Influence of glucose supplementation on biofilm formation of Candida albicans and Candida glabrata isolated from diabetic and non-diabetic individuals.

OBJECTIVE: This study evaluated the effect of different glucose concentration on biofilm formation of Candida albicans and Candida glabrata strains isolated from diabetic and non-diabetic individuals. METHODS: The study was divided into two stages: (I) selection and identification of 48 C. albicans and C. glabrata strains by polymerase chain reaction followed by restriction fragment length polymorphism analysis (PCR/RFLP); (II) evaluation of biofilm formation by means of viability rates (colony-forming units), biofilm dry matter (mg) and biofilm-covered areas (µm2). Statistical comparisons were performed through nonparametric analysis of longitudinal data in factorial experiments with pairwise comparisons using Friedman Conover's test (α = 0.05). RESULTS: All the Candida spp. had their identifications confirmed by PCR/RFLP. C. albicans biofilm of strains from diabetic individuals cultivated in different glucose concentration showed higher viability rates than strains from non-diabetic individuals. No difference was observed on viability of C. glabrata biofilm. Regarding biofilm dry matter, C. albicans biofilm of strains from diabetic individuals cultivated in different glucose concentration showed lower amount in weight than strains from non-diabetic individuals. In C. glabrata strains, this result was only observed in biofilms cultivated with no glucose supplementation. With regard to biofilm-covered areas, only glucose supplementation and non-diabetic condition showed a positive effect on C. albicans biofilm development, and no condition affected C. glabrata biofilm formation. CONCLUSION: The strain type (C. albicans and C. glabrata) isolated from diabetic and non-diabetic individuals influenced on biofilm formation, but glucose supplementation did not.

Avaliação in vitro da formação de biofilme por cepas de Candida spp. provenientes de indivíduos diabéticos e não diabéticos em ambiente sem e com suplementação de glicose / In vitro evaluation of biofilm formation by Candida spp. strains from diabetic and non-diabetic individuals in environment without and with glucose supplementation

É notório o papel do diabetes mellitus como fator de risco para o desenvolvimento de doenças bucais, como a candidíase bucal, cárie e a periodontite. Assim, este estudo tem como objetivo avaliar a formação de biofilme por cepas de Candida spp. provenientes de diabéticos e não diabéticos em ambiente sem e com suplementação de glicose. Trata-se de um estudo experimental laboratorial in vitro, em três etapas. Etapas um e dois, de obtenção e identificação de 48 cepas de Candida spp., sendo que 32 de C. albicans e 16 de C. glabrata, com auxílio da técnica de PCR. Ainda, a etapa três, de processamento microbiológico, com a avaliação da capacidade de formação de biofilme por três ensaios distintos: I) determinação do número de unidades formadoras de colônia (UFC/mL); II) matéria seca dos biofilmes; III) taxa de crescimento de biofilme em fundo de placa de poliestireno. Inicialmente, objetivando simular as características observadas in vivo, o fundo das placas de cultivo recebeu 400 µL de saliva humana para formação da película adquirida. Decorrida a incubação a 37 °C por 24 h, a saliva foi descartada e cada poço de cultura recebeu suspensão padronizada das leveduras (106 UFC/mL) em Saubouraud Dextrose Broth sem suplementação e com suplementação de glicose a 2 e 10 mg/mL, e as placas foram incubadas a 37 °C por 48 h. Para avaliação do número de UFC/mL, o biofilme aderido foi coletado, diluído seriamente e cultivado em placas de Petri com Sabouraud Dextrose Agar. Após incubação os resultados foram expressos em log UFC/mL. Para a avaliação da matéria seca, a solução remanescente foi liofilizada e mensurada em balança de precisão. A taxa de crescimento de biofilme foi avaliada por microscopia Operetta CLS High Content e o FilmTracer(TM) LIVE/DEAD Biofilm Viability kit, conforme o protocolo do fabricante. Posteriormente, 10 imagens por poço foram obtidas e digitalizadas com ampliação de 40 ×. A área recoberta por biofilme (µm2) das imagens foi avaliada com auxílio do software Harmony High Content Imaging. Os dados apresentaram distribuição não normal, e a comparação entre as cepas de diabéticos e não diabéticos foi realizada pelo teste U Mann-Whitney. O teste de Kruskal-Wallis one way foi utilizado para verificar diferenças entre as condições de suplementação de glicose. O nível de significância estatística adotado foi de α = 5%. Os valores de UFC/mL mostraram um maior crescimento das cepas de C. albicans dos pacientes diabéticos em relação aos não diabéticos nas três suplementações (p < 0,001). Por outro lado, acerca da matéria seca em 10 mg/mL e da taxa de crescimento de biofilme sem suplementação de glicose e a 2 mg/mL, os resultados indicaram uma formação de biofilme maior para cepas de C. albicans dos não diabéticos (p < 0,001). Em conclusão, cepas de C. albicans e C. glabrata provenientes de diabéticos e não diabéticos em ambiente sem e com suplementação de glicose apresentaram resultados distintos quanto à formação de biofilme, por diferentes técnicas

The role of diabetes mellitus is notorious as a risk factor for development of oral diseases, such as oral candidiasis, dental caries, and periodontitis. Thus, this study aimed to evaluate biofilm formation by Candida spp. strains from diabetic and non-diabetic individuals in environment without and with glucose supplementation. This is an in vitro experimental laboratory study, in three stages. Stages one and two of obtainment and identification of 48 Candida spp. strains, with 32 of C. albicans and 16 of C. glabrata, with the help of PCR technique. Also, stage three, of microbiological processing, with evaluation of biofilm formation capacity by three different assays: I) determination of the number of colony forming units (CFU/mL); II) biofilm dry matter; III) biofilm growth rate on the bottom of polystyrene plates. Initially, aiming to simulate the characteristics observed in vivo, the bottom of the cultivation plates received 400 µL of human saliva for formation of acquired pellicle. After the incubation at 37 °C for 24 h, the saliva was discarded and each culture well received standardized suspension of yeast (106 CFU/mL) in Saubouraud Dextrose Broth without supplementation and with glucose supplementation at 2 and 10 mg/mL, and the plates were incubated at 37 °C for 48 h. To assess the number of CFU/mL, the adhered biofilm was collected, seriously diluted, and cultivated in Petri dishes with Sabouraud Dextrose Agar. After the incubation, the results were expressed in log CFU/mL. To assess the dry matter, the remaining solution was lyophilized and measured on a precision scale. The biofilm growth rate was evaluated by Operetta CLS High Content microscopy and FilmTracer(TM) LIVE/DEAD Biofilm Viability kit, according to manufacturer's protocol. Later, 10 images per well were obtained and digitalized with 40 × magnification. The area covered by biofilm (µm2) of the images was assessed with the help of Harmony High Content Imaging software. Data showed non-normal distribution, and the comparison among the diabetic and non-diabetic strains was performed by Mann-Whitney U test. Kruskal-Wallis one-way test was used to verify differences between conditions of glucose supplementation. The level of statistical significance adopted was α = 5%. The values of CFU/mL showed greater growth of the diabetic patient's strains in relation to the non-diabetic ones (p < 0.001). On the other hand, regarding dry matter at 10 mg/mL and the growth rate of biofilm without glucose supplementation and at 2 mg/mL, the results indicated a higher biofilm formation for strains of C. albicans from non-diabetic individuals (p <0.001). In conclusion, C. albicans and C. glabrata strains from diabetic and non-diabetic individuals in environment without and with glucose supplementation showed different results concerning the biofilm formation, using different techniques

Patient safety and infection control: bases for curricular integration.

OBJECTIVE: To analyze curricular integration between teaching of patient safety and good infection prevention and control practices. METHOD: Integrative review, designed to answer the question: "How does curricular integration of content about 'patient safety teaching' and content about 'infection prevention and control practices' occur in undergraduate courses in the health field?". The following databases were searched for primary studies: CINAHL, LILACS, ScienceDirect, Web of Science, Scopus, Europe PMC and MEDLINE. RESULTS: The final sample consisted of 13 studies. After content analysis, primary studies were grouped into two subject categories: "Innovative teaching practices" and "Curricular evaluation. CONCLUSION: Patient safety related to infection prevention and control practices is present in the curriculum of health undergraduate courses, but is not coordinated with other themes, is taught sporadically, and focuses mainly on hand hygiene.

Patient safety and infection control: bases for curricular integration / Seguridad del paciente y control de infección: bases para la integración curricular / Segurança do paciente e controle de infecção: bases para a integração curricular

ABSTRACT Objective: To analyze curricular integration between teaching of patient safety and good infection prevention and control practices. Method: Integrative review, designed to answer the question: "How does curricular integration of content about 'patient safety teaching' and content about 'infection prevention and control practices' occur in undergraduate courses in the health field?". The following databases were searched for primary studies: CINAHL, LILACS, ScienceDirect, Web of Science, Scopus, Europe PMC and MEDLINE. Results: The final sample consisted of 13 studies. After content analysis, primary studies were grouped into two subject categories: "Innovative teaching practices" and "Curricular evaluation. Conclusion: Patient safety related to infection prevention and control practices is present in the curriculum of health undergraduate courses, but is not coordinated with other themes, is taught sporadically, and focuses mainly on hand hygiene.

RESUMEN Objetivo: Analizar, partiendo de la literatura, la integración curricular entre enseñanza sobre seguridad del paciente y buenas prácticas de prevención y control de infección. Método: Revisión integrativa orientada a responder la pregunta: "¿Cómo sucede la integración curricular entre los contenidos 'enseñanza sobre seguridad del paciente' y 'prácticas de prevención y control de infección' en cursos de grado en salud?". Se utilizaron las bases CINAHL, LILACS, Science Direct, Web of Science, SCOPUS, Europe PMC y Medline para recuperar estudios primarios. Resultados: Muestra compuesta por trece estudios. Luego de análisis de contenido, los estudios primarios fueron subdivididos en dos categorías temáticas: "Prácticas de enseñanza innovadoras" y "Evaluación Curricular". Conclusión: La seguridad del paciente respecto de prácticas de prevención y control de infección, a pesar de contemplarse en los currículos de cursos de grado en salud, es enseñada de modo puntual, diluido y desarticulado, con foco predominante en la higiene de las manos.

RESUMO Objetivo: Analisar, a partir da literatura, a integração curricular entre o ensino da segurança do paciente e as boas práticas de prevenção e o controle de infecção. Método: Revisão integrativa, com vistas à responder a seguinte questão: "Como ocorre a integração curricular entre os conteúdos 'ensino da segurança do paciente' e 'práticas de prevenção e controle de infecção' nos cursos de graduação em saúde?". Utilizou-se as bases CINAHL, LILACS, Science direct, Web of Science, SCOPUS, Europe PMC, e Medline para recuperar os estudos primários. Resultados: 13 estudos compuseram a amostra. Após uma análise de conteúdo, os estudos primários foram subdivididos em duas categorias temáticas: "Práticas de ensino inovadoras" e "Avaliação Curricular". Conclusão: A segurança do paciente, relacionada às práticas de prevenção e controle da infecção, embora contemplada nos currículos de ensino de graduação em saúde, é ensinada de forma pontual, diluída e desarticulada, focadas predominantemente na higiene das mãos.