RESUMO
Leptospirosis is a zoonotic disease worldwide and caused by the pathogenic spirochetes of the genus Leptospira. Bacterins make up the vaccines used against leptospirosis, but they only succeed in providing short-term and serovar-specific protection. The use of Mycobacterium bovis BCG as a live vaccine vector expressing leptospiral antigens is a promising alternative, particularly due to its adjuvant properties. Four distinct portions P1 (lipL32), P2 (ligAni), P3 (lemA:ligAni) and P4 (lipL32:lemA) of a recombinant chimera composed of the lipL32, lemA and ligANI genes from Leptospira interrogans were cloned individually according to the BioBricks® strategy in the plasmid pUP500/PpAN. These constructs were individually transformed into a BCG Pasteur strain, and protein expression was detected by Western blot. For vaccination, 5 groups of 10 Golden Syrian hamsters were used, aged 4-6 weeks - group 1, rBCG (LipL32); group 2, rBCG (LigAni); group 3, rBCG (LemA:LigAni); group 4, (LipL32:LemA); group 5, wild-type BCG Pasteur (negative control). Two doses containing 106 CFU of rBCG were administered subcutaneously, the challenge was performed with 5 × LD50 of Leptospira interrogans serovar Copenhageni L1-130, and the animals were observed for a 30-day period until the endpoint was reached. Humoral immunity was assessed via indirect ELISA, while renal colonisation was assessed by culture and quantitative real-time PCR. All vaccinated groups were protected against lethal challenge and renal colonisation, in comparison with negative control group (P < 0.05). Recombinant vaccines were not effective at inducing significant humoral immunity, which suggests the induction of cellular immunity - a characteristic of M. bovis BCG. In conclusion, all formulations provide 100% significant protection against leptospirosis in hamsters with no renal colonisation. The use of rBCG as a vaccine vector represents a promising alternative for the control of animal leptospirosis, allowing for protection against clinical signs of leptospirosis and renal colonisation.
Assuntos
Leptospira interrogans , Leptospira , Leptospirose , Mycobacterium bovis , Animais , Antígenos de Bactérias/genética , Vacina BCG , Vacinas Bacterianas , Cricetinae , Leptospira interrogans/genética , Leptospirose/prevenção & controle , Mycobacterium bovis/genéticaRESUMO
Leptospirosis is a zoonosis of worldwide distribution, caused by infection with pathogenic Leptospira species. The vaccines that are currently available are bacterins, with limited human use, that confer short-term, serovar-specific immunity. Lig proteins are considered to be the best vaccine candidates to date. Here, we aimed to construct a recombinant Lig chimera (LC) comprised of LigAni and LigBrep fragments, and to evaluate it as subunit or DNA vaccine using different administration strategies. Vaccines were formulated with 50⯵g of recombinant LC associated with different adjuvants or with 100⯵g of pTARGET/LC. Four-week-old hamsters received two doses of vaccine with different strategies and were challenged with 5â¯×â¯DL50Leptospira interrogans serovar Copenhageni strain Fiocruz L1-130. The immune response generated by Lig chimera conferred 100% protection to hamsters treated with at least one dose of recombinant LC. Despite the high levels of antibodies that vaccinated animals produced, a sterilizing immunity was not achieved. The lack of a sterilizing immunity could indicate the importance of a mixed humoral and cellular immune response. The present study generated insights that will be useful in the future development of improved subunit vaccines against leptospirosis.
Assuntos
Antígenos de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Leptospira/imunologia , Leptospirose/prevenção & controle , Proteínas Recombinantes de Fusão/imunologia , Vacinas de Subunidades Antigênicas/imunologia , Adjuvantes Imunológicos , Animais , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/genética , Vacinas Bacterianas/genética , Células CHO , Cricetinae , Cricetulus , Expressão Gênica , Humanos , Leptospira/genética , Leptospirose/imunologia , Leptospirose/microbiologia , Leptospirose/mortalidade , Proteínas Recombinantes de Fusão/genética , Vacinas de DNA , Vacinas de Subunidades Antigênicas/genéticaRESUMO
As ações municipais no campo do esporte e do lazer são multifacetadas. O objetivo deste tex-to é verificar as ações de esporte e lazer da Prefeitura Municipal de Pelotas - RS, na gestão 2009-2012. Recolhemos dados de diversas secretarias que, de alguma forma, atuam no campo do esporte e do lazer, bem como dados do Plano Diretor do município de Pelotas - RS. Entre-vistas com envolvidos e observação dos espaços públicos de práticas de esportes e lazer tam-bém foram realizadas. Constatamos que existem alguns programas que não dão conta das ne-cessidades da população, além da precariedade dos equipamentos públicos, como praças e campos esportivos. Isso traz grandes barreiras para a população pelotense ao direito ao espor-te e ao lazer.
The municipal actions in the field of sports and leisure are multifaceted. The aim of this paper is to check the sports and leisure activities of the City of Pelotas, management 2009-2012. To this end, data from several departments that somehow work in the field of sports and leisure were collected, as well as the data from the Master Plan of Pelotas / RS. Involved agents were interviewed with we observed public spaces for sports and recreational spaces. We conclude that there are some programs that do not realize the population's needs, as well as we verified the precariousness of public spaces such as parks and sports fields. This brings great barriers for Pelotas population in relation to the right to sport and leisure.
Las actuaciones municipales en el ámbito de los deportes y del ocio son diversas. El objetivo de este trabajo es comprobar las actividades deportivas y de ocio del Ayuntamiento de la ciu-dad de Pelotas, gestión 2009-2012. Con este fin, se recogieron datos de varios departamentos que trabajan de alguna manera en el campo del deporte y el ocio, así como datos del Plan Di-rector del municipio de Pelotas RS. También se realizaron entrevistas con personas vincula-das a ello y fueron observados los espacios públicos para la práctica del deporte y del ocio. Llegamos a la conclusión de que hay algunos programas que no satisfacen las necesidades de la población, así como que son precarias las instalaciones públicas, como plazas y campos deportivos. Esto conlleva grandes barreras para que la población Pelotas ejerza el derecho al deporte y el ocio.