A escolha da faixa espectral no uso combinado de métodos espectroscópicos e quimiométricos

A method is presented for the choice of spectral regions when absorption measurements are coupled to chemometric tools to perform quantitative analyses. The method is based on the spectral distribution of the relative standard deviation of concentration (s c/c). It has been applied to the development of PLS-FTNIR calibration models for the determination of density and MON of gasoline, and ethanol content and density of ethanol fuel. The new method was also compared with the correlation (R²) method and has proved to generate PLS calibration models that present better accuracy and precision than those based on R².

SISTEMAS ALTERNATIVOS DE CALIBRAÇÃO PARA DETERMINAÇÃO ESPECTROFOTOMÉTRICA SIMULTÂNEA DE ESPÉCIES COM INTERFERÊNCIA ESPECTRAL

Two simple and efficient procedures have been developed for the rapid simultaneous determination of compounds with mutual spectral interference (rifampicin (RIF) and isoniazid (INH)). The first method was based on the UV–Vis spectral signal (190–600 nm) of synthetic RIF and INH aqueous solutions, whereas the second method involved the visible spectral signal registered between 350 and 800 nm after the reaction of INH with a Cu2+/neocuproine complex. Both multivariate spectrophotometric methods show excellent prevision capacity, providing results that are statistically equivalent with those provided by the standard chromatographic procedure. The methods were validated according to criteria established by ANVISA, showing precision, accuracy and robustness compatible with the requirements for new analytical methods, additionally allowing the reduction of waste generation.

Sequences of the coat protein gene from brazilian isolates of Papaya ringspot virus

Papaya ringspot virus (PRSV) is the causal agent of the main papaya (Carica papaya) disease in the world. Brazil is currently the world's main papaya grower, responsible for about 40% of the worldwide production. Resistance to PRSV on transgenic plants expressing the PRSV coat protein (cp) gene was shown to be dependent on the sequence homology between the cp transgene expressed in the plant genome and the cp gene from the incoming virus, in an isolate-specific fashion. Therefore, knowledge of the degree of homology among the cp genes from distinct PRSV isolates which are present in a given area is important to guide the development of transgenic papaya for the control of PRSV in that area. The objective of the present study was to assess the degree of homology among the PRSV cp genes of several Brazilian isolates of this virus. Papaya and PRSV are present in many different ecosystems within Brazil. Twelve PRSV isolates, collected in eight different states from four different geographic regions, were used in this study. The sequences of the cp gene from these isolates were compared among themselves and to the gene used to generate transgenic papaya for Brazil. An average degree of homology of 97.3% at the nucleotide sequence was found among the Brazilian isolates. When compared to 27 isolates from outside Brazil in a homology tree, the Brazilian isolates were clustered with Australian, Hawaiian, and Central and North American isolates, with an average degree of homology of 90.7% among them.

Identification of Rhizoctonia solani associated with soybean in Brazil by rDNA-ITS sequences

The aim of this study was to identify isolates of Rhizoctonia solani causing hypocotyl rot and foliar blight in soybean (Glycine max) in Brazil by the nucleotide sequences of ITS-5.8S regions of rDNA. The 5.8S rDNA gene sequence (155 bp) was highly conserved among all isolates but differences in length and nucleotide sequence of the ITS1 and ITS2 regions were observed between soybean isolates and AG testers. The similarity of the nucleotide sequence among AG-1 IA isolates, causing foliar blight, was 95.1-100% and 98.5-100% in the ITS1 and ITS2 regions, respectively. The nucleotide sequence similarity among subgroups IA, IB and IC ranged from 84.3 to 89% in ITS1 and from 93.3 to 95.6% in ITS2. Nucleotide sequence similarity of 99.1% and 99.3-100% for ITS1 and ITS2, respectively, was observed between AG-4 soybean isolates causing hypocotyl rots and the AG-4 HGI tester. The similarity of the nucleotide sequence of the ITS-5.8S rDNA region confirmed that the R. solani Brazilian isolates causing foliar blight are AG-1 IA and isolates causing hypocotyl rot symptoms are AG-4 HGI. The ITS-5.8S rDNA sequence was not determinant for the identification of the AG-2-2 IIIB R. solani soybean isolate.

Caracterização espectral de estágios sucessionais de vegetação secundária arbórea em Altamira (PA), através de dados orbitais

Imagens TM/Landsat 5 (bandas TM1-TM5 e TM7) de Altamira, PA, de 07.02.1991 foram transformadas em reflectância de superfície usando o modelo 6S de correção atmosférica com o objetivo de caracterizar espectralmente formações florestais secundárias. Dados biofísicos (DAP, G e H) foram coletados de 16 parcelas em campo, os quais foram utilizados no cálculo da área basal total (GT) e da rugosidade do dossel (Rug) e no estabelecimento de correlações entre os valores de reflectância de superfície dessas mesmas parcelas. Os coeficientes de correlação mais elevados foram verificados nas relações com a banda TM7. Regressões lineares foram estabelecidas tendo como variável dependente os parâmetros biofísicos e como variável independente, os valores de reflectância de superfície da banda TM7, a partir das quais foram elaborados mapas temáticos representativos da distribuição espacial dos parâmetros biofísicos. A reflectância de superfície não foi sensível a valores pequenos de dados biofísicos. Apesar disso, a metodologia aqui empregada mostrou-se eficaz para estimativas remotas de dados dendrométricos.

Análise da dinâmica sazonal e separabilidade espectral de algumas fitofisionomias do cerrado com índices de vegetação dos sensores MODIS/TERRA e AQUA

Composições de 16 dias de índices de vegetação do sensor MODerate resolution Imaging Spectroradiometer (MODIS), com resolução espacial de 1km, a bordo dos satélites TERRA e AQUA, foram usadas para caracterizar a dinâmica sazonal em 2004 de cinco fitofisionomias de Cerrado e analisar a sua separabilidade espectral. Os índices Normalized Difference Vegetation Index (NDVI) e Enhanced Vegetation Index (EVI), calculados a partir dos dados dos sensores de ambas as plataformas e de uma base comum de pixels, foram comparados entre si. Os resultados indicaram que: (a) dentre as fitofisionomias estudadas, a Floresta Estacional decídua apresentou uma dinâmica sazonal muito marcante em função da perda de folhas da estação chuvosa para a seca (substancial redução nos índices) e do rápido verdejamento com o início da precipitação no final de outubro (rápido incremento de NDVI e EVI); (b) o NDVI mostrou maior variabilidade entre as classes de vegetação do que o EVI apenas na estação seca; (c) a discriminação entre as fitofisionomias melhorou da estação chuvosa para a seca; (d) o NDVI foi mais eficiente do que o EVI para discriminar as classes de vegetação na estação seca, ocorrendo o contrário na estação chuvosa; e (e) na maioria das datas selecionadas para estudo, não houve diferenças estatisticamente significativas entre os índices de vegetação gerados de ambas as plataformas, apesar das variações na qualidade dos pixels selecionados para as composições de 16 dias e na geometria de iluminação e de visada.

Relação entre índice de área foliar e frações de componentes puros do modelo linear de mistura espectral, usando imagens ETM+/Landsat

O índice de área foliar (IAF) é uma das mais importantes variáveis biofísicas da vegetação, estando relacionado diretamente com a evapotranspiração, com a produtividade da vegetação e com a interceptação da chuva pelo dossel. O objetivo deste trabalho foi analisar a relação do IAF de diversos tipos de cobertura do solo com Frações de Componentes Puros (FCPs) do Modelo Linear de Mistura Espectral (MLME). A área de estudo foi a microbacia hidrográfica do Ribeirão dos Marins, localizada no município de Piracicaba - SP. O IAF foi medido, no campo, com o equipamento LAI-2000, em 32 áreas com diferentes coberturas vegetais. A imagem utilizada foi do sensor ETM+ a bordo do satélite Landsat-7. No MLME, foram considerados três componentes puros (vegetação, solo e sombra), selecionados com o auxílio dos componentes principais. Como resultado, tem-se que o IAF variou de 0,47 a 4,48, quando consideradas todas as áreas. As relações do IAF com a fração do componente puro vegetação F VEG e com a fração do componente puro solo (F SOL) foram significativas, embora fracas. Ao considerar apenas dados de IAF de cana-de-açúcar, houve aumento da variação explicada tanto para F VEG como para F SOL, sugerindo que a estratificação da vegetação pelo tipo pode melhorar a estimativa do IAF.

Caracterização espectral de áreas de gramíneas forrageiras infectadas com a doença "mela-das-sementes da braquiária" por meio de imagens CCD/CBERS-2

Imagens CCD/CBERS-2, nas bandas espectrais CCD2, CCD3 e CCD4, dos anos de 2004 e 2005, de Mirante do Paranapanema - SP, foram transformadas em reflectância de superfície usando o modelo 5S de correção atmosférica e normalizadas radiometricamente. O objetivo principal foi caracterizar espectralmente áreas de pastagens de Brachiaria brizantha em fase de florescimento, isentas e infectadas com a doença "mela-das-sementes da braquiária", possibilitando a sua detecção por meio da comparação entre os valores de reflectância de superfície denominada de Fator de Reflectância Bidirecional de Superfície (FRBS). Teve-se, também, o objetivo de avaliar a eficácia das imagens CCD/CBERS-2 para a obtenção de respostas espectrais de pastagens. Os dosséis sadios e doentes da Brachiaria brizantha foram identificados por meio da análise dos valores de reflectância e dos dados observados no Índice de Estresse Hídrico Acumulativo Relativo da Cultura (ACWSI) obtidos na área de estudo. Os resultados indicaram que as principais diferenças foram a diminuição da reflectância na banda CCD3 e o aumento da reflectância na banda CCD4 nas áreas doentes. A metodologia empregada com o uso de dados do sensor CCD/CBERS-2, associados ao ACWSI, mostrou-se eficaz para discriminar dosséis infectados com a "mela-das-sementes da braquiária".

Classificação espectral de área plantada com a cultura da cana-de-açúcar por meio da árvore de decisão

O objetivo deste trabalho foi testar o classificador "árvore de decisão", em dados provenientes de sensores orbitais, para identificar área plantada com cana-de-açúcar, em diferentes épocas de plantio na Fazenda Boa Fé, localizada no Triângulo Mineiro, mais especificamente no município de Conquista, Minas Gerais. Acoplaram-se técnicas de Sensoriamento Remoto (SR) em um módulo de Sistema de Informação Geográfica (SIG), permitindo uma análise temporal do uso e ocupação do solo, especialmente com vistas a identificar e a monitorar as áreas agrícolas. Com base no cálculo do viés médio (VM), o presente estudo mostrou que, em áreas de cana-de-açúcar, onde a irrigação é frequente e ocorrem chuvas significativas que antecedem a passagem do satélite Landsat-5, os valores foram ligeiramente subestimados, com valor deste indicador de -0,13 ha. Foi verificado, também, que os valores de NDVI mais altos proporcionaram uma leve superestimativa dos resultados, com valores de viés médio variando de 0,04 a 0,23 ha. Conforme os resultados, o classificador árvore de decisão apresentou um grande potencial para o mapeamento das áreas cultivadas com cana-de-açúcar.

Occurrence of virulence-related sequences and phylogenetic analysis of commensal and pathogenic avian Escherichia coli strains (APEC)

The presence of iron uptake (irp-2, fyuA, sitA, fepC, iucA), adhesion (iha, lpfA O157/O141, lpfA O157/O154, efa, toxB) and invasion (inv, ial-related DNA sequences and assignment to the four main Escherichia coli phylogenetic groups (A, B1, B2 e D) were determined in 30 commensal E. coli strains isolated from healthy chickens and in 49 APEC strains isolated from chickens presenting clinical signs of septicemia (n=24) swollen head syndrome (n=14) and omphalitis (n=11) by PCR. None of the strains presented DNA sequences related to the inv, ial, efa, and toxB genes. DNA sequences related to lpfA O157/O154, iucA, fepC, and irp-2 genes were significantly found among pathogenic strains, where iucA gene was associated with septicemia and swollen head syndrome and fepC and irp-2 genes were associated with swollen head syndrome strains. Phylogenetic typing showed that commensal and omphalitis strains belonged mainly to phylogenetic Group A and swollen head syndrome to phylogenetic Group D. Septicemic strains were assigned in phylogenetic Groups A and D. These data could suggest that clonal lineage of septicemic APEC strains have a multiple ancestor origin; one from a pathogenic bacteria ancestor and other from a non-pathogenic ancestor that evolved by the acquisition of virulence related sequences through horizontal gene transfer. Swollen head syndrome may constitute a pathogenic clonal group. By the other side, omphalitis strains probably constitute a non-pathogenic clonal group, and could cause omphalitis as an opportunistic infection. The sharing of virulence related sequences by human pathogenic E. coli and APEC strains could indicate that APEC strains could be a source of virulence genes to human strains and could represent a zoonotic risk.

In silico phylogenetic and virulence gene profile analyses of avian pathogenic Escherichia coli genome sequences

Avian pathogenic Escherichia coli (APEC) infections are responsible for significant losses in the poultry industry worldwide. A zoonotic risk has been attributed to APEC strains because they present similarities to extraintestinal pathogenic E. coli (ExPEC) associated with illness in humans, mainly urinary tract infections and neonatal meningitis. Here, we present in silico analyses with pathogenic E. coli genome sequences, including recently available APEC genomes. The phylogenetic tree, based on multi-locus sequence typing (MLST) of seven housekeeping genes, revealed high diversity in the allelic composition. Nevertheless, despite this diversity, the phylogenetic tree was able to cluster the different pathotypes together. An in silico virulence gene profile was also determined for each of these strains, through the presence or absence of 83 well-known virulence genes/traits described in pathogenic E. coli strains. The MLST phylogeny and the virulence gene profiles demonstrated a certain genetic similarity between Brazilian APEC strains, APEC isolated in the United States, UPEC (uropathogenic E. coli) and diarrheagenic strains isolated from humans. This correlation corroborates and reinforces the zoonotic potential hypothesis proposed to APEC.

Phylogenetic analyses of Cladophora vagabunda (L.) C. Hoek (Cladophorales, Chlorophyta) from Brazil based on SSU rDNA sequences

The complete SSU rDNA was sequenced for 10 individuals of Cladophora vagabunda collected along the coast of Brazil. For C. rupestris (L.) Kütz. a partial SSU rDNA sequence (1634 bp) was obtained. Phylogenetic trees indicate that Cladophora is paraphyletic, but the section Glomeratae sensu lato including C. vagabunda from Brazil, Japan and France, C. albida (Nees) Kütz., C. sericea (Hudson) Kütz., and C. glomerata (L.) Kütz. is monophyletic. Within this group C. vagabunda is paraphyletic. The sequence identity for the SSU rDNA varied from 98.9% to 100% for the Brazilian C. vagabunda, and from 98.3% to 99.7% comparing the Brazilian individuals to the ones from France and Japan. Sequence identity of the Brazilian C. vagabunda to C. albida and C. sericea vary from 98.0% to 98.6%. The SSU rDNA phylogeny support partially the morphological characteristics presented by Brazilian populations of C. vagabunda. On the other hand, C. rupestris from Brazil does not group with C. rupestris from France, both sequences presenting only 96.9% of identity. The inclusion of sequences of individuals from Brazil reinforces the need of taxonomical revision for the genus Cladophora and for the complex C. vagabunda.

A 37-kb restriction map of the human immunoglobulin lambda variable locus, VB cluster, harboring four functional genes and two non-coding Vl sequences

The human immunoglobulin lambda variable locus (IGLV) is mapped at chromosome 22 band q11.1-q11.2. The 30 functional germline v-lambda genes sequenced untill now have been subgrouped into 10 families (Vl1 to Vl10). The number of Vl genes has been estimated at approximately 70. This locus is formed by three gene clusters (VA, VB and VC) that encompass the variable coding genes (V) responsible for the synthesis of lambda-type Ig light chains, and the Jl-Cl cluster with the joining segments and the constant genes. Recently the entire variable lambda gene locus was mapped by contig methodology and its one- megabase DNA totally sequenced. All the known functional V-lambda genes and pseudogenes were located. We screened a human genomic DNA cosmid library and isolated a clone with an insert of 37 kb (cosmid 8.3) encompassing four functional genes (IGLV7S1, IGLV1S1, IGLV1S2 and IGLV5a), a pseudogene (VlA) and a vestigial sequence (vg1) to study in detail the positions of the restriction sites surrounding the Vl genes. We generated a high resolution restriction map, locating 31 restriction sites in 37 kb of the VB cluster, a region rich in functional Vl genes. This mapping information opens the perspective for further RFLP studies and sequencing

Control of the rat angiotensin I converting enzyme gene by CRE-like sequences

We characterized the role of potential cAMP-responsive elements (CRE) in basal and in induced angiotensin converting enzyme (ACE) gene promoter activity in order to shed light on the regulation of somatic ACE expression. We identified stimulators and repressors of basal expression between 122 and 288 bp and between 415 and 1303 bp upstream from the transcription start site, respectively, using a rabbit endothelial cell (REC) line. These regions also contained elements associated with the response to 8BrcAMP. When screening for CRE motifs we found pCRE, a proximal sequence between 209 and 222 bp. dCRE, a distal tandem of two CRE-like sequences conserved between rats, mice and humans, was detected between 834 and 846 bp. Gel retardation analysis of nuclear extracts of REC indicated that pCRE and dCRE bind to the same protein complexes as bound by a canonical CRE. Mutation of pCRE and dCRE in REC established the former as a positive element and the latter as a negative element. In 293 cells, a renal cell line, pCRE and dCRE are negative regulators. Co-transfection of ATF-2 or ATF-2 plus c-Jun repressed ACE promoter activity, suggesting that the ACE gene is controlled by cellular stress. Although mapping of cAMP responsiveness was consistent with roles for pCRE and dCRE, mutation analysis indicated that they were not required for cAMP responsiveness. We conclude that the basal activity of the somatic ACE promoter is controlled by proximal and distal CREs that can act as enhancers or repressors depending on the cell context.

Automatic recognition of fish from video sequences

The problem of automatic recognition of the fish from the video sequences is discussed in this Master’s Thesis. This is a very urgent issue for many organizations engaged in fish farming in Finland and Russia because the process of automation control and counting of individual species is turning point in the industry. The difficulties and the specific features of the problem have been identified in order to find a solution and propose some recommendations for the components of the automated fish recognition system. Methods such as background subtraction, Kalman filtering and Viola-Jones method were implemented during this work for detection, tracking and estimation of fish parameters. Both the results of the experiments and the choice of the appropriate methods strongly depend on the quality and the type of a video which is used as an input data. Practical experiments have demonstrated that not all methods can produce good results for real data, whereas on synthetic data they operate satisfactorily.