / Brazilian sanitary legislation and the risk communication of household cleaning products

It is the duty of the state to protect the population from health risks and one effective measure could be the use of warnings on labels of products available to the general public so that protective measures may be taken. The aim of this study was to analyze the Brazilian sanitary legislation of domestic cleaning products in terms of toxicological information required on the labels. The legislation evaluated referred to antimicrobial, corrosive, pesticide and bleach products in terms of possible health risks. Not all the legislation evaluated required emphasis and definition of appropriate storage; warnings to keep the product out of the reach of children; recommendations to read the instructions before use; the need to keep the product in its original container; the non-reuse of the empty container or a description of the dangers of the product on the labels. The recommendation to take the label to the doctor appeared only under specific legislation; care with kitchen utensils used as a measuring gauge is required only for disinfectants and an emergency phone number appeared in only one law. The results indicate that the regulations for household cleaning products need to be revised in order to make health warnings on the labels more efficient.

É dever do Estado prevenir riscos à saúde e uma medida eficaz poderia ser a comunicação do risco através da rotulagem dos produtos disponíveis à população, para que as pessoas possam tomar medidas de proteção. O objetivo deste estudo foi analisar a legislação sanitária brasileira dos produtos de limpeza domésticos quanto às exigências das informações toxicológicas a serem fornecidas na rotulagem. A legislação avaliada foi a referente aos produtos antimicrobianos, corrosivos, praguicidas e alvejantes, quanto à algumas possíveis situações de risco. Nem todos o documentos avaliados exigem destaque e definem a localização no rótulo para as advertências de manter o produto fora do alcance de crianças, ler as instruções antes de usar, manter o produto na embalagem, não reutilizar a embalagem vazia e para a descrição dos riscos do produto. A recomendação para levar o rótulo ao médico consta apenas na legislação específica, o cuidado com utensílios de cozinha usados como medida é exigido apenas para desinfetantes e, um telefone de emergência é exigido em apenas uma legislação. Os resultados indicam a necessidade de que a legislação que regulamenta os produtos saneantes domisanitários seja revisada tornando acomunicação do risco através da rotulagem mais eficaz.

[Analysis of data from Poison Control Centers in Rio de Janeiro, Brazil, for use in public health activities]. / Análise dos dados dos Centros de Controle de Intoxicação do Rio de Janeiro, Brasil, como subsídio às ações de saúde pública.

Health information systems should be sources of reliable and valid data that can be used for surveillance to reduce morbidity and mortality. Unintentional poisoning is highly relevant to public health, since it is the most frequent cause of emergency pediatric care. Information on such events is recorded in Poison Control Centers (PCCs). The current study analyzed the records on unintentional poisoning with household cleaning products in two PCCs in the State of Rio de Janeiro, Brazil, in 2000-2002. The variables analyzed were gender, age, cause of the accident, exposure route, toxic agent, and outcome. In the 2,810 records studied, boys four years and younger were the most vulnerable group and oral ingestion was the most common exposure route. However, analysis of the data on the toxic agent, cause, and outcome showed that the data currently recorded in the PCCs are inconsistent. It is necessary to review the definitions used by the PCCs in order for the resulting data to effectively support public health measures and health surveillance policies.

Análise dos dados dos Centros de Controle de Intoxicação do Rio de Janeiro, Brasil, como subsídio às ações de saúde pública / Analysis of data from Poison Control Centers in Rio de Janeiro, Brazil, for use in public health activities

Os sistemas de informação em saúde devem ser fonte de informações confiáveis e válidas, para que possam ser utilizadas pelos sistemas de vigilância para reduzir morbidade e mortalidade na população. Intoxicação não intencional é de interesse para a saúde pública, uma vez que é a principal causa de atendimento de emergência pediátrica. As informações sobre estes eventos estão registradas nos Centros de Controle de Intoxicações (CCIs). Este estudo analisou os registros de intoxicações não intencionais com produtos saneantes domissanitários existentes nos dois CCIs do Estado do Rio de Janeiro, Brasil, no período de 2000-2002. As variáveis analisadas foram sexo, idade, causa do acidente, via de exposição, agente tóxico e evolução do caso. Nos 2.810 registros estudados foi identificada a população vulnerável de meninos até quatro anos e a via de exposição (oral). Entretanto, a análise dos dados relativos ao agente tóxico, causa e evolução demonstrou que os dados registrados atualmente nos CCIs são inconsistentes. Há necessidade de revisar as definições adotadas pelos CCIs para que as informações geradas possam ser efetivas em subsidiar ações e políticas de vigilância sanitária.

Health information systems should be sources of reliable and valid data that can be used for surveillance to reduce morbidity and mortality. Unintentional poisoning is highly relevant to public health, since it is the most frequent cause of emergency pediatric care. Information on such events is recorded in Poison Control Centers (PCCs). The current study analyzed the records on unintentional poisoning with household cleaning products in two PCCs in the State of Rio de Janeiro, Brazil, in 2000-2002. The variables analyzed were gender, age, cause of the accident, exposure route, toxic agent, and outcome. In the 2,810 records studied, boys four years and younger were the most vulnerable group and oral ingestion was the most common exposure route. However, analysis of the data on the toxic agent, cause, and outcome showed that the data currently recorded in the PCCs are inconsistent. It is necessary to review the definitions used by the PCCs in order for the resulting data to effectively support public health measures and health surveillance policies.

A profile of unintentional poisoning caused by household cleaning products, disinfectants and pesticides.

Unintentional poisoning occurred mainly among children. The leading cause of such poisoning in Brazil, among consumer products was household cleaning products. For this study 2810 calls made to two poison control centers in the State of Rio de Janeiro between January 2000 and December 2002 were analyzed. Children under five were the most vulnerable group. More boys under 10 suffered accidental poisoning than girls, although above this age, the distribution was inverted. The calls received by poison control centers were mainly from health services within the first two hours following poisoning. The most frequent exposure routes were ingestion (90.4%), followed by inhalation (4.3%), skin and eye contact (2.4% and 2% respectively). The products involved were bleach, petroleum derivates, rodenticides and pesticides. The main causes were products within the children's reach, storage in soft drink bottles, food mixed with rodenticides, incorrect product use, and kitchen utensils used for measured cleaning products. The most common outcome was that the patient was cured, although a lot of cases were lost to follow-up. Education programs are necessary in order to avoid these poisonings.

A profile of unintentional poisoning caused by household cleaning products, disinfectants and pesticides / Perfil de intoxicações não intencionais com produtos saneantes de uso doméstico

Unintentional poisoning occurred mainly among children. The leading cause of such poisoning in Brazil, among consumer products was household cleaning products. For this study 2810 calls made to two poison control centers in the State of Rio de Janeiro between January 2000 and December 2002 were analyzed. Children under five were the most vulnerable group. More boys under 10 suffered accidental poisoning than girls, although above this age, the distribution was inverted. The calls received by poison control centers were mainly from health services within the first two hours following poisoning. The most frequent exposure routes were ingestion (90.4 percent), followed by inhalation (4.3 percent), skin and eye contact (2.4 percent and 2 percent respectively). The products involved were bleach, petroleum derivates, rodenticides and pesticides. The main causes were products within the children's reach, storage in soft drink bottles, food mixed with rodenticides, incorrect product use, and kitchen utensils used for measured cleaning products. The most common outcome was that the patient was cured, although a lot of cases were lost to follow-up. Education programs are necessary in order to avoid these poisonings.

Intoxicações não-intencionais ocorrem principalmente na infância. No Brasil, os produtos de limpeza doméstica são as principais causas desses eventos. Para este estudo foram analisados 2.810 casos registrados nos dois centros de controle de intoxicação do Estado do Rio de Janeiro, no período de janeiro de 2000 a dezembro 2002. O grupo mais vulnerável foi o de crianças de até cinco anos de idade. Meninos de até dez anos foram mais intoxicados do que meninas. A partir dessa faixa etária, a distribuição foi invertida. Os centros de controle de intoxicação foram chamados principalmente por serviços de saúde no período de até duas horas após o envenenamento. As vias de exposição mais freqüentes foram ingestão (90,4 por cento), inalação (4,3 por cento), dérmica (2,4 por cento) e ocular (2 por cento). Os produtos envolvidos foram alvejantes, derivados de petróleo, raticidas e pesticidas. As principais causas foram: produto ao alcance de crianças, estocagem em garrafas de refrigerantes, uso de alimentos com raticidas, uso incorreto do produto, e utensílios de cozinha com produtos de limpeza. O desfecho mais freqüente foi cura apesar do grande número de casos perdidos durante o seguimento. São necessários programas educacionais a fim de evitar esses eventos.