Preliminary data of a project about the ecology of the Phlebotominae (sandflies) in a tegumentary leishmaniasis area of coconut plantation in the south of the State of Bahia, Brazil are presented. There are 60 dwellings far of one another, where existed 31 dogs and 229 inhabitants. Among them 41.5% were Montenegro positive; 37.5% from these had scars of healed ulcers and 8.8% had active ulcers. 53% of the house had dogs from which 22% were seropositive; from the 7 dogs with ulcers only 3 were seropositive. 14% of the inspected house harboured sandflies inside them. During two years of observations 72 sentinels hamsters were maintened at houses of patients with leishmaniasis ulcers, but they did not get the infection. During two years of observation, monthly collectings of sandflies were made inside house, chicken pen, curral, tree trunks and open field around a house with a patient with leishmaniasis. The following results were already obtained about the vectors: 5,614 specimens were identified as belonging to fourteen different species of sandflies. Among them Lutzomyia whitmani (92%) and Lutzomyia intermedia (4.8%) were the most abundant species. They are very anthropophilic and can be found inside houses and at peridomestic sites. Probably they are the principal vectors of the disease at domestic places. The other twelve species were less frequent and more found at sylvatic places in inspite they also bite the man. Generally the biting activity of most of the species of the area begins at 5 p.m. in the dusk and reaches its peak at 0 hour a.m., when begins declining until disapear at 7 a.m. L. whitmani was similarly collected with the same density in all lunar phases while L. intermedia was more abundant during the new moon phase. Most of the hundreds sandflies collected during the second year of observations, remains preserved in liquid nitrogen, watching for the adjustment of PCR molecular techniques to be processed for determination of the vector natural infection rates with leishmanias. Final results on all the project will be published as soon as the examination of such material has been processed.
The major components of protein extracts from the cattle tick Boophilus microplus eggs and larvae of various ages were characterized by molecular sieving chromatography, ion exchange chromatography and SDS-PAGE. The fractions analysed showed a changing chromatographic pattern development. A serum raised against the components of a fraction showing characteristics of vitellin strongly reacted in Western blots with the major peptides of extracts from eggs, larvae, gut and ovary. Comparison of patterns obtained by electrophoresis in non-denaturing PAGE, stained with Coomassie blue or with benzidine/hydrogen peroxide, revealed that the major proteins of these extracts are haemoproteins, possibly in different aggregation states or heterogeneous in composition.
Bacillus anthracis was isolated and identified from a bacterial collection of samples from the Amazon river bank. Type II restriction endonuclease activity was detected in this prokaryote, the enzyme was purified, the molecular mass of the native protein estimated by gel filtration, and optima pH, temperature and salt requirements were determined. Quality control assays showed complete absence of 'non-specific nucleases'. Restriction cleavage analysis and DNA sequencing of restriction fragments allowed unequivocal demonstration of 5'-GG downward arrow CC-3' as the recognition sequence. This enzyme was named BanAI and is therefore an isoschizomer of the prototype restriction endonuclease HaeIII. This is the first report of a type II restriction endonuclease identified, purified from a natural isolate of B. anthracis.
A strain of Bacillus pumilus was isolated and identified from water samples collected from a small affluent of the Amazon River. Type II restriction endonuclease activity was detected in these bacteria. The enzyme was purified and the molecular weight of the native protein estimated by gel filtration and SDS-PAGE. The optimum pH, temperature and salt requirements were determined. Quality control assays showed the complete absence of "nonspecific nucleases." Restriction cleavage analysis and DNA sequencing of restriction fragments allowed the unequivocal demonstration of 5´GAG↓CTC3´ as the recognition sequence. This enzyme was named BpuAmI and is apparently a neoschizomer of the prototype restriction endonuclease SacI. This is the first report of an isoschizomer and/or neoschizomer of the prototype SacI identified in the genus Bacillus.
The restriction endonuclease BliAI, an isoschizomer of ClaI, which recognizes the sequence 5´-AT↓CGAT -3´, was purified from a natural isolate identified as Bacillus licheniformis. The restriction endonuclease was isolated from cell extracts using single-step purification by phosphocellulose column chromatography. The restriction endonuclease is active at 37ºC and over a wide range of pH and salt concentration. The molecular weight of the purified restriction enzyme is consistent with a value of 39 kDa.
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