Commercialisées depuis 1996, les plantes génétiquement modifiées produisant une toxine insecticide (toxine Cry) dérivée de Bacillus thuringiensis et appelées plantes Bt ciblent certains Lépidoptères ou Coléoptères ravageurs. Au fil des ans, les surfaces cultivées en plantes Bt sont de plus en plus importantes et contrôlent de larges populations d'insectes. Pour limiter le risque de développement de populations résistantes, une stratégie agricole appelée 'Haute Dose / Zone Refuge' est actuellement recommandée aux Etats-Unis par l'Environmental Protection Agency. Cette stratégie préventive nécessite la plantation d'une 'zone refuge' composée de plantes non-Bt utilisables par le ravageur ciblé et plantée à proximité de la 'zone Bt' qui synthétise une haute dose de toxine Cry. <p><p>Mon projet de recherche s’inscrit dans le cadre de l’évaluation de l'efficacité de cette stratégie et s’articule en deux phases :une phase expérimentale et une phase théorique. La première se concentre sur la caractérisation en laboratoire de l'impact des toxines Cry sur la biologie d'un ravageur. Cette phase constitue un support au volet théorique :la mise au point d’un modèle mathématique évaluant l'efficacité de la stratégie HD/R. L'originalité de ce projet repose entre autre sur l'interactivité entre ces deux volets.<p><p>Volet expérimental. Impact des toxines Cry sur la biologie de Plodia interpunctella. Nous évaluons séparément l'impact d'une gamme de concentrations de deux toxines Cry (CryXX et CryYY) sur une série de paramètres comportementaux et biologiques d'un insecte commun des denrées stockées: Plodia interpunctella (Hübner) (Lepidoptera :Pyralidae). Ces paramètres sont sélectionnés car leur variation pourrait avoir un impact sur l'efficacité de la stratégie HD/R dans le contrôle de la résistance. Il est donc pertinent de les quantifier pour intégrer dans le modèle des ordres de grandeur réalistes et générer des résultats qui ne sont pas uniquement basés sur des spéculations théoriques.<p><p>Volet théorique A. Efficacité de la stratégie HD/R pour des plantes Bt synthétisant une ou deux toxines simultanément. La stratégie 'HD/R' a été développée pour prévenir la résistance envers les plantes Bt synthétisant une seule toxine. Or, depuis 2003, de nouvelles variétés de coton Bt synthétisant simultanément deux toxines Cry sont commercialisées (BollgardII® et WidestrikeTM). Nous évaluons, grâce au modèle que nous avons développé, l'efficacité de cette stratégie lors d'une utilisation exclusive de plantes Bt synthétisant une ou deux toxines.<p><p>Volet théorique B. Impact du ralentissement du développement des insectes sur les plantes Bt sur l'efficacité de la stratégie HD/R. Le volet expérimental met en évidence un allongement de la durée du développement des larves se nourrissant sur une diète contaminée en toxine Cry. Ce ralentissement induit une séparation temporelle entre l'émergence des adultes de la zone Bt et de la zone refuge et perturbe une hypothèse principale de la stratégie HD/R: le croisement aléatoire entre adultes, indépendamment du génotype et de la zone d'origine. Dans ce troisième chapitre, nous étudions l'impact de la perturbation du croisement aléatoire sur l'efficacité de la stratégie HD/R. Nous testons également deux options pour optimiser la stratégie en cas d'asynchronie: l'utilisation de plantes Bt synthétisant une faible concentration en toxine (atténuant le décalage entre l'émergence des adultes) ou l'augmentation de la taille de la zone refuge (favorisant la survie des individus porteurs d'allèle de sensibilité et donc optimisant la dilution de la résistance à la génération suivante). <p><p>Ce travail s'intègre dans une problématique actuelle et utilise des outils de biologie théorique (théories de la dynamique et de la génétique des populations) ainsi que le développement d'un modèle mathématique. Il apporte des éléments de réponse et de réflexion sur l'optimisation de la gestion de la résistance des insectes mais c'est aussi une illustration de la complémentarité entre la biologie expérimentale et théorique.<p><p><p>/<p><p>On the market since 1996, genetically modified plants synthesizing an insecticidal toxin (Cry toxin) stemmed from Bacillus thuringiensis, called Bt plants, target several insect pests (Lepidoptera or Coleoptera). Bt crops cover increasingly larger areas and control important pest populations The Insect Resistance Management Strategy (IRM) strategy currently recommended in the U.S.A. to limit the development of resistant populations is the High Dose / Refuge zone (HD/R) strategy. This pre-emptive strategy requires a refuge zone composed by non-Bt plants, usable by the target insect and in close proximity of the Bt zone synthesizing a high toxin concentration.<p><p>My research project contributes to the effectiveness assessment of this HD/R strategy. It is structured on two main parts: an experimental, and a theoretical section. The first part characterizes the impact of Cry toxins on the biology of an insect pest. It is the basis of the theoretical part: the implementation of a mathematical model, which evaluates the effectiveness of the HD/R strategy.<p>The originality of this project is based on the interactivity of these two components.<p><p>Experimental section. Impact of the Cry toxins on the biology of Plodia interpunctella. We assess the impact of a range of concentrations of two Cry toxins (CryXX et CryYY) on several behavioural and biological parameters of a common pest of stored products: Plodia interpunctella (Hübner) (Lepidoptera :Pyralidae). These parameters are selected because their variation could influence the effectiveness of a HD/R strategy. So, it is important to quantify these parameters so that realistic values can be integrated in our model. The results of the model are thus not based on theoretical assumptions alone.<p> <p>Theoretical section A. Effectiveness of a HD/R strategy with Bt plants synthesizing one or two toxins. Initially, the HD/R strategy has been developed to limit the resistance towards Bt plants synthesizing one toxin. However, since 2003, new Bt cotton varieties synthesize two toxins simultaneously (BollgardII® et WidestrikeTM). We assess, with our model, the effectiveness of this strategy for Bt plants synthesizing one or two toxins.<p><p>Theoretical section B. Impact of the slowing down of the insect development reared on Bt plants on the effectiveness of the HD/R strategy. The experimental part demonstrates that larvae reared on a Bt diet have a protracted development duration. The consequence of this is a temporal separation between adult emergence in the two zones (Bt zone and refuge zone). This could affect the main assumption of the HD/R strategy, i. e. random mating independently of the genotype and of the native zone. In this third chapter, we study the impact of random mating disruption on the effectiveness of a HD/R strategy. We test two options to optimise the strategy in case of asynchrony: the use of Bt plants synthesizing a lower toxin concentration (limiting emergence asynchrony) or increasing the refuge zone size (favouring the survival of insect carrying one or two susceptible allele and thus optimising the dilution of resistance at the next generation). <p><p>This work is applied to a current issue. It uses some of the tools of theoretical biology (theories of population dynamics and population genetics) and develops a mathematical model. It provides some responses and some elements of thought about insect resistance management. It is also an illustration of the complementarity between experimental and theoretical biology.<p> / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

PhD (Zoology) / Department of Zoology / An ever growing human population and accelerating land use change is associated with the loss of species and their ecosystem services. Agricultural intensification has led to a worldwide threat of extinction to about one quarter of all bat species, despite the valuable ecosystem service of pest control provided by bats. The decline in bat populations is mainly attributed to the loss or fragmentation of habitats, roost sites and feeding opportunities related to agricultural intensification and land use change. Therefore, proactive management of bat communities in agricultural landscapes is essential. South Africa is the world’s largest producer of macadamias and the industry continues to grow. This study gains insight into the habitat use and foraging behaviour of insectivorous bat species on a temporal and spatial scale, in and around macadamia orchards in order to advise management strategies on how to increase bat activity and, possibly, pest control. It also focuses on the preferences of artificial roost sites used by insectivorous bats in macadamia orchards. The diet of insectivorous bat species is especially difficult to study and the least invasive tool to gain information is the study of bat faecal pellets. In order to provide evidence for the consumption of pest insect species by bats and thus incentive to farmers for a more integrated pest management approach (IPM), this study explored molecular approach to insectivorous bat diet analyses using fragment analysis of bat faecal pellets with fluorescent-labelled species-specific primers (designed for the CO I gene). This study was conducted in the subtropical fruit growing area of Levubu, Limpopo province, South Africa between the towns Thohoyandou (22°59'03.7 S, 30°27'12.8 E) and Makhado/Louis Trichardt (23°03'03.6 S, 29°55'12.7 E). Levubu also accounts for the second highest production of macadamia in South Africa. An introduction to the order Chiroptera and into the relevance of insectivorous bat species to agriculture as well as the importance of a more integrated pest management approach (IPM) focusing on bats is provided in Chapter One. Bats were acoustically monitored and light traps were used to catch arthropods during one annual cycle. I sampled five macadamia orchards once a month from September 2015 to August 2016 and used GIS and R to analyse both the general bat activity and foraging bat activity of the two main foraging guilds (open-air/clutter edge guild) in different land use types as well as total bat activity with respect to arthropod abundances. As reported in Chapter Two, results show that the overall clutter edge guild activity (number of passes) decreased with macadamia and orchard (all other fruit) cover in the macadamia high season (December to end of May) and increased with bush cover and distance to settlements (potential roosts) in the macadamia low season (June to end of November). Open-air guild activity increased with fallow cover (uncultivated grassland with scattered trees and shrubs) in the high season. Foraging activity (feeding buzzes) of the clutter edge guild increased with bush cover over the whole year. Total activity (both guilds) increased with abundance of true bugs (Hemiptera), including the main macadamia pests, and bush cover. Macadamia cover has a negative effect on the activity of the clutter edge guild in the high season, with low activity in the orchard center (high cover), and activity increasing in a linear way with decreasing orchard cover at the orchard edge (low cover). These results suggest that the clutter edge guild prefers foraging close to the edges of the orchards rather than in the center, while the open-air guild prefers semi-natural habitats (fallow). When numbers of pest arthropods drop in the macadamia orchards, the natural land use type, bush, becomes a more important foraging habitat and thereby increased the activity of the clutter edge guild. From June 2016 to July 2017, I scanned 31 bat houses, mounted on poles on six macadamia orchards, for bats or any other occupants such as wasps, birds and bees. Twenty-one multichambered bat houses of three slightly different chamber designs were erected on poles, in sets of three. Additionally, five bat houses of the type ‘Rocket box’, four bat houses in sets of two (black and white) and one colony bat house were erected. Bats were counted and visually identified to family or species level. From December 2016 to end of March 2017, three IButtons were installed to record temperature variation between one set of three bat houses. As reported in Chapter Three, results show that the central bat house in the set of three and the black bat house in the set of two had a significantly positive effect on bat house occupancy. There was a significant difference in the mean temperature between the houses in the set of three, with a significant difference in temperature of 0.46°C between the central and the first bat house. The three bat houses erected in sets varied slightly in their chamber design, with the central bathouse having the most chambers (six), while the bat houses to either side had less chambers (four), set at an angle or straight. This and the insulation to either side by the other bat houses is assumingly what caused the central bat house to be on average warmer. The Yellow-bellied house bat (Scotophilus dinganii) was by far the most recorded and the only species observed to co-habitat a bat house with another animal species, in particular honeybees. The study might confirm assumptions in that the microclimate of bat houses, respectively their insulation, sun exposure and color appear to be important factors influencing bat house occupancy. The two preferred bat houses in our study were the black, in the set of black and white, as well as the central, and on average warmest bat house, in the set of three. I collected bat faecal pellets with two different methods between July 2015 and April 2017 to determine the prevalence of pest insects in faecal pellets. Eighteen of the bat houses (in sets of three) on three different farms and two Egyptian slit-faced bat (Nycteris thebaica) roosts were fitted with trays in order to collect pellets from those occupied by bats. I noted occupancy of bat houses to species or family level to keep disturbance minimal. Additionally, I collected pellets from individuals captured by means of mist nets and harp traps. Four of the main pest-insects; the two-spotted stinkbug (Pentatomidae: Bathycoelia distincta), the green vegetable bug (Pentatomidae: Nezara viridula), the macadamia nut borer (Tortricidae: Thaumatotibia batrachopa) and the litchi moth (Tortricidae: Cryptophlebia peltastica), were collected from pheromone traps or after scouting for primer development and optimisation. After extracting DNA from the bat faecal samples the target regions were amplified in a multiplex PCR and fluorescently labelled PCR amplicons were analysed and interpreted. In order to verify multiplex analyses results, all samples were amplified with all four sets of primers in plates and those that produced amplicons were purified and sequenced. As reported in Chapter Four, results show that fragment analyses yielded a total of 63 out of 103 samples tested positive for pest insect species (61%) with a total of 92 positive fragments. Primer specificity could be confirmed to 100% for the sequences obtained for Bathycoelia distincta (26/26) and Nezara viridula (12/12) primers but not for all sequences obtained fot Cryptophlebia peltastica (18/30) and Thaumatotibia batrachopa (1/14) primers. One sample showed no positive fragments but contained a positive sequence for N. viridula. Three samples tested positive for one pest-species fragment but contained a positive sequence for a second pest-species (B. distincta, T. batrachopa and C. peltastica). Adding four positive fragments and one additional positive sequence to the data. This means that sequences of pest insect species were obtained from 54 out of the 103 samples (55.6%) with a total of 73 pest insect sequences. For the high season (December to end of May) a total of 37 positive fragments for the four pest insect species and 24 negative samples were yielded and for the a low season (June to end of November) a total of 36 positive fragments and 15 negative samples. Looking at the pest consumption of the different bat species or families, our results show that all of them foraged on pest insect species. Whereas, all species and families except Myotis bocagii and Rhinolophus simulator (for which N<2) foraged on both the Lepidopteran and Hemipteran pest species. Therefore, all families of bats of which faecal pellets were analysed for this study (Molossidae, Nycteridae, Rhinolophidae and Vespertilionidae) foraged on one or more of the four pest insect species. In summary, Chapter Five concludes that natural and semi-natural vegetation promote bat activity in macadamia orchards, and potentially bats' provision of the ecosystem service of pest control. In times of accelerating land use change, remnants of natural vegetation are important refuges and need to be maintained or restored to conserve bat species and promote their ecosystem services. The study also shows that bat activity might be improved by adding roosting opportunities to orchards. Warm and well-insulated bat houses mounted freestanding on poles and in sets appeared to work best in northern South Africa. Further research on co-habitation of bat houses and displacement behaviour as well as the potential importance of altitude and distance to water is needed. All of the species or families of bats from which faecal pellets were collected have been confirmed to forage on at least one of the four pest insects and the bat species have shown to be much more generalist and presumably opportunistic feeders than previously assumed. Thus, this study provides incentive and advice to farmers for a more integrated pest management approach (IPM). / NRF

Capsicum baccatum var. pendulum was first grown in the Makana District in 2005. Extremely little was known about best practices for cultivation or the insects and diseases associated with the crop in this area. The study was conducted during the second year of production, November 2005 and November 2006, in an attempt to identify the composition and phenology of insects occurring on C. baccatum. In the more rural parts of the Eastern Cape, and more particularly in Grahamstown, there are very few industries. With the advent of this new agricultural venture, a processing factory has been opened in Grahamstown creating more than 600 seasonal jobs in the factory and 1000 seasonal jobs on farms for local people. This business enterprise has not only brought about the creation of jobs, but also training and skills development and empowerment, generating much-needed income in this area. An extensive literature review yielded limited information on insect pests associated with Capsicum. Data from a pilot sampling trial undertaken were statistically analyzed to establish the number of plants to be scouted per site and the most effective scouting techniques to use. Based on the data available and insects collected during the pilot sampling trial, a surveillance programme was designed. Five different types of monitoring traps were placed in each of the eight study sites. Collection of trap catches and scouting of fifteen individual plants per site was undertaken on a weekly basis over the 52-week study period. The most commonly occurring potential insect pests were African Bollworm Helicoverpa armigera (Hübner), False Codling Moth Thaumatotibia leucotreta (= Cryptophlebia leucotreta) (Meyrick), Mediterranean Fruit Fly Ceratitis capitata (Wiedemann) and several species of thrips. Population densities of these pests and their phenology on Capsicum were determined. Statistical analyses established the efficacy of the monitoring traps for each pest, tested for differences among and between study sites, calculated an estimate of the number of pods damaged and a measure of plant damage.The results show that the majority of damage caused to the Capsicum baccatum cropping system was due to Mediterranean Fruit Fly populations. It was established that, although African Bollworm and False Codling Moth were present during the study period, their numbers were negligible and only nominal damage was caused by these pests. Damage caused by thrips species was apparent but not quantifiable. Intervention strategies using an Integrated Pest Management approach, are discussed.