Direct Detection of Mycobacterium bovis in Bovine Lymph Nodes by PCR

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Occurrence of mycobacteria in bovine milk samples from both individual and collective bulk tanks at farms and informal markets in the southeast region of Sao Paulo, Brazil

Background: Mycobacterium spp. is one of the most important species of zoonotic pathogens that can be transmitted from cattle to humans. The presence of these opportunistic, pathogenic bacteria in bovine milk has emerged as a public-health concern, especially among individuals who consume raw milk and related dairy products. To address this concern, the Brazilian control and eradication program focusing on bovine tuberculosis, was established in 2001. However, bovine tuberculosis continues to afflict approximately 1,3 percent of the cattle in Brazil. In the present study, 300 samples of milk from bovine herds, obtained from both individual and collective bulk tanks and informal points of sale, were cultured on Löwenstein-Jensen and Stonebrink media. Polymerase chain reaction (PCR)-based tests and restriction-enzyme pattern analysis were then performed on the colonies exhibiting phenotypes suggestive of Mycobacterium spp., which were characterized as acid-fast bacilli.Results: Of the 300 bovine milk samples that were processed, 24 were positively identified as Mycobacterium spp.Molecular identification detected 15 unique mycobacterial species: Mycobacterium bovis, M. gordonae, M. fortuitum, M. intracellulare, M. flavescens, M. duvalii, M. haemophilum, M. immunogenum, M. lentiflavum, M. mucogenicum, M. novocastrense, M. parafortuitum, M. smegmatis, M. terrae and M. vaccae. The isolation of bacteria from the various locations occurred in the following proportions: 9 percent of the individual bulk-tank samples, 7 percent of the collective bulk-tank samples and 8 percent of the informal-trade samples. No statistically significant difference was observed between the presence of Mycobacterium spp. in the three types of samples collected, the milk production profiles, the presence of veterinary assistance and the reported concerns about bovine tuberculosis prevention in the herds.Conclusion: The microbiological cultures associated with PCR-based identification tests are possible tools for the investigation of the presence of Mycobacterium spp. in milk samples. Using these methods, we found that the Brazilian population may be regularly exposed to mycobacteria by consuming raw bovine milk and related dairy products. These evidences reinforces the need to optimize quality programs of dairy products, to intensify the sanitary inspection of these products and the necessity of further studies on the presence of Mycobacterium spp. in milk and milk-based products. © 2013 Franco et al.; licensee BioMed Central Ltd.

Multiplex-PCR for differentiation of Mycobacterium bovis from Mycobacterium tuberculosis complex

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Tuberculosis: challenges to improve the treatment

Tuberculosis is one of the leading causes of morbidity and mortality worldwide. Current treatment has several challenges including multi-drug resistance, extensively drug-resistance and HIV co-infection. Problems related to patients, treatment and health care system also contributes negatively to this panel. This review summarizes the main obstacles to treat tuberculosis and discuss several strategies to improve the treatment.

Prevalência de reagentes ao teste de tuberculinização no rebanho bovino leiteiro da região de São Carlos - SP

Tuberculosis (TB), an infectious disease readily transmitted between animals and humans, is an great threat to public and animal health a like and has been declared an international emergency. According to the WHO, animal TB is a source of much concern, especially in the poorer countries where there is little awareness of the problem. Thus, improvements in veterinary health are essential, with regard to infection by Mycobacterium bovis, particularly in areas where the public is at risk. The prevalence of infected animals in Brazil is about 1%, whereas, in the dairy cattle region of São Carlos, the average proportion of positive tests was found to be 3. 1% from 1987 to 1996. The current research was carried out to investigate the prevalence, in the region, of dairy cattle reacting positively to the tuberculin test, from 1997 to the first half of 2001. The results were obtained by searching through the data collected by the Federal Inspection Authority, and they indicated an average prevalence of 1.3% over the period, similar to the official national average. It was concluded that bovine TB continues to be present in the region, posing a potential threat of infection to the human population.

Ocorrência da brucelose e tuberculose bovina e percepção de riscos de produtores de leite do município de Paranaíba, Mato Grosso do Sul: Gilson Luiz Piva Filho. -

Pós-graduação em Ciência Animal - FMVA

Incidência de linfoadenopatia compatível com tuberculose em bovinos abatidos em frigorífico de exportação no período de 2007 a 2011

Pós-graduação em Medicina Veterinária - FMVZ

Ocorrência de micobactérias em amostras de leite bovino provenientes de tanques de expansão individuais e coletivos de propriedades rurais e do comércio informal na região sudeste do estado de São Paulo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Resposta imune à infecção por Mycobacterium bovis em linhagens de camundongos geneticamente selecionados (Seleção IV-A)

Pós-graduação em Medicina Veterinária - FMVZ

Análise histomorfológica e molecular de lesões granulomatosas sugestivas de tuberculose bovina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparação entre métodos diagnósticos da tuberculose em bovinos abatidos em matadouros-frigoríficos do Estado de São Paulo

Pós-graduação em Medicina Veterinária - FCAV

Caracterização histomorfológica de lesões granulomatosas sugestivas de tuberculose ganglionar e pulmonar em bovinos abatidos na região noroeste do estado de SãoPaulo

Tuberculosis infection among humans transmitted by products of animal origin can be caused by Mycobacterium bovis, a concern in developing countries, because the number of human cases is relatively high compared with countries that have implemented programs to eradicate bovine tuberculosis for many years. Because it is a chronic disease with subclinical evolution, it is transmitted to other animals in the herd livestock resulting in high loss losses resulting high (10 to 25%), as well as the zoonotic nature of the disease on public health. This work aimed to study the histomorphology of granulomas in samples of lymph nodes and lungs of animals with suggestive lesions of tuberculosis, as well as those similar to the sanitary inspection. The animals clinically healthy ante-mortem, were slaughtered in a slaughterhouse in the northwestern state of Sao Paulo between April 2008 and April 2009, considering that carcasses had lymph nodes with lesions typical of tuberculosis, exhibiting nodular and hemorrhagic appearance. The caseous or calcified lesions, purulent or not, of various sizes and shapes, were also evaluated. Of the 307,661 animals slaughtered in the mentioned period, 494 gross suggestive lesions of tuberculosis and from 29 cities in the state of Sao Paulo, 16 from Mato Grosso do Sul, Goias 15, one from Mato Grosso and Paraná. Among these, 200 samples (40,5%) were processed for histopathological evaluation by hematoxylin-eosin (HE) and 90 (45%) were subjected to Ziehl-Neelsen (ZN) and examined by light microscopy. In HE staining, histopathological changes of tuberculosis were found in 197 (98,5%), classified according to the stage of development adopted by Wango et al.(31), where 29 samples were included in stage I (14,5%), 53 in stage II (26,5%), 87 in stage III (43,5%), 28 in stage IV (14%). In the ZN staining Mycobacterium sp was found in 89 (99%) of the specimens. The rate of tuberculosis in the herd studied was 0,16%. We conclude that the identification of the disease in slaughterhouses and meat companies from significant gross lesions, enables the implementation of effective measures, specifically the origin of the disease in cattle herds, providing control measures to prevent its spread.

On the interaction of bovine serum albumin with ionic surfactants: Temperature induced EPR changes of a maleimide nitroxide reflect local protein dynamics and probe solvent accessibility

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Functional characterization by genetic complementation of aroB-Encoded dehydroquinate synthase from Mycobactetium tuberculosis H37Rv and its heterologous expression and purification

The recent recrudescence of Mycobacterium tuberculosis infection and the emergence of multidrug-resistant strains have created an urgent need for new therapeutics against tuberculosis. The enzymes of the shikimate pathway are attractive drug targets because this route is absent in mammals and, in M. tuberculosis, it is essential for pathogen viability. This pathway leads to the biosynthesis of aromatic compounds, including aromatic amino acids, and it is found in plants, fungi, bacteria, and apicomplexan parasites. The aroB-encoded enzyme dehydroquinate synthase is the second enzyme of this pathway, and it catalyzes the cyclization of 3-deoxy-D-arabino-heptulosonate-7-phosphate in 3-dehydroquinate. Here we describe the PCR amplification and cloning of the aroB gene and the overexpression and purification of its product, dehydroquinate synthase, to homogeneity. In order to probe where the recombinant dehydroquinate synthase was active, genetic complementation studies were performed. The Escherichia coli AB2847 mutant was used to demonstrate that the plasmid construction was able to repair the mutants, allowing them to grow in minimal medium devoid of aromatic compound supplementation. In addition, homogeneous recombinant M. tuberculosis dehydroquinate synthase was active in the absence of other enzymes, showing that it is homomeric. These results will support the structural studies with M. tuberculosis dehydroquinate synthase that are essential for the rational design of antimycobacterial agents.

Influence of the season of year, stage of lactation and milking time on the chloride content in bovine milk

From January to October 1995, chloride content determinations were accomplished in 2,218 samples of milk collected from 67 quarters of 17 lactating cows, in initial, middle and final stages of lactation, in the morning and afternoon milking. The highest means of chloride contents were observed among the milk samples collected in the final stage of lactation (0,203 gC1/100ml), in the summer (0,212 gC1/100ml) and in the morning milking (0,189 g C1/100ml). The differences observed between the chloride contents in the samples obtained in the different stages of lactation and the seasons of the year were statistically significant (P<0.0001). The gratest occornce of chloride at rates superior to 0.22 gC1100ml was observed in the final stge of lactation (31.4%), summer (44.2%) and after the morning milking (27.8%). These results evidentiate the influence of physiological and external factors (stage of lactation and season of the year) on the concentration of chloride of milk.