RESUMO
Resumen La clorhexidina como tratamiento de la enfermedad periodontal ha logrado efectos bactericidas sobre periodontopatógenos y biopelícula oral. Su uso genera efectos adversos, por lo tanto se presentan alternativas naturales con efecto antimicrobiano similar. Los aceites esenciales han demostrado efectividad en el control de la placa dental, sin los efectos adversos de la clorhexidina. El objetivo de este estudio fue determinar el efecto bacteriostático y bactericida del aceite esencial de mandarina contra Fusobacterium nucleatum. Se realizó extracción por expresión del aceite esencial de cáscaras de mandarina (variedades Arrayana y Oneco). Se evaluaron concentraciones al 20%, 40%, 60%, 80% y 100% del aceite esencial diluido en Tween al 0,02%. El efecto bacteriostático y bactericida se determinó por pruebas de sensibilidad antimicrobiana por difusión en disco. Como control positivo se utilizó Clorhexidina 0,2% y agua como control negativo. Se midió halo de inhibición (mm) y se determinó ausencia o presencia de crecimiento bacteriano a partir de unidades formadoras de colonias. Para comparación de proporciones de la actividad bacteriostática y bactericida, se realizó prueba de Fisher y T student (IC 95% p = 0,05). El halo de inhibición a una concentración del 100% mostró comportamiento similar a clorhexidina (p<0,05). Concentraciones al 100% y 80% fueron bactericidas, al 60%, 40% y 20% presentaron comportamiento bacteriostático. No se encontraron diferencias significativas en las proporciones de inhibición entre las dos variedades de mandarina (p>0,05). El uso de aceites esenciales de mandarina podría ser una alternativa complementaria al tratamiento de la enfermedad periodontal.
Abstract Chlorhexidine as a treatment of periodontal disease has achieved bactericidal effects over periodontopathogens and oral biofilm. Its use generates adverse effects; therefore natural alternatives are presented with a similar antimicrobial effect. Essential oils have proved effective in controlling dental plaque without the adverse effects of chlorhexidine. The aim of this study was to determine the bacteriostatic and bactericidal effect of essential oil of tangerine against Fusobacterium nucleatum. The extraction of the essential oil was performed by expression of tangerine peels (Arrayana and Oneco varieties). Concentrations at 20%, 40%, 60%, 80% and 100% of the essential oil diluted in 0,02% Tween were evaluated. The bacteriostatic and bactericidal effect was determined by antimicrobial susceptibility testing by disk diffusion. As a positive control 0,2% chlorhexidine and water as negative control were used. Inhibition zone (mm) was measured and presence or absence of bacterial growth was determined from colony forming units. To compare proportions of bacteriostatic and bactericidal activity, Fisher and T student test (95% CI p = 0,05) were performed. The 100% concentration zone of inhibition showed a similar behavior as chlorhexidine (p <0,05). 100% and 80% concentrations were bactericides, 60%, 40% and 20% showed bacteriostatic behavior. No significant differences between the proportions of inhibition of the two varieties of tangerine (p> 0,05). The use of essential oils of tangerine could be a complementary alternative to treatment of periodontal disease.
RESUMO
Genetic manipulation of the filamentous fungus Penicillium camemberti has been limited by a lack of suitable genetics tools for this fungus. In particular, there is no available homologous transformation system. In this study, the nitrate reductase (niaD) and orotidine-5'-monophosphate decarboxylase (pyrG) genes from Penicillium camemberti were characterized, and their suitability as metabolic molecular markers for transformation was evaluated. The genes were amplified using PCR-related techniques, and sequenced. The niaD gene is flanked by the nitrite reductase (niiA) gene in a divergent arrangement, being part of the putative nitrate assimilation cluster in P. camemberti. pyrG presents several polymorphisms compared with a previously sequenced pyrG gene from another P. camemberti strain, but almost all are silent mutations. Southern blot assays indicate that one copy of each gene is present in P. camemberti. Northern blot assays showed that the pyrG gene is expressed in minimal and rich media, and the niaD gene is expressed in nitrate, but not in reduced nitrogen sources. The functionality of the two genes as transformation markers was established by transforming A. nidulans pyrG- and niaD-deficient strains. Higher transformation efficiencies were obtained with a pyrG-containing plasmid. This is the first study yielding a molecular and functional characterization of P. camemberti genes that would be useful as molecular markers for transformation, opening the way for the future development of a non-antibiotic genetic transformation system for this fungus.