O copolímero biodegradável poli(3-hidroxibutirato-co-3-hidroxihexanoato) P(3HB-co-3HHx) é um polihidroxialcanoato (PHA) que apresenta várias aplicações. A bactéria Burkholderia sacchari acumula P(3HB-co-2mol%3HHx), a partir de glicose e ácido hexanoico. Com o objetivo de obter P(3HB-co-3HHx) com diferentes teores de 3HHx por B. sacchari, foram construídas linhagens recombinantes, contendo genes do operon phaPCJ de Aeromonas spp. Os recombinantes produziram P(3HB-co-3HHx), a partir de ácidos hexanoico, láurico e linoleico, com teores de 3HHx entre 1,88-18 mol%. Experimentos em biorreator com o recombinante, alimentada na fase de acúmulo por glicose 140 g/L e ácido hexanoico entre 0-45 g/L, resultaram copolímeros com composições variando de 0 a 20 mol% de 3HHx. Os copolímeros assim produzidos foram extraídos e analisados quanto às propriedades físicas. A análise de fluxos metabólicos indicou que a produção de PHA pode ser aumentada com mudanças no metabolismo central e deleção/superexpressão de genes. / The biodegradable copolymer poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) P(3HB-co-3HHx) is a polyhydroxyalkanoate (PHA) presenting various applications. The bacterium Burkholderia sacchari accumulated P(3HB-co-2mol%3HHx) from glucose and hexanoic acid. In order to obtain P(3HB-co-3HHx) with different 3HHx amounts by B. sacchari, recombinant strains containing phaPCJ operon genes from Aeromonas spp were constructed. Recombinant strains produced P(3HB-co-3HHx) from hexanoic, lauric and linoleic acids, with contents of 3HHx ranging from 1.88 to 18 mol%. Experiments with the recombinant in bioreactor, fed in the accumulation phase by glucose 140 g.l-1and hexanoic acid 0-45 g.l-1, resulted in copolymers with compositions ranging from 0 to 20 mol% of 3HHx. The copolymers produced were extracted and analyzed for physical properties. The metabolic flux analysis indicated that PHA production can be increased by modifying the central metabolism and deleting/ overexpressing genes.

O copolímero biodegradável poli(3-hidroxibutirato-co-3-hidroxihexanoato) P(3HB-co-3HHx) é um polihidroxialcanoato (PHA) que apresenta várias aplicações. A bactéria Burkholderia sacchari acumula P(3HB-co-2mol%3HHx), a partir de glicose e ácido hexanoico. Com o objetivo de obter P(3HB-co-3HHx) com diferentes teores de 3HHx por B. sacchari, foram construídas linhagens recombinantes, contendo genes do operon phaPCJ de Aeromonas spp. Os recombinantes produziram P(3HB-co-3HHx), a partir de ácidos hexanoico, láurico e linoleico, com teores de 3HHx entre 1,88-18 mol%. Experimentos em biorreator com o recombinante, alimentada na fase de acúmulo por glicose 140 g/L e ácido hexanoico entre 0-45 g/L, resultaram copolímeros com composições variando de 0 a 20 mol% de 3HHx. Os copolímeros assim produzidos foram extraídos e analisados quanto às propriedades físicas. A análise de fluxos metabólicos indicou que a produção de PHA pode ser aumentada com mudanças no metabolismo central e deleção/superexpressão de genes. / The biodegradable copolymer poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) P(3HB-co-3HHx) is a polyhydroxyalkanoate (PHA) presenting various applications. The bacterium Burkholderia sacchari accumulated P(3HB-co-2mol%3HHx) from glucose and hexanoic acid. In order to obtain P(3HB-co-3HHx) with different 3HHx amounts by B. sacchari, recombinant strains containing phaPCJ operon genes from Aeromonas spp were constructed. Recombinant strains produced P(3HB-co-3HHx) from hexanoic, lauric and linoleic acids, with contents of 3HHx ranging from 1.88 to 18 mol%. Experiments with the recombinant in bioreactor, fed in the accumulation phase by glucose 140 g.l-1and hexanoic acid 0-45 g.l-1, resulted in copolymers with compositions ranging from 0 to 20 mol% of 3HHx. The copolymers produced were extracted and analyzed for physical properties. The metabolic flux analysis indicated that PHA production can be increased by modifying the central metabolism and deleting/ overexpressing genes.

Made available in DSpace on 2016-04-28T20:17:28Z (GMT). No. of bitstreams: 1 2008 - Ronaldo Leao Guimaraes.pdf: 352046 bytes, checksum: 7639df2176f027af3722d0b90e93a856 (MD5) Previous issue date: 2008-12-01 / The aim of this study was to evaluate the presence of Vibrio sp., Aeromonas sp. and Enterobacteriaceae in marine and farmer shrimp. For this, 20 samples of marine shrimp (Penaeus sp.) were collected from Itajuru channel, Cabo Frio, RJ, and other 20 samples of farmer shrimp were acquired in municipal market of Cabo Frio, RJ between January to August 2007. These samples were processed using standard methodology and inoculated in selective media for the research of these microorganisms. The isolates identification was performed by specific biochemical tests. 88 strains were isolated; 51.1% belonging to Vibrio sp., 28.5% to Aeromonas sp. and 20.4% to Plesiomonas shigelloides. It was not detected the presence of E. coli or Salmonella. Among the Vibrio sp., there was a prevalence of Vibrio alginolyticus with 45% and Vibrio parahaemolyticus with 10.5%. Aeromonas hydrophila contributed with 30% of the isolates of this genus and 83.3% of the isolates from Plesiomonas shigelloides were obtained from marine shrimp while 16.7% were obtained from farmer shrimp. The results show a high frequency of potentially pathogenic strains in these shrimps, emphasizing the importance of monitoring these organisms not only to shrimp industry but also to public health. / Este trabalho teve como objetivo pesquisar em camar?es de vida livre e criados em cativeiro a presen?a de microrganismos dos g?neros Vibrio, Aeromonas e representantes da fam?lia Enterobacteriaceae. Para tal, foram coletadas 20 amostras de camar?o de vida livre (Penaeus sp.) do canal de Itajuru, Cabo Frio, RJ; e outras 20 amostras de camar?o de cativeiro do com?rcio local deste munic?pio, no per?odo de Janeiro a Agosto de 2007. Estas amostras foram processadas seguindo metodologia padr?o e inoculadas em meios seletivos para a pesquisa dos microrganismos citados. A identifica??o dos isolados foi realizada atrav?s de testes bioqu?micos espec?ficos. 88 cepas foram isoladas, das quais, 51,1% pertencente ao g?nero Vibrio, 28,5% ao g?nero Aeromonas e 20,4% a Plesiomonas shigelloides. Em nenhuma amostra foi detectada a presen?a de E. coli ou Salmonella spp. Dentre os Vibrios, houve predomin?ncia de Vibrio alginolyticus com 45% e Vibrio parahaemolyticus com 10,5%. Aeromonas hydrophila contribuiu com 30% dos isolados deste g?nero e 83,3% dos isolados de Plesiomonas shigelloides foram obtidos de camar?o de vida livre enquanto 16,7% foram obtidos de camar?o de cativeiro. Os resultados apresentam uma grande freq??ncia de cepas potencialmente patog?nicas ao homem presentes nestes camar?es; ressaltando a import?ncia do monitoramento destes organismos n?o s? para a ind?stria de carcinicultura como tamb?m para a sa?de p?blica.

Aeromonas hydrophila is an opportunistic pathogen that causes a wide range of symptoms and diseases in fish. Development of a commercial vaccine has been problematic due to the heterogenicity between isolates of A. hydrophila. A new approach using immunoproteomics was used in this study to try to develop a vaccine that would protect against a wide range of A. hydrophila strains. The virulence of 14 isolates of A. hydrophila from different geographical regions was determined in common carp (Cyprinus carpio) indicating that 6 isolates were virulent, while 8 isolates were avirulent. Expression of cellular and extracellular products (ECP) of six of these isolates (4 virulent and 2 avirulent isolates) were examined following culture of the bacterium in vitro, in tryptic soy broth, and in vivo, in dialysis tubing placed within the peritoneal cavity of carp. Two types of molecular weight cut off tubes (25 and 100 kDa) were used for the implants. Whole cell (WC), outer membrane protein (OMP) and ECPs of the bacteria grown in vitro and in vivo were analysed by 1 dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (1D SDS-PAGE). Additionally, 2D SDS-PAGE was used to analyse WC preparations of A. hydrophila grown in vitro and in vivo. The production of unique proteins and up and down-regulation of protein expression were observed in all the preparations of bacteria grown in vitro and in vivo. Unique bands were seen in the 1D SDS-PAGE at 58 and 55 kDa for WC and OMP preparations, respectively, for all the isolates cultured in vivo. Bands of increased intensity were observed at 70, 55, 50 and 25 kDa with WC preparations for the virulent isolates cultured in vivo. Analysis of WC preparations by 2D SDS-PAGE indicated differences in the expression of spots between bacteria cultured in vitro and in vivo. A number of unique spots, mostly between 30 and 80 kDa with pI values ranging from 5.0-6.0 were observed in the bacteria grown in vivo. The protein profiles of different preparations (WC, OMP, ECP) of bacteria cultured in vitro and in vivo were screened by 1D Western blot using antibodies from carp artificially infected with different isolates of A. hydrophila to identify potential vaccine candidates. The WC preparations of A. hydrophila (T4 isolate) grown in vitro were also analysed by 2D Western blot. A 50 kDa protein of A. hydrophila was found to be the most immunogenic molecule in both WC and OMP of bacteria grown both in vitro and in vivo. The protection efficacy of this protein was determined in goldfish by vaccinating fish with electro-eluted 50 kDa protein then challenging the fish with A. hydrophila. Fish were also passively immunised with fish sera raised to the 50 kDa protein and then challenged. The relative percentage survival (RPS) was 67 % in the vaccination trial, while the results were inconclusive for the passive immunisation trial. The 50 kDa protein was confirmed to be the S-layer protein of A. hydrophila following identification using matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS). Recombinant S-layer protein was then produced and the cross-protection efficacy of this protein against six virulent isolates of A. hydrophila was confirmed in a large scale vaccination trial using carp. The RPS value for the 6 isolates of A. hydrophila ranged from between 56 and 87 %. The results of this project suggest that the immunogenic S-layer protein of A. hydrophila could be used as a common antigen to protect fish against infection by different isolates of this pathogenic bacterium.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2002. / Härtill 4 uppsatser.

Thesis (M.S.)--Miami University, Dept. of Chemistry and Biochemistry, 2004. / Title from first page of PDF document. Includes bibliographical references (p. 57-64).

Made available in DSpace on 2014-06-11T19:32:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-01-14Bitstream added on 2014-06-13T21:05:30Z : No. of bitstreams: 1 santos_vg_dr_botfmvz.pdf: 418050 bytes, checksum: ff8ea12e28e936c31b32dcbb33f589a4 (MD5) / Avaliou-se a ação imunoestimulante da própolis bruta em dietas para tilápia-do-Nilo submetidas a desafio com a Aeromonas hydrophila. Foram alojados 280 peixes (50 ± 5,7 g) em 35 aquários circulares (250 L), oito peixes/aquário. O delineamento estatístico utilizado foi inteiramente casualizado, caracterizado por sete tratamentos, dieta controle e as demais com suplementação de própolis de 0,5; 1,0; 1,5; 2,0; 2,5 e 3,0% com cinco repetições. Os peixes foram pesados no início e final do experimento para avaliação dos parâmetros de desempenho produtivo. Após trinta dias, 16 peixes de cada tratamento foram anestesiados com benzocaína (1,0g/15L de água) para coleta do sangue e avaliação dos parâmetros hematológicos e imunológicos, caracterizando período anterior ao desafio bacteriano. Os parâmetros hematológicos e imunológicos avaliados foram: número de eritrócitos, porcentagem de hematócrito, taxa de hemoglobina, volume corpuscular médio (VCM), concentração de hemoglobina corpuscular média (CHCM) e peróxido de hidrogênio e óxido nitrico (NO). Em seguida, oito peixes de cada tratamento foram inoculados com injeção intraperitoneal com cultura da bactéria contendo 2,0x107UFC/mL e transferidos para 28 aquários plásticos (40L), dois peixes por aquário. Os mesmos parâmetros foram avaliados após 15 dias. Os resultados demonstraram que níveis elevados de própolis bruta influenciaram na redução do ganho de peso e da hemoglobina e aumento do NO. Comparando-se os momentos, observou-se que após o desafio, os peixes apresentaram valores menores de hematócrito, VCM, neutrófilos e NO e maior CHCM. Concluiu-se que a própolis bruta interferiu no crescimento e na saúde da tilápia-do-Nilo / Immunostimulant effects of crude propolis supplementation on diets to Nile tilapia, submitted to Aeromonas hydrophila, were investigated. Thus, 280 Nile tilapia (initial weight = 50 ± 5,7 g/fish), were allocated in 35 tanks (250 l), eight fish/tank. The experiment was conducted in a completely randomized design, characterized by seven treatments, diet control and the other with propolis supplementation of 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0% with five repetitions. The fish were weighed at the beginning and final of experimental for evaluating the performance parameters. After thirty days, 16 fishes/treatment were anesthetized with benzocaine (1.0g/15L of water), and collected blood to evaluation to haematological and immunological parameters, characterizing the period before to bacterial challenge. The parameters analized were: red blood cell count , haematocrit, haemoglobin, mean corpuscular volume (MCV), mean corpuscular haemoglobin concentration (MCHC), and hydrogen peroxide and nitric oxide (NO). Then, eight fish of each treatment were inoculated with intraperitoneal injection with culture of A. Hydrophila containing 2.0x107UFC/mL, and transferred to twenty eight plastic tanks (40L), and two fishes per tank. The same haematological and immunological parameters were evaluated after fifteen days. The results showed that high levels of crude propolis influence at decrease of weight gain and Hb, an increase of NO. Comparing the moments, was observed that after challenge the fishes showed low value of Htc, MCV, neutrophils and NO, also high value of MCHC. It was concluded that crude propolis affect the growth and health of the-Nile tilapia

Orientador: Margarida Maria Barros / Coorientador: Ricardo de Oliveira Orsi / Banca: Luiz Edivaldo Pezzato / Banca: Lidia Mara Ruv Carelli Barreto / Banca: Maria José Tavares Ranzani de Paiva / Banca: Igo Gomes Guimarães / Resumo: Avaliou-se a ação imunoestimulante da própolis bruta em dietas para tilápia-do-Nilo submetidas a desafio com a Aeromonas hydrophila. Foram alojados 280 peixes (50 ± 5,7 g) em 35 aquários circulares (250 L), oito peixes/aquário. O delineamento estatístico utilizado foi inteiramente casualizado, caracterizado por sete tratamentos, dieta controle e as demais com suplementação de própolis de 0,5; 1,0; 1,5; 2,0; 2,5 e 3,0% com cinco repetições. Os peixes foram pesados no início e final do experimento para avaliação dos parâmetros de desempenho produtivo. Após trinta dias, 16 peixes de cada tratamento foram anestesiados com benzocaína (1,0g/15L de água) para coleta do sangue e avaliação dos parâmetros hematológicos e imunológicos, caracterizando período anterior ao desafio bacteriano. Os parâmetros hematológicos e imunológicos avaliados foram: número de eritrócitos, porcentagem de hematócrito, taxa de hemoglobina, volume corpuscular médio (VCM), concentração de hemoglobina corpuscular média (CHCM) e peróxido de hidrogênio e óxido nitrico (NO). Em seguida, oito peixes de cada tratamento foram inoculados com injeção intraperitoneal com cultura da bactéria contendo 2,0x107UFC/mL e transferidos para 28 aquários plásticos (40L), dois peixes por aquário. Os mesmos parâmetros foram avaliados após 15 dias. Os resultados demonstraram que níveis elevados de própolis bruta influenciaram na redução do ganho de peso e da hemoglobina e aumento do NO. Comparando-se os momentos, observou-se que após o desafio, os peixes apresentaram valores menores de hematócrito, VCM, neutrófilos e NO e maior CHCM. Concluiu-se que a própolis bruta interferiu no crescimento e na saúde da tilápia-do-Nilo / Abstract: Immunostimulant effects of crude propolis supplementation on diets to Nile tilapia, submitted to Aeromonas hydrophila, were investigated. Thus, 280 Nile tilapia (initial weight = 50 ± 5,7 g/fish), were allocated in 35 tanks (250 l), eight fish/tank. The experiment was conducted in a completely randomized design, characterized by seven treatments, diet control and the other with propolis supplementation of 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0% with five repetitions. The fish were weighed at the beginning and final of experimental for evaluating the performance parameters. After thirty days, 16 fishes/treatment were anesthetized with benzocaine (1.0g/15L of water), and collected blood to evaluation to haematological and immunological parameters, characterizing the period before to bacterial challenge. The parameters analized were: red blood cell count , haematocrit, haemoglobin, mean corpuscular volume (MCV), mean corpuscular haemoglobin concentration (MCHC), and hydrogen peroxide and nitric oxide (NO). Then, eight fish of each treatment were inoculated with intraperitoneal injection with culture of A. Hydrophila containing 2.0x107UFC/mL, and transferred to twenty eight plastic tanks (40L), and two fishes per tank. The same haematological and immunological parameters were evaluated after fifteen days. The results showed that high levels of crude propolis influence at decrease of weight gain and Hb, an increase of NO. Comparing the moments, was observed that after challenge the fishes showed low value of Htc, MCV, neutrophils and NO, also high value of MCHC. It was concluded that crude propolis affect the growth and health of the-Nile tilapia / Doutor

Microcystin-LR is a hepatotoxin produced by cyanobacteria. Aeromonas hydrophila and Edwardsiella piscicida infections are leading causes of losses in market-sized channel catfish (Ictalurus punctatus). These older fish should have natural immunity in place and a predisposing factor is likely a prerequisite for these disease outbreaks. While microcystin-LR rarely causes mortality in warm-water aquaculture, we believe it may be a predisposing factor that leads to bacterial disease outbreaks during the summer months due to its ability to damage the liver. Our study investigated microcystin-LR’s effects on channel catfish susceptibility to these pathogens. We found that a sublethal dose of microcystin-LR induced substantial damage to multiple immune organs. In our challenges with both the toxin and bacteria, we saw a significant increase in mortality of fish. Our findings suggest that microcystin-LR increases channel catfish susceptibility to Aeromonas hydrophila and Edwardsiella piscicida infections.

No description available.