Faculty of AgriSciences
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The Faculty of AgriSciences at Stellenbosch University (SU) is held in high esteem at national and international levels for the quality of its training and research and also as consultant in the agricultural and forestry industry.
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Browsing Faculty of AgriSciences by browse.metadata.advisor "Addison, M. F."
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- ItemAssessment of toxic baits for the control of ants (Hymenoptera : Formicidae) in South African vineyards(Stellenbosch : Stellenbosch University, 2008-12) Nyamukondiwa, Casper; Addison, Pia; Addison, M. F.; Stellenbosch University. Faculty of AgriSciences. Dept. of Conservation Ecology and Entomology.ENGLISH ABSTRACT: Ant infestations comprising the Argentine ant Linepithema humile (Mayr), common pugnacious ant Anoplolepis custodiens (F. Smith) and cocktail ant Crematogaster peringueyi Emery are a widespread pest problem in South African vineyards. Integrated Pest Management (IPM) programmes aimed at suppressing the problematic honeydew excreting vine mealybug Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae) on grapes must include ant control to optimize the effectiveness and efficacy of mealybug natural enemies. If ants are eliminated, natural enemies are able to contain mealybugs below the Economic Threshold Level (ETL). Current strategies for ant control are limited and generally include the application of long term residual insecticides that are detrimental to the environment, labour intensive to apply and can disrupt natural biological control if applied incorrectly. A more practical method of ant control using low toxicity baits was therefore investigated. Field bait preference and bait acceptance assessments aimed at determining bait repellency and palatability, respectively, were carried out during spring, summer and autumn in three vineyards of the Cape winelands region during 2007/08. Five toxicants comprising gourmet ant bait (0.5%), boric acid (0.5%), fipronil (0.0001%), fenoxycarb (0.5%) and spinosad (0.01%) dissolved in 25% sugar solution were tested against a 25% sucrose solution control. Gourmet ant bait was significantly more preferred and accepted by all ant species than the other baits. Laboratory bait efficacy assessments using four insecticides (gourmet, boric acid & spinosad) at concentrations of 0.25; 0.5; 1; 2 and 4 times the field dose and fipronil at 0.015625; 0.03125; 0.0625; 0.125; 0.25 times the field dose were carried out. Results revealed that boric acid (2%), gourmet ant bait (2%) and fipronil (1.0 X 10-5%) exhibited delayed toxicity for L. humile and C. peringueyi while spinosad (0.01%) showed delayed action on L. humile. Field foraging activity and food preference tests were also carried out for the three ant species during 2007/08. Foraging activity trials revealed that vineyard foraging activity of L. humile is higher relative to A. custodiens and C. peringueyi. This means fewer bait stations are required for effective L. humile control making low toxicity baits a more affordable and practical method of controlling L. humile than the other two ant species. Food preference trials showed that L. humile and C. peringueyi have a high preference for sugar while A. custodiens significantly preferred tuna over other baits. However, all ant species had a preference for wet baits (25% sugar water, 25% honey, tuna & agar) as opposed to dry ones (fish meal, sorghum grit, peanut butter & dog food). This research concludes that low toxicity baits show potential in ant pest management and can offer producers with a more practical, economical and environmentally friendly method of ant control which is compatible with vineyard IPM programmes.
- ItemBiological and ecological factors contributing to the successful use of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) for the control of codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae) under South African conditions(Stellenbosch : Stellenbosch University, 2011-12) De Waal, Jeanne Yvonne; Malan, Antoinette P.; Addison, M. F.; Addison, Pia; Stellenbosch University. Faculty of AgriSciences. Dept. of Conservation Ecology and Entomology.ENGLISH ABSTRACT: Codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae) is a devastating pest of pome fruit in temperate regions of the world. Control of this pest, previously involved the extensive use of broad-spectrum insecticides. However, concerns over human safety, environmental impact, widespread dispersal of resistant populations of codling moth and the sustainability of synthetic pesticides in agroecosystems, has encouraged the development and use of alternative environmentally-friendly pest management technologies including the use of entomopathogenic nematodes. These nematodes are lethal pathogens of insects and belong to the families Steinernematidae and Heterorhabditidae, and are ideal candidates for incorporation into the integrated pest management programme currently being developed for residue-free pome fruit production in South Africa. The biological and ecological factors pertaining to the successful use of these nematodes for the control of codling moth were investigated in this study. Their use for bin-disinfestations was evaluated, focusing on the optimum handling conditions to ensure the survival and subsequent efficacy of the nematodes. The study proved that the local isolate SF41 of Heterorhabditis zealandica Poinar 1990 could be used for successful bin-disinfestation. The use of the same nematode isolate was also investigated for the disinfestation of mulch layers of diapausing codling moth larvae. An insect containment device which allowed for direct trial efficacy evaluation was identified and ecological factors pertaining to the successful use of nematodes for mulch disinfestation were investigated. The biological control potential of local nematode isolates, which had previously never been tested against codling moth larvae, was investigated in the laboratory under conditions as can be expected during orchard applications. The efficacy of the selected isolates was confirmed in field experiments. Innovative insect containment methods for above-ground trial efficacy evaluation in the field were investigated. Desiccation proved to be the most limiting factor to the survival and subsequent efficacy of the nematodes during field applications in temperate regions. The effect of low moisture levels on H. zealandica’s efficacy to control diapausing codling moth larvae was subsequently investigated and a starch-based formulation was further tested to overcome the issue of desiccation. Conclusive results indicated that there were several biological and ecological factors influencing the survival of nematodes and illustrated how these factors could be manipulated to overcome these issues and thereby ensure the efficacy of treatments. This is the first report of its kind to comprehensively investigate the use of South African entomopathogenic nematodes for the control of diapausing codling moth larvae and all results emanating from the study can be integrated into a framework for the commercial use of these nematodes in this regard in future.
- ItemCharacterisation of nematode symbiotic bacteria and the in vitro liquid culture of Heterorhabditis zealandica and Steinernema yirgalemense(Stellenbosch : Stellenbosch University, 2013-03) Ferreira, Tiarin; Malan, Antoinette P.; Addison, Pia; Addison, M. F.; Stellenbosch University. Faculty of AgriSciences. Dept. of Conservation Ecology and Entomology.ENGLISH ABSTRACT: Entomopathogenic nematodes have the potential to be outstanding biocontrol agents against agricultural pest insects. Combined with their bacterial symbionts, these biocontrol agents have proven to be very effective against numerous pests. The nematodes belong to the families Steinernematidae and Heterorhabditidae, and are ideal to be used in, and integrated with, pest management systems. There is a dire need for new and innovative methods to control agricultural pests, as numerous pest insects have developed resistance against broad-spectrum insecticides. Together with the environmental impact of these insecticides and the safety aspect regarding humans and animals, the need to develop new technologies, including entomopathogenic nematodes for pest management, is high. In this study, the associated symbiotic bacteria of three entomopathogenic nematodes species were isolated, and the potential of two nematode species to be successfully mass cultured in liquid medium was evaluated. Regarding the symbiotic bacteria, results from the study showed that bacteria species from all three nematode species, Heterorhabditis noenieputensis, Steinernema khoisanae and Heterorhabditis zealandica, were novel. Heterorhabditis noenieputensis was isolated in the Mpumalanga province during a previous survey conducted in citrus orchards. The bacterium isolated from this nematode belongs to the genus Photorhabdus, and bear closest similarity (98.6%) to the type strain of P. luminescens subsp laumondii (TT01T). Photorhabdus luminescens subsp. noenieputensis subsp. nov., derives its name from the area where the nematode was sourced, namely the farm Springbokvlei, near the settlement Noenieput close to the Namibian border. Thus far, 85 Steinernema spp. have been described worldwide, including S. khoisanae which was isolated in the Western Cape province of South Africa. Four S. khoisanae strains, namely SF87, SF80, SF362 and 106-C, were used for characterisating the new bacteria from different localities in South Africa. Using the neighbor-joining method, all the strains were aligned with 97% homology to the 16S rRNA sequences of several Xenorhabdus- type strains, indicating that they belonged to the same genus. The multigene approach was used to distinguish between the Xenorhabdus spp. and partial recA, dnaN, gltX, gyrB and infB gene sequences of the various strains were analysed. The bacterium species was named Xenorhabdus khoisanae sp. nov. after the nematode from which it was isolated. The results showed that the third bacterium species, which was isolated from H. zealandica, was new. The sequence of the bacteria strain clustered with the type strains of P. temperata and P. asymbiotica, indicate that it belonged to the genus Photorhabdus. This is the first study to show that H. zealandica associates with a luminescent Photorhabdus species, rather than with the known non-luminescent P. temperata. The potential of H. zealandica and Steinernema yirgalemense mass culture in liquid was investigated. Results illustrated that H. zealandica and its P. luminescens symbiont can be successfully cultured in liquid. However, two generations occurred during the process time, instead of the desirable one-generation. The growth curve of the symbiotic bacteria during the process time was measured, in order to determine when the stationary phase was reached, with the results showing this to occur after 36 h. Therefore, the optimum amount of time required for inoculating the IJs and for aiding in maximum infective juvenile (IJ) recovery is 36 h for adding the nematodes post pre-culturing of the bacteria. Future research goals should be to increase the percentage recovery in liquid culture, which would increase the number of nematodes produced per ml, which would, therefore, reduce the processing time significantly. The results from mass culturing the second nematode species, S. yirgalemense, indicated an asynchronous nematode development in the first generation. Growth curves were performed with the symbiotic bacteria that showed the exponential phase of Xenorhabdus started after 15 h, and that, after 42 h, the stationary phase was reached, with an average of 51 × 107 cfu·ml-1. Bioassays were performed to compare the virulence between in vitro- and in vivo-produced nematodes, with the results showing that the in vitro-produced nematodes were significantly less virulent than were the nematodes produced in vivo. The success obtained with the production of S. yirgalemense in liquid culture can serve as the first step in the optimising and upscaling of the commercial production of nematodes in industrial fermenters. The last aim of the current study was to determine when Xenorhabdus reached the stationary phase, when it is grown in a 20-L fermenter, as this would be the optimum time at which to add the IJs of S. yirgalemense. Such characteristics as the effect of stationary phase conditions on the bacterial cell density and on the DO2 rate in the fermenter were investigated. The results showed that the stationary phase of Xenorhabdus was reached after 36 h at 30˚C, which took 6 h less than did the same procedures followed with the Xenorhabdus sp. cultured in Erlenmeyer flasks on orbital shakers. This is the first step toward the liquid mass culturing of S. yirgalemense in industrial-size fermenters. Data from this study indicated the optimum amount of time that is required for adding nematodes to the bacterial culture in the fermenter, and for ensuring the optimum recovery of IJs, as well as a subsequent high yield of nematodes within a minimum processing time. This is the first report of its kind to investigate comprehensively the successful liquid culture of two South African entomopathogenic nematode species for the sole purpose of evaluating potential commercialisation. Results emanating from this study could be used as groundwork in future, in combination with similar research such as culturing nematodes intensively in large fermenters.
- ItemEntomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) for the control of codling moth, Cydia pomonella (L.) under South African conditions(Stellenbosch : Stellenbosch University, 2008-12) De Waal, Jeanne Yvonne; Addison, Pia; Malan, Antoinette P.; Addison, M. F.; Stellenbosch University. Faculty of AgriSciences. Dept. of Conservation Ecology and Entomology.The codling moth, Cydia pomonella (L.), is a key pest in pome fruit orchards in South Africa. In the past, broad spectrum insecticides were predominantly used for the local control of this moth in orchards. Concerns over human safety, environmental impact, widespread dispersal of resistant populations of codling moth and sustainability of synthetic pesticide use have necessitated the development and use of alternative pest management technologies, products and programmes, such as the use of entomopathogenic nematodes (EPNs) for the control of codling moth. Entomopathogenic nematodes belonging to either Steinernematidae or Heterorhabditidae are ideal candidates for incorporation into the integrated pest management programme currently being developed for pome fruit orchards throughout South Africa with the ultimate aim of producing residuefree fruit. However, these lethal pathogens of insects are not exempted from governmental registration requirements and have therefore not yet been commercialized in South Africa. A nontarget survey was conducted to find suitable isolates of EPNs from local soils and to test their effectiveness as control agents against the codling moth. Soil samples were collected from several habitats and regions throughout South Africa and nematodes were recovered using the insect baiting technique. All EPN isolates obtained were identified to species level using a molecular taxonomic approach. Entomopathogenic nematodes were recovered from 20 of the 200 soil samples (10 %). Of these, eight (40 %) yielded Steinernema spp., identified as three isolates of S. khoisanae and five undescribed Steinernema spp. The other 12 (60 %) of the samples were positive for Heterorhabditis spp. and included six isolates of H. bacteriophora, five H. zealandica and one H. safricana. These isolates were then evaluated in laboratory bioassays for their potential as microbial agents of codling moth under varying conditions. A morphometric study indicated that all natural openings (mouth, anus and spiracles) of final instar codling moth were large enough for the infective juveniles (IJs) of all tested EPN species to gain entry into the larvae.
- ItemThe in vivo production of Heterorhabditis zealandica and Heterorhabditis bacteriophora(Stellenbosch : Stellenbosch University, 2012-03) Van Zyl, Carolina; Malan, Antoinette P.; Addison, Pia; Addison, M. F.; Stellenbosch University. Faculty of AgriSciences. Dept. of Conservation Ecology and Entomology.ENGLISH ABSTRACT: The agricultural industry in South Africa is dominated by the use of insecticides. Producers rely heavily on chemicals that cause increased risk to health, the environment and ecology, rapid resistance development in key insect pests and pesticide residues on crops. The increased concern regarding the impact of these pest management practices on the environment and alternative pest management strategies are being investigated. Entomopathogenic nematodes (EPNs) have been identified as being promising biological control agents of key insect pests. The two EPN genera that have shown promise for use as biological control agents within an integrated pest management programme areSteinernema and Heterorhabditis. Commercialisation and the successful use of EPNs to control pests in North America, Australia, Europe and Asia have confirmed the effectiveness of these organisms as biological control agents. Unfortunately, EPNs in large enough numbers for commercial field applications are not yet available on the South African market. Large numbers of EPNs can be produced through either in vivo or in vitro culturing practices. The objective of this study was to streamline the in vivo production process by using two endemic EPN species, Heterorhabditis zealandica (SF41) and H. bacteriophora (SF351). These EPN isolates have been shown to be effective control agents of codling moth Cydia pomonella, false codling moth Thaumatotibia leucotreta, obscure mealybug Pseudococcus viburni, and the banded fruit weevil Phlyctinus callosus. A comparative study was conducted to identify suitable host insects for EPN production of local H. zealandica (SF41) and H. bacteriophora (SF351) strains. Hosts were selected according to their susceptibility to the two EPN species used, their general availability and the ease and cost of rearing. Wax moth larvae Galleria mellonella (WML) and mealworms Tenebrio molitor (MW) were selected as hosts. In order to produce nematodes of consistent quality, a continuous source of host insects reared on a standardised diet was required. WML and MW were each reared on five different diets in the dark at ±26°C. A superior diet for each host was selected according to the diet that produced, on average, the larvae with the highest body mass within a specific timeframe. The heaviest WML, at an average weight of 0.19 g per larva, were produced on a diet consisting of 118 g wheat flour, 206 g wheat bran, 118 g milk powder, 88 g brewer‟s yeast, 24 g wax powder, 175 ml honey and 175 ml glycerol. The heaviest MW larvae weighed, on average, 0.0154 g per larva, and were produced on a diet consisting of 100% wheat bran. To confirm the hypothesis that a linear relationship exists between the weight of a host and the number of nematodes produced from that host, a study was conducted to determine the number of H. zealandica and H. bacteriophora produced per g of host. WML, MW, codling moth larvae and false codling moth larvae were weighed individually and inoculated with the two nematode species respectively. In addition, nematode production in frozen MW and WML was tested. The number of nematodes harvested from each host was counted, and the average number of nematode progeny produced in each host was calculated. A significant linear correlation between the weight of WML and MW and the number of H. zealandica and H. bacteriophora respectively produced confirmed the hypothesis that nematode production within the specified host increases with an increase in host weight. WML produced the highest number of H. zealandica and H. bacteriophora per g of host (1 459 205 ± 113 670 and 1 898 512 ± 94 355), followed by MW larvae (836 690 ± 121 252 and 414 566 ± 67 017). Lower numbers of H. zealandica and H. bacteriophora per g codling moth (57 582 ± 10 026 and 39 653 ± 8 276) and per g false codling moth (192 867 ± 13 488 and 97 652 ± 23 404) were produced. Successful infection of a suitable insect host is one of the key factors in an efficient in vivo nematode production process. Three inoculation techniques were compared using H. zealandica and H. bacteriophora: inoculation with a pipette; shaking of hosts in the nematode inoculum; and immersion of hosts in the nematode suspension. With each inoculation technique, WML and MW were used as host larvae and were inoculated with nematodes at a concentration of 200 infective juveniles (IJs) / larva. The percentage mortality of insect hosts was determined after two days, and EPN infectivity, confirmed by colour change and dissection, after seven days. The highest percentage EPN infection was obtained using pipetting for both nematode isolates and hosts. Nematode infection rates for all nematode-host combinations obtained with pipetting were above 90%, with the exception of MW inoculation with H. bacteriophora, where the percentage of infection obtained was 76%. The current study conclusively demonstrated that variations in infection levels occur, depending on the inoculation technique used. In an additional effort to enhance infectivity during inoculation, H. zealandica, H. bacteriophora and MW were subjected to host-stressor regimes and to nematode- infectivity-enhancing additives. Three treatments, plus a control treatment, were compared. Exposing MW to 70°C tap water prior to inoculation did not increase infection levels. On the contrary, reduced infection levels were observed with host immersion in 70°C tap water followed by inoculation with H. bacteriophora, compared to the control. Only 12% infection was obtained compared to the 48% infection achieved in the control. Infection obtained using H. zealandica was 21%. Treating H. zealandica and H. bacteriophora IJs withMn2+SO4.H20 in a suspension, prior to inoculating MW, did not significantly enhance nematode virulence. Inoculation of MW with treated H. zealandica IJs led to an infection rate of 81%, compared to the control, with which 80% infection rate was obtained. Heterorhabditis bacteriophora caused 47% MW infection, compared to the control, which was subject to 48% infection. A combination of the two above-mentioned treatments did not enhance the infection levels either. Immersing MW into 70°C tap water prior to inoculation with nematodes treated with Mn2+SO4.H20 led to infection levels of 13% and 9% respectively when H. bacteriophora and H. zealandica were used. Future research is required to optimise the protocol used in this study of subjecting MW and local nematode isolates to stressor regimes. The ability of two formulations to maintain biological activity and virulence of H. zealandica was investigated. A quality standard control measure was used to measure the percentage survival and virulence of formulated H. zealandica over a period of 21 days. IJs were formulated into Pesta granules and coconut fibres, while nematodes stored in tap water served as the control. The numbers of live H. zealandica in Pesta granules and coconut fibres decreased drastically after seven days of storage. The survival of nematodes in Pesta granules dropped to 9.79% after 21 days compared to the control, where the survival rate was 79.79%. Nematode survival in coconut fibres was even lower, at 25.84% after seven days and 2.25% after 21 days. After 21 days in storage, 100%+of nematodes survived in the control for coconut fibres. The application of the standard quality control measure, which was used to determine the virulence of formulated H. zealandica, proved to be ineffective. Higher MW mortality rates were obtained in the control where no nematodes were added to larvae, compared to where nematodes were added in varying dosages. However, adjusting certain aspects in the protocol of this quality control measure specifically to accommodate local conditions could possibly make it a more effective tool for measuring endemic nematode virulence.
- ItemOptimization of a mass-rearing system to produce codling moth, Cydia pomonella, for a Sterile Insect Release programme in South Africa(Stellenbosch : University of Stellenbosch, 2011-03) Stenekamp, Daleen; Addison, Pia; Addison, M. F.; University of Stellenbosch. Faculty of AgriSciences. Dept. of Conservation Ecology and Entomology.ENGLISH ABSTRACT: Codling moth, Cydia pomonella, is a worldwide pest and of major economic importance to the South African pome fruit industry. Sterile insect release is applied as a component of area-wide integrated pest management and includes the mass-rearing, sterilization and the release of the sterile insects. For sterile insect release, the improvements of rearing methods in terms of the quality of the diet ingredients and the economical aspect of the rearing method are examined. The effect of genetically modified maize meal, containing the Bacillus thuringiensis gene, in an artificial medium for codling moth rearing, is determined. The use of even a small amount of Bacillus thuringiensis resulted in larval mortality and prolonged development. These results are detrimental to a mass-rearing facility and must be considered by any rearing facility that uses genetically modified maize meal if the insect is sensitive to the gene. An alternative to maize meal in the artificial medium was tested and whole wheat flour was considered to be a suitable replacement. Agar agar is an expensive gelling agent used in the artificial medium. An alternative for agar agar (Kelcogel, Elastigel and carrageenen) is tested and the biological effect on codling moth is determined. Factors such as mortality, pupal and moth weight, longevity, fecundity and development time were used as quality parameters. Results showed that Elastigel was a suitable replacement for agar agar, with bigger pupae and moths, higher fecundity and increased longevity. The economical advantage of the replacement is a 40.91% reduction of the diet cost. The other gelling agents tested also gave acceptable results and can be considered if shortages of agar agar or Elastigel occur. A new method of mass-rearing codling moth larvae in a closed rearing system using large trays placed in a ventilated box is designed. This method is more cost and space effective as a smaller area is needed to rear a large number of moths. The risk of diet contamination is less because of the closed environment and more economical and effective air handling. This is the first report of its kind to describe the mass-rearing of codling moth in a closed environment and the risks involved in using genetically modified maize meal in an artificial diet for the codling moth. These results should be incorporated into existing mass-rearing facilities or taking into consideration when designing new mass-rearing facilities.