title: Comparison of the efficiency of three methods of isolating fish DNA from environmental samples

authors: Banks, Jonathan C.

abstract: The Lake Ecosystem Restoration New Zealand (LERNZ) project was initiated to restore indigenous biodiversity in lakes in part by developing new pest fish management and control technologies. One objective of the management of pest fish was to develop genetic based assays to detect pest fish from environmental samples such as water, and to assess the feasibility of using these methods in the field.
This report compares three methods (the Piscitron II, Piscitron III and a pH adjusted method) of isolating DNA for two species of exotic fish from water samples. We compared DNA isolation efficiency using newly developed qPCR assays for koi carp, Cyprinus carpio and goldfish, Carassius auratus , and present the primer and hydrolysis probe sequences for the assays, and assay thermal conditions. Minimum levels of detectable fish biomass are reported for koi carp water collected from a 6800 L tank at ambient temperature containing a 1.6 kg koi carp. The koi carp and goldfish assays were also tested on Waikato River water collected next to Hammond Park, Hamilton, New Zealand, an area of river known to contain koi carp and goldfish.
We found that the Piscitron II and Piscitron III methods, which filtered water through glass fibre filters or glass wool aquarium filters, respectively, performed equally well in terms of DNA extraction but considerably better than the pH adjusted method. Minimum detectable amounts of koi carp DNA were calculated by producing serial dilutions of the tank water containing 2.35* 10⁻⁴ kg of koi carp/L. Because fish densities tend to be reported as kg/m² of lake surface, we converted our minimum detectable amount from a weight per volume to a weight per area by assuming an average lake depth of 1.5 m (a typical Waikato lake depth) which resulted in a minimum detectable amount of 0.35 kg carp/m² of water surface area. We also produced serial dilutions of DNA extracted from the koi carp tank and the minimum detectable amount of koi carp biomass was much lower at 0.0035 kg/m² which is less than the range of carp densities (0.005-0.0768 kg/m²) reported for Waikato lakes and rivers.

text: Comparison of the efficiency of three methods of isolating fish DNA from environmental samples The Lake Ecosystem Restoration New Zealand (LERNZ) project was initiated to restore indigenous biodiversity in lakes in part by developing new pest fish management and control technologies. One objective of the management of pest fish was to develop genetic based assays to detect pest fish from environmental samples such as water, and to assess the feasibility of using these methods in the field.
This report compares three methods (the Piscitron II, Piscitron III and a pH adjusted method) of isolating DNA for two species of exotic fish from water samples. We compared DNA isolation efficiency using newly developed qPCR assays for koi carp, Cyprinus carpio and goldfish, Carassius auratus , and present the primer and hydrolysis probe sequences for the assays, and assay thermal conditions. Minimum levels of detectable fish biomass are reported for koi carp water collected from a 6800 L tank at ambient temperature containing a 1.6 kg koi carp. The koi carp and goldfish assays were also tested on Waikato River water collected next to Hammond Park, Hamilton, New Zealand, an area of river known to contain koi carp and goldfish.
We found that the Piscitron II and Piscitron III methods, which filtered water through glass fibre filters or glass wool aquarium filters, respectively, performed equally well in terms of DNA extraction but considerably better than the pH adjusted method. Minimum detectable amounts of koi carp DNA were calculated by producing serial dilutions of the tank water containing 2.35* 10⁻⁴ kg of koi carp/L. Because fish densities tend to be reported as kg/m² of lake surface, we converted our minimum detectable amount from a weight per volume to a weight per area by assuming an average lake depth of 1.5 m (a typical Waikato lake depth) which resulted in a minimum detectable amount of 0.35 kg carp/m² of water surface area. We also produced serial dilutions of DNA extracted from the koi carp tank and the minimum detectable amount of koi carp biomass was much lower at 0.0035 kg/m² which is less than the range of carp densities (0.005-0.0768 kg/m²) reported for Waikato lakes and rivers.

year: 2014

title: Technological practice in early childhood as a dispositional milieu

authors: Carr, Margaret

subjects: early childhood education

abstract: This study investigated the learning of a group of four-year-olds as they worked in the construction area of a sessional early childhood setting. The researcher participated and observed as the children constructed with cardboard, rolled painted marbles around in a box to make Jackson Pollock style paintings, added collage and paint to pictures and constructions, and screen-printed cut-out figures and shapes of various kinds. These were described as technological practices, and five of them were compared as contexts for learning dispositions.
Although the literature had provided discussion and definition of dispositions in early childhood, the connection between the psychological notion of disposition and an historically or socioculturally framed dispositional milieu had not been researched. The study investigated empirical evidence for a transactional model of the interface between learning disposition and dispositional milieu, and learning dispositions were located in discourse, narrative and technological practice. The affordance of the technology and the tools played an important part in the transaction between individual and environment, in particular the technology's transparency, challenge, and accessibility. The five technological practices were analysed as complex sociocultural worlds in which multiple discourses jostled for privileged positions, children made decisions about whether discourse membership was for display or for exploration, they tackled difficulty with enthusiasm or avoided it in imaginative ways, and they engaged with others from a range of positions of power and responsibility. These sequences of events were described as learning narratives: historically and socioculturally co­constructed event structures about goals, challenge, and agency. A learning narrative was defined, identified, and investigated. 
Learning dispositions have been defined but not specified for early childhood. This study suggests that dispositions can be interpreted as parts of a learning narrative, and for this study key dispositional domains were identified as privileged discourses, preferred responses to difficulty, and favoured patterns of responsibility. Four learning dispositions emerged from these domains: courage, mindfulness, perseverance, and responsibility. These learning dispositions find a parallel in the strands of the early childhood curriculum document for Aotearoa New Zealand, Te Whāriki. The researcher followed up the observations by interviewing the children for their viewpoint on preferred responses to difficulty using an interview that reflected their experiences back to them as a picture book. 
Although much of the learning appeared to be attached to the context, and the privileged meaning and intent for that context was a key element, the transactional nature of the learning revealed individual learning dispositions and narratives that were developing in the early years and that may be robust and enduring.

text: Technological practice in early childhood as a dispositional milieu This study investigated the learning of a group of four-year-olds as they worked in the construction area of a sessional early childhood setting. The researcher participated and observed as the children constructed with cardboard, rolled painted marbles around in a box to make Jackson Pollock style paintings, added collage and paint to pictures and constructions, and screen-printed cut-out figures and shapes of various kinds. These were described as technological practices, and five of them were compared as contexts for learning dispositions.
Although the literature had provided discussion and definition of dispositions in early childhood, the connection between the psychological notion of disposition and an historically or socioculturally framed dispositional milieu had not been researched. The study investigated empirical evidence for a transactional model of the interface between learning disposition and dispositional milieu, and learning dispositions were located in discourse, narrative and technological practice. The affordance of the technology and the tools played an important part in the transaction between individual and environment, in particular the technology's transparency, challenge, and accessibility. The five technological practices were analysed as complex sociocultural worlds in which multiple discourses jostled for privileged positions, children made decisions about whether discourse membership was for display or for exploration, they tackled difficulty with enthusiasm or avoided it in imaginative ways, and they engaged with others from a range of positions of power and responsibility. These sequences of events were described as learning narratives: historically and socioculturally co­constructed event structures about goals, challenge, and agency. A learning narrative was defined, identified, and investigated. 
Learning dispositions have been defined but not specified for early childhood. This study suggests that dispositions can be interpreted as parts of a learning narrative, and for this study key dispositional domains were identified as privileged discourses, preferred responses to difficulty, and favoured patterns of responsibility. Four learning dispositions emerged from these domains: courage, mindfulness, perseverance, and responsibility. These learning dispositions find a parallel in the strands of the early childhood curriculum document for Aotearoa New Zealand, Te Whāriki. The researcher followed up the observations by interviewing the children for their viewpoint on preferred responses to difficulty using an interview that reflected their experiences back to them as a picture book. 
Although much of the learning appeared to be attached to the context, and the privileged meaning and intent for that context was a key element, the transactional nature of the learning revealed individual learning dispositions and narratives that were developing in the early years and that may be robust and enduring.

year: 1997

title: Rejoinder

authors: Kelly, Martin

abstract: Our paper (Kelly et al., 1999) set out to have points recognized as ‘needing to be addressed’ on the agendas of institutions currently providing accounting education. We believe that many institutions are continuing to fail to respond to recognized problems. We are grateful to the Commentators for recognizing our purpose. They note that our paper:
1. raises some important issues which need to be urgently addressed (Commentator 1);
2. illuminates why it is dif  cult to implement change even when there is a consensus to do so (Commentator 2);
3. provides a very important theme which has been neglected by many accounting educators (Commentator 3).

text: Rejoinder Our paper (Kelly et al., 1999) set out to have points recognized as ‘needing to be addressed’ on the agendas of institutions currently providing accounting education. We believe that many institutions are continuing to fail to respond to recognized problems. We are grateful to the Commentators for recognizing our purpose. They note that our paper:
1. raises some important issues which need to be urgently addressed (Commentator 1);
2. illuminates why it is dif  cult to implement change even when there is a consensus to do so (Commentator 2);
3. provides a very important theme which has been neglected by many accounting educators (Commentator 3).

year: 2000