The following research has focused on the utility of Ulva as an indicator of seawater nitrogen loading. Evaluation was made in three ways: 1) Observation of large-scale geographic variation in nitrogen status in natural populations around New Zealand in summer and winter, 2) Laboratory-based experimental assessment of the biochemical responses of N-indices in Ulva to nitrogen enrichment, and 3) Culturing standardized test-Ulva under low nutrient conditions which could be deployed into a variety of field situations. Seawater inorganic nutrient (nitrate, nitrite, ammonium and phosphate) concentrations and nitrogen (N)-indices (free amino acids, chlorophyll and total tissue nitrogen) in natural Ulva populations from 32 sites around New Zealand were compared. Sites were divided into 6 environmental categories: sheltered rural, exposed rural, rock pools, sheltered urban, exposed urban, and nitrogen-enriched urban sites. Seawater nutrient concentrations were highly variable between all sites in summer and winter. However, in the summer enriched urban sites had the highest mean total inorganic nitrogen concentrations and Ulva with the highest mean levels of all N-indices compared with any other environmental category. In the winter, Ulva contained more nitrogen (reflected in all N-indices) compared with Ulva in the summer, particularly in populations growing in colder southern seawater on more exposed coasts. The increase in Ulva N-status was not explained by increased seawater inorganic nitrogen concentrations. With univariate and multivariate statistical approaches it was shown that there was a significant effect of seawater temperature and site exposure on N-status in Ulva. Compared with other N-indices, stable nitrogen isotopes (δ15N) from Ulva growing in enriched urban sites had the widest range (4.77 ± 0.04 ‰ to 15.16 ± 0.03 ‰) of values compared with all other categories in both summer and winter. Conversely, Ulva from exposed rural sites had the lowest range of δ15N values compared with any other category (6.7 ± 0.1 to 8.8 ± 0.1 ‰) and showed no seasonal change in mean values (7.8 ‰ and 7.6 ‰ for summer and winter, respectively). In addition, δ15N values in Ulva were the only N-index that showed a significant difference between urban and rural categories. To test the relationship between inorganic nitrogen concentration in seawater and the responses of biochemical nitrogen indices in Ulva pertusa, several experiments were conducted in an outdoor, flow-through culture apparatus, in summer and winter. In this apparatus effects of ammonium concentration, nitrogen source (nitrate and ammonium), light and seawater motion were investigated. Of the same three N-indices examined in natural Ulva populations (free amino acids, chlorophyll and total tissue nitrogen), increases in free amino acids, particularly asparagine, provided the strongest indicator of increases in nitrogen availability. In addition, while tissue nitrogen and chlorophyll also increased with seawater nitrogen concentration, it was apparent that these indices were also strongly influenced by light, and probably season. Rates of ammonium assimilation provided no overall measure of the availability of nitrogen in seawater and were clearly affected by season. Similarly, growth rates in Ulva only showed a response to nitrogen addition in summer months. Stable isotopes of nitrogen (δ15N) in Ulva provided a clear distinction between natural and synthetic nitrogen sources, but more importantly, showed only minor fractionation (ranging from 1.3 ‰ to -1.9 ‰) of 15N supplied from synthetic nitrate and ammonium under both light-saturating and light-limiting conditions. To further develop Ulva as a standardized test-organism it was cultured in low-nutrient (non-polluted) seawater to deplete internal storage pools of nitrogen. Each month the resulting test-Ulva was then placed in surface-moored growth enclosures at a range of coastal sites around Auckland and then monitored for one year. In winter there were increases in seawater inorganic nitrogen concentrations and concomitant increases in free amino acid content. However, tissue nitrogen and chlorophyll content in test-Ulva showed similar increases (possibly saturating) across all sites suggesting that seasonal increases in these N-indices were also due to other seasonal factors (e.g., surface irradiance and / or seawater temperature). On the other hand, the total free amino acid pool showed strong differences between a low-nitrogen reference site and the other study sites all year round. It was probable that test-Ulva was integrating differences in tidally-averaged nitrogen loading that were not reliably detected in instantaneous seawater samples. In addition to N-indices in test-Ulva, levels of tissue heavy metals and stable isotopes of nitrogen showed strong differences with higher values of both typically found in urban environments compared with values found in non-polluted reference sites. It is concluded that several abiotic and biotic factors affect nitrogen status in Ulva, but the average nitrogen concentration in seawater, and the physical factors of temperature, light and water motion, appear to be the overarching determinants. It is further suggested that in combination with Ulva tissue δ15N values, tissue nitrogen and the free amino acid pool, as quantitative biochemical measures of nitrogen availability, are likely to provide useful information on both the amount and composition of nitrogen entering coastal environments. / Foundation for Research, Science and Technology. Auckland Regional Council.

The following research has focused on the utility of Ulva as an indicator of seawater nitrogen loading. Evaluation was made in three ways: 1) Observation of large-scale geographic variation in nitrogen status in natural populations around New Zealand in summer and winter, 2) Laboratory-based experimental assessment of the biochemical responses of N-indices in Ulva to nitrogen enrichment, and 3) Culturing standardized test-Ulva under low nutrient conditions which could be deployed into a variety of field situations. Seawater inorganic nutrient (nitrate, nitrite, ammonium and phosphate) concentrations and nitrogen (N)-indices (free amino acids, chlorophyll and total tissue nitrogen) in natural Ulva populations from 32 sites around New Zealand were compared. Sites were divided into 6 environmental categories: sheltered rural, exposed rural, rock pools, sheltered urban, exposed urban, and nitrogen-enriched urban sites. Seawater nutrient concentrations were highly variable between all sites in summer and winter. However, in the summer enriched urban sites had the highest mean total inorganic nitrogen concentrations and Ulva with the highest mean levels of all N-indices compared with any other environmental category. In the winter, Ulva contained more nitrogen (reflected in all N-indices) compared with Ulva in the summer, particularly in populations growing in colder southern seawater on more exposed coasts. The increase in Ulva N-status was not explained by increased seawater inorganic nitrogen concentrations. With univariate and multivariate statistical approaches it was shown that there was a significant effect of seawater temperature and site exposure on N-status in Ulva. Compared with other N-indices, stable nitrogen isotopes (δ15N) from Ulva growing in enriched urban sites had the widest range (4.77 ± 0.04 ‰ to 15.16 ± 0.03 ‰) of values compared with all other categories in both summer and winter. Conversely, Ulva from exposed rural sites had the lowest range of δ15N values compared with any other category (6.7 ± 0.1 to 8.8 ± 0.1 ‰) and showed no seasonal change in mean values (7.8 ‰ and 7.6 ‰ for summer and winter, respectively). In addition, δ15N values in Ulva were the only N-index that showed a significant difference between urban and rural categories. To test the relationship between inorganic nitrogen concentration in seawater and the responses of biochemical nitrogen indices in Ulva pertusa, several experiments were conducted in an outdoor, flow-through culture apparatus, in summer and winter. In this apparatus effects of ammonium concentration, nitrogen source (nitrate and ammonium), light and seawater motion were investigated. Of the same three N-indices examined in natural Ulva populations (free amino acids, chlorophyll and total tissue nitrogen), increases in free amino acids, particularly asparagine, provided the strongest indicator of increases in nitrogen availability. In addition, while tissue nitrogen and chlorophyll also increased with seawater nitrogen concentration, it was apparent that these indices were also strongly influenced by light, and probably season. Rates of ammonium assimilation provided no overall measure of the availability of nitrogen in seawater and were clearly affected by season. Similarly, growth rates in Ulva only showed a response to nitrogen addition in summer months. Stable isotopes of nitrogen (δ15N) in Ulva provided a clear distinction between natural and synthetic nitrogen sources, but more importantly, showed only minor fractionation (ranging from 1.3 ‰ to -1.9 ‰) of 15N supplied from synthetic nitrate and ammonium under both light-saturating and light-limiting conditions. To further develop Ulva as a standardized test-organism it was cultured in low-nutrient (non-polluted) seawater to deplete internal storage pools of nitrogen. Each month the resulting test-Ulva was then placed in surface-moored growth enclosures at a range of coastal sites around Auckland and then monitored for one year. In winter there were increases in seawater inorganic nitrogen concentrations and concomitant increases in free amino acid content. However, tissue nitrogen and chlorophyll content in test-Ulva showed similar increases (possibly saturating) across all sites suggesting that seasonal increases in these N-indices were also due to other seasonal factors (e.g., surface irradiance and / or seawater temperature). On the other hand, the total free amino acid pool showed strong differences between a low-nitrogen reference site and the other study sites all year round. It was probable that test-Ulva was integrating differences in tidally-averaged nitrogen loading that were not reliably detected in instantaneous seawater samples. In addition to N-indices in test-Ulva, levels of tissue heavy metals and stable isotopes of nitrogen showed strong differences with higher values of both typically found in urban environments compared with values found in non-polluted reference sites. It is concluded that several abiotic and biotic factors affect nitrogen status in Ulva, but the average nitrogen concentration in seawater, and the physical factors of temperature, light and water motion, appear to be the overarching determinants. It is further suggested that in combination with Ulva tissue δ15N values, tissue nitrogen and the free amino acid pool, as quantitative biochemical measures of nitrogen availability, are likely to provide useful information on both the amount and composition of nitrogen entering coastal environments. / Foundation for Research, Science and Technology. Auckland Regional Council.

The following research has focused on the utility of Ulva as an indicator of seawater nitrogen loading. Evaluation was made in three ways: 1) Observation of large-scale geographic variation in nitrogen status in natural populations around New Zealand in summer and winter, 2) Laboratory-based experimental assessment of the biochemical responses of N-indices in Ulva to nitrogen enrichment, and 3) Culturing standardized test-Ulva under low nutrient conditions which could be deployed into a variety of field situations. Seawater inorganic nutrient (nitrate, nitrite, ammonium and phosphate) concentrations and nitrogen (N)-indices (free amino acids, chlorophyll and total tissue nitrogen) in natural Ulva populations from 32 sites around New Zealand were compared. Sites were divided into 6 environmental categories: sheltered rural, exposed rural, rock pools, sheltered urban, exposed urban, and nitrogen-enriched urban sites. Seawater nutrient concentrations were highly variable between all sites in summer and winter. However, in the summer enriched urban sites had the highest mean total inorganic nitrogen concentrations and Ulva with the highest mean levels of all N-indices compared with any other environmental category. In the winter, Ulva contained more nitrogen (reflected in all N-indices) compared with Ulva in the summer, particularly in populations growing in colder southern seawater on more exposed coasts. The increase in Ulva N-status was not explained by increased seawater inorganic nitrogen concentrations. With univariate and multivariate statistical approaches it was shown that there was a significant effect of seawater temperature and site exposure on N-status in Ulva. Compared with other N-indices, stable nitrogen isotopes (δ15N) from Ulva growing in enriched urban sites had the widest range (4.77 ± 0.04 ‰ to 15.16 ± 0.03 ‰) of values compared with all other categories in both summer and winter. Conversely, Ulva from exposed rural sites had the lowest range of δ15N values compared with any other category (6.7 ± 0.1 to 8.8 ± 0.1 ‰) and showed no seasonal change in mean values (7.8 ‰ and 7.6 ‰ for summer and winter, respectively). In addition, δ15N values in Ulva were the only N-index that showed a significant difference between urban and rural categories. To test the relationship between inorganic nitrogen concentration in seawater and the responses of biochemical nitrogen indices in Ulva pertusa, several experiments were conducted in an outdoor, flow-through culture apparatus, in summer and winter. In this apparatus effects of ammonium concentration, nitrogen source (nitrate and ammonium), light and seawater motion were investigated. Of the same three N-indices examined in natural Ulva populations (free amino acids, chlorophyll and total tissue nitrogen), increases in free amino acids, particularly asparagine, provided the strongest indicator of increases in nitrogen availability. In addition, while tissue nitrogen and chlorophyll also increased with seawater nitrogen concentration, it was apparent that these indices were also strongly influenced by light, and probably season. Rates of ammonium assimilation provided no overall measure of the availability of nitrogen in seawater and were clearly affected by season. Similarly, growth rates in Ulva only showed a response to nitrogen addition in summer months. Stable isotopes of nitrogen (δ15N) in Ulva provided a clear distinction between natural and synthetic nitrogen sources, but more importantly, showed only minor fractionation (ranging from 1.3 ‰ to -1.9 ‰) of 15N supplied from synthetic nitrate and ammonium under both light-saturating and light-limiting conditions. To further develop Ulva as a standardized test-organism it was cultured in low-nutrient (non-polluted) seawater to deplete internal storage pools of nitrogen. Each month the resulting test-Ulva was then placed in surface-moored growth enclosures at a range of coastal sites around Auckland and then monitored for one year. In winter there were increases in seawater inorganic nitrogen concentrations and concomitant increases in free amino acid content. However, tissue nitrogen and chlorophyll content in test-Ulva showed similar increases (possibly saturating) across all sites suggesting that seasonal increases in these N-indices were also due to other seasonal factors (e.g., surface irradiance and / or seawater temperature). On the other hand, the total free amino acid pool showed strong differences between a low-nitrogen reference site and the other study sites all year round. It was probable that test-Ulva was integrating differences in tidally-averaged nitrogen loading that were not reliably detected in instantaneous seawater samples. In addition to N-indices in test-Ulva, levels of tissue heavy metals and stable isotopes of nitrogen showed strong differences with higher values of both typically found in urban environments compared with values found in non-polluted reference sites. It is concluded that several abiotic and biotic factors affect nitrogen status in Ulva, but the average nitrogen concentration in seawater, and the physical factors of temperature, light and water motion, appear to be the overarching determinants. It is further suggested that in combination with Ulva tissue δ15N values, tissue nitrogen and the free amino acid pool, as quantitative biochemical measures of nitrogen availability, are likely to provide useful information on both the amount and composition of nitrogen entering coastal environments. / Foundation for Research, Science and Technology. Auckland Regional Council.