A series of experiments were conducted to determine nutritional strategies to improve the efficiency of N utilization in dairy cows when feeding co-products including wheat-based (W-DDGS) and corn-wheat blend distillers grains with solubles (B-DDGS), and dried whey permeate (DWP). In Experiment 1, the objective was to determine the effects of replacing canola meal (CM) as the major protein source with W-DDGS on ruminal fermentation, microbial protein production, omasal nutrient flow, and animal performance. Cows were fed either a standard barley silage-based total mixed ration containing CM as the major protein supplement (0% W-DDGS, control) or diets formulated to contain 10, 15 and 20% W-DDGS (dry matter [DM] basis), with W-DDGS replacing primarily CM. Diets were isonitrogenous (18.9% crude protein [CP]). Inclusion of W-DDGS to the diet did not negatively affect ruminal fermentation, microbial protein production, and omasal nutrient flow. However, there was a 0.7- to 2.4-kg increase in DM intake, and a 1.2- to 1.8-kg increase in milk yield after the addition of W-DDGS in place of CM. In Experiment 2, the objective was to delineate the effects of including either W-DDGS or B-DDGS dried distillers grains with solubles as the major protein source in low or high CP diets fed to dairy cows on ruminal function, microbial protein synthesis, omasal nutrient flows, urea-N recycling, and milk production. The treatment factors were type of distillers co-product (W-DDGS vs. B-DDGS) and dietary CP content (15.2 vs. 17.3%; DM basis). The B-DDGS was produced from a mixture of 15% wheat and 85% corn grain. All diets were formulated to contain 10% W-DDGS or B-DDGS on a DM basis. Feeding up to 10% of dietary DM as B-DDGS or W-DDGS as the major source of protein did not have negative effects on metabolizable protein (MP) supply and milk production in dairy cows. However, reducing dietary CP content from 17.3 to 15.2% decreased milk production. This response was attributed to an insufficient supply of ruminally degradable protein (RDP) that suppressed microbial nonammonia N (NAN) synthesis in the rumen, thus decreasing intestinal MP supply. In Experiment 3, the objective was to determine the effects of replacing barley or corn starch with lactose (as DWP) in diets containing 10% W-DDGS on ruminal function, omasal nutrient flow, and lactation performance. The treatment factors were source of starch (barley vs. corn) and dietary inclusion level of DWP (0 vs. 6%; DM basis) as a partial replacement for starch. Diets were isonitrogenous (18% CP) and contained 3 or 8% total sugar. The starch content of the low sugar diet was 24% compared to 20% for the high sugar diet. Dry matter intake, and milk and milk component yields did not differ with diet. However, partially replacing dietary corn or barley starch with sugar up-regulated ruminal acetate and propionate absorption, and reduced ruminal NH3-N concentration, but had no effect on ruminal pH, microbial protein synthesis, omasal nutrient flow and production in dairy cows. In summary, data presented in this thesis indicate that W-DDGS and B-DDGS can be included as the major source of protein in dairy cow diets without compromising ruminal function, nutrient supply and milk production in dairy cows. Feeding medium to low CP diets, and partial replacement of starch with sugar in diets containing W-DDGS and B-DDGS can improve N utilization efficiency in dairy cows. Additionally, an upregulation of facilitated transport of acetate and propionate across epithelial cells possibly prevents the occurrence of ruminal acidosis when lactose partially replaces starch in cow diets.

A series of experiments were conducted to investigate how dietary and ruminal factors regulate urea-N recycling in ruminants. In Experiments 1, 2, and 3, urea-N kinetics were measured using 4-d intra-jugular infusions of [15N15N]-urea. In Experiment 1, the objective was to determine how interactions between dietary ruminally-degradable protein (RDP) level and ruminally-fermentable carbohydrate (RFC) may alter urea-N transfer to the gastrointestinal tract (GIT) and the utilization of this recycled urea-N in rapidly-growing lambs fed high N diets. The dietary factors were: 1) dry-rolled barley (DRB) vs. pelleted barley (PB) as the principal source of RFC; and 2) dietary levels of RDP of 60 vs. 70% (% of CP). Nitrogen intake, fecal and urinary N excretion increased as dietary RDP level increased; however, method of barley processing had no effect on N use. Dietary treatment had no effect on urea-N kinetics; however, endogenous production of urea-N (UER) exceeded N intake. For all diets, 0.669 to 0.742 of UER was recycled to the GIT; however, 0.636 to 0.756 of the GER was returned to the ornithine cycle. In Experiment 2, the objective was to delineate the effects of partial defaunation of the rumen on urea-N kinetics in lambs fed low or high N diets. Treatments were: 1) partial defaunation (PDFAUN) vs. faunation (FAUN); and 2) low (10%, LOW) vs. high (15%, HIGH) dietary CP. Linoleic acid-rich sunflower oil was fed as a partially-defaunating agent. Partial defaunation decreased ruminal NH3-N concentrations. The UER and urinary urea-N excretion (UUE) were lower, and the GER tended to be lower in PDFAUN as compared to FAUN lambs; however, as a proportion of UER, GER was higher and the proportion of recycled urea-N that was utilized for anabolism (i.e., UUA) tended to be higher in PDFAUN lambs. The UER, GER and UUE were higher in lambs fed diet HIGH; however, as a proportion of UER, GER and its anabolic use were higher in lambs fed diet LOW. In Experiment 3, the objective was to delineate how, at similar N intakes, interactions between ruminal partial defaunation and altering dietary RFC may alter urea-N kinetics and N metabolism in lambs. Treatments were: 1) PDFAUN vs. FAUN; and 2) DRB vs. PB. Urinary N excretion was lower and retained N was higher in PDFAUN compared to FAUN lambs. The UER was similar across treatments; however, the GER, expressed as absolute amounts or as a proportion of UER, UUA, and microbial N supply were higher in PDFAUN compared to FAUN lambs. As a proportion of UER, GER was higher, whereas UUE was lower in lambs fed PB compared to those fed DRB. In Experiment 4, the objective was to determine the effects of feeding oscillating dietary CP compared to static dietary CP concentration on N retention and in vitro urea flux across ruminal epithelia. Dietary treatments consisted of a medium CP diet (MEDIUM; 12.8% CP) or diets with oscillating CP content (OSC) fed in two different sequences i.e., 2 d of low CP (9.7% CP) followed by 2 d of high CP (16.1% CP; OSC-HIGH) or vice-versa (OSC-LOW). Ruminal epithelial tissues were collected and mounted in Ussing chambers under short-circuit conditions and the serosal-to-mucosal urea flux (Jsm-urea) was measured using 14C-urea. Although N intake was similar, retained N and microbial N supply were greater in lambs fed the OSC diets compared to those fed the MEDIUM diet. The total Jsm-urea was higher in lambs fed the OSC-LOW compared to those fed the OSC-HIGH diet. Across diets, the addition of phloretin (a known specific inhibitor of facilitative urea transporter-B; UT-B) reduced Jsm-urea; however, phloretin-insensitive Jsm-urea was the predominant route for transepithelial urea transfer. In summary, data presented in this thesis provide new insights that the improved N retention typically observed in defaunated ruminants and in ruminants fed oscillating dietary CP concentrations is partly mediated via increased urea-N recycling to the GIT and utilization of recycled urea-N for anabolic purposes.

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.

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.

FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / A pesquisa foi conduzida no Setor de Suinocultura do Centro de CiÃncias AgrÃrias do Departamento de Zootecnia da Universidade Federal do CearÃ. O experimento teve a duraÃÃo de 42 dias e foi realizado com o objetivo de avaliar o desempenho zootÃcnico, a histomorfometria intestinal, o perfil microbiolÃgico fecal, a ocorrÃncia de diarrÃia, os indicadores sÃricos do metabolismo do nitrogÃnio e o custo de produÃÃo de leitÃes submetidos a dietas contendo diferentes fontes de proteÃna de origem animal e vegetal durante a fase 1 (21-42 dias de idade) e 2 (42-63 dias de idade) do perÃodo de creche. Participaram do ensaio, inicialmente, 80 leitÃes machos linhagem comercial desmamados aos 21 dias de idade e com peso mÃdio de 5,49kg. O delineamento experimental utilizado foi em blocos ao acaso, com 5 tratamentos, 4 repetiÃÃes e 4 animais por repetiÃÃo na 1 semana, 3 animais por repetiÃÃo na 2 semana e 2 animais por repetiÃÃo na 3 semana (final da fase 1) e por toda a fase 2, sendo feito o desdobramento num fatorial 2 x 2 + 1, composto por duas fontes protÃicas animal (farinha de carne - FC e plasma sanguÃneo em pà - PSP), duas fontes protÃicas vegetal (farelo de algodÃo - FA e farelo da amÃndoa da castanha de caju - FACC) e um adicional (dieta controle - T1). Quando a anÃlise de variÃncia detectou significÃncia entre os tratamentos, foram aplicados contrastes para testar os efeitos dos fatores. AlÃm disso, a comparaÃÃo de mÃdias para proteÃna de origem animal (POA) e proteÃna de origem vegetal (POV) com a dieta controle foi feita atravÃs do teste de Dunnet. As dietas experimentais foram isonutritivas com 22% de proteÃna bruta (PB) e 3.500 kcal ED/kg na fase 1, sendo T1 â Dieta controle (DC) + 10% de leite desnatado em pà (LDP); T2 â DC + 5% PSP+ 15% FA; T3 - DC + 5% FC + 15% FA; T4 â DC + 5% PSP + 15% FACC e T5 - DC + 5% FC + 15% FACC. Para a fase 2 os tratamentos continham 21% PB e 3.400 kcal ED/kg, sendo T1 â Dieta controle (DC) + 5% LDP; T2 â DC + 4% PSP+ 12% FA; T3 - DC + 4% FC + 12% FA; T4 â DC + 4% PSP + 12% FACC e T5 - DC + 4% FC + 12% FACC. Foi concluÃdo que a substituiÃÃo do LDP pelo PSP ou pela FC, como fontes protÃicas de origem animal, à viÃvel com respeito aos parÃmetros histomorfomÃtricos, na primeira e segunda semana da fase 1; contagem das colÃnias fecais (CCF) e concentraÃÃo plasmÃtica das proteÃnas totais (CPPT), ao final das fases 1 e 2; e concentraÃÃo plasmÃtica de urÃia (CPU), ao final da fase 2. Entretanto, para os parÃmetros de desempenho zootÃcnico, em ambas as fases; e ocorrÃncia de diarrÃia, na primeira semana da fase 1, apenas o PSP mostrou-se viÃvel. A inclusÃo de 15% de FA ou 15% de FACC, em substituiÃÃo parcial ao farelo de soja, como fonte protÃica de origem vegetal, à satisfatÃria em relaÃÃo aos parÃmetros histomorfomÃtricos, na primeira e segunda semana da fase 1; CCF, CPPT e CPU, ao final das fases 1 e 2; alÃm da ocorrÃncia de diarrÃia, na primeira semana da fase 1. Contudo para os parÃmetros de desempenho zootÃcnico apenas o FA revelou-se viÃvel em ambas as fases. NÃo foram registradas interaÃÃes significativas (P>0,05) entre os fatores (proteÃna de origem animal â POA x proteÃna de origem vegetal - POV) para os parÃmetros de desempenho zootÃcnico, nas fases 1 e 2; para a CCF e CPPT, ao final da fase 1 e 2; alÃm da CPU, ao final da fase 2. Entre os fatores (semana x dieta experimental) nÃo foram verificadas interaÃÃes significativas (P>0,05) para os parÃmetros histomorfomÃtricos. Todavia para a CPU, ao final da fase 1, foram constatadas interaÃÃes significativas (P<0,05). Na fase 1, a dieta controle, e as dietas contendo FC, como POA, e FACC, como POV, proporcionaram o maior nÃmero de cepas microbianas nas fezes dos leitÃes. Na fase 2, as dietas contendo PSP, como POA, e FA, como POV, proporcionaram o maior nÃmero de cepas microbianas nas fezes dos leitÃes. A melhor resposta econÃmica para produÃÃo de leitÃes no perÃodo de creche foi obtida com a dieta contendo PSP e FA (T2) / The research was developed in the Sector of Swine of the Department of Zootechnia of the Center of Agrarian Sciences of the Federal University of CearÃ. The experiment had the duration of 42 days and it was accomplished with the objective of evaluating the zootechnic performance, the intestinal histomorfometry, the fecal microbiological profile, the diarrhea occurrence, the serical indicators of the nitrogen metabolism and the production cost of piglets submitted to diets containing different sources of animal and vegetable protein origin during the phases 1 (21-42 days of age)and 2 (42-63 days of age) in the nursery period. They participated in the assay, initially, 80 male piglets of commercial lineage weaned at 21 days of age and with medium weight of 5,49kg. The experiment followed randomized blocks, with 5 treatments, 4 repetitions and 4 animals per repetition in the 1st week, 3 animals per repetition in the 2nd week and 2 animals per repetition in the 3rd week (end of phase 1) and for the whole phase 2, being made the unfolding in a factorial 2 x 2 + 1, composed by two sources of animal protein (meat meal - MM and powdered sanguine plasma - PSP), two sources of vegetal protein (cotton meal - CM and cashew nut meal - CNM) and an additional one (diet control - T1). When the variance analysis detected significance among the treatments, contrasts were applied to test the effects of the factors. Besides, the comparison of averages for protein of animal origin (PAO) and protein of vegetal origin (PVO) with the diet control which was done through the Dunnet tests. The diets were isonutritives with crude protein (CP) of 22% and level of energy of 3.500 kcal DE/kg in the phase 1, being T1 - Diet control (DC) + 10% of skimmed powdered milk (SPM); T2 - DC + 5% PSP+ 15% CM; T3 - DC + 5% MM + 15% CM; T4 - DC + 5% PSP + 15% CNM and T5 - DC + 5% MM + 15% CNM. For the phase 2 the treatments contained 21% CP and 3.400 kcal DE/kg, being T1 - Diet control (DC) + 5% SPM; T2 - DC + 4% PSP+ 12% CM; T3 - DC + 4% MM + 12% CM; T4 - DC + 4% PSP + 12% CNM and T5 - DC + 4% MM + 12% CNM. It was concluded that the substitution of SPM for PSP or for MM, as sources of protein of animal origin, is viable with regard to the histomorfometric parameters in the first and second week of phase 1; counting of the fecal colonies (CFC) and plasmatic concentration of total proteins (PCTP), at the end of phases 1 and 2; and plasmatic concentration of urea (PCU), at the end of phase 2. Nevertheless, for the parameters of zootechnic performance, in both phases; and diarrhea occurrence, in the first week of phase 1, just PSP was shown viable. The inclusion of 15% of CM or 15% of CNM, in partial substitution by the soybean meal, as source of vegetal origin protein, is satisfactory in relation to the histomorfometric parameters in the first and second week of phase 1; CFC, PCTP and PCU, at the end of phases 1 and 2; besides the diarrhea occurrence, in the first week of phase 1. However for the zootechnic parameters only the CM was revealed viable in both phases. Significant interactions were not registered (P>0,05) among the factors (protein of animal origin - PAO x protein of vegetal origin - PVO) for the zootechnic performance parameters, in phases 1 and 2, to the CFC and PCTP, at the end of phases 1 e 2; besides PCU, at the end of the phase 2. Among the factors (week x experimental diet) significant interactions were not verified (P>0,05) for the histomorfometric parameters. Though for PCU, at the end of the phase 1, significant interactions were verified (P <0,05). In the phase 1, the diet control, and the diets containing MM, like PAO, and CNM, like PVO provided the largest number of microbial stumps in the feces of the piglets. In phase 2, the diets containing PSP, as PAO, and CM, like PVO the largest number of microbial stumps in the feces of the piglets was provided. The best economical answer for production of piglets in the nursery period was obtained with the diet containing PSP and CM (T2)

FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / O ensaio foi realizado no Setor de Suinocultura do Centro de CiÃncias AgrÃrias do Departamento de Zootecnia da Universidade Federal do CearÃ, com o objetivo de avaliar o desempenho zootÃcnico, os indicadores do metabolismo do nitrogÃnio, o bem-estar animal e o custo de produÃÃo de leitÃes no prÃodo de creche, submetidos a dietas à base de sorgo-soja enriquecidas com caseÃna + lactose isoladas. O experimento teve a duraÃÃo de 42 dias, sendo dividido em duas fases experimentais, a saber: fase 1(21-42 dias) e fase 2 (42 a 63 dias). Foram utilizados 40 animais de linhagem comercial, desmamados com idade aproximada de 21 dias, apresentando um peso mÃdio de 4,76kg. O delineamento experimental utilizado foi em blocos ao acaso, com 4 tratamentos e 5 repetiÃÃes, apresentando 2 animais por repetiÃÃo. Foi realizado um desdobramento fatorial 2X2, sendo 2 cereais energÃticos (milho e sorgo) e 2 suplementos lÃcteos (leite desnatado em pà e caseÃna + lactose). A comparaÃÃo entre as mÃdias foi realizada pelo teste de Duncan a 5%. As dietas experimentais foram isoenergÃticas, isoprotÃicas e isonutrientes para lisina e met+cis. Na fase 1 foi utilizado 21 % de PB e 3350 kcal de EM &#8260; kg. e na fase 2, 18,5 % de PB e 3250kcal de EM &#8260; kg. As dietas experimentais foram em nÃmero de 4 por fase, da seguinte forma: T1 (milho +LDP), T2 (milho+caseÃna +lactose), T3 (sorgo+LDP) e T4 (sorgo+caseÃna+lactose). Foi concluÃdo que a substituiÃÃo do milho pelo sorgo granÃfero e a inclusÃo da lactose + caseÃna à viÃvel quanto ao desempenho zootÃcnico. A utilizaÃÃo do sorgo e da caseÃna+lactose nas dietas para leitÃes desmamados nÃo ocasionou diferenÃas significativas para os indicadores do metabolismo do nitrogÃnio nas duas fases experimentais. Os tratamentos nÃo influenciaram os indicadores do bem-estar animal. NÃo foram registradas interaÃÃes significativas (P>0,05) entre os alimentos energÃticos e os alimentos lÃcteos, nas fases 1 e 2 do perÃodo de creche, para o desempenho zootÃcnico e para os indicadores do bem-estar animal. A melhor resposta econÃmica para a produÃÃo de leitÃes no perÃodo de creche foi obtida com a dieta contendo sorgo + caseÃna +lactose (T4). / The research was developed in the Division of Swine Pro duction, Department of Animal Science, Agricultural Science Center, Federal Un iversity of CearÃ, with the objective of evaluating the animal performance, the occurrence diarrhea, the nitrogen metabolism indicators, animal well-being, and the production cost of piglets submitted to diets containing sorghum-soybean me al and isolated casein+lactose. The assay had the duration of 42 days and it was divided in two phases: 1 (21-42 days of age) and 2 (42-63 days of age ) during the nursery period. The experiment used a total of 40 piglets of a commercial line, weaned at 21 days of age (average weight of 4,76 kg). The exper iment followed a randomized blocks design, with 4 treatments, 5 repetitio ns and 2 animals per repetition, unfolded in a factorial 2 x 2 design, comp osed by two energetic feeds (corn and sorghum) and two milk products (dried skim milk a nd casein + lactose). Mean values were compared through the Duncan's tests 5 %. The diets were isoenergetics, isoproteics and isonutritives for lysine and methionine + cystine. In phase 1, a level of 21% crude protein (CP) and 3350 kcal ME was used and in phase 2, a level of 18,5 % crude protein (CP) and 32 50 kcal ME was used. The experimental diets were in number of 4 for phase: T 1 (corn+ dried skim milk), T 2 (corn + casein + lactose), T 3 (sorghum + dried skim milk) and T 4 (sorghum + casein + lactose). The use of sorghum and casein + lactose in weaned pig diets did not cause significant differences for nitrogen met abolism indicators, in both phases of assay. The treatments did not influence the w ell-being indicators. Related to animal performance and well-being no signi ficant interactions were registered (P>0,05) among the factors, during phases 1 a nd 2 of the nursery xxi period. The best economical answer for production of pi glets in the nursery period was obtained with the diet containing sorghum + casein + lactose (T4).

Uncontrolled spreading of genetically modified (GM) plants is one of the main concerns about their cultivation. Inducible RNA interference against an essential gene could be a tool for control of GM plants. After spraying with a chemical inducer, the essential gene will be silenced so the treated GM plant will die. For testing this strategy we chose two key enzymes of nitrogen metabolism, glutamate synthase (GOGAT) and glutamine synthetase (GS). GS processes ammonium ions into glutamine, then GOGAT transfers the amide group from glutamine to 2-oxoglutarate to form two glutamates. GS/GOGAT cycle is the main pathway for assimilation of ammonium ions, which could be toxic to plants in a higher concentration. Disruption of ammonium assimilation during photorespiration causes a strong inhibition of photosynthesis. The aim of this work was to describe the effects of silencing GOGAT and GS genes in Arabidopsis thaliana. To induce silencing, RNAi hairpin constructs under a control of constitutive or estradiol-inducible promoter were prepared. In selected independent transformants with the inducible hairpin against GOGAT, chlorosis and reduced growth were observed after the estradiol treatment in in vitro conditions. However, the spraying with estradiol was tricky, at the whole plant level, the induction of...

No description available.