No description available.

No description available.

Hydrolytic enzymes such as chitinases and glucanases are implicated in plant defense responses against fungal pathogens. These enzymes are responsible for the breakdown of chitin and glucan, two major components of the fungal cell walls. Genes encoding these enzymes have been used to genetically engineer plants to enhance their protection against fungal pathogens. Western Australia has over 4000 endemic plant species and a largely unknown fungal biota. Given that fungi possessing chitinases and glucanases with novel activities have been isolated in other parts of the world, we propose that fungi from Western Australian soils may possess novel biochemical/enzymatic activities. The aims of this research project were to isolate chitinolytic and glucanolytic fungi from soil and to clone the genes encoding for chitinase and glucanase enzymes. To achieve these aims, fungi with activity against chitin and glucan were isolated, the activity quantified by colorimetric and inhibition assays and gene fragments with homology to known chitinase and glucanase genes were isolated and their sequences determined. Soil fungi were isolated from five locations in and around the Perth Metropolitan area of Western Australia with the use of a medium containing Rose Bengal that eliminates all actinomycetes and most bacteria and reduces the growth of fast growing mold colonies. Forty-one isolates were obtained by this method. Twenty four chitinolytic and glucanolytic fungal isolates were identified by growing them on chitin-containing media to select for those species that utilised chitin/glucan as a carbon source. These were assayed for production of exo- and endochitinolytic and glucanolytic enzymes. Enzyme activity was compared between crude and dialysed supernatants. Exochitinase activity was determined in the supernatants of 4-day old fungal cultures by the release of p-nitrophenol from p-nitrophenyl-N-acetyl-â-D glucosaminide. The supernatants were measured for endochitinase activity determined by the reduction of turbidity of suspensions of colloidal chitin. Glucanase activity was determined by release of reducing sugar (glucose) from laminarin. Supernatants from eleven of the twenty four isolates showed significant levels of enzyme activity. Eleven isolates were assayed for activity against purified cell walls of phytopathogenic fungi. Activity was determined by measuring reducing sugars in the fungal supernatants against cell wall preparations of six economically important plant pathogens. Chitinolytic activity was detected in seven isolates against cell wall preparations of Botrytis cinerea and Rhizoctonia solani, in four isolates against Fusarium solani and Sclerotinia sclerotium; in five isolates against Ascochyta faba and in six isolates against Leptosphaeria maculans. Similarly glucanolytic activity was detected in eight isolates against B. cinerea, in seven against R. solani, in two against F. solani, in three against S. sclerotium and A. faba and in one against L. maculans. The supernatants derived from the isolates were used in a bioassay to determine growth inhibition against live B. cinerea spores by measuring turbidity reduction. Growth inhibition was measured against a control (B. cinerea, grown in medium with no added supernatant). Boiled supernatant did not inhibit the growth of B. cinerea spores but there was 100% inhibition by the crude supernatant from ten of the twenty four isolates. Similarly, supernatants were used to assess growth inhibition against live mycelia cultures of F. solani and S. sclerotium. Growth inhibition of F solani ranged from 9- 59%, boiled and crude supernatants respectively whilst growth inhibition of S. sclerotium ranged from 46-75%, boiled and crude supernatants respectively. Two partial chitinase genes from the soil filamentous ungus Trichoderma asperellum,(ChiA and ChiB) and a novel glucanase gene from the filamentous fungus Aspergillus (Glu1) were cloned. ChiA, was 639 bp long, encoding 191 amino acids with identity to other chitinase genes. Two highly conserved regions, characteristic of glycosyl hydrolases from family 18, were present. ChiB, was 887 bp long and encoded a 293 amino acid sequence that was closely related to an endochitinase gene from the filamentous fungus Trichoderma asperellum. The two highly conserved regions corresponding to the substrate binding and active sites that characterise the glycosyl hydrolases from family 18, also found in ChiA, were found in this gene. Glu1 was 2844 bp long and encoded a 948 amino acid sequence that shared high identity with a â-1, 3-glucanase from the filamentous fungus Aspergillus oryzae. The sequence contained conserved regions found in glycosyl hydrolases from family 17 that encode for substrate binding, N-terminal sequences and putative asparagine linked glycosylation sites. The partial putative sequence ChiA is probably a pseudogene because it has two inframe stop codons. However, once the entire sequence of ChiB is known, both ChiB and the novel glucanase gene Glu1 could be useful contenders for engineering resistance in crop plants.

The master thesis deals with optimization ecological network for biota near Vyskov. First part introduces the current knowledge about the large mammal migration in relation with the territory of Czech Republic, ecological networks, also about TSES. Second part of this project talks about relationship of the Drahanska highland. Third part of this project includes information about my study area. Briefly sum up its history influenced by humans, position of my study area, its abiotic and biotic conditions and landownership relations of area. Further on, it tells something about the wildlife management, mainly about roadkill and introduces the animal occurrence in the area. It includes the updated data about TSES elements inside my study area. Further on examines the current change of supra-regional biocorridor NRBK08 (MH), situation around the town of Vyskov in relation with long distance corridor gets under D1 highway located between Brno and Olomouc city. And introduces information about the main migration obstacles for Drahanska highland. The last part examines mammal migration in the 2014 under the highway bridge. It is also devoted to the optimized ecological network that corresponds with the Long Distance Migration Corridor. The final part talks about optimal size and tree composition in the new planned local biocentre and parts of the supra-regional biocorridor of the ecological network.

No abstract available / Dissertation (MSc)--University of Pretoria, 2013. / am2013 / Genetics / unrestricted

At the macroscopic scale, vertebrate fossils are considered exceptional when non-biomineralized (soft) tissues are preserved. Histologically, high quality is defined by trueness to original shape of a bone, preservation of fine details (e.g. canaliculi), and presence or absence of matrix material in void spaces. Some fossils are hypothesized to preserve cells and durable organelles. Traditionally, cytological details and biomolecular remains have been sought in exceptional fossils. Durable cytological features such as melanosomes do appear to follow feather preservation, but traditionally exceptional fossils are not necessarily exceptional on a microscopic scale. Here, we analyze a feathered dinosaur specimen from the Jehol Lagerstätte to assess claims of blood cell preservation and the state of potential biomolecular preservation. Beipiaosaurus inexpectus is a fairly complete specimen with preserved feathers. Though crushed, fine details in thin section are prevalent. Using Raman spectroscopy, Energy Dispersive X-ray Spectrometry, and Time-of-Flight Secondary Ion Mass Spectroscopy we found no evidence of exceptional molecular preservation. Instead, we found evidence that the vasculature, once hypothesized to contain preserved red blood cells, is filled with clay minerals, with the purported cells chemically indistinguishable from materials of other shapes infilling the vessels. Despite yielding exceptional fossils, the preservational environment of the Jehol biota does not necessarily preserve exceptional details cytologically or biomolecularly. Consequently, we conclude that a systematic approach to biomolecular and cytological preservation studies should rely on traits other than classic exceptional preservation. / Master of Science / What makes a fossil particularly excellent? Traditionally, fossils from animals with skeletons were considered high quality when many or most of the bones from an animal are preserved. If these bones line up with one another like they would in the animal when it was alive (i.e. are articulated) the fossil is even better. To be exceptional, though, soft tissues, or parts of the animal that were not hardened with minerals while the animal lived (e.g. feathers, skin) need to be preserved. All of these traits can be observed with the naked eye. With the use of a microscope, we can see how much a skeleton has been crushed and whether the spaces in the bone for blood vessels and cells have been well preserved. Additionally, we may be able to observe preserved cells, which would be exceptional. On an even smaller scale, the molecules present in a bone might be well or poorly preserved. How much the minerals that make up the bone have changed chemically from when the animal was alive is one indicator of quality. Another might be preservation of molecules that come from the animal such as DNA and the proteins present in bone. In this study, we chose an exceptional fossil based on the traits visible to the naked eye (many of the bones are present and it has feathers) and looked for evidence of cell and unique molecule preservation. On the microscope, we saw beautiful details of the structures in the bone that held bone cells and blood vessels. We also observed red spheres which have been described by other researchers as possible blood cells in the spaces for blood vessels. Using three types of machine which can identify minerals, elements, and molecules in the bone and vessels, we did not find any evidence that the spheres represent preserved blood cells. Nor did we find any evidence of exceptional molecules. However, we did find evidence that the bone itself is not highly changed from when the animal lived, though we see elements and molecules in the vessels that probably did not come from the animal. We started this study knowing that the fossil we chose is exceptional in some ways, but what we found shows that it has a mix of excellent and poor traits visible on the microscope and it does not have any excellent traits in terms of its molecules besides the minerals in the bone itself. We conclude that fossils that are exceptional in the traditional sense are not necessarily exceptional in other ways. / What makes a fossil particularly excellent? Traditionally, fossils from animals with skeletons were considered high quality when many or most of the bones from an animal are preserved. If these bones line up with one another like they would in the animal when it was alive (i.e. are articulated) the fossil is even better. To be exceptional, though, soft tissues, or parts of the animal that were not hardened with minerals while the animal lived (e.g. feathers, skin) need to be preserved. All of these traits can be observed with the naked eye. With the use of a microscope, we can see how much a skeleton has been crushed and whether the spaces in the bone for blood vessels and cells have been well preserved. Additionally, we may be able to observe preserved cells, which would be exceptional. On an even smaller scale, the molecules present in a bone might be well or poorly preserved. How much the minerals that make up the bone have changed chemically from when the animal was alive is one indicator of quality. Another might be preservation of molecules that come from the animal such as DNA and the proteins present in bone. In this study, we chose an exceptional fossil based on the traits visible to the naked eye (many of the bones are present and it has feathers) and looked for evidence of cell and unique molecule preservation. On the microscope, we saw beautiful details of the structures in the bone that held bone cells and blood vessels. We also observed red spheres which have been described by other researchers as possible blood cells in the spaces for blood vessels. Using three types of machine which can identify minerals, elements, and molecules in the bone and vessels, we did not find any evidence that the spheres represent preserved blood cells. Nor did we find any evidence of exceptional molecules. However, we did find evidence that the bone itself is not highly changed from when the animal lived, though we see elements and molecules in the vessels that probably did not come from the animal. We started this study knowing that the fossil we chose is exceptional in some ways, but what we found shows that it has a mix of excellent and poor traits visible on the microscope and it does not have any excellent traits in terms of its molecules besides the minerals in the bone itself. We conclude that fossils that are exceptional in the traditional sense are not necessarily exceptional in other ways.

Este trabalho apresenta estratégias para o ensino do tema biodiversidade na educação básica. O modelo instrucional adotado buscou estimular o educando do ensino fundamental a conhecer a aprender sobre a biodiversidade da sua própria região, além da compreensão de temas relativos ao meio ambiente de forma geral. Para a construção desse modelo foram utilizados, principalmente, os documentos gerados pelo programa Biota-FAPESP. Com isso, procurou-se atender ao item preconizado na Agenda 21 Global, de ensinar em e sobre a sua região. O trabalho foi desenvolvido em três etapas principais: i) aplicação de um questionário para avaliar o nível de conhecimento prévio do estudante sobre o tema; ii) fundamentação teórica; iii) elaboração de cartilhas e almanaques. Ao final do processo, os alunos foram capazes de formular um conceito de biodiversidade, levando em consideração a diversidade das espécies em seus ecossistemas e as relações entre os seres vivos e o ambiente. Pode-se concluir que a metodologia utilizada para a construção do conceito de biodiversidade pelos alunos foi eficiente quando comparada à forma tradicional de ensinar. / In order to develop this work, we made actions in classroom and in visits at field with the objective of the students build the own biodiversity meaning. The work was developed in the three stages: i) application of a questionnaire; ii) theoretical fundaments; iii) activities with primer and almanac. Finally, the students were able to improve their biodiversity concept, considering the diversity of the species, of the ecosystems and of the relationship among living creatures and the environmental, as well, the regional biodiversity through the documents generated by Biota FAPESP program at the end of the 20th century.

Biofiltros em Areia (BFAs) estão sendo aperfeiçoados e testados no Brasil com intuito de auxiliar comunidades carentes que são menos favorecidas em relação à água de boa qualidade. O presente estudo tem como objetivo analisar a qualidade da água tratada pelos BFAs a partir de bioensaios de toxicidade, utilizando larvas do inseto Chironomus sancticaroli, oligoqueto Allonais inaequalis e peixes da espécie Danio rerio, analisando possíveis efeitos tóxicos que possam estar presentes quando a água entra em contato com o PVC do corpo do BFA, além de avaliar a toxicidade da água do poço (AP), captada para abastecimento do BFA. Nos testes de toxicidade agudo (com duração de 96h), crônico (8 dias) e testes mais longos (16 dias) com Chironomus sancticaroli, utilizou-se 240mL da solução-teste (água proveniente do BFA ou AP) em 60g de sedimento controle (areia branca fina para aquário) em 4 réplicas, sendo utilizados 6 larvas de IV ínstar por réplica no teste de toxicidade agudo e de I ínstar nos testes de toxicidade crônico e testes mais longos, alimentadas com 5mL de solução contendo 1000mL de água deionizada e 5,0g de ração tipo Tetramim®. Nos testes de toxicidade agudo e crônico com Allonais inaequalis, foram utilizados 100mL da solução-teste em 5g de sedimento controle em quatro réplicas, sendo utilizados 6 organismos por réplica, alimentados com 5mL de solução contendo 1000mL de água deionizada e 2,0g de ração tipo Tetramim® , com duração de 96h e 10 dias, respectivamente. Foram realizados apenas testes de toxicidade agudo com D. rerio no qual utilizou-se 1000mL de solução-teste e 2 organismos por réplica, em duas réplicas, com duração de 48h, sem alimentação. Os resultados apontam para uma baixa toxicidade do Biofiltro em Areia em relação às três espécies testadas. Além de um índice alto de sobrevivência, a espécie C. sancticaroli concluiu seu ciclo em testes mais longos (16 dias), além de reproduzir-se; a espécie A. inaequalis apresentou 100% de sobrevivência em todos os testes realizados; e não houve mortalidade nos testes de toxicidade agudo com D. rerio. Porém, é importante destacar que estes resultados indicam apenas dados toxicológicos para fauna em relação as efluentes testados, não sendo possível responder a questões de potabilidade da água tratada pelo BFA. Esta dissertação é parte integrante de um projeto financiado pela FAPESP (Processo nº 2014/12712-8), intitulado \"Construção e desempenho de filtros lentos domiciliares conforme a realidade das comunidades isoladas do Brasil\" que esteve sob coordenação da Profa. Dra. Lyda Patricia Sabogal Paz e do Prof. Dr. Juliano José Corbi. / Sand Biofilters (SBFs) are being refined and tested in Brazil. These studies are intended to assist needy communities with difficulties to access drinking water. The aim of the current study is analyze the quality of water treated by SBFs, using toxicity bioassays with larvae of fly Chironomus sancticaroli, oligochaete Allonais inaequalis and zebrafishes Danio rerio, as well as verify the occurrence of possible toxic effects of PVC (Polyvinyl chloride), a material component of SBFs\' structure, and also evaluate the toxicity of well water quality (AP), extracted to supply the SBF. In the acute toxicity tests (lasting 96h), chronic (lasting 8 days) and longer tests (lasting 16 days), all involving Chironomus sancticaroli, it was placed 240mL of sample solution (water from SBF or AP) in 60g of sterile sediment (fine white sand for aquarium) in four replicates, where was added 6 larvae (IV instar) in each replicate for the acute toxicity test, and larvae of I instar for chronic toxicity tests, even as in the longer tests. The organisms were fed by 5mL of Tetramim ® solution (5g/L). In the acute and chronic toxicity tests with Allonais inaequalis, it was applied 100mL of sample solution with 5g of sterile sand in four replicates, as above, it was added 6 organisms by replicate, fed by Tetramim ® (2,0g/L) for both tests, with lasting of 96h and 10 days, respectively. It was performed only tests of acute toxicity for D. rerio in which was added 1000mL of sample solution and 2 organisms for each replicate, with two replicates, with duration of 48h, without feeding. The results showed a low toxicity of sand Biofilters\' effluent in relation to the three species exposed to the liquid. Furthermore, the results demonstrate a high index of survival, even in longer tests (16 days lasting) with C. sancticaroli when the organisms could conclude their life cycle. However, besides the 100% of survival in all test, for the A. inaequalis were also observed reproduction; and in the acute toxicity tests for D. rerio it wasn\'t noticed mortality. Although, it is important to highlight that these results represent only toxicological data concerning to aquatic wildlife exposed to effluents, what is not regarding to potability issues of the water treated by SBF. This dissertation is a constituent part of a project financiad by FAPESP (Process number 2014/12712-8), entitled \"Building and performance of domiciliary slow filters according to the isolated comunities reality in Brazil\" which was coordinated by Professors PhD. Lyda Patricia Sabogal Paz and PhD. Juliano José Corbi.

O continente Antártico é considerado um ambiente pouco impactado, porém com o aumento das atividades humanas e do número de estações de pesquisa, há uma tendência de aumento dos níveis de alguns contaminantes. Neste estudo estabeleceram-se os níveis de arsênio, cádmio, chumbo, cobre, cromo, mercúrio, níquel, zinco, e a razão isotópica de nitrogênio, em sedimentos e na biota, coletados em 2003, na Baía do Almirantado, Ilha Rei George, Antártica. Os metais pesados foram quantificados por Espectrometria de Emissão Ótica com Plasma Indutivamente Acoplado (ICP-OES). Os resultados demonstraram elevadas concentrações de cobre e zinco nos sedimentos, atribuídas à composição geológica da região. Verificou-se bioacumulação de arsênio na biota da Baía do Almirantado, e bioacumulação de cádmio e zinco na biota da Enseada Martel. Somente cobre apresentou tendência de biomagnificação na teia trófica estudada. O molusco Laternula elliptica se apresentou como um bom biomonitor das concentrações de metais pesados para o ambiente Antártico. Além disso, os resultados foram úteis para o conhecimento dos níveis de metais pesados nas condições pré-acidente da Estação Antártica Comandante Ferraz (EACF), que ocorreu em 2012, e para comparação com as atuais condições, dentro dos trabalhos de monitoramento que estão sendo realizados pelo INCT-APA (Instituto Nacional de Ciência e Tecnologia Antártico de Pesquisas Ambientais). / The Antarctic continent is considered a slightly affected environment, but as the human activities and the number of research stations are increasing, there is a tendency of a higher level of some contaminants. This study established the levels of arsenic, cadmium, lead, copper, chromium, mercury, nickel, zinc, and nitrogen isotope ratios in sediments and biota collected in 2003, in Almirantado Bay, Rei George Island, Antarctica. Heavy metals were quantified by Optical Emission Spectrometry with Inductively Coupled Plasma (ICP-OES). The results showed high concentrations of copper and zinc in sediments attributed to the geological composition of the region. There was arsenic bioaccumulation in biota of Almirantado Bay, and bioaccumulation of cadmium and zinc in the biota of Martel Bay. Only copper tended to biomagnification in the trophic web studied. The clam Laternula elliptica presented itself as a good biomonitor of heavy concentrations of metals for Antarctic environment. Moreover, the results were useful for the acknowledge of the levels of heavy metals in pre-accident conditions of Comandante Ferraz Antarctic Station (EACF), which occurred in 2012, and for comparison with current conditions within the monitoring work now performed by INCT-APA (National Antartic Institute of Science and Technology Environmental Research).

O continente Antártico é considerado um ambiente pouco impactado, porém com o aumento das atividades humanas e do número de estações de pesquisa, há uma tendência de aumento dos níveis de alguns contaminantes. Neste estudo estabeleceram-se os níveis de arsênio, cádmio, chumbo, cobre, cromo, mercúrio, níquel, zinco, e a razão isotópica de nitrogênio, em sedimentos e na biota, coletados em 2003, na Baía do Almirantado, Ilha Rei George, Antártica. Os metais pesados foram quantificados por Espectrometria de Emissão Ótica com Plasma Indutivamente Acoplado (ICP-OES). Os resultados demonstraram elevadas concentrações de cobre e zinco nos sedimentos, atribuídas à composição geológica da região. Verificou-se bioacumulação de arsênio na biota da Baía do Almirantado, e bioacumulação de cádmio e zinco na biota da Enseada Martel. Somente cobre apresentou tendência de biomagnificação na teia trófica estudada. O molusco Laternula elliptica se apresentou como um bom biomonitor das concentrações de metais pesados para o ambiente Antártico. Além disso, os resultados foram úteis para o conhecimento dos níveis de metais pesados nas condições pré-acidente da Estação Antártica Comandante Ferraz (EACF), que ocorreu em 2012, e para comparação com as atuais condições, dentro dos trabalhos de monitoramento que estão sendo realizados pelo INCT-APA (Instituto Nacional de Ciência e Tecnologia Antártico de Pesquisas Ambientais). / The Antarctic continent is considered a slightly affected environment, but as the human activities and the number of research stations are increasing, there is a tendency of a higher level of some contaminants. This study established the levels of arsenic, cadmium, lead, copper, chromium, mercury, nickel, zinc, and nitrogen isotope ratios in sediments and biota collected in 2003, in Almirantado Bay, Rei George Island, Antarctica. Heavy metals were quantified by Optical Emission Spectrometry with Inductively Coupled Plasma (ICP-OES). The results showed high concentrations of copper and zinc in sediments attributed to the geological composition of the region. There was arsenic bioaccumulation in biota of Almirantado Bay, and bioaccumulation of cadmium and zinc in the biota of Martel Bay. Only copper tended to biomagnification in the trophic web studied. The clam Laternula elliptica presented itself as a good biomonitor of heavy concentrations of metals for Antarctic environment. Moreover, the results were useful for the acknowledge of the levels of heavy metals in pre-accident conditions of Comandante Ferraz Antarctic Station (EACF), which occurred in 2012, and for comparison with current conditions within the monitoring work now performed by INCT-APA (National Antartic Institute of Science and Technology Environmental Research).