In this study, we have demonstrated that inoculation of bean seeds with non-pathogenic binucleate Rhizoctonia (np-BNR) at sowing protected bean seedlings from infection of R. solani. Using quantitative real-time RT-PCR (QRT-PCR), transcript levels of defense genes encoding 1,3-beta-glucanase (GLUC), phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) in one-week old bean seedlings was monitored during np-BNR and R. solani interaction. The results revealed that protection effect of np-BNR correspond to a systemic suppression of these three defense genes' expression from significant higher level elicited by R. solani to the level of non-infected plants. This indicates that bio-protection by np-BNR isolates is not correlated to activation of these three defense genes' expression. Similar suppression was achieved for pre-colonization of bean seedlings with arbuscular mycorrhizal (AM) Glomus introradices on GLUC gene expression, although the AM fungus did not significantly reduce rot symptoms. Possible mechanisms implicated in down-regulation during plant-pathogen and np-BNR or AM interaction are discussed.

The suitability of sunflower (Heliantus annuus L.) as a host of Ophraella communa Le Sage (Coleoptera: Chrysomelidae) was evaluated under greenhouse and field conditions. Population dynamics of O. communa on sunflower and on ragweed (Ambrosia artemisiifolia L.) were determined using a life table approach. Sixty percent of O. communa females died during the first 30 days on sunflower while only 14% died on ragweed plants. Only 20% of fertile females laid eggs on sunflower plants as compared with 100% on ragweed plants. Fecundity, life duration, egg viability, and other biological parameters were significantly higher on ragweed plants than on sunflower plants. Ragweed is the main host plant for O. communa. Nevertheless, in specific situations (no-choice) adults and larvae of O. communa can damage sunflower plants. However, O. communa cannot complete its life cycle or increase its population on sunflower plants.

The Russian wheat aphid, Diuraphis noxia is one of the most destructive exotic invaders of South Africa, capable of reducing individual wheat plant yields by up to 90%. Entomopathogenic fungi are important natural mortality factors associated with this aphid in its Eurasian endemic habitats as well as in the United States and Canada; their impact often exceeding that of predators and parasitoids. The principal objectives of this study included (1) baseline characterization of the aphid-pathogenic flora associated with aphids from South Africa, with special reference to six common cereal aphids, (2) quantification of the comparative impact of the different fungal species on the cereal-aphid complex in three different wheat producing regions of South Africa, (3) field evaluation of the Hyphomycete Beauveria bassiana against D. noxia on resistant wheat, (4) screening of six fungicides for their potential usage in managing entomophthoralean epizootics within greenhouse rearings of the Russian wheat aphid, and (5) development and evaluation of a novel bioassay protocol for screening entomopathogenic Hyphomycetes against D. noxia. A total of nine species of fungi known to infect and kill aphid hosts were collected, including the six entomophthoraleans, Pandora neoaphidis, Conidiobolus thromboides, Conidiobolus obscurus, Entomophthora planchoniana, Conidiobolus coronatus and Neozygites fresenii, and three Hyphomycetes, Beauveria bassiana, Verticillium lecanii, and Paecilomyces farinosus. The former four entomophthoraleans are considered first reports from this country. For the first time, morphological characteristics of these nine South African-collected species are visually depicted and techniques for their isolation and in vitro culture discussed. Seven species of fungi were recorded from D. noxia, of which P. neoaphidis was the most important, causing up to 50% mortality during the late season under dryland conditions in the summer-rainfall region. Mycoses at epizootic levels, together with the large diversity of fungal species recorded from this host, indicated a high level of susceptibility to fungal infection. In contrast, infection of the oat aphid Rhopalosiphum padi remained < 5% despite favourable numbers of hosts and apparently suitable environmental conditions. This phenomenon strongly suggests some level of low susceptibility to fungal infection in this species. Under irrigated conditions m KwaZulu-Natal, the rose-gram aphid Metopolophium dirhodum was the predominant aphid but remained below economical injury levels. Field surveys revealed that this aphid was effectively targeted by P. neoaphidis and C. obscurus, and findings suggest that in some areas of South Africa entomophthoralean fungi effectively suppress this aphid, negating the need for insecticide applications. On average, ca. 61% control of D. noxia on resistant wheat was observed following an application of B. bassiana (5 x 10¹³ conidia per hectare) during the early flag-leaf stage of the wheat. Efficacy of B. bassiana applications on younger plants appeared to be influenced by the level of aphid activity, possibly explained by secondary pick-up of inoculum by D. noxia. These findings accentuate the importance of understanding the tritrophic relationship between host plant, pest and pathogen. The fungicides copper oxychloride, mancozeb + oxadixyl, captab + metalaxyl, bittertanol, iprodione, and mancozeb at a rate of 0.1% active ingredient moderately to strongly inhibited C. thromboides vegetative growth (mean inhibition 81.1 %). Mancozeb at concentrations of 10.0, 2.0, 1.25, 0.5, 0.08, and 0.016% was further evaluated in vitro. The fungus growth cut-off point, midway between 1.25 and 2.0% mancozeb, was calculated and a rate of 1.625% active ingredient per litre of water was used to decontaminate the fungus-infected D. noxia cultures. A novel bioassay protocol was developed, employing live host plants for rearing aphids post inoculation. Using this design against D. noxia, an average LC₅₀ estimate of 85 conidia per mm² for B. bassiana strain GHA was calculated. Control mortality was restricted to levels below 4%. The data indicated high precision due to an average coefficient of variation for slope of less than 20%, and an average chi-squared value of 5.46 ± 2.74 (n = 10 assays). The design will accommodate the use of cereal aphid species other than D. noxia, while live host plants will facilitate tritrophic studies on the effect of host-plant resistance on fungus-induced mortality of D. noxia. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2002.

The following body of research provides a detailed overview of the interactive effects of biocontrol agents and environmental factors and how these influence both the host plant and pathogen populations within hydroponic systems. Pythium and other zoosporic fungi are pathogens well suited to the aquatic environment of hydroponics. Motile zoospores facilitate rapid dispersal through fertigation water, resulting in Pythium becoming a yield reducing factor in most hydroponic systems and on most crops. With increasing trends away from pesticide use, biocontrol is becoming an ever more popular option. Unfortunately, much of our knowledge of biocontrol agents and their formulation can not be directly transferred to the widely differing environments of hydroponic systems. Paulitz (1997) was of the opinion that if biocontrol was to be successful anywhere, it would be in hydroponics. This is primarily due to the increased ability, in hydroponics, to control the growing environment and to differentiate between the requirements of the pathogen versus those of the host plant and biocontrol agent. Key environmental factors were identified as soil moisture, root zone temperature, form of nitrogen and pH. A review of the literature collated background information on the effects of biocontrol agents and environmental manipulation on plant growth and disease severity in hydroponic systems. A commercial formulation of Trichoderma (Eco-T(R1)) was used as the biocontrol agent in all trials. Dose responses in Pythium control and plant growth stimulation in lettuce were first determined using a horizontal trough system (closed system). In such systems optimum application rates were found to be lower than in field application (1.25x10[to the power of 5] spores/ml). This is probably because Trichoderma conidia are not lost from the system, but re-circulate until being transported into the root zone of a host plant. No significant growth stimulation was observed, although at high doses (5x10[to the power of 5] and 2.5x10[to the power of 5] spores/ml) a significant reduction in yield was recorded. Possible reasons for this growth inhibition are suggested and a new theory is proposed and investigated later in the thesis. In an open system of cucumber production (drip irrigated bag culture) no statistically significant results were initially obtained, however, general trends still showed the occurrence of positive biocontrol activity. The initial lack of significant results was mostly due to a poor knowledge of the horticulture of the crop and a lack of understanding of the epidemiology behind Trichoderma biocontrol activity. These pitfalls are highlighted and, in a repeat trial, were overcome. As a result it could be concluded that application rates in such systems are similar to those used in field applications. Management of soil moisture within artificial growing media can aid in the control of Pythium induced reductions in yield. A vertical hydroponic system was used to determine the interactive effects of soil moisture and Trichoderma. This system was used because it allowed for separate irrigation regimes at all 36 stations, controlled by a programmable logic controller (PLC). With lettuce plants receiving optimum irrigation levels, no significant reduction in yield was observed when inoculated with Pythium. However, after Pythium inoculation, stresses related to over- or under-watering caused significant yield losses. In both cases, Trichoderma overcame these negative effects and achieved significant levels of disease control, especially under higher soil moisture levels. Growth stimulation responses were also seen to increase with increasing soil moisture. Similar results were obtained from strawberry trials. These results show that Pythium control is best achieved through the integration of Trichoderma at optimum soil moisture. However, where soil moisture is above or below optimum, Trichoderma serves to minimize the negative effects of Pythium, providing a buffering capacity against the effects of poor soil moisture management. Pythium, root zone temperature and form of nitrogen interact significantly. In greenhouse trials using horizontal mini troughs with facilities for heating or cooling recirculating water, nitrate fertilizer treatments resulted in statistically significant results. Lettuce growth was highest at 12°C, although no significant differences in yield were observed between 12-24°C. Pythium was effective in causing disease over the same temperature range. Pythium inoculation did not result in yield reduction at 6 and 30°C. Trichoderma showed a slight competitive advantage under cooler temperatures (i.e., 12 degrees C), although significant biocontrol occurred over the 12-24 degrees C range. Ammonium fertilizer trials did not generate statistically significant data. This is possibly due to complex interactions between root temperature, ammonium uptake, and competitive exclusion of nitrification bacteria by Trichoderma. These interactions are difficult to replicate over time and are probably influenced by air temperature and available light which are difficult to keep constant over time in the system used. However, the data did lead to the first clues regarding the effects of Trichoderma on nitrogen cycling as plants grown with a high level of ammonium at high temperatures were seen to suffer more from ammonium toxicity when high levels of Trichoderma were added. In further trials, conducted in the recirculating horizontal mini trough system, it was determined that Trichoderma applications resulted in an increase in the percentage ammonium nitrogen in both the re-circulating solution and the growing medium. This was a dose-related response, with the percentage ammonium nitrogen increasing with increasing levels of Trichoderma application. At the same time an increase in ammonium in the root tissue was observed, corresponding with a decrease in leaf nitrate levels and an increase in levels of Cu, Na, Fe and P in leaf tissue. In independent pot trials, populations of nitrifying bacteria in the rhizosphere were also seen to decrease with increasing Trichoderma application rates. This led to the conclusion that the increase in ammonium concentration was as a result of decreased nitrification activity due to the competitive exclusion of nitrifying bacteria by Trichoderma. The possibility that Trichoderma functions as a mycorrhizal fungus and so increases the availability of ammonium for plant uptake is not discarded and it is thought that both mechanisms probably contribute. Water pH provides the most powerful tool for enhancing biocontrol of Pythium by Trichoderma. Trichoderma shows a preference for more acidic pHs while Pythium prefers pHs between 6.0 and 7.0. In vitro tests showed that Trichoderma achieved greater control of Pythium at pH 5.0, while achieving no control at pH 8.0. In greenhouse trials with the recirculating horizontal mini trough system, yield losses resulting from Pythium inoculation were greatest at pH 6.0 and 7.0, with no significant reduction in yield at pH 4.0. Biocontrol activity showed an inverse response with greatest biocontrol at pH 5.0. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2003.

The two-spotted spider mite (TSM), Tetranychus urticae Koch, is an important pest of many greenhouse and field crops worldwide. The development of resistance in TSM populations to chemical acaricides, allied with public health concerns about pesticide residues, has motivated the search for alternative control measures to suppress the pest. Hyphomycetous fungi are promising agents for mite control and the fungus Beauveria bassiana (Bb) (Balsamo) Vuillemin was investigated in this study as a biocontrol agent. The principal objectives of this study comprised: a) screening Bb strains for their pathogenicity against T. urticae; b) testing the effect of adjuvants on the efficacy of Bb; c) studying the effect of plant type on persistence of Bb and the efficacy of control of Bb against T. urticae; d) evaluating the field efficacy of Bb applications against T. urticae; e) testing the compatibility of Bb with selected fungicides; and f) assessing the synergy between Bb and soluble silicon for T. urticae control. Screening bioassays of sixty-two strains of Bb identified the two most effective strains, PPRI 7315 (R289) and PPRI 7861 (R444), that caused mortality levels of more than 80% of adult mites at 9 d post-inoculation with 2 × 108 conidia ml-1. These strains performed significantly better than the Bb commercial strain PPRI 5339, in laboratory bioassays. The two strains also attacked mite eggs, causing 53.4% and 55.5% reduction in egg hatchability at 2 × 108 conidia ml-1 respectively. However, PPRI 7861 showed relatively higher production of conidia in culture and was, therefore, selected for further trials under greenhouse and field conditions. Greenhouse evaluations of the effects of two adjuvants (Break-thru® and a paraffin oil-based emulsion) on efficacy of Bb demonstrated a higher efficacy of the biocontrol agent (BCA) when it was applied with Break-thru® or the oil solution than with water alone. Moreover, Bb conidia applied in Break-thru® solution resulted in greater control of TSM than conidia applied in the mineral oil. There was also a dose-response effect and the control of TSM by Bb increased when the concentration of conidia was increased. The control of TSM by Bb in beans (Phaseolus vulgaris L), cucumber (Cucumis sativus L.), eggplant (Solanum melongena L.), maize (Zea mays L.) and tomato (Solanum lycopersicum L.) was tested in greenhouse trials. On these crops, the persistence of conidia declined over time. The rate of decline was significantly higher on maize. However, TSM mortality was positively correlated with the amount of conidia deposited on leaves immediately after spraying, rather than their persistence over time. Higher levels of mortality of TSM due to Bb application were observed on beans, cucumber and eggplants, suggesting that the type of crop must be taken into consideration when Bb is applied as a BCA. Field efficacy of Bb against mites was evaluated in two trials on eggplants. Based on assessment of population densities of mites and leaf damage assessments; both trials showed that the strain PPRI 7861 controlled TSM in the field. Two commonly used fungicides, azoxystrobin and flutriafol, were investigated in vitro tests on culture medium and laboratory bioassays on detached bean leaves (Phaseolus vulgaris L.) for their effects on Bb. Azoxystrobin (a strobilurin) was less harmful to Bb while flutriafol was found to be inhibitory. Another important finding of this study was the substantial enhancement of Bb efficacy by soluble silicon. When Bb was combined with soluble Si, the control of TSM was better than when either of the two products was applied alone. Moreover, application of soluble Si as a plant fertilizer in hydroponic water nutrient increased accumulation of peroxidase, polyphenoloxidase and phenylalanine ammonia-lyase enzymes in leaves of plants infested with TSM. Increased activity of these defense enzymes in leaves deters feeding behaviour of mites. We suggested that feeding stress renders them susceptible to Bb infection, which would explain the synergy observed between the two agents. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Bracken fern (Pteridium aquilinum [L.] Kuhn) is an indigenous invasive plant and it is known to have a negative impact on biodiversity. This research focuses on infestations of bracken fern in two areas within the uKhahlamba Drakensberg Park World Heritage Site; the Royal Natal National Park and the Rugged Glen Nature Reserve. Prior change detection research on bracken fern were constrained due to the low resolution satellite imagery and the inability of hard classification techniques to account for the mixtures of land cover types that occur within pixels of low resolution imagery. To overcome these constraints this research applied the fuzzy image classification technique to multispectral digital aerial imagery of 0.5 m spatial resolution. Multi date imagery used for image classification was captured in the mid-winter of 2009 and mid-spring of 2011. Thereafter post-classification change detection analysis was conducted using the fuzzy classified images. The classified images were verified using ground truth surveys. The 2009 and 2011 fuzzy classified images produced overall accuracies of 81.4% and 94.4% with Kappa coefficients of 0.63 and 0.89 respectively. This research found that the distinct seasonal development pattern of bracken fern and the time of year imagery were captured were significant factors in its detection. As bracken fern was found to be more spectrally distinct in spring as compared to winter, due to the plant growth of bracken fern, grass and other shrubbery. These classified images were used in post-classification change detection analysis which revealed that the bracken fern infestation in the Royal Natal National Park and Rugged Glen Nature Reserve had increased at a rate of 24 % and 27 % per annum respectively. This showed that bracken fern is spreading in the Royal Natal National Park and Rugged Glen Nature Reserve, as expected. Fire regimes, slope and aspect were found as factors that could be promoting the spread of bracken fern, 67.5 % and 75 % of the bracken fern infestation in the Park and Reserve respectively, occurred in areas that were burnt by fire regimes and have gentle to moderately gentle slopes facing east, south east and south. / Thesis (M.Sc.Sur.)-University of KwaZulu-Natal, Durban, 2013.

The Labiatae is a large family that occurs worldwide and have species that are adapted to almost all habitats and altitudes. Plectranthus is in this family. Plectranthus species are beautiful South African shrubs. The genus Plectranthus belongs to subfamily Nepetoideae of tribe Ocimeae. The test microorganisms were chosen carefully as each one belonged to a different taxonomic group of fungi and bacteria. Biologically active mono- and sesquiterpenoids are frequently found in many species of Plectranthus but there are little published data that directly link the presence of specific compounds in a species with the traditional uses of that species. Various Plectranthus spp. were collected and dried, followed by chemical extraction using various solvents. Dichloromethane extracts of P. fruticosus and P. ecklonii were screened for antibacterial and antifungal activities using the agar well and trench diffusion methods. It was found that both methods produced inconsistent results. The trench method required a bigger volume of plant extract to be filled into the well, hence, better biological activity was observed with that method. The well method required a smaller volume therefore poor activity was noted with this assay. The size of inhibition zones are dosage dependent. Overall, both plant extracts exhibited antibacterial but no antifungal properties. The pure compound (1), 11-Hydroxy-2-(4-hydroxybenzoyl)-5,7,9(11),13-abietatetraen-12-one, isolated from P. ecklonii was found to be the same as compound (10) which was isolated from P. lucidus. P. hadiensis was extracted using dichloromethane and hexane. The dichloromethane extract proved to contain much higher biological activity than the hexane extract. Three pure compounds, identified as diterpenes, were isolated from the crude dichloromethane extract of P. hadiensis. 6,7-Dihydroxyroyleanone-6,7,12-trihydroxy-8,12-abietadiene-ll,14-dione (2) and 7(alpha)-formoxy-6(beta)-hydroxyroyleanone (3) exhibited good antibacterial and antifungal activity but not against all the test organisms. The remaining pure compound, 7(alpha)-acetoxy-6(beta)hydroxyroyleanone (4), exerted good antifungal activity. This was measured by the inhibition zone which measured up to 14mm when compound 4 was tested against S. sclerotiorum. When testing the hexane extract against the Bacillus formulations, the pellets that were suspended once in Ringer's solution produced bigger inhibition zones than the pellets that were suspended twice. This could be due to bacterial cells washing out of the suspension. The dichloromethane extract of P. praetermissus proved to be very active against X campestris, producing an inhibition zone of 8 - 20mm. Two pure compounds were isolated from the crude extract and identified as diterpenes. Compound 5, 20(10--> 5)-abeo1( 10),6,8,11,13-abietapentaene-11,12,16-triol, and compound 6, 11,12,15-trihydroxy-20( 10-->5)-abeo-abieta-1-(10),6,8,11,13-pentaene are both known compounds which have previously been isolated from Salvia apiana. P. cilatus was extracted with chloroform and tested against various microorganisms for antifungal and antibacterial activities. It showed poor biological activity overall, except against S. sclerotiorum. The crude dichloromethane extract of P. zuluensis exhibited good antibacterial activity, which was limited to the Gram negative test organism. The extract produced an inhibition zone of 10-12mm when tested against X campestris. Pure compound 7, 2-hydroxy-4,6dimethoxyacetophenone, exerted excellent inhibition against B. subtilis and S. sclerotiorum. Neither compound 8, 1,2,4-trimethoxy-5-(2-propenyl)-benzene, nor compound 7, inhibited Candida spp., F. oxysporum and R. solani. Two diterpenes were isolated from the aerial plant parts of P. lucidus with dichloromethane and their structures elucidated by spectroscopic means. The pure compound 9, 11-hydroxy19-( 3-methyl-2-butenoyl)-5,7,9(11), 13-abietatetraen-12-one, showed moderate antifungal activity whereas compound 10, 11-hydroxy-2-(4-hroxybenzoyl)-5,7,9(11),13-abietatetraen12- one, showed high antifungal activity against R. solani, S. sclerotiorum and F. oxysporum. The crude and the pure compounds (formerly isolated from P. parviflorus) showed inhibition against X campestris. The dichloromethane extracts of P. purpuratus subsp. purpuratus and P. purpuratus subsp. tongaensis exhibit similar levels of biological activity when tested against the same test organisms. Poor antibacterial activity was noted with both extracts. However, excellent antifungal activity was depicted when both plant extracts were tested against F. oxysporum, R. solani and S. sclerotiorum. However, the highest biological activity was noted by R. solani which was totally inhibited by both dichloromethane extracts. The pure compound (11) isolated from P. purpuratus subsp. purpuratus was found to have the same chemical structure as compound (9) previously isolated from P. lucidus. The bioautography assay was used to detect and activity-guide the fractionation of antimicrobial compounds from all the Plectranthus spp. tested. The TLC fingerprint showed a zone of clearing around the lower bands of P. fruticosus and P. ecklonii when the plate was sprayed with a suspension of B. subtilis. This result is consistent with the agar well diffusion method. Clear zones were also noted on some bands of the extracts of P. zuluensis, P. ciliatus, P. hadiensis and P. praetermussis. Clear zones indicate inhibition of growth. Other plant extracts tested for biological activity were from the family Lamiaceae, however, not of the genus Plectranthus. Persicaria senegalensis, Pycnostachys reticulata and Ficus sur possessed moderate biological activity overall. It is interesting to note that P. senegalensis and F. sur exert high biological activity against Candida spp. This could be useful as herbal remedies for yeast infections. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

The following studies were done to identify factors that influence the virulence of Xenorhabdus nematophilus and the interactions of the bacterium with the nonself defence systems of nonimmune Galleria mellonella. Isolates of X. nematophilus that are qualitatively similar in biochemical properties differed significantly in virulence for G. mellonella larvae. The production of enzymes such as proteases could not account for differences in the virulence of the isolates. / Growth conditions, influenced the growth rate and the interactions of the bacterium with nonimmune G. mellonella larvae. In general, X. nematophilus cells grown under aerobic conditions were more susceptible to the nonself defences of G. mellonella larvae than those grown under less than ideal conditions, resulting in increased clearance of the bacteria from the hemolymph (blood) of the insects. Clearance of the bacteria from the hemolymph of the insect was positively correlated with culture condition, culture age, and attachment to insect hemocytes in vitro. / Isolates of X. nematophilus produced flagella and fimbriae when grown under microaerobic and aerobic conditions. The type of fimbriae produced was influenced by culture conditions. The injection of both flagella and fimbriae in picogram quantities into nonimmune G. mellonella caused an increase in total hemocyte counts within these insect larvae. The injection of fimbrial and flagellar antigens into G. mellonella larvae caused changes in the hemocyte types found in circulation in the insect's hemolymph. (Abstract shortened by UMI.)

At present, compared to our scientific knowledge of how natural biosystems function, we have practically no knowledge of how to engineer biosystems, i.e., how to design, build, repair, maintain, operate, and modify them in a rational and knowledgeable manner. Thus, the EcoCyborg research project was established with the basic intent of making a contribution to the science and engineering of biosystems. / This thesis covers three important arms of study within the overall framework of the EcoCyborg project. The first area is the development and implementation of a workbench for simulation based on composite models. The workbench was designed to accommodate easy integration, synchronization and communication of a number of independent modules. It was developed for use under OS/2 Warp, an operating system that supports multi-process simulation. The functionality of the workbench was tested with a composite model of an EcoCyborg. This composite model contained submodels of an ecosystem, two control systems, and several forcing functions. / The second area is the development and implementation of control systems for biosystems. The control system was intended to guide an ecosystem so that its behavior would best achieve the goals of the controllers which, in this case, were related to the populations of the ecosystem's species. The major part of the work on the control systems was focused on the Pavlovian controller. It mainly covered the investigation of the possibilities to develop and to implement a physiological-level Pavlovian controller. The thesis also presents results from a preliminary investigation on the design and implementation of a cognitive controller. From this, a relatively simple cognitive controller was developed to complement the action of the Pavlovian controller. / The third area is the experimental use of the workbench, and of the Pavlovian and cognitive controllers. For this, ecosystems having various constitutions and their forcing functions were modeled and used in the simulation. Pavlovian controllers with on-off, proportional and proportional plus integral mechanisms were used to control the populations. The cognitive controller was used to maintain minimum population of the various species in the ecosystem. / In general, the workbench that was developed was found suitable to accommodate the simulation of an EcoCyborg composite model. The tools that ware developed for constructing and implementing the Pavlovian controller were tested and found practical and effective for use in constructing Pavlovian controllers of various configurations. The results of the work on the cognitive controller provided useful information for its further development.

Components of successful pest management programs must be complementary and not antagonistic. This project examined interactions between natural enemies of the black cutworm, Agrotis ipsilon (Hufnagel), an important turfgrass pest, and host plant resistance by endophytic grass. Agrotis ipsilon nucleopolyhedrovirus (AgipMNPV) was examined as a bio-insecticide for controlling A. ipsilon in turfgrass. Fresh (1-week-old) AgipMNPV residues killed 76−86% of neonates hatching from eggs on golf course tees, however, residual control of implanted larvae lasted no more than a few weeks. Combinations of AgipMNPV with adjuvants, such as optical brightener and lignin, failed to accelerate or extend efficacy of the virus. AgipMNPV seems better suited for targeted control of early instars than for season-long control. Several applications per growing season would likely be needed to maintain high enough titers on turfgrass to effectively control cutworms. The addition of a chitin synthesis inhibiting turfgrass fungicide failed to synergize AgipMNPV infectivity to A. ipsilon. Choice tests revealed the fungicide residues to be a mild feeding deterrent, the likely cause of slightly reduced mortality from virus infection seen in field trials. Combination applications in turfgrass might interfere with larval ingestion of a lethal virus dose, resulting in prolonged feeding in the field. I examined how feeding on perennial ryegrass (Lolium perenne) with or without Neotyphodium lolii, its alkaloid-producing fungal endophyte, affects susceptibility of A. ipsilon to AgipMNPV. Feeding on endophytic grass neither compromises nor synergizes infectivity of AgipMNPV in the cutworm midgut. However, reduced consumption or avoidance of less-palatable endophytic grass could decrease ingestion of virus and rates of subsequent mortality in the field. Host feeding on endophytic grass had differing effects on the tachinid fly, Linnaemya comta, a fast-developing solitary parasitoid, and the encyrtid wasp, Copidosoma bakeri, a slow-developing gregarious parasitoid. L. comta development did not appear to be affected when its host fed on endophytic grass; in contrast, C. bakeri suffered negative fitness effects. These results suggest that parasitoid life strategy and taxonomy play a role in endophyte mediated tritrophic interactions.