Master of Science / Department of Entomology / Frank H. Arthur / Kun Yan Zhu / Experiments were conducted to assess the effects of direct and indirect exposure scenarios, different degrees of residual flour, open and obstructed positions, and seasonal temperature variations on the efficacy of synergized pyrethrin against the red flour beetle, Tribolium castaneum (Herbst) and the confused flour beetle, Tribolium confusum Jacquelin du Val. To evaluate effects of direct and indirect exposures of T. castaneum and T. confusum eggs, larvae, pupae, or eggs to the insecticide aerosol within a flour mill, the following treatments were made to each life stage: insects treated with aerosol and transferred to treated or untreated flour, untreated insects transferred to treated flour, and insects and flour combined and treated together. Different degrees of harborage or sanitation levels were created by exposing T. confusum larvae, pupae, and adults to pyrethrin aerosol in Petri dishes containing 0, 0.1, 1, 5, and 10 g of wheat flour. Effects of pyrethrin dispersal in open and obstructed positions and seasonal temperature variations were assessed by exposing T. confusum pupae and adults in open positions and inside wooden boxes (1 m long, 20 cm wide, and 5, 10, or 20 cm high) inside experimental sheds maintained at target temperatures of 22, 27, and 32 °C. Results showed that when T. castaneum and T. confusum were directly exposed to aerosol without the flour source, or with a low amount of flour at open exposed areas, the aerosol provided good control against all life stages of T. castaneum and T. confusum. However, when insects were indirectly exposed (treated together with flour or untreated insects were transferred to treated flour), or treated together with deeper flour amounts, and exposed inside the boxes, the efficacy was greatly reduced. Eggs and pupae of both the species were more susceptible compared to larvae and adults. Additionally, the moribund adults initially observed in indirect exposure treatments, or at the deeper flour depth and exposure positions insides the boxes, were better able to recover. Generally, temperatures in the range of 22-32 °C had no significant effects on overall efficacy of pyrethrin aerosol.

Made available in DSpace on 2015-03-26T13:23:50Z (GMT). No. of bitstreams: 1 texto completo.pdf: 269289 bytes, checksum: fda931f23e88417c20515b21a0ea64cb (MD5) Previous issue date: 2005-02-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The current work aimed to assess the susceptibility of adults of Sitophilus zeamais (Motschulsky) (Coleoptera: Curculionidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), subjected to ozone treatment at different depths in the grain mass, thus estimating the lethal time for 50% and 95% of each species population, and also assess the physiological quality of the maize grains subjected to the ozone treatment. Maize grains with moisture level around 13% (wb) were distributed in cylindrical PVC containers with 20 cm diameter, 100 cm high and connections for gas injection and exhaustion. At 10 cm from the base of the container, a metallic net was placed to sustain the grain and to form a plenum for better gas distribution. Maize grains infested with adults of Sitophilus zeamais and Tribolium castaneum, obtained from laboratory colonies from climatic chamber of the B.O.D. type, were placed in cages of 3.0 cm high and 15.0 cm diameter with top and bottom of voil to evaluate the efficiency of ozone as fumigant. These cages were placed at different grain mass depths (over the plenum, middle and top of grain column) and subjected to a modified atmosphere of 50 ppm ozone under different exposure periods. The ozone was injected in a continuous flow of 8.0 L min-1 in connection located at the base (plenum) of the container. Tests of electrical conductivity, germination potential and humidity level were carried out in the maize grains to assess the ozone effect on them. The grain mass temperature was maintained around 25 oC throughout the experiment. To do that, a temperature controlled chamber was built where the containers were placed. The temperature was monitored through a data acquisition and store system called f-wire. Six cylindrical containers were used in all of the assays and in three of these ozone was injected, while the remaining ones were injected with atmospheric air (control). It was concluded that in general the increase in exposure period increased the efficiency of the treatments with 50 ppm ozone for adults S. zeamais and T. castaneum. The species S. zeamais was more susceptible. The highest exposure period to control 95% of the insects was of 240.75 h for S. zeamais and 390.18 h for T. castaneum. The greatest exposure period to control 50% of the adult insects was 124.20 h for S. zeamais and 234.75 h for T. castaneum. In general, the treatments with modified atmosphere containing 50 ppm ozone and atmospheric air did not affect the physiological quality of the maize grains. / O presente trabalho teve por meta avaliar a suscetibilidade dos adultos de Sitophilus zeamais (Motschulsky) (Coleoptera: Curculionidae) e Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), submetidos ao tratamento com ozônio em diferentes camadas da massa de grãos, estimando-se, assim, os tempos letais (TL) para 50% e 95% da população de cada espécie e, ainda, avaliar a qualidade fisiológica dos grãos de milho submetidos aos tratamentos com ozônio. Grãos de milho com teor de umidade em torno de 13% (b.u.) foram distribuídos em recipientes cilíndricos, construídos em PVC, com 20 cm de diâmetro, 100 cm de altura e conexões para injeção e exaustão de gás. A 10 cm da base do recipiente, colocou-se uma tela metálica para sustentação dos grãos e formação de um plenum para melhor distribuição do gás. Para avaliar a eficácia do ozônio como fumigante, grãos de milho infestados com adultos de Sitophilus zeamais e Tribolium castaneum, obtidos de criação contínua em câmara climática do tipo B.O.D., foram distribuídos em gaiolas de 3,0 cm de altura e 15,0 cm de diâmetro, também em PVC, sendo o fundo e a tampa confeccionados em tecido do tipo organza. Estas gaiolas foram dispostas em diferentes camadas da massa de grãos (sobre o plenum, mediana e superior) e submetidas a uma atmosfera modificada com 50 ppm de ozônio em diferentes períodos de exposição. O ozônio foi injetado em fluxo contínuo de 8,0 L min-1, em conexão localizada na base (plenum) do recipiente. Para avaliar o efeito da fumigação com o ozônio na qualidade do milho, foram realizados testes de condutividade elétrica, potencial de germinação e teor de umidade. Em todo o experimento, a temperatura da massa de grãos foi mantida próxima de 25 oC. Para tanto, construiu-se uma câmara com controle de temperatura onde foram acondicionados os recipientes cilíndricos, sendo esta monitorada por meio de um sistema de aquisição e armazenamento de dados denominado 1-wire. Em todos os ensaios foram utilizados seis recipientes cilíndricos, sendo que em três destes injetou-se ozônio e nos outros foi injetado ar atmosférico (testemunha). Concluiu-se que, em geral, o aumento do período de exposição resultou no aumento da eficácia dos tratamentos com 50 ppm de ozônio para os adultos de S. zeamais e T. castaneum. A espécie que se mostrou mais susceptível foi S. zeamais. O maior período de exposição para o controle de 95% dos insetos foi de 240,75 h para o S. zeamais e de 390,18 h para o T. castaneum. O maior período de exposição para o controle de 50% dos insetos adultos foi de 124,20 h para o S. zeamais e de 234,75 h para o T. castaneum. Em geral, os tratamentos com atmosfera modificada com 50 ppm de ozônio e com ar atmosférico, não afetaram a qualidade fisiológica dos grãos de milho.

No description available.

No description available.

No description available.

Made available in DSpace on 2015-03-26T13:30:20Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1603340 bytes, checksum: 135e2fd308ff1af60f963623548c94b3 (MD5) Previous issue date: 2006-02-17 / Empresa Brasileira de Pesquisa Agropecuária / The resistance of the insect-pests to fumigants has been frequently reported in stored grains and a present is one of the great obstacles to the control programs involving the use of chemical pesticides. This study was therefore carried out to detect phosphine resistance in populations of Tribolium castaneum, Rhyzopertha dominica and Oryzaephilus surinamensis and to recognize the existence of adaptative disadvantages of these insects in the absence of phosphine, by correlating the instantaneous rate of population growth (ri) with the resistance levels observed in different populations of the species under study. Twelve distinct populations of T. castaneum, ten of R. dominica, and eight of O. surinamensis collected in the states of Goiás, Mato Grosso, Minas Gerais and São Paulo were surveyed. The bioassays for the detection of phosphine resistance were carried out following the standard method recommended by the Food and Agriculture Organization of the United Nations (FAO), which it is based on the exposure of 50 non-sexed adult insects, in four replicates, to the discriminating concentration (DC) of the fumigant for a 20 hourperiod. Later, the concentration-response bioassays were carried out generating the concentration-mortality curves that were used to estimate LC50 and LC95 of the populations pertaining to those three species. The LC50 were used to discriminate the standard population for susceptibility in each species. The measurements of the body mass, CO2 production, and instantaneous rate of population increase (ri) in each population of each species were correlated to the resistance ratio at the LD50. The resistance ratio in T. castaneum ranged from 1,0 to 186,2-fold, whereas in R. dominica it ranged from 2,0 to 71,0-fold, and the lowest one from 1,9 to 32,2-fold for O. surinamensis. The results obtained in the resistance detection bioassays pointed out ten populations of T. castaneum, nine of R. dominica and seven of O. surinamensis that were resistant to phosphine. The relationship between the body mass and CO2 production was positive and significant indicating an increase in CO2 production with the individual body mass. The same effect is observed in the interaction between the instantaneous rate of population increase (ri) and body mass. In addition, the populations with lower CO2 production showed higher resistance ratio (RR) for all species, what is related to the phosphine resistance mechanism. The populations with higher ri showed lower resistance ratio (RR). This result indicates a worse reproductive performance of the resistant populations compared with the susceptible ones. Thus, the management strategies that are based on the interruption of the phosphine fumigation over determined time for the reestablishment of the susceptibility and eventual reintroduction of the fumigant shows a good perspective as a management tactic since the resistant populations under study showed adaptative disadvantage in relation to the susceptible ones in the absence of phosphine. / A resistência a inseticidas fumigantes em insetos-praga de grãos armazenados tem sido freqüentemente registrada e, atualmente, tem se tornado um dos grandes obstáculos aos programas de controle envolvendo o uso de produtos químicos. Com isso, objetivou-se neste estudo detectar populações de Tribolium castaneum, Rhyzopertha dominica e Oryzaephilus surinamensis resistentes à fosfina, além de aferir a existência de desvantagens adaptativas destas, na ausência da fosfina, correlacionando-se produção de CO2, massa corpórea e taxa instantânea de crescimento populacional (ri), com os níveis de resistência observados nas diferentes populações das três espécies em estudo. Foram utilizadas 12 populações distintas de T. castaneum, 10 de R. dominica e oito populações de O. surinamensis, coletadas nos Estados de Goiás, Mato Grosso, Minas Gerais e São Paulo. Os bioensaios para detecção de resistência foram conduzidos conforme método-padrão, recomendado pela Food and Agriculture Organization of the United Nations (FAO), que é baseado na exposição de 50 insetos adultos, em quatro repetições, por um período de 20 horas à concentração discriminante (CD) do gás fumigante. Posteriormente, foram conduzidos bioensaios de concentração-resposta gerando, assim, as curvas de concentração-mortalidade que foram utilizados para estimar as CL50 e CL95 das populações das três espécies. As CL50, por sua vez, foram utilizadas para discriminar a população padrão de suscetibilidade de cada espécie e o nível de resistência das demais populações. A mensuração da massa corpórea, produção de CO2 e taxa instantânea de crescimento (ri) de cada população, das três espécies avaliadas, foram correlacionados à razão de resistência para CL50. A razão de resistência para CL50 em T. castaneum variou de 1,0 a 186,2 vezes, enquanto em R. dominica foi de 2,0 a 71,0 vezes e em O. surinamensis observou-se a menor variação, de 1,9 a 32,2 vezes. Os resultados obtidos nos bioensaios de detecção de resistência indicaram 10 populações de T. castaneum, nove de R. dominica e sete de O. surinamensis resistentes a fosfina. A relação entre massa corpórea e produção de CO2 foi significativa, indicando aumento na produção de CO2 com o incremento da massa corpórea dos indivíduos, nas três espécies avaliadas. O mesmo efeito é observado na interação entre taxa instantânea de crescimento populacional (ri) e a massa corpórea. As populações com menor produção de CO2 apresentaram maior razão de resistência (RR) para CL50, para as três espécies avaliadas, fato que está relacionado ao mecanismo de resistência dos insetos à fosfina. As populações com maiores valores médios de ri apresentaram menor razão de resistência (RR) para CL50, nas três espécies avaliadas. Este resultado indica pior desempenho reprodutivo das populações resistentes, em relação às populações suscetíveis na ausência do fumigante. Portanto, estratégias de manejo da resistência que são baseadas na interrupção da fumigação com fosfina, por determinado tempo visando o restabelecimento da suscetibilidade e eventual reintrodução do fumigante apresentam boa perspectiva de sucesso. Isto porque, as populações resistentes avaliadas neste estudo apresentaram desvantagens adaptativas, em relação às suscetíveis, na ausência da fosfina.

Doctor of Philosophy / Department of Entomology / James F. Campbell / Phillip E. Sloderbeck / Robert "Jeff" J. Whitworth / Knowledge of factors influencing the establishment, persistence and distribution of stored-product pests aids the development of effective Integrated Pest Management (IPM) programs in food storage and processing facilities. This research focused primarily on Tribolium castaneum, which is one of the most important pests of mills. Populations of T. castaneum from different food facilities can potentially be interconnected by either their own dispersal behavior or by human transportation. Population genetic structure analyses based on microsatellites and other insertion-deletion polymorphisms (“indels”) showed that populations from different mills around the US are genetically distinct from each other, but the level of differentiation was not correlated with the geographic distance. A potential source of insect infestation within a food facility is spillage that accumulates outside or movement from bulk storage facilities on site. Results from three facilities showed that most stored-product species were captured both inside and outside buildings, but T. castaneum was rarely captured outside of the facilities. Spatial distribution of all species outside was associated with the proximity of buildings, not necessarily with areas with accumulated spillage. T. castaneum populations inside facilities are potentially exposed to frequent genetic bottlenecks resulting from structural fumigations. Changes in allele frequencies through time, based on the analysis of microsatellites and other indels in individuals collected in a mill, confirmed bottleneck effects. To understand how spatial distribution of T. castaneum within a mill could be influenced by environmental and physical factors, a range of variables were measured at each trap location. There was significant variation among trap locations regarding beetle captures and the variables measured, but increase in beetle captures correlated only with increase in temperature and spillage production. Tribolium castaneum response to visual cues could influence attraction to pheromone and kairomone olfactory cues used in traps. Results of laboratory experiments showed that adults respond to tall narrow black shapes and placing traps in front of these shapes can increase captures. This research provides new insights into factors influencing the spatial distribution of T. castaneum and could help in improving monitoring programs for this important pest of the food industry.

Doctor of Philosophy / Department of Entomology / David C. Margolies and Yoonseong Park / Organisms are capable of sensing environmental conditions through diverse mechanisms. Transient receptor potential channels (TRPs) are a cation channel family that has been found to function in diverse sensing mechanisms. In this dissertation, I identified the function of several TRPs in thermosensing and mechanosensing in the red flour beetle, Tribolium castaneum. Candidate TRPs were chosen based on homology to TRPs found and studied in Drosophila melanogaster. To identify the function of candidate TRPs in T. castaneum, I suppressed the expression of target genes by RNA interference technique and investigated the phenotype of each treated beetle. Temperature is a major limiting environmental factor for organisms. I tested the function of candidate TRPs in thermotaxis (behavior) and thermal acclimation (physiology). Using bioinformatics approaches, I identified three candidate TRPs – painless, pyrexia, and trpA1 – involved in high temperature sensing. To test thermotactic behavior, I investigated beetle movement on a temperature arena with two separate temperature zones. Thermal acclimation was tested by pre-exposing beetles to either 42 °C for 10 min. When treated with double stranded RNA of TRPA1 (dstrpA1), the thermotactic response of beetles at 39 and 42 °C was reduced when compared to control groups. With pre-exposure at 42 °C, survivorship of dstrpA1-treated beetles significantly increased after one minute exposure at 52 °C compared to beetles that were not pre-exposed. With dspainless treatment, beetles showed lower response to thermal acclimation and lower long-term survivorship. Beetles treated with dspyrexia showed lower recovery after heat treatment without pre-exposure at 42 °C. To identify the function of candidate TRPs in mechanosensing, I evaluated dsRNA treated beetles for survival, walking behavior, and tonic immobility. Treatment with dsnompC and dstrpA5 resulted in failure in eclosion, causing 93 % mortality in both treatments. Survivors in dsnompC showed defects in elytra sclerotization. In dsnanchung and dsinactive treatments, adults showed abnormal walking behavior and reduced walking speed that were likely caused by defects of mechanosensing in folding of the joint between the femur and tibia. For tonic immobility, beetles with dsnanchung, dsinactive, dswaterwitch and dsick2 (insect cytokine 2) treatments showed increased sensitivity to mechanical stimulation leading to tonic immobility.

Master of Science / Department of Grain Science and Industry / Subramanyam Bhadriraju / During heat treatment, the ambient temperature of grain-processing facilities is raised to 50-60°C for at least 24 hours to manage stored-product insects. Young larvae (first instars) of the red flour beetle, Tribolium castaneum (Herbst), are the most heat tolerant stage at 50-60°C. A thermal death kinetic (TDK) model predicted survival of T. castaneum young larvae exposed to six constant elevated temperatures between 42 and 60ºC. The model is based on logarithmic survival of T. castaneum as a function of time and logarithmic reduction in larval survival as a function of temperature. The model was validated with 12 independent temperature datasets collected during heat treatments of pilot-scale and commercial grain-processing facilities. Young larval survival in plastic boxes/vials with flour was used to validate model predictions. The heating rate to 50°C from the ambient among the 12 datasets ranged from 0.9-7.8°C/h. Mean absolute deviations between observed and predicted larval survival for 10 of the 12 datasets ranged from 2.1-11.4%; it was 16.2 and 18.3% for two other datasets. The TDK model can be used to predict survival of young larvae of T. castaneum based on time-dependent temperature profile obtained at any given location during heat treatment of grain-processing facilities. In three commercial grain-processing facilities heat treatments were conducted for 24-27.7 hours using forced-air gas heaters. Temperatures attained and survival of 20 eggs, 20 young larvae, and 20 adults of T. castaneum in bioassay vials at various locations were determined. Across all three facilities, 5 out of 2720 adults in 136 vials, 1 out of 960 young larvae in 48 vials, and 0 out of 1760 eggs in 88 vials were alive at the end of the heat treatment. In each facility, the time in hours for 1% predicted survival of T. castaneum young larvae was positively related to how quickly temperatures reached 50°C, and negatively related to rate of heating to 50°C from the ambient, time above 50°C in hours, and the maximum temperature. Bioassays with T. castaneum life stages and the TDK model can be used to gauge effectiveness of facility heat treatments.

Master of Science / Department of Entomology / Franklin Arthur / Kun Yan Zhu / Tribolium spp. are major pests in structures used for the processing and storage of grain-based products (e.g., flourmills, warehouses, retail stores). Consumers and regulators have little tolerance for insect-damaged or contaminated food products. The direction and breadth of pest-control strategies in the food industry have changed significantly over the past few years, creating the need to optimize insecticides through improved integrated pest management (IPM) techniques, specifically through the identification of new control agents that are low in mammalian toxicity, as well as any factors that might affect susceptibility to these agents. There is currently renewed interest in developing reduced-risk, low toxicity chemicals that can be effectively utilized in a setting in which grain and other food commodities are vulnerable to insect infestation, as a means of replacing outdated, and at times, less effective methods of insect control. Over the past decade, developed countries have made significant progress toward alternative insect control strategies by employing a variety of applied insecticides. Two classes of insecticide include natural pyrethrum and insect growth regulators (IGRs), which are substances that mimic insect hormones essential to normal development and reproduction. Pyrethrin is a highly efficient, broad spectrum, botanical insecticide that causes a rapid knockdown in exposed insects. Synergists are used to extend the economic usage of natural pyrethrins and because pyrethrum is rapidly metabolized, it is often mixed with a synergist. Methoprene, a juvenile hormone analog, is labeled as an aerosol and surface treatment inside mills, warehouses and other food storage facilities. There is little recent research with large-scale aerosol applications in stored-food facilities; furthermore, there are few published references regarding the efficacy of using methoprene in combination with synergized pyrethrin, in aerosol form. Therefore, the purpose of this research was to evaluate the use of aerosol applications of two aerosol concentrations on flour and finished stored-product packaging materials for the control of Tribolium spp. Results of this research show that T. castaneum are effectively controlled with 1% aerosol application, while the 3% formulation is required to effectively control T. confusum. With regards to the various packaging material surfaces, few differences between the surfaces emerged.