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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
21

Amarelo da ameixeira: caracterização molecular do fitoplasma e modelo de colonização do hospedeiro / Plum yellow: molecular characterization of the phytoplasma and colonization model of the host

Daniela Flores 02 October 2013 (has links)
No Brasil, o cultivo de ameixeira tem atingido significativa importância econômica em termos de rentabilidade. Embora rentável, a cultura exige cuidados constantes em relação aos danos provocados por doenças. Entre elas, o \'amarelo\', causado por fitoplasmas, tem se mostrado como uma doença de relevância nos países produtores desta fruta. Em pomares comerciais localizados em Paranapanema-SP, foram observadas ameixeiras (Prunus salicina) exibindo sintomas típicos de \'amarelos\', caracterizados por superbrotamento de ramos, redução no comprimento de entrenós, além de amarelecimento, deformação e redução do limbo foliar. Com objetivo de identificar o fitoplasma e determinar o modelo de colonização do hospedeiro pelo patógeno foi desenvolvido o presente estudo. Para isto, em três pomares, foram amostradas plantas sintomáticas, assintomáticas e sadias, pertencentes às variedades Gulfblaze e Azteca. O DNA total foi extraído de folhas e ramos e usado em duplo PCR com os iniciadores R16mF2/mR1 e R16F2n/R2, visando a detecção do fitoplasma. Os resultados confirmaram a presença de fitoplasma pela amplificação de fragmentos de DNA de 1,2 kb do gene 16S rRNA. Os produtos de PCR gerados por dez isolados de fitoplasmas de cada variedade foram clonados e três clones de cada isolado foram sequenciados. Uma vez que nenhum polimorfismo foi encontrado, uma sequência consenso foi selecionada para cada isolado e um isolado foi escolhido como representativo para cada variedade. Estas sequências foram designadas por PlY-BR01 e PlY-BR02, com 1.246 pb, e depositadas no GenBank sob os números de acesso KF499086 e KF499087, respectivamente. A sequência do 16S rRNA do fitoplasma da ameixeira apresentou 100% de identidade com o fitoplasma representante do grupo 16SrI-B (AY265208). A análise de RFLP virtual e o cálculo do coeficiente de similaridade, permitiram classificar o fitoplasma da ameixeira no grupo 16SrI-B. A análise filogenética revelou que o mesmo está estritamente relacionado ao subgrupo 16SrI-B. No estudo da colonização das plantas pelo patógeno, amostras da copa e raiz foram coletadas mensalmente, durante um ano, de plantas infectadas das variedades Gulfblaze e Azteca. O DNA total foi extraído e submetido ao PCR em Tempo Real com os iniciadores UniRNAForward/UniRNAReverse, para quantificar o fitoplasma nos tecidos do hospedeiro. O fitoplasma foi detectado tanto em amostras de copa como de raiz, em todas as árvores infectadas de ambas as variedades. A concentração do mesmo nos tecidos vegetais variou de 5,77x105 a 1,93x109 e 6,18x105 a 3,92x109 N°cópias/100 ng de DNA total, na copa e na raiz, respectivamente. O experimento mostrou uma flutuação sazonal na concentração do fitoplasma, sendo as maiores concentrações encontradas no verão, embora o fitoplasma tenha permanecido na parte aérea do hospedeiro mesmo no período mais frio, que compreende a fase de dormência da planta. Com base nestes resultados, ficou evidente que as amostras retiradas da copa e coletadas durante o período mais quente do ano são as mais adequadas para os procedimentos de detecção do fitoplasma, visando confirmar a diagnose feita com base na sintomatologia. / In Brazil, the cultivation of plum trees has achieved significant economic importance in terms of profitability. Although profitable, the culture demands attention in relation to damages provoked by diseases. Among them, the \'yellow\', caused by phytoplasmas, is a relevant disease present in the plum producing countries. In commercial orchards located in Paranapanema-SP region, were observed plum trees (Prunus salicina) exhibiting typical symptoms of \'yellow\', characterized by intense shoot proliferation, shortened internodes, generalized yellowing, leaf malformation and small leaves. The present study was conducted in order to identify the phytoplasma associated with symptomatic plants and determine the pattern of host colonization by the pathogen. Therefore, were sampled symptomatic, asymptomatic and healthy plants of the varieties Gulfblaze and Azteca, grown in three commercial orchards. Total DNA was extracted from leaves and shoots and submitted to nested PCR primed with R16mF2/mR1 and R16F2n/R2, in order to detect phytoplasma. The results confirmed the presence of phytoplasma by amplification of DNA fragments of 1.2 kb from 16S rRNA gene. The PCR products generated by ten phytoplasma isolates of each variety were cloned and three clones of each isolate were sequenced. Since no polymorphism was found, a consensus sequence was selected for each isolate and an isolate was chosen as representative for each variety. These sequences were designated by PlYBR01 and PlY-BR02, with 1,246 bp, and deposited in GenBank under the accession numbers KF499086 and KF499087, respectively. The sequence of the 16S rRNA of the phytoplasma founded in plum trees showed 100% identity with the phytoplasma representative of 16SrI-B group (AY265208). The virtual RFLP analysis and the calculation of the similarity coefficient, allowed classify the phytoplasma present in plum trees as a member of 16SrI-B group. Phylogenetic analysis supported that this phytoplasma is closed related to the 16SrI-B subgroup. In the study about plant colonization, the samples from leaves, shoots and root were monthly collected, for one year, from the infected plants of the varieties Gulfblaze and Azteca. Total DNA was extracted and submitted to the Real Time PCR with primers UniRNAForward/UniRNAReverse, in order to quantify phytoplasma in host tissues. The phytoplasma was detected in both aerial parts and roots in all infected trees of the two varieties. The concentration of the pathogen in plant tissues ranged from 5.77 x105 to 1.93 x109 and 6.18 x105 to 3.92 x109 N°copies/100ng total DNA in aerial parts and roots, respectively. The experiment showed a seasonal fluctuation in the concentration of the phytoplasma in plants, with the highest concentrations found in summer, although the phytoplasma have remained in the shoots of the host even in the coldest period, comprising the dormant phase of the plant. Based on these results, it was evident that the samples collected from the aerial parts and during the hottest period of the year are the most appropriate for detection of phytoplasma to confirm the diagnosis based on symptomatology.
22

Revisão taxonômica das abróteas do gênero Urophycis Gill, 1863 no Atlântico Sul (Gadiformes: Gadidae) / Taxonomic review of codlings of genus Urophycis Gill, 1863 in the South Atlantic (Gadiformes: Gadidae)

Lemes, Paola Cristina Roque 05 May 2017 (has links)
Urophycis Gill, 1863 é um gênero de peixes marinhos demersais de médio porte, popularmente conhecidos como abróteas, distribuídos ao longo da costa do Atlântico Ocidental, do Canadá à Argentina. Atualmente são reconhecidas sete espécies válidas dentro deste gênero. Apesar de sua importância comercial, até então poucos estudos taxonômicos e biogeográficos haviam sido realizados com estas espécies. Três espécies foram descritas do Atlântico Sul ocidental: U. brasiliensis (Kaup, 1858), descrita de Motevideo, Uruguai, U. latus Miranda Ribeiro, 1903 e Urophycis mystacea Miranda Ribeiro, 1903, ambas descritas do Rio de Janeiro, Brasil. Urophycis mystacea é morfologicamente semelhante à U. cirrata (Goode & Bean, 1896), descrita do Golfo do México nos Estados Unidos, sendo frequentemente confundidas. Os objetivos desta contribuição foram (1) redescrever U. brasiliensis, e alocar U. latus como um sinônimo júnior ; (2) redescrever U. cirrata e alocar U. mystacea como um sinônimo júnior; (3) estabelecer diagnoses e mapas de distribuição atualizados para ambas espécies. Tecidos e exemplares foram obtidos com pescadores e em coleções científicas. Foram utilizados métodos tradicionais de morfologia para comparação dos exemplares, além de técnicas de identificação molecular utilizando o gene mitocondrial citocromo oxidase I (COI). Testes para a amplificação da região controle do DNA mitocondrial de U. brasiliensis foram realizados e discutidos. / Urophycis Gill, 1863 is a genus of marine, demersal, mid-sized fish, popularly known as codlings or hakes, and distributed along the the Western Atlantic coast, from Canada to Argentina. Seven species are currently recognized as valid within this genus. Despite its economical importance, a few taxomomic an biogeographic studies had been carried out with those species. Three species were described from western South Atlantic: U. brasiliensis (Kaup, 1858), described from Montevideo, Uruguay, U. latus Miranda Ribeiro, 1903 and Urophycis mystacea Miranda Ribeiro, 1903, both described from Rio de Janeiro, Brazil. Urophycis mystacea is morfologically similar to U. cirrata (Goode & Bean, 1896), described from Gulf of Mexico, USA, and these species are repeteadly confused. The objectives of this contributions were (1) redescribe U. brasilienis, placing U. latus as a junior synonym; (2) redescribe U. cirrata, placing U. mystacea as a junior synonym; (3) provide updated diagnosis and maps of distributions for both species. Tissues and specimens were obtained from fisherman and scientific collections. Traditional morphological methods were used to compare the specimens, besides molecular identification techniques using the mithocondrial gene citochrome c oxidadse I (COI). Testst for amplification of the DNA mitochondrial control region in U. brasiliensis were performed and discussed.
23

Revisão taxonômica das abróteas do gênero Urophycis Gill, 1863 no Atlântico Sul (Gadiformes: Gadidae) / Taxonomic review of codlings of genus Urophycis Gill, 1863 in the South Atlantic (Gadiformes: Gadidae)

Paola Cristina Roque Lemes 05 May 2017 (has links)
Urophycis Gill, 1863 é um gênero de peixes marinhos demersais de médio porte, popularmente conhecidos como abróteas, distribuídos ao longo da costa do Atlântico Ocidental, do Canadá à Argentina. Atualmente são reconhecidas sete espécies válidas dentro deste gênero. Apesar de sua importância comercial, até então poucos estudos taxonômicos e biogeográficos haviam sido realizados com estas espécies. Três espécies foram descritas do Atlântico Sul ocidental: U. brasiliensis (Kaup, 1858), descrita de Motevideo, Uruguai, U. latus Miranda Ribeiro, 1903 e Urophycis mystacea Miranda Ribeiro, 1903, ambas descritas do Rio de Janeiro, Brasil. Urophycis mystacea é morfologicamente semelhante à U. cirrata (Goode & Bean, 1896), descrita do Golfo do México nos Estados Unidos, sendo frequentemente confundidas. Os objetivos desta contribuição foram (1) redescrever U. brasiliensis, e alocar U. latus como um sinônimo júnior ; (2) redescrever U. cirrata e alocar U. mystacea como um sinônimo júnior; (3) estabelecer diagnoses e mapas de distribuição atualizados para ambas espécies. Tecidos e exemplares foram obtidos com pescadores e em coleções científicas. Foram utilizados métodos tradicionais de morfologia para comparação dos exemplares, além de técnicas de identificação molecular utilizando o gene mitocondrial citocromo oxidase I (COI). Testes para a amplificação da região controle do DNA mitocondrial de U. brasiliensis foram realizados e discutidos. / Urophycis Gill, 1863 is a genus of marine, demersal, mid-sized fish, popularly known as codlings or hakes, and distributed along the the Western Atlantic coast, from Canada to Argentina. Seven species are currently recognized as valid within this genus. Despite its economical importance, a few taxomomic an biogeographic studies had been carried out with those species. Three species were described from western South Atlantic: U. brasiliensis (Kaup, 1858), described from Montevideo, Uruguay, U. latus Miranda Ribeiro, 1903 and Urophycis mystacea Miranda Ribeiro, 1903, both described from Rio de Janeiro, Brazil. Urophycis mystacea is morfologically similar to U. cirrata (Goode & Bean, 1896), described from Gulf of Mexico, USA, and these species are repeteadly confused. The objectives of this contributions were (1) redescribe U. brasilienis, placing U. latus as a junior synonym; (2) redescribe U. cirrata, placing U. mystacea as a junior synonym; (3) provide updated diagnosis and maps of distributions for both species. Tissues and specimens were obtained from fisherman and scientific collections. Traditional morphological methods were used to compare the specimens, besides molecular identification techniques using the mithocondrial gene citochrome c oxidadse I (COI). Testst for amplification of the DNA mitochondrial control region in U. brasiliensis were performed and discussed.
24

Ecological investigation of a new host-parasite relationship : <i>Parelaphostrongylus odocoilei</i> in thinhorn sheep (<i>Ovis dalli</i>)

Jenkins, Emily Joan 20 September 2005
Discovery of a new host-parasite relationship, <i>Parelaphostrongylus odocoilei</i> in Dalls sheep (<i>Ovis dalli dalli</i>) in the Canadian North, prompted the first investigation of the geographic distribution, pathogenesis, ecology and epidemiology of this parasite, as well as the related protostrongylid <i>Protostrongylus stilesi</i>, at Subarctic latitudes (60-65ºN). All protostrongylid parasites have an indirect life-cycle, where first-stage larvae are shed in the feces of a mammalian definitive host, penetrate the foot of a gastropod intermediate host, and develop to infective third-stage larvae. <p> Protostrongylid larvae were recovered from over 2000 fecal samples from thinhorn sheep (<i>Ovis dalli</i>) and other hosts for <i>P. odocoilei</i> and <i>P. stilesi</i> across northwestern North America (38-69 ºN). Through novel application of molecular techniques to identify morphologically indistinguishable first-stage larvae, new records for <i>P. odocoilei</i> were established at 20 locations. This provided insight into the historical origins and biogeography of this new host-parasite relationship, and greatly expanded the known geographic range of both protostrongylids. <p> Clinical effects, including a neurological syndrome, were described in five thinhorn sheep experimentally infected with <i>P. odocoilei</i>. Neural and respiratory pathology in these five sheep were compared with over 50 wild Dalls sheep from a population naturally infected with <i>P. odocoilei</i> and <i>P. stilesi</i>. In the end stages, diffuse verminous interstitial pneumonia associated with P. odocoilei led to respiratory failure, and may have acted as a predisposing factor for bacterial pneumonia, which caused sporadic mortalities in this wild population. <p> At Subarctic latitudes, seasonal patterns in host and parasite availability, including larval shedding by Dalls sheep and larval development in experimentally infected gastropods, suggested that lambs become infected with <i>P. odocoilei</i> in a narrow seasonal window in their first fall on winter range. In combination with laboratory experiments, a degree day model for temperature-dependent larval development was developed, validated, and applied to describe and predict the effects of climate warming on protostrongylid parasites of thinhorn sheep in northern North America. In a future of climate warming, the narrow seasonal window for parasite development and transmission would be significantly extended, leading to amplification of populations of <i>P. odocoilei</i> and <i>P. stilesi</i> in endemic regions, and possibly range expansion of <i>P. odocoilei</i>. This may have consequences for the health of thinhorn sheep, as well as other wildlife that are important resources in the Canadian North.
25

Ecological investigation of a new host-parasite relationship : <i>Parelaphostrongylus odocoilei</i> in thinhorn sheep (<i>Ovis dalli</i>)

Jenkins, Emily Joan 20 September 2005 (has links)
Discovery of a new host-parasite relationship, <i>Parelaphostrongylus odocoilei</i> in Dalls sheep (<i>Ovis dalli dalli</i>) in the Canadian North, prompted the first investigation of the geographic distribution, pathogenesis, ecology and epidemiology of this parasite, as well as the related protostrongylid <i>Protostrongylus stilesi</i>, at Subarctic latitudes (60-65ºN). All protostrongylid parasites have an indirect life-cycle, where first-stage larvae are shed in the feces of a mammalian definitive host, penetrate the foot of a gastropod intermediate host, and develop to infective third-stage larvae. <p> Protostrongylid larvae were recovered from over 2000 fecal samples from thinhorn sheep (<i>Ovis dalli</i>) and other hosts for <i>P. odocoilei</i> and <i>P. stilesi</i> across northwestern North America (38-69 ºN). Through novel application of molecular techniques to identify morphologically indistinguishable first-stage larvae, new records for <i>P. odocoilei</i> were established at 20 locations. This provided insight into the historical origins and biogeography of this new host-parasite relationship, and greatly expanded the known geographic range of both protostrongylids. <p> Clinical effects, including a neurological syndrome, were described in five thinhorn sheep experimentally infected with <i>P. odocoilei</i>. Neural and respiratory pathology in these five sheep were compared with over 50 wild Dalls sheep from a population naturally infected with <i>P. odocoilei</i> and <i>P. stilesi</i>. In the end stages, diffuse verminous interstitial pneumonia associated with P. odocoilei led to respiratory failure, and may have acted as a predisposing factor for bacterial pneumonia, which caused sporadic mortalities in this wild population. <p> At Subarctic latitudes, seasonal patterns in host and parasite availability, including larval shedding by Dalls sheep and larval development in experimentally infected gastropods, suggested that lambs become infected with <i>P. odocoilei</i> in a narrow seasonal window in their first fall on winter range. In combination with laboratory experiments, a degree day model for temperature-dependent larval development was developed, validated, and applied to describe and predict the effects of climate warming on protostrongylid parasites of thinhorn sheep in northern North America. In a future of climate warming, the narrow seasonal window for parasite development and transmission would be significantly extended, leading to amplification of populations of <i>P. odocoilei</i> and <i>P. stilesi</i> in endemic regions, and possibly range expansion of <i>P. odocoilei</i>. This may have consequences for the health of thinhorn sheep, as well as other wildlife that are important resources in the Canadian North.
26

Identificação molecular e caracterização de peixes do gênero Zungaro (Siluriformes: Pimelodidae) de diferentes bacias hidrográficas

Pires, Antonio Augusto Adami 30 May 2014 (has links)
Made available in DSpace on 2016-06-02T20:21:37Z (GMT). No. of bitstreams: 1 6546.pdf: 2423797 bytes, checksum: 0ce860bab03667c9b7286a2c7da3f9a1 (MD5) Previous issue date: 2014-05-30 / Financiadora de Estudos e Projetos / The Genus Zungaro, Bleeker, 1858 (Siluriformes: Pimelodidae), comprise a group called Jaú. This group is divided in two species: Zungaro jahu located in Paraná- Paraguai basins and Zungaro zungaro located in Amazonas-Orinoco basins. Both species are known because they are the biggest catfish in South America. In Juruena River, smaller catfishes are found by local fishermen and because of that it´s very hard to classify these species. During this work Juruena population was genetically analyzed and these molecular datum were compared with other population to know if they are the same specie or not. Samples of fish came from Juruena, Amazonas, Solimões, Madeira, Xingu, Meta, Miranda, Taquari e Cuiabá were collected and kept in the heap of tissue bank in the Molecular Biodiversity and Conservation Laboratory (LabBMC). After isolating the DNA according to Sambrook protocol, mitochondrial sequences of cytochrome oxidase I (COI), cytochrome b (Cytb) and D-Loop were amplified by PCR. After the sequencing and edition, the results were compared to the databases BOLD and GenBank. Phylogenetic trees based on different algorithms were built. The species of Amazonas-Orinoco basins (Z. zungaro) and Paraná-Paraguai basins (Z. jahu) showed a considerable genetic distance highlighting a population structuring. Individual from Juruena River were different from the others following the model K2p 2.4% (COI) and 5% (D-Loop). The results showing an evidence of three different species: Z. zungaro, Z. jahu and possible new species in the Juruena River. Besides, specimens from Xingu River demonstrated a high genetic distance from the others, being necessary further analyses. / O gênero Zungaro, Bleeker, 1858 (Siluriformes: Pimelodidae), pertence a um grupo de peixes popularmente denominados jaús, composto por duas espécies válidas: Zungaro jahu que ocorre na bacia dos rios Paraná-Paraguai e Zungaro zungaro encontrada na bacia dos rios Amazonas e Orinoco. Este gênero é conhecido por compreender um dos maiores bagres da América do Sul. Em especial na região do Mato-Grosso, no rio Juruena. Estas espécies são conhecidas por serem os maiores bagres da América do Sul. No rio Juruena (MT), indivíduos de jaús adultos tem sido capturados por pescadores com um tamanho menor do que o descrito na literatura e o permitido para pesca, levantando incertezas sobre a classificação desta espécie nesta região. Neste trabalho foram analisadas populações do Juruena e os dados moleculares obtidos foram comparados com os de outras populações de jaús para saber se tratam da mesma espécie. Amostras de peixes dos rios Juruena, Amazonas, Solimões, Madeira, Xingu, Meta, Miranda, Taquari e Cuiabá foram obtidas e armazenadas no acervo do Banco de Tecidos do laboratório de Biodiversidade Molecular e Conservação (LabBMC). Após isolar o DNA segundo o protocolo de Sambrook, fragmentos mitocondriais dos genes Citocromo Oxidase I (COI) e Citocromo b (Cytb) e da região controle D-Loop foram amplificados via PCR. Após o sequenciamento e edição das sequências, os resultados obtidos foram comparados com sequências disponíveis nas bases de dados BOLD e GenBank. Árvores filogenéticas baseadas em diferentes algoritmos foram construídas. As espécies das bacias Amazonas-Orinoco (Z. zungaro) e Paraná-Paraguai (Z. jahu) apresentaram uma distância genética considerável, evidenciando estruturação populacional. Indivíduos do rio Juruena apresentaram-se diferenciados das demais espécies válidas para o gênero com valor de distância genética segundo o modelo K2p de 2.4% (COI) e 5%(D-Loop). Estes resultados apontam a evidência da existência de no mínimo três espécies para o Gênero: Z. zungaro, Z. jahu e uma possível nova espécie do rio Juruena. Espécimes do rio Xingu também demostraram alta distância genética com relação às demais populações analisadas, necessitando uma investigação mais profunda.
27

Studium mikrokrystalických inkluzí v jednobuněčných řasách pomocí Ramanovy mikroskopie / Microcrystalline inclusions in microalgae studied via Raman microscopy

Suja, Matyáš January 2019 (has links)
Many freshwater, terrestrial or marine microalgae contain various microcrystalline inclusions that they use in their life cycle. However, the identification of the molecular composition of these inclusions via many physical or chemical methods is often very difficult and susceptible to many measurement errors. Therefore, the chemical composition of these microcrystals in many microalgae has not been determined at all or may be incorrect. One of the high precision methods capable of determining the composition of microcrystalline bodies within microalgae is Raman confocal microscopy. This very promising method of optical vibrational spectroscopy allows rapid and non-destructive molecular analysis of objects. Raman microscopy does not require chemical extraction, modification or other color marking or staining of the sample. Therefore, it can directly measure living cells at various stages of their natural development. The chemical composition of the sample is then characterized by its corresponding Raman vibrational spectrum. The aim of this diploma thesis is to determine the presence of microcrystals in different species of microalgae, study the conditions of their occurrence and identify their chemical composition via Raman microscopy.
28

Sele??o e identifica??o de Trichoderma spp. e potencial para produ??o de enzimas industriais / Selection and identification of Trichoderma spp. And Potential for Industrial Enzymes production

Francisco, Michele Rodrigues 20 May 2016 (has links)
Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-05-12T11:13:08Z No. of bitstreams: 1 2016 - Michele Rodrigues Francisco.pdf: 1285744 bytes, checksum: 598c2fe8a98038b9745debaf13625959 (MD5) / Made available in DSpace on 2017-05-12T11:13:08Z (GMT). No. of bitstreams: 1 2016 - Michele Rodrigues Francisco.pdf: 1285744 bytes, checksum: 598c2fe8a98038b9745debaf13625959 (MD5) Previous issue date: 2016-05-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Trichoderma spp. They are not pathogenic fungi and develop in substrates and diverse environmental conditions. Identification based only on morphological characteristics is considered complex, due to the genetic diversity of the species. Therefore, the taxonomy of the genus Trichoderma is performed with the aid of other analyzes such as biochemical and molecular changes. Some species are capable of producing high amounts of extracellular enzymes involved in the hydrolysis of polysaccharides are thus considered fungi with high potential in the production of hydrolytic enzymes. The aim of this study was to select and identify the species of the genus Trichoderma spp. And evaluate the production of industrial enzymes these isolates. They identified 58 isolates of Trichoderma spp fungi. soil and plants by morphological and molecular analysis and was also evaluated the enzymatic production capacity. Enzyme technology is a promising area in new technologies for synthesis of compounds with high added value, where biocatalisados industrial processes have a lower environmental impact, lower energy cost, because the enzymes are biodegradable substances and have specificity to minimize the undesirable effects. In this work, we used PDA culture medium for development of fungi. Morphological identification is achieved using the identification key according to Samuels et al. (2015) Molecular and held by DNA extraction from the mycelium of fungi in accordance with CTAB protocol. For enzyme analysis, qualitative assays were performed, used specific means of solid culture to evaluate the production of hydrolytic enzymes: Where the culture media containing as a substrate for development and enzyme production of Trichoderma spp. skimmed milk production peptidase; soluble starch for the production of amylase, cellulase production carboxymethylcellulose, citrus pectin for the production of pectinase and yeast extract, dextrose, ferrous sulfate, ammonium, magnesium, manganese and potassium chloride for the production of phytase. According to the results, the morphological analysis identified 4 species such as T. atroviride, T .asperelloide, T. harzianum, T. longibrachiatum, these species were verified by molecular analysis. In relation to the enzyme potential, the result was negative for the activity of peptidase enzymes, cellulase and pectinase and positive for amylase, and the activity of the enzyme phytase, only an isolated was positive, this was identified according to morphological analysis as It is the species T. longibrachiatum / Os Trichoderma spp. n?o s?o fungos patog?nicos e se desenvolvem em diversificados substratos e condi??es ambientais. Sua identifica??o baseada apenas em caracter?sticas morfol?gicas ? considerada complexa, por conta da diversidade gen?tica das esp?cies. Portanto, a taxonomia do g?nero Trichoderma ? realizada com o aux?lio de outras an?lises, tais como bioqu?micas e moleculares. Algumas esp?cies s?o capazes de produzir elevadas quantidades de enzimas extracelulares envolvidas na hidr?lise de polissacar?deos, sendo assim considerados fungos com alto potencial na produ??o de enzimas hidrol?ticas. O objetivo deste estudo foi selecionar e identificar as esp?cies do g?nero Trichoderma spp. e avaliar a produ??o de enzimas industriais destes isolados. Foram identificados 58 isolados de fungos Trichoderma spp. de solo e plantas por an?lise morfol?gica, molecular e tamb?m foi avaliado a capacidade de produ??o enzim?tica. A Tecnologia enzim?tica ? uma ?rea promissora dentro das novas tecnologias para s?ntese de compostos que possuem alto valor agregado, onde os processos industriais biocatalisados apresentam menor impacto ambiental, menor custo de energia, pois as enzimas s?o subst?ncias biodegrad?veis e possuem especificidade que minimizam os efeitos indesej?veis. Neste trabalho, foi utilizado meio de cultura BDA para desenvolvimento dos fungos. A identifica??o morfol?gica foi realizada com aux?lio da chave de identifica??o de acordo com Samuels et al. (2015) e molecular, realizada atrav?s da extra??o de DNA do mic?lio dos fungos, de acordo com protocolo CTAB. Para as an?lises enzim?ticas, foram realizados ensaios qualitativos, utilizado meio de cultura s?lido espec?fico para avaliar a produ??o de enzimas hidrol?ticas: Onde os meios de cultura continham como substrato para desenvolvimento e produ??o enzim?tica dos isolados de Trichoderma spp. leite desnatado para produ??o de peptidase; amido sol?vel para produ??o de amilase, carboximetilcelulose para produ??o de celulase, pectina c?trica para produ??o de pectinases e extrato de levedura, dextrose, sulfato ferroso, am?nio, magn?sio, mangan?s e cloreto pot?ssio para produ??o de fitase. De acordo com os resultados, a an?lise morfol?gica identificou 4 esp?cies, tais como: T. atroviride, T. asperelloide, T. harzianum, T. longibrachiatum, estas esp?cies foram confirmadas por an?lise molecular. J? com rela??o ao potencial enzim?tico, o resultado foi negativo para a atividade das enzimas peptidase, celulase e pectinase e positivo para amilase, e para a atividade da enzima fitase, somente um isolado apresentou resultado positivo, este foi identificado de acordo com an?lise morfol?gica como sendo da esp?cie T. longibrachiatum
29

Fauna karabida (Coleoptera; Carabidae) u razliĉitimagroekološkim uslovima Vojvodine

Popović Aleksandra 12 May 2015 (has links)
<p>Carabide pripadaju kosmopolitskoj grupi insekata, sa preko 40.000 vrsta &scaron;irom Sveta,<br />od tog broja 2700 vrsta je registrovano u Evropi. Trenutno se smatra da su trĉuljci<br />najrazvijenija familija podreda Adephaga. Desile su se mnoge izmene u sistematici<br />familije Carabidae od prvog kljuĉa za determinaciju &ldquo;Royal Entomological Society<br />handbook&ldquo; (Lindroth,1974). Prvi znaĉajan doprinos razumevanja ekologije, taksonomije<br />i rasprostranjenosti karabida naĉinio je Carl Heinz Lindroth, koji je pokrio veći deo<br />severne Evrope kao i razvijene delove Severne Amerike, rad Lindrota pratio je Thiele<br />(1977). Kasniji rad na identifikaciji karabida u Evropi izmenio je kljuĉ za determinaciju<br />vrsta centralnog dela Evrope. Takodje, napravljen je veći broj ilustrovanih kljuĉeva za<br />determinaciju karabida na prostoru Francuske. Karabide su tipiĉni polifagni predatori,<br />veoma znaĉajni za odrţanje agroekosistema i drugih ekosistema zbog ĉega su ĉest<br />predmet istraţivanja. Pored toga &scaron;to su poznati kao korisni insekti, odnosno regulatori<br />brojnosti &scaron;tetnih populacija, neki predstavnici familije Carabidae mogu biti i &scaron;tetoĉine<br />biljaka kako gajenih tako i korovskih, ĉime se uloga pripadnika ove familije znaĉajno<br />uvećava. Cilj ovog rada jeste determinacija imaga familije Carabidae i njihova potvrda<br />putem molekularnih analiza (PCR), kao i utvrĊivanje kvalitativnog i kvantitativnog<br />sastava trĉuljaka u poljima p&scaron;enice, kukuruza i &scaron;ećerne repe na podruĉju, Beĉeja,<br />Maglića i Rimskih &Scaron;anĉeva, tokom 2010. i 2011. godine. TakoĊe, je posmatrana<br />dominantnost, abundantnost, stepen faunistiĉke sliĉnosti karabida i uticaja<br />agrometeorolo&scaron;kih uslova sredine na njihovu brojnost i sastav vrsta. Tokom<br />dvogodi&scaron;njih istraţivanja u tri napred navedena useva i na sva tri lokaliteta, metodom &bdquo;<br />Barber posuda&ldquo; sakupljeno je ukupno 4.420 jedinki familije Carabidae. Kada<br />posmatramo brojnost po godinama, u prvoj godini, prikupljeno je vi&scaron;e jedinki tj. 2803,<br />koje su svrstane u 51 vrstu, dok je u 2011.g., brojnost bila skoro duplo manja, a<br />zabeleţeno je svega 1.617 jedinki, odnosno identifikovano je 47 vrsta, &scaron;to je u vezi sa uslovima sredine u datim godinama istraţivanja. TakoĊe, na osnovu dobijenih rezultata moţe se zakljuĉiti da je struktura karabida zavisna od vladajućih biotiĉkih i abiotiĉkih faktora, te kombinacijom ovih faktora stvaraju se specifiĉni mikroklimatski uslovi, a sa tim i karakteristiĉna fauna. Nakon uraĊenih molekularnih analiza, kod vrsta Calathus fuscipes, Poecilus versicolor, Amara aenea, Calathus ambiguus, Anchomenus dorsalis, Dolichus halensis, Pterostichus cupreus, Harpalus distinguendus, Harpalus rufipes, Laemostenus terricola, Pterostichus melanarius, Harpalus dimidiatus, Anisodactylus binotatus, Trechus quadristriatus, Carabus coriaceus, Carabus cancellatus, BLAST analiza je potvrdila status vrste koja je primarno identifikovana na osnovu morfolo&scaron;kih karakteristika. Vrste, kod kojih je utvrĊena visoka stopa sliĉnosti sa drugim vrstama iz istog roda i za koje nema referentnih sekvenci u banci gena, kao &scaron;to su Pterostichus (Poecilus) sericeus, Harpalus azureus, Pterostichus incommodus, Harpalus griseus, Pterostichus (Cophosus) cylindricus, Pterostichus vernalis, Pterostichus (Feronidius) melas i Calosoma auropunctatum predstavljaju doprinos svetskoj barkoding bazi podataka.</p> / <p>Carabidae belong to cosmopolitan group of insects, with over 40,000 species worldwide, where out of that number 2700 species is registered in Europe. It is currently considered that ground beetles are the most developed family of the sub-order Adephagae. There have been many changes in the systematic approach of the Carabidae family since the first determination key called &quot;Royal Entomological Society Handbook&quot; (Lindroth, 1974). The first significant contribution to the understanding of the ecology, taxonomy and distribution of Carabidae was made by Carl Heinz Lindroth that covered the greater part of the Northern Europe as well as the developed parts of Northern America; Lindroth&rsquo;s work was followed by Thiele (1977). Subsequent work on the identification of Carabidae in Europe changed the key for determination of the species of central Europe. Moreover, there have been a larger number of illustrated keys for determination of Carabidae on the territory of France. Carabidae are the typical polyphagous predators, very important for the maintenance of agro ecosystems and other ecosystems therefore they are often the subject of research. In addition to being known as beneficial insects, that is regulators of the number of pest populations, some representatives of the Carabidae family can also be pests of cultivated plants as well as weeds; therefore the role of the members of this family is increased significantly. The aim of this study is to determine the adults of the Carabidae family and their confirmation in molecular analysis (PCR), as well as to determine the qualitative and quantitative composition of ground beetles in fields of wheat, maize and sugar beet on the territory of Beĉej, Maglić and Rimski &Scaron;anĉevi, during 2010 and 2011. Furthermore, the dominance, abundance, degree of faunal similarity of Carabidae and the impact of agro-meteorological conditions of the environment on the number and composition of species were observed. During the two-year research in the three above-mentioned crops and in all three locations, using the method of &quot;Barber traps&quot; a total of 4,420 individuals of the Carabidae family were collected. When we look at the strength throughout years, in the first year more individuals were collected, that is 2,803 in number, which are classified into 51 species, while in 2011 the number was almost half that, and only 1,617 individuals were recorded, and 47 species identified, which is</p><p>related to environmental conditions in given years of study. Moreover, based on the<br />obtained results it can be concluded that the composition of Carabidae is dependent on<br />the ruling biotic and abiotic factors, and the combination of these factors creates a<br />specific microclimate conditions, thus the characteristic fauna. Having applied the<br />molecular analysis with the following species &ndash; Calathus fuscipes, Poecilus versicolor,<br />Amara aenea, Calathus ambiguus, Anchomenus dorsalis, Dolichus halensis,<br />Pterostichus cupreus, Harpalus distinguendus, Harpalus rufipes, Laemostenus<br />terricola, Pterostichus melanarius, Harpalus dimidiatus, Anisodactylus binotatus,<br />Trechus quadristriatus, Carabus coriaceus, Carabus cancellatus &ndash; the BLAST analysis<br />confirmed the status of a species that is primarily identified based on morphological<br />characteristics. Species, in which a high rate of similarity with other species of the same<br />order was found, and for which there is no reference sequence in the gene bank, such as<br />Pterostichus (Poecilus) sericeus, Harpalus azureus, Pterostichus incommodus,<br />Harpalus griseus, Pterostichus (Cophosus) cylindricus, Pterostichus vernalis,<br />Pterostichus (Feronidius) melas i Calosoma auropunctatum represent a contribution to<br />the world bar-coding database.</p>
30

Aspectos microbiológicos e ambientais de candidemias em hospital terciário (HC/FMB/UNESP/Botucatu) localizado na região centro-sul do estado de São Paulo, Brasil

Giacobino, Juliana. January 2018 (has links)
Orientador: Eduardo Luiz Bagagli / Resumo: Infecções fúngicas causadas por leveduras constituem um dos maiores problemas em pacientes hospitalizados em todo o mundo e têm se tornado uma importante causa de morbidade e mortalidade. Embora Candida albicans seja a espécie mais frequentemente isolada e sua forma de infecção ocorra geralmente por translocação endógena, tem-se observado um aumento das espécies não-albicans, com destaque para o complexo C. parapsilosis, cuja principal forma de infecção é exógena, provavelmente pelas mãos dos profissionais de saúde. Este trabalho propôs estudar os aspectos microbiológicos e ambientais das candidemias, com especial atenção para o complexo Candida parapsilosis, no hospital terciário HC/FMB/UNESP, Botucatu, utilizando-se de métodos moleculares, busca ativa dos agentes no ambiente hospitalar (ar, superfícies e mãos de profissionais de saúde), bem como determinar os fatores de virulência e susceptibilidade aos antifúngicos destas espécies e associação com o desfecho clínico. Os isolados clínicos (obtidos de hemoculturas, período 2007-2015) e ambientais (período 2014-2015) de C. parapsilosis sensu lato foram identificados pelo meio CHROMagar Candida, Vitek-2, sequenciamento do rDNA e perfis dos inteins VMA e ThrRS. Fatores de virulência (produção de proteinase, fosfolipase e biofilme) e perfis de susceptibilidade aos antifúngicos anfotericina B, fluconazol, voriconazol, caspofungina e micafungina foram estimados, e dados clínicos obtidos junto aos prontuários dos pacientes. Dentre ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Fungal infections caused by yeasts are serious problems in hospitalized patients around the globe and have become a major cause of morbidity and mortality. Although Candida albicans is the most frequently isolated species and its infection usually occurs by endogenous translocation, an increase of non-albicans species has been observed, with emphasys for the C. parapsilosis complex, whose main route of infection is exogenous, probably by the hands of health professionals. This work proposes to study the microbiological and environmental aspects of candidemia, with special attention to the C. parapsilosis complex, in a public tertiary hospital HC/FMB/UNESP, in Botucatu, using the molecular methods, active search of agents in the hospital environment (air, surfaces and hands of health professionals), as well as to determine the virulence factors and susceptibility to the antifungals of these species and correlate with the patient clinical outcomes. The clinical isolates (obtained of the blood cultures, 2007-2015 period) and environmental (2014-2015 period) of the C. parapsilosis sensu lato were identified by CHROMagar Candida medium, Vitek-2, rDNA sequencing and VMA and ThrRS inteins profiles. Virulence factors (production of proteinase, phospholypase and biofilm) and profiles of susceptibility to antifungals amphotericin B, fluconazole, voriconazole, caspofungin and micafungin were estimated, and the clinical data were obtained from patient’s records. Among the clinical isolat... (Complete abstract click electronic access below) / Doutor

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