AVALIAÇÃO DA VIABILIDADE DE BACTÉRIAS ESTOCADAS EM CALDO BHI-GLICEROL A -20ºC HÁ QUATRO ANOS

With the biotechnological and scientific advancements, the need for preserving microorganisms in laboratories for diagnostic, research, or educational purposes arises. Thus, this study aims to analyze the viability of bacteria stored for four years using the cryopreservation technique. The viability of 70 bacterial isolates was evaluated, including 17 species of Gram-positive bacteria and 53 species of Gram-negative bacteria stored in cryogenic tubes containing Brain Heart Infusion (BHI) broth and glycerol, maintained in a -20ºC freezer since 2019. To reactivate the isolates, bacterial suspensions were thawed and plated on Mueller Hinton Agar plates, while the remaining material was added to tubes containing BHI broth and incubated in a bacteriological incubator at 35ºC for 24 hours. The results demonstrated that 84.3% (59) of the samples showed satisfactory growth without contamination, with Gram-negative bacteria achieving a higher viability rate (84.9% - 45) compared to Gram-positive bacteria (82.3% - 14). Therefore, this study demonstrates that freezing bacteria in BHI-glycerol broth at -20ºC is an effective technique for the long-term conservation of Gram-negative bacteria.Con el desarrollo biotecnológico y científico, existe la necesidad de preservar los microorganismos en el laboratorio con fines de diagnóstico, investigación o enseñanza. Por lo tanto, este trabajo tiene como objetivo analizar la viabilidad de las bacterias almacenadas durante cuatro años, utilizando la técnica de criopreservación. Se evaluó la viabilidad de 70 aislados bacterianos, 17 especies de bacterias Gram-positivas y 53 bacterias Gram-negativas, almacenadas en tubos criogénicos que contienen caldo de Infusión Cerebral Corazón (IHB) y glicerol mantenido en un congelador a -20ºC desde 2019. Para reactivar los aislamientos, las suspensiones bacterianas se descongelaron y se sembraron en placas de Agar Mueller Hinton y el resto del material en tubos que contenían caldo BHI y se incubaron en un horno bacteriológico a 35ºC durante 24 horas. Los resultados mostraron que el 84,3% (59) de las muestras mostraron un crecimiento satisfactorio y ninguna contaminación, y las bacterias Gram-negativas alcanzaron la mayor tasa de viabilidad (84,9% - 45) en comparación con las Gram-positivas (82,3% - 14). Así, el presente estudio demostró que la congelación de bacterias en caldo de BHI-glicerol a -20ºC es una técnica eficaz para la conservación de bacterias Gram-negativas durante largos períodos.Com o desenvolvimento biotecnológico e científico, surge a necessidade da preservação de micro-organismos em laboratório para fins de diagnóstico, pesquisa ou ensino. Dessa forma, este trabalho objetiva analisar a viabilidade de bactérias estocadas há quatro anos, utilizando a técnica de criopreservação. Foi avaliada a viabilidade de 70 isolados bacterianos, sendo 17 espécies de bactérias Gram positivas e 53 bactérias Gram negativas, estocadas em tubos criogênicos contendo caldo Brain Heart Infusion (BHI) e glicerol mantidos em freezer a -20ºC desde 2019. Para reativação dos isolados, as suspensões bacterianas foram descongeladas e semeadas em placas de Ágar Mueller Hinton e o restante do material em tubos contendo caldo BHI e incubado em estufa bacteriológica a 35ºC por 24 horas. Os Resultados demonstraram que 84,3% (59) das amostras apresentaram crescimento satisfatório e sem contaminação, sendo que as bactérias Gram negativas alcançaram a maior taxa de viabilidade (84,9% - 45) em comparação com as Gram positivas (82,3% - 14). Sendo assim, o presente estudo demonstrou que o congelamento de bactérias em caldo BHI-glicerol a -20ºC é uma técnica eficaz para a conservação de bactérias Gram negativas por longos períodos.Com o desenvolvimento biotecnológico e científico, surge a necessidade da preservação de micro-organismos em laboratório para fins de diagnóstico, pesquisa ou ensino. Dessa forma, este trabalho objetiva analisar a viabilidade de bactérias estocadas há quatro anos, utilizando a técnica de criopreservação. Foi avaliada a viabilidade de 70 isolados bacterianos, sendo 17 espécies de bactérias Gram positivas e 53 bactérias Gram negativas, estocadas em tubos criogênicos contendo caldo Brain Heart Infusion (BHI) e glicerol mantidos em freezer a -20ºC desde 2019. Para reativação dos isolados, as suspensões bacterianas foram descongeladas e semeadas em placas de Ágar Mueller Hinton e o restante do material em tubos contendo caldo BHI e incubado em estufa bacteriológica a 35ºC por 24 horas. Os Resultados demonstraram que 84,3% (59) das amostras apresentaram crescimento satisfatório e sem contaminação, sendo que as bactérias Gram negativas alcançaram a maior taxa de viabilidade (84,9% - 45) em comparação com as Gram positivas (82,3% - 14). Sendo assim, o presente estudo demonstrou que o congelamento de bactérias em caldo BHI-glicerol a -20ºC é uma técnica eficaz para a conservação de bactérias Gram negativas por longos períodos

44(5)2011.indd

ABSTRACT Introduction: Melanin production by species of Cryptococcus is widely used to characterize C. neoformans complex in mycology laboratories. This study aims to test the efficacy of methyldopa from pharmaceutical tablet as a substrate for melanin production, to compare the production of melanin using different agar base added with methyldopa, and to compare the melanin produced in those media with that produced in Niger seed agar and sunflower seed agar by C. neoformans, C. laurentii, and C. albidus. Two isolates of each species, C. neoformans, C. laurentii, and C. albidus, and one of Candida albicans were used to experimentally detect conditions for melanin production. Methods: The following media were tested: MuellerHinton agar (MHA), brain and heart infusion agar (BHIA), blood agar base (BAB), and minimal medium agar (MMA), all added with methyldopa, and the media Niger seed agar (NSA) and sunflower seed agar (SSA). Results: All isolates grew in most of the culture media after 24h. Strains planted on media BAB and BHIA showed growth only after 48h. All isolates produced melanin in MMA, MHA, SSA, and NSA media. Conclusions: Methyldopa in the form pharmaceutical tablet can be used as a substrate for melanin production by Cryptococcus species; minimal medium plus methyldopa was more efficient than the BAB, MHA, and BHIA in the melanin production; and NSA and SSA, followed by MMA added with methyldopa, were more efficient than other media studied for melanin production by all strains studied

Prevalence of intestinal parasites in preschool children in the region of Uberlândia, State of Minas Gerais, Brazil Prevalência de parasitoses intestinais em crianças institucionalizadas na região de Uberlândia, Estado de Minas Gerais

INTRODUCTION: Children are an important high-risk group for helminth and protozoa infections. Daycare centers are environments where children have proven to be more susceptible to acquiring intestinal parasites. Thus, the purpose of this study was to verify the prevalence of intestinal parasites in children who attended the two daycare centers maintained by the local government of Uberlândia, State of Minas Gerais, Brazil. METHODS: Fecal samples were collected from 133 children (73 children at the Public Preschool for Early Childhood Education, PPECE A, and 60 at the PPECE B) following identification according to sex and age and agreement to participate by parents or guardians who signed the free, informed consent form. The samples were examined by the Lutz method. RESULTS: Coproparasitological tests performed on 133 children showed that 29.3% of them were parasitized for enteroparasites or commensals, 6.7% of the children presented polyparasitism. Among the protozoa, Giardia lamblia were the most prevalent and Hymenolepis nana were the most frequent among the helminths. CONCLUSIONS: Thus, analysis of the results showed that intestinal parasites still represent a public health problem, especially among children and in areas where the socioeconomic and educational conditions are less favorable.<br>INTRODUÇÃO: As crianças são importantes grupos de risco para infecções por helmintos e protozoários. Os centros de educação infantil são ambientes onde as crianças estão mais expostas à infecção por parasitas intestinais. O objetivo deste estudo foi verificar a prevalência de parasitas intestinais em crianças de duas creches mantidas pelo governo municipal de Uberlândia, Minas Gerais, Brasil. MÉTODOS: Os exames coproparasitológicos foram realizados em 133 crianças (73 crianças da Escola Municipal de Educação Infantil - EMEI A e 60 da EMEI B), depois da identificação da criança de acordo com a idade e sexo, e concordância dos responsáveis através do Termo de Consentimento Livre e Esclarecido. As amostras foram examinadas pelo método de Lutz. RESULTADOS: Os exames coproparasitológicos demonstraram que 29,3% delas estavam parasitadas por enteroparasitos ou comensais e 6,7% das crianças apresentaram poliparasitismo. Entre os protozoários, Giardia lamblia foi o mais prevalente, enquanto Hymenolepis nana foi mais frequente dentre os helmintos. CONCLUSÕES: Assim, podemos observar que as parasitoses intestinais ainda representam um problema de saúde pública, especialmente entre as crianças e em áreas onde as condições socioeconômicas e educacionais são menos favoráveis

Prevalence of intestinal parasites in preschool children in the region of Uberlândia, State of Minas Gerais, Brazil

INTRODUCTION: Children are an important high-risk group for helminth and protozoa infections. Daycare centers are environments where children have proven to be more susceptible to acquiring intestinal parasites. Thus, the purpose of this study was to verify the prevalence of intestinal parasites in children who attended the two daycare centers maintained by the local government of Uberlândia, State of Minas Gerais, Brazil. METHODS: Fecal samples were collected from 133 children (73 children at the Public Preschool for Early Childhood Education, PPECE A, and 60 at the PPECE B) following identification according to sex and age and agreement to participate by parents or guardians who signed the free, informed consent form. The samples were examined by the Lutz method. RESULTS: Coproparasitological tests performed on 133 children showed that 29.3% of them were parasitized for enteroparasites or commensals, 6.7% of the children presented polyparasitism. Among the protozoa, Giardia lamblia were the most prevalent and Hymenolepis nana were the most frequent among the helminths. CONCLUSIONS: Thus, analysis of the results showed that intestinal parasites still represent a public health problem, especially among children and in areas where the socioeconomic and educational conditions are less favorable

Inanimate Surfaces and Air Contamination with Multidrug Resistant Species of Staphylococcus in the Neonatal Intensive Care Unit Environment

Background: Contamination of the hospital environment with multi-resistant (MDR) Staphylococcus increases the risk of infection. The aim of this study is to identify the MDR species of Staphylococcus on inanimate surfaces, in air, and in clinical samples, and analyze the risk factors that correlate with the occurrence of infections in a Neonatal Intensive Care Unit. Methods: Samples of inanimate surfaces and air were taken using a premoistened swab (0.9% sodium chloride) and spontaneous air sedimentation, respectively. The clinical isolates were recovered from infected neonates. The isolates (environmental and clinical) were identified by matrix-assisted laser desorption ionization-time of flight and the resistance profile was calculated using the disk diffusion agar technique. Results: In total, 181 isolates were obtained, 93 from (surfaces), 18 from the air, and 70 clinical samples. S. epidermidis was the most frequent species (66.8%), and the failure rate in air cleaning was 100%. More than 60% of the isolates were MDR, and the majority of clinical isolates (60.4%) had a resistance profile identical to that of the environmental isolates. Conclusion: Staphylococcus spp. were found in most of the analyzed samples, with a high frequency of MDR isolates, demonstrating the importance of the hospital environment as a reservoir, and the need for infection control measures, and rational use of antimicrobials

Different culture media containing methyldopa for melanin production by Cryptococcus species

INTRODUCTION: Melanin production by species of Cryptococcus is widely used to characterize C. neoformans complex in mycology laboratories. This study aims to test the efficacy of methyldopa from pharmaceutical tablet as a substrate for melanin production, to compare the production of melanin using different agar base added with methyldopa, and to compare the melanin produced in those media with that produced in Niger seed agar and sunflower seed agar by C. neoformans, C. laurentii, and C. albidus. Two isolates of each species, C. neoformans, C. laurentii, and C. albidus, and one of Candida albicans were used to experimentally detect conditions for melanin production. METHODS: The following media were tested: Mueller-Hinton agar (MHA), brain and heart infusion agar (BHIA), blood agar base (BAB), and minimal medium agar (MMA), all added with methyldopa, and the media Niger seed agar (NSA) and sunflower seed agar (SSA). RESULTS: All isolates grew in most of the culture media after 24h. Strains planted on media BAB and BHIA showed growth only after 48h. All isolates produced melanin in MMA, MHA, SSA, and NSA media. CONCLUSIONS: Methyldopa in the form pharmaceutical tablet can be used as a substrate for melanin production by Cryptococcus species; minimal medium plus methyldopa was more efficient than the BAB, MHA, and BHIA in the melanin production; and NSA and SSA, followed by MMA added with methyldopa, were more efficient than other media studied for melanin production by all strains studied