O uso de pesticidas constitui um importante aspecto na agricultura moderna, com inquestionável beneficio na produção agrícola. Porém a contaminação dos alimentos por pesticidas constitui um sério risco a saúde do consumidor. A determinação de resíduo de pesticidas em alimentos envolve procedimentos laboriosos, com elevado tempo de análise e várias etapas de pré-concentração. Neste trabalho, procedimentos alternativos de extração e pré-concentração para analise multiresíduo de pesticida em água, frutas e tubérculos foram desenvolvidos. A eletroforese capilar em seu modo MEKC em condições de alto e baixo fluxo eletrosmótico foi empregado para a otimização da separação de diferentes classes de pesticidas (triazinas, organofosforado, carbendamidazóis, feniluréia e carbamatos). A composição do eletrólito de separação otimizada para condições de alto EOF foi: 10 mmol L-1 de tetraborato de sódio (pH 9,3),50 mmol L-1 de SDS e 5% etanol e 5% propanol, enquanto que para condições de baixo EOF foi: 10 mmol L-1 de tampão fosfato (pH 2,5), 25 mmol L-1 de SDS e 10% metanol. Estratégias de pré-concetração on-line conhecida como sweeping (SW) e stacking nos modos de migração reversa das micelas (SRMM) e migração reversa com um plug de água (SWR) bem como as suas versões modificadas foram avaliadas, obtendo fatores de pré-concetração de variaram de 2,6 a 19 para o SW, 2,9 a 15 para o SRW e de 5,5 a 15 para o método SRMM modificado. Varias metodologias de extração envolvendo extração em fase sólida (SPE) e extração líquido-líquido (LLE) foram testada. A estratégia de extração por cloud point foi aplicada a uma amostra de abacaxi. O procedimento denominado dispersão da matriz em fase sólida (sigla inglesa MSPD), que minimiza o uso de solventes orgânicos e é de fácil implementação foi aplicado a amostras de cenoura. A combinação do SPE off-line e das estratégias de pré-concentração anteriormente mencionadas permitiram a determinação de alguns pesticidas na concentração de 0,1 &#181;g L-1 em amostras de água potável. O método de extração e clean-up MSPD seguida da análise de MEKC em alto EOF foi otimizado e algumas figuras de mérito foram estabelecidas baseados em protocolos de validação para análise de pesticidas (IAEA-FAO). Boa linearidade (r > 0,99) foi obtido para todos os pesticidas estudados, exceto para linuron e dimetoato. A precisão do método foi estimada através de testes de recuperação. Dois níveis de fortificação foram utilizados para a avaliação, foram obtidos recuperações de 51 a 89 % para o nível mais baixo e 67 a 100% para o maior nível. Foi obtida uma boa precisão intraensaio (CV < 15%). O método otimizado foi aplicado para análise multiresíduo de cenouras. Uma amostra adquirida no comércio local foi quantificada encontrando-se 0,88 mg kg-1 de simazina, 0,13 mg kg-1 de atrazina e 0,08 mg kg-1 de propazina. / The use of pesticides constitutes an important aspect of modem agriculture, with unquestionable impact on crop production. However, food contamination by pesticide residues is a serious risk for the consumer. The determination of pesticide residues in food usually involves laborious procedures, with time consuming sample clean up and preconcentration steps prior to the analysis. In this work, alternative methodologies for extraction, pre-concentration and analysis of pesticide multi-residue in water, fruits and tubers were developed. Capillary electrophoresis in its micellar mode (micellar electrokinetic Chromatography, MEKC) under low and high electroosmotic flow (EOF) conditions was used for the separation of pesticides from different chemical classes (triazines, organophosphorous, carbendamidazols, phenilurea and carbamates). Optimized electrolyte compositions were: high EOF - 10 mmol L-1 tetraborate (pH 9.3), 50 mmol L-1 SDS, 5 % ethanol and 5 % propanol; low EOF - 10 mmol L-1 phosphate buffer (pH 2.5), 25 mmol L-1 SDS and 10 % methanol. On-line preconcentration strategies for MEKC known as sweeping (SW) and stacking with reverse migrating micelles with (SRW) and without (SRMM) a plug of water prior to the sample plug as well as modified versions of SRW and RMM were evaluated and contrasted in terms of signal enhancement factor (peak height ratios) Signal enhancement factors for SW varied from 2,6 to 19 for SRW from 2,9 to 15, whereas for modified-SRMM from 5,5 to 15. Among the extraction methodologies, several procedures involving solid-phase extraction (SPE) and liquid-liquid extraction (LLE) were tested. It is worth mentioning a strategy based upon cloud point extraction, which was applied to pineapple samples and a procedure denominated matrix solid-phase dispersion (MSPD), which combines low cost, saves in solvents and easy implementation, applied to carrots. The combination of off-line SPE and the above mentioned on-line preconcentration strategies allowed the determination of selected pesticides in the 0.1 &#181;/L level (drinking water sample). A complete methodology involving MSPD for extraction and sample clean-up followed by MEKC in high EOF was optimized and a few figures of merit were established based on method validation protocols for pesticide analysis (IAEA-FAO). Good linearity (r>0.99) was obtained for all pesticides under investigation, except for linuron and dimetoate. The method accuracy was estimated by recovery tests. Two level standard spiking were conducted with recoveries of 51 to 89 % for the lowest level and 67 to 100 % for the highest level. Acceptable intra-day precision was obtained (CV < 15 %). The optimized method was applied to the analysis of multi-residue pesticides in carrots. In a sample acquired in a local grocery store an unusual amount of triazines was found: simazine (O,88 mg/kg), atrazine (0,13 mg/kg) and propazine (0,088mg/kg).

Le concept de « service écosystémique », en plein essor depuis la publication du Millenium Ecosystem Assessment en 2005, a permis de souligner l'importance des performances non-marchandes des écosystèmes. En arboriculture, assurer une bonne productivité tout en préservant les ressources naturelles et la santé humaine est aujourd’hui un défi majeur, qui peut être analysé sous l’angle des services écosystémiques. Quels sont ces services au sein d’un verger de pommiers ? Comment les analyser ? Quels sont les liens - conflits et synergies - entre services écosystémiques multiples et comment les systèmes de culture modifient-ils les profils de services multiples? Ce travail de thèse vise à répondre à ces questions avec une démarche novatrice combinant mesures expérimentales, modélisation et analyse statistique. Suite à une analyse bibliographique des services écosystémiques pouvant être délivrés dans des vergers, cinq services ont été sélectionnés. Il s’agit de la production de fruits, de la disponibilité de l’azote dans le sol, de la régulation du climat reposant sur la prévention de la dénitrification de l’azote et sur la séquestration du carbone, de la maintenance et de la régulation du cycle de l’eau y compris de sa qualité, et de la régulation des bioagresseurs. Conjointement à ces services nous avons considéré les nuisances environnementales dues aux pesticides. Pour chaque service nous avons identifié les fonctions écosystémiques sous-jacentes ainsi que les pratiques agricoles et les conditions pédoclimatiques ayant un impact sur ces fonctions. Les services et fonctions ont été décrits par un ou plusieurs indicateurs et quantifiés à l’aide de modèles dans les cas suivants : (i) neuf systèmes de culture réels sur deux dispositifs expérimentaux dans le sud-est de la France et (ii) 150 systèmes de culture fictifs conçus à partir de la combinaison de cinq leviers de pratiques et de leurs modalités, dans des conditions pédoclimatiques identiques. Les deux modèles utilisés ont été STICS, un modèle générique de simulation du fonctionnement du sol et des cultures sous l’influence des pratiques, qui a nécessité une paramétrisation et une évaluation sur pommier à partir de mesures expérimentales, et IPSIM, une plateforme de modélisation simulant les effets des pratiques et des conditions du milieu sur les dégâts aux cultures causés par les bioagresseurs. IPSIM a été paramétré sur pommier, sur la base d’une importante étude bibliographique et à dires d’experts. Les simulations des modèles ont fait l’objet d’analyses statistiques simples dans le cas des systèmes de culture réels, et d’analyses multivariées à deux tableaux (analyse en composantes principales avec variables instrumentales) pour les systèmes de culture fictifs. Pour les neufs systèmes de culture réels, 14 liens importants entre services écosystémiques ont été identifiés, notamment des conflits comme celui entre la prévention de la dénitrification ou de la lixiviation de l’azote et la disponibilité de l’azote dans le sol à court terme, et des synergies, comme celle entre l’humidité du sol ou la séquestration du carbone et la disponibilité de l’azote dans le sol à court terme. Ces liens entre services sont expliqués par les fonctions écosystémiques sous-jacentes. La comparaison de profils de services entre systèmes a mis en évidence l’impact de pratiques agricoles sur certains services. Ainsi, sur un même site, une forte densité de plantation augmente la production de fruits et la séquestration du carbone. La fertilisation exclusivement sous une forme organique diminue la production de fruits via un stress azoté mais diminue également la lixiviation. Par ailleurs, les profils de services sont fortement influencés par les caractéristiques pédoclimatiques de chaque site. Ces résultats confortent la nécessité d’une prise en compte explicite de l’interdépendance ‘pratiques x conditions du milieu’ pour analyser les services. / The concept of « ecosystem service », which has been used increasingly since the publication of the Millennium Ecosystem Assessment in 2005, has highlighted the importance of ecosystem’s non-marketed performances. In orchards, ensuring high productivity while preserving natural resources and human health has become a real challenge that could be analyzed with the concept of ecosystem service. Which ecosystem services are delivered in an apple orchard? How to analyze them? What are the relationships - conflicts or synergies – among multiple ecosystem services and how do cropping systems change multiple ecosystem service profiles? This PhD work aims at answering those questions with an innovative approach combining experimental measures, modeling and statistical analysis.Based on a literature review of ecosystem services in orchards, five services were selected: fruit production, nitrogen availability in soil, climate regulation based on the prevention of nitrogen denitrification and on carbon sequestration, maintenance and regulation of water cycle, including water quality, and pest control. We also considered the environmental disturbances caused by the use of pesticides. For each service, we identified the underlying ecosystem functions as well as the agricultural practices and soil and climate conditions affecting these functions. Services and functions were described by one or multiple indicators and quantified using models in the case of (i) nine existing cropping systems on two experimental sites in southeastern France differing in terms of soil and climate conditions, and (ii) 150 virtual cropping systems designed out of the combination of five major agricultural practice levers and their modalities, in identical soil and climate conditions. The two models used were STICS, a generic soil-crop simulation model under the influence of practices which required a parameterization and an evaluation on apple orchards based on experimental measures, and IPSIM, a generic modeling framework simulating the impacts of agricultural practices and local conditions on crop injuries caused by pests. IPSIM was parameterized on apple orchards, based on an important literature review and expert opinions. Model simulations were analyzed with simple statistics in the case of the nine existing cropping systems and with two-table multivariate analyses (principal component analysis with instrumental variables) for virtual cropping systems.Concerning the existing cropping systems, 14 important relationships were identified among ecosystem services, especially conflicts, like the one between nitrogen denitrification or leaching prevention and soil nitrogen availability on the short term, and synergies such as the one between soil humidity or carbon sequestration and nitrogen availability on the short term. These relationships are explained by the underlying ecosystem functions. Comparing service profiles among cropping systems highlighted the impacts of agricultural practices on some services. That way, on a same site, a high planting density increases fruit production and carbon sequestration. An exclusively organic fertilization decreases fruit production through nitrogen stress but also nitrogen leaching in drained water. Furthermore, service profiles are strongly influenced by the soil and climate conditions of each site. These results strengthen the need to explicitly consider the ‘agricultural practices x soil and climate conditions’ interdependence in order to analyze ecosystem services. The results obtained with the virtual cropping systems simulations confirmed those of the existing ones and gave precision on the impacts of fertilization, irrigation and pest control for codling moth, rosy apple aphid and apple scab on ecosystem functions and services.

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2018-05-28T15:31:34Z No. of bitstreams: 1 CAMILA MARIA FORMIGA LEITE - DISSERTAÇÃO PPGSA PROFISSIONAL 2016..pdf: 1330991 bytes, checksum: 3763a8f8c7326718476ac867c8c63cb6 (MD5) / Made available in DSpace on 2018-05-28T15:31:34Z (GMT). No. of bitstreams: 1 CAMILA MARIA FORMIGA LEITE - DISSERTAÇÃO PPGSA PROFISSIONAL 2016..pdf: 1330991 bytes, checksum: 3763a8f8c7326718476ac867c8c63cb6 (MD5) Previous issue date: 2016-12-15 / O feijão-caupi representa uma fonte de alimento estratégica pelos altos teores de proteínas, carboidratos e de sais minerais e adaptação às condições ambientais adversas. Existem pragas capazes de atacar grãos/feijão armazenados, e a mais importante dessas no Brasil é o Callosobruchus maculatus (Coleoptera: Bruchidae), destacando-se por atacar sementes perfeitas, devido ao seu potencial depreciativo, trazendo má qualidade aos grãos e consumindo as reservas de nutrientes do embrião. O método de controle mais utilizado contra pragas de armazenamento é o uso de produtos químicos de distintas classes toxicológicas. Embora esses produtos tenham bastante eficiência, seu uso intensificado pode acarretar em problemas, com isso a utilização das plantas com atividade inseticida tornou-se tão importante, sendo geralmente aplicados na forma de pós, extratos ou óleos. Diante do exposto, este trabalho teve como objetivo avaliar o efeito repelente e inseticida de ervas utilizadas na cultura popular como medicinais no controle de Callosobruchus maculatus em grãos de feijão-caupi (Vigna unguiculata L.) em condições de laboratório, analisando a dose letal (DL50) e tempo letal (TL50) desses carunchos em relação ao pó dessas erva medicinais (Melissa officinalis (erva cidreira), Peumus boldus (boldo-do-chile), Maytenus ilicifolia (espinheira-santa), Mentha (hortelã). O experimento foi conduzido no laboratório de Entomologia da Universidade Federal de Campina Grande (UFCG), Campus Pombal, Paraíba, Brasil. Os grãos de feijão-caupi foram tratados com os pós nas concentrações de 5,0 g e em doses diferenciadas de 0,25, 0,50, 0,75 e 1,0 % (massa do pó/massa de grãos) e realizados testes de preferência e sobrevivência contra C. maculatus. No que se refere à repelência Erva cidreira, Boldo-do-chile e a Hortelã os insetos exibiram propriedades repelentes, melhor especificando os insetos não tiveram preferência pelas mesmas. Já com a Espinheira Santa, os insetos mostraram-se atraentes/preferentes. No que se referem à sobrevivência todas as espécies causaram mortalidade, em determinado período de tempo. / The Cowpea represents a strategic food source for the high levels of proteins, carbohydrates and mineral salts and adaptation to adverse environmental conditions. There are plagues capable of attacking grains/beans stored, and the most important of these in Brazil is the Callosobruchus maculatus (Coleoptera: Bruchidae), stand out to attack perfect seeds due to its derogatory potential, bringing bad quality grains and consuming nutrient reserves of the embryo. The most widely used control method against pests is the use of chemical products of different toxicological classes. Although these products have a lot of efficiency, its use can cause problems if it’s intensified, then the use of plants with insecticidal activity became so important, being generally applied in the form of powders, extracts or oils. On the exposed, this work aimed to evaluate the effect of insecticide and repellent herbs used in popular culture as the medical control of Callosobruchus maculatus in Cowpea beans, (Vigna unguiculata L.) under laboratory conditions, analyzing the lethal dose (Ld50) and lethal time (TL50) of these weevils in relation to those medicinal herb powder (Melissa officinalis (balm), Peumus boldus (boldo), Maytenus ilicifolia (espinheira-santa), Mentha (Mint). The experiment was conducted in the laboratory of Entomology, Federal University of Campina Grande (UFCG), Campus Pombal, Paraíba, Brazil. Cowpea beans were treated with powders at concentrations of 5.0 grams and different doses of 0.25, 0.50, 0.75 and 1.0% (mass/mass of powder grains) and preferably tests and survival against c. maculatus. As regards the Lemongrass repellency, Boldo and Mint insects exhibited repellent properties, better specifying the insects did not have the same preference. The Maytenus, the bugs proved attractive/preferences. In referring to the survival every species have caused mortality at any given period of time.

Aerial control using 1080 (sodium monofluoroacetate) baits is widely used in New Zealand for the control of introduced brushtail possums (Trichosurus vulpecula), with the aim of protecting national conservation and agricultural values from these damaging pests. This thesis integrates research, completed over 25 years, that was motivated by growing recognition in the 1970s of the extent of possum impacts and the need to improve the effectiveness and efficiency of the control operation. Field research assessed the palatability of three types of cereal-based pellet baits and carrot baits in different regions, habitat types and seasons. Palatability was assessed by the consumption of the different bait types presented independently of each other on 15-30 plots, with rotation of bait types at plots on successive nights to provide equal exposure to each bait type. There was regional variation in possums' bait preferences, possibly reflecting genotypic differences, whereas seasonal variation was less evident. Carrot bait was preferred or equally preferred to cereal bait in 14 out of 20 field trials. The proportion of possums eating baits was then investigated by, firstly, developing a technique for tracing bait acceptance using rhodamine B, a UV-fluorescent dye. In four field trials, more than 95% of possums accepted three types of dye-marked bait, eliminating bait refusal as a major reason for low kills in winter control operations. In a fifth trial, conducted in summer, only 68% of possums accepted bait suggesting that seasonal availability of favoured foods may influence bait acceptance. Since possums must encounter baits before deciding whether to eat them, field studies were undertaken to assess the coverage achieved in normal aerial baiting operations. Large gaps, up to 400 m in width, were often found between baiting swaths; these could allow some possums to survive. A controlled field experiment, using acceptance of rhodamine-dyed bait as a measure of effectiveness, showed that bait distribution was least accurate where flight paths were not marked. Where gaps of 100 m between flight paths were deliberately created, bait acceptance was slower and less than where coverage was complete. Sowing baits at 3 kg/ha was as effective as at 10 kg/ha, indicating the potential for substantially reducing operational costs by using machinery capable of faultlessly distributing baits at low rates. Navigational guidance systems were evaluated and found to improve the accuracy of bait distribution. During 1993-1997, when a lower sowing rate of 5 kg/ha was adopted operationally by regional managers, control effectiveness was unchanged but annual savings of around $9 million accrued. Because of the lack of suitable sowing machinery, a bucket was developed to permit faultless distribution of baits at lower rates, demonstrating the possibility of yet further cost-savings. The possibility of seasonal food availability affecting bait acceptance was investigated in three different forest habitats. Dyed baits were aerially distributed on 100 ha at each site in each season over two years. In each trial, fat-based condition indices of possums were calculated and the abundance of possum-preferred plant foods described. Bait acceptance was consistently high (85-100%) in the 24 trials, and was not influenced by either condition or availability of preferred foods. It seems likely that seasonal variation in operational effectiveness is caused by either the availability of sharply seasonal, scarce foods that possums may feed on intensively for brief periods, or by warmer temperatures that render 1080 less effective. The influence of 1080 on acceptance of (rhodamine-dyed) baits was investigated in a field trial. Examination of possums for dye-marking showed that 25% of possums refused to eat either a lethal quantity of bait or any bait at all, compared with 98% of possums eating non-toxic bait. This indicated that 1080 is aversive to possums, which is a potential major reason for their surviving control operations. Pen trials were therefore conducted to further examine the problem and to seek solutions. Toxic carrot baits were rejected by 27.5% of possums, equally by smell and taste aversion, whereas toxic cereal pellets were rejected by 34%, mainly by taste aversion. Orange and cinnamon were shown to be among the most preferred of 42 flavours tested and, when applied to toxic baits, 1080 was effectively masked. Bait refusal was reduced to ≤7%, the same as that recorded for possums presented with flavoured non-toxic baits. For long-term control of possum populations, aerial 1080 baiting can be used sequentially with other poisoning methods. However, the compatibility of these methods is dependent on the likelihood of possums developing bait shyness if sublethally dosed. Studies were therefore conducted to characterise and compare the four main toxicants used (1080, cyanide, cholecalciferol and brodifacoum) for induction and mitigation of bait shyness. Shyness was induced in approximately 80% of possums sublethally dosed with cyanide, 60% with 1080, 20% with cholecalciferol, and 0% with brodifacoum. Cyanide and 1080 shyness were found to persist in many possums for at least 12 and 24 months, respectively. Use of alternative bait types, and of baits containing an alternative slow-acting toxin (brodifacoum) were shown to be effective ways of overcoming shyness. This, and other related research, is reviewed to provide operational specifications that maximise the likelihood that all targeted possums will (i) encounter bait, (ii) eat it, and (iii) die. The likely future use of aerial 1080 baiting is described and the technological, economic, environmental and social constraints on its sustainability are discussed. Finally, the uptake of the research by possum managers is considered, and areas identified in the thesis where information is incomplete are summarised as prioritised topics for further research.

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”.

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

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

Crop rotation may be used to prevent the continuous selection of particular weed species adapted to one crop type. This might be useful for weed management, economy in herbicide applications and promoting biodiversity. Common simple crop sequences might be diversified by introducing perennial forage crops. Impacts of such perennial crops on weeds were studied with four approaches : 1) Large-scale weed surveys in 632 fields in western France showed that weed species composition differed most strongly between perennial alfalfa crops and annual crops. Comparisons of fields before, during and after perennial alfalfa suggested that community composition varies in a cyclic way during such crop rotations. Several weed species problematic in annual crops were suppressed during and after perennial crops, but the appearance of other species led to equal or even higher plant diversities. 2) A 3-year field experiment with contrasting crop management options allowed an investigation of the underlying mechanisms for this: The absence of soil tillage reduced weed emergence but increased the survival of established plants. The permanent vegetation cover and frequent hay cuttings reduced weed growth, plant survival and seed production. 3) Greenhouse experiments testing the regrowth ability of individual plants after cutting showed strong differences between species and functional groups. An two-factorial experiment suggested that the negative impacts of cutting and competition on weed growth were mainly additive. 4) Special measurements of weed seed predation in the field experiment showed positive correlations with vegetation cover, indicating that this ecosystem service may be particularly fostered by perennial crops. Consistent preferences of seed predators for certain weed species indicates that seed predation may be another cause of the observed weed community shifts.

Parthenium hysterophorus (Asteraceae), classified as an emerging weed in South Africa, has become abundant throughout large parts of southern and eastern Africa. In South Africa it has invaded areas in KwaZulu-Natal, Mpumalanga, the North West Province and Limpopo. A biological control programme against parthenium weed was launched in South Africa in 2003, based on the success achieved in Australia. Zygogramma bicolorata, a leaf-feeding beetle native to Mexico, was imported into South Africa via Central Queensland, Australia where it was released in the 1980s. This thesis examines aspects of the thermal physiology of Z. bicolorata which, in conjunction with its native and exotic geographical distribution, was used to predict the potential distribution of the agent in South Africa, in relation to climate. To determine Z. bicolorata’s physiological capability, several physiological parameters were examined for mechanistic modelling purposes. These parameters included the beetle’s lethal thermal limits, critical thermal limits, lethal humidities (Chapter 2) and developmental rate at constant temperatures (Chapter 3). In Chapter 4, these physiological parameters were entered into the dynamic modelling program CLIMEX (CLIMEX programme ver. 2, CSIRO Entomology ©) and a map of the areas that are acceptable for the establishment of Z. bicolorata was produced. The CLIMEX model predicted that most of South Africa is favourable for the establishment of the beetle, except in the west of the country and in the north of Lesotho, extending into South Africa. All areas in which parthenium currently occurs were predicted to be very favourable for Z. bicolorata establishment and proliferation. Optimal release sites aimed at initial establishment were earmarked at three areas in the northeastern part of South Africa (Jozini, Ndumu Game Reserve and along the road from Swaziland to Mozambique). It is concluded that Z. bicolorata is climatically suited to South Africa, increasing the likelihood that populations will establish and proliferate when released. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.