The feasibility of managing codling moth, Cydia pomonella (L.), with mixtures of Bacillus thuringiensis-based microbial and selected chemical insecticides was studied under laboratory and field conditions. Joint actions (synergism, additivity and antagonism) of these mixtures were determined for both egg and larval stages. Higher rates of microbial control agents (MCAs), Dipel�� 2X and MVP��, combined with low to medium rates of chemical insecticides gave additive or synergistic egg and/or larval mortality. Esfenvalerate and permethrin synergised the effect of these MCAs, whereas carbaryl, azinphosmethyl, phosmet and diflubenzuron had an additive effects. For all chemical insecticides, often the interactions were better with Dipel than MVP. Generally, egg exposure to low rates of the chemical insecticides alone or in combination with MCAs, caused the larvae hatching from these eggs to be more susceptible to insecticides-MCA mixtures. Most mixtures produced better results when treated at the blackhead stage of egg development. Adult and larval exposure to sublethal rates of azinphosmethyl, phosmet, carbaryl, esfenvalerate and permethrin manifested short- and long-term effects. Adult exposure caused deleterious effects on the survival and oviposition, and the larval exposures affected survival and pupal formation. Carbaryl, esfenvalerate and permethrin affected the pupal survival and subsequent eclosion of adults. Only esfenvalerate and permethrin significantly extended the larval period and reduced pupal weight and subsequent egg deposition. The deleterious sublethal effects of pyrethroids observed in the present study may provide added control of codling moth in the field. Diflubenzuron and MCAs had neither adulticidal activity nor long-term effects on codling moth biology. Results of field efficacy tests indicate that utilization of mixtures of Dipel with low rates of pyrethroids (0.1-0.2X) or diflubenzuron (0.25-0.50X) in the codling moth management is possible and indeed might be beneficial. Dipel-diflubenzuron mixtures were only suitable when applied 3 times/generation. These mixtures were environmentally safe and did not cause any outbreak of phytophagous mites. Standard application rate (2 times/generation) and timing were found to be appropriate for Dipel-pyrethroid (esfenvalerate and permethrin) mixtures. These pyrethroids at all test rates (0.1-1.0X), applied alone or mixed with MCAs, were toxic to predatory phytoseiids, thus caused an outbreak of European red mite and yellow spider mites particularly at higher rates. However, mixtures of Dipel and 0.1X of esfenvalerate/permethrin were less disruptive than corresponding 0.2-1.0X standard rate. These mixtures were selective to most predatory insects, spiders and Zetzellia malt. Hence, use of these mixtures instead of full rate of pyrethroids may improve the prospects of biological control of insect and mite pests in apple orchards. / Graduation date: 1996

Thesis (PhD (Conservation Ecology and Entomology))--University of Stellenbosch, 2011. / 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. / AFRIKAANSE OPSOMMING: Die kodlingmot, Cydia pomonella, is van ekonomiese belang vir die Suid-Afrikaanse kernvrugte bedryf. Die steriele insek tegniek word gebruik as ‘n komponent in area-wye geïntegreerde plaagbeheer en sluit in die massa-aanteel, sterilisering en vrylaat van steriele insekte. Vir die steriele insek tegniek is die verbetering van die massa-aanteel van die kodlingmot in terme van kwaliteit van die dieet en die ekonomiese aspek van die aanteel metode ondersoek. Die effek van genetiese gemanipuleerde mieliemeel wat die Bacillus thuringiensis geen bevat, in ‘n kunsmatige voedselmedium vir die aanteel van kodlingmot, is bepaal. Daar is gevind dat die gebruik van selfs ‘n klein persentasie Bacillus thuringiensis in die mieliemeel, mortaliteit en ‘n verlengde lewenssiklus in kodlingmot veroorsaak. Die gevolge is nadelig vir ‘n massa-aanteel fasiliteit en behoort in ag geneem te word vir enige insek wat op ‘n kunsmatige medium, wat mieliemeel bevat, geteel word, mits die insek sensitief is vir Bacillus thuringiensis. ‘n Alternatiewe bestanddeel vir mieliemeel, volkoringmeel, word aanbeveel. Agar agar is ‘n duur verdikkingsagent wat in kunsmatige mediums gebruik word. ‘n Alternatief vir agar agar (Kelcogel, Elastigel en carrageenen) is getoets en die biologiese effek op die kodlingmot is bepaal. Faktore soos mortaliteit, papie en mot gewig, langlewendheid, vrugbaarheid en lengte van lewenssiklus was gebruik as kwaliteit parameters. Resultate het getoon dat Elastigel ‘n geskikte plaasvevanger is van agar agar, met groter papies en motte, groter vrugbaarheid en langlewendheid. Die ekonomiese gevolg van die plaasvervanger, is ‘n vermindering van 40.91% van die dieetkoste. Die ander verdikkingagente wat is getoets is, het aanvaarbare resultate gelewer wat noodsaaklik is indien daar ‘n tekort van Elastigel of agar agar ontwikkel. ‘n Nuwe metode van massa-aanteel van kodlingmot larwes is bepaal. Die metode behels ‘n geslote sisteem, waar groter aanteel bakke in ‘n geslote, geventileerde boks geplaas word. Die metode is koste en spasie effektief en ‘n kleiner area word benodig om ‘n groter aantal motte te lewer. Die risiko van kontaminasie van die dieet word verminder as gevolg van die geslote sisteem wat gebruik word en meer ekonomiese en effektiewe lugversorging word gebruik. Hierdie is die eerste verslag van sy soort wat die massa-aanteel van kodlingmot in ‘n geslote sisteem beskryf en wat die risiko aandui van geneties gemanipuleerde mieliemeel in ‘n kunsmatige medium vir die kodlingmot. Hierdie resultate behoort in ag geneem te word vir reeds bestaande massa-aanteel fasiliteite of met die ontwerp van nuwe massa-aanteel fasiliteite.

Cydia pomonella (Linnaeus) (Family: Tortricidae), the codling moth, is considered one of the most significant pests of apples and pears worldwide, causing up to 80% crop loss in orchards if no control measures are applied. Cydia pomonella is oligophagous feeding on a number of alternate hosts including quince, walnuts, apricots, peaches, plums and nectarines. Historically the control of this pest has been achieved with the use of various chemical control strategies which have maintained pest levels below the economic threshold at a relatively low cost to the grower. However, there are serious concerns surrounding the use of chemical insecticides including the development of resistance in insect populations, the banning of various insecticides, regulations for lowering of the maximum residue level and employee and consumer safety. For this reason, alternate measures of control are slowly being adopted by growers such as mating disruption, cultural methods and the use of baculovirus biopesticides as part of integrated pest management programmes. The reluctance of growers to accept baculovirus or other biological control products in the past has been due to questionable product quality and inconsistencies in their field performance. Moreover, the development and application of biological control products is more costly than the use of chemical alternatives. Baculoviruses are arthropod specific viruses that are highly virulent to a number of lepidopteran species. Due to the virulence and host specificity of baculoviruses, Cydia pomonella granulovirus has been extensively and successfully used as part of integrated pest management systems for the control of C. pomonella in Europe and around the world, including South Africa. Commercial formulations have been typically based on the Mexican strain of CpGV. However due to long-term multiple applications of CpGV and the reliance on CpGV in organic farming practices in Europe, resistance to the CpGV-M strain has developed in a number of field populations of C. pomonella. This study aimed to identify and characterize novel isolates of CpGV in South Africa and compare their virulence with the commercial standard CpGV-M. Secondly, since C. pomonella is difficult to culture on a large scale, an alternate method of CpGV production was investigated in order to determine if CpGV could be produced more efficiently and at a reduced cost without negatively impacting the quality of the product. Several isolates of CpGV were recovered either from field collected larvae or from a laboratory-reared C. pomonella colony. Characterisation of DNA profiles using a variety of restriction enzymes revealed that only a single isolate, CpGV-SA, was genetically different from the Mexican strain of the virus used in the commercially available CpGV based products in South Africa. In dose-response bioassays using CpGV-SA, LC₅₀ and LC₉₀ values for neonate C. pomonella larvae were 3.18 x 10³ OBs/ml and 7.33 x 10⁴ respectively. A comparison of these values with those of CpGV-M indicated no significant difference in the virulence of the two isolates under laboratory conditions. This is a first report of a genetically distinct CpGV isolate in South Africa. The biological activity and novelty of CpGV-SA makes this isolate a potentially important tool for CpGV resistance management in South Africa. In order to justify production of CpGV in an alternative host, studies on the comparative biological performance of C. pomonella and T. leucotreta based on oviposition, time to hatch, larval developmental times and rearing efficiency as well as production costs were performed. Thaumatotibia leucotreta was found to be more fecund and to have significantly shorter egg and larval developmental times. In addition, larval production per unit of artificial diet was significantly higher than for C. pomonella. This resulted in T. leucotreta being more cost effective to produce with implications for reduced insectary space, sanitation practices as well as the labour component of production. Virus yield data generated by inoculation both C. pomonella and T. leucotreta with nine concentrations of CpGV resulted in comparable virus yields, justifying the continuation of the research into production of CpGV in T. leucotreta. It was important to determine the LC and LT values required for mass production of CpGV in late instar T. leucotreta larvae. Dose- and time-response bioassays with CpGV-M were conducted on artificial diet to determine these values. Fourth instar LC₅₀ and LC₉₀ values were 5.96 x 10³ OBs/ml and 1.64 x 10⁵ OBs/ml respectively. LT50 and LT90 values were 81.10 hours and 88.58 hours respectively. Fifth instar LC₅₀ and LC₉₀ values were 6.88 x 10⁴ OBs/ml and 9.78 x 10⁶ OBs/ml respectively. LT₅₀ and LT₉₀ values were 111.56 hours and 137.57 hours respectively. Virus produced in fourth instar T. leucotreta larvae was bioassayed against C. pomonella neonate larvae and compared to CpGV-M to establish if production in the heterologous host negatively affected the virulence of the isolate. No significant difference in virulence was observed between virus produced in T. leucotreta and that produced in C. pomonella. The data generated in the bioassays was used in CpGV mass production trials to evaluate production. All production methods tested produced acceptable virus yields. To examine the quality of the virus product, genomic DNA was extracted from larval cadavers and subjected to REN analysis with HindIII. The resulting DNA profiles indicated that the virus product was contaminated with the homologous virus, CrleGV. Based on the above results, the use of T. leucotreta as an alternate host for the in vivo production of CpGV on a commercial basis is not at this stage viable and requires further investigation before this production methodology can be reliable used to produce CpGV. However, this study has shown that CpGV can be produced in a homologous host, T. leucotreta and significant strides have been made towards developing a set of quality control standards that are essential for further development of successful production methodology. Finally a novel isolate of CpGV has been identified with comparable virulence to the CpGV-M. This is an important finding as it has broad reaching implications for resistance management of CpGV products in South Africa.