Master of Science / Department of Entomology / Thomas W. Phillips / The rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae), is a very common worldwide pest of stored grains. The orientation of C. ferrugineus beetles to various semiochemicals was studied in both the laboratory and field. In laboratory experiments glass two-choice pitfall bioassay dishes were used. Mixed-sex populations of beetles responded positively and significantly to cucujolide I and II separately and in combination compared to controls. Bioassays using the two aggregation pheromones of the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae). Dominicalure 1 and 2 together elicited unexpected significant attractive responses by C. ferrugineus as did assays with the synthetic pheromones from other species. Bioassays showed that C. ferrugineus were not attracted to either corn meal or rolled oats, but they were attracted to commercial grain-based lures and also to ethanol, a natural fermentation product from grains. Several bioassays were conducted to test the attractiveness of naturally produced beetle volatiles to C. ferrugineus, but these showed no orientation to volatiles from either the beetles or their food only. Field tests were conducted in Kansas to test attractiveness of the synthetic pheromones cucujolide I and II in lures together with wheat versus wheat only as a control using Lindgren funnel traps. Field tests in 2014 with cucujolide II pheromone lures showed a significant response by feral C. ferrugineus when tested against wheat. Other field tests to examine dispersal of C. ferrugineus relative to grain storages showed that more C. ferrugineus were caught on the western side of grain bins compared to other cardinal directions at two different locations. Results from field and laboratory studies suggest that semiochemical-based tools can be developed to study dispersal behavior in field populations of C. ferrugineus.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2017-04-05T18:50:23Z No. of bitstreams: 1 texto completo.pdf: 1802151 bytes, checksum: 55ced28e2e4f99399e06e35026253dfd (MD5) / Made available in DSpace on 2017-04-05T18:50:23Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1802151 bytes, checksum: 55ced28e2e4f99399e06e35026253dfd (MD5) Previous issue date: 2005-06-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O interesse por métodos não-químicos de controle de pragas em grãos armazenados vem aumentando, uma vez que os consumidores estão menos tolerantes aos resíduos nos alimentos e há sucessivos relatos de resistência a inseticidas em muitas espécies de insetos de produtos armazenados. O controle biológico surge como uma das alternativas e existem vários casos de sucesso desse método, principalmente quando inserido em programas de manejo integrado de pragas. Pesquisas com o ácaro Acarophenax lacunatus (Cross & Krantz) vêm sendo desenvolvidas e demonstram que sua utilização no controle biológico de coleópteros-praga de produtos armazenados é promissora. Assim, esta pesquisa foi dividida em cinco etapas que abordam estudos básicos com este inimigo natural, visando fornecer subsídios para sua futura liberação em programas de controle biológico em ambientes de armazenamento. A primeira etapa está relacionada com a capacidade de A. lacunatus sobreviver à inanição sob diferentes temperaturas. Foram simuladas situações de escassez de alimento, sendo observado que A. lacunatus conseguiu sobreviver na ausência de alimento por até 108 horas, nas temperaturas de 20, 25 e 28 0C. A segunda etapa avaliou se este ácaro dispersa numa massa de grãos e consegue localizar o hospedeiro Rhyzopertha dominica (Fabricius). Constatou-se a habilidade de A. lacunatus em penetrar até 20 cm de profundidade e localizar ovos de R. dominica em diferentes regiões na massa de grãos, colonizando o ambiente de forma ativa. A terceira etapa consistiu no estudo da biologia de A. lacunatus sobre os hospedeiros Tribolium castaneum (Herbst) e Cryptolestes ferrugineus (Stephens) nas temperaturas 20, 25, 28, 30, 32 e 35 0C. A temperatura influenciou os parâmetros biológicos do ácaro, sendo a taxa intrínseca de crescimento (rm), a taxa líquida reprodutiva (Ro) e a razão sexual diretamente proporcionais ao aumento da temperatura. As maiores progênies (19 indivíduos/fêmea do ácaro em T. castaneum e 15 indivíduos/fêmea do ácaro em C. ferrugineus) foram observadas à temperatura de desenvolvimento. 30 A 0 C, demonstrando quarta que essa temperatura favorece seu etapa abordou um estudo de seleção de linhagens de A. lacunatus, que foram mantidas por sucessivas gerações, isoladamente, sobre os hospedeiros T. castaneum, C. ferrugineus e R. dominica. Teve como objetivo testar a preferência do ácaro por ovos desses coleópteros, em testes mensais, durante nove meses, na tentativa de detectar possíveis mudanças nessa preferência que possam indicar a seleção de linhagens desse ácaro com uma melhor performance sobre estes insetos. Houve uma influência significativa do tempo sobre a taxa de parasitismo, sendo R. dominica o hospedeiro mais parasitado em todas as situações, indicando que esta preferência parece estar fixada geneticamente, uma vez que A. lacunatus vem sendo mantido sobre este hospedeiro desde sua introdução no Brasil. Foi calculada a herdabilidade (h2) para T. castaneum e C. ferrugineus, sendo que este valor foi maior para T. castaneum. Isso indica que a seleção de linhagens desse ácaro com um melhor desempenho sobre T. castaneum pode ser realizada. Já a quinta etapa consistiu no estudo do potencial de parasitismo de A. lacunatus sobre o coleóptero T. castaneum, onde observou-se que o aumento da densidade do ácaro implicou no aumento do número de ovos parasitados, com conseqüente redução das populações desse inseto. Assim, novas informações sobre A. lacunatus são acrescentadas e indicam que sua utilização no controle biológico de T. castaneum, C. ferrugineus e R. dominica pode ser viável. / The interest for non-chemical control methods of stored grain pests is increasing since the consumers are less tolerant about insecticide residues on food and there are reports of insecticide resistance in insects of stored products. The biological control appears as an alternative and there are several success cases of the control method, mainly within integrated pest management programs. Studies with the mite Acarophenax lacunatus (Cross & Krantz) have been carried out and show the potential of its use in the biological control of stored grain beetles. Therefore the present study was divided in five phases encompassing basic investigations with this natural enemy aiming to provide support for its future use in biological control programs in storage facilities. The first phase is related with the capacity of A. lacunatus to survive under starvation at different temperatures. This mite species was able to survive for up to 108 hours at the temperatures of 20, 25 and 28 0C. The second phase focused on assessing the dispersal of this mite species in the grain mass and its ability to locate its host Rhyzopertha dominica (Fabricius). The mite was able to penetrate up to 20 cm deep and locate eggs of R. dominica in different regions of the grain mass efectivelly colonizing the environment. The third phase involved the biology of A. lacunatus on the hosts Tribolium castaneum (Herbst) and Cryptolestes ferrugineus (Stephens) at the temperatures 20, 25, 28, 30, 32 and 35 0C. The temperature affected the biological parameters of the mite with a proportional increase in the intrinsic growth rate (rm) net reproductive rate (Ro) and sex ratio with increased temperature. The greates progenies (19 individuals/female in T. castaneum and 15 individuals/female in C. ferrugineus) were observed at 30 0C temperature showing that this temperature favors the mite development. The fourth phase focused the selection of mite strains maintained for several generations in different hosts (T. castaneum, C. ferrugineus and R. dominica). The objective was to test the host egg preference of each selection line in monthly tests trying to detect changes in preference through time. There was a significant influence of time on the parasitism rate with R. dominica always being the preferred host and indicating that this preference was genetically fixed in the mite population. The heritability (h2) of egg preference for the hosts T. castaneum and C. ferrugineus was calculated and higher values were observed for T. castaneum. This indicates that selection for better performance in T. castaneum is possible. In the fifth phase of the present effort, we investigated the parasitism potential of A. lacunatus in the beetle T. castaneum where an increase in mite density led to an increase in the number of parasitized eggs with a consequent reduction of this insect population. Therefore, new information on A. lacunatus was obtained indicating the viability of its use in biological control programs against T. castaneum, C. ferrugineus and R. dominica.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-06-27T17:46:33Z No. of bitstreams: 1 texto completo.pdf: 647098 bytes, checksum: b268e217f616628b33c7d54bc5890149 (MD5) / Made available in DSpace on 2017-06-27T17:46:33Z (GMT). No. of bitstreams: 1 texto completo.pdf: 647098 bytes, checksum: b268e217f616628b33c7d54bc5890149 (MD5) Previous issue date: 2001-07-24 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Com a necessidade de buscar alternativas para o uso de agentes químicos em produtos armazenados, devido ao desenvolvimento de resistência em insetos-praga e à possibilidade de contaminação dos alimentos por resíduos, os programas de erradicação de pragas têm dado maior ênfase ao controle biológico. Em razão do exposto, este trabalho foi dividido em três etapas que abordaram o potencial de parasitismo do ácaro Acarophenax lacunatus (Prostigmata: Acarophenacidae) em cinco espécies de insetos de produtos armazenados. Na primeira etapa, avaliou-se a preferência deste ácaro por ovos dos insetos Rhyzopertha dominica (Coleoptera: Bostrichidae), Dinoderus minutus (Coleoptera: Bostrichidae), Tribolium castaneum (Coleoptera: Tenebrionidae) e Oryzaephilus surinamensis (Coleoptera: Silvanidae). A segunda etapa abordou o estudo do potencial de parasitismo do ácaro em D. minutus em mandioca seca armazenada, e a terceira etapa em R. dominica, T. castaneu m, O. surinamensis e Cryptolestes ferrugineus (Coleoptera: Cucujidae) em trigo parcialmente triturado. Todos os testes foram realizados a 28 o C e escotofase de 24 horas, em câmaras climáticas do tipo B.O.D. O estudo da preferência por ovos foi realizado em dois testes: um sem chance de escolha e outro com chance de escolha, utilizando-se placas de Petri (5 cm de diâmetro), as quais foram divididas em quatro quadrantes. No teste sem chance de escolha, foram colocados, em cada quadrante, cinco ovos de apenas um dos insetos e, no centro da placa, uma fêmea de A. lacunatus em processo de fisogastria. O experimento foi realizado com cada uma das espécies. No teste com chance de escolha, foram utilizados ovos de todas as espécies, que foram distribuídos de forma que cada quadrante só recebesse ovos de um dos insetos. Da mesma forma, no centro da placa foi liberada uma fêmea fisogástrica do ácaro. Em nenhum dos testes o ácaro A. lacunatus parasitou ovos de O. surinamensis. No teste sem chance de escolha não foi observada preferência de A. lacunatus por ovos de nenhum dos outros hospedeiros. Entretanto, no teste com livre escolha, A. lacunatus parasitou mais ovos de R. dominica e D. minutus que de T. castaneum. Nas outras etapas do trabalho, avaliou-se a população final dos insetos (larvas e adultos), o número de ovos parasitados e de ácaros em processo de fisogastria, a taxa instantânea de crescimento (r i ) de A. lacunatus e dos insetos, e a perda de peso dos produtos (mandioca e trigo). Foi observado que o ácaro apresenta potencial biótico maior que o dos hospedeiros R. dominica, D. minutus e T. castaneum. Desta forma, o número de larvas e adultos destes insetos sofreu reduções em relação à testemunha, o que também ocorreu com as perdas de matéria seca ocasionadas pelo ataque dos insetos. Também não foi observado parasitismo em populações do coleóptero O. surinamensis. Pelo número de ovos parasitados de R. dominica, D. minutus, T. castaneum e C. ferrugineus, conclui-se que o ácaro A. lacunatus é um inimigo natural eficaz destes insetos, não sendo, portanto, um parasita específico de R. dominica , como se imaginava. / With the necessity of searching for alternatives to the use of chemical control agents in stored products due to development of insecticide resistance in insect-pests and the possibility of foodstuff contamination by residue, pest eradication programs have been putting more emphasis on biological control. As a consequence, the present work was divided in three phases which aimed to assess the parasitism potential of the mite Acarophenax lacunatus (Prostigmata: Acarophenacidae) in five species of insects of stored products. In the first phase, the mite preference for eggs of Rhyzopertha dominica (Coleoptera: Bostrichidae), Dinoderus minutus (Coleoptera: Bostrichidae), Tribolium castaneum (Coleoptera: Tenebrionidae) and Oryzaephilus surinamensis (Coleoptera: Silvanidae) was assessed. In the second phase, the parasitism potential of the mote species of D. minutus attacking dried stored cassava was evaluated and in the third phase the parasitism potential of the same parasitic species was assessed in R. dominica, T. castaneum, O. surinamensis e Cryptolestes ferrugineus (Coleoptera: Cucujidae) infesting wheat. All of the experiments were carried out under controlled conditions of 28 o C and 24 hours scotophase in climatic chambers of the B.O.D. type. The study of egg preference was carried out in two tests: one without opportunity of choice and other with it using Petri dishes divided in four areas. On the non-choice test five eggs of only one species was placed in each different area and a physogastric female of A. lacunatus was placed in the centre of the Petri dish (5 cm diameter). On the choice test there was only eggs of a single species in each of the four areas within the Petri dish and the same procedure was used for each potential prey species. The mite A. lacunatus did not attack eggs of O. surinamensis in any of the tests. No preference of A. lacunatus by eggs of any host was observed on the non-choice test. However, A. lacunatus attacked more eggs R. dominica and D. minutus than of T. castaneum. In the other phases of the work the final insect population (adults and larvae), egg parasitism, number of mites in physogastry, the instantaneous rate of increase of the mite and the insect populations (r i ) and weight loss of the stored product (wheat or cassava). It was observed that the mite has higher biotic potential than the hosts R. dominica, D. minutus and T. castaneum. Therefore, the number of larvae and adults of these insect species were lower than in the control treatment, what was also observed with the losses in dried matter caused by insect infestation. Again, no parasitism on eggs of O. surinamensis was observed. Based on the egg parasitism of R. dominica, D. minutus, T. castaneum and C. ferrugineus, the mite A. lacunatus is an efficient natural enemy for all of this species and not only to R. dominica, as initially reported. / Dissertação importada do Alexandria

Algunas plagas causan daños importantes en productos y granos almacenados, lo cual conlleva consecuentemente a pérdidas de producción y calidad en estos productos.Las principales plagas del arroz almacenado en España son, Sitophilus oryzae L. (Coleoptera Curculionidae), Rhyzopertha dominica Fabricius (Coleoptera: Bostrichidae), y Cryptolestes pusillus Schönherr (Coleoptera: Cucujidae). Las dos primeras son plagas primarias que atacan directamente el grano y resultan bastante destructivas debido a que sus larvas se alimentan y desarrollan dentro de él. C.pusillus es, sin embargo, una plaga secundaria que se beneficia de granos que ya están dañados y rotos.Actualmente, el uso de fumigantes e insecticidas de síntesis sigue siendo el principal método de lucha para controlar las plagas de almacén, si bien recientemente (dadas las continuas restricciones al uso de agroquímicos) existe un gran interés en la utilización de otras alternativas tales como el control biológico, el almacenamiento a bajas temperaturas, o los tratamientos con calor entre otros.Igualmente muchos productos obtenidos principalmente de plantas y que derivan del metabolismo secundario de las mismas ofrecen una fuente de bioinsecticidas que podrían representar una alternativa ecológica frente a los insecticidas de síntesis ya que su uso masivo e indiscriminado ha ocasionado problemas tales como la aparición de resistencias en determinadas especies de insectos frente a diferentes materias activas, desequilibrios ecológicos y problemas medioambientales sin olvidar el riesgo que entrañan para la salud humana.Con este trabajo se ha pretendido estudiar la actividad plaguicida de los aceites esenciales extraídos de tres plantas: Coriandrum sativum L. (Umbelliferae), Carum carvii L. (Umbelliferae) y Ocimum basilicum L. (Labiatae) y su posterior fraccionamiento para identificar dentro de estos aceites los compuestos químicos responsables de esta actividad insecticida sobre tres plagas de almacén de arroz (S. oryzae, R.dominica y C.pusillus).Del estudio de estos monoterpenoides, linalol, S-carvona y estragol resultaron tener una alta actividad insecticida sobre estas plagas. Sin embargo el E-anetol fue más selectivo para R.dominica y C.pusillus, así como el limoneno, γ-terpineno, geraniol y eucaliptol sólo resultaron activos frente a C.pusillus.Algunos monoterpenoides podrían actuar de sinergistas potenciando la actividad de otros, como podría ser el caso del alcanfor, acetato de geranilo y E-anetol con linalol en R.dominica y C.pusillus, o el caso del metoxicinamaldehido, p-anisaldehido y linalol que pueden tener efecto sinergista sobre el estragol.Este trabajo también abarcó el estudio de un posible modo de acción de estos monoterpenoides, la inhibición de la acetilcolinesterasa, para alcanzar un mayor entendimiento del comportamiento de estas sustancias en el interior del insecto.Así se pudo observar como la mayoría de monoterpenoides estudiados inhibían en cierta medida esta enzima, siendo fenchona, S-carvona y linalol los monoterpenoides que mayor inhibición originaron.Del mismo modo se observó como fenchona, γ-terpineno, geraniol y linalol inhibían competitivamente la acetilcolinesterasa, mientras que S-carvona, estragol y alcanfor producían una inhibición mixta para esta enzima.Sin embargo no se observó inhibición de la acetilcolinesterasa por parte del E-anetol a las concentraciones de monoterpenoides ensayadas.Para completar este trabajo se examinó de igual forma la capacidad que tenían estos bioinsecticidas de generar resistencia en estas tres plagas así como el mecanismo de resistencia implicado en el desarrollo de este proceso. Para ello se fueron seleccionando las poblaciones de insectos mediante la aplicación de los diferentes monoterpenoides a dosis crecientes durante 7 generaciones. De este modo se pudo calcular el factor de resistencia en cada una de las plagas y para cada uno de los monoterpenoides comparando las concentraciones letales 50 de las poblaciones seleccionadas con las poblaciones iniciales (sensibles).A continuación se analizaron tres posibles sistemas de detoxificación enzimáticos gracias al uso de sinergistas para estudiar el mecanismo de resistencia que podría estar involucrado.De tal forma se observó como estos monoterpenoides inducían lentamente resistencias resultando ventajosos en un futuro para el control de estas plagas.Esta Tesis ha englobado un estudio íntegro y profundo de estos insecticidas ecológicos desde la extracción de los aceites esenciales, seguido de la identificación de compuestos puros (CG-EM) con actividad insecticida junto con el estudio de un modo de acción de estos insecticidas, la selección de resistencia en las poblaciones de insectos y los posibles mecanismos de resistencia que pudieran estar implicados en este proceso. / Some pests cause serious damage to stored grains and stored products and consequently production and quality losses in these products.The rice weevil, Sitophilus oryzae L. (Coleoptera: Curculionidae), the lesser grain borer, Rhyzopertha dominica Fabricius (Coleoptera: Bostrichidae) and Cryptolestes pusillus Schönherr (Coleoptera: Cucujidae) were the main damaging pests found in stored rice in Spain.S.oryzae and R.dominica are primary pests attacking directly the intact grain and are quite destructive because their larvi feed and develop inside the grain whereas C.pusillus is a secondary pest which benefits from grains previously damaged.At the present time, organic synthetic pesticides are still the main method to control stored grain pests, however, recently (due to restriction in agrochemicals use) there is a great interest in using other altenatives such as biological control, storage at low-temperatures, or heat treatment.Likewise, many products obtained mainly from plants and derived from secondary metabolism have insecticidal activity against insects, such as monoterpenoids, which present a broad variety of bioinsecticide products which could turn out to be an ecologic alternative to synthetic pesticides since the majority of alternative products are not harmful for the human healthy and they become less environmentally damaging, exhibiting a low impact on the environment.In addition, it cannot be ignored the different difficulties related to resistance due to several active compounds from organic pesticides found in some species of insects.With this work, we have considered remarkable to study the insecticide activity of essential oils extracted from three plants: Coriandrum sativum L. (Umbelliferae), Carum carvii L. (Umbelliferae) and Ocimum basilicum L. (Labiatae) and carry out a bioassay-guided fractionation of their essential oils to identify which compounds were responsible for the volatile toxicity shown on three stored rice pests (S. oryzae, R. dominica and C. pusillus).Linalool, S-carvone and estragole turned out to have a high insecticide activity on these pests. Nevertheless E-anetol was more selective to R. dominica and C. pusillus, being only active on C. pusillus limonene, γ-terpinene, geraniol and eucalyptol.Some monoterpenoids were found as synergists, increasing the activity of the other ones, like for instance, camphor, geranyl acetate and E-anethole with linalool in R.dominica and C.pusillus or metoxycinnamaldehyde, p-anysaldehyde and linalool which could activate to estragole.In this work, the inhibition of acetylcholinesterase as a posible mode of action was studied as well, to reach a clear understanding about the action of these products inside the insects.The majority of monoterpenoids inhibited the enzyme acetylcholinesterase being fenchone, S-carvone and linalool the monoterpenoids that produced a higher inhibition.Furthermore, it was observed how fenchone, γ-terpinene, geraniol and linalool showed a competitive inhibition whereas S-carvone, estragole and camphor produced a mixed inhibition for this enzyme. However the enzyme acetylcholinesterase was not inhibited by E-anethole.To finish up this work, the selection for monoterpenoid resistance on these pests as well as the metabolic mechanisms implicated was studied.The resistant strains were selected from susceptible insect populations and survivors were reared separately for each monoterpenoid and successive generations were treated with higher concentrations. These populations were selected until seven times.As a result we could calculate the resistance factor on each pest (comparing lethal concentration 50 values of susceptible and resistant strains).Next, three enzymatic systems detoxifying these monoterpenoids were analysed to study the metabolic mechanism implicated.In this way we could realize that all monoterpenoids induced resistance slowly, concluding that these pesticides will be appropriated to control these pests in the future.This Thesis has concerned a study in depth about ecological insecticides from extraction of essential oils, identification of compounds (GC-MS) with insecticide activity, mode of action and study of resistance and mechanism of insecticide resistance involved in this process.