Rozprawy doktorskie na temat „Biological pest control”

Includes bibliographical references (leaves 168-198) The horehound plume moth (Wheeleria spilodactylus Curits), an agent introduced to control the invasive weed horehound (Murrubium vulgare L.), was used as a model system to investigate factors believed to influence the successful establishment of an introduced natural enemy. Retrospectively tests the use of generic population viability analysis and decision making tools for determining optimal release strategies for the horehound plume moth in South Australia and to compare outcomes with the emprical data collected during the course of this project

The activity of Bacillus brevis Nagano and the antibiotic it produces, gramicidin S, against the polyphagous plant pathogen, Botrytis cinerea was studied in vitro. Both germination and growth of B. cinerea were sensitive to the antibiotic although germination was the more sensitive. Sensitivity towards the antibiotic varied with cultural conditions. Treatment of B. cinerea conidia with high levels of gramicidin S caused massive release of cellular ATP indicating membrane disrupton as a primary mode of action. Activity of gramicidin S against conidia of B. cinerea was much lower when applied to leaves of Chinese cabbage compared to in vitro. This reduced activity was thought to be due to the binding of gramicidin S to ethanol soluble component(s) of the leaf surface. Gramicidin S also bound strongly to soil, especially the organic moiety. This gave the antibiotic low mobility and low bioavailability within soil. Methods were developed for the selective retrieval and enumeration of both B.brevis and B. cinerea from plant and soil material. Field trials showed that treatment of Chinese cabbage with B.brevis Nagano was as effective as standard fungicide treatment with iprodione in controlling grey mould infection in two successive field trials. This observation was not expected from the in planta studies concerned with antibiosis since it was indicated that gramicidin S had low activity on leaf surfaces. Observations of leaf wetness duration (LWD) indicated that B.brevis Nagano preparations contained a biosurfactant within the supernatant which altered the wettability of leaves. In greenhouse trials leaves dried 80 percent quicker after irrigation when treated with B. brevis Nagano culture compared to untreated control leaves.

Dasineura sp. is a gall forming midge that was introduced into South Africa for the biocontrol of the Australian myrtle, Leptospermum laevigatum. It causes galls on both the vegetative and reproductive buds of the plant. Although Dasineura sp. was initially regarded as a potentially successful agent, galling up to 99 percent of the buds of the host plant, it has been preyed on by native opportunistic mites, which caused a decline in the performance of the midge as a biocontrol agent of L. laevigatum. This raised a concern about whether this fly will be able to perform effectively in the presence of its new natural enemies. Therefore, the objectives of this study were to: 1) ascertain whether mite abundance has seasonal variations; 2) determine if plant density and plant size have an effect on midge predation by the mites; and 3) determine if midge predation varies in different locations. The study was conducted at three sites in the Hermanus area, Western Cape Province. Every three weeks for thirteen months, galls were collected and dissected so as to count and record the numbers of midge larvae, pupae, adults and mites that were found. Data collected showed that predation varied with season, and the mites were scarce during the flowering season. Predation also varied among the study sites and plant density had an effect on midge predation. Midges in smaller plants (saplings) were more vulnerable to predation than those in the bigger plants (plants from isolates and thickets). It was concluded that although mites have an effect on midge populations, they do not prevent their establishment on the plant. Therefore, a survey should be done in two to three years time to check if the midges are still persisting on the plant, vi and recommendations are that a new agent should be released to supplement the midges.

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.

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.

In agroecosystems, conservation biological control is often constrained by low availability of pollen and nectar. Although floral resources can be integrated into agricultural systems, this usually requires sacrifice of productive land. This thesis describes experiments which test the influence of floral resources provided by species which have marketable value in their own right as food or medicinal crops. Floralresource strips comprising such species were intercropped with Brassica crops at plot, field and commercial scales in replicated field experiments. The abundance and distribution of key Brassica crop pests a,nd their natural enemies in the presence of floral resources was examined. Parasitoids displayed distinct flower species preferences; Fagopyrum esculentum significantly increased parasitoid abundance. Parasitoids also responded to distance; their abundance significantly declined with increasing distance from floral strips, especially in F. esculentum, and parasitism levels declined significantly after 8 m. Borago offtcinalis also proved attractive to parasitoids and Tanacetum parthenium to Syrphidae. In addition, F. esculentum and B. offtcinalis were found to suppress the second generation of Pierid larvae at field scale. This evidence suggests that several flower species could be intercropped within a Brassica crop to promote biological pest control.

Invasive species are alien to the ecosystem under consideration and cause economic or environmental damage or harm to human health. Two alien insects that fit this description are the brown marmorated stink bug, Halyomorpha halys and the spotted lanternfly, Lycorma delicatula. Both invaders are polyphagous pests that feed on a myriad of plant species and inflict severe crop losses. As sustainable control methods depend on the accurate monitoring of species’ invasion and involve the use of natural enemies, we addressed these two facets by exploring novel monitoring techniques and deciphering host-parasitoid interactions for improved integrated pest management. Thus, we adopted ‘BugMap’, a citizen science initiative that enables students, farmers and everyday citizens to report sightings of H. halys from Italy, with emphasis on Trentino-Alto Adige. Aside from fostering citizen participation in scientific endeavors and the enhanced literacy that ensues, BugMap helped uncover the invasion dynamics of H. halys and forecast its potential distribution in Trentino, all while coordinating technical monitoring and informing management strategies. The most promising agent currently under study for the classical biological control of H. halys is the Asian egg parasitoid Trissolcus japonicus. To assess the wasp’s potential non-target impacts, we investigated its foraging behavior in response to chemical traces ‘footprints’ deposited by its main host H. halys and by a suboptimal predatory species, the spined soldier bug, Podisus maculiventris. Wasps exhibited a ‘motivated searching’ when in contact with footprints originating from both species. However, T. japonicus arrestment was significantly stronger in response to H. halys footprints, compared with P. maculiventris, implying the presence of underlying chemical cues that shape its natural preferences. A series of GC-MS chemical analyses revealed that n-tridecane and (E)-2-decenal were more abundant in H. halys footprints and are probably the key components utilized by the wasp for short range host location. The function of the aforementioned compounds was studied, n-tridecane acted as an arrestant, prolonging T. japonicus residence time, whereas (E)-2-decenal fulfilled its presumed defensive role and repelled the wasp. These results shed new light on the chemical ecology of T. japonicus and help expand the understanding of parasitoid foraging and its implications for classical biological control. Moving to the other invader L. delicatula, an egg parasitoid Anastatus orientalis was reported attacking it at high rates in its native range in Eastern Asia and may play a key role in reducing its populations there. A series of bioassays revealed that wasps responded to footprints deposited by L. delicatula gravid females by initiating a strong searching behavior. Moreover, A. orientalis preferred to oviposit in egg masses with intact oothecae, suggesting that the host’s egg covering functions as a trigger for A. orientalis probing and oviposition. Thus, A. orientalis not only overcomes, but also reverses an important line of host structural defense for its own fitness gains. This dissertation discusses the benefits of combining citizen science with traditional monitoring, and the usefulness of decoding host-parasitoid interactions to design more efficacious management strategies of invasive insect pests.

Yellow toadflax, Linaria vulgaris (L.) Mill. (Plantaginaceae), is a non-native invasive plant. Rhinusa pilosa Germar (Coleoptera: Curculionidae) is a proposed biocontrol agent. Gall development by R. pilosa was described using histological methods and compared between plant populations from native and introduced ranges. Key stages of oviposition were isolated histologically to determine their importance in gall induction. Rhinusa pilosa galled and developed on four geographically distinct Canadian populations in a pre-release quarantine study. Low agent densities only negatively affected one population. High densities of R. pilosa reduced potential reproductive output and plant biomass. Conducting detailed investigations into the biology, impact, and development of R. pilosa on populations from invasive and native ranges may help predict the efficacy of R. pilosa in the field if approved for release and.goes beyond current pre-release testing requirements.
ix, 168 leaves : ill. (chiefly col.) ; 29 cm

Doutoramento em Engenharia Florestal e dos Recursos Naturais - Instituto Superior de Agronomia
The Australian weevil Gonipterus platensis (Marelli) (Coleoptera: Curculionidae), commonly known as the Eucalyptus snout-beetle, is one of the main pests of eucalypts. Because this is a non-native species, classical biological control with natural enemies from its region of origin should be a viable control strategy. The introduction of the Australian parasitoid Anaphes nitens (Girault) (Hymenoptera: Mymaridae) has been the main method to control the pest worldwide. However, this natural enemy is not completely effective in reducing G. platensis populations and does not avoid the occurrence of damage in several regions. Therefore, it is important to identify effective control alternatives for these areas. In this work, the economic impact of the pest and the benefit of biological control with A. nitens over the last 20 years were assessed, using Portugal as a case study. The results of the economic analysis showed that, without biological control, the losses caused by G. platensis would be at least four times higher than those occurring with partial control by A. nitens. Given the insufficient efficacy of A. nitens, the existence of other natural enemies in Australia that could be used in a classical biological control programme was evaluated. From a set of eight natural enemies identified in Tasmania, the egg parasitoid Anaphes inexpectatus Huber and Prinsloo (Hymenoptera: Mymaridae) was selected for further studies. Laboratory studies comparing the biology of A. inexpectatus and A. nitens at different temperatures, a competition study between these two species, and a risk analysis for the introduction of A. inexpectatus in the Iberian Peninsula were carried out. Overall results suggest that A. inexpectatus might complement parasitism by A. nitens under field conditions without non-target effects on native fauna
N/A

Thesis (MEng) -- Stellenbosch University, 2014.
ENGLISH ABSTRACT: As overpopulation threatens the world’s ability to feed itself, food has become an invaluable resource. Unfortunately, almost a third of the food produced for human consumption is lost annually. Pests including insects, phytopathogens and weeds are responsible for more than a third of the annual major crop losses suffered around the world. The majority of current post-harvest control strategies employ synthetic agents. These compounds, however, have been found to be detrimental to the environment as well as human health, which has led researchers to investigate alternative strategies. Biocontrol agents are environmentally compatible, have a lower toxicity and are biodegradable, making them an attractive alternative to the synthetic control agents. The lipopeptides produced by Bacillus spp. in particular, have shown great potential as biocontrol agents against various post-harvest phytopathogens. Most biocontrol strategies apply the biocontrol organism, for example Bacillus, directly, whereas this study focused on the use of the lipopeptide itself as an antifungal agent. This is advantageous as the lipopeptides are less sensitive to their surroundings, such as temperature and pH, compared to living organisms, allowing for the production of a standardized product. This study investigated the production of the Bacillus lipopeptides surfactin, fengycin and iturin under controlled batch conditions. Parameters increasing lipopeptide production were quantified, focussing on antifungal lipopeptides (iturin and fengycin), and lipopeptide production was optimized. Experiments were performed in a fully instrumented 1.3 L bench-top bioreactor and lipopeptide analyses were performed via high pressure liquid chromatography (HPLC) and liquid chromatography-mass spectroscopy (LC-MS). After screening four Bacillus spp., Bacillus amyloliquefaciens DSM 23117 was found to be the best antifungal candidate. This was based on it outperforming other candidates in terms of maximum antifungals produced, Yp/x,antifungals (yield per cells), and antifungal productivity. Nitrate, in the form of NH4NO3, was critical for lipopeptide production and an optimum concentration was observed above which the CDW (cell dry weight) no longer increased significantly and both μmax (maximum specific growth rate, h-1) and lipopeptide production decreased. For μmax, the optimum NH3NO4 concentration was 10 g/L and for lipopeptides it was 8 g/L. At these respective NH4NO3 concentrations μmax = 0.58 (h-1), the maximum antifungals (fengycin and iturin) were 285.7 mAU*min and the maximum surfactin concentration was 302 mg/L. The lipopeptides produced by B. amyloliquefaciens, the antifungals (fengycin and iturin) and surfactin, are secondary metabolites, regardless of the optimization treatment, i.e. increased NH4NO3 concentrations. Using 30% enriched air extended the nitrate utilization period, suggesting that when increasing supply concentration, more oxygen was available to act as electron acceptors, allowing nitrate to be used for lipopeptide production. The number of iturin and fengycin homologues generally increased with an increase in nitrate concentration. This suggested that process conditions, such as nitrate concentration, can be used to manipulate homologue ratios, allowing for the possibility to tailor-make biocontrol-agent upstream, during the production process, and possibly increase the efficacy of the biocontrol strategy. The lipopeptides produced by B. amyloliquefaciens showed complete inhibition against Botryotinia fuckeliana and diminished the growth capabilities of Botrytis cinerea. No inhibition was observed against Penicillium digitatum. These results indicate potential of the biocontrol strategy, although scale-up and fed-batch studies are recommended, especially when considering commercial implementation. Studies regarding the lipopeptide application method, i.e. a single application or multiple applications, should also be investigated as this will influence the efficacy of the lipopeptides against the target organisms.
AFRIKKANSE OPSOMMING: Met oorbevolking wat die wêreld se vermoë om die groeiende bevolking te onderhou belemmer, het dit noodsaaklik geword om huidige voedselbronne te beskerm. Daar word beraam dat een derde van die voedsel wat wêreldwyd geproduseer word vir menslike verbruik verlore gaan elke jaar. Verder is insekte, plantpatogene en onkruide verantwoordelik vir meer as ‘n derde van die verliese rakend jaarlikse oeste. Meeste bestaande na-oes beheermetodes maak gebruik van sintetiese stowwe. Ongelukkig kan hierdie verbindings nadelig wees vir die omgewing sowel as menlike gesondheid. Navorsers het hulsef dus toespits daarop om alternatiewe beheermetodes te ondersoek. Bio-beheermetodes is omgewingsvriendelik sowel as bio-afbreekbaar, wat hulle ideale alternatiewe maak vir die sintetiese stowwe. Bacillus spp. lipopeptiede het veral hoë potensiaal getoon as bio-beheermiddels teen verskeie na-oes plantsiektes. Meeste bio-beheermetodes wend die biobeheer organisme, soos Bacillus, direk aan, waar hierdie studie op die gebruik van lipopeptiede as ‘n beheermiddel gefokus het. Die voordeel is dat lipopeptidiede minder sensitief is vir hul omgewings, soos temperatuur en pH, i.v.m. organismes en die moontlikheid bied van ‘n gestandardiseerde produk. Hierdie studie het die produksie van spesifieke Bacillus lipopeptide, naamlik surfactin, fengycin en iturin, onder beheerde lottoestande ondersoek. Parameters wat lipopeptied produksie verhoog is gekwantifiseer, spesifiek antifungiese middels (iturin en fengycin) en lipopeptied produksie is geoptimeer. Eksperimente is uitgevoer in ‘n 1.3 L bioreaktor en lipopeptiedanaliese is met behulp van hoë druk vloeistof chromatografie en vloeistofchromatografie-massa spektroskopie uitgevoer. Van die vier moontlike Bacillus spp., was Bacillus amyloliquefaciens DSM 23117 die mees belowende antifungus-produserende kandidaat. Dit het beter resultate gelewer in terme van maksimale antifungiese produksie, Yp/x,antifungies (opbrengs per sel) asook antifungiese produktiwiteit. Nitraat, in hierdie geval NH4NO3, was noodsaaklik vir lipopeptied produksie en ‘n optimale konsentrasie is waargeneem waarbo die seldigtheid nie meer beduidend toegeneem het nie en beide die μmax (maksimale spesifieke groei tempo, h-1) en lipopeptied produksie afgeneem het. Die optimale NH4NO3 konsentrasie vir μmax was 10 g/L en vir lipopeptiedproduksie was 8 g/L. By 10 g/L NH4NO3 was μmax = 0.58 (h-1) en by 8 g/L was die maksimale antifungiese produksie (fengycin en iturin) 285.7 mAU*min en die maksimale surfactin produksie 302 mg/L onderskeidelik. Die lipopeptide, die antifungiese middels (fengycin en iturin) en surfactin, geproduseer deur B. amyloliquefaciens is sekondêre metaboliete, ongeag van die optimerings-behandelinge wat toegepas word, soos ‘n verhoging in NH4NO3 konsentrasie. Die gebruik van 30% verrykte suurstof het die nitraat verbruikingsperiode verleng, wat voorgestel het dat met die verryking, meer suurstof beskikbaar was om te dien as finale elektron ontvanger en sodoende die nitraat beskikbaar te stel vir lipopeptied produksie. Iturin en fengycin homoloë, oor die algemeen, het toegeneem soos wat die nitraat konsentrasie verhoog is. Hierdie resultate dui daarop dat prosestoestande, soos nitraat konsentrasie, gebruik kan word om die verhouding waarin lipopeptied homoloë geproduseer word te manipuleer. Hierdie resultate dui op die potensiaal vir die stroomop produksie van ‘n unieke bio-beheermiddel, wat die effektiwiteit van die bio-beheermetode moontlik sal verhoog. Die geproduseerde lipopeptiede het totale inhibisie getoon teen Botryotinia fuckeliana en ook fungiese aktiwiteit belemmer met Botrytis cinerea. Geen inhibisie is getoon teen Penicillium digitatum nie. Hierdie resultate toon die potensiaal van die bio-beheermetode, maar ‘n opskalerings-studie asook ‘n voerlot studie word aanbeveel, veral met die oog op moontlike kommersiële implementering van die strategie. Verdere studies met betrekking tot die aanwendingsmetode van die lipopeptiede moet ook verder ondersoek word, m.a.w. enkel teenoor menigte aanwendigs, aangesien dit die effektiwiteit van die lipopeptiede teen die teikenorganismes sal beïnvloed.

The life-history and infection parameters of the entomopathogenic nematodes Steinernema feltiae (Filipjev)(Nematoda:Rhabditida) and Heterorhahditis megidis (Poinar, Jackson & Klein)(Nematoda:Rhabditida) were examined to provide specific details for the construction of mathematical SI models for biological control of soil insect pests. Laboratory experiments using the Greater Waxmoth, Galleria mellonella as the model host were undertaken to specifically examine the transmission behaviour of infective juvenile nematodes. The proportion of infective juveniles of S. feltiae which infected hosts was dependent on time. Previous studies declared that the proportion of infective juveniles which can infect is static, however, over a period of 5 days most of the infective juveniles infected hosts, demonstrating that the proportion infecting is dynamic. Infection of hosts by both species of nematode was compared using two mathematical representations of the transmission rate. Whereas the most parsimonious form of transmission for H. megidis was the linear Mass Action function, it was evident that, when measured at the individual nematode scale, S. feltiae transmission was non-linear. I postulated that this functional difference is due to the biology of the two species of nematodes. The subsequent effect of including the non-linear response on model predictions were investigated and it was demonstrated that the dynamics of the host nematode interaction became less stable. Spatial models of S. feltiae infection were parameterised from laboratory experiments, and control prediction of these models examined. The horizontal rate of dispersal through sand columns was determined in the presence and absence of hosts. Infective juveniles were found to disperse preferentially towards hosts. The predicted dynamics of pest control using the spatial moqel were highly dependent on the degree of nematode dispersal, host dispersal and the attraction of nematode infective juveniles towards hosts. The overall findings of this thesis have been placed in the context of epidemiological models created elsewhere, and predict that entomopathogenic nematodes may be targeted to specific pest systems with a high degree of success. An understanding of the infection biology of these nematode species is crucial in determining how and when pests may be controlled, and equally importantly, which systems successful control is not predicted.

Decision making regarding the release of biological control agents for invasive species such as lantana, Lantana camara, requires the consideration and evaluation of environmental impact assessment (EIA) reports by a competent authority. Although various biological control agents have been authorised for release into the environment for the control of lantana, the quality of the EIA reports that form the basis for decision making has never been evaluated. The evaluation of the quality of EIA reports on the release of biological control agents by means of an adapted Lee- Colley review package was the focus of this research. The main conclusion was that the quality of the EIA reports on the release of biological control agents for the control of Lantana camara (lantana) was poor by the standards of the review package, the literature reviewed, and the legal requirements. The main deficiencies in the EIA reports related to impact identification, impact evaluation, scoping, mitigation measures and monitoring programmes, while the project descriptions, non-technical summaries and layout and presentation of information in the reports were of good quality. These results correspond to reports in literature that affirms that essential information about impact identification and evaluation and subsequent mitigation and monitoring, the crux of the EIA, is mostly insufficient in EIA reports.
Thesis (M. Environmental Management)--North-West University, Potchefstroom Campus, 2007

[EN] Aphis spiraecola Patch. (Hemiptera: Aphididae) is a key pest of clementines. Biological control of A. spiraecola is still poorly known and efforts were based on the use and conservation of parasitoids but it did not success. With all this said, the aims of this thesis were: i) to disentangle the reasons behind the low parasitism of A. spiraecola; ii) to determine when and how predators can control A. spiraecola populations; and, finally, iii) to evaluate whether a ground cover of Poaceae plants can enhance the biological control of this aphid in clementines by improving the establishment of its predators. In the first objective we sampled four orchards and determine the parasitoid complex and parasitism (and hyper-) rates weekly. Binodoxys angelicae Haliday (Hymenoptera: Braconidae) was the unique primary parasitoid emerged from mummies of A. spiraecola. At least six hymenopteran hyperparasitoid species were identified attacking this primary parasitoid: Syrphophagus aphidivorus (Mayr) (Encyrtidae), Alloxysta sp. (Forster) (Figitidae), Asaphes sp. (Walker) (Pteromalidae), Pachyneuron aphidis (Bouché) (Pteromalidae), Dendrocerus sp. (Ratzeburg) (Megaspilidae) and Phaenoglyphis villosa (Hartig) (Figitidae). We developed a DNA-based approach to untangle the structure of the aphid-parasitoid food web in citrus. This methodology confirmed that all six species hyperparasitized B. angelicae And they dominated this food web and they were abundant from the beginning of the season. Thus, hyperparasitism probably explains the low impact of B. angelicae on A. spiraecola populations. For the second objective we sampled three clementine orchards to determine the effect of aphid predators on A. spiraecola colonies over a three-year period. Life parameters of A. spiraecola colonies varied among the orchards over the three years. The maximum number of aphids and the longevity of A. spiraecola colonies were negatively correlated with the time of first attack by predators. More importantly, the percentage of shoots occupied by A. spiraecola (damages) remained below or close to the intervention threshold when colonies were attacked prior to ~200 degree days (DD) since the beginning of the aphid colonization. These results suggest that: i) the presence of predators at the beginning of the season should be considered to develop new intervention thresholds and ii) biological control programs should promote the early presence of predators in clementine orchards. To promote the early presence of predators in clementine orchards, in the third objective we evaluated ground cover management. This ground cover management may provide alternative preys to natural enemies. The effect of a sown ground cover (based on Poaceae plants) on the biological control of A. spiraecola was evaluated in four orchards with ground cover management compared with four orchards with bare soil management. This sown Poaceae cover coexists with a complex of wild plants that might also affect biological control of A. spiraecola. Finally, we compared the presence of A. spiraecola and its natural enemies in these orchards. While Poaceae plants represented ~66% of the ground cover, the rest of the cover comprised mainly Malva sp. (13%), Oxalis sp. (5%) and Sonchus sp. (2%). Poaceae plants harbored aphids which appeared sooner in the system than citrus aphids. These aphids serve as alternative prey/hosts for natural enemies. By contrast, Malva sp. and Sonchus sp. harbored aphids with potential to become citrus pest. Although these wild plants may act as reservoirs for A. spiraecola as well as other aphid species that can disrupt the biocontrol services of natural enemies, overall, the sown cover was effective in terms of biological control of A. spiraecola in the citrus canopy. It promoted the early presence of predators in citrus canopies. These attacks resulted in satisfactory aphid control, because citrus orchards with ground cover never exceeded the aphid economic threshold.
[ES] Aphis spiraecola Patch. (Hemiptera: Aphididae) es una de las plagas claves en el cultivo de clementinos. Los esfuerzos realizados hasta la fecha se han centrado en el uso y conservación de parasitoides aunque se desconocen las causas de su baja eficacia. Por todo ello, los objetivos de esta tesis han sido i) desentrañar las razones por las que se dan bajos niveles de parasitismo de A. spiraecola ii) determinar cuándo y cómo los depredadores pueden controlar las poblaciones de A. spiraecola y finalmente iii) determinar si una cubierta de poáceas puede mejorar el control biológico de este pulgón en clementinos mediante la mejora en el establecimiento de sus depredadores. En el primer objetivo se muestrearon semanalmente cuatro parcelas y se identificó el complejo de parasitoides y las tasas de parasitismo (e hiperparasitismo). Los porcentajes de parasitismo fueron bajos (~menos del 5%) y Binodoxys angelicae Haliday (Hymenoptera: Braconidae) fue el único parasitoide primario emergido de las momias de A. spiraecola. Se identificaron al menos seis especies de hiperparasitoides atacando este parasitoide primario: Syrphophagus aphidivorus (Mayr) (Encyrtidae), Alloxysta sp. (Forster) (Figitidae), Asaphes sp. (Walker) (Pteromalidae), Pachyneuron aphidis (Bouché) (Pteromalidae), Dendrocerus sp. (Ratzeburg) (Megaspilidae) y Phaenoglyphis villosa (Hartig) (Figitidae). Se desarrolló un método basado en la detección de ADN con el cual se confirmó que todas las especies de hiperparasitoides hiperparasitan B. angelicae. Los hiperparasitoides dominaron esta red trófica y fueron abundantes desde el inicio de la estación. De este modo, el hiperparasitismo probablemente explica el bajo impacto que B. angelicae tiene sobre las poblaciones de A. spiraecola. Para el segundo objetivo se muestrearon tres campos de clementinos donde se determinó el efecto de los depredadores en las colonias de A. spiraecola. Los parámetros de vida de las colonias de A. spiraecola variaron entre los diferentes cultivos los tres años. El máximo número de pulgones y la longevidad de las colonias de A. spiraecola se correlacionaron negativamente con el momento del primer ataque del depredador a la colonia. Cabe destacar que el porcentaje de brotes ocupados por A. spiraecola permaneció por debajo o cerca del umbral de tratamiento cuando las colonias fueron atacadas antes de los 200 grados días (GD) desde el inicio de formación de la colonia. Estos resultados sugieren: i) la presencia de depredadores al inicio de la temporada de pulgón debes ser considerado para el desarrollo de nuevos umbrales de tratamiento y ii) los programas de control biológico deben promover el adelanto de la presencia de depredadores en los campos de clementinos. Para promover la presencia anticipada de depredadores en los campos de clementinos, como tercer objetivo se evaluó el manejo de cubiertas vegetales a base de poáceas. Con este manejo se persigue aportar presas alternativas para los enemigos naturales de A. spiraecola. Para ello, se compararon cuatro campos de cítricos con cubierta vegetal frente a cuatro con suelo desnudo. En los campos con cubierta sembrada apareció además de las poáceas sembradas, un complejo de plantas salvajes que podrían afectar también el control biológico de A. spiraecola. Las poáceas representaron un 66% de la cubierta vegetal. Las poáceas y Oxalis sp. albergaron respectivamente pulgones estenófagos de plantas poáceas y Macrosiphum euphorbiae Thomas (Hemiptera: Aphididae). Estas especies de pulgones aparecieron más pronto en el ecosistema que los pulgones de cítricos y sirvieron como presas/hospederos alternativos para los enemigos naturales. Al contrario, Malva sp. y Sonchus sp. albergaron especies de pulgón que podrían ser potenciales plagas de cítricos. El efecto total de la cubierta sembrada resultó positivo para el control de A. spiraecola. Por lo tanto, las parcelas de cítricos con cubierta vegetal tendieron a no
[CAT] Aphis spiraecola Patch. (Hemiptera: Aphididae) és una de les plagues clau en el cultiu de clementins. Els esforços realitzats fins ara s'han centrat en el us i conservació de parasitoids encara que es desconeix les causes de la seua baixa eficàcia. Tenint en compte estos antecedents, els objectius d'esta tesis foren: i) desentrampar les raons per les quals els parasitoids no són efectius; ii) determinar quan i com els depredadors poden controlar les poblacions d'A. spiraecola; i finalment iii) determinar si una coberta de poàcies pot millorar el control biològic d'este àfid en clementins mitjançant la millora en el establiment del seus depredadors. En el primer objectiu es van mostrejar setmanalment quatre parcel·les i s'identificà el complex de parasitoids i les taxes de parasitisme (i hiperparasitisme). Binodoxys angelicae Haliday (Hymenoptera: Braconidae) va ser l'únic parasitoid primari emergit de les mòmies d'A. spiraecola. Se van identificar al menys sis especies d' hiperparasitoids atacant este parasitoid primari: Syrphophagus aphidivorus (Mayr) (Encyrtidae), Alloxysta sp. (Forster) (Figitidae), Asaphes sp. (Walker) (Pteromalidae), Pachyneuron aphidis (Bouché) (Pteromalidae), Dendrocerus sp. (Ratzeburg) (Megaspilidae) i Phaenoglyphis villosa (Hartig) (Figitidae). Se desenvolupà un mètode basat en la detecció de DNA amb el que es confirmà que totes les especies d'hiperparasitoids hiperparasiten B. angelicae. Els hiperparasitoids dominaren aquesta xarxa tròfica i foren abundants a l'inici de l'estació. Per tant, l'hiperparasitisme podria explicar el baix impacte que B. angelicae té sobre les poblacions d'A. spiraecola. Per al segon objectiu es mostrejaren tres camps de clementins on es determinà l'efecte dels depredadors en les colònies d'A. spiraecola. Els paràmetres de vida de les colònies d'A. spiraecola variaren entre les tres parcel·les els tres anys. El màxim número d'àfids i la longevitat de les colònies d'A. spiraecola es correlacionaren negativament amb el moment del primer atac del depredador a la colònia. Caldria destacar que el percentatge de brots ocupats per A. spiraecola es mantingué per baix o prop del llindar de tractament quan les colònies foren atacades abans dels ~200 graus dia (GD) des de l'inici de formació de la colònia. Estos resultats sugereixen que: i) la presència de depredadors a l'inici de la estació de l'àfid podria ser considerada per al desenvolupament de nous llindars de tractament i ii) els programes de control biològic deurien promoure l'avançament de la presència de depredadors en els camps de clementins. Per promoure la l'avançament de la presència de depredadors en els camps de clementins, com tercer objectiu s'avaluà el maneig de cobertes vegetals basades en poàcies. Amb aquest maneig es persegueix aportar preses alternatives per als enemics naturals d'A. spiraecola. Se compararen quatre camps de clementins amb coberta vegetal front a quatre amb sol nu. En els camps amb coberta sembrada creix, junt a les Poáceas sembrades, un conjunt de plantes salvatges que podrien afectar també el control biològic d'A. spiraecola. S'investigà quines especies de plantes componien la coberta vegetal així com les especies d'àfids que les habitaven. Les poàcies representaren un 66% de la coberta vegetal, sent les plantes salvatges més abundants Malva sp. (13%), Oxalis sp. (5%) i Sonchus sp. (2%). Les poàcies hostejaren àfids estenòfags de poàcies. Estes espècies d'àfid aparegueren més prompte en l'ecosistema que aquells àfids associats a cítrics. Al contrari, Malva sp. i Sonchus sp. hostejaren especies d'àfids que podrien ser potencials plagues de cítrics. Este últim grup pot atenuar l'atac dels enemics naturals a les poblacions d'A. spiraecola que habiten les copes. Encara així, l'efecte total de la coberta sembrada a base de poàcies va resultar positiu per al control d'A. spiraecola. En conseqüència les parcel·les de cítrics am
Gómez Marco, F. (2015). Integrated Pest Management of Aphis spiraecola (Hemiptera: Aphididae) in clementines: enhancing its biological control [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/53732
TESIS

Thesis (MSc)--Stellenbosch University, 2003.
ENGLISH ABSTRACT: Paecilomyces Ii/acinus, ras 251 (geregistreer in terme van wet 36 van 1947 as Suid-Afrika se eerste natuurlike nematisiede en kommersieel beskikbaar as PI Plus) is as biologiese beheer agent getoets by aartappels en in geïntegreerde beheer programme by sitrus en wingerd teen respektiewelik Me/oidogyne species, Ty/enchu/us semipenetrans en verskeie ektoparasitiese nematodes. Die swam toon belofte vir die beheer van hierdie nematodes en het terselfdertyd nie 'n nadelige effek op nie-teiken, voordelige organismes in die grond nie. Veral in kombinasie met chemiese middels, as deel van geïntegreerde programme, kan dit lei tot verminderde gebruik van hoogs toksiese middels en dus meer omgewingsvriendelike landboupraktyke. Biological control of plant parasitic nematodes on potatoes, citrus and grapevine with the fungus, Paecilomyces liIacinus. Paecilomyces liIacinus, race 251 (registered in terms of act 36 of 1947 as South Africa's first natural nematicide, commercially available as PI Plus) was tested as a biological control agent on potatoes and in integrated control programs on citrus and grapevine against Me/oidogyne species, Ty/enchu/us semipenetrans and various ectoparasitic nematodes respectively. The fungus shows promise for the control of these nematodes, without having a harmful effect on non-target, beneficial organisms in the soil. Especially in combination with chemical products, as part of integrated programs, this can lead to less use of highly toxic compounds and thus to more environmentally friendly agricultural practices.
AFRIKAANSE OPSOMMING: Sedert die ontdekking van die swam, Paeci/omyces Ii/acinus (Thom.) Samson as 'n effektiewe eierparasiet van Meloidogyne incognita acrita en Globodera pal/ida (Jatala et al., 1979) het verdere veldproewe in Peru tot die effektiewe beheer van M. incognita en Tylenchulus semipenetrans gelei. Na verskeie suksesse in Peru is die swam onder verskillende klimaat- en grondkondisies in verskeie ander lande beproef. Die sukses behaal in die Filippyne het gelei tot die kommersiële produksie van die swam onder die handelsnaam Biocon. Anders as met chemiese middels vind die werking van biologiese agente stadig oor tyd plaas. Biologiese beheer sal nie chemiese beheer sonder meer kan vervang nie. Dit behoort egter deel te vorm van geïntegreerde nematode bestuur. Inkorporering van die natuurlike organismes, die oordeelkundige gebruik van chemiese nematisiedes, moontlik in kombinasie met die biobeheer agente, weerstand, en ander kulturele praktyke moet ernstig oorweeg word as ons hoop om die steeds groeiende wêreldbevolking te voed (Jatala, 1986). Paecilomyces liIacinus, ras 251, Suid-Afrika se eerste geregistreerde natuurlike nematisiede, kommersieel beskikbaar as PI Plus, is in die Olifantsrivier besproeiingsgebied geëvalueer vir die bestuur van ekonomies belangrike plantparasitiese nematodes by aartappels, sitrus en wingerd. Hierdie gewasse is belangrike bedryfstakke van die streek en is onderhewig aan skade deur nematodes wat die opbrengs nadelig beïnvloed. Chemiese beheer bied slegs 'n korttermyn oplossing vir nematode probleme en skadelike getalle word in 'n kort tyd weer opgebou. Boonop lei dié hoogs toksiese middels tot agteruitgang van die omgewing en sy waterbronne. Die toenemende besorgdheid hieroor en die groot potensiaal van biologiese beheer agente (Jatala, 1986) was die hoofrede vir die werk waaroor hier gerapporteer word.

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

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.

Insecticide resistance and on-going legislation changes on the use of insecticides are likely to reduce their availability for use in agriculture; hence there is an urgent need to develop effective biological controls against these plant pests. Aphids are major insect pests of the agricultural and horticultural sectors. Recent work screened a range of phylloplane-residing bacteria for their ability to kill aphids and used alternative insect targets to determine host specificity. Tests with five other aphid genera indicated the bacteria were also able to kill them. However, the bacteria were generally not effective against non-aphid species including Lepidoptera. I aimed to characterise potential aphid killing pathogens and investigate the killing mechanism. An artificial feeding system with a liquid diet was used to devise a high-throughput screening system to identify pathogenic bacteria against the Green Peach Aphid Myzus persicae (“wild type” insecticide susceptible clones plus insecticide resistant clones). Six bacterial strains were pathogenic to all insecticide susceptible and resistant clones although variation in susceptibility was observed. No single bacterial strain was identified that was consistently more toxic to insecticide resistance clones than susceptible clones, suggesting there was no penalty in resistant clones that makes such clones less fit to bacterial challenge. Pseudomonas poae, which was the most pathogenic to nearly all of aphid clones, was selected for further in-depth analyses. Plant colonisation assays showed that the bacterium could effectively grow and persist on three different plant species. Foliar spray of P. poae did not show any hypersensitive (HR) response and populations (log 5-6) remained stable over three weeks of infestation. Additionally, application of the bacterium to plants before aphid colonisation led to a 68 %, 57 %, 69 % reduction in aphid populations on pre-infested peppers, Arabidopsis and sugar beet plants, respectively. Olfactometer analysis showed that bacterial colonisation of leaves had a deterrent effect on aphids that was not evident for leaves or bacteria alone. Genome analysis of the bacterium revealed three different insecticidal toxins, stress response genes and other pathogenicity-related effector proteins which reflect potential toxicity towards aphids. RNA-Seq was used to examine changes in aphid and bacterial gene expression after 38 h of infection. The altered transcript profiles of the aphid revealed 193 differentially expressed genes and limited gene expression of lysosomal and detoxification genes. 1325 genes were differentially expressed in bacteria, which mainly includes iron acquisition and stress response genes, and putative toxins. Single and combinational deletion, and complementation, of different toxins was conducted. In vitro killing analysis indicated all toxins contributed to aphid killing, with a particularly strong effect seen for one, AprX. Together, these data are being used to understand the molecular basis of aphid mortality to bacterial infection with the aim of utilising them as effective biocontrol agents.

Os agentes de controle biológico têm recebido grande reconhecimento, e o seu uso tem contribuído como um complemento ou substituição de agroquímicos. No entanto, existem poucos estudos sobre o controle biológico da Pinta Preta dos Citros, causada pelo fungo Guignardia citricarpa, o que impede o seu mercado in natura, além de prejudicar a sua exportação pelo uso intensivo de insumos químicos para controlar este patógeno. Os fungos do gênero Trichoderma são agentes de controle utilizados em todo o mundo contra vários fitopatógenos relevantes. Este estudo visou identificar proteínas extracelulares secretadas por T. atroviride T17 e por T.harzianum T1A, eficazes para o controle de G. citricarpa. Por eletroforese bidimensional (2D) foram obtidos perfis de proteínas secretadas por Trichoderma em meio de glicose (controle) e em meio suplementado com micélio desativado de G. citricarpa. As proteínas foram identificadas por LC-MS/MS mostrando que ambas as espécies secretam proteínas diferentes. Foram identificadas 68 proteínas das 178 diferencialmente expressas por T. harzianum, sendo a maioria relacionada aos mecanismos de biocontrole, mesmo no meio controle. Foi verificado que em contato com o patógeno a expressão de proteínas relacionadas com o metabolismo primário diminui. Por outro lado, Trichoderma atroviride mostrou uma maior expressão de proteínas relacionadas com biocontrole na presença de micélio do patógeno. Nesta espécie identificamos 59 proteínas de 116 diferencialmente expressas, principalmente proteínas relacionadas com a degradação da parede celular: α- manosidase, quitinase, mutanase, glicosidase, endoquitinase e, algumas famílias de glicoside hidrolases. Os resultados indicam que estas espécies apresentam um elevado potencial como agentes de controle de G. citricarpa. Os resultados são pioneiros em detalhar a interação de Trichoderrma com G. citricarpa, por meio da análise do secretoma.
Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2016-05-11T14:24:21Z No. of bitstreams: 1 Tese Fernanda Blauth de Lima.pdf: 5347447 bytes, checksum: 314027a9e2af4a7cf6e30be761df9f99 (MD5)
Made available in DSpace on 2016-05-11T14:24:21Z (GMT). No. of bitstreams: 1 Tese Fernanda Blauth de Lima.pdf: 5347447 bytes, checksum: 314027a9e2af4a7cf6e30be761df9f99 (MD5) Previous issue date: 2016-05-11
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES.
Biological control agents (BCA) have received great recognition, and their use has contributed as a complement or replacement of agrochemicals. However, there are few studies on the biological control of Black spot of citrus, caused by the fungus Guignardia citricarpa, which prevents the market in natura, besides damaging their export by the intensive use of chemical inputs to control this pathogen. Trichoderma fungi are the most applied worldwide BCA against various relevant plant pathogens. The purpose of this study was to identify extracellular proteins secreted by T. atroviride T17 and of T. harzianum T1A, which are effective for the control of G. citricarpa. Bidimensional electrophoresis (2D) allowed obtaining the secreted protein profiles of Trichoderma grown in glucose medium (control) and in medium containing inactivated mycelium of G. citricicarpa. From the 178 differentially expressed proteins by T. harzianum, 68 were identified, most of them related to biocontrol mechanisms, even in the control medium. In the presence of the pathogen, the expression of proteins related to the metabolism decreases. On the other hand, Trichoderma atroviride showed higher expression of proteins related to biocontrol pathogen when grown in the presence of pathogen mycelium, when compared to the control medium. From the 116 differentially expressed proteins, 59 were identified, mainly proteins related to fungi cell wall degradation such as α-mannosidase, chitinase, mutanase, glycosidase, endochitinase and, some families of glycoside hydrolases. The results indicate that these species have a high potential as biocontrol agents of G. citricicarpa. These results are pioneers in to detail the Trichoderrma interaction with G. citricicarpa through the secretome analysis.

The alfalfa weevil, Hypera postica (Gyllenhal) (Coleoptera: Curculionidae), remains a serious pest in Virginia despite being under complete biological control in the northeastern U.S. In 1996, a survey of 187 alfalfa fields in Virginia was initiated to determine the current pest status of alfalfa weevil and incidence of natural enemies. Fields located in the Piedmont region of the state had significantly higher alfalfa weevil pressure than those in the Shenandoah Valley and southwestern region. The dominant natural enemy of alfalfa weevil larvae was the parasitoid, Bathyplectes anurus (Thomson). Larval parasitization was lower in the Piedmont compared with the other regions. The entomopathogenic fungus, Zoophthora phytonomi, also killed weevil larvae in all regions. A more intensive ecological investigation of alfalfa weevil population dynamics and mortality was initiated in 1997 to determine why H. postica remains a pest in Virginia, particularly in the Piedmont region. Nine alfalfa weevil populations from three geographic locations were sampled and measured over a period of two years. Results showed that warmer winter temperatures in the Piedmont region contributed to a higher rate of alfalfa weevil oviposition compared with the Shenandoah Valley and southwestern mountains. Parasitization of adult weevils by Microctonus aethiopoides (Loan) was low in all regions of Virginia relative to rates reported in the northeastern U.S. This also contributes to relatively high alfalfa weevil fecundity in Virginia because a greater percentage of adults reproduce. Phenological asynchrony between M. aethiopoides and the alfalfa weevil may explain the low parasitism. The 1st generation of M. aethiopoides adults did not emerge until most of the overwintering alfalfa weevil adults had reproduced and died. A majority of alfalfa weevil eggs were laid before January at all locations in Virginia. Approximately half of the egg population survived to contribute to spring larval infestations. In the northern U.S., very few alfalfa weevil eggs survive the winter, and larval populations result primarily from spring-laid eggs. Mortality of larvae was high in Virginia and comparable to that reported in other states. Bathyplectes anurus was well-synchronized with alfalfa weevil and killed a high percentage of larvae.
Ph. D.

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.

During last century rising populations and changed market preferences have led to large structuralchanges in agriculture in the developed world. Most conventional cultivation methods involve theproduction of few monoculture crops within a homogenous landscape, where pesticides and inorganicfertilizers are used to increase yields. The cereal aphid Rhopalosiphum padi (L.) is a pest in cerealsresulting in large economic losses for farmers throughout Europe, and are currently removed withchemical pest control. However, the pesticides also affect existing ecosystem services like potentiallypest-controlling insects as well as the surrounding environment negatively. According to previousstudies higher landscape diversity leads to a higher diversity of natural enemies to pest insects, but nostudy has investigated if the pest control is enhanced around non-crop remnants such as midfield islets(MI). This thesis investigates (i) if MI will reduce the number of aphids in crop field, (ii) if larger MIwill be more effective than smaller MI by having higher aphid predation rate, and (iii) if larger MI willprovide a more effective reduction of pest insects over a longer distance from edge. To assess thepotential for pest control, aphids were glued on papers and placed at ground level in crop fields andaround MI in nine fields. The collected data was processed statistically, and the result demonstratesthat MI play an important role by providing habitats for natural enemies to pests. The increased size ofMI have a positive effect of number of aphids consumed around MI, however the distance from edgewas not significance. This study highlights the value of MI in crop fields as an existing ecosystemservice in biological pest control. The naturally occurring predators are able to reduce R.padi andthereby minimize the need for insecticide applications.
Ökat befolkningstryck och större efterfrågan av jordbruksprodukter på marknaden har det senasteseklet lett till stora förändringar i västvärldens jordbrukslandskap. Odlingslandskapet är idaghomogent och monokulturellt där besprutningsmedel används för att ge större avkastning iproduktionen. Skadeinsekten havrebladlus (Rhopalosiphum padi) angriper vårgrödor vilket leder tillstora ekonomiska förluster för bönder runtom i Europa. Pesticider används för att reducera angreppenmen påverkar även miljön negativt samt existerande ekosystemtjänster, såsom naturliga fiender sombiologiskt kan begränsa skadeinsekters angrepp. Tidigare studier visar att heterogena landskap har enhög biologisk mångfald, men ingen studie har undersökt åkerholmars (ÅH) eventuella potential förbiologisk bekämpning av skadeinsekter i säd. Denna studie undersöker därför (i) om ÅH leder tillökad reducering av havrebladlöss i säd, (ii) om större ÅH är mer effektiva än mindre i frågan ompestreducering, samt (iii) om större ÅH begränsar lusangrepp längre ut i fält från ÅH:s kant än mindreholmar. Studiens hypoteser testades genom experiment utförda i nio fält med vårgrödor där lössklistrades på etiketter som placerades ut i omgivande fält. Dessa samlades in efter tjugo fyra timmarför att studera antal löss konsumerade i relation till ÅH. Det insamlade materialet är bearbetatstatistiskt och resultatet visar att ÅH spelar en betydande ekologisk roll för naturliga fiender tillskadeinsekten havrebladlus. Fler löss konsumerades runt större ÅH än de med mindre yta eller i de fältutan ÅH. Däremot har avståndet från ÅH en mindre betydelse, vilket indikerar att pestreduceringen ärmest effektiv nära stora ÅH. Denna studie synliggör värdet av existerande ÅH som inhysernyttoinsekter i jordbrukslandskapet, samt att heterogena landskap har högre potential för en mernaturlig skadedjursbekämpning. Naturliga fiender är därmed en existerande ekosystemtjänst som kanbegränsa bladlusangreppen (R.padi), som i sin tur ger möjligheten att reducera användandet avbekämpningsmedel, vilket för oss närmare miljömålet ”en giftfri miljö”.

Thesis (doctoral)--Swedish University of Agricultural Sciences, 2002.
Thesis documentation sheet inserted. Appendix reprints five published papers and manuscripts, three co-authored with others. Includes bibliographical references. Also available electronically via World Wide Web in PDF format; online version lacks appendix.