Comparación entre hibridización in situ e inmunoperoxidasa indirecta para la determinación de la ubicación de pasteurella multocida en conejos afectados con enfermedad respiratoria
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Franco Mesa, Martha Lilia
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Trabajo de grado - Maestría
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EspañolFecha de publicación
2012
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La Pasteurella multocida es considerada un habitante normal del tracto respiratorio superior de los conejos y es el patógeno más frecuentemente aislado en la enfermedad respiratoria de esta especie, sin embargo poco se conoce acerca de su ubicación, distribución y densidad. En el presente estudio se estandarizó la técnica de hibridización in situ (HIS) para P. multocida, y se comparó esta marcación con la obtenida con técnicas de rutina como Inmunoperoxidasa Indirecta (IPI), confirmando así la presencia de la bacteria en fosa nasal de conejos afectados con enfermedad respiratoria en su forma natural, tanto en conejos asintomáticos de 1-21 días de edad como en conejos enfermos con síntomas de rinitis y septicemia, mediante el uso de una sonda de oligonucleótidos marcada con digoxigenina y dirigida al 16S rRNA. HIS permitió demostrar la ubicación preferencial de esta bacteria en células ciliadas, espacios interepiteliales y células caliciformes.
Abstract. Pasteurella multocida is considered to be a normal inhabitant of the upper respiratory tract of rabbits and the pathogen most commonly isolated in respiratory disease of this species, however little is known about its location, distribution, and density. In this study an in situ hybridization (ISH) technique for P. multocida was standardized, and its results were compared with routine techniques such as indirect Immunoperoxidase (IPI), thus confirming the presence of the bacteria in the nasal cavity of rabbits affected by respiratory disease in its natural form, both in asymptomatic 1-21 day old rabbits and in sick animals with symptoms of rhinitis and septicemia, an oligonucleotide probe directed towards the 16S gen of the rRNA labeled with digoxigenin was used. ISH demonstrated the preferential location of this bacterium on ciliated cells, interepithelial spaces and goblet cells.
Abstract. Pasteurella multocida is considered to be a normal inhabitant of the upper respiratory tract of rabbits and the pathogen most commonly isolated in respiratory disease of this species, however little is known about its location, distribution, and density. In this study an in situ hybridization (ISH) technique for P. multocida was standardized, and its results were compared with routine techniques such as indirect Immunoperoxidase (IPI), thus confirming the presence of the bacteria in the nasal cavity of rabbits affected by respiratory disease in its natural form, both in asymptomatic 1-21 day old rabbits and in sick animals with symptoms of rhinitis and septicemia, an oligonucleotide probe directed towards the 16S gen of the rRNA labeled with digoxigenin was used. ISH demonstrated the preferential location of this bacterium on ciliated cells, interepithelial spaces and goblet cells.