Aspects of the population ecology, habitat use and behaviour of the endangered Knysna seahorse (Hippocampus capensis Boulenger, 1900) in a residential marina estate, Knysna, South Africa: implications for conservation
- Authors: Claassens, Louw
- Date: 2018
- Subjects: Endangered species -- South Africa -- Knysna , Sea horses -- Behavior -- South Africa -- Knysna , Sea horses -- Habitat -- South Africa -- Knysna , Sea horses -- Ecology -- South Africa -- Knysna
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/54789 , vital:26616
- Description: The Knysna seahorse Hippocampus capensis is South Africa’s only endemic seahorse species, and is found in only three adjacent estuaries along the southern coast. The conservation of this endangered species is important on a national and international level. This study presents the first research on this species within the Knysna estuary since 2001 and specifically focuses on aspects of its ecology within a residential marina estate (Thesen Islands Marina). The physico-chemical and habitat features of the marina were described and the population ecology, habitat use, and behaviour of the Knysna seahorse were investigated. Physico-chemical conditions within the western section of the marina, characterised by high water current velocities, were similar to that of the adjacent estuary. The eastern section of the marina was characterised by lower water current velocities and higher turbidity. Four major habitat types were identified within the marina canals: (I) artificial Reno mattress (wire baskets filled with rocks); (II) Codium tenue beds; (III) mixed vegetation on sediment; and (IV) barren canal floor. Seahorse densities within the marina were significantly higher compared to densities found historically within the estuary. Highest seahorse densities were specifically found within the artificial Reno mattress structures and within the western section of the marina. Seahorse density varied spatially and temporally and the type of habitat was an important predictor for seahorse occurrence. An experimental investigation found that H. capensis chooses artificial Reno mattress habitat over Zostera capensis when given a choice. GoPro cameras were used successfully to investigate daytime seahorse behaviour within the Reno mattress habitat. Seahorses were more active during the morning, spent most of their time (> 80 %) feeding, and morning courting behaviour for this species were confirmed. However, during the summer holiday period (mid-December to mid-January) few seahorses were observed on camera, which suggests that the increase in motor boat activity and the related increase in noise had a negative effect on H. capensis feeding and courting behaviour. The marina development, and in particular the Reno mattresses, created a new habitat for this endangered species within the Knysna estuary. In addition to the protection and restoration of natural habitats in which H. capensis is found, the conservation potential of artificial structures such as Reno mattresses should be realised.
- Full Text:
- Date Issued: 2018
- Authors: Claassens, Louw
- Date: 2018
- Subjects: Endangered species -- South Africa -- Knysna , Sea horses -- Behavior -- South Africa -- Knysna , Sea horses -- Habitat -- South Africa -- Knysna , Sea horses -- Ecology -- South Africa -- Knysna
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/54789 , vital:26616
- Description: The Knysna seahorse Hippocampus capensis is South Africa’s only endemic seahorse species, and is found in only three adjacent estuaries along the southern coast. The conservation of this endangered species is important on a national and international level. This study presents the first research on this species within the Knysna estuary since 2001 and specifically focuses on aspects of its ecology within a residential marina estate (Thesen Islands Marina). The physico-chemical and habitat features of the marina were described and the population ecology, habitat use, and behaviour of the Knysna seahorse were investigated. Physico-chemical conditions within the western section of the marina, characterised by high water current velocities, were similar to that of the adjacent estuary. The eastern section of the marina was characterised by lower water current velocities and higher turbidity. Four major habitat types were identified within the marina canals: (I) artificial Reno mattress (wire baskets filled with rocks); (II) Codium tenue beds; (III) mixed vegetation on sediment; and (IV) barren canal floor. Seahorse densities within the marina were significantly higher compared to densities found historically within the estuary. Highest seahorse densities were specifically found within the artificial Reno mattress structures and within the western section of the marina. Seahorse density varied spatially and temporally and the type of habitat was an important predictor for seahorse occurrence. An experimental investigation found that H. capensis chooses artificial Reno mattress habitat over Zostera capensis when given a choice. GoPro cameras were used successfully to investigate daytime seahorse behaviour within the Reno mattress habitat. Seahorses were more active during the morning, spent most of their time (> 80 %) feeding, and morning courting behaviour for this species were confirmed. However, during the summer holiday period (mid-December to mid-January) few seahorses were observed on camera, which suggests that the increase in motor boat activity and the related increase in noise had a negative effect on H. capensis feeding and courting behaviour. The marina development, and in particular the Reno mattresses, created a new habitat for this endangered species within the Knysna estuary. In addition to the protection and restoration of natural habitats in which H. capensis is found, the conservation potential of artificial structures such as Reno mattresses should be realised.
- Full Text:
- Date Issued: 2018
The investigation of type-specific features of the copper coordinating AA9 proteins and their effect on the interaction with crystalline cellulose using molecular dynamics studies
- Authors: Moses, Vuyani
- Date: 2018
- Subjects: Copper proteins , Cellulose , Molecular dynamics , Cellulose -- Biodegradation , Bioinformatics
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/58327 , vital:27230
- Description: AA9 proteins are metallo-enzymes which are crucial for the early stages of cellulose degradation. AA9 proteins have been suggested to cleave glycosidic bonds linking cellulose through the use of their Cu2+ coordinating active site. AA9 proteins possess different regioselectivities depending on the resulting cleavage they form and as result, are grouped accordingly. Type 1 AA9 proteins cleave the C1 carbon of cellulose while Type 2 AA9 proteins cleave the C4 carbon and Type 3 AA9 proteins cleave either C1 or C4 carbons. The steric congestion of the AA9 active site has been proposed to be a contributor to the observed regioselectivity. As such, a bioinformatics characterisation of type-specific sequence and structural features was performed. Initially AA9 protein sequences were obtained from the Pfam database and multiple sequence alignment was performed. The sequences were phylogenetically characterised and sequences were grouped into their respective types and sub-groups were identified. A selection analysis was performed on AA9 LPMO types to determine the selective pressure acting on AA9 protein residues. Motif discovery was then performed to identify conserved sequence motifs in AA9 proteins. Once type-specific sequence features were identified structural mapping was performed to assess possible effects on substrate interaction. Physicochemical property analysis was also performed to assess biochemical differences between AA9 LPMO types. Molecular dynamics (MD) simulations were then employed to dynamically assess the consequences of the discovered type-specific features on AA9-cellulose interaction. Due to the absence of AA9 specific force field parameters MD simulations were not readily applicable. As a result, Potential Energy Surface (PES) scans were performed to evaluate the force field parameters for the AA9 active site using the PM6 semi empirical approach and least squares fitting. A Type 1 AA9 active site was constructed from the crystal structure 4B5Q, encompassing only the Cu2+ coordinating residues, the Cu2+ ion and two water residues. Due to the similarity in AA9 active sites, the Type force field parameters were validated on all three AA9 LPMO types. Two MD simulations for each AA9 LPMO types were conducted using two separate Lennard-Jones parameter sets. Once completed, the MD trajectories were analysed for various features including the RMSD, RMSF, radius of gyration, coordination during simulation, hydrogen bonding, secondary structure conservation and overall protein movement. Force field parameters were successfully evaluated and validated for AA9 proteins. MD simulations of AA9 proteins were able to reveal the presence of unique type-specific binding modes of AA9 active sites to cellulose. These binding modes were characterised by the presence of unique type-specific loops which were present in Type 2 and 3 AA9 proteins but not in Type 1 AA9 proteins. The loops were found to result in steric congestion that affects how the Cu2+ ion interacts with cellulose. As a result, Cu2+ binding to cellulose was observed for Type 1 and not Type 2 and 3 AA9 proteins. In this study force field parameters have been evaluated for the Type 1 active site of AA9 proteins and this parameters were evaluated on all three types and binding. Future work will focus on identifying the nature of the reactive oxygen species and performing QM/MM calculations to elucidate the reactive mechanism of all three AA9 LPMO types.
- Full Text:
- Date Issued: 2018
- Authors: Moses, Vuyani
- Date: 2018
- Subjects: Copper proteins , Cellulose , Molecular dynamics , Cellulose -- Biodegradation , Bioinformatics
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/58327 , vital:27230
- Description: AA9 proteins are metallo-enzymes which are crucial for the early stages of cellulose degradation. AA9 proteins have been suggested to cleave glycosidic bonds linking cellulose through the use of their Cu2+ coordinating active site. AA9 proteins possess different regioselectivities depending on the resulting cleavage they form and as result, are grouped accordingly. Type 1 AA9 proteins cleave the C1 carbon of cellulose while Type 2 AA9 proteins cleave the C4 carbon and Type 3 AA9 proteins cleave either C1 or C4 carbons. The steric congestion of the AA9 active site has been proposed to be a contributor to the observed regioselectivity. As such, a bioinformatics characterisation of type-specific sequence and structural features was performed. Initially AA9 protein sequences were obtained from the Pfam database and multiple sequence alignment was performed. The sequences were phylogenetically characterised and sequences were grouped into their respective types and sub-groups were identified. A selection analysis was performed on AA9 LPMO types to determine the selective pressure acting on AA9 protein residues. Motif discovery was then performed to identify conserved sequence motifs in AA9 proteins. Once type-specific sequence features were identified structural mapping was performed to assess possible effects on substrate interaction. Physicochemical property analysis was also performed to assess biochemical differences between AA9 LPMO types. Molecular dynamics (MD) simulations were then employed to dynamically assess the consequences of the discovered type-specific features on AA9-cellulose interaction. Due to the absence of AA9 specific force field parameters MD simulations were not readily applicable. As a result, Potential Energy Surface (PES) scans were performed to evaluate the force field parameters for the AA9 active site using the PM6 semi empirical approach and least squares fitting. A Type 1 AA9 active site was constructed from the crystal structure 4B5Q, encompassing only the Cu2+ coordinating residues, the Cu2+ ion and two water residues. Due to the similarity in AA9 active sites, the Type force field parameters were validated on all three AA9 LPMO types. Two MD simulations for each AA9 LPMO types were conducted using two separate Lennard-Jones parameter sets. Once completed, the MD trajectories were analysed for various features including the RMSD, RMSF, radius of gyration, coordination during simulation, hydrogen bonding, secondary structure conservation and overall protein movement. Force field parameters were successfully evaluated and validated for AA9 proteins. MD simulations of AA9 proteins were able to reveal the presence of unique type-specific binding modes of AA9 active sites to cellulose. These binding modes were characterised by the presence of unique type-specific loops which were present in Type 2 and 3 AA9 proteins but not in Type 1 AA9 proteins. The loops were found to result in steric congestion that affects how the Cu2+ ion interacts with cellulose. As a result, Cu2+ binding to cellulose was observed for Type 1 and not Type 2 and 3 AA9 proteins. In this study force field parameters have been evaluated for the Type 1 active site of AA9 proteins and this parameters were evaluated on all three types and binding. Future work will focus on identifying the nature of the reactive oxygen species and performing QM/MM calculations to elucidate the reactive mechanism of all three AA9 LPMO types.
- Full Text:
- Date Issued: 2018
Transcription factor binding specificity and occupancy : elucidation, modelling and evaluation
- Authors: Kibet, Caleb Kipkurui
- Date: 2017
- Subjects: Transcription factors , Transcription factors -- Data processing , Motif Assessment and Ranking Suite
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:21185 , http://hdl.handle.net/10962/6838
- Description: The major contributions of this thesis are addressing the need for an objective quality evaluation of a transcription factor binding model, demonstrating the value of the tools developed to this end and elucidating how in vitro and in vivo information can be utilized to improve TF binding specificity models. Accurate elucidation of TF binding specificity remains an ongoing challenge in gene regulatory research. Several in vitro and in vivo experimental techniques have been developed followed by a proliferation of algorithms, and ultimately, the binding models. This increase led to a choice problem for the end users: which tools to use, and which is the most accurate model for a given TF? Therefore, the first section of this thesis investigates the motif assessment problem: how scoring functions, choice and processing of benchmark data, and statistics used in evaluation affect motif ranking. This analysis revealed that TF motif quality assessment requires a systematic comparative analysis, and that scoring functions used have a TF-specific effect on motif ranking. These results advised the design of a Motif Assessment and Ranking Suite MARS, supported by PBM and ChIP-seq benchmark data and an extensive collection of PWM motifs. MARS implements consistency, enrichment, and scoring and classification-based motif evaluation algorithms. Transcription factor binding is also influenced and determined by contextual factors: chromatin accessibility, competition or cooperation with other TFs, cell line or condition specificity, binding locality (e.g. proximity to transcription start sites) and the shape of the binding site (DNA-shape). In vitro techniques do not capture such context; therefore, this thesis also combines PBM and DNase-seq data using a comparative k-mer enrichment approach that compares open chromatin with genome-wide prevalence, achieving a modest performance improvement when benchmarked on ChIP-seq data. Finally, since statistical and probabilistic methods cannot capture all the information that determine binding, a machine learning approach (XGBooost) was implemented to investigate how the features contribute to TF specificity and occupancy. This combinatorial approach improves the predictive ability of TF specificity models with the most predictive feature being chromatin accessibility, while the DNA-shape and conservation information all significantly improve on the baseline model of k-mer and DNase data. The results and the tools introduced in this thesis are useful for systematic comparative analysis (via MARS) and a combinatorial approach to modelling TF binding specificity, including appropriate feature engineering practices for machine learning modelling.
- Full Text:
- Date Issued: 2017
- Authors: Kibet, Caleb Kipkurui
- Date: 2017
- Subjects: Transcription factors , Transcription factors -- Data processing , Motif Assessment and Ranking Suite
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:21185 , http://hdl.handle.net/10962/6838
- Description: The major contributions of this thesis are addressing the need for an objective quality evaluation of a transcription factor binding model, demonstrating the value of the tools developed to this end and elucidating how in vitro and in vivo information can be utilized to improve TF binding specificity models. Accurate elucidation of TF binding specificity remains an ongoing challenge in gene regulatory research. Several in vitro and in vivo experimental techniques have been developed followed by a proliferation of algorithms, and ultimately, the binding models. This increase led to a choice problem for the end users: which tools to use, and which is the most accurate model for a given TF? Therefore, the first section of this thesis investigates the motif assessment problem: how scoring functions, choice and processing of benchmark data, and statistics used in evaluation affect motif ranking. This analysis revealed that TF motif quality assessment requires a systematic comparative analysis, and that scoring functions used have a TF-specific effect on motif ranking. These results advised the design of a Motif Assessment and Ranking Suite MARS, supported by PBM and ChIP-seq benchmark data and an extensive collection of PWM motifs. MARS implements consistency, enrichment, and scoring and classification-based motif evaluation algorithms. Transcription factor binding is also influenced and determined by contextual factors: chromatin accessibility, competition or cooperation with other TFs, cell line or condition specificity, binding locality (e.g. proximity to transcription start sites) and the shape of the binding site (DNA-shape). In vitro techniques do not capture such context; therefore, this thesis also combines PBM and DNase-seq data using a comparative k-mer enrichment approach that compares open chromatin with genome-wide prevalence, achieving a modest performance improvement when benchmarked on ChIP-seq data. Finally, since statistical and probabilistic methods cannot capture all the information that determine binding, a machine learning approach (XGBooost) was implemented to investigate how the features contribute to TF specificity and occupancy. This combinatorial approach improves the predictive ability of TF specificity models with the most predictive feature being chromatin accessibility, while the DNA-shape and conservation information all significantly improve on the baseline model of k-mer and DNase data. The results and the tools introduced in this thesis are useful for systematic comparative analysis (via MARS) and a combinatorial approach to modelling TF binding specificity, including appropriate feature engineering practices for machine learning modelling.
- Full Text:
- Date Issued: 2017
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