Molecular systematics and biology of two closely related blowflies : Lucilia sericata and Lucilia cuprina
- Authors: Williams, Kirstin Alexa
- Date: 2015
- Subjects: Lucilia sericata , Lucilia cuprina
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5934 , http://hdl.handle.net/10962/d1017812
- Description: The greenbottle blowflies, Lucilia sericata (Meigen, 1826) and Lucilia cuprina (Wiedemann, 1830) (Diptera: Calliphoridae) are very difficult to distinguish on the basis of their external morphology. The literature suggests that these two species may be interbreeding. Sequencing two nuclear (28S rRNA and Period) and one mitochondrial (COI) gene indicated that there has been an ancient hybridization event and that mtDNA of L. sericata has become fixed in a lineage of L. cuprina through mtDNA introgression, possibly involving Wolbachia infection. This has implications for identifications of these species based on mtDNA alone. No study has shown explicitly that hybrids of L. sericata and L. cuprina can be identified morphologically. Morphological characters used to identify L. sericata and L. cuprina were scored and tested using specimens of both species and known hybrids. Discriminant function analysis of the characters successfully separated the specimens into three unambiguous groups – L. sericata, L. cuprina and hybrids. This is the first evidence that hybrids of these two species can be identified from physical characteristics.Lucilia sericata and L. cuprina have medical, veterinary and forensic importance. Knowing their distribution in South Africa would allow more effective management and utilisation of these flies. Their predicted geographic distributions in South Africa were modelled using maximum entropy analysis of selected climatic variables. The most important environmental variables in modelling their distributions were magnitude of monthly rainfall and the magnitude of the monthly maximum temperature for L. sericata, and the seasonal variation in monthly mean humidity and magnitude of monthly rainfall for L. cuprina. Both species have a widespread distribution in South Africa and one therefore cannot identify specimens of these flies by locality of capture alone.Luciliinae is a diverse and geographically widespread subfamily containing four genera - Hemipyrellia, Lucilia, Dyscritomyia and Hypopygiopsis – that all contain parasitic species ranging from saprophages to obligate parasites. The phylogenetic relationships between these genera are unclear. The 28S rRNA, COI and Period genes of 14 species of Lucilia and Hemipyrellia were partially sequenced and analysed together with 11sequences from GenBank and the Barcode of Life Database (BOLD). Lucilia sericata and L. cuprina were shown to be sister-species. Three cases of paraphylly were identified within Lucilia that affects identification of these species using mtDNA alone. Hemipyrellia consistently caused Lucilia to be paraphyletic when it was included in analyses, so Hemipyrellia should be synonymized with Lucilia. The relationships of Dyscritomyia and Hypopygiopsis to Lucilia are unclear and further studies are required. No geographic pattern was found within the different forms of parasitism within this group, but the different degrees of parasitism were phylogenetically clustered.
- Full Text:
- Authors: Williams, Kirstin Alexa
- Date: 2015
- Subjects: Lucilia sericata , Lucilia cuprina
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:5934 , http://hdl.handle.net/10962/d1017812
- Description: The greenbottle blowflies, Lucilia sericata (Meigen, 1826) and Lucilia cuprina (Wiedemann, 1830) (Diptera: Calliphoridae) are very difficult to distinguish on the basis of their external morphology. The literature suggests that these two species may be interbreeding. Sequencing two nuclear (28S rRNA and Period) and one mitochondrial (COI) gene indicated that there has been an ancient hybridization event and that mtDNA of L. sericata has become fixed in a lineage of L. cuprina through mtDNA introgression, possibly involving Wolbachia infection. This has implications for identifications of these species based on mtDNA alone. No study has shown explicitly that hybrids of L. sericata and L. cuprina can be identified morphologically. Morphological characters used to identify L. sericata and L. cuprina were scored and tested using specimens of both species and known hybrids. Discriminant function analysis of the characters successfully separated the specimens into three unambiguous groups – L. sericata, L. cuprina and hybrids. This is the first evidence that hybrids of these two species can be identified from physical characteristics.Lucilia sericata and L. cuprina have medical, veterinary and forensic importance. Knowing their distribution in South Africa would allow more effective management and utilisation of these flies. Their predicted geographic distributions in South Africa were modelled using maximum entropy analysis of selected climatic variables. The most important environmental variables in modelling their distributions were magnitude of monthly rainfall and the magnitude of the monthly maximum temperature for L. sericata, and the seasonal variation in monthly mean humidity and magnitude of monthly rainfall for L. cuprina. Both species have a widespread distribution in South Africa and one therefore cannot identify specimens of these flies by locality of capture alone.Luciliinae is a diverse and geographically widespread subfamily containing four genera - Hemipyrellia, Lucilia, Dyscritomyia and Hypopygiopsis – that all contain parasitic species ranging from saprophages to obligate parasites. The phylogenetic relationships between these genera are unclear. The 28S rRNA, COI and Period genes of 14 species of Lucilia and Hemipyrellia were partially sequenced and analysed together with 11sequences from GenBank and the Barcode of Life Database (BOLD). Lucilia sericata and L. cuprina were shown to be sister-species. Three cases of paraphylly were identified within Lucilia that affects identification of these species using mtDNA alone. Hemipyrellia consistently caused Lucilia to be paraphyletic when it was included in analyses, so Hemipyrellia should be synonymized with Lucilia. The relationships of Dyscritomyia and Hypopygiopsis to Lucilia are unclear and further studies are required. No geographic pattern was found within the different forms of parasitism within this group, but the different degrees of parasitism were phylogenetically clustered.
- Full Text:
Adolescent pregnancy: a community engaged participatory approach to design and implement an educational intervention
- Authors: Siruma, Amanda Tatenda
- Date: 2014
- Language: English
- Type: text , Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/54656 , vital:26597
- Description: Millennium Development Goal (MDG) 5 focuses on improving maternal health, due to global acknowledgment that no woman should have to die as a result of complications during pregnancy and childbirth. Adolescents have an increased risk of maternal death compared with older women. Adolescent pregnancy also poses a threat to the empowerment of young girls by mitigating their physical, educational, social, and economic development. In this context, maternal health promotion strategies which inclusively target adolescents are crucial, not only in improving maternal health outcomes, but also in optimising the overall transition of adolescent girls to adulthood. This study was a first time collaborative partnership of the Faculty of Pharmacy and Community Engagement Office of Rhodes University with the Angus Gillis Foundation (a non-profit community development organisation), and community participants of Glenmore and Ndwayana, two rural communities in the Eastern Cape. The aim of this study was to identify the maternal health issue of most concern to community participants and to design and implement an appropriate educational intervention for a suitable target group. During the baseline phase of this study, ten focus group discussions (FGDs) were conducted with 76 community stakeholders. Semi-structured interviews (SSIs) were conducted with two Sisters-in-Charge from each Primary Health Care (PHC) facility in the study setting. Data on the stock status of World Health Organization (WHO) identified lifesaving priority medicines for women’s health was also collected at both PHCs. Thereafter, pre- and posteducational interventions SSIs with female adolescent participants were conducted. The educational intervention was followed up with the development of a booklet on reproductive health. FGD participants identified adolescent pregnancy as the maternal health issue of most concern. They also highlighted challenges in service delivery of ambulance services for expectant mothers in urgent need of transportation to a referral hospital. A majority of preintervention SSI participants indicated coercion from both younger and older men as a factor influencing early sexual debut amongst adolescent girls in their communities. Despite availability in the PHCs, challenges in accessing contraceptives were highlighted by the participants. Additionally, a number of sexually active adolescent girls defaulted on their next allocated visit to the PHC due to myths regarding use of oral and injectable contraceptives. During the educational intervention sessions, participants recognised knowledge gaps regarding reproductive health issues and the influence of peer pressure as constraining factors in preventing adolescent pregnancy. During the post-intervention phase, participants highlighted that the educational intervention of this study had provided a forum to discuss ways of preventing adolescent pregnancy. The educational booklet developed is intended to serve as a resource tool of the educational programme on prevention of adolescent pregnancy, which is expected to be incorporated into the Angus Gillis Foundation’s existing ‘Positive Health’ Programme. The results of this study show that community-based participatory research facilitated the identification of the maternal health issue of most concern to these communities. Working synergistically with key stakeholders in designing and implementing an educational intervention for preventing adolescent pregnancy provides a good foundation for future up scaling and sustainability of this educational programme.
- Full Text:
- Authors: Siruma, Amanda Tatenda
- Date: 2014
- Language: English
- Type: text , Thesis , Masters , MPharm
- Identifier: http://hdl.handle.net/10962/54656 , vital:26597
- Description: Millennium Development Goal (MDG) 5 focuses on improving maternal health, due to global acknowledgment that no woman should have to die as a result of complications during pregnancy and childbirth. Adolescents have an increased risk of maternal death compared with older women. Adolescent pregnancy also poses a threat to the empowerment of young girls by mitigating their physical, educational, social, and economic development. In this context, maternal health promotion strategies which inclusively target adolescents are crucial, not only in improving maternal health outcomes, but also in optimising the overall transition of adolescent girls to adulthood. This study was a first time collaborative partnership of the Faculty of Pharmacy and Community Engagement Office of Rhodes University with the Angus Gillis Foundation (a non-profit community development organisation), and community participants of Glenmore and Ndwayana, two rural communities in the Eastern Cape. The aim of this study was to identify the maternal health issue of most concern to community participants and to design and implement an appropriate educational intervention for a suitable target group. During the baseline phase of this study, ten focus group discussions (FGDs) were conducted with 76 community stakeholders. Semi-structured interviews (SSIs) were conducted with two Sisters-in-Charge from each Primary Health Care (PHC) facility in the study setting. Data on the stock status of World Health Organization (WHO) identified lifesaving priority medicines for women’s health was also collected at both PHCs. Thereafter, pre- and posteducational interventions SSIs with female adolescent participants were conducted. The educational intervention was followed up with the development of a booklet on reproductive health. FGD participants identified adolescent pregnancy as the maternal health issue of most concern. They also highlighted challenges in service delivery of ambulance services for expectant mothers in urgent need of transportation to a referral hospital. A majority of preintervention SSI participants indicated coercion from both younger and older men as a factor influencing early sexual debut amongst adolescent girls in their communities. Despite availability in the PHCs, challenges in accessing contraceptives were highlighted by the participants. Additionally, a number of sexually active adolescent girls defaulted on their next allocated visit to the PHC due to myths regarding use of oral and injectable contraceptives. During the educational intervention sessions, participants recognised knowledge gaps regarding reproductive health issues and the influence of peer pressure as constraining factors in preventing adolescent pregnancy. During the post-intervention phase, participants highlighted that the educational intervention of this study had provided a forum to discuss ways of preventing adolescent pregnancy. The educational booklet developed is intended to serve as a resource tool of the educational programme on prevention of adolescent pregnancy, which is expected to be incorporated into the Angus Gillis Foundation’s existing ‘Positive Health’ Programme. The results of this study show that community-based participatory research facilitated the identification of the maternal health issue of most concern to these communities. Working synergistically with key stakeholders in designing and implementing an educational intervention for preventing adolescent pregnancy provides a good foundation for future up scaling and sustainability of this educational programme.
- Full Text:
The principle of complementarity : a critical analysis of Article 17 of the Rome Statute from an African perspective
- Authors: Mohami, Thapelo Adelice
- Date: 2014
- Subjects: Complementarity (International law) , Rome Statute of the International Criminal Court (1998 July 17) , International Criminal Court , Criminal jurisdiction -- Africa , International crimes -- Africa , Crimes against humanity -- Africa
- Language: English
- Type: Thesis , Masters , LLM
- Identifier: vital:3720 , http://hdl.handle.net/10962/d1013326
- Description: This thesis attempts to address perennial concerns, mostly raised in some quarters in Africa, pertaining to the development of the complementarity regime established by the Rome Statute of the International Criminal Court. It grapples with a very important question, whether the principle of complementarity, embodied in article 17 of the Rome Statute, was formulated and is being applied by the ICC in a manner that upholds the ideals and theories upon which the regime was founded. The principle of complementarity is designed to mediate the imperatives of State sovereignty and a legitimate international criminal justice system. Essentially, complementarity gives States latitude to try genocide, crimes against humanity, war crimes and aggression nationally, with the ICC only intervening where States are either unable or unwilling to prosecute genuinely. Africa constitutes the biggest regional block of membership to the Rome Statute, however, over the years; support for the ICC on the African continent has waned. It has been argued in some quarters that the ICC is anti-African and that it has interpreted and applied complementarity in a manner that diminishes State sovereignty. The thesis argues that this tension may also be due to textual deficiencies inherent within the Rome Statute, in the provisions that embody this principle. It therefore examines complementarity from a theoretical perspective to provide a comprehensive account of the system contemplated by the drafters of the Rome Statute. In this regard, the thesis argues for expansion of States’ ability at the national level to deal with international crimes without compromising international criminal justice processes or threatening State sovereignty. This is suggested as a way of relieving the tension that has characterised the relationship between African States and the ICC. The thesis further sketches out some of the complexities inherent in the modalities through which the Court may exercise its complementary jurisdiction, particularly within the African continent, given that legal systems in most African countries are particularly weak. It thus dissects the provisions that outline the principle of complementarity in tandem with the Court’s interpretation and application of complementarity in practice. Furthermore, through an exploratory survey of the referral of the Situation in Uganda, and the ICC Prosecutor’s proprio motu investigation of the Situation in Kenya, the thesis illustrates how a positive approach to complementarity can help establish a healthy cooperative synergy between the ICC and States, thereby promoting a functional expeditious criminal justice system. This will go a long way towards assuaging State’s fears that the ICC merely pays lip service to complementarity and arbitrarily supersedes national jurisdiction.
- Full Text:
- Authors: Mohami, Thapelo Adelice
- Date: 2014
- Subjects: Complementarity (International law) , Rome Statute of the International Criminal Court (1998 July 17) , International Criminal Court , Criminal jurisdiction -- Africa , International crimes -- Africa , Crimes against humanity -- Africa
- Language: English
- Type: Thesis , Masters , LLM
- Identifier: vital:3720 , http://hdl.handle.net/10962/d1013326
- Description: This thesis attempts to address perennial concerns, mostly raised in some quarters in Africa, pertaining to the development of the complementarity regime established by the Rome Statute of the International Criminal Court. It grapples with a very important question, whether the principle of complementarity, embodied in article 17 of the Rome Statute, was formulated and is being applied by the ICC in a manner that upholds the ideals and theories upon which the regime was founded. The principle of complementarity is designed to mediate the imperatives of State sovereignty and a legitimate international criminal justice system. Essentially, complementarity gives States latitude to try genocide, crimes against humanity, war crimes and aggression nationally, with the ICC only intervening where States are either unable or unwilling to prosecute genuinely. Africa constitutes the biggest regional block of membership to the Rome Statute, however, over the years; support for the ICC on the African continent has waned. It has been argued in some quarters that the ICC is anti-African and that it has interpreted and applied complementarity in a manner that diminishes State sovereignty. The thesis argues that this tension may also be due to textual deficiencies inherent within the Rome Statute, in the provisions that embody this principle. It therefore examines complementarity from a theoretical perspective to provide a comprehensive account of the system contemplated by the drafters of the Rome Statute. In this regard, the thesis argues for expansion of States’ ability at the national level to deal with international crimes without compromising international criminal justice processes or threatening State sovereignty. This is suggested as a way of relieving the tension that has characterised the relationship between African States and the ICC. The thesis further sketches out some of the complexities inherent in the modalities through which the Court may exercise its complementary jurisdiction, particularly within the African continent, given that legal systems in most African countries are particularly weak. It thus dissects the provisions that outline the principle of complementarity in tandem with the Court’s interpretation and application of complementarity in practice. Furthermore, through an exploratory survey of the referral of the Situation in Uganda, and the ICC Prosecutor’s proprio motu investigation of the Situation in Kenya, the thesis illustrates how a positive approach to complementarity can help establish a healthy cooperative synergy between the ICC and States, thereby promoting a functional expeditious criminal justice system. This will go a long way towards assuaging State’s fears that the ICC merely pays lip service to complementarity and arbitrarily supersedes national jurisdiction.
- Full Text:
The development of sustainability criteria to facilitate the selection of sanitation technologies within the Buffalo City Municipality (Eastern Cape Province, South Africa)
- Authors: Hoossein, Shafick
- Date: 2009
- Subjects: Sanitation -- South Africa -- Buffalo City Sustainable development -- South Africa -- Buffalo City Water -- Purification -- South Africa -- Buffalo City Sewage disposal -- South Africa -- Buffalo City Water quality -- South Africa -- Buffalo City Environmental health -- South Africa -- Buffalo City Environmental management -- South Africa -- Buffalo City
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:32275 , http://hdl.handle.net/10962/d1004595
- Description: Two and a half billion people mostly in sub-Saharan Africa and southern Asia remain without improved sanitation facilities despite the Millennium Development Goal 7's target to halve this number by 2015. While it might be tempting for developing countries such as South Africa to implement the cheapest and most rapidly constructed sanitation services simply to meet the desired target, this could have significant negative implications on human health and the environment over the long-term. As a result, there is a need to ensure that the most appropriate sustainable sanitation technologies are selected during the planning stage. The purpose of this research was therefore to document the development and pilot application of a flexible context-specific decision-support tool for sustainable sanitation technology selection within Buffalo City Municipality (BCM) in the Eastern Cape Province of South Africa. The first step in the development process was to ascertain the current status of sanitation within the municipality, with a specific focus on the main challenges related to the provision, maintenance and performance of these technologies. Thereafter, a participatory approach was employed involving BCM stakeholders to develop a series of sustainability criteria and indicators that took into consideration economic, social, environmental and technical concerns as well as legal requirements. The development process resulted in a list of 38 BCM sustainable sanitation selection criteria that were applied in a pilot study involving rural, urban and peri-urban communities within BCM. Certain criteria related to topographical features and the availability of piped water and land were considered useful for 'coarse screening' while others were applied during 'fine screening'. In order to enhance the context specificity of criteria, each was weighted, through consultation with key BCM stakeholders. This research confirmed that the sanitation situation in BCM was poor and preliminary evidence indicated that sanitation systems were negatively impacting on the quality of water resources within BCM justifying the need for a sustainable sanitation decision support tool. Based on the pilot application of the BCM Sustainability Selection Criteria List, the urine diversion technology was considered the most sustainable technology option in each study site. There were however, certain social criteria that received relatively low sustainability scores and these would need to be addressed prior to the approval of this technology for implementation. The approach adopted in this thesis was considered highly context-specific yet flexible and appropriate for adoption not only by BCM but other municipalities on a range of spatial scales.
- Full Text:
- Authors: Hoossein, Shafick
- Date: 2009
- Subjects: Sanitation -- South Africa -- Buffalo City Sustainable development -- South Africa -- Buffalo City Water -- Purification -- South Africa -- Buffalo City Sewage disposal -- South Africa -- Buffalo City Water quality -- South Africa -- Buffalo City Environmental health -- South Africa -- Buffalo City Environmental management -- South Africa -- Buffalo City
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:32275 , http://hdl.handle.net/10962/d1004595
- Description: Two and a half billion people mostly in sub-Saharan Africa and southern Asia remain without improved sanitation facilities despite the Millennium Development Goal 7's target to halve this number by 2015. While it might be tempting for developing countries such as South Africa to implement the cheapest and most rapidly constructed sanitation services simply to meet the desired target, this could have significant negative implications on human health and the environment over the long-term. As a result, there is a need to ensure that the most appropriate sustainable sanitation technologies are selected during the planning stage. The purpose of this research was therefore to document the development and pilot application of a flexible context-specific decision-support tool for sustainable sanitation technology selection within Buffalo City Municipality (BCM) in the Eastern Cape Province of South Africa. The first step in the development process was to ascertain the current status of sanitation within the municipality, with a specific focus on the main challenges related to the provision, maintenance and performance of these technologies. Thereafter, a participatory approach was employed involving BCM stakeholders to develop a series of sustainability criteria and indicators that took into consideration economic, social, environmental and technical concerns as well as legal requirements. The development process resulted in a list of 38 BCM sustainable sanitation selection criteria that were applied in a pilot study involving rural, urban and peri-urban communities within BCM. Certain criteria related to topographical features and the availability of piped water and land were considered useful for 'coarse screening' while others were applied during 'fine screening'. In order to enhance the context specificity of criteria, each was weighted, through consultation with key BCM stakeholders. This research confirmed that the sanitation situation in BCM was poor and preliminary evidence indicated that sanitation systems were negatively impacting on the quality of water resources within BCM justifying the need for a sustainable sanitation decision support tool. Based on the pilot application of the BCM Sustainability Selection Criteria List, the urine diversion technology was considered the most sustainable technology option in each study site. There were however, certain social criteria that received relatively low sustainability scores and these would need to be addressed prior to the approval of this technology for implementation. The approach adopted in this thesis was considered highly context-specific yet flexible and appropriate for adoption not only by BCM but other municipalities on a range of spatial scales.
- Full Text:
Purification, characterisation and application of inulinase and transferase enzymes in the production of fructose and oligosaccharides
- Authors: Mutanda, Taurai
- Date: 2008
- Subjects: Fructose Transferases Oligosaccharides
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4448 , http://hdl.handle.net/10962/d1007734
- Description: Inulin hydrolysis can occur as a result of the action of exoinulinases and endoinulinases acting alone or synergistically. Exoinulinases cleave the non-reducing β-(2, I) end of inulin releasing fructose while endoinulinases act on the internal linkages randomly to release inulotrioses (F₃), inulotetraoses (F₄) and inulopentaoses (F₅) as major products. Fructosyltransferases act by cleaving a sucrose molecule and then transferring the liberated fructose molecule to an acceptor molecule such as sucrose or another oligosaccharide to elongate the short chain fructooligosaccharide. The production of high yields of oligosaccharides of specific chain length from simple raw materials such as inulin and sucrose is a challenge. Oligosaccharides of chain length up to degree of polymerisation (DP) 5 and fructose were produced using preparations of three commercial microbial enzymes. Production of these novel oligosaccharides was achieved by employing response surface methodology (RSM) with central composite experimental design (CCD) for optimising product yield. Using a crude Novozyme 960 endoinulinase preparation isolated from Aspergillus niger, the following conditions gave a high inulooligosaccharide (lOS) yield, temperature (60 ºC), 150 g/L inulin concentration, 48 h incubation; pH 6.0 and enzyme dosage of 60 U/ml. Under these conditions, inulotrioses (70.3 mM), inulotetraoses (38.8 mM), and inulopentaoses, (3.5 mM) were produced. Response surface regression predicted similar product levels under similar conditions. The crude endoinulinase was purified through a three step purification procedure with a yield of 1.11 % and 3.5 fold purification. The molecular weight of this endoinulinase was estimated to be 68 .1 kDa by SDS-PAGE and its endoinulinase nature was confirmed by native PAGE. The purified endoinulinase was more efficient in production of lOS than the crude endoinulinase preparation. The purified endoinulinase demonstrated a high affinity for the inulin substrate (Km[subscript] 3.53 mM, Vmax[subscript] 666.67 μmol/min/ml). Pectinex Ultra SP-L, a commercial crude enzyme preparation isolated from Aspergillus aculeatus is a cocktail of several enzymes including a fructosyltransferase. The crude enzyme showed both transfructosylation and hydrolytic activity in 200 to 600 g/L sucrose. The main fructooligosaccharides produced from sucrose were l-kestose (GF₂), nystose (GF₃) and fructofuranosyl nystose (GF₄). After the first RSM, with the coded independent variables of temperature, incubation time, pH and sucrose concentration, the highest levels of GF₂, was 68.61 mM, under sucrose concentration 600 g/L, temperature 60°C, enzyme dosage 20 U/ml , pH 5.6, after 4 h incubation. A sucrose concentration of 400 g/L favoured the synthesis of high levels of GF₃ and GF₄. In the second RSM the maximal yields of GF₂, GF₃ and GF₄ were 152.07 mM, 131.38 mM and 43.99 mM respectively. A purified fructosyltransferase did not synthesise GF₄. Ammonium ions were demonstrated to enhance the yield of FOS. A mixture of glucose and fructose was used as substrate for FOS synthesis and no FOS were formed. Glucose was shown to be an end product inhibitor of the fructosyltransferase and therefore hinders the formation of high FOS yield. Fructozyme, isolated from Aspergillus ficuum is a mixture of exo and endoinulinases with the former being predominant was used for fructose production from inulin hydrolysis. The exoinulinase was purified to electrophoretic homogeneity by a three step purification procedure. The molecular weight of the enzyme was estimated to be 53 kDa with a 2 I % yield and 4.2-fold. Response surface regression was used to predict the maximum fructose levels achievable under the combinations of temperature, enzyme dosage and incubation time. A reaction time (48 h), enzyme dosage (100 U/ml) and inulin concentration (150 g/l) at pH 5.0 at 50°C gave higher fructose levels (106.6 mg/ml) using crude exoinulinase as compared to 98.43 mg/ml using the purified exoinulinase. These findings indicate that higher levels of fructose require longer incubation periods and higher inulin substrate concentrations with higher enzyme dosage. The crude exoinulinase preparation gave fairly higher levels of fructose than the purified exoinulinase and this is due to the presence of other hydrolytic enzymes in the crude preparation. The conditions established by RSM and CCO were adequate in producing high yield of oligosaccharides and fructose and can therefore be applied for their industrial production since they are in high demand due to their health benefits as prebiotics.
- Full Text:
- Authors: Mutanda, Taurai
- Date: 2008
- Subjects: Fructose Transferases Oligosaccharides
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4448 , http://hdl.handle.net/10962/d1007734
- Description: Inulin hydrolysis can occur as a result of the action of exoinulinases and endoinulinases acting alone or synergistically. Exoinulinases cleave the non-reducing β-(2, I) end of inulin releasing fructose while endoinulinases act on the internal linkages randomly to release inulotrioses (F₃), inulotetraoses (F₄) and inulopentaoses (F₅) as major products. Fructosyltransferases act by cleaving a sucrose molecule and then transferring the liberated fructose molecule to an acceptor molecule such as sucrose or another oligosaccharide to elongate the short chain fructooligosaccharide. The production of high yields of oligosaccharides of specific chain length from simple raw materials such as inulin and sucrose is a challenge. Oligosaccharides of chain length up to degree of polymerisation (DP) 5 and fructose were produced using preparations of three commercial microbial enzymes. Production of these novel oligosaccharides was achieved by employing response surface methodology (RSM) with central composite experimental design (CCD) for optimising product yield. Using a crude Novozyme 960 endoinulinase preparation isolated from Aspergillus niger, the following conditions gave a high inulooligosaccharide (lOS) yield, temperature (60 ºC), 150 g/L inulin concentration, 48 h incubation; pH 6.0 and enzyme dosage of 60 U/ml. Under these conditions, inulotrioses (70.3 mM), inulotetraoses (38.8 mM), and inulopentaoses, (3.5 mM) were produced. Response surface regression predicted similar product levels under similar conditions. The crude endoinulinase was purified through a three step purification procedure with a yield of 1.11 % and 3.5 fold purification. The molecular weight of this endoinulinase was estimated to be 68 .1 kDa by SDS-PAGE and its endoinulinase nature was confirmed by native PAGE. The purified endoinulinase was more efficient in production of lOS than the crude endoinulinase preparation. The purified endoinulinase demonstrated a high affinity for the inulin substrate (Km[subscript] 3.53 mM, Vmax[subscript] 666.67 μmol/min/ml). Pectinex Ultra SP-L, a commercial crude enzyme preparation isolated from Aspergillus aculeatus is a cocktail of several enzymes including a fructosyltransferase. The crude enzyme showed both transfructosylation and hydrolytic activity in 200 to 600 g/L sucrose. The main fructooligosaccharides produced from sucrose were l-kestose (GF₂), nystose (GF₃) and fructofuranosyl nystose (GF₄). After the first RSM, with the coded independent variables of temperature, incubation time, pH and sucrose concentration, the highest levels of GF₂, was 68.61 mM, under sucrose concentration 600 g/L, temperature 60°C, enzyme dosage 20 U/ml , pH 5.6, after 4 h incubation. A sucrose concentration of 400 g/L favoured the synthesis of high levels of GF₃ and GF₄. In the second RSM the maximal yields of GF₂, GF₃ and GF₄ were 152.07 mM, 131.38 mM and 43.99 mM respectively. A purified fructosyltransferase did not synthesise GF₄. Ammonium ions were demonstrated to enhance the yield of FOS. A mixture of glucose and fructose was used as substrate for FOS synthesis and no FOS were formed. Glucose was shown to be an end product inhibitor of the fructosyltransferase and therefore hinders the formation of high FOS yield. Fructozyme, isolated from Aspergillus ficuum is a mixture of exo and endoinulinases with the former being predominant was used for fructose production from inulin hydrolysis. The exoinulinase was purified to electrophoretic homogeneity by a three step purification procedure. The molecular weight of the enzyme was estimated to be 53 kDa with a 2 I % yield and 4.2-fold. Response surface regression was used to predict the maximum fructose levels achievable under the combinations of temperature, enzyme dosage and incubation time. A reaction time (48 h), enzyme dosage (100 U/ml) and inulin concentration (150 g/l) at pH 5.0 at 50°C gave higher fructose levels (106.6 mg/ml) using crude exoinulinase as compared to 98.43 mg/ml using the purified exoinulinase. These findings indicate that higher levels of fructose require longer incubation periods and higher inulin substrate concentrations with higher enzyme dosage. The crude exoinulinase preparation gave fairly higher levels of fructose than the purified exoinulinase and this is due to the presence of other hydrolytic enzymes in the crude preparation. The conditions established by RSM and CCO were adequate in producing high yield of oligosaccharides and fructose and can therefore be applied for their industrial production since they are in high demand due to their health benefits as prebiotics.
- Full Text:
The toxic effect of heavy metals on algal biomass (Spirulina sp.) and carbonic anhydrase activity, an enzyme which is central to algal application in metal precipitation
- Authors: Nightingale, Leigh
- Date: 2004
- Subjects: Heavy metals -- Toxicology , Spirulina , Carbonic anhydrase , Algae -- Metabolism , Photosynthesis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4093 , http://hdl.handle.net/10962/d1007858 , Heavy metals -- Toxicology , Spirulina , Carbonic anhydrase , Algae -- Metabolism , Photosynthesis
- Description: Acid rmne drainage (AMD) is a major pollution problem througbout the world, adversely affecting both surface and groundwaters. AMD is principally associated with the mining of sulphide ores. The most commonly associated minerals being sulphur, copper, zinc, silver, gold, lead and uranium. As conventional methods for removing heavy metals from wastewater are often prohibitively expensive, the implementation of biological processes for the removal of heavy metals has become a realistic practice. The objectives of this project was firstly to establish the effect of copper, lead and nickel, heavy metals commonly found in AMD waters, on the enzyme carbonic anhydrase, which is an integral part of the carbon concentrating mechanism (CCM) and secondly, to determine the feasibility of using the alkalinity generated by Spindina for the precipitation of heavy metals from solution. Initially, batch flask experiments were performed and it was found that the algae were able to utilise the bicarbonate supplied in the medium, under CO, limiting conditions, through the induction of their CCM, resulting in the generation of carbonate. The effect of the inhibitors, acetazolamide (AZ) and ethoxyzolamide (EZ), were also investigated in order to determine the importance of carbonic anhydrase (CA) in inorganic carbon accumulation and photosynthesis. Results obtained were consistent with those observed in literature and it was found that at IOOf.LM AZ and EZ, complete inhibition of photosynthesis and carbonic anhydrase occurred, with no oxygen being evolved. The results obtained from the inhibitor experiments substantiate the findings that carbonic anhydrase is an important part of the CCM, and that the dehydration of bicarbonate to carbon dioxide and hydroxide ions, is in fact an enzymatic process regulated by the enzyme carbonic anhydrase and is essential for efficient photosynthesis. The effect of heavy metals on Spirulina was also investigated. Lead, copper and nickel were all found to cause a reduction in the synthesis of chlorophyll a, which resulted in a decrease in photosynthetic efficiency and eventually death of the culture. The morphology of the algae was also severely affected by heavy metals, with degradation and aJmost complete disintegration of the algal filaments occurring. Using the Wilbur-Anderson assay method, carbonic anhydrase activity was found to be lower in the experimental flasks containing heavy metals, than the control flasks, reducing the algae's ability to utilise the bicarbonate in solution for effective photosynthesis. The Wilbur-Anderson assay method did not prove to be a reliable method for measuring changes in enzyme activity as results were found to be erratic. Therefore attempts were made to use an oxygen electrode as an alternative method for determining the effects of various parameters on enzyme activity and photosynthesis, this proved to be more successful. Because of the toxic effects of heavy metals on Spirulina it was decided that the use of the biogenic alkalinity generated by the algae for the precipitation of heavy metals may be successfully employed as an alternative method for bioremediation and metal recovery. Carbonate reacts readily with metals, therefore the carbonate produced by this algal system was used for the precipitation of metals. It was possible to categorise the precipitation reactions observed into three groups, namely those metals which, a) precipitate as hydroxides, b) precipitate as carbonates generated from the dissociation of bicarbonate and c) metals which can only precipitate if there is free carbonate present in solution.
- Full Text:
- Authors: Nightingale, Leigh
- Date: 2004
- Subjects: Heavy metals -- Toxicology , Spirulina , Carbonic anhydrase , Algae -- Metabolism , Photosynthesis
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4093 , http://hdl.handle.net/10962/d1007858 , Heavy metals -- Toxicology , Spirulina , Carbonic anhydrase , Algae -- Metabolism , Photosynthesis
- Description: Acid rmne drainage (AMD) is a major pollution problem througbout the world, adversely affecting both surface and groundwaters. AMD is principally associated with the mining of sulphide ores. The most commonly associated minerals being sulphur, copper, zinc, silver, gold, lead and uranium. As conventional methods for removing heavy metals from wastewater are often prohibitively expensive, the implementation of biological processes for the removal of heavy metals has become a realistic practice. The objectives of this project was firstly to establish the effect of copper, lead and nickel, heavy metals commonly found in AMD waters, on the enzyme carbonic anhydrase, which is an integral part of the carbon concentrating mechanism (CCM) and secondly, to determine the feasibility of using the alkalinity generated by Spindina for the precipitation of heavy metals from solution. Initially, batch flask experiments were performed and it was found that the algae were able to utilise the bicarbonate supplied in the medium, under CO, limiting conditions, through the induction of their CCM, resulting in the generation of carbonate. The effect of the inhibitors, acetazolamide (AZ) and ethoxyzolamide (EZ), were also investigated in order to determine the importance of carbonic anhydrase (CA) in inorganic carbon accumulation and photosynthesis. Results obtained were consistent with those observed in literature and it was found that at IOOf.LM AZ and EZ, complete inhibition of photosynthesis and carbonic anhydrase occurred, with no oxygen being evolved. The results obtained from the inhibitor experiments substantiate the findings that carbonic anhydrase is an important part of the CCM, and that the dehydration of bicarbonate to carbon dioxide and hydroxide ions, is in fact an enzymatic process regulated by the enzyme carbonic anhydrase and is essential for efficient photosynthesis. The effect of heavy metals on Spirulina was also investigated. Lead, copper and nickel were all found to cause a reduction in the synthesis of chlorophyll a, which resulted in a decrease in photosynthetic efficiency and eventually death of the culture. The morphology of the algae was also severely affected by heavy metals, with degradation and aJmost complete disintegration of the algal filaments occurring. Using the Wilbur-Anderson assay method, carbonic anhydrase activity was found to be lower in the experimental flasks containing heavy metals, than the control flasks, reducing the algae's ability to utilise the bicarbonate in solution for effective photosynthesis. The Wilbur-Anderson assay method did not prove to be a reliable method for measuring changes in enzyme activity as results were found to be erratic. Therefore attempts were made to use an oxygen electrode as an alternative method for determining the effects of various parameters on enzyme activity and photosynthesis, this proved to be more successful. Because of the toxic effects of heavy metals on Spirulina it was decided that the use of the biogenic alkalinity generated by the algae for the precipitation of heavy metals may be successfully employed as an alternative method for bioremediation and metal recovery. Carbonate reacts readily with metals, therefore the carbonate produced by this algal system was used for the precipitation of metals. It was possible to categorise the precipitation reactions observed into three groups, namely those metals which, a) precipitate as hydroxides, b) precipitate as carbonates generated from the dissociation of bicarbonate and c) metals which can only precipitate if there is free carbonate present in solution.
- Full Text:
Valuation of intellectual capital in South African companies: a comparative study of three valuation methods
- Authors: Maree, Kevin W
- Date: 2002
- Subjects: Accounting
- Language: English
- Type: Thesis , Masters , MCom
- Identifier: vital:877 , http://hdl.handle.net/10962/d1001631
- Description: This study discusses three valuation methods for intellectual capital and considers two of these (Tobin’s “q” and CIV) as suitable valuation methods.
- Full Text:
- Authors: Maree, Kevin W
- Date: 2002
- Subjects: Accounting
- Language: English
- Type: Thesis , Masters , MCom
- Identifier: vital:877 , http://hdl.handle.net/10962/d1001631
- Description: This study discusses three valuation methods for intellectual capital and considers two of these (Tobin’s “q” and CIV) as suitable valuation methods.
- Full Text:
An examination of the facilitatory role for environmental education of conservancies
- Authors: Law-Jackson, Danielle
- Date: 2000
- Subjects: Environmental education Nature conservation -- South Africa Wildlife conservation -- South Africa Conservation of natural resources -- South Africa
- Language: English
- Type: Thesis , Masters , MEd
- Identifier: vital:1529 , http://hdl.handle.net/10962/d1003411
- Description: This study examines the possibilities within the conservancy movement for the facilitation of environmental education. By means of a case study approach, a conservancy in the Northern Cape and the conservancy movement within the Free State provinces are compared with a view to elucidating factors militating against and promoting the success of environmental education programmes. The current lack of research in isolated rural areas is discussed. Re-orientation of perspectives regarding the value of conservancy-based environmental education within governmental, academic and local circles is recommended.
- Full Text:
- Authors: Law-Jackson, Danielle
- Date: 2000
- Subjects: Environmental education Nature conservation -- South Africa Wildlife conservation -- South Africa Conservation of natural resources -- South Africa
- Language: English
- Type: Thesis , Masters , MEd
- Identifier: vital:1529 , http://hdl.handle.net/10962/d1003411
- Description: This study examines the possibilities within the conservancy movement for the facilitation of environmental education. By means of a case study approach, a conservancy in the Northern Cape and the conservancy movement within the Free State provinces are compared with a view to elucidating factors militating against and promoting the success of environmental education programmes. The current lack of research in isolated rural areas is discussed. Re-orientation of perspectives regarding the value of conservancy-based environmental education within governmental, academic and local circles is recommended.
- Full Text:
The psychological adjustment of middle managers after revolutionary organisational change
- Authors: Coates, Nicholas Robert
- Date: 1999
- Subjects: Psychology, Industrial -- South Africa Organizational change -- South Africa Organizational change -- Psychological aspects Management -- Psychological aspects
- Language: English
- Type: Thesis , Masters , MSocSc
- Identifier: vital:3190 , http://hdl.handle.net/10962/d1008554
- Description:
With the accelerated process of political and socio-economic transformation in South Africa, revolutionary organisational change has become a given in contemporary South African business life (Human & Horwitz, 1992). For revolutionary organisational change to succeed in South Africa, middle managers who represent the 'cement' of the organisation, need to adjust at the individual level. However, the literature on organisational change remains curiously silent about individual adjustment (Ashford, 1988). The goals of the research were firstly, to recount the middle manager's perceptions and experiences of revolutionary organisational change. Secondly, to detail the psychological re-
- Full Text:
- Authors: Coates, Nicholas Robert
- Date: 1999
- Subjects: Psychology, Industrial -- South Africa Organizational change -- South Africa Organizational change -- Psychological aspects Management -- Psychological aspects
- Language: English
- Type: Thesis , Masters , MSocSc
- Identifier: vital:3190 , http://hdl.handle.net/10962/d1008554
- Description:
With the accelerated process of political and socio-economic transformation in South Africa, revolutionary organisational change has become a given in contemporary South African business life (Human & Horwitz, 1992). For revolutionary organisational change to succeed in South Africa, middle managers who represent the 'cement' of the organisation, need to adjust at the individual level. However, the literature on organisational change remains curiously silent about individual adjustment (Ashford, 1988). The goals of the research were firstly, to recount the middle manager's perceptions and experiences of revolutionary organisational change. Secondly, to detail the psychological re-
- Full Text:
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