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Made available in DSpace on 2014-06-11T19:28:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-26Bitstream added on 2014-06-13T19:58:12Z : No. of bitstreams: 1 silva_ac_me_botfca.pdf: 1497139 bytes, checksum: 1ec5c66837f996b2bf5ed59ec09974f8 (MD5) / Fundação de Apoio à Pesquisa Agrícola (FUNDAG) / Dentre os insetos pragas mais freqüentes e limitantes à produção de frutas, incluem-se a mosca-do-mediterrâneo, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), e o gorgulho-da-goiaba, Conotrachelus psidii (Marshal) (Coleoptera: Curculionidae). As larvas desses insetos se alimentam da polpa das frutas, sendo que no final dessa fase, os insetos descem para o solo onde se enterram e atravessam a fase de pupa. O objetivo do trabalho foi avaliar a virulência de 7 nematóides dos gêneros Steinernema e Heterorhabditis contra a mosca-do-mediterrâneo, e o potencial de Heterorhabditis indica Poinar, Karunakar & David 1992 no controle desse inseto e do gorgulho-da-goiaba. Foram realizados cinco experimentos, sendo um experimento em laboratório, três em casa de vegetação e um em campo. De acordo com os resultados, a mosca-do-mediterrâneo é suscetível aos nematóides quando exposta no estágio de pré-pupa e fase de pupa com 1 dia de desenvolvimento; O nematóide H. indica IBCB n5 mostrou-se como o mais virulento contra C. capitata. No teste de campo procurando avaliar H. indica IBCB n5 contra C. capitata e contra C. psidii, todos os tratamentos diferenciaram significativamente da testemunha. O nematóide apresentou-se bem mais eficiente contra a mosca-do¬mediterrâneo, com mortalidade do inseto de 66% e 93% para as dosagens de 1 e 10 JI/cm², respectivamente, do que contra o gorgulho-da-goiaba, com mortalidade de 33% e 50%, respectivamente, tendo todos esses tratamentos diferidos significativamente da testemunha / The fruit fly, Ceratitis capitata, and the guava weevil, Conotrachelus psidii, are among the most frequent and injurious pest of fruit crops. The insects larva feed on the fruit pulp and, at the end of this phase, the insects reach the soil where they spent their pupa phase. This research aimed to evaluate the virulence of 7 strains of entomopathogenic nematodes against the fruit fly in laboratory and green house conditions, and the efficiency of Heterorhabditis indica IBCB n5 against this insect and the guava weevil on field conditions. Five trials were done, being one trial done in laboratory, 3 in green house, and one at field condition. According to the results, the fruit fly is susceptible to the nematodes when exposed at the phases of pre-pupa and one day aged pupa. H. indica IBCB n5 was the most virulent nematode against C. capitata. At the field trial aiming to evaluate the efficiency of H. indica IBCB n5 against C. capitata and C. psidii, all the treatments with the nematode differed significantly from the control. H. indica IBCB n5 showed being much more virulent to the fruit fly, with insect mortality ranging from 66% to 93% at the dosages from 1 to 10 IJ/cm2, respective than to the guava weevil, with mortality ranging from 33 to 50%, respectively

False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. Major concerns such as progressive insecticidal resistance, the negative impact of insecticides on the environment, as well as the influence of consumers opposed to chemical residues on fruit, created opportunities for biological control methods such as Sterile Insect Technology (SIT). This technology is now established in the Western and Eastern Cape provinces of South Africa as an effective, sustainable alternative to conventional FCM control methods. Due to the prevalence of the pest in all citrus producing areas of South Africa, potential for SIT to expand is enormous. Success of an SIT programme is highly dependent on efficient application of the technology to achieve its objectives in a timeous manner. The aim of this study was to advance the application of SIT for control of FCM on citrus in South Africa, by investigating the effect of certain critical stages in the process. The effect of long-distance transportation on fitness of irradiated FCM was determined, showing reduced performance with cold-immobilized transport. A significant decrease in flight ability and longevity of irradiated FCM was found, although critically, realized fecundity was not affected. The effect of two different insecticides in the pyrethroid and organophosphate chemical groups were investigated for their residual effect on mortality of released irradiated FCM, to determine if these pest control programmes could be integrated. Both chlorpyrifos and tau-fluvalinate were effective in killing irradiated FCM for a number of days after application, after which degradation of the active ingredient rendered it harmless. This effect was found to be similar for irradiated and non-irradiated males, consequently ratios of sterile : wild male FCM should be retained regardless of whether sprays are applied or not. The modes for release of sterile FCM in an SIT programme were investigated. Efficacy of ground and aerial release platforms were tested by evaluating the recovery of released irradiated male FCM in these orchards. More irradiated FCM were recovered in orchards released from the ground compared to air. However, an economic analysis of both methods shows application of irradiated insects over a large geographical area is more cost-effective by air. Depending on the terrain and size of the target area, a combination of both methods is ideal for application of SIT for control of FCM in citrus. Development of application technology for advance of the programme is discussed and recommendations for future research and development are offered.

Inbreeding was thought to be responsible for the loss in the second filial generation (F₂) of Amalafrida leithella Dyar, Cactoblastis mundelli Heinrich, Nanaia sp. Heinrich, Sigelgaita sp. Heinrich and Sigelgaita transilis Heinrich in the laboratory. This pre-empted the investigation of the effects of inbreeding on another cactophagous moth, Mimorista pulchellalis Dyar, an established biological control agent of jointed cactus in South Africa. Initially three populations were set up. A randomly-mating control (OUT 1) population, and a sibmating experimental (IN 1) population, consisted of laboratory-reared stock . A second experimental population (KR 1) comprised a small number of field-collected randomly-mating individuals which recreated the conditions under which the five abovementioned species were lost. The inbreeding depression of fifteen fitness components was assessed. The mean values of each component in each generation of treatments IN 1 and KR 1 were compared with those of OUT 1. In addition the mean values of IN 1 were regressed against the coefficients of inbreeding since inbreeding depression is linear with respect to the probability of two genes at any locus being homozygous through ancestry. The component egg viability was important because a reduction in OUT 1, IN 1 and KR 1 in the F₂ resulted from mated females producing no viable eggs. Duplicate treatments OUT 2, IN 2 and KR 2 were set up to confirm whether this was a general F₂ phenomenon. Assessment of the fitness components prevented a direct evaluation of the numbers of offspring produced. However a hypothetical estimate of population size and growth rate was made using the percentage survival calculated from life-table analysis. Although not statistically demonstrable in the component analysis, life table analysis indicated that egg viability suffered an inbreeding depression and affected population fitness. It was also evident that treatments OUT 1 and 2 were fitter than treatments IN 1 and 2 and KR 1 and 2 with respect to population fitness. Thus, inbreeding, resulting from sibmating and introduction of a small number of individuals to a laboratorYJ caused a decrease in numbers of offspring produced and population growth rate. This is important in relation to the demise of the cactophagous Lepidoptera and to future biological control programmes.

The diamondback moth, Plutella xylostella (L.), is a pest on crucifer crops worldwide, damaging the leaves, florets and seed pods of many crucifers including cabbage, cauliflower, broccoli and canola. It has been controlled using broad-spectrum insecticides, but this has led to a rapid build-up of insecticide resistance. In the Grahamstown area of the Eastern Cape Province, South Africa, diamondback moth showed resistance to cypermethrin (a pyrethroid) on commercially grown cabbages. Therefore it is imperative that other methods of control be adopted, including both cultural control and biological control using parasitoids, and that these are incorporated into an Integrated Pest Management (IPM) programme. The diamondback moth and its parasitoids were monitored weekly from April 1997 to November 1999 at three sites near Grahamstown. One site was a commercial farm with an active insecticide spraying program; the others were unsprayed. Infestation levels were highest during spring (September to November) and autumn (March to May). Nine species of parasitoids were associated with the diamondback moth, with abundances being highest over spring and early summer (September to December). Cotesia plutellae (Kurdjumov) dominated the sprayed site, while the unsprayed sites yielded a complex of parasitoids, including C. plutellae, Diadegma mollipla (Holmgren), Diadromus collaris Gravenhorst and Oomyzus sokolowsldi (Kurdjumov). Parasitism levels ranged between 10 and 90%. There was a large amount of site-to-site and year-to-year variation. Parasitoids were an effective mortality factor against the diamondback moth. The effects of temperature on development and mortality, and of field size and non-crop plants on the distribution of diamondback moth and its parasitoids, were investigated. The results show that high temperatures can depress pest populations, and that the size and surroundings of fields can be manipulated to improve cultural control of the diamondback moth. Suggestions for effective rPM in the Eastern Cape Province include a reduction in insecticide applications, the use of bioinsecticides, for example Bacillus thuringiensis Berliner (Bt) and the encouragement of indigenous parasitoids by planting suitable nectar sources. Cultural control methods are also important and involve removal of cabbage refuse after harvest, management of wild crucifers around cabbage fields, scouting and monitoring the moth population and determining the optimal field size to assist with control by parasitoids.

Orientador: Daniela Carvalho dos Santos / Banca: Hélio Conte / Banca: Daniela Oliveira Pinheiro / Resumo: Ceraeochrysa claveri, popularmente conhecido como bicho lixeiro, é um inseto predador de ovos e larvas de insetos pragas encontrados em diferentes ecossistemas agrícolas. O bicho lixeiro se destaca por apresentar potencial para ser utilizado em programas de Controle Biológico, uma vez que é predador na fase de larva, apresenta curto tempo de desenvolvimento, fácil criação massal e alto potencial reprodutivo na fase adulta. Com o propósito de aprofundar o conhecimento na morfologia e ultraestrutura do intestino médio de C. claveri, o objetivo deste trabalho foi caracterizar histológica e ultraestruturalmente o intestino médio de C. claveri no terceiro ínstar de larva, pupa e adulto. Objetivamos, ainda, avaliar o efeito da ingestão do óleo de nim (Azadirachta indica A. Juss) (Meliaceae) sobre as células epiteliais do intestino médio de larva de terceiro ínstar, pupa e adulto de C. claveri. Ovos de Diatraea saccharalis foram tratados com óleo de nim em três diferentes concentrações (0,5; 1 e 2%) e oferecidos como alimento a larvas de C. claveri durante todo período larval. Após coleta e fixação do intestino médio de larva, pupa e adulto (grupos controle e tratados com óleo de nim), as regiões do mesêntero (anterior, média e posterior) foram processadas para análise histológica (citoquímica e imunocitoquímica) e para análise ultraestrutural. Nossos resultados mostraram que o intestino médio de C. claveri, para as três fases do ciclo de vida, é do tipo pseudoestratificado, composto por células colunares, regenerativas e endócrinas; diferenças morfológicas e ultraestruturais no epitélio permitem caracterizar duas regiões, anterior e posterior, atuando respectivamente na secreção e absorção; região média apresenta características... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Ceraeochrysa claveri, popularly known as trash-carriers is a predator insect of eggs and larvae of insect pests found in different agroecosystems. The trash-carriers stands out because it has the potential to be used in biological control programs, once it is a predator in the larval stage; it has short developmental times, easy mass-rearing and high reproductive potential in adult stage. In order to deepen the knowledge on the morphology and ultrastructure of the midgut of C. claveri, the aim of this study was to characterize histological and ultrastructurally the midgut of C. claveri in the third instar larva, pupa and adult. We also evaluate the effect of ingestion of neem oil (Azadirachta indica A. Juss) (Meliaceae) on the midgut epithelial cells of third instar larva, pupa and adult of C. claveri. Diatraea saccharalis eggs were treated with neem oil at three different concentrations (0.5, 1 and 2%) and provided as food to larvae of C. claveri throughout the larval period. After collected and fixed the midgut of larva, pupa and adult (control and treated groups with neem oil), the midgut regions (anterior, middle and posterior) were processed for histological (cytochemistry and immunocytochemistry) and for ultrastructural analysis. Our results showed that the midgut of C. claveri for the three phases of the life cycle is of pseudostratified type, consisting of columnar cells, regenerative cells, and endocrine cells, morphological and ultrastructural differences in the epithelium allowed to characterize two regions, the anterior and posterior, they working mainly in the secretion and absorption respectively; the middle region has intermediate characteristics, and it is considered a transitional area. The ingestion of neem oil in the larval stage of C. claveri showed to be toxic to this predator, causing ultrastructural alterations... (Complete abstract click electronic access below) / Mestre

Made available in DSpace on 2014-06-11T19:30:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-24Bitstream added on 2014-06-13T18:59:57Z : No. of bitstreams: 1 scudeler_el_me_botib.pdf: 6021367 bytes, checksum: 931a3db0cfc7da410cba6a737f5365bf (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Ceraeochrysa claveri, popularmente conhecido como bicho lixeiro, é um inseto predador de ovos e larvas de insetos pragas encontrados em diferentes ecossistemas agrícolas. O bicho lixeiro se destaca por apresentar potencial para ser utilizado em programas de Controle Biológico, uma vez que é predador na fase de larva, apresenta curto tempo de desenvolvimento, fácil criação massal e alto potencial reprodutivo na fase adulta. Com o propósito de aprofundar o conhecimento na morfologia e ultraestrutura do intestino médio de C. claveri, o objetivo deste trabalho foi caracterizar histológica e ultraestruturalmente o intestino médio de C. claveri no terceiro ínstar de larva, pupa e adulto. Objetivamos, ainda, avaliar o efeito da ingestão do óleo de nim (Azadirachta indica A. Juss) (Meliaceae) sobre as células epiteliais do intestino médio de larva de terceiro ínstar, pupa e adulto de C. claveri. Ovos de Diatraea saccharalis foram tratados com óleo de nim em três diferentes concentrações (0,5; 1 e 2%) e oferecidos como alimento a larvas de C. claveri durante todo período larval. Após coleta e fixação do intestino médio de larva, pupa e adulto (grupos controle e tratados com óleo de nim), as regiões do mesêntero (anterior, média e posterior) foram processadas para análise histológica (citoquímica e imunocitoquímica) e para análise ultraestrutural. Nossos resultados mostraram que o intestino médio de C. claveri, para as três fases do ciclo de vida, é do tipo pseudoestratificado, composto por células colunares, regenerativas e endócrinas; diferenças morfológicas e ultraestruturais no epitélio permitem caracterizar duas regiões, anterior e posterior, atuando respectivamente na secreção e absorção; região média apresenta características... / Ceraeochrysa claveri, popularly known as trash-carriers is a predator insect of eggs and larvae of insect pests found in different agroecosystems. The trash-carriers stands out because it has the potential to be used in biological control programs, once it is a predator in the larval stage; it has short developmental times, easy mass-rearing and high reproductive potential in adult stage. In order to deepen the knowledge on the morphology and ultrastructure of the midgut of C. claveri, the aim of this study was to characterize histological and ultrastructurally the midgut of C. claveri in the third instar larva, pupa and adult. We also evaluate the effect of ingestion of neem oil (Azadirachta indica A. Juss) (Meliaceae) on the midgut epithelial cells of third instar larva, pupa and adult of C. claveri. Diatraea saccharalis eggs were treated with neem oil at three different concentrations (0.5, 1 and 2%) and provided as food to larvae of C. claveri throughout the larval period. After collected and fixed the midgut of larva, pupa and adult (control and treated groups with neem oil), the midgut regions (anterior, middle and posterior) were processed for histological (cytochemistry and immunocytochemistry) and for ultrastructural analysis. Our results showed that the midgut of C. claveri for the three phases of the life cycle is of pseudostratified type, consisting of columnar cells, regenerative cells, and endocrine cells, morphological and ultrastructural differences in the epithelium allowed to characterize two regions, the anterior and posterior, they working mainly in the secretion and absorption respectively; the middle region has intermediate characteristics, and it is considered a transitional area. The ingestion of neem oil in the larval stage of C. claveri showed to be toxic to this predator, causing ultrastructural alterations... (Complete abstract click electronic access below)

As cultivares de algodoeiro apresentam caracterÃsticas morfolÃgicas e quÃmicas diferentes que podem influenciar a susceptibilidade Ãs pragas. Neste contexto, esse trabalho teve o objetivo de comparar a suscetibilidade das cultivares de algodoeiro herbÃceo BRS Antares, BRS 201, BRS Facual, BRS IpÃ, BRS ITA 96 BRS Sucupira, CNPA 8H, CNPA ITA 90, CNPA Precoce 3 e DeltaOpal Ãs pragas agrÃcolas, nas condiÃÃes de Fortaleza, CE. No experimento em campo, o delineamento experimental utilizado foi o de blocos casualizados, com trÃs repetiÃÃes para as plantas submetidas à infestaÃÃo natural e uma repetiÃÃo para as plantas com controle da infestaÃÃo. Cada parcela constituiu-se de trÃs linhas de trÃs metros, sendo a Ãrea Ãtil composta somente da linha central constituÃda por treze plantas. Para o monitoramento das pragas fez-se a cada sete dias observaÃÃes em campo. Na InfestaÃÃo por cochonilha (Planococcus sp.) e pulgÃo (Aphis gossypii) anotou-se o nÃmero de plantas infestadas quando essas apresentavam pelo menos uma colÃnia da espÃcie. Para o ataque de besouro-creme (Costalimaita ferruginea) anotou-se o nÃmero de plantas que apresentavam dano ≥ 10% do nÃmero total de folhas. Na infestaÃÃo por lagarta-rosada (Pectinophora gossypiella) anotou-se o nÃmero de plantas atacadas. Foram consideradas as seguintes caracterÃsticas agronÃmicas: (a) altura de inserÃÃo do primeiro ramo frutÃfero, (b) nÃmero de nÃs atà o primeiro ramo frutÃfero, (c) altura de plantas, (d) nÃmero de ramos frutÃferos por planta, (e) nÃmero de capulhos por planta, (f) dias para abertura da primeira flor, (g) dias para abertura do primeiro capulho, (h) peso mÃdio do capulho, (i) percentagem de fibra, (j) peso de 100 sementes, (l) precocidade da colheita e (m) produÃÃo. No experimento de casa-de-vegetaÃÃo, plantas das dez cultivares foram infestadas apÃs dezesseis dias de germinaÃÃo com cinco pulgÃes adultos. A contagem do nÃmero de adultos/planta foi realizada apÃs vinte e quatro horas de infestaÃÃo e a segunda avaliaÃÃo referente ao nÃmero de ninfas e de adultos/planta foi feita decorridos seis dias da instalaÃÃo do experimento. O delineamento experimental utilizado foi o inteiramente casualizado com dez tratamentos e seis repetiÃÃes. As pragas cochonilha, pulgÃo e besouro-creme nÃo demonstraram diferenÃa na preferÃncia das cultivares. A infestaÃÃo pela lagarta-rosada foi maior nas cultivares: BRS Antares, CNPA Precoce 3 e DeltaOpal. Foram encontradas reduÃÃes significativas em relaÃÃo ao controle para as seguintes caracterÃsticas: altura de inserÃÃo do primeiro ramo frutÃfero, altura das plantas, produÃÃo, precocidade e peso mÃdio dos capulhos. Considerando a infestaÃÃo das pragas e o rendimento de fibra na produÃÃo total entre as plantas infestadas, verificou-se que a cultivar CNPA Precoce 3 e CNPA ITA 90 apresentaram valores inferior e superior Ãs outras cultivares, respectivamente. Na infestaÃÃo artificial feita com os pulgÃes, nÃo houve diferenÃa significativa entre as cultivares, as quais apresentaram em mÃdia 4,06 adultos apÃs 24 horas de infestaÃÃo, 7,76 e 62,91 ninfas e adultos apÃs seis dias de infestaÃÃo, respectivamente. Pode-se concluir que a CNPA ITA 90 foi a cultivar com melhor desempenho nesta pesquisa em frente à susceptibilidade Ãs pragas. / The cotton cultivars have different morphological and chemical characteristics that may influence the susceptibility of the plants to pests. The objective of this work was to compare the susceptibility of the Upland cotton cultivars BRS Antares, BRS 201, BRS Facual, BRS IpÃ, BRS ITA 96, BRS Sucupira, CNPA 8 H, CNPA ITA 90, CNPA Precoce 3 and DeltaOpal to agricultural pests in the conditions of Fortaleza-CearÃ, Brazil. In field tests, the experimental design was a completely randomized block with three replications for the plants submitted to natural infestation and one repetition for the plants with control of the infestation. Each experimental unit had three rows with three meters long. The area observation was composed by the central row with thirteen plants. The evaluations of the infestations were made across observations in the field ever each seven days. The number of plants infested by mealybug (Planococcus sp.) and aphid (Aphis gossypii) was recorded each seven days when the plants had at least one colony of the species. The number of plants infested by yellow beetle (Costalimaita ferruginea) was recorded when the plants had at least 10% or more leaves damaged. In the infestation for pink bollworm (Pectinophora gossypiella) it was recorded the number of attacked plants. The following agronomic characteristics were considered: (a) first fruiting branch height, (b) number of nodes for the first fruiting branch, (c) plant height, (d) number of fruiting branches/plant, (e) number of boll/plant, (f) days of first flower open, (g) days of first boll open, (h) average of boll weight, (i) fiber percentage, (j) 100 seeds weight, (l) earliness and (m) yield. In greenhouse test, the plants of the ten cultivars were infested with six adult aphids after six days of germination. The number of adults/plant was recorded twenty four hours after infestation and the number of ninphs and adults/plant was also recorded six days after beginning of the experiment. The experimental design was a completely randomized with six replication of each cultivar. There were no significant differences among the cultivars damaged by Planococcus sp, A. gossypii, C. ferruginea. The cultivars BRS Antares, CNPA Precoce 3 and DeltaOpal were more infested by P. gossypiella than the other cultivars. There were significant differences among infested plants and control plants for the following characters: first fruiting branch height, plant height, earliness, yield and average of boll weight. CNPA Precoce 3 yielded less fibers and CNPA ITA 90 had the highest fiber yield. The greenhouse results indicated that there were no significant differences among the cultivars which showed an average of 4,06 adults twenty four hours after infestation and also 7,76 and 62,91 ninphs and adults respectively, six days after the infestation. CNPA ITA 90 cultivar was the one with smaller susceptibility to pests.

Made available in DSpace on 2017-07-10T19:23:54Z (GMT). No. of bitstreams: 1 Tania_ Claudia Pintro.pdf: 1091369 bytes, checksum: f8c1d3904413167a06cca540a4fcaa7a (MD5) Previous issue date: 2015-02-12 / The objective of this study was to evaluate the effect of insect infestation Sitophilus zeamais on the physicochemical and microbiological quality in wheat grains, during storage in times of 0, 45, 90 and 135 days, in three temperatures: 20, 25 and 30 ºC. For the conduct of the experiment, wheat grain samples were collected (Triticum aestivum L.) of the cultivar BRS Tanager, produced in Western Paraná, from the harvest of 2013. For storage, the grains had water contents below 13%, being stored about 400 g in sealed glass containers with fabric type voil in order to facilitate gas exchange, totaling 72 containers stored in incubator BOD the temperature of 20, 25 and 30 ° C for 135 days. Of the total of containers, 36 were infested with 20 adults of Sitophilus zeamais and the other 36 were kept without infestation. Every 45 days, were samples for subsequent chemical-physical analysis (moisture, ash, and total protein population variation of insects), microbiological (filamentous fungi count and identification of fungi producing of deoxynivalenol) and quantifying of deoxynivalenol for the three temperatures at baseline and 135 days with and without infestation. The samples were evaluated by the use of analysis of variance (ANOVA) and for test of comparison of means the Tukey test was applied, with 5% level of significance. It was observed that, with increased storage time, increased population of insects, mainly at temperatures of 20 and 25° C, reducing the value of the weight of the hectolitre at both temperatures. In general, the water content was reduced with increasing temperature and time of the storage. The contents of protein, in the treatments with and without infestation, increased with storage time. In general, the ash content increased with time of storage in grain, with and without infestation in the three temperatures evaluated. The total count of CFU of filamentous fungi was greater for grains of wheat infested. The results obtained indicate that all samples of grain stored, analyzed in the initial time and 135 days, with and without infestation, for use at temperatures of 20, 25 and 30 ᵒC, are within the maximum tolerable limit of 3000 μg of deoxynivalenol per kg of wheat. / O objetivo deste trabalho foi avaliar o efeito da infestação de insetos Sitophilus zeamais sobre a qualidade físico-química e microbiológica de grãos de trigo, durante o armazenamento nos tempos de 0, 45, 90 e 135 dias e em três, temperaturas: 20, 25 e 30 ºC. Para a condução do experimento, foram coletadas amostras de grãos de trigo (Triticum aestivum L.) do cultivar BRS Tangará, produzido na região Oeste do Paraná e provenientes da safra de 2013. Para o armazenamento, os grãos apresentaram teores de água abaixo de 13%, sendo armazenados cerca de 400 g em recipientes de vidros, fechados com tecido tipo voil a fim de facilitar as trocas gasosas, totalizando 72 recipientes armazenados em câmara incubadora B.O.D. nas temperaturas de 20, 25 e 30 °C, no tempo de 135 dias. Do total de recipientes, 36 foram infestados com 20 insetos adultos de Sitophilus zeamais e os outros 36 foram mantidos sem infestação. A cada 45 dias, foram retiradas amostras, para posterior análise físico-química, (teor de água, cinzas, proteína total e variação populacional de insetos), microbiológicas (contagem de fungos filamentosos e identificação dos fungos produtores de desoxinivalenol) e de quantificação de desoxinivalenol, para as três temperaturas no tempo inicial e de 135 dias com e sem infestação. As amostras foram avaliadas pelo emprego da análise de variância (ANOVA) e para o teste de comparação de médias foi aplicado o teste de Tukey, com nível de 5% de significância. Observou-se que conforme aumentava o tempo de armazenamento aumentava também a população de insetos, principalmente para as temperaturas de 20 e 25 °C, reduzindo o valor do peso hectolitro em ambas as temperaturas. Em geral, o teor de água diminuiu quando ocorreu aumento de temperatura e tempo de armazenamento. Os teores de proteínas, nos tratamentos com e sem infestação, aumentaram com o tempo de armazenamento. Em geral, o teor de cinzas aumentou com o tempo de armazenamento nos grãos com e sem infestação, nas três temperaturas avaliadas. A contagem total de UFC de fungos filamentosos foi maior para grãos de trigo infestados. Os resultados obtidos mostram que todas as amostras de grãos armazenados, analisadas no tempo inicial e de 135 dias, com e sem infestação para as temperaturas de 20 25 e 30 ᵒC, estão dentro do limite máximo tolerável de 3000 μg de desoxinivalenol por kg de trigo

Orientador: Maria Lígia Rodrigues Macedo / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-26T06:33:25Z (GMT). No. of bitstreams: 1 Dantzger_Miriam_D.pdf: 2351280 bytes, checksum: b2da4015dedd7611bbce146ad9143d6f (MD5) Previous issue date: 2014 / Resumo: Os inibidores de proteinases extraídos de plantas têm se mostrado promissores como um método alternativo no combate aos insetos-pragas. Neste estudo, um inibidor de peptidase foi isolado de sementes de Clitoria fairchildiana (Papilionoideae), denominado CFPI, caracterizado funcional e estruturalmente e sua atividade inseticida foi avaliada. CFPI foi purificado por exclusão molecular, seguido por coluna de interação hidrofóbica e apresentou um pico majoritário com atividade inibitória após ter sido submetido à filtração com alta resolução. Estudos cinéticos realizados com CFPI purificado mostraram uma atividade inibitória do tipo competitiva contra tripsina e quimotripsina bovinas, com uma estequiometria de inibição de 1:1 para ambas as enzimas. A constante de inibição de CFPI contra tripsina e quimotripsina bovinas foram 3,3 x 10-10 e 1,5 x 10-10 M, respectivamente, revelando uma forte capacidade de ligação. Eletroforese em SDS-Page mostrou que CFPI possui uma única cadeia polipeptídica, com uma massa molecular aparente de 15 kDa, sob condições não redutoras. Entretanto, o inibidor apresentou uma massa acurada de 7973 Daltons determinada por MALDI-TOF, sugerindo que CFPI forme dímeros em solução. Essa característica, aliada à estequiometria de inibição para tripsina e quimotripsina, à constante de inibição (Ki) para ambas as enzimas e ao sequenciamento e alinhamento N-terminal, permitiram classificar CFPI como membro da família Bowman-Birk de inibidores. O inibidor manteve-se estável ao aquecimento progressivo por 30 min a cada temperatura, variando de 37 até 100 ?C e a análise de dicroísmo não mostrou mudanças no espectro a 207 nm após aquecimento à 90 ?C e subsequente resfriamento. Além disso, CFPI mostrou atividade sobre uma ampla faixa de pH (2-10). Em contraste, a redução de CFPI com DTT resultou em perda de atividade inibitória contra tripsina e quimotripsina. CFPI exibiu atividade inibitória considerável contra enzimas tripsinas de Anagasta kuehniella (76%), Diatraea saccharalis (59%) e Heliothis virescens (49%). Suas propriedades inseticidas foram confirmadas a partir do impacto negativo causado no crescimento de A. kuehniella e C. cephalonica. O inibidor exerceu efeito antinutricional sobre A. kuehniella tanto na geração F0 como em F1 / Abstract: Proteinase inhibitors isolated from plants have shown a promising alternative method against insect pests. In this study, a proteinase inhibitor was isolated from Clitoria fairchildiana seeds (CFPI). CFPI was functional and structurally characterized and its insecticidal activity was evaluated. CFPI was purified by molecular exclusion, following by hydrophobic interaction column and showed a majoritarian peak with inhibitory activity after high resolution filtration gel column. Kinetic studies of the purified inhibitor showed a competitive¿type inhibitory activity against bovine trypsin and chymotrypsin, with an inhibition stoichiometry of 1:1 for both enzymes. The inhibition constants against trypsin and chymotrypsin were 3.3 ×10?10 and 1.5 × 10?10 M, respectively, displaying a tight binding property. SDS¿PAGE showed that CFPI has a single polypeptide chain with an apparent molecular mass of 15 kDa under non¿reducing conditions. However, MALDI¿TOF analysis demonstrated a molecular mass of 7.973 Da, suggesting that CFPI forms dimers in solution. This feature, combined with the stoichiometry of inhibition for trypsin and chymotrypsin, the inhibition constant (Ki) for both enzymes and the N-terminal sequencing, allowed classifying CFPI as a member of Bowman-Birk family inhibitors. CFPI remained stable to progressive heating for 30 min to each temperature range of 37 up to 100 °C and CD analysis exhibited no changes in spectra at 207 nm after heating at 90 °C and subsequent cooling. Moreover, CFPI was active over a wide pH range (2¿10). In contrast, reduction with DTT resulted in a loss of inhibitory activity against trypsin and chymotrypsin. CFPI also exhibited remarkable inhibitory activity against larval midgut trypsin enzymes from Anagasta kuehniella (76%), Diatraea saccharalis (59%) and Heliothis virescens (49%). Its insecticidal properties were further analysed by bioassays and confirmed by negative impact on growth of A. kuehniella and C. cephalonica. The inhibitor exhibited antinutritional effect on A. kuehniella in the F0 and F1 generations / Doutorado / Bioquimica / Doutora em Biologia Funcional e Molecular