The influence of primer choice on archaeal phylogenetic analyses based on 16S rRNA gene PCR / A influência da escolha de iniciadores na filogenia de Archaea em ensaios de PCR para o gene rRNA 16S

Polymerase chain reaction (PCR) assays targeting 16S rRNA genes followed by DNA sequencing are still important tools to characterize microbial communities present in environmental samples. However, despite the crescent number of deposited archaeal DNA sequences in databases, until now we do not have a clear picture of the effectiveness and specificity of the universal primers widely used to describe archaeal communities from different natural habitats. Therefore, in this study, we compared the phylogenetic profile obtained when Cerrado lake sediment DNA samples were submitted to 16S rDNA PCR employing three Archaea-specific primer sets commonly used. Our findings reveal that specificity of primers differed depending on the source of the analyzed DNA. Furthermore, archaeal communities revealed by each primer pair varied greatly, indicating that 16S rRNA gene primer choice affects the community profile obtained, with differences in both taxon detection and operational taxonomic unit (OTU) estimates.

A amplificação de genes que codificam o rRNA 16S por reação em cadeia da polimerase (PCR) e o seu sub sequentesequenciamento consistem em uma ferramenta importante na caracterização de comunidades microbianas presentes em amostras ambientais. No entanto, apesar do crescente número de sequências de DNA de Archaea depositadas em bancos de dados, a especificidade e efetividade dos iniciadores de PCR descritos como universais e amplamente utilizados na descrição desse grupo ainda não está clara. Neste estudo foram comparados os perfis filogenéticos de comunidades de arqueias obtidos a partir amostras de DNA de sedimentos lacustres do Cerrado submetidas a ensaios de PCR empregando três pares de iniciadores específicos para Archaea, comumente utilizados neste tipo de estudo. Nossos resultados indicam que as comunidades de arqueias detectadas com cada par de iniciadores apresentaram grande variação filogenética, sugerindo que a escolha de iniciadores dirigidos ao gene de rRNA 16S tem efeito significativo no perfil da comunidade descrita, com diferenças tanto em relação aos táxons detectados, como nas estimativas de unidades taxonômicas operacionais (OTU).

The influence of primer choice on archaeal phylogenetic analyses based on 16S rRNA gene PCR

Abstract Polymerase chain reaction (PCR) assays targeting 16S rRNA genes followed by DNA sequencing are still important tools to characterize microbial communities present in environmental samples. However, despite the crescent number of deposited archaeal DNA sequences in databases, until now we do not have a clear picture of the effectiveness and specificity of the universal primers widely used to describe archaeal communities from different natural habitats. Therefore, in this study, we compared the phylogenetic profile obtained when Cerrado lake sediment DNA samples were submitted to 16S rDNA PCR employing three Archaea-specific primer sets commonly used. Our findings reveal that specificity of primers differed depending on the source of the analyzed DNA. Furthermore, archaeal communities revealed by each primer pair varied greatly, indicating that 16S rRNA gene primer choice affects the community profile obtained, with differences in both taxon detection and operational taxonomic unit (OTU) estimates.

Resumo A amplificação de genes que codificam o rRNA 16S por reação em cadeia da polimerase (PCR) e o seu subsequente sequenciamento consistem em uma ferramenta importante na caracterização de comunidades microbianas presentes em amostras ambientais. No entanto, apesar do crescente número de sequências de DNA de Archaea depositadas em bancos de dados, a especificidade e efetividade dos iniciadores de PCR descritos como universais e amplamente utilizados na descrição desse grupo ainda não está clara. Neste estudo foram comparados os perfis filogenéticos de comunidades de arqueias obtidos a partir amostras de DNA de sedimentos lacustres do Cerrado submetidas a ensaios de PCR empregando três pares de iniciadores específicos para Archaea, comumente utilizados neste tipo de estudo. Nossos resultados indicam que as comunidades de arqueias detectadas com cada par de iniciadores apresentaram grande variação filogenética, sugerindo que a escolha de iniciadores dirigidos ao gene de rRNA 16S tem efeito significativo no perfil da comunidade descrita, com diferenças tanto em relação aos táxons detectados, como nas estimativas de unidades taxonômicas operacionais (OTU).

The influence of primer choice on archaeal phylogenetic analyses based on 16S rRNA gene PCR / A influência da escolha de iniciadores na filogenia de Archaea em ensaios de PCR para o gene rRNA 16S

Abstract Polymerase chain reaction (PCR) assays targeting 16S rRNA genes followed by DNA sequencing are still important tools to characterize microbial communities present in environmental samples. However, despite the crescent number of deposited archaeal DNA sequences in databases, until now we do not have a clear picture of the effectiveness and specificity of the universal primers widely used to describe archaeal communities from different natural habitats. Therefore, in this study, we compared the phylogenetic profile obtained when Cerrado lake sediment DNA samples were submitted to 16S rDNA PCR employing three Archaea-specific primer sets commonly used. Our findings reveal that specificity of primers differed depending on the source of the analyzed DNA. Furthermore, archaeal communities revealed by each primer pair varied greatly, indicating that 16S rRNA gene primer choice affects the community profile obtained, with differences in both taxon detection and operational taxonomic unit (OTU) estimates.

Resumo A amplificação de genes que codificam o rRNA 16S por reação em cadeia da polimerase (PCR) e o seu subsequente sequenciamento consistem em uma ferramenta importante na caracterização de comunidades microbianas presentes em amostras ambientais. No entanto, apesar do crescente número de sequências de DNA de Archaea depositadas em bancos de dados, a especificidade e efetividade dos iniciadores de PCR descritos como universais e amplamente utilizados na descrição desse grupo ainda não está clara. Neste estudo foram comparados os perfis filogenéticos de comunidades de arqueias obtidos a partir amostras de DNA de sedimentos lacustres do Cerrado submetidas a ensaios de PCR empregando três pares de iniciadores específicos para Archaea, comumente utilizados neste tipo de estudo. Nossos resultados indicam que as comunidades de arqueias detectadas com cada par de iniciadores apresentaram grande variação filogenética, sugerindo que a escolha de iniciadores dirigidos ao gene de rRNA 16S tem efeito significativo no perfil da comunidade descrita, com diferenças tanto em relação aos táxons detectados, como nas estimativas de unidades taxonômicas operacionais (OTU).

Evaluation of an ozone generation system for the control of microorganisms in the indoor air of sanitary environments / Evaluación de un sistema de generación de ozono para el control de microorganismos en el aire interior de ambientes sanitarios

Objetive: Evaluate an Ozone Generating System (OGS) through of the microorganisms count and identificated in the air of the sanitary environments of the National Institute of Health (INS) from Peru. These were The study:exposed to OGS for 1 min per m2, air samples were collected at volumes of 200L, 500Land 1000Lby impaction, pre and post exposure to OGS, cultures were obtained on Plate Count Agar (PCA) and Oxytetracycline Glucose Agar (OGA) grown at 35°C±2°C for 48 hours and 25°C ± 2°C for 5 days.Adecrease Findings:in the count of fungi after exposure to OGS, the difference being statistically significant for the volumes of 200L(p=0.047) and 1000 L(p<0.005); Aspergillus spp. species were identified in most cultures, in bacteria no significant difference was observed.The OGS is a method that inhibits the Conclusions:growth of most fungi in sanitary environments.

Objetivo: Evaluar un Sistema Generador de Ozono (SGO) a través del recuento e identificación de microorganismos en el aire de los ambientes sanitarios del Instituto Nacional de Salud (INS) del Perú. El estudio: Los ambientes fueron expuestos a SGO por 1 min por m2, se recolectaron muestras de aire en volúmenes de 200L, 500L y 1000L por impactación, pre y post exposición a SGO; los cultivos se obtuvieron en Plate Count Agar (PCA) y Oxitetracicline Glucose Agar (OGA) cultivados a 35 °C ± 2 °C durante 48 horas y 25 °C ± 2 °C durante 5 días. Hallazgos: Disminución del recuento de hongos post exposición a SGO, la diferencia fue estadísticamente significativa para los volúmenes de 200L (p=0.047) y 1000 L (p<0.005); especies de Aspergillus spp. fueron identificados en la mayoría de los cultivos; en bacterias no se observaron diferencias significativas. Conclusiones: El SGO es un método que inhibe el crecimiento de la mayoría de hongos en ambientes sanitarios.

Aislamiento microbiológico de superficies inanimadas en contacto con pacientes en un hospital peruano / Microbiological isolation of inanimate surfaces in contact with patients in a peruvian hospital

Resumen Objetivo: describir el perfil microbiológico de las superficies inanimadas en contacto con el paciente en un hospital nivel III de la seguridad social de Chiclayo, Perú. Material y métodos: Se realizó un estudio transversal, con los datos de los informes del Control microbiológico cualitativo de ambientes físicos de 5 servicios de un Hospital de Chiclayo nivel III del Perú. El método para la identificación de microorganismos fue el sistema automatizado VITEK MS. Se presentan análisis des criptivos como frecuencias y porcentajes. Resultados: Se reportaron un total de 177 aislamientos, de los cuales 97,74% (173) fueron positivos, de estos, el 50,87% (88) estuvo conformado por bacilos gram negativos, siendo el microorganismo más aislado Acinetobacter baumannii (17 muestras) seguido de Rhizobium radiobacter (16) y Sphingomonas paucimobilis 13. Conclusiones: El ambiente hospitalario se encuentra altamente contaminado, siendo la mayoría microorganismos patógenos. Estos resultados guardarían relación con el prolongado tiempo de vida de los microorganismos en las superficies inertes y el proceso de limpieza y desinfección del ambiente hospitalario, por lo que la evaluación de su eficacia y el posible desarrollo de nuevas y mejores técnicas de limpieza deben ser motivo de investigación.

Abstract Objective: to describe the microbiological profile of inanimate surfaces in contact with the patient in a social security level III hospital in Chiclayo, Peru. Material and methods: An observational, descriptive, transversal study was carried out with the data from the reports of the Microbiological Qualitative Control of Physical Environments of 5 services of a Chiclayo Hospital level III in Peru. The method for the identification of microorganisms was the automated system VITEK MS. Descriptive analyses such as frequencies and percentages are presented. Results: A total of 177 isolations were reported, from which 97.74% (173) were positive, of these, 50.87% (88) were composed by gram-negative bacilli, being the most isolated microorganism Acinetobacter baumannii (17 samples) followed by Rhizobium radiobacter (16) and Sphingomonas paucimobilis 13. Conclusions: The hospital environment is highly contaminated, being most of them pathogenic microorganisms. These results would be related to the long life of microorganisms on inert surfaces and the process of cleaning and disinfection of the hospital environment, so the evaluation of its effectiveness and the possible development of new and better cleaning techniques should be investigated.

Application of nanopore sequencing in environmental microbiology research / 生物工程学报

The development of high-throughput sequencing techniques enabled a deeper and more comprehensive understanding of environmental microbiology. Specifically, the third-generation sequencing techniques represented by nanopore sequencing have greatly promoted the development of environmental microbiology research due to its advantages such as long sequencing reads, fast sequencing speed, real-time monitoring of sequencing data, and convenient machine carrying, as well as no GC bias and no PCR amplification requirement. This review briefly summarized the technical principle and characteristics of nanopore sequencing, followed by discussing the application of nanopore sequencing techniques in the amplicon sequencing, metagenome sequencing and whole genome sequencing of environmental microorganisms. The advantages and challenges of nanopore sequencing in the application of environmental microbiology research were also analyzed.

Infection risks related to aesthetic procedures: microbial profile and professional perception about infection prevention measures / Riscos de infecção relacionados a procedimentos estéticos: perfil microbiano e percepção profissional sobre medidas de prevenção de infecção / Riesgos de infección relacionados con los procedimientos estéticos: perfil microbiano y percepción profesional sobre las medidas de prevención de infecciones

Background and Objectives: This study aimed to identify the presence of microorganisms in the aesthetic environment and assess professionals' knowledge about relevant infection prevention measures, considering the importance of the issue and the lack of study in the area. Methods: A total of 100 clinics that perform minimally invasive aesthetic procedures in Porto Alegre (RS), Brazil, were visited. Procedures such as botulin-toxin, dermal fillers, collagen biostimulators, thread lift, chemical peels and laser hair removal were considered. A questionnaire about infection prevention measures were answered by 50 professionals. Also, 100 samples were collected from the environment for bacterial identification and antimicrobial susceptibility testing. Results: There was an infection prevention protocol in 40% of clinics, in which 95% of respondents had complete college education. Periodic professional training regarding infection control measures were performed in 72% of clinics. An autoclave was used for sterilization of materials and instruments in 66% of clinics. From the samples collected, 85% showed bacterial growth by microbiological methods. Coagulase-negative Staphylococci was the most prevalent genera found, and 16% of them were resistant to both cefoxitin, erythromycin, and clindamycin. Four isolates were positive for mecA by PCR. Conclusion: The presence of well-trained professionals is critical in aesthetic clinics so that biosafety and infection prevention measures are taken.(AU)

Justificativa e Objetivos: Este estudo teve como objetivo identificar a presença de microrganismos no ambiente estético e avaliar o conhecimento dos profissionais sobre medidas relevantes de prevenção de infecções, considerando a importância do tema e a falta de estudos nesta área. Métodos: Foram visitadas 100 clínicas que realizam procedimentos estéticos minimamente invasivos em Porto Alegre (RS), Brasil. Foram considerados procedimentos injetáveis como aplicação de toxina botulínica, preenchedores faciais, microagulhamento, bioestimuladores de colágeno, fios de sustentação, peelings químicos e depilação a laser. Um questionário sobre medidas de prevenção de infecção foi respondido por 50 profissionais. Além disso, 100 amostras foram coletadas do ambiente para identificação bacteriana e teste de sensibilidade aos antimicrobianos. Resultados: Existia protocolo de prevenção de infecção em 40% dos ambulatórios, no qual 95% dos profissionais entrevistados possuíam ensino superior completo. Treinamento profissional periódico sobre medidas de controle de infecção foi realizado em 72% dos ambulatórios. Autoclave foi utilizada para esterilização de materiais e instrumentais em 66% das clínicas. Das amostras coletadas, 85% apresentaram crescimento bacteriano nas culturas microbiológicas. Staphylococci coagulase-negativo foi o gênero mais prevalente encontrado; e 16% deles eram resistentes à cefoxitina, eritromicina e clindamicina. Quatro isolados foram positivos para mecA por PCR. Conclusão: A presença de profissionais devidamente treinados é fundamental nas clínicas de estética, para que medidas de biossegurança e prevenção de infecções sejam tomadas.(AU)

Justificación y Objetivos: Este estudio tuvo como objetivo identificar la presencia de microorganismos en el entorno estético y evaluar el conocimiento de los profesionales sobre las medidas de prevención de infecciones relevantes, considerando la importancia del tema y la falta de estudios en esta área. Métodos: Se visitaron 100 clínicas que realizan procedimientos estéticos mínimamente invasivos en Porto Alegre (RS), Brasil. Se consideraron procedimientos invasivos, como la aplicación de toxina botulínica, rellenos faciales, microagujas, bioestimuladores de colágeno, hilos de soporte, peelings químicos y depilación láser. Un cuestionario sobre medidas de prevención de infecciones fue respondido por 50 profesionales. Además, se recolectaron 100 muestras del medio ambiente para la identificación bacteriana y las pruebas de susceptibilidad a los antimicrobianos. Resultados: Existía un protocolo de prevención de infecciones en el 40% de las clínicas, en el que el 95% de los profesionales encuestados tenía educación universitaria completa. En el 72% de las clínicas se realizó capacitación profesional periódica sobre medidas de control de infecciones. Se utilizó un autoclave para la esterilización de materiales e instrumentos en el 66% de las clínicas. De las muestras recolectadas, el 85% mostró crecimiento bacteriano por métodos de cultivo microbiologicos. El Staphylococci coagulasa negativo fue el género más prevalente encontrado, y el 16% de ellos eran resistentes tanto a cefoxitina, eritromicina y clindamicina. Cuatro aislamientos fueron positivos para mecA por PCR. Conclusión: La presencia de profesionales debidamente capacitados es fundamental en las clínicas de estética, para la toma medidas de bioseguridad y prevención de infecciones.(AU)

Biological Toxicity Effects of Soil Pollution Caused by Galvanized Wastewater Based on Vibrio Qinghaiensis sp.-Q67 / 法医学杂志

Objective To establish a rapid diagnosis method for the biological toxicity of soil, accurately and rapidly evaluate the toxicity of contaminated sites and identify the dominant pollutants. Methods Take the soil pollution of a galvanized factory as an example, while the metal concentration level was analyzed and detected, a rapid biological toxicity detection method based on the acute toxicity test of luminescent bacteria （Vibrio qinghaiensis sp.-Q67） was established, and the dominant pollutants were identified by stepwise multiple regression. Results The pollutants came from wastewater and metal plating fragments directly discharged from the manufacturing line of the factory. The concentration of those pollutants was correlated with the acute toxicity of Vibrio qinghaiensis sp.-Q67. The dominant pollutants in the study were zinc （Zn）, aluminum （Al） and copper （Cu）. Conclusion The luminescent bacteria toxicity test method based on Vibrio qinghaiensis sp.-Q67 can conveniently and rapidly assess the degree of toxic damage of polluted soil and identify the dominant pollutants and can be applied to the acute toxicity evaluation of polluted soil.

The Novel Coronavirus: An Alert for Pacifiers' Disinfection

Abstract Objective: To evaluate the best strategy for pacifier disinfection methods. Material and Methods: The literature search was conducted on MEDLINE/PubMed, Scielo, Lilacs, Web of Science, and Scopus databases to find all relevant articles published over the past 20 years, based on PRISMA guidelines. Two reviewers extracted data independently by using a standardized form. The following factors were recorded: country of study, type of study, pacifier material, sample number, microorganisms analyzed, decontamination methods used, method accessibility and results found. Results: A total of 121 articles were obtained from all databases. The selected documents underwent a final screening, resulting in 8 articles. The method of disinfection analyzed by the literature were: 3.5% neutral detergent, apple cider vinegar 70% spray, boiling water during 15 minutes, sodium hypochlorite 2.5, hydrogen peroxide 70% spray, chlorhexidine 0.12%, Brushtox®, sterile water and microwave. Conclusion: Because of the broad methods for pacifier disinfection and different levels of accessibility to disinfectant agents, the pacifier consensus for decontamination remains unclear. Although the disinfection methods are diverse, the methods suggested to its disinfection were identified and described in this article.

Aplicaciones biotecnológicas de los microorganismos / Biotechnological applications of microorganisms

Resumen La biodiversidad de los microorganismos así como la naturaleza única y las capacidades biosintéticas en condiciones ambientales específicas hacen que los microorganismos sean los probables candidatos para resolver problemas de escases de alimentos, contro de plagas, biodegradación de los xenobióticos, descomposición de la basura, las pilas de desechos producidas, entre otros. Los microorganismos ofrecen un gran potencial para la exploración de moléculas y procesos, y el conocimiento de las especies no convencionales, especialmente dentro del grupo Archaea, ha estimulado la investigación molecular de genes de interés. Estos nuevos genes pueden incorporarse mediante tecnología recombinante en especies biológicamente conocidas, como E. coli y S. cerevisiae, para la síntesis a gran escala de productos. La microbiología tecnológica tiene grandes potenciales para explorar y obstáculos por superar. Por lo tanto, solo la investigación en esta área resulta prometedora para científicos en todo el mundo. En la presente revisión se presentan las aplicaciones más significativas de los microorganismos en la industria de alimentos, la agricultura, compuestos químicos, combustibles, farmacología y materiales.

Abstract The biodiversity of microorganisms as well as the unique nature and biosynthetic capabilities in specific environmental conditions make microorganisms the likely candidates to solve problems of food shortages, pest control, biodegradation of xenobiotics, decomposition of garbage, batteries of produced waste, among others. Microorganisms offer great potential for the exploration of molecules and processes, and knowledge of non-conventional species, especially within the Archaea group, has stimulated the molecular investigation of genes of interest. These new genes can be incorporated by recombinant technology into biologically known species, such as E. coli and S. cerevisiae, for the large-scale synthesis of products. Technological microbiology has great potentials to explore and obstacles to overcome. Therefore, only research in this area is promising for scientists around the world. In this review we present the most significant applications of microorganisms in the food industry, agriculture, chemical compounds, fuels, pharmacology and materials.

Bioremediation potential of filamentous fungi in methylene blue: Solid and liquid culture media / Potencial de biorremediação de fungos filamentoso em azul de metileno: Meio sólido e líquido

ABSTRACT The evaluation of the bioremediation potential of microbial with dyes in solid and liquid culture media has been described, but prior studies have not mentioned which culture method is most appropriate for selection of microorganisms. Therefore, the aim of this work was to evaluate the bioremediation potential of filamentous fungi in liquid and solid culture media with methylene blue. The fungi isolates tested were Pleurotus ostreatoroseus (POR-SP1, POR-SP2), P. ostreatus (DF39, EF58 and EF60), Pycnoporus sanguineus (PS) and Fusidium sp. (FUS). The methylene blue concentrations tested were 0, 5, 10, and 50 mg L-1 in the solid medium and 0, 5, 25, 50, and 100 mg L-1 in the liquid medium. In the solid medium, the mycelial diameters of DF39, EF58, FUS, and PS were not influenced by the increase in dye concentration. In the liquid medium, DF39, EF58, EF60, and FUS showed a constant methylene blue degradation rate with increasing dye concentration. The dye degradation rate was correlated with the pH of the liquid medium for EF58, EF60, and FUS. The lower diameter growth in the solid medium did not influence the methylene blue dye degradation rate in the liquid medium.

RESUMO A avaliação do potencial biorremediador microbiano com corantes é descrita em meio de cultura sólido e líquido, mas não é mencionado qual método de cultivo é o mais apropriado na seleção de microrganismos. Para tanto, o objetivo deste trabalho foi avaliar o potencial de biorremediação de fungos filamentosos, em meios de cultura líquido e sólido com azul de metileno. Os isolados fúngicos testados foram: Pleurotus ostreatoroseus (POR-SP1 e POR-SP2), P. ostreatus (DF39, EF58 e EF60), Pycnoporus sanguineus (PS) e Fusidium sp. (FUS). As concentrações de azul de metileno testadas foram 0, 5, 10 e 50 mg L-1 em meio sólido e de 0, 5, 25, 50 e 100 mg L-1 em meio líquido. No meio sólido, o diâmetro micelial de DF39, EF58, FUS e PS não foi influenciado pelo aumento da concentração de corante. Em meio líquido, DF39, EF58, EF60 e FUS apresentaram degradação do azul de metileno constante com o aumento da concentração do corante. A taxa de degradação do corante foi correlacionada com o pH do meio líquido, para EF58, EF60 e FUS. O menor crescimento, em diâmetro, em meio sólido, não influenciou na taxa de degradação do corante azul de metileno, em meio líquido.

Análise da qualidade sanitária da areia das praias de Santos, litoral do estado de São Paulo / Analysis of sand quality health in Santos beach, coast of São Paulo state

RESUMO: Em Santos, litoral de São Paulo, a avaliação da qualidade microbiológica da água das praias é realizada pela prefeitura e pela Companhia Ambiental do Estado de São Paulo (CETESB). Entretanto, a mesma atenção não é direcionada à areia das praias. A areia representa um potencial vetor de contaminação e pode constituir reservatório de microrganismos patogênicos. O objetivo deste estudo foi avaliar a qualidade microbiológica da areia das praias de Santos, São Paulo. Utilizou-se a técnica das membranas filtrantes para os grupos de coliformes fecais (Escherichia coli ) e enterococos. As amostras foram coletadas em sete pontos das praias de Santos. Os ensaios foram realizados mensalmente durante 8 meses e os resultados, expressos em unidades formadoras de colônias (UFC) de bactérias por 100 g de areia, variaram de 40.000 a 2.700.000 para E. coli e de não detectado a 95.000 para enterococos, encontrando-se acima dos valores orientadores existentes em âmbito nacional (3.800 UFC.100g-1 - Rio de Janeiro) e internacional (100.000 UFC.100g-1 - Portugal). O contato com areia contaminada pode causar diversas doenças, comprometendo a qualidade de vida da população. Torna-se importante a realização de estudos baseados em evidências epidemiológicas de exposição e análises de risco, para se estabelecer padrões de qualidade e políticas públicas para monitoramento e gerenciamento da qualidade sanitária da areia das praias do litoral de São Paulo.

ABSTRACT: In Santos, coast of São Paulo State, the evaluation of the microbiological quality of the beach water is performed weekly by the Companhia Ambiental do Estado de São Paulo (CETESB), the environmental agency of the São Paulo state government. However, the same attention is not given to the sand of the beaches. Sand represents a potential contamination vector and may constitute a reservoir of pathogenic microorganisms. This study aimed to evaluate the microbiological quality of sands from Santos beaches throughout the quantification of bacteria colonies from the groups of fecal coliforms (Escherichia coli ) and Enterococcus, using the membrane filter technique. The analyses were executed monthly during 8 months and the results, expressed in bacteria colony forming units (CFU) per 100 grams of sand, vary from 40,000 to 2,700,000 for E. coli and from not detected to 95,000 for Enterococcus, being above the currently guideline values in national (3,800 CFU.100g-1) and international (100,000 CFU.100g-1) levels. It's important to mention that these guideline values were based on results from microbiological analyses of sands collected from beaches that are distant from large urban areas. The contact with contaminated sand may cause diseases, provoking impacts on the population's life quality. There is a need for further studies based on epidemiological evidences of exposure and risk analysis, in order to establish quality standards and public policies aimed to monitoring and managing the sanitary quality of sands from São Paulo's coast beaches.

Cryptococcus: isolamento ambiental e caracterização bioquímica / Cryptococcus: environmental isolation and biochemical characterization

O gênero Cryptococcus caracteriza-se por ser uma levedura responsável por infecção sistêmica, causada pelas espécies Cryptococcus neoformans e Cryptococcus gattii. O fungo é encontrado em substratos de origem animal e vegetal, e a infecção ocorre com a inalação de basidiósporos ou leveduras desidratadas infectantes presentes no ambiente. O presente trabalho teve por objetivo pesquisar a existência de microfocos de Cryptococcussp.em amostras ambientais da cidade de Araçatuba, São Paulo, com a finalidade de minimizar os riscos de contaminação do homem e dos animais, buscando o conhecimento da ecoepidemiologia do Cryptococcus. Foram colhidas 50 amostras oriundas de ocos e troncos de árvores (Cassiasp., Ficussp., Caesalpinea peltophorides) de 10 locais representativos do perímetro urbano, as quais foram encaminhadas ao Laboratório de Bacteriologia e Micologia da Faculdade de Medicina Veterinária de Araçatuba-Unesp, onde foram processadas e semeadas em placas de Petri contendo ágar semente de Níger e Sabouraud dextrose com clorafenicol e incubadas à temperatura de 30ºC, por um período não inferior a cinco dias. Posteriormente, foram submetidas às provas bioquímicas: produção de urease, termotolerância a 37ºC e quimiotipagem em ágar CGB (L-canavanina-glicina-azul de bromotimol). A análise dos resultados revelaram que 17 (34%) dos cultivos foram positivos para o gênero Cryptococcus, sendo nove (18%) para Cryptococcus gattiie oito (16%) para Cryptococcus neoformans. Outras leveduras correlacionadas, como Rhodotorula sp. e Candida sp., também foram isoladas. Conclui-se que os basidiósporos de Cryptococcusencontram-se dispersos na natureza, constituindo microfocos ambientais, não vinculados necessariamente a um único hospedeiro.

Cryptococcosis is an opportunistic fungal infection caused by Cryptococcusyeasts, especially C. neoformans and Cryptococcus gattii. The fungus is found in substrates of animal and vegetable origin, and infection occurs through inhalation and seedlings present in the environment. The present study aimed to investigate the existence of microfocus Cryptococcus sp. from the environmental samples of Araçatuba city, São Paulo, featuring new niches, by decoupling the direct relationship between fungus and host in order to minimize the risk of contamination of man and animals, understanding the ecoepidemiology of Cryptococcus. Fifty samples from hollows and tree trunks were harvested (Cassia sp., Ficus sp., Caesalpinea peltophorides) from ten representatives in the urban perimeter. The samples were immediately sent to the Laboratory of Bacteriology and Mycology, Faculty of Veterinary Medicine Araçatuba - Unesp where they were processed and plated on Petri dishes containing agar seed Niger and Sabouraud dextrose agar with chloramphenicol, incubated at 30ºC for a period of no less than 5 days. Afterwards they were subimitted to biochemical tests: urease production, thermotolerance at 37°C and quimiotipagem in CGB agar (L- Canavanine-Glycine-Bromothymol blue). The results showed that 17 (34%) cultures were positive for Cryptococcus, 9 (18%) for Cryptococcus gattii and 8 (16%) for Cryptococcus neoformans. Other yeast correlated as Rhodotorula sp. and Candida sp. were isolated. We conclude that the infectious propagules of Cryptococcus are dispersed in nature and constitute an environmental microfocus, not necessarily being bound to a single host.

Air microorganisms in soldier living quarters of an underground tunnel during air-tight survival training:investigation and analysis / 军事医学

Objective To investigate the distribution of air microorganisms in soldies′living quarters of one under-ground tunnel during airtight survival training .Methods Anderson sampling was carried out in Area A and Area B ( living quarters) and Area C (a toilet) at designated time and place.After sampling, the culture and identification of microorgan-isms were finished .Results ①In the living quarters of the whole underground tunnel , the total number of bacteria ranged from 125 to 37 800/m3,but it was 2692, 1844 and 2199/m3, respectively in Area A, B and C.The mean number of bac-teria was 2245/m3 .The number of fungi ranged from 0 to 10 017/m3 .The total number of fungi of Area A , B and C was 1064 , 883 and 1011/m3 .The mean number was 986/m3 .②The number of bacteria in the living areas presented three fea-tures:the total bacteria showed overall three peaksin Area A and B, buttwin peaksin Area C.In Area A and B, the number of bacteria exhibited low colony at first but fluctuated heavily later .In Area C, it decreased gradually to the mini-mum,and the fungi showed a wavy and irregular trend .③The air microbial species included cocci ( Micrococcus, Coriolis of Staphylococcus aureus, S.epidermidis and S.equorum) , bacilli ( Acinetobacter baumannii, A.lwoffii and Alcaligenes faeca-lis),and fungi (Mucor and Saccharomyces).Conclusion Although the content of microorganisms was up to the military hygiene standards , it was higher than in the same kind of tunnels .Themulti peakphenomenon of microbial distribution suggests that the change of air microorganisms in tunnels has its own characteristics .Most of the air microorganisms are con-ditioned pathogens that may cause illness if they are not under control .

Comparison in disinfection methods and flora detection for object surfaces in laminar flow general intensive care unit / 中国感染控制杂志

Objective To compare the bacteriostasis effect of two disinfectants on frequently touched object sur-faces in laminar flow general intensive care unit (GICU),and investigate bacterial contamination on the object sur-faces,so as to provide reference for proper disinfection method and control of infection.Methods Specimens from surfaces of bed rails and bedside tables were taken for bacterial culture before being disinfected.Then object sur-faces around bed units were disinfected with disinfectant containing 500 mg/L of available chloride (routing group) and GammaTM disinfecting wet wipes (control group)respectively,bacteriostasis rate and qualified rate of bacterial count on object surfaces between two kinds of disinfection methods were compared.Results Before routine disinfec-tion,a total of 87 pathogens were isolated from 200 specimens of object surfaces,bacterial contamination rate was 43.50%. Detection rate of gram-positive bacteria was 78.16%% (n =68),the major were Corynebacterium (26.47%,n = 18), Staphylococcus aureus (23.53%,n=16)and Staphylococcus epidermidis (23.53%,n=16);detection rate of gram-nega-tive bacteria was 21.84%(n=19),the major was Acinetobacter baumannii (63.16%,n=12).After a 10-minute disinfec-tion on surfaces,bacteriostasis rate of routine group and control group was (94.89±7.72)% and(96.33 ±12.88)% re-spectively,there was no significant difference between two groups(P >0.05);qualified rates of bacterial count of object surfaces of two groups were both 100%.Conclusion Regular disinfection of object surfaces around bed units,stand-ardization of the manipulation and hand hygiene compliance of medical personnels are simple and effective method of cutting off bacteria dissemination and preventing healthcare-associated infection.

Evaluation of fecal microorganisms of children with cleft palate before and after palatoplasty

This study isolated and quantified intestinal bacteria of children with cleft palate before and after palatoplasty. A prospective study was conducted from May 2007 to September 2008 on 18 children with cleft palate, aged one to four years, of both genders, attending a tertiary cleft center in Brazil for palatoplasty, to analyze the effect of surgical palate repair on the concentration of anaerobes Bacteroides sp, Bifidobacterium sp and microaerophiles Lactobacillus sp in feces of infants with cleft palate before and 24 hours after treatment with cefazolin for palatoplasty. There was significant reduction of Lactobacillus sp (p < 0.002), Bacteroides sp (p < 0.001) and Bifidobacterium sp (p = 0.021) after palatoplasty, revealing that surgery and utilization of cefazolin significantly influenced the fecal microbiota comparing collections before and after surgery. However, due to study limitations, it was not possible to conclude that other isolated factors, such as surgical stress, anesthetics and other medications used in palatoplasty might have a significant influence on the microbiota. Considering the important participation of the intestinal microbiota on both local and systemic metabolic and immunological activities of the host, professionals should be attentive to the possible influence of these changes in patients submitted to cleft repair.

Innovating Experimental Teaching System and Improving Students’ Practicing Ability——Experimental Teaching Reformation of Environmental Microbiology / 微生物学通报

A new experiment curriculum system of environmental microbiology was established centering on applied microbiology in environmental protection field and emphasizing on design and research experi- ments to motivate the students’ interests for the course, which helped them to improve their ability of think- ing independently and creatively as well as their practicing ability.

The Exploration and Practice of Environmental Microbiology Teaching Reform / 微生物学通报

Environmental Microbiology is an important basic course of Environmental Engineering.Its characteristic is content broad,quick development and strong practicality,thus this curriculum's teaching has certain degree of difficulty.Some suggestions and concrete measures about teaching reform,which included curriculum's course content,teaching method,experiment teaching and assessment methods were proposed in this paper.

Community Structure and Ecological Distribution of Airborne Microbes in Autumn in Xi'an / 环境与健康杂志

Objective To understand the air microorganisms pollution and the community structure and distribution characteristics of outdoor airborne microbes in Xi'an in autumn.Methods Airborne microbes were sampled by fallen plates method in autumn,2006 in Xi'an at three typical sites,factories region,garden greenbelt region and main traffic region.Results The results showed that the total concentration of airborne microbes at the factories region and garden greenbelt region,main traffic region was 2 500 cfu/m~3,1 354 cfu/m~3 and 4 404 cfu/m~3 respectively.Bacteria and fungal genus were predominant.Gram positive bacteria were much more than Gram negative bacteria.There were 20 genera of bacteria and 8 fungal genus were detected. Micrococcui,Bacillus,Staphylococcus and Pseudornonus were predominant.The most frequently isolated fungal genus were Penicillium,Paecilomces,Cladosporium and Nonsporulatiny mycelia.The community structure of airborne fungal genus and the dominant genus in different function region was significantly different.The distribution characteristic of airborne microbe concentration in different function region was different.The concentration of airborne bacteria was significantly lower in garden greenbelt region than factories region and main traffic region,whereas the concentration of airborne fungi in garden greenbelt region was higher than that in factories region and main traffic region.Conclusion The distribution of the airbome microbes shows a characteristic of time and space,it is affected not only by the environment and pollution,but also by the other factors such as human activities,animals and plants.

Distribution Characteristic of Airborne Microbes in Forest Scenic Spot in Huangshan / 环境与健康杂志

Objective To understand the microorganisms pollution and the community structure and distribution characteristics of airborne microbes in forest scenic spot in Huangshan,China.Methods The airborne microbes were sampled by fallen plate method from 2002 to 2003 in Huangshan scenic area and Hefei city.Results The results showed that there was a significant negative correlation between the elevation and airborne microbe,the relative coefficient was -0.580(P