Faculty of Science (ETDs)

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Subject: search.filters.subject.SDG-15: Life on land

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    Assessing the effectiveness of wetlands in the Krugersdorp Game Reserve in attenuating pollution from mines on the West Rand, South Africa
    (University of the Witwatersrand, Johannesburg, 2023) Sawuka, Noluthando Thulisile; Evans, Mary; Masindi Khuliso
    In South Africa, 48% of the country’s wetlands are critically endangered because of anthropogenic activities. Wetlands are an important part of the landscape and play a critical role including but not limited to improving water quality, habitat provision, and water storage. This research aimed to assess the effectiveness of wetland systemsin attenuating pollution from water discharged from abandoned gold mines in the Krugersdorp Game Reserve (KGR), West Rand. Eight (8) water samples were collected in the study site. Physico-chemical parameters were measured in situ, and chemical parameters were measured in the lab. The measured physico–chemical parameters from the majority of the sampled wetlands exceeded at least one of the stipulated water quality legislations, which included the General Authorization Limit Section 21f and h, 2013; Unit for TWQGR; Mine Health and Safety Act; and WUL wastewater in terms of the recorded pH, total dissolved solids, and salinity variables. Overall, a decreasing trend in pH level was observed from wetlands sampled upstream of the KGR to wetlands sampled downstream of the KGR, with the highest recorded pH level (Alkalinity: 8.9) obtained from the sampled wetland that was closest to the adjacent mining site upstream of theKGR whilst the lowest recorded pH level (Acidity: 3.9) obtained from a wetland sampling point that was further from the adjoining mine and downstream in the KGR. A weak and positive correlation (r=0.040) was obtained between the measured total dissolved solids and pH levels from the sampled wetlands, indicating minimal spatial variability. However, a strong positive correlation (r=0.999, Correlation is significant at the 0.01 level) was obtained between the measured total dissolved solids and salinity from the sampled wetlands. At least one of the limits stipulated by the water quality legislation was exceeded in terms of the analysed inorganic constituents from the sampled wetlands. The dominant ions recorded in the wetlands in increasing order are F, K, Cl, Mg, Na, Ca, and SO4. Mn and Si were the dominant metal concentrations recorded in most wetlands, with the former also showing exceedances when compared to the stipulated water quality guidelines. The recorded data from the measured physico–chemical parameters and analysed chemical variables indicated poor water quality in wetlands sampled downstream of the KGR and upstream of the KGR. Stringent measures in water quality monitoring need to be implemented to mitigate the environmental impacts associated with wastewater discharge into the receiving environment.
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    The Equidae from Gladysvale, a Hominin locality in the Cradle of Humankind, South Africa
    (University of the Witwatersrand, Johannesburg, 2023-10) Van der Merwe, Daniel Jean; Badenhorst, Shaw
    The later Pleistocene compared to modern census data indicated that Equidae are represented lower than comparably sized bovids (BOV III & IV). The reason for this may be due to numerous factors that have been previously investigated such as, landscape usage, carnivore and prey relationships and accumulation factors. However, a largely unexplored factor that may also have played a vital role in their lower representation may have been the difference and effectiveness of predator avoidance behaviours utilized by Equidae. The Equidae remains from the external and internal deposits of Gladysvale (730 to 580 Kya and 257 to 195 Kya) in the Cradle of Humankind, may provide insight into the predatory avoidance behaviours of two different Equidae species, Equus capensis and Equus quagga. This study aimed to create and renew an understanding of the role of these animals within larger faunal communities in the Cradle of Humankind. Equidae fossil material mostly from Gladysvale along with supporting material from Sterkfontein, Kromdraai and Coopers were examined, measured and photographed, in an attempt to broaden the understanding of Equidae relations within the Cradle of Humankind. Furthermore, to also elaborate on the potential differences between Equus capensis and Equus quagga. Past research has usually assigned Equus capensis to the larger version of Equus quagga during the later Pleistocene, with other studies based on DNA analysis suggesting the two species have very little intraspecific diversity. However as seen in this study it would appear that there is a notable difference in size between the two Equidae species, but also a difference in overall representation within the Cradle of Humankind. This could imply that not only are the two species different in size but that they also employed different predatory avoidance behaviours.
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    Interfacial engineering of NbSe2 and TaSe2 to enhance their electrocatalytic activity for hydrogen production
    (University of the Witwatersrand, Johannesburg, 2023-07) Kolokoto, Tshwarela; Moloto, Nosipho
    There has been a need to replace fossil fuels, develop sustainable energy systems, and alleviate the negative environmental effects. These effects can be alleviated by developing efficient processes such as water-splitting, which can produce hydrogen gas in an environmentally friendly manner and, in turn, use it as a clean fuel. However, this process requires an effective electrocatalyst comparable to Pt and cost-effective. Herein, we demonstrate that the electrocatalytic activity of NbSe2 and TaSe2 can be improved by metal inclusion using interfacial engineering for the hydrogen evolution reaction (HER). The readily synthesised NbSe2 was decorated with 20% wt. Ni, 20% wt. Pt, 10% wt. Pt / 10% wt. Ni using two synthetic methods, namely the ex-situ and in-situ methods. The ex-situ samples had higher HER activities than the in-situ samples. Pt/PtO2-NbSe2 (derived from Pt decorated NbSe2 using the ex-situ method) showed a significantly enhanced HER activity compared to bare NbSe2. The Pt/PtO2-NbSe2 nanomaterial had the lowest overpotential, favourable kinetics and durability in an alkaline solution of 0.1 M KOH. The trend was as follows: Pt/PtO2-NbSe2 (Pt-decorated ex situ) > PtO-NbSe2 (Pt-decorated in-situ) > PtO/NiO-NbSe2 (Pt/Ni-decorated) > Ni/NiO-NbSe2 (Ni-decorated ex-situ) > Ni0.5Se/Ni(OH)2-NbSe2 (Ni-decorated in-situ) > NbSe2. In addition, NbSe2 was further decorated with 20% wt. Co using both the ex-situ and in-situ synthetic methods, and 10% wt. Pt / 10% wt. Co using the in-situ method. The ex-situ sample resulted in a higher HER activity compared to the in-situ samples. In particular, Co/Co3O4-NbSe2 nanomaterial (Co-decorated ex-situ) had the lowest overpotential, favourable kinetics and durability in an alkaline solution of 0.1 M KOH. The resultant trend was as follows: Co/Co3O4-NbSe2 (Co-decorated ex-situ) < Co3O4/CoSe2/PtO/PtO2-NbSe2 (Pt/Co-decorated in-situ) < Co3O4/CoSe2-NbSe2 (Co-decorated in-situ) < NbSe2. Consequently, the ex-situ method was the optimum synthetic method for forming NbSe2-based nanomaterials. TaSe2-based nanomaterials were formed similarly. TaSe2-based hybrids were formed by decorating TaSe2 with 20% wt. Ni, Co and Pt using the ex-situ method. The hybrid nanomaterials resulted in higher HER activities compared to pristine TaSe2 (i.e. Pt/PtO/PtO2-TaSe2 (Pt-decorated) > Ni/Ni(OH)2-TaSe2 (Ni-decorated) > Co/Co3O4-TaSe2 (Co-decorated) > TaSe2). Pt/PtO/PtO2-TaSe2 hybrid, in particular, resulted in the lowest overpotential under alkaline solutions (0.1 M KOH). Generally observed, was NbSe2-based electrocatalysts were better than TaSe2-based catalysts. In addition, the Pt-decorated ex-situ NbSe2 and Pt-decorated TaSe2 electrocatalysts were better than the model Pt/C catalyst, with the prior being the best overall. This is attributed to the basal sites of the NbSe2 and TaSe2. The ex-situ method was better than the in-situ method and this was due to the presence of metallic particles and the minimization of oxidation compared to the latter.
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    Home range establishment of translocated African elephants (Loxodonta africana) in a savanna environment
    (University of the Witwatersrand, Johannesburg, 2023-09) Ngorima, Patmore; Parrini, Francesca; Boyers, Melinda; Mariotti, Elena
    Data from nine female African elephants translocated to Hurungwe Safari Area that had been collared was used to estimate home range patterns. Monthly home ranges for each elephant were estimated using the 50% and 95% Kernel Density Estimates (KDEs) method for a period of 20 months following translocation. At the population level, a linear regression model was used to test for the effect of months from release on the home range patterns. The findings revealed that the 95% home range of the elephants increased over time, while the 50% home range decreased since their initial release. I went further to compare the individual differences using the generalised additive model. The results showed some individual differences in patterns of exploration, with some displaying more linear home range patterns than others. Over and above, a considerably positive and relevant relationship between time and variation in the extent of the home range was observed.
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    The function of “plant biltong” collected by bush Karoo rats (Otomys unisulcatus)
    (University of the Witwatersrand, Johannesburg, 2024) Sanweni Siyabong Patrick
    Food storing is a commonly observed strategy in many rodents living in environments with seasonally fluctuating food availability. Stored food allows small mammals to survive through food restricted periods and has been extensively studied in the northern hemisphere, characterised by cold winters. It could also be adaptive in semi-deserts of the southern hemisphere, such as the Succulent Karoo, which experience winter rainfall and hot dry summers. Here I want to understand the function of food plants carried back by bush Karoo rats (Otomys unisulcatus) to the stick lodges they build in the Succulent Karoo. Some of the food plants carried back to their lodges dry out, forming what I refer to as “plant biltong” or ‘plantong.’ To date, the function of the plantong remains unknown. Plantong might function as food in the dry season, or it might just be left over from food eaten at the lodge, or it can aid in construction of the lodges. I studied which plant species were collected as plantong, and whether there were seasonal differences in collection and consumption of plantong. I observed 15 lodges over 12 months and recorded the fate of plant pieces carried back to each lodge by bush Karoo rats. I tested experimentally whether they do eat the plantong. Plant species that were collected and dried out to become plantong were mostly annual shrubs (Zygophyllum retrofractum, Lycium cinereum, Salsola zeyheri, Helichrysum), grass species (Schismus) and perennials species (Atriplex, Drosanthemum spec, Galenia sarcophylla, Hermannia, Mesembryanthemum, oxalis sp, Chlorophytum crassinerve, Psilocaulon sp.). Most plant pieces were collected in winter and spring which is in the moist season. Plantong was consumed the most in the autumn (end of the dry season). Collected plant pieces were multifunctional. Most, 68%, of plant pieces carried back to the lodge were eaten green before they became plantong,Of the plant pieces that were collected, 12% were later eaten as plantong, 11% was incorporated into the structure of the lodge, while the fate of 9% was unknown. Experiments in captivity showed that bush Karoo rats indeed eat plantong. Plantong was not stored for months (from moist to dry season), but mostly days to weeks. Thus, I regard plantong as a food source derived from leftover food during collection and consumption but not as stored food for later consumption during harsh conditions.
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    Assessment of disposal methods of construction and demolition waste: A case study of south-eastern industrial and residential areas in Johannesburg, South Africa
    (University of the Witwatersrand, Johannesburg, 2024) Jager, Vasti de; Kubanza, Nzalalemba Serge
    In a world where all strive for further development, construction and demolition play alarge role in that process. The waste generated in construction and demolition projects is of great magnitude and needs to be dealt with and disposed of appropriately, however, is this truly the case? Gauteng is a province where landfills are easily accessible and a cheap disposal option. This study set out to assess disposal methods of construction and demolition waste in south-east Johannesburg, South Africa. Landfills and recycling were the prevalent disposal methods, and these were compared to other countries’ disposal methods. Policy and legislation regarding solid waste management were analysed and a gap between written documents and implementation was identified. The question of sustainability also played a role in the synthesis of the study
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    Monitoring and evaluating urban land use land cover change using machine learning classification techniques: a case study of Polokwane municipality
    (University of the Witwatersrand, Johannesburg, 2023) Funani, Tshivhase; Mhangara, Paida
    Remote sensing is one of the tools which is very important to produce Land use and land cover maps through the process of image classification. Image classification requires quality multispectral imagery and secondary data, a precise classification technique, and user experience skill. Remote sensing and GIS were used to identify and map land-use/land-cover in the study region. Big Data issues arise when classifying a huge number of satellite images and features, which is a very intensive process. This study primarily uses GEE to evaluate the two classifiers, Support Vector Machine, and gradient boosting, using multi-temporal Landsat-8 images, and to assess their performance while accounting for the impact of data dimension, sample size, and quality. Land use/Landcover (LULC) classification, accuracy assessment, and landscape metrics comprise this study. Gradient Tree Boost and SVM algorithms were used in 2008, 2013, 2017, and 2022. Google Earth Engine was used for supervised classification. The results of change detection showed that urbanization has occurred and most of the encroachments were on agricultural land. In this study, XG boost, and support vector machine (SVM)) were used and compared for image classification to oversight spatio-temporal land use changes in Polokwane Municipality. The Google Earth Engine has been utilized to pre-process the Landsat imagery, and then upload it for classification. Each classification method was evaluated using field observations and high-resolution Google Earth imagery. LULC changes were assessed, utilizing Geographic Information System (GIS) techniques, as well as the dynamics of change in LULCC were analysed using landscape matrix analysis over the last 15 years in four different periods: 2008–2013, 2018 and 2022. The results showed that XGBoost performed better than SVM both in overall accuracies and Kappa statistics as well as F-scores and the ratio of Z-score. The overall accuracy of gradient boosting in 2008 was 0.82, while SVM showed results of 0.82 overall accuracy and kappa statistics of 0.69. The average F-score for SVM in 2008 was from 0.58- 1.00, in 2013 an average of 0.86-0.97, and in 2022 it was 0.76. Z values were not statistically significant as all values were below the z score of 1.96. The ratios for the two classifiers were also taken to know which classifier performs the best. The results showed 212:212 which indicates that during 2008 SVM and XG boost performed the same way as they classified the same number of cases. During 2013 the ratio was 345:312 which shows that XGBoost performed better than SVM. The results of 2017 show 374:316 which shows that XGBoost performed better than SVM. Lastly, in 2022 the ratio was 298:277 which shows that XGBoost performed better than SVM. Overall zscores result show that XGBoost performs better than SVM. Overall, this study offers useful insight into LULC changes that might aid shareholders and decision makers in making informed decisions about controlling land use changes and urban growth
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    The effects of concurrent extreme temperature and water deficit on the phytochemical profile and phytopharmacological activities in Portulacaria afra Jacq
    (University of the Witwatersrand, Johannesburg, 2024) Adeleye, Oluwafunbi Christianah; Risenga, Ida
    In nature, all plants are exposed to stress which are mostly biotic and abiotic stress factors. Previous studies have demonstrated the impact of various abiotic stress factors on the production of secondary metabolites in therapeutic plants. Plant responses to stressors brought on by a combination of antagonistic abiotic factors have shown to be phenomenal compared to when plants are exposed to single a factor. According to latest climate change models, it is believed that plants would suffer unique or demanding concurrent abiotic stresses in the years to come. South Africa has been experiencing increasing temperatures over last 40 years and being regarded as a climate change ‘hot spot’ by the Intergovernmental Panel on Climate Change (IPCC). Therefore, it is critical to conduct research on the impact of climate change on the bioactive compounds in therapeutic plants. The overarching aim of this study was to establish and scientifically document, for the first time to our knowledge, the phytochemical profile, medicinal properties and phytopharmacological attributes of Portulacaria afra, a widely renowned medicinal plant used for treating several skin conditions and oral infections which also includes the assessment of the effects of concurrent extreme temperatures and water deficit/drought on species’ biological activities. In this study, South African Portulacaria afra plants were selected from healthy parent plants and propagated from cuttings. Samples were allowed to grow and establish a root system in the greenhouse for up to three months. After three months, 180 potted plant samples were exposed to treatments and not watered for up to 144 hours (6 days), and 45 control samples were placed under 25oC (ambient) and watered every second day with 500ml of water. The control samples were kept at 25°C maximum night-time temperature (7pm to 5am) and 27°C maximum day time temperature. Plants undergoing treatment (coded as treatment A, B, C, D) were treated as follows: A and B were exposed to 0/10ºC (night/day) and 5/15ºC (night/day) respectively, while C and D were exposed 20/40°C (night/day) and 35/45°C (night/day), respectively. Five potted plants were harvested in each treatment three times for up to 6 days (144hrs). Sample harvesting was scheduled episodically every 48hrs (48, 96, 144) and were then airdried under 40°C for 2 to 3 days. The aqueous (water) extracts at a temperature of 60°C, alongside methanol, n-hexane, and ethyl acetate extracts were derived from the leaves, stems, and roots. The extracts were then used to investigate the phytochemical composition, antibacterial efficacy, antioxidant capacity and antidiabetic potential. The qualitative phytochemical screening encompassed the preliminary assessment of saponins, flavonoids, glycosides, quinones, phenols, terpenoids, steroids, phytosteroids, volatile oil, carbohydrates, amino acids, and coumarins. The quantitative analyses were performed to determine the total phenolic content (TPC), total flavonoid content (TFC), while the antioxidant assays were performed to determine the reducing, scavenging and chelating abilities against DPPH, H2O2 and metal (Iron) chelating. The antibacterial activities against gram-negative Escherichia coli and gram-positive Staphylococcus aureus, Streptomyces griseus were assessed through agar well diffusion assay. The antidiabetic potential was evaluated using In vitro inhibitory α-amylase assay. Chemical profiling of various extracts from the leaves, stems, and roots of P. afra was conducted to identify and quantify some secondary metabolites. The methanolic leaf extracts exhibited a notable presence of quinones, phenols, steroids, and coumarins, whereas the aqueous leaf extracts contained moderate presence of saponins, terpenoids, quinones, and coumarins. Ethyl acetate leaf extracts were characterized by a strong presence of tannins and a moderate presence of phytosteroids. Conversely, n-hexane leaf extracts showed considerable saponin levels, moderate tannins, and terpenoids. Significantly strong presence of secondary metabolites was observed in methanolic stem extracts, particularly terpenoids, steroids, phenols, and coumarins. Notably, coumarins, known for their anticancer properties, were prominently present in methanolic leaf and stem extracts, with a moderate presence in root extracts, hinting at potential pharmaceutical applications and future roles in public health. Aqueous stem extracts exhibited strong glycoside presence, while ethyl acetate and n-hexane stem extracts exhibited few fewer secondary metabolite groups, ranging from moderate to weak presence. Distinctive chemical profiles were observed in root extracts, with ethyl acetate extracts showing significantly stronger quinone presence. Methanolic root extracts displayed moderate presence of coumarins and glycosides, whereas aqueous root extracts showed a low glycoside presence. The highest total phenolic contents (TPCs) and total flavonoid contents (TFCs) were found in methanol stem extracts and aqueous root extracts, respectively. Notably, aqueous root extracts exhibited the highest TPC and TFC among all root extracts. Antibacterial activity assays showed a wide range on inhibitory effects of n-hexane extracts from leaf, stem, and root against test microorganisms. Ethyl acetate leaf extracts demonstrated considerable inhibitory efficacy against Staphylococcus aureus, while methanolic extracts showed zero zone of inhibition. Aqueous root extracts showed strong antibacterial activity against Staphylococcus aureus, whereas other extracts showed no significant activity. Inhibition zones ranged from 13 to 24 mm for the plant extracts. The assessment of antioxidant potential through DPPH, H₂O₂ scavenging, and metal chelating assays showed varied activity among extracts. Ethyl acetate root extracts showed the strongest H₂O₂ scavenging activity, while aqueous stem extracts showed the strongest antioxidant activity against DPPH radicals. Aqueous and n-hexane root extracts showed the strongest metal chelating ability. The in vitro antidiabetic activity showed that all plant parts were active against α-amylase, with the highest inhibitory action recorded from the methanolic leaf extracts, followed by the methanolic root extracts
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    Factors limiting karoo shrub populations in the Nama-Karoo
    (University of the Witwatersrand, Johannesburg, 2023-09) Hebbelmann, Lisa; O’Connor, Tim; Witkowski, Ed
    The Nama-Karoo Biome occupies the central and western region of South Africa. Nama-Karoo shrublands are dominated by karoo dwarf shrubs and in the wetter eastern regions annual and perennial grasses are interspersed among shrubs. Rainfall and grazing are the main drivers of vegetation change in the Nama-Karoo. Stocking rate, season of grazing and animal type are important influences on vegetation community composition, and rainfall determines shrub growth and the abundance of grass. More recently, fire has been identified as a driver of vegetation change. Understanding vegetation change is important for land managers in the Nama-Karoo as changes in karoo shrub communities have implications for forage availability. This thesis is focused on understanding how grazing, fire, grass-shrub competition, and soil moisture and temperature affect the composition, structure, growth and phenology of karoo shrubs in the Eastern Upper Karoo. Livestock type and stocking rate vary greatly over the greater Nama-Karoo area. Plant composition data from the Afrikaner-Hereford grazing trial at Grootfontein show that grazing has a strong legacy effect on Karoo rangelands and grazing-induced compositional and structural changes are clear many years after grazing has ceased. Different sheep breeds affect karoo shrub communities in different ways. There was a severe negative effect of heavy stocking with dorper sheep on karoo shrub communities and their structure; the impacts of merino sheep were significant but less severe. Effects of animal type and stocking rate were not consistent across the sites but appear to be dependent on edaphic factors. Soil physical properties appear to play an important part in determining the long term effects of grazing on karoo shrub communities. Fire is a novel disturbance in the Karoo. Nine sites which had experienced fire for the first time in history were studied for the effect of fire on the composition and structure of their shrub communities. The immediate effect of fire on karoo shrub communities was severe and the legacy effect of fire was significant. The nature of recovery after a fire depended on the abundance of fire-intolerant obligate reseeder shrubs that are killed by fire compared to the abundance of fire-tolerant resprouting shrubs. Rainfall and grazing management were important determinants of shrub recovery after a fire. As rainfall increases, grass biomass is predicted to increase, and this may result in fire becoming a more frequent occurrence in the Nama-Karoo. During periods of high rainfall, the biomass of grass in the Nama-Karoo increased at the expense of shrub cover. The effect of grass-shrub competition on shrub growth and phenology is not adequately explained by existing hypotheses of niche separation and succession. A field experiment which examined the growth and phenology of karoo shrubs in the presence of grass (unclipped control) and the absence of grass (grass removed by clipping) did not provide evidence of a competitive effect from grass on shrub growth or phenology. It did, however, provide valuable insight into how shrub growth and phenology are influenced by soil moisture and temperature. Rainfall in the Nama-Karoo is sporadic and a karoo shrub’s ability to utilise soil moisture will determine its ability to grow and successfully recruit. Shrub growth, with or without grasses in the vicinity, was strongly correlated with rainfall, but this was modified by temperature. Similarly, soil moisture and temperature were important for stem growth and phenology. Flowering occurred in response to soil moisture all year round but seeding occurred mainly during summer. Patterns of growth and phenology can be used by land managers to optimise shrub production while allowing phenological processes to take place. The findings of this study provide further insight into how grazing, fire, competition, soil moisture and temperature influence shrub populations and their structure, and these are discussed in the context of an existing state-and-transition model for the Eastern Upper Karoo. The importance of animal type and stocking rate, and how they vary across Nama-Karoo landscapes, is highlighted. The legacy of both grazing and fire reveal that the transitions between the alternate states of Nama-Karoo shrublands are not easily reversed in a management timeframe. It may take decades for a community changed by grazing or fire, acting independently or in conjunction, to return to their former state even under benign management. These findings along with a description of how soil moisture and temperature influence shrub growth and phenology have been used to make ecologically sound recommendations for the management of Nama-Karoo rangelands. Future research is needed to improve our understanding of how edaphic factors limit and influence karoo shrub populations across a variety of karoo landscapes, how repeat fires might change karoo vegetation, and whether temperature and the presence of pollinators influences shrub seed set. An assessment of the economic implications of vegetation change in the Nama-karoo is warranted so that management recommendations can be made with both ecology and economy in mind.
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    Isolation and characterisation of entomopathogenic fungi
    (University of the Witwatersrand, Johannesburg, 2024) Kwinda, Fhatani; Lephoto, Tiisetso E.
    The purpose of the study was to isolate and identify fungal isolates in soil samples, followed by virulence characterisation to study their effectiveness in controlling insect pests using Tenebrio molitor as our model organism. Lastly, a combination study was conducted to evaluate the effectiveness of the joint use of two entomopathogenic microorganisms. For isolation, T. molitor was used as bait then the isolated fungal isolates were identified using molecular and morphological characterisation. Morphological characterisation included macroscopic (fungal cultures) and microscopic (conidia shape and size) analysis while molecular characterisations included extraction of DNA, amplification of the internal transcribed spacer region and sequence alignment. Once identification was done, virulence was assessed through in-vitro virulence parameter like vegetative growth and in-vivo assessment where bioassays were done against T. molitor. Lastly, entomopathogenic fungi were combined with Cruznema sp. NTM-2021 in a soil assay. From the study, two of the five isolates were identified as entomopathogenic fungi, Metarhizium anisopliae ARSEF 7487. M. anisopliae had the slowest vegetative growth but was the highest in virulence. When used for a single application in a soil environment it reaches 97.8% mortality and its combination with Cruznema sp. NTM-2021 resulted in a 57.8% mortality and an additive interaction. In conclusion, M. anisopliae used alone was effective in its control of T. molitor