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Molecular sequence data together with ultrastructural features were used to infer the phylogenetic position of Tuberculina species. Additional ultrastructural characteristics were used to determine their mode of nutrition. We investigated ultrastructural morphology of the type species Tuberculina persicina and determined base sequences from the D1/D2 region of the nuclear large-subunit ribosomal DNA of the three commonly distinguished Tuberculina species, T. maxima, T. persicina and T. sbrozzii. Analyses of sequence data by means of a Bayesian method of phylogenetic inference using a Markov Chain Monte Carlo technique reveal the basidiomycetous nature of Tuberculina. Within the Urediniomycetes, Tuberculina clusters as a sister group of Helicobasidium, closely related to the rusts (Uredinales). This phylogenetic position is supported by the uredinalean architecture of septal pores in Tuberculina. In addition, we present aspects of the ultrastructural morphology of the cellular interaction of Tuberculina and rusts showing a unique interaction with large fusion pores, revealing the mycoparasitic nature of Tuberculina on its close relatives, the rusts.

In the study, we found rust in pistachio fruits and verified that it was caused by Tuberculina persicina , anamorph of Helicobasidium purpureum , through isolation and culture, microscopic observation, molecular identification and pathogenicity tests of Koch’s postulates. To our knowledge, this is the first report of rust fruit in pistachio caused by Tuberculina persicina .

INTRODUÇÃO: A contagem de células CD4+ representa marcador da resposta imune celular em pacientes infectados pelo HIV-1. Testes cutâneos de hipersensibilidade tardia (DTH) podem ser empregados para avaliar in vivo respostas celulares a antígenos comuns. MÉTODOS: DTH para derivado proteico purificado de tuberculina (PPD), esporotriquina, tricofitina, candidina e estreptoquinase/estreptodornase foram realizados. Foram testados crianças/adolescentes infectados pelo HIV-1 (n=36) e indivíduos saudáveis (n=56), soronegativos para HIV-1/HIV-2 pareados por sexo-idade, todos com cicatriz vacinal por BCG. Teste exato de Fisher foi aplicado (p<0,05). RESULTADOS: Entre as crianças/adolescentes infectados pelo HIV-1, mediana de idade=8,1 anos; 20/36 eram do sexo masculino; 35 casos de transmissão vertical; 34 casos de AIDS sob terapia antirretroviral; mediana de carga viral = 3.04lc10 cópias/ml; mediana de contagem de células CD4+ = 701 células/μl. Entre os infectados e saudáveis a reatividade DTH a pelo menos um dos antígenos foi, respectivamente, 25% (9/36) e 87,5% (49/56) (p<0,001). Reatividade à candidina predominou nos infectados (8/36, 22%) e ao PPD nos indivíduos saudáveis (40/56, 71,4%). A reatividade ao PPD entre infectados foi de 8,3% (p<0,01). A mediana da induração ao PPD foi 2,5mm (variação: 2-5mm) entre infectados e 6,0mm (variação: 3-15mm) entre os saudáveis. Não observamos correlação entre positividade ao PPD e idade. No grupo de infectados, não observamos correlação entre contagens de células CD4+ e reatividade ao DTH. CONCLUSÕES: Respostas DTH significativamente diminuídas, incluindo a reatividade ao PPD foram observadas em crianças/adolescentes infectados pelo HIV-1 comparadas com controles saudáveis, provavelmente refletindo doença avançada e supressão da imunidade mediada por células T.INTRODUCTION: Among HIV-1-infected patients, CD4+ T cell counts are well-established markers of cell-mediated immunity. Delayed-type hypersensitivity (DTH) skin tests can be used to evaluate in vivo cell-mediated immunity to common antigens. METHODS: DTH responses to tuberculin purified protein derivative (PPD), sporotrichin, trichophytin, candidin and streptokinase/streptodornase antigens were assessed. Thirty-six HIV-1-infected children/adolescents and 56 age- and sex-matched HIV-1/HIV-2-seronegative participants were tested. All participants had a BCG scar. Fisher's exact test was used to evaluate significant differences between groups (p<0.05). RESULTS: The main characteristics of the HIV-1 patients were as follows: median age 8.1 years; 20/36 were males; 35 were vertical transmission cases; 34 were AIDS cases under antiretroviral therapy; median viral load = 3.04 log10 copies/ml; median CD4+ T cell count = 701 cells/μl. A total of 25% (9/36) and 87.5% (49/56) of HIV-1-infected and healthy participants, respectively, displayed DTH reactivity to at least one antigen (p<0.001). Among HIV-1-infected participants, reactivity to candidin predominated (8/36, 22.2%), while PPD positivity prevailed among healthy participants (40/56, 71.4%). PPD reactivity in the HIV-1-positive group was 8.3% (p<0.01). The median PPD induration was 2.5mm (range: 2-5mm) in the HIV-1 group and 6.0 mm among healthy participants (range: 3-15mm). There was no correlation between PPD positivity and age. No correlation between CD4+ T cell counts and DTH reactivity was observed among HIV-1-infected patients. CONCLUSIONS: DTH skin test responses, including PPD reactivity, were significantly lower among HIV-1-infected participants compared to healthy controls, which likely reflects advanced disease and T cell depletion.

Tuberculina species are mitosporic parasites of rust fungi. It was demonstrated recently that Tuberculina represents the asexual life stage of the plant-parasitic genus Helicobasidium. Here we reveal the host specificities of Tuberculina and Helicobasidium species on rust fungal hosts by means of infection experiments and molecular analyses. We inoculated species of the rust genera Chrysomyxa, Coleosporium, Cronartium, Gymnosporangium, Puccinia, Tranzschelia and Uromyces with conidia and with basidiospores of Helicobasidium longisporum and H. purpureum and with conidia of Tuberculina maxima, T. persicina and T. sbrozzii. In addition we analyzed base sequences from the nuclear ITS region of 51 Tuberculina and Helicobasidium specimens collected in the field together with the sequences from the Tuberculina infections obtained by infection experiments. The resulting data show that at least six monophyletic lineages are within the Tuberculina/Helicobasidium- group that can be unambiguously distinguished by combining molecular and morphological characters and by specific host spectra of the Tuberculina-stage. This diversity opens up new vistas on the evolution of this exceptional mycoparasitic-phytoparasitic fungal group.

ABSTRACT Objective: to evaluate the risk of infection and illness caused by Mycobacterium tuberculosis among health care and security staff in prisons in two regions of Rio Grande do Sul (RS). Method: cross-sectional study involving prison staff. An interview and sputum smear microscopy and culture were performed. Latent infection was evaluated according to the result of the tuberculin test (TT), self-referred. Results: among staff who had a TT, 10 (83.3%) in the central region and 2 (16.7%) in the southern region were considered reactors. Length of employment among prison officers who reacted to TT was 15.3 years, and among health care workers, 4.1 years (p = 0.01). No cases of active tuberculosis (TB) were identified. Conclusion: prevalence of latent TB was 27.9%. Length of employment between different professional categories and their working regions was considered a risk factor for latent TB. RESUMEN Objetivo: evaluar el riesgo de infección y de la enfermedad por Mycobacterium tuberculosis entre los profesionales de la salud y seguridad en los centros penitenciarios en dos regiones del estado de Rio Grande do Sul (RS). Método: estudio transversal con la participación de profesionales de la prisión. Se llevó a cabo una entrevista, la baciloscopia y cultivo de esputo. La infección latente se evaluó de acuerdo con el resultado de la prueba de la tuberculina (TST) mediante auto-reporte. Resultados: entre los trabajadores que realizaron la TST en la región central, 10 (83,3%) fueron considerados reactores; y 2 (16,7%) en la región Sur. El tiempo de trabajo entre los agentes de la prisión con reacción a la TST fue de 15,3 años, y entre los trabajadores de la salud fue de 4,1 años (p = 0,01). No hubo casos identificados de la tuberculosis (TB) activa. Conclusión: la prevalencia de la TB latente fue del 27,9%. El tiempo de trabajo entre las diferentes categorías profesionales y la región en la que trabajan fueron considerados factores de riesgo para la TB latente. RESUMO Objetivo: avaliar o risco de infecção e adoecimento por Mycobacterium tuberculosis entre profissionais de saúde e de segurança em casas penitenciárias de duas regiões do Rio Grande do Sul (RS). Método: estudo transversal, envolvendo profissionais de penitenciárias. Foi realizada uma entrevista, baciloscopia e cultura de escarro. A infecção latente foi avaliada de acordo com o resultado do teste tuberculínico (TT), auto-referido. Resultados: entre os trabalhadores que realizaram o TT na região central, 10 (83,3%) foram considerados reatores; e na região sul, 2 (16,7%). O tempo de trabalho entre os agentes penitenciários reatores ao TT foi 15,3 anos e entre os trabalhadores da saúde 4,1 anos (p = 0,01). Não foram identificados casos de Tuberculose (TB) ativa. Conclusão: a prevalência de TB latente foi 27,9%. O tempo de trabalho entre as diferentes categorias profissionais e a região em que trabalham foram considerados fator de risco para TB latente.

The aim of this paper was to review the status of bovine tuberculosis (BTB) in Ethiopia in relation with the existing animal husbandry systems and abattoir meat inspection surveillances. Ethiopia is one of the African countries where tuberculosis is widespread in both humans and cattle. The disease is considered as one of the major livestock diseases that results in high morbidity and mortality, although the current status on the actual prevalence rate of BTB at a national level is yet unknown. Detection of BTB is based mainly on tuberculin skin testing, abattoir meat inspection and very rarely on bacteriological techniques. The prevalence rates vary from 3.4% (small farms) to 50% (intensive dairy farms) and from 3.5% to 5.2% in slaughterhouses. Control measures, economic impacts and zoonotic aspect of the disease are also briefly reviewed.

Objective: To estimate the prevalence of latent M. tuberculosis infection and identify characteristics related to latent infection among workers of the primary healthcare network. Methods: Cross-sectional, observational and descriptive study, conducted in 2011, included 137 basic health workers who performed their activities in a municipality. Interviews were carried out addressing characteristics of exposure at work (BCG vaccination, length of time working in the health system, potential exposure to tuberculosis, HIV infection, use of corticosteroids, diabetes, use of tobacco or alcohol) and subsequently performed tuberculin skin test (TST). Data was analyzed with Fischer and Student's t-test (p < 0.05). Results: Health professionals were mostly women (n=126, 92%) with mean age of 35.3 (± 9.1) years. Among the reactors, 25 (55.5%) sought medical care, and of these, 7 (28%) underwent prophylactic treatment. The prevalence of latent tuberculosis was 32.8%. Of the characteristics evaluated, only smoking (OR 3.03; 95%CI 1.05-8.77) was associated with latent infection. Conclusion: The estimated prevalence of latent M. tuberculosis infection among the evaluated health workers was 32.8%. As for the characteristics studied, it was not possible to relate them to latent infection among workers in primary healthcare network, with the exception of the smoking habit. Objetivo:Estimar a prevalência de infecção latente por M. tuberculosis e identificar características relacionadas à infecção latente entre os trabalhadores da rede básica de saúde. Métodos:Estudo transversal, observacional e descritivo, realizado em 2011, incluiu 137 trabalhadores da rede básica de saúde que desempenhavam suas atividades em um município. Realizaram-se entrevistas abordando características de exposição no trabalho (vacinação com BCG, tempo de trabalho na rede de saúde, exposição potencial à tuberculose, infecção pelo HIV, uso de corticoides, diabetes, uso de tabaco ou álcool) e, posteriormente, aplicouse o teste tuberculínico (TT). Analisaram-se os dados com teste Fischer e T-Student (p <0,05). Resultados:Os profissionais da saúde eram majoritariamente mulheres (n=126, 92%), com idade média de 35,3 (±9,1) anos. Entre os profissionais reatores, 25 (55,5%) buscaram assistência médica - destes, 7 (28%) realizaram tratamento profilático. A prevalência de tuberculose latente foi de 32,8%. Das características avaliadas, somente o tabagismo (OR:3,03; IC 95% 1,05-8,77) foi associado à infecção latente. Conclusão: A prevalência estimada de infecção latente por M tuberculosis entre os trabalhadores de saúde avaliados foi de 32,8%. Quanto às características estudadas, não foi possível relacioná-las com a infecção latente entre os trabalhadores da rede básica de saúde, com exceção do tabagismo. Objetivo:Estimar la prevalencia de infección latente por M. tuberculosis e identificar las características relacionadas a La infección latente de los trabajadores de la red básica de salud. Métodos: Estudio transversal, observacional y descriptivo realizado en 2011 con 137 trabajadores de la red básica de salud que desarrollaban sus actividades en un municipio. Se realizaron entrevistas sobre las características de exposición del trabajo (vacunación con BCG, tiempo de trabajo en la red de salud, exposición potencial a la tuberculosis, infección por el VIH, uso de corticoides, diabetes, uso de tabaco o alcohol) y, a posteriori, se aplico la prueba de la Tuberculina (PT). Los datos fueron analizados con la prueba de Fisher y el t-student (p < 0,05). Resultados:Los profesionales sanitarios eran en su mayoría mujeres (n=126, 92%) y edad media de 35,3 (±9,1) años. De los profesionales reatores, 25 (55,5%) buscaron la asistencia medica - de estos, 7 (28%) realizaron tratamiento profiláctico. La prevalencia de tuberculosis latente fue del 32,8%. De las características evaluadas solamente el tabaquismo (OR: 3,03; IC 95% 1,05-8,77) estuvo asociado a la infección latente.Conclusión:La prevalencia estimada de infección latente de M tuberculosis de los trabajadores sanitarios evaluados fue del 32,8%. No ha sido posible relacionar las características estudiadas con la infección latente entre los trabajadores de La red básica de salud, a excepción del tabaquismo.

The aim of this study was to evaluate the prevalence of bovine tuberculosis (TB) and the magnitude and trend of tuberculous lesions in slaughtered cattle (1994 to 2010) as well as tuberculin skin tests (TST) in 2,853 cattle (84 herds) of 39 livestock rearing communities in the highlands of Cameroon. Out of 129,165 slaughtered cattle inspected, 599 (0.46%; 95% CI: 0.43%-0.50%) showed suspected tuberculous lesions among a total of 983 (0.76%; 95% CI: 0.71%-0.81%) identified pathologies. The monthly TB detection rates ranged from 0.30% (95% CI: 0.20%-0.40%) to 0.81% (95% CI: 0.64%-0.98%) and annual rates from 0.04% (95% CI: 0%-0.11%) to 1.46% (95% CI: 1.22%-1.69%). The rates were not affected by season and fluctuating peaks were also recorded. The comparative TST revealed that bovine TB was widely distributed in live cattle (4.67%; 95% CI: 3.89%-5.44%) and was significantly higher in the Western highlands than Adamawa plateaux. Comparative TST bovine TB reactors were higher (P less than 0.05) in cattle managed in semi-intensive and beef production systems compared to the others. Animals in small herds showed signif. higher rates than those in large herds. Bovine TB prevalence in exotic/upgraded cattle was comparable to that of the Red Bororo zebu but signif. higher than the rates in Guadali and White Fulani zebus. Among the indigenous zebus, the rate was signif. higher in Red Bororo than the Guadali and White Fulani. Sex did not influence bovine TB prevalence in this study but diagnosis of the disease was signif. higher among adult/older cattle than in younger animals. Further analysis of the TST responses revealed that atypical mycobacterial infections was widespread and 6.83% of tested animals showed positive reactions at both bovine and avian tuberculin injection sites and a strong association between skin responses to both tuberculins. Our study confirms that bovine TB is prevalent in live cattle and abattoirs in Cameroon.

Cellular interaction of the basidiomycete Tuberculina persicina with the haploid stages of two rusts Puccinia silvatica and Tranzschelia pruni-spinosae was analyzed by serial-section electron microscopy of chemically fixed samples and samples subjected to high-pressure freezing and freeze substitution. Tuberculina persicina is a contact parasite, forming neither haustoria nor other intracellular structures. However, at contact areas between T. persicina and its hosts, distinct interfungal interactions are present. At the beginning, a hyphal cell of T. persicina invades the host cell wall with a protuberance and the cell walls of both protuberance and host cell dissolve at the point of contact. Thus, the plasma membranes of the two organisms contact and fuse to form a pore that enlarges to a final diameter of approximately 1 μm. The membrane of the fusion pore is continuous with the plasma membranes of both cells, and Tuberculina nuclei and other organelles are transferred to the rust cells. Phylogenetic and functional aspects of this curious basidiomycetous interfungal interaction are discussed, and a hypothesis of the evolutionary derivation of the Tuberculina mycoparasitism from a sexual interaction is presented.

Tuberculina species are mitosporic parasites of rust fungi. It was demonstrated recently that Tuberculina represents the asexual life stage of the plant-parasitic genus Helicobasidium. Here we reveal the host specificities of Tuberculina and Helicobasidium species on rust fungal hosts by means of infection experiments and molecular analyses. We inoculated species of the rust genera Chrysomyxa, Coleosporium, Cronartium, Gymnosporangium, Puccinia, Tranzschelia and Uromyces with conidia and with basidiospores of Helicobasidium longisporum and H. purpureum and with conidia of Tuberculina maxima, T. persicina and T. sbrozzii. In addition we analyzed base sequences from the nuclear ITS region of 51 Tuberculina and Helicobasidium specimens collected in the field together with the sequences from the Tuberculina infections obtained by infection experiments. The resulting data show that at least six monophyletic lineages are within the Tuberculina/Helicobasidium-group that can be unambiguously distinguished by combining molecular and morphological characters and by specific host spectra of the Tuberculina-stage. This diversity opens up new vistas on the evolution of this exceptional mycoparasitic-phytoparasitic fungal group.