Detection of proteases from Sporosarcina aquimarina and Algoriphagus antarcticus isolated from Antarctic soil.

Two psychrophilic bacterial samples were isolated from King George Island soil, in Antarctica. The phylogenetic analysis based on the 16S rRNA (rrs) gene led to the correlation with the closest related isolates as Sporosarcina aquimarina (99%) and Algoriphagus antarcticus (99%), with query coverage of 99% and 98%, respectively. The spent culture media from both isolates displayed proteolytic activities detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis containing gelatin as protein substrate. Under the employed conditions, S. aquimarina showed a 55 kDa protease with the best activity detected at pH 7.0 and at 27°C. A. antarcticus also showed a single extracellular protease, however its molecular mass was around 90kDa and its best activity was detected at pH 9.0 and at 37°C. The proteases from both isolates were inhibited by 1,10-phenanthroline and EDTA, two metalloprotease inhibitors. This is the first record of protease detection in both species, and our results may contribute to broaden the basic knowledge of proteases from the Antarctica environment and may help prospecting future biotechnological applications of these enzymes.

Serine protease activities in Oxysarcodexia thornax (Walker) (Diptera: Sarcophagidae) first instar larva.

We report for the first time the expression of multiple protease activities in the first instar larva (L1) of the flesh fly Oxysarcodexia thornax (Walker). Zymographic analysis of homogenates from freshly obtained L1 revealed a complex proteolytic profile ranging from 21.5 to 136 kDa. Although some activities were detected at pH 3.5 and 5.5, the optimum pH for most of the proteolytic activities was between pH 7.5 and 9.5. Seven of 10 proteases were completely inactivated by phenyl-methyl sulfonyl-fluoride, suggesting that main proteases expressed by L1 belong to serine proteases class. Complete inactivation of all enzymatic activities was obtained using N-p-Tosyl-L-phenylalanine chloromethyl ketone (100 microM), a specific inhibitor of chymotrypsin-like serine proteases.

Serine protease activities in Oxysarcodexia thornax (Walker) (Diptera: Sarcophagidae) first instar larva

We report for the first time the expression of multiple protease activities in the first instar larva (L1) of the flesh fly Oxysarcodexia thornax (Walker). Zymographic analysis of homogenates from freshly obtained L1 revealed a complex proteolytic profile ranging from 21.5 to 136 kDa. Although some activities were detected at pH 3.5 and 5.5, the optimum pH for most of the proteolytic activities was between pH 7.5 and 9.5. Seven of 10 proteases were completely inactivated by phenyl-methyl sulfonyl-fluoride, suggesting that main proteases expressed by L1 belong to serine proteases class. Complete inactivation of all enzymatic activities was obtained using N-p-Tosyl-L-phenylalanine chloromethyl ketone (100 µM), a specific inhibitor of chymotrypsin-like serine proteases.

[The effect of different strains Babesia bovis (Babes, 1888) on tick of Boophilus microplus (Canestrini, 1887)]. / Efeito de diferentes amostras de Babesia bovis (Babes, 1888) em carrapatos Boophilus microplus (Canestrini, 1887).

Two samples (modified and no-modified) of Babesia bovis, were used to evaluate the effect on engorged females of Boophilus microplus Babesia spp - free. For so much, were used three holstein breed bovine, males with 6 months of age, (BH1, BH2 and BH3), coming of the Plateau of Itatiaia, Rio de Janeiro, Brazil. Each calf was infested with 0.2g of B. microplus larvae Babesia spp- free, for 10 consecutive days. They were inoculated in the calves BH1 and BH2, modified and no modified sample, respectively. After natural fall of adults ticks from the vertebrate host, 100 engorged females of each host (BH1, BH2 and BH3) were incubated in BOD, with superior Relative Humidity up 80%, Temperature of 28 degrees C, were appraised daily until the 12th day post incubation (dpi.). In the group of the engorged females obtained of calf BH1, infection of ticks was not observed, to the haemolymph exam and oviposture. The sporokinets absence in these samples was related with the successive passages in splenectomized calves and the criopreservation of the sample. In engorged females from BH2 it was observed rates of infection of 100% and 82% of mortality in 7th dpi. and 100% until 12nd dpi., as the oviposture, 70% did not make oviposition and 30% made it until 7th dpi.

Dinâmica da infecção de Babesia bovis (Babés, 1888, Starcovici, 1893) em fêmeas ingurgitadas e ovos de Boophilus microplus (Canestrini, 1887) / Dynamic of infection of Babesia bovis (Babés, 1888, Starcovici, 1893) in engorged females and eggs of Boophilus microplus (Canestrini, 1887)

A dinâmica da infecção de B. bovis no carrapato-vetor B. microplus foi estudada em condições laboratoriais na Universidade Federal Rural do Rio de Janeiro no Laboratório de Protozoologia. Para tanto, foram examinadas 100 fêmeas ingurgitadas que se desprenderam naturalmente do hospedeiro vertebrado, sendo que 84 fêmeas apresentaram-se infectadas com esporocinetos de B. bovis, com a seguinte distribuição: 39 por cento, 33 por cento, e 12 por cento nos dias 3, 4 e 5 de incubação, respectivamente. Foram obtidas amostras de ovos provenientes das fêmeas positivas para B. bovis, 100 por cento das amostras de ovos estavam infectadas, apresentado a seguinte distribuição: 46,4 por cento, 34,5 por cento, 16,7 por cento e 2,4 por cento nos dias 4, 5, 6 e 7 de incubação, respectivamente. As freqüências acumuladas, tanto de fêmeas infectadas (84 por cento) quanto de ovos infectados (100 por cento) mantiveram-se até o 17° dia de incubação. De acordo com as freqüências acumuladas e o aumento do grau de infecção, conclui-se que amostras coletadas a partir do 5° e 7° dia de incubação, são ideais para o diagnóstico de B. bovis, em hemolinfa e ovos, respectivamente.

[Morphologic characterization and biological aspects of evolutive forms of Babesia bigemina (Smith and Kilborne, 1893) (Protozoa: Babesiidae) in Boophilus microplus (Canestrini, 1887)]. / Caracterizçã morfológica e aspectos biológicos das formas evolutivas de Babesia bigemina (Smith; Kilborne, 1893) (Protozoa: Babesiidae) em Boophilus microplus (Canestrini, 1887).

The development of Babesia bigemina in Boophilus microplus were studied in experimental conditions, using crossed-breed bovine from free-area of these parasites. Stages of the hemoparasites were observed in the tick vector, starting from the infected red-blood cells observed in the gut of engorged females, from the first 24 hours after detachment to the emergence of sporokinets in the larvas. In the period from 24 to 48 hours after detachment of the engorged females (DEF), the presence of some infected red-blood cells was verified, beside the occurrence of ray bodies and of vermiculars forms, known as oocynets. Since 72 hours after the DEF, the of okinets presence was observed in the cytoplasm of the epithelium cells besides of great sporokinets number in development. At same period, the presence of sporokinets of B. bigemina in the hemolymph samples was observed inside the hemocytes. After the fourth day of incubation beside the presence of the sporokinets was also verified in the Malpighi's tubes and ovaries. As well as in the ticks eggs from the sporokinets were also observed ticks eggs from the fourty day after the natural detachment of the engorged females of the host.

Distúrbios osteomusculares em uma comunidade da periferia de Montes Claros - MG: uma proposta de intervenção

Este estudo teve como objetivo elaborar um plano de ação para diminuir a prevalência e incidência de distúrbios osteomusculares relacionados ao trabalho entre os usuários da ESF Independência II. No diagnóstico situacional observou-se um elevado número de pessoas portadoras de tais distúrbios. Baseando-se neste problema foram selecionados os seguintes nós críticos: falta de conhecimento da população sobre exercícios físicos, alongamentos e correta ergonomia e falta de profissionais para orientar sobre ergonomia e exercícios físicos. Baseado nesses nós críticos foram propostas as seguintes ações de enfrentamento: criação do projeto "Educando" para explicação sobre a importância dos exercícios físicos e correta ergonomia e do projeto "Menos Dor" que terá a participação de acadêmicos de fisioterapia e educação física para orientar os pacientes no dia do "Grupo da Dor do Trabalho" sobre a forma correta de execução dos exercícios