title: The preparation of fluorescent fingerprinting powders using 8-hydroxyquinoline compounds and magnetite

authors: O'Shea, Benjamin

subjects: Fingerprint

abstract: Fluorescent fingerprinting powders were prepared by using rotary evaporation to coat magnetite powder with varying amounts of either tris(8-hydroxyquinolinato)aluminium (Alq₃), tris(8-hydroxyquinolinato)gallium (Gaq₃), bis(8-hydroxyquinolinato)magnesium (Mgq₂) or bis(8-hydroxyquinolinato)zinc (Znq₂). The performance of each fingerprinting powder was tested on various surfaces under visible light and under UV excitation. The efficacy of powder application using a magnetic brush was compared to that of standard brushes. The effects of varying the particle sizes of the fingerprinting powders made with Alq₃ and Znq₂ were explored. The stability of each fluorescent compound under standard conditions was tested.
The synthesis of sodium tetrakis(8-hydroxyquinolinato)boron (NaBq₄) was attempted twice, but failed both times. The product acquired was suspected to be Na₄(C₉H₆NO)₄(H₂O)₈ (Na₄q₄(H₂O)₈) instead. Crystals of Na₄q₄(H₂O)₈ were prepared using a modified literature method. The Na₄q₄(H₂O)₈ crystals and the products of the attempted syntheses were characterised by powder X-ray crystal diffraction, inductively coupled plasma mass spectrometry, electrospray ionisation mass spectrometry, nuclear magnetic resonance spectroscopy, UV-Vis absorption, and melting point measurements. Fingerprint powders were prepared using the products of the attempted NaBq₄ syntheses and tested in the same manner as the Alq₃, Gaq₃, Znq₂, and Mgq₂ fingerprint powders.

text: The preparation of fluorescent fingerprinting powders using 8-hydroxyquinoline compounds and magnetite Fluorescent fingerprinting powders were prepared by using rotary evaporation to coat magnetite powder with varying amounts of either tris(8-hydroxyquinolinato)aluminium (Alq₃), tris(8-hydroxyquinolinato)gallium (Gaq₃), bis(8-hydroxyquinolinato)magnesium (Mgq₂) or bis(8-hydroxyquinolinato)zinc (Znq₂). The performance of each fingerprinting powder was tested on various surfaces under visible light and under UV excitation. The efficacy of powder application using a magnetic brush was compared to that of standard brushes. The effects of varying the particle sizes of the fingerprinting powders made with Alq₃ and Znq₂ were explored. The stability of each fluorescent compound under standard conditions was tested.
The synthesis of sodium tetrakis(8-hydroxyquinolinato)boron (NaBq₄) was attempted twice, but failed both times. The product acquired was suspected to be Na₄(C₉H₆NO)₄(H₂O)₈ (Na₄q₄(H₂O)₈) instead. Crystals of Na₄q₄(H₂O)₈ were prepared using a modified literature method. The Na₄q₄(H₂O)₈ crystals and the products of the attempted syntheses were characterised by powder X-ray crystal diffraction, inductively coupled plasma mass spectrometry, electrospray ionisation mass spectrometry, nuclear magnetic resonance spectroscopy, UV-Vis absorption, and melting point measurements. Fingerprint powders were prepared using the products of the attempted NaBq₄ syntheses and tested in the same manner as the Alq₃, Gaq₃, Znq₂, and Mgq₂ fingerprint powders.

year: 2018

title: Induced reproduction in Pātiki (Rhombosolea leporina), a novel endemic aquaculture candidate

authors: Ellis-Smith, Brooke Georgia

subjects: Aquaculture

abstract: The rapid global population growth combined with the decline of wild-capture fisheries has resulted in the global expansion of aquaculture. New Zealand’s aquaculture industry remains largely underdeveloped and as such, industry growth would benefit from species diversification. The endemic yellowbelly flounder (Rhombosolea leporina) is a promising candidate species for New Zealand aquaculture due to its high market value, fast growth rate and the global demand for flatfishes. At present, very limited knowledge exists regarding the reproductive biology of this species, and notably the control of reproduction in captivity. Successful finfish aquaculture requires a robust understanding of reproduction for effective broodstock management. This thesis investigates induced ovulation in yellowbelly flounder using a gonadotropin releasing hormone analogue (GnRHa). The study pairs physiology and genetics to advance the current knowledge of reproduction in this species.
GnRHa was administrated to induce ovulation in yellowbelly flounder, over a 28-day period. One of three treatments were assigned to 27 female yellowbelly flounder (n=9): 50 µg/kg GnRHa, 100 µg/kg GnRHa, or a sham treatment of 50 µg/kg Ringer’s solution. Histological and fresh oocyte analyses were carried out and batch fecundity, fertilization and ovulation rates were assessed. Reproductive dysfunction experienced by yellowbelly flounder in this study was through incomplete germinal vesicle migration and a failure to enter hydration. Ovulation was only observed in GnRHa treated fish and the ovulatory periodicity appeared to be approximately 72 hours in fish that ovulated more than once. This study confirms that yellowbelly flounder are multiple group synchronous batch spawners. Although fewer fish than expected ovulated (four), significantly more GnRHa treated fish completed final oocyte maturation and entered hydration compared to the controls. Furthermore, fecundity, fertilization rates and egg viability were high, with fertilization >80% achieved in some initial ovulations. In contrast, eggs obtained from fish that had ovulated for a second time, were characterised by low fecundity and fertilization rates. Nonetheless, this data indicates GnRHa can successfully induce ovulation at dosages of at least 50 µg/kg without adversely affecting egg quality in at least the first batch. Further refinement is essential to optimize a protocol for the reliable induction of reproduction in this species. Low incidences of ovulation may be a factor of dosage or inappropriate ovarian maturity at the time of hormone administration. This highlights the need to confirm oocyte maturity immediately prior to GnRHa treatment. To aid future work, a classification index for the identification of ovarian developmental stage based on fresh oocyte morphology was developed for yellowbelly flounder. 
To further advance future research into yellowbelly flounder reproduction, brain, pituitary and gonad transcriptomic libraries were generated for female yellowbelly flounder using RNA-seq. Protein sequences of eight reproduction related genes were successfully identified and characterised, which were StAR, Gpr54, Lhβ, Fshβ, Fshr, Cyp19a1a, Cyp19a1b and Hsd17b1. Conserved functional regions, phylogenetic analysis and homology with sequences from other Pleuronectiformes confirmed the identities of these genes and indicated that their functional roles are likely conserved. Characterisation of these genes provides highly valuable information for future quantitative assessment of gene expression throughout the yellowbelly flounder reproductive axis. 
The results presented in this thesis will help to inform future work to control and optimize reproduction in captive yellowbelly flounder. In addition, the genetic data will enable molecular tools to be developed, including quantitative PCR primers as well as the potential for recombinant gonadotropins to develop enzyme-linked immunosorbent assays. These tools will be highly valuable to accurately assess the effects of GnRHa treatment on reproduction and gene expression under differing culture conditions. This study indicates that successful ovulation and good quality eggs may be achieved in yellowbelly flounder using GnRHa treatment.

text: Induced reproduction in Pātiki (Rhombosolea leporina), a novel endemic aquaculture candidate The rapid global population growth combined with the decline of wild-capture fisheries has resulted in the global expansion of aquaculture. New Zealand’s aquaculture industry remains largely underdeveloped and as such, industry growth would benefit from species diversification. The endemic yellowbelly flounder (Rhombosolea leporina) is a promising candidate species for New Zealand aquaculture due to its high market value, fast growth rate and the global demand for flatfishes. At present, very limited knowledge exists regarding the reproductive biology of this species, and notably the control of reproduction in captivity. Successful finfish aquaculture requires a robust understanding of reproduction for effective broodstock management. This thesis investigates induced ovulation in yellowbelly flounder using a gonadotropin releasing hormone analogue (GnRHa). The study pairs physiology and genetics to advance the current knowledge of reproduction in this species.
GnRHa was administrated to induce ovulation in yellowbelly flounder, over a 28-day period. One of three treatments were assigned to 27 female yellowbelly flounder (n=9): 50 µg/kg GnRHa, 100 µg/kg GnRHa, or a sham treatment of 50 µg/kg Ringer’s solution. Histological and fresh oocyte analyses were carried out and batch fecundity, fertilization and ovulation rates were assessed. Reproductive dysfunction experienced by yellowbelly flounder in this study was through incomplete germinal vesicle migration and a failure to enter hydration. Ovulation was only observed in GnRHa treated fish and the ovulatory periodicity appeared to be approximately 72 hours in fish that ovulated more than once. This study confirms that yellowbelly flounder are multiple group synchronous batch spawners. Although fewer fish than expected ovulated (four), significantly more GnRHa treated fish completed final oocyte maturation and entered hydration compared to the controls. Furthermore, fecundity, fertilization rates and egg viability were high, with fertilization >80% achieved in some initial ovulations. In contrast, eggs obtained from fish that had ovulated for a second time, were characterised by low fecundity and fertilization rates. Nonetheless, this data indicates GnRHa can successfully induce ovulation at dosages of at least 50 µg/kg without adversely affecting egg quality in at least the first batch. Further refinement is essential to optimize a protocol for the reliable induction of reproduction in this species. Low incidences of ovulation may be a factor of dosage or inappropriate ovarian maturity at the time of hormone administration. This highlights the need to confirm oocyte maturity immediately prior to GnRHa treatment. To aid future work, a classification index for the identification of ovarian developmental stage based on fresh oocyte morphology was developed for yellowbelly flounder. 
To further advance future research into yellowbelly flounder reproduction, brain, pituitary and gonad transcriptomic libraries were generated for female yellowbelly flounder using RNA-seq. Protein sequences of eight reproduction related genes were successfully identified and characterised, which were StAR, Gpr54, Lhβ, Fshβ, Fshr, Cyp19a1a, Cyp19a1b and Hsd17b1. Conserved functional regions, phylogenetic analysis and homology with sequences from other Pleuronectiformes confirmed the identities of these genes and indicated that their functional roles are likely conserved. Characterisation of these genes provides highly valuable information for future quantitative assessment of gene expression throughout the yellowbelly flounder reproductive axis. 
The results presented in this thesis will help to inform future work to control and optimize reproduction in captive yellowbelly flounder. In addition, the genetic data will enable molecular tools to be developed, including quantitative PCR primers as well as the potential for recombinant gonadotropins to develop enzyme-linked immunosorbent assays. These tools will be highly valuable to accurately assess the effects of GnRHa treatment on reproduction and gene expression under differing culture conditions. This study indicates that successful ovulation and good quality eggs may be achieved in yellowbelly flounder using GnRHa treatment.

year: 2022

title: Overview of the Kukutaaruhe Gully restoration initiative: Part 1

authors: Elliot Noe, Elizabeth

abstract: Kukutaaruhe Gully is a minor branch of Hamilton’s extensive gully system, and runs from the corner of Bankwood and Clarkin Road, behind Fairfield College, connecting with Donny Park in the north and out into the Waikato River. The Kukutaaruhe Education Trust was established to support the implementation of the Fairfield Project, which is a partnership between Ngati Wairere, local community and Fairfield College. The project’s goals include the establishment of an ecological and environmental education centre on the approximately 12 ha of land behind Fairfield College and restoration of the adjacent Kukutaaruhe Gully. In 2019, the University of Waikato’s Environmental Research Institute was contracted by the Kukutaaruhe Education Trust to create an Overview of the Kukutaaruhe Gully restoration initiative. This is one in a three-part series created together under one contract:
• Part 1 of 3 is the first document presented here, an Overview of the Kukutaaruhe Gully Restoration Initiative
• Part 2 of 3, the Operational Forest Restoration plan for Kukutaaruhe Gully
• Part 3 of 3 is an Ecological Restoration Plan Template.

text: Overview of the Kukutaaruhe Gully restoration initiative: Part 1 Kukutaaruhe Gully is a minor branch of Hamilton’s extensive gully system, and runs from the corner of Bankwood and Clarkin Road, behind Fairfield College, connecting with Donny Park in the north and out into the Waikato River. The Kukutaaruhe Education Trust was established to support the implementation of the Fairfield Project, which is a partnership between Ngati Wairere, local community and Fairfield College. The project’s goals include the establishment of an ecological and environmental education centre on the approximately 12 ha of land behind Fairfield College and restoration of the adjacent Kukutaaruhe Gully. In 2019, the University of Waikato’s Environmental Research Institute was contracted by the Kukutaaruhe Education Trust to create an Overview of the Kukutaaruhe Gully restoration initiative. This is one in a three-part series created together under one contract:
• Part 1 of 3 is the first document presented here, an Overview of the Kukutaaruhe Gully Restoration Initiative
• Part 2 of 3, the Operational Forest Restoration plan for Kukutaaruhe Gully
• Part 3 of 3 is an Ecological Restoration Plan Template.

year: 2020-10