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

There is a growing recognition globally that many agrochemicals are hazardous to humans, animals and the environment. Therefore, there is a need to substitute these chemical products with biological and physical treatments, and to change agronomic practices in order to control pests and diseases in agriculture. The primary objective of this thesis was to develop and test in laboratory, field and commercial packhouses trials as alternative control measures against green mould of citrus (caused by Penicillium digitatum Pers: Fr. Sacc) and Penicillium molds of litchi (caused by several Penicillium). A South African isolate of P. digitatum, isolated from an infected orange fruit, was found to be resistant to imazalil (the standard postharvest fungicide used in South Africa). Sixty yeast and 92 Bacillus strains were screened for their antagonistic activity against this isolate of P. digitatum. None of the yeasts or Bacillus isolates produced a curative action against P. digitatum on oranges. However, yeast Isolate B13 provided excellent preventative control of P. digitatum, superior to all the Bacillus isolates, when it was applied to citrus fruit prior to artificial inoculation with P. digitatum. Electron microscopy showed that yeast Isolate B13 inhibited conidial germination of P. digitatum. For the control of P. digitatum pre-harvest, trees were sprayed with a yeast, Isolate B13, a few months or a few days before harvest. However, this treatment alone proved to be ineffective in providing preventative control of green mould on Valencia oranges. For the control of P. digitatum preharvest, trees were treated with potassium silicate for a full season. Regular potassium silicate treatments resulted in a significant preventative control of P. digitatum infection on both navel and Valencia oranges. Treatment of Eureka lemons with potassium silicate as a postharvest treatment for the control of P. digitatum resulted in reduced disease lesion diameters when applied preventatively or curatively. Electron microscopy showed that potassium silicate inhibited germination of P. digitatum conidia and growth of its mycelium. Hot-water dip treatment at 50-58°C for 60-180 seconds (in increments of 15 seconds), significantly reduced infection development in inoculated wounds of Valencia oranges compared with control fruit treated with tap water, without causing any rind damage. The integration of the yeast, a hot water dip and potassium silicate pre-and postharvest applications provided control of P. digitatum control in multiple packhouse trials. The control achieved by the yeast Isolate B13 or hot-water, and potassium silicate in the packhouse alone was superior or equivalent to that provided by imazalil. A similar study was also carried out to determine possible control measures for Penicillium sp. on litchis. In this study, a total of 23 yeast and 13 Bacillus isolates were obtained from litchi fruit surfaces. Ten yeast and 10 Bacillus isolates that had shown good efficacy against P. digitatum of citrus were added to these for screening against Penicillium sp. of litchis. None of the yeasts or Bacillus isolates produced a curative action against Penicillium sp. infection on litchis. However, several yeast isolates (YL4, YL10, YLH and B13) resulted in reduced severity of the pathogen, when applied preventatively, compared with an untreated control. The yeast isolates were superior to all the Bacillus isolates, when applied to litchis prior to artificial inoculation by Penicillium infection on litchis. Potassium silicate as a postharvest treatment for the control of the pathogen caused reduced lesion diameters when applied preventatively or curatively to naturally infected litchis. The results presented in this thesis highlight the use of biological, physical and agronomic practices singly or in combination as an alternative control strategy against citrus postharvest green mould. This thesis also provides an insight into expanding these strategies, partly or fully, for the control of other postharvest Penicillium infections using litchi as an example. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

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.

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.

Includes bibliographical references (leaves 102-114) The highly successful biological control of the citrophilous mealybug Pseudococcus calceolarie (Maskell) (CM) by the parasitic wasp Coccophagus gurneyi Compere in several countries led to the release of this parasitoid in the Riverland of South Australia as part of an integrated pest management program. However CM has not been successfully controlled in this region. The results of this study may help to explain the lack of effective biological control of CM in Riverland citrus.

Lantana camara L. (Verbenaceae) remains a highly invasive and ecologically damaging weed in South Africa, despite some 50 years of biological control efforts. Lack of success has been ascribed to varietal differences, climate and predation of agents but these have not been tested. In this study, the effects of ant predation were tested on populations of three biological control agents for L. camara. Colonies of two species, Crematogaster sp. 1 and 2 were investigated. Crematogaster sp. 1 colonies were offered no choice between immature stages of the agents Hypena laceratalis Walker (Lepidoptera: Noctuidae), Falconia intermedia Distant (Hemiptera: Miridae) or Teleonemia scrupulosa Stål (Hemiptera: Tingidae) on lantana shoots. Density-dependent predation on F. intermedia and T. scrupulosa nymphs on lantana shoots was tested using Crematogaster sp. 2 colonies. In choice experiments Crematogaster sp. 2 colonies were offered F. intermedia or T. scrupulosa nymphs on potted lantana plants. Preliminary food trials confirmed that colonies foraged for protein, thereby validating results of no-choice experiments. Crematogaster sp.1 foragers removed 50% of F. intermedia nymphs, followed by 45% of H. laceratalis larvae and only 9% of T. scrupulosa nymphs. Foragers recruited most actively to H. laceratalis larvae and significantly more H. laceratalis biomass was removed than either F. intermedia or T. scrupulosa. A trade-off existed in prey size selection because larger larvae provided considerably more biomass but required forager cooperation and a longer time to subdue than did smaller prey. This increases both forager energy expense and mortality risk by other predators. This study showed that all Crematogaster sp. 1 colonies removed small (≤10mm) H. laceratalis larvae more frequently than larvae larger than 10mm. Thus, of these biological control agents, predators probably prefer small H. laceratalis larvae. Significantly more F. intermedia than T. scrupulosa nymphs were removed by Crematogaster sp. 1, while Crematogaster sp. 2 colonies removed comparable numbers of both agent species. Falconia intermedia nymphs' fast movement triggered a predatory response by these ant species. In contrast, the relatively immobile behaviour of T. scrupulosa nymphs was identified as a highly effective predator avoidance strategy. Since T. scrupulosa nymphs are unable to escape predators by moving, they appear to depend on the presence of alternative prey attracting predator attention. At high agent and/or forager density, T. scrupulosa nymphs attempted escape, but foragers identified them as prey once they moved and caught them. Predation on F. intermedia was also density dependent in that at high nymph and/or forager densities, escape routes were congested and nymphs were more easily caught. Survival of F. intermedia and T. scrupulosa nymphs in particular was low on ant-accessed shrubs in choice experiments and high on ant-excluded shrubs. It is likely that ants significantly depress F. intermedia populations in the field since besides predation, ant foragers probably interrupt F. intermedia feeding and ovipositioning. The combination of parasitism and predation on early instar larvae may explain why H. laceratalis occurs across lantana's range in South Africa but populations remain low. It is unlikely that T. scrupulosa nymphs are habitually preyed on by ant species unless they attract attention by being mobile. Although biological control of L. camara is influenced by climate and physiological defence mechanisms, this study has shown that predation by two ant species severely impacts leaf-feeding agents for L. camara. Thus, it is recommended that future selection of additional agents to control lantana should exclude leaf-feeding .

Pereskia aculeata Miller (Cactaceae) is an environmental weed that is damaging to natural ecosystems in South Africa. The plant is native to Central and South America and was first recorded in South Africa in a botanical garden in 1858. In this thesis, research into the biological control of P. aculeata was conducted with the intention of improving the control of the weed. A pre-release study of the relationship between P. aculeata density and native plant biodiversity indicated that P. aculeata has a negative impact on native biodiversity. The native plant biodiversity associated with different P. aculeata densities was used to determine threshold values and goals for the control of the weed. A threshold value of 50% P. aculeata density was calculated, indicating that P. aculeata density must be maintained below 50% in order to conserve native plant biodiversity. The ultimate goal of the control programme should be to maintain P. aculeata densities below 30%. At these densities there was no significant difference in native plant biodiversity from if the weed were absent from the ecosystem. The biological control agent, Phenrica guérini Bechyne (Chrysomelidae), has been released in South Africa but the potential of the agent to impact P. aculeata is not known and no post release evaluation has been conducted. Impact assessment studies indicate that P. guérini does not impact P. aculeata, even at high densities, but the results of greenhouse experiments should be interpreted with caution because of problems with extrapolation into the field. Although observations in the field suggest that P. guérini has reduced P. aculeata densities at one site, it is clear that new biological control agents are needed to reduce the weed to acceptable levels. Identifying the origin of the South African P. aculeata population was believed to be important to the biological control programme due to the disjunct native distribution and intraspecific variation of the species. Natural enemies associated with plant genotypes in different parts of the native distribution may have developed specialised relationships with certain intraspecific variants of the plant, resulting in differences in agent efficacy on certain host plant genotypes. A molecular study indicated that the closest relatives to the South African weed population found in the native distribution were in Rio de Janeiro Province, Brazil. A bioassay experiment in which fitness related traits of the biological control agent, P. guérini, were measured on various P. aculeata genotypes was conducted to determine the importance of host plant intraspecific variation. There was little variation in fitness traits between genotypes and no evidence of intraspecific host plant specialization. Although intraspecific variation had no effect on agent efficacy in the case of P. guérini, it is possible that other natural enemies may be more specialized. Genotype matching is expected to be more important when natural enemies likely to be specialised to individual genotypes are considered for biological control. Potential biological control agents were prioritized from data collected on surveys in the native distribution. The most promising of these, based on the presence of feeding, incidence, predicted host range, climatic matching, genotype matching and mode of damage, are two species of Curculionidae, the current biological control agent P. guérini and the stem boring moth, Maracayia chiorisalis Walker (Crambidae). The two curculionid species and M. chlorisalis should be considered priorities for host specificity studies. Releases of P. guérini and any new biological control agents should be made at sites where the pre-release study was conducted so that post-release evaluation data can be compared with the pre-release data and the impact of biological control can be evaluated. Retrospective analyses of biological control programmes provide important ways of improving aspects of biological control programmes, such as methods of agent selection. The evaluation of success in biological control programmes is essential for retrospective analyses because factors that have lead to successes or failures can be analysed. Retrospective analyses of biological control programmes, such as this thesis, may improve weed management, thereby contributing to the conservation of natural resources.

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

Potential vegetative and reproductive increases in yield, as well as the biological efficacy against Fusarium verticillioides and F. proliferatum causing ear and stem rot in maize crops of commercially-formulated micro-organism formulation T-Gro (Trichoderma harzianum isolate DB103 WP) combined with Spartacus (Beauveria bassiana isolate DB 105 WP), T-Gro combined with Armenius (Bacillus subtilis isolate DB 109 WP), T-Gro combined with Maximus (Bacillus subtilis isolate DB 108 WP), T-Gro combined with Shelter (Bacillus subtilis isolate DB 101), T-Gro combined with Bismarck (Microbacterium maritypicum isolate DB 107 WP), as well as individual treatments of T-Gro, Armenius, Bismarck, Maximus and Shelter, were investigated when applied to maize seed and soil under field conditions. All the micro-organism treatments were compared with Thiram 750WP (750g/kg thiram WP) and an untreated control. The micro-organism treatments showed an increase in vegetative as well as reproductive yields when compared to the reference product Thiram 750 WP and the untreated control. There were no observations of adverse effects on the germination of maize seed in all the treatments that were applied. The three isolates B. subtilis, T. harzianum, and M. maritypicum, showed a significant reduction in vascular tissue discolouration of the main and ear stems, indicating a potential to be used in the reduction and control of diseases caused by Fusarium spp. Results also showed poor to very good increases of stem and foliage biomass as well as cob yield per plant produced by the micro-organism treatments when compared to the untreated control. The highest cob yield per plant that differed significantly from the untreated control was produced by T-Gro and Shelter. No phytotoxicity of any kind was observed with the application of the micro-organism formulations and they could therefore be deemed suitable to be used for the treatment of maize seed. The micro-organism formulations containing fungal and bacterial biological control agents have the potential to be used in commercial maize production to increase vegetative and reproductive yields and reduce the severity of ear and stem rot in maize. / Agriculture Animal Health & Human Ecology / M.Sc. (Agriculture)

Thesis (PhD (Agric))--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: Biological control studies were conducted with isolates of entomopathogenic fungi Beauveria bassiana and Metarrhizium anisopliae from Ethiopia and South Africa against the spotted stem borer Chilo partellus. The study was conducted from April 2002 to April 2003, at the department of Entomology and Nematology, University of Stellenbosch, South Africa. The objectives were to screen these isolates for pathogenicity and to determine the susceptibility of different larval instars; to study the effect of temperature on fungal development and virulence; to investigate food consumption of fungus treated larvae; to determine compatibility of fungal isolates with insecticides; to study the effect of exposure methods and diets on larval mortality; and to evaluate promising isolates under greenhouse conditions using artificially infested maize plants. Four isolates of B. bassiana and six isolates of M. anisopliae were tested against second instar larvae. Of these isolates, B. bassiana (BB-01) and M. anisopliae (PPRC-4, PPRC-19, PPRC-61 and EE-01) were found to be highly pathogenic inducing 90 to 100 % mortality seven days after treatment. In subsequent assays, the fungal isolates were tested against third, fourth, fifth and sixth instar larvae. Second and sixth instar larvae were more susceptible to these isolates than third, fourth and fifth instar larvae. Conidial germination, radial growth and sporulation of the isolates PPRC-4, PPRC-19, PPRC-61, EE-01 and BB-01 were retarded at 15 and 35 0C. A suitable temperature range for the isolates was from 20 - 30 0C. At 25 and 30 0C the isolates induced 100 % mortality to second instar larvae within four to six days. Second and third instar C. partellus larvae were treated with the isolates PPRC-4 and BB-01, and daily consumption of maize leaf was measured. Treatment with the fungi was associated with a reduction in mean daily food consumption. In in-vitro studies, five concentrations (0.1 ppm, 1 ppm, 5 ppm, 10 ppm, and 100 ppm active ingredients) of the insecticides benfuracarb and endosulfan were tested with the isolates PPRC-4, PPRC-19, PPRC-16, EE-01 and BB-01. Increasing the concentration of the insecticides adversely affected germination, radial growth and sporulation of the isolates. In in-vivo studies combining the fungi, PPRC-4 and BB-01,with low concentrations (1 and 5 ppm a.i.), of the insecticides increased the mortality of third instar larvae from 65 to 100 %. Larvae sprayed directly with conidia, exposed to conidia treated leaves and dipped into conidial suspensions suffered high mortality of 98 to 100 %. Larvae exposed to treated leaves and larvae sprayed directly with conidia produced high numbers mycoses in cadavers. Exposure of larvae to treated leaves yielded high sporulation. At a low conidial concentration (1.25x107 conidia/ml), mycosis and sporulation were high. The optimum temperature was 20 0C for mycosis and 15 0C for sporulation. In greenhouse trails, a conidial suspension of 2 x 108 conidia/ml of the pathogenic isolates was sprayed on 3 to 4 week-old maize plants infested with 20 second instar larvae per plant. This resulted in suppression of foliar damage. Treatment with the fungi also reduced stem tunneling and deadheart. In addition, fungal treatment increased mean plant fresh and dry biomass compared to untreated control plants. In general, results from laboratory and greenhouse studies indicated that there was good potential for the use of these fungal isolates for controlling C. partellus larvae. / AFRIKAANSE OPSOMMING: Biologiese beheerstudies is uitgevoer met isolate van die insekpatogeniese swamme, Beauvaria bassiana en Metarrhizium anisopliae teen die gespikkelde stamboorder, Chilo partellus. Die doelwitte was om hierdie isolate te evalueer vir patogenesiteit; die vatbaarheid van verskillende larvale instars teenoor hulle te bepaal; die invloed van temperatuur op swamontwikkeling en virulensie te bepaal; die voedsel inname van swambehandelde larwes te ondersoek; die verenigbaarheid van die swamisolate met insektedoders te bepaal; die invloed van blootstellingsmetodes en diëte op larvale mortaliteit; en om belowende isolate in glashuisproewe te evalueer met gebruik van kunsmatig besmette mielieplante. Vier isolate van B. bassiana en ses isolate van M. anisopliae is teen tweede instar larwes getoets. Uit dié isolate is B. bassiana (BB-01) en M. anisopliae (PPRC-4, PPRC- 19, PPRC-16 en EE-01) as hoogs patogenies bevind. Hulle het 90 tot 100 % mortaliteit na sewe dae veroorsaak. In daaropvolgende essays, is die swamisolate teen derde, vierde, vyfde en sesde instar larwes getoets. Tweede en sesde instar larwes was gevoeliger vir die isolate as die derde, vierde en vyfde instar larwes. Spoorkeming, radiale groei en sporulasie van die isolate PPRC-4, PPRC-19, PPRC-61, EE-01 en BB-01, is by 15 en 35 0C vertraag. ‘n Aanvaarbare temperatuurreeks vir die isolate is vanaf 20 tot 30 0C. By 25 en 30 0C het die isolate 100 % mortaliteit teen tweede instar larwes binne vier tot ses dae geïndusseer. Tweede en derde instar C. partellus larwes is met die isolate PPRC-4 en BB-01 behandel en die daaglikse inname van mielieblare gemeet. Behandeling met die swamme is met ‘n afname in die gemiddelde voedselinname geassosieer In in-vitro studies is vyf konsentrasies (0.1 dpm, 1 dpm, 5 dpm, 10 dpm en 100 dpm aktiewe bestandele) van die insekdoders, benfuracarb en endosulfan getoets saam met die isolate PPRC-4, PPRC-19, PPRC-16, EE-01 en BB-01. ‘n Toename in die konsentrasie van die insekdoders het ontkieming, radiale groei en sporulasie van die isolate benadeel. In in-vitro studies het die kombinering van die die swamme, PPRC-4 en BB-01, met lae konsentrasies (1 en 5 dpm a.b.) van die insekdoders mortaliteit van derde instar larwes vanaf 65 tot 100 % laat toeneem. Larwes wat direk met spore gespuit is, aan behandelde blare blootgestel is en in spoorsuspensies gedoop is het tot hoë mortaliteit gelei, (98 tot 100 %). Blootstelling aan behandelde blare saam met ‘n lae konidiakonsentrasie, 1.25x106 spore/ml) en ‘n temperatuur van 15 tot 20 0C het tot hoë swammikose en sporulasie in kadawers gelei. In glashuisproewe, is ‘n spoorsuspensies van 2 x 108 spores/ml van die patogeniese isolate op 3 tot 4 weekoud mielieplante wat met 20 tweede instar larwes per plant besmet is gespuit. Dit het blaarskade onderdruk. Behandeling met die swamme het ook stamtonnels en dooiehart verminder. Boonop het swambehandeling die vars- en droë plantbiomassa laat toeneem in vergelyking met die onbehandelde kontrole plante. Oor die algemeen het resultate van laboratorium- en glashuisproewe getoon dat daar goeie potensiaal is vir die gebruik van hierdie swamisolate vir die beheer van C. partellus larwes.