The use of the cochineal insect, Dactylopius tomentosus Lamarck, as a biological control agent for the invasive alien thistle cholla, Cylindropuntia pallida (Rose) F.M. Knuth in South Africa
- Authors: Zozo, Ekhona
- Date: 2022-04-06
- Subjects: Cochineal insect , Thistles Biological control South Africa , Alien plants South Africa , Invasive plants South Africa , Cactus South Africa , Biological assay
- Language: English
- Type: Master's thesis , text
- Identifier: http://hdl.handle.net/10962/233823 , vital:50131
- Description: Cylindropuntia pallida (Rose) F.M. Knuth (Cactaceae) is an invasive alien plant in South Africa indigenous in the southern U.S.A. and Mexico. Large infestations of this species can be found in the Kalahari and arid Karoo regions of South Africa, which is also present in Namibia. Because it is a very spiny cactus, dense infestations have a negative impact on agriculture and natural ecosystems. This cactus has become naturalised to the extent that eradication is impossible and the negative impacts are steadily increasing due to its increasing distribution and density. The cochineal insect, Dactylopius tomentosus Lamarck (Dactylopiidae), is native in Mexico and parts of North America, such as Texas, Arizona, and New Mexico in the U.S.A. This cochineal species is highly specialized and associated only with Cylindropuntia species, a group of cacti that are primarily restricted to the same areas. It has been introduced into Australia and South Africa as a biological control agent to control various invasive alien Cylindropuntia species. This cochineal species has several biotypes specific to certain Cylindropuntia species hosts. A biotype is a clade that cannot be differentiated morphologically from others but has different host ranges and impacts depending on the host plant species. Included amongst biotypes that have been released in South Africa are D. tomentosus ‘imbricata’ and D. tomentosus ‘cholla’ for the biological control of Cylindropuntia imbricata (Haw.) F.M. Knuth (Cactaceae) and Cylindropuntia fulgida (Engelmann) F.M. Knuth var. mamillata (Schott ex Engelmann) Backeb. (Cactaceae), respectively. These biotypes have resulted in both host plants being under substantial control in South Africa. The first part of this thesis evaluated which of the two D. tomentosus biotypes already in use in South Africa could be an effective biological control agent for C. pallida. This was done by assessing the fitness of the cochineals on the three cactus species and assessing the impact that each of the cochineals has on each of the target weed species. Should these biotypes prove ineffective, there is a third biotype, namely D. tomentosus ‘californica var. parkerii’, which researchers in Australia have worked on and have found to be suitably host-specific for release in Australia and suitably damaging to C. pallida in that country. Therefore, this new biotype could be released in South Africa if it is required. Sexually compatible biological control agents, especially those closely related and occurring in close spatial proximity to one another, may interbreed and the impacts of this hybridisation are difficult to predict. It is important to understand the outcomes of the hybridisation of cochineal because it can affect the impact of the biological control agents and thus the control of the target weed. The second part of this thesis investigated the impacts of the hybridisation of the two cochineal biotypes by assessing the damage the agents would have on the target weed, and on C. imbricata and C. fulgida var. mamillata, in the presence of one or both cochineal biotypes. The ‘cholla’ biotype performed better on C. pallida than the ‘imbricata’ biotype, but neither biotype could control C. pallida to an extent similar to the control they provide for their respective target weeds, C. imbricata and C. fulgida var. mamillata. Both the ‘cholla’ biotype and hybrids of the two biotypes of cochineal were effective at killing C. pallida when both C. imbricata and C. fulgida var. mamillata were also present. This suggests that the ‘cholla’ or hybrids may be effective at controlling C. pallida when either C. imbricata or C. fulgida var. mamillata are also present in the field due to the high population density of cochineal that results under these circumstances. There are, however, many C. pallida infestations in South Africa where the plant is problematic and is isolated from other Cylindropuntia species, and these populations are unlikely to be controlled by the ‘cholla’ biotype or the hybrids. Neither of the cochineal biotypes that are used for biological control in South Africa are suitably damaging to C. pallida to warrant their use as biological control agents for this species. Dactylopius tomentosus ‘californica var. parkerii’ is therefore recommended for release based on its host-specificity and impact to C. pallida in Australia. , Thesis (PhD) -- Faculty of Science, Zoology and Entomology, 2022
- Full Text:
- Date Issued: 2022-04-06
- Authors: Zozo, Ekhona
- Date: 2022-04-06
- Subjects: Cochineal insect , Thistles Biological control South Africa , Alien plants South Africa , Invasive plants South Africa , Cactus South Africa , Biological assay
- Language: English
- Type: Master's thesis , text
- Identifier: http://hdl.handle.net/10962/233823 , vital:50131
- Description: Cylindropuntia pallida (Rose) F.M. Knuth (Cactaceae) is an invasive alien plant in South Africa indigenous in the southern U.S.A. and Mexico. Large infestations of this species can be found in the Kalahari and arid Karoo regions of South Africa, which is also present in Namibia. Because it is a very spiny cactus, dense infestations have a negative impact on agriculture and natural ecosystems. This cactus has become naturalised to the extent that eradication is impossible and the negative impacts are steadily increasing due to its increasing distribution and density. The cochineal insect, Dactylopius tomentosus Lamarck (Dactylopiidae), is native in Mexico and parts of North America, such as Texas, Arizona, and New Mexico in the U.S.A. This cochineal species is highly specialized and associated only with Cylindropuntia species, a group of cacti that are primarily restricted to the same areas. It has been introduced into Australia and South Africa as a biological control agent to control various invasive alien Cylindropuntia species. This cochineal species has several biotypes specific to certain Cylindropuntia species hosts. A biotype is a clade that cannot be differentiated morphologically from others but has different host ranges and impacts depending on the host plant species. Included amongst biotypes that have been released in South Africa are D. tomentosus ‘imbricata’ and D. tomentosus ‘cholla’ for the biological control of Cylindropuntia imbricata (Haw.) F.M. Knuth (Cactaceae) and Cylindropuntia fulgida (Engelmann) F.M. Knuth var. mamillata (Schott ex Engelmann) Backeb. (Cactaceae), respectively. These biotypes have resulted in both host plants being under substantial control in South Africa. The first part of this thesis evaluated which of the two D. tomentosus biotypes already in use in South Africa could be an effective biological control agent for C. pallida. This was done by assessing the fitness of the cochineals on the three cactus species and assessing the impact that each of the cochineals has on each of the target weed species. Should these biotypes prove ineffective, there is a third biotype, namely D. tomentosus ‘californica var. parkerii’, which researchers in Australia have worked on and have found to be suitably host-specific for release in Australia and suitably damaging to C. pallida in that country. Therefore, this new biotype could be released in South Africa if it is required. Sexually compatible biological control agents, especially those closely related and occurring in close spatial proximity to one another, may interbreed and the impacts of this hybridisation are difficult to predict. It is important to understand the outcomes of the hybridisation of cochineal because it can affect the impact of the biological control agents and thus the control of the target weed. The second part of this thesis investigated the impacts of the hybridisation of the two cochineal biotypes by assessing the damage the agents would have on the target weed, and on C. imbricata and C. fulgida var. mamillata, in the presence of one or both cochineal biotypes. The ‘cholla’ biotype performed better on C. pallida than the ‘imbricata’ biotype, but neither biotype could control C. pallida to an extent similar to the control they provide for their respective target weeds, C. imbricata and C. fulgida var. mamillata. Both the ‘cholla’ biotype and hybrids of the two biotypes of cochineal were effective at killing C. pallida when both C. imbricata and C. fulgida var. mamillata were also present. This suggests that the ‘cholla’ or hybrids may be effective at controlling C. pallida when either C. imbricata or C. fulgida var. mamillata are also present in the field due to the high population density of cochineal that results under these circumstances. There are, however, many C. pallida infestations in South Africa where the plant is problematic and is isolated from other Cylindropuntia species, and these populations are unlikely to be controlled by the ‘cholla’ biotype or the hybrids. Neither of the cochineal biotypes that are used for biological control in South Africa are suitably damaging to C. pallida to warrant their use as biological control agents for this species. Dactylopius tomentosus ‘californica var. parkerii’ is therefore recommended for release based on its host-specificity and impact to C. pallida in Australia. , Thesis (PhD) -- Faculty of Science, Zoology and Entomology, 2022
- Full Text:
- Date Issued: 2022-04-06
Synthetic analogues of marine bisindole alkaloids as potent selective inhibitors of MRSA pyruvate kinase
- Veale, Clinton Gareth Lancaster
- Authors: Veale, Clinton Gareth Lancaster
- Date: 2014 , 2014-04-02
- Subjects: Alkaloids , Pyruvate kinase , Staphylococcus aureus , Antibiotics , Sponges -- South Africa , Imidazoles , Biological assay , Antibacterial agents
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4563 , http://hdl.handle.net/10962/d1020893
- Description: Globally, methicillin resistant Staphylococcus aureus (MRSA) has become increasingly difficult to manage in the clinic and new antibiotics are required. The structure activity relationship (SAR) study presented in this thesis forms part of an international collaborative effort to identify potent and selective inhibitors of an MRSA pyruvate kinase (PK) enzyme target. In earlier work the known marine natural product bromodeoxytopsentin (1.6), isolated from a South African marine sponge Topsentia pachastrelloides, exhibited selective and significant inhibition of MRSA PK (IC₅₀ 60 nM). Accordingly bromodeoxytopsentin provided the initial chemical scaffold around which our SAR study was developed. Following a comprehensive introduction, providing the necessary background to the research described in subsequent Chapters, this thesis has been divided into three major parts. Part one (Chapter 2) documents the synthesis of two natural imidazole containing topsentin analogues 1.40, 1.46, five new synthetic analogues 1.58—1.61, 2.104. In the process we developed a new method for the synthesis of topsentin derivatives via selenium dioxide mediated oxidation of N-Boc protected 3-acetylindoles to yield glyoxal intermediates which were subsequently cyclized and deprotected to yield the desired products. Interestingly we were able to demonstrate a delicate relationship between the relative equivalents of selenium dioxide and water used during the oxidation step, careful manipulation of which was required to prevent the uncontrolled formation of side products. Synthetic compounds 1.40, 1.46, 1.58—1.61 were found to be potent inhibitors of MRSA PK (IC₅₀ 238, 2.1, 23, 1.4, 6.3 and 3.2 nM respectively) with 1000-10000 fold selectivity for MRSA PK over four human orthologs. In the second part of this thesis (Chapter 3) we report the successful synthesis of a cohort of previously unknown thiazole containing bisindole topsentin analogues 1.62—1.68 via a Hantzsch thiazole synthesis. Bioassay results revealed that these compounds were only moderate inhibitors of MRSA PK (IC₅₀ 5.1—20 μM) which suggested that inhibitory activity was significantly reduced upon substitution of the central imidazole ring of topsentin type analogues with a thiazole type ring. In addition in Chapter 3 we describe unsuccessful attempts to regiospecifically synthesize oxazole and imidazole topsentin analogues through a similar Hantzsch method. As a consequence of our efforts in this regard we investigated three key reactions in depth, namely the synthesis of 2.2, 3.38, 3.40, 3.41 via α-bromination of 3-acetylindole and the synthesis of indolyl-3-carbonylnitriles 2.13, 3.45—3.47 and α-oxo-1H-indole-3-thioacetamides 3.48—3.51. The investigation of the latter led to the isolation and elucidation of two anomalous N,N-dimethyl-1H-indole-3-carboxamides 3.52 and 3.53. Finally the third part of this thesis (Chapter 4) deals with in silico assessment of the binding of both the imidazole and thiazole containing bisindole alkaloids to the MRSA PK protein which initially guided our SAR studies. In this chapter we reveal that there appears to be no correlation between in silico binding predictions and in vitro MRSA PK inhibitory bioassay data. Superficially it seems that binding energy as determined by the docking program used for these studies correlated with the size of the indole substituents and did not reflect IC₅₀ MRSA PK inhibitory data. Although this led us to computationally explore possible alternative binding sites no clear alternative has been identified.
- Full Text:
- Date Issued: 2014
- Authors: Veale, Clinton Gareth Lancaster
- Date: 2014 , 2014-04-02
- Subjects: Alkaloids , Pyruvate kinase , Staphylococcus aureus , Antibiotics , Sponges -- South Africa , Imidazoles , Biological assay , Antibacterial agents
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4563 , http://hdl.handle.net/10962/d1020893
- Description: Globally, methicillin resistant Staphylococcus aureus (MRSA) has become increasingly difficult to manage in the clinic and new antibiotics are required. The structure activity relationship (SAR) study presented in this thesis forms part of an international collaborative effort to identify potent and selective inhibitors of an MRSA pyruvate kinase (PK) enzyme target. In earlier work the known marine natural product bromodeoxytopsentin (1.6), isolated from a South African marine sponge Topsentia pachastrelloides, exhibited selective and significant inhibition of MRSA PK (IC₅₀ 60 nM). Accordingly bromodeoxytopsentin provided the initial chemical scaffold around which our SAR study was developed. Following a comprehensive introduction, providing the necessary background to the research described in subsequent Chapters, this thesis has been divided into three major parts. Part one (Chapter 2) documents the synthesis of two natural imidazole containing topsentin analogues 1.40, 1.46, five new synthetic analogues 1.58—1.61, 2.104. In the process we developed a new method for the synthesis of topsentin derivatives via selenium dioxide mediated oxidation of N-Boc protected 3-acetylindoles to yield glyoxal intermediates which were subsequently cyclized and deprotected to yield the desired products. Interestingly we were able to demonstrate a delicate relationship between the relative equivalents of selenium dioxide and water used during the oxidation step, careful manipulation of which was required to prevent the uncontrolled formation of side products. Synthetic compounds 1.40, 1.46, 1.58—1.61 were found to be potent inhibitors of MRSA PK (IC₅₀ 238, 2.1, 23, 1.4, 6.3 and 3.2 nM respectively) with 1000-10000 fold selectivity for MRSA PK over four human orthologs. In the second part of this thesis (Chapter 3) we report the successful synthesis of a cohort of previously unknown thiazole containing bisindole topsentin analogues 1.62—1.68 via a Hantzsch thiazole synthesis. Bioassay results revealed that these compounds were only moderate inhibitors of MRSA PK (IC₅₀ 5.1—20 μM) which suggested that inhibitory activity was significantly reduced upon substitution of the central imidazole ring of topsentin type analogues with a thiazole type ring. In addition in Chapter 3 we describe unsuccessful attempts to regiospecifically synthesize oxazole and imidazole topsentin analogues through a similar Hantzsch method. As a consequence of our efforts in this regard we investigated three key reactions in depth, namely the synthesis of 2.2, 3.38, 3.40, 3.41 via α-bromination of 3-acetylindole and the synthesis of indolyl-3-carbonylnitriles 2.13, 3.45—3.47 and α-oxo-1H-indole-3-thioacetamides 3.48—3.51. The investigation of the latter led to the isolation and elucidation of two anomalous N,N-dimethyl-1H-indole-3-carboxamides 3.52 and 3.53. Finally the third part of this thesis (Chapter 4) deals with in silico assessment of the binding of both the imidazole and thiazole containing bisindole alkaloids to the MRSA PK protein which initially guided our SAR studies. In this chapter we reveal that there appears to be no correlation between in silico binding predictions and in vitro MRSA PK inhibitory bioassay data. Superficially it seems that binding energy as determined by the docking program used for these studies correlated with the size of the indole substituents and did not reflect IC₅₀ MRSA PK inhibitory data. Although this led us to computationally explore possible alternative binding sites no clear alternative has been identified.
- Full Text:
- Date Issued: 2014
Rapid enzymatic detection of organophosphorous and carbamate pesticides in water
- Authors: Mwila, Katayi
- Date: 2012
- Subjects: Organophosphorus compounds , Carbamates , Water -- Pesticide content -- South Africa -- Eastern Cape , Water quality biological assessment -- South Africa -- Eastern Cape , Water quality management -- South Africa -- Eastern Cape , Pesticides -- Toxicology -- South Africa -- Eastern Cape , Biological assay , Acetylcholinesterase , Parathion , Aldicarb , Carbaryl , Carbofuran , Nitrophenols
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4024 , http://hdl.handle.net/10962/d1004084 , Organophosphorus compounds , Carbamates , Water -- Pesticide content -- South Africa -- Eastern Cape , Water quality biological assessment -- South Africa -- Eastern Cape , Water quality management -- South Africa -- Eastern Cape , Pesticides -- Toxicology -- South Africa -- Eastern Cape , Biological assay , Acetylcholinesterase , Parathion , Aldicarb , Carbaryl , Carbofuran , Nitrophenols
- Description: The increased use of pesticides has resulted in a corresponding increase in concern for the effect they may have on the health of humans and other non-target organisms. The two main areas of concern are the toxicological effects that mixtures of pesticides may have as well as the endocrine disrupting effects. Although the individual pesticides may be present at concentrations below the levels deemed to be detrimental to health, it has been argued that their combined effect may still result in elevated health risks. Another important aspect of pesticide risk assessment requires a consideration of the breakdown products of pesticides and their effect on human health. There has been very little research into the effects of degradation products and this issue should be addressed as these could potentially pose a higher risk than their parent compounds. One of the most important bio-markers available for use is the ubiquitous enzyme acetylcholinesterase (AChE). This enzyme is responsible for one of the most important functions in the body; namely nerve impulse transmission, upon which all life depends. The inhibition of this enzyme indicates toxicity and as a subsequence, a threat to the organism’s well-being. Bioassays have also recently been developed to test chemicals for endocrine disrupting effects. These tests rely on a dose response equivalent to that of the most potent well known estrogen 17-β estradiol. Any chemical that has a measurable response is deemed to display endocrine disrupting effects. This first aim of this study was to investigate the toxicological and endocrine disrupting effects of three organophosphorus pesticides; aldicarb, parathion and demeton-S-methyl, in addition to two breakdown products; aminophenol and p-nitrophenol. Two carbamate pesticides; carbaryl and carbofuran were also analysed. The toxicological effects of mixtures of the parent pesticide compounds were tested to assess if any antagonistic, additive or synergistic effects were observed. This data was then used in conjunction with an artificial neural network to assess if individual pesticides could be distinguished from mixtures of pesticides. A final objective was to sample various Eastern Cape water sources, utilising the enzymatic assay to determine the presence of any of these pesticides in these samples. There were several conclusions drawn from this study. AChE was successfully used as an assay to test the toxicity of the pesticides under investigation, based on their inhibition of this enzyme. An important factor for consideration throughout the study was the need to establish basal and monitor AChE activity (i.e. the need to monitor AChE activity in the absence of any pesticide). This ensured accurate comparison of the results obtained. It was found that demeton-S-methyl was the most potent of these pesticides followed by carbaryl, parathion, aldicarb and finally carbofuran, and that carbofuran could potentiate AChE. The results indicated that pesticide mixtures generally exhibited an additive inhibitory effect on AChE, although at some concentrations of pesticides, synergistic and antagonistic effects were noted. From the data using mixtures of pesticides, a feed forward neural network was created that was successfully able to distinguish individual pesticides from mixtures within its training parameters. None of the pesticides tested displayed endocrine disrupting properties in the Yeast Estrogen Screen (YES), T47D-KBluc and MDA-kb2 bio-assays. Other studies reported mixed results in this regard and thus no final conclusions could be drawn. The Blaauwkrantz River, Kariega River, Sundays River, Swartkops River and Kowie River were all tested for pesticides and although positive results were recorded, conventional methods indicated that there were no pesticides in the rivers. There were, however, trace metals present which are known to inhibit AChE, thus causing a false positive result. These results indicated that AChE can be used as a high throughput initial pre-screening tool, but that it cannot serve as a substitute for more accurate conventional testing methods.
- Full Text:
- Date Issued: 2012
- Authors: Mwila, Katayi
- Date: 2012
- Subjects: Organophosphorus compounds , Carbamates , Water -- Pesticide content -- South Africa -- Eastern Cape , Water quality biological assessment -- South Africa -- Eastern Cape , Water quality management -- South Africa -- Eastern Cape , Pesticides -- Toxicology -- South Africa -- Eastern Cape , Biological assay , Acetylcholinesterase , Parathion , Aldicarb , Carbaryl , Carbofuran , Nitrophenols
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4024 , http://hdl.handle.net/10962/d1004084 , Organophosphorus compounds , Carbamates , Water -- Pesticide content -- South Africa -- Eastern Cape , Water quality biological assessment -- South Africa -- Eastern Cape , Water quality management -- South Africa -- Eastern Cape , Pesticides -- Toxicology -- South Africa -- Eastern Cape , Biological assay , Acetylcholinesterase , Parathion , Aldicarb , Carbaryl , Carbofuran , Nitrophenols
- Description: The increased use of pesticides has resulted in a corresponding increase in concern for the effect they may have on the health of humans and other non-target organisms. The two main areas of concern are the toxicological effects that mixtures of pesticides may have as well as the endocrine disrupting effects. Although the individual pesticides may be present at concentrations below the levels deemed to be detrimental to health, it has been argued that their combined effect may still result in elevated health risks. Another important aspect of pesticide risk assessment requires a consideration of the breakdown products of pesticides and their effect on human health. There has been very little research into the effects of degradation products and this issue should be addressed as these could potentially pose a higher risk than their parent compounds. One of the most important bio-markers available for use is the ubiquitous enzyme acetylcholinesterase (AChE). This enzyme is responsible for one of the most important functions in the body; namely nerve impulse transmission, upon which all life depends. The inhibition of this enzyme indicates toxicity and as a subsequence, a threat to the organism’s well-being. Bioassays have also recently been developed to test chemicals for endocrine disrupting effects. These tests rely on a dose response equivalent to that of the most potent well known estrogen 17-β estradiol. Any chemical that has a measurable response is deemed to display endocrine disrupting effects. This first aim of this study was to investigate the toxicological and endocrine disrupting effects of three organophosphorus pesticides; aldicarb, parathion and demeton-S-methyl, in addition to two breakdown products; aminophenol and p-nitrophenol. Two carbamate pesticides; carbaryl and carbofuran were also analysed. The toxicological effects of mixtures of the parent pesticide compounds were tested to assess if any antagonistic, additive or synergistic effects were observed. This data was then used in conjunction with an artificial neural network to assess if individual pesticides could be distinguished from mixtures of pesticides. A final objective was to sample various Eastern Cape water sources, utilising the enzymatic assay to determine the presence of any of these pesticides in these samples. There were several conclusions drawn from this study. AChE was successfully used as an assay to test the toxicity of the pesticides under investigation, based on their inhibition of this enzyme. An important factor for consideration throughout the study was the need to establish basal and monitor AChE activity (i.e. the need to monitor AChE activity in the absence of any pesticide). This ensured accurate comparison of the results obtained. It was found that demeton-S-methyl was the most potent of these pesticides followed by carbaryl, parathion, aldicarb and finally carbofuran, and that carbofuran could potentiate AChE. The results indicated that pesticide mixtures generally exhibited an additive inhibitory effect on AChE, although at some concentrations of pesticides, synergistic and antagonistic effects were noted. From the data using mixtures of pesticides, a feed forward neural network was created that was successfully able to distinguish individual pesticides from mixtures within its training parameters. None of the pesticides tested displayed endocrine disrupting properties in the Yeast Estrogen Screen (YES), T47D-KBluc and MDA-kb2 bio-assays. Other studies reported mixed results in this regard and thus no final conclusions could be drawn. The Blaauwkrantz River, Kariega River, Sundays River, Swartkops River and Kowie River were all tested for pesticides and although positive results were recorded, conventional methods indicated that there were no pesticides in the rivers. There were, however, trace metals present which are known to inhibit AChE, thus causing a false positive result. These results indicated that AChE can be used as a high throughput initial pre-screening tool, but that it cannot serve as a substitute for more accurate conventional testing methods.
- Full Text:
- Date Issued: 2012
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