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ABSTRACT Escherichia coli and Klebsiella pneumoniae are the most common pathogens causing urinary tract infections in humans and animals. Close contact between humans and companion animals can facilitate the spread of multidrug resistant pathogens between both species. The objective of the research was to characterize extended-spectrum ß-lactamases (ESBL) -producing E. coli and K. pneumoniae isolated from dogs with urinary tract infections in the metropolitan area of Valle del Aburrá (Antioquia, Colombia). Three-hundred seventy-one urine samples collected from March 2018 to March 2019 in a veterinary clinical laboratory were analyzed. E. coli and K. pneumoniae isolates were detected in chromogenic agar and identified by biochemical tests. Susceptibility testing was performed by disc diffusion and ESBL production was evaluated by the double disk test in all isolates. MIC determination of ESBL-positive isolates were performed on the automated VITEK®2 system. Multiple PCR was used for the detection of CTX-M beta-lactamases (group 1, 2, 9 and 8/25), SHV, TEM, and AmpC of plasmid origin in ESBL-positive isolates. In total 22 out 371 isolates were positive for ESBL production by double disc test, 11 E. coli (ESBL-Ec) and 11 K. pneumoniae (ESBL-Kp). The multiple PCR detected CTX-M group 1 in the 22 ESBL-positive isolates. Multi-drug resistance was observed in all ESBL-producing isolates. In conclusion, a high frequency of antibiotic multi-resistance was found in ESBL-Ec and ESBL-Kp. The main ESBL detected was CTX-M group 1, which also prevails in human isolates.
RESUMEN Escherichia coli y Klebsiella pneumoniae son los patógenos más comunes causantes de infecciones en tracto urinario en humanos y animales. El contacto estrecho con los animales de compañía puede favorecer la diseminación de patógenos multiresistentes entre ambas especies. El objetivo de la investigación fue caracterizar E. coli (Ec -BLEE) y K. pneumo-niae (Kp -BLEE) productores de betalactamasas de espectro extendido provenientes de aislados de caninos con infecciones del tracto urinario del Área Metropolitana del Valle de Aburrá. 371 muestras de orina de caninos colectadas entre marzo de 2018 y marzo 2019 en un laboratorio clínico veterinario fueron analizadas. E. coli y K. pneumoniae se detectaron en agar cromogénico y se identificaron mediante pruebas bioquímicas. La prueba de susceptibilidad se realizó por difusión en disco y la producción de BLEE se evaluó por test de doble disco en todos los aislados. La determinación de la CIM en aislados positivos a BLEE se realizó en el sistema automatizado VITEK®2. Se utilizó PCR múltiple para la detección de betalactamasas tipo CTX-M (grupo 1, 2, 9 y 8/25), SHV, TEM y AmpC de origen plasmídico en aislados positivos a BLEE. Un total de 22 de 371 aislados fueron positivos a BLEE por test de doble disco, 11 E. coli (Ec -BLEE) y 11 K. pneumoniae (Kp-BLEE). La PCR detectó CTX-M grupo 1 en los 22 aislados positivos a BLEE. Se observó multirresistencia en todos los aislamientos productores de BLEE. En conclusión, se encontró una alta frecuencia de multirresistencia en Ec-BLEE y Kp-BLEE. La principal BLEE detectada fue CTX-M grupo 1, que también predomina en aislados humanos.
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Resumen Introducción: Los procesos reflexivos e interrogativos necesarios para desarrollar preguntas de investigación cualitativas efectivas, pueden dar forma y dirección a un estudio, sin embargo; a menudo se subestiman. Las buenas preguntas de investigación no necesariamente producen una buena investigación, pero las preguntas mal concebidas o elaboradas probablemente crearán problemas que afectarán todas las etapas posteriores de un estudio. Objetivo: Este artículo aborda el desarrollo de preguntas de investigación y cómo los procesos de generación y refinación de preguntas son fundamentales para la conformación de un estudio cualitativo. Desarrollo: Generar preguntas de investigación relevantes y adecuadas requiere que el investigador preste atención no solo a las preguntas en sí mismas, sino a sus conexiones con todos los componentes del diseño; es decir, los objetivos a los que pueden responder las preguntas; las implicaciones para las preguntas relacionadas con su marco conceptual; los métodos que podría usar para responder las preguntas y las amenazas de validez que deberá abordar. Al igual que con los otros componentes del diseño, reflexionar sobre estos temas es una herramienta extremadamente útil para desarrollar sus preguntas. Conclusiones: En los estudios cualitativos, el proceso continuo de preguntas es una parte integral en la comprensión de las vidas y perspectivas de otros. Es importante enfatizar que este tipo de investigación requiere que el investigador asuma una postura epistemológica, seleccione un método y también elija ciertas técnicas que le permitan recopilar información en el trabajo de campo.
Abstract Introduction: Although frequently underestimated, reflexive and interrogative processes needed to develop the research questions in qualitative research, can give shape and direction to a study. Good research questions do not necessarily produce a good research study, but badly conceived or elaborated research questions can create problems which will further have impacts on all stages of the study. Objective: This study addresses the development of research questions and how their generation and refinement processes are fundamental in the conformation of a qualitative study. Development: Generating relevant and adequate research questions requires the researchers to give attention to the questions and their relations to all the components of the design; in other words; to the objectives, the conceptual framework, the methods used, the possible implications involved, the validity, etcetera. Reflecting on these issues is a useful tool while developing research questions. Conclusions: In qualitative studies, the ongoing process of questioning is integral to the comprehension of the lives and perspectives of others. So, it is important to emphasize that qualitative research requires the researchers to adopt an epistemological posture by selecting methods and procedures which can support their field work information gathering.
Resumo Introdução: Os processos reflexivos e interrogativos necessários para desenvolver perguntas de pesquisa qualitativas efetivas podem dar forma e direção a um estudo, porém frequentemente são subestimados. As boas perguntas de pesquisa não necessariamente produzem uma boa pesquisa, mas as perguntas mal concebidas ou elaboradas provavelmente criarão problemas que afetarão todas as etapas posteriores de um estudo. Objetivo: Este artigo aborda o desenvolvimento de perguntas de pesquisa e como os processos de produção e refinamento de preguntas são fundamentais para a conformação de um estudo qualitativo. Desenvolvimento: Gerar perguntas de pesquisa relevantes e adequadas requer que o pesquisador preste atenção não só às preguntas em si mesmas, senão a suas conexões com todos os componentes do desenho: quer dizer, os objetivos aos que podem responder as preguntas; as implicações para as preguntas relacionadas com seu marco conceptual; os métodos que poderia usar para responder as perguntas e as ameaças de validade que deverá abordar. Ao igual que com os outros componentes do desenho, refletir sobre estes temas é uma ferramenta extremadamente útil para desenvolver suas perguntas. Conclusões: Nos estudos qualitativos, o processo contínuo de perguntas é uma parte integral na compreensão das vidas e perspectivas de outros. É importante enfatizar que este tipo de pesquisa, requer que o pesquisador assuma uma postura epistemológica, selecione um método e também eleja certas técnicas que lhe permitam recopilar informação no trabalho de campo.
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Cell wall pectins are some of the most complex biopolymers known, and yet their functions remain largely mysterious. The aim of this paper was to deepen the study of the spatial pattern of pectin distribution in the aperture of Oenothera hookeri.velans ster/+ster fertile pollen. We used "in situ" immunocytochemical techniques at electron microscopy, involving monoclonal antibodies JIM5 and JIM7 directed against pectin epitopes in fertile pollen grains of Oenothera hookeri.velans ster/+ster. The same region was also analyzed by classical cytochemistry for polysaccharide detection. Immunogold labelling at the JIM7 epitope showed only in mature pollen labelling mainly located at the intine endo-aperture region. Cytoplasmic structures near the plasma membrane of the vegetative cell showed no labelling gold grains. In the same pollen stge the labelling at the JIM5 epitope was mostly confined to a layer located in the limit between the endexine and the ektexine at the level of the border of the oncus. Some tubuli at the base of the ektexine showed also an accumulation of gold particles. No JIM5 label was demonstrated in the aperture chamber and either in any cytoplasmic structure of the pollen grains. The immunocytochemical technique, when compared with the traditional methods for non-cellulose polysaccharide cytochemistry is fare more sensitive and allows the univocal determination of temporal and spatial location of pectins recognized by the JIM7 and JIM5 MAbs.