title: Optimisation of eDNA replication and detection distance in Aotearoa New Zealand lotic systems

authors: Smith, Joshua

abstract: The use of environmental DNA (eDNA) is a relatively new tool in New Zealand, and its increased use worldwide in the last decade has seen a surge in its utility and application. The information gaps in its use and effectiveness in the field of environmental science are fast being explored and answered. However, many questions remain unanswered, particularly in an Aotearoa New Zealand context. The aim of this thesis is to extend the knowledge needed for effective and efficient use of this new and exciting tool in lotic Aotearoa New Zealand conditions.

National standardisation of ecological sampling protocols between different agencies in Aotearoa New Zealand has historically been difficult to attain, creating challenges for combining datasets for national scale analyses. The introduction of new methods for biological monitoring, such as environmental DNA (eDNA), presents an opportunity to standardise aquatic sampling protocols prior to widespread adoption. The objective of this study (chapter two) was to optimise eDNA sample replication for the consistent characterisation of freshwater fish and macroinvertebrate communities in flowing waters, and ultimately, to inform the development of robust national monitoring standards. A comparison of field replication and extraction methods (pooling of preservation buffer) was also trialled as part of this high replication (n=16) eDNA study to assess any potential benefits in measuring species richness and reducing processing costs alongside replication optimisation. This involved two ‘syringe’ sampling methods (‘standard’ and ‘boosted’, eight each) conducted across 54 riverine sites throughout the country. No significant difference was found for species richness between the standardised (eight replicates) or boosted (16 replicates composited to eight) eDNA methods for fish and macroinvertebrates. Results indicated that six replicates were needed to consistently detect 89.5% of fish species likely to be present using field-based syringe eDNA sampling and preservation. However, an altitudinal species richness effect was observed for fish. For macroinvertebrates, six replicates were required to identify 86% of taxa identified to the NEMS (National Environmental Monitoring Standards) level used for the Macroinvertebrate Community Index (MCI: usually genera) while eight replicates were required to detect 89% of NEMS taxa. For fish and macroinvertebrate biodiversity, this study suggests that six replicates are a reasonable trade-off between effective community characterisation and cost in Aotearoa New Zealand lotic systems.

Understanding the spatial limits and the way in which eDNA behaves in lotic systems is important to understand when designing eDNA sampling programmes and interpreting results. Questions around biodiversity detection at spatiotemporal scales in relation to stream order and the influence of tributaries remain. We sought to extend our knowledge of longitudinal eDNA dispersal and detectability in wadable lotic systems by using live caged fish and a naturally occurring habitat specific fish species. An additional unexpected external molecular source was also included in the analysis. An eDNA collection method comparison between active and passive sampling techniques was also trialled alongside caged experiments in the second half of this study. A correlation between stream discharge and the distance in which eDNA travels downstream is reported. An initial drop in eDNA concentration from caged fish over the first kilometre downstream is followed by a peak 4 km downstream, is suggested to be reflective of plume dynamics. Tributary-specific fish species at low biomass and their associated eDNA signal is largely diluted and not detectable directly downstream of confluences, though there was some evidence that if sampled further downstream in the mainstem, tributary signals may be detected. The caged fish trials showed how far an eDNA signal can travel, while results from naturally occurring longitudinal signals confirm that eDNA metabarcoding can measure biodiversity at spatiotemporal scales. Active sampling in general outperformed passive sampling. Issues around field contamination and sampling strategies based on results from this study are also discussed.

Overall, this thesis provides insights and potential guidance in the use of eDNA in lotic Aotearoa New Zealand systems. It highlights the importance of eDNA field replication in the measurement of biodiversity for fish and macroinvertebrates and some terrestrial species. It helps interpret the ecology of eDNA in distance and structure. This information may assist in future eDNA sampling strategies and monitoring.

text: Optimisation of eDNA replication and detection distance in Aotearoa New Zealand lotic systems The use of environmental DNA (eDNA) is a relatively new tool in New Zealand, and its increased use worldwide in the last decade has seen a surge in its utility and application. The information gaps in its use and effectiveness in the field of environmental science are fast being explored and answered. However, many questions remain unanswered, particularly in an Aotearoa New Zealand context. The aim of this thesis is to extend the knowledge needed for effective and efficient use of this new and exciting tool in lotic Aotearoa New Zealand conditions.

National standardisation of ecological sampling protocols between different agencies in Aotearoa New Zealand has historically been difficult to attain, creating challenges for combining datasets for national scale analyses. The introduction of new methods for biological monitoring, such as environmental DNA (eDNA), presents an opportunity to standardise aquatic sampling protocols prior to widespread adoption. The objective of this study (chapter two) was to optimise eDNA sample replication for the consistent characterisation of freshwater fish and macroinvertebrate communities in flowing waters, and ultimately, to inform the development of robust national monitoring standards. A comparison of field replication and extraction methods (pooling of preservation buffer) was also trialled as part of this high replication (n=16) eDNA study to assess any potential benefits in measuring species richness and reducing processing costs alongside replication optimisation. This involved two ‘syringe’ sampling methods (‘standard’ and ‘boosted’, eight each) conducted across 54 riverine sites throughout the country. No significant difference was found for species richness between the standardised (eight replicates) or boosted (16 replicates composited to eight) eDNA methods for fish and macroinvertebrates. Results indicated that six replicates were needed to consistently detect 89.5% of fish species likely to be present using field-based syringe eDNA sampling and preservation. However, an altitudinal species richness effect was observed for fish. For macroinvertebrates, six replicates were required to identify 86% of taxa identified to the NEMS (National Environmental Monitoring Standards) level used for the Macroinvertebrate Community Index (MCI: usually genera) while eight replicates were required to detect 89% of NEMS taxa. For fish and macroinvertebrate biodiversity, this study suggests that six replicates are a reasonable trade-off between effective community characterisation and cost in Aotearoa New Zealand lotic systems.

Understanding the spatial limits and the way in which eDNA behaves in lotic systems is important to understand when designing eDNA sampling programmes and interpreting results. Questions around biodiversity detection at spatiotemporal scales in relation to stream order and the influence of tributaries remain. We sought to extend our knowledge of longitudinal eDNA dispersal and detectability in wadable lotic systems by using live caged fish and a naturally occurring habitat specific fish species. An additional unexpected external molecular source was also included in the analysis. An eDNA collection method comparison between active and passive sampling techniques was also trialled alongside caged experiments in the second half of this study. A correlation between stream discharge and the distance in which eDNA travels downstream is reported. An initial drop in eDNA concentration from caged fish over the first kilometre downstream is followed by a peak 4 km downstream, is suggested to be reflective of plume dynamics. Tributary-specific fish species at low biomass and their associated eDNA signal is largely diluted and not detectable directly downstream of confluences, though there was some evidence that if sampled further downstream in the mainstem, tributary signals may be detected. The caged fish trials showed how far an eDNA signal can travel, while results from naturally occurring longitudinal signals confirm that eDNA metabarcoding can measure biodiversity at spatiotemporal scales. Active sampling in general outperformed passive sampling. Issues around field contamination and sampling strategies based on results from this study are also discussed.

Overall, this thesis provides insights and potential guidance in the use of eDNA in lotic Aotearoa New Zealand systems. It highlights the importance of eDNA field replication in the measurement of biodiversity for fish and macroinvertebrates and some terrestrial species. It helps interpret the ecology of eDNA in distance and structure. This information may assist in future eDNA sampling strategies and monitoring.

year: 2024

title: Electromyographic assessment of pullover exercise variations and javelin throw on nine upper-body muscles

authors: Washif, Jad Adrian

abstract: Exercise choice is one important variable for attaining athletic training aims. Pullover exercises are integrated into resistance training programs of javelin throwers to develop event-specific strength. We examined the electromyography (EMG) signals during four pullover exercise variations: conventional (CONV), straight-arm (STR), bent-arm (BENT), and conventional on unstable surface (UNS); and javelin throwing (JAVT) using an 800 g standard implement. Generally, BENT elicited the highest peak activity of all muscles (22-89% MVC), except for the rectus abdominus which was highest during UNS (106%). In contrast, CONV tended to produce the lowest peak muscle activation (12-77%), except for the pectoral and middle deltoid muscles. It was noteworthy that despite being the same exercise, performance on the unstable surface elicited higher peak muscle activity than CONV, except from the pectoralis muscles. Peak muscle activation in JAVT ranged from 29 (posterior deltoid) to 124% (latissimus dorsi), with the relatively high peaks in the deltoids and latissimus dorsi not being matched during the pullover exercises. Surface EMG of five national male javelin throwers were recorded from nine muscles on the throwing-arm side: upper trapezius; anterior, middle, and posterior deltoid; triceps brachii; clavicular and sternal portions of pectoralis major; latissimus dorsi, and rectus abdominus. In a randomised fashion, all throwers performed six repetitions of each pullover variation using a free-weight curl-barbell against an external load at 85% of 1-repetition maximum, while EMG signals were recorded during both the concentric and eccentric phases of the movement and subsequently normalized to a maximal voluntary contraction (MVC). The throwers then performed three maximal javelin throws and the one with the greatest throwing velocity was used for analysis. It has been noted that pullover exercise emphasize chest muscle activation, i.e. pectoralis major (Marchetti & Uchida, 2011). Here, the level of activation depended on pullover variation, and BENT elicited the highest overall peak muscle activity. Strength and conditioning specialists should be aware of the impact of different pullover variations on muscle activation levels, and note that additional exercises to target the musculature of the shoulder and upper back should be incorporated in the training of javelin throwers.

text: Electromyographic assessment of pullover exercise variations and javelin throw on nine upper-body muscles Exercise choice is one important variable for attaining athletic training aims. Pullover exercises are integrated into resistance training programs of javelin throwers to develop event-specific strength. We examined the electromyography (EMG) signals during four pullover exercise variations: conventional (CONV), straight-arm (STR), bent-arm (BENT), and conventional on unstable surface (UNS); and javelin throwing (JAVT) using an 800 g standard implement. Generally, BENT elicited the highest peak activity of all muscles (22-89% MVC), except for the rectus abdominus which was highest during UNS (106%). In contrast, CONV tended to produce the lowest peak muscle activation (12-77%), except for the pectoral and middle deltoid muscles. It was noteworthy that despite being the same exercise, performance on the unstable surface elicited higher peak muscle activity than CONV, except from the pectoralis muscles. Peak muscle activation in JAVT ranged from 29 (posterior deltoid) to 124% (latissimus dorsi), with the relatively high peaks in the deltoids and latissimus dorsi not being matched during the pullover exercises. Surface EMG of five national male javelin throwers were recorded from nine muscles on the throwing-arm side: upper trapezius; anterior, middle, and posterior deltoid; triceps brachii; clavicular and sternal portions of pectoralis major; latissimus dorsi, and rectus abdominus. In a randomised fashion, all throwers performed six repetitions of each pullover variation using a free-weight curl-barbell against an external load at 85% of 1-repetition maximum, while EMG signals were recorded during both the concentric and eccentric phases of the movement and subsequently normalized to a maximal voluntary contraction (MVC). The throwers then performed three maximal javelin throws and the one with the greatest throwing velocity was used for analysis. It has been noted that pullover exercise emphasize chest muscle activation, i.e. pectoralis major (Marchetti & Uchida, 2011). Here, the level of activation depended on pullover variation, and BENT elicited the highest overall peak muscle activity. Strength and conditioning specialists should be aware of the impact of different pullover variations on muscle activation levels, and note that additional exercises to target the musculature of the shoulder and upper back should be incorporated in the training of javelin throwers.

year: 2016

title: Intellectual Property, Mātauranga Māori

authors: Sterling, Rogena

abstract: This literature review has been conducted to consider the various national legislation and international agreements that comprise New Zealand’s Intellectual Property Rights (IP) regime. It will evaluate if and how such legislation and agreements protect and enable Māori IP rights and interests with respect to Māori data, genomic data and mātauranga Māori. The review also identifies some mechanisms that might also enhance Māori control of these types of data. The Westminster approach of legislation in New Zealand and its approach to IP protection based on Copyrights, Patents and Trade Marks are juxtaposed against traditional Maori approaches of communally held ancestral knowledge (mātauranga) passed down through generations (whanaungatanga) based on guardianship and protection (kaitiakitanga) and the self-determination of use of such knowledge (rangatiratanga). Attempting to align tikanga concepts to the Westmionster model of law is challenging as the two share completely different notions of ownership and responsibility. Expectations of protection, to prevent misappropriation and commercialisation by non- Māori of mātauranga Māori and Māori data, extend beyond the parameters of existing IP law, creates a similar disjunct. Genomic Research generates data, some of which can be protected by IP, however researchers working with genetic/genomic data from taonga species have often failed to acknowledge the non-IP interests of Maori. As a result, Maori have taken it upon themselves to advocate for their rights to data through Māori data sovereignty discourse as well as create guidelines for culturally appropriate genomic research with explicit references to data security and management (e.g. Te Mata Ira, Te Nohonga Kaitiaki). Other extra-legal options, such as Biocultural Labelling to alert users where particular data has Māori rights and/or interests, are emerging to maintain create durable provenance data and connect next users of data with the responsible Indigenous communities. Though the intellectual property regime in New Zealand may provide some protections, there are still significant areas where the legal system does not provide sufficient protections for Māori data, taonga species and mātauranga. The UN Declaration on the Rights of Indigenous Peoples provides a framework of international support for Indigenous rights, but local government and other home-grown mechanisms are important to enable Māori governance of data. Local approaches Māori Data Sovereignty and Māori Data Governance afford Maori the opportunity to be directly involved as kaitiaki of their mātauranga and assert rangatiratanga over data and its use.

text: Intellectual Property, Mātauranga Māori This literature review has been conducted to consider the various national legislation and international agreements that comprise New Zealand’s Intellectual Property Rights (IP) regime. It will evaluate if and how such legislation and agreements protect and enable Māori IP rights and interests with respect to Māori data, genomic data and mātauranga Māori. The review also identifies some mechanisms that might also enhance Māori control of these types of data. The Westminster approach of legislation in New Zealand and its approach to IP protection based on Copyrights, Patents and Trade Marks are juxtaposed against traditional Maori approaches of communally held ancestral knowledge (mātauranga) passed down through generations (whanaungatanga) based on guardianship and protection (kaitiakitanga) and the self-determination of use of such knowledge (rangatiratanga). Attempting to align tikanga concepts to the Westmionster model of law is challenging as the two share completely different notions of ownership and responsibility. Expectations of protection, to prevent misappropriation and commercialisation by non- Māori of mātauranga Māori and Māori data, extend beyond the parameters of existing IP law, creates a similar disjunct. Genomic Research generates data, some of which can be protected by IP, however researchers working with genetic/genomic data from taonga species have often failed to acknowledge the non-IP interests of Maori. As a result, Maori have taken it upon themselves to advocate for their rights to data through Māori data sovereignty discourse as well as create guidelines for culturally appropriate genomic research with explicit references to data security and management (e.g. Te Mata Ira, Te Nohonga Kaitiaki). Other extra-legal options, such as Biocultural Labelling to alert users where particular data has Māori rights and/or interests, are emerging to maintain create durable provenance data and connect next users of data with the responsible Indigenous communities. Though the intellectual property regime in New Zealand may provide some protections, there are still significant areas where the legal system does not provide sufficient protections for Māori data, taonga species and mātauranga. The UN Declaration on the Rights of Indigenous Peoples provides a framework of international support for Indigenous rights, but local government and other home-grown mechanisms are important to enable Māori governance of data. Local approaches Māori Data Sovereignty and Māori Data Governance afford Maori the opportunity to be directly involved as kaitiaki of their mātauranga and assert rangatiratanga over data and its use.

year: 2021