The structure and function of the gametes of the striped field mouse (Rhabdomys pumilio)
- Authors: Tinney, Gregory Michael
- Date: 2000
- Subjects: Gametes , Microtus , Mice
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5774 , http://hdl.handle.net/10962/d1005462 , Gametes , Microtus , Mice
- Description: This study was initiated in an attempt toadd the four-striped field mouse (Rhabdomys pumilio) to the small group of mammals that are used to study and develop assisted reproductive technologies. To accomplish this, a complete knowledge of both the male and female gametes was necessary, as well as a repeatable and successful IVF protocol. The gametes of the field mouse, both the falciformspermand the mature oocytes, were shown to be similar in several respects to those of many other rodent species. The induction of both the capacitated state and the acrosome reaction in the sperm were readily achieved. The success of retrieving mature oocytes was very low, with the rare occurrence of polar bodies within ovulated oocytes. The difficulties with oocytes was either related to the lack of a regular oestrous cycle, or to the hormonal regime used to induce superovulation. Although several protocols were used to accomplish superovulation in this species, it was seldom achieved. In in vitro fertilization trials, sperm were found to attach to both the cumulus oophorus and the zona pellucida of the oocytes. However, fertilization was never accomplished. This failure to achieve fertilization was probably related to the immaturity of the oocytes. Without further studies on Rhabdomys pumilio and further attempts at accomplishing in vitro fertilization, it would seem that this species would not easily be introduced as an animalmodelfor assisted reproduction.
- Full Text:
- Authors: Tinney, Gregory Michael
- Date: 2000
- Subjects: Gametes , Microtus , Mice
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5774 , http://hdl.handle.net/10962/d1005462 , Gametes , Microtus , Mice
- Description: This study was initiated in an attempt toadd the four-striped field mouse (Rhabdomys pumilio) to the small group of mammals that are used to study and develop assisted reproductive technologies. To accomplish this, a complete knowledge of both the male and female gametes was necessary, as well as a repeatable and successful IVF protocol. The gametes of the field mouse, both the falciformspermand the mature oocytes, were shown to be similar in several respects to those of many other rodent species. The induction of both the capacitated state and the acrosome reaction in the sperm were readily achieved. The success of retrieving mature oocytes was very low, with the rare occurrence of polar bodies within ovulated oocytes. The difficulties with oocytes was either related to the lack of a regular oestrous cycle, or to the hormonal regime used to induce superovulation. Although several protocols were used to accomplish superovulation in this species, it was seldom achieved. In in vitro fertilization trials, sperm were found to attach to both the cumulus oophorus and the zona pellucida of the oocytes. However, fertilization was never accomplished. This failure to achieve fertilization was probably related to the immaturity of the oocytes. Without further studies on Rhabdomys pumilio and further attempts at accomplishing in vitro fertilization, it would seem that this species would not easily be introduced as an animalmodelfor assisted reproduction.
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The structure and microbiology of floating sulphide oxidising biofilms
- Authors: Gilfillan, Joanne Criseyde
- Date: 2000
- Subjects: Biofilms , Sulfides
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3903 , http://hdl.handle.net/10962/d1003962 , Biofilms , Sulfides
- Description: Floating sulphur biofilms are observed as surface layers in numerous aquatic sulphide-rich environments, and apparently play an important role in the cycling of sulphur in its various oxidation states. In addition to the conversion of sulphide to sulphur and/or sulphate species, it has been suspected that subsequent reduction back to sulphide may occur within the floating sulphur biofi1m in organic-rich environments. The use of sulphur biofilms for the harvesting of elemental sulphur from wastewater treatment systems has also been suggested. There is, however, little documented information in the literature on the structure of floating sulphur biofilms, or the microbial species responsible for their occurrence. In this study, floating sulphur biofilms were generated in a continuous flow baflle reactor and their structure was examined using scanning electron microscopy. It was found that they occur as layered structures with morphologically distinct bacterial forms present in different layers of the biofilm. The biofilpl structure was also found to be dynamic, with structural changes observed as feed conditions were altered. An enriched culture derived from the biofi1m demonstrated rates of sulphide oxidation comparable to values reported in the literature for liquid culture systems. The microbiology of the biofi1m was studied using traditional plate culture techniques and analysis ofrRNA genes. Identification of plate culture isolates as representatives of the biofi1m community proved to be limited, leading to a PeR-based cloning approach. The majority of the organisms present in the sulphur biofi1m were classified as species in the genus ~eudomonas, and a number of other bacterial species whose sulphide oxidising capacity has been noted previously. Surprisingly, only 2% of the clone library consisted of Thiobacillus spp., and no sulphate reducing bacteria were identified in the biofilm at all. These results indicate that in organic sulphide-rich environments facultative chemolithoheterotrophic bacterial forms predominate in floating sulphur biofilms, and that the complete biological cycling of sulphur may not occur in these systems.
- Full Text:
- Authors: Gilfillan, Joanne Criseyde
- Date: 2000
- Subjects: Biofilms , Sulfides
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3903 , http://hdl.handle.net/10962/d1003962 , Biofilms , Sulfides
- Description: Floating sulphur biofilms are observed as surface layers in numerous aquatic sulphide-rich environments, and apparently play an important role in the cycling of sulphur in its various oxidation states. In addition to the conversion of sulphide to sulphur and/or sulphate species, it has been suspected that subsequent reduction back to sulphide may occur within the floating sulphur biofi1m in organic-rich environments. The use of sulphur biofilms for the harvesting of elemental sulphur from wastewater treatment systems has also been suggested. There is, however, little documented information in the literature on the structure of floating sulphur biofilms, or the microbial species responsible for their occurrence. In this study, floating sulphur biofilms were generated in a continuous flow baflle reactor and their structure was examined using scanning electron microscopy. It was found that they occur as layered structures with morphologically distinct bacterial forms present in different layers of the biofilm. The biofilpl structure was also found to be dynamic, with structural changes observed as feed conditions were altered. An enriched culture derived from the biofi1m demonstrated rates of sulphide oxidation comparable to values reported in the literature for liquid culture systems. The microbiology of the biofi1m was studied using traditional plate culture techniques and analysis ofrRNA genes. Identification of plate culture isolates as representatives of the biofi1m community proved to be limited, leading to a PeR-based cloning approach. The majority of the organisms present in the sulphur biofi1m were classified as species in the genus ~eudomonas, and a number of other bacterial species whose sulphide oxidising capacity has been noted previously. Surprisingly, only 2% of the clone library consisted of Thiobacillus spp., and no sulphate reducing bacteria were identified in the biofilm at all. These results indicate that in organic sulphide-rich environments facultative chemolithoheterotrophic bacterial forms predominate in floating sulphur biofilms, and that the complete biological cycling of sulphur may not occur in these systems.
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The use of indigenous macroinvertebrates and Daphnia pulex in acute toxicity testing
- Authors: Everitt, Victoria Jane
- Date: 2000
- Subjects: Water -- Pollution -- Toxicology , Toxicity testing , Daphnia pulex
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5795 , http://hdl.handle.net/10962/d1005483
- Description: Aquatic toxicology has been identified as a valuable tool in the identification and management of chemical pollution in aquatic ecosystems. Standardised methodologies for acute aquatic bioassays have been adopted from international agencies. As a result of these standard methods, the use of laboratory cultured organisms for toxicity testing has been more popular than that of indigenous field-caught organisms. Included in these adopted methods are those for the cultured crustacean Daphnia pUlex. D.pulex is adapted to living in standing water and the suitability of this species to determine toxic effects for South African riverine environments, which are largely flowing, has been questioned. Thus this thesis is a case-study ofthe use of D.pulex and indigenous site-specific macroinvertebrates as toxicity test organisms for setting acute water quality guidelines to protect aquatic ecosystems. The study highlights site-specific problems such as reference sites and organism identification. The acute tolerance of selected indigenous invertebrates was compared to that of D. pulex, using both a single-substance reference toxicant (zinc) and selected whole efiluents. The significance of source population and culture age as a potential source of biological variability between D.pulex cultures was also investigated. D.pulex cultures have been initiated in South Africa from females collected from a number of different local populations; also it is assumed that no genetic change (due to mutation) occurs within a D.pulex culture over time. In order to establish if source population and culture age are a source of biological variability between D.pulex experiments, the acute tolerJuce to zinc of two different D.pulex populations and three different generations within a population were compared. Due to experimental variability results were inconclusive, and differences in tolerance as a result of population difference or culture age could not be determined with confidence. The acute tolerance of D.pulex to a single reference chemical (zinc) and selected whole efiluents was compared to that of selected indigenous invertebrates. Acute 48 h D.pulex zinc tolerance (LC50 range: 0.22 - 0.60 mg/l Zn) was found to be more sensitive than acute 96 h tolerances shown by mayfly species A.fconurus peringueyi (Heptageniidae) (LC50: 17.42 mg/l Zn), Euthrauluselegans (Leptophlebiidae) (LC50: 0.98 mg/IZn), Ba~tidae (LC50: 0.94mg/IZn) and shrimp, Caradina nilotica (Atyidae) (LC50: 3.17 mg/l Zn). This result suggests that guidelines for zinc set using D.pulex will protect the selected indigenous invertebrates. Selected whole eftluents were not acutely toxic to either D.pulex or selected indigenous invertebrates. These experiments were used as a case study for method development regarding the comparative use of D.pulex and indigenous invertebrates in acute whole eftluent toxicity testing. Finally, it is recommended that a suite of indigenous organisms (e.g. macroinvertebrates, fish and algae), as well as laboratory cultured D.pulex, be used in the initial setting of guidelines and that D.pulex be used for routine compliance monitoring. It is futher recommended that a suite of available monitoring methods, such as chemical and biomonitoring methodologies, be used in conjuction with toxicity testing in water quality management.
- Full Text:
- Authors: Everitt, Victoria Jane
- Date: 2000
- Subjects: Water -- Pollution -- Toxicology , Toxicity testing , Daphnia pulex
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5795 , http://hdl.handle.net/10962/d1005483
- Description: Aquatic toxicology has been identified as a valuable tool in the identification and management of chemical pollution in aquatic ecosystems. Standardised methodologies for acute aquatic bioassays have been adopted from international agencies. As a result of these standard methods, the use of laboratory cultured organisms for toxicity testing has been more popular than that of indigenous field-caught organisms. Included in these adopted methods are those for the cultured crustacean Daphnia pUlex. D.pulex is adapted to living in standing water and the suitability of this species to determine toxic effects for South African riverine environments, which are largely flowing, has been questioned. Thus this thesis is a case-study ofthe use of D.pulex and indigenous site-specific macroinvertebrates as toxicity test organisms for setting acute water quality guidelines to protect aquatic ecosystems. The study highlights site-specific problems such as reference sites and organism identification. The acute tolerance of selected indigenous invertebrates was compared to that of D. pulex, using both a single-substance reference toxicant (zinc) and selected whole efiluents. The significance of source population and culture age as a potential source of biological variability between D.pulex cultures was also investigated. D.pulex cultures have been initiated in South Africa from females collected from a number of different local populations; also it is assumed that no genetic change (due to mutation) occurs within a D.pulex culture over time. In order to establish if source population and culture age are a source of biological variability between D.pulex experiments, the acute tolerJuce to zinc of two different D.pulex populations and three different generations within a population were compared. Due to experimental variability results were inconclusive, and differences in tolerance as a result of population difference or culture age could not be determined with confidence. The acute tolerance of D.pulex to a single reference chemical (zinc) and selected whole efiluents was compared to that of selected indigenous invertebrates. Acute 48 h D.pulex zinc tolerance (LC50 range: 0.22 - 0.60 mg/l Zn) was found to be more sensitive than acute 96 h tolerances shown by mayfly species A.fconurus peringueyi (Heptageniidae) (LC50: 17.42 mg/l Zn), Euthrauluselegans (Leptophlebiidae) (LC50: 0.98 mg/IZn), Ba~tidae (LC50: 0.94mg/IZn) and shrimp, Caradina nilotica (Atyidae) (LC50: 3.17 mg/l Zn). This result suggests that guidelines for zinc set using D.pulex will protect the selected indigenous invertebrates. Selected whole eftluents were not acutely toxic to either D.pulex or selected indigenous invertebrates. These experiments were used as a case study for method development regarding the comparative use of D.pulex and indigenous invertebrates in acute whole eftluent toxicity testing. Finally, it is recommended that a suite of indigenous organisms (e.g. macroinvertebrates, fish and algae), as well as laboratory cultured D.pulex, be used in the initial setting of guidelines and that D.pulex be used for routine compliance monitoring. It is futher recommended that a suite of available monitoring methods, such as chemical and biomonitoring methodologies, be used in conjuction with toxicity testing in water quality management.
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The utilisation of Asolla filiculoides Lam. as a biofertiliser under dryland conditions
- Authors: Kiguli, Lillian Nakibuuka
- Date: 2000
- Subjects: Asolla as fertilizer
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4206 , http://hdl.handle.net/10962/d1003775 , Asolla as fertilizer
- Description: The response of wheat to soil fertilised with varying quantities of the water fern Azolla filiculoides was investigated. Experiments were conducted to differentiate between the effects of increased soil mineral status and water status. In the preliminary investigation, experiments were carried out in the greenhouse using potted wheat grown in sand with varying proportions of A. filiculoides that had been subjected tovarious pre-treatments. The pre-treatments were fresh, dry and heated A. filiculoides applied at 20%, 50% and 80% volume per 3000 ml. There were significant differences in the measured growth parameters between the plants grown in the various treatments. In addition, the grain yield of wheat plants varied with the different treatments. Results of the preliminary study showed that the addition of heated and dried A. filiculoides resulted in significantly better growth than the addition of fresh A. filiculoides in sand. For fresh biomass, grain weights, Leaf area ratio (LAR) and relative growth rate (RGR), the performance of dried A. filiculoides was as good as that of the heated A. filiculoides. Productivity of wheat in the heated treatments increased significantly with increasing proportion of A. filiculoides added to sand, while in dry treatments there were no significant increases in productivity in the preliminary study. This supported the hypothesis that A. filiculoides, a notorious water weed can be put to agricultural use under dryland conditions in poor nutrient soils. Further investigations using dried A. filiculoides in sand and topsoil showed that the use of the same amounts of the dried fern made no significant short term impact on topsoil grown winter wheat but significantly improved the productivity of wheat in sand. Results showed that the addition of dried 20% Azolla to sand improved the soil fertility to levels equalling the quality of the control topsoil, but the addition of 80% Azolla to sand led to significantly greater wheat productivity than all other treatments. The addition of dried 20% Azolla (8.14 × 10^3 kg ha^(-1)) in sand produced as much wheat biomass as the addition of the recommended NPK fertiliser (30 kg N ha^(-1)) to sand. A comparison between the topsoil and sand-grown plants showed differences in flowering time but these had no effect on the final grain and above ground biomass.
- Full Text:
- Authors: Kiguli, Lillian Nakibuuka
- Date: 2000
- Subjects: Asolla as fertilizer
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4206 , http://hdl.handle.net/10962/d1003775 , Asolla as fertilizer
- Description: The response of wheat to soil fertilised with varying quantities of the water fern Azolla filiculoides was investigated. Experiments were conducted to differentiate between the effects of increased soil mineral status and water status. In the preliminary investigation, experiments were carried out in the greenhouse using potted wheat grown in sand with varying proportions of A. filiculoides that had been subjected tovarious pre-treatments. The pre-treatments were fresh, dry and heated A. filiculoides applied at 20%, 50% and 80% volume per 3000 ml. There were significant differences in the measured growth parameters between the plants grown in the various treatments. In addition, the grain yield of wheat plants varied with the different treatments. Results of the preliminary study showed that the addition of heated and dried A. filiculoides resulted in significantly better growth than the addition of fresh A. filiculoides in sand. For fresh biomass, grain weights, Leaf area ratio (LAR) and relative growth rate (RGR), the performance of dried A. filiculoides was as good as that of the heated A. filiculoides. Productivity of wheat in the heated treatments increased significantly with increasing proportion of A. filiculoides added to sand, while in dry treatments there were no significant increases in productivity in the preliminary study. This supported the hypothesis that A. filiculoides, a notorious water weed can be put to agricultural use under dryland conditions in poor nutrient soils. Further investigations using dried A. filiculoides in sand and topsoil showed that the use of the same amounts of the dried fern made no significant short term impact on topsoil grown winter wheat but significantly improved the productivity of wheat in sand. Results showed that the addition of dried 20% Azolla to sand improved the soil fertility to levels equalling the quality of the control topsoil, but the addition of 80% Azolla to sand led to significantly greater wheat productivity than all other treatments. The addition of dried 20% Azolla (8.14 × 10^3 kg ha^(-1)) in sand produced as much wheat biomass as the addition of the recommended NPK fertiliser (30 kg N ha^(-1)) to sand. A comparison between the topsoil and sand-grown plants showed differences in flowering time but these had no effect on the final grain and above ground biomass.
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Water requirements and distribution of Ammophila arenaria and Scaevola plumieri on South African coastal dunes
- Authors: Peter, Craig Ingram
- Date: 2000
- Subjects: Scaevola plumieri , Sand dune plants , Sand dune planting , Plants -- Transpiration , Sandworts , Plant-water relationships , Evapotranspiration , Plants, Effect of heat on
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4219 , http://hdl.handle.net/10962/d1003788 , Scaevola plumieri , Sand dune plants , Sand dune planting , Plants -- Transpiration , Sandworts , Plant-water relationships , Evapotranspiration , Plants, Effect of heat on
- Description: Phenomenological models are presented which predicts transpiration rates (E) of individual leaves of Scaevola plumieri, an indigenous dune pioneer, and Ammophila arenaria, an exotic grass species introduced to stabilise mobile sand. In both cases E is predictably related to atmospheric vapour pressure deficit (VPD). VPD is calculated from measurements of ambient temperature and humidity, hence, where these two environmental variables are known, E can be calculated. Possible physiological reasons for the relationships of E to VPD in both species are discussed. Scaling from measurements of E at the leaf level to the canopy level is achieved by summing the leaf area of the canopy in question. E is predicted for the entire canopy leaf area by extrapolation to this larger leaf area. Predicted transpiration rates of individual shoot within the canopy were tested gravimetrically and shown to be accurate in the case of S. plumieri, but less so in the case of A. arenaria. Using this model, the amount of water used by a known area of sand dune is shown to be less than the rainfall input in the case of S. plumieri in wet and dry years. The water use of A. arenaria exceeds rainfall in the low-rainfall year of 1995, while in 1998 rainfall input is slightly higher than water extraction by the plants. Using a geographic information system (GIS), regional maps (surfaces) of transpiration were calculated from surfaces of mean monthly temperature and mean monthly relative humidity. Monthly surfaces of transpiration were subtracted from the monthly median rainfall to produce a surface of mean monthly water deficit. Areas of water surpluses along the coast correspond with the recorded distribution of both species in the seasons that the plants are most actively growing and reproducing. This suggests that unfavourable water availability during these two species growth periods limit their distributions along the coast. In addition to unfavourable water deficits, additional climatic variables that may be important in limiting the distribution of these two species were investigated using a discriminant function analysis.
- Full Text:
- Authors: Peter, Craig Ingram
- Date: 2000
- Subjects: Scaevola plumieri , Sand dune plants , Sand dune planting , Plants -- Transpiration , Sandworts , Plant-water relationships , Evapotranspiration , Plants, Effect of heat on
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4219 , http://hdl.handle.net/10962/d1003788 , Scaevola plumieri , Sand dune plants , Sand dune planting , Plants -- Transpiration , Sandworts , Plant-water relationships , Evapotranspiration , Plants, Effect of heat on
- Description: Phenomenological models are presented which predicts transpiration rates (E) of individual leaves of Scaevola plumieri, an indigenous dune pioneer, and Ammophila arenaria, an exotic grass species introduced to stabilise mobile sand. In both cases E is predictably related to atmospheric vapour pressure deficit (VPD). VPD is calculated from measurements of ambient temperature and humidity, hence, where these two environmental variables are known, E can be calculated. Possible physiological reasons for the relationships of E to VPD in both species are discussed. Scaling from measurements of E at the leaf level to the canopy level is achieved by summing the leaf area of the canopy in question. E is predicted for the entire canopy leaf area by extrapolation to this larger leaf area. Predicted transpiration rates of individual shoot within the canopy were tested gravimetrically and shown to be accurate in the case of S. plumieri, but less so in the case of A. arenaria. Using this model, the amount of water used by a known area of sand dune is shown to be less than the rainfall input in the case of S. plumieri in wet and dry years. The water use of A. arenaria exceeds rainfall in the low-rainfall year of 1995, while in 1998 rainfall input is slightly higher than water extraction by the plants. Using a geographic information system (GIS), regional maps (surfaces) of transpiration were calculated from surfaces of mean monthly temperature and mean monthly relative humidity. Monthly surfaces of transpiration were subtracted from the monthly median rainfall to produce a surface of mean monthly water deficit. Areas of water surpluses along the coast correspond with the recorded distribution of both species in the seasons that the plants are most actively growing and reproducing. This suggests that unfavourable water availability during these two species growth periods limit their distributions along the coast. In addition to unfavourable water deficits, additional climatic variables that may be important in limiting the distribution of these two species were investigated using a discriminant function analysis.
- Full Text: