New Strategies on Molecular Biology Applied to Microbial Systematics

Systematics is the study of diversity of the organisms and their relationships comprising classification, nomenclature and identification. The term classification or taxonomy means the arrangement of the organisms in groups (rate) and the nomenclature is the attribution of correct international scientific names to organisms and identification is the inclusion of unknown strains in groups derived from classification. Therefore, classification for a stable nomenclature and a perfect identification are required previously. The beginning of the new bacterial systematics era can be remembered by the introduction and application of new taxonomic concepts and techniques, from the 50’s and 60’s. Important progress were achieved using numerical taxonomy and molecular taxonomy. Molecular taxonomy, brought into effect after the emergence of the Molecular Biology resources, provided knowledge that comprises systematics of bacteria, in which occurs great evolutionary interest, or where is observed the necessity of eliminating any environmental interference. When you study the composition and disposition of nucleotides in certain portions of the genetic material, you study searching their genome, much less susceptible to environmental alterations than proteins, codified based on it. In the molecular taxonomy, you can research both DNA and RNA, and the main techniques that have been used in the systematics comprise the build of restriction maps, DNA-DNA hybridization, DNA-RNA hybridization, sequencing of DNA sequencing of sub-units 16S and 23S of rRNA, RAPD, RFLP, PFGE etc. Techniques such as base sequencing, though they are extremely sensible and greatly precise, are relatively onerous and impracticable to the great majority of the bacterial taxonomy laboratories. Several specialized techniques have been applied to taxonomic studies of microorganisms. In the last years, these have included preliminary electrophoretic analysis of soluble proteins and isoenzymes, and subsequently determination of deoxyribonucleic acid base composition and assessment of base sequence homology by means of DNA-RNA hybrid experiments beside others. These various techniques, as expected, have generally indicated a lack of taxonomic information in microbial systematics. There are numberless techniques and methodologies that make bacteria identification and classification study possible, part of them described here, allowing establish different degrees of subspecific and interspecific similarity through phenetic-genetic polymorphism analysis. However, was pointed out the necessity of using more than one technique for better establish similarity degrees within microorganisms. Obtaining data resulting from application of a sole technique isolatedly may not provide significant information from Bacterial Systematics viewpoint

Hybrid systems biology: application to Escherichia coli

Dissertation presented to obtain a Master degree in Biotechnology

Development of paper-based color test-strip for drug detection in aquatic environment: Application to oxytetracycline

The wide use of antibiotics in aquaculture has led to the emergence of resistant microbial species. It should be avoided/minimized by controlling the amount of drug employed in fish farming. For this purpose, the present work proposes test-strip papers aiming at the detection/semi-quantitative determination of organic drugs by visual comparison of color changes, in a similar analytical procedure to that of pH monitoring by universal pH paper. This is done by establishing suitable chemical changes upon cellulose, attributing the paper the ability to react with the organic drug and to produce a color change. Quantitative data is also enabled by taking a picture and applying a suitable mathematical treatment to the color coordinates given by the HSL system used by windows. As proof of concept, this approach was applied to oxytetracycline (OXY), one of the antibiotics frequently used in aquaculture. A bottom-up modification of paper was established, starting by the reaction of the glucose moieties on the paper with 3-triethoxysilylpropylamine (APTES). The so-formed amine layer allowed binding to a metal ion by coordination chemistry, while the metal ion reacted after with the drug to produce a colored compound. The most suitable metals to carry out such modification were selected by bulk studies, and the several stages of the paper modification were optimized to produce an intense color change against the concentration of the drug. The paper strips were applied to the analysis of spiked environmental water, allowing a quantitative determination for OXY concentrations as low as 30 ng/mL. In general, this work provided a simple, method to screen and discriminate tetracycline drugs, in aquaculture, being a promising tool for local, quick and cheap monitoring of drugs.

A systems biology framework for pathway level culture media engineering: pplication to Pichia pastoris cultures

Dissertação para obtenção do Grau de Doutor em Engenharia Química e Bioquímica

Biology of Triatominae (Reduviidae Hemiptera) from North of Formosa County (Goiás-Brazil) I. Length of life cycle of Triatoma sordida (Stal. 1859)

In the present paper the life cycle of Triatoma sordida was studied. The mean length from egg to adult was 213 days. The mean length in days from each stage was: 24.3 (± 1.30) for the first. 32.8 (± 1.45) (2nd), 36.1 (± 1.50) (3rd), 44.6 (± 1.85) (4th) and 52.0 (± 1.92) (5th). The mean egg incubation períod was 23.2 (± 1.40). Overall mortality was 18.8% and egg viability was 82.5%.

Biology of Triatominae (Reduviidae, Hemiptera) from north of formosa county (Goiás - Brazil) II. Length of life cycle of Rhodnius Neglectus lent, 1954

In the present paper the life cycle of Rhodnius neglectus was studied. The mean length from egg to adult was 119 days. The mean length in day from each stage was: 17.4 (± 1.15) for first, 18.2 (± 1.28) for second, fourth and 29.8 (± 1.46) for fifth.

Biology of triatominae (Reduviidade, Hemiptera) from north of Formosa county (Goiás - Brazil). III: length of life cycle of Psammolestes tertius Lent and Jurberg 1965

In the present work the life cycle of Psammolestes tertius was studied. The mean length, in days, fromeach stage was: 26.3 (± 1.7) (1st), 28.6 (± 1.8) (2nd), 28.4 (± 1.8) (3rd), 32.2 (± 1.9) (4th) and 33.5 (± 5.8) (5th). The mean egg incubation period was 15.7 days (± 1.7). Overall mortality was 48.9% and egg viability was 65.7%.

Establishing a cell biology platform: isolation and preservation of human blood products

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

FSR Quorum Sensing: Role in Enterococcus faecalis Biology & Host Infection

Dissertation presented to obtain the Ph.D degree in Biology.