In the present study, a nested-PCR system, targeting the TbD1 region, involving the performance of conventional PCR followed by real-time PCR, was developed to detect Mycobacterium bovis in bovine/bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. In terms of analytical sensitivity, the DNA of M. bovis AN5 was detected up to 1.56 ng with conventional PCR, 97.6 pg with real-time PCR, and 1.53 pg with nested-PCR in the reaction mixture. The nested-PCR exhibited 100% analytical specificity for M. bovis when tested with the DNA of reference strains of environmental mycobacteria and closely-related Actinomycetales. A clinical sensitivity value of 76.0% was detected with tissue samples from animals that exhibited positive results in the comparative intradermal tuberculin test (CITT), as well as from those with lesions compatible with tuberculosis (LCT) that rendered positive cultures. A clinical specificity value of 100% was detected with tissue samples from animals with CITT- results, with no visible lesions (NVL) and negative cultures. No significant differences were found between the nested-PCR and culture in terms of detecting CITT+ animals with LCT or with NVL. No significant differences were recorded in the detection of CITT- animals with NVL. However, nested-PCR detected a significantly higher number of positive animals than the culture in the group of animals exhibiting LCT with no previous records of CITT. The use of the nested-PCR assay to detect M. bovis in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.
OBJETIVO: Avaliar o perfil epidemiológico de pacientes com traumatismo raquimedular atendidos em hospital terciário. MÉTODOS: Estudo descritivo, transversal, prospectivo, com 321 pacientes vítimas de traumatismo raquimedular, realizado de janeiro de 2008 a junho de 2012. Foram estudadas as variáveis: sexo; idade; estado civil; profissão; escolaridade; religião; procedência; etiologia, morfologia e região da lesão; condição neurológica pela escala da ASIA e lesões associadas. RESULTADOS: Amostra constituída por 72% pacientes do sexo masculino e 28% do feminino, prevalência da faixa etária de 21 a 30 anos. Os estados civis mais frequentes foram união estável (46,8%) e solteiros (41,7%). O nível de escolaridade foi ensino fundamental incompleto (57%) e completo (17,8%). As causas mais frequentes foram acidentes automobilísticos (38,9%) e queda (27,4%). A lesão mais presente foi fratura explosão (23,7%), as regiões mais afetadas foram cervical subaxial (41,7%) e transição toracolombar (30,5%). A lesão associada mais frequente foi traumatismo cranioencefálico (TCE) (28,2%). O estado neurológico mais observado na internação/alta foi ASIA-E. Ocorreram 25 óbitos (7,8%), sendo que 76% com lesão na região cervical foram estratificados com ASIA-A, e 68% tiveram complicações respiratórias. CONCLUSÃO: O trauma raquimedular acometeu mais adultos jovens do sexo masculino com união estável e baixo nível de escolaridade. A causa mais frequente foi acidente automobilístico, o tipo de lesão foi fratura explosão e a região cervical a mais acometida. A condição neurológica mais presente foi ASIA-E, o TCE foi a lesão associada mais frequente e a maior gravidade pela classificação da ASIA nos casos de envolvimento cervical aumentou o risco de complicações respiratórias e morbidade e mortalidade.
Vaccinia virus (VACV) has had an important role for humanity because of its use during the smallpox eradication campaign. VACV is the etiologic agent of the bovine vaccinia (BV), an emerging zoonosis that has been associated with economic, social, veterinary and public health problems, mainly in Brazil and India. Despite the current and historical VACV importance, there is little information about its circulation, prevalence, origins and maintenance in the environment, natural reservoirs and diversity. Brazilian VACV (VACV-BR) are grouped into at least two groups based on genetic and biological diversity: group 1 (G1) and group 2 (G2). In this study, we went to the field and investigated VACV clonal diversity directly from exanthemous lesions, during BV outbreaks. Our results demonstrate that the G1 VACV-BR were more frequently isolated. Furthermore, we were able to co-detect the two variants (G1 and G2) in the same sample. Molecular and biological analysis corroborated previous reports and confirmed the co-circulation of two VACV-BR lineages. The detected G2 clones presented exclusive genetic and biological markers, distinct to reference isolates, including VACV-Western Reserve. Two clones presented a mosaic profile, with both G1 and G2 features based on the molecular analysis of A56R, A26L and C23L genes. Indeed, some SNPs and INDELs in A56R nucleotide sequences were observed among clones of the same virus population, maybe as a result of an increased mutation rate in a mixed population. These results provide information about the diversity profile in VACV populations, highlighting its importance to VACV evolution and maintenance in the environment.
Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.
MRI was significantly superior to CT in the diagnosis of bone swelling, PLC injury, disk herniation, spinal canal compression, spinal cord contusion and swelling present in SCT. MRI detected a larger number of lesions than CT and is highly useful for the diagnosis of soft tissue and intrathecal injuries.
Orf virus is the etiological agent of contagious ecthyma, a severe exanthematic disease that affects small ruminants. Orf virus is zoonosis that is associated with occupational contact with infected animals in human disease. Clinically, contagious ecthyma is characterized by the appearance of vesicles, pustules, ulcers, and papillomatous proliferative lesions on the skin of the lips and nostrils. Here we describe a case of lethal cutaneous multifocal Orf virus infection in goats in the Amazon region of Brazil. Exanthematic lesions were collected and epidemiological and clinical data were obtained. Orf virus was detected using PCR amplification of the whole B2L, VIR, and VEGF open reading frame. Phylogenetic analysis revealed that this virus clustered together with the Orf virus samples isolated during classical contagious ecthyma. The present work is the first to report a severe proliferative Orf virus case in South America.
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