Antimicrobial effects of plant defence peptides expressed by bovine endothelial cells on intracellular pathogens

Background: The actions of plant antimicrobial peptides (PAP) against intracellular pathogens are poorly known. It has been reported that wheat puroindolines show antibacterial activity against Staphylococcus epidermidis endocyted by macrophages. In this work, we evaluated the intracellular antimicrobial activity of PAP \u3b3-thionin and thionin Thi2.1 produced by bovine endothelial cells against intracellular Staphylococcus aureus and Candida albicans . We used three host-pathogen models: 1) bovine mammary epithelial cells (BMEC)-S. aureus, 2) bovine endothelial cells (BEC)-S. aureus and 3) BEC-C. albicans, and evaluated the effect of conditioned media from BEC producers of PAP (\u3b3-thionin and thionin Thi2.1). Results: In the first model, conditioned medium (CM) containing Thi2.1 completely inhibited S. aureus intracellular after 24 hrs treatment. In the second model, CM from BEC containing \u3b3-thionin has a better effect killing intracellular S. aureus for 12-24 hrs incubations than CM from endothelial cells producers of Thi2.1; this was related with an increase of nitric oxide production (~2 times) in BEC infected and treated for 12 hrs with CM containing \u3b3-thionin, which negatively correlates with bacterial viability. In the third model, CM containing Thi2.1 showed a more potent intracellular fungicidal activity (~85% of inhibition) at 24 hrs treatment than CM containing \u3b3-thionin (~35% of inhibition). Conclusions: This work shows new effects of PAP to control intracellular bacterial or fungal infections

Flora etnomedicinal utilizada para el tratamiento de afecciones dermatológicas en la Meseta Purépecha, Michoacán, México

Background and Aims: Inhabitants of the Purépecha Plateau preserve an excellent ancestral knowledge on medicinal plants used for dermatological affections, which has not been documented. An ethnopharmacological survey was carried out in this region to gather information on the use of medicinal plants and herbal preparations for treating dermatological affections, to disseminate the Purépecha indigenous knowledge and identifying promising plants for developing new formulations for cutaneous conditions.Methods: The study was conducted in the 21 municipalities that compose the Purépecha Plateau. A total of 86 local inhabitants (62 women and 24 men) were interviewed. The data were quantitatively analyzed through the determination of the use value, fidelity level and informant consensus factor.Key results: A total of 97 plant species belonging to 47 families were documented for treating 19 dermatological conditions on the Purépecha Plateau. Asteraceae was the leading family among the collected medicinal plants (20.61%), followed by Lamiaceae (13.40%) and Solanaceae (5.15%). The largest number of plants was used for the treatment of cuts (40.20%), skin inflammation (37.11%) and rash (37.11%). The aerial parts were the most commonly used (34.75%). The medicinal plant species with larger use values were Heterotheca inuloides (0.53), Aloe vera (0.37) and Oenothera rosea (0.21). The comparison of results with ethnomedicinal literature worldwide revealed that 8.25% of plants used on the Purépecha Plateau were recorded for the first time for the treatment of dermatological affections.Conclusions: This study provides new information on medicinal plants used on the Purépecha Plateau to treat cutaneous diseases. Future pharmacological and toxicological investigations are required to demonstrate the efficacy and safety of these species for treating dermatological affections.Antecedentes y Objetivos: Los habitantes de la Meseta Purépecha conservan un excelente conocimiento ancestral sobre plantas medicinales utilizadas para tratar afecciones dermatológicas que no ha sido documentado. Un estudio etnofarmacológico fue realizado en esta región para recopilar información sobre el uso de plantas medicinales y preparaciones herbarias utilizadas en la zona para el tratamiento de afecciones dermatológicas, con el objetivo de difundir la medicina tradicional Purépecha e identificar plantas prometedoras en el desarrollo de tratamientos para afecciones cutáneas.Métodos: El estudio se realizó en los 21 municipios que componen la Meseta Purépecha. Se entrevistó a un total de 86 habitantes locales (62 mujeres y 24 hombres). Los datos se analizaron cuantitativamente mediante la determinación del valor de uso, nivel de fidelidad y el factor de consenso informante.Resultados clave: En total 97 especies de plantas pertenecientes a 47 familias fueron documentadas para el tratamiento de 19 afecciones dermatológicas en la Meseta Purépecha. Asteraceae fue la familia principal entre las plantas colectadas (20.61%), seguida de Lamiaceae (13.40%) y Solanaceae (5.15%). La mayor cantidad de plantas se utilizó para tratamiento de heridas (40.20%), inflamación de la piel (37.11%) y erupciones cutáneas (37.11%). La principal parte utilizada de la planta fue la aérea (34.75%). Las plantas medicinales con mayores valores de uso fueron Heterotheca inuloides (0.53), Aloe vera (0.37) y Oenothera rosea (0.21). La comparación de los resultados con la literatura etnomedicinal reveló que 8.25% de las plantas utilizadas en la Meseta Purépecha se registraron por primera vez para tratamiento de afecciones dermatológicas.Conclusiones: Este estudio proporciona nueva información sobre plantas medicinales utilizadas en la Meseta Purépecha para tratamiento de enfermedades cutáneas. Se requieren futuras investigaciones farmacológicas y toxicológicas para demostrar la eficacia y seguridad de estas especies para el tratamiento de afecciones dermatológicas

Caracterización molecular de aislamientos de Staphylococcus spp. asociados a mastitis bovina en Tarímbaro, Michoacán

Mastitis is a disease of dairy cattle worldwide that causes decreased milk quality/yield, resulting in major economic losses. Mastitis is caused by a wide variety of organisms, including Staphylococcus aureus as a major pathogen. The purpose of this research was to characterize 20 Staphylococcus spp. isolates from the mammary gland of mastitic, lactating, backyard cattle, using both the Random Amplified Polymorphic DNA (RAPD) analysis, and sensitivity tests to 13 antibiotics and 1 detergent. Sensitivity tests to 13 antibiotics showed that all isolates were resistant to penicillin but sensitive to cephalothin, erythromycin, and trimethoprim. All isolates were susceptible to a 90% similarity in the RAPD band pattern, while isolates SA35 & SA36 were identical to each other. Nevertheless, all other isolates showed different amplification profiles. Isolates SA7 & SA8, SA30 & SA32, SA35 & SA36 had similar antibiotic resistance/RAPD patterns, suggesting re-isolation. Staphylococcus isolates collected in Tarímbaro, Michoacán showed several different amplification patterns.La mastitis es una enfermedad que afecta al ganado bovino lechero a nivel mundial, provocando una disminución en la calidad y cantidad de la leche producida, lo cual genera pérdidas económicas considerables. Esta enfermedad es causada por una variedad de microorganismos, entre los cuales Staphyloccocus aureus es uno de los más importantes. El objetivo del trabajo fue caracterizar 20 aislamientos de Staphylococcus spp. obtenidos de la glándula mamaria de vacas lactantes con mastitis en un sistema de producción de traspatio, mediante el uso de polimorfismos derivados de la amplificación al azar de ADN (RAPD's) y su sensibilidad hacia trece antibióticos y 1 detergente. Las pruebas de sensibilidad a los 13 antibióticos demostraron que todos los aislamientos fueron resistentes a penicilina y sensibles a cefalotina, eritromicina y trimetoprim. Todos los aislamientos fueron susceptibles a una concentración menor de 6 µg/ml de bromuro de cetil trimetil amonio (CTAB). La caracterización molecular permitió determinar que los aislamientos SA7 y SA8, SA30 y SA32, presentaron una similitud superior al 90 % en el patrón de bandas obtenida por RAPD, mientras que los aislamientos SA35 y SA36 fueron idénticos, sin embargo, el resto de los aislamientos presentaron perfiles de amplificación distintos. Los aislamientos SA7 y SA8, SA30 y SA32, SA35 y SA36, tuvieron patrones de resistencia a antibióticos y de RAPD similares, por lo que probablemente se trató de reaislamientos. Los aislamientos de Staphylococcus de Tarí­mbaro, Michoacán presentaron una diversidad de perfiles de amplificación

Nonprofessional Phagocytic Cell Receptors Involved in Staphylococcus aureus Internalization

Staphylococcus aureus is a successful human and animal pathogen. The majority of infections caused by this pathogen are life threatening, primarily because S. aureus has developed multiple evasion strategies, possesses intracellular persistence for long periods, and targets the skin and soft tissues. Therefore, it is very important to understand the mechanisms employed by S. aureus to colonize and proliferate in these cells. The aim of this review is to describe the recent discoveries concerning the host receptors of nonprofessional phagocytes involved in S. aureus internalization. Most of the knowledge related to the interaction of S. aureus with its host cells has been described in professional phagocytic cells such as macrophages. Here, we showed that in nonprofessional phagocytes the α5β1 integrin host receptor, chaperons, and the scavenger receptor CD36 are the main receptors employed during S. aureus internalization. The characterization and identification of new bacterial effectors and the host cell receptors involved will undoubtedly lead to new discoveries with beneficial purposes

Effects of Sodium Octanoate on Innate Immune Response of Mammary Epithelial Cells during Staphylococcus aureus Internalization

Bovine mammary epithelial cells (bMECs) are capable of initiating an innate immune response to invading bacteria. Short chain fatty acids can reduce Staphylococcus aureus internalization into bMEC, but it has not been evaluated if octanoic acid (sodium octanoate, NaO), a medium chain fatty acid (MCFA), has similar effects. In this study we determined the effect of NaO on S. aureus internalization into bMEC and on the modulation of innate immune elements. NaO (0.25–2 mM) did not affect S. aureus growth and bMEC viability, but it differentially modulated bacterial internalization into bMEC, which was induced at 0.25–0.5 mM (~60%) but inhibited at 1-2 mM (~40%). Also, bMEC showed a basal expression of all the innate immune genes evaluated, which were induced by S. aureus. NaO induced BNBD4, LAP, and BNBD10 mRNA expression, but BNBD5 and TNF-α were inhibited. Additionally, the pretreatment of bMEC with NaO inhibited the mRNA expression induction generated by bacteria which coincides with the increase in internalization; only TAP and BNDB10 showed an increase in their expression; it coincides with the greatest effect on the reduction of bacterial internalization. In conclusion, NaO exerts a dual effect on S. aureus internalization in bMEC and modulates elements of innate immune response

Antimicrobial effects of plant defence peptides expressed by bovine endothelial cells on intracellular pathogens

Background: The actions of plant antimicrobial peptides (PAP) against intracellular pathogens are poorly known. It has been reported that wheat puroindolines show antibacterial activity against Staphylococcus epidermidis endocyted by macrophages. In this work, we evaluated the intracellular antimicrobial activity of PAP γ-thionin and thionin Thi2.1 produced by bovine endothelial cells against intracellular Staphylococcus aureus and Candida albicans . We used three host-pathogen models: 1) bovine mammary epithelial cells (BMEC)-S. aureus, 2) bovine endothelial cells (BEC)-S. aureus and 3) BEC-C. albicans, and evaluated the effect of conditioned media from BEC producers of PAP (γ-thionin and thionin Thi2.1). Results: In the first model, conditioned medium (CM) containing Thi2.1 completely inhibited S. aureus intracellular after 24 hrs treatment. In the second model, CM from BEC containing γ-thionin has a better effect killing intracellular S. aureus for 12-24 hrs incubations than CM from endothelial cells producers of Thi2.1; this was related with an increase of nitric oxide production (~2 times) in BEC infected and treated for 12 hrs with CM containing γ-thionin, which negatively correlates with bacterial viability. In the third model, CM containing Thi2.1 showed a more potent intracellular fungicidal activity (~85% of inhibition) at 24 hrs treatment than CM containing γ-thionin (~35% of inhibition). Conclusions: This work shows new effects of PAP to control intracellular bacterial or fungal infections