Thesis (Ph. D.) in Biological Sciences--University of Maine, 2001. / Includes vita. Advisory Committee: Eleanor Groden, Assoc. Prof. of Entomology, Advisor; Francis Drummond, Prof. of Insect Ecology; Seanna Annis, Asst. Prof. of Mycology; David Lambert, Assoc. Prof. of Plant Pathology; John D. Vandenberg, Research Entomology, USDA-ARS. Includes bibliographical references (leaves 158-175).

Thesis (M.S.)--University of Missouri-Columbia, 2008. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on July 9, 2009) Includes bibliographical references.

Thesis (M.Sc.)--Memorial University of Newfoundland, 2003. / Bibliography: leaves 75-86.

Evaluations of tactics for detection and management of raspberry crown borer, RCB, Pennisettia marginata (Harris) on blackberries were performed in Mississippi from January 2005 - July 2006. Randomized split plots with infested blackberries and certified nursery stock of two recommended cultivars, thorny ?Chickasaw? and thornless ?Apache? were planted. Pest management tactics targeting RCB included drenches: chemical insecticide bifenthrin, recently registered for RCB management; experimental pesticide E2Y45; and entomopathogenic nematodes Steinernema feltiae. Insect pheromones (E,Z)3-13-octadecadien-1-ol and (E,Z)3-13-octadecadien-1-yl-acetate were evaluated as RCB lures. Effects of fungicides captan, pyraclostrobin + boscalid, lime-sulfur and insecticide acetamiprid on viability and infectivity of entomopathogenic nematode species, S. feltiae and Heterorhabditis bacteriophora were evaluated in laboratory bioassays with host greater wax moth larvae. RCB larvae and soil pests on and around infested blackberry plants were observed. Bait traps with 5 mg of (E.Z)3-13-octadecadien-1-ol captured an adult RCB on October 19, 2005. Laboratory bioassays indicated that lime-sulfur and pyraclostrobin + boscalid decrease nematode movement and infectivity.

Entomopathogenic fungi are highly susceptible to the damaging effects of solar radiation. Attempts to protect entomopathogenic fungi from solar radiation have been, for the most part, unsuccessful. A new strategy for formulating entomopathogenic fungi for protection from solar radiation and desiccation has been developed tested using the acridid entomopathogen Metarhizium anisopliae var acridum (IMI 330189). This strategy involves coating spores with water-soluble materials that provide protection from solar radiation and enhance spore survival during drying. Development of this formulation involved the following: 1) production of an infective spore-type in liquid culture that could survive drying; 2) coating spores during an air-drying process; 3) reducing formulation particle size for oil suspension; 4) testing the effects spore coating on spore-tolerance simulated sunlight; and 6) testing the effects of spore coating infectivity to Schistocerca americana. Aerial conidia, submerged conidia, and blastospores produced in a high-osmolality liquid medium all had high desiccation tolerance relative to blastospores produced in Adamek's media. Blastospores produced in high osmolality medium were the most infective to S. americana in an aqueous 20% molasses solution followed by submerged conidia and aerial conidia, with LT50 values (95% C.I.) at 1 x 106 spores/insect of 7.8 d (6.7 to 9.0 d), 10.5 d (9.5 to 11.6 d), 14.6 d (11.9 to 18.0 d), respectively. Comparisons were made among cell-wall characteristics of these spore-types, including cell-wall thickness, lectin-binding, charge, and hydrophobicity. An optimal spore-coating formulation was selected on the basis of spore survival and germination over time after air-drying, particle-size reduction, and storage at 28 Ë C. This spore-coating formulation, consisting of skim milk, Kraft lignin (Curan 10®) and glycerol, greatly improved the tolerance of aerial conidia and submerged conidia to simulated sunlight, increasing the LT50 (95% C.I.) of aerial conidia from 4.0 hr (3.1-5.1) to 17.0 hr (12.5-23.0). The spore coating formulation decreased the infectivity of spores in oil to adult S. americana; reducing the LT50 values of aerial conidia at a dose of 1 x 105 spores / insect from 5.8 d (4.9-6.9 d) to 8.2 d (7.3-9.3 d). / Ph. D.

Photorhabdus are motile Gram-negative bacteria that have a mutualistic association with entomopathogenic Heterorhabditis nematodes. Nematodes vector the bacteria from one insect host to another, while the bacterial symbiont produces toxins and secondary metabolites that kill that the insect host. In this study, we characterize the bacterial symbiont of Heterorhabditis sonorensis, recently discovered in the Sonoran desert. Biochemical and molecular methods including sequence data from five genes: 16s rDNA, gyrB, recA, gltX, dnaN were considered. Evolutionary relationships of this new Photorhabdus subsp. were inferred considering maximum parsimony and Bayesian analyses. We also surveyed for secondary metabolites (SM) produced by this microorganism, considering HPLC and mass spectrometry analyses. SM crude extracts showed activity against the corn ear worm Helicoverpa zea, the root-knot nematode (Meloidogyne incognita), the bacterium Pseudomonas syringae, and the fungus Fusarium oxysporum; and were more toxic that those produced by related species. Results from these studies showed that Photorhabdus l. sonorensis' secondary metabolites have potent antagonistic activity against these plant pathogens.

Os fungos Batkoa sp. e Furia sp.(Entomophthorales), embora ocasionalmente ocorrendo de forma epizoótica em populações das cigarrinhas-das-pastagens e cana-deaçúcar, nunca foram avaliados quanto ao nível de incidência e potencial de controle biológico. Esse estudo teve por objetivo avaliar a ocorrência natural, produção "in vitro" e preservação de micélio seco desses fungos. Nos trabalhos de produção incluiu-se como padrão o fungo Neozygites floridana, representante de outra família de Entomophthorales. O estudo sobre a ocorrência natural foi realizado na região de Pindamonhangaba, SP, no período de 22 de janeiro a 9 de fevereiro de 1998 com o objetivo de avaliar a incidência de Batkoa sp. na população de Mahanarva fimbriolata em pastagem de capim napier e de Furia sp. sobre Deois schach em pastagem de Brachiaria sp. O estudo sobre produção foi realizado em condições de laboratório com o objetivo de desenvolver meios líquidos para a produção de micélio ou corpos hifais de Batkoa sp., Furia sp. e N. floridana, tendo como base o meio líquido para cultura de tecido de inseto Grace’s suplementado, o qual permite bom crescimento do último patógeno. Finalmente, estudou-se o efeito da combinação de dois dessecantes (sílica e glicerol) com dois redutores de oxigênio (vácuo e Agelessâ ZPT-200) na preservação de micélio seco de Batkoa sp. e Furia sp. sob temperaturas de 3°C e 23°C. Concluiu-se que: Batkoa sp. e Furia sp. causam doença na população de M. fimbriolata e D. schach, respectivamente; Furia sp. possui maior potencial epizoótico que Batkoa sp.; Batkoa sp., Furia sp. e N. floridana se assemelham quanto ao crescimento em meios adicionados de sais, vitaminas e aminoácido, e em meio completo; A adição de sais ao meio básico proporciona um aumento significativo na produção das três espécies de fungos, e maior do que a adição de vitaminas e aminoácidos; A mistura dos sais CaCl2.2H2O (0,06%), KCl (0,28%), MgCl2.6H2O (0,16%), MgSO4.7H2O (0,2%), NaHCO3 (0,03%) e NaH2PO4.H2O (0,1%) proporciona maior produção dos fungos do que os sais avaliados isoladamente na concentração de 0,83%; O sulfato de magnésio (MgSO4) pode ser utilizado como único sal, na concentração de 0,83%, visando a produção de Batkoa sp., Furia sp. e N. floridana; Esses fungos se assemelham quanto ao crescimento em meios contendo diferentes fontes de carbono, mas se diferenciam em meios com diferentes fontes de nitrogênio; O meio contendo 2,66% de glucose proporciona uma produção das três espécies de fungos significativamente maior do que o meio com 2,66% de sacarose; A adição de 0,1% de monossacarídeos no meio com 2,66% de sacarose não aumenta significativamente a produção dos fungos; O extrato de levedura proporciona maior produção de Batkoa sp.; A concentração de 0,5% de extrato de levedura é a mais adequada para a produção vegetativa de Batkoa sp.; A combinação de extrato de levedura + extrato de carne + leite desnatado proporciona a maior produção de Furia sp.; A combinação de extrato de levedura + leite desnatado proporciona o segundo maior rendimento de Furia sp., sendo mais adequada para a produção do fungo devido ao menor custo; A concentração de 1% da mistura dessas fontes de nitrogênio é adequada para a produção vegetativa de Furia sp.; A combinação de extrato de levedura + peptona + leite desnatado proporciona a maior produção de N. floridana; A concentração de 3% da mistura dessas fontes de nitrogênio é adequada para a produção vegetativa de N. floridana; A combinação de sílica + Ageless® prolonga a sobrevivência desses dois fungos formulados como micélio seco até 90 dias, armazenados a 3°C e 23°C; O uso de vácuo como redutor de oxigênio não prolonga a sobrevivência desses dois fungos, mesmo em associação com dessecantes; A temperatura de 3°C provoca uma ligeira queda no potencial de crescimento de Batkoa sp., porém prolonga a sobrevivência do fungo até 90 dias, independentemente do tratamento. / The fungi Batkoa sp. and Furia sp. although occasionally occurring at epizootic levels in populations of spittlebug pests of pasture and sugarcane, have never been evaluated as to natural incidence level nor biological control potential. This study was designed to evaluate the natural occurrence, in vitro production and dry mycelium preservation of these fungi. Neozygites floridana, a representative of another family of Entomophthorales, was included as a standard in the production studies. The study on natural occurrence was done in Pindamonhangaba county, São Paulo State, from January 22 to February 9, 1998, to evaluate the incidence of Batkoa sp. in the Mahanarva fimbriolata population on pasture of napier grass, and Furia sp. in Deois schach, on Brachiaria sp. pasture. The study on production was done in laboratory conditions with the purpose to develop liquid media for the production of mycelium or hyphal bodies of Batkoa sp. Furia sp. and N. floridana, using as a base a liquid medium for insect tissue culture [Grace’s supplemented (HyQ® TNM-FH, #SH30280.02)], which allowed good growth of the latter pathogen. Finally, the effect of combinations of two desiccants (silica and glycerol) with two oxygen reducers (vacuum and Ageless® ZPT-200) on the preservation of dry mycelium of Batkoa sp. and Furia sp. under temperatures of 3º C and 23º C was studied. It is concluded that: Batkoa sp. and Furia sp. cause disease in M. fimbriolata and D. schach sp. populations, respectively. Furia sp. has higher epizootic potential than Batkoa sp. Batkoa sp., Furia sp. and N. floridana are similar concerning their growth patterns in media with added salts, vitamins and amino acids, as well as in the complete one. The addition of salts to the basic medium causes a significant increase in biomass production of the three fungal species, and higher than that from the addition of vitamins and amino acids. The mixture of the salts CaCl2.2H2O (0,06%), KCl (0,28%), MgCl2.6H2O (0,16%), MgSO4.7H2O (0,2%), NaHCO3 (0,03%) and NaH2PO4.H2O (0,1%) afforded higher production of the three fungi than the salts solely evaluated, at the concentration of 0,83%. MgSO4 can be used as sole salt for the production of Batkoa sp., Furia sp. and N. floridana. These fungi are similar concerning their growth patterns in media with different sources of carbone, but different in media with different sources of nitrogen. The production of the three fungal species is significantly higher in medium containing 2.66% glucose than in 2.66% sucrose. The addition of 0.1% monossacarides to media containing 2.66% sucrose does not increase significantly biomass production. Yeast extract allows the highest production of Batkoa sp. The concentration of 0.5% yeast extract is the most suitable for vegetative growth of Batkoa sp. The combination of yeast extract + beef extract + skim milk allows the highest production of Furia sp. The combination of yeast extract + skim milk allows the second highest production of Furia sp., and is the most suitable for mass production due to the lower cost. The 1% concentration of the mixture of these nitrogen sources is suitable for vegetative growth of Furia sp. The combination of yeast extract + peptone + skim milk allows the highest production of N. floridana. The 3% concentration of the mixture of these nitrogen sources is suitable for the vegetative growth of N. floridana. The combination of silica + Ageless® extends the survival of these two fungi formulated as dry mycelium for 90 days, stored at 3º C and 23º C. The use of vacuum to reduce oxygen does not extend the survival of these fungi, even in association with desiccants. Storage at 3º C causes a slight decrease in growth potential of Batkoa sp., but it extends fungus survival for 90 days, independent of other treatments.

Esta pesquisa teve o objetivo de avaliar o efeito de derivados das meliáceas Azadirachta indica (nim), Melia azedarach e Trichilia pallida e os fungos entomopatogênicos Metarhizium anisopliae (isolado 1037) e Beauveria bassiana (isolado 634) visando ao controle do cupim subterrâneo Heterotermes tenuis. Os derivados vegetais foram avaliados em relação à toxicidade e repelência para o cupim. A formulação comercial Nimkol®, à base de folhas de nim, foi tóxica para o cupim, em concentrações de 0,3% de ingrediente ativo ou superiores. Não foi constatada a transmissão do Nimkol de indivíduos alimentados com esse produto para outros indivíduos. Os demais extratos vegetais estudados não apresentaram toxicidade significativa para H. tenuis. Nenhum dos produtos avaliados foi repelente a ponto de evitar a construção de galerias em placas com ágar ou provocar o fechamento das mesmas. Entretanto, em testes de escolha em arenas de preferência, H. tenuis demonstrou repelência pelo extrato aquoso de folhas de nim (5%). Os demais derivados avaliados, incluindo o Nimkol, não foram repelentes para o cupim. O efeito dos derivados vegetais e dos entomopatógenos na sobrevivência de H. tenuis, em forma independente e associada, foi estudada em laboratório e em condições de campo, visando avaliar um possível efeito estressor dos derivados vegetais sobre o cupim. Testes de compatibilidade in vitro indicaram resposta dependente da concentração do Nimkol, para a inibição do crescimento de colônias, da produção e da germinação de conídios dos entomopatógenos. A associação de extratos vegetais e entomopatógenos, em geral, diminuiu a atividade fungitóxica sobre H. tenuis. Entretanto, observou-se benefícios da associação de Nimkol (0,2 a 0,4% i.a.) com os isolados de B. bassiana e M. anisopliae quando houve um mínimo de 24 horas de diferença entre a aplicação do produto vegetal e dos fungos no alimento dos cupins. Os resultados de campo para Nimkol, M. asinopliae e a associação de ambos, mostraram tendências semelhantes às observadas para esses tratamentos em laboratório, embora com menores níveis de mortalidade. / The objective of this research was to evaluate the effect of by-products of the Meliaceae Azadirachta indica (neem), Melia azedarach and Trichilia pallida, and of the entomopathogenic fungi Metarhizium anisopliae (isolate 1037) and Beauveria bassiana (isolate 634) in the control of the subterranean termite Heterotermes tenuis. The vegetal by-products were evaluated in relation to the toxicity and repellency for the termite. The commercial formula Nimkol, obtained from neem leaves, was toxic for the termite at concentrations of 0.3% a.i. and higher. There was no detected transmission of the Nimkol from individuals fed with the product to other individuals. The rest of the vegetal extracts were not significantly toxic for H. tenuis. None of the tested products was repellent to the extent of preventing the gallery construction in agar plates or causing them to be closed. In selection experiments on preference, H. tenuis showed repellency towards the aqueous extract of neem leaves (5%). The rest of the by-products tested, including Nimkol-L, were not repellent for the termite. The effect of the vegetal by-products and the entomopathogens, independently and associated, was studied under field and laboratory conditions, looking for a possible stressing effect of the vegetal by-products on the termite. In vitro compatibility tests showed a response of the Nimkol-L, dependent on the concentration, on the growth of colonies, production and germination of conidia of the entomopathogens. The association of entomopathogens and vegetal extracts decreased in general the toxic activity of the fungi over H. tenuis. With a minimal difference of 24h between the application of the vegetal product and the fungi in the food of the termites, a beneficial association was detected between Nimkol-L and isolates of B. bassiana and M. anisopliae. The field results for Nimkol-L, M anisopliae and the combination of the two, showed a trend similar to the one observed in the laboratory results, but with lower mortality levels.

Thesis (MTech (Horticulture))--Cape Peninsula University of Technology, 2018. / Global demand for environmentally-friendly grapevine cultivation and pest control has necessitated an improved understanding of the relationship between soil properties and beneficial naturally occurring antagonists like entomopathogenic fungi (EPF). This group of fungi presents a viable alternative for the control of destructive pests such as the grapevine mealybug. Sixty-six soil samples were collected from 22 vineyards in the Western Cape, South Africa. The association between soil nutrient status and EPF prevalence was then examined. Fungi were isolated with methods of insect baiting and selective media. Fungal strains were identified and characterized using light microscopy and DNA analysis (ITS and BTub). In addition, fungal isolates were tested against a key grapevine pest, Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) using an immersion bioassay at a concentration of 1 x 108 conidia ml-1. Twenty-three fungal strains were isolated and correspondence analysis (CA) of data indicated a positive association between fungal occurrence and moderate to high levels of soil-based macronutrients. Binomial logistic regression analysis revealed that soil N, K, Ca, Mg and S concentrations and C/N ratio were correlated with at least one EPF species. This study showed that some soil nutrient properties correspond to greater occurrence of EPF in grapevine soils. Strains of Beauvaria bassiana (Hypocreales: Clavicipitaceae) caused the highest mortalities (82% to 87%). In chapter three, I examined the effect of B. bassiana inoculation of grape plants on the infestation level of P. ficus, and the growth and volatile constituents of potted grape plants. The grapevines were inoculated with 1 x 108 conidia ml-1 of B. bassiana by drenching before experimentally infesting them with thirty P. ficus adult females. At four weeks post treatments, the fungus was re-isolated from leaves of 50% of the fungus exposed plants. No significant difference (P > 0.05) was observed in all the plant growth parameters measured in the fungus treated and control plants. Plant tissue analysis revealed markedly higher contents of Ca and Mg in leaf tissue of plants exposed to the B. bassiana relative to the control. GC-MS analyses showed that a significantly (X2=5.1; P<0.02) higher number of known anti-insect volatile compounds (9) including napthtalene were present among fungus treated plants compared to the control plants (5). However, B. bassiana did not have any significant effect on total polyphenol, alkaloid and flavonoids. Overall, treatment with fungus did not offer any protection against infestation of P. ficus. In conclusion, this is the first study to report on the isolation of indigenous entomopathogenic fungal (EPF) strains within vineyards of the Western Cape. The study revealed that inoculating grapevine plants during cultivation had a net positive effect on the production of volatile compounds in grapevines. These findings shed light on the mechanisms involved in endophytic fungus-plant-insect interactions. This study contributes valuable information to future development of ecological approaches involving EPF for insect control in vineyards and in general, agricultural settings.

Made available in DSpace on 2017-07-10T17:37:10Z (GMT). No. of bitstreams: 1 Everton Ricardi Lozano da Silva.pdf: 1285547 bytes, checksum: 985ef107fc92d11abb8864885e596668 (MD5) Previous issue date: 2006-03-17 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The objective of this research was evaluation "in vitro" the effect of fungicides and herbicides on Bacillus thuringiensis var. kurstaki (Btk). The five fungicides: epoxiconazole, fentin hydroxide, azoxystrobin, tebuconazole and carbendazim, and five herbicides: sulfentrazone, lactofen, imazethapyr, fenoxaprop and metribuzin were tested at two concentrations, on cells and spores, in nutrient agar (NA) - solid medium (SM), and in nutrient broth (NB) - liquid medium (LM), in addition to their metabolization (MT) by Btk. For cells, the phytosanitary products in NA were tested incorporated into the medium and on filter paper disks impregnated with the products. The evaluation consisted, respectively, of the number of CFU/mL and of the formation of an inhibition halo. For cells, the Bt+products+NB mixture was incubated in Erlenmeyer flasks (30± 2°C, 150rpm, 24h)an then the NB was diluted and inoculated onto NA; the number of CFU/mL was evaluated. The MT and spore tests were performed in liquid medium (water for MT and NB for spores). The effects of the products varied with the category (fungicide and herbicide), their concentrations, technique used, contact time, and developmental stage of the pathogen. The LM test is the most indicated to evaluate the effect of the products on cells. It was observed that most fungicides were compatible with cells, and only epoxiconazole and azoxystrobin were metabolized. The herbicides, independents of the concentrations, were incompatible with cells, and only sulfentrazone and imazethapyr were metabolized. Fentin hydroxide was the only product incompatible with spore / Objetivou-se comparar técnicas para a avaliação in vitro do efeito de fungicidas e herbicidas sobre Bacillus thuringiensis var. kurstaki (Btk). Foram testados cinco fungicidas: epoxiconazole, fentin hidróxido, azoxystrobin, tebuconazole e carbendazin, e cinco herbicidas: sulfentrazone, lactofen, imazethapyr, fenoxaprop e metribuzin, em duas concentrações, sobre células e esporos, em ágar nutriente (AN) - sólido (MS) e caldo nutriente (CN) - líquido (ML), além da metabolização (MT) destes por Btk. Para células, em AN os fungicidas foram testados incorporados ao meio e em discos de papel-filtro impregnados nos produtos. Avaliou-se, respectivamente, as UFC/mL e a formação de halo de inibição. Para células, a mistura Bt+produtos+CN foi incubada em erlemmeyers (30± 2°C, 150rpm, 24h), e em seguida, o meio CN foi diluído e inoculado em AN, avaliando-se o número de UFC/mL. Os testes de MT e esporos foram realizados em líquido (água para MT e CN para esporos). O efeito dos produtos variou conforme a categoria destes (fungicida e herbicida), concentrações, técnica, tempo de contato e fase/estágio do desenvolvimento do patógeno. O teste ML é o mais indicado para a avaliação do efeito em células, sendo observado que a maioria dos fungicidas mostrou-se compatível às mesmas, e apenas epoxiconazole e azoxystrobin foram metabolizados. Os herbicidas, independentes das concentrações, foram incompatíveis com as células, e somente sulfentrazone e imazethapyr foram metabolizados. Fentin hidróxido foi o único produto incompatível com esporos