title: ¹⁵N stable isotope probing of pulp and paper wastewaters

authors: Addison, Sarah Louise

subjects: Stable isotope probing

abstract: Stable isotope probing (SIP) is an established technique that can be applied to identify the metabolically active micro-organisms within a microbial population. The SIP method utilises an isotopically-labelled substrate and PCR techniques to discern the members of a microbial community that incorporate the isotope into their DNA or RNA. The current literature gap around using ¹⁵N isotopes with RNA-SIP offers real potential and advantages for targeting and identifying active members from mixed communities involved in global biogeochemical nitrogen cycling.

This study specifically investigated whether nitrogen based compounds ncan be used as substrates in RNA-SIP methodologies and whether they can in turn be used to probe mixed community environments known to be actively fixing nitrogen. The nitrogen-limited systems targeted represented an ideal opportunity to assess the suitability of ¹⁵N-RNA-SIP approaches due to their known high nitrogen fixation rates. Identifying these nitrogen-fixing bacteria could provide a better representation analysis of the community, leading to an improved prediction on how to manage and optimise the treatment performance of target waste systems and to exploit the unique bioconversion properties of these types of organisms.

Initially, the project undertook methodological proof of concept by using a soluble nitrogen source, ¹⁵NH₄Cl, to label the RNA of Novosphingobium nitrogenifigens and a mixed microbial community. Successful separation of the ¹⁴N- (control) and ¹⁵N-RNA was achieved for both pure and mixed communities using isopycnic caesium trifluoroacetate (CsTFA) gradients in an ultracentrifuge. The usefulness of this technique to identify active diazotrophs in real environmental samples was tested using a nitrogen-fixing community from a pulp and paper wastewater treatment system. After growing the mixed culture with ¹⁵N₂ as the sole nitrogen source, the labelled RNA was extracted and fractionated using isopycnic centrifugation in CsTFA gradients.

The community composition of the active nitrogen-fixing community in the ¹⁵N₂ enriched fraction was analysed by establishing a 16S rRNA gene clone library containing over 200 members. These were analysed by comparison with published sequences and by phylogenetic analysis.

It was found that the more isotopic label substrate incorporated, the further the buoyant density (BD) separation between ¹⁵N- and ¹⁴N-RNA. Novosphingobium nitrogenifigens gave an average BD shift of 0.03 + 0.004 g ml⁻¹ (95.0 atom % ¹⁵N) with ¹⁵NH₄Cl. For mixed communities the average BD shift was 0.02 + 0.004 g ml⁻¹ (80.0 atom % ¹⁵N) with 15NH4Cl and 0.013 + 0.002 g ml⁻¹ (32.6 atom % ¹⁵N) when using ¹⁵N₂. Clone library analysis of 16S rRNA genes present in the enriched ¹⁵N-RNA fraction of the mixed community was shown to consist of a diverse population of bacteria as indicated by a Shannon Weaver index value of >2.8. Three dominant genera (Aeromonas, Pseudomonas and Bacillus) were identified by comparison with published sequences and phylogenetic analysis. Many other groups not known as archetypal nitrogen-fixing bacteria were also identified, demonstrating that ¹⁵N₂-RNA-SIP provides a useful tool for the identification of important and previously unknown contributors to nitrogen fixation in a range of environments.

Overall, this project has established that nitrogen based RNA-SIP is a powerful tool that can be used successfully and reproducibly with both pure and complex mixed microbial communities to study active diazotrophs in environmental samples.

text: ¹⁵N stable isotope probing of pulp and paper wastewaters Stable isotope probing (SIP) is an established technique that can be applied to identify the metabolically active micro-organisms within a microbial population. The SIP method utilises an isotopically-labelled substrate and PCR techniques to discern the members of a microbial community that incorporate the isotope into their DNA or RNA. The current literature gap around using ¹⁵N isotopes with RNA-SIP offers real potential and advantages for targeting and identifying active members from mixed communities involved in global biogeochemical nitrogen cycling.

This study specifically investigated whether nitrogen based compounds ncan be used as substrates in RNA-SIP methodologies and whether they can in turn be used to probe mixed community environments known to be actively fixing nitrogen. The nitrogen-limited systems targeted represented an ideal opportunity to assess the suitability of ¹⁵N-RNA-SIP approaches due to their known high nitrogen fixation rates. Identifying these nitrogen-fixing bacteria could provide a better representation analysis of the community, leading to an improved prediction on how to manage and optimise the treatment performance of target waste systems and to exploit the unique bioconversion properties of these types of organisms.

Initially, the project undertook methodological proof of concept by using a soluble nitrogen source, ¹⁵NH₄Cl, to label the RNA of Novosphingobium nitrogenifigens and a mixed microbial community. Successful separation of the ¹⁴N- (control) and ¹⁵N-RNA was achieved for both pure and mixed communities using isopycnic caesium trifluoroacetate (CsTFA) gradients in an ultracentrifuge. The usefulness of this technique to identify active diazotrophs in real environmental samples was tested using a nitrogen-fixing community from a pulp and paper wastewater treatment system. After growing the mixed culture with ¹⁵N₂ as the sole nitrogen source, the labelled RNA was extracted and fractionated using isopycnic centrifugation in CsTFA gradients.

The community composition of the active nitrogen-fixing community in the ¹⁵N₂ enriched fraction was analysed by establishing a 16S rRNA gene clone library containing over 200 members. These were analysed by comparison with published sequences and by phylogenetic analysis.

It was found that the more isotopic label substrate incorporated, the further the buoyant density (BD) separation between ¹⁵N- and ¹⁴N-RNA. Novosphingobium nitrogenifigens gave an average BD shift of 0.03 + 0.004 g ml⁻¹ (95.0 atom % ¹⁵N) with ¹⁵NH₄Cl. For mixed communities the average BD shift was 0.02 + 0.004 g ml⁻¹ (80.0 atom % ¹⁵N) with 15NH4Cl and 0.013 + 0.002 g ml⁻¹ (32.6 atom % ¹⁵N) when using ¹⁵N₂. Clone library analysis of 16S rRNA genes present in the enriched ¹⁵N-RNA fraction of the mixed community was shown to consist of a diverse population of bacteria as indicated by a Shannon Weaver index value of >2.8. Three dominant genera (Aeromonas, Pseudomonas and Bacillus) were identified by comparison with published sequences and phylogenetic analysis. Many other groups not known as archetypal nitrogen-fixing bacteria were also identified, demonstrating that ¹⁵N₂-RNA-SIP provides a useful tool for the identification of important and previously unknown contributors to nitrogen fixation in a range of environments.

Overall, this project has established that nitrogen based RNA-SIP is a powerful tool that can be used successfully and reproducibly with both pure and complex mixed microbial communities to study active diazotrophs in environmental samples.

year: 2008

title: Promoting engagement and learning in first year university studies: The role of personalisation

authors: Stanley, Peter

subjects: university studies

abstract: Student engagement in higher education can be conceptualised as involving three components: students’ social needs and circumstances, the cognitive characteristics of academic studies, and the prevailing institutional ethos or philosophy that specifies the relationships that students have with learning and knowledge. This paper reports on an investigation into student engagement in a first-year human development course at the University of Waikato at Tauranga, New Zealand where the teaching staff has a commitment to relating learning to individual experiences. Information from an end-of-course survey indicates that a philosophy of personalisation promotes learning engagement. Students reported that they were required to think a lot or a great deal, that they put time into the course assessments, and that they valued the human development course itself.

text: Promoting engagement and learning in first year university studies: The role of personalisation Student engagement in higher education can be conceptualised as involving three components: students’ social needs and circumstances, the cognitive characteristics of academic studies, and the prevailing institutional ethos or philosophy that specifies the relationships that students have with learning and knowledge. This paper reports on an investigation into student engagement in a first-year human development course at the University of Waikato at Tauranga, New Zealand where the teaching staff has a commitment to relating learning to individual experiences. Information from an end-of-course survey indicates that a philosophy of personalisation promotes learning engagement. Students reported that they were required to think a lot or a great deal, that they put time into the course assessments, and that they valued the human development course itself.

year: 2011

title: Mormonism and the white possessive: Moving critical indigenous studies theory into the religious realm

authors: Simon, Hemopereki

abstract: This essay responds to the pressing need to pursue topics related to Mormonism from a research perspective that is at once critical, culturally affirming, and grounded in Indigenous views. The goal here is to provide a theoretical framework for Critical Indigenous Studies perspectives to emerge in Mormon and Lamanite Studies. I therefore submit the following questions as a contribution to and critique of Lamanite Studies within the broader fields of Mormon, Cultural, and Religious Studies:
1. Which theoretical frame(s) from Critical Indigenous 
Studies is/are best suited to address Lamanite and Mormon issues? 
2. How might we reframe current Critical Indigenous 
Studies Theory to undertake research on Mormon and 
Western religious topics? 

To address these questions, the article first outlines its 
positionality as a basis for understanding Simon’s approach to the research. The bulk of the research here focuses on describing Lamanitism in terms of its relationship to racism and whiteness. The article therefore continues with a description of the figure of 
Hagoth in The Book of Mormon, with a particular emphasis on their relationship to Te Moana-Nui-ā-Kiwa. From there the article outlines Unger’s theorem of contextuality and considers how Lamanitism might be considered an “artificial context” from a Critical Indigenous Studies perspective. A discussion of the Doctrine of Discovery, Manifest Destiny, and Moana Jackson’s idea of the culture of colonization is followed by a description of the “Racial Contract” and its speculative extension to Indigenous adherents of the Mormon Church. The article then explicates important and interrelated tikanga values from Te Ao Māori such as tapu and mana. Lastly, the article identifies the “white possessive” as a key doctrine for Critical Indigenous Studies and undertakes a reworking of the doctrine for its application to the Mormon Church. More specifically, I transmute the notion “white patriarchal sovereignty” into “white patriarchal salvation” in order to tailor it to critical Indigenous discussions of the Church. 
In my conclusion I attempt to answer the question – What is to be done? I assert that the Church is in conflict with the cultural revival of Te Ao Māori and the idea of “original instructions.” The Church’s claim to salvific truth and thinking is contradicted by my very identity and culturally informed ontology. The ensuing discussion confirms that as a colonizing entity the Church operates parallel to the government as a co-invading force in Indigenous lands. The idea of a Lamanite people as Polynesians, which includes Māori is called into question. I provide a continuum model to explain Indigenous realities in relation to religiosity and decolonization and suggest some topics as case studies for further investigation. The true purpose of this article is to facilitate the integration of critical Indigenous with Mormon and/or Lamanite Studies to bring about much-needed dialogue, discussion, and debate on the place of indigeneity in The Church of Jesus Christ of Latter-Day Saints.

text: Mormonism and the white possessive: Moving critical indigenous studies theory into the religious realm This essay responds to the pressing need to pursue topics related to Mormonism from a research perspective that is at once critical, culturally affirming, and grounded in Indigenous views. The goal here is to provide a theoretical framework for Critical Indigenous Studies perspectives to emerge in Mormon and Lamanite Studies. I therefore submit the following questions as a contribution to and critique of Lamanite Studies within the broader fields of Mormon, Cultural, and Religious Studies:
1. Which theoretical frame(s) from Critical Indigenous 
Studies is/are best suited to address Lamanite and Mormon issues? 
2. How might we reframe current Critical Indigenous 
Studies Theory to undertake research on Mormon and 
Western religious topics? 

To address these questions, the article first outlines its 
positionality as a basis for understanding Simon’s approach to the research. The bulk of the research here focuses on describing Lamanitism in terms of its relationship to racism and whiteness. The article therefore continues with a description of the figure of 
Hagoth in The Book of Mormon, with a particular emphasis on their relationship to Te Moana-Nui-ā-Kiwa. From there the article outlines Unger’s theorem of contextuality and considers how Lamanitism might be considered an “artificial context” from a Critical Indigenous Studies perspective. A discussion of the Doctrine of Discovery, Manifest Destiny, and Moana Jackson’s idea of the culture of colonization is followed by a description of the “Racial Contract” and its speculative extension to Indigenous adherents of the Mormon Church. The article then explicates important and interrelated tikanga values from Te Ao Māori such as tapu and mana. Lastly, the article identifies the “white possessive” as a key doctrine for Critical Indigenous Studies and undertakes a reworking of the doctrine for its application to the Mormon Church. More specifically, I transmute the notion “white patriarchal sovereignty” into “white patriarchal salvation” in order to tailor it to critical Indigenous discussions of the Church. 
In my conclusion I attempt to answer the question – What is to be done? I assert that the Church is in conflict with the cultural revival of Te Ao Māori and the idea of “original instructions.” The Church’s claim to salvific truth and thinking is contradicted by my very identity and culturally informed ontology. The ensuing discussion confirms that as a colonizing entity the Church operates parallel to the government as a co-invading force in Indigenous lands. The idea of a Lamanite people as Polynesians, which includes Māori is called into question. I provide a continuum model to explain Indigenous realities in relation to religiosity and decolonization and suggest some topics as case studies for further investigation. The true purpose of this article is to facilitate the integration of critical Indigenous with Mormon and/or Lamanite Studies to bring about much-needed dialogue, discussion, and debate on the place of indigeneity in The Church of Jesus Christ of Latter-Day Saints.

year: 2022