Thesis (Ph. D.)--University of Missouri-Columbia, 2003. / Typescript. Vita. Includes bibliographical references (leaves 78-86). Also available on the Internet.

Efficacy of Bacillus thuringiensis Berliner against the first three larval instars of Heliothis virescens (F.) was studied under greenhouse and field conditions. The primary objective of these studies was to determine the susceptibility of the tobacco budworm to Dipel, a commercial preparation of Bacillus thuringiensis. Both greenhouse and field studies showed that first-instar larvae were more susceptible to all dosage levels of Dipel tested (0.125, 0.25, 0.5, and 1.0 1b/A). The higher rates of 0.5 and 1.0 1b/A gave better control and remained effective for longer periods against the first-, second- and third-larval instars than did the lower rates of 0.125 and 0.25 1b/A. The efficacy of all Dipel rates decreased significantly with each subsequent post-application-date bioassay, and only the higher dosages of 0.5 and 1.0 1b/A remained effective at the three-day residue period in the field studies. The addition of a feeding stimulant, Coax, at 0.25 1b/A, to all Dipel treatments resulted in a significant increase in larval mortality of all instars tested. Dipel plus Coax exhibited longer residual activity than did Dipel alone, indicating that Coax may protect the spores and toxin from degradation by the direct sunlight. When second-instar tobacco budworm larvae were allowed to feed on Dipel-treated terminals for 6, 18 or 30 hours and then transferred to untreated diet, they showed a great capacity to recover from the effect of the toxin. However, as the Dipel rate or exposure time was increased, the ability of larvae to recover was decreased. Dipel did have an effect on the biological activity of treatment-surviving larvae. Larval and pupal development was delayed, pupal weight was decreased, and female fecundity was reduced. In all cases the changes were proportional with the Dipel rate and with the length of exposure. No apparent effect was shown relative to adult longevity or egg viability for those surviving Dipel treatments as larvae. When Dipel was applied directly on tobacco budworm eggs, it had no effect on egg hatch but did significantly affect survival of newly-emerged larvae, indicating the larvae consumed a lethal dose of the toxin during eclosion. The compatibility of Dipel in combination with Geocoris punctipes (Say) against first-instar larvae of Heliothis virescens was also studied under greenhouse and field conditions. Results of these studies indicated the possibility of utilizing the two biological agents to suppress tobacco budworm populations in the field. A minimum density of one Geocoris nymph per 4 row feet is required, plus an effective field rate of Dipel. When adult tobacco budworms were fed on Dipel at 32000 IU/ml, their longevity and fecundity were significantly reduced. An adult feeding stimulant is needed to lure the adults to a Dipel source.

Thesis (Ph. D.)--University of Missouri-Columbia, 1998. / Typescript. Vita. Includes bibliographical references (leaves [224]-254). Also available on the Internet.

Thesis (Ph. D.)--University of Missouri-Columbia, 1998. / Typescript. Vita. Includes bibliographical references (leaves [224]-254). Also available on the Internet.

Submitted by (edna.saturno@ufrpe.br) on 2016-11-24T15:08:38Z No. of bitstreams: 1 Gilberto Santos Andrade.pdf: 539005 bytes, checksum: 575c35c9686dfc82cb2bf5b3882e07f1 (MD5) / Made available in DSpace on 2016-11-24T15:08:38Z (GMT). No. of bitstreams: 1 Gilberto Santos Andrade.pdf: 539005 bytes, checksum: 575c35c9686dfc82cb2bf5b3882e07f1 (MD5) Previous issue date: 2007-02-01 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / One important aspect of maximizing pest natural mortality factors is to enhance the action of natural enemies into integrated pest management programs. Species of the egg parasitoid Trichogramma have been investigated and used successfully around the world. To boost the utilization of these parasitoids as biological control agent, basic studies on species selection regarding the interactions with environmental conditions and host plants is needed. In Brazil, Trichogramma pretiosum Riley is recommended to control lepidopteran pests of cotton. Thus, this research evaluated the potential of other three Trichogramma species in comparison to T. pretiosum parasitizing eggs of tobacco budworm Heliothis virescens (Fabr.) under constant (20, 25 and 30oC) and alternated (immature development at 25oC and adult submitted to 20 and 30oC) temperatures. Additionally, the best performing species at these temperature regimes were tested when offering egg of the host tobacco budworm on cotton leaves of two varieties (ca. colored BRS Safira and white fibers BRS 8H). The species studied were: Trichogramma exiguum Pinto & Platner, Trichogramma atopovirilia Oatman & Platner, Trichogramma acacioi Brun, Moraes & Soares and T. pretiosum. The evaluated parameters were: parasitism rate and its viability, sex ratio of the progeny and number of descendents emerged per egg parasitized. Overall, T. atopoviriliaexhibited the best performance through all temperature regimes studied. T. pretiosum and T.exiguum achieved similar parasitism rate of H. virescens eggs, while T. acacioi showed the lower performance. Based on the results above, parasitism of eggs offered on cotton leaves was conducted with the species T. exiguum, T. pretiosum and T. atopovirilia. The cotton variety affected the parasitism rate of T. exiguum and T. atopovirilia resulting in lower parasitism when eggs were offered on BRS Safira cotton leaves, but it was similar for T. pretiosum. Therefore, under constant and alternating temperatures and host exposure on cotton leaves all species parasitized eggs of H. virescens, but T. atopovirilia exhibited better performance in comparison to the others. Therefore, further studies should be conducted looking for other field aspects related with the use of this parasitoid such as dispersion and field parasitism. / Espécies do gênero Trichogramma têm sido pesquisadas em todo o mundo para o controle biológico de lepidópteros-praga. Para que a eficiência desses parasitóides seja aumentada faz-se necessário a realização de estudos básicos visando à seleção de espécies em relação ao ambiente e seus hospedeiros. No Brasil, a espécie Trichogramma pretiosum Riley é a mais utilizada para o controle de lepidópteros-praga do algodoeiro. Assim, comparou-se o desempenho de outras espécies tais como: Trichogramma exiguum Pinto & Platner, Trichogramma atopovirilia Oatman & Platner e Trichogramma acacioi Brun, Moraes & Soares (Hym.: Trichogrammatidae) parasitando ovos de Heliothis virencens (Fabr.) (Lep.: Noctuidae) sob temperaturas constantes (20, 25 e 30oC) e alternadas (fase imatura a 25oC e adultos submetidos a 20 e 30oC para parasitismo). Adicionalmente, as espécies com melhor desempenho nas temperaturas estudadas foram testadas oferecendo ovos de H. virescens em folhas de algodoeiro das cultivares BRS 8H e BRS Safira. As variáveis avaliadas foram: porcentagem de parasitismo e viabilidade; razão sexual e número de indivíduos por ovo. T. atopovirilia foi a espécie com melhor desempenho em todas as condições térmicas estudadas constatado através da maior porcentagem de parasitismo. T. pretiosum e T. exiguum apresentaram a mesma taxa de parasitismo em ovos de H. virescens e T. acacioi obteve a menor taxa de parasitismo. Com base nesses resultados, o estudo oferecendoovos em folhas das duas cultivares de algodoeiro foi conduzido com T. exiguum, T. pretiosum e T. atopovirilia. As cultivares de algodoeiro afetaram o parasitismo de ovos de H. virescens de T. atopovirilia e T. exiguum, sendo inferior quando os ovos foram oferecidos em folhas de BRS Safira. Não houve efeito das cultivares no parasitismo de H. virescens por T. pretiosum. Os resultados evidenciam que T. atopovirilia e T. exiguum apresentaram melhor parasitismo de ovos depositados em BRS 8H e o parasitismo de T. atopovirilia foi superior aquele de T. pretiosum quando ovos encontravam-se sobre a cultivar BRS Safira. Assim, futuros estudos devem ser conduzidos com T. atopovirilia parasitando ovos de H. virescens em condição de campo e onde sejam analisadas variáveis como quantidade de parasitóides a serem liberados para a obtenção de taxas de parasitismo satisfatórias.

In this study, a sensitive high performance thin-layer chromatography method (HPTLC) was developed to detect lipids in microgram (ug) quantities of hemolymph. The effect of age, diurnal rhythms, and flight activity on hemolymph 1,2-diacylglyceride concentrations in male and female Heliothis zea (Boddie) (Lepidoptera: Noctuidae) were quantitated. The HPTLC method involved a two-stage, one dimensional solvent system, 1 ). benzene:ether(diethyl):ethanol:acetic acid, 50:40:2.0:0.2; 2). hexane:heptane: ether (diethyl): acetic acid, 63:18:18:1. The solvent system separated alkanes, steryl ester, mono-, di- and triacylglycerides, free fatty acids and cholesterol. Lipids were detected with a fluorescent reagent, 1 mM 6-p-toludino-2-napthalenesulfonic acid (TNS) in 95% methanol. Detection sensitivities were 50 ng qualitatively and 500 ng quantitatively. Most male and female Heliothis zea flight activity occurred between 17⁰⁰ and 3⁰⁰ EST each day. Male flight activity after the day of eclosion through 6 days was 0.8, 1.0, 7.0, 3.3, 3.8, 2.8 and 4.0 times the flight activity of the females. Females had a unimodal pattern of flight activity between 17⁰⁰ and 19⁰⁰ Males had a bimodal pattern between 17⁰⁰ and 19⁰⁰, but another higher peak of flight activity existed between 20⁰⁰ and 24⁰⁰. The hemolymph of Heliothis zea males and females prior to flight (17⁰⁰) was composed of mainly diacylglycerides (50% w/w), and triacylglycerides (35% w/w). In addition, cholesterol esters (2%) and less than 1% monoacylglycerides and cholesterol were present, but free fatty acids were not detectable ( <0.5 ug/ul). The lipid class, 1,2-diacylglycerides, apparently are used for flight energy. They occured in the highest concentrations in both males and females prior to flight (17⁰⁰) (approximately 32 ug/ul), then decreased steadily throughout flight to approximately 16 ug/ul at 2⁰⁰ when flight activity was reduced or ceased. Males and females appear to have similar 1,2-diacylglyceride concentrations prior to, during and after flight activity. / M.S.

A strategy for the discovery of new insect growth regulators from arid lands plants was developed. Plant genera with a history of toxicity, medicinal use, or incorporation in native american cultures were selected. Forty-five species from twenty-one families were collected, extracted, and tested for biological activity on the large milkweed bug, Oncopeltus fasciatus. Eight extracts were toxic and the extract of Nama hispidum (Hydrophyllaceae) caused nymphs to undergo precocious metamorphosis to an adult. The active component of N. hispidum was precocene II (6,7-dimethoxy-2,2-dimethyl chromene). Nine Nama species, representing four of the five sections in the genus were gathered from Hawaii, California, Nevada, Arizona, New Mexico, and the Dominican Republic. In addition to the insect anti-hormone, precocene II (PII), present in N. hispidum, N. rothrockii contained at least two different insect juvenile hormone mimics, and N. sandwicense contained insect anti-hormone and insect juvenile hormone mimics. Fifth instar larvae of Heliothis zea were used as model insects to distinguish between post-ingestive intoxication and feeding deterrency during exposure to PII. Larvae fed artificial diet with PII were deterred from eating, and had retarded weight gain, growth, and development. Insects grown on diet with PII consumed less food, could not digest the food, or convert ingested food to body mass. in vivo radiotracer studies with the nutrient, ¹⁴C linoleic acid, show a lower rate of transport and incorporation of radioactivity into fat body tissue. A change in the midgut epithelial cells from PII resulted in reduced transport efficiency and retarded larval growth and development. Scanning electron microscopy of the midgut epithelia indicated that cytotoxic damage is induced by PII. Observed changes in the midgut epithelial cells are consistent with a destructive alkylation of cell structures by PII.

No description available.

Microplitis croceipes is a solitary endoparasitic wasp that oviposits in the hemocoel of Heleothis virescens larvae. Upon parasitization, the host larvaes physiology is altered; resulting in a compromised immune system and a decrease in the production of some vital proteins resulting in a terminal post-wandering prepupal state. Teratocytes, cells derived from the extraembryonic serosa of the parasitic wasp, mimic symptoms of parasitization when injected into host larvae, independent of other factors like polydnavirus and venom. Some of the inhibition of protein synthesis can be attributed to proteins secreted by the teratocytes (teratocyte secretory proteins or TSP). A fraction of TSP between 330 kDa inhibits protein synthesis in vivo, in the in vitro fat body and testes assays, and in the rabbit reticulocyte lysate and wheat germ extract assays. This fraction, however, has no effect on nucleic acid synthesis. Its effect on protein synthesis is dose dependent and exposure time sensitive. A 13.9 kDa protein isolated from TSP and expressed in a baculovirus system seems primarily responsible for the inhibition. Although TSP14 production was low, it did bind to the cell surface, enter the cell, and inhibit protein synthesis as the 330 kDa factor did.

The crystal (Cry) toxins from Bacillus thuringiensis (Bt) display high specificity and toxicity against relevant insect pests and the use of Bt-based products continues to contribute to insect pest management. To protect this investment, further its potential, and investigate possible unintended effects, various research questions have been proposed. One issue related to Bt usage is the evolution of pest resistance to Bt toxins. The midgut epithelium is targeted by Cry toxins killing enterocytes, facilitating invasion of the hemocoel, leading to septicemia and mortality. While resistance may emerge from alterations to these steps, most research efforts have been focused on reduced toxin binding to midgut receptors as resistance mechanism. Lepidopteran crop pest Heliothis virescens strains have been hypothesized to have enhanced midgut proliferation and differentiation of stem cell populations allowing for regeneration and resistance to diverse Cry toxins. However, the molecular mechanisms involved are not known. We developed a flow cytometry method to monitor stem cell proliferation and differentiation to compare midgut regenerative responses to Cry intoxication in larvae from susceptible and Bt-resistant strains of H. virescens. The structure of the epithelial healing response was studied in vivo using hematoxylin-eosin stained midguts derived from larvae fed Cry1Ac toxin. We detected less regenerative cells in midguts from a Bt-susceptible strain (YDK) compared to midguts from resistant (KCB and CXC) strains, and an overall increase in the total number of cells per unit surface area in KCB midguts. Using primary midgut cell cultures, the midgut regeneration response to Cry1Ac in CXC was an increase in available differentiated cells compared to YDK. In contrast, KCB exhibited an increased abundance of stem cells compared to both YDK and CXC. Using a differential proteomics approach we characterized the proteins secreted by H. virescens midgut cells in response to Cry1Ac and identified a relevant role for arylphorin in promoting midgut regeneration in response to Cry1Ac and DiPel intoxication in both susceptible and resistant H. virescens larvae. The potential fitness costs associated with altered hexamerin transcript expression were monitored using larval bioassays.