No description available.

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No description available.

The effect of protein in adult and larval diets for Ophyra aenescens (Wiedemann) (Diptera: Muscidae ), a facultative predator of the house fly, Musca domestica L., was examined. A larval diet of coarse wheat bran, vermiculite and dried meat powder was found adequate for rearing O. aenescens larvae. Protein content was important for maximum larval emergence and for F1 adult longevity and fecundity. Adult diet of powdered milk, sugar and dried meat powder was sufficient for maintenance of adult O. aenescens flies. Protein was needed in adult diet for optimal fecundity. Cyromazine was toxic to O. aenescens at high levels (1.0 ppm), but at 0.75 ppm O. aenescens tolerated cyromazine better than a susceptible strain of house flies. At these cyromazine levels, mass release of O. aenescens into poultry houses is possible one to two days after cyromazine has been removed from the chicken feed. The effect of temperature and manure moisture on O. aenescens was studied. At temperatures of 18°C, emergence of Ophyra aenescens was significantly lower than at temperatures of 21 and 27°C. Predation by O. aenescens at 27°C on house fly larvae was significant at ratios of 3:1, 2:1 and 1:1 (house fly to O. aenescens). A constant manure moisture was important in the development of O. aenescens larvae and its ability to prey on house fly larvae. At constant levels of 50, 60, and 70% manure moisture, O. aenescens substantially reduced house fly larval numbers at ratios of 3:1, 2:1 and 1:1. / Master of Science

Thesis (MScAgric (Conservation Ecology and Entomology)--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: The obscure mealybug, Pseudococcus viburni (Signoret) (Pseudococcidae), is one of the common and serious pests of apples and pears in South Africa. The management of this pest in South Africa is dominated by the use of insecticides, while research into using natural enemies for biological control of mealybugs is still ongoing. Increasing concern over the environmental impact, pesticide residues in fruits, resistance, and expense associated with frequent use of insecticides make it necessary to investigate alternative biological control methods, such as the use of entomopathogenic nematodes, for the control of mealybugs. Entomopathogenic nematodes have proven comparable or even superior to chemicals in controlling certain insect pests, without residue problems or a harmful effect on the environment. An important aspect of using endemic nematodes includes the identification of species of nematodes and their symbiotic bacterial cells. A study was carried out to describe a new species of Steinernema, which was recovered during a previous survey in citrus orchards in three provinces of South Africa. Morphometrics, morphology, crossbreeding, drawings, light microscopy and Scanning Electron Microscopy (SEM) photographs were used to describe the new species. A cryopreservation method has been simplified and optimised for the long-term storage of Steinernema khoisanae (SF87) and Heterorhabditis zealandica (J34). Different cryoprotectants used included 15% glycerol, 8% ethylene glycol and 8% dimethyl sulfoxide (DMSO), in which S. khoisanae was incubated at room temperature for periods of two, three, four and five days, followed by a methanol wash. An optimum survival rate of 69% was obtained for S. khoisanae after a four-day incubation period in 15% glycerol. This technique has been used for the cryopreservation of H. zealandica, with a 78% survival rate. The thawed nematodes of both species were able to infect Galleria mellonella larvae after 42 days of cryopreservation (-196ºC) and were able to complete their life cycles. / AFRIKAANSE OPSOMMING: Die ligrooswitluis, Pseudococcus viburni (Signoret) (Pseudococcidae), is een van die algemene en ernstige peste van appels en pere in Suid-Afrika. Die bestuur van hierdie pes word tans in Suid-Afrika deur die gebruik van insekdoders gedomineer terwyl navorsing oor die gebruik van natuurlike vyande vir die beheer van P. viburni nog aan die gang is. Die verhoogde kommer oor die omgewing, residue in vrugte, weerstand, en die koste verbonde aan die gereelde gebruik van chemiese middels maak dit nodig om alternatiewe biologiese metodes van beheer, soos die gebruik van entomopatogeniese nematodes vir die beheer van witluis, te ondersoek. In ander lande is reeds aangetoon dat entomopatogeniese nematodes onder sekere omstandighede en vir sekere insekte gelykwaardige of selfs beter beheer kan gee as chemiese middels. ʼn Belangrike aspek van die gebruik van endemiese nematodes vir die beheer van insekte sluit die korrekte identifikasie van die spesies met hul geassosieerde bakteriese simbionte in. ʼn Nuwe spesie van Steinernema is uit ʼn vorige opname van entomopatogeniese nematodes in sitrusboorde in drie provinsies van Suid-Afrika geïsoleer. Morfometrie, morfologie, kruisteling, ligmikroskoop en SEM fotografie is gebruik om ʼn nuwe spesies te beskryf. ʼn Kriopreserveringsmetode is ontwikkel en ge-optimaliseer vir die langtermyn bewaring van Steinernema khoisanae (SF87) en Heterorhabditis zealandica (J34). Verskillende kriobeskermingsmiddels insluitend 15% gliserol, 8% dimetiel sulfokied (DMSO) en 8% etileen glikol, waarin S. khoisanae vir periodes van twee, drie, vier, en vyf dae geïnkubeer is, is teen kamertemperatuur, getoets, gevolg deur ʼn metanolbad. Optimum oorlewing van 69% is verkry vir S. khoisanae nadat die infektiewe larwes (IJ) vir vier dae in 15% gliserol gehou is. Hierdie tegniek is ook toegepas op H. zealandica, met 78% oorlewing van die IJ. Die ontvriesde nematodes van beide spesies was in staat om Galleria mellonella larwes suksesvol te infekteer en hulle lewensiklus te voltooi nadat hulle vir 45 dae onder kriopreservering gehou is teen -196ºC.

False codling moth (FCM), Thaumatotibia leucotreta is an extremely important pest of citrus in South Africa and with the shift away from the use of chemicals, alternate control options are needed. One avenue of control which has only recently been investigated against the soil-borne life stages of FCM is the use of entomopathogenic fungi (EPF). In 2009, 12 entomopathogenic fungal isolates collected from South African citrus orchards showed good control potential during laboratory conducted bioassays. The aim of this study was to further analyse the potential of these isolates through concentration-dose and exposure-time response bioassays. After initial re-screening, concentration-dose response and exposure-time response sandconidial bioassays, three isolates were identified as exhibiting the greatest control potential against FCM in soil, Metarhizium anisopliae var. anisopliae (G 11 3 L6 and FCM Ar 23 B3) and Beauveria bassiana (G Ar 17 B3). Percentage mycosis was found to be directly related to fungal concentration as well as the amount of time FCM 5th instar larvae were exposed to the fungal conidia. LC50 values for the three isolates were not greater than 1.92 x 10⁶ conidia.ml⁻ₑ and at the LC₅₀, FCM 5th instar larvae would need to be exposed to the fungus for a maximum of 13 days to ensure a high mortality level. These isolates along with two commercially available EPF products were subjected to field persistence trials whereby net bags filled with a mixture of autoclaved sand and formulated fungal product were buried in an Eastern Cape citrus orchard. The viability of each isolate was measured on a monthly basis for a period of six months. All isolates were capable of persisting in the soil for six months with the collected isolates persisting far better than the commercially used isolates. Two of the isolates, G 11 3 L6 and G Ar 17 B3, were subjected to small scale laboratory application trials. Two formulations were investigated at two concentrations. For each isolate, each formulation and each concentration, FCM 5th instar larvae were applied and allowed to burrow into the soil to pupate before fungal application or after fungal application. Contact between fungi and FCM host is essential as, in contrast to pre-larval treatments, percentage mortality in post-larval treatments was low for both formulations and both isolates. For isolate G Ar 17 B3, a conidial suspension applied as a spray at a concentration of 1 x 10⁷ conidia.ml⁻ₑ obtained the highest percentage mortality (80 %). For isolate G 11 3 L6 however, both formulations performed equally well at a high, 1 x10⁷ conidia.ml⁻ₑ concentration (conidial suspension: 60 %; granular: 65 %) The results obtained thus far are promising for the control of FCM in citrus, but if these EPFs are to successfully integrate into current FCM control practices more research, some of which is discussed, is essential

The biology of key citrus pests Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae), Ceratitis capitata Wiedemann (Diptera: Tephritidae) and Ceratitis rosa Karsch (Diptera: Tephritidae) includes their dropping from host plants to pupate in the soil below citrus trees. Since most EP fungi are soil-borne microorganisms, the development and formulation of alternative control strategies using these fungi as subterranean control agents, targeted at larvae and pupae in the soil, can potentially benefit existing IPM management of citrus in South Africa. Thus, a survey of occurrence of entomopathogenic fungi was undertaken on soils from citrus orchards and natural vegetation (refugia) on conventionally and organically managed farms in the Eastern Cape Province in South Africa. A method for baiting soil samples with citrus pest T. leucotreta and C. capitata larvae, as well as with the standard bait insect, Galleria mellonella Linnaeus (Lepidoptera: Pyralidae), was implemented. Sixty-two potentially useful entomopathogenic fungal isolates belonging to four genera were collected from 288 soil samples, an occurrence frequency of 21.53%. The most frequently isolated entomopathogenic fungal species was Beauveria bassiana (Balsamo) Vuillemin (15.63%), followed by Metarhizium anisopliae var. anisopliae (Metschnikoff) Sorokin (3.82%). Galleria mellonella was the most effective insect used to isolate fungal species (χ2=40.13, df=2, P≤ 0.005), with a total of 45 isolates obtained, followed by C. capitata with 11 isolates, and T. leucotreta with six isolates recovered. There was a significantly (χ2=11.65, df=1, P≤ 0.005) higher occurrence of entomopathogenic fungi in soil samples taken from refugia compared to cultivated orchards of both organically and conventionally managed farms. No significant differences were observed in the recovery of fungal isolates when soil samples from both farming systems were compared. The physiological effects and host range of 21 indigenous fungal isolates obtained in the Eastern Cape were investigated in the laboratory to establish whether these isolates could be effectively used as biological control agents against the subterranean life stages of C. rosa, C. capitata and T. leucotreta. When these pests were treated with a fungal concentration of 1 x 10⁷ conidia ml⁻¹, the percentage of T. leucotreta adults which emerged in fungal treated sand ranged from 5 to 60% (F=33.295; df=21; P=0.0001) depending on fungal isolate and the percentage of pupae with visible signs of mycosis ranged from 21 to 93% (F= 96.436; df=21; P=0.0001). Based on fungal isolates, the percentage adult survival in C. rosa and C. capitata ranged from 30 to 90% and 55 to 86% respectively. The percentage of C. rosa and C. capitata puparia with visible signs of mycosis ranged from 1 to 14% and 1 to 11% respectively. Deferred mortality due to mycosis in C. rosa and C. capitata adult flies ranged from 1 to 58% and 1 to 33% respectively, depending on fungal isolate. Entomopathogenic fungal isolates had a significantly greater effect on the adults of C. rosa and C. capitata than they did on the puparia of these two fruit fly species. Further, C. rosa and C. capitata did not differ significantly in their response to entomopathogenic fungi when adult survival or adult and pupal mycosis were considered. The relative potency of the four most virulent Beauveria isolates as well as the commercially available Beauveria bassiana product, Bb Plus® (Biological Control Products, South Africa), were compared against one another as log-probit regressions of mortality against C. rosa, C. capitata and T. leucotreta which all exhibited a dose-dependent response. Against fruit flies the estimated LC50 values of all five Beauveria isolates ranged from 5.5 x 10¹¹ to 2.8 x 10¹² conidia/ml⁻¹. There were no significant differences between the relative potencies of these five fungal isolates. When T. leucotreta was considered, isolates: G Moss R10 and G 14 2 B5 and Bb Plus® were significantly more pathogenic than G B Ar 23 B3 and FCM 10 13 L1. The estimated LC₅₀ values of the three most pathogenic isolates ranged from 6.8 x 10⁵ to 2.1 x 10⁶ conidia/ml⁻¹, while those of the least pathogenic ranged from 1.6 x 10⁷ to 3.7 x 10⁷ conidia/ml⁻¹. Thaumatotibia leucotreta final instar larvae were exposed to two conidial concentrations, at four different exposure times (12, 48, 72 and 96 hrs) and showed an exposure time-dependant relationship (F=5.43; df=3; P=0.001). At 1 x 10⁷conidia/ml⁻¹ two Beauveria isolates: G Moss R10 and G 14 2 B5 were able to elicit a response in 50% of test insects at 72 hrs (3 days) exposure. Although a limited amount of mycosis was observed in the puparia of both fruit fly species, deferred adult mortality due to mycosis was high. The increased incidence of adult mortality suggests that post emergence mycosis in adult fruit flies may play a more significant role in field suppression than the control of fruit flies at the pupal stage. The increased incidence of pupal mortality, as well as the relatively low concentrations of conidia required to elicit meaningful responses in T. leucotreta pupae may suggest that pre-emergent control of false codling moth will play a more significant role in field suppression than the control of adult life stages using indigenous isolates of entomopathogenic fungi. Various entomopathogenic fungal application techniques targeted at key insect pests within integrated pest management (IPM) systems of citrus are discussed.

The diamondback moth, Plutella xylostella (Linnaeus 1758.) (Lepidoptera: Plutellidae), is a major insect pest of Brassica crops in many parts of the world leading to economic losses amounting to an estimated US$ 4-5 billion. Although parasitoids (biological control agents) play a major role in suppressing the pest populations during November – May in South Africa, the pest reaches outbreak status during September and October due to low impact of parasitoids, which has necessitated regular application of insecticides. However, insecticide applications have often resulted in the pest developing resistance, and when coupled with the negative effects of several insecticides on parasitoids, integration of the two pest control strategies for effective management of P. xylostella population density has been difficult to achieve. One approach that has received little attention is integration of host plant resistance (bottom-up effect) and biological control (top-down effect) for effective management of P. xylostella. However, the interaction between host plants, the insect pest, and parasitoids is not simple and straight forward, as strong negative impact of host plants on fitness of the insect pest can be cascaded up the food chain and have a negative impact on a given parasitoid, which in turn may reduce the desired complementary effect between the two pest control strategies. To identify optimal interactions between cabbage (Brassica oleracea L. var. capitata, Brassicaceae), P. xylostella and its larval parasitoid Cotesia vestalis (Haliday 1834) (Hymenoptera: Braconidae), this study investigated (i) the effects of seven cabbage cultivars (Empowa, Hollywood F1, Megaton, Leano, Menzania, Beverley Hills and Karabo) on fitness parameters (survival, developmental time, pupal weights, longevity without food and oviposition rates) of P. xylostella; (ii) the influence of the same host plant cultivars on fitness parameters (developmental time, pupal weights, longevity xi without food, fecundity, emergence rate and sex ratio) of C. vestalis. Furthermore, net reproductive rates and the intrinsic rates of natural increase were calculated for C. vestalis that emerged from hosts fed on each of the cultivars. All experiments were conducted in climate-controlled laboratory rooms maintained at 22 ± 1 ºC (mean ± S.D.), 60 ± 5 % RH and 16L: 8D photoperiod. Under the no choice test, overall survival of P. xylostella immature stages was highest on Karabo (67.26%) and lowest on Megaton (44.92%). The larval and pupal developmental period, and generation time was prolonged on Empowa (18.48 days), Karabo (14.64 days) and Beverly Hills (17.48 days), while developmental period on Hollywood F1 (13.79 days) was shortest. Male and female P. xylostella pupal weights were lighter from larvae that fed on Megaton (4.13 and 4.65 mg), Menzania (4.53 and 4.91 mg), and Hollywood F1 (4.11 and 5.08 mg), whereas pupal weights from Karabo (6.0 and 6.82 mg) were the heaviest. Unfed female moths originally reared on Beverley Hills had the highest longevity (5.05 days), whereas those reared on Leano (3.54 days) and Megaton (3.89 days) had the shortest life span. Under the choice-test, P. xylostella moth laid significantly more eggs on Empowa (48.8%) and Hollywood F1 (45.6%) and least on Menzania (11.8%) and Leano (10.6%). Megaton was more resistant to P. xylostella due to lower survival rates of immature stages, lower pupal weights and moth longevity. The generation time of C. vestalis was shortest on Karabo (10.10 days) and Leano (10.38 days), and longest on Megaton (12.57 days) and Empowa (12.80 days). The highest pupal weight of C. vestalis was obtained from parasitoids reared from P. xylostella fed Menzania (5.4 mg), Megaton (5.25 mg) and Beverly Hills (4.85 mg) and the lightest on Karabo (3.8 mg). Parasitoids reared on larvae that fed on Hollywood F1 lived the longest (2.28 days) followed by Menzania (1.94 days) and Beverly Hills (1.8 days), whereas those whose hosts fed on Leano had shortest life span (0.83 days). Despite the parasitoids from Megaton hosts being heavier, their fecundity and number of female progeny per female (16.87 and 3.60, respectively) were lowest. Cotesia vestalis fecundity and daughters produced per female were highest on hosts fed on Menzania (38.00 and 9.13, respectively) and Beverly Hills (32.87 and 9.07, respectively). As a consequence, the net reproductive rate (R0) and intrinsic rate of increase (r) were higher on Menzania (7.87 and 0.58, respectively) and Beverly Hills (8.29 and 0.62, respectively). As survival and overall fitness of P. xylostella was lower on Megaton, this cultivar can play a major role in restricting population growth of this pest and thus generational number of eggs deposited on it during September and October. However, this strong bottom-up effect of Megaton on P. xylostella was cascaded up the food chain, as overall fitness of C. vestalis was lower on hosts developing on it. In contrast, the overall fitness of C. vestalis was higher on hosts that developed on Menzania and Beverly Hills. As these cultivars showed potential to sustain population density of C. vestalis at higher levels, it is also assumed that the period required for the parasitoid to reach the critical density to suppress the host population at a lower average density will be reached quicker than on other cultivars. Thus, their cultivation may improve biological control of P. xylostella during November–May in South Africa / Agriculture, Animal Health and Human Ecology / M. Sc. (Agriculture)

Thesis (MScAgric)--Stellenbosch University, 2012. / 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. / AFRIKAANSE OPSOMMING: Die landboubedryf in Suid-Afrika word oorheers deur die gebruik van insekdoders. Vervaardigers steun swaar op chemikalieë wat toenemend gesondheids-, omgewings- en ekologiese risiko's, asook die snelle ontwikkeling van weerstand in sleutelinsekteplae veroorsaak, en wat reste van plaagdoders op gewasse laat. Na aanleiding van toenemende besorgdheid oor die impak van hierdie plaagbestuurspraktyke op die omgewing, word alternatiewe plaagbestuurstrategieë ondersoek. Entomopatogeniese nematodes (EPNs) is geïdentifiseer as belowende biologiese beheeragente van sleutelinsekteplae. Die twee EPN genera wat belofte inhou vir gebruik as biologiese beheeragente binne 'n geïntegreerde plaagbestuursprogram is Steinernema en Heterorhabditis. Kommersialisering en die geslaagde gebruik van EPNs om insekplae te beheer in Noord-Amerika, Australië, Europa en Asië, het die doeltreffendheid van hierdie organismes as biologiese beheeragente bevestig. Ongelukkig is EPNs in groot genoeg getalle vir kommersiële aanwending in die veld nog nie op die Suid-Afrikaanse mark beskikbaar nie. Groot getalle EPNs kan deur in vivo en in vitro teling verkry word. Die doelwit van hierdie studie was om die in vivo produksieproses te stroomlyn deur die gebruik van twee endemiese EPN spesies, Heterorhabditis zealandica (SF41) en H. bacteriophora (SF351). Hierdie EPN isolate is deur navorsing bewys om doeltreffende beheeragente van kodlingmot Cydia pomonella, vals kodlingmot Thaumatotibia leucotreta, ligrooswitluis Pseudococcus viburni, en gebande vrugtekalanders Phlyctinus callosus te wees. 'n Vergelykende studie is gedoen om geskikte gasheerinsekte vir EPN produksie van plaaslike H. zealandica (SF41) en H. bacteriophora (SF351) isolate te vind. Gashere is geselekteer op grond van vatbaarheid vir die EPN spesie wat gebruik word, en algemene beskikbaarheid en gemak en koste van teling. Wasmotlarwes Galleria mellonella (WML) en meelwurms Tenebrio molitor (MW) is as gashere gekies. Ten einde nematodes van konsekwente kwaliteit te teel, word 'n deurlopende bron van gasheerinsekte benodig wat op 'n gestandaardiseerde dieet voed. WML en MW is onderskeidelik op vyf verskillende diëte geteel by ±26°C in die donker. Die beste dieet vir elke gasheer is gekies op grond van die dieet wat, gemiddeld, die swaarste larwes binne 'n spesifieke tydsraamwerk opgelewer het. Die swaarste WML, teen 'n gemiddelde massa van 0.19 g per larwe, is geteel op 'n dieet wat bestaan het uit 118 g koringmeel, 206 g semels, 118 g melkpoeier, 88 g brouersgis, 24 g verpoeierde was, 175 ml heuning en 175 ml gliserol. Die swaarste MW larwes het gemiddeld 0.0154 g per larwe geweeg en is geteel op 'n dieet van 100% semels. Ten einde die hipotese te bevestig dat 'n lineêre verwantskap bestaan tussen die massa van 'n insekgasheer en die aantal nematodes wat deur daardie gasheer geproduseer word, is 'n studie gedoen om die aantal H. zealandica en H. bacteriophora per gasheergram te bepaal. WML, MW, kodlingmotlarwes en vals kodlingmotlarwes is individueel geweeg en met infektiewe larwes van die twee onderskeidelike EPN spesies geïnokuleer. Daarbenewens is die vermeerdering van nematodes in bevrore MW en WML ook getoets. Die aantal nematodes wat in elke gasheer geoes is, is getel, en die gemiddelde nematode-afstammelinge in elke gasheer bereken. 'n Beduidende lineêre korrelasie tussen die massa van WML en MW en die aantal H. zealandica en H. bacteriophora wat onderskeidelik geproduseer is, het die hipotese bevestig dat nematode-vermeerdering binne hierdie gashere toeneem namate die gasheermassa toeneem. WML het die meeste H. zealandica en H. bacteriophera per gasheergram opgelewer (1 459 205± 113 670 en 1 898 512± 94 355 onderskeidelik), gevolg deur MW larwes (836 690± 121 252 en 414 566± 67 017 onderskeidelik). Laer getalle H. zealandica and H. bacteriophora per gram kodlingmot (57 582 ± 10 026 en 39 653 ± 8 276) en per gram vals kodlingmot (192 867 ± 13 488 en 97 652 ± 23 404) is egter geproduseer. Een van die sleutelfaktore vir die doeltreffendheid van die in vivo vermeerdering van nematodes is geslaagde gasheerinfeksie. Drie inokulasietegnieke is dus geëvalueer en vergelyk deur H. zealandica en H. bacteriophora te gebruik: inokulasie met 'n pipet, skud van gashere in 'n nematode-inokulum, en gasheerindompeling in 'n nematode-suspensie. WML en MW is as gashere gebruik vir elke inokulasietegniek, en is geïnokuleer met nematodes wat uit 'n konsentrasie van 200 infektiewe larwes (ILs) / insek larwe bestaan het. Die persentasie dooie insekgashere is na twee dae bepaal, en infeksie soos bevestig deur kleurverandering en disseksie, na sewe dae. Die hoogste persentasie infeksie deur sowel nematode-isolate as gashere te gebruik, was met die pipet-tegniek. Die infeksiekoerse vir alle nematode-gasheerkombinasies met die pipet-tegniek was hoër as 90%, met die uitsondering van MW-inokulasie met H. bacteriophora, waar die infeksie 76% was. Hierdie studie toon dat afwykings voorkom in die mate van gasheerinfeksie, na gelang van die inokulasietegniek wat gebruik is. In 'n bykomende poging om infeksie na inokulasie te verhoog, is H. zealandica, H. bacteriophora en MW onderwerp aan stressors en bymiddels om nematode-infeksie te bevorder. Drie behandelings, asook 'n kontrole-behandeling, is vergelyk. Infeksievlakke het nie verhoog deur MW voor inokulasie aan kraanwater van 70°C bloot te stel nie. Inteendeel, laer infeksievlakke is opgemerk waar gashere in kraanwater van 70°C gedompel is en daarna met H. bacteriophora geïnokuleer is, vergelyke met die kontrole. Gasheerinfeksie van slegs 12% is verkry, vergelyke met 48% in die kontrole. Infeksie van 21% is met H. zealandica verkry. Die virulensie van nematodes het nie beduidend toegeneem deur H. zealandica en H. bacteriophora IL in 'n suspensie met Mn2+SO4H20 te behandel voor MW geïnokuleer is nie. Inokulasie van MW met behandelde H. zealandica IL het tot 'n infeksie van 81% gelei, vergelyke met die kontrole waar 'n infeksie van 80% behaal is. H. bacteriophora het 'n MW-infeksie van 47% veroorsaak, vergelyke met die kontrole se infeksiekoers van 48%. 'n Kombinasie van die twee bogenoemde behandelings het eweneens nie gasheerinfeksievlakke verhoog nie. Die indompeling van meelwurms in kraanwater van 70°C voor inokulasie met nematodes wat met Mn2+SO4H20 behandel is, het tot gasheerinfeksie van 13% en 9% onderskeidelik gelei wanneer H. bacteriophora en H. zealandica gebruik is. Toekomstige navorsing is nodig om die protokol te verbeter wat in hierdie studie gebruik is om MW en plaaslike nematode-isolate aan stressors te onderwerp. 'n Ondersoek is gedoen na die vermoë van twee formulasies om biologiese aktiwiteit en virulensie van H. zealandica te onderhou. 'n Kwaliteitsstandaardtegniekis gebruik om weekliks die persentasie oorlewing en virulensie van geformuleerde H. zealandica oor 'n tydperk van 21 dae te meet. IL is in Pesta korrels en klappervesel geformuleer, terwyl nematodes in kraanwater gedien het as kontrole. Die aantal lewende H. zealandica in Pesta korrels en klappervesel het drasties verminder na sewe dae in die formulasie. Oorlewing van nematodes in Pesta korrels het gedaal tot 9.79% na 21 dae vergyleke met die kontrole, waar 79.79% oorleef het. Nog minder nematodes - 25.84% - het na sewe dae in die klappervesel oorleef, en slegs 2.25% na 21 dae. Na 21 dae van berging het 100%+ van nematodes oorleef in die kontrole vir klappervesel. Die toepassing van die kwaliteitsstandaardtegniek om die virulensie van geformuleerde H. zealandica te bepaal, het ondoeltreffend geblyk. Verhoogde MW sterftesyfers is verkry in die kontrole waar geen nematodes by die inseklarwes gevoeg is nie, vergelyke met die byvoeging van hoër dosisse nematodes. Nietemin, die aanpassing van sekere aspekte in die protokol van hierdie kwaliteitsbeheermeting om spesifiek plaaslike toestande in ag te neem, sou dit moontlik 'n meer doeltreffende middel kon maak om die virulensie van endemiese nematodes te bepaal.

Thesis (MSc)--Stellenbosch University, 2003. / Full text to be digitised and attached to bibliographic record. / ENGLISH ABSTRACT: The losses incurred to by the South African hardwood industry because of damage caused by the larvae of the Australian eucalyptus borers Phoracantha semipunctata (Fabricius) and P. recurva Newman (Coleoptera: Cerambycidae) were countered by the introduction of various biological control agents. Megalyra fasciipennis Westwood (Hymenoptera: Ichneumonidae), restricted to the southwestern Cape for nearly 91 years after its probable establishment in 1910, is a specialist pupal parasitoid achieving a parasitism level of up to 52.5 %. It has an activity peak in early spring, which coincides with the pupation of a large percentage of its hosts that had overwintered as larvae. The average length of the ovipositor of M. fasciipennis (42.71 ± 5.33 mm S.D.) was longer than the average tunnel length (31.34 ± 11.85 mm S.D.) to the pupal chamber of Phoracantha spp, within the log despite variations in the thicknesses of the eucalypt stems. Stem thickness therefore did not adversely affect the level of parasitism. Megalyra fasciipennis adults are diurnal, with activity largely determined by temperature. Over 70 % were active between 25°C and 34 °C, the minimum threshold for activity being 16°C. Optimum temperature for oviposition was 30 °C. Males became active before the females and maximum oviposition occurred between 10hOOand 15hOO. In 1993, a host specific egg parasitoid, Avetianella longoi Siscaro (Hymenoptera: Encyrtidae), was introduced for the control of Phoracantha spp. A total of 7791 A. longoi adults and 80 parasitised eggs were released around Cape Town between 1993 and 1995 before establishment was confirmed. Dispersal was monitored annually and was determined to occur at a rate of 50 km/year. By 1998 A. longoi had dispersed 300 km north of Cape Town to Lutzville and 270 km east to Riversdal. Subsequent to a satellite release in Knysna during 1994, it has been established 40 km from this release site, at Plettenberg Bay. The parasitoid has also bridged a LO km expanse of ocean to establish on Robben Island, immediately off the west coast of Cape Town. Avetianella longoi has a preference for the eggs of P. semipunctata, which is the most likely cause for the decline in the population of P. semipunctata. However, P. recurva remains relatively unaffected. Average parasitism of Phoracantha spp. eggs by A. longoi was 59.4 %. An undescribed Cleonymus sp. (Hymenoptera: Pteromalidae) of unknown origin (the genus being widely distributed on several continents), was discovered in the Cape Peninsula parasitising late instar larvae of P. semipunctata and P. recurva. This ectoparasitoid lays its eggs (mean number per larva = 20.3 ± 15.2 S.D.) through the bark into the host chamber after the host has been paralysed. The host is entirely consumed and pupation takes place in the chamber with wasps emerging in the ratio of 1 male: 3 females. Although uncommon in the field, mass rearing of these wasps in culture was easily accomplished and a consignment was released in the Tzaneen district in 1993, where it was confirmed to have become established in 1996. Bark thickness constraints on the effectiveness of this parasitoid as a biological control agent because it's short ovipositor restricts the wasp to certain eucalypt species or trees with thin bark. The introduction into South Africa in 1995 and attempted establishment of the larval parasitoids, Syngaster lepidus Brullé (Hymenoptera: Braconidae), Jarra phoracantha Marsh & Austin (Hymenoptera: Braconidae) and J maculipennis Marsh & Austin proved unsuccessful in the Western Cape. However, the former two species were established in the Tzaneen district and their recruitment for release in the Western Cape should be considered. The present guild of biological control agents has been insufficient to give the required control. In the absence of biological control agents, intraspecific competition amongst host larvae is the major mortality factor. Although high levels of mortality are achieved as a result of parasitism despite the biological constraints of the parasitoids (e.g. the narrow activity peak of A. longoi and the restriction of Cleonymus sp. to thin barked eucalypts), their combined parasitism has succeeded in reducing the competition between host larvae, resulting in fewer yet larger host beetles emerging. The introduction of additional viable agents to assist in the biocontrol of Phoracantha spp. is required. / AFRIKAANSE OPSOMMING: Die verliese wat die Suid-Afrikaanse hardehoutbedryf ly as gevolg van skade veroorsaak deur die bloekomboorders Phoracantha semipunctata en P. recurva (Coleoptera: Cerambycidae), is bekamp deur die invoer van verskeie biologiese beheeragente. Megalyra fasciipennis Westwood (Hymenoptera: Ichneumonidae), beperk tot die Suidwes-Kaap vir byna 91 jaar nadat dit waarskynlik in 1910 daar gevestig is, is In spesialis papieparasitoïd wat In parasitismevlak van tot 52.5% bereik. Dit het In aktiwiteitspiek in die vroeë lente wat saamval met die papievorming van baie gasheerlarwes wat oorwinter het. Die gemiddelde lengte van die eierboor van M. fasciipennis (42.71 ± 5.33 mm S.A.) was langer as die gemiddelde tonnellengte (31.34 ± 11.85 mm S.A.) na die papieholte van die gasheer binne in die hout, ten spyte van die variasie in die dikte van die bloekomstamme. Stamdikte het dus nie In nadelige uitwerking op die vlak van parasitisme nie. Volwassenes van M.fasciipennis is bedags aktief en aktiwiteit word hoofsaaklik deur temperatuur bepaal. Meer as 70% was tussen 25°C en 34 °C aktief, met 16°C as die minimum drumpel vir aktiwiteit. Mannetjies het voor die wyfies aktief geword en maksimum eierlegging het tussen 10hOOen 15hOO plaasgevind. In 1993 is die gasheerspesifieke eierparasitoïd Avetianella longoi Siscaro (Hymenoptera: Encyrtidae) vir die beheer van Phoracantha spp. ingevoer. Van 1993 tot 1995 is 7791 volwassenes van A. longoi en 80 geparasiteerde eiers rondom Kaapstad vrygelaat en dis vasgestel dat die spesies gevestig het. Die verspreiding daarvan is jaarliks gemonitor en dis vasgestel dat dit teen 50 km per jaar plaasvind. Teen 1998 het dit versprei tot 300km noord van Kaapstad na Lutzville en 270 km oos na Riversdal. Na 'n satelliet-loslating by Knysna in 1994 het dit 40 km verder by Plettenbergbaai gevestig. Die parasitoïd het ook 10 km van die oseaan oorgesteek om op Robbeneiland, wes van Kaapstad te vestig. Avetianella longoi gee voorkeur aan die eiers van P. semipunctata en dis waarskynlik die rede vir die afname in die getalle van hierdie spesies, maar P.recurva word relatief min beïnvloed. Die gemiddelde graad van parasitisme van Phoracantha spp. was 59.4%. Dit is gevind dat 'n onbeskryfde Cleonymus sp. (Hymenoptera: Pteromalidae) van onbekende oorsprong (die genus kom wyd verspreid in verskeie vastelande voor) die laat instar larwes van P. semipunctata en P. recurva parasiteer. Hierdie ektoparasitoïd lê sy eiers (gemiddeld 20.3 ± 15.2 S.A.) in die gasheerholte nadat die gasheer eers verlam is. Die gasheer word heeltemalopgevreet en pupering vind plaas in die holte plaas. Volwassenes kom uit in verhoudingvan drie mannetjies tot een wyfie. Alhoewel skaars in die natuur, kan hierdie wesp maklik in massa geteel word. 'n Besending is in die Tzaneen distrik vrygestel en in 1996 is vasgestel dat hulle gevestig het. Basdikte is 'n beperkende faktor in die gebruik van hierdie parasitoïd as effektiewe beheeragent vir biologiese beheer omdat die kort lengte van die eierboor die wesp sal beperk tot bloekomsoorte met dun bas. Die invoer na Suid-Afrika in 1995 en vestiging van die larwale parasitoïde Syngaster lepidus Brullé (Hymenoptera: Braconidae), J. phoracantha Marsh & Austin (Hymenoptera: Braconidae) en J. maculipennis Marsh & Austin was onsuksesvol in dieWes-Kaap. Die twee spesies is egter in die distrik Tzaneen gevestig en versameling met die oog op loslating in die Wes-Kaap behoort oorweeg te word. Die huidige gilde van biologiese beheer-agente is onvoldoende om die vereiste graad van beheer te verskaf. In die afwesigheid van biologiese beheer-agente is intraspesifieke kompetisie tussen gasheerlarwes die belangrikste mortaliteitfaktor. Alhoewel hoë vlakke van mortaliteit as gevolg van parasitisme bereik word, ten spyte van die biologiese beperkings van die parasitoïde (bv. die kort aktiwiteitspiek van A. longoi en die beperking van Cleonymus tot bloekoms met dun bas), het die gekombineerde parasitisme daarin geslaag om die kompetisie tussen gasheerlarwes te verlaag, met die gevolg dat minder maar groter gasheerkewers verskyn het. Dit is dus nodig dat addisionele organismes gevestig word om by te dra tot die biologiese beheer van Phoracantha spp.