Orientador: Wagner Bettiol / Banca: Antonio carlos Maringoni / Banca: Itamar Soares de Melo / Resumo: Experimentos sob condições controladas e testes de campo com bactérias endofíticas isoladas de folhas e caules de Coffea arabica e Coffea robusta, provenientes das cidades de Pedreira-SP, Mococa-SP e Pindorama-SP, foram realizadas com o objetivo de avaliar a inibição da germinação de urediniosporos de Hemileia vastatrix, raça II e o controle do desenvolvimento da ferrugem alaranjada do cafeeiro, em testes em discos de folhas, folhas destacadas e em plantas da cv. Mundo Novo, pela aplicação de suspensões de bectérias endofíticas 72 e 24 horas antes e após à inoculação de H. vastatrix e concomitantemente à inoculação com o patógeno. Isolados de bactérias endofíticas testados demonstraram eficácia na inibiação da germinação de urediniosporos e/ou no desenvolvimento da ferrugem, com valores acima de 50%, embora os resultados obtidos nos testes de germinação de urediniosporos tenham sido inferiores ao tratamento com o propiconazole. Nos testes em discos de folhas, folhas destacadas e em plantas de cafeeiro, os isolados endofíticos T G4-Ia, T F2-IIc, T F7-Ib e T F9-Ia, demonstraram melhor controle da ferrugem do cafeeiro. Os isolados endofíticos T G4-Ia e T F9-Ia foram identificados como Bacillus lentimorbus e Bacillus cereus, respectivamente. Os resultados indicam a ocorrência de isolados endofíticos eficientes em controlar a ferrugem alaranjada do cafeeiro. Alguns isolados de bactérias edonfíticas podem aumentar a severidade da doença. / Abstract: Experiments under controlled conditions and field tests with endophytic bacteria from leaves and stem of Coffea arabica and Coffea robusta plants from Pedreira-SP, Mococa-SP and Pindorama-SP, were carried out with th objective to evaluate the inhibition of germination of urediniospores of Hemileia vastatrix, race II and control the development of coffea leaf rust cv. Mundo Novo leaf disk, datached-leaf and coffee plants, by dripping of bacterial suspensions 72 e 24 hours prior and after the H. Vastatrix and also concomitant the aplication of the pathogen. Endophytes tested demonstrated inhibition efficacy of urediniospores germination and/or the development of leaf rust over 50%, althought the results obtained in urediniospores germination was below of propiconazole treatment. In leaf dsk and datached-leaf inoculations, the endophytes strains T G4-Ia, T F2-IIc, T F7-Ib and T F9-Ia demonstrated better control of coffee leaf rust disease. The endophytic strains T G4-Ia and T F9-Ia was identified as Bacillus lentimorbus and Bacillus cereus. The results the ocorrence of efficiente endophytes controlling the coffee leaf rust. Some endophytes can partially improve severity of coffee leaf rust. / Mestre

Orientador: Carlos Alexandre Costa Crusciol / Banca: Gabriela Ferraz de Siqueira / Banca: Rafaela Rossetto / Banca: Gustavo Pavan Mateus / Banca: Sergio Gustavo Quassi de Castro / Resumo: Inúmeros são os patógenos que reduzem a produção da cana-de-açúcar, em particular os fitonematoides apresentam grande importância. Em campo, os sintomas de ataque de nematoides são reboleiras de plantas raquíticas e cloróticas, murchas nas horas mais quentes do dia e menos produtivas. Na tentativa de diminuir as populações de nematoides abaixo do nível de dano econômico, vários métodos de controle, em integração, têm sido pesquisados, dentre eles está o controle químico, biológico e adição de matéria orgânica ao solo. Objetivou-se, por meio do presente estudo, avaliar os efeitos estimulantes de crescimento de plantas de cana-de-açúcar pela aplicação de Trichoderma asperellum, Bacillus subtilis e Bacillus methylotrophicus no controle biológico de nematoides, em área sem e com aplicação de torta de filtro. O experimento foi conduzido em cana planta, soca e soca de 4o corte, nos anos agrícola de 2015/16 e 2016/17, em áreas pertencente à Usina da Barra, Grupo Raízen Energia S/A. O delineamento experimental utilizado foi em blocos casualizados, com quatro repetições: sendo em esquema fatorial 2x5, para cana planta e soca, constituído do fator torta de filtro (1 - sem e 2 - com) combinado com o fator nematicida (1 - controle, 2 - carbofurano, 3 - Quality + Rizos, 4 - Quality + Onix, 5 - Quality + Onix + Rizos), e apenas do fator nematicida para cana soca de 4o corte. As avaliações realizadas foram: teores foliares de macronutrientes, avaliações biométricas (altura, diâmetro, número... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: There are many pathogens that reduce the sugarcane production, especially the phytonematodes are of great importance. In the field, the nematode attack symptoms are the formation of reeds and stunted and chlorotic plants, wilted in the hottest hours of the day and less productive. In order to reduce nematode populations to a minimum level of economic damage, several integration methods have been researched, including chemical, biological control and addition of organic matter to the soil. The objective of this study was to evaluate the growth stimulating effects of sugarcane plants by the application of Trichoderma asperellum, Bacillus subtilis and Bacilus methylotrophicus in the biological control of nematodes, in an area without and with application of filter cake. The experiment was carried out in sugarcane plant, sugarcane ratoon and 4th ratoon, in the agricultural years of 2015/16 and 2016/17, in areas belonging to the Usina da Barra, Grupo Raízen Energia S/A. The experimental design was a randomized complete block design, with four replications: a 2x5 factorial scheme for sugarcane plant and ratoon, consisting of filter cake factor (1 - without and 2 - with) combined with nematicidal factor (1 - control, 2 - carbofuran, 3 - Quality + Rizos, 4 - Quality + Onix, 5 - Quality + Onix + Rizos), and only the nematicidal factor for sugarcane 4th ratoon. The evaluations were: foliar macronutrient contents, biometric evaluations (height, diameter, number of stalks per meter, mean internode length, number of internodes and shoot yield) and technological (pol, purity, fiber, reducing sugar, total sugar recoverable and sugar production). The incorporation of filter cake into the soil contributed to increases in crop productivity through the addition ... / Doutor

A total of twenty-eight Bacillus spp. isolated from value-added surimi nuggets and their raw ingredients, were tested against each other and selected reference strains of Bacillus and Clostridium for their production of inhibitory substances using the deferred antagonism assay plating method. The isolated Bacillus strains showed inhibitory activity against all Bacillus strains, with the exception of the producer strain, as well as being effective against various strains of C. botulinum (type A, B and E). Subsequent studies showed that the inhibitory activity was detected in the culture supernatant in the late stationary phase of growth prior to sporulation. The inhibitory activity of two Bacillus strains (FN2A and FN33) were selected for further study. The inhibitory substances produced by these two strains were proteinaceous in nature, heat stable (100°C for 15min) and unaffected by organic solvents. A comprehensive study was conducted on the structural characterization of the inhibitor produced by B. subtilis FN2A using FPLC, FTIR, MS and MS/MS. Structural analysis of the inhibitor produced by B. subtilis FN2A showed that it was similar in structure to Surfactin. / Preliminary studies have shown that the Surfactin-like-compound from B. subtilis FN2A was produced in significant amounts during growth in bread with maximum production occurring in the late stationary phase (72h), at 30--35°C and at pH 6.5--7.0. Optimization studies on the production of the Surfactin-like-compound by B. subtilis FN2A in bread using a response surface methodology approach showed that temperature (33--36°C); autoclaving time (30 min); inoculum level (4%), alkali pre-treatment (0.16%), water activity (0.995) and pH 6.66 enhanced the production of the Surfactin-like-compound in bread. The compound produced under these optimal conditions also maintained its activity when subjected to various processing treatments (autoclaving, freezing and freeze drying). / Initial studies showed that low levels (1% w/w) of the Surfactin-like-compound inhibited the growth of B. cereus and proteolytic and non-proteolytic strains of C. botulinum in a model agar system. However, it had no effect on non-proteolytic strains of C. botulinum when bread, or methanol extracts of bread (1--20%), were added to formulated value-added sterile trout nuggets, with all nuggets being toxic after 28 days at 12°C. Furthermore, inoculation of B. subtilis FN2A directly into nuggets also failed to inhibit growth of non-proteolytic strains of C. botulinum. Omitting certain ingredients in the formulation failed to enhance the anti-botulinal effect of the bread or methanol extracts of the Surfactin-like-compound in the value-added nuggets. However, reducing the pH of the nuggets to ~5.5 enhanced the anti-botulinal effect of the Surfactin-like-compound. Further research is required to improve the dispersibility of the Surfactin-like-compound to inhibit the growth of C. botulinum in food systems.

High pressure (HP) sterilization of low acid foods is a process involving HP at elevated temperatures. Data available on HP destruction kinetics of pressure resistant pathogenic, spoilage-causing and surrogate bacterial spores are limited, but essential for establishing such a process. While compression heating could be employed to provide the elevated temperature, accurate data gathering under such conditions is difficult due to rapid heat loss during the treatment. The objectives of this research were to first standardize the set up carrying out HP tests at elevated and stabilized temperatures, and subsequently to evaluate HP destruction kinetics of selected bacterial spores under HP elevated temperature processing conditions. / The set-up consisted of a thick insulated chamber for holding samples during the test. A relationship was established for the adiabatic temperature rise milk which was used as the main low acid food medium: DeltaTP = -0.306 + 0.0224Ti + 0.0423P + 4.49x10-4T i2 + 1.31x10-4TiP -- 1.24x10 -5P2 (R2 =0.999, n = 50, SE = 0.20°C, p<0.05). Initial temperatures at 83.5, 91.7, 95.8°C and 80.7, 88.9, 93.0°C provided operating process temperature 121, 130, 135°C, at 800, 900MPa, respectively. / Destruction kinetics tests were carried with two strains of Clostridium sporogenes (11437, 7955) and Geobacillus stearothermophilus 10149 spores suspended in milk at 700-900 MPa and 70-100°C. These strains were selected for their relatively high pressue and thermal resistance. The survival counts were well fitted by first order linear models. The D values C. sporogenes 11437 varied from 0.73 min at 900 MPa 100°C to 17.0 min at 700 MPa 80°C HP treatments while they ranged from 6.0 to 833 min at 80-100°C under thermal processing conditions. The D values associated with of C. sporogenes 7955 spores were higher and varied from 1.3 min at 900 MPa 100°C to 38.2 min at 700 MPa 80°C HP treatments, and from 12.1 to 156 min at 80-100°C during thermal treatments. The D values of Geobacillus stearothermophilus 10149 spores varied from 0.6 min at 900 MPa 90C to 20.9 min at 500 MPa 70°C HP treatments with 6.3 to 49.4 min for thermal treatments at 110-120°C. Hence C. sporogenes 7955 spores were the most resistant among those studied. The HP destruction kinetics of C. sporogenes 7955 spores were also studied in salmon and were lower than in milk. / Pressure resistance screening of selected group I C. botulinum spores was carried out at 800-900 MPa at 90-100°C. Nominal D values of each strain were evaluated which demonstrated that PA9508B, H09504A and CK2-A were the more resistant spores (PA9508B > HO9504A > CK2-A). HP destruction kinetics of C. botulinum PA9508B spores in milk were evaluated in detail at 700-900 MPa at 90-100°C and parallel thermal treatment at 90-100°C. The survival counts were again described by first order linear models (R2 >0.86). The D values varied from 0.35 min at 900 MPa 110°C to 38.9 min at 700 MPa 90°C for HP treatments and 14.4 to 273 min at 90-100°C for thermal treatments. These demonstrated that HP processing combined with elevated temperatures will accelerate the spore destruction rate. However, the associated D values of C. botulinum PA9508B spores in milk were more resistant than those of C. sporogenes 7955 at 90, 100°C, which indicated that the surrogate may not be very effective for using as a target for verification HP at elevated temperature processing conditions. / Overall, this work has demonstrated several findings. The non-pathogenic C. sporogenes 7955 spore was the most resistant surrogate but the pathogenic C. botulinum PA9508B spore was even more resistant. D values associated with HP at elevated temperatures were higher than under conventional thermal treatments, and hence provide accelerated destruction kinetics at least for the non-pathogenic spores and hence better spoilage control. However, from safety point of view the conventional thermal sterility requirements would still persist even under HP processing conditions. Milk as a low acid food medium provided more resistance for HP destruction than fish.

Made available in DSpace on 2014-06-11T19:28:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2004-04-15Bitstream added on 2014-06-13T19:16:45Z : No. of bitstreams: 1 shiomi_hf_me_botfca.pdf: 1002815 bytes, checksum: aa3a55c9b04258872a970f0e010f3cdf (MD5) / Universidade Estadual Paulista (UNESP) / Experimentos sob condições controladas e testes de campo com bactérias endofíticas isoladas de folhas e caules de Coffea arabica e Coffea robusta, provenientes das cidades de Pedreira-SP, Mococa-SP e Pindorama-SP, foram realizadas com o objetivo de avaliar a inibição da germinação de urediniosporos de Hemileia vastatrix, raça II e o controle do desenvolvimento da ferrugem alaranjada do cafeeiro, em testes em discos de folhas, folhas destacadas e em plantas da cv. Mundo Novo, pela aplicação de suspensões de bectérias endofíticas 72 e 24 horas antes e após à inoculação de H. vastatrix e concomitantemente à inoculação com o patógeno. Isolados de bactérias endofíticas testados demonstraram eficácia na inibiação da germinação de urediniosporos e/ou no desenvolvimento da ferrugem, com valores acima de 50%, embora os resultados obtidos nos testes de germinação de urediniosporos tenham sido inferiores ao tratamento com o propiconazole. Nos testes em discos de folhas, folhas destacadas e em plantas de cafeeiro, os isolados endofíticos T G4-Ia, T F2-IIc, T F7-Ib e T F9-Ia, demonstraram melhor controle da ferrugem do cafeeiro. Os isolados endofíticos T G4-Ia e T F9-Ia foram identificados como Bacillus lentimorbus e Bacillus cereus, respectivamente. Os resultados indicam a ocorrência de isolados endofíticos eficientes em controlar a ferrugem alaranjada do cafeeiro. Alguns isolados de bactérias edonfíticas podem aumentar a severidade da doença. / Experiments under controlled conditions and field tests with endophytic bacteria from leaves and stem of Coffea arabica and Coffea robusta plants from Pedreira-SP, Mococa-SP and Pindorama-SP, were carried out with th objective to evaluate the inhibition of germination of urediniospores of Hemileia vastatrix, race II and control the development of coffea leaf rust cv. Mundo Novo leaf disk, datached-leaf and coffee plants, by dripping of bacterial suspensions 72 e 24 hours prior and after the H. Vastatrix and also concomitant the aplication of the pathogen. Endophytes tested demonstrated inhibition efficacy of urediniospores germination and/or the development of leaf rust over 50%, althought the results obtained in urediniospores germination was below of propiconazole treatment. In leaf dsk and datached-leaf inoculations, the endophytes strains T G4-Ia, T F2-IIc, T F7-Ib and T F9-Ia demonstrated better control of coffee leaf rust disease. The endophytic strains T G4-Ia and T F9-Ia was identified as Bacillus lentimorbus and Bacillus cereus. The results the ocorrence of efficiente endophytes controlling the coffee leaf rust. Some endophytes can partially improve severity of coffee leaf rust.

Made available in DSpace on 2014-06-11T19:32:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-07-12Bitstream added on 2014-06-13T21:05:13Z : No. of bitstreams: 1 blanco_kc_dr_rcla_parcial.pdf: 165788 bytes, checksum: 441eba3e7f81a4045fd71fd169810c38 (MD5) Bitstreams deleted on 2015-06-03T11:42:26Z: blanco_kc_dr_rcla_parcial.pdf,. Added 1 bitstream(s) on 2015-06-03T11:43:59Z : No. of bitstreams: 1 000722980_20150710.pdf: 153337 bytes, checksum: ef6f673ca2880afc3824d702fd97308c (MD5) Bitstreams deleted on 2015-08-03T12:21:09Z: 000722980_20150710.pdf,. Added 1 bitstream(s) on 2015-08-03T12:22:22Z : No. of bitstreams: 1 000722980.pdf: 702124 bytes, checksum: 076acfb344282eec1bfa2c15d2efa0a1 (MD5) / As ciclodextrinas (CDs) resultam da degradação do amido por ciclodextrina glicosiltransferase (CGTases) produzidas principalmente pelo gênero Bacillus. As CDs são oligossacarídeos cíclicos capazes de formar complexos de inclusão com uma grande variedade de moléculas modificando suas características indesejadas sendo assim muito utilizadas em vários setores industriais. A linhagem bacteriana Gram-positiva e formadora de esporos nomeada como CGII foi isolada de águas residuárias de uma fábrica de farinha de mandioca em Ribeirão Bonito, São Paulo, Brasil e submetidos a estudos filogenéticos e testes bioquímicos. O planejamento composto central foi então utilizado para optimizar a composição do meio e as condições de cultivo para a produção da enzima ciclodextrina glicosiltransferase (CGTase) em frascos agitados e em biorreatores. As características físico-químicas e bioquímicas da CGTase como pH e temperatura óptima; estabilidade em temperatura e pH; a influência de substâncias; cinética enzimática e produção de ciclodextrina foram avaliadas após purificação em cromatografia de afinidade usando a β-CD como ligante. Neste trabalho também foram investigadas novas aplicações da ciclodextrina produzida pela CGTase de B. lehensis: A síntese de fases sólidas de nanopartículas de óxido de ferro magnético (Fe4O3), óxido de cobre (CuO) e prata metálica (Ago) a partir de CDs por síntese hidrotérmica, usando CDs. As nanopartículas foram caracterizadas por difração de raios-x, espectroscopia de infravermelho e microscopia eletrônica de varredura. A enzima CGTase também foi imobilizada por ligação covalente no suporte magnetita síntetizada anteriormente, a qual foi silinalizado com 3-aminopropiltrimetilsilano e ativada com glutaraldeido. A sequência do gene 16S rRNA apresentou maior grau de similaridade com... / Cyclodextrins (CDs) are obtained from starch degradation by enzyme cyclodextrin glycosyltransferase (CGTases) produced mainly by Bacillus genus. The CDs are cyclic oligosaccharides capable of forming inclusion complexes with a wide variety of molecules, modifying their unwanted characteristics and, therefore are widely used in various industrial sectors. The bacterial strain Gram-positive and spore-forming named CGII was isolated from wastewater from a cassava mill in São Paulo, Brazil and subjected to phylogenetic studies and biochemical tests. A central composite design was then used to optimize the medium composition and culture conditions for the production of the enzyme cyclodextrin glycosyltransferase (CGTase) in bioreactors. The physicochemical and biochemical proprieties of CGTase as pH and temperature optimum, stability to pH and temperature, the influence of substances, enzyme kinetics and production of cyclodextrin were evaluated after purification by affinity chromatography using β-CD as a ligand. In this work new applications of cyclodextrin produced by CGTase from B. lehensis were also investigated. The solid phase synthesis of nanoparticles magnetic iron oxide (Fe4O3), copper oxide (CuO) and metallic silver (Ag) by hydrothermal synthesis using CDs were characterized by x-ray, infrared, and scanning electron microscopy. The enzyme CGTase was also immobilized on the support by covalent bond to the synthesized magnetite, which has been silylazed with 3- aminopropil-trimetilsilano and activated with glutaraldehyde. The 16S rRNA gene sequence indicated the highest degree of similarity to strains of Bacillus genomic MLB2 lehensis (100%). The response surface methodology showed that the model for the optimization of CGTase of B. lehensis reached a good level of agreement with experimental... (Complete abstract click electronic access below)

Orientador: Ana Maria Centola Vidal / Banca: Karina Paes Bürger / Banca: Andréia Cristina Nakashima Vaz / Resumo: O tratamento térmico por UAT elimina formas vegetativas dos microrganismos, porém, formas esporuladas altamente resistentes ao calor podem permanecer nos produtos e após a germinação dos esporos são capazes de liberar toxinas causadoras de doenças transmitidas por alimentos (DTAs). No presente estudo, objetivou-se avaliar a qualidade microbiológica de produtos tratados por UAT, por meio da contagem de microrganismos heterotróficos aeróbios mesófilos, isolamento e contagem de bactérias do grupo do Bacillus cereus e de Clostridium perfringens, comparar lotes dos três produtos quanto a tais microrganismos, identificar a espécie dos isolados e a presença dos fatores de virulência de bactérias do grupo do B. cereus. Para isso, foram obtidas 75 amostras, sendo 25 de cada produto (de leite de cabra UAT, alimento com soja UAT e bebida de leite de cabra e soja UAT), de 5 diferentes lotes, no comércio do interior do Estado de São Paulo. Os resultados evidenciaram possíveis falhas de processamento e utilização de matéria-prima inadequada, uma vez que altas populações (>104UFC/mL) de microrganismos heterotróficos aeróbios mesófilos foram verificadas em 100% dos lotes de alimento com soja e bebida de leite de cabra e soja e 80% dos lotes de leite de cabra. Microrganismos do grupo do B. cereus foram isolados de 80% dos lotes de alimento com soja, e 60% dos lotes de bebida de leite de cabra e soja e leite de cabra, com populações superiores a 1018 UFC/mL. C. perfringens foi isolado de 20% d... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Heat treatment by UHT (ultra-high temperature) eliminates the vegetative forms of microorganisms, however, sporulated forms highly resistant to heat can remain in the products and following spore germination are capable of releasing toxins that cause foodborne diseases (FD's). The objective of this study is to evaluate the microbiological quality of products submitted to UHT, namely goat milk, soy food, and goat and soy milk drinks, by the counting of heterotrophic aerobic mesophilic microorganisms, the isolation and counting of bacteria from the Bacillus cereus groups and Clostridium perfringens, comparing batches of the three products for such microorganisms, to identify the species of the isolated and the presence of the virulence factors of bacteria of the B. cereus group. For this, 75 samples were obtained, 25 of each product (from UAT goat milk, food with UAT soy and drink from goat milk and UAT soy) from 5 different lots, in the interior trade of the State of São Paulo. The results demonstrated possible failures in the processing and use of inadequate raw material since high population (>104 ) of heterotrophic aerobic mesophilic microorganisms were found in 100% of the lots of soy food and goat and soy milk drinks and 80% of the lots of goat milk. Microorganisms from the B. cereus group were isolated from 80% of the lots of soy food and 60% of the lots of goat and soy milk drinks, with population above 1018 . The C. perfringens microorganisms were isolated from 20% of the soy food lots, with population above 103 . In the comparison between the lots, the expiration date did not influence the population of the analyzed microorganisms, therefore, they were conveyed by the raw material used. Of the B. cereus isolated, 100% (29/29) showed amplification for the hbl complex gene, 96,5% (28/29) for the nhe complex gene and 75,8% (22/29) for t... Complete abstract click electronic access below / Mestre

Made available in DSpace on 2015-06-17T19:34:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-26. Added 1 bitstream(s) on 2015-06-18T12:49:23Z : No. of bitstreams: 1 000829428_20170301.pdf: 100482 bytes, checksum: 894af24aad4ca7d75f9898581c07f398 (MD5) Bitstreams deleted on 2017-03-03T11:01:31Z: 000829428_20170301.pdf,. Added 1 bitstream(s) on 2017-03-03T11:02:40Z : No. of bitstreams: 1 000829428.pdf: 275587 bytes, checksum: 79098a3390015f4449a4b64413d932a8 (MD5) / Estudos que visam a interação das proteínas Cry de Bacillus thuringiensis, a fim de encontrar combinações adequadas para o desenvolvimento de plantas Bt são ferramentais fundamentais no controle de lepidópteros-praga. A lagarta H. armigera causa danos severos nas culturas agrícolas e sua introdução no Brasil levou a busca de formas de controle eficientes e nesse contexto B. thuringiensis pode ser um bom agente de controle. Diante do exposto o presente trabalho objetivou avaliar a toxicidade das proteínas Cry1Aa, Cry1Ab, Cry1Ac e Cry1Ca de B. thuringiensis à H. armigera, assim como interação dessas proteínas aos receptores do mesêntero do inseto. A toxicidade foi estimada com bioensaios de dose resposta com as proteínas testadas e a interação das proteínas com os receptores foram verificadas em análise de união entre a proteína ativada e marcada com a vesícula da borda em escova da membrana apical das células do intestino (brush border mambrane vesicle- BBMV) do mesêntero larval de H. armigera, e ensaios de competição heteróloga. Dentre as proteínas testadas, a Cry1Ac destacou-se como a mais efetiva, seguida das proteínas Cry1Ab e Cry1Aa. A proteína Cry1Ca não apresentou toxicidade. As proteínas Cry1Aa, Cry1Ab e Cry1Ac se ligaram aos receptores da membrana do intestino médio das lagartas de H. armigera de forma especifica. Os ensaios de competição heteróloga revelaram que as proteínas Cry1Aa, Cry1Ac e Cry1Ab competem entre si pelo mesmo receptor / Studies attempting interaction of Bacillus thuringiensis Cry proteins in order to find combinations for developing Bt plants are fundamental in controlling lepidopteran pests. H. armigera causes severe damage to agricultural crops and their introduction in Brazil has led the search for efficient control and B. thuringiensis may be a good control agent. The aim of this research was to evaluate the toxicity of Cry1Aa, Cry1Ab, Cry1Ac and Cry1Ca proteins from B. thuringiensis to H. armigera, as well as interaction of these proteins with the receptors present in insect midgut. Toxicity was estimated from the lethal concentration LC50 of the tested proteins and protein interactions with the receptors were found in a binding analysis between activated and biotinylated protein with the midgut brush border vesicle membrane (BBMV) of H. armigera, and heterologous competitive binding assays. Among the tested proteins, Cry1Ac protein was the most toxic, followed by the Cry1Ab and Cry1Aa proteins. The Cry1Ca protein showed no toxicity. The Cry1Aa, Cry1Ab and Cry1Ac proteins showed specific binding to the midgut membrane receptors of H. armigera caterpillars. Heterologous competitive binding assays revealed that Cry1Aa, Cry1Ab, Cry1Ac compete for a common receptor in the midgut larvae

Orientador: Ana Julia Fernandes / Resumo: Com o processo de expansão industrial, as regiões costeiras e estuarinas passaram a perceber a instalação de várias indústrias, devido à proximidade aos mercados consumidores e produtores, além da facilidade de escoamento de sua produção. Essas indústrias consomem uma grande quantidade de água, que é devolvida ao ambiente juntamente com uma série de substâncias tóxicas dissolvidas, incluindo hidrocarbonetos aromáticos, de biodegradabilidade intermediária e tendência à bioacumulação. Regiões estuarinas apresentam maior dificuldade de biorremediação, devido ao fato dos contaminantes ficarem aderidos ao sedimento. Além do descarte costeiro habitual, acidentes tendem a lançar uma quantidade maior e descontrolada de poluentes. A biorremediação é o processo de tratamento que utiliza os microrganismos encontrados tanto no ambiente terrestre como aquáticos contaminados para degradar substâncias toxicamente perigosas transformando-as em substâncias menos ou não tóxicas. As atividades para conter o incêndio no Terminal Químico Aratu S.A. (Ultracargo) lançou grande quantidade de hidrocarbonetos e líquido gerador de filme aquoso (AFFF) no estuário de Santos, modificando o perfil de contaminantes da região. As bactérias heterotróficas de ambientes marinhos apresentam grande potencial de biorremediação. O objetivo do presente trabalho foi avaliar, in vitro, a sucessão de espécies de bactérias heterotróficas em sedimentos do Estuário de Santos, afetados por hidrocarbonetos e AFFF e isolar e... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: With the industrial expansion, coastal and estuarine regions began to perceive installation of several industries, due to the proximity to the consumer markets and producers, besides the ease of transportation of their production. These industries consume a large amount of water, which is returned to the environment with a series of dissolved toxic substances, which includes aromatic hydrocarbons, which are slightly biodegradable and have the tendency to bioaccumulate. Estuarine regions present higher difficulty of bioremediation, since the contaminants are adhered to the sediment. In addition to the usual coastal discard, accidents tend to release a larger and uncontrolled amount of pollutants. Bioremediation is the treatment process that uses microorganisms found in both the terrestrial and aquatic environments to degrade toxic substances by transforming them into less toxic or non-toxic substances. The fire at the Terminal Químico Aratu S.A. (Ultracargo) launched a large amount of hydrocarbons and aqueous film-forming foam (AFFF) in the Santos estuary, modifying the profile of contaminants in the region. The heterotrophic bacteria of marine environments present great bioremediation potential. The objective of the present work was to evaluate in vitro the succession of heterotrophic bacteria species in Santos Estuary sediments affected by hydrocarbons and AFFF and to isolate bacterial species with potential use in bioremediation processes. The densities of heterotrophic bacteria were analyzed and the most relevant ones were identified, by morphological characteristics and molecular identification, where the Bacillus genus deserved attention. The succession suggested three species as potential bioremediators, being Bacillus pumilus, Bacillus subtilis and Staphylococcus sp... (Complete abstract electronic access below) / Mestre

Orientador: Manoel Victor Franco Lemos / Coorientador: Ricardo Antonio Polanczyk / Banca: Janete Apparecida Desidério / Banca: Camila Chiaradia Davolos / Resumo: Estudos que visam a interação das proteínas Cry de Bacillus thuringiensis, a fim de encontrar combinações adequadas para o desenvolvimento de plantas Bt são ferramentais fundamentais no controle de lepidópteros-praga. A lagarta H. armigera causa danos severos nas culturas agrícolas e sua introdução no Brasil levou a busca de formas de controle eficientes e nesse contexto B. thuringiensis pode ser um bom agente de controle. Diante do exposto o presente trabalho objetivou avaliar a toxicidade das proteínas Cry1Aa, Cry1Ab, Cry1Ac e Cry1Ca de B. thuringiensis à H. armigera, assim como interação dessas proteínas aos receptores do mesêntero do inseto. A toxicidade foi estimada com bioensaios de dose resposta com as proteínas testadas e a interação das proteínas com os receptores foram verificadas em análise de união entre a proteína ativada e marcada com a vesícula da "borda em escova" da membrana apical das células do intestino ("brush border mambrane vesicle"- BBMV) do mesêntero larval de H. armigera, e ensaios de competição heteróloga. Dentre as proteínas testadas, a Cry1Ac destacou-se como a mais efetiva, seguida das proteínas Cry1Ab e Cry1Aa. A proteína Cry1Ca não apresentou toxicidade. As proteínas Cry1Aa, Cry1Ab e Cry1Ac se ligaram aos receptores da membrana do intestino médio das lagartas de H. armigera de forma especifica. Os ensaios de competição heteróloga revelaram que as proteínas Cry1Aa, Cry1Ac e Cry1Ab competem entre si pelo mesmo receptor / Abstract: Studies attempting interaction of Bacillus thuringiensis Cry proteins in order to find combinations for developing Bt plants are fundamental in controlling lepidopteran pests. H. armigera causes severe damage to agricultural crops and their introduction in Brazil has led the search for efficient control and B. thuringiensis may be a good control agent. The aim of this research was to evaluate the toxicity of Cry1Aa, Cry1Ab, Cry1Ac and Cry1Ca proteins from B. thuringiensis to H. armigera, as well as interaction of these proteins with the receptors present in insect midgut. Toxicity was estimated from the lethal concentration LC50 of the tested proteins and protein interactions with the receptors were found in a binding analysis between activated and biotinylated protein with the midgut brush border vesicle membrane (BBMV) of H. armigera, and heterologous competitive binding assays. Among the tested proteins, Cry1Ac protein was the most toxic, followed by the Cry1Ab and Cry1Aa proteins. The Cry1Ca protein showed no toxicity. The Cry1Aa, Cry1Ab and Cry1Ac proteins showed specific binding to the midgut membrane receptors of H. armigera caterpillars. Heterologous competitive binding assays revealed that Cry1Aa, Cry1Ab, Cry1Ac compete for a common receptor in the midgut larvae / Mestre