Profiling for volatile compounds in the Kgalagadi desert truffle
- Authors: Bogopa, Juda
- Date: 2013
- Subjects: Desert plants -- Botswana -- Kgalagadi District Supercritical fluid extraction Gas chromatography Mass spectrometry Tuberaceae -- Analysis -- Botswana -- Kgalagadi District Truffles -- Analysis -- Botswana -- Kgalagadi District
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4280 , http://hdl.handle.net/10962/d1002006
- Description: The thesis focuses on ‘green’ sample preparation techniques that utilise minimal or no organic solvents thereby producing negligible volumes of organic waste, to ensure safety both to user and the environment. Volatile compounds were extracted and pre-concentrated from Kgalagadi desert truffles (kalaharituber) by headspace solid phase microextraction (HSSPME) and supercritical fluid extraction (SFE). PHWE was employed for the extraction of amino acids and fatty acids. Subsequent analysis of volatile compounds was carried out by gas chromatography coupled with mass spectrometry. Four types of HS-SPME fibers (PDMS 100 μm, PDMS 7 μm, Polyacrylate 85 μm, CAR/DVB/PDMS 50/30 μm) were evaluated. A total of 24 volatile compounds with a molecular weight range from 110 to 354, the most prominent peak being 2-t-Butyl-2,3-dimethyl-3-buten-1-ol (C₁₀H₂₀O, MW 156) were detected after sampling with a PDMS 100 μm fiber. Less volatile compounds were detected after SFE with CO2. A total of 16 amino acids were identified while 17 fatty acids (MW from 132.12 to 367.49) were also identified. The characteristic profile of the Kgalagadi desert truffle was found to contain mainly fatty acid methyl esters and unsaturated aliphatic hydrocarbons. The most prominent compound peaks identified were; 2-t-butyl-2,3-dimethyl- 3-buten-1-ol, disulfide, ethyl benzoic acid 2-4-dhydroxy-3,6-dimethyl-methyl ester, 8,11- octadecanoic acid methyl ester, benzoic acid, 2,4-dihydroxy-3,6-dimethyl-methyl ester, isoquinoline, 1 butyl-3,4-dihydro and 3-heptanone, 6 methyl. Optimization results indicated that fresh slices from the heart of truffles were the best to use for HS-SPME-GCMS volatile compound analysis as they showed a higher sensitivity
- Full Text:
- Date Issued: 2013
- Authors: Bogopa, Juda
- Date: 2013
- Subjects: Desert plants -- Botswana -- Kgalagadi District Supercritical fluid extraction Gas chromatography Mass spectrometry Tuberaceae -- Analysis -- Botswana -- Kgalagadi District Truffles -- Analysis -- Botswana -- Kgalagadi District
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4280 , http://hdl.handle.net/10962/d1002006
- Description: The thesis focuses on ‘green’ sample preparation techniques that utilise minimal or no organic solvents thereby producing negligible volumes of organic waste, to ensure safety both to user and the environment. Volatile compounds were extracted and pre-concentrated from Kgalagadi desert truffles (kalaharituber) by headspace solid phase microextraction (HSSPME) and supercritical fluid extraction (SFE). PHWE was employed for the extraction of amino acids and fatty acids. Subsequent analysis of volatile compounds was carried out by gas chromatography coupled with mass spectrometry. Four types of HS-SPME fibers (PDMS 100 μm, PDMS 7 μm, Polyacrylate 85 μm, CAR/DVB/PDMS 50/30 μm) were evaluated. A total of 24 volatile compounds with a molecular weight range from 110 to 354, the most prominent peak being 2-t-Butyl-2,3-dimethyl-3-buten-1-ol (C₁₀H₂₀O, MW 156) were detected after sampling with a PDMS 100 μm fiber. Less volatile compounds were detected after SFE with CO2. A total of 16 amino acids were identified while 17 fatty acids (MW from 132.12 to 367.49) were also identified. The characteristic profile of the Kgalagadi desert truffle was found to contain mainly fatty acid methyl esters and unsaturated aliphatic hydrocarbons. The most prominent compound peaks identified were; 2-t-butyl-2,3-dimethyl- 3-buten-1-ol, disulfide, ethyl benzoic acid 2-4-dhydroxy-3,6-dimethyl-methyl ester, 8,11- octadecanoic acid methyl ester, benzoic acid, 2,4-dihydroxy-3,6-dimethyl-methyl ester, isoquinoline, 1 butyl-3,4-dihydro and 3-heptanone, 6 methyl. Optimization results indicated that fresh slices from the heart of truffles were the best to use for HS-SPME-GCMS volatile compound analysis as they showed a higher sensitivity
- Full Text:
- Date Issued: 2013
The effectiveness of livestock guarding dogs for livestock production and conservation in Namibia
- Authors: Potgieter, Gail Christine
- Date: 2011
- Subjects: Livestock protection dogs -- Namibia , Herding dogs -- Namibia , Livestock -- Predators of -- Control -- Namibia , Livestock -- Losses -- Namibia
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10680 , http://hdl.handle.net/10948/1666 , Livestock protection dogs -- Namibia , Herding dogs -- Namibia , Livestock -- Predators of -- Control -- Namibia , Livestock -- Losses -- Namibia
- Description: The use of livestock guarding dogs (LGDs) to mitigate farmer-predator conflict in Namibia was evaluated. As farmer-predator conflict has two sides, LGDs were evaluated in terms of livestock production and conservation. The main objectives in terms of livestock production were to document: 1) the perceived ability of LGDs to reduce livestock losses in a cost-effective manner; 2) the farmers’ satisfaction with LGD performance; and 3) factors influencing LGD behaviour. The main objectives in terms of conservation were to record: 1) predator killing by farmers relative to LGD introduction; 2) direct impacts of LGDs on target (damage-causing) species; and 3) the impact of LGDs on non-target species. This evaluation was conducted on LGDs bred by the Cheetah Conservation Fund (CCF) and placed on farms in Namibia. The data were collected during face-to-face interviews with farmers using LGDs. Historical data from the CCF programme were used in conjunction with a complete survey of the farmers in the CCF LGD programme during 2009-2010. In terms of livestock production, 91 percent of the LGDs (n = 65) eliminated or reduced livestock losses. Subsequently, 73 percent of the farmers perceived their LGDs as economically beneficial, although a cost-benefit analysis showed that only 59 percent of the LGDs were cost-effective. Farmers were generally satisfied with the performance of their LGDs. However, farmer satisfaction was more closely linked to good LGD behaviour than the perceived reduction in livestock losses. The most commonly-reported LGD behavioural problems (n = 195) were staying at home rather than accompanying the livestock (21 percent) and chasing wildlife (19 percent). LGD staying home behaviour was linked to a lack of care on subsistence farms, as high quality dog food was not consistently provided. Care for LGDs declined with LGD age on subsistence, but not commercial, farms. In terms of conservation, predator-killing farmers killed fewer individuals in the year since LGD introduction than previously; this result was only significant for black-backed jackal Canis mesomelas. However, 37 LGDs killed jackals, nine killed baboons Papio ursinus, three killed caracals Caracal caracal and one killed a cheetah Acinonyx jubatus (n = 83). Farmers and LGDs combined killed significantly more jackals in the survey year than the same farmers (n = 36) killed before LGD introduction. Conversely, five farmers killed 3.2 ± 2.01 cheetahs each in the year before LGD introduction, whereas LGDs and these farmers combined killed only 0.2 ± 0.2 cheetahs per farm in the survey year. Only 16 LGDs (n = 83) killed non-target species. The high LGD success rate in terms of livestock production was facilitated by livestock husbandry practices in the study area. In terms of conservation, LGDs were more beneficial for apex predators than for mesopredators and had a minor impact on non-target species.
- Full Text:
- Date Issued: 2011
- Authors: Potgieter, Gail Christine
- Date: 2011
- Subjects: Livestock protection dogs -- Namibia , Herding dogs -- Namibia , Livestock -- Predators of -- Control -- Namibia , Livestock -- Losses -- Namibia
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:10680 , http://hdl.handle.net/10948/1666 , Livestock protection dogs -- Namibia , Herding dogs -- Namibia , Livestock -- Predators of -- Control -- Namibia , Livestock -- Losses -- Namibia
- Description: The use of livestock guarding dogs (LGDs) to mitigate farmer-predator conflict in Namibia was evaluated. As farmer-predator conflict has two sides, LGDs were evaluated in terms of livestock production and conservation. The main objectives in terms of livestock production were to document: 1) the perceived ability of LGDs to reduce livestock losses in a cost-effective manner; 2) the farmers’ satisfaction with LGD performance; and 3) factors influencing LGD behaviour. The main objectives in terms of conservation were to record: 1) predator killing by farmers relative to LGD introduction; 2) direct impacts of LGDs on target (damage-causing) species; and 3) the impact of LGDs on non-target species. This evaluation was conducted on LGDs bred by the Cheetah Conservation Fund (CCF) and placed on farms in Namibia. The data were collected during face-to-face interviews with farmers using LGDs. Historical data from the CCF programme were used in conjunction with a complete survey of the farmers in the CCF LGD programme during 2009-2010. In terms of livestock production, 91 percent of the LGDs (n = 65) eliminated or reduced livestock losses. Subsequently, 73 percent of the farmers perceived their LGDs as economically beneficial, although a cost-benefit analysis showed that only 59 percent of the LGDs were cost-effective. Farmers were generally satisfied with the performance of their LGDs. However, farmer satisfaction was more closely linked to good LGD behaviour than the perceived reduction in livestock losses. The most commonly-reported LGD behavioural problems (n = 195) were staying at home rather than accompanying the livestock (21 percent) and chasing wildlife (19 percent). LGD staying home behaviour was linked to a lack of care on subsistence farms, as high quality dog food was not consistently provided. Care for LGDs declined with LGD age on subsistence, but not commercial, farms. In terms of conservation, predator-killing farmers killed fewer individuals in the year since LGD introduction than previously; this result was only significant for black-backed jackal Canis mesomelas. However, 37 LGDs killed jackals, nine killed baboons Papio ursinus, three killed caracals Caracal caracal and one killed a cheetah Acinonyx jubatus (n = 83). Farmers and LGDs combined killed significantly more jackals in the survey year than the same farmers (n = 36) killed before LGD introduction. Conversely, five farmers killed 3.2 ± 2.01 cheetahs each in the year before LGD introduction, whereas LGDs and these farmers combined killed only 0.2 ± 0.2 cheetahs per farm in the survey year. Only 16 LGDs (n = 83) killed non-target species. The high LGD success rate in terms of livestock production was facilitated by livestock husbandry practices in the study area. In terms of conservation, LGDs were more beneficial for apex predators than for mesopredators and had a minor impact on non-target species.
- Full Text:
- Date Issued: 2011
Solid waste management and its implications on livestock health in selected abattoirs of the Eastern Cape province, South Africa
- Authors: Nongcula, Vikhaya Vincent
- Date: 2016
- Subjects: Refuse and refuse disposal -- South Africa -- Eastern Cape Recycling (Waste, etc.) -- South Africa -- Eastern Cape Livestock -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/12293 , vital:39250
- Description: An investigation was carried out on the implications of solid waste management on livestock health in selected abattoirs of the Eastern Cape Province of South Africa. The main objective of the study was to determine the effects of solid waste management on livestock health as well as to assess the perceptions of solid waste management among livestock owners. In this study, a questionnaire was designed to obtain information regarding farmer’s perceptions on solid waste and how it affects animal health, and active abattoir inspection were conducted to assess how solid waste affects the animal. The results revealed that most farmers practiced poor solid waste management as a result animals were exposed to ingestion of indigestible materials. This was confirmed on the basis that most farmers (99.2 percent) had no municipal dustbins in their areas, no recycling programmes (94.2 percent) and 51.7percent of the farmers disposed their litter in open land as a result most farmers (75.9percent) agreed that solid waste management was a problem in their areas. Due to poor solid waste management, 53.8 percent of the farmers confirmed that they have seen livestock ingesting indigestible materials such as plastics and 69.0 percent of the farmers suspected that ingestion of such materials caused some of the health issue experienced by animals on their farm. The active abattoir inspection revealed that plastics, wires, nails, ropes, stones, bones, cloths, hairs and polybezoars were frequently occurring in slaughter cattle stomachs particularly in the rumen and reticulum and in female and older cattle. As a result most affected cattle were in thin and average condition. The effect of indigestible objects on body condition score (BCS) and organ condemnation was determined using linear regression analysis test; where BCS and condemnation were the dependent variables and indigestible objects were the independent variables. The results revealed that some organs were condemned in affected cattle and the statistical analysis showed that there was no significant difference (p< 0.05) between the prevalence of indigestible materials and organ condemnation. However, there was a significant difference between the prevalence of indigestible materials and body condition score of affected cattle although they were coming from different farming systems. It was concluded that livestock health was affected by solid waste and recommended that practice of good animal husbandry and proper solid waste management be practiced to overcome the implications (poor body condition score) of solid waste in livestock. Furthermore, close cooperation between government extension officers, commercial and communal farmers in knowledge-sharing regarding health issues of livestock is recommended.
- Full Text:
- Date Issued: 2016
- Authors: Nongcula, Vikhaya Vincent
- Date: 2016
- Subjects: Refuse and refuse disposal -- South Africa -- Eastern Cape Recycling (Waste, etc.) -- South Africa -- Eastern Cape Livestock -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/12293 , vital:39250
- Description: An investigation was carried out on the implications of solid waste management on livestock health in selected abattoirs of the Eastern Cape Province of South Africa. The main objective of the study was to determine the effects of solid waste management on livestock health as well as to assess the perceptions of solid waste management among livestock owners. In this study, a questionnaire was designed to obtain information regarding farmer’s perceptions on solid waste and how it affects animal health, and active abattoir inspection were conducted to assess how solid waste affects the animal. The results revealed that most farmers practiced poor solid waste management as a result animals were exposed to ingestion of indigestible materials. This was confirmed on the basis that most farmers (99.2 percent) had no municipal dustbins in their areas, no recycling programmes (94.2 percent) and 51.7percent of the farmers disposed their litter in open land as a result most farmers (75.9percent) agreed that solid waste management was a problem in their areas. Due to poor solid waste management, 53.8 percent of the farmers confirmed that they have seen livestock ingesting indigestible materials such as plastics and 69.0 percent of the farmers suspected that ingestion of such materials caused some of the health issue experienced by animals on their farm. The active abattoir inspection revealed that plastics, wires, nails, ropes, stones, bones, cloths, hairs and polybezoars were frequently occurring in slaughter cattle stomachs particularly in the rumen and reticulum and in female and older cattle. As a result most affected cattle were in thin and average condition. The effect of indigestible objects on body condition score (BCS) and organ condemnation was determined using linear regression analysis test; where BCS and condemnation were the dependent variables and indigestible objects were the independent variables. The results revealed that some organs were condemned in affected cattle and the statistical analysis showed that there was no significant difference (p< 0.05) between the prevalence of indigestible materials and organ condemnation. However, there was a significant difference between the prevalence of indigestible materials and body condition score of affected cattle although they were coming from different farming systems. It was concluded that livestock health was affected by solid waste and recommended that practice of good animal husbandry and proper solid waste management be practiced to overcome the implications (poor body condition score) of solid waste in livestock. Furthermore, close cooperation between government extension officers, commercial and communal farmers in knowledge-sharing regarding health issues of livestock is recommended.
- Full Text:
- Date Issued: 2016
In-beam spectroscopy of ⁷²Ge
- Authors: Henninger, Katharine Rose
- Date: 2012
- Subjects: Nuclear shapes , Hyperfine structure
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5559 , http://hdl.handle.net/10962/d1018233
- Description: The high-spin states of the nucleus ⁷²Ge were populated via the ⁷°Zn(a,2n)⁷²Ge fusion-evaporation reaction at a beam-energy of 30 MeV. The ')'-decays were observed with the AFRODITE spectrometer array at iThemba LABS (Faure, South Africa). Twenty-seven new transitions, 18 new levels and 2 new bands were added to the ⁷²Ge level scheme, and several corrections were made to t he previously-known level scheme. Transitions were characterised using Directional Correlation from Oriented states (DCO) and polarisation measurements, and spin-parity assignments were made for 7 new levels. Spin and parity were also reassigned to a previously-known structure. One of the new bands was ident ified as part of an octupole-vibrational band. No tetrahedral states were observed in the scheme despite N = 40 and Z = 32 having been identified as tetrahedral magic numbers. The meaning of the new 14⁺ yrast level for the underlying structure of the ground state band is discussed
- Full Text:
- Date Issued: 2012
- Authors: Henninger, Katharine Rose
- Date: 2012
- Subjects: Nuclear shapes , Hyperfine structure
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5559 , http://hdl.handle.net/10962/d1018233
- Description: The high-spin states of the nucleus ⁷²Ge were populated via the ⁷°Zn(a,2n)⁷²Ge fusion-evaporation reaction at a beam-energy of 30 MeV. The ')'-decays were observed with the AFRODITE spectrometer array at iThemba LABS (Faure, South Africa). Twenty-seven new transitions, 18 new levels and 2 new bands were added to the ⁷²Ge level scheme, and several corrections were made to t he previously-known level scheme. Transitions were characterised using Directional Correlation from Oriented states (DCO) and polarisation measurements, and spin-parity assignments were made for 7 new levels. Spin and parity were also reassigned to a previously-known structure. One of the new bands was ident ified as part of an octupole-vibrational band. No tetrahedral states were observed in the scheme despite N = 40 and Z = 32 having been identified as tetrahedral magic numbers. The meaning of the new 14⁺ yrast level for the underlying structure of the ground state band is discussed
- Full Text:
- Date Issued: 2012
Isolation, purification and characterization of inulin and fructooligosaccharides from chicorium intybus and inulinase from aspergillus niger
- Mavumengwana, Vuyo Bhongelethu
- Authors: Mavumengwana, Vuyo Bhongelethu
- Date: 2005
- Subjects: Aspergillus , Inulin , Chicory -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3954 , http://hdl.handle.net/10962/d1004013 , Aspergillus , Inulin , Chicory -- South Africa
- Description: Inulin is a non-digestible carbohydrate fructan polymer consisting mainly of β (1→2) fructosyl fructose links. Enzymatic hydrolysis of inulin by inulinase results in the production of low D.P (degree of polymerization) oligosaccharides also called fructooligosaccharides(FOS). Isolation of inulin from chicory root (Chicorium intybus) was achieved by first, extraction using deionized water (600C), followed by carbonation (0.1 M Ca(OH)2 and CO2 gas). This was filtered in order to remove the non sugars, thereafter, treated successfully with polyamide 6 powder. A cation exchanger and an anion exchanger were used to further exclude other components such as tannins and pigments. The extracted inulin was quantified using the Somogyi-Nelson colourimetric assay. Chicory root (207 g, 30 % being water) yielded 30 g of the raw extract. A 100 mg of the raw extract was assayed and found to contain 11 % yield of inulin which was 80.2 % in purity and 4 % free fructose. Analysis of the crude and purified inulin extracts on the MALDI TOF spectrometry showed the samples to have a DP of 2 to 22 and 2 to 27 respectively. Maximum inulinase production from Aspergillus niger grown on inulin was observed after 60 hours. The enzyme activity was found to be 1.168 U/ml with a temperature and pH optimum of 30 °C and 7.7 respectively. The enzyme proved to be unstable as it progressively lost its total activity during attempts at purification.
- Full Text:
- Date Issued: 2005
- Authors: Mavumengwana, Vuyo Bhongelethu
- Date: 2005
- Subjects: Aspergillus , Inulin , Chicory -- South Africa
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3954 , http://hdl.handle.net/10962/d1004013 , Aspergillus , Inulin , Chicory -- South Africa
- Description: Inulin is a non-digestible carbohydrate fructan polymer consisting mainly of β (1→2) fructosyl fructose links. Enzymatic hydrolysis of inulin by inulinase results in the production of low D.P (degree of polymerization) oligosaccharides also called fructooligosaccharides(FOS). Isolation of inulin from chicory root (Chicorium intybus) was achieved by first, extraction using deionized water (600C), followed by carbonation (0.1 M Ca(OH)2 and CO2 gas). This was filtered in order to remove the non sugars, thereafter, treated successfully with polyamide 6 powder. A cation exchanger and an anion exchanger were used to further exclude other components such as tannins and pigments. The extracted inulin was quantified using the Somogyi-Nelson colourimetric assay. Chicory root (207 g, 30 % being water) yielded 30 g of the raw extract. A 100 mg of the raw extract was assayed and found to contain 11 % yield of inulin which was 80.2 % in purity and 4 % free fructose. Analysis of the crude and purified inulin extracts on the MALDI TOF spectrometry showed the samples to have a DP of 2 to 22 and 2 to 27 respectively. Maximum inulinase production from Aspergillus niger grown on inulin was observed after 60 hours. The enzyme activity was found to be 1.168 U/ml with a temperature and pH optimum of 30 °C and 7.7 respectively. The enzyme proved to be unstable as it progressively lost its total activity during attempts at purification.
- Full Text:
- Date Issued: 2005
The impact of rangeland degradation on vegetation cover and soil quality in semi-arid communal rangelands of the Eastern Cape province, South Africa
- Authors: Maziko, Yonela
- Date: 2017
- Subjects: Communal rangelands -- South Africa -- Eastern Cape Land degradation -- South Africa -- Eastern Cape Vegetation management -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/15056 , vital:40159
- Description: This study was conducted to investigate the influence of degradation on vegetation and soil quality in two semi-arid rangelands in the Eastern Cape Province of South Africa. The study was undertaken in Kwezana and Esigingqini communal areas that showed signs id degradation. The assessment incorporated the soil and grass layer in different degradation levels, namely nondegraded, slightly degraded, moderately and severely degraded. The non-degraded has a sufficient amount of vegetation cover and species composition. The slightly degraded site consists of a few visible bare patches. The moderately degraded areas showed signs of active sheet erosion such as pedestals and a higher percentage of bare areas and the severely degraded area is characterized by dongas and gullies, with vast areas that have no ground cover or top fertile soil layer. Using a step point method, herbaceous species composition was determined within three transects from three plots 100 m x 40 m in each degradation level. The aboveground phytomass production was determined by harvesting herbaceous species within four randomly placed 0.25 m2 quadrats in each transect. Bare ground was determined by counting and recording the distance between tufts within each quadrat. Litter cover was determined by visual observation and rated on a scale of 1 to 5. A number of physical and chemical soil properties were used to quantify the effects of degradation on soil quality. To determine soil chemical properties, three soil samples were collected at a depth of 20 cm, in each plot and analyzed for pH, soil macro (N, C, K, P, Ca, Mg) and micro (Zn, Fe, Mn and Cu) nutrients. Soil compaction was determined using a soil pocket penetrometer. Readings were taken at 5 mm from 30 points placed at 1 m intervals and distributed in a randomly established straight line in each plot. Soil loss was determined using a rainfall simulator. Splash cups were filled with soil and placed under the rainfall simulator for 8 minutes per rainstorm at 360 mm-1 intensity. Results showed that at Esigingqini a total of 20 herbaceous species were identified, 17 of which were grasses. About 60percent of the identified grasses were Increaser II species, 15percent were Increaser II and 25percent Decreasers. In terms of palatability, 50percent of the grass species had poor palatability and the remaining had either average or high palatability. At Kwezana, 18 herbaceous species were identified of which 28percent were Decreasers and 61percent were Increaser II species. Half of the species had low palatability and only 28percent had high palatability. In both communal areas, litter cover was highest (P < 0.05) in the non-degraded areas followed by the slightly degraded sites and this was to be expected. Similarly, in both communal areas, there was a shift in species composition of decreasers and perennial grasses being replaced by annual species of poor quality and low palatability as the degradation levels increased. Results on soil chemical properties showed that the non-degraded areas had higher (P < 0.05) pH levels (8.5) than the severely degraded soils (5). A similar trend following a degradation gradient was observed for some soil nutrients such as N, P or Mg. Both the macro and micro nutrient concentrations were relatively higher (P < 0.05) in summer than in winter in both communal areas. Results on physical properties showed that soil loss and compaction were relatively lower (P <0.05) in the slightly degraded and non-degraded sites and increased with severity of degradation. In both communal areas, the severely degraded areas lost soil up to 1.1 t ha-1 due to degradation and the non-degraded areas had a soil loss of less than 0.8 t ha-1. The rate of soil loss between the severely and non-degraded areas is significantly different. The differences may be attributed to the differences in the vegetation cover which influences the rate of erosion on the various degradation gradients. These results suggest that both the grass layer and soil quality is negatively affected by degradation. Therefore, there is a need for land use practices that promote the sustainable use of rangeland resources. Such practices may include rotational grazing, reseeding, veld resting and fire. Measures to control the consequences of degradation and rehabilitate these deteriorated rangelands must be a priority.
- Full Text:
- Date Issued: 2017
- Authors: Maziko, Yonela
- Date: 2017
- Subjects: Communal rangelands -- South Africa -- Eastern Cape Land degradation -- South Africa -- Eastern Cape Vegetation management -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10353/15056 , vital:40159
- Description: This study was conducted to investigate the influence of degradation on vegetation and soil quality in two semi-arid rangelands in the Eastern Cape Province of South Africa. The study was undertaken in Kwezana and Esigingqini communal areas that showed signs id degradation. The assessment incorporated the soil and grass layer in different degradation levels, namely nondegraded, slightly degraded, moderately and severely degraded. The non-degraded has a sufficient amount of vegetation cover and species composition. The slightly degraded site consists of a few visible bare patches. The moderately degraded areas showed signs of active sheet erosion such as pedestals and a higher percentage of bare areas and the severely degraded area is characterized by dongas and gullies, with vast areas that have no ground cover or top fertile soil layer. Using a step point method, herbaceous species composition was determined within three transects from three plots 100 m x 40 m in each degradation level. The aboveground phytomass production was determined by harvesting herbaceous species within four randomly placed 0.25 m2 quadrats in each transect. Bare ground was determined by counting and recording the distance between tufts within each quadrat. Litter cover was determined by visual observation and rated on a scale of 1 to 5. A number of physical and chemical soil properties were used to quantify the effects of degradation on soil quality. To determine soil chemical properties, three soil samples were collected at a depth of 20 cm, in each plot and analyzed for pH, soil macro (N, C, K, P, Ca, Mg) and micro (Zn, Fe, Mn and Cu) nutrients. Soil compaction was determined using a soil pocket penetrometer. Readings were taken at 5 mm from 30 points placed at 1 m intervals and distributed in a randomly established straight line in each plot. Soil loss was determined using a rainfall simulator. Splash cups were filled with soil and placed under the rainfall simulator for 8 minutes per rainstorm at 360 mm-1 intensity. Results showed that at Esigingqini a total of 20 herbaceous species were identified, 17 of which were grasses. About 60percent of the identified grasses were Increaser II species, 15percent were Increaser II and 25percent Decreasers. In terms of palatability, 50percent of the grass species had poor palatability and the remaining had either average or high palatability. At Kwezana, 18 herbaceous species were identified of which 28percent were Decreasers and 61percent were Increaser II species. Half of the species had low palatability and only 28percent had high palatability. In both communal areas, litter cover was highest (P < 0.05) in the non-degraded areas followed by the slightly degraded sites and this was to be expected. Similarly, in both communal areas, there was a shift in species composition of decreasers and perennial grasses being replaced by annual species of poor quality and low palatability as the degradation levels increased. Results on soil chemical properties showed that the non-degraded areas had higher (P < 0.05) pH levels (8.5) than the severely degraded soils (5). A similar trend following a degradation gradient was observed for some soil nutrients such as N, P or Mg. Both the macro and micro nutrient concentrations were relatively higher (P < 0.05) in summer than in winter in both communal areas. Results on physical properties showed that soil loss and compaction were relatively lower (P <0.05) in the slightly degraded and non-degraded sites and increased with severity of degradation. In both communal areas, the severely degraded areas lost soil up to 1.1 t ha-1 due to degradation and the non-degraded areas had a soil loss of less than 0.8 t ha-1. The rate of soil loss between the severely and non-degraded areas is significantly different. The differences may be attributed to the differences in the vegetation cover which influences the rate of erosion on the various degradation gradients. These results suggest that both the grass layer and soil quality is negatively affected by degradation. Therefore, there is a need for land use practices that promote the sustainable use of rangeland resources. Such practices may include rotational grazing, reseeding, veld resting and fire. Measures to control the consequences of degradation and rehabilitate these deteriorated rangelands must be a priority.
- Full Text:
- Date Issued: 2017
Risk analysis assessment of the influence of geological factors on exploration and mining investment alternatives : development of a microcomputer simulation model
- Authors: Mallinson, Clyde A
- Date: 1987
- Subjects: Geology, Economic , Risk assessment -- Computer simulation , Mineral industries -- Economic aspects -- Computer simulation , Prospecting -- Computer simulation
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4904 , http://hdl.handle.net/10962/d1001564
- Description: A microcomputer risk analysis model is developed and used in an exploration - delineation simulation to analyse in particular the effects of geological factors on exploration mining investment alternatives. Analysis of results indicate that geological parameters can have profound effects on such investment alternatives and that the role of the geologist in determining and evaluating the significance of the various geological factors is critical. Simulation examples highlight some of the key geological parameters and show how changes in these parameters influence both the expected mean results and the standard deviations of such means. The risk analysis model provides an ideal means of conveying the importance of the different geoiogical factors on exploration - delineation - mining investment alternatives and may be used as a geological education aid
- Full Text:
- Date Issued: 1987
- Authors: Mallinson, Clyde A
- Date: 1987
- Subjects: Geology, Economic , Risk assessment -- Computer simulation , Mineral industries -- Economic aspects -- Computer simulation , Prospecting -- Computer simulation
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4904 , http://hdl.handle.net/10962/d1001564
- Description: A microcomputer risk analysis model is developed and used in an exploration - delineation simulation to analyse in particular the effects of geological factors on exploration mining investment alternatives. Analysis of results indicate that geological parameters can have profound effects on such investment alternatives and that the role of the geologist in determining and evaluating the significance of the various geological factors is critical. Simulation examples highlight some of the key geological parameters and show how changes in these parameters influence both the expected mean results and the standard deviations of such means. The risk analysis model provides an ideal means of conveying the importance of the different geoiogical factors on exploration - delineation - mining investment alternatives and may be used as a geological education aid
- Full Text:
- Date Issued: 1987
A review of landscape development and erosion cycles in southern Africa
- Authors: Corbett, Dudley Harold
- Date: 1979
- Subjects: Landscape changes -- Africa, Southern Erosion -- Africa, Southern
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4899 , http://hdl.handle.net/10962/d1001558
- Description: The study of landforms has, to a large extent been neglected by exploration geologists. This is surprising because field geology is intimately related with landforms of one type or another. An understanding of the geomorphological history of a particular area will assist in the appreciation of the surficial environment, the processes of weathering, soil formation and duricrust (calcrete , ferricrete silcrete) formation. This has direct application in planning and interpreting geochemical sampling programmes. In order to understand the evolution of landforms it is necessary to study slope development in some detail. Agents of erosion and denudation constantly at work, remove detritus from hillcrests down to the drainage lines which form the local base level from whence material is transported down to the sea. The system is one of dynamic equilibrium, and the concept of grade is important in understanding hillslope evolution. Thus Section (1) of this discussion deals with past and present theories of hillslope evolution. Very little work has been done recently on the geomorphology of Southern Africa and the presently accepted classification of land surfaces is based on the work of Prof. L.C. King (numerous publications). However, through the more recent work of De Swardt and Bennet (1974) on the geomorphology of Natal, it is apparent that the present system of landform classification in Southern Africa requires widespread revision. The relationship of erosion cycles in Southern Africa to the late Jurassic-early Cretaceous break-up of Gondwanaland has received insufficient attention in the past. Erosion cycles bear an intimate relationship to offshore Cretaceous and Tertiary-Recent sedimentation. Valuable information on these sediments has only recently become available as a result of offshore exploration for oil. The findings of De Swardt and Bennet (1974) are summarized in Section (2) and the present land surface classification of Southern Africa is reviewed. Finally, some suggestions on a new interpretation of land surfaces in Southern Africa are given.
- Full Text:
- Date Issued: 1979
- Authors: Corbett, Dudley Harold
- Date: 1979
- Subjects: Landscape changes -- Africa, Southern Erosion -- Africa, Southern
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4899 , http://hdl.handle.net/10962/d1001558
- Description: The study of landforms has, to a large extent been neglected by exploration geologists. This is surprising because field geology is intimately related with landforms of one type or another. An understanding of the geomorphological history of a particular area will assist in the appreciation of the surficial environment, the processes of weathering, soil formation and duricrust (calcrete , ferricrete silcrete) formation. This has direct application in planning and interpreting geochemical sampling programmes. In order to understand the evolution of landforms it is necessary to study slope development in some detail. Agents of erosion and denudation constantly at work, remove detritus from hillcrests down to the drainage lines which form the local base level from whence material is transported down to the sea. The system is one of dynamic equilibrium, and the concept of grade is important in understanding hillslope evolution. Thus Section (1) of this discussion deals with past and present theories of hillslope evolution. Very little work has been done recently on the geomorphology of Southern Africa and the presently accepted classification of land surfaces is based on the work of Prof. L.C. King (numerous publications). However, through the more recent work of De Swardt and Bennet (1974) on the geomorphology of Natal, it is apparent that the present system of landform classification in Southern Africa requires widespread revision. The relationship of erosion cycles in Southern Africa to the late Jurassic-early Cretaceous break-up of Gondwanaland has received insufficient attention in the past. Erosion cycles bear an intimate relationship to offshore Cretaceous and Tertiary-Recent sedimentation. Valuable information on these sediments has only recently become available as a result of offshore exploration for oil. The findings of De Swardt and Bennet (1974) are summarized in Section (2) and the present land surface classification of Southern Africa is reviewed. Finally, some suggestions on a new interpretation of land surfaces in Southern Africa are given.
- Full Text:
- Date Issued: 1979
Prioritising native fish populations for conservation using genetics in the Groot Marico catchment, North West Province, South Africa
- Authors: Van der Walt, Kerry-Ann
- Date: 2014
- Subjects: Native fishes Fishery management -- South Africa -- North West Fish populations Fishes -- Conservation -- South Africa -- Western Cape
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/69102 , vital:29390
- Description: The Groot Marico catchment in the North West Province is a National Freshwater Ecosystem Priority Area (NFEPA) because it represents unique landscape features with unique biodiversity that are considered to be of special ecological significance. Three native freshwater species Amphilius uranoscopus, Chiloglanis pretoriae and Barbus motebensis, have high local conservation importance and B. motebensis is endemic to the catchment and is IUCN-listed as vulnerable. The main objective of this study is to contribute towards the effective conservation of these three species in the Groot Marico River system by assessing their genetic structure to determine whether tributary populations of the three species comprise of one genetic population or whether they are divided into genetically distinct subpopulations, in order to prioritise areas for conservation. The central null hypothesis was that there is no genetic differentiation between tributary populations (i.e., panmixia) of B. motebensis, A. uranoscopus and C. pretoriae in the Groot Marico catchment, North West Province. In total, 80 individuals per species were collected, targeting at least 10 individuals per population from a total of eight populations (seven tributaries and the Groot Marico main stem) and across the study area. Samples were collected by electrofishing and specimens were euthanized using an overdose of clove oil. A sample of muscle tissue was removed for genetic evaluation and the remainder of the specimens served as voucher specimens. For the genetic evaluation, mitochondrial (ND2, cyt b) and nuclear (S7) genes were used. Genetic techniques used were DNA extraction, polymerase chain reaction (PCR), purification and sequencing. From the 240 individuals collected, 123 sequences for B. motebensis, 111 sequences for A. uranoscopus and 103 sequences for C. pretoriae were analysed across all three genes. Statistical analysis included looking at cleaned sequences in order to obtain models using MODELTEST (version 3.06). Population structuring and phylogeographic analysis was performed in Arlequin (version 2000), TCS (version 1.2.1) and PAUP*. Results indicated that for B. motebensis the null hypothesis could be rejected as there were two distinct lineages (the Draai and Eastern lineages) that demonstrated significant divergence in both the ND2 and S7 genes, suggesting historical isolation. The low divergence in the mitochondrial cytochrome b gene (0% < D < 0.8%) suggests that this isolation is not very old and is probably not comparable to species level differentiation. The null hypothesis was also rejected for A. uranoscopus as there were also significant levels of differentiation between tributary populations resulting in the identification of two lineages (the Ribbok and Western lineages). However, for C. pretoriae, the null hypothesis could not be rejected as there was no genetic differentiation between tributary populations i.e., one panmictic population. Therefore, due to each species showing different genetic structuring within the tributary populations, more than one priority area for conservation needs to be implemented. These priority areas of conservation where therefore evaluated based on the current conservation status of the species (B. motebensis being vulnerable on the IUCN Red List), the number of Evolutionary Significant Units for each species and the overall genetic diversity of all three species in the Groot Marico catchment. In total, four tributary populations were conservation priorities areas, these were the Draai, Vanstraatens, Ribbok and Kaaloog tributaries. The Draai, Vanstraatens and Kaaloog tributaries were selected as priority areas for B. motebensis (B. motebensis is considered to be the most vulnerable of all three species). The Draai tributary was selected due to the B. motebensis population within the tributary showing isolation from the rest of the tributary populations. In order to conserve B. motebensis from the Southern lineage, the Vanstraatens and Kaaloog tributaries were selected. Reasons for selecting these two specific tributaries within the Southern lineage were that the Vanstraatens tributary had unique alleles (three Evolutionary Significant Units) for B. motebensis and the Kaaloog tributary had high genetic diversity (HD = 0.889, ND2 gene) when compared to the other tributary populations. The Ribbok and Vanstraatens tributaries were selected as priority areas for the conservation of A. uranoscopus. The Ribbok tributary was selected as it showed isolation from the rest of the tributary populations, as seen with the Draai tributary (B. motebensis) and the Vanstraatens tributary was selected to represent the Western lineage as it had the highest diversity for both genes (ND2 and S7). The Ribbok tributary has the highest prioritisation when compared to the Vanstraatens tributary. Chiloglanis pretoriae occurs within the Draai, Vanstraatens, Ribbok and Kaaloog tributaries, therefore by prioritising these tributaries for conservation, C. pretoriae will in turn be conserved.
- Full Text:
- Date Issued: 2014
- Authors: Van der Walt, Kerry-Ann
- Date: 2014
- Subjects: Native fishes Fishery management -- South Africa -- North West Fish populations Fishes -- Conservation -- South Africa -- Western Cape
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/69102 , vital:29390
- Description: The Groot Marico catchment in the North West Province is a National Freshwater Ecosystem Priority Area (NFEPA) because it represents unique landscape features with unique biodiversity that are considered to be of special ecological significance. Three native freshwater species Amphilius uranoscopus, Chiloglanis pretoriae and Barbus motebensis, have high local conservation importance and B. motebensis is endemic to the catchment and is IUCN-listed as vulnerable. The main objective of this study is to contribute towards the effective conservation of these three species in the Groot Marico River system by assessing their genetic structure to determine whether tributary populations of the three species comprise of one genetic population or whether they are divided into genetically distinct subpopulations, in order to prioritise areas for conservation. The central null hypothesis was that there is no genetic differentiation between tributary populations (i.e., panmixia) of B. motebensis, A. uranoscopus and C. pretoriae in the Groot Marico catchment, North West Province. In total, 80 individuals per species were collected, targeting at least 10 individuals per population from a total of eight populations (seven tributaries and the Groot Marico main stem) and across the study area. Samples were collected by electrofishing and specimens were euthanized using an overdose of clove oil. A sample of muscle tissue was removed for genetic evaluation and the remainder of the specimens served as voucher specimens. For the genetic evaluation, mitochondrial (ND2, cyt b) and nuclear (S7) genes were used. Genetic techniques used were DNA extraction, polymerase chain reaction (PCR), purification and sequencing. From the 240 individuals collected, 123 sequences for B. motebensis, 111 sequences for A. uranoscopus and 103 sequences for C. pretoriae were analysed across all three genes. Statistical analysis included looking at cleaned sequences in order to obtain models using MODELTEST (version 3.06). Population structuring and phylogeographic analysis was performed in Arlequin (version 2000), TCS (version 1.2.1) and PAUP*. Results indicated that for B. motebensis the null hypothesis could be rejected as there were two distinct lineages (the Draai and Eastern lineages) that demonstrated significant divergence in both the ND2 and S7 genes, suggesting historical isolation. The low divergence in the mitochondrial cytochrome b gene (0% < D < 0.8%) suggests that this isolation is not very old and is probably not comparable to species level differentiation. The null hypothesis was also rejected for A. uranoscopus as there were also significant levels of differentiation between tributary populations resulting in the identification of two lineages (the Ribbok and Western lineages). However, for C. pretoriae, the null hypothesis could not be rejected as there was no genetic differentiation between tributary populations i.e., one panmictic population. Therefore, due to each species showing different genetic structuring within the tributary populations, more than one priority area for conservation needs to be implemented. These priority areas of conservation where therefore evaluated based on the current conservation status of the species (B. motebensis being vulnerable on the IUCN Red List), the number of Evolutionary Significant Units for each species and the overall genetic diversity of all three species in the Groot Marico catchment. In total, four tributary populations were conservation priorities areas, these were the Draai, Vanstraatens, Ribbok and Kaaloog tributaries. The Draai, Vanstraatens and Kaaloog tributaries were selected as priority areas for B. motebensis (B. motebensis is considered to be the most vulnerable of all three species). The Draai tributary was selected due to the B. motebensis population within the tributary showing isolation from the rest of the tributary populations. In order to conserve B. motebensis from the Southern lineage, the Vanstraatens and Kaaloog tributaries were selected. Reasons for selecting these two specific tributaries within the Southern lineage were that the Vanstraatens tributary had unique alleles (three Evolutionary Significant Units) for B. motebensis and the Kaaloog tributary had high genetic diversity (HD = 0.889, ND2 gene) when compared to the other tributary populations. The Ribbok and Vanstraatens tributaries were selected as priority areas for the conservation of A. uranoscopus. The Ribbok tributary was selected as it showed isolation from the rest of the tributary populations, as seen with the Draai tributary (B. motebensis) and the Vanstraatens tributary was selected to represent the Western lineage as it had the highest diversity for both genes (ND2 and S7). The Ribbok tributary has the highest prioritisation when compared to the Vanstraatens tributary. Chiloglanis pretoriae occurs within the Draai, Vanstraatens, Ribbok and Kaaloog tributaries, therefore by prioritising these tributaries for conservation, C. pretoriae will in turn be conserved.
- Full Text:
- Date Issued: 2014
Immobilisation of electric eel acetylcholinesterase on nanofibres electrospun from a nylon and chitosan blend
- Authors: Mafuma, Tendai Simbarashe
- Date: 2013
- Subjects: Acetylcholinesterase Acetylcholinesterase -- Inhibitors Electric eel Biosensors Immobilized enzymes Pesticides -- Environmental aspects Pesticides -- Toxicology Nylon Chitosan Nanofibers Electrospinning
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3886 , http://hdl.handle.net/10962/d1001620
- Description: Organophosphates and carbamates are potent inhibitors of the neurotransmitter acetylcholinesterase. This inhibition results in the blocking of nerve signal transference into the post synaptic neuron leading to loss of muscle action and death. Because of the universal mechanisms of signal transduction in animals, these inhibitors have been widely used as agricultural pesticides as well as chemical warfare agents (nerve agents). Health issues associated with pesticide usage result from the fact that both the pesticides and their breakdown products often end up in water and food sources as well as in the soil. As a result, there has been an increase in the number of studies aimed at the detection of these pesticides in the environment. One popular research area is enzyme based biosensor construction. Some important criteria for consideration during the construction of biosensors are the importance of a suitable solid support as well as the enzyme immobilisation method. Recently, there has been increased interest in using nano-scale material e.g. using nanoparticles as enzyme support material. This is largely due to their advantages such as large surface area to volume ratio as well as reduced mass transfer resistance. Electrospinning is a straight forward and cost effective method for producing nanofibres from any soluble polymer(s). The applications of electrospun nanofibres have been reported in clinical studies, biofuel production as well as bioremediation. In this study two polymers were selected: nylon for its mechanical stability and chitosan for its biocompatibility and hydrophilicity, for the fabrication of electrospun nanofibres which would function as immobilisation support material for acetylcholinesterase. The first objective of this study was to electrospin nanofibres from a nylon-6 and chitosan blend solution. A binary solvent system consisting of formic acid and acetic acid (50:50) successfully dissolved and blended the polymers which were subsequently electrospun. Scanning electron microscopy characterisation of the nanofibres showed that (i) a nylon-6: chitosan ratio of 16%: 3% resulted in the formation of bead free nanofibres and (ii) the fibres were collected in non-woven mats characterised by different size nanofibres with average diameters of 250 nm for the main fibres and 40 nm for the smaller nanofibres. Fourier transform infra-red (FT-IR) analysis of the nanofibres indicated that a new product had been formed during the blending of the two polymers. The second aim of the study was to carry out a facile immobilisation of electric eel acetylcholinesterase via glutaraldehyde (GA) cross-linking. Glutaraldehyde solution 5% (v/v) resulted in the immobilisation of 0.334 mg/cm² of acetylcholinesterase onto the nanofibres. The immobilisation procedure was optimised with reference to acetylcholinestease and crosslinker concentrations, incubation time and the cross-linking method. A comparative investigation into the optimum pH and temperature conditions, pH and thermal stabilities, substrate and inhibition kinetics was then carried out on free and immobilised acetylcholinesterase. The final objective of this study was to determine the storage stabilities of the immobilised and free enzymes as well as the reusability characteristics of the immobilised acetylcholinesterase. Several conclusions were drawn from this study. Acetylcholinesterase was successfully immobilised onto the surface of nylon-6:chitosan nanofibres with retention of its activity. There was a shift in the pH optimum of the immobilised acetylcholineseterase by 0.5 units towards a neutral pH. Although both free and immobilised acetylcholinesterase exhibited the same optimum temperature, immobilised acetylcholinesterase showed enhanced thermal stability. In terms of pH stability, immobilised acetylcholinesterase showed greater stability at acidic pH whilst free acetylcholinesterase was more stable under alkaline pH conditions. Relative to free acetylcholinesterase, the immobilised enzyme showed considerable storage stability retaining ~50% of its activity when stored for 49 days at 4°C. Immobilised acetylcholinesterase also retained > 20% of its initial activity after 9 consecutive reuse cycles. When exposed to fixed concentrations of carbofuran or demeton-S-methyl sulfone, immobilised acetylcholinesterase showed similar inhibition characteristics to that of the free enzyme. The decrease in enzyme activity observed after immobilisation to the nanofibres may have been due to several reasons which include some enzyme molecules being immobilised in structural conformations which reduced substrate access to the catalytic site, participation of the catalytic residues in immobilisation and enzyme denaturation due to the reaction conditions used for acetylcholinesterase immobilisation. Similar observations have been widely reported in literature and this is one of the major drawbacks of enzyme immobilisation. In conclusion, nylon-6:chitosan electrospun nanofibres were shown to be suitable supports for facile acetylcholinesterase immobilisation and the immobilised enzyme has potential for use in pesticide detection. Future recommendations for this study include a comparative study of the GA cross-linking method for AChE immobilisation which will lead to more intensely bound enzyme molecules to prevent non-specific binding. An investigation into the effect of inhibitors on stored immobilised AChE, as well as reactivation and reuse studies, may also be useful for determining the cost-effectiveness of reusing immobilised AChE for pesticide detection in environmental water samples. Several models have been designed for the determination of the kinetic parameters for immobilised enzymes. These take into account the mass transfer resistance as well as the overall charge of the immobilisation matrix. The use of these models to analyse experimental data will give a clear understanding of the effects of immobilisation on enzyme activity
- Full Text:
- Date Issued: 2013
- Authors: Mafuma, Tendai Simbarashe
- Date: 2013
- Subjects: Acetylcholinesterase Acetylcholinesterase -- Inhibitors Electric eel Biosensors Immobilized enzymes Pesticides -- Environmental aspects Pesticides -- Toxicology Nylon Chitosan Nanofibers Electrospinning
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3886 , http://hdl.handle.net/10962/d1001620
- Description: Organophosphates and carbamates are potent inhibitors of the neurotransmitter acetylcholinesterase. This inhibition results in the blocking of nerve signal transference into the post synaptic neuron leading to loss of muscle action and death. Because of the universal mechanisms of signal transduction in animals, these inhibitors have been widely used as agricultural pesticides as well as chemical warfare agents (nerve agents). Health issues associated with pesticide usage result from the fact that both the pesticides and their breakdown products often end up in water and food sources as well as in the soil. As a result, there has been an increase in the number of studies aimed at the detection of these pesticides in the environment. One popular research area is enzyme based biosensor construction. Some important criteria for consideration during the construction of biosensors are the importance of a suitable solid support as well as the enzyme immobilisation method. Recently, there has been increased interest in using nano-scale material e.g. using nanoparticles as enzyme support material. This is largely due to their advantages such as large surface area to volume ratio as well as reduced mass transfer resistance. Electrospinning is a straight forward and cost effective method for producing nanofibres from any soluble polymer(s). The applications of electrospun nanofibres have been reported in clinical studies, biofuel production as well as bioremediation. In this study two polymers were selected: nylon for its mechanical stability and chitosan for its biocompatibility and hydrophilicity, for the fabrication of electrospun nanofibres which would function as immobilisation support material for acetylcholinesterase. The first objective of this study was to electrospin nanofibres from a nylon-6 and chitosan blend solution. A binary solvent system consisting of formic acid and acetic acid (50:50) successfully dissolved and blended the polymers which were subsequently electrospun. Scanning electron microscopy characterisation of the nanofibres showed that (i) a nylon-6: chitosan ratio of 16%: 3% resulted in the formation of bead free nanofibres and (ii) the fibres were collected in non-woven mats characterised by different size nanofibres with average diameters of 250 nm for the main fibres and 40 nm for the smaller nanofibres. Fourier transform infra-red (FT-IR) analysis of the nanofibres indicated that a new product had been formed during the blending of the two polymers. The second aim of the study was to carry out a facile immobilisation of electric eel acetylcholinesterase via glutaraldehyde (GA) cross-linking. Glutaraldehyde solution 5% (v/v) resulted in the immobilisation of 0.334 mg/cm² of acetylcholinesterase onto the nanofibres. The immobilisation procedure was optimised with reference to acetylcholinestease and crosslinker concentrations, incubation time and the cross-linking method. A comparative investigation into the optimum pH and temperature conditions, pH and thermal stabilities, substrate and inhibition kinetics was then carried out on free and immobilised acetylcholinesterase. The final objective of this study was to determine the storage stabilities of the immobilised and free enzymes as well as the reusability characteristics of the immobilised acetylcholinesterase. Several conclusions were drawn from this study. Acetylcholinesterase was successfully immobilised onto the surface of nylon-6:chitosan nanofibres with retention of its activity. There was a shift in the pH optimum of the immobilised acetylcholineseterase by 0.5 units towards a neutral pH. Although both free and immobilised acetylcholinesterase exhibited the same optimum temperature, immobilised acetylcholinesterase showed enhanced thermal stability. In terms of pH stability, immobilised acetylcholinesterase showed greater stability at acidic pH whilst free acetylcholinesterase was more stable under alkaline pH conditions. Relative to free acetylcholinesterase, the immobilised enzyme showed considerable storage stability retaining ~50% of its activity when stored for 49 days at 4°C. Immobilised acetylcholinesterase also retained > 20% of its initial activity after 9 consecutive reuse cycles. When exposed to fixed concentrations of carbofuran or demeton-S-methyl sulfone, immobilised acetylcholinesterase showed similar inhibition characteristics to that of the free enzyme. The decrease in enzyme activity observed after immobilisation to the nanofibres may have been due to several reasons which include some enzyme molecules being immobilised in structural conformations which reduced substrate access to the catalytic site, participation of the catalytic residues in immobilisation and enzyme denaturation due to the reaction conditions used for acetylcholinesterase immobilisation. Similar observations have been widely reported in literature and this is one of the major drawbacks of enzyme immobilisation. In conclusion, nylon-6:chitosan electrospun nanofibres were shown to be suitable supports for facile acetylcholinesterase immobilisation and the immobilised enzyme has potential for use in pesticide detection. Future recommendations for this study include a comparative study of the GA cross-linking method for AChE immobilisation which will lead to more intensely bound enzyme molecules to prevent non-specific binding. An investigation into the effect of inhibitors on stored immobilised AChE, as well as reactivation and reuse studies, may also be useful for determining the cost-effectiveness of reusing immobilised AChE for pesticide detection in environmental water samples. Several models have been designed for the determination of the kinetic parameters for immobilised enzymes. These take into account the mass transfer resistance as well as the overall charge of the immobilisation matrix. The use of these models to analyse experimental data will give a clear understanding of the effects of immobilisation on enzyme activity
- Full Text:
- Date Issued: 2013
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