Arginine ammonification assay as a rapid index of gross N mineralization in agricultural soils

Seasonal dynamics of in situ gross nitrogen (N) mineralization rates were measured using the ¹⁵N-NH₄⁺ isotope dilution method in a Danish soil subjected to four different agricultural practices (set aside, barley, winter wheat and clover). Results were compared to arginine ammonification in the soil samples measured as NH₄⁺ production following addition of excess (1 mM) arginine. In the set aside, barley, winter wheat and clover soils the average annual rates of gross N mineralization (0.29, 0.60, 1.34 and 1.75 µg NH₄⁺-N g^-1 day^-1, respectively) and arginine ammonification activity (0.21, 0.55, 0.88, and 1.33 µg NH₄⁺-N g^-1 h^-1, respectively) were well correlated. Furthermore, the seasonal variations of gross N mineralization and arginine ammonification activities were very similar, showing rapid responses to rainfall and generally higher activities in wetted soils. As tested in the laboratory, the arginine ammonification activity correlated well with heterotrophic microbial respiration activity (CO₂ production) in soil samples and further displayed a simple, one-component Michaelis-Menten kinetics with a high affinity for arginine (K_m value of 48 µM LJ µM) as determined from non-linear parameter estimation. This indicated that arginine ammonification activity was primarily due to microorganisms, and the activity was also shown to be at a minimum in sterile soil samples. All evidence thus supported that our standard assay of arginine ammonification activity provides a good index of gross N mineralization rates by the microorganisms in soil under in situ conditions.     

Appetite,metabolism and growth regulation in Atlantic salmon (Salmo salar L.) exposed to hypoxia at elevated seawater temperature

High temperature combined with low dissolved oxygen (DO) is one of the most challenging environmental conditions farmed fish experience; thus, understanding their impact on growth regulation is of relevance to cultured and wild populations. This study examines appetite‐ and growth‐regulating mechanisms in Atlantic salmon postsmolt exposed to either high (HO) or low oxygen (LO) at a suboptimally high temperature (17°C). Additionally, the effects of high (HE) and low (LE) dietary energy (DE) were examined. After a month of treatment, analyses of hormones, regulating appetite (ghrelin) and growth (growth hormone receptor ghr1 and insulin‐like growth factor IGF‐1), and free amino acids (FAA) were measured pre‐ and postprandially at ?4, ?2, 0, 2, 4 and 6 h. No preprandial ghrelin peaks were detected despite a significant reduction in feed intake and growth under hypoxia compared to normoxia. LO treatment also had an overall negative effect on survival compared to HO, while nutrient retention efficiency, FCR and plasma FAA concentrations were unaffected (P > 0.05). Feeding HE diet resulted in increased growth (+17%) and improved FCR (?14%) and energy retention efficiency (+26%) independent of DO. Plasma FAA concentrations were unaffected by LO treatment and DE (P > 0.05). Growth regulatory gene expressions possibly reflect an overall lower growth at a high temperature overriding the impacts of DO and DE. This study also indicates that optimal adaptation time to environmental conditions and feeding regime is crucial for establishing a regular hormonal appetite signalling that reflects real feeding anticipation in salmon.     

Mechanism of Na+/H+ antiporting

Na+/H+ antiporters are central to cellular salt and pH homeostasis. The structure of Escherichia coli NhaA was recently determined, but its mechanisms of transport and pH regulation remain elusive. We performed molecular dynamics simulations of NhaA that, with existing experimental data, enabled us to propose an atomically detailed model of antiporter function. Three conserved aspartates are key to our proposed mechanism: Asp164 (D164) is the Na+-binding site, D163 controls the alternating accessibility of this binding site to the cytoplasm or periplasm, and D133 is crucial for pH regulation. Consistent with experimental stoichiometry, two protons are required to transport a single Na+ ion: D163 protonates to reveal the Na+-binding site to the periplasm, and subsequent protonation of D164 releases Na+. Additional mutagenesis experiments further validated the model.     

Enzymatic production of monoacylglycerols containing polyunsaturated fatty acids through an efficient glycerolysis system

The aim of the study was to develop an efficient glycerolysis system for the enzymatic production of monoacylglycerols (MAGs) containing polyunsaturated fatty acids. Glycerolysis has been widely applied in industry for the chemical production of food MAGs under high temperature. The enzymatic glycerolysis system at 40-70 degrees C is unfortunately a multiphase system, which leads to the lower reaction efficiency. A tert-butyl alcohol system was developed after careful evaluation and more than 20-fold of the reaction efficiency from this system was obtained compared to the solvent-free system. Novozym 435 was employed as a catalyst in the glycerolysis from the screening. In the batch reaction system with tert-butyl alcohol, temperature higher than 40 degrees C was favored. The glycerol/oil ratio was best in the study with 4.5 while the solvent weight ratio from 1 to 3 had little effect. In general, 60-70% yield can be obtained at 2 h in the stirred tank reactor. The continuous glycerolysis was conducted in a packed bed reactor. MAG yield up to 70% was reached at 30-40 min residence time. The continuous glycerolysis was more sensitive to the amount of tert-butyl alcohol, and in the weight ratio to oil more than 2 was favored. The continuous process was optimized with the assistance of response surface methodology. Optimal conditions for the packed bed reactor after all considerations were recommended as glycerol/oil 4:1 (mol/mol), temperature 40 degrees C, and residence time 45 min. The operation stability study showed that there was no slight reduction of reaction performance at more than 30 days, implying a high feasibility in practical applications.     

C and N Turnover and Lignocellulose Degradation During Composting of Miscanthus Straw And Liquid Pig Manure

To obtain a peat substitute for pot plants, Miscanthus straw and liquid pig manure were composted in two different systems (open box and closed reactor) and examined for changes in pH, water content and chemical composition (nitrogen, carbon, hemicellulose, cellulose and lignin). Temperature maxima of 65-70°C were achieved within a few days in both systems. Composting and sampling were continued for 190 days in the box system and 150 days in the reactor. Major loss of nitrogen was found in a period of eight days after temperature maximum followed by stabilization. Accumulation of nitrate was observed during final weeks of composting in both systems. A marked decrease in content of hemicellulose was seen just after temperature maximum followed by slow, constant decrease throughout remaining composting period. The same pattern was observed for degradation of cellulose, though degradation was initiated later in the experiment. Degradation of hemicellulose was nearly 100% in both systems, whereas the cellulose decrease was 64 and 70% in the box and reactor, respectively. Lignin was not degraded during the experiments. Though initial C/N ratio was different in the two systems (25 in box and 16 in reactor), final ratios were the same (C/N=13). In spite of the similarities, the more complicated closed system is preferred if removal of ammonia from exit gas is considered.     

Prediction of Starch,Soluble Sugars and Amino Acids in Potatoes (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Using Hyperspectral Imaging,Dielectric and LF-NMR Methodologies

Handling and processing of potatoes is performed in increasingly large and more automated facilities, and the industry calls for more automated machinery for quality assessment and sorting by concentration of starch, soluble sugars, protein, amino acids etc. of the potato tubers. The present study was designed to evaluate five different scanning methods for their potential use in potato assessment and sorting. Two methods were based on hyperspectral imaging, two were based on dielectric/bio-impedance and one was based on low-field nuclear magnetic resonance. A set of 60 potatoes of 10 different cultivars were simultaneously sampled for analyses of content and scanned by the five different scanning methods. The resulting multivariate dataset was used to estimate the prediction ability of the individual scanning methods on starch-related parameters, selected simple sugars, selected amino acids, conductivity of pressed cell sap and cell sizes. Results showed that most types of spectral analyses had relatively high potential for predicting the starch-related parameters and medium potential for predicting the concentration of the reducing sugars fructose and glucose. Most methods showed medium potential for prediction of several amino acids, including asparagine, which showed particularly promising predictions in the hyperspectral analyses of intact potatoes. The presented screening study enabled us to perform robust choices for the further development and optimization of the methods and instruments for industrial implementation.     

A system for model-based biomass estimation of larvae in intensive cod larvicultures

High and unpredictable mortality rates are observed in the larval rearing of cod (Gadus morhua). As a means of addressing this problem, we present a model-based estimator system which can be used to indirectly measure the larval density through monitoring the live food dynamics and larval growth. The estimator has been evaluated in a conceptual context using a preliminary model formulation, and the observability of the process has been investigated. It was found that the two parameters, live food dynamics and larval growth, contain enough information for the larval density to be estimated under noisy conditions, given the correct model. When the system is applied practically, the estimation error will depend on the measurement and model accuracy; this is especially true with respect to the predictability of the feed intake rate of the fish.     

Lysine limitation alters the storage pattern of protein,lipid and glycogen in on‐growing Atlantic salmon

This experiment aimed to test the interaction of lysine limitation with nutrient accretion and muscle carnitine depot in Atlantic salmon. Fish were fed adequate or low‐lysine diets for 3 months. Lysine intake was significantly less (48%) in fish fed the low‐lysine diet as compared with that fed the adequate one. There was no difference in dietary amino acids between treatments, with the exception of lysine. The lower lysine intake was reflected in plasma free lysine being 52% less while the free lysine concentration in the liver and muscle were unaffected. Although there was no significant difference between voluntary feed intakes among treatments, fish fed the low‐lysine diet had reduced growth, protein and energy deposition as compared with fish fed the adequate lysine diet. White trunk muscle contained more glycogen and less protein in fish fed the low‐lysine diet while no difference in lipid was observed. The livers from fish fed the low‐lysine diet contained less glycogen and slightly more fat and protein than the livers from fish fed the adequate lysine diet. Lysine limitation reduced carnitine in the liver without affecting muscle carnitine depot. Thus, low‐lysine diets did not likely affect the fatty acid oxidation capacity. This fact was supported by unaffected fatty acid profiles and lipid classes between treatments during the 3‐month study. In conclusion, lysine limitation does not deplete the muscle carnitine depot during the on‐growing seawater phase of Atlantic salmon, but affects the deposition pattern of nutrients.     

Evidence of cormorant‐induced mortality,disparate migration strategies and repeatable circadian rhythm in the endangered North Sea houting (Coregonus oxyrinchus): A telemetry study mapping the postspawning migration

Life history theory predicts a trade‐off between migration and residency where migration is favoured when it infers elevated fitness. Although migration to more favourable environments may offer higher growth rates, migrants often experience increased mortality due to predation. Here, we investigated mortality and migration behaviour of the North Sea houting (Coregonus oxyrinchus), an anadromous salmonid endemic to the Wadden Sea. We used acoustic telemetry to map the migration of the only remaining indigenous population by applying stationary hydrophones combined with manual tracking. Data suggested a total mortality of 26%, with 30% of the total mortality attributed to predation by great cormorants (Phalacrocorax carbo sinensis), highlighting that North Sea houting conservation could be jeopardised by increased cormorant predation. Risk of cormorant predation was size‐dependent, with smaller fish suffering higher risk of predation. The study found North Sea houting to exhibit disparate migration strategies and identified a lentic area in the estuary as an important habitat. Two newly established artificial lakes within the river system significantly reduced the migration speeds, possibly indicating constrained navigation through the lakes. The migration into the Wadden Sea correlated with temperature perhaps indicating osmoregulatory constraints of sea entry. Unlike many salmonid species, migration occurred both day and night. Moreover, fish exhibited repeatable individual differences in diel activity patterns, suggesting that individuals differ consistently in their migratory activity throughout the 24‐hr period. Our study provides novel information on salmonid migration, which is crucial for the development of science‐based conservation strategies.