Evaluación in vitro de múltiples agentes antibacterianos para el tratamiento de la blefaritis anterior crónica estafilocócica / In vitro evaluation of multiple antibacterial agents for the treatment of chronic staphylococcal anterior blepharitis

Objetivo Evaluar la eficacia bactericida de varios compuestos utilizados en el tratamiento de la blefaritis anterior estafilocócica crónica mediante un estudio in vitro. Materiales y métodos Se cultivaron cepas comerciales estándar de Staphylococcus aureus (SAu) (ATCC 25923 Culti-Loops) y Staphylococcus coagulasa-negativo (CoNS) (ATCC 12228 Culti-Loops). Se realizaron pruebas de sensibilidad a vancomicina 30μg, netilmicina 30μg, ácido hipocloroso (HOCl) al 0,01% (Ocudox™, Brill®), aceite de hoja de Melaleuca alternifolia (MeAl) (Navyblef® Cuidado diario, NOVAX®) y digluconato de clorhexidina al 1% (DGCH) (Cristalmina™, Salvat®) mediante el método de difusión en disco de agar (Rosco Neo-Sensitabs™). A las 24horas se midieron los halos inducidos con calibradores automáticos. Los resultados se analizaron con las guías EUCAST- y CLSI potency Neo-Sensitabs™. Resultados La vancomicina indujo un halo de 22,37mm y 21,81mm en SAu y CoNS, respectivamente. La netilmicina produjo halos de 24,45mm en SAu y de 32,49mm en CoNS. MeAl indujo halos de 12,65mm en SAu y de 15,83mm en CoNS. Se encontró un halo de 12,11mm en SAu y un halo de 18,38mm en CoNS utilizando HOCl. DGCH produjo halos de 26,55mm y 23,12mm en SAu y CoNS, respectivamente. Conclusión La netilmicina y la vancomicina demostraron actividad antibiótica frente a ambos patógenos, por lo que pueden ser terapias alternativas de rescate para tratar la blefaritis estafilocócica crónica. El DGCH presenta una eficacia frente a ambos comparable a los antibióticos, mientras que el HOCl y la MeAl demuestran menor eficacia (AU)

Objective To evaluate the bactericidal efficacy of several compounds used in the treatment of chronic staphylococcal anterior blepharitis through an in vitro study. Materials and methods Standard commercial strains of Staphylococcus aureus (SAu) (ATCC 25923 Culti-Loops) and coagulase-negative Staphylococcus (CoNS) (ATCC 12228 Culti-Loops) were cultured. Susceptibility tests were performed to vancomycin 30μg, netilmicin 30μg, hypochlorous acid (HOCl) 0.01% (Ocudox™, Brill®), Melaleuca alternifolia leaf oil (MeAl) (Navyblef® Daily Care, NOVAX®) and 1% chlorhexidine digluconate (DGCH) (Cristalmina™, Salvat®) using the agar disk diffusion method (Rosco Neo-Sensitabs®). After 24hours, the induced halos were measured with automatic calipers. The results were analyzed using the EUCAST- and CLSI potency Neo-Sensitabs® guidelines. Results Vancomycin induced a halo of 22.37mm and 21.81mm in SAu and CoNS, respectively. Netilmicin produced halos of 24.45mm in SAu and 32.49mm in CoNS. MeAl induced halos of 12.65mm in SAu and 15.83mm in CoNS. A 12.11mm halo was found in SAu and an 18.38mm halo in CoNS using HOCl. DGCH produced halos of 26.55mm and 23.12mm in SAu and CoNS, respectively. Conclusion Netilmicin and vancomycin demonstrated antibiotic activity against both pathogens, so they can be alternative rescue therapies to treat chronic staphylococcal blepharitis. DGCH has efficacy against both comparable to antibiotics, while HOCl and MeAl show less efficacy (AU)

In vitro evaluation of multiple antibacterial agents for the treatment of chronic staphylococcal anterior blepharitis.

OBJECTIVE: To evaluate the bactericidal efficacy of several compounds used in the treatment of chronic staphylococcal anterior blepharitis through an in vitro study. MATERIALS AND METHODS: Standard commercial strains of Staphylococcus aureus (SAu) (ATCC 25923 Culti-Loops) and coagulase-negative Staphylococcus (CoNS) (ATCC 12228 Culti-Loops) were cultured. Susceptibility tests were performed to vancomycin 30 µg, netilmicin 30 µg, hypochlorous acid (HOCl) 0.01% (Ocudox™, Brill®), Melaleuca alternifolia leaf oil (MeAl) (Navyblef® Daily Care, NOVAX®) and 1% chlorhexidine digluconate (DGCH) (Cristalmina™, Salvat®) using the agar disk diffusion method (Rosco Neo-Sensitabs®). After 24 h, the induced halos were measured with automatic calipers. The results were analyzed using the EUCAST- and CLSI potency Neo-Sensitabs® guidelines. RESULTS: Vancomycin induced a halo of 22.37 mm and 21.81 mm in SAu and CoNS, respectively. Netilmicin produced halos of 24.45 mm in SAu and 32.49 mm in CoNS. MeAl induced halos of 12.65 mm in SAu and 15.83 mm in CoNS. A 12.11 mm halo was found in SAu and an 18.38 mm halo in CoNS using HOCl. DGCH produced halos of 26.55 mm and 23.12 mm in SAu and CoNS, respectively. CONCLUSION: Netilmicin and vancomycin demonstrated antibiotic activity against both pathogens, so they can be alternative rescue therapies to treat chronic staphylococcal blepharitis. DGCH has efficacy against both comparable to antibiotics, while HOCl and MeAl show less efficacy.

Formación de biopelículas por micoplasmas de importancia médica / Biofilm formation by mycoplasmas of medical importance

El objetivo de este estudio fue evaluar la formación de biopelículas por micoplasmas de interés médico.Métodos. La formación de las biopelículas se realizó en microplacas, fueron enjuagadas con solución PBS para remover las células que no se adhirieron y se tiñeron con soluciónde cristal violeta al 0,5% durante 30 minutos. La formación de biopelículas por parte de Mycoplasma pneumoniae, Mycoplasma fermentans y Mycoplasma penetrans se analizó por medio de microscopía óptica y microscopía electrónica de barrido. Resultados. Los micoplasmas evaluados mostraron la capacidad para formar biopelículas,lo cual se evidenció por medio de la tinción de cristal violeta. Las biopelículas formadas en las microplacas y analizadas por microscopía mostraron agregados de microcolonias. La formación de biopelículas por parte de M. fermentans, M. pneumoniae y M. penetrans se presentó a las 72 horas de incubación. Se comparó la formación de biopelícula y la cuantificación de plancton, y se encontró correlación. Conclusión. Se debe considerar que este estudio se hizo bajo condiciones de laboratorioy que, para trabajos futuros, se recomienda utilizar modelos animales para definir cómo contribuyen estos agregados en la persistencia en los huéspedes. Estos resultados sugierenque la capacidad de M. fermentans, M. pneumoniae y M. penetrans para formar biopelículas puede considerarse un factor de virulencia, y un evento importante en la patogénesis yevolución en infecciones asociadas con el uso de dispositivos médicos.

The purpose of this study was to evaluate the development of biofilms by mycoplasmas of medical importance.Methods: Biofilms grown in microtiter plates were rinsed briefly in PBS to remove non-adherent cells and stained with 0,5% crystal violet solution for 30 minutes. The biofilm formation bymycoplasma species were analyzed by optical and scanning electron microscopy. Results: Three mycoplasma species were assessed for their ability to form biofilms. Crystal violet staining of biofilms in microtiter plates revealed the ability of mycoplasma to form a biofilm. Microscopic analysis of crystalviolet stained biofilms on microtiters indicated aggregation to form microcolonies. Biofilm growth by Mycoplasma fermentans, Mycoplasmapneumoniae and Mycoplasmapenetrans was followed over a time course of 72 hours. Mycoplasma were also analyzed quantitatively for biofilm formation and cell counts compared for both biofilm and plankton cells. Cell counts for biofilms showed a goodcorrelation with results obtained using crystal violet staining. Conclusion: This study has examined biofilm formation under in vitro laboratory conditions.Further studies on animal models will be crucial to determine if biofilms form in vivo and whether they contribute to mycoplasma persistence in thehost. These results suggest that the ability of M. fermentans, M. pneumoniae and M. penetrans to form a biofilm may be a virulence factor, and an important event in the pathogenesis and evolutionin infections associated with the use of medical devices.