Potentially pathogenic bacteria isolated from neglected air and surfaces in hospitals

Hospital infections (HI) are a serious public health problem in many countries. Several studies have identified strains correlating to surgical site infections, many with multi-resistance. The goals of this study was to quantify, to identify and to verify the resistance profile of microorganisms collected at two hospitals settings, and to alert health professionals how environmental contamination can influence hospital infection rates. For air sampling in operating rooms, intensive care unit and materials sterilization center, the impaction method (Spin Air, IUL®) and passive sedimentation were used. For the isolation of bacteria on surfaces and uniforms contact plates (RODAC®) were used. Identification of the microorganisms was performed using Vitek® 2 Systems. The antibiograms were conducted according to the disk diffusion method recommended by CLSI. The surgical center of hospital B presented more than 500 CFU/m3 in aerial microbial load. In the aerial microbiota of the sampled areas of both hospitals, M. luteus, S. haemolyticus and S. hominis spp hominis were the prevalent microorganisms, with a percentage greater than 30%. On the surfaces and uniforms there was a prevalence of M. luteus (40%) and S. hominis spp hominis (20%) among others, and some of the resistant strains were isolated from environments with microbial load within the recommended limits.

An approach on phytoalexins: function, characterization and biosynthesis in plants of the family Poaceae / Uma abordagem sobre fitoalexinas: função, caracterização e biossíntese em plantas da família Poaceae

ABSTRACT: Phytoalexins are compounds that have been studied a few years ago, which present mainly antimicrobial activity. The plants of the family Poaceae are the most geographically widespread and stand out for their economic importance, once they are cereals used as staple food. This family stands out for having a variety of phytoalexins, which can be synthesized via the shikimic acid (the phenylpropanoids), or mevalonic acid, being considered terpenoid phytoalexins. The characterization of these compounds with antimicrobial activity is carried out using chromatographic techniques, and the high-performance liquid chromatography (HPLC) coupled with mass spectrometry are the most efficient methods in this process. This research aimed to present an approach of the function, characterization and biosynthesis of phytoalexins in plants of the family Poaceae.

RESUMO: As fitoalexinas são compostos que vêm sendo estudados há alguns anos, apresentando principalmente atividade antimicrobiana. As plantas da família Poaceae são as geograficamente mais difundidas e se destacam por sua importância econômica, uma vez que são os cereais que compõe a base da alimentação, em todos os continentes. Essa família se destaca por apresentar uma variedade de fitoalexinas, sendo que as mais importantes são sintetizadas a partir da via do ácido chiquímico. A caracterização desses compostos com atividade antimicrobiana é realizada por meio de técnicas cromatográficas, sendo que a cromatografia líquida de alta eficiência (HPLC) acoplada com espectrometria de massas é a técnica mais eficiente nesse processo. O trabalho apresenta como objetivo uma abordagem da função, caracterização, biossíntese e aplicações das fitoalexinas em plantas da família Poaceae.

Microemulsion based on Pterodon emarginatus oil and its anti-inflammatory potential

This article reports the development of a pharmaceutical product containing vegetable actives from a Brazilian medicinal plant. The possibility of forming a microemulsion using Pterodon emarginatus ("sucupira") oil was evaluated and the anti-inflammatory potential of this microemulsion was also examined. A formulation was developed using P. emarginatus oil, a mixture of ethoxylated Castor Oil (Ultramone(r) R-540/propylene glycol 2:1) (surfactant/cosurfactant) and distilled water at a ratio of 10:15:75, respectively. The microemulsion which was selected was then subjected to the preliminary stability test and analyzed in terms of average diameter of droplets, pH, zeta potential, and polydispersity index, on the 1st, 7th, 15th, and 30th days after preparation and stored at different temperatures (5 ± 2 °C, 25 ± 2 °C, and 40 ± 2 °C). The anti-inflammatory in vivo activity of both oil and formulation were evaluated, using the experimental model of croton oil-induced ear edema. The preliminary stability test showed that the microemulsion stored at 5 and 25 °C retained its original features throughout the 30-day period. The anti-inflammatory potential of both oil and formulation was shown to be statistically significant (p < 0.001), when compared to the control group, however, the microemulsion proved to be more effective (p < 0.05) than the oil when applied directly to the ear.

Descrevemos o desenvolvimento de um produto farmacêutico contendo ativo vegetal baseado em uma planta medicinal brasileira. Foi avaliada a habilidade de formação de uma microemulsão à base do óleo de Pterodon emarginatus (sucupira) e seu potential anti-inflamatório. Uma formulação foi desenvolvida utilizando o óleo de P. emarginatus, mistura de óleo de mamona etoxilado (Ultramona(r) R-540)/propilenoglicol (2:1) (tensoativo/cotensoativo) e água destilada, na proporção de 10:15:75, respectivamente. A microemulsão selecionada foi submetida ao teste preliminar de estabilidade e foi analisada quanto ao diâmetro médio das gotículas, pH, potential zeta e índice de polidispersão, no 1º, 7º, 15º e 30º dias após o preparo e foram estocadas em diferentes temperaturas (5±2 °C, 25±2 °C e 40±2 °C). Avaliaram-se a atividade anti-inflamatória in vivo do óleo de sucupira e da formulação, usando o modelo experimental do edema de orelha induzido pelo óleo de cróton. No teste preliminar de estabilidade observou-se que a formulação estocada a 5 °C e a 25 °C mantiveram suas características originais durante 30 dias. O potencial anti-inflamatório de ambos, óleo de sucupira e formulação foi estatisticamente significativo (p <0.001), quando comparado ao controle, porém a microemulsão foi mais eficiente (p <0.05) que o óleo aplicado diretamente nas orelhas dos animais.

Composição química e atividade antimicrobiana dos óleos essenciais das folhas e flores de Callistemon viminalis (sol. ex Gaertn.) G. Don ex. Loudon (Myrtaceae) / Chemical composition and antimicrobial activity of essential oils of leaves and flowers of Callistemon viminalis (sol. ex Gaertn.) G. Don ex. Loudon (Myrtaceae)

O presente trabalho teve como objetivo determinar a composição química e avaliar a atividade antimicrobiana dos óleos essenciais obtidos das folhas (frescas e secas) e flores da Callistemon viminalis. Os óleos essenciais foram analisados por CG/EM. A concentração inibitória mínima (CIM) foi determinada utilizando o método de microdiluição em caldo frente a bactérias Gram-positivas e Gram-negativas. De um total de 96,72%, 98,70% dos compostos identificados nos óleos essenciais foram monoterpenos. O 1,8-cineol foi o componente majoritário no óleo essencial das folhas frescas (70,43%), folhas secas (71,71%) e flores (42,39%), seguido do tricicleno com 12,97% nas folhas frescas, 8,20% nas folhas secas e 28,08% nas flores. A CIM para todos os micro-organismos testados foi ≥ a 2.000 μg/mL. Esse estudo é o primeiro relato sobre a composição química e atividade antimicrobiana do óleo essencial das flores de C. viminalis.

The aim of this study was to determine the chemical composition and the antimicrobial activity of essential oils of Callistemon viminalis leaves (fresh and dried) and flowers. The essential oils were analyzed by GC/MS and the minimum inhibitory concentration (MIC) against Gram-positive and Gram-negative bacteria was determined by the broth microdilution method. Between 96.72% and 98.70% of the compounds identified in the essential oils were monoterpenes. 1,8-Cineole was the major component in the essential oils of fresh leaves (70.43%), dried leaves (71.71%) and flowers (42.39%), followed by tricyclene: 12.97% in the fresh leaves, 8.20% in the dried leaves and 28.08% in the flowers. The MIC for all microorganisms tested was ≥ 2,000 μg/mL, for all the essential oil samples. This study is the first report on the chemical composition and antimicrobial activity of essential oil from the flowers of C. viminalis.

Composição química e atividade antimicrobiana do óleo essencial e anatomia foliar e caulinar de Citrus limettioides Tanaka (Rutaceae) / Chemical Composition and Antimicrobial Activity of Essential Oil and Anatomy of Citrus limettioides Tanaka(Rutaceae) Leaves and Stem

Citrus limettioides (Rutaceae) é utilizada popularmente para tratamento da sinusite, e também como antitérmica e hipotensora. Este trabalho teve como objetivos: identificar os constituintes dos óleos essenciais da folha e da casca dos frutos de C. limettioides; avaliar a atividade antimicrobiana dos óleos essenciais da folha e da casca dos frutos e realizar o estudo anatômico da folha e caule jovem. Os óleos essenciais foram obtidos por hidrodestilação em aparelho de Clevenger modificado e analisados por CG-EM. A concentração inibitória mínima (CIM) foi determinada utilizando o método de microdiluição em caldo frente a bactérias Gram-positivas, Gram-negativas e fungos. Para o estudo anatômico foram realizados cortes histológicos a mão livre e as secções foram coradas com azul de Alcian/safranina, reagente de Steinmetz, lugol e analisados em microscopia de luz. O linalol foi o componente majoritário no óleo essencial das folhas (36,88 a 45,15%) e o limoneno no óleo da casca dos frutos (70,60 a 75,18%). O óleo essencial das folhas apresentou CIM de 2000 μg/mL frente às bactérias e nenhuma atividade frente aos fungos. O óleo essencial da casca dos frutos não apresentou atividade antimicrobiana. A folha é hipoestomática, predominando os estômatos anomocíticos; o mesofilo é dorsiventral com parênquima paliçádico bisseriado e parênquima lacunoso com 11 camadas de células. Cavidades secretoras foram observadas no mesofilo da folha, no pecíolo e no caule, enquanto cristais prismáticos foram identificados em todas as estruturas da folha e do caule. Os dados obtidos no estudo microscópico podem auxiliar na identificação taxonômica desta planta e na avaliação da autenticidade da droga vegetal.

Citrus limettioides (Rutaceae), a sweet lime, is popularly used to treat sinusitis, and as an antipyretic and hypotensive agent.the aim of this study was to identify the constituents of the essential oils from the leaf and fruit peel of C. limettioides, test the antimicrobial activity of these essential oils and perform an anatomical study of the leaf and young stem. Essential oils were isolated by steam distillation in a modified Clevenger apparatus and analyzed by GC-MS. the minimum inhibitory concentrations (MIC) against Gram-positive and Gram-negative bacteria and fungi were determined by broth microdilution.Anatomical studies were performed on freehand histological sections, which were stained with Alcian blue / safranin, Steinmetz reagent and lugol and analyzed by light microscopy. Linalol was the major component in the leaf essential oil (36.88 to 45.15%) and limonene in the fruit peel essential oil (70.60 to 75.18%).the essential oil from leaves showed antimicrobial activity against bacteria (MIC 2000 μg/mL) and no activity against fungi.the essential oil from the fruit peel showed no antimicrobial activity. the leaf is hypostomatic with predominance of anomocytic stomata, the mesophyll is dorsiventral with biseriate palisade and 11 layers of spongy parenchyma cells.Secretory cavities were observed among the mesophyll cells of the leaf, petiole and stem, whilst prismatic crystals were identified in all structures of leaf and stem. the anatomical data obtained in the microscopic study may help in the taxonomic identification of this plant and to verify the authenticity of the drug plant.

Chemical variability of the essential oils from fruits of Pterodon emarginatus in the Brazilian Cerrado

Pterodon emarginatus Vogel, Fabaceae, is a tree species commonly known as "sucupira-branca". It is a popular medicinal plant in the Brazilian cerrado (Savanna). This study investigates the chemical variability of the essential oils from fruits of P. emarginatus. The fruits were collected from five sites in the Brazilian Cerrado and their essential oils were analyzed by GC/MS. The results obtained by Principal Component and Cluster Analysis identified two groups: cluster I containing β-caryophyllene and δ-elemene and cluster II containing α-copaene, β-cubebene, allo-aromadendrene, α-cubebene and γ-muurulene. The Canonical Discriminant Analysis was used to differentiate between clusters on the basis of essential oil composition. The results showed high variability in the composition of the essential oils from fruits of P. emarginatus, contributing to studies of domestication of this species.

Production and characterization of tyrosinase activity in Pycnoporus sanguineus CCT-4518 crude extract

Tyrosinase is an enzyme of industrial interest. The production and characterization of tyrosinase from P. sanguineus CCT-4518 were investigated. The selection of inductors, luminosity influence, inoculum size and type of culture medium on the production of tyrosinase and the effect of inhibitors on enzyme activity were performed. Optimum conditions for intracellular tyrosinase production was observed after 2 days using 0.15% L-tyrosine as inducer, in the presence of light, with inoculum size of 10 mycelium discs, using 2% malt extract broth medium, incubated at 30°C, and constant agitation of 150 rpm. Tyrosinase activity was completely inhibited by the addition of 6 mM salicylhydroxamic acid or phenylthiourea, however an inhibition of 4.15% was recorded by the addition of 0.1 mM sodium azide. No inhibition could be detected in case of 0.1 mM phenyl methanesulfonyl fluoride addition. Optimal conditions for intracellular tyrosinase activity using L-dopa as substrate were observed at pH 6.6 and 45°C. Thermal stability studies indicated that the enzyme is stable at 45°C for 15 minutes. Higher temperatures decreased tyrosinase activity. Enzyme production was confirmed by non-denaturing polyacrylamide gel electrophoresis and the protein profile was investigated by denaturing polyacrylamide gel electrophoresis.

Dissolution test of herbal medicines containing Paullinia cupana: validation of methods for quantification and assessment of dissolution / Ensaio de dissolução de medicamentos fitoterápicos contendo Paullinia cupana: validação de métodos de quantificação e avaliação de dissolução

"Guaraná" (Paullinia cupana) is used as a physical activity enhancer and stimulator due to its methylxanthines and condensed tannins. The aim of this work was to evaluate the dissolution behavior of five herbal medicines in the form of capsules and tablets containing guaraná. Assay and dissolution methods were validated and results obtained allowed simultaneous marker quantification with precision, accuracy, selectivity and robustness. Findings showed that 100 percent of the herbal medicinal products analyzed did not provide satisfactory results concerning the presence of four markers, 60 percent had three markers (caffeine, catechin and epicatechin), while 40 percent had only caffeine at tested dosage forms. In addition, after 30 minutes, only capsule A showed at least 80 percent of the dissolved markers. In other capsules, marker dissolution did not exceed 60 percent whereas 60 percent of the samples had some characteristic pharmacotechnical problems. These results evidence the need for rigorous quality control to help ensure the therapeutic action of these drugs. To this end, dissolution studies are an essential tool for quality assurance of herbal medicines.

Guaraná (Paullinia cupana) é utilizado como revigorante e estimulante devido à presença de metilxantinas e taninos condensados. Este trabalho visou avaliar o comportamento de dissolução de cinco fitoterápicos, na forma de cápsulas e comprimidos, contendo guaraná. O método de quantificação e de dissolução foram validados e os resultados obtidos permitiram a quantificação dos marcadores simultaneamente, com precisão, exatidão, seletividade e robustez. Foi verificado que 100 por cento dos fitoterápicos analisados encontravam em desacordo quanto à presença dos quatro marcadores, sendo que 60 por cento apresentaram três marcadores (cafeína, catequina e epicatequina) e 40 por cento apresentaram somente a cafeína. Além disso, após o tempo de 30 minutos de ensaio foi possível observar que somente a cápsula A apresentou pelo menos 80 por cento dos marcadores dissolvidos. Nas demais cápsulas o comportamento de dissolução apresentado pelos marcadores não ultrapassou 60 por cento e, além disso, 60 por cento das amostras apresentaram alguns problemas farmacotécnicos característicos. Frente aos resultados obtidos torna-se evidente a necessidade de um rigoroso controle de qualidade que contribua para assegurar a ação terapêutica desses medicamentos e, nesse sentido, o estudo de dissolução constitui uma ferramenta essencial para a garantia de qualidade dos fitoterápicos.

Estudo da atividade antibacteriana de plantas medicinais, aromáticas e corantes naturais / Study of the antibacterial activity of medicinal plants, herbs and natural dyes

O emprego de recursos naturais como mecanismo auxiliar no tratamento de distintas patologias tem ressurgido, com ênfase, nos útimos tempos. Tendo em vista esta situação, este estudo teve por finalidade investigar a atividade antimicrobiana de algumas plantas medicinais e aromáticas bastantes comuns na cultura brasileira. Foi constatado que, de modo particular, o alecrim-pimenta (Lippia sidoides) levou a uma considerável inibição do crescimento bacteriano. Tambêm plantas como alecrim (Rosmarinus officinalis), cravo (Sysygium aromaticum), noz-moscada (Myristica fragrans), pimenta-da-jamaica (Pimenta dioica) e sálvia (Salvia officinalis) demonstraram constituir fontes eficientes de compostos bioativos antibacterianos.

Medicinal plants, natural colour and spices were examined in vitro for antibacterial effects on the growth of Escherichia coli (EPEC) serotype O:125, Salmonella thyphimurium, Staphylococcus aureus, Listeria monocytogenes ATCC 7644 and Yersinia enterocolitica ATCC 9610. The most effective medicinal plant was Lippia sidoides at concentration of 3 percent (w/v), that added to Trypticase Soya Broth completely inhibited the growth of S. thyphimurium, S. aureus, L. monocytogenes and Y. enterocolitica. Among the spices, the cloves (Sysygium aromaticum) extract had the strongest inhibitory activity on the growth of E. coli, S. thyphimurium e Y. enterocolitica demonstred through the agar difussion technique. Sage (Salvia officinalis) and rosemary (Rosmarinus officinalis) extracts inhibited the growth of L. monocytogenes and S. aureus. The nutmeg (Myristica fragrans) extract also prevented the development of L. monocytogenes.