RESUMEN
Forty-two recent (1997-1999) Spanish isolates of Francisella tularensis subsp.holarctica were tested in a broth microdilution method for their susceptibilities to 29 antimicrobial agents, including penicillins, cephalosporins, cephamicins, monobactams, penems, aminoglycosides, tetracyclines, macrolides, quinolones, chloramphenicol and fosfomycin. The isolates were resistant to beta-lactam antibiotics and susceptible to chloramphenicol, ciprofloxacin, levofloxacin and norfloxacin.
Asunto(s)
Antibacterianos/farmacología , Francisella tularensis/efectos de los fármacos , Animales , Arvicolinae/microbiología , Francisella tularensis/aislamiento & purificación , Liebres/microbiología , Humanos , Pruebas de Sensibilidad Microbiana/veterinaria , Garrapatas/microbiologíaAsunto(s)
Brotes de Enfermedades/veterinaria , Francisella tularensis/clasificación , Tularemia/veterinaria , Animales , Arvicolinae , Francisella tularensis/aislamiento & purificación , Francisella tularensis/patogenicidad , Humanos , Conejos , España/epidemiología , Garrapatas , Tularemia/epidemiología , Tularemia/etiología , VirulenciaRESUMEN
On the basis of a species-specific PCR assay, a RFLP analysis for typing of Haemophilus parasuis strains was developed and evaluated. Amplification was based on the gene tbpA, encoding a transferrin-binding protein. RFLP analysis of the 1.9-kb tbpA-amplicon using TaqI, AvaI and RsaI endonucleases produced 12 different patterns for the reference strains of the 15 known H. parasuis serovars, and showed a high heterogeneity (33 RFLP groups) for 101 H. parasuis clinical isolates tested. The sensitivity, typeability (100% versus 65% for immunodiffusion), high degree of discrimination (0.93 versus 0.84 for immunodiffusion), simplicity and low cost per test make this PCR-RFLP assay a useful method for typing H. parasuis and, therefore, for studying the epidemiology of outbreaks of Glässer's disease.
Asunto(s)
Haemophilus/genética , Reacción en Cadena de la Polimerasa/veterinaria , Proteína A de Unión a Transferrina/genética , Animales , ADN Bacteriano/química , ADN Bacteriano/genética , Haemophilus/química , Haemophilus/clasificación , Infecciones por Haemophilus/microbiología , Infecciones por Haemophilus/veterinaria , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Sensibilidad y Especificidad , Porcinos , Enfermedades de los Porcinos/microbiología , Proteína A de Unión a Transferrina/químicaRESUMEN
We evaluated three molecular methods for identification of Francisella strains: pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphism (AFLP) analysis, and 16S rRNA gene sequencing. The analysis was performed with 54 Francisella tularensis subsp. holarctica, 5 F. tularensis subsp. tularensis, 2 F. tularensis subsp. novicida, and 1 F. philomiragia strains. On the basis of the combination of results obtained by PFGE with the restriction enzymes XhoI and BamHI, PFGE revealed seven pulsotypes, which allowed us to discriminate the strains to the subspecies level and which even allowed us to discriminate among some isolates of F. tularensis subsp. holarctica. The AFLP analysis technique produced some degree of discrimination among F. tularensis subsp. holarctica strains (one primary cluster with three major subclusters and minor variations within subclusters) when EcoRI-C and MseI-A, EcoRI-T and MseI-T, EcoRI-A and MseI-C, and EcoRI-0 and MseI-CA were used as primers. The degree of similarity among the strains was about 94%. The percent similarities of the AFLP profiles of this subspecies compared to those of F. tularensis subsp. tularensis, F. tularensis subsp. novicida, and F. philomiragia were less than 90%, about 72%, and less than 24%, respectively, thus permitting easy differentiation of this subspecies. 16S rRNA gene sequencing revealed 100% similarity for all F. tularensis subsp. holarctica isolates compared in this study. These results suggest that although limited genetic heterogeneity among F. tularensis subsp. holarctica isolates was observed, PFGE and AFLP analysis appear to be promising tools for the diagnosis of infections caused by different subspecies of F. tularensis and suitable techniques for the differentiation of individual strains.
Asunto(s)
Técnicas de Tipificación Bacteriana , Francisella tularensis/clasificación , Polimorfismo de Longitud del Fragmento de Restricción , Tularemia/diagnóstico , Animales , ADN Ribosómico/análisis , Electroforesis en Gel de Campo Pulsado , Francisella tularensis/genética , Genes de ARNr , Genotipo , Humanos , Datos de Secuencia Molecular , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , España/epidemiología , Tularemia/microbiologíaRESUMEN
Pasteurella canis biotype 1, usually associated with the oral cavity of dogs and cats, or with human wound infections following dog bites, was isolated from newborn puppies with a fatal systemic infection. The identity of P. canis was confirmed by arbitrarily primed polymerase chain reaction and the organism was susceptible to all the penicillins, cephalosporins, tetracyclines and fluoroquinolones tested and to most of the aminoglycosides tested. This represents the first report of systemic pasteurellosis caused by P. canis in dogs.
Asunto(s)
Enfermedades de los Perros/microbiología , Infecciones por Pasteurella/microbiología , Pasteurella/aislamiento & purificación , Animales , Animales Recién Nacidos , Antibacterianos/farmacología , Sangre/microbiología , ADN Bacteriano/genética , Enfermedades de los Perros/patología , Perros , Femenino , Hígado/microbiología , Pulmón/microbiología , Masculino , Pruebas de Sensibilidad Microbiana , Pasteurella/efectos de los fármacos , Pasteurella/genética , Infecciones por Pasteurella/patología , Especificidad de la Especie , Bazo/microbiología , Tráquea/microbiologíaRESUMEN
Actinobacillus seminis isolates were cultured from the semen (two isolates) and the left testis (one isolate) of two one-year-old rams in León, Spain. One animal showed lesions of chronic unilateral orchitis and epididymitis while the other appeared to suffer a subclinical infection and only a moderate number of pleomorphic rods and inflammatory cells were present in its semen. The isolates were biochemically similar to the A. seminis type strain NCTC 10851 and two other European A. seminis isolates, except that they produced acid from sorbitol; their identity was confirmed by arbitrarily primed polymerase chain reaction. The isolates were also tested against 30 antimicrobial agents, and only marbofloxacin was found active against all of them. As far as is known, this is the first report of A. seminis isolation from rams in southern Europe.