No description available.

Doctor of Philosophy / Department of Entomology / Franklin Arthur / James R. Nechols / The Indianmeal moth, Plodia interpunctella Hubner, is capable of infesting a number of different commodities including a wide variety of grains, nuts and finished stored products. Therefore, control of the Indianmeal moth is especially needed in areas where food is being stored for human consumption. Increased concerns of consumers and producers regarding the impact of conventional insecticides on the environment and on human health has prompted scientists and the agricultural chemical industry to search for insecticides that do not affect mammalian nervous systems and have limited effects on non-target organisms. One group of insecticides with reduced risks is insect growth regulators (IGRs), which are substances that mimic insect hormones essential to normal development and reproduction. Currently methoprene, a juvenile hormone analog, is labeled for direct application to stored grains, as well as a contact insecticide and as an aerosol application inside mills, warehouses, and indoor food storage facilities. Surface treatments and aerosol space applications can be effective ways to treat the interior surfaces and storage areas of warehouses and food processing facilities. There is little recent research with large-scale aerosol applications in storage sites; furthermore, there are no published references in the scientific literature regarding efficacy of using methoprene alone in aerosol form. Therefore, the purpose of this research was to evaluate the use of surface and aerosol applications of methoprene on finished stored-product packaging materials and facilities for the control of P. interpunctella. Results of this research showed that while methoprene has good residual activity, and efficacy is unaffected by temperature, surface applications of methoprene on packaging materials is not as effective for control of P. interpunctella as aerosol applications of methoprene. Aerosol methoprene is highly effective alone and in combination with conventional chemicals for control of eggs and wandering-phase larvae. Simulations with a population growth model make it possible to estimate impact of insecticide treatments at different temperatures and application times on populations of P. interpunctella. Aerosol treatments are also economically viable as part of an overall integrated pest management program.

Master of Science / Department of Grain Science and Industry / Subramanyam Bhadriraju / Spinosad, methoprene, deltamethrin, and deltamethrin plus methoprene, are approved in the United States for treating popcorn. The Indian meal moth, Plodia interpunctella (Hübner), and maize weevil, Sitophilus zeamais (Motschulsky) are two stored-product insects found in popcorn. The efficacy of spinosad and methoprene against P. interpunctella in popcorn were determined in laboratory and field studies. In the laboratory study, eggs (to represent first instars), third, and fifth instars of the laboratory strain of P. interpunctella were exposed to 0.7, 1.4, 2.8 ppm methoprene and 1 ppm spinosad treated popcorn, respectively, to assess larval or adult emergence. In the field study, untreated and treated popcorn samples were placed in vinyl mesh pouches with two mesh-opening sizes and were buried 5 cm below popcorn surface. Pouches with large mesh-opening were used to monitor natural insect infestation between May to October, 2017. Pouches with small mesh-opening were used to conduct laboratory bioassays to evaluate adult emergence of P. interpunctella from eggs after exposed to treated popcorn. Probe traps, food- and pheromone-baited traps, and sticky traps were used to monitor insects in storage bins and cleaning processing facility. The laboratory study showed that there was no P. interpunctella adults emerged from eggs, third, and fifth instars in methoprene treated popcorn during 6 month storage. However, methoprene did not reduce egg-to-larval survival. Larval and adult emergence in the spinosad treated popcorn was significantly lower than controls. Field study showed that there was no adult emergence in methoprene treated popcorn in most cases, and significantly lower adult emergence in spinosad treated popcorn compare to control. P. interpunctella larva was the major insect found in large pouches. The red flour beetle, Tribolium castaneum (Herbst), and P. interpunctella were primary insect species captured by probe traps, food- and pheromone-baited traps and sticky traps over the six months’ study. These results suggested that methoprene could reduce P. interpunctella adult emergence. Spinosad also effectively suppressed the infestation of P. interpunctella. The field strain of S. zeamais was exposed to spinosad (1 ppm), methoprene (0.7, 1.4, 2.8 ppm), deltamethrin (0.5, 1.0 ppm), and deltamethrin plus methoprene (0.5+1.25, 1.0+2.5 ppm) treated popcorn, respectively, for 1 to 336 h exposure time. Mortality was assessed at 0, 7, 14, and 21 d after transferring to clean popcorn except for methoprene treatments which only counted mortality at 0 d. Progeny and adult emergence were counted after 42 d incubation in clean or original popcorn, respectively. All insecticides showed no delay toxicity against S. zeamais adults. Spinosad caused 100% mortality of S. zeamais after 336-h exposure. Complete progeny reduction and highest adult emergence reduction at 168 h exposure. The highest mortality of S. zeamais was 67.1 and 70.5% in deltamethrin and deltamethrin plus methoprene treated popcorn, respectively. Methoprene showed limited efficacy against S. zeamais including low mortality, progeny reduction and high adult emergence. These results indicated that spinosad was effective against S. zeamais. It is necessary to combine other insecticides with deltamethrin and methoprene to control field strain of S. zeamais in stored popcorn.

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Agradecemos a compreensão. on 2016-03-04T13:53:10Z (GMT) / Submitted by DIEGO FARIA COLA null (coladf@gmail.com) on 2016-03-04T14:12:33Z No. of bitstreams: 1 dissertacao.pdf: 1500361 bytes, checksum: bff7d0721d385321109c2e58dfa5697a (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-03-04T14:43:05Z (GMT) No. of bitstreams: 1 cola_df_me_soro.pdf: 1500361 bytes, checksum: bff7d0721d385321109c2e58dfa5697a (MD5) / Made available in DSpace on 2016-03-04T14:43:05Z (GMT). No. of bitstreams: 1 cola_df_me_soro.pdf: 1500361 bytes, checksum: bff7d0721d385321109c2e58dfa5697a (MD5) Previous issue date: 2016-02-26 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O estudo de características e aplicações de estruturas em escala nanométrica passou a ter um grande interesse de pesquisadores das áreas médicas (humana e animal), ambiental e agrícola. Entre essas estruturas, destacam-se os nanocarreadores, que permitiram a melhoraria da biodisponibilidade de muitos compostos bioativos e a diminuição de possíveis efeitos toxicológicos. Além disso, evitar a ocorrência de altos prejuízos na agropecuária com o desenvolvimento de novos nanocarreadores voltados ao combate de endo e ectoparasitas causadores de grandes prejuízos aos pecuaristas. Fármacos como a Ivermectina e o Metopreno são utilizados no combate de ectoparasitas, no entanto, estes compostos possuem problemas relacionados à biodisponibilidade. Este trabalho teve como objetivo preparar e caracterizar sistemas carreadores lipídicos, como as Nanopartículas Lipídicas Sólidas e os Carreadores Lipídicos Nanoestruturados a fim de melhorar e produzir alternativas para o uso destes compostos visando aplicações em veterinária. Na primeira etapa do trabalho foram preparadas e caracterizadas as nanopartículas lipídicas sólidas e os carreadores lipídicos nanoestruturados contendo os fármacos ivermectina e metopreno. As propriedades físico-químicas das suspensões coloidais, como, diâmetro, polidispersão, potencial zeta, pH e eficiência de encapsulação foram avaliadas em função do tempo. A eficiência de encapsulação alcançada foi acima de 99% para ambos os fármacos, sendo que as suspensões coloidais apresentaram estabilidade durante o período investigado. Os resultados do ensaio de citotoxicidade e genotoxicidade demonstram que as nanopartículas testadas para a encapsulação possibilitaram um aumento na viabilidade celular das células testadas, onde as nanopartículas lipídicas sólidas apresentaram menor toxicidade do que os carreadores lipídicos nanoestruturados. As nanopartículas lipídicas sólidas apresentaram tamanho médio de 285,6 ± 15,8 nm e os carreadores lipídicos nanoestruturados de 257,5 ± 7,4 nm. Todas os carreadores apresentaram morfologia esférica, sem formação de agregados e sem alterações morfológicas após a encapsulação. Portanto, os resultados deste trabalho demostraram que as nanopartículas apresentaram boas características coloidais e a presença ou ausência de drogas não provocam alterações nas características das nanopartículas, possibilitando o desenvolvimento de sistemas carreadores para estas drogas visando aplicações veterinárias. / The study characteristics and structures of applications in nanoscale have a great interest of researchers in medical areas (human and animal), environment and agriculture. Among these, the nanocarriers stand out, that allowed improve the bioavailability of many bioactive compounds and the reduction of potential toxicological effects, and prevent the occurrence of high losses in agriculture with the development of new nanocarriers aimed at fighting and endo ectoparasites causing great harm to farmers. Drugs such as Ivermectin and Methoprene are used to combat ectoparasites, however, these compounds possess bioavailability problems related. This study aimed to prepare and characterize lipid carrier systems, such as Solid Lipid Nanoparticles and Nanostructured Lipid carriers to improve and produce alternatives to the use of these compounds targeting applications in veterinary. In the first stage of the work we have been prepared and characterized the solid lipid nanoparticles and nanostructured lipid carriers containing the drug ivermectin or methoprene. The physico-chemical properties of colloidal suspensions such as, diameter, polydispersity, Zeta potential, pH and encapsulation efficiency were measured as a function of time. The encapsulation efficiency was achieved above 99% for both drugs, and colloidal suspensions were stable during the period investigated. The results of the cytotoxicity and genotoxicity test demonstrate that the tested nanoparticles for encapsulating allowed an increase in cell viability of test cells, where the solid lipid nanoparticles had lower toxicity than the nanostructured lipid carriers. The solid lipid nanoparticles have an average size of 285,6 ± 15,8 nm and nanostructured lipid carriers 257,5 ± 7,4 nm. All the carriers had spherical morphology without forming aggregates and without morphological changes after encapsulation. Therefore, the results of this study demonstrated that the colloidal nanoparticles showed good characteristics and the presence or absence of drug do not cause changes in the characteristics of the nanoparticles, allowing the development of carrier systems for drug targeting these veterinary applications. / FAPESP: 2013/24788-6

Juvenile hormone (JH) is one of the principal hormones that regulate insect development and reproduction. Accumulating evidence suggests that Methoprene-tolerant (Met) protein is a nuclear receptor of JH. Work by others has shown that Met is capable of binding JH at physiological concentration. An RNAi knockdown of Met causes down-regulated expression of JH-responsive genes and a phenotype similar to that observed in JH-deficient insects, suggesting that Met is required for mediating physiological and molecular responses to JH. The work in this report aims to understand the mechanisms underlying gene regulation by JH via Met. Met is a bHLH-PAS (basic-helix-loop-helix Per-ARNT-Sim) family protein. Many proteins in this family function as heterodimers formed with other proteins of this family. In a yeast two-hybrid screening, we discovered that another bHLH-PAS family protein, FISC, interacts with Met in the presence of JH. FISC is also required for JH functions as an RNAi knockdown of FISC down-regulated JH-responsive genes. To elucidate how Met and FISC mediate JH functions in gene regulation, we employed molecular biology techniques and characterized the formation of a JH-Met-FISC complex and its actions in activating gene expression using mosquito Aedes aegypti as a model. My results demonstrated that Met and FISC forms a complex when JH is present via their conserved N-terminal domains. The complex then binds to E box-like sequences presented in the promoter of JH-responsive genes to activate gene expression. This mechanism also applies to the fruit fly Drosophila melanogaster, suggesting that it is a conserved action of JH in insects. Further studies showed that DNA-binding by Met and FISC requires the basic regions of the bHLH domains of both proteins. Lastly we identified a consensus binding-site of Met and FISC. / Ph. D.

No description available.

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.

The evolution of nonreproductive castes is a defining characteristic of eusociality. The function and developmental regulation of the altruistic worker and soldier caste is the central element contributing to major advantages of eusociality over solitary animals. The soldier caste is the first evolved sterile caste in termites. Their primary function is believed to be colony defense. However, the function and development of termite soldiers remains largely unknown. Because of their apparent morphological adaptation for fighting and their limited behavior repertoire, our understanding of colony defense by termite soldiers is limited to their physical defense. In addition, we know little about the molecular mechanisms mediating soldier development. In Chapters 2 and 3 I discuss the role of the soldier caste under competition risk. By exposing the Eastern subterranean termite Reticulitermes flavipes to cues of a competitor termite species, I found that exposure to competitor cues reduced feeding, compromised growth and survival of R. flavipes workers. The presence of R. flavipes soldiers largely ameliorated these negative impacts. At the transcriptional level, R. flavipes soldiers can counteract the effects of competitor cues on worker head gene expression. This counteracting effect seems to be associated with genes in metabolism and immunity. These studies demonstrate that competition can affect a termite colony’s fitness by either competitors physically invading the colony and causing damage or cues from competitors inducing a stress response in termite colony members. More importantly, soldiers can contribute to colony fitness by physically engaging in combat, but also by enhancing colony members’ survival under competitor-cue exposure. In Chapter 4, I describe the molecular mechanism mediating soldier-caste differentiation. I cloned the full length cDNA sequence of the R. flavipes Methoprene-tolerance (Met) gene, a gene encoding a putative receptor for juvenile hormones. Using RNA interference, I studied the function of Met and found that this gene essentially mediates the JH-dependent soldier-caste differentiation in termites.

Doctor of Philosophy / Department of Entomology / James F. Campbell / Kun Yan Zhu / Aerosol applications of reduced risk insecticides such as synergized pyrethrin and insect growth regulators (IGR) are part of food industry integrated pest management programs. Since aerosols cannot penetrate into hidden areas exploited by pests such as the red flour beetle, Tribolium castaneum, the potential for these insecticides to effect beetle populations was evaluated. Because IGRs do not cause immediate mortality, the potential of horizontal transfer for an IGR from treated to untreated individuals was also examined. Results showed that when untreated T. castaneum, larvae or pupae, were added to flour containing methoprene, IGR, treated larvae, pupae or adults, the untreated individuals exhibited evidence of methoprene exposure (external deformities and reduced survival). Evaluation of the different mechanisms of transfer indicated that contact with methoprene treated individuals or flour that had been in contact with treated individuals may be the primary method of methoprene transfer. Since aerosols are often applied as a combination of IGR and pyrethrin with a carrier, the effect of these components was evaluated. Applications of synergized pyrethrin caused knockdown of adults but affected adults recovered and progeny production was not effected. Exposure of eggs to these insecticides reduced egg hatch. Food material accumulations inside food facilities can potentially increase or reduce insecticide efficacy. Evaluation of different flour residue levels, representing different sanitation levels, revealed that sanitation alone reduced immature development. As flour residue depths increased more individuals developed into adults but very few developed in the insecticide treatments. Food facilities that use aerosol insecticides apply them at regular intervals, so the cumulative effects of these treatments were considered. Experiments evaluating repeated insecticide exposures indicated that the direct morality from synergized pyrethrin not the horizontal transfer of methoprene was the primary factor in population reduction. Overall findings suggested that methoprene is highly mobile between different surfaces. Exposure of untreated individual beetle larvae to treated larvae or pupae or to flour that has been in contact with exposed beetles can have detrimental effects on development or survival, but these effects may be highly variable and even in cumulative exposures the overall level of population suppression is limited.

Doctor of Philosophy / Department of Grain Science and Industry / Subramanyam Bhadriraju / Hulya Dogan / Larvae of the Indian meal moth, Plodia interpunctella (Hübner), have the ability to invade or penetrate packaging materials and infest the food within. Energy bars with three polypropylene packaging types were challenged with eggs (for first instars), third, and fifth instars of P. interpunctella to determine package resistance to larvae at 28°C and 65% r.h. to determine which provided the greatest protection against P. interpunctella larval penetration. Third and fifth instars showed a higher propensity to infest all packaging variations. First instars showed a reduction in package pentration abiltiy compared to third and fifth instars. Methoprene is an insect growth regulator labeled in the USA for use as an aerosol spray, a residual surface treatment, and as a grain protectant, and recently has been impregnated into a polymer-based packing film to prevent insect infestations of packaged products. The objectives of these studies were 1) determine the effect of short term exposure time and temperature on four week old larvae, continual exposure on egg-to-adult emergence of beetles, and sub-lethal effect on adults of the red flour beetle, Tribolium castaneum (Herbst) and warehouse beetle, Trogoderma variabile Ballion, on the inside and outside surfaces of methoprene-treated woven packaging material at 27 and 32°C at 60% r.h.; 2) evaluate fecundity, egg hatch, and egg-to-adult emergence of T. castaneum and T. variabile, when exposed to two methoprene-impregnated polymer packaging on the inside and outside surfaces at 27 and 32°C at 60% r.h.; and 3) determine the effect of methoprene-treated foil packaging on larval emergence, penetration, and invasion ability of T. variabile and P. interpunctella at 27°C and 60% r.h. Short term exposure results indicated that adult emergence from larvae of T. castaneum and T. variabile decreased with increasing exposure time. The number of eggs laid per female of T. castaneum and T. variabile did not vary from their controls. Continual exposure demonstrated 100% supression of T. castaneum adult emergence, irrespective of exposure to outside or inside surfaces. T. variabile exposed to inside surfaces were unaffected and normal adult emergence was reduced in those exposed to outside surfaces.. The number of T. variabile eggs laid per female was not significantly different among polymer packaging types. The methoprene-treated polyethylene terephthalate to polyethylene packaging, PET-PE reduced the number of T. castaneum eggs laid per female. Both polymer packaging reduced the percent hatch of both species. No T. castaneum adults emerged on the inside surface of PET-PE and both sides of the polyethlyene to polyethylene (PE-PE). Egg-to-adult emergence of T. variabile was arrested at the pupal stage on the outside surface of PE-PE packaging. The PET-PE packaging greatly reduced the number of normal adults by 87 to 97% when exposed to inside surfaces at both temperatures. The foil packaging had no significant effect on hatch of either species. T. variabile were unable to penetrate/invade any foil packages. P. interpunctella invaded all packaging containing pinholes. Therefore, continual exposure of T. castaneum and T. variabile to methoprene impregnated packaging could be a vaiable tool to protect food packages.