Thesis (MSc (Genetics))--University of Stellenbosch, 2010. / Includes bibliography. / Title page: Dept. of Genetics, Faculty of Science / ENGLISH ABSTRACT: Next-generation sequencing technologies are increasingly used in metagenomic studies, largely due to the high sequence data throughput capacity and unbiased approach in determining the genetic composition of an unknown environmental sample. This study investigated the applicability of the Illumina next-generation sequencing platform for metagenomic sequencing of grapevine viruses to provide the first complete viral profile, or virome, of a diseased vineyard. Leaf material was harvested from 44 randomly selected vines in a leafroll-diseased vineyard in South Africa. Sample material was pooled and double-stranded RNA extracted. The dsRNA was sequenced as a paired-end sequencing run using the Illumina sequencing-by-synthesis technique, and more than 19 million sequence reads, equivalent to approximately 837 megabases of metagenomic sequence data, were obtained. Of these data, approximately 400 megabases could be assembled into 449 scaffolds, using the de novo assembler Velvet. These scaffolds were subjected to BLAST searches against the NCBI databases and top hit scores were used for virus identification. Based on the BLAST results, suitable sequences were selected from the NCBI database and used as reference sequence in MAQ mapping assemblies. The bioinformatic analyses allowed for the determination of the virus species present, the most prominent variants, and the relative abundance of each. Four known grapevine viral pathogens were identified. Grapevine leafroll-associated virus 3, representing 59% of the analyzed short read sequence data, was identified as the most prominent virus species. Three variants of this virus were detected: GP18 was the most abundant, followed by a minor Cl766/NY1 variant and a potential novel grapevine leafroll-associated ampelovirus. A single Grapevine rupestris stem pitting ]associated virus variant, similar to SG1, and a Grapevine virus A variant, a member of molecular group III, were identified. This study is also the first to report the presence of Grapevine virus E (GVE) in South African vineyards. Grapevine virus E was further genetically characterized and the genome sequence of GVE isolate SA94 determined. The GVE SA94 genome sequence, 7568 nucleotides in length, is the first complete genome sequence for the virus species. The genome organization of GVE SA94 is typical of vitiviruses, but in contrast to other RNA viruses, the AlkB domain is located within the helicase domain in open reading frame 1 (ORF 1). Grapevine virus E SA94 shares nearly 100% nucleotide identity with the Japanese TvP15 isolate and GVE 3404, a de novo scaffold generated from the metagenomic sequence data. Bioinformatic analysis of metagenomic sequence data further revealed the presence of three fungus-infecting viral families, Chrysoviridae, Totiviridae and the unclassified dsRNA virus, Fusarium graminearum dsRNA mycovirus 4. A virus from the family Chrysoviridae, similar to Penicillium chrysogenum virus, was the second most abundant virus detected. We demonstrated the successful application of a short read sequencing technology, such as the Illumina platform, for viral profiling of an infected vineyard. To our knowledge this is the first application of the Illumina technology for this purpose. / AFRIKAANSE OPSOMMING: Volgende-generasie tegnologie om basis volgordes van nukleiensure te bepaal, word al meer gebruik in metagenomiese studies. Dit is veral weens die hoe data-omset kapasiteit en onbevooroordeelde aanslag in die bepaling van die genetiese samestelling van onbekende omgewingsmonsters. Hierdie studie het die aanwending van die Illumina volgende-generasie volgorde-bepalingsplatform in 'n metagenomiese studie van wingerdvirusse, ondersoek. Dit het ten doel gehad om die eerste volledige virus profiel, of viroom, van 'n geinfekteerde wingerd saam te stel. Blaarmateriaal is verkry vanaf 44 lukraak-gekose wingerdstokke in 'n rolblad-geinfekteerde wingerd in Suid-Afrika. Monster materiaal is saamgevoeg en dubbelstring-RNS geekstraheer. Die dubbelstring-RNS is onderwerp aan gepaarde-ent volgorde-bepaling deur gebruik te maak van die Illumina volgorde-bepaling-deur-sintese tegniek. Meer as 19 miljoen volgorde reekse, ekwivalent aan ongeveer 837 megabasisse volgorde data, is verkry. Van hierdie data kon ongeveer 400 megabasisse saamgevoeg word in 449 konstrukte ("scaffolds"), deur gebruik te maak van die de novo samesteller Velvet. Hierdie konstrukte is onderwerp aan BLAST soektogte teen die NCBI databasisse en die hoogste trefslag-telling is gebruik vir virus identifikasie. Op grond van die "BLAST" resultate is geskikte volgordes geselekteer vanaf die NCBI databasis en gebruik as verwysingvolgordes in MAQ kartering-analises. Met die bioinfomatika analises kon die virus spesies teenwoordig, asook die mees prominente variante en relatiewe voorkoms van elk, bepaal word. Vier bekende virus wingerdpatogene is geidentifiseer. Grapevine leafroll-associated virus 3, verteenwoordig deur 59% van die geanaliseerde kort-reeks volgorde data, is identifiseer as die mees prominente virus spesie. Drie variante van die virus is in die wingerdmonster opgespoor: GP18 kom die mees algemeen voor, gevolg deur 'n CL-766/NY1 variant en 'n potensiele nuwe wingerd rolblad-geassosieerde ampelovirus. 'n Enkele Grapevine rupestris stem pitting-associated virus variant, soortgelyk aan SG1, en 'n Grapevine virus A variant, 'n lid van molekulere groep III, is geidentifiseer. Hierdie studie is ook die eerste om die teenwoordigheid van Grapevine virus E (GVE) in Suid-Afrikaanse wingerde te rapporteer. Grapevine virus E is verder geneties gekarakteriseer en die genoomvolgorde van GVE isolaat SA94 is bepaal. Die GVE SA94 genoomvolgorde, 7568 nukleotiede lank, is die eerste volledige genoomvolgorde vir hierdie virus spesie. Die genoomorganisasie is tipies van vitivirusse, maar in kontras met ander RNA virusse is die AlkB domein binne-in die helikase domein van oopleesraam 1 (ORF 1) geleë. Grapevine virus E SA94 deel byna 100% nukleotied identiteit met die Japannese TvP15 isolaat en GVE 3404, 'n de novo konstruk gegenereer vanaf die metagenomiese volgorde data. Bioinformatika analises van die metagenomiese volgorde data het verder die teenwoordigheid van drie swam-infekterende virus families, die Chrysoviridae, Totiviridae en ongeklassifiseerde dubbelstring-RNS virus, Fusarium graminearum dsRNA mycovirus 4, aangetoon. 'n Virus van die Chrysoviridae familie, soortgelyk aan Penicillium chrysogenum virus, het die tweede meeste voorgekom in die wingerd monster. Hierdie studie demonstreer die suksesvolle toepassing van 'n kort reeks volgorde-bepalingstegnologie soos die Illumina platform, vir die opstel van 'n virusprofiel van 'n geinfekteerde wingerd. Sover ons kennis strek is hierdie die eerste aanwending van die Illumina tegnologie vir hierdie doel.

Thesis (MScAgric)--University of Stellenbosch, 2001. / ENGLISH ABSTRACT: Apple replant disease (ARD) is one of the major impediments to the establishment of an economically viable apple orchard on sites previously planted to apple. In spite of extensive research on ARD, the etiology remains to be fully elucidated. A possible biological origin of ARD etiology in South Africa was investigated by the dilution of replant field soil with sterilised soil. Commercial orchards with ARD were selected for use in pot trials and disease severity evaluated after three months, by measuring shoot length, dry mass of plants as well as root discolouration. Although diluting replant soil to 25 and 50% (v/v) significantly reduced the effects of ARD, symptoms were only absent in 0% replant soil. It was clear that seedlings planted in any mixture containing replant soil, even only 25% replant soil, consistently exhibited symptoms of stunted growth and root discolouration similar to those seedlings grown in 100% replant soil. This indicates that ARD in South Africa is primarily of a biological nature. As an initial step in formulating sustainable disease control alternatives to replace methyl bromide, pot trials were conducted to assess the impact of compost treatments as well as biological control products on ARD. Compost as well as sterilised and unsterilised compost teas (compost extract) significantly increased seedling growth even under optimum nutrient conditions when compared to the control, suggesting that they negate the effects of ARD. Results also indicated that applying high concentrations of compost does not necessarily provide additional growth benefits compared to lower concentrations. Results with biocontrol formulations were less favourable. Only one of the biocontrol formulations, a combination of Bacillus spp. (Biostart®) improved growth significantly compared to the control. There was, however, some inconsistency with results for the different trials conducted using this product. Fungal as well as nematode populations associated with ARD soils were characterised to the generic level to get a clearer understanding of the etiology of ARD in South Africa. Pythium and Cylindrocarpon spp. were consistently isolated from all six replant soils in all trials that formed part of this study, indicating that these fungi may have a role in ARD etiology in South Africa. Nematodes implicated in ARD development were inconsistently associated with ARD soils used in these studies. This suggests that nematodes do not have a primary causal role in ARD etiology in South Africa. Field trials were conducted in commercial orchards to assess the impact of organic amendments and promising biological control products, as indicated by the pot trials, on ARO severity under field conditions. These biological soil amendments were also compared with the standard chemical control methods for ARO, methyl bromide and chloropicrin. In all three trials established, compost and mulch as well as manure and mulch, consistently increased growth to the same extent as the standard chemical treatments and by combining these chemical treatments with organic amendments a significant, additional growth increase could be attained. Biocontrol formulations evaluated in field studies gave variable results. Biostart® improved growth when applied on its own, but not in combination with the chemical Herbifume (metham-sodium). Inoculating soil with effective microorganisms (EM), consisting primarily of photosynthetic bacteria, had no significant effect on growth. Results from this study indicate that application of organic amendments could possibly substitute for soil fumigation in replanted apple orchards. However, compost quality standards need to be implemented and because few types of compost are universally effective, different types of composts should be compared in specific soil environments before recommendations can be made. Oue to variable results with biocontrol products, ARO management with these biological soil amendments cannot be guaranteed at this stage and further studies are recommended. / AFRIKAANSE OPSOMMING: EVALUERING VAN DIE SPESIFIEKE APPELHERVESTIGING-PROBLEEM IN BOORDE IN DIE WES-KAAP Appelhervestiging-siekte (AHS) skep 'n groot probleem in die vestiging van jong appelbome op grond waar daar reeds voorheen appels verbou is. Ten spyte van omvangryke navorsing is die oorsaak van die probleem nog hoofsaaklik onbekend. 'n Moontlike biologiese oorsaakleer is in Suid-Afrika ondersoek deur die hervestigings-effek te probeer verminder deur die vermenging van hervestigingsgrond met gesteriliseerde grond. Kommersiële boorde met 'n appelhervestigingsprobleem is geselekteer en gebruik in potproewe. Die ernstigheidsgraad van die siekte is na drie maande se groei geevalueer deur lootlengte, droë massa en wortelverkleuring te meet. Alhoewel verdunning van die hervestigingsgrond tot 50 en 25% (vlv) die effek van AHS op groei betekenisvol verminder het, kon die skadelike effek van die veroorsakende faktor slegs uitgeskakel word deur saailinge in 100% gesteriliseerde grond te plant. Dit was duidelik dat saailinge wat in enige grondmengsel geplant is waarin hervestigingsgrond voorgekom het, selfs al was dit net 25%, konsekwent simptome van vertraagde groei en wortelverkleuring getoon het. Dit is 'n aanduiding dat AHS in Suid-Afrika hoofsaaklik biologies van aard is. Potproewe is uitgevoer as 'n eerste stap in die formulering van volhoubare siektebeheer-strategieë, om die impak van kompos-behandelings en biologiese beheer produkte op AHS te ondersoek. Kompos sowel as gesteriliseerde en ongesteriliseerde kompos-tee (kompos-water) het, selfs onder optimale voedingsomstandighede, die groei van saailinge betekenisvol verbeter. Dit dui aan dat hierdie behandelings die effek van AHS kan teenwerk. Resultate het ook daarop gedui dat hoër kompos konsentrasies nie noodwendig enige addisionele voordele vir groei inhou in vergelyking met laer konsentrasies nie. Resultate met biologiese beheer produkte was minder gunstig. Slegs een van die produkte wat geëvalueer is, 'n kombinasie van Bacillus spp. (Biostart®), het groei betekenisvol verbeter in vergelyking met die kontrole. Resultate was egter inkonsekwent vir die verskillende proewe waarin hierdie produk gebruik is. Swampopulasies sowel as aalwurmpopulasies wat met hervestigingsgrond geassosieer word, is geïdentifiseer tot op generiese vlak om vas te stel waardeur AHS in Suid-Afrika veroorsaak word. Pythium en Cylindrocarpon spp. is konsekwent van al ses hervestigingsgronde geïsoleer wat daarop dui dat hierdie twee swamgenera 'n beduidende rol in AHS ontwikkeling in Suid-Afrika mag hê. Aalwurms wat aangedui is in die literatuur om 'n moontlike rol in AHS te hê, was slegs in enkele gevalle geassosieer met hervestigingsgronde waarvan in hierdie studie gebruik gemaak is. Die gevolg-trekking is dus gemaak dat aalwurms nie 'n betekenisvolle rol speel as hoof-veroorsakende organisme onder Suid-Afrikaanse toestande nie. Veldproewe is uitgevoer in kommersiële appelboorde om vas te stel wat die effek van organiese materiaal, asook belowende biologiese beheermiddels, soos aangedui deur potproewe, op AHS onder veldtoestande is. Die biologiese grondtoedienings is ook vergelyk met die standaard chemiese beheermiddels (metielbromied en chloorpikrien). In al drie proewe wat gevestig is, het kompos met 'n deklaag, sowel as kraalmis met 'n deklaag, groei betekenisvol verbeter tot dieselfde mate as chemiese middels. Daar kon ook 'n beduidende, addisionele groeitoename gemeet word in gevalle waar chemiese middels met organiese materiaal gekombineer is. Resultate met biologiese beheer formulasies wat onder veldtoestande geëvalueer is, het gevarieer. Biostart® het groei verbeter wanneer dit alleen toegedien is, maar in kombinasie met die chemiese middel Herbifume (metham-sodium) het dit geen effek gehad nie. Die inokulering van grond met 'n oplossing van effektiewe mikroorganismes (EM) wat hoofsaaklik uit fotosinterende baterieë bestaan, het ook geen betekenisvolle effek op groei gehad nie. Die gevolgtrekking is gemaak dat toediening van organiese materiaal moonlik as plaasvervanger vir metielbromied-beroking kan dien in die beheer van AHS. Die nodige komposkwaliteit-standaarde moet egter eers geïmplimenteer word. Omdat feitlik geen kompos universeel effektief kan wees nie, is dit ook nodig dat verskillende tipes kompos met mekaar vergelyk moet word in spesifieke grondtoestande voordat verdere aanbevelings gemaak kan word. As gevolg van variërende resultate met biologiese beheer produkte kan AHS beheer met hierdie middels nie gewaarborg word op hierdie stadium nie en verdere studies word aanbeveel.

Although a considerable amount of work has been done on the fauna of the soil of temperate forests and pastures, very little is known about that of tropical and sub-tropical soils. Most of the work on tropical soils concerns the fauna of tropical forests ... . The present investigation represents an attempt to discover the effect of an imported perennial tropical and sub-tropical plant, the Pineapple, on the soil fauna of a semi-arid region (rainfall 15-25 inches). Intro., p. 1-2

The potato tuber moth, Phthorimaea operculella (Zeller), is a major pest of potato crops worldwide causing significant damage to both field and stored tubers. The current control method in South Africa involves chemical insecticides, however, there is growing concern on the health and environmental risks of their use. The development of novel biopesticide based control methods may offer a potential solution for the future of insecticides. In this study a baculovirus was successfully isolated from a laboratory population of P. operculella. Transmission electron micrographs revealed granulovirus-like particles. DNA was extracted from recovered occlusion bodies and used for the PCR amplification of the lef-8, lef-9, granulin and egt genes. Sequence data was obtained and submitted to BLAST identifying the virus as a South African isolate of Phthorimaea operculella granulovirus (PhopGV-SA). Phylogenetic analysis of the lef-8, lef-9 and granulin amino acid sequences grouped the South African isolate with PhopGV-1346. Comparison of egt sequence data identified PhopGV-SA as a type II egt gene. A phylogenetic analysis of egt amino acid sequences grouped all type II genes, including PhopGV-SA, into a separate clade from types I, III, IV and V. These findings suggest that type II may represent the prototype structure for this gene with the evolution of types I, III and IV a result of large internal deletion events and subsequent divergence. PhopGV-SA was also shown to be genetically more similar to South American isolates (i.e. PhopGV-CHI or PhopGV-INDO) than it is to other African isolates, suggesting that the South African isolate originated from South America. Restriction endonuclease profiles of PhopGV-SA were similar to those of PhopGV-1346 and PhopGV-JLZ9f for the enzymes BamHI, HindIII, NruI and NdeI. A preliminary full genome sequence for PhopGV-SA was determined and compared to PhopGV-136 with some gene variation observed (i.e. odv-e66 and vp91/p95). The biological activity of PhopGV-SA against P. operculella neonate larvae was evaluated with an estimated LC₅₀ of 1.87×10⁸ OBs.ml⁻¹ being determined. This study therefore reports the characterisation of a novel South African PhopGV isolate which could potentially be developed into a biopesticide for the control of P. operculella.

Two ant species, Pheidole megacephala (Fabricius) and Anoplolepis custodiens (Smith) reach pest status in citrus orchards through precipitating outbreaks of homopterous pests. However, predacious ants, including these two ant species, play an important role in pest suppression in agroecosystems and are therefore beneficial to these systems. If A. custodiens and P. megacephala are important natural control agents in citrus, using ant bands to break the mutualism between the ants and the Homoptera as a method of ant control is preferable to poisoning. Ant communities were sampled by pitfall trapping in three experimental subunits of 2-, 4-, 15- and 30-year-old citrus orchards, in the Grahamstown area of the Eastern Cape. In one subunit in each orchard, populations of P. megacephala and A. custodiens were suppressed by poison applications. In a second subunit, trees were banded with trunk barriers so that ants were prevented from foraging in the trees and a third subunit served as the untreated control. Bait pupae of bollworm, false codling moth and fruit fly were planted in bait trays in all of the subunits to investigate predation on these citrus pests in the relative absence of predacious ants and where they were excluded from the trees. Pheidole megacephala dominated exclusively in all of the plots. Community composition did not change dramatically with increasing age of the trees, but species diversity and species abundance did. Rank-abundance curves showed that community diversity was highest in the 2-year-old plots and lowest in the 30-year-old plots. The Simpson and Shannon-Wiener diversity indices and their evenness measures indicated that diversity and equitability were highest in the poisoned subunits and lowest in the banded subunits. Principle component analysis revealed that the poisoned subunits were similar and distinct in species composition, that there was significant monthly variation in species composition and that community stability increases with an increase in orchard age. The presence of P. megacephala was significantly negatively correlated (rs = -0.293; p < 0.001) with pest pupal survival. Pupal survival was significantly higher for bollworm (p < 0.001), FCM (p < 0.001) and fruit fly (p < 0.001) in the poisoned subunits, than in the banded and control subunits. There was a general trend for survivorship to increase with an increase in the age of the trees. A significant difference (p < 0.001) was found between the months in which the trials were carried out. Pupal survival was significantly lower (p < 0.001) for FCM than for bollworm and fruit fly. In citrus orchards, ant communities are organised by ecological processes and interactions and are influenced by methods of ant control. Ant bands are preferable to poisoning as a method of ant control, so that beneficial species are left on the ground to prey on pests that pupate in the soil. Maintaining high ant species diversity in citrus orchards would be beneficial as predation on the pupae was more effective where ant species diversity was higher.

MSc (Zoology) / Department of Zoology / See the attached abstract below

PhD (Agric) (Plant Production) / Department of Plant Production / Brassicas are important vegetable crops grown for home consumption and market gardening in eastern and southern Africa. However, productivity is affected by aphids, through both direct feeding and disease transmission. Botanical insecticides have potential to control the aphids, but so far few plants have been evaluated for use on brassicas. This study was conducted to evaluate the effectiveness of Solanum panduriforme to control aphids on brassicas. Botanical extracts from three parts of S. panduriforme were assessed for their insecticidal effects on the cabbage aphid, Brevicoryne brassicae. The extracts from leaf powder (LP), ripe berry powder (BP), fresh ripe berries (RB) and fresh unripe berries (UB) were extracted with four solvents; water, ethanol, hexane and diethyl ether, using homogenisation, maceration and solvent-assisted / sequential extraction methods. The effectiveness of the extracts was determined by laboratory bioassays as well as by plant assays in the screen house and under field conditions. The experiments were replicated three or four times depending on the assays and the design used was completely randomized design (CRD). The immature (LP and UB) plant parts were generally more effective than the mature (BP and RB) plant parts, with mortalities ranging from 100 % down to 40 % respectively depending on assays. Ethanol extracts were more effective than aqueous extracts (LP 96% and 63%; BP 96% and 64%; RB 100% and 64%; UB 100% and 90%). The dried crude extracts from hexane were more effective than di-ethyl ether extracts. The group chemical analysis indicated presence of alkaloids in the berries (BP, RB and UB), which were absent in the leaves (LP). Phenolic compounds and flavonoids were present in all the extracts (LP, BP, RB, and UB). Saponins were present in the fresh parts (RB and UB). The results show how the locally available S. panduriforme plants can be used as an aphicide to control aphids on brassicas. Farmers can directly prepare an easy and cheap botanical / NRF

Please read the abstract in the section 07chapter7. / Dissertation (MSc (Genetics))--University of Pretoria, 2002. / Genetics / unrestricted

Thesis (MScAgric)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: The grain chinch bug, Macchiademus diplopterus, is an endemic pest of cultivated grain crops and wild grasses in the south-western Cape region of South Africa. In early summer when host plants dry out, adult grain chinch bugs aggregate in large numbers in shelter sites in surrounding areas and enter into aestivation. These shelter sites sometimes include the stalk or calyx ends of fruit, and shelter-seeking bugs can also contaminate export fruit cartons, consequently posing a phytosanitary/quarantine risk to importing countries. Presently, there are no feasible pre- or post-harvest control measures to manage this quarantine risk. The aggregating behaviour of grain chinch bugs suggests the involvement of pheromones. Therefore, investigating the chemical ecology of grain chinch bugs for potential use in control measures is the focus of the first research chapter of this study. Gas chromatography-mass spectrometry (GC-MS) was used to identify headspace volatiles collected from aggregating bugs. Olfactometer bioassays were conducted to assess the attractiveness of each gender to separate sexes, individual compounds and a mixture of the compounds as a formulated lure. The lure was tested in field trapping trials with delta and bucket traps. In the bioassays with the live insects the response of each gender to live females was greater than the responses of each gender to live males, suggesting that females may disseminate the pheromones more efficiently than males. The following eight volatile compounds were indentified from the GC-MS analysis: hexanal, (E)-2-hexenal, (E)-2-hexenol, (E)-2-hexenyl acetate, (E)-2-octenal, (E)-2-octenol, (E)-2-octenyl acetate and tridecane. In the bioassays with individual compounds, three of these eight compounds, hexanal, (E)-2-hexenal, and tridecane, elicited attraction of both females and males. The formulated lure was attractive to both males and females in the laboratory bioassay, but this attraction was not evident in the field. In the field, there was only one occasion when a significantly higher number of bugs were caught in baited traps compared to unbaited traps. Trap catches were very low compared to the actual level of infestation in the field which was evident from corrugated cardboard bands tied around tree trunks which contained many sheltering bugs. The low trap catches seen in the field were partly due to competition between the synthetic pheromone lure and the natural pheromones emitted by aggregating live insects. Also, the characteristic shelter-seeking behaviour of grain chinch bugs influenced trap catches, as more bugs were found in places that provide shelter, like cardboard bands and walls of the delta traps. This behavior of aestivating bugs could be used to the advantage of trapping bugs by integrating sheltering sites into traps in future trials. Also, the lure needs to be improved for optimum efficiency in the field. The second research chapter also addresses the quarantine risk posed by grain chinch bugs, by investigating the thermal biology of bugs to ultimately facilitate the development of effective post-harvest treatments. Critical thermal minimum and maximum temperatures (CTmin and CTmax) of both active and aestivating bugs were subjected to critical thermal limits analysis. The CTmin and CTmax of aestivating bugs were not affected by gender (p > 0.05). There was a decrease in CTmin from the active period into aestivation for both males (2.8°C to 1.0°C (± 0.1)) and females (2.1°C to 0.6°C (± 0.1)). Also, for CTmax there was an increase in tolerance from the active period into the aestivation period for both males (49.9°C to 51.0°C (± 0.1)) and females (49.9°C to 51.5°C (± 0.1)). To determine the plasticity of grain chinch bug thermal tolerance, aestivating bugs at 27 weeks into aestivation, were acclimated at different temperatures and photoperiods [18°C (10L:14D) and 26°C (16L:8D)] for a period of seven days. Both low (18°C) and high (26°C) acclimation temperatures and photoperiods increased CTmin of aestivating grain chinch bugs at 14 weeks from 0.8°C to -1.2°C and -0.1°C (± 0.1) respectively. However, CTmax was not altered by acclimation temperatures (p > 0.82). Field temperatures at collection sites were recorded to compare to grain chinch bugs thermal tolerance levels exhibited in the laboratory. These results, as well as the effects of acclimation treatments on the CTmin of bugs, have implications for post-harvest treatments, and understanding the quarantine risk posed to importing countries. The information generated from this study can be used to further advance the development of both effective pre-harvest and post-harvest control measures to reduce grain chinch bug quarantine risk. / AFRIKAANSE OPSOMMING: Die graanstinkluis, Macchiademus diplopterus, is 'n endemiese plaag van aangeplante graangewasse en wilde grasse in die Suidwes Kaap-provinsie van Suid-Afrika. In die vroeë somer wanneer gasheerplante uitdroog, soek groot getalle volwasse graanstinkluise skuiling in die omliggende gebiede en gaan in ʼn somerrusperiode. Hierdie skuilplekke sluit soms die stam of kelk eindes van vrugte in en graanstinkluise kan ook uitvoer-vrugte kartonne kontamineer. Gevolglik word lande wat vrugte uit Suid-Afrika invoer, aan die fitosanitêre kwarantynrisiko van stinkluisbesmetting blootgestel. Tans is daar nie haalbare voor- of na-oes beheermaatreëls om hierdie kwarantyn risiko te bestuur nie. Die aggregasiegedrag van graanstinkluise dui op die betrokkenheid van ʼn feromoon. ‘n Ondersoek van die chemiese ekologie van die graanstinkluis vir moontlike gebruik in beheermaatreëls is die fokus van die eerste gedeelte van hierdie studie. Gaschromatografie-massaspektrometrie (GC-MS) is gebruik om die vlugtige organiese verbindings in die bodamp van die saamgetrosde stinkluise te identifiseer. Olfaktometriese biotoetse is uitgevoer om die aantreklikheid van die insekte vir die teenoorgestelde geslag te bepaal, asook van die individuele verbindings en 'n mengsel van die verbindings as 'n geformuleerde lokmiddel in lokvalle. Die lokmiddel is getoets in veldproewe met deltatipe en emmertipe lokvalle. In die olfaktometriese biotoetse met die lewende insekte is die reaksie van beide geslagte teenoor lewende wyfies groter as die reaksie van die geslagte teenoor mannetjies, wat daarop dui dat wyfies die feromoon meer doeltreffend as mannetjies versprei. Die volgende agt verbindings is geïdentifiseer met behulp van GC-MS-analise: heksanaal, (E)-2-heksenaal, (E)-2-heksenol, (E)-2-heksenielasetaat, (E)-2-oktenaal, (E)-2-oktenol, (E)-2-oktenielasetaat en tridekaan. In die biotoetse met individuele verbindings het drie van die agt verbindings, hexanal, (E)-2-hexenal, en tridecane, lokaktiwiteit vir beide geslagte getoon. Die geformuleerde lokmiddel was aantreklik vir beide geslagte in laboratorium toetse, maar soortgelyke lok is nie in die veld gevind nie, waar daar net een keer 'n aansienlike groter getal graanstinkluise met lokmiddel gevang is in vergelyking met lokvalle sonder lokmiddel. Die getal graanstinkluise in lokvalle was baie laag in vergelyking met die werklike vlak van besmetting in die veld, wat duidelik geblyk het uit die getalle graanstinkluise wat skuiling gesoek het in die geriffelde karton bande wat om boomstamme vasgemaak was. Die lae lokvalvangste in die veld was deels te wyte aan die kompetisie tussen sintetiese feromoon en die natuurlike feromoon van saamgetrosde insekte. Die kenmerkende aggregasiegedrag van graanstinkluise het lokvalvangste beïnvloed, aangesien meer stinkluise gevind is in plekke wat skuiling bied, soos die kartonbande en die binnekant van die delta-lokvalle. Hierdie skuilings van graanstinkluise kan in toekomstige proewe uitgebuit word deur vir meer skuilplek in lokvalle voorsiening te maak. Die formulering en die aanbieding van die lokmiddle moet ook verbeter word vir 'n optimale doeltreffendheid in die veld. In die tweede hoofstuk word die kwarantynrisiko van die graanstinkluis aangespreek deur die ondersoek van die termiese biologie van stinkluise om uiteindelik die ontwikkeling van doeltreffende na-oes behandelings te fasiliteer. Kritiese termiese minimum en maksimum temperature (CTmin en CTmax) van beide aktiewe en rustende graanstinkluise is bepaal deur analise van die kritiese termiese beperkings van die insek. Die CTmin en CTmax van rustende graanstinkluise is nie geraak deur geslag nie (p > 0.05). Daar was 'n afname in CTmin van die aktiewe tydperk tot in rus, vir beide manlike (2.8°C tot 1.0°C (± 0.1)) en vroulike insekte (2.1°C tot 0.6°C (± 0.1)). Ook vir die CTmax was daar 'n verbetering in toleransie vanaf die aktiewe tydperk tot in die rusperiode vir beide manlike (49.9°C tot 51.0°C (± 0.1)) en vroulike insekte (49.9°C tot 51.5°C (± 0.1)). Om die aanpasbaarheid van die termiese toleransie van die graanstinkluis te bepaal, is graanstinkluise 27 weke na aanvang van die rusperiode geakklimatiseer by verskillende temperature en fotoperiodes [18°C (10L: 14D) en 26°C (16L: 8D)] vir 'n tydperk van sewe dae. Beide lae (18°C) en hoë (26°C) akklimatiseringstemperature en fotoperiodes het onderskeidelik die CTmin van rustende graanstinkluise op 14 weke verhoog van 0.8°C tot -1.2°C en -0.1°C (± 0.1). Daar is egter geen effek op CTmax deur akklimasie temperature nie (p > 0.82). Veldtemperature is ook bepaal om te vergelyk met graanstinkluis termiese toleransie vlakke wat in die laboratorium bepaal is. Hierdie resultate, sowel as die gevolge van die akklimasie behandelings op die CTmin van graanstinkluise, het implikasies vir na-oes behandelings, en begrip van die kwarantyngevaar wat dit inhou vir vrugte-invoerlande. Die inligting wat uit hierdie studie voortvloei, kan gebruik word om die ontwikkeling van beide effektiewe voor-oes en na-oes beheermaatreëls te bevorder en om die kwarantynrisiko wat graanstinkluise inhou, te verminder.

This study assessed the mycological and aflatoxin contamination of peanuts collected from Kinshasa, DRC and Pretoria, South Africa. Forty peanut samples were collected randomly at informal markets in the two cities and analysed for mycoflora and aflatoxins (B1, B2, G1 and G2) using standard methods. The results indicated that 95% and 100% of peanut samples collected from Kinshasa and Pretoria, respectively were contaminated with aflatoxigenic fungi with Kinshasa’s samples being the most contaminated (up to 49, 000 CFU/g). Seventy percent (70 %) of Kinshasa-samples and 35% of Pretoria-samples exceeded the maximum allowable limit of aflatoxin B1 set by JECFA (5 ppb). Statistical evidence showed a significant positive correlation between mycoflora and aflatoxin level for Kinshasa-samples (r = 0.4743, p < 0.005) while Pretoria-samples showed no correlation. The study reveals that high level of contamination in Kinshasa-samples could be due to the tropical nature of the climate and poor storage conditions as compared to Pretoria which is sub-tropical and sanitary regulations are enforced. / Life & Consumer Sciences / M. Sc. (Life Sciences)