Dissertations / Theses on the topic 'Eldana saccharina'

Thesis (PhD)--Stellenbosch University, 2013.<br>ENGLISH ABSTRACT: Two mathematical models are formulated in this dissertation for the population growth of an Eldana saccharina Walker infestation of sugarcane under the influence of partially sterile released insects. The first model describes the population growth of and interaction between normal and sterile E. saccharina moths in a temporally variable, but spatially homogeneous environment. The model consists of a deterministic system of difference equations subject to strictly positive initial data. The primary objective of this model is to determine suitable parameters in terms of which the above population growth and interaction may be quantified and according to which E. saccharina infestation levels and the associated sugarcane damage may be measured. The second model describes this growth and interaction under the influence of partially sterile insects which are released in a temporally variable and spatially heterogeneous environment. The model consists of a discretized reaction-diffusion system with variable diffusion coefficients, subject to strictly positive initial data and zero-flux Neumann boundary conditions on a bounded spatial domain. The primary objectives in this case are to establish a model which may be used within an area-wide integrated pest management programme for E. saccharina in order to investigate the efficiency of different sterile moth release strategies in various scenarios without having to conduct formal field experiments, and to present guidelines by which release ratios, frequencies and distributions may be estimated that are expected to lead to suppression of the pest. In addition to the mathematical models formulated, two practical applications of the models are described. The first application is the development of a user-friendly simulation tool for simulating E. saccharina infestation under the influence of sterile insect releases over differently shaped spatial domains. This tool provides the reader with a deeper understanding as to what is involved in applying mathematical models, such as the two described in this dissertation, to real-life scenarios. In the second application, an optimal diversification of sugarcane habitats is considered as an option for minimising average E. saccharina infestation levels, and as a further consequence, improving the cost-efficiency of sterile insect releases. Although many special cases of the above model classes have been used to model the sterile insect technique in the past, few of these models describe the technique for Lepidopteran species with more than one life stage and where F1-sterility is relevant. In addition, none of these models consider the technique when fully sterile females and partially sterile males are being released. The models formulated in this dissertation are also the first to describe the technique applied specifically to E. saccharina, and to consider the economic viability of applying the technique to this species. Furthermore, very few examples exist of such models which go beyond a theoretical description and analysis towards practical, real-life applications as illustrated in this dissertation.<br>AFRIKAANSE OPSOMMING: Twee wiskundige modelle word in hierdie proefskrif vir die populasiegroei van ’n Eldana saccha- rina Walker infestasie van suikerriet onder die invloed van gedeeltelik steriele, vrygelate insekte daargestel. Die eerste model beskryf die populasiegroei van en -interaksie tussen normale en steriele E. saccharina motte in ’n dinamiese, maar ruimtelik-homogene omgewing. Die model is ’n stelsel deterministiese verskilvergelykings onderhewig aan streng positiewe aanvangswaardes. Die primˆere doelstelling met hierdie model is om geskikte parameters te bepaal in terme waarvan die bogenoemde groei en interaksie gekwantifiseer kan word, en waarvolgens E. saccharina infestasievlakke en die gepaardgaande suikerrietskade gemeet kan word. Die tweede model beskryf hierdie groei en interaksie onder die invloed van gedeeltelik steriele insekte wat in ’n dinamiese en ruimtelik-heterogene omgewing vrygelaat word. Die model is ’n gediskretiseerde stelsel reaksie-diffusievergelykings met veranderlike diffusieko¨effisi¨ente onderhewig aan streng positiewe aanvangswaardes en zero-vloei Neumann-randwaardes op ’n begrensde ruimtelike gebied. Die primˆere doelstellings in hierdie geval is om ’n model tot stand te bring wat in ’n area-wye, ge¨ıntegreerde pesbestrydingsprogram vir E. saccharina gebruik kan word om die doeltreffendheid van verskillende steriele motvrylatingstrategie¨e te bepaal sonder om daadwerklik veldeksperimente uit te voer, en om riglyne daar te stel waarvolgens vrylatingsverhoudings, -frekwensies en -verspreidings bepaal kan word wat na verwagting na ’n onderdrukking van die pes sal lei. Bykomend tot die wiskundige modelle in hierdie proefskrif, word twee praktiese toepassings van die modelle ook beskryf. In die eerste toepassing word ’n gebruikersvriendelike simulasie hulpmiddel ontwikkel om E. saccharina infestasie onder die invloed van steriele insekvrylatings in verskillende ruimtelike gebiede te simuleer. Hierdie toepassing fasiliteer ’n dieper begrip van wat ter sprake is in die toepassing van wiskundige modelle, soos die twee modelle in hierdie tesis, tot werklike scenario’s. In die tweede toepassing word ’n optimale diversifisering van suikerriet habitats as ’n opsie vir die vermindering van die gemiddelde E. saccharina infestasie vlakke beskou, en gevolglik word die verbetering van die koste-doeltreffendheid van steriele insekvrylatings afgeskat. Alhoewel verskeie spesiale gevalle van die bogenoemde twee klasse van modelle reeds in die verlede gebruik is om die doeltreffendheid van die steriele-insektegniek te modelleer, beskryf weinig van hierdie modelle die tegniek vir Lepidopteriese spesies met meer as een lewensfase en waar F1-steriliteit ter sprake is. Verder beskryf geen van hierdie modelle die tegniek waar algeheel steriele wyfies en gedeeltelik steriele mannetjies vrygelaat word nie. Die modelle in hierdie tesis is ook die eerste waar die tegniek spesifiek op E.saccharina toegepas word, en waar die ekonomiese lewensvatbaarheid van die tegniek vir hierdie spesie oorweeg word. Verder bestaan daar min voorbeelde van soortgelyke modelle wat verder gaan as ’n teoretiese beskrywing en wiskundige ontleding na praktiese, werklike toepassings, soos in hierdie proefskrif ge¨ıllustreer.

The aim of the research presented in this dissertation was to further the implementation of push-pull for control of Eldana saccharina on sugarcane in the Midlands North region, KwaZulu-Natal, South Africa. Eldana saccharina, an indigenous stem borer, is the most damaging pest of sugarcane in South Africa, and sustainable control has still not been achieved. The push-pull strategy, a form of habitat management, has been developed for E. saccharina and is recommended as part of an integrated pest management (IPM) approach. Implementation of this strategy for both large- and small-scale farmers was facilitated through mixed methods social research. It included a novel exploratory network analysis to understand the process of technology adoption by farmers. Surveys showed that large-scale farmers have a good knowledge of E. saccharina, IPM and push-pull but that they needed more practical knowledge for implementation of the strategy. Farmers recommended experiential learning opportunities such as field days and model farms to get to know more about this technology. Despite demonstrating a positive attitude towards push-pull, farmers perceived it to be a ‘hassle’ and this is potentially the biggest barrier to its adoption. However, with suitable learning opportunities for farmers and good support for planting inputs, implementation of push-pull is likely to succeed. Sugarcane was shown to play an important role in the livelihoods and farming systems of small-scale growers. They did not perceive E. saccharina as a serious production constraint and had poor knowledge of the pest and its control. Extension for small-scale growers in this region should focus primarily on weed management and on reducing input costs, but still raising awareness of the increasing threat of E. saccharina. On-farm push-pull field trials showed a significant reduction of E. saccharina damage on two farms. Mean percentage damaged internodes decreased from 4.1% to 2.7% and from 1.7% to 1.1% in the presence of the repellent grass species, Melinis minutiflora. Where farmers did not manage their crops well, push-pull was not effective. It is therefore crucial that push-pull within an IPM framework be implemented together with good crop management practices. Stem borer surveys in wetlands on sugarcane farms revealed a high diversity of indigenous stem borers and parasitoids, including a stem borer species, Pirateolea piscator, which may pose a threat to crops in the future. These findings, together with a literature review on the significance of on-farm biodiversity and ecosystem services, demonstrated the value which wetlands have for pest management on sugarcane farms. Wetland health assessments were used to develop a tool for farmers to assess and utilise the wetlands on their farms for improved management of E. saccharina. This study highlights the importance of a farmer-participatory approach to implementation of knowledge-intensive farming practices such as push-pull. The importance of wetlands for providing pest regulatory services on sugarcane farms has shown that environmental sustainability needs to become a fundamental principle of farming and agricultural research. Participatory implementation of push-pull, as recommended in this dissertation, could act as a driving force for agroecology in the South African sugar industry and move sustainable farming practices off the pages of journals and manuals onto farmers’ fields.<br>Thesis (MSc (Zoology))--North-West University, Potchefstroom Campus, 2013

Thesis (PhD(Agric))--Stellenbosch University, 2012.<br>The sugarcane borer, Eldana saccharina Walker (Lepidoptera: Pyralidae) is the most limiting factor in the South African sugar industry with losses to this insect pest estimated to be at least ZAR60 million per annum. Because of its cryptic nature as well as the fact that E. saccharina is both indigenous to Africa and occurs on several host plants, attempts to control or eradicate it using several available methods have not been very successful. However, the sterile insect technique (SIT) is one of the newer control methods that can be incorporated into an area-wide integrated pest management (AW-IPM) programme to achieve better control or eradication. The implementation of the SIT program needs to go through a series of well-researched phases in order to be successful. In the first of this multi-phase project, it was determined that E. saccharina is susceptible to ionizing radiation, and is thus a suitable candidate for the SIT development against it and that a sub-sterilizing dose of 200 Gy is sufficient to induce F1(inherited) sterility in male and complete sterility in female moths respectively. The results presented here are discussed in the context of further development of the SIT as an addition to the arsenal of tactics in an AW-IPM programme against E. saccharina. Based on these initial findings, the study examined the lek and mating behavior of male moths subjected to three radiation doses (150, 200, 250 Gy) against normal non-irradiated/fertile moths. Both mass-rearing and irradiation of E. saccharina led to a quantitative departure of male mating behavior away from that exhibited by their wild counterparts. However, treated males are still able to form leks and mate with wild females. Male E. saccharina irradiated at all three doses tested were found to be as active and competitive as wild males, but in some of the traits measured, performance diminished significantly with an increase in the radiation dosage. In general, the performance of moths treated at 200 Gy did not differ significantly from that of moths treated at 150 Gy and therefore the former dose is ideal for SIT development since it results in a lower residual F1 fertility than the latter. The level of mating competitiveness and compatibility was assessed under both laboratory and semi-field conditions in pairwise comparisons consisting of laboratory reared vs. wild (L-W), 200 Gy irradiated vs. wild (S-W) and laboratory reared vs. irradiated moths (L-S). Based on the results from the more robust field cage assays, the mating indices generated indicated that the mass-reared E. saccharina strain produced in South Africa has not yet evolved sexual behaviours suggestive of incipient pre-mating isolation barriers with local wild strains. Wild moths did not discriminate against either the partially sterile or laboratory reared moths and most importantly, the irradiated males mated significantly more than their wild counterparts regardless of the type of female. The irradiated insects could therefore achieve the purpose for which they are intended upon release into the field. Third, the critical thermal limits (CTLs) to activity at high and low temperatures (i.e. critical thermal maxima “CTmax” and minima “CTmin”) of different E. saccharina strains/treatments were investigated under standard experimental conditions. The effect of laboratory rearing and increasing radiation dosage on thermal tolerance of the adult stage of E. saccharina was explored. There were highly significant differences between the laboratory-reared and wild strain and also between non-irradiated and irradiated strains in both CTmax and CTmin. Laboratory reared E. saccharina moths were more heat tolerant compared to wild moths for both genders while in the case of CTmin, the reverse was true. Irradiation had a negative effect on both CTmax and CTmin. Moths treated at the lowest radiation dose were more cold and heat tolerant than those treated at higher dosages thereby reinforcing the importance of lower dosages rather than those that induce full sterility against E. saccharina. In general, gender effects on the CTLs were non-significant. Pilot sterile male releases in shade house trials to measure the impact of sustained releases of partially sterile adult males at an over-flooding moth ratio of 10T: 1U (treated to untreated),were conducted to measure their efficacy to stop E. saccharina incursions and suppress populations prior to testing in pilot studies under true season-long and area wide conditions. Results from the current study demonstrated that releasing partially irradiated (200 Gy) adult male moths at the afore mentioned release rate significantly reduced sugarcane stalk damage as well as lowered the number of fertile progeny from F1to succeeding generations in a stable E. saccharina population initiated in a cage house. There were more damaged internodes per stalk in the control than in the sugarcane receiving regular releases of partially sterile male moths. Overall, there were significantly more undamaged stalks in the treated sugarcane than the untreated control. Furthermore, there were significantly more larvae per stalk retrieved from the control compared to the treated sugarcane suggesting that the sustained release of steriles was efficacious in reducing emergence of fertile larvae in the succeeding generations. The results of this study indicate that there is considerable scope for the SIT against E. saccharina.

Eldana saccharina, an indigenous lepidopteran stemborer, is considered the most damaging pest of sugarcane in South Africa. Researchers have advocated the use of an area-wide integrated pest management (AW-IPM) programme as a means of improving the control of this pest. A push-pull strategy was developed as a component of this AW-IPM approach. The push-pull strategy in sugarcane is a habitat management method of pest control that uses plants that are both repellent (Melinis Minutiflora) and attractive (Cyperus dives, Cyperus papyrus and Bt-maize) to E. saccharina. Previous research into push-pull has shown that this strategy is an effective tool for the control of E. saccharina. Push-pull has been implemented successfully in the Midlands North sugarcane growing region of KwaZulu-Natal (KZN), South Africa. Despite the proven efficacy of push-pull, very little push-pull research has been conducted in the coastal sugarcane growing regions of KZN, and adoption of this technology has been poor in these regions. Therefore, the aim of this research was to facilitate the implementation of push-pull for the management of E. saccharina in sugarcane in the coastal regions of KZN. This was done by focussing on on-farm field trials and farmer participatory research. On-farm push-pull field trials were conducted on five model farms in the North and South Coast sugarcane growing regions of KZN. High levels of E. saccharina were recorded during this study. The push-pull treatment sites showed a significant reduction of E. saccharina damage on four of the five farms used in the study. Mean percentage of stalks damaged decreased by up to 50 % in the presence of the repellent grass species, M. minutiflora. The number of E. saccharina found per 100 stalks also decreased significantly at these farms. The farm which did not show a significant reduction in E. saccharina populations or damage had low numbers of this pest in the sugarcane throughout the experiment. This demonstrates that push-pull is more effective in areas that have high levels of E. saccharina. Stem borer surveys in wetlands on sugarcane farms revealed that high numbers of E. saccharina were found within the pull plants, C. papyrus and C. dives, in comparison to the push-pull sites. This verifies that the pull plants do work efficiently to attract E. saccharina away from sugarcane. Additionally, eight parasitoids emerged from E. saccharina larvae collected in wetland sedges. The beneficial roles that push-pull plants play in attracting and maintaining natural enemies in the agroecosystem are discussed, and these findings further demonstrate the important ecosystem, and pest management services that wetlands provide on sugarcane farms. The success of the push-pull trials in this study show that this technology can be an effective tool for controlling E. saccharina in the coastal sugarcane growing regions. The timing of the planting of push-pull plants was shown to play a role in the efficacy of this technology. The study also confirmed that push-pull should be used as a component of AW-IPM in conjunction with good crop management practices. Surveys were undertaken to determine large-scale sugarcane growers' (LSGs) knowledge and perceptions of E. saccharina and other pests. Research regarding the farmers' perceptions of push- pull was also conducted to better understand the drivers and barriers to adoption of push-pull, and other new technologies. The surveys found that large-scale farmers in the coastal regions suffer from high infestations of E. saccharina. As such there is scope for the introduction of new pest management practices such as push-pull in this area. Farmers also demonstrated a good basic knowledge of E. saccharina and IPM. However, LSGs had a poor understanding of push-pull and how it works, as well as the plants that make up the push-pull system that is being implemented against E. saccharina in South Africa. A dearth in practical knowledge regarding the implementation of push-pull was seen as a major barrier to the adoption of this strategy, as was financial instability, farmer attitudes and poor institutional support. Farmers recommended collaboration amongst stakeholders, improved education, proof of the efficacy of push-pull and incentives as tools to improve the implementation of this strategy in the coastal sugarcane growing regions of KZN. Farmers preferred direct contact with extension personnel and experiential learning opportunities when acquiring information about push- pull and other new pest management practices. If opportunities for push-pull education are increased through direct contact with extension personnel, and through on-farm demonstrations, and if inputs are provided in the form of push-pull plants, it is likely that push-pull will succeed amongst coastal LSGs, especially since farmers had an overall positive attitude towards the technology. Surveys amongst small-scale sugarcane growers (SSGs) showed that sugarcane is important in the lives of these farmers. The SSGs perceive pests to be a major constraint to their farming systems, and they identified E. saccharina as a major pest of sugarcane. The farmers also demonstrated good knowledge of sugarcane pests and vegetable pests. However, SSGs lacked knowledge regarding pest management practices and beneficial insects. Extension and advisory services should to continue concentrating on pest management practices to educate SSGs on the variety and application of pest control strategies. SSGs were found to employ complex, diverse and integrated agricultural systems that are well-suited to the implementation of IPM technologies such as push-pull. Since insect pests act were found to be a major constraint to SSG sugarcane production, push-pull was deemed a feasible pest management strategy for coastal farmers and its implementation by SSGs should be further explored. SSGs in this study were also concerned about vegetable pests, therefore if push-pull can be adapted to help protect additional crops, adoption of this technology by small-scale growers will improve.

Host recognition behaviour of two braconid larval parasitoids Cotesia sesamiae and Cotesia flavipes was studied using suitable stemborer hosts [i.e. Busseola fusca for C. sesamiae, and Chilo partellus for C. flavipes] and one non-host [Eldana saccharina]. The wasps displayed similar sequences of behavioural steps when locating their hosts largely depending on their antennae for host recognition and both antennae and tarsi for final host acceptance and oviposition. Tactile and contact chemoreception stimuli from the hosts seem to play a major role in oviposition decision by the parasitoids. In addition, the external morphology and distribution pattern of sensilla present on antennae, tarsi and ovipositor of the parasitoids were examined by scanning electron and optic microscopy after staining with silver nitrate. Three sensillar types were identified on the distal antennomeres: (i) non-porous sensilla trichoidea most probably involved in mechanoreception, (ii) uniporous sensilla chaetica likely to be gustatory and, (iii) multiporous sensilla placodea likely to be olfactory. The tarsi possess a few sensilla chaetica which could be gustatory while the manubrium is likely to be used in detection of vibrations. The distal end of the ovipositor bears numerous multiporous dome-shaped sensilla. Additionally, the ability of the wasps to discriminate between contact cues was studied. When host larvae were washed in distilled water the wasps did not insert their ovipositors. However, ovipositor insertion resumed when washed host or non-host larvae were painted with water extracts of their respective host larvae. The water extracts of the suitable hosts were more attractive to the wasps than those of non-hosts. Similarly, the frass is important in host recognition during short-range examination as those of respective hosts are more intensely antennated than of non-hosts. The parasitoids were able to discriminate the regurgitant of E. saccharina by not antennating the cotton wool ball of this host; while the regurgitant of B. fusca and C. partellus appeared not useful in discriminating between the two species for both parasitoid species. Further analysis suggests the presence of a protein(s) component(s) in the regurgitant possibly responsible for host recognition and oviposition by C. flavipes.<br>Thesis (Ph.D. (Environmental Science))--North-West University, Potchefstroom Campus, 2009.

Le maïs cultivé en zones de savanes de Côte d'Ivoire est attaqué par 5 espèces de Lépidoptères foreurs qui s'attaquent soit a la tige, soit a l'épi, soit aux deux. Étude de la chétotaxie larvaire des différents foreurs, de l'habitus des stades préimaginaux, de la morphologie et de la biologie de Mussidia nigrivenella. Résultats de l'étude des fluctuations des populations de ces foreurs. Mise en évidence de l'efficacité de la deltaméthrine en concentré émulsifiable a 15 grammes de matière active à l'hectare. Iimpact des divers insectes sur le rendement et calcul d'équations de régression reliant la perte de rendement a l'attaque

Eldana saccharina Walker (Lepidoptera: Pyralidae) is an indigenous insect widely distributed throughout Sub-Saharan Africa that is a major pest of sugarcane in southern Africa. Studies have shown that populations from West Africa have distinct behavioural differences compared to populations from eastern and southern Africa. In addition, the parasitoid guilds attacking populations in these regions are markedly different. This marked behavioural and parasitoid guild variation evoked a hypothesis of genetic diversification. To evaluate this hypothesis a project on the phylogeography of E. saccharina was initiated. The project was planned to include sampling of as many regions as possible in its known range in Africa, to obtain specimens of E. saccharina for genetic analysis. When these surveys were initiated in Ethiopia, it was found that there was no published literature available on the occurrence of stem borers in Ethiopian sugarcane. It was thus clear that no stalk borer/parasitoid surveys had been completed in either sugarcane or any large grass and sedge indigenous hosts in Ethiopia. The study was thus expanded beyond the investigation of only the genetic diversity of E. saccharina, to include area-wide surveys to determine ecological aspects of the borer complex in suspected host plants, including sugarcane, in Ethiopia. In this way the host plant range and distribution of E. saccharina and other sugarcane borers in Ethiopia in particular could be determined, samples for a larger phylogeography project could be collected, and the insect's impact on sugarcane could be assessed. Quantified area-wide surveys of the sugarcane estates and small-scale farmer fields of Ethiopia were conducted between December 2003 and February 2004. The surveys verified the presence of four lepidopteran stem borer species on Ethiopian sugarcane. These were Chilo partellus Swinhoe (Lepidoptera: Crambidae), Sesamia calamistis Hampson (Lepidoptera: Noctuidae), Busseola fusca Fuller (Lepidoptera: Noctuidae) and Busseola phaia Bowden (Lepidoptera: Noctuidae). The surveys indicated that Busseola species are the major and most widely distributed sugarcane stem borers in sugarcane farms of Ethiopia. Over 70% of the peasant sugarcane fields visited were infested by these borers, with the highest levels of infestation (35% and 50%) being in the northern and western part of the country, respectively. Busseola was also the predominant stem borer of sugarcane in two of the three estates (Wonji and Finchawa). Chilo partellus and S.calamistis were recovered in very low numbers at all the commercial estates and from peasant farms in the western part of Ethiopia. However, C. partellus was the predominant sugarcane stem borer in lowland areas of northern, southern and eastern parts of the country. Eldana saccharina was recovered from large sedges in waterways of Metehara and Wonji sugar estates in the central part of the country, and sedges growing around lakes in northern and southern Ethiopia, but not from sugarcane anywhere in Ethiopia. The phylogeographic study conducted on E. saccharina populations from eleven countries of Africa clearly showed the population structure of the insect within the continent. Five hundred and two base pairs of the mitochondrial DNA (mtDNA), corresponding to the Cytochrome Oxidase subunit I (COl) region, were sequenced to clarify phylogenetic relationships between geographically distant populations from eastern, northern, southern and western Africa. Results revealed that E. saccharina is separated into four major populations corresponding to their geographical location, i.e. West African, Rift Valley and two southern African populations. Sequence divergence between the four populations ranged from 1% to 4.98%. The molecular data are congruent with an isolation by distance pattern except for some of the specimens from eastern and southern Africa where geographically close populations are genetically distant from each other. Geographical features such as the Rift Valley and large water bodies in the continent seem to have a considerable impact on the genetic diversity in E. saccharina. Identification of field-collected stem borer specimens was done using classical taxonomic techniques, except for Busseola spp. where DNA barcoding was used. As field-collected larval material of Busseola died before reaching the adult stage, identification of species using adult morphology was not possible. 'Sequence divergence in the COl gene was used as a tool to identify the species of Busseola attacking Ethiopian sugarcane. Partial COl sequences from Ethiopian specimens were compared with sequences of already identified noctuid species from the East African region. Results of the sequence analysis indicated that the Busseola species complex in Ethiopian sugarcane comprised B. fusca and B. phaia. Sequence divergences between Ethiopian Busseola species was as high as 5.0 %, whereas divergences within species were less than 1% in both species identified. Several larval parasitoids, bacterial and fungal diseases of stem boring caterpillars were also recorded in Ethiopian sugarcane. Amongst these was Cotesia flavipes Cameron (Hymenoptera: Braconidae). This exotic parasitoid has been introduced into several African countries for the control of C. partellus in maize and sorghum, but had never been released in Ethiopia. To investigate the origin of C. flavipes in Ethiopian sugarcane, molecular analyses were conducted on Ethiopian specimens from sugarcane and specimens of C. flavipes from different countries of Africa released from the Kenyan laboratory colony, again using COl sequences. Results of the analysis revealed that the C. flavipes population that had established in sugarcane fields of Ethiopia was similar to the south east Asian populations released against C. partellus in maize in other parts of Africa, and different from other populations of this species, providing evidence that the Ethiopian C. flavipes is likely to be a descendant of the original Pakistani population that was released in different parts of Africa. The study reveals the importance of lepidopteran stem borers in sugarcane production in Ethiopia and highlights the role of molecular methods in species identification and determining phylogenetic relationships. More importantly, this study establishes the continental phylogeographical pattern of the indigenous moth, E. saccharina. The impact of geological events, geographic barriers and cropping systems on the evolution, distribution and abundance of stem borers are discussed. Future areas of research for understanding more about the phylogeographic relationships of E. saccharina and management of stem borers are discussed.<br>Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

Since becoming a pest in graminaceous crops in Africa, the African sugarcane stalkborer Eldana saccharina Walker has been the subject of much study. Its very cryptic habits have precluded more commonly available control measures being used against it. Biological control is regarded as a viable control option, but the apparent paucity of parasitoids in graminaceous crops leads to E. saccharina being regarded as lacking parasitoids, and thus not a good candidate for biological control in the classical sense. In contrast, this project argues that interactions in indigenous hosts of E. saccharina had been ignored, and that classical biocontrol principles and basic ecological theory could be applied by the discovery, collection and introduction of parasitoids from its indigenous habitat to its newly adapted habitat, sugarcane. The habitat offered by Cyperus papyrus L. was shown to be heterogenous both temporarily and spatially. Umbels, from young through mature to senescent, were available in the same proportion for colonisation throughout the year. Umbels with sexual reproductive stages (seeds) were present from early spring into late summer, and provided an additional component to the already heterogenous environment. Young umbels, in addition, developed from rhizomes in an environment regarded as sub-optimal for photosynthesis, until they reached the canopy. All stages of umbels were attacked by E. saccharina, but larvae were only found in rays of umbels and in the apex of the culm, which was the meristematic area for rays, both high nutrient areas. Young umbels were never found with borer pupae, only smaller larvae, indicating that E. saccharina development matched growth of young umbels until they reached canopy height. Also, the majority of borings found were occupied, indicating that infestation of young umbels was recent. All stages of E. saccharina development were found in mature umbels, which were also most abundant at anyone time. Numerous empty borings were found in addition to those occupied, indicative of past occupation by E. saccharina. Very few young larvae were found in old umbels, the majority of life stages found being pupae or empty pupal cases, and also many empty borings, showing that old umbels were not suitable for E. saccharina development. A guild of parasitoids which comprised Orgilus bifasciatus Turner, the most common parasitoid of small and smaller medium E. saccharina larvae, Goniozus indicus Ashmead the most common parasitoid of larger medium and large larvae, and an entomogenous fungus Seauveria bassiana (Sals.) Vuill. attacking all life stages of E. saccharina was found. Three uncommon parasitoids of smaller saccharina were also found, viz. Sassus sublevis (Granger), Iphiaulax sp. and Venturia sp. The former three natural enemies were instrumental in depressing a major outbreak within two months of it being observed and then maintaining the host population at a lower level in C. papyrus. G. indicus and B. bassiana were most effective during the summer and autumn months, and O. bifasciatus most effective during the winter months. This study supports the hypotheses that the apparent paucity of parasitoids and lack of biological control success thus far against E. saccharina in sugarcane has been because very little was known about its ecology and biology in its numerous indigenous host plants, and that studies of the latter factors coupled with ecological theory could enhance biological control programmes against this borer. As more indigenous host plants are investigated in the same way as has been done with C. papyrus, more will become known of natural enemies of E. saccharina. Parasitoid guilds could be selected, even from rare parasitoids in the more stable indigenous habitats, which would provide control in the unstable habitat of sugarcane.<br>Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 1994.

The eldana stalk borer (Eldana saccharina Walker) is the most serious pest of the Southern African sugarcane industry, and it is imperative that effective control measures are available to minimize economic damage. Because conventional control methods have had limited success, cultivar resistance is seen as the most viable method of controlling infestation. However, due to the space- and time-consuming nature of the present screening methods, only small numbers of cultivars can be tested relatively late in the Plant Breeding selection programme. Increased resistance in breeding and selection populations is therefore slow. Buds are a preferred entry point of eldana larvae as they are softer than the rind that is present on the rest of the stalk surface. Preliminary results by other workers suggested that near infrared spectroscopy (NIRS) could provide a rapid screening method for the chemical profile in bud scales, the outer coating of buds and therefore the first contact point of an invading larva. If feasible, analysis of samples using this method could be done in the South African Sugar Experiment Station's (SASEX) stage two selection trials, providing an early indication of eldana resistance on large numbers of cultivars, without the necessity of separate trials. However, knowledge of how environments, position of bud scales on the stalk and age affect NIRS is required in order to determine the feasibility of the method. Planting of a trial with an identical set of genotypes across a range of environments, sampled at a number of ages, would provide the necessary information on environmental effects, whilst simultaneously providing the necessary range of samples to develop a calibration between bud scale chemical profiles and eldana resistance ratings. Inheritance patterns of the characteristics being measured is also required if they are to be used in a breeding programme. The original work by Rutherford (1993) was carried out on only five calibration sets (a set of standard clones with relatively well-known eldana resistance ratings), and different sets were not comparable due to what was assumed to be environmental differences between calibration sets. One aspect of the current experiment was to examine more closely the effect of genotype x environment interaction (G x E) on the performance of the NIRS technique under a range of conditions. Two sites were chosen to represent the conditions encountered in trials carried out by SASEX. The crops were sampled at three ages, representing the range of ages at which sugarcane is harvested in South Africa. Two locations on the stalk were also examined, top and bottom, for removal of bud scales, based on the assumption that aging of bud scales may affect chemical composition. A new NIRSystems 6500 instrument was acquired during the course of this study. Data from the new instrument indicated that there were no longer differences between the different calibration sets, and therefore no longer differences between environments. Spectra for different samples were very close, the differences being of the same scale as those recorded with repeated measures of the same samples, or between the readings for the standard solvent solution. This led to the conclusion that the differences observed on the original NIRSystems 5000 instrument were due to instrument error, not environmental differences. More importantly, the different calibration sets were not comparable despite being similar to each other. Prediction from one calibration set to another was low. These observations led to the conclusion that NIRS was not a suitable method for determining chemical compounds associated with tolerance of sugarcane genotypes to eldana borer. The original NIRS instrument was subject to error, and the small number of calibration sets included in the study led to the erroneous conclusion that NIRS was suitable for the prediction of varietal tolerance to eldana. With the acquisition of the new instrument, the errors generated by the old instrument became apparent. With the increase in number of calibration sets included in the study, it also became apparent that a global calibration covering all environments was not possible. An analysis of the heritability of the chemical compounds associated with eldana resistance was also included in this study. A biparental progeny design of 24 crosses with 33 unselected offspring per cross was used. This trial would have been analysed once the calibration had been developed using the environmental trial, and it would have provided knowledge of the breeding behaviour of the chemical compounds associated with tolerance to eldana. Because the NIRS technique proved to be unsuitable for detection of chemical compounds associated with eldana resistance, the heritability of these chemical compounds could not be studied. As the NIRS study did not produce data, the G x E interaction analysis and determination of heritability was applied to the bud scale mass data set. This study showed a relatively low positive correlation between bud scale mass and resistance to eldana. The broad sense heritability estimate for bud scale mass from the G x E interaction analysis was 0.45, and the narrow sense heritability estimate from parent-offspring regression analysis was approximately 0.27, suggesting a low degree of genetic determination in bud scale mass. The G x E interaction analyses gave varying results depending on the method used. The ANOVA analysis suggested that ages, sites and years had an effect on bud scale mass, while deviation from maximum plot showed no significance for G x E interactions. The number and choice of genotypes selected as unstable also varied with the method used to determine the stability of individual genotypes. Regression analysis and rank order analysis revealed a number of unstable genotypes, whilst stability variance and ecovalence, which produced similar results, detected only two unstable genotypes. In the rank order analysis correction of data to remove genotype effect, reduced the number of unstable genotypes, suggesting that the G x E interaction effect was partially confounded with the bud scale mass of the genotypes. This was a more reliable method than the uncorrected rank order analysis, and would be the preferred analysis type of all those tried.<br>Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.

Commercial sugarcane in South Africa is a monoculture, and therefore lacks vegetation diversity, which is instrumental in increasing associated faunal diversity. Diverse habitats tend to support more stable populations of herbivorous and predacious animals. It is hypothesised that lack of this diversity in sugarcane was partially responsible for the existence of Eldana saccharina infestation levels that are higher than is commercially acceptable. Amongst the available E. saccharina control strategies, is habitat management. This has been developed with the view of increasing and enhancing predator foraging activity. Through increasing arthropod predator abundance and activity, it is believed that E. saccharina control may be enhanced. In this study, habitat diversity was increased through intercropping beans within sugarcane. Arthropod populations were monitored throughout the sugarcane-growing period, to determine what effect this intercropping had on known potential arthropod predator populations of E. saccharina. The study site was divided into two plots: the intercrop (beans planted within sugarcane rows: sugarcane-bean intercrop) and sole sugarcane: control plot. At monthly intervals, epigeal arthropods were sampled with pitfall traps, while foliage associated arthropods were sampled with a suction trap. Predator activity at the base of the sugarcane stalk, where E. saccharina lays its eggs was monitored with sticky traps. Sampling took place in the sugarcane-bean intercrop and control plots as well as in the roadway bordering the study site. Epigeal predator habitat preference was assessed by randomly placing pitfall traps in the sugarcane rows, bean rows, interrows between sugarcane rows, interrows between sugarcane and bean rows and the roadway. Corresponding with monthly trapping, an E. saccharina infestation and damage survey was conducted. Environmental factors such as weather, light intensity, plant (beans and sugarcane) phenology and weed density were measured, and their effect on E. saccharina potential arthropod predators examined. At harvest, sugarcane stalks were sampled for sucrose yield analysis. Potential E. saccharina predators that were captured included species of the orders and/or families Acarina, Blattidae, Formicidae (Pheidole megacephala and Dorylus helvolus) and Araneida (Lycosidae, Oxyopidae, Thomisidae and Salticidae). P. megacephala and species of Acarina were the only predators caught with all three trapping techniques, thus indicating that they occurred both on the ground and foliage. D. helvolus and Acarina were the only predators caught in significantly higher numbers in the intercrop, suggesting that increased habitat management had positively affected their population sizes. D. helvolus were captured both on the ground and length of sugarcane stalk, while species of Acarina were captured on the ground, foliage and at the base of sugarcane stalk, indicating that they forage at the base of the stalk, where E. saccharina activity is concentrated. Specific ground habitats preferred by D. helvolus included the sugarcane rows and bean rows, while Acarina preferred the interrows between sugarcane and bean rows. Despite the generally low E. saccharina infestation levels during this study, significantly higher levels of infestation occurred in the intercrop when compared to sole sugarcane. As expected with high infestation, higher (although not statistically significant) damage occurred in the intercrop. Surprisingly, sucrose yield and sugarcane stalk mass were slightly higher in the intercrop. The implications of the observations made during this study are discussed in the context E. saccharina management.<br>Thesis (M.Sc.)-University of Durban-Westville, 1998.

The South African Sugarcane production is one of the world’s leading sugarcane (Saccharum spp. Hybrid) producers. However, in recent years Eldana saccharina Walker has been the most destructive pest in South African sugarcane production, causing great crop loses per annum and is the most important factor limiting sugarcane productivity. The pest has been monitored using a traditional visual approach whereby a representative sample of stalks is taken from a field and split longitudinally to assess damage and count the number of E. saccharina larvae and pupae. However, this approach is time-consuming, labour intensive and sometimes biased as only easily accessible areas are often surveyed. In order to investigate a more economical but equally effective survey methodology, this study aimed to determine the potential of using hyperspectral remote sensing (spectroradiometry) for identifying sugarcane attacked by E. saccharina. A hand-held spectroradiometer ASD Field Spec® 3 was used to collect leaf spectral measurements of sugarcane plants from a potted-plant trial taking place under shade house conditions at the South African Sugarcane Research Institute (SASRI). In this trial, nitrogen (N) and silicon (Si) fertilizers were applied at known levels to sugarcane varieties. Varieties were either resistant or intermediate resistant or susceptible to E. saccharina attack. In addition, watering regimes and artificial infestation of E. saccharina were carefully controlled. Results illustrated that severe E. saccharina infestation increased spectral reflectance throughout the whole spectrum range (400 – 2500 nm) and caused a red-edge shift to the shorter wavelength. Eldana saccharina stalk damage was also linearly related to modified normalized difference vegetation index (mNDVI) using R2025 and R2200 (R2 = 0.69). It was concluded that hyperspectral data has a potential for use in monitoring E. saccharina in sugarcane rapidly and non-destructively under controlled conditions. A followup study is recommended in field conditions and using airborne and/or spaceborne hyperspectral sensors.<br>Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2009.

Petri dish and olfactometer tests demonstrated that Goniozus natalensis (Gordh) females exhibit a host-searching response upon contact and at a short distance by olfaction, to a kairomone in the frass of its natural host Eldana saccharina (Walker). The host-searching response was found to be elicited by E. saccharina frass from a range of substrates, namely: two host plants of E. saccharina, papyrus and sugarcane, and four media: sugarcane, papyrus, and cellulose based media and a synthetic medium containing no plant material. The host-searching response was not elicited by Sesamia calamistis (Hamps) sugarcane medium frass. The sexual state and age of G. natalensis females were found to influence the host-searching behaviour. Mated females showed the behaviour in the petri dish bioassays only after completing their preoviposition time of two to three days, whilst virgin females took longer, even though their preoviposition time was found to be the same. The response to male or female produced E. saccharina sugarcane frass was not statistically different, nor was there a statistically significant preference for either frass type, given the choice. Four way olfactometer tests showed that an E. saccharina sugarcane frass odour elicited a host-searching behaviour in mated two to three day old G. natalensis females. Various solvents were tested for their ability to isolate the kairomone from E. saccharina sugarcane frass. Chloroform proved to be the best solvent when tested in petri dish and olfactometer bioassays. The preliminary results of the GC/MS analysis of the chloroform extract of E. saccharina sugarcane frass are presented. The chemicals identified are compared with chemicals identified as host location kairomones for other insect parasitoid-host studies.<br>Thesis (M.Sc.)-University of Natal, 1990.

The complexity of the behaviour of neonate Eldana saccharina Walker (Lepidoptera: Pyralidae) larvae and the limited information on their response to the morphological characteristics of South African sugarcane varieties was the primary justification to study antixenotic/antibiotic effects on larval behaviour. Laboratory experiments were conducted with stalk segments in plastic jars inoculated with larvae and in a metal cage covered with gauze. In jars, the larvae were observed until they penetrated the stalks. After 14 days, the stalks were dissected and larvae weighed. In all varieties, larvae moved directly to the node after inoculation and penetrated the stalk through leaf scars and buds. No significant differences in larval mass were observed among varieties. In cage experiments different parts of the node, namely the rind below the wax band; the bud; and the root primordia were tested. There was a clear indication that rind hardness and the budscale properties are associated with varietal resistance and only affect early instars. The experiments were repeated using whole cane plants in a glasshouse. The results were similar to those of laboratory experiments. In the Insect Rearing Unit, scraped waxes from different varieties were incorporated into the diet. Larval masses from different diets showed significant differences among varieties, but they did not conform to the known resistance ratings, as cane varieties N12 and N21 showed high susceptibility, instead of resistance. Dispersal behaviour of neonates shortly after hatching was investigated in 'mobility experiments' conducted on live cane plants. Mobility is important because the more time neonates spend wandering around on the stalk surface or on exposed parts of the plant, the more vulnerable they are to predation and other adverse factors that may reduce their survival. Experiments to test stalk penetration by larvae on the node showed that neonates required a softer food source before attacking the hard nodal parts. Second and third instar larvae were used subsequently to the mortality of all neonates fed on the rind, which in turn resulted in non-significant differences, suggesting that feeding on debris and/or leaves is critical to the survival and penetration of larvae into the sugarcane stalk. Incorporation of the characteristics tested in these experiments aims to reduce the number of larvae that penetrate the stalk and to expose them for longer on the surface where their numbers may be controlled by predators and insecticides. The resistant varieties used in these experiments have high fibre and less sugar, but newer varieties, such as N29 and N33 incorporate both high resistance and high sucrose yield, which are the two key elements for optimised sugar production. Chemical characteristics of the plants need to be taken into consideration as high sucrose is seldom found in fibrous varieties. Leaf sheath tightness is another characteristic that would go well with leaf sheath hairiness, because though not tested in this work-would make it difficult for the larvae to get to the smooth adaxial surface of the leaf. The hardness of trichomes is another feature that needs to be investigated, because a variety may have dense, but soft pubescence that does not repel even the most sensitive larvae, neonates. At present, integrating plant resistance with cultural control, i.e. field hygiene etc. is cost-beneficial for the sugar industry.<br>Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2002.

The use of entomopathogenic fungi to control agriculturally important pests, both in greenhouses and in the field, has been demonstrated by various authors for a number of years. This has been brought about by the development of resistance in certain pest species to chemical applications and a growing public awareness of the safety implications of residual insecticides. Several entomopathogenic fungi were tested against four insect pests found in the Republic of South Africa (RSA), the greenhouse whitefly, Trialeurodes vaporariorum, the rice weevil, Sitophilus oryzae, the grapevine mealybug, Planococcusficus and the sugarcane stem borer, Eldana saccharina. Further concentration, temperature and humidity studies were conducted with selected isolates on the rice weevil, S. oryzae. Sitophilus oryzae is considered one ofthe most important pests of stored grain. Several fungal isolates were tested against the rice weevil, four of which, B1, PPRI 6690, PPRI 6864 and PPRI 7067, were selected for further testing based on the mortality results over a 21 d period. Varying conidial concentrations were applied and at high doses of 1x10 -6 conidia ml -1 with mortality rates of to 84% achieved. LT 50 values ranged from 6 - 68d. Increased spore concentration resulted in an increase in overall mortality. Temperature and humidity was found to affect the infection potential of the four isolates tested. Four temperatures ranging from 15 - 30°C were tested. The highest mortality rates were obtained at 25°C where mortality ranged from 46 - 65% in 14d. Mortality rates decreased with decreasing temperature, and no mortality was recorded at 30°C. Temperature was found to significantly alter the LT 50 values, increasing the LT 50 with decreasing temperatures. Decreasing the humidity resulted in an increased LT 50 and a reduction in the overall mortality rates. The mortality of S. oryzae ranged according to the RH and isolate. Isolates Bland PPRI 6690 resulted in the highest mortalities of 80 and 83% at 92.5% RH, with LT 50's of 6.3d and 6.4d, respectively. Several entomopathogenic fungi were tested against T vaporariorum, P. ficus and E. saccharina, three key pests of South African crops. Nine fungal isolates were tested against the greenhouse whitefly, T vaporariorum, with mortalities ranging from 26.7 - 74.7% over 14d. Beauveria bassiana Isolates Bl and PPRl 6690 produced the highest mortality rates and were recommended for further pathogenicity testing against T. vaporariorum. Planococcus ficus is a common pest ofvineyards in the Western Cape Province, South Africa. Nine entomopathogenic fungi were screened against P.ficus, only two of which produced mortality. Eldana saccharina is a stalk borer, which infests sugarcane in large areas of Southern Africa. Five isolates were tested against second and third instar larvae, three of which, B1, PPRl 6864 and PPRl 6690 resulted in mortalities. Mean percentage mortality was low for all three isolates. From the study it was evident that two of the isolates tested, Bland PPRI 6690 (B. bassiana), showed potential against three of the four pests, and two isolates of Lecanicillium lecanii caused mortality in P. ficus. Further research and understanding of the effect of environmental conditions, spore concentration and epizootic potential would result in the further development of these isolates as future biological control agents.<br>Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

Organic insecticides play a big role in reducing the usage of chemical insecticides and their negative impact on the environment. Bacillus thuringiensis (Bt) spays are the only tool that organic farmers are allowed to use for the control of pests. Genetic engineering and modification of crops have been made possible with scientific advances in cell and molecular biology. These advances are used to transfer some of the Bt Cry toxins into crops for control of target species to reduce yield loss. Bt maize were commercialised for the first time in South Africa in 1998 and the economic important stem borers, Busseola fusca (Fuller) (Lepidoptera: Noctuidae), Chilo partellus (Swinhoe) (Lepidoptera: Crambidae) and Sesamia calamistis (Hampson) (Lepidoptera: Noctuidae) were exposed to the Cry1Ab toxin that is found in Bt maize. Busseola fusca developed resistance to Cry1Ab under field conditions within eight years after it had been released. Eldana saccharina (Walker) (Lepidoptera: Pyralidae) is a major pest on sugarcane in South Africa and although it has not been recorded on maize in this country, is it known as a major pest of maize in other African countries. African armyworm, Spodoptera exempta (Walker) (Lepidoptera: Noctuidae) has a very wide distribution in Africa and is known to be an occasional pest on maize. The aims of this dissertation were to determine the efficacy of Bt spray applications for control of four lepidopteran pests and whether development of Cry1Ab resistance by B. fusca caused a loss in susceptibility to other Bt toxins (i.e. cross-resistance). Susceptibility bioassays with 10 day old larvae were conducted under laboratory conditions. Treatments included application of various dosage rates of Dipel® and deltamethrin as well as exposure to MON810 (maize leaves). Stemborer populations of C. partellus, E. saccharina, and B. fusca (Venda) as well as the S. exempta were effectively controlled by the Bt spray, Dipel®. Care should be taken not be interpret the percentage C. partellus, E. saccharina and S. exempta larvae that survived after exposure to MON810 and Bt spray treatments as development of resistance without verification of these experiments with earlier instars that are known to be more susceptible. Spodoptera exempta is active throughout a year in temperate zones of Africa. If S. exempta develop resistance to Cry toxins and Bt maize events would be released for commercial planting in these areas, S. exempta pose a threat added to their injuriousness. Busseola fusca larvae were sampled from Venda (susceptible population), Ventersdorp and the Vaalharts Irrigation Scheme (resistant population). The Ventersdorp B. fusca population was controlled by MON810 and MON89034 and Bt sprays, but the percentage larvae that survived showed reduced susceptibility within the population. Dipel® treatments, MON810 and MON89034 did not provide effective control of the Vaalharts B. fusca population reported to be resistant to Cry1Ab, in two experiments. The high survival rates indicate a reduction in susceptibility to Cry toxins other than Cry1Ab and therefore development of cross resistance in the Vaalharts B. fusca population.<br>MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

The fungal disease smut (causal agent: Ustilago scitaminea H. & P. Sydow) and stalk borer Eldana saccharina Walker place major constraints on sugarcane agriculture in South Africa. The best approach for management is the introduction of resistant cultivars; however, conventional field-based screening for pest and disease resistance is a lengthy process. This study evaluated in vitro techniques combined with artificial inoculation of 12 week old in vitro plantlets and 8-10 week old embryogenic calli as rapid screening methods. Preliminary investigations were conducted on cultivars with known field ratings to smut and E. saccharina: NCo376, N26 and N39; and 5 „test‟ cultivars, whose identities were undisclosed until completion of experiments, were used to assess the accuracy of protocols. Infective U. scitaminea sporidia generated from teliospores, were used as inocula. Development of a callus protocol was unsuccessful due to sporidial and mycelial overgrowth, despite addition of a contact fungicide, Dithane M-45® (0.025 g/l) and a biocide/fungicide, PPMTM (5 ml/l), to media. Plantlet inoculation by injection, 1 cm above the apical meristem, resulted in 12% and 20% of smut susceptible NCo376 plantlets producing smut whips after 5 weeks when inoculated with 1 x 106 and 1 x 109 sporidia/ml, respectively. Smut whip production in 5 of the 8 (63%) cultivars inoculated with the lower sporidial concentration correlated with their field resistance ratings. In addition, whips harvested from in vitro plantlets were a valuable source of aseptic teliospores for future research. Ongoing work involves inoculation of NCo376 calli with such teliospores and maintenance on medium with PPMTM - emergence of whips from plantlets remains to be assessed. The E. saccharina screening protocol involved surface decontamination of eggs with 1% sodium hypochlorite (NaOCl) for 15 min. Feeding bioassays were conducted by placement of first instar larvae on in vitro plantlets and calli for 3 and 2 weeks, respectively. Larval mass, length and percentage infestation were recorded. Although greater larval size was expected in susceptible compared with resistant cultivars, the results did not support this. Significant differences in plantlet infestation were observed between susceptible (94-98%) and resistant (72-86%) lines. No significant differences were found in the callus feeding bioassay. However, a 24 h callus choice bioassay which investigated larval preference between callus genotypes compared with NCo376, showed significant differences and correctly discerned cultivar susceptibility according to field ratings.<br>Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2010.