RESUMEN
Two primer set/probe combinations targeting variable regions on the 23S rRNA gene were designed to detect and quantify chlamydiae in DNA extracted from brain swabs of the endangered Houston toad (Anaxyrus houstonensis) using SYBRGreen- and Taqman-based quantitative polymerase chain reaction (qPCR). Prevalence and abundance values for samples were generally different between SYBRGreen- and Taqman-based detection methods, with higher specificity observed for Taqman-based detection. Of the 314 samples analyzed, initial screening with SYBRGreen-based qPCR retrieved 138 positive samples, of which 52 were confirmed by Taqman-based analyses as chlamydiae. All of these samples were subsequently identified as Chlamydia pneumoniae by specific qPCR and confirmed by comparative sequence analyses of 23S rRNA gene amplicons. These results demonstrate the usefulness of our developed qPCR methods to screen for and verify prevalence of chlamydiae in DNA of brain swabs, and ultimately specifically identify and quantify chlamydiae, specifically C. pneumoniae in these samples.
Asunto(s)
Encéfalo , Bufonidae , Chlamydiaceae , Bufonidae/microbiología , Animales , Especies en Peligro de Extinción , Encéfalo/microbiología , Chlamydiaceae/aislamiento & purificaciónRESUMEN
Illumina-based 16S rRNA V3 amplicon sequencing of total DNA obtained from soft tissue lesions (joint granulomas) of the endangered Houston toad (Anaxyrus houstonensis) demonstrated that many reads represented members of the actinobacterial Mycobacterium chelonae-abscessus complex. In order to quantify members of this complex in those lesions, we designed three complex-specific primer set/probe combinations (sets I, II and III) targeting variable regions on the 23S rRNA gene for SybrGreen- and Taqman-based quantitative polymerase chain reaction (qPCR). Both SybrGreen- and Taqman-based analyses specifically detected members of the M. chelonae-abscessus complex in lesion samples, with numbers between 104 and 107 cells per 100-mg sample. Values within individual samples were generally comparable between SybrGreen- and Taqman-based detection methods and between all primer set/probe combinations, except for SybrGreen-based analyses of a few samples analyzed with primer set I that used a less specific forward primer. The development of highly specific detection and quantification methods for members of the M. chelonae-abscessus complex in lesion samples can enable group specific tracking of these organisms, particularly in captive or stewardship settings where source and transmission monitoring are valuable tools to husbandry and species conservation.
Asunto(s)
Mycobacterium abscessus , Mycobacterium chelonae , Mycobacterium abscessus/genética , Mycobacterium chelonae/genética , Filogenia , ARN Ribosómico 16S/genética , ARN Ribosómico 23S/genéticaRESUMEN
Head-starting of the federally endangered Houston toad (Anaxyrus houstonensis), that is, the release of egg strands, tadpoles, and metamorphic juveniles produced in captivity into the original breeding ponds, requires assessment of potential threats for the transmission of pathogens from captive to free-ranging toads. We used Illumina-based 16S rRNA V3 amplicon sequencing to investigate the community structure of bacteria from skin lesions of captive Houston toad and habitat (pond) samples. Proteobacteria, alone or together with Actinobacteria and, in some samples, Cyanobacteria represented virtually all reads in tissue lesion samples, whereas pond samples were much more diverse, with Acidobacteria, Actinobacteria, Bacteriodetes, Chloroflexi, Cyanobacteria, Firmicutes, Planctomycetes, Proteobacteria, and Verrucomicrobia present with little variation between samples. If present in lesions, Actinobacteria were largely represented by Mycobacteriaceae, and here mainly by one sequence identical to sequences of members of the Mycobacterium chelonae-abscessus complex. In pond samples, mycobacteria represented only a small portion of the actinobacteria, although at higher diversity with six distinct reads. Sequences for reads obtained from pond samples were identical to those representing the M. chelonae-abscessus complex, a group with Mycobacterium marinum, Mycobacterium kansasii, Mycobacterium avium, a group with Mycobacterium vaccae, Mycobacterium fortuitum, Mycobacterium poriferae, and a group with Mycobacterium elephantis and Mycobacterium celeriflavum, whereas sequences of high similarity were detected for reads related to those of Mycobacterium holsaticum, Mycobacterium pallens, and Mycobacterium obuense, and Mycobacterium goodii. Our results indicated that lesions observed on the Houston toad in captivity are not the result of mycobacteria in every case, and that the presence of mycobacteria in the captive colony does not represent a novel pathogen threat to the wild populations because such bacteria are also seen in the natural pond habitats for the Houston toad.
Asunto(s)
Mycobacterium , Animales , Ecosistema , Mycobacteriaceae , Mycobacterium/genética , ARN Ribosómico 16S/genéticaRESUMEN
Objetivo: gestionar la creación y puesta en marcha de una red de apoyo intersectorial, cuyo fin es brindar soporte y contribuir a la solución de la problemática de las familias. Materiales y Métodos: estudio descriptivo, llevado a cabo en 42 familias, 15 líderes comunitarios y representantes de 8 instituciones y 11 trabajadores del centro de salud de un barrio de estratos 1 y 2 de la ciudad de Bucaramanga. Se aplicó un formato de valoración por dominios de la Asociación Norteamericana de Diagnósticos de Enfermería (NANDA) para identificar la problemática de las familias. El procesamiento de los datos, se realizó en Excel mediante la obtención de frecuencias y promedios para su posterior análisis. Resultados: se logró la creación y puesta en marcha de la red de apoyo, conformada por ocho instituciones del sector,con la cual se abordaron problemáticas como: inasistencia a programas de promoción y prevención, drogadicción, violencia intrafamiliar, embarazos en adolescentes. Conclusiones: la articulación interinstitucional y la conformación de redes de apoyo logran mejores efectos a nivel económico, social, espiritual, físico y psicológico, contribuyendo a la salud familiar. Asimismo, contribuye al desarrollo personal, profesional, liderazgo y construcción del conocimiento, propios de enfermería familiar como eje de abordaje en la salud pública.
Goal: To manage the creation and implementation of a network of cross sector support, which aims to provide support and contribute to the solution of the problems of families. Materials and Methods: A descriptive study was conducted in 42 families, 15 community leaders and representatives of eight institutions and 11 health center workers in a district of 1 and 2 statuses in the city of Bucaramanga. It was used the Format for valuation domains of the American Association of Nursing Diagnoses (NANDA) to identify the problems of families. The data processing was performed in Excel by obtaining frequencies and averages for further analysis. Results: Absence from promotion and prevention programs, drug addiction, domestic violence, teen pregnancy: the creation and implementation of the support network, consisting of eight institutions in the sector, which addressed issues as was achieved. Conclusions: interagency coordination and the creation of support networks achieve better effects in economic, social, spiritual, physical and psychological level, contributing to family health. It also contributes to personal, professional, leadership and knowledge. building, typical of family nursing as axis approach to public health.
Objetivo: Gerenciar a criação e implementação de uma rede de apoio intersetorial, que visa dar apoio e contribuir para a solução dos problemas das famílias. Materiais e Métodos: Estudo descritivo, realizado em 42 famílias, 15 líderes comunitários e representantes de oito instituições e 11 trabalhadores de centros de saúde no distrito de níveis 1 e 2, na cidade de Bucaramanga. Formato foi utilizada para domínios de avaliação da Associação Americana de Diagnósticos de Enfermagem (NANDA) para identificar os problemas das famílias. O processamento dos dados foi realizada no Excel através da obtenção de freqüências e médias para análise posterior. Resultados: Ausência de programas de promoção e prevenção, toxicodependência, violência doméstica, gravidez na adolescência: a criação e implementação da rede de apoio, constituída por oito instituições do setor, que abordou temas como foi conseguido. Conclusões: a coordenação interinstitucional ea criação de redes de apoio alcançar melhores efeitos em nível econômico, social, espiritual, física e psicológica, contribuindo para a saúde da família. Também contribui para a pessoal, profissional, liderança e construção do conhecimento, típico de enfermagem de família como eixo abordagem à saúde pública.