RESUMEN
Abstract Acidipropionibacterium acidipropionici is widely used for many applications, suchas propionic acid production, cereal silage, and also as probiotic. Due to this plethora of appli-cations, new isolates of A. acidipropionici with improved features are being searched for. Thesenew isolates must be accurately identified, however, most approaches become expensive andtime-consuming when the number of isolates is high. On the contrary, fluorescence in situhybridization allows the affordable, reliable, and rapid identification of microorganisms in purecultures and environmental and medical samples. Therefore, the aim of this work was to applya fluorescent in situ hybridization probe for the reliable identification of new A. acidipropioniciisolates. To this end, probe Pap446, specific for A. acidipropionici, was validated by hybridiza-tion assays with strains of this species from different origins, other species of the same genusor family, and unrelated genera. Eight isolates with propionibacterium characteristics wereobtained from milk and feces of cows. Probe Pap446, hybridized only with isolates III and VI.The identity of these isolates was further confirmed by PCR using group and species-specificprimers for propionibacteria and 16S rDNA sequencing.
Resumen Acidipropionibacterium acidipropionici es ampliamente usada para diversas aplicaciones, como producción de ácido propiónico, ensilado de cereales y probiótico. Debido a esta variedad de aplicaciones, continuamente se buscan nuevos aislamientos de A. acidipropionici con características nuevas. Estos nuevos aislamientos deben ser identificados correctamente, pero la mayoría de las técnicas disponibles resultan costosas e insumen mucho tiempo cuando el número de aislamientos es elevado. Por el contrario, la hibridación fluorescente in situ permite una identificación barata, confiable y rápida de microorganismos en cultivos puros y en muestras ambientales y médicas. Por lo tanto, el objetivo de este trabajo fue la aplicación de una sonda oligonucleotídica en un protocolo de hibridación fluorescente in situ para la identificación confiable de nuevos aislamientos de A. acidipropionici. Con este fin, se validó la sonda Pap446, específica de A. acidipropionici mediante ensayos de hibridación con cepas de esta especie de diferente origen, otras especies del mismo género o familia, y géneros no relacionados. Se obtuvieron ocho aislamientos con características de propionibacterias a partir de leche y heces de vacas. La sonda Pap446, hibridó únicamente con los aislamientos III y VI. La identidad de estos aislamientos fue confirmada a través de PCR con cebadores específicos para propionibacterias y para A. acidipropionici, y mediante secuenciación del ADNr 16S.
RESUMEN
Beta adrenergic receptors(beta-ARs) were detected in retina, trabecular meshwork, iris and ciliary body in eye of human, cat, rabbit, and bovine using in vitro autoradiography and majority of the beta-ARs found in eye are the beta2 subtype. Recently, the beta2-AR gene has been cloned from hamster, human, rat and monkey using molecular biological methods. Expression of beta2-AR mRNA were demonstrated in smooth muscle, kidney, ovary, brain, adipose tissue, heart, epithelial cells, thymus, lung and liver. However, studies about expression and distribution of beta2-AR mRNA in the eye have not been done yet. Author have characterized the expression of beta2-AR mRNA in rat eye using in situ hybridization with 35S-UTP riboprobe. beta2-AR mRNA was expressed in corneal epithelium and stroma, ciliary epithelium, vessels of ciliary body, choroidal vessel, and retina. In contrast it was not expressed in iris and sclera in the rat eyes. These results support the hypothesis that beta2-AR mRNA may be synthesized in the various ocular tissue and its characterized distribution suggests partially that beta2-ARs are related with aqueous production and blood supply of the eye.