Microbial and extracellular polymeric substance dynamics in arid–zone temporary pan ecosystems
- Authors: Bute, Tafara Frank
- Date: 2023-03-29
- Subjects: Extracellular polymeric substances , Biofilms , Vernal pools , Microbiomes , Sediment–water interface
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422258 , vital:71925
- Description: Microbial communities of bacteria, viruses, algae, protozoans and fungi participate profoundly in aquatic systems, particularly in mediating processes such as primary production, decomposition, and biogeochemical cycles. In addition, microbiomes produce extracellular polymeric substances (EPS) which encompass a hydrated exopolymer mainly constituted of carbohydrates and proteins. The exopolymer aid proliferation and persistence of biofilms on their resident surfaces. There is however paucity of data on functional diversity of microbiomes in arid zone temporary wetlands with previous research having mainly focused on permanent systems in the northern hemisphere. In the face of ongoing climatic changes and anthropogenic threats to wetlands, it is imperative to assess the health status of aquatic systems in relation to microbial productivity dynamics. In this thesis, colorimetric methods and sequence–based metagenomics were conducted to quantify microbial EPS production and bacterial metagenome functions, respectively. This study was conducted in Khakhea–Bray region (North–West, South Africa) in June 2021 and January 2022 with a focus on evaluating microbial patterns of distribution between seasons (i.e., Dry and Wet) and varying depth i.e., deepest zones (Deep), intermediate depth (Mid) and shallowest regions (Edge). Additionally, potential relationships between EPS and either water content or organic matter content (OM content) were evaluated. In this study it was hypothesized that wet phases and deeper zones will have high EPS production and support more functions in comparison to shallowest regions and dry phases. Carbohydrates and proteins were quantified using the Dubois method and modified Lowry procedure, respectively. Carbohydrates generally occurred in higher proportions than proteins, suggesting that EPS found in these systems was largely diatom produced. The wet phases (wet season and inundation periods) supported more EPS production compared to the dry phases. The results of principal components analysis (PCA) and Spearman’s correlations suggested that EPS was highly correlated with sediment water content among other assessed variables. No significant associations were established between EPS and organic matter content. Spatial distribution of EPS demonstrated similar patterns between the deepest (Deep) and the intermediate depth zones (Mid) however the shallow regions (Edge) had significantly lower concentrations. Bacterial characterization was established by amplification of the 16S rRNA gene using illumina–sequencing protocol. Enzyme functions associated with biogeochemical pathways were predicted in PICRUSt2 bioinformatics pipeline. A total of 15 042 Unique Amplicon Sequence Variants (ASVs) were observed to be affiliated to 51 bacterial phyla and 1 127 genera. All top genera had commonality in heat tolerance. Firmicutes, dominated at phyla level with 59 % (mean ± sd, 19 ± 13 %) relative abundance followed by Actinobacteria and Proteobacteria both at 34 % (18 ± 7 %) and (18 ± 6 %), respectively. Microbial diversity matrices highlighted significant differences in beta diversity more than alpha diversity. Bacterial microbiomes were more distinct between seasons compared to within season, suggesting that functions were seasonally driven. These findings were supported by highest rates of denitrification, carbohydrate degradation and EPS production by core microbiomes in the wet season as compared to low rates of nitrogen mineralisation, carbon fixation and nitrification in the dry season. The present findings represent a first attempt in evaluating sequence–based metagenomics in semi–arid southern African temporary pan ecosystem. Both microbial EPS and bacterial functional potential were highly driven by water availability, with highest rates mainly associated with maximum inundation compared to dry states of pans. It can therefore be suggested that extended dry periods are threatening to microbially mediated processes in temporary wetlands, with implications to loss of biodiversity due to desiccation resulting in poor nutrient cycling. , Thesis (MSc) -- Faculty of Science, Zoology and Entomology, 2023
- Full Text:
- Date Issued: 2023-03-29
- Authors: Bute, Tafara Frank
- Date: 2023-03-29
- Subjects: Extracellular polymeric substances , Biofilms , Vernal pools , Microbiomes , Sediment–water interface
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/422258 , vital:71925
- Description: Microbial communities of bacteria, viruses, algae, protozoans and fungi participate profoundly in aquatic systems, particularly in mediating processes such as primary production, decomposition, and biogeochemical cycles. In addition, microbiomes produce extracellular polymeric substances (EPS) which encompass a hydrated exopolymer mainly constituted of carbohydrates and proteins. The exopolymer aid proliferation and persistence of biofilms on their resident surfaces. There is however paucity of data on functional diversity of microbiomes in arid zone temporary wetlands with previous research having mainly focused on permanent systems in the northern hemisphere. In the face of ongoing climatic changes and anthropogenic threats to wetlands, it is imperative to assess the health status of aquatic systems in relation to microbial productivity dynamics. In this thesis, colorimetric methods and sequence–based metagenomics were conducted to quantify microbial EPS production and bacterial metagenome functions, respectively. This study was conducted in Khakhea–Bray region (North–West, South Africa) in June 2021 and January 2022 with a focus on evaluating microbial patterns of distribution between seasons (i.e., Dry and Wet) and varying depth i.e., deepest zones (Deep), intermediate depth (Mid) and shallowest regions (Edge). Additionally, potential relationships between EPS and either water content or organic matter content (OM content) were evaluated. In this study it was hypothesized that wet phases and deeper zones will have high EPS production and support more functions in comparison to shallowest regions and dry phases. Carbohydrates and proteins were quantified using the Dubois method and modified Lowry procedure, respectively. Carbohydrates generally occurred in higher proportions than proteins, suggesting that EPS found in these systems was largely diatom produced. The wet phases (wet season and inundation periods) supported more EPS production compared to the dry phases. The results of principal components analysis (PCA) and Spearman’s correlations suggested that EPS was highly correlated with sediment water content among other assessed variables. No significant associations were established between EPS and organic matter content. Spatial distribution of EPS demonstrated similar patterns between the deepest (Deep) and the intermediate depth zones (Mid) however the shallow regions (Edge) had significantly lower concentrations. Bacterial characterization was established by amplification of the 16S rRNA gene using illumina–sequencing protocol. Enzyme functions associated with biogeochemical pathways were predicted in PICRUSt2 bioinformatics pipeline. A total of 15 042 Unique Amplicon Sequence Variants (ASVs) were observed to be affiliated to 51 bacterial phyla and 1 127 genera. All top genera had commonality in heat tolerance. Firmicutes, dominated at phyla level with 59 % (mean ± sd, 19 ± 13 %) relative abundance followed by Actinobacteria and Proteobacteria both at 34 % (18 ± 7 %) and (18 ± 6 %), respectively. Microbial diversity matrices highlighted significant differences in beta diversity more than alpha diversity. Bacterial microbiomes were more distinct between seasons compared to within season, suggesting that functions were seasonally driven. These findings were supported by highest rates of denitrification, carbohydrate degradation and EPS production by core microbiomes in the wet season as compared to low rates of nitrogen mineralisation, carbon fixation and nitrification in the dry season. The present findings represent a first attempt in evaluating sequence–based metagenomics in semi–arid southern African temporary pan ecosystem. Both microbial EPS and bacterial functional potential were highly driven by water availability, with highest rates mainly associated with maximum inundation compared to dry states of pans. It can therefore be suggested that extended dry periods are threatening to microbially mediated processes in temporary wetlands, with implications to loss of biodiversity due to desiccation resulting in poor nutrient cycling. , Thesis (MSc) -- Faculty of Science, Zoology and Entomology, 2023
- Full Text:
- Date Issued: 2023-03-29
Dual and targeted photodynamic therapy ablation of bacterial and cancer cells using phthalocyanines and porphyrins in the presence of carbon-based nanomaterials
- Authors: Openda, Yolande Ikala
- Date: 2022-10-14
- Subjects: Phthalocyanines , Porphyrins , Active oxygen , Biofilms , Breast Cancer Treatment , Nanostructured materials , Combination therapy , Photochemotherapy
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/365945 , vital:65804 , DOI https://doi.org/10.21504/10962//365946
- Description: Phthalocyanines (Pcs) and porphyrins bearing substituents that possess antibacterial/anticancer properties are used as photosensitizers (PS) for the first time in the work. For targeting specificity and improved photoactivity, the PSs were afterward functionalized with carbon nanomaterials such as graphene quantum dots (GQDs) and detonation nanodiamonds (DNDs) via covalent conjugation (amide or ester bonds) or by non-covalent conjugation (π-π stacking and electrostatic interactions). Furthermore, the PSs-DNDs nanoconjugates were conjugated to either chitosan-capped silver nanoparticles (CSAg) via amide bonds or to the bare silver nanoparticles (Ag NPs) using the silver- nitrogen affinity. The as-synthesized nanoconjugates were also fully characterized by spectroscopic and microscopic methods together with thermal analysis. The potential photocytotoxicity of the complexes alone and their nanoconjugates against S. aureus and/or E. coli planktonic and biofilm cultures has been evaluated in vitro. Compared to the non- quaternized PSs, the cationic analogs exhibited a higher photodynamic inactivation against the planktonic cells with log10 reduction values above 9 in the viable count using a concentration of ca. 1.25 μM following 30 min exposure to light (Light dose: 943 J/cm2 for Pcs and 250 mW/cm2 for porphyrins). Whereas, at a concentration of ca. 100 μM the cationic PSs showed complete eradication of biofilms upon 30 min exposure to light. As a result of conjugation to carbon-based nanomaterials and silver nanoparticles, the compounds proved to be more effective as they exhibited stronger antibacterial and anti-biofilm activities on the multi-drug resistant bacteria strains due to synergetic effect, compared to PSs alone. This suggests that the newly prepared nanohybrids (PS concentration ca. 100 μM) could be used as potential antimicrobial agents in the treatment of biofilm-related infections. The target nanoconjugates showed all the advantages of two different groups existing on a single entity. In light of the potential advantages of combined chemotherapy and photodynamic antimicrobial chemotherapy (PACT), this work reports for the first time the use of PACT-ciprofloxacin (CIP) dual therapy using selected indium quaternized PSs which showed higher photoactivity with complete eradication of both Gram-positive and Gram-negative bacteria biofilms at concentrations of 8 μM of PS versus 2 μg/mL of the antibiotic following 15 min irradiation time (light dose: 471 J/cm2 for Pcs and fluence: 250 mW/cm2 for porphyrins) on S. aureus. Whereas the total killing of E. coli was obtained when combining 8 or 16 μM of PS combined with 4 μg/mL of CIP. The combined treatment resulted in the complete eradication of the matured biofilms with the highest log10 reduction values of 7.05 and 7.20 on S. aureus and E. coli, respectively. Used as a model, positively charged dimethylamino-chalcone Pcs also exhibited interesting photodynamic therapy (PDT) activity against MCF-7 cancer cells giving IC50 values of 17.9 and 7.4 μM, respectively following 15 min irradiation. Additionally, the TD-B3LYP/LanL2DZ calculations were run on the dimethylaminophenyl- porphyrins to compare the singlet excitation energies of quaternized and non-quaternized porphyrins in vacuo. the study shows excellent agreement between time-dependent density- functional theory (TD-DFT) exciting energies and the experimental S1>S0 excitation energies. The small deviation observed between the calculated and experimental spectra arises from the solvent effect. The excitation energies observed in these UV-Vis spectra mostly originated from electron promotion between the highest occupied molecular orbital (HOMO) for the less intense band and the HOMO-1 for the most intense band of the ground states to the lower unoccupied molecular orbital (LUMO) of the excited states. , Thesis (PhD) -- Faculty of Science, Chemistry, 2022
- Full Text:
- Date Issued: 2022-10-14
- Authors: Openda, Yolande Ikala
- Date: 2022-10-14
- Subjects: Phthalocyanines , Porphyrins , Active oxygen , Biofilms , Breast Cancer Treatment , Nanostructured materials , Combination therapy , Photochemotherapy
- Language: English
- Type: Academic theses , Doctoral theses , text
- Identifier: http://hdl.handle.net/10962/365945 , vital:65804 , DOI https://doi.org/10.21504/10962//365946
- Description: Phthalocyanines (Pcs) and porphyrins bearing substituents that possess antibacterial/anticancer properties are used as photosensitizers (PS) for the first time in the work. For targeting specificity and improved photoactivity, the PSs were afterward functionalized with carbon nanomaterials such as graphene quantum dots (GQDs) and detonation nanodiamonds (DNDs) via covalent conjugation (amide or ester bonds) or by non-covalent conjugation (π-π stacking and electrostatic interactions). Furthermore, the PSs-DNDs nanoconjugates were conjugated to either chitosan-capped silver nanoparticles (CSAg) via amide bonds or to the bare silver nanoparticles (Ag NPs) using the silver- nitrogen affinity. The as-synthesized nanoconjugates were also fully characterized by spectroscopic and microscopic methods together with thermal analysis. The potential photocytotoxicity of the complexes alone and their nanoconjugates against S. aureus and/or E. coli planktonic and biofilm cultures has been evaluated in vitro. Compared to the non- quaternized PSs, the cationic analogs exhibited a higher photodynamic inactivation against the planktonic cells with log10 reduction values above 9 in the viable count using a concentration of ca. 1.25 μM following 30 min exposure to light (Light dose: 943 J/cm2 for Pcs and 250 mW/cm2 for porphyrins). Whereas, at a concentration of ca. 100 μM the cationic PSs showed complete eradication of biofilms upon 30 min exposure to light. As a result of conjugation to carbon-based nanomaterials and silver nanoparticles, the compounds proved to be more effective as they exhibited stronger antibacterial and anti-biofilm activities on the multi-drug resistant bacteria strains due to synergetic effect, compared to PSs alone. This suggests that the newly prepared nanohybrids (PS concentration ca. 100 μM) could be used as potential antimicrobial agents in the treatment of biofilm-related infections. The target nanoconjugates showed all the advantages of two different groups existing on a single entity. In light of the potential advantages of combined chemotherapy and photodynamic antimicrobial chemotherapy (PACT), this work reports for the first time the use of PACT-ciprofloxacin (CIP) dual therapy using selected indium quaternized PSs which showed higher photoactivity with complete eradication of both Gram-positive and Gram-negative bacteria biofilms at concentrations of 8 μM of PS versus 2 μg/mL of the antibiotic following 15 min irradiation time (light dose: 471 J/cm2 for Pcs and fluence: 250 mW/cm2 for porphyrins) on S. aureus. Whereas the total killing of E. coli was obtained when combining 8 or 16 μM of PS combined with 4 μg/mL of CIP. The combined treatment resulted in the complete eradication of the matured biofilms with the highest log10 reduction values of 7.05 and 7.20 on S. aureus and E. coli, respectively. Used as a model, positively charged dimethylamino-chalcone Pcs also exhibited interesting photodynamic therapy (PDT) activity against MCF-7 cancer cells giving IC50 values of 17.9 and 7.4 μM, respectively following 15 min irradiation. Additionally, the TD-B3LYP/LanL2DZ calculations were run on the dimethylaminophenyl- porphyrins to compare the singlet excitation energies of quaternized and non-quaternized porphyrins in vacuo. the study shows excellent agreement between time-dependent density- functional theory (TD-DFT) exciting energies and the experimental S1>S0 excitation energies. The small deviation observed between the calculated and experimental spectra arises from the solvent effect. The excitation energies observed in these UV-Vis spectra mostly originated from electron promotion between the highest occupied molecular orbital (HOMO) for the less intense band and the HOMO-1 for the most intense band of the ground states to the lower unoccupied molecular orbital (LUMO) of the excited states. , Thesis (PhD) -- Faculty of Science, Chemistry, 2022
- Full Text:
- Date Issued: 2022-10-14
Settlement and metamorphosis in the veliger larvae of the South African abalone Haliotis midae exposed to ambient grown biofilms treated with conspecific mucous
- Authors: Van Staden, Jefferson Luke
- Date: 2021-10-29
- Subjects: Haliotis midae , Biofilms , Haliotis midae Larvae Behavior , Haliotis midae Metamorphosis , Biosecurity , Aquaculture , Metamorphosis , Attachment mechanisms (Biology) , Mucous
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/192187 , vital:45203
- Description: The South African abalone, Haliotis midae, is a commercially important species of mollusc which contributes significantly to the value of the South African mariculture industry. One of the primary challenges experienced by abalone farmers is the consistent production of juvenile abalone (spat) in sufficient volumes to keep stocking farms and facilitate expansion of the industry. One of the key production bottlenecks of H. midae is achieving adequate levels of larval attachment and metamorphosis (settlement). The larvae of H. midae are settled on polycarbonate plates which have been pre-conditioned with biofilms in seawater which is pumped ashore onto farms. The seasonal variability in settlement success reported by hatchery managers in South Africa is hypothesised to be as a result of different diatom species compositions colonising the settlement plates at different times of the year, with settlement success being lowest during the winter months. The following study investigated whether the addition of conspecific mucous to biofilms could result in elevated settlement success, and whether there was potential for sterilisation of this mucous. A novel method of mucous application, spraying it onto the plates as opposed to pre-grazing, was tested in settlement assays and the trials revealed the following results: • The addition of H. midae mucous induced significantly more larvae to attach to settlement plates, when mucous was harvested around the spawning season. • Elevated attachment of larvae on mucous treated plates did not result in more post-larvae occupying the plates at the end of trials, and increased mortality is likely attributed to introduction of pathogens in conjunction with mucous. • No increase in the final proportion of settled larvae which had metamorphosed or the rate at which they metamorphosed was observed between mucous application treatments and biofilm only treatments. Subsequent trials assessed whether methods of mucous handling could reduce the biosecurity risk associated with mucous use, and so mucous was either UV irradiated or autoclaved. These trials revealed the following findings: • No difference in attachment was seen between any treatments, including the untreated mucous. This is contrary to the findings of our initial experiments and illustrates that the attachment-inducing properties within mucous may be seasonally expressed. ii • Numbers of observed larvae/post-larvae on plates applied with UV and autoclaved mucous where less stable than biofilms only, especially in the second trial, illustrating that mucous still presents a biosecurity risk even after undergoing these handling methods as it may act as a substrate on which pathogenic bacteria could colonise. , Thesis (MSc) -- Faculty of Science, Ichthyology and Fisheries Science, 2021
- Full Text:
- Date Issued: 2021-10-29
- Authors: Van Staden, Jefferson Luke
- Date: 2021-10-29
- Subjects: Haliotis midae , Biofilms , Haliotis midae Larvae Behavior , Haliotis midae Metamorphosis , Biosecurity , Aquaculture , Metamorphosis , Attachment mechanisms (Biology) , Mucous
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/192187 , vital:45203
- Description: The South African abalone, Haliotis midae, is a commercially important species of mollusc which contributes significantly to the value of the South African mariculture industry. One of the primary challenges experienced by abalone farmers is the consistent production of juvenile abalone (spat) in sufficient volumes to keep stocking farms and facilitate expansion of the industry. One of the key production bottlenecks of H. midae is achieving adequate levels of larval attachment and metamorphosis (settlement). The larvae of H. midae are settled on polycarbonate plates which have been pre-conditioned with biofilms in seawater which is pumped ashore onto farms. The seasonal variability in settlement success reported by hatchery managers in South Africa is hypothesised to be as a result of different diatom species compositions colonising the settlement plates at different times of the year, with settlement success being lowest during the winter months. The following study investigated whether the addition of conspecific mucous to biofilms could result in elevated settlement success, and whether there was potential for sterilisation of this mucous. A novel method of mucous application, spraying it onto the plates as opposed to pre-grazing, was tested in settlement assays and the trials revealed the following results: • The addition of H. midae mucous induced significantly more larvae to attach to settlement plates, when mucous was harvested around the spawning season. • Elevated attachment of larvae on mucous treated plates did not result in more post-larvae occupying the plates at the end of trials, and increased mortality is likely attributed to introduction of pathogens in conjunction with mucous. • No increase in the final proportion of settled larvae which had metamorphosed or the rate at which they metamorphosed was observed between mucous application treatments and biofilm only treatments. Subsequent trials assessed whether methods of mucous handling could reduce the biosecurity risk associated with mucous use, and so mucous was either UV irradiated or autoclaved. These trials revealed the following findings: • No difference in attachment was seen between any treatments, including the untreated mucous. This is contrary to the findings of our initial experiments and illustrates that the attachment-inducing properties within mucous may be seasonally expressed. ii • Numbers of observed larvae/post-larvae on plates applied with UV and autoclaved mucous where less stable than biofilms only, especially in the second trial, illustrating that mucous still presents a biosecurity risk even after undergoing these handling methods as it may act as a substrate on which pathogenic bacteria could colonise. , Thesis (MSc) -- Faculty of Science, Ichthyology and Fisheries Science, 2021
- Full Text:
- Date Issued: 2021-10-29
The biology and molecular ecology of floating sulphur biofilms
- Authors: Bowker, Michelle Louise
- Date: 2002
- Subjects: Biofilms , Microbial ecology , Sulfur
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4056 , http://hdl.handle.net/10962/d1004117 , Biofilms , Microbial ecology , Sulfur
- Description: Floating sulphur biofilms have been observed to occur on sulphate-containing natural systems and waste stabilization ponds. It has been postulated that these biofilms form on the surface of the water because sulphate reducing bacteria present in the bottom layers of the water body reduce sulphate to sulphide which then diffuses upwards and is oxidized under the correct redox conditions to sulphur by sulphide oxidizing bacteria. Very little information exists on these complex floating systems and in order to study them further, model systems were designed. The Baffle Reactor was successfully used to cultivate floating sulphur biofilms. Conditions within the reactor could be closely scrutinized in the laboratory and it was found that sulphate levels decreased, sulphide levels increased and that sulphur was produced over a period of 2 weeks. The success of this system led to it being scaled-up and currently a method to harvest sulphur from the biofilm is under development. It is thought that biofilms are highly complex, heterogeneous structures with different bacteria distributed in different layers. Preliminary work suggested that bacteria were differentially distributed along nutrient and oxygen gradients within the biofilm. Biofilms are very thin structures and therefore difficult to study and Gradient systems were developed in an attempt to spatially separate the biofilm species into functional layers. Gradient Tubes were designed; these provided a gradient of high-sulphide, low oxygen conditions to high-oxygen, low-sulphide conditions. Bacteria were observed to grow in different layers of these systems. The Gradient Tubes could be sectioned and the chemical characteristics of each section as well as the species present could be determined. Silicon Tubular Bioreactors were also developed and these were very efficient at producing large amounts of sulphur under strictly controlled redox conditions. Microscopy and molecular methods including the amplification of a section of Ribosomal Ribonucleic acid by Polymerase Chain Reaction were used in an attempt to characterize the populations present in these biofilm systems. Denaturing Gradient Gel Electrophoresis was used to create band profiles of the populations; individual bands were excised from the gels and sequenced. Identified species included Ectothiorhodospira sp., Dethiosulfovibrio russensis, Pseudomonas geniculata, Thiobacillus baregensis and Halothiobacillus kellyi.
- Full Text:
- Date Issued: 2002
- Authors: Bowker, Michelle Louise
- Date: 2002
- Subjects: Biofilms , Microbial ecology , Sulfur
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4056 , http://hdl.handle.net/10962/d1004117 , Biofilms , Microbial ecology , Sulfur
- Description: Floating sulphur biofilms have been observed to occur on sulphate-containing natural systems and waste stabilization ponds. It has been postulated that these biofilms form on the surface of the water because sulphate reducing bacteria present in the bottom layers of the water body reduce sulphate to sulphide which then diffuses upwards and is oxidized under the correct redox conditions to sulphur by sulphide oxidizing bacteria. Very little information exists on these complex floating systems and in order to study them further, model systems were designed. The Baffle Reactor was successfully used to cultivate floating sulphur biofilms. Conditions within the reactor could be closely scrutinized in the laboratory and it was found that sulphate levels decreased, sulphide levels increased and that sulphur was produced over a period of 2 weeks. The success of this system led to it being scaled-up and currently a method to harvest sulphur from the biofilm is under development. It is thought that biofilms are highly complex, heterogeneous structures with different bacteria distributed in different layers. Preliminary work suggested that bacteria were differentially distributed along nutrient and oxygen gradients within the biofilm. Biofilms are very thin structures and therefore difficult to study and Gradient systems were developed in an attempt to spatially separate the biofilm species into functional layers. Gradient Tubes were designed; these provided a gradient of high-sulphide, low oxygen conditions to high-oxygen, low-sulphide conditions. Bacteria were observed to grow in different layers of these systems. The Gradient Tubes could be sectioned and the chemical characteristics of each section as well as the species present could be determined. Silicon Tubular Bioreactors were also developed and these were very efficient at producing large amounts of sulphur under strictly controlled redox conditions. Microscopy and molecular methods including the amplification of a section of Ribosomal Ribonucleic acid by Polymerase Chain Reaction were used in an attempt to characterize the populations present in these biofilm systems. Denaturing Gradient Gel Electrophoresis was used to create band profiles of the populations; individual bands were excised from the gels and sequenced. Identified species included Ectothiorhodospira sp., Dethiosulfovibrio russensis, Pseudomonas geniculata, Thiobacillus baregensis and Halothiobacillus kellyi.
- Full Text:
- Date Issued: 2002
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:
- Date Issued: 2000
- 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:
- Date Issued: 2000
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