L’objectif de ce travail est d’évaluer les risques chimiques et microbiologiques de la chaîne alimentaire des larves de dorade royale en milieu aquacole, nous avons déterminé les concentrations létales (LC50) du Cadmium (Cd), du Cuivre (Cu) et du Nickel (Ni) chez le copépode calanoïde Eurytemora affinis. Nous avons ensuite quantifié, par voie d’entrée dissoute de ces métaux seuls et en mélange, la bioaccumulation dans ces organismes. Le copépode E. affinis accumule le Cd, le Cu et le Ni et par conséquent pourraient être utilisé comme indicateur biologique dans des études de biosurveillance pour l'évaluation des risques environnementaux. L’étude de la diversité microbienne de la chaine trophique (l’eau de mer, les microalgues et les copépodes) des larves de dorade a conduit à l’isolement de 434 isolats bactériens dont 246 ont été positivement identifiés par MALDI-TOF-MS. Parmi les espèces identifiées, Vibrio anguillarum, V. alginolyticus, Bacillus cereus, Pseudomanas anguilliseptica, P. putida, Staphylococcus epidermidis et S. hominis sont des pathogènes des poissons. Cependant, cinq souches non pathogènes de B. pumilus (nommées 18 COPS, 35A COPS, 35R COPS, 38 COPS et 40A COPS) montrent une activité antagoniste contre certaines souches pathogènes précédemment citées. La souche B. pumilus 35R COPS produit deux classes de composés antimicrobiens de type peptides non-ribosomaux : des surfactines mais aussi les amicoumacines A et B et les phosphoamicoumacines A et B identifiées et caractérisées par MALDI-TOF-MS et HPLC-MS/MS. La souche B. pumilus 38 COPS, quant-à-elle, produit également des membres de la famille des surfactines mais surtout une bactériocine appartenant à la famille de la circularine A/uberolysine. L’analyse in silico, des génomes de ces souches révèle la présence de clusters de gènes codant pour les surfactines et les amicoumacines et un gène ribosomique codant une bactériocine appartenant à la famille de la circularine A/uberolysine. / This work aimed at evaluating the chemical and microbiological risks associated with the larval food chain from seabream. Therefore, we have determined the lethal concentrations (LC50) of Cadmium (Cd), Copper (Cu) and Nickel (Ni) in Eurytemora affinis calanoid copepods. Then, we then quantified, through the dissolved entry way of these metals either alone or in combination. The E. affinis copepods appeared to accumulate Cd, Cu and Ni and therefof could be used in the bio-monitoring studies for assessing the environmental risks. The microbial diversity study undertaken on the trophic chain (seawater, microalgae and copepods) of seabream larvae permitted isolation of 434 bacterial isolates, and 246 isolates were positively identified by MALDI-TOF-MS technology. The bacterial species identified included notably Vibrio anguillarum, V. alginolyticus, Bacillus cereus Pseudomanas anguilliseptica, P. putida, Staphylococcus epidermidis and S. hominis, which are considered as fish pathogens. On the other hand, five non-pathogenic strains included Bacillus pumilus as strains 18 COPS, 35A COPS, 35R COPS, 38 COPS and 40A COPS, which displayed antagonistic activities against towards some pathogenic species. To be noted that antagonistic strain B. pumilus strain 35R COPS is able to produce two non-ribosomal antimicrobial peptides (NRPS), which are: surfactins and amicoumacins A and B and phosphoamicoumacins A and B based on MALDI-TOF-MS and HPLC-MS/MS data. Moreover, B. pumilus 38 COPS was characetrized as antagonistic strain producing neverthemless, a surfactine (NRPS family) and concomitantly a bacteriocin belonging to the circularin A/uberolysin family. The in silico analyses of their genomes revealed the presence of gene clusters coding for the surfactin- and amicoumacin-families and a ribosomal gene encoding a bacteriocin belonging to the circularin A/uberolysin bacteriocin family.

Made available in DSpace on 2015-09-17T15:24:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-21. Added 1 bitstream(s) on 2015-09-17T15:48:06Z : No. of bitstreams: 1 000845905.pdf: 877611 bytes, checksum: c5c230ba41c8a163a8d990590eb9f720 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Os surfactantes são moléculas que por apresentarem porções hidrofílicas e hidrofóbicas são capazes de reduzir a tensão superficial de um meio contendo água. Esta característica é importante para aplicações industriais fazendo destes tenso-ativos compostos de grande interesse. Por outro lado, os biossurfactantes são moléculas que apresentam características surfactantes e são produzidas por microrganismos. Apresentam grande poder tenso-ativo e a vantagem de poderem ser produzidos a partir de substratos renováveis; além de serem biodegradáveis, apresentarem baixa toxicidade e especificidade química. O objetivo deste trabalho foi verificar a produção de um biossurfactante pela bactéria Bacillus pumilus utilizando manipueira, melaço, soro de queijo e sacarose como substratos. Verificar a produção e propriedades deste biossurfactante em escala ampliada (2L) utilizando sacarose como fonte de carbono, bem como, seu potencial de aplicação em biorremediação de um solo agrícola contaminado com hidrocarbonetos. Todos os experimentos foram planejados utilizando a metodologia de superfície de resposta. Para todos os experimentos de fermentação foram realizadas as análises de índice de emulsificação, tensão superficial e crescimento microbiano. Em todos os substratos testados foi determinada a concentração bruta e a estabilidade do biossurfactante produzido frente às condições extremas de temperatura, pH e salinidade. Os resultados obtidos mostraram que a bactéria Bacillus pumilus foi capaz de crescer e produzir biossurfactante com todos os substratos testados, reduzindo a tensão superficial em até 46; 37; 47 e 32 mNm-1com manipueira, melaço, soro de queijo e sacarose, respectivamente. O biossurfactante foi capaz de apresentar índices de emulsificação de até 50% com tolueno e óleo de soja. A produção bruta obtida foi (g/L): 1,12; 0,35, 1,4 e 5,1 para manipueira,... / Surfactants are molecules that contain hydrophilic and hydrophobic moieties, and because of this, they are capable of reducing the surface tension of a medium containing water. This feature is important for industrial applications by making these surface active compounds of high interest. On the other hand, biosurfactants are molecules that exhibit surfactant characteristics and are produced by microorganisms. They have large surface- active power and the advantage of being produced from renewable substrates; in addition to being biodegradable, have low toxicity and chemical specificity. The objective of the work was to investigate the production of a biosurfactant by Bacillus pumilus using cassava, molasses, cheese whey and sucrose as substrates; check the production and properties of this biosurfactant on a large scale (2L) using sucrose as carbon source, as well, as their potential for application in bioremediation of an agricultural soil contaminated by hydrocarbons. All experiments were planned using surface response methodology. For all fermentation experiments the emulsification index, surface tension, and microbial growth analyses were performed. In all substrates tested, the crude concentration and the stability of the biosurfactant produced were determined in extremes conditions of temperature, pH and salinity. The results showed that the bacterium Bacillus pumilus was able to grow and produce biosurfactant with all substrates tested, reducing the surface tension to 46, 37, 47 and 32 mNm-1 with cassava, molasses, whey and sucrose, respectively. The biosurfactant was able to present emulsification rates of up to 50% with toluene and soybean oil. The crude production obtained was (g/L): 1.12; 0.35, 1.4 and 5.1 for cassava, molasses, whey and sucrose, respectively. The biopolymer produced was stable across variations in temperature, pH and salinity. In the large-scale fermentation with sucrose ...

Submitted by Alisson Mota (alisson.davidbeckam@gmail.com) on 2015-07-06T19:29:41Z No. of bitstreams: 1 Dissertação - Jefferson Ferreira dos Santos.pdf: 12615415 bytes, checksum: 28e029e2822cb4b7e87ef5b2dc18919b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-06T19:38:38Z (GMT) No. of bitstreams: 1 Dissertação - Jefferson Ferreira dos Santos.pdf: 12615415 bytes, checksum: 28e029e2822cb4b7e87ef5b2dc18919b (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-06T19:42:38Z (GMT) No. of bitstreams: 1 Dissertação - Jefferson Ferreira dos Santos.pdf: 12615415 bytes, checksum: 28e029e2822cb4b7e87ef5b2dc18919b (MD5) / Made available in DSpace on 2015-07-06T19:42:38Z (GMT). No. of bitstreams: 1 Dissertação - Jefferson Ferreira dos Santos.pdf: 12615415 bytes, checksum: 28e029e2822cb4b7e87ef5b2dc18919b (MD5) Previous issue date: 2014-04-11 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / The ever increasing consumption of fossil fuels puts oil as one of the most important products of human consumption. Undeniable technological and even social breakthroughs are coming era of "post-oil" . But this unbridled consumption has led to serious environmental aquatic ecosystems and land damage. Viable alternatives that promote the recovery of areas affected by these compounds are undoubtedly of utmost importance given the current scenario. Bioremediation has proven very promising in this area . And increasingly, new technology are aggregated to this field of research in an attempt to obtain more conclusive results, thus improving the overall understanding of how microorganisms interact in promoting bioremediation. This is precisely what has been proposed in this work. The use of Mass Spectrometry (MS) for identification of proteins of bacteria capable of growth in xenobiotics. Initially was selected near the Refinaria Isaac Sabbá (REMAN) species B. pumilus, with the potential ability to grow in petroleum and diesel. After conducting tests using oil and diesel separately with unique carbon sources, was proven capacity for growth of the species in these xenobiotics. Better growth/biomass production was recorded in crops having diesel as finte carbon, as well as the best protein yield. The proteomic profile in petroleum, diesel and yeast extract was determined. Comparisons between these profiles showed very marked difference between them. For analysis via mass spectrometry were selected 96 spots of B. pumilus were selected and identified by Peptide Mass Fingerprint (PMF) . Of this total 54 protein spots were identified. Some with potential involvement in direct bioremediation processes by B. pumilus. However, further studies are necessary to effective proof of proteins in the pathways of degradation of xenobiotics. / O consumo cada vez crescente dos combustíveis fósseis coloca o petróleo como um dos mais importantes produtos do consumo humano. Inegáveis avanços tecnológicos e até mesmo sociais são advindos da era “Pós-Petróleo”. Porém, esse consumo desenfreado tem gerado graves danos ambientais aos ecossistemas aquáticos e também terrestres. Alternativas viáveis capazes de promover a recuperação de áreas afetadas por esses compostos são sem dúvida de extrema importância diante do cenário atual. A biorremediação tem se mostrado bastante promissora nessa área. E cada vez mais novas tecnologia são agregadas a esse campo de investigação na tentativa de obter resultados mais conclusivos, melhorado assim, o entendimento global de como os microorganismos interagem na promoção da biorremediação. É justamente isso o que foi proposto nesse trabalho. O emprego da espectrometria (MS) de massa para identificação de proteínas de bactéria com capacidade de crescimento em xenobióticos. Inicialmente foi selecionada nas proximidades da Refinaria Isaac Sabbá (REMAN) a espécie B. pumilus, com potencial capacidade de crescer em petróleo e diesel. Após a realização de testes usando petróleo e diesel, separadamente, com fontes únicas de carbono, foi comprovada a capacidade de crescimento da espécie nesses xenobióticos. Melhor crescimento/produção de biomassa foi registrado nos cultivos tendo o diesel como finte de carbono, bem como o melhor rendimento protéico. O perfil proteômico em petróleo, diesel e extrato de levedura foi determinado. As comparações entre tais perfis demonstrou diferenças bastante acentuadas entre eles. Para análise via espectrometria massas foram selecionados 96 spots de B. pumilus e identificados por Peptide Mass Fingerprint (PMF). Desse total de spots, 54 proteínas foram identificadas. Algumas delas com potencial envolvimento nos processos diretos de biorremediação apresentada por B. pumilus. Porém, mais estudos são necessário para efetiva comprovação dessas proteínas nas vias de degradação de xenobióticos.

Agarwood mainly formed by Aquilaria species is an economically and pharmaceutically important natural product used for the production of incense, perfumes and traditional medicines in Asia. Endophytic bacteria are potentially important in producing pharmaceutical compounds found in the plants. The aim of this research is to isolate, classify and identify major endophytic bacteria groups associated with agarwood of Aquilaria crassna species in Khanh Hoa province, Vietnam. Agarwood samples were collected and surface-sterilized, and total endophytic bacteria were isolated on Tryptic Soy Agar by the spread plate method. Major bacterial groups were classified according to the Bergey’s system. The 16S rRNA gene fragments were amplified using PCR method, and bacterial isolates were identified using this gene sequence similarity based method. The results showed that from 0.121 g of agarwood, total 26 bacterial isolates were purified and divided into 7 separated groups, in which the group II of Gram-positive spore-forming bacteria was the most dominant. Especially, two dominant strains, T14 of group II, and T15 of group VII, were identified as Bacillus pumilus and Alcaligenes faecalis, respectively.!To our knowledge, it is the first time that biodiversity of bacterial endophytes associated with agarwood from Aquilaria crassna in Vietnam has been reported, which requires of further study to understand the relationship of endophytic bacteria to agarwood-producing Aquilaria crassna species as well as explore their potential applications towards the development of valuable bioactive compounds. / Trầm hương, chủ yếu được tạo ra từ các loài cây Dó (Aquilaria), là một sản phẩm tự nhiên có giá trị kinh tế và y học đã được sử dụng để sản xuất hương, nước hoa và các dược phẩm truyền thống ở châu Á. Vi khuẩn nội cộng sinh thực vật được cho là một nguồn quan trọng cho các dược phẩm có nguồn gốc thực vật. Mục tiêu của nghiên cứu này là nhằm phân lập, phân loại và định danh các nhóm vi khuẩn chính trên Trầm hương Khánh Hòa, Việt Nam. Các mẫu Trầm hương được thu nhận và vô trùng bề mặt dùng để phân lập vi khuẩn tổng số trên môi trường TSA bằng phương pháp trải đĩa. Các nhóm vi khuẩn chính được phân loại dựa theo hệ thống chuẩn Bergey. Đoạn gen mã hóa 16S rRNA được khuếch đại bằng phương pháp PCR, và các chủng vi khuẩn được định danh bằng phép so sánh độ tương đồng trình tự của đoạn gen này. Kết quả cho thấy từ 0,121 g mẫu trầm hương, chúng tôi đã phân lập được 26 chủng vi khuẩn và phân chúng vào 7 nhóm chính, trong đó nhóm II bao gồm các vi khuẩn Gram dương sinh bào tử là nhóm chiếm ưu thế nhất. Đặc biệt, có 2 chủng ưu thế là chủng T14 thuộc nhóm II và chủng T15 thuộc nhóm VII đã được định danh tương ứng là Bacillus pumilus và Alcaligenes faecalis.!Đây là nghiên cứu đầu tiên về đa dạng sinh học của các nhóm vi khuẩn chính trên Trầm hương Khánh Hòa. Vì vậy, cần có những nghiên cứu tiếp theo nhằm tìm hiểu mối quan hệ giữa các vi khuẩn nội cộng sinh với cây Dó bầu (Aquilaria crassna) tạo trầm cũng như khai thác những ứng dụng tiềm năng của các vi khuẩn này theo hướng phát triển các hoạt chất sinh học có giá trị.

Der Einfluss der Rhizosphärenbakterien Bacillus subtilis FZB 24, FZB 37, FZB 42, Bacillus pumilus RK 13 und Streptomyces graminofaciens und des Algenpräparates Goemar Fruton Spezial® (Ascophyllum nodosum) auf Wachstum, Ertrag und Toleranz gegenüber Fusarium spp. bei Spargel wurde untersucht. Es wurden mehrjährige Parzellenfeldversuche und Modellversuche unter kontrollierten Bedingungen mit natürlich Fusarium spp. belasteten Böden und mit speziellen Fusarium Erregern inokulierten Substraten durchgeführt. Die Analyse des verwendeten Nachbaubodens ergab eine wesentlich höhere Kontamination mit Fusarium spp. (56%) in Relation zum Fruchtfolgeboden (14%). F. oxysporum war die dominante Fusariumart. Desweiteren wurde eine ganze Reihe anderer Arten, wie z.B. F. acuminatum, F. culmorum, F. proliferatum und F. culmorum bestimmt, was die Komplexität der Wurzel- und Stängelfäuleerkrankung belegt. Analog zu diesem Ergebnis konnten im Vergleich zur Fruchtfolgevariante von den in dem Nachbauboden kultivierten Spargelpflanzen signifikant mehr Fusarium spp. (80%) isoliert werden. Das Wurzelwachstum war hier extrem reduziert. Unter diesen konduktiven Bedingungen gelang es durch Rhizombakterisierung (107 cfu/ml) mit B. subtilis FZB 42, eine signifikante Förderung des Wurzelwachstums relativ zur unbehandelten Kontrolle und zur B. subtilis FZB 37 Variante zu erzielen. In Pathogenitätstests konnten F. culmorum, F. oxysporum und F. proliferatum als bedeutende Spargelpathogene mit signifikant reduzierter Trieb- und Wurzelmasse nachgewiesen werden, während F. acuminatum nur geringe Symptome verursachte. Eine bakterielle Saatgutbeizung (108 cfu/ml) und zusätzliche präinfektionelle Gießapplikation (107 cfu/ml) konnte eine Infektion mit F. oxysporum f.sp. asparagi nicht verhindern. Dennoch waren die negativen Effekte im Vergleich mit der nicht bakterisierten Kontrolle in den B. subtilis Varianten FZB 24 und vor allem bei FZB 42 deutlich kompensiert. Diese Ergebnisse lassen vermuten, dass eine Resistenz- bzw. Toleranzinduktion ein potentieller Wirkmechanismus der Bakterien ist. In den Feldversuchen erbrachte eine Bakterisierung einjähriger Rhizome (107 cfu/ml) zur Pflanzung und ergänzende Gießbehandlungen (108 cfu/ml/1l/m) in den folgenden 2 Jahren eine tendenzielle Reduzierung der Trieblänge und des Triebdurchmessers, insbesondere in Kombination mit dem Algenpräparat. Einen eindeutigen Einfluss auf das Sortierungsergebnis konnte nicht nachgewiesen werden. Im Gegensatz hierzu resultierte eine Saatgutbakterisierung mit B. subtilis FZB 24 in einem generell geförderten Triebwachstum und einer signifikant gesteigerten Wurzelentwicklung. Auch die anderen Mikroorganismen erbrachten eine tendenzielle Wuchsförderung. Es konnte eine gesicherte Korrelation zwischen Wurzelmasse und Knospenanzahl ermittelt werden. Blattapplikationen mit Goemar Fruton Spezial® während der Hauptvegetationsphase hatten keinen Einfluss auf das Pflanzenwachstum. / The effects of the rhizobacteria Bacillus subtilis, Strain FZB 24, FZB 37and FZB 42, Bacillus pumilus RK 13, Streptomyces graminofaciens N6 and the alga Ascophyllum nodosum (Goemar Fruton Spezial®) on plant growth, yield and Fusarium spp. tolerance of Asparagus officinalis (L.) were investigated. The trials were carried out under field conditions over sev-eral years by sawing and planting and as pot trials in the greenhouse and climate chamber with soil naturally infested with Fusarium spp. and with steamed substrates inoculated with particularly Fusarium species. Asparagus replant soil was considerable higher infested with Fusarium spp. (56 %) than fresh soil without asparagus history (14 %). F. oxysporum was the most determined Fusarium species followed by F. redolens, F. acuminatum, F. culmorum, F. proliferatum, F. solani a.o.. That indicates the complex character of root and crown rot of asparagus. From asparagus plants cultivated in replant soil a significant higher rate of Fusarium isolations (80 %) could be worked out compared to the fresh soil treatment. The root growth in the contaminated soil was extremely reduced. In these conductive situation bacterial treatments with B. subtilis FZB 42 done as rhizom soaking (107cfu/ml) resulted in significantly increased root growth up to 32, 9 % related to the non treated control. B. subtilis FZB 37 was ineffective. F. oxysporum, F. culmorum and F. proliferatum showed a high pathogenicity to asparagus seedlings, manifested in significantly reduced fresh and dry weights of shoots and roots, while F. acuminatum caused only slight symptoms. None of the tested microorganisms applicated as seed coating (108 cfu/ml) and substrate drenching before pathogen inoculation (107 cfu/ml) could prevent seedlings from being infected by F. oxysporum f. sp. asparagi. The negative effects of infection are significantly compensated by Bacillus subtilis FZB 42 and FZB 24. The results support the conclusion, that induced tolerance is a potential mechanism of bacterial mode of action. Under field conditions rhizom bacterization of one year old asparagus plants (107 cfu/ml) before planting and additional soil drenching (108 cfu/ml/1,5l/m) during the following two years reduced shoot length, shoot diameter and yield, especially in combination with Goemar Fruton Spezial®. There was no remarkable influence on spear quality. Seed coating with B. subtilis FZB 24 (108 cfu/ml) and a soil drenching (108 cfu/ml/1l/m) in summer however re-sulted in higher shoots and significantly increased fresh weight and dry substance of roots. But also the other tested microorganisms showed a plant growth promoting trend. There was a significant positive correlation between root fresh weight and number of buds. By spraying the phylloclads four times during the main vegetation season the alga application only gave a slight improvement of plant growth in the field trials.

Bt é uma bactéria formadora da proteína Cry1Ac, tóxica a lepidópteros. Plantas geneticamente modificadas expressam essa toxina. O objetivo deste trabalho foi isolamento e identificação de bactérias do solo de algodão transgênico e convencional potencialmente biodegradadoras dessa proteína. Estudou-se a dinâmica de crescimento das bactérias em meios com a proteína Cry1Ac ou glucose, a biodegradação, os genes apr, npr e sub. Em solo de algodão convencional a contagem foi menor; para algodão transgênico foi maior. Seis bactérias foram identificadas. O crescimento da bactéria B. pumilus foi maior em meio com a proteína Cry1Ac que glucose e a bactéria G. rubripertincta em meio contendo glucose teve o crescimento maior do que com a proteína Cry1Ac. Foi verificada a biodegradação da proteína pelas bactérias Bp e Gr. Os genes apr e sub foram detectados em solo de planta transgênica e o gene npr não. / Bacillus thuringiensis is a bacterium that forms the Cry1Ac protein, toxic to lepidopteran. Genetically modify plants expressing this toxin. The objective of this work was the isolation and identification bacteria from soil cultivated with transgenic cotton and conventional with potential degradation of this protein. We studied the dynamics growth from bacteria in media with the Cry1Ac protein or glucose, the biodegradation genes apr, npr and sub. In soil of conventional cotton, count was lower, for Bt cotton were higher. Six strains were identified. The growth bacterium B. pumilus was higher in medium with the Cry1Ac protein than bacteria with glucose and G. rubripertincta in medium containing glucose had higher growth than the Cry1Ac protein. Were verified Cry1Ac protein degradation by the bacteria Bp and Gr. Genes apr and sub were detected in soil with transgenic plant and the gene npr were not.

Bt é uma bactéria formadora da proteína Cry1Ac, tóxica a lepidópteros. Plantas geneticamente modificadas expressam essa toxina. O objetivo deste trabalho foi isolamento e identificação de bactérias do solo de algodão transgênico e convencional potencialmente biodegradadoras dessa proteína. Estudou-se a dinâmica de crescimento das bactérias em meios com a proteína Cry1Ac ou glucose, a biodegradação, os genes apr, npr e sub. Em solo de algodão convencional a contagem foi menor; para algodão transgênico foi maior. Seis bactérias foram identificadas. O crescimento da bactéria B. pumilus foi maior em meio com a proteína Cry1Ac que glucose e a bactéria G. rubripertincta em meio contendo glucose teve o crescimento maior do que com a proteína Cry1Ac. Foi verificada a biodegradação da proteína pelas bactérias Bp e Gr. Os genes apr e sub foram detectados em solo de planta transgênica e o gene npr não. / Bacillus thuringiensis is a bacterium that forms the Cry1Ac protein, toxic to lepidopteran. Genetically modify plants expressing this toxin. The objective of this work was the isolation and identification bacteria from soil cultivated with transgenic cotton and conventional with potential degradation of this protein. We studied the dynamics growth from bacteria in media with the Cry1Ac protein or glucose, the biodegradation genes apr, npr and sub. In soil of conventional cotton, count was lower, for Bt cotton were higher. Six strains were identified. The growth bacterium B. pumilus was higher in medium with the Cry1Ac protein than bacteria with glucose and G. rubripertincta in medium containing glucose had higher growth than the Cry1Ac protein. Were verified Cry1Ac protein degradation by the bacteria Bp and Gr. Genes apr and sub were detected in soil with transgenic plant and the gene npr were not.

Agarwood mainly formed by Aquilaria species is an economically and pharmaceutically important natural product used for the production of incense, perfumes and traditional medicines in Asia. Endophytic bacteria are potentially important in producing pharmaceutical compounds found in the plants. The aim of this research is to isolate, classify and identify major endophytic bacteria groups associated with agarwood of Aquilaria crassna species in Khanh Hoa province, Vietnam. Agarwood samples were collected and surface-sterilized, and total endophytic bacteria were isolated on Tryptic Soy Agar by the spread plate method. Major bacterial groups were classified according to the Bergey’s system. The 16S rRNA gene fragments were amplified using PCR method, and bacterial isolates were identified using this gene sequence similarity based method. The results showed that from 0.121 g of agarwood, total 26 bacterial isolates were purified and divided into 7 separated groups, in which the group II of Gram-positive spore-forming bacteria was the most dominant. Especially, two dominant strains, T14 of group II, and T15 of group VII, were identified as Bacillus pumilus and Alcaligenes faecalis, respectively.!To our knowledge, it is the first time that biodiversity of bacterial endophytes associated with agarwood from Aquilaria crassna in Vietnam has been reported, which requires of further study to understand the relationship of endophytic bacteria to agarwood-producing Aquilaria crassna species as well as explore their potential applications towards the development of valuable bioactive compounds. / Trầm hương, chủ yếu được tạo ra từ các loài cây Dó (Aquilaria), là một sản phẩm tự nhiên có giá trị kinh tế và y học đã được sử dụng để sản xuất hương, nước hoa và các dược phẩm truyền thống ở châu Á. Vi khuẩn nội cộng sinh thực vật được cho là một nguồn quan trọng cho các dược phẩm có nguồn gốc thực vật. Mục tiêu của nghiên cứu này là nhằm phân lập, phân loại và định danh các nhóm vi khuẩn chính trên Trầm hương Khánh Hòa, Việt Nam. Các mẫu Trầm hương được thu nhận và vô trùng bề mặt dùng để phân lập vi khuẩn tổng số trên môi trường TSA bằng phương pháp trải đĩa. Các nhóm vi khuẩn chính được phân loại dựa theo hệ thống chuẩn Bergey. Đoạn gen mã hóa 16S rRNA được khuếch đại bằng phương pháp PCR, và các chủng vi khuẩn được định danh bằng phép so sánh độ tương đồng trình tự của đoạn gen này. Kết quả cho thấy từ 0,121 g mẫu trầm hương, chúng tôi đã phân lập được 26 chủng vi khuẩn và phân chúng vào 7 nhóm chính, trong đó nhóm II bao gồm các vi khuẩn Gram dương sinh bào tử là nhóm chiếm ưu thế nhất. Đặc biệt, có 2 chủng ưu thế là chủng T14 thuộc nhóm II và chủng T15 thuộc nhóm VII đã được định danh tương ứng là Bacillus pumilus và Alcaligenes faecalis.!Đây là nghiên cứu đầu tiên về đa dạng sinh học của các nhóm vi khuẩn chính trên Trầm hương Khánh Hòa. Vì vậy, cần có những nghiên cứu tiếp theo nhằm tìm hiểu mối quan hệ giữa các vi khuẩn nội cộng sinh với cây Dó bầu (Aquilaria crassna) tạo trầm cũng như khai thác những ứng dụng tiềm năng của các vi khuẩn này theo hướng phát triển các hoạt chất sinh học có giá trị.

O cianeto é um composto tóxico, que pode ser encontrado no ambiente de maneira natural ou como resultado de atividades antropogênicas tais como a mineração de ouro e a indústria da galvanoplastia. Dentre as diferentes espécies do composto, o cianeto de hidrogênio (HCN) é considerado o mais tóxico, mesmo concentrações de 100ppm são letais para os seres humanos. Para a degradação destes compostos de cianeto a compostos menos tóxicos existem diferentes métodos de tratamento que podem ser químicos, físicos ou biológicos. O presente trabalho estudou a capacidade de degradação bacteriana de cianeto com cepas nativas isoladas de rejeito de mineração de ouro. Para isto, foram escolhidas três cepas dentro de um grupo de vinte cepas isoladas previamente. As cepas foram identificadas mediante as técnicas de MALDI-TOF e sequenciamento do gene 16s. Posteriormente realizou-se o processo de ativação e crescimento bacteriano no qual foram determinados os parâmetros de crescimento para cada uma das cepas, tais como pH, agitação, temperatura e meio de cultura. Após a etapa de ativação bacteriana, realizou-se a adaptação das três cepas em ambientes alcalinos. Nesta etapa, foram feitos ensaios em frascos agitados e avaliou-se o crescimento celular em função da formação de células viáveis para diferentes condições de pH (7; 8; 9,10 e 11). Com as cepas adaptadas ao pH 10 foram realizados ensaios de degradação bacteriana de cianeto em frascos agitados contendo 100mL de solução sintética de cianeto de potássio e 0,2mL de inoculo bacteriano. A concentração da solução de cianeto foi de 500mgL-1 e o pH de 10. Foram avaliadas três condições de temperatura (37, 32 e 27oC). Durante os ensaios foi estudado o crescimento bacteriano, o comportamento do pH e a degradação de cianeto. A quantificação do cianeto livre foi determinada pelo método polarográfico com eletrodo de mercúrio. Através dos resultados obtidos, foi possível identificar que as três cepas isoladas pertencem ás espécies Bacillus pumilus, Bacillus licheniformis e Bacillus subtilis. As cepas bacterianas apresentaram maior produção celular quando cultivadas em meio L.B a pH 7, velocidade de rotação de 190rpm e 37oC de temperatura. A máxima faixa de adaptação a ambientes alcalinos aconteceu em valores de pH 10. As melhores taxas de degradação de cianeto para a cepa B. subtilis e B. pumilus ocorreram em temperatura de 27oC e 65rpm, conseguindo degradar 100% do cianeto. A cepa B. licheniformis apresentou a melhor taxa de degradação de cianeto em temperatura de 32oC e 190rpm obtendo 99,5% de degradação. Através dos resultados obtidos no presente trabalho, foi possível avaliar o potencial de degradação de cianeto para as bactérias B. pumilus, B. licheniformis e B. subtilis as quais podem ser utilizadas como alternativa de tratamento em efluentes contaminados com cianeto. / Cyanide is a highly toxic compound that can be naturally found in the environment or as a result of anthropogenic activities such as gold mining and electroplating industry. Among the different species of the compound, hydrogen cyanide (HCN) is considered the most toxic, even concentrations of 100 ppm are lethal to humans. Degradation of this cyanide compound to less toxic compounds can be carried out through different methods such as chemical, physical and biological treatments. The present work investigated the bacterial degradation capacity of cyanide by isolated native strains from gold mining tailings. Three strains were selected from a group of twenty previously isolated strains which were identified using MALDI-TOF and 16s gene sequencing techniques. The bacterial activation and cellular growth were performed to determine the growth parameters for the strains, such as pH, temperature, rotation speed and culture medium. After the activation, the strains were adapted to grow in alkaline environments. During this phase, cellular growth was carried out in agitated flask at different pHs (7, 8, 9, 10 and 11). The adapted strains were used to perform cyanide degradation tests at pH 10 in agitated flask by using 100mL of a synthetic solution. The solution was composed of 500 ppm of potassium cyanide and 0,2mL of bacterial inoculum. During the experiments, three temperatures were evaluated (37, 32 and 27°C). Bacterial growth, cyanide degradation and pH behaviour were studied Free cyanide quantification was determined by polarographic method with a mercury electrode. It was found that the three isolated strains belong to Bacillus pumilus, Bacillus licheniformis and Bacillus subtilis groups. The highest bacterial growth was observed when the strains were cultivated in a L.B media at pH 7,0, 37°C by using 190 rpm of rotation velocity. The maximum range of adaptation to alkaline environments occurred in pH values of 10. The best cyanide degradation rate (100%) were achieved at 27°C for the strain B. suptilis and B. pumilus. The strain. B. licheniformis showed the best cyanide degradation rate (99,5%) at 37°C. The results obtained in the present work were able to evaluate the potential of cyanide degradation for the bacteria B. pumilus, B. licheniformis and B. subtilis. These results can be used as an alternative to treat wastewaters that are polluted with cyanide.

Endo-b-1,4 xylanases (EC. 3.2.1.8) are typically produced as a mixture of different hydrolytic enzymes such as b-1,4-xylosidase (EC. 3.2.1.37) , a-Larabinofuranosidases (EC. 3.2.1.55), and feruloyl esterase (EC 3.1.1.73) that hydrolyze xylan molecule, which constitutes 20-30% of the weight of wood and agricultural wastes. Thus, xylan, a renewable biomass, can be utilized as a substrate for the preparation of many products such as fuels, solvents and pharmaceuticals. Besides, xylanolytic enzymes themselves are also used in food,feed, textile industries and pre-bleaching of kraft. In the first part of the study, xylanolytic systems of a soil isolate Bacillus pumilus SB-M13 and a thermophilic fungus Scytalidium thermophilum were investigated. Production rate and type of xylanolytic changed depending on the carbon source and the microorganism. However, xylanolytic enzyme production was found to be sequential, in synergy and under the control of carbon catabolite repression for both microorganisms. In the second part, B. pumilus SB-M13 b-1,4 xylanase was purified and biochemically characterized. The enzyme was stable at alkaline pHs and highest activity was observed at 60&deg / C and pH 7.5. Enzyme Km and kcat values were determined as 1.87 mg/ml and 43,000 U/mg, respectively. B. pumilus SB-M13 and S .thermophilum a-L-arabinofuranosidases were also purified and biochemically characterized. Although produced from a mesophilic microorganism, B. pumilus SB-M13 arabinofuranosidase was quite thermostable. Moreover, unlike other fungi, S. thermophilum produced alkaline stable arabinofuranosidases. Both enzymes were multimeric, alkaline stable and most active at 70&deg / C and pH 7.0. However, when compared to S. thermophilum, catalytic power of B. pumilus SB-M13 arabinofuranosidase was higher.