The false codling moth (FCM), Cryptophlehia Leucatreta, causes widespread damage to economically important fruit crops throughout sub-Saharan Africa. Fruit are rendered unfit for consumption once they have been stung by FCM larvae. Larval infestation of fruit can lead to significant pre-harvest losses or post-harvest waste, posing a major problem to the citrus industry. Current control of the pest includes the use of chemical pesticides. The larval form of FCM is known to be infected by a granulovirus called Cryptophlebia leucotreta granulovirus (CIGV). Granuloviruses are highly specific against their hosts and are harmless to vertebrates, plants and the environment. The development of CIGV into a biological control agent would offer an attractive and safer alternative for the control of this pest. A full characterisation of CIGV is required prior to the virus being disseminated into the environment. In this project, the characteristics of CIGV will be examined. Viral DNA was extracted from infected larvae and the DNA analysed by restriction fragment length polymorphism (RFLP). Fragmentation profiles of the South African and Cape Verde (CV3) isolates of the virus were compared, revealing distinct differences between them. The size of the CIGV-SA genome was calculated to be 112 kbp, identical to the size of the CV3 isolate. Physical maps for five restriction enzymes were constructed for the CIGV-SA genome. The alignment of these maps with maps the CV3 isolate (for the same enzymes) further highlighted the differences between the isolates. The genetic engineering of granuloviruses could significantly improve the speed of kill of these viruses. Therefore essential genes like egt and granulin were isolated (by PCR) and their position located in the genome. Both genes were sequenced and their phylogeny with other granulin and egt genes investigated. Finally, tbe incidence of CIGV in natural populations of FCM larvae was investigated, by screening field-collected larvae for the presence of the virus. CIGV was successfully detected from dot blots of larval DNA using both radiolabelled and non-radiolabelled probes and by PCR. Trends regarding the incidence of CIGV in natural populations of larvae were also determined.

The false codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyrick) (Lepidoptera: Tortricidae) is a serious pest of citrus and other crops in Sub-Saharan Africa. The introduction of the Cryptophlebia leucotreta granulovirus (CrleGV-SA) Cryptogran and Cryptex (biopesticides) has proven to be very effective in the control of FCM. However, markedly lower susceptibility of some codling moth (CM), Cydia pomonella (L.) populations to Cydia pomonella granulovirus (CpGV-M), another granulovirus product used in the control of CM’s in Europe have been reported. Genetic differences between FCM populations in South Africa have also been established. It is therefore possible that differences in the susceptibility of these geographically distinct FCM populations to CrleGV-SA might also exist. To investigate this phenomenon, a benchmark for pathogenecity was established. In continuation of previous work with Cryptogran against the 1st and 5th instar FCM larvae, dose-response relationships were established for all five larval instars of FCM. In surface dose-response bioassays, the LC50 values for the 2nd, 3rd and 4th instars were calculated to be 4.516 x 104, 1.662 x 105 and 2.205 x 106 occlusion bodies (OBs)/ml, respectively. The LC90 values for the 2nd, 3rd and 4th instars were calculated to be 4.287 x 106, 9.992 x 106 and 1.661 x 108 OBs/ml, respectively. Susceptibility to CrleGV-SA was found to decline with larval stage and increase with time of exposure. The protocol was used in guiding bioassays with field collected FCM larvae. Laboratory assays conducted with Cryptogran (at 1.661 x 108 OBs/ml) against field collected FCM larvae from Addo, Kirkwood, Citrusdal and Clanwilliam as well as a standard laboratory colony, showed a significant difference in pathogenecity in only one case. This significant difference was observed between 5th instars from the Addo colony and 5th instars from the other populations. Four geographically distinct FCM colonies from Addo, Citrusdal, Marble Hall and Nelspruit were also established. Since Cryptogran and Cryptex are always targeted against 1st instar FCM larvae in the field, further comparative laboratory assays were conducted with the Addo colony and an old laboratory colony. Cryptogran was significantly more pathogenic than Cryptex against both the Addo and the old colony. However, a high level of heterogeneity was observed in responses within each population.

Thesis (PhD )--Stellenbosch University, 1981. / EKSERP: Vanwee die voortdurende probleme wat die valskodlingmot, Cryptoph lebia leucotreta (Meyr.), by die verbouing van nawels in die Oos-Transvaalse Laeveld daarstel, is 'n indringende studie van hierdie insekprobleem onderneem. Die biologie van die insek is onder sowel laboratorium as natuurlike toestande nagevors. Daar is bevind dat temperatuur 'n belangrike rol in die ontwikkeling van die insek speel. By 20 tot 30°C het temperatuurstygings die ontwikkeling van die insek laat versnel. By 35°C het die lewensduur van die volwasse motte afgeneem en is geen eiers gelê nie . Die valskodlingmot is deur middle van 'n kunsmatige metode geteel om genoegsame proefmateriaal te verkry. Verskeie verbeteringe is in die bestaande tegniek aangebring.

Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is one of the major citrus pests of economic importance for South Africa’s citrus industry. It is endemic to Africa, and therefore a phytosanitary pest with zero tolerance by most export markets. The cryptic nature of T. leucotreta makes visual inspection an inefficient method for detecting neonate larvae in fruit in the packhouse. Therefore, a more accurate method for sorting infested fruit at the packhouse, particularly for newly infested fruit could ensure market access. A recent study showed that fruit infested by T. leucotreta emit a chemical profile different from that of a healthy fruit. Several studies provide evidence that parasitoids locate their hosts feeding on fruit by exploiting the novel chemical profiles produced due to host herbivory. The aim of this study was to evaluate the potential of using the naturally occurring behaviour of a larval parasitoid Agathis bishopi (Nixon) (Hymenoptera: Braconidae) for detection of T. leucotreta infested fruit, by determining which compound in infested fruit is attractive to parasitoids. Ytube olfactometer and flight-tunnel bioassays with healthy and T. leucotreta infested fruit showed a significantly stronger response of A. bishopi female parasitoids to infested fruit. Among the volatile compounds associated with T. leucotreta infested fruit, D-limonene elicited the strongest attraction to A. bishopi female parasitoids. Attraction of mated A. bishopi female parasitoids to T. leucotreta infested fruit and D-limonene significantly increased after oviposition experience. Behavioural responses of A. bishopi female parasitoids that were associated with T. leucotreta infested fruit were investigated to determine which behaviours are distinct and interpretable. Probing and oviposition behaviours were the most noticeable and were only elicited on infested fruit when parasitoids contacted T. leucotreta frass, indicating that chemical compounds in frass are short-range cues used for final host location. Since production of D-limonene by fruit is elevated due to herbivory by different pests including mechanical injury on fruit, response of A. bishopi female parasitoids to compounds in frass offers a more specific and potentially useful mechanism for development of a detection system for T. leucotreta infested fruit. Chemical analysis of T. leucotreta frass and conditioning A. bishopi parasitoids to respond behaviourally to compounds in frass is proposed.

Control of the citrus pest, false codling moth (FCM), Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae) is crucial for the South African citrus industry. The economic losses and phytosanitary status of this pest, coupled with increased consumer awareness and demands, has created a need for effective, IPM-compatible control measures for use against the soil-dwelling life stages of FCM. Promising developments in the field of microbial control through the use of entomopathogenic fungi (EPF) and entomopathogenic nematodes (EPNs) have highlighted the need for research regarding pupation biology, behaviour and survival of FCM, as a good understanding of biology of the target organism is an important component of any biological control programme. The aim of this study was to improve the current understanding of FCM pupation habits through the manipulation of soil texture class, ground cover, shading, soil compaction, air temperature, and soil moisture in the laboratory. These findings would then be used to aid the biological control programmes using EPF and EPNs against FCM in the soil. Three soil texture classes (sandy loam, silt loam and silty clay loam) were obtained from orchards for use in the study. FCM larvae were allowed to drop into the soil of their own accord and the pupation behaviour that followed was then captured on film with pupae formed in the soil being kept in order to measure adult eclosion. In general, very few abiotic factors had a clear influence on FCM pupation. Larval wandering time and distance was short, but also variable between individuals. Distance did increase when soils were moist. Pupation depth was shallow, with pupal cocoons generally being formed on the soil surface. Depth of pupation was less than one centimetre for all abiotic conditions, with little burrowing into soil. Eclosion success was higher for sandier soils when these were dry and uncompacted, but the addition of both moisture and soil compaction increased FCM eclosion success. FCM was sensitive to desiccation when the soils were dry and temperature limits of 15 °C and 32 °C had a strongly negative impact on eclosion success. Preferences for particular abiotic conditions were limited to only certain moisture conditions when interacting with soil texture class and a preference for pupating in soil when it is available. Limited preference was found for particular soil textures despite this having a strong influence on eclosion success, but individuals did appear to pupate in close proximity to one another. Viable direct habitat manipulation for FCM control could not be identified. These results and all of the abiotic variables measured have important implications for EPF and EPN application, survival and persistence in the soil in order to improve the ability of these biological control agents to control FCM. These are discussed in each chapter.

Thesis (MSc)--University of Stellenbosch, 2000. / ENGLISH ABSTRACT: The development time, survival and fecundity of Trichogrammatoidea cryptophlebiae Nagaraj a, an egg parasitoid of false codling moth, Cryptophlebia leucotreta Meyr., on . citrus were determined at 15,20,25 and 30°C. Using these values the intrinsic rate of increase (rm), mean generation time (T) and net replacement rate (Ra) were estimated at the respective temperatures. In addition, the data were used to estimate the minimum temperature required for development. The rmincreased from 15 (0,042) to 20°C (0,088) and then gradually decreased from 25 (0,069) to 30°C (0,012). The Ra was highest at 20°C (3,116 females/female), followed by 15°C (2,607 females/female), 25°C (1,894 females/female) and 30°C (1,092 females/female). T decreased as temperature increased from 7 days at 30°C to 22,79 days at 15°C. The estimated minimum temperature for development was 7,65°C which implies that this wasp should be able to survive and reproduce under normal winter conditions in the Western Cape. / AFRIKAANSE OPSOMMING: Die ontwikkelingstyd, oorlewing en vrugbaarheid van Trichogrammatoidea cryptophlebiae Nagaraj a, 'n eierparasitoied van valskodlingmot, Cryptophlebia leucotreta Meyr., op sitrus is bepaal by 15, 20, 25 en 30oe. Hierdie waardes is gebruik om die intrinsieke tempo van vermeerdering (rm), die gemiddelde generasietyd (T) en die netto vervangingstempo (Ro) te bepaal by die betrokke temperature. Die data is verder gebruik om die minimum temperatuur vir ontwikkeling te bepaal. Die rm waarde het toegeneem van 15°e (0,042) tot by 200e (0,088) en afgeneem van 25°e (0,069) tot 300e (0,012). Die Ro was die hoogste by 200e (3,116 wyfies/wyfie), gevolg deur 15°e (2,607 wyfies/wyfie), 25°e (1,894 wyfies/wyfie) en 300e (1,092 wyfies/wyfie). T het afgeneem met 'n toename in temperatuur en was 7 dae by 300e en 22,79 dae by 15°e. Die beraamde minimum temperatuur vir ontwikkeling was 7,65°e wat aandui dat hierdie wespe kan oorleef en voortplant by normale wintertoestande in die Wes-Kaap.

False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. Major concerns such as progressive insecticidal resistance, the negative impact of insecticides on the environment, as well as the influence of consumers opposed to chemical residues on fruit, created opportunities for biological control methods such as Sterile Insect Technology (SIT). This technology is now established in the Western and Eastern Cape provinces of South Africa as an effective, sustainable alternative to conventional FCM control methods. Due to the prevalence of the pest in all citrus producing areas of South Africa, potential for SIT to expand is enormous. Success of an SIT programme is highly dependent on efficient application of the technology to achieve its objectives in a timeous manner. The aim of this study was to advance the application of SIT for control of FCM on citrus in South Africa, by investigating the effect of certain critical stages in the process. The effect of long-distance transportation on fitness of irradiated FCM was determined, showing reduced performance with cold-immobilized transport. A significant decrease in flight ability and longevity of irradiated FCM was found, although critically, realized fecundity was not affected. The effect of two different insecticides in the pyrethroid and organophosphate chemical groups were investigated for their residual effect on mortality of released irradiated FCM, to determine if these pest control programmes could be integrated. Both chlorpyrifos and tau-fluvalinate were effective in killing irradiated FCM for a number of days after application, after which degradation of the active ingredient rendered it harmless. This effect was found to be similar for irradiated and non-irradiated males, consequently ratios of sterile : wild male FCM should be retained regardless of whether sprays are applied or not. The modes for release of sterile FCM in an SIT programme were investigated. Efficacy of ground and aerial release platforms were tested by evaluating the recovery of released irradiated male FCM in these orchards. More irradiated FCM were recovered in orchards released from the ground compared to air. However, an economic analysis of both methods shows application of irradiated insects over a large geographical area is more cost-effective by air. Depending on the terrain and size of the target area, a combination of both methods is ideal for application of SIT for control of FCM in citrus. Development of application technology for advance of the programme is discussed and recommendations for future research and development are offered.

The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), is an important pest of citrus in South Africa and challenging to manage due to its inconspicuous nature. An effective method currently being employed for the area-wide suppression of the FCM is the Sterile Insect Technique (SIT) and the effective dispersal of sterile moths is very important for success with SIT. This study was conducted in the Addo area of the Sundays River Valley (Eastern Cape) where the programme is commercially used. In this study, sterile male moths were released in different orchards on a citrus farm, and in nearby veld at different times of the year, and their dispersal was monitored through the use of pheromone traps. Various climatic factors were monitored. This provided insight into the local dispersal of sterile male FCM adults in response to abiotic cues (particularly climatic factors). The movement of the FCM in four citrus cultivars, namely lemons, navel and Valencia oranges and mandarins and in the nearby veld (open field), was determined at six different stages of the year. Results clearly indicated that sterile FCM movement is concentrated within citrus orchards, as very few moths were trapped beyond 30 m from the release point, particularly in navel and Valencia orchards. Of the climatic factors measured, minimum and maximum temperatures had the most significant influence on FCM dispersal, and based on the results, various recommendations are made for the releases of sterile FCM in an area-wide SIT management programmes on citrus. A better understanding of the dispersal capabilities of the FCM in an agricultural system, under different conditions and at different times of the year, is invaluable not only in improving release strategies in an SIT programme but in planning future control strategies against the FCM.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Thaumatotibia leucotreta, the false codling moth (FCM), is a phytosanitary pest in South Africa posing a substantial threat to many of the country’s international export markets. Its pest status is of high importance because it has a wide ecological range and has been reported in all areas where citrus is produced in South Africa. Many methods of control have been implemented, such as chemical and cultural control, mating disruption and sterile insect releases. There was a need to obtain a more accurate understanding of FCM biology on deciduous fruit in South Africa and this then us to pose the questions described in the chapters to follow. The first aim was focused on the possibility of FCM diapause during winter. If FCM were to undergo diapause this could pose further problems for control methods, but knowledge thereof could also assist in more accurate and timely control methods. Considering past research on other Lepidoptera species, four physiological traits were chosen as indicative of a diapause state. Water loss rate, metabolic rate and the supercooling points should be lower if the individuals were in a diapause state, with a higher fat content expected for these individuals. Diapause induction was attempted through a gradual lowering of the environmental temperature in combination with longer nights to simulate overwintering conditions. Diapause was not observed in these experimental individuals. The second aim was to better understand the field biology of FCM. This was studied through in-field flight ability studies and damage assessments on four fruit kinds. Six release dates were used to measure the flight ability. The highest recapture rates were at minimum temperatures above 16°C and maximum temperatures averaging above 30°C, although the recapture rates were not significant in relation to the amount released. The recapture rates in the different fruit kinds were not significantly different, with the amount recaptured at the closest distance of 30 m being significantly more than that of the other distances. This was also only for the last release at the warmest temperatures. Fruit damage assessments were conducted and we were able to rear wild FCM from Granny smith apples, Forelle pears, Larry Ann plums and Satsuma and Clementine citrus cultivars. Citrus infestations had the highest count and a prolonged occurrence compared to the other varieties, due to its later harvest period. The third aim was to study the developmental parameters of FCM in different fruit kinds and an artificial medium. Firstly, FCM did not infest apples, Royal Gala and Pink lady’s, under laboratory conditions. Results were obtained using Forelle pears, Clementines and Thompson seedless grapes. On average the grapes had the shortest FCM developmental time from egg to adult stage, followed by oranges and then pears. Pears had the lowest developmental success rate, with that of oranges and grapes being much higher. Infestations took place at the stalk end of the fruit for the grapes and oranges, with the pears being infested at the calyx end. Future research should include an in-field life cycle, to determine the life cycle of FCM on different economically important fruit kinds under field conditions. The focus could also be shifted to where FCM overwinter, leading to better preventative control leading to lower infestation pressure during harvest periods. This is of utmost importance in an environment where maximum residue levels for pesticides dictate market access. / AFRIKAANSE OPSOMMING: Thaumatotibia leucotreta, die vals kodling mot (VKM) is ‘n fitosanitere pes in Suid Afrika, wat kan lei tot groot finansiele verliese. Die VKM se wye gasheerreeks en die feit dat dit al in al die sitrus verbouings-areas in Suid Afrika opgelet is, maak dit ‘n ernstige pes. Daar word van verskeie beheer metodes gebruik gemaak, insluitend chemiese en kulturele metodes. In sommige areas word daar ook van paaringsontwrigting en steriele insek vrylatings gebruik gemaak en hierdie metodes word gewoonlik met ander gekombineer. Daar is ‘n groot behoefte vir meer inligting omtrent die status van VKM in sagtevrugte in Suid Afrika en het gelei tot die vrae wat in hierdie studie aangespreuk word. Die eerste doelwit was om te bepaal of die VKM wel diapouse ondergaan. Dit sal verskeie beheermetodes belemmer, maar kennis hiervan kan meer gefokusde en gevolglik meer effektiewe beheermaatreels tot gevolg hê. Daar is gekyk na vier fisiologiese eienskappe wat beduidend tot diapouse van ander Lepidoptera spesies is. Daar word verwag dat VKM wat diapouse ondervind ‘n hoër vetinhoud sal he, terwyl die metabolise tempo, “supercooling’ punte en tempo van waterverlies laer sal wees. Hierdie eienskappe kon egter nie by die individue geidentifiseer word nie. Ons het diapouse probeer induseer deur gebruik te maak van ‘n gesimuleerde oorgang na winterstoestande in die laboratorium. Die toestande het toegelaat vir korter dae en laer gemiddelde temperature gedurende beide die dag en nag. Die tweede doelwit waarna gekyk is, is die bepaling van VKM se beweging in die boorde en die vrugskade op verskillende vrugsoorte. Daar kon ‘n duidelike tendens geidentifiseer word in die toename van VKM hervangs by temperature bo ‘n minimum van 16°C en gemiddelde maksimum bo 30°C. Daar was 6 vrylatings periodes, met geen betekenisvolle getalle van hervangs nie. Daar was geen betekenisvolle verskille tussen die hervangsgetalle in die verskillende vrugsoorte nie, alhoewel die 30m lokval ‘n betekenisvol hoër gemiddelde hervangs gehad het, in vergelyking met lokvalle by 60m en 90m. Die hoeveelheid vrugskade is ook gemonitor op Granny smith appels, Forelle pere, Larry Ann pruime en Satsuma en Clementine sitrus kultivars. Die vrugte is na die laboratorium geneem waar die VKM tyd gegee is om uit te broei. Al die vrugsoorte het VKM volwassenes opgelewer, maar die eksperiment kon nie op appels in die laboratorium herhaal word tydens die toets van verkillende onwikkeling stadiums nie. Ons glo dus die VKM wat hier vanaf appels uitgebroei het, is weens sekondere infeksies in die boorde. Die hoogste skadetelling is in die sitrusboord gevind. Die derde doelwit was om die duur van onderskeie ontwikkeling stadiums te bepaal op vier vrugsoorte, sowel as op ‘n kunsmatige medium. Ons het ondervind dat die VKM nie Royal Gala of Pink lady kultivars kan infesteer onder laboratorium toestande nie. Die vrugsoorte wat dus ontwikkeling kon onderhou was Forelle pere, Clementines en Thompson pitlose druiwe. Die ontwikkeling vanaf eier na volwasse stadium was die kortste op druiwe, gevolg deur lemoene en pere. Die pere het die minste VKM onderhou in vergelyking met die lemoene en druiwe. Al die vrugte is binnegedring naby die aansluiting van die stingel aan die vrugte, behalwe die pere wat nader aan die kelk binnegedring is. Toekomstige navorsing sal gefokus moet word op die lewenssiklus in die veld, vir die verskillende vrugsoorte. Daar sal ook gekyk moet word na die spesifieke alternatiewe gashere of plekke waar die VKM kan oorwinter sodat beheer meer voorkomend plaas kan vind. Dit sal infestasie vlakke onderdruk, om veral laer druk tydens oesperiodes te verseker. Dit is uiters belangrik om beheer stategieë te kombineer met die hoeveelheid druk vanaf uitvoermarkte oor maksimum residu vlakke van chemiese middels.

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.