Thèses sur le sujet « Insects production »

Thesis (Ph. D.)--University of Missouri-Columbia, 2007.
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 February 25, 2008) Vita. Includes bibliographical references.

The species-specific calling songs of male crickets are used by females for species recognition and mate choice. Heritabilities of variation of morphological structures involved in song production, components of the calling song, and body size were estimated for G.firmus. All morphological structures were shown to possess significant additive genetic variation (h$ sp2 sb{ rm S+D} > 0.42)$. One of the five song components examined, pulse rate, was shown to have a significant heritability (h$ sp2 sb{ rm S+D}$ = 0.35). Due to the low correlation between body size and song components, it is unlikely that female G.firmus could use the calling song to assess male body size or wing morph (micropterous or macropterous).

Title from PDF of title page (University of Missouri--Columbia, viewed on Feb 24, 2010). The entire thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file; a non-technical public abstract appears in the public.pdf file. Thesis advisor: Dr. Johannes Schul. Vita. Includes bibliographical references.

In New Zealand Nothofagus forests Ultracoelostoma spp. scale insects produce abundant honeydew which is an important food for native birds, invertebrates, sooty mould, and invasive wasps. Previous models have underestimated honeydew production, potentially because they do not allow for the flow rate of honeydew to vary between insects based on characteristics such as insect size. This research focused on honeydew production rates at the level of the individual insect, how insect characteristics influence production, and whether the strongest predictor of production, ambient air temperature, acts directly on insects or indirectly via effects on trees. Finding out how temperature acts to increase honeydew production will better reveal the physiological processes involved. The study site was Mt. Richardson, Canterbury. In the first part of this study, during March-April 2012, daily mean ambient temperature (range 7.2 – 15.1 °C) had a positive relationship with honeydew production. Insect size positively influenced production at high temperatures, with the largest insects producing 0.296 µg insect⁻¹ h⁻¹ and the smallest insects 0.115 µg insect⁻¹ h⁻¹ at the highest temperature, 15.1 °C. In the second part of this study, during October 2012-January 2013, I manipulated temperature on areas of tree trunk using reflective or clear plastic covers, creating a mean temperature difference of 1.1 °C. However, the effects of tree and insect temperature could not be separated as there was no relationship between either manipulated or ambient temperature and honeydew production. These results show that honeydew production is influenced by individual insect characteristics. This will be important for future models of production. The results also show contradictory effects of temperature on honeydew production, perhaps because of interactions with other unknown factors, which bears further investigation.

Two rates of the insect active ingredient zetacypermethrin (MustangMax) were evaluated for control of summer insect pests on bermudagrass, with application made about one month prior to harvest. Crop was 22 inches tall when treatments were applied and had a dense stand, which also intercepted much of the treatment. Little difference existed between the two rates of zetacypermethrin in this study. Treatments reduced planthopper numbers by slightly over 50% for the first 9 days after application. Reduction of bermudagrass mirid populations was 45% at two days post treatment, but numbers of this insect were numerically higher in MustangMax treated plants than in untreated plots at subsequent sample dates. Treatments also resulted in significantly lower numbers of damsel bugs and minute pirate bugs at two days post treatment, while only reducing grass thrips numbers by about 20% through 13 days after application. Bark lice were more prevalent in the treated plots, thought due to a reduction of predatory beneficial insects. More effectiveness from this chemistry would be expected earlier in the growing season when plants are shorter, therefore allowing greater coverage and contact with insects as this chemistry is not systemic.

The efficacy and field performance of new insecticides for control of insects on vegetables and melons under desert growing conditions has been investigated in small plot trials for the past several years at the Yuma Agricultural Center. Our objective has been to determine how new chemistries will fit into the growers management programs in Arizona. Thus, our research programs have been focused on studies to determine how to integrate these new chemicals into our local management programs in the most cost/effective way possible. This document was created to provide you with an overview of new insecticide chemistries being developed by the Agrichemical Industry for use in vegetables. The first part of this report concisely describes the new types of chemistries being developed The tabular information presented is a summary of the efficacy and activity of the new compounds based on research we have conducted at the Yuma Agricultural Center.

Lectins have long been established as potential agents for use in control of insect pest species. In this study the use of garlic bulb lectins against phloem-feeding hemipteran insect pests is investigated. There is no Bacillus thuringiensis (Et) toxin available which is effective against hemipteran insects. Plant lectins have been shown to be toxic to some insects, including aphids, in particular lectins from garlic. Subsequently heterodimeric (ASAI) and homodimeric (ASAlI) garlic bulb lectins were cloned and e!Cpressed in 31 recombinant yeast system, Pichia pastoris. Recombinant lectins were successfully purified and demonstrated to be functionally active in vitro by haemagglutination assays and toxic to a range of hemipteran insects: pea aphid (Acyrthosiphon pisum), peachpotato aphid (Myzus persicae) and rice brown planthopper (N. lugens) in ~ificial diet bioassays. To analyse the interactions between recombinant ASAII and the gut of A. pisum a pulldown assay was developed to identify specific interactions between ASAII and solubilized A. pisum gut proteins. Proteins which interacted with ASAll were subjected to matrix-assisted laser desoprtion ionization time of flight (MALDI-TOF) mass spectrometric analysis, this revealed that ASAll bound to alanyl aminopeptidase N (alanyl APN), a major constituent glycoprotein of the aphid gut which is rich in mannose oligosaccharides, and membrane-bound sucrase, an enzyme important in the maintenance of osmotic balance in the aphid gut. It was not possible to establish whether transport of ASAll into the haemolymph of A. pisum occurred using a Western blotting approach due to the lack of immunoreactivity of anti-ASA antibodies to the low levels of ASAII expected to be transported. A fusion protein of ASAll-Avidin was created to enable the conjugation of biotinylated peptides of potential insecticidal interest to lectin carrier proteins. One such peptide, leucomyosuppressin (LMS) from cockroach Diploptera punctata, was conjugated to ASAII-Avidin and toxicity to A. pisum was demonstrated in artificial diet bioassays. It was confirmed that avidin is transported into the A. pisum haemolymph, suggesting that if ASAII was not responsible for transport of LMS in to the haemolymph then it may be mediated by avidin. Using a bioinformatic approach a putative A. pisum LMS gene was assembled.

O objetivo do trabalho foi estabelecer as condições ótimas para criação massal de Anagasta kuehniella (Zeller, 1879), definindo a densidade larval por recipiente de criação, associando-a com temperatura e concentração de CO2, produzidos pelo metabolismo larval. Para que este objetivo fosse atingido, foram avaliados: 1) o efeito da temperatura no ciclo evolutivo (ovo-adulto) de A. kuehniella para determinação das suas exigências térmicas; 2) efeito da temperatura nas fases imaturas e nos adultos de A. kuehniella sobre a reprodução, estudando-se o efeito de diferentes temperaturas (na faixa de 18 a 32°C) desde a fase de ovo até a morte dos insetos; efeito de diferentes temperaturas (18 a 32ºC) durante as fases larval e pupal em adultos transferidos para 25°C e o efeito de diferentes temperaturas (18 a 32°C) nos adultos provenientes de imaturos mantidos a 25°C; 3) otimização do número de insetos por recipiente, para criações massais de A. kuehniella, avaliando-se o efeito da densidade de ovos no incremento térmico dos recipientes e na postura, bem como a relação entre incremento térmico x densidade populacional x temperatura de criação na postura de A. kuehniella; 4) produção de dióxido de carbono (CO2) por bandeja de criação de A. kuehniella e o efeito do CO2, na viabilidade do período ovo-adulto e na postura de A. kuehniella. A condição ótima de criação de A. kuehniella foi obtida quando as lagartas no interior das bandejas foram mantidas na temperatura de 25° C. Assim, considerandose o incremento térmico entre 7 e 9°C, gerado pelo metabolismo larval, a temperatura das salas de criação deve ser mantida com temperaturas mais baixas durante o 4° e 5° ínstares do desenvolvimento larval de A. kuehniella. A concentração máxima de CO2, no interior da sala de criação, deve ser inferior a 1.200 ppm, sala esta contendo bandejas com 1 Kg de dieta (97% de farinha de trigo integral e 3% de levedura) inoculadas com 7.200 ovos, pois esta densidade proporcionou uma viabilidade satisfatória do período ovo-adulto (78%), produzindo ainda uma maior quantidade de ovos por bandeja e ovos de A. kuehniella mais pesados.
The objective of this research was to establish the optimum conditions for mass rearing of Anagasta kuehniella (Zeller, 1879), defining the larval density for rearing container, associating it with temperature and concentration of the CO2 produced by the larval metabolism. For this purpose, it was evaluated: 1) the effect of temperature on the life cycle (egg to adult) of A. kuehniella to determine the thermal requirements; 2) effect of temperature in the immature and adults stages of A. kuehniella on reproduction, studying the effect of different temperatures (from 18 to 32°C) from the egg stage until the death of the insects and effect of different temperatures (18 to 32ºC) during the larval and pupal in adults transferred to 25°C and the effect of different temperatures (18 to 32°C) in adults emerging from immature kept at 25°C; 3) optimizing the number of insects per container, for mass rearing of A. kuehniella, evaluating the effect of egg density on temperature increment of containers and oviposition, as well as the relationship between increased temperature X population density X rearing temperature in oviposition of A. kuehniella, 4) carbon dioxide production (CO2) per tray of rearing of A. kuehniella and the effect of CO2 on the viability of the egg-adult and oviposition of A. kuehniella. The optimum condition of rearing A. kuehniella was obtained when the larvae inside the trays were kept at 25°C. Thus, considering the temperature increment between 7 and 9°C, generated by larval metabolism, the temperature in rearing rooms should be kept at lower temperatures during the 4th and 5th instars of the larval development of A. kuehniella. The maximum CO2concentration inside the rearing room should be lower than 1,200 ppm, this room containing trays with 1 kg of diet (97% of whole wheat flour and 3% of yeast) inoculated with 7,200 eggs, as this density provided satisfactory feasibility in the egg-adult period (78%), producing an even greater number of eggs per tray and heavier eggs of A. kuehniella.

Anew chemistry (Spinosad 4SC) was tested for control of beet armyworm and other summers insects in alfalfa. This product did not control beet armyworms as well as the top of label rate of Lorsban 4E, and at one day post treatment had more beet armyworms than the lowest rate of Lorsban tested At three days post treatment both rates of Spinosad 4SC had fewer beet armyworms than the lowest rate of Lorsban tested. Few differences were noted between Spinosad 4SC for beet armyworm control, although fewer alfalfa caterpillars were noted with usage of the higher Spinosad rate, although significantly more beneficial insects were noted at three days post treatment with the lower rate of Spinosad. Lorsban chemistries significantly lowered yellows and hopperburn damage ratings compared with other treatments and a difference was noted between the two Spinosad rates although Empoasca sp. leafhopper numbers were similar.

This thesis looks at the role of wild pollinators in providing services to crops. Two data chapters (2 and 3) are accompanied by a modelling chapter (4) which build on the findings of the field studies. The thesis ends with an overview of the trends in pollinator populations and how these relate to the needs of farmers in the UK (chap-ter 5). It is often assumed that commercial pollinators are appropriate substitutes of wild pollinators on farms; however this view neglects the differing roles that particular pollinator taxa might play in providing pollination services. For example, crops with a long growing system may require multiple pollinators to ensure pollination throughout the season. Strawberries in Scotland have an extremely long growing season, flowering from April to August. Chapter 2 presents a study showing season-al complementarity between different pollinating taxa across strawberry farms in Scotland. Pollinators of strawberries also differed in their responses to weather pa-rameters indicating that preserving multiple pollinator taxa could ensure yields un-der different weather scenarios. The requirements of a long-growing season and ad-verse weather may be specific to strawberry production in Scotland, but the valua-tion of multiple taxa can be generalised to systems with differing needs, and also to different ecosystem services. Wild bees are not only valuable for providing complementary services to commercial pollinators, but are also valuable in the longer term as it is unknown whether com-mercial pollinators will be available in the future. There are threats to the supply of honeybees which have already triggered price rises; such supply shocks could force farmers to leave production or to seek other ways of providing pollination, including supporting wild pollinators. However farm management pressures, in particular pes-ticide use, could threaten the ability of wild pollinators to continue to support crop production. The interplay of pesticides and pollination is discussed in chapter 3 and 4. Chapter 3 presents an experiment undertaken on soft-fruit farms which had and had not used the neonicotinoid, thiacloprid, and shows that nests exposed to thia-cloprid had higher worker mortality, and lower male production than those at con-trol farms. This has implications both for pollination services now, as worker mor-tality will reduce the number of bees visiting farms, and also for the maintenance of future pollination services through decreased reproductive capacity of exposed nests. Chapter 4 uses a theoretical model to link pesticide use and habitat use to pollina-tion services, and shows that the use of commercial pollinators could mask the de-cline in wild populations, making local extinctions more likely. Chapter 5 sets out the status and extent of pollinators in the UK, along with popu-lation trends, trends in habitat and trends in pesticide use to provide an overview of how well pollination services are likely to meet the ongoing needs of crop farmers.

Food habits, net-spinning activity, energetics, and mercury accumulation in Hydrospsyche morosa were examined over a one year period on the South River in central Virginia. Feeding nets were observed as early as April and were widespread by May. Nets were virtually absent from late November through March. Gut content analysis revealed seasonal patterns in the consumption of various food items. From April through October, when feeding nets were widespread, detritus formed the bulk of the diet in terms of both numbers of particles and volume occupied. From November through March however, the algal component dominated in terms of numbers of particles although the detritus component still occupied a greater volume. Ivlev's preference index was employed and indicated that the seasonal differences in the relative amount of the three food types were not simply a matter of changing seston concentrations, but rather suggested a shift from a filter-feeding mode of feeding in the summer months to grazing on diatoms in the winter. H. morosa was bivoltine on the South River. The estimate of secondary production for the summer cohort was 3,246 mg AFDW/m²/yr, while the estimate for the winter cohort was 2,145 mg AFDW/m²/yr. The secondary production also was estimated for each season based on food habits to determine the impact of the observed seasonal switch in feeding habits on production and egestion rates. During the summer, the detritus component contributed most to production averaging about 50 percent. Animal and algal material contributed 30 and 20 percent, respectively. During the winter, algal material contributed most to the production, averaging just over 62 percent. Detritus also contributed during the winter averaging over 30 percent. Monthly rates of production and egestion were between 3 and 3.5 times faster during the summer. The concentrations of total mercury in seston, periphyton, and in the body tissue of H. morosa were analyzed each month. Mercury concentrations were between four and six times higher in the seston than in the periphyton. The concentration of mercury in the body tissue of H. morasa ranged from 0.14 ppm in March to over 1.20 ppm in July. Differences in Mercury concentration in the insects between seasons were significant. Regression analysis revealed a significant relationship between Hg concentration in the insects and the relative amount of detritus found in the guts.
Master of Science

The capsid protein of Helicoverpa armigera stunt virus (HaSV) a T=4 insect virus was expressed in Spodoptera frugiperda 9 cells using a baculovirus vector. When the insect cells were infected at a high MOl the expressed coat protein assembled into virus-like particles (VLPs) that spontaneously underwent maturation and were morphologically indistinguishable from wild-type HaSV. The VLPs were electron dense when viewed under EM and encapsidated their coat protein mRNA. When Sf9 cells were infected at a low multiplicity of infection (MOl) the expressed capsid protein assembled into procapsids that did not spontaneously undergo maturation. These procapsids underwent autoproteolytic maturation cleavage when they were treated with an acidic buffer. The procapsids were used in the encapsidation of a FITC labelled peptide. The peptide encapsidating VLPs showed an increase in their buoyant density that was not collaborated by an increase in the concentration of the FITC labelled peptide detected when these samples were compared to control samples with similar buoyant densities.

A series of experiments were conducted to determine the potential of a new method of steam treatment for the control of soil-borne diseases, insects and weeds in sustainable crop production.  The new method involved rapidly heating a bed of prepared soil on a porous base by passing an upward flow of steam through it.  The aim was to determine whether the new method would be suitable for use in a field machine, making large-scale field steaming practically and economically viable. In the first experiments, the feasibility of the new method was tested.  It was shown that it was an effective and rapid way in which a soil bed could be steamed.  However, for some soils at, or near, permanent wilting point, the entrainment of aggregates in the steam flow was shown to be a problem.  The second series of experiments showed that the key factor determining the flow rate at which aggregate entrainment occurred was the mean aggregate diameter of the soil being treated.  The third series of experiments examined the rate at which heated soil would cool when placed in the field.  It was shown that where there was contact with unheated field soil, cooling was very rapid.  The final series of experiments investigated the effects of the new steaming method on the soil.  A three minute steaming time was used to account for the short time it had been shown some of the heated soil would remain at steam temperature when replaced in the field.  The effects of the new method, including the effectiveness of disinfection, were shown to be similar to those of a conventional steam treatment. It was concluded that the new steaming method was an effective way to steam treat soil and should be suitable for use in a field machine.

Thesis (Ph. D.)--University of Missouri-Columbia, 2007.
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 January 31, 2008) Vita. Includes bibliographical references.

Thesis (Ph. D.)--University of Missouri-Columbia, 2007.
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 October 18, 2007) Vita. Includes bibliographical references.

Com o objetivo de incentivar novos pesquisadores e empreendedores da área de controle biológico de pragas, foi realizada uma série de experimentos voltados à qualidade da produção massal do agente biológico Chrysoperla externa (NEUROPTERA; CHRYSOPIDAE). Estudaram-se aspectos biológicos desta espécie, com o interesse de se delinear um sistema de produção eficiente, baseado em 7 módulos de gaiolas de adultos, em que se coletam ovos por 49 dias. Para se manter uma produtividade constante, esses módulos são montados com uma diferença de tempo de 7 dias entre eles. Outras técnicas e análises de custos de produção também estão descritas. Realizou-se ainda um estudo do efeito de diferentes doses do anticontaminante nipagin, sobre a produtividade dos adultos, evidenciou que sua utilização em biofábricas de C. externa não traz benefícios significativos. Com esse trabalho é possível produzir ovos de C. externa com maior eficiência.
This research deals with biological aspects and their influence on the quality of Chrysoperla externa produced in laboratory. The system of production was based on seven modules of adult cages from which one collected eggs during 49 days. Such modules must be prepared at each seven days to provide a constant flowing production. Other techniques and analysis of costs of production are described as well. A study of different concentrations (dosis) of the anticontaminant nipagin on C. externa adults indicated that its use showed no significant benefits for the production of this predator.

To refine our knowledge and to adequately test hypotheses concerning theoretical and applied aspects of invasion biology, successful and unsuccessful invaders should be compared. This study investigated insect establishment patterns by comparing the climatic preferences and biological attributes of two groups of polyphagous insect species that are constantly intercepted at New Zealand's border. One group of species is established in New Zealand (n = 15), the other group comprised species that are not established (n = 21). In the present study the two groups were considered to represent successful and unsuccessful invaders. To provide background for interpretation of results of the comparative analysis, global areas that are climatically analogous to sites in New Zealand were identified by an eco-climatic assessment model, CLIMEX, to determine possible sources of insect pest invasion. It was found that south east Australia is one of the regions that are climatically very similar to New Zealand. Furthermore, New Zealand shares 90% of its insect pest species with that region. South east Australia has close trade and tourism links with New Zealand and because of its proximity a new incursion in that analogous climate should alert biosecurity authorities in New Zealand. Other regions in western Europe and the east coast of the United States are also climatically similar and share a high proportion of pest species with New Zealand. Principal component analysis was used to investigate patterns in insect global distributions of the two groups of species in relation to climate. Climate variables were reduced to temperature and moisture based principal components defining four climate regions, that were identified in the present study as, warm/dry, warm/wet, cool/dry and cool/moist. Most of the insect species established in New Zealand had a wide distribution in all four climate regions defined by the principal components and their global distributions overlapped into the cool/moist, temperate climate where all the New Zealand sites belong. The insect species that have not established in New Zealand had narrow distributions within the warm/wet, tropical climates. Discriminant analysis was then used to identify which climate variables best discriminate between species presence/absence at a site in relation to climate. The discriminant analysis classified the presence and absence of most insect species significantly better than chance. Late spring and early summer temperatures correctly classified a high proportion of sites where many insect species were present. Soil moisture and winter rainfall were less effective discriminating the presence of the insect species studied here. Biological attributes were compared between the two groups of species. It was found that the species established in New Zealand had a significantly wider host plant range than species that have not established. The lower developmental threshold temperature was on average, 4°C lower for established species compared with non-established species. These data suggest that species that establish well in New Zealand have a wide host range and can tolerate lower temperatures compared with those that have not established. No firm conclusions could be drawn about the importance of propagule pressure, body size, fecundity or phylogeny for successful establishment because data availability constrained sample sizes and the data were highly variable. The predictive capacity of a new tool that has potential for eco-climatic assessment, the artificial neural network (ANN), was compared with other well used models. Using climate variables as predictors, artificial neural network predictions were compared with binary logistic regression and CLIMEX. Using bootstrapping, artificial neural networks predicted insect presence and absence significantly better than the binary logistic regression model. When model prediction success was assessed by the kappa statistic there were also significant differences in prediction performance between the two groups of study insects. For established species, the models were able to provide predictions that were in moderate agreement with the observed data. For non-established species, model predictions were on average only slightly better than chance. The predictions of CLIMEX and artificial neural networks when given novel data, were difficult to compare because both models have different theoretical bases and different climate databases. However, it is clear that both models have potential to give insights into invasive species distributions. Finally the results of the studies in this thesis were drawn together to provide a framework for a prototype pest risk assessment decision support system. Future research is needed to refine the analyses and models that are the components of this system.

The overall objective of this study was to test the hypothesis that insects can vector F. tumidum conidia to infect gorse plants with the aim of developing an alternative approach to mycoherbicide delivery to control weeds. Four potential insect species (Apion ulicis, Cydia ulicetana, Epiphyas postvittana and Sericothrips staphylinus) were assessed for their ability to vector F. tumidum conidia. To achieve this, the external microflora (bacteria and fungi) and the size and location of fungal spores on the cuticle of these insect species were determined. In addition, the ability of the insects to pick up and deposit F. tumidum conidia on agar was studied. Based on the results from these experiments, E. postvittana was selected for more detailed experiments to determine transmission of F. tumidum to infect potted gorse plants. The factors promoting pathogenicity of F. tumidum against gorse and the pathogen loading required to infect and kill the weed were also determined. The external microflora of the four insect species were recovered by washing and plating techniques and identified by morphology and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and sequencing of internally transcribed spacer (ITS) and 16S rDNA. A culture-independent technique (direct PCR) was also used to assess fungal diversity by direct amplification of ITS sequences from the washings of the insects. All insect species carried Alternaria, Cladosporium, Nectria, Penicillium, Phoma, Pseudozyma spp. and entomopathogens. Ninety four per cent of the 178 cloned amplicons had ITS sequences similarity to Nectria mauritiicola. E. postvittana carried the largest fungal spores (mean surface area of 125.9 ìm2) and the most fungal CFU/insect. About 70% of the fungi isolated from the insects were also present on the host plant (gorse) and the understorey grass. The mean size of fungal spores recovered from the insect species correlated strongly with their body length (R² = 85%). Methylobacterium aquaticum and Pseudomonas lutea were common on all four insect species. Pseudomonas fluorescens was the most abundant bacterial species. In the pathogenicity trials, the effectiveness of F. tumidum in reducing root and shoot biomass of 16 and 8 wk old gorse plants was significantly increased with wounding of the plants. Older plants (32 wk old) which were wounded and inoculated were significantly shorter, more infected and developed more tip dieback (80%) than plants which were not wounded (32%). This indicates that damage caused by phytophagous insect species present on gorse through feeding and oviposition may enhance infection by F. tumidum. Wounding may release nutrients (e.g. Mg and Zn) essential for conidia germination and germ tube elongation and also provide easier access for germ tube penetration. Conidial germination and germ tube length were increased by 50 and 877%, respectively when incubated in 0.2% of gorse extract solution for 24 h compared with incubation in water. Inoculum suspensions amended with 0.2% of gorse extract caused more infection and significantly reduced biomass production of 24 wk old gorse plants than suspensions without gorse extract. A minimum number of about 900 viable conidia/infection site of F. tumidum were required to infect gorse leaves. However, incorporation of amendments (which can injure the leaf cuticle) or provision of nutrients (i.e. gorse extract or glucose) in the formulation might decrease the number of conidia required for lesion formation. Scanning electron micrographs showed that germ tube penetration of gorse tissue was limited to open stomata which partly explain the large number of conidia required for infection. The flowers and leaves were more susceptible to F. tumidum infection than the spines, stems and pods. An experiment to determine the number of infection sites required to cause plant mortality showed that the entire plant needs to be inoculated in order for the pathogen to kill 10 wk old plants as F. tumidum is a non systemic pathogen. The number of infection sites correlated strongly with disease severity (R² = 99.3%). At least 50% of the plant was required to be inoculated to cause a significant reduction in shoot dry weight. F. tumidum, applied as soil inoculant using inoculated wheat grains in three separate experiments, significantly suppressed gorse seedling emergence and biomass production. In experiments to determine the loading capacity of the insect species, E. postvittana, the largest insect species studied, carried significantly more (68) and deposited significantly more (29) F. tumidum conidia than the other species. Each E. postvittana, loaded with 5,000 conidia of F. tumidum, transmitted approximately 310 conidia onto gorse plants but this did not cause any infection or affect plant growth as determined by shoot fresh weight and shoot height. E. postvittana on its own did not cause any significant damage to gorse and did not enhance F. tumidum infection. It also failed to spread the pathogen from infected plants to the healthy ones. There was no evidence of synergism between the two agents and damage caused by the combination of both E. postvittana and F. tumidum was equivalent to that caused by F. tumidum alone. This study has shown that E. postvittana has the greatest capacity to vector F. tumidum since it naturally carried the largest and the most fungal spores (429 CFU/insect). Moreover, it naturally carried Fusarium spp. such as F. lateritium, F. tricinctum and Gibberella pulicaris (anamorph Fusarium sambucinum) and was capable of carrying and depositing most F. tumidum conidia on agar. Coupled with the availability of pheromone for attracting the male insects, E. postvittana may be a suitable insect vector for delivering F. tumidum conidia on gorse using this novel biocontrol strategy. Although it is a polyphagous insect, and may visit non-target plants, F. tumidum is a very specific pathogen of gorse, broom and a few closely related plant species. Hence, using this insect species to vector F. tumidum in a biological control programme, should not pose a significant threat to plants of economic importance. However, successful control of gorse using this "lure-load-infect" concept would depend, to a large extent on the virulence of the pathogen as insects, due to the large size of F. tumidum macroconidia, can carry only a small number of it.

Questa tesi è frutto della collaborazione tra l'Università e Lofarma S.p.A., un’azienda farmaceutica italiana leader nel settore che produce preparati per pazienti allergici come kit diagnostici e immunoterapie. A tale scopo nel reparto di Acarologia ogni anno vengono allevati e raccolti decine di chilogrammi di acari adulti e, dopo alcune manipolazioni, utilizzati nel reparto produttivo come materia prima per la maggior parte delle formulazioni. Lo scopo di questo progetto è analizzare l'attuale metodo di produzione e studiare se alcuni passaggi potrebbero essere ottimizzati per migliorare la resa, il tasso di produzione e la qualità della materia prima, cercando nel contempo di ridurre costi e tempi di lavorazione. La ricerca è stata suddivisa in 2 aree principali: 1) Procedure di Allevamento (valutazione della qualità della dieta e del ceppo allevato) e 2) Manipolazioni della Materia Prima (ottimizzazione del processo di pulizia e valorizzazione della materia prima). Tra questi, i risultati più significativi sono stati raggiunti nella sezione Manipolazione delle Materie Prime, dove viene descritto un nuovo processo di rifinitura in grado di ottenere rese finali più elevate in tempistiche più brevi. Dopo aver analizzato l'intero ciclo produttivo, è possibile concludere che, nel contesto del Reparto di Acarologia, è più conveniente procedere con una migliore manipolazione della materia prima piuttosto che modificare le attuali metodiche di allevamento, che sembrano già adatte per le esigenze di Lofarma.
This thesis is based on collaboration between University and Lofarma S.p.A., a leading Italian pharmaceutical company which produce preparation for allergic patients like diagnostic kits and immunotherapies. To this purpose every year dozens of kilograms of adult mites are reared and collected in the Acarology department and, after manipulations, used in the Production Department as raw material for most of the preparation. The aim of this project is to analyze the current production methodology of Acarology department and investigate if some steps could be optimized to improve the yield, the production rate and the quality of the raw material while trying to reduce costs and processing times. The research has been divided in 2 main areas: 1) Rearing Procedures (quality assessment about the diet and the strain enacted) and 2) Raw Material Manipulations (optimization of the refining process and valorization of the Raw Material). Between those, most significant results have been achieved in the Raw Material Manipulation section, where is described a novel refining process capable of obtaining higher final yields in a shorter working time. After analyzing the whole manufacturing cycle, is possible to conclude that, within the context of the Acarology Department, is more convenient to proceed with a better manipulation of the raw material in the refining process rather than modifying the actual rearing procedure, which is already suitable for Lofarma needs.

Na cultura da soja ocorre um grupo de percevejos sugadores de sementes, Euschistus heros (E.), Piezodorus guildinii (West.) e Nezara viridula (L.), que causam diversos distúrbios fisiológicos durante o cultivo, como o atraso da maturidade fisiológica, a retenção foliar, as perdas na produtividade e a redução do potencial fisiológico das sementes. A reação dos genótipos de soja ao estresse por percevejos é uma abordagem importante durante a etapa de melhoramento, produção de sementes e desenvolvimento de novas linhagens de soja resistente. O objetivo deste estudo foi comparar os parâmetros de produção e qualidade das sementes, e a resposta de defesa da planta dos diferentes genótipos de soja em condições de ataque de percevejos. Os ensaios foram conduzidos na Estação Experimental de Anhumas (Departamento da Genética, USP, ESALQ), com as populações de plantas de soja submetidas à condição de infestação natural dos percevejos nos anos agrícolas 2012/13 e 2013/2014, com plantas cultivadas sem o controle químico de percevejos e com o controle, em blocos casualizados com cinco repetições em cada sistema de controle. Análises dos dados foram feitas com ANOVA e MANOVA. No primeiro ano, vinte quatro genótipos de soja foram avaliados para os parâmetros agronômicos e de resistência da planta (período de formação de sementes (PFS), ciclo das plantas (Ciclo), altura da planta (AP), índice de acamamento (IA), valor agronômico (VA), número de vagem (NV), peso de mil sementes (PMS), produtividade (PROD), retenção foliar (RF) e peso de sementes boas (PSB)). No segundo ano com oito genótipos de soja, as plantas foram avaliadas pelos mesmos parâmetros agronômicos e de resistência do primeiro ano, exceto o NV e com o acréscimo da avaliação do índice percentual de danos nas vagens (IPDV) e as sementes quanto a qualidade fisiológica (germinação, envelhecimento acelerado, emergência, condutividade elétrica e tetrazólio). Além disso, as sementes e vagens de dois genótipos LQ1050 e CD215, coletadas entre os estágios R5 e R6, foram avaliadas quanto ao teor de isoflavonas em condições de estresse por percevejo no campo. Com o monitoramento da população de percevejos foi possível verificar um aumento de percevejos no PFS, sendo que a densidade populacional no ano agrícola 2012/13 foi mais alta. Para a seleção efetuada no primeiro ano agrícola 2012/13, considerou-se como principal critério o PSB mínimo e máximo para genótipos resistentes e suscetível ao percevejo para a avaliação da qualidade das sementes no segundo ano de cultivo 2013/14. Genótipos de soja com alta produtividade não garantem que os mesmos tenham resistência ao complexo de percevejos e produzam sementes com alta qualidade fisiológica. Foi verificada, nos oitos genótipos de soja do segundo ano de cultivo, variabilidade genética entre cultivares e linhagens de soja para características de qualidade fisiológica avaliada por meio dos testes de germinação e de vigor. Parâmetros utilizados para avaliação da qualidade fisiológica podem ser correlacionados com a resistência da planta ao complexo de percevejos. Coumestrol e gliocelina, compostos fitoalexinas, foram determinados nas vagens de genótipos contrastantes, mas não nas sementes.
A group of stink bugs composed by Euschistus heros (E.), Piezodorus guildinii (West.), and Nezara viridula (L.) causes several physiological disturbs in soybean during the field production, such as: delayed physiological maturity, leaf retention, yield loss, and decreased seed quality and germination potential. The reaction of soybean genotypes to stink bugs complex is an important approach for the crop breeding, seed production, and development of new resistant lines. This work aimed to compare the production and seed quality parameters, besides the plant defense under conditions of stink bugs attack. Assays were carried out at the Experimental Station of Anhumas (Department of Genetics, USP, ESALQ) with the soybean plants submitted to natural infestation by stink bugs, at 2012/13 and 2013/14 crop seasons, with absence and presence of chemical control of the insects, in a randomized block design with five replicates. The data were submitted to both ANOVA and MANOVA analysis. At the first season, 24 genotypes were evaluated as for the agronomic and plant resistance traits: seed formation period, plant life cycle, plant height, lodging index, agronomic value, number of pods per plant, mass of 1000 seeds, yield, foliar retention, and mass of healthy seeds. At the second season, eight genotypes were evaluated by the same agronomic and plant resistance traits of the first season, except the number of pods per plant. The pods damage index and seed physiological quality (germination, accelerated aging, emergence, electrical conductivity, and tetrazolium test) traits were evaluated only in the second season. The pods and seeds of LQ1050 and CD215 genotypes were collected between the R5 and R6 stages and had the isoflavones levels evaluated in conditions of stress by stink bugs attack. Through the monitoring of stink bugs population, it was possible to notice the increase of the insects in the seed formation period, with higher population density in the 2012/13 season. In the first season, the minimum and maximum masses of healthy seeds were taken as the main criterion to select susceptible and resistant genotypes, respectively, to the stink bugs attack, for the seed quality evaluation in the second season 2013/14. High yield genotypes do not ensure resistance to the stink bugs complex and, therefore, seeds with high physiological quality. We have noticed the genetic variability among the genotypes as for the physiological quality evaluated through the germination and vigor tests, at the second season. Traits used for the physiological quality can be correlated with the plant resistance to the stink bugs complex. Coumestrol and glyceollin, phytoalexin compounds, were identified in pods of the contrasting genotypes, but not in the seeds.

Boston University. University Professors Program Senior theses.
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2031-01-02

Les virus-like particles (VLPs) han sorgit com a una alternativa a les vacunes convencionals basades en virus atenuats o inactivats. La seva capacitat d’autoacoblament en base a l’expressió d’una proteïna matriu i l’absència de material genòmic d’origen víric les fa candidats atractius per a una multitud d’aplicacions. Les VLPs de Gag del virus de la immunodeficiència humana (VIH) són un tipus de VLPs amb envolta que han atret especial interès degut a les seves propietats estructurals, amb aplicacions en teràpia gènica, nanotecnologia i el desenvolupament de vacunes multivalents. Les línies cel·lulars d’insecte són un sistema de referència per a produir aquest tipus de nanopartícules ja que proporcionen les condicions adients per la seva producció i assemblatge. En aquesta tesi s’ha avaluat la producció de VLPs del VIH-1 en les línies d’insecte Sf9 i High Five amb el sistema d’expressió baculovirus i transfecció transitòria. Per tal d’assolir aquest objectiu, s’ha emprat una aproximació basada en l’ús combinat de metodologies de disseny d’experiments i funcions de resposta combinada. Paral·lelament, s’han incorporat una sèrie de tècniques de mesura per tal de monitoritzar i quantificar el procés productiu i per a la caracterització final de les VLPs. En el primer capítol s’analitzen les característiques d’ambdues línies cel·lulars d’insecte com a plataformes per a la producció de VLPs de GageGFP amb el sistema d’expressió baculovirus. En tots dos casos, l’observació de les VLPs per microscòpia electrònica de criogènia permet determinar que tenen una mida similar i també permet detectar la presència d’altres poblacions de nanopartícules. L’anàlisi dels nivells de producció de baculovirus resulta en un increment de 23 vegades de virus infectius en les cèl·lules Sf9 mentre que una proporció més gran de virus d’oclusió s’observa en les cèl·lules High Five. La presència d’aquest últim fenotip de baculovirus evidencia un canvi en la complexitat de la línia cel·lular High Five després de la infecció amb el baculovirus. Finalment, la combinació de les tècniques d’ultracentrifugació i virometria de flux mostra que les VLPs derivades de High Five tenen un major coeficient de sedimentació, la qual cosa indica que aquestes VLPs poden estar associades amb altres elements cel·lulars. Al segon capítol es determinen les condicions òptimes per a la producció de VLPs en les cèl·lules Sf9 i High Five amb el sistema d’expressió baculovirus. En aquest sentit, s’apliquen metodologies de disseny d’experiments i funcions d’optimització amb tècniques de quantificació directa de nanopartícules per tal d’aprofundir en aquests sistemes. Inicialment s’aborden dues situacions objectiu, la primera cercant la maximització de la concentració de VLPs (Quantitat) i la segona buscant un balanç entre producció i percentatge de VLPs amb estructura completa (Qualitat). Los niveles de producción final de VLPs en la condición de calidad son 4.5 veces más elevados para las células Sf9 mientras que en la condición de cantidad se obtienen concentraciones de VLPs similares para ambas líneas. Al tercer capítol de la tesi es desenvolupa una estratègia de producció lliure de baculovirus i basada en la transfecció transitòria de ADN plasmídic amb polietilenimina (PEI). Anàlogament al capítol 2, s’implementa una aproximació sistemàtica de disseny d’experiments i funcions d’optimització. En ambdós casos, el recanvi de medi abans de la transfecció resulta ser beneficiós per tal d’assolir els nivells més alts d’expressió. Les condicions òptimes de concentració de cèl·lules viables, ADN i PEI es determinen en aquest estudi i la formació correcta de les VLPs produïdes es corrobora per microscòpia electrònica de criogènia. En aquest cas, les cèl·lules Sf9 assoleixen un increment de 8.4 vegades en la producció de VLPs respecte a la línia cel·lular High Five. A l’últim capítol de la tesi es desenvolupen conjunts de cèl·lules Sf9 i High Five amb expressió estable i contínua de VLPs al llarg del temps. Aquests conjunts de cèl·lules d’expressió estable es generen a partir de la integració aleatòria d’ADN codificant al genoma de les cèl·lules, i les que són més productives se seleccionen per citometria en base a la seva fluorescència. En relació a la producció de VLPs, s’aconsegueix un increment de 3.7 vegades en les cèl·lules High Five respecte les Sf9. Finalment, l’estabilitat d’aquests grups cel·lulars d’expressió estable es corrobora al llarg d’un mes en cultiu.
Las virus-like particles (VLPs) han surgido como a una alternativa a las vacunes convencionales basadas en virus atenuados o inactivados. Su capacidad de autoensamblaje en base a la expresión de una proteína matriz y la ausencia de material genómico de origen vírico las hace candidatos atractivos para una multitud de aplicaciones. Las VLPs de Gag del virus de la inmunodeficiencia humana (VIH) son un tipo de VLPs con envuelta que ha suscitado especial interés debido a sus propiedades estructurales, con aplicaciones en terapia génica, nanotecnologia y el desarrollo de vacunas multivalentes. Las líneas celulares de insecto son un sistema de referencia para producir este tipo de nanopartículas puesto que proporcionan les condiciones adecuadas para su producción y ensamblaje. En esta tesis se ha evaluado la producción de VLPs del VIH-1 en las líneas de insecto Sf9 y High Five con el sistema de expresión baculovirus y transfección transitoria. Para conseguir este objetivo, se ha utilizado una aproximación basada en el uso combinado de metodologías de diseño de experimentos y funciones de respuesta combinada. Paralelamente, se han incorporado una serie de técnicas de medición para monitorizar y cuantificar el proceso productivo y para la caracterización final de las VLPs. En el primer capítulo se analizan las características de ambas líneas celulares de insecto como plataformas para la producción de VLPs de GageGFP con el sistema de expresión baculovirus. En ambos casos, la observación de las VLPs mediante microscopia electrónica de criogenia permite determinar que tienen un tamaño similar y también permite detectar la presencia de otras poblaciones de nanopartículas. El análisis de los niveles de producción de baculovirus resulta en un incremento de 23 veces de virus infectivos en las células Sf9 mientras que una proporción más gran de virus de oclusión se observa en las células High Five. La presencia de este último fenotipo de baculovirus evidencia un cambio en la complejidad de la línea celular High Five después de la infección con el baculovirus. Finalmente, la combinación de les técnicas de ultracentrifugación y virometría de flujo muestran que las VLPs derivadas de High Five tienen un mayor coeficiente de sedimentación, lo que indica que éstas pueden estar asociadas con otros elementos celulares. En el segundo capítulo se determinan las condiciones óptimas para la producción de VLPs en las células Sf9 y High Five con el sistema de expresión baculovirus. En este sentido, se aplican metodologías de diseño de experimentos y funciones de optimización con técnicas de cuantificación directa de nanopartículas para profundizar en estos sistemas. Inicialmente se consideran dos situaciones objetivo, la primera investiga la maximización de la concentración de VLPs (Cantidad) y la segunda busca un balance entre producción y porcentaje de VLPs ensambladas (Calidad). Los niveles de producción final de VLPs en la condición de calidad son 4.5 veces más elevados para las células Sf9 mientras que en la condición de cantidad se obtienen concentraciones de VLPs similares para ambas líneas. En el tercer capítulo de la tesis se desarrolla una estrategia de producción libre de baculovirus y basada en la transfección transitoria de ADN plasmídico con polietilenimina (PEI). Análogamente al capítulo 2, se implementa una aproximación sistemática de diseño de experimentos y funciones de optimización. En ambos casos, el recambio de medio previo a la transfección resulta ser beneficioso para conseguir los niveles más altos de expresión. Las condiciones óptimas de concentración de células viables, ADN y PEI se determinan en este estudio y la formación correcta de las VLPs producidas se corrobora por microscopia electrónica de criogenia. En este caso, las células Sf9 consiguen un incremento de 8.4 veces en la producción de VLPs respecto a la línea celular High Five. En el último capítulo de la tesis se desarrollan grupos de células Sf9 y High Five con expresión estable y continua de VLPs a lo largo del tiempo. Estos conjuntos celulares de expresión estable se generan a partir de la integración aleatoria de ADN codificante en el genoma de las células, y las que son más productivas se seleccionan por citometría en base a su fluorescencia. En cuanto a la producción de VLPs, se consigue un incremento de 3.7 veces en las células High Five respecto a las Sf9. Finalmente, la estabilidad de estos grupos celulares de expresión estable se corrobora a lo largo de un mes en cultivo.
Virus-like particles (VLPs) have emerged as an interesting alternative to conventional vaccines based on live-attenuated or inactivated viruses. Their capacity for self-assembling upon expression of the core protein and the lack of viral genomic material make them excellent candidates for a variety of purposes. Gag VLPs from the human immunodeficiency virus (HIV) are a type of enveloped VLPs that have drawn special attention due to their structural properties with applications in gene therapy, nanobiotechnology and multivalent vaccine development. Insect cell lines are a reference system to produce these types of nanoparticles since they provide the ideal conditions for their production and assembly. In this work, the production of HIV-1 GageGFP VLPs is assessed in Sf9 and High Five insect cells with the baculovirus expression vector system (BEVS) and transient gene expression (TGE). A rational approach based on the combination of Design of Experiments (DoE) and desirability functions is used to optimize the VLP production conditions. Advanced measurement techniques are implemented to monitor and quantify the production process and for final VLP characterization. In the first chapter, the characteristics of both insect cell lines as platforms for GageGFP VLP production with the BEVS are analyzed. In both cases, similar VLP sizes for both cells are measured by cryogenic electron microscopy (cryo-EM) and other nanoparticle populations are identified. The analysis of baculovirus production levels results in a 23-fold increase of budded virus in Sf9 cells while a larger amount of occlusion-derived virus is detected in High Five cells. The presence of this baculovirus phenotype evidences a shift in the cellular complexity of High Five cells upon baculovirus infection. Finally, the combination of analytical ultracentrifugation with flow virometry reveals a higher sedimentation coefficient for High Five-derived VLPs, indicating their possible association with other cellular compounds. In the second chapter, the optimal conditions for VLP production in Sf9 and High Five cells with the BEVS are determined by means of DoE and desirability functions. Different methodologies based on direct nanoparticle quantification are used to gain insight into these systems. Two objective situations are defined, one targeting the maximization of the VLP titer (Quantity) and the second one aiming to find a balance between production and assembled VLPs (Quality). Final VLP production levels in the quality condition are 4.5-fold higher for Sf9 cells while similar VLP concentrations are found for both insect cells in the quantity condition. In the third chapter of this thesis, a baculovirus-free VLP production strategy is optimized for both insect cells based on plasmid-mediated TGE with polyethylenimine (PEI). As in chapter 2, a systematic approach combining DoE and desirability functions is implemented. In both cases, medium exchange before transfection proves to be beneficial to achieve the highest transgene expression yields. Then, the optimal conditions for viable cell concentration at transfection, DNA and PEI concentrations are determined and the correct formation of the VLPs produced is corroborated using cryo-EM. In this case, Sf9 cells achieve a 8.4-fold increase in VLP production compared to High Five cells. In the last chapter, stable Sf9 and High Five cell pools to produce VLPs are developed by random integration and selection of the high producer cells using fluorescence-activated cell sorting. In terms of VLP production, a 3.7-fold increase in VLP titer is achieved in High Five over Sf9 stable pools. Finally, cell pool stability is successfully corroborated during the course of a month.

Denna rapport är en jämförande studie av utvalda konsolidanter som används till att stabilisera nedbrutet insektsangripet trä. Av de trägnagande skadeinsekter vi har i Sverige är det främst den strimmiga trägnagaren som är intressant i möbelsammanhang. I rapporten beskrivs den strimmiga trägnagaren, en liten skadeinsekt som kan åstadkomma stor skada om den får verka ostörd. Insekten finns utspridd i hela Europa och är ett stort problem då den angriper alla typer av träslag. Nedbrutet insektsangripet trä förlorar sin styrka och kan behöva konsolideras genom att injicera konsolidanter in i träet. Studien är tänkt att jämföra styrkan på de vanligast förkommande och använda konsolidanterna. Urval av dessa har dels gjorts genom att intervjua konservatorer i Sverige, Norge och Danmark, dels utifrån tillgänglig litteratur och forskning. I studien injiceras de utvalda konsolidanterna in i provbitar av artificiellt konstruerat insektsangripet trä som sedan utsätts för hållfasthetsprover. Provbitarna är utformade i kuber av björk, 45x45x45 mm och är borrade med 2 mm stora hål för att efterlikna angripet trä. Två olika typer av hållfasthetsprov har använts för att mäta styrkan hos de olika konsolidanterna, tryckhållfasthet tvärsfiber och skjuvhållfasthet. Resultaten av hållfasthetsproverna visar inga tydliga tecken på att de injicerade konsolidanterna gett någon direkt styrka till provbitarna. I tryckhållfasthets tvärsfiber finns en dock en tendens tillförhöjda värden. För att kunna dra någon slutsats bör ytterligare studier genomföras.

This report is a comparative study of a chosen set of consolidating agents used to stabilize wood suffering from the infestation of wood boring insects. When considering furniture, the most common wood boring insect in Sweden is the Furniture Beetle. This report discusses the Furniture Beetle, a small insect which if left alone, achieves a great amount of damage. This particular beetle can be found in all of Europe and is a sizable problem, since it can attack all species of wood. Infested wood is destabilized by the beetles boring a web of internal tunnels throughout, which results in a loss of strength. This wood is then commonly injected with a consolidation agent to compensate for its internal weakness. The study is made to compare the strength of the most commonly used consolidating agents. These lection of agents was determined by interviewing conservators in Sweden, Norway and Denmark, as well as from accessible literature and research. In the study the selected consolidating agents are injected into trial pieces of artificially compromised wood which are in turn exposed to strength testing in a laboratory setting. The trial pieces are squares of birch wood measuring 45x45x45mm which have several 2mm holes drilled into them lengthwise, to simulate the beetle's pathways. Two types of strength testing were carried out to test the consolidation agents and their effect. One testmeasures the strength by compression across the grain, while the other puts pressure on either side, forcing the piece to fail along the grain. The results of the testing do not distinctively show that the consolidating agents impose any significant strength to the trial pieces. There is however a tendency of increased strength shown in the cross grain compression tests. To draw any firm conclusions, additional research is required.

L'objectif de cette étude est d'analyser la complémentarité des signaux olfactifs et acoustiques indispensable à la rencontre des sexes chez Nezara viridula (Heteroptera, Pentatomidae), et de caractériser l'importance éthologique de la variabilité inter-populations de ces signaux. Dans un premier temps, nous avons basé notre travail sur l'étude des différentes étapes constituant le SMRS. Ainsi nous avons mis en évidence le rôle central des chants d'appel émis par les femelles. Ces chants d'appel permettent aux mâles de localiser la femelle sur la plante, alors que la phéromone sexuelle mâle ne permet pas aux femelles de le localiser précisément. Cependant, des composés olfactifs émis par le mâle, sont responsables du déclenchement des chants d'appel chez les femelles. Le mâle répond aux chants d'appel en s'orientant vers la femelle, et en émettant des chants de cour de façon à synchroniser son chant avec celui de la femelle. Le mâle module également l'émission de sa phéromone sexuelle en fonction de la présence/absence de partenaire sexuelle potentielle. Dans un deuxième temps, nous nous sommes intéressés aux déterminismes comportementaux de la sélection du partenaire sexuel. Pour cela nous avons tout d'abord étudié la répétabilité des signaux phéromonaux et acoustiques. Nous avons ensuite mis en évidence la sélectivité des femelles par rapport aux composés olfactifs émis par les mâles. De même, les mâles répondent préférentiellement aux chants d'appel de leurs femelles, même s'ils reconnaissent et répondent aux chants d'appel émis par des femelles "étrangères". Nous discutons en quoi l'évolution de ces signaux peut être le résultat d'une sélection disruptive provenant d'une adaptation secondaire à des milieux différents. Nous intégrons l'ensemble de ces résultats afin de proposer des pistes dans la mise en place d'un programme de lutte intégrée permettant de contrôler ce ravageur
Male N. Viridula produce a volatile pheromone, attracting females. Natural male odour, and, less efficiently, a blend of the bisabolene epoxides and bisabolene, trigger calling behaviour of mature females. Thus, male pheromone is also active in reciprocal short-range communication through positive feed back. Once male and female on a same plant, they communicate with substrate borne vibrations. Male uses the vibratory signals emitted by female to locate her. Thus, female calling song (FCS) is an essential component of the SMRS. Male responds systematically to FCS by emitting male courtship song (MCrS) and modulates temporal and spectral characteristics of its song in synchronisation with female pulse train. Moreover, monitoring of pheromone production showed that males increase emission of the sex pheromone when stimulated with FCS, compared to control insects which showed a tendency to decrease their emission, and males stimulated with a male rivalry song, which showed a stable emission of pheromone. It has been repeatably observed that the song characteristics and the pheromone composition differ between geographically isolated populations. At this point of our study, we considered it was of importance to examine how far the responses of male and female bugs are specific, and allow some discrimination between potential mates. Females respond better to a natural odour of males from their own populations compared to another one. In the same way, mâle responds to FCS of females from their own population by emitting more MCRS and by increasing his amount of pheromone. In conclusion, there is no behaviour barrier between geographically isolated populations of N. Viridula. We discuss the variability of the recording signal to the adaptation of each population to different host plants. There results are discussed to propose a strategy of pest management

Cotton aphid, Aphis gossypii Glover, fecundity, nymph development and honeydew production were studied in the laboratory. Apterous adult females produced an average of 1.7 nymphs per day and the nymphs (four instars) developed to adults in an average of 4.1 days at 26.7° C in the laboratory. Average longevity of adults was 16.1 days. More honeydew drops were produced by one-day old nymphs than three- or four-day old nymphs. Numbers of honeydew drops produced on a day to day basis were highly variable and did not show a distinct pattern of production. More honeydew drops, sugars and progeny were produced by adults at 26.7° C compared with 15.6 or 32.2° C. Increasing times of exposure of clean cotton lint to aphids and the resulting increasing amounts of honeydew sugars under laboratory and field conditions were significantly related to increasing cotton lint stickiness as measured with a thermodetector.

The in vitro rate and cationic composition of the fluid secreted by the Malpighian tubules of the African migratory locust, Locusta migratoria migratoroides L, was investigated in this study. The concentrations of the elements Na, K, P, S, CI, Mg and Ca within Locusta Malpighian tubule type 1 cells, and the surrounding basement membrane, were quantified. Inhibitors and stimulators of fluid production were used to perturb the normal secretory state of the tubule cells. The rate of fluid secretion under control conditions was between 1.82 and 1.33. nl min(^-1) The fluid [K(^+)] was approximately 126mM, and [Na(^+)] 51 mM. The basement membrane was characterised by high [Na] and [CI] whilst a gradient of [K](_i) was observed. [K](_i) rose from approximately 193 mmol Kg(^-1) d.w. at the basal infoldings to 481 mmol Kg(^-1) d.w. at the apical microvillar border. The central cytoplasmic [K](_i) was 348 mmol Kg(^-1) d.w., estimated as 116mM. [Na](_i) and [Cl](_i) were generally lower, being 57mM and 29mM respectively in the central cytoplasm. Only K assumed a concentration gradient. Intracellular mass dense concretions were observed. Three types were present, the first rich in P and Ca, the second, rich in S, Na and K, and the third, rich in Mg, K and Na. The fluid production inhibitors furosemide (1mM) and bafilomycin A(_1) (1µM) raised the [N(^+)] in the secreted fluid, and altered [K](_i), [Na](_i) and [CI](_i). Furosemide lowered [K](_i) but increased [Na](_i) and [Cl](_i). Bafilomycin lowered [K(^+)] in the secreted fluid, though [K](_i) increased. Both inhibitors abolished the [K](_i) gradient. Replacing K(^+) with Rb(^+) in the bathing saline slowed fluid secretion and lowered [K](_i) and [Cl](_i), though a gradient of [K](_i) was retained. Rb adopted an intracellular gradient which mirrored that of K. Rate of secretion data suggests Rb enters the cell basolaterally primarily via the Na(^+)-K(^+)-ATPase. The fluid secretion stimulator cAMP (1mM) lowered [K](_i), and raised [Na](_i) and [Cl](_i), but corpora cardiaca extract left these elements' concentrations largely unchanged. Stimulation with both corpora cardiaca extract and cAMP maintained the [K](_i) gradient. These stimulators changed the content and number of mass dense concretions present, in manner which suggested that these structures were important in ion transport. These findings support the current model of ionic transport in these cells, including the basolateral presence of an Na(^+)-K(^+)-2Cr cotransporter, and an apical proton-pumping V-type ATPase / K(^+)/nH(^+) antiport complex.

A potential strategy to produce safer and broadly protective influenza vaccines is to co-express, in the same cell host, multiple hemagglutinins (HA) with a matrix protein (M1) which self-assemble in virus-like particles (VLPs). This study demonstrates the suitability of combining stable expression and the baculovirus-expression vector system (BEVs) in insect Hi5 cells for production of such multi-HA Influenza VLPs. Stable pools of Hi5 cells expressing two HAs were generated and later infected with a M1-encoding baculovirus at two cell concentrations (CCIs; 2×106 cells/mL and 3×106 cells/mL). The HA concentration in culture supernatant was followed over time, with more productive infections observed at higher CCIs. To extend the culture time, a re-feed strategy was implemented based on the identification of key nutrients which were exhausted during cell growth. Afterwards, supplemented cultures infected at a CCI of 4×106 cells/mL allowed a 4-fold increase in HA concentration, at harvest, when compared to cultures infected at a CCI of 2×106 cells/mL. The production of multi-HA influenza VLPs using the aforementioned strategy could be successfully scaled-up to 2L bioreactor cultures with even higher volumetric (1.5-fold) HA yields. To surpass the unpredictability of gene expression promoted by the random integration strategy mentioned above, the recombinase-mediated cassette exchange (RMCE) technology was explored. The feasibility of having two cassettes flanked by distinct pairs of flippase recognition target sites (FRTs) was evaluated. Unfortunately, significant cross-interaction was observed between the selected pairs. To circumvent this bottleneck, a backup strategy consisting in the co-expression of two genes from the same locus after implementation of one cassette system, in insect Sf9 cells, was attempted. However, the isolated clones showed low expression of both M1 and HA proteins. Ongoing work focuses on the isolation of clones tagged in high expression loci by fluorescence activated cell sorter technology. This work demonstrates how the versatility of insect cell expression technology can be explored to produce Influenza VLPs as vaccine candidates.
A co-expressão de várias hemaglutininas (HA) e proteína da matriz (M1), no mesmo hospedeiro, formando partículas semelhantes a vírus (VLPs), constitui uma importante estratégia para desenvolver vacinas contra o vírus da gripe. Este trabalho mostra a combinação de uma linha celular estável de células de insecto com o sistema de expressão mediada por baculovírus para a produção deste tipo de VLPs. Foram estabelecidas duas populações de células de insecto Hi5, expressando duas HAs, posteriormente infectadas com um baculovírus contendo a proteína M1, a duas concentrações celulares diferentes (CCI; 2 e 3×106 cells/mL) sendo que a mais elevada demostrou ser a mais produtiva. De seguida, implementou-se uma estratégia baseada na adição de nutrientes específicos para prolongar o tempo de cultura. As culturas previamente suplementadas e infectadas a uma CCI de 4×106 células/mL produziram 4x mais HA comparativamente às culturas infectadas a uma CCI de 2×106 células/mL, não suplementadas. Esta estratégia foi também aplicada num biorreactor de 2L permitindo 1,5x mais produção, volumétrica, de HA comparativamente a experiências em pequena escala. De forma a ultrapassar a imprevisibilidade de uma integração aleatória, foi explorado o sistema de troca de cassete mediado por recombinase (RMCE). A viabilidade de um sistema com duas cassetes integradas flanqueadas por diferentes locais de reconhecimento (FRTs) foi avaliada, tendo sido observada a interação entre ambos os pares selecionados. Como segunda estratégia, foi implementado um sistema com uma cassete para co-expressão de dois genes em simultâneo, em células de insecto Sf9. Porém, os clones isolados mostram fraca expressão de M1 e HA, pelo que uma estratégia de isolamento de clones com expressão génica mais forte está em desenvolvimento utilizando uma tecnologia de sorteamento. Assim, este trabalho demonstra a versatilidade da tecnologia aplicada em células de insecto, que pode ser explorada para produzir VLPs multivalentes, com potencial para se tornar a próxima geração de vacinas para o vírus da gripe.

Dans le contexte de pénurie des ressources, croissance démographique, dégradation de l’environnement, la production d’aliments riches en protéines (en particulier) pour l’homme et les animaux devrait augmenter pour répondre à la demande. De nouvelles ressources sont actuellement explorées : légumineuses, algues, insectes... Ces derniers représentent une source de protéines plus durable comparée aux sources conventionnelles. Bien qu’ils soient consommés par de nombreuses populations en Asie, en Afrique et en Amérique du Sud, ce n’est pas le cas en Europe. Ainsi, pour faciliter leur utilisation dans l’alimentation européenne, les insectes peuvent être transformés en ingrédients afin de les incorporer dans des formulations d’aliments. Cependant, il existe très peu de données sur les méthodes de transformation à l’échelle pilote ou industrielle et sur l’impact du procédé sur la qualité des produits finis. Ce travail avait pour objectifs de mettre au point à l’échelle pilote un procédé de production de farine d’insecte pour l’alimentation, de caractériser cette farine et les autres produits obtenus, de caractériser les propriétés de la fraction protéique soluble et d’étudier l’impact du procédé sur les propriétés de celle-ci. Le Tenebrio molitor, candidat à l’élevage industriel, a été sélectionné pour cette étude. Un procédé thermomécanique de production de farine a été mis au point à l’échelle pilote. Il a permis la production d’une farine d’insecte riche en protéines de 72% (bs) avec 14% (bs) de lipides et 4% (bh) d’eau. Le profil en acides aminés des protéines de cette farine répond aux besoins de la nutrition animale et de l’alimentation humaine avec une bonne efficacité protéique (estimée à 2,5). Le rendement de la production de 20% (bh) (64% bs) est semblable à celui de la production de farine de poisson (20% bh). Parallèlement, l’huile d’insecte, principale coproduit a également été produite. Elle est riche en acides palmitiques, et en acides gras essentiels ω9 et ω6. Elle peut être utilisée en alimentation ou dans d’autres domaines. Bien que le procédé ait un impact sur les propriétés physicochimiques des protéines solubles après la transformation des larves en farine, les fractions protéiques solubles de la farine et des larves ont les mêmes propriétés moussantes et émulsifiantes semblables à celles du lait et de ASB à 4 et 2% respectivement. Les protéines de la farine ou des larves peuvent être également utilisées pour leurs propriétés fonctionnelles. Ce travail contribue à la compréhension des protéines d’insectes et à l’extrapolation industrielle dans une perspective de conception de la bioraffinerie
In the context of resource scarcity, population growth, environmental degradation, and food supplies dependency, the production of protein-rich feed and food should increase in order to meet the demand. New resources are currently being explored as vegetal, algae, insects... This last one is environmentally friendly and represents a more sustainable protein source as compared to conventional livestock farming. Although insects are consumed by a lot of people in Asia, Africa and South-America, this is not the case in Europe. In order to meet European consumers' preferences, they need to be processed or transformed into ingredients to become a part of formulation products (i.e. powders). However, very little knowledge regarding processing methods at a pilot or industrial scale, and the composition and impact of process on the properties of insect-based ingredient exists and is available. The aim of this work was to design a process for the production of meal rich in proteins from insect at a pilot scale, to characterize it for feed and food applications, to characterize the properties of its soluble proteins, and to study the impact of the process on these properties. The Tenebrio molitor, candidate for rearing at an industrial scale, was chosen in the frame of this study. A thermo-mechanical process was designed at a pilot scale. It allowed the production of a protein-rich insect meal of 72% (bs) with 14% (bs) of lipids and 4% (bh) of water. The amino acid profile of this meal proteins meets the needs of animal nutrition and human nutrition with good protein efficiency (estimated at 2.5). The production yield of 20% (bh) (64% bs) is similar to that of fishmeal production (20% bh). In parallel, insect oil (co-product) were also produced. It is rich in acid palmitic, and essential fatty acids ω9 and ω6. It can be used in feed, food, cosmetic or bioenergy. Although the process has an impact on the physicochemical properties of soluble proteins after the transformation of larvae into flour, the soluble protein fractions of flour and of larvae have the same foaming and emulsifying properties similar to those of milk and BSA at 4 and 2% respectively. The meal proteins could be used in feed and food for their functional properties. This work contributes to insect protein understanding and the industrial extrapolation in a perspective of biorefinery designing

The synthesis of p-menthane-3,8-diol via the acid-catalyzed cyclization of citronellal in a dilute aqueous sulphuric acid medium was investigated using conventional batch and continuous systems in order to develop a commercial production process for said p-menthane-3,8-diol (PMD). The batch studies conducted during the first part of this study showed that the formation of PMD from citronellal occurs via an intra-molecular Prins reaction that results in the formation of both the desired PMD product, as well as the partially hydrated isopulegol. It was shown that the formationof the by- product, PMD-acetal, results from the reaction between an intermediate, 5-methyl-2- isopropylcyclohexanol, and the citronellal starting material, and not from the reaction between PMD and citronellal as previously reported. Kinetic studies confirmed the existence of a complicated kinetic model. The formation of PMD from citronellal displayed typical pseudo first order kinetics up to conversions of 70 after which the kinetic model becomes complicated as the result of the establishment of quasi equilibrium reactions between PMD and isopulegol (dehydration of PMD and hydration of isopulegol) and between PMD the PMD-acetal, both systems being acid catalysed. The PMD-acetal formation reaction appears to be second order with respect to PMD. Scale-up studies of the batch process to 30L and 50L scales showed that it would be extremely difficult to limit the level of PMD-acetal formation below the desired level of 1 percent, even if citronellal conversions are restricted to about 50 percent. During studies conducted on a commercially availablemicro-structured organic synthesis plant (OSP) it was shown that it is possible to perform the PMD reaction as a continuous process. The results obtained showed that the use of a micro-mixer such as the caterpillar micro-mixer did not provide enough residence time in order for desirable conversions (- 40 percent) to be obtained. By combining themicro-mixer with delay-loops of different thicknesses and lengths, and using increasing reaction temperatures, it was shown that the conversion of citronellal could be improved to some extent, but compared poorly to the expected conversions for a well-stirred batch reactor. By packing selected delay loops with inert SiC particles, improved mass transfer was observed between the organic and aqueous phases as reflected in the increased conversion of citronellal. Using the observations that were made during the use of the OSP, a continuous-flow, tubular reactor system was designed and constructed. Advanced statistical techniques were used to investigate the effect of variables such as temperature, acid concentration, reactor length, flow rate and the organic to aqueous ratio on the rate and selectivity of the reaction. Mathematical models were derived for citronellal conversion, yield of PMD and yield of PMD- acetals, and used to predict the concentrations of citronellal, PMD and PMD-acetals at set experimental conditions. The results obtained showed that it was possible to obtain a product which approached desired specifications.Downstream processing of the PMD reaction mixture as it exits the reactor requires phase separation and neutralization of the acid catalyst solution, followed by further work-up to recover unreacted starting material and intermediates for recycle back to the synthesis reactor, followed by purification of crude PMD to the desired specification. The study showed that neutralization, prior or after phase separation, does not affect the selectivity of the PMD to such a great extent, but does influence the relative conversion due to extended contact of the catalyst with the organic phase after the reaction is terminated. Recovery of unreacted citronellal and isopulegol could be achieved by a simple vacuum evaporation step, which may either be carried out in a batch manner using traditional distillation equipment, or in a continuous process using wiped-film (short path) techniques. It was also shown that selective crystallization of PMD from the crude product mixture by addition of a solvent, such as heptanes or hexane proved to be the best way of achieving the desired product specification.

We determined honeydew production by male and female sweetpotato whiteflies and the effects of temperature on honeydew production of each sex. We also determined honeydew production by each nymphal instar. Overall, adult SPW produced more honeydew than nymphs. Adult females produced more honeydew than males. The relative differences between honeydew production for males and females and between amounts adults produced compared with nymphs were consistent. However, honeydew production by adult and nymph individuals was subject to large degrees of variation.

Soybean, Glycine max (L.) Merr., is planted across a vast amount of land in the Mid-Southern U.S. (Arkansas, Louisiana, Mississippi, and Tennessee), and yield responses to defoliation can vary. Experiments were conducted during 2015-2017 evaluating how soybean yield responds to multiple and continuous defoliation, as well as planting date and plant population. Multiple defoliation events were evaluated by defoliating soybean at varying levels at V3, V6, and both growth stages. There was no interaction between defoliation occurring at V3 and V6 growth stages, indicating that the impact of each defoliation event was independent of the other. The effect of continuous defoliation was evaluated by defoliating soybean weekly, beginning at V2. Defoliation continued throughout the vegetative growth stages or throughout the entire growing season, and was compared to the same defoliation level occurring one time at R3. Continuous defoliation during vegetative growth stages only, did not reduce yield at any of the levels tested. Defoliation occurring throughout the growing season reduced yields more than a one-time defoliation event at R3, but only when defoliation levels exceeded the 20% defoliation threshold. This indicates that thresholds do not need to be modified to account for multiple or continuous defoliation. To evaluate the effect of planting date on yield loss from defoliation, soybean was planted at six planting dates beginning in early-April and continuing through mid-June. Each planting date included a defoliated treatment and an undefoliated control. It was determined that later planted soybean lose a greater amount of yield than earlier planted. Higher yielding soybean also lost more yield than lower yielding soybean at every planting date until Mid-June. It was concluded that late planted soybeans could benefit from a lower treatment threshold. The effect of plant population on yield loss from defoliation was evaluated by planting soybean at five populations ranging from 123,500 seeds/ha to 420,070 seeds/ha. A undefoliated control and a defoliated treatment was included for each plant population. Defoliation significantly reduced yields only where final plant populations were lower than 192,800 plants/ha. This indicates that fields with substandard plant populations are more susceptible to yield loss from defoliating pests.

Résumé Une demande croissante sans cesse de produits alimentaires à base d’animaux influence la productivité des systèmes mondiaux de production alimentaire, et des mesures indispensables pour freiner la dégradation de l’environnement promettent des effets similaires. Si les scénarios de demande future peuvent être satisfaits de manière durable, cela dépend notamment de la possibilité de réduire de manière significative l'impact de l'aquaculture et de l'élevage sur l'environnement. Des recherches récentes suggèrent que l'utilisation d'aliments à base d'insectes (IBF) pourrait apporter une contribution significative à cet égard, et des arguments valables sont avancés pour soutenir cette hypothèse. Les larves de mouches, comme celles des mouches domestiques (Musca domestica) ou des mouches soldat noir (Hermetia illucens), sont en mesure de puiser des nutriments dans un large éventail de ressources organiques, y compris celles impropres à la consommation humaine. Cela crée la possibilité de convertir (et de réduire considérablement) les déchets organiques de faible valeur, tels que le fumier ou le sang animal, en protéines de haute qualité et en énergie alimentaire, qui se sont avérés appropriés pour nourrir différents poissons d'aquaculture et du bétail monogastrique.Bien que le concept IBF promet d’importants avantages et ait démontré sa faisabilité technique, il n’existe encore aucun système établi permettant de tester les avantages supposés en termes de durabilité. Dans cette thèse, nous avons essayé de surmonter cette lacune par la modélisation de tels systèmes. Notre objectif central était d'identifier les aspects influençant le potentiel d'application des IBF dans différents contextes géographiques et de définir des voies d'optimisation pour une mise en œuvre durable. En nous basant sur des données expérimentales recueillies lors d'essais d'élevage menés en Europe (Espagne et Slovaquie) et en Afrique de l'Ouest (Ghana et Mali), nous avons formulé la conception d'un ensemble de versions de systèmes améliorés élevant M. domestica et H. illucens sur différents substrats organiques de faible valeur. Les modèles de production génériques ont servi comme base d’une analyse du cycle de vie ex ante, dans laquelle nous avons exploré les performances des systèmes à l’aide d’analyse du cycle de vie environnementale (ACV) et de l’analyse des coûts du cycle de vie (ACCV).Les ACVs et ACCVs ont montré que les performances environnementales et économiques des IBF sont largement fonction de l’efficacité de conversion des systèmes, de l’organisation du processus de production (c’est-à-dire de l’apport de main-d’œuvre et d’équipements technologiques) et du contexte géographique. La combinaison de ces facteurs a fourni des avantages pour les configurations simplistes utilisées dans la production de M. domestica en Afrique occidentale tropicale dans des conditions de ponte naturelle (c'est-à-dire d'inoculation de substrat par le biais de mouches présentes à l'état naturel). L'inoculation artificielle (c'est-à-dire l'inoculation du substrat par le biais de larves nourries provenant d'une colonie d'adultes en captivité), utilisée dans la production de H. illucens en Afrique de l'Ouest et de M. domestica dans le sud de l'Espagne, a favorisé une efficacité de conversion élevée, mais a augmenté les impacts environnementaux et les coûts, parce que le système complexe et l'organisation de processus à forte intensité de main-d'œuvre ont considérablement accru les intrants de main-d'œuvre et d'infrastructures de production.Une comparaison avec des aliments conventionnels riches en protéines a mis en évidence des inconvénients environnementaux et économiques pour les modèles de production actuels des IBF, notamment en ce qui concerne les aliments végétaux (par exemple, le tourteau de soja). Les disparités entre les alimentations IBF et conventionnelles reflètent l’utilisation des capacités sub-optimaux des systèmes (effet d’économie d’échelle insuffisant), ainsi que la perte d’énergie et de biomasse le long de la chaîne trophique (producteurs autotrophes vs consommateurs hétérotrophes). Les résultats soulèvent des doutes légitimes sur les avantages en termes de durabilité d’une mise en œuvre d'insectes dans les chaînes de valeur agricoles actuelles. Le succès commercial dépend en grande partie du niveau de salaire spécifique au contexte, des prix des substrats d'élevage et de la manière dont les marchés évaluent les multiples fonctions que les insectes sont capables de fournir. S'agissant de la performance environnementale, nos résultats nous amènent à conclure que la production d'IBF ne présente aucun avantage par rapport aux aliments conventionnels.L’évaluation de systèmes de production encore hypothétiques impliquait une bonne quantité d’hypothèses et d’approximations. Étant donné ces multiples sources d'incertitude et compte tenu du fait que seul un nombre limité de conceptions de systèmes possibles sont prises en compte, les déclarations sur le potentiel d'application d'IBF n'ont aucune validité universelle et doivent être interprétées avec prudence. Cependant, nos résultats contribuent à une meilleure compréhension des facteurs influant sur le potentiel d’application des systèmes de production d’insectes et constituent un point de référence précieux pour les discussions scientifiques et les activités de recherche et développement futures visant à mettre en place des modes de production alimentaire durables.Bien que nos recherches n’apportent aucun soutien aux avantages environnementaux ou économiques supposés de l’utilisation d’insectes dans l’alimentation animale, il est possible que leur utilisation comme aliment destiné à la consommation humaine directe (c’est-à-dire comme substitut possible aux produits à base de poisson et de viande) constitue une solution durable aux problèmes actuels et futurs. Nous recommandons donc aux recherches futures de se concentrer sur les techniques permettant d'exploiter les insectes comme nourriture.
Doctorat en Sciences
There are a few details that I could not specify in the available input fields. I would like to ask you kindly to add the following information: (1) Prof. Erik Mathijs (KUL) is my second co-supervisor; (2) Next to the Jury members listed, there are Prof. Matthias Finkbeiner (TU Berlin) and Prof. Theo Niewold (KU Leuven), which I could not enter manually.
info:eu-repo/semantics/nonPublished

El principal objectiu fou produir sistemes d’encapsulació basats en emulsions emprant ingredients i tecnologies sostenibles. S’han combinat amb èxit la concentració per osmosi directa i la emulsificació per membranes en la valorització d’extracte de polifenol de garrofa. Especies d’insectes amb potencial per ser comercialitzades s’han valorat com a emulsificants. Fraccions proteiques de larva de mosca de soldat negre (BSFPC, Hermetia illucens) i d’escarabat búfal (LMPC, Alphitobius diaperinus) s’han utilitzat per estabilitzar emulsions simples i múltiples mitjançant emulsificació amb membranes dinàmiques de porositat controlada. BSFPC presenta millors resultats que la proteïna de sèrum de llet (WPI) per l’estabilització d’emulsions d’oli de llimona amb una fracció d’oli del 40%. Per emulsions d’oli de gira-sol, BSFCP es comporta de forma similar a WPI. En emulsions W1/O/W2, LMPC es comporta de manera similar a WPI i a la proteïna de pèsol (PPI) durant cicles de congelació-descongelació, en medi àcid i bàsic i durant l’emmagatzematge a diferents condicions. A més, mostra millors propietats que WPI i PPI per estabilitzar emulsions a 90ºC, indicant els avantatges d’utilitzar-les en aliments que han de passar per tractaments tèrmics. En canvi, no poden igualar l’estabilitat de les emulsions amb WPI quan es varia el gradient de pressió osmòtica entre les dues fases aquoses. S’ha comprovat l’eficàcia de les proteïnes d’insecte per encapsular un polifenol comercial en sistemes sòlids a partir d’emulsions W1/O/W2 assecades per atomització o liofilització. En el procés de fraccionament de proteïnes s’ha comprovat que solvents verds, com l’etanol, isopropanol i 2-metiltetrahidrofurà, es poden utilitzar alternativament a l’hexà en el desgreixat de molturats d’insecte. El rendiment d’extracció i les propietats emulsificants de les proteïnes resultants confirmen l’eficàcia d’aquests solvents. El resultats demostren que els concentrats de proteïna d’insectes utilitzats són una bona alternativa a les proteïnes làctiques o de plantes per la producció de sistemes d’encapsulació basats en emulsions.
El principal objetivo fue producir sistemas de encapsulación basados en emulsiones utilizando ingredientes y tecnologías sostenibles. Se han combinado con éxito la concentración por ósmosis directa y la emulsificación por membranas en la valorización de extracto de polifenol de algarroba. Especies de insectos con potencial para ser comercializadas se han valorado como emulsificantes. Fracciones proteicas de larva de mosca de soldado negro (BSFPC, Hermetia illucens) y de escarabajo búfalo (LMPC, Alphitobius diaperinus) se han utilizado para estabilizar emulsiones simples y múltiples mediante emulsificación con membranas dinámicas de porosidad controlada. BSFPC presenta mejores resultados que la proteína de suero de leche (WPI) para la estabilización de emulsiones de aceite de limón con una fracción de aceite del 40%. En emulsiones de aceite de girasol, BSFCP se comporta de forma similar a WPI. En emulsiones W1/O/W2, LMPC actúa como WPI y la proteína de guisante (PPI) durante los ciclos de congelación-descongelación, en medio ácido y básico y durante el almacenamiento en diferentes condiciones. Además, muestra mejores propiedades que WPI y PPI para estabilizar emulsiones a 90ºC, indicando la ventaja de utilizarlas en alimentos que tienen que pasar por tratamientos térmicos. En cambio, no pueden igualar la estabilidad de las emulsiones con WPI cuando se varía el gradiente de presión osmótica entre las dos fases acuosas. Se ha comprobado la eficacia de las proteínas de insecto para encapsular un polifenol comercial en sistemas sólidos a partir de emulsiones W1/O/W2 secadas por atomización o liofilización. En el proceso de fraccionamiento de proteínas se ha comprobado que solventes verdes, como el etanol, isopropanol y 2-metiltetrahidrofurano, se pueden utilizar alternativamente al hexano en el desengrasado de molturados de insecto. Los resultados demuestran que los concentrados de proteína de insectos utilizados son una buena alternativa a las proteínas lácteas o de plantas para la producción de sistemas de encapsulación basados en emulsiones.
The focus of the thesis is to produce emulsion-based encapsulation systems using both sustainable ingredients and technologies. Specifically, valorisation of an agri-food by product (carob pulp polyphenol) was proved feasible coupling forward osmosis for concentration and membrane emulsification for encapsulation. Insect powder defatting by solvent extraction was investigated using green solvents (ethanol, isopropanol, and 2-methyltetrahydrofuran), paying special attention to defatting yield and the techno-functional properties of the resulting protein fractions, particularly emulsifying ability. Single and double emulsions stabilised with sustainable protein sources from insects, black soldier fly (Hermetia illucens) and lesser mealworm (Alphitobius diaperinus) larvae, have been successfully produced for the first time by a low-energy high-throughput emulsification technology based on dynamic membranes of tunable pore size (DMTS). H. illucens protein concentrate showed superior ability to stabilize higher lemon oil fraction (40 wt%) compared to whey protein isolate. A commercial polyphenol extract was encapsulated in W1/O/W2 emulsions stabilised with A. diaperinus protein concentrate. These emulsions displayed a comparable stability under freeze-thaw cycles, storage conditions, acidic, and alkaline conditions than the ones stabilized with WPI, and better than the ones stabilized with pea protein. However, they were less able to withstand osmotic pressure differences compared to whey protein. Insect protein stabilized W1/O/W2 emulsions showed less changes in droplet size distribution at the highest temperature tested (90ºC) than the ones stabilized with whey or pea protein, pointing out the benefit of using insect proteins in emulsions that need to undergo heat treatment. Solid microcapsules were successfully produced from refined polyphenol loaded W1/O/W2 emulsions stabilized with insect protein by spray drying or freeze drying. The results demonstrate the insect protein concentrates assessed are a promising sustainable ingredient to replace diary and plant proteins in the emulsion-based encapsulation systems.

The need for automation is increasing at Sandvik Coromant. However, high equipment-, education-, and implementation costs connected to automation are usually obstacles when automating smaller and simpler tasks. During the recent years, a new type of robot has been introduced to the market. It is called a collaborative robot, or cobot. It is easy to use/program, has a lower cost and has built-in safety features. The user friendliness of the robot as well as the built-in safety features could lower the implementation costs of these robots to open new areas for automation. This thesis evaluates a cobot from Universal Robots, UR5, to determine if such technology could be useful at Sandvik Coromant’s Insert Plant in Gimo, Sweden. This would be done by performing a physical demonstration of a UR5 put into Sandvik Coromants production. The task includes identifying, conceptualizing and evaluating the possible applications where cobots could create the most value. The work showed that cobots are indeed easy to work with and could create more areas of automation. However, the way Sandvik Coromant looks at automation should change for the technology to make the biggest impact. A company guideline must be setup regarding the use of the built-in safety features to set a limit of what is acceptable when creating an open cell environment at the factory. The results and conclusion of this thesis will hopefully lead to the implementation of collaborative robots at Sandvik Coromant which will result to lower automation costs, better working environment and higher production.
Automationsbehovet på Sandvik Coromant har ökat under de senaste åren. Höga utrustnings-, utbildnings-, och utvecklingskostnader kopplade till automation sätter oftast stopp för automation av mindre och enklare uppgifter. Under de senaste åren har en ny typ av robot blivit introducerad på marknaden. Den kallas för samverkansrobot, eller cobot. Den är enkel att använda/programmera, har lägre kostnad och inbyggda säkerhetsfunktioner. Användarvänligheten av roboten samt de inbyggda säkerhetsfunktionerna kan leda till lägre implementationskostnader, som öppnar nya möjligheter för automation. Detta mastersarbete utvärderar en cobot från Universal Robots, UR5, för att fastställa om sådan teknologi kan vara användbar på Sandvik Coromants skärfabrik i Gimo, Sverige. Utvärderingen kan göras genom att fysisk demonstration av UR5 i Sandvik Coromants produktion genomförs. Uppgiften består av att identifiera, konceptualisera och utvärdera de applikationer i fabriken där samarbetsrobotar skulle kunna skapa mest värde. Arbetet visade att cobots är enkla att arbeta med och kan skapa mer möjligheter för automation. Dock så måste sättet Sandvik Coromant ser på automation att ändras för att teknologin ska ha störst inverkan. En intern riktlinje måste skapas angående användandet av de inbyggda säkerhetssystemen för att sätta en gräns på vad som är accepterat ur en arbetsmiljösynpunkt när det gäller öppna robotceller. Resultaten och slutsatsen av arbetet kommer förhoppningsvis leda till implementering av samverkansrobotar på Sandvik Coromant som i sin tur leder till lägre automationskostnader, bättre arbetsmiljö och högre produktion.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
O presente trabalho teve por objetivos buscar alternativas às tradicionais sacolas de papel manteiga, buscando proteção segura aos ataques de insetos-praga e pássaros, maior facilidade no momento da colheita de pêssegos, e verificando também o custo e a praticidade das embalagens. O trabalho foi realizado no Sítio Santa Maria, no município de São Luis do Paraitinga, SP, durante o período de agosto a novembro de 2006, em um pomar comercial de pêssego do cultivar Aurora 2 , de três anos de idade, conduzido em sistema de vaso moderno e espaçamento de 6 x 4 m. O delineamento experimental utilizado foi em blocos casualizados, onde cada planta foi considerada um bloco, utilizando-se 15 tratamentos, 8 repetições (blocos) e 10 frutos por tratamento. Os tratamentos foram os seguintes: T1 - Sacolas de TNT branco de 45 Gramaturas por m2 (45G/m2) fechado, T2 - Sacolas de TNT branco de 45 Gramaturas por m2 (45G/m2) aberto, T3 - Sacolas de TNT branco de 20 Gramaturas por m2 (20G/m2) fechado; T4 - Sacolas de TNT branco de 20 Gramaturas por m2 (20G/m2) aberto; T5 - Sacolas de polipropileno microperfurado transparente (furos de 1mm) fechado; T6 - Sacolas de polipropileno microperfurado transparente (furos de 1mm) aberto; T7 - Sacolas de polipropileno microperfurado 2 transparente (furos de 2mm) fechado; T8 - Sacolas de polipropileno microperfurado transparente (furos de 2mm) aberto; T9 - Sacolas de polietileno microperfurado leitoso (furos de 1mm) fechado; T10 - Sacolas de polietileno microperfurado leitoso (furos de 1mm) aberto; T11 - Sacolas de polietileno microperfurado leitoso (furos de 2mm) fechado; T12 - Sacolas de polietileno microperfurado leitoso (furos de 2mm) aberto; T13 - Sacolas de papel impermeável fechado; T14 - Sacolas de papel impermeável aberto; T15 - Testemunha (sem ensacamento). Os resultados mostraram que o ensacamento não influenciou...
This research aimed to look for alternative bags that could offer protection against fruit-flies and oriental moth attack, offer easy procedure at the harvesting time of peach crops and verify two aspects referred to the bags: the practicability and the cost. The work was conducted at Santa Maria Farm in São Luis do Paraitinga County (São Paulo State), from August to November of 2006, in a tree-year-old peach crop. The cultivar Aurora 2 was conducted in modern vase system with 6m between rows and 4 m between plants. The experimental design followed randomized block, where each plant was considered one block. There were 15 treatments, 8 replications (blocks) and 10 fruits per treatment. The treatments were: T1 - white and closed TNT (tissue non-tissue) bag (45g/m2), T2 - white and opened TNT bag (45g/m2), T3 - white and closed TNT bag (20g/m2), T4 - white and opened TNT bag (20g/m2), T5 - transparent polypropelene microperfurated and closed bag (1mm of diameter), T6 - transparent polypropelene microperfurated and opened bag (1mm of diameter), T7 - transparent polypropelene microperfurated and closed bags (2mm of diameter), T8 - transparent polypropelene microperfurated and opened bag (2mm of diameter), T9 - milky microperfurated polyetilene and closed bag (1mm of diameter), T10 - milky microperfurated polyetilene and opened bag (1mm of diameter), T11 - milky microperfurated polyetilene and closed bag (2mm of diameter), T12 - milky microperfurated polyetilene and opened bags (2mm of diameter), T13 - closed and waterproof paper bag, T14 opened and waterproof paper bag, T15 - check treatment (unbagging fruits). The results showed that bagging did not influenced chemical fruits characteristics, such as, pH, soluble solid contents (SS), titratable acidity (TA), SS/TA ratio, however, produced reasonable bigger and havier fruits. The fruit epiderm color was influenced by baging... (Complete abstract click electronic access below)

Les interactions interindividuelles permettent la transmission de l’information, la dispersion des pathogènes et la mise en place des comportements dans une population. Cette thèse a permis d’évaluer l’influence des interactions sociales sur le succès de fondation colonial des différents reproducteurs de deux termites européens, l’invasif Reticulitermes flavipes et le natif R. grassei. Les résultats révèlent (i) un meilleur succès de fondation des reproducteurs primaires de R. flavipes, (ii) une organisation biparentale des soins aux jeunes toutes espèces confondues et (iii) une communication et des soins aux oeufs propres aux caractères invasif et natif des espèces d’étude. Pour finir, (iv) une meilleure survie et communication a été observée dans les colonies fondées avec reproducteurs secondaires tandis (v) qu’une communication supérieure et une survie moindre sont observées pour R. flavipes. Les origines évolutives de l’organisation biparentale et des variations de succès de fondations sont discutées
Individual interactions permit information transmission, pathogen dispersion and shape behavioral strategies in a population. This thesis has permit to explore the influence of social interactions on the colonial foundation success of two European termites, the invasive Reticulitermes flavipes and the native R. grassei. The overall results revealed (i) a better foundation success of primary reproductives of R. flavipes, (ii) a biparental organisation of parental care in both species (iii) a level of communication and egg care reflecting native and invasive status of the two species studied. To finish, (iv) better survival and communication rates were observed in colonies founded with secondary reproductives than in colonies without any and (v) a better communication rate and a weaker survival rate for R. flavipes foundations with or without secondary reproductives. Evolutive origins of biparental care and of the variations of foundation success observed are discussed

Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2004.
Includes bibliographical references (p. 119-120).
Three Dimensional Printing (3DP) is a solid freeform fabrication process used to generate solid parts directly from three-dimensional computer models. A part geometry is created by selectively depositing binder into sequentially spread layers of powder. In slurry-based 3DP, a suspension of powder in a solvent is used to form the powderbed layer. This slurry-based powderbed yields higher green density and part resolution than dry powder-based 3DP because of smaller particle size. Vector printing requires that the printhead trace and define the external geometries of a part before raster filling the interior, a new approach in comparison to conventional, raster-only printing. Drop-on-demand (DOD) printheads allow binder droplets to be ejected when needed rather than relying upon charge-and-deflect mechanisms used in continuous jet printheads. Integrating these concepts for vector, DOD printing has the potential to enhance the 3DP process by providing greater part resolution and surface finish. The 3DP slurry-based process and vector, drop-on-demand printing are examined as potential methods to produce Tungsten Carbide-Cobalt (WC-Co) tooling inserts. The research focuses on three fundamental process steps: (1) development of a stable slurry, (2) determination of jetting parameter values for optimal powderbed deposition, and (3) implementation of vector, DOD printing for the binder. Due to unforeseen circumstances, the first two objectives are only briefly introduced in Chapter 1 and summarized in Chapter 3. Further details may be found in the Diplomarbeit document of Olaf Dambon. Two approaches are explored to develop a stable, jettable slurry. One method involves using a water-based Tungsten Carbide slurry and a
(cont.) Cobalt Acetate binder; the other method utilizes an alcohol-based Tungsten Carbide-Cobalt slurry and an organic binder. Various suspension properties, such as sedimentation density and viscosity, are measured to assess the degree of slurry stability. After adequate slurry formulations are developed, an investigation of powderbed formation is conducted. Due to the low solubility limit of the Cobalt salt in water and the persistent defects in water-based slurry powderbeds, the alcohol-based approach is pursued and, because of its greater efficacy, is used for optimizing powderbed jetting parameters. An effective combination of line spacing, flow rate, and drying time is determined for producing powderbeds with minimal surface roughness and high packing density. Experiments are subsequently conducted in vector DOD printing of various geometries using a piezo-actuated, drop-on-demand printhead and Bridgeport three-axis milling machine. A Hewlett-Packard inkjet cartridge is initially used for vector testing of the milling machine; a Siemens PT-88S printhead is used to assess and optimize binder droplet formation parameters, such as voltage waveform and fluid properties. Functional conditions for vector printing and DOD droplet generation are developed and deliver acceptable performance. Successfully printed geometries with high-definition lines (140-170 [mu]m line width) and smooth surface finish are produced using sanded, jetted alumina slurry powderbeds. Following necessary refinements in slurry redispersion and slurry-binder compatibility, the same vector process can be repeated with jetted WC-Co slurry powderbeds.
by David Guo.
S.M.

Cotton lint contamination from honeydew excreted by sweetpotato whiteflies, Bemisia tabaci (Gennadius), is a serious problem in the textile industry resulting in reduced lint processing efficiency. The insect growth regulators, Applaud® and Knack®, provide effective control of sweetpotato whiteflies on cotton by interfering with their reproduction and development. Protection from honeydew lint contamination is attributed to reduced sweetpotato whitefly populations. We investigated the potential direct effect of Applaud and Knack on sweetpotato whitefly honeydew production. In the field, amounts of the major sugar components of honeydew produced by adults and nymphs collected on day six following Applaud or Knack applications to cotton field plots were not significantly different compared to amounts produced by those collected from untreated plots. In the laboratory, adult mortality and amounts of honeydew sugars produced by adults were not affected by confinement for 48 h on Applaud or Knack residues from cotton leaf dips or following nebulizer contact spray applications. In contrast, mortality of first and second instar nymphs on leaves was higher on day six following leaf dips in Applaud solutions compared with leaf dips in Knack or water solutions. Nymph mortality on day six following leaf dips in Knack solutions was higher than mortality of nymphs following leaf dips in water. Honeydew collected during the period between two to 50 h after leaf dip treatment had reduced amounts of glucose, fructose and trehalulose, but not sucrose and melezitose per nymph compared with honeydew from nymphs on leaves dipped in water. Results were more variable for sugars in honeydew collected 96 to 144 h after leaf dip treatment. Nebulizer sprays of Applaud and Knack to nymphs on cotton leaves also resulted in reduced amounts of sugars in honeydew and nymph mortality following treatments.

The coconut rhinoceros beetle, an economically important pest of coconut and oil palms, is effectively managed by application of its natural pathogen, the Oryctes nudivirus (OrNV), which act as a bioinsecticide. While this approach offers an environment-friendly alternative to chemical pesticides, the current method of production in infected larvae suffers from inconsistencies in virus productivity and purity. While the anchorage-dependent DSIR-HA-1179 insect cell line has been identified as a susceptible and permissive host for OrNV and therefore would be suitable for the in vitro mass production of the virus, no attempts have been made toward the mass production of the virus, because of the technological challenges that working with DSIR-HA-1179 cells represent. Thus, the main objective of this research was to develop processes for the in vitro production of OrNV in the DSIR-HA-1179 cell line. Knowledge of the growth kinetics and metabolic properties of the host cell line in a chosen culture medium, as well as the selection of an appropriate infection strategy, form the basis for the rational development of bioreactor-based virus production processes. However, characterization of these properties in the DSIR-HA-1179 cell line has been virtually precluded, due to its strongly adherent growth characteristics and the lack of a reliable method to accurately dissociate and count cells grown in monolayers. Using TrypLE™ Express enzyme, a technique allowing the precise counting of cells was developed. The cell line was adapted to grow in four serum-supplemented culture media: TC-100, IPL-41, Sf-900 II and Sf-900 III, which were then individually screened for cell growth and virus production in 25 cm2 attached T-flask cultures. TC-100 supplemented with 10% fetal bovine serum was chosen as a suitable culture medium, based on its capacity for achieving a high cell yield and OrNV production. The cell line metabolism was characterized with respect to nutrient consumption and metabolites production in this culture medium. Glucose, along with glutamine were found to be the nutrients that were consumed faster and to a greater extent, while other amino acids were not consumed to a significant degree. The production of metabolites was characterized by non-production of lactate and ammonia, and production of alanine, as a non-toxic alternative to ammonia. The influence of cell density (CD) at time of infection (TOI) and multiplicity of infection (MOI) on OrNV production was evaluated in T-flask cultures that were infected at different CDs at the TOI and a range of MOIs. The CD at TOI was found to significantly influence OrNV yields, while MOI influenced the dynamics of infection. The cell density effect was found to exist for the DSIR-HA-1179/OrNV system with the progressive decline in cell-specific yield beginning at low cell densities. It was found that in order to maximize OrNV volumetric yield, a combination of MOI and CD at TOI should be selected that allows to keep the maximum cell density reached by the infected culture within a range between 5.0 and 7.0 x 105 viable cells/ml. The roller bottle system was evaluated for its potential to scale-up DSIR-HA-1179 cell growth and OrNV production, and culture parameters were optimized for the improvement of cell and virus yields. An inoculum density of 3.3 x 105 cells/ml and culture volume of 60 ml resulted in the highest cell yield of 1.5 x 106 cells/ml, in 490 cm2 roller bottles. It was found that an optimal infection strategy for roller bottle cultures, which represented the most efficient use of viral inoculum, involved infecting cells at a density of 5.0 x 105 cells/ml and at a MOI of 1. The resulting OrNV volumetric yield of 2.5 x108 TCID50/ml, improved significantly the viral yields obtained in attached T-flask cultures infected under similar conditions (6.8 x 107 TCID50/ml). The microcarrier system was also evaluated for culturing DSIR-HA-1179 cells and producing OrNV in spinner flask bioreactors. Three types of microcarriers (Cytodex-1, Cytodex-3 and Cultispher-G microcarriers) were screened for their ability to support DSIR-HA-1179 growth. Cells attached to Cytodex-1 and 3, but failed to attach to Cultispher-G microcarriers. The final cell density reached in microcarrier culture was dependent on bead type and concentration, and the cell to bead ratio. At an optimal bead concentration of 1 mg/ml and cell to bead ratio of 30, cells grew to a maximum density of 1.7 x 106 cells/ml on Cytodex-1, but only to 1.3 x 106 cells/ml on Cytodex-3 microcarriers. Since it supported higher cell yields, Cytodex-1 was chosen to study the kinetics of OrNV production in this system. Microcarrier cultures infected at a cell density of 5.0 x 105 cells/ml and a MOI of 1, produced OrNV at 1.4 x 108 TCID50/ml, which was higher than the yield obtained in T-flask cultures infected under similar conditions. A framework of knowledge on the physiology, metabolism and growth kinetics of the DSIR-HA-1179 insect cell line has been developed in this thesis. In addition, the feasibility of using roller bottles and microcarrier systems for the in vitro production of the virus has been ascertained. It is envisaged that these findings will contribute to the future development of a large-scale industrial process for the production of the OrNV biopesticide.