title: Development of microsatellite markers for Antarctic Bryum Hedw. species

authors: Harfoot, Rhodri T.

abstract: The objective of this thesis was to isolate microsatellite markers using the Glenn (2001) method from the species Bryum argenteum so as to be able to study these markers in Antarctic populations of Bryum species. Microsatellite regions have been found to be highly polymorphic and neutral markers, and usually genus specific, thus making them ideal for population genetic studies. The populations to be studied in the future have a large distribution over the Southern Victoria Land area, ranging from the Dry Valleys to Granite Harbour and Ross Island. 
Mosses are the most abundant and widespread of the vascular plant groups within continental Antarctica. They inhabit locations that are some of the more extreme on earth and experience periods of desiccation and darkness that can last as long as four months. For these reasons the establishment of mosses in Antarctica is a subject that has attracted great debate. One hypothesis suggests that mosses first became established when the ice retreated from the land approximately 17000 years ago, at the end of the last glacial maximum. The alternative hypothesis is that mosses survived as relictual populations over this period and have recently increased their habitat range. The study of microsatellite length polymorphism in populations will allow these hypotheses to be tested. 
The genus Bryum Hedw. (Bryaceae) is a highly polymorphic, cosmopolitan genus that is abundant in Antarctica over a wide range of locations. It is found from the Sub-Antarctic zone (Sub-Antarctic islands) to the continental zone (Continental Antarctica and Southern and Eastern Antarctic Peninsula. Thus covering a wide range of habitats from warm and wet (e.g. the Sub-Antarctic islands) to cold and very dry (e.g. the McMurdo Dry Valleys). 
To study the population genetics of Antarctic Bryum species, development of microsatellite markers was necessary as it has been found that with less specific methods such as RAPD-PCR, the DNA used for the analyses had been contaminated by co-extracted DNA from fungi living on the mosses, thus confounding the results obtained. Microsatellites, once developed, are genus or family specific, thus there is little risk of amplifying a contaminant when using microsatellite markers. Abstract 
This project failed to isolate any microsatellite markers from Bryum argenteum, due to experimental difficulties that occurred at three major stages; ligation, transformation and hybridisation screening of the genomic library. Future research should be focussed on completion of microsatellite isolation for this genus and on evaluation of the population relationships among Antarctic localities.

text: Development of microsatellite markers for Antarctic Bryum Hedw. species The objective of this thesis was to isolate microsatellite markers using the Glenn (2001) method from the species Bryum argenteum so as to be able to study these markers in Antarctic populations of Bryum species. Microsatellite regions have been found to be highly polymorphic and neutral markers, and usually genus specific, thus making them ideal for population genetic studies. The populations to be studied in the future have a large distribution over the Southern Victoria Land area, ranging from the Dry Valleys to Granite Harbour and Ross Island. 
Mosses are the most abundant and widespread of the vascular plant groups within continental Antarctica. They inhabit locations that are some of the more extreme on earth and experience periods of desiccation and darkness that can last as long as four months. For these reasons the establishment of mosses in Antarctica is a subject that has attracted great debate. One hypothesis suggests that mosses first became established when the ice retreated from the land approximately 17000 years ago, at the end of the last glacial maximum. The alternative hypothesis is that mosses survived as relictual populations over this period and have recently increased their habitat range. The study of microsatellite length polymorphism in populations will allow these hypotheses to be tested. 
The genus Bryum Hedw. (Bryaceae) is a highly polymorphic, cosmopolitan genus that is abundant in Antarctica over a wide range of locations. It is found from the Sub-Antarctic zone (Sub-Antarctic islands) to the continental zone (Continental Antarctica and Southern and Eastern Antarctic Peninsula. Thus covering a wide range of habitats from warm and wet (e.g. the Sub-Antarctic islands) to cold and very dry (e.g. the McMurdo Dry Valleys). 
To study the population genetics of Antarctic Bryum species, development of microsatellite markers was necessary as it has been found that with less specific methods such as RAPD-PCR, the DNA used for the analyses had been contaminated by co-extracted DNA from fungi living on the mosses, thus confounding the results obtained. Microsatellites, once developed, are genus or family specific, thus there is little risk of amplifying a contaminant when using microsatellite markers. Abstract 
This project failed to isolate any microsatellite markers from Bryum argenteum, due to experimental difficulties that occurred at three major stages; ligation, transformation and hybridisation screening of the genomic library. Future research should be focussed on completion of microsatellite isolation for this genus and on evaluation of the population relationships among Antarctic localities.

year: 2002

title: Dairy Effluent Steroidal Hormones Characteristics and Treatment by Anaerobic Digestion (Covered Anaerobic Pond) and Biochar

authors: Rachmani, Asaf

subjects: Dairy effluent treatment system

abstract: Endocrine disrupting compounds such as estrogenic steroid hormones (ESHs) are problematic when present in waterways in terms of both the health of the ecosystem and downstream water treatment. Dairy farms are some of the largest contributors to ESHs in the environment. This study focused on a farm located in the Waikato region, New Zealand, which housed 550, primarily grass-fed dairy cows. The farm included a rotary milking shed, feed pad, sump, covered anaerobic pond and storage pond, with effluent and sludge being applied to land. In this study concentrations of the conjugated and unconjugated ESHs, 17α-estradiol (17α-E2), 17β-estradiol (17β-E2) and estrone (E1), in both dissolved and solid phases of dairy shed effluent, and covered anaerobic pond sludge and effluent, were measured from grab samples over a period of nine months. These were used to investigate seasonal variation and performance of the covered anaerobic pond in removing the ESHs and their conversion from one form into another. Previously published methods of ESH analysis were validated and adapted for ESH analysis in dissolved and solid phases, and a new enzymatic method for conjugate ESH analysis was tested and used. In addition, the effect of dosing the effluent with biochar was examined on ESH removal. A covered anaerobic pond model was developed to examine the efficacy of a dairy farm-based covered anaerobic pond treatment system to settle, transform, absorb and remove ESHs. The model was calibrated utilising ESH measurements and used to examine the effect of biochar addition and operational parameters on ESH removal. 

Overall ESH concentrations in the covered anaerobic pond influent, sludge and effluent were 4,171 ng/L, 93,601 ng/L and 4,346 ng/L respectively. ESH concentrations peaked in dairy shed effluent during April and July, which correlated with the late pregnancy and calving periods on the farm. The mean organic carbon normalised adsorption coefficient (Koc) for ESHs in the samples from the pond treatment system ranged between 3.06 mL/g to 3.78 mL/g, comparable with published values in soil and wastewater sludge. Up to 99% of the total mass of ESHs in the sludge and between 70-80 % in influent and effluent were retained in the solids phase. The predominant ESH in the influent samples was 17α-E2 (2,869 ng/L), but E1 predominated in the sludge (85,414 ng/L) and in the effluent (3,140 ng/L). The dissolved and solid phases of dairy shed effluent contained the highest relative proportions of conjugated ESHs with means of 25.1 % and 3.38 % respectively and corresponding mean concentrations of 113 ng/L and 137 ng/L. In contrast, sludge and effluent samples (dissolved and solid phases) from the covered anaerobic pond contained smaller relative proportions of conjugated ESHs, 0.90 % and 1.21 %, and 7.17 % and 0.43 % respectively and corresponding mean concentrations of 6.65 ng/L and 967 ng/L, and 52.5 ng/L and 19.7 ng/L respectively. These results demonstrate the importance of considering the solid fraction within effluent treatment systems when analysing ESHs, otherwise estrogenic load can be greatly underestimated. The conjugated ESHs were present in both the dissolved and solid phases of all samples collected, indicating that conjugated ESHs are persistent and can pass through anaerobic treatment systems contributing to estrogenic load once applied to pasture, and potentially leach into groundwater or migrate to nearby surface waters. Overall anaerobic treatment of dairy waste decreased the contribution of 17α-E2 and 17-E2 while increasing that of E1, however, 17-E2 was the main contributor to total estrogenicity.

The calibration of the model developed provided a good fit with experimental data with an R2 of 0.98-0.99. Addition of biochar into dairy shed effluent and the model covered anaerobic pond system resulted in an 89 % reduction of free ESHs but had a minimal impact on conjugated ESH. Addition of biochar increased the solid phase ESH concentration by 19 % settling out into the sludge and reducing overall estrogenic load in the effluent. Operational factors such as higher influent flow rates and sludge accumulation negatively impacted the covered anaerobic pond ESH removal performance. Sludge accumulation and short-circuiting caused by infrequent removal and shallow depth of the covered anaerobic pond system resulted in mixing of fresh influent with the upper layer of pond sludge, leading to decreased removal efficiency. To enhance the covered anaerobic pond system's performance, increasing the retention time and reducing sludge carry over by increasing pond volume and depth, resulted in ESH removal increasing to 67.9 % (no biochar) and 73.0 % (with biochar), and estrogenicity reduction improving to 70.4 % (no biochar) and 73.5 % (with biochar).

text: Dairy Effluent Steroidal Hormones Characteristics and Treatment by Anaerobic Digestion (Covered Anaerobic Pond) and Biochar Endocrine disrupting compounds such as estrogenic steroid hormones (ESHs) are problematic when present in waterways in terms of both the health of the ecosystem and downstream water treatment. Dairy farms are some of the largest contributors to ESHs in the environment. This study focused on a farm located in the Waikato region, New Zealand, which housed 550, primarily grass-fed dairy cows. The farm included a rotary milking shed, feed pad, sump, covered anaerobic pond and storage pond, with effluent and sludge being applied to land. In this study concentrations of the conjugated and unconjugated ESHs, 17α-estradiol (17α-E2), 17β-estradiol (17β-E2) and estrone (E1), in both dissolved and solid phases of dairy shed effluent, and covered anaerobic pond sludge and effluent, were measured from grab samples over a period of nine months. These were used to investigate seasonal variation and performance of the covered anaerobic pond in removing the ESHs and their conversion from one form into another. Previously published methods of ESH analysis were validated and adapted for ESH analysis in dissolved and solid phases, and a new enzymatic method for conjugate ESH analysis was tested and used. In addition, the effect of dosing the effluent with biochar was examined on ESH removal. A covered anaerobic pond model was developed to examine the efficacy of a dairy farm-based covered anaerobic pond treatment system to settle, transform, absorb and remove ESHs. The model was calibrated utilising ESH measurements and used to examine the effect of biochar addition and operational parameters on ESH removal. 

Overall ESH concentrations in the covered anaerobic pond influent, sludge and effluent were 4,171 ng/L, 93,601 ng/L and 4,346 ng/L respectively. ESH concentrations peaked in dairy shed effluent during April and July, which correlated with the late pregnancy and calving periods on the farm. The mean organic carbon normalised adsorption coefficient (Koc) for ESHs in the samples from the pond treatment system ranged between 3.06 mL/g to 3.78 mL/g, comparable with published values in soil and wastewater sludge. Up to 99% of the total mass of ESHs in the sludge and between 70-80 % in influent and effluent were retained in the solids phase. The predominant ESH in the influent samples was 17α-E2 (2,869 ng/L), but E1 predominated in the sludge (85,414 ng/L) and in the effluent (3,140 ng/L). The dissolved and solid phases of dairy shed effluent contained the highest relative proportions of conjugated ESHs with means of 25.1 % and 3.38 % respectively and corresponding mean concentrations of 113 ng/L and 137 ng/L. In contrast, sludge and effluent samples (dissolved and solid phases) from the covered anaerobic pond contained smaller relative proportions of conjugated ESHs, 0.90 % and 1.21 %, and 7.17 % and 0.43 % respectively and corresponding mean concentrations of 6.65 ng/L and 967 ng/L, and 52.5 ng/L and 19.7 ng/L respectively. These results demonstrate the importance of considering the solid fraction within effluent treatment systems when analysing ESHs, otherwise estrogenic load can be greatly underestimated. The conjugated ESHs were present in both the dissolved and solid phases of all samples collected, indicating that conjugated ESHs are persistent and can pass through anaerobic treatment systems contributing to estrogenic load once applied to pasture, and potentially leach into groundwater or migrate to nearby surface waters. Overall anaerobic treatment of dairy waste decreased the contribution of 17α-E2 and 17-E2 while increasing that of E1, however, 17-E2 was the main contributor to total estrogenicity.

The calibration of the model developed provided a good fit with experimental data with an R2 of 0.98-0.99. Addition of biochar into dairy shed effluent and the model covered anaerobic pond system resulted in an 89 % reduction of free ESHs but had a minimal impact on conjugated ESH. Addition of biochar increased the solid phase ESH concentration by 19 % settling out into the sludge and reducing overall estrogenic load in the effluent. Operational factors such as higher influent flow rates and sludge accumulation negatively impacted the covered anaerobic pond ESH removal performance. Sludge accumulation and short-circuiting caused by infrequent removal and shallow depth of the covered anaerobic pond system resulted in mixing of fresh influent with the upper layer of pond sludge, leading to decreased removal efficiency. To enhance the covered anaerobic pond system's performance, increasing the retention time and reducing sludge carry over by increasing pond volume and depth, resulted in ESH removal increasing to 67.9 % (no biochar) and 73.0 % (with biochar), and estrogenicity reduction improving to 70.4 % (no biochar) and 73.5 % (with biochar).

year: 2024

title: Bilingual/Immersion education: Indicators of good practice

authors: May, Stephen

subjects: Maori

abstract: Based on a review of national and international research on bilingualism and bilingual/immersion education, this report explores effective approaches for bilingual education. While the focus was on Māori-medium education, the indicators of good practice can also be applied to other bilingual contexts in Aotearoa/New Zealand, such as Pasifika bilingual education.

text: Bilingual/Immersion education: Indicators of good practice Based on a review of national and international research on bilingualism and bilingual/immersion education, this report explores effective approaches for bilingual education. While the focus was on Māori-medium education, the indicators of good practice can also be applied to other bilingual contexts in Aotearoa/New Zealand, such as Pasifika bilingual education.

year: 2004