11 research outputs found
ANATOMOPATOLOGIA DA PARATUBERCULOSE EM BOVINOS LEITEIRO DE RESENDE- R.J
A Paratuberculose, uma doença causada pelo Mycobacterium avium subsp. paratuberculosis (Map), é caracterizada por uma enterite crônica granulomatosa. Visamos registrar a anatomopatologia de três casos de paratuberculose em bovinos leiteiros autóctones oriundos de Resende-RJ. Esses animais apresentavam sinais clÃnicos de paratuberculose e, após teste de ELISA positivo, foram eutanasiados e necropsiados. Amostras de intestino, linfonodos mesentéricos e válvula ileocecal foram colhidas, fixadas em formol neutro tamponado a 10%, processadas e coradas pelo HE (hematoxilina-eosina) e ZN (Ziehl-Neelsen). Alterações macroscópicas, como espessamento da parede do intestino delgado, mucosas congestas e corrugadas, foram observadas. A linfadenomegalia e linfangectasia também foram evidentes, assim como a congestão e edemaciação das válvulas ileocecais. Os principais achados histopatológicos foram enterite, linfangite e linfadenite granulomatosas. As lesões intestinais se resumiam à mucosa e submucosa, principalmente do jejuno e do Ãleo e caracterizavam-se por infiltrado inflamatório de linfócitos, eosinófilos e macrófagos epitelióides. A coloração de ZN demonstrou bacilos álcool ácido resistentes (BAAR) nos macrófagos, no ápice das vilosidades e lâmina própria e, ainda, no parênquima dos linfonodos mesentéricos. A anatomopatologia foi caracterÃstica e considerada uma valiosa ferramenta no diagnóstico da paratuberculose.
PALAVRAS-CHAVE: anatomopatológico; bovino; Map; paratuberculos
Parasitic Ulcerous Caseous Gastroesophagitis Associated with Rameshwarotrema uterocrescens
Apa antigen of Mycobacterium avium subsp. paratuberculosis as a target for species-specific immunodetection of the bacteria in infected tissues of cattle with paratuberculosis
Comparative genomics of Mycobacterium spp. have revealed conservative genes and respective proteins differently expressed in mycobacteria that could be used as targets for the species-specific immunodiagnostics. The alanine and proline-rich antigen Apa is a mycobacterial protein that present significant variability in primary sequence length and composition between members of M. avium and M. tuberculosis complexes. In this study, the recombinant Apa protein encoded by the MAP1569/ModD gene of M. avium subsp. paratuberculosis (Map) was used to generate a panel of monoclonal antibodies which were shown to recognize the most important veterinary pathogens of the M. avium complex, specifically Map and M. avium subsp. hominissuis, and which did not cross-react with M. bovis or M. tuberculosis. The produced antibodies were demonstrated to be a useful tool for the species-specific immunofluorescence or immunohistochemical detection of Map in experimentally infected cell cultures or intestinal tissues from cattle with bovine paratuberculosis and, additionally, they may be employed for the discrimination of pathogenic M. avium subspecies via Western blotting.Fil: Souza, Giliane S.. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; BrasilFil: Rodrigues, Ana Bárbara F.. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; BrasilFil: Gioffré, Andrea Karina. Instituto Nacional de TecnologÃa Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de BiotecnologÃa; Argentina. Consejo Nacional de Investigaciones CientÃficas y Técnicas; ArgentinaFil: Romano, Maria Isabel. Instituto Nacional de TecnologÃa Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de BiotecnologÃa; Argentina. Consejo Nacional de Investigaciones CientÃficas y Técnicas; ArgentinaFil: Carvalho, Eulógio C. Q.. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; BrasilFil: Ventura, Thatiana L. B.. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; BrasilFil: Lasunskaia, Elena B.. Universidade Estadual Do Norte Fluminense Darcy Ribeiro; Brasi
Mycobacterium tuberculosis Strains of the Modern Sublineage of the Beijing Family Are More Likely To Display Increased Virulence than Strains of the Ancient Sublineage
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Previous issue date: 2014Universidade Estadual do Norte Fluminense.Laboratório de Morfologia Animal e Patologia. Laboratório de Biologia do Reconhecimento. Campos, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular Aplicada a Micobactérias. Rio de Janeiro, RJ, Brasil.Universidade de São Paulo. Instituto de Ciências Biomédicas. Departamento de Imunologia. São Paulo, SP, BrasilUniversidade Estadual do Norte Fluminense.Laboratório de Morfologia Animal e Patologia. Laboratório de Biologia do Reconhecimento. Campos, RJ, Brasil.Universidade Estadual do Norte Fluminense.Laboratório de Morfologia Animal e Patologia. Laboratório de Biologia do Reconhecimento. Campos, RJ, Brasil.Universidade Estadual do Norte Fluminense.Laboratório de Morfologia Animal e Patologia. Laboratório de Biologia do Reconhecimento. Campos, RJ, Brasil.Universidade Estadual do Norte Fluminense.Laboratório de Morfologia Animal e Patologia. Laboratório de Biologia do Reconhecimento. Campos, RJ, Brasil.Universidade Estadual do Norte Fluminense.Laboratório de Morfologia Animal e Patologia. Laboratório de Biologia do Reconhecimento. Campos, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular Aplicada a Micobactérias. Rio de Janeiro, RJ, Brasil.St. Petersburg Pasteur Institute. Laboratory of Molecular Microbiology. St. Petersburg. Russian Federation.Universidade Estadual do Norte Fluminense.Laboratório de Morfologia Animal e Patologia. Laboratório de Biologia do Reconhecimento. Campos, RJ, Brasil.Strains of the Beijing genotype family of Mycobacterium tuberculosis are a cause of particular concern because of their increasing
dissemination in the world and their association with drug resistance. Phylogenetically, this family includes distinct ancient
and modern sublineages. The modern strains, contrary to the ancestral counterparts, demonstrated increasing prevalence in
many world regions that suggest an enhanced bacterial pathogenicity. We therefore evaluated virulence of modern versus ancient
Beijing strains with similar epidemiological and genotype characteristics. For this, we selected six strains that had very similar
24-locus mycobacterial interspersed repetitive-unit–variable-number tandem-repeat (MIRU-VNTR) typing profiles and belonged
to the region of difference 181 (RD181) subgroup but differed using markers (mutT2 and mutT4 genes and NTF locus)
that discriminate between modern and ancient Beijing sublineages. The strains were isolated from native patients in Brazil and
Mozambique, countries with a low prevalence of Beijing strains. The virulence levels of these strains were determined in models
of pulmonary infection in mice and in vitro macrophage infection and compared with that of a strain from Russia, part of the
epidemic and hypervirulent Beijing clone B0/W148, and of the laboratory strain H37Rv. The results showed that two of the three
modern Beijing strains were highly pathogenic, exhibiting levels of virulence comparable with that of the epidemic Russian
strain. In contrast, all isolates of the ancient sublineage displayed intermediate or low virulence. The data obtained demonstrate
that the strains of the modern Beijing sublineage are more likely to exhibit highly virulent phenotypes than ancient strains and
suggest that genetic alterations characteristic of the modern Beijing sublineage favor selection of highly virulent bacteria
Correction: Hypervirulent Mycobacterium tuberculosis strain triggers necrotic lung pathology associated with enhanced recruitment of neutrophils in resistant C57BL/6 mice.
[This corrects the article DOI: 10.1371/journal.pone.0173715.]
Hypervirulent <i>Mycobacterium tuberculosis</i> strain triggers necrotic lung pathology associated with enhanced recruitment of neutrophils in resistant C57BL/6 mice
<div><p>Tuberculosis (TB) is a chronic infectious disease caused by <i>Mycobacterium tuberculosis</i> (Mtb) that in most cases induces irreversible necrosis of lung tissue as a result of excessive inflammatory reactions. The murine model of TB in resistant C57BL/6 mice infected with reference Mtb strains is widely used in TB studies; however, these mice do not show a necrotic pathology, which restricts their use in studies of irreversible tissue damage. Recently, we demonstrated that necrotic lung lesions could be induced in the C57BL/6 mice by highly virulent Mtb strains belonging to the modern Beijing sublineage. However, the pathogenic mechanisms leading to necrosis in this model were not elucidated. In this study, we investigated the dynamics of lung lesions in mice infected with highly virulent Beijing Mtb strain M299, compared with those infected with laboratory Mtb strain H37Rv. The data demonstrate that necrotic lung lesions in mice infected by the strain M299 were associated with enhanced recruitment of myeloid cells, especially neutrophils, and increased levels of proinflammatory cytokines, consistent with exacerbated inflammation. High levels of IFN-γ production contributed to the control of bacterial growth. Further progression to chronic disease was associated with a reduction in the levels of inflammatory mediators in the lungs, the accumulation of foamy macrophages and partial healing of the necrotic tissue by fibrosis. At a late stage of disease, degradation of foamy cells resulted in the liberation of accumulated lipids and persisting bacilli and further activation of inflammation, which promoted lung consolidation. Overall, our studies show that C57BL/6 mice infected with highly virulent Mtb strain may serve as a TB model reproducing an exacerbated inflammatory response in a resistant host to hypervirulent mycobacteria, leading to irreversible necrotic lung lesions.</p></div
Lung histopathology at the early chronic stage of infection by highly virulent Mtb strain M299.
<p>Pathological alterations to lung tissue were observed on day 120 after infection by microscopic analysis of lung sections stained by hematoxylin-eosin (<b>A</b> and <b>B</b>), Masson´s trichrome (<b>C</b>) and Ziehl-Neelsen (<b>D</b>). Panel <b>A</b> demonstrates a necrotic lesion (black star) partially healed by fibrosis and surrounded by a rim of lymphocytes and lipid-laden foamy macrophages in the external region. Secondary granulomatous lesions are marked by white stars. Panel <b>B</b> demonstrates a higher-magnification image of two compact perivascular lymphocyte granulomas, surrounded by foamy macrophages. <b>C</b>. Masson’s staining revealed the presence of profuse collagen (in light-blue) in the central area of the necrotic lesion. Note cellular debris in the bronchiole and the absence of a bronchiolar epithelium (black arrow). <b>D</b>. Single AFB, exhibiting weak staining (black arrow heads), could be seen in necrotic areas partially healed by fibrosis and in foamy macrophages within alveoli. Bars represent 500 μm- in the panel <b>A</b>, 200 μm- in the panels <b>B</b> and <b>C</b>; and 100 μm- in the panel <b>D</b>.</p
Kinetics of myeloid cell recruitment into the lungs of mice infected with strains M299 and H37Rv.
<p><b>A</b>. Total numbers of cells recovered from the lungs over a follow-up period of 60 days after inoculation with bacilli or sterile PBS (control, day 0). Proportions of viable and dead cells were determined by trypan-blue staining. <b>B</b>. Myeloid cell populations were identified by flow cytometry. Total numbers of CD11b<sup>+</sup> myeloid cells per lung and individual myeloid cell subpopulations, including neutrophils (CD11b<sup>+</sup>Ly6G<sup>+</sup>Ly6C<sup>+</sup>cells), immature neutrophil precursors (CD11b<sup>+</sup>Ly6G<sup>low</sup>Ly6C<sup>dim</sup>), inflammatory monocytes and macrophages (CD11b<sup>+</sup>Ly6C<sup>hi</sup>CD11c<sup>-</sup>Ly6G<sup>-</sup> cells), monocyte-derived dendritic cells (CD11b<sup>+</sup>CD11c<sup>+</sup>Ly6C<sup>hi</sup>Ly6G<sup>-</sup>) and dendritic cells (CD11b<sup>+</sup>CD11c<sup>+</sup>Ly6C<sup>-</sup>Ly6G<sup>-</sup>), were calculated. Values shown are the mean numbers of cells per lung ± SD, n = 8–9 lungs at each time point, pooled data from three independent experiments. Mean values of groups infected by strain M299 that were significantly different from the respective mean value of groups infected by H37Rv strain are indicated by asterisks * (p < 0.05). Significant differences between groups infected by each strain are indicated by lines and octothorpes #, (p < 0.05).</p
Kinetics of bacterial growth in the lungs and morbidity of C57BL/6 mice inoculated with Mtb strains.
<p>Mice were infected i.t. with 100 CFU of the hypervirulent strain M299 and laboratory strain H37Rv. <b>A.</b> Bacterial burdens in the lungs were quantified by CFU counting on days 0, 5, 21, 28, 120 and 150 after infection. Data from three experiments (n = 3 for each group and each experiment) are expressed logarithmically as the mean log<sub>10</sub> CFU standard deviation (SD) (error bars). Mean values that were significantly different from the respective mean value of the group infected by strain H37Rv are indicated by asterisks *, p < 0.05. <b>B</b>. Kinetics of post-infection changes in mouse body weight. Weight loss was used as an indicator of morbidity. Data are presented as the percentage of peak body weight of each mice (n = 20, pooled data from two independent experiments). Statistical differences are indicated by asterisks *, p < 0.05.</p
Kinetics of lung histopathology during the acute stage of infection in C57BL/6 mice inoculated with strain M299.
<p>Mice were intratracheally infected with ~100 CFU. Pathologic alterations of lung tissue were determined on day 15 p.i. (<b>A</b>), day 21 p.i. (<b>B, C</b>) and day 28 p.i. (<b>D- I</b>) by microscopy of lung sections stained with hematoxylin-eosin (<b>A- F</b>) and Ziehl-Neelsen (<b>G- I</b>). Panel <b>A</b> shows interstitial granulomatous infiltrates on day 15 p.i. Panels <b>B</b> and <b>C</b> show the development of alveolitis through infiltration of alveoli (black arrows) with inflammatory cells, predominantly by neutrophils (enclosed in black circles). Panels <b>D</b> and <b>G</b> show coalescing foci of tuberculous pneumonia leading to the formation of tubercules with extensive areas of central caseous necrosis (black stars). The peripheral zone of the necrotic lesion is amplified in <b>E</b> and <b>H</b>, demonstrating areas of alveolitis (black arrows) with thickened alveolar walls and intra-alveolar cellular exudates composed predominantly of neutrophil debris (<b>E</b>) and numerous AFB (<b>H</b>). The caseous core is amplified in <b>F</b> and <b>I</b>. Note the thrombosed septal capillaries (black arrow heads) and obstruction of small airways with cellular debris (white arrow) in the peripheral rim of the necrotic zone (<b>F</b>). Numerous extracellular AFB and clumps of bacteria can be seen in the necrotic area (<b>I</b>). Data are representative of four independent experiments. Bars represent 500 μm—in the panels D and G, 200 μm—in B, E and H, 100 μm—in A, F and I and 20 μm—in C.</p