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

Cytotoxicity of a Lipid-Rich Extract from Native Mexican Avocado Seed (Persea americana var. drymifolia) on Canine Osteosarcoma D-17 Cells and Synergistic Activity with Cytostatic Drugs

Osteosarcoma is the most common malignant bone tumor in both children and dogs. It is an aggressive and metastatic cancer with a poor prognosis for long-term survival. The search for new anti-cancer drugs with fewer side effects has become an essential goal for cancer chemotherapy; in this sense, the bioactive compounds from avocado have proved their efficacy as cytotoxic molecules. The objective of this study was to determine the cytotoxic and antiproliferative effect of a lipid-rich extract (LEAS) from Mexican native avocado seed (Persea americana var. drymifolia) on canine osteosarcoma D-17 cell line. Also, the combined activity with cytostatic drugs was evaluated. LEAS was cytotoxic to D-17 cells in a concentration-dependent manner with an IC50 = 15.5 µg/mL. Besides, LEAS induced caspase-dependent cell apoptosis by the extrinsic and intrinsic pathways. Moreover, LEAS induced a significant loss of mitochondrial membrane potential and increased superoxide anion production and mitochondrial ROS. Also, LEAS induced the arrest of the cell cycle in the G0/G1 phase. Finally, LEAS improved the cytotoxic activity of cisplatin, carboplatin, and in less extension, doxorubicin against the canine osteosarcoma cell line through a synergistic effect. In conclusion, avocado could be a potential source of bioactive molecules in the searching treatments for osteosarcoma

Molecular characterization of bacterial microbiota associated with infectious bovine keratoconjunctivitis in Michoacán, Mexico

Background: The most common ocular disease affecting cattle worldwide is infectious bovine keratoconjunctivitis (IBK), which has been associated with Moraxella bovis bacterium. Objective: To report the molecular characterization of the ocular bacterial microbiota and its relation to IBK in cattle in two dairy regions in Michoacán, Mexico. Methods: A total population of 761 bovines were evaluated, of which 17 (2.23%) showed symptoms of IBK. Thirty-eight bacterial isolates from ocular samples of bovines with IBK were characterized by Gram-staining and antimicrobial sensitivity. In addition, isolates were identified by sequence comparisons of the 16S ribosomal gene. Results: The genus Moraxella was one of the most abundant bacteria and M. bovoculi was the most predominant species. Conclusion: The bacterial isolates identified in eye lesions of cattle and associated to IBK are diverse. To the author ́s knowledge, this is the first study on the subject in Mexico; therefore, more research is needed to estimate the incidence of IBK and determine its associated microbiota.Antecedentes: a doença ocular mais comum que afeta o gado no mundo é a ceratoconjuntivite bovina (IBK), que tem sido associada à bactéria Moraxella bovis. Objetivo: relatar a caracterização molecular da microbiota bacteriana ocular e sua relação com a IBK em bovinos de duas regiões leiteiras de Michoacán, México. Métodos: foi avaliada uma população total de 761 bovinos, mas apenas 17 (2,23%) apresentaram sintomas de IBK. Trinta e oito isolados bacterianos de amostras de olho bovino com IBK foram caracterizados por coloração de Gram e sensibilidade antimicrobiana. Além disso, os isolados foram identificados por comparação de sequências do gene ribossômico 16S. Resultados: a microbiota bacteriana associada à IBK foi diversa, sendo o gênero Moraxella uma das mais abundantes e M. bovoculi a espécie predominante. Conclusão: de acordo com o conhecimento dos autores, este é o primeiro estudo sobre o tema no México até o momento, portanto é necessário expandir essa pesquisa para estimar a incidência de IBK e determinar a microbiota associada à mesma.Antecedentes: la enfermedad ocular más común que afecta al ganado en todo el mundo es la queratoconjuntivitis infecciosa bovina (IBK), que se ha asociado con la bacteria Moraxella bovis. Objetivo: reportar la caracterización molecular de la microbiota bacteriana ocular y su relación con IBK en ganado de dos regiones lecheras en Michoacán, México. Métodos: se evaluó una población total de 761 bovinos de los cuales 17 (2,23%) mostraron síntomas de IBK. Se obtuvieron treinta y ocho aislamientos bacterianos de muestras oculares de bovinos con IBK, los cuales se caracterizaron por tinción de Gram y sensibilidad antimicrobiana. Además, los aislamientos se identificaron mediante comparaciones de secuencias del gen ribosomal 16S. Resultados: el género Moraxella fue una de las bacterias más abundantes y M. bovoculi fue la especie más predominante. Conclusión: los aislamientos bacterianos identificados en lesiones oculares de bovinos y asociados a IBK son diversos. Hasta donde sabemos, este es el primer estudio sobre el tema realizado en México; por lo tanto, es necesario ampliar esta investigación para estimar la incidencia de IBK y determinar la microbiota asociada con la misma

Compósitos de quitosano-ácidos grasos reducen la infección de Botrytis cinerea en fresa en poscosecha

Grey mold is one of the main causes of post-harvest strawberry (Fragaria x ananassa Duch.) rot fruit, which is caused by Botrytis cinerea Pers.:fr fungus. The treatment with chemical fungicides is the main method for its control; however, its use cause health concerns and pollution. The above promote the research toward alternatives of control. The aim of the present research was to evaluate the chitosan-fatty acids composites effects on the B. cinerea infection in post-harvest strawberry fruits. Method In vitro bioassays were conducted to evaluate the chitosan, sodium octanoate and decanoic acid effect on the mycelial growth of B. cinerea. Likewise, in vivo bioassays were conducted to evaluate the compounds alone and in mixture effect on infection severity of grey mold in post-harvest strawberry fruits. The treatments were evaluated in a completely randomized design and the data were analyzed by means of analysis of variance (ANOVA) (p ≤ 0.05) and average comparison by Tukey´s test (p ≤ 0.05). Results The chitosan inhibited 100 % of the growth when it was applied in 10, 12.5 and 15 mg/mL concentrations, while the sodium octanoate reached the same inhibition percentage in 0.33 and 0.49 mg/mL concentrations. Finally, decanoic acid inhibited in 100 % to B. cinerea in 0.17, 0.34 and 0.51 mg/mL concentrations. In in vivo bioassays, the best results of strawberry fruits protection from B. cinerea infection were those in which the chitosan/sodium octanoate (12.5/0.49 y 15/0.49 mg/mL) were utilized, with absence of severity. Also highlight the chitosan (15 mg/mL), decanoic acid (0.51 mg/mL) and chitosan/decanoic acid composites (12.5/0.51 and 15/0.51 mg/mL) treatments, with severity degrees between 1 and 2. Discussion or Conclusion The chitosan, sodium octanoate and decanoic acid significantly inhibited the in vitro mycelial growth of B. cinerea. The chitosan/sodium octanoate composite have a greater protective effect of strawberry fruits, in relation to the compounds applied alone. The chitosan and decanoic acid presented a significantly protective effect of the fruits, applied alone as well as in composite. The above suggest that these compounds could be potentially used in the post-harvest control of B. cinerea.El moho gris es una de las principales causas de pudrición en poscosecha del fruto de fresa (Fragaria x ananassa Duch.), la cual es ocasionada por el hongo Botrytis cinerea Pers.:fr. El tratamiento con fungicidas químicos es el método principal de su control; no obstante, su uso ocasiona problemas de salud y contaminación. Lo anterior promueve la investigación hacia alternativas de control. El objetivo de la presente investigación fue evaluar el efecto de compósitos de quitosano-ácidos grasos sobre la infección de B. cinerea en frutos de fresa en poscosecha. Método Se realizaron bioensayos in vitro para evaluar el efecto del quitosano, octanoato de sodio y ácido decanoico sobre el crecimiento micelial de B. cinerea. Asimismo, en bioensayos in vivo se evaluó la protección de los compuestos solos y en mezcla sobre la severidad de la infección del moho gris en frutos de fresa en poscosecha. Los tratamientos se evaluaron bajo un diseño completamente al azar, y los datos se analizaron mediante análisis de varianza (ANOVA) (p ≤ 0.05) y comparación de medias por el método de Tukey (p ≤ 0.05). Resultados El quitosano inhibió el crecimiento en 100 % cuando se aplicó en concentraciones de 10, 12.5 y 15 mg/mL, mientras que el octanoato de sodio alcanzó el mismo porcentaje de inhibición en las concentraciones de 0.33 y 0.49 mg/mL. Finalmente, el ácido decanoico inhibió en 100 % a B. cinerea en las concentraciones de 0.17, 0.34 y 0.51 mg/mL. En los bioensayos in vivo, los mejores resultados de protección de los frutos de fresa de la infección por B. cinerea fueron aquellos en los que se utilizó el compósito quitosano/octanoato de sodio (12.5/0.49 y 15/0.49 mg/mL) con ausencia de severidad. También destacaron los tratamientos de quitosano (15 mg/mL), ácido decanoico (0.51 mg/mL) y del compósito quitosano/ácido decanoico (12.5/0.51 y 15/0.51 mg/mL), donde el grado de severidad fue entre 1 y 2. Discusión o Conclusión El quitosano, octanoato de sodio y ácido decanoico inhibieron significativamente el crecimiento micelial in vitro de B. cinerea. El compósito quitosano/octanoato de sodio posee un efecto protector mayor de los frutos de fresa, en relación a los compuestos aplicados solos. El quitosano y el ácido decanoico presentaron un efecto protector significativo de los frutos, aplicados solos o como compósito. Lo anterior sugiere que estos compuestos podrían ser utilizados potencialmente en el control de B. cinerea en poscosecha

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

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

Expression analysis of the Arabidopsis thaliana AtSpen2 gene, and its relationship with other plant genes encoding Spen proteins

"Proteins of the Split ends (Spen) family are characterized by an N-terminal domain, with one or more RNA recognition motifs and a SPOC domain. In Arabidopsis thaliana, the Spen protein FPA is involved in the control of flowering time as a component of an autonomous pathway independent of photoperiod. The A. thaliana genome encodes another gene for a putative Spen protein at the locus At4g12640, herein named AtSpen2. Bioinformatics analysis of the AtSPEN2 SPOC domain revealed low sequence similarity with the FPA SPOC domain, which was markedly lower than that found in other Spen proteins from unrelated plant species. To provide experimental information about the function of AtSpen2, A. thaliana plants were transformed with gene constructs of its promoter region with uidA::gfp reporter genes; the expression was observed in vascular tissues of leaves and roots, as well as in ovules and developing embryos. There was absence of a notable phenotype in knockout and overexpressing lines, suggesting that its function in plants might be specific to certain endogenous or environmental conditions. Our results suggest that the function of Atspen2 diverged from that of fpa due in part to their different transcription expression pattern and divergence of the regulatory SPOC domain.

Jet-hadron correlations measured relative to the second order event plane in Pb–Pb collisions at √sNN = 2.76 TeV

The Quark Gluon Plasma (QGP) produced in ultra relativistic heavy-ion collisions at the Large Hadron Collider (LHC) can be studied by measuring the modifications of jets formed by hard scattered partons which interact with the medium. We studied these modifications via angular correlations of jets with charged hadrons for jets with momenta 20 < pjetT < 40 GeV/c as a function of the associated particle momentum. The reaction plane fit (RPF) method is used in this analysis to remove the flow modulated background. The analysis of angular correlations for different orientations of the jet relative to the second order event plane allows for the study of the path length dependence of medium modifications to jets. We present the dependence of azimuthal angular correlations of charged hadrons with respect to the angle of the axis of a reconstructed jet relative to the event plane in Pb-Pb collisions at sNN−−−√ = 2.76 TeV. The dependence of particle yields associated with jets on the angle of the jet with respect to the event plane is presented. Correlations at different angles relative to the event plane are compared through ratios and differences of the yield. No dependence of the results on the angle of the jet with respect to the event plane is observed within uncertainties, which is consistent with no significant path length dependence of the medium modifications for this observable