ABSTRACT
A resistência bacteriana tem aumentado progressivamente no mundo, assim, há necessidade de novas opções de tratamentos. A fitoterapia tem ganhado notoriedade para combater infecções, principalmente as causadas por bactérias resistentes aos antibacterianos disponíveis. Diante do exposto, o presente estudo teve como objetivo preparar e analisar a composição fitoquímica e a ação antibacteriana dos extratos hidroetanólicos de canela (EHC) e romã (EHR) isolados e associados frente culturas planctônicas e biofilmes de cepas padrão e clínicas de Acinetobacter baumannii e Pseudomonas aeruginosa, além disso, analisar a ação citotóxica dos extratos em queratinócitos humanos (HaCat). Para isso, os EHC e EHR foram preparados e quantificado o teor de sólidos solúveis. Posteriormente, foi quantificado o teor de flavonoides e fenóis totais, análise antioxidante por meio da redução do radical 2,2'-difenil-1-picrilhidrazila (DPPH), e a fitoquímica por cromatografia líquida (HPLC). Em relação a ação antibacteriana dos extratos, foi aplicado o teste de microdiluição em caldo (CLSI M7-A9) e a ação sinérgica realizada por meio do ensaio de checkerboard. As concentrações mais efetivas foram analisadas sobre biofilmes em formação (prevenção) e biofilmes formados (tratamento de 24 h), e quantificada a viabilidade por meio do teste colorimétrico MTT. Para avaliar a citotoxidade, os tratamentos foram aplicados sobre cultura celular de HaCat por 24 h e analisados por meio do teste colorimétrico MTT. A análise estatística foi realizada com 5% de significância (p<0.05), analisados pelo método ANOVA complementado pelo Teste de Tukey. Os resultados demonstraram que os EHC e EHR possuem ação antioxidante e presença de fitocompostos. Os extratos apresentaram ação antibacteriana para todas as cepas avaliadas, quando os mesmos foram associados, obteve-se concentrações sinérgicas para as cepas clínicas de A. baumannii. Em relação a ação antibiofilme, o EHC inibiu a formação em 95% e EHR em 96% do biofilme de #Ab 1, enquanto a cepa #Pa 2 teve 92% e 93% de inibição quando em contato com EHC e EHR, respectivamente. Após tratamento de 24 h em biofilmes formados, as reduções da viabilidade foram de 72% para as cepas #Ab 2 e #Ab 3 quando em contato com o EHC, já EHR inibiu em 83% a viabilidade da cepa #Ab ATCC. Para P. aeruginosa (#Pa 2), as reduções da viabilidade foram de 84% e 88,5% quando tratados com EHC e EHR, respectivamente. A avaliação da citotoxicidade em HaCat demonstrou que após tratamentos com diferentes concentrações dos extratos a viabilidade celular se manteve acima de 70% em todos os grupos. Diante disso, conclui-se que os EHC e EHR apresentam importante ação antioxidante e antibacteriana, tanto em culturas planctônicas quanto em biofilmes, e não apresentaram efeitos citotóxicos na faixa de concentração testada. (AU)
Bacterial resistance has progressively increased in the world, thus, there is a need for new treatment options. Phytotherapy has gained notoriety for fighting infections, mainly those caused by bacteria resistant to available antibacterials. In view of the above, the present study aimed to prepare and analyze the phytochemical composition and antimicrobial action of hydroethanolic extracts of cinnamon (EHC) and pomegranate (EHR) isolated and associated against planktonic cultures and biofilms of standard and clinical strains of Acinetobacter baumannii and Pseudomonas aeruginosa, in addition, analyze the cytotoxic action of the extracts on human keratinocytes (HaCat). For this, the EHC and EHR were prepared and the soluble solids content was quantified. Subsequently, the content of flavonoids and total phenols, antioxidant analysis through the reduction of the radical 2,2'-diphenyl1-picrylhydrazyl (DPPH), and phytochemistry by liquid chromatography (HPLC) were quantified. Regarding the antimicrobial action of the extracts, the broth microdilution test (CLSI M7-A9) was applied and the synergistic action was performed through the checkerboard test. The most effective concentrations were analyzed on forming biofilms (prevention) and formed biofilms (24 h treatment), and viability was quantified using the MTT colorimetric test. To evaluate the cytotoxicity, the treatments were applied on HaCat cell culture for 24 h and analyzed using the MTT colorimetric test. Statistical analysis was performed with 5% significance (p<0.05), analyzed by the ANOVA method complemented by the Tukey test. The results showed that the EHC and EHR have antioxidant action and presence of phytocompounds. The extracts showed antibacterial action for all evaluated strains, when they were associated, synergistic concentrations were obtained for the clinical strains of A. baumannii. Regarding the antibiofilm action, EHC inhibited formation by 95% and EHR by 96% of the #Ab 1 biofilm, while the #Pa 2 strain had 92% and 93% inhibition when in contact with EHC and EHR, respectively. After 24 h treatment in formed biofilms, viability reductions were 72% for strains #Ab 2 and #Ab 3 when in contact with EHC, whereas EHR inhibited the viability of strain #Ab ATCC by 83%. For P. aeruginosa (#Pa 2), viability reductions were 84% and 88.5% when treated with EHC and EHR, respectively. The evaluation of cytotoxicity in HaCat showed that after treatments with different concentrations of extracts, cell viability remained above 70% in all groups. Therefore, it is concluded that EHC and EHR have important antioxidant and antibacterial action, both in planktonic cultures and in biofilms, and did not show cytotoxic effects in the tested concentration range. (AU)
Subject(s)
Pseudomonas aeruginosa , Cinnamomum zeylanicum , Acinetobacter baumannii , Dental Plaque , Pomegranate , PhytotherapyABSTRACT
BACKGROUND Essential oils (EO) extracted from Cinnamomum verum has been used as an antimicrobial agents for centuries. The effects of C. verum leaf oil against virulence of microorganisms is not well studied yet. OBJECTIVES This study evaluates the effect of C. verum leaf oil against three virulence factors of Candida albicans, C. tropicalis and C. dubliniensis and its in-vivo toxicity. METHODS Chemical composition of EO was determined using gas chromatography-mass spectrometry (GC-MS). Minimum inhibitory concentration (MIC) was determined using clinical and laboratory standards institute (CLSI) M27-A3 broth microdilution. Effect of EO on initial adhesion was quantified using XTT assay after allowing Candida cells to adhere to the polystyrene surface for 2 h. Biofilm formation of Candida in the presence of EO was quantified using XTT viability assay. Efficacy on reduction of germ tube formation was evaluated using standard protocol. Visualisation of biofilm formation and progression under the EO treatment were done using scanning electron microscope (SEM) and Time lapses microscope respectively. In-vivo toxicity of EO was determined using Galleria mellonella larvae. Chlorhexidine digluconate: positive control. RESULTS Eugenol was the main compound of EO. MIC was 1.0 mg/mL. 50% reduction in initial adhesion was achieved by C. albicans, C. tropicalis and C. dubliniensis with 1.0, > 2.0 and 0.34 mg/mL respectively. 0.5 and 1.0 mg/mL significantly inhibit the germ tube formation. MBIC50 for forming biofilms were ≤ 0.35 mg/mL. 1.0 mg/mL prevent biofilm progression of Candida. SEM images exhibited cell wall damages, cellular shrinkages and decreased hyphal formation. No lethal effect was noted with in-vivo experiment model at any concentration tested. CONCLUSION C. verum leaf oil acts against virulence factors of Candida and does not show any toxicity.
Subject(s)
Humans , Candida/drug effects , Oils, Volatile , Cinnamomum zeylanicum/chemistry , Virulence Factors , Antifungal AgentsABSTRACT
Los productos autóctonos, se consideran exóticos por sus materias primas y técnicas particulares de producción y para hacerlos más competitivos en el mercado nacional e internacional, se deben encontrar estrategias de conservación, que prolonguen su vida útil. El propósito de esta investigación fue evaluar la estabilidad química, microbiológica y sensorial del chicheme, elaborado tradicionalmente en el municipio de Ciénaga de Oro, Córdoba Colombia. Se preparó el chicheme con la adición de Syzygium aromaticum (clavo) y Cinnamomum verum (canela), en concentraciones de 730ppm, para el tratamiento T1; 1460ppm, para T2 y 2190ppm, para T3 comparada con un control. También, se efectuó un análisis químico, como pH, acidez y °Brix; además, se realizó un recuento de psicrotróficos, coliformes totales y fecales, bacterias ácido lácticas, Bacilluscereus, Staphylococcus aureus y mohos y levadura. El producto fue envasado en frascos de plástico PET y vidrio de 500mL y almacenados a 4°C, durante 7 días. Los resultados, se sometieron a un análisis de varianza y una prueba de comparación de Tukey, utilizando el paquete estadístico SAS Windows Versión 8. El chicheme adicionado con la concentración más alta de canela y clavo y en envases de vidrio, logró un mayor tiempo de almacenamiento y un menor recuento de mohos y de levaduras. Por el contrario, la bebida colocada en envases de plástico tuvo mayor conteo de microorganismos, debido a que el PET es más poroso que el vidrio.
Local products are considered exotic due to their ingredients and the particular production technics; to make them more competitive in the national and international market, conservation strategies that prolong its shelf life must be found. The purpose of this research was to evaluate the chemical, microbiological and sensory stability of ''chicheme'' prepared traditionally in the municipality of Cienaga del Oro, Cordoba, Colombia. Chicheme was elaborated with the addition of clove Syzygium aromaticum and cinnamon Cinnamomum verumin at concentrations of 730ppm for treatment T1, 1460ppm for T2 and for2190ppm T3 compared with a control. Furthermore, a chemical analysis was performed, determining pH, acidity and °Brix; also a recount of psicrotrofics, total and fecal coliforms, acidic lactic bacteria, Bacillus cereus, Staphylococcus aureus, molds and yeast was carried out. The product was kept in both plastic, PET, and glass jars of 500mL capacity and stored at 4°C for 7 days. The results were subjected to a variance analysis and a Tukey comparison test using the statistical package SAS Windows Version 8. Chicheme added with the highest concentration of cinnamon and cloves and stored in glass containers, showed a longer storage time and a lower count of molds and yeasts. On the other hand, the product placed in plastic containers presented a greater count of microorganisms, probably because PET is more porous than glass.
ABSTRACT
Callosobruchus chinensis (L.) (Coleoptera: Bruchidae) is one of the major insect pest infesting stored pulses in India. Recourse of synthetic insecticides to protect stored pulses often lead to dangers of the development of resistant strains, toxic residue and users safety. Locally available and less toxic pest management alternatives such as the use of effective botanicals are important. Essential oils of two variants of Cinnamomum verum Presl. Syn. C. zelanicum Blume from North East India (RRL J 1620 and RRL J 1622) were characterized and tested for their biological activity in vapour form against Callosobruchus chinensis (L.). Choice and no-choice tests showed that leaf and bark oil of two variants of C. verum Presl. had significant repellent action and reduced the fecundity and decreased egg hatchability of C. chinenisis (L.). However, leaf oil with higher percentage of eugenol exhibited better activity than the bark oil in both the tests. Hence, the volatile essential oils of both the variants of C. verum Presl. can be used safely as fumigants.