Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia.

Rhipicephalus microplus is recognized as a tick species highly prevalent in cattle, with a wide pantropical distribution that seems to continue spreading geographically. However, its role as a biological vector has been scarcely studied in the livestock context. In this study, a 16S rRNA next-generation sequencing analysis was used to determine bacterial diversity in salivary glands and gut of R. microplus from two contrasting livestock agroecosystems in Antioquia, Colombia. Both the culture-independent approach (CI) and the culture-dependent (CD) approach were complementarily adopted in this study. A total of 341 unique OTUs were assigned, the richness showed to be higher in the Northern than in the Middle Magdalena region, and a high diversity was found at the phylum and genus levels in the samples obtained. With the CI approach, Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria were the most common phylum of bacteria regardless of the organ, or geographic origin of the specimens analyzed. While the relative abundance of bacteria at a phylum level with the CD approach varied between analyzed samples, the data obtained suggest that a high diversity of species of bacteria occurs in R. microplus from both livestock agroecosystems. Bacterial genera such as Anaplasma, Coxiella, and Ehrlichia, recognized for their implications in tick-borne diseases, were also detected, together with endosymbionts such as Lysinibacillus, previously reported as a potential tool for biological control. This information is useful to deepen the knowledge about microbial diversity regarding the relations between endosymbionts and pathogens and could facilitate the future development of epidemiological surveillance in livestock systems.

Synthesis and characterization of a silver nanoparticle-containing polymer composite with antimicrobial abilities for application in prosthetic and orthotic devices.

BACKGROUND: The presence of skin problems in patients using external lower limb prosthesis is recurrent. This has generated the need to develop interfaces for prosthesis with the ability to control microbial growth. Silver nanoparticles (AgNPs) have been implemented in the development of biomaterials because of their high antimicrobial activity. This article discusses the development of an AgNP-containing polymer composite with antimicrobial activity for developing prosthetic liners. METHODS: AgNPs were synthesized using a photochemical method and certain physicochemical properties were characterized. Furthermore, the antimicrobial activity of AgNPs against Staphylococcus aureus ATCC 25923 and methicillin-resistant Staphylococcus aureus (MRSA), was assessed on the basis of their minimum inhibitory concentrations (MICs). AgNPs were incorporated into a silicon elastomer to assess certain physicomechanical properties, antimicrobial activity and cytotoxic effect of the material. RESULTS: The maximum antimicrobial activity of the material against Staphylococcus aureus ATCC 25923 and MRSA was 41.58% ±2.97% at AgNP concentration of 32.98 µg/mL and 14.85% ±5.94% at AgNP concentration of 16.49 µg/mL, respectively. Additionally, the material exhibited tensile yield strength, rupture tensile strength, and tensile modulus of elasticity of 0.70 - 1.10 MPa, 0.71-1.06 MPa, and 0.20 - 0.30 MPa, respectively. The mechanical characteristics of the material were within the acceptable range for use in external lower limb prosthetic and orthotic interfaces. CONCLUSIONS: It was possible to incorporate the AgNPs in a silicone elastomer, finding that the composite developed presented antimicrobial activity against Staphylococcus aureus ATCC 25923 and MRSA when compared to non-AgNP material samples.

Optimization of silver nanoparticle synthesis by chemical reduction and evaluation of its antimicrobial and toxic activity.

BACKGROUND: Chemical reduction has become an accessible and useful alternative to obtain silver nanoparticles (AgNPs). However, its toxicity capacity depends on multiple variables that generate differences in the ability to inhibit the growth of microorganisms. Thus, optimazing parameters for the synthesis of AgNPs can increase its antimicrobial capacity by improving its physical-chemical properties. METHODS: In this study a Face Centered Central Composite Design (FCCCD) was carried out with four parameters: A g N O 3 concentration, sodium citrate (TSC) concentration, N a B H 4 concentration and the pH of the reaction with the objective of inhibit the growth of microorganisms. The response variables were the average size of AgNPs, the peak with the greatest intensity in the size distribution, the polydispersity of the nanoparticle size and the yield of the process. AgNPs obtained from the optimization were characterized physically and chemically. The antimicrobial activity of optimized AgNPs was evaluated against Staphylococcus aureus, Escherichia coli, Escherichia coli AmpC resistant, and Candida albicans and compared with AgNPs before optimization. In addition, the cytotoxicity of the optimized AgNPs was evaluated by the colorimetric assay MTT (3- (4,5- Dimethylthiazol- 2- yl)- 2, 5 - Diphenyltetrazolium Bromide). RESULTS: It was found that the four factors studied were significant for the response variables, and a significant model (p < 0.05) was obtained for each variable. The optimal conditions were 8 for pH and 0.01 M, 0.0 6M, 0.01 M for the concentration of TSC, A g N O 3, and N a B H 4, respectively. Optimized AgNPs spherical and hemispherical were obtained, and 67.66% of it had a diameter less than 10.30 nm. A minimum bactericidal concentration (MBC) and minimum fungicidal Concentration (MFC) of optimized AgNPs was found against Staphylococcus aureus, Escherichia coli, Escherichia coli AmpC resistant, and Candida albicans at 19.89, 9.94, 9.94, 2.08 µg/mL, respectively. Furthermore, the lethal concentration 50 (L C 50) of optimized AgNPs was found on 19.11 µg/mL and 19.60 µg/mL to Vero and NiH3T3 cells, respectively. CONCLUSIONS: It was found that the factors studied were significant for the variable responses and the optimization process used was effective to improve the antimicrobial activity of the AgNPs.

Diabetes mellitus in patients with Alzheimer's disease: clinical description and correlation with the APOE genotype in a sample population from the province of Antioquia, Colombia.

INTRODUCTION: Alzheimer's disease is a multifactorial disease affecting approximately twenty million people worldwide. Numerous variables are associated with increased risk of developing this severe neurological disorder. Among the risk factors, diabetes mellitus, and the ε4 isoform of the APOE gene have been amply demonstrated as increasing the risk of developing this disease. OBJECTIVE: To determine if a correlation exists between APOE genotype, diabetes mellitus and Alzheimer's disease. MATERIALS AND METHODS: Clinical studies were carried out by surveying the clinical histories in a group of patients in the province of Antioquia, Colombia. Forty-three Alzheimer's patients were compared with 43 control subjects, paired by age and gender. Commercially available methods were used to determine whether the patients had diabetes, and restriction enzyme-based genotyping was used to determine the APOE genotypes. RESULTS: The most common non-neurological comorbidities were: arterial hypertension, acute myocardial infarction, chronic obstructive pulmonary disease and hypothyroidism. From the many variables investigated, two were conclusive: (1) the presence of Alzheimer's disease was higher in patients with diabetes mellitus, and (2) no correlation between late-onset sporadic Alzheimer's disease and APOE was found in the target population. CONCLUSIONS: To detect any association with the APOE genotype, a study involving much a larger population samples must be undertaken.

PURIFICACIÓN DEL ANTÍGENO 38 kDa DE Mycobacterium tuberculosis Y SU POTENCIAL USO EN DIAGNÓSTICO MEDIANTE INMUNOSENSORES PIEZOELÉCTRICOS / Mycobacterium tuberculosis 38 kDa Antigen Purification and Potential Diagnostic Use by Piezoelectric Immunosensors

Un paso crucial en el desarrollo de un inmunosensor piezoeléctrico para la detección de tuberculosis (TB), es la selección y obtención de los inmunoreactivos empleados en el inmunoensayo y la estrategia para la biofuncionalización del transductor. Diversos estudios han reportado el uso del antígeno proteico 38kDa (Ag38kDa) de Mycobacterium tuberculosis (Mtb) como un buen biomarcador de la enfermedad y el cumplimiento de las características físicas y bioquímicas para ser inmovilizado por monocapas autoensambladas (SAMs), en la superficie del electrodo de oro de cristales piezoeléctricos. Un inmunosensor piezoeléctrico desarrollado a partir de un antígeno nativo purificado de Mtb podría ser un método alternativo simple para la detección de Mtb con ventajas de rapidez y reusabilidad, contribuyendo al control y el tratamiento oportuno de la enfermedad. En este estudio se presenta el proceso de purificación del Ag38kDa a partir de proteínas de secreción filtradas de cultivo (CFP) de Mtb para ser usado como inmunoreactivo con potencial aplicación en la detección de Mtb con inmunosensores piezoeléctricos. Se obtuvieron cristales funcionalizados mediante la técnica modificada de monocapas autoensambladas (SAMs), con el antígeno nativo purificado y CFP. Las superficies biofuncionalizadas fueron caracterizadas cualitativamente con microscopía de fuerza atómica (AFM) para validar las condiciones de optimización del protocolo de inmovilización con antígenos de secreción de Mtb. Estos cristales modificados pueden ser acoplados a un sistema de caracterización de un inmunosensor piezoeléctrico para la detección de Mtb mediante un inmunoensayo competitivo directo.

The selection and procurance of the immunoreagents used in the immunoassay and biofunctionalisation transducer strategy, are a key in the piezoelectric immunosensor development for the detection of tuberculosis (TB). Many have reported the use of 38kDa protein antigen (Ag38kDa) from Mycobacterium tuberculosis (Mtb) such as good biomarker of TB disease and compliance with physical and biochemical characteristics to be immobilized by self-assembled monolayers (SAMs), in the gold electrode of piezoelectrics crystals surfaces. A piezoelectric immunosensor developed from purified native antigens of Mtb may be an alternative simple method for detection of Mtb with speed and reusable advantages, contributing to the control and early treatment of disease. In this paper, the purification process of Ag38kDa Mtb from secretory proteins filtered culture (CFP) from Mtb is presented as an immunoreactive with potential application in the detection of Mtb by piezoelectric immunosensors. Functionalized crystals were obtained by using the modified self-assembled monolayers (SAMs) technique, with purified native antigen and CFP. The functionalized surfaces were qualitatively characterized using atomic force microscopy (AFM) in order to validate the immobilization protocol optimal conditions for secretion antigens from Mtb. These modified crystals may be coupled to piezoelectric immunosensor characterization system for detecting of Mtb by a direct competition immunoassay.

Diabetes mellitus in patients withAlzheimer´s disease: clinical descriptionand correlation with the APOEgenotype in a sample population from the province of Antioquia, Colombia / Diabetes mellitus en pacientes con enfermedad de Alzheimer: descripción clínica y correlación con el genotipo APOE en una muestra de población del departamento de Antioquia, Colombia

Introduction. Alzheimer´s disease is a multifactorial disease affecting approximately twenty million people worldwide. Numerous variables are associated with increased risk of developing this severe neurological disorder. Among the risk factors, diabetes mellitus, and the ε4 isoform of the APOE gene have been amply demonstrated as increasing the risk of developing this disease. Objective. To determine if a correlation exists between APOE genotype, diabetes mellitus and Alzheimer´s disease. Materials and methods. Clinical studies were carried out by surveying the clinical histories in a group of patients in the province of Antioquia, Colombia. Forty-three Alzheimer´s patients were compared with 43 control subjects, paired by age and gender. Commercially available methods were used to determine whether the patients had diabetes, and restriction enzyme-based genotyping was used to determine the APOE genotypes. Results. The most common non-neurological comorbidities were: arterial hypertension, acute myocardial infarction, chronic obstructive pulmonary disease and hypothyroidism. From the many variables investigated, two were conclusive: (1) the presence of Alzheimer´s disease was higher in patients with diabetes mellitus, and (2) no correlation between late-onset sporadic Alzheimer´s disease and APOE was found in the target population. Conclusions. To detect any association with the APOE genotype, a study involving much a larger population samples must be undertaken.

Introducción. La enfermedad de Alzheimer es compleja y afecta, aproximadamente, a 20 millones de personas en todo el mundo. Muchas variables parecen aumentar el riesgo de desarrollar esta alteración neurológica. Entre los factores de riesgo, se ha demostrado ampliamente que la diabetes mellitus y la isoforma ε4 del gen APOE tienen incidencia positiva en el desarrollo de la enfermedad. Se reporta un estudio en el cual se investigó la posible correlación entre APOE, diabetes mellitus y la enfermedad de Alzheimer, en un grupo específico de pacientes del departamento de Antioquia, Colombia. Objetivo. Determinar si existe una correlación entre APOE, diabetes mellitus y la enfermedad de Alzheimer, en un grupo de pacientes de Antioquia, Colombia. Materiales y métodos. Se buscaron y analizaron las historias clínicas de los pacientes con diagnóstico de enfermedad de Alzheimer. Se seleccionaron aquellos que cumplían los criterios de inclusión. Se utilizaron métodos comercialmente disponibles para confirmar la presencia de diabetes mellitus. La genotipificación de APOE se hizo con un método basado en la PCR y la digestión con enzimas de restricción, en muestras de todos los participantes en el estudio. Resultados. En este estudio se analizan 43 casos de enfermedad de Alzheimer y 43 individuos sanos controles, pareados por edad y sexo. Las enfermedades concomitantes no neurológicas más comunes fueron: hipertensión arterial, infarto agudo del miocardio, enfermedad pulmonar obstructiva crónica e hipotiroidismo. Conclusiones. De las diferentes variables investigadas, dos arrojaron resultados concluyentes: i) la presencia de la enfermedad de Alzheimer es más frecuente en pacientes con diabetes mellitus, y 2) no se encontró correlación entre la enfermedad de Alzheimer de inicio tardío esporádico y el genotipo de APOE. Es importante indicar que debe llevarse a cabo un estudio con un tamaño de población mayor, para determinar cualquier posible correlación o inferencia con el genotipo de APOE.