Ammonia Accumulation Research Articles

Impaired enzymatic antioxidant defense in erythrocytes of rats with ammonia-induced encephalopathy: role NMDA receptors

Hepatic encephalopathy (HE), a medical term that deals with a neuropsychiatric disorder seen in patients with severe hepatic dysfunction, has been known for more than a century, however, pathogenetic mechanisms underlying cerebral dysfunction during conditions of liver disease are still not entirely understood. There is a consensus that an accumulation of ammonia in brain as a result of impaired detoxification capacity of the liver or the appearance of a portosystemic shunt is a primary cause of HE. Current evidence suggests that ammonia toxicity is mediated by hyperactivation of glutamate receptors, mainly N-methyl-D-aspartate receptors (NMDA-R) and affects many processes of aerobic metabolism that provides energy for a myriad of specific functions and viability of nerve cells. Recent reports on the presence of functional NMDA receptors in erythrocytes and deviations from normal ranges in many blood test parameters that was indicative of impaired hemodynamics and reduced carrying capacity for oxygen in erythrocytes in most patients with HE implicate the relationship between “erythrocyte damage” and cerebral dysfunction. In order to understand how, during hyperammonemia, disturbances in energy metabolism in brain that needs a constant supply of a high level of oxygen in the blood lead to encephalopathy, it is necessary to reveal ammonia-induced disorders in energy metabolism and antioxidant defense system of erythrocytes and explore the potential role of ammonia in reduced brain oxygenation. To detect the said missing link, the activities of erythrocyte antioxidant enzymes and concentrations of GSH, GSSG and H2O2 were measured in animals with hyperammonemia by using MK-801, a powerful noncompetitive NMDA receptor antagonist. It was found that the accumulation of ammonia in the blood of animals with hyperammonemia occurs within rat erythrocytes in large amounts and makes these cells which do not contain enzymes for the removal of ammonia, more susceptible to a prooxidant environment of ammonia created during oxidative stress that was completely or partially inhibited with the help of MK-801. Data obtained provide a support to identification of extra risk factors in cognitive disorders and in the prediction of unfavorable outcome with hypoperfusion in patients with elevated concentrations of ammonia in the blood.

Imbalanced metabolism induced NH4+ accumulation and its effect on the central metabolism of Methylomonas sp. ZR1.

Nitrogen and carbon are the two most essential nutrient elements, and their metabolism is tightly coupled in single carbon metabolic microorganisms. However, the nitrogen metabolism and the nitrogen/carbon (N/C) metabolic balance in single-carbon metabolism is poorly studied. In this study, the nitrogen metabolism pattern of the fast growing methanotrophs Methylomonas sp. ZR1 grown in methane and methanol was studied. Effect study of different nitrogen sources on the cell growth of ZR1 indicates that nitrate salts are the best nitrogen source supporting the growth of ZR1 using methane and methanol as carbon source. However, its metabolic intermediate ammonium was found to accumulate with high N/C ratio in the medium and consequently inhibit the growth of ZR1. Studies of carbon and nitrogen metabolic kinetic under different N/C ratio conditions indicate that the accumulation of NH4+ is caused by the imbalanced nitrogen and carbon metabolism in ZR1. Feeding carbon skeleton α-ketoglutaric acid could effectively relieve the inhibition effect of NH4+ on the growth of ZR1, which further confirms this assumption. qPCR analysis of the expression level of the central metabolic key enzyme gene indicates that the nitrogen metabolic intermediate ammonium has strong regulation effect on the central nitrogen and carbon metabolism in ZR1. qPCR-combined genomic analysis confirms that a third ammonium assimilation pathway glycine synthesis system is operated in ZR1 to balance the nitrogen and carbon metabolism. Based on the qPCR result, it was also found that ZR1 employs two strategies to relieve ammonium stress in the presence of ammonium: assimilating excess ammonium or cutting off the nitrogen reduction reactions according to the available C1 substrate. Validating the connections between single-carbon and nitrogen metabolism and studying the accumulation and assimilation mechanism of ammonium will contribute to understand how nitrogen regulates cellular growth in single-carbon metabolic microorganisms.

Minimal hepatic encephalopathy: clinical, neurophysiological, neuroimaging markers

Hepatic encephalopathy in the initial stages is a diagnostically challenging clinical phenomenon, based on the accumulation of ammonia. Symptoms of encephalopathy are extremely varied: from a slight cognitive decrease and minimal affective disorders to disorders of consciousness and coma in the terminal stages. However, the severity of liver pathology and neurophysiological and neuroimaging data do not always correlate with the severity of encephalopathy. The greatest difficulties in diagnosis arise at the initial stage of the disease, and a timely recognized and established diagnosis can not only slow down the progression of cognitive deficits and characterological changes, but also significantly improve the patient’s prognosis.

A perspective on Bio-Electrochemical System (BES) as a tool for boosting the performance of the nonperforming anaerobic units

Anaerobic technologies (AT) offer a sustainable approach for harnessing energy during wastewater treatment. Despite their numerous advantages, significant challenges remain. Under stressful conditions, the accumulation of ammonia and Volatile Fatty Acids (VFA) within AT can lead to a decline in its performance. One promising strategy to enhance the performance of sick anaerobic units is the introduction of Syntrophic Fatty Acid Oxidizing Bacteria (SFAOB), reinforcing the crucial partnership between SFAOB and hydrogenotrophic methanogenic (HM) organisms. However, isolating and enriching SFAOB populations is challenging due to thermodynamic limitations. To address this issue, Bio-electrochemical systems (BES) emerge as a valuable tool for SFAOB enrichment. BES can also play a vital role in removing dissolved ammonium from AT, preventing its toxic build-up within the system. This perspective primarily focuses on the role of BES in augmenting the performance of AT. Employing BES as a performance-enhancing solution holds promise for ensuring the sustainable recovery of impaired anaerobic units.

Raman-controlled pyruvate feeding to control metabolic activity and product quality in continuous biomanufacturing.

Despite technological advances ensuring stable cell culture perfusion operation over prolonged time, reaching a cellular steady-state metabolism remains a challenge for certain manufacturing cell lines. This study investigated the stabilization of a steady-state perfusion process producing a bispecific antibody with drifting product quality attributes, caused by shifting metabolic activity in the cell culture. A novel on-demand pyruvate feeding strategy was developed, leveraging lactate as an indicator for tricarboxylic acid (TCA) cycle saturation. Real-time lactate monitoring was achieved through in-line Raman spectroscopy, enabling accurate control at predefined target setpoints. The implemented feedback control strategy resulted in a three-fold reduction of ammonium accumulation and stabilized product quality profiles. Stable and flat glycosylation profiles were achieved with standard deviations below 0.2% for high mannose and fucosylation. Whereas galactosylation and sialylation were stabilized in a similar manner, varying lactate setpoints might allow for fine-tuning of these glycan forms. The Raman-controlled pyruvate feeding strategy represents a valuable tool for continuous manufacturing, stabilizing metabolic activity, and preventing product quality drifting in perfusion cell cultures. Additionally, this approach effectively reduced high mannose, helping to mitigate increases associated with process intensification, such as extended culture durations or elevated culture densities.

Flaxseed Oil (Linum usitatissimum) Prevents Cognitive and Motor Damage in Rats with Hyperammonemia.

Hyperammonemia is characterized by the excessive accumulation of ammonia in the body as a result of the loss of liver detoxification, leading to the development of hepatic encephalopathy (HE). These metabolic alterations carry cognitive and motor deficits and cause neuronal damage, with no effective treatment at present. In this study, we aimed to evaluate the effect of two subacute oral administrations of flaxseed oil (0.26 and 0.52 mL/kg) on short- and long-term memory, visuospatial memory, locomotor activity, motor coordination, and the neuronal morphology of the prefrontal cortex (PFC) via tests on Wistar rats with hyperammonemia. The goal was to identify its role in the regulation of cerebral edema, without liver damage causing cerebral failure. In contrast with an ammonium-rich diet, flaxseed oil and normal foods did not cause cognitive impairment or motor alterations, as evidenced in the short-term and visuospatial memory tests. Furthermore, the flaxseed oil treatment maintained a regular neuronal morphology of the prefrontal cortex, which represents a neuroprotective effect. We conclude that the oral administration of flaxseed oil prevents cognitive and motor impairments as well as neuronal alterations in rats with hyperammonemia, which supports the potential use of this oil to ameliorate the changes that occur in hepatic encephalopathy.

Dissolved organic matter and nutrient processing in organic-rich subterranean estuaries: Implications for future land use and climate scenarios

The subterranean estuary (STE) is a recognized reactive biogeochemical zone at the groundwater–seawater interface in coastal regions. However, few studies have explored dissolved organic matter (DOM) and nutrient processes in buried organic-rich STEs (with up to 20 % of organic matter content), despite their potential contributions to coastal water hypoxia and greenhouse gas emissions. This study conducted controlled laboratory experiments to investigate the physicochemical controls on DOM and nutrient processing in organic-rich STEs, considering variations in groundwater nitrate (NO3–) concentrations, salt concentration, seawater ions, and incubation times. Results showed elevated levels of dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) in pore water, primarily from particulate organic matter (POM) degradation that originated from decayed vascular plants. POM-to-DOM degradations further resulted in the accumulation of ammonium (NH4+) and highly unsaturated compounds in all treatments. The addition of high-ionic-strength treatments (e.g., seawater and sodium chloride [NaCl] solution) reduced DOC solubility and selectively removed dissolved aromatic compounds in porewater. The cation-rich seawater shifted the porewater NH4+ generation pathway from predominantly biological (re)mineralization in freshwater to abiotic (desorption via ion exchange) pathways in the saline water setup. Orthophosphate (PO43−) dynamics were likely to be influenced by the iron-rich sediment surface in both freshwater and saline water scenarios. Our experiments further indicated that the long-term incubation enriched DOM concentration with a high degradation state. This work demonstrated that organic-rich STEs are significant sources of less labile DOM and NH4+ to coastal waters. In the context of global climate change, our results illustrate the sensitivity of DOM and nutrient biogeochemical cycling in STEs to salinization and altered residence time, consequently affecting the quantity and quality of DOM and nutrients transported to coastal waters.

Peptonics: A new family of cell-protecting surfactants for the recombinant expression of therapeutic proteins in mammalian cell cultures.

Polymer surfactants are key components of cell culture media as they prevent mechanical damage during fermentation in stirred bioreactors. Among cell-protecting surfactants, Pluronics are widely utilized in biomanufacturing to ensure high cell viability and productivity. Monodispersity of monomer sequence and length is critical for the effectiveness of Pluronics-since minor deviations can damage the cells-but is challenging to achieve due to the stochastic nature of polymerization. Responding to this challenge, this study introduces Peptonics, a novel family of peptide and peptoid surfactants whose monomer composition and sequence are designed to achieve high cell viability and productivity at a fraction of chain length and cost of Pluronics. A designed ensemble of Peptonics was initially characterized via light scattering and tensiometry to select sequences whose phase behavior and tensioactivity align with those of Pluronics. Selected sequences were evaluated as cell-protecting surfactants using Chinese hamster ovary (CHO) cells expressing therapeutic monoclonal antibodies (mAb). Peptonics IH-T1010, ih-T1010, and ih-T1020 afforded high cell density (up to 3×107 cellsmL-1 ) and viability (up to 95% within 10 days of culture), while reducing the accumulation of ammonia (a toxic metabolite) by ≈10% compared to Pluronic F-68. Improved cell viability afforded high mAb titer (up to 5.5mgmL-1 ) and extended the production window beyond 14 days; notably, Peptonic IH-T1020 decreased mAb fragmentation and aggregation ≈5%, and lowered the titer of host cell proteins by 16% compared to Pluronic F-68. These features can improve significantly the purification of mAbs, thus increasing their availability at a lower cost to patients.

The malate shuttle detoxifies ammonia in exhausted T cells by producing 2-ketoglutarate

The malate shuttle is traditionally understood to maintain NAD+/NADH balance between the cytosol and mitochondria. Whether the malate shuttle has additional functions is unclear. Here we show that chronic viral infections induce CD8+ T cell expression of GOT1, a central enzyme in the malate shuttle. Got1 deficiency decreased the NAD+/NADH ratio and limited antiviral CD8+ T cell responses to chronic infection; however, increasing the NAD+/NADH ratio did not restore T cell responses. Got1 deficiency reduced the production of the ammonia scavenger 2-ketoglutarate (2-KG) from glutaminolysis and led to a toxic accumulation of ammonia in CD8+ T cells. Supplementation with 2-KG assimilated and detoxified ammonia in Got1-deficient T cells and restored antiviral responses. These data indicate that the major function of the malate shuttle in CD8+ T cells is not to maintain the NAD+/NADH balance but rather to detoxify ammonia and enable sustainable ammonia-neutral glutamine catabolism in CD8+ T cells during chronic infection.

Extreme accumulation of ammonia on electroreduced mackinawite: An abiotic ammonia storage mechanism in early ocean hydrothermal systems.

An increasing amount of evidence suggests that early ocean hydrothermal systems were sustained sources of ammonia, an essential nitrogen species for prebiotic synthesis of life's building blocks. However, it remains a riddle how the abiotically generated ammonia was retained at the vent-ocean interface for the subsequent chemical evolution. Here, we demonstrate that, under simulated geoelectrochemical conditions in early ocean hydrothermal systems ([Formula: see text][Formula: see text] V versus the standard hydrogen electrode), mackinawite gradually reduces to zero-valent iron ([Formula: see text]), generating interlayer [Formula: see text] sites. This reductive conversion leads to an up to 55-fold increase in the solid/liquid partition coefficient for ammonia, enabling over 90% adsorption of 1 mM ammonia in 1 M NaCl at neutral pH. A coordinative binding of ammonia on the interlayer [Formula: see text] sites was computed to be the major mechanism of selective ammonia adsorption. Mackinawite is a ubiquitous sulfide precipitate in submarine hydrothermal systems. Given its reported catalytic function in amination, the extreme accumulation of ammonia on electroreduced mackinawite should have been a crucial initial step for prebiotic nitrogen assimilation, paving the way to the origin of life.

The effect of soil salt content and ionic composition on nitrification in a Fluvisol of the Yellow River Delta

Soil salinization is a common cause of land degradation and a major constraint on global food production. However, the effects that different salts have on soil biogeochemical processes such as nitrification are not fully understood. An incubation experiment was conducted with two different salt treatments applied at varying concentrations (sodium chloride addition, NaCl1: 2.06, NaCl2: 4.12, NaCl3: 5.00 g kg−1; sodium sulphate addition, NaS1: 5.00, NaS2: 6.07, NaS3:12.14 g kg−1). A Fluvisol soil was used as incubation medium and urea was supplied as the nitrogen source. Results showed that urease activities were reduced (28–76%) in salt treated soils on day 1, but recovered over time and finally surpassed that of the control on day 56. Moreover, the average urease activity of Na2SO4 treatments was 1.26 times higher than that of NaCl treatments on day 56. In addition, increasing soil salinity levels simultaneously reduced hydrolysis and nitrification rates, which led to the temporary accumulation of ammonium (NH4+-N) in soils for up to 14 days. The maximum contents of NH4+-N accumulation in NaCl3 and NaS3 were 1.64 and 1.48 times higher than those of NaCl1 and NaS1, respectively. Nitrification rates were inhibited more severely in soils dominated by Cl-. Low soil salt contents (less than 7.79 g kg−1) only delayed nitrification without reducing the total amount of nitrate (NO3--N) produced during the incubation. However, high salt contents (NaS3) significantly decreased the total NO3--N production. Our results indicate that nitrite oxidation process may also limit the speed of nitrification in salt-affected soils. Additionally, a new model is introduced to better simulate the dynamics of nitrification in salt-affected soils. In conclusion, soil salt content is the primary factor that inhibits nitrification. Anions affect the inhibitory strength of salinity on nitrification in soils with similar salinity levels, where Cl- is more suppressive than SO42-.

The heterogeneity of microbial diversity and its drivers in two types of sediments from tidal flats in Beibu Gulf, China

Mudflats and sandflats are two common types of coastal tidal flats, the structure and function differences of microbial communities between them are still underappreciated. Beibu Gulf is a diurnal tidal regime located in China, the differences between the two type of tidal flats could be more distinct. In this study, we collected a total of 6 samples from Beibu Gulf, consisting of 3 sandflats samples and 3 mudflats samples, classified based on clay and silt content. Generally, the mudflats samples exhibited higher levels of NH4+_N and TOC, but lower in ORP and pH. The microbial diversity of the two types of tidal flats was investigated, revealing great differences existed and sandflats had higher microbial richness and diversity than mudflats. Furthermore, we analyzed the association between microbial communities and environmental factors, finding NH4+_N to have the highest contribution to the total variation in microbial community structure, and microbial groups such as Desulfobacterota, Campilobacterota, Chloroflexota, Calditrichota, Spirochaetota, Zixibacteria, Latescibacterota and Sva0485 group in mudflats were positively associated with NH4+_N. The functions of microbial community were predicted using metagenomic sequences and metagenome assembled genome (MAG). Mudflats contained more genes for carbon fixation. Nitrate and nitrite reduction were widely existed in mudflats and sandflats, but nitrogen fixation was only existed in mudflats, and Campilobacterota, Desulfobacterota and Gammaproteobacteria MAGs were mainly responsible for it. Sandflats composed more genes for ammonium oxidation, but no MAG was found whether in sandflats or mudflats. Microbes in mudflats exhibited a greater abundance of genes related to sulfur cycling, especially in reduction process, unique MAGs in mudflats such as Calditrichota, Chloroflexota, Desulfobacterota and Zixibacteria MAGs are responsible for sulfate and sulfite reduction. Finally, we predicted functions of ammonium related microbes in mudflats based on MAGs and found Campilobacterota and Desulfobacterota MAGs were important for high accumulation of ammonium in mudflats. This study illuminated the structural and functional differences of microbial communities in mudflats and sandflats, providing new insights into the relationship of microbial communities and environment in the tidal flat.

Effects of manipulating lactate dehydrogenase gene on metabolism of HEK-293 and production of human adenovirus

Reducing lactate accumulation has always been a goal of the mammalian cell biotechnology industry. When animal cells are cultured in vitro, the accumulation of lactate is mainly the combined result of two metabolic pathways. On one hand, glucose generates lactate under the function of lactate dehydrogenase A (LDHA); on the other hand, lactate can be oxidized to pyruvate by LDHB or LDHC and re-enter the TCA cycle. This study comprehensively evaluated the effects of LDH manipulation on the growth, metabolism and human adenovirus (HAdV) production of human embryonic kidney 293 (HEK-293) cells, providing a theoretical basis for engineering the lactate metabolism in mammalian cells. By knocking out ldha gene and overexpression of ldhb and ldhc genes, the metabolic efficiency of HEK-293 cells was effectively improved, and HAdV production was significantly increased. Compared with the control cell, LDH manipulation promoted cell growth, reduced the accumulation of lactate and ammonia, significantly enhanced the efficiency of substrate and energy metabolism of cells, and significantly increased the HAdV production capacity of HEK-293 cells. Among these LDH manipulation measures, ldhc gene overexpression performed the best, with the maximum cell density increased by about 38.7%. The yield of lactate to glucose and ammonia to glutamine decreased by 33.8% and 63.3%, respectively; and HAdV titer increased by at least 16 times. In addition, the ATP production rate, ATP/O2 ratio, ATP/ADP ratio and NADH content of the modified cell lines were increased to varying degrees, and the energy metabolic efficiency was significantly improved.

Combination of tea catechins and ornithine effectively activates the urea cycle: an in vitro and human pilot study.

Accumulation of ammonia causes central and peripheral fatigue. This study aimed to investigate the synergistic effect of tea catechins and low-dose ornithine in activating the urea cycle to reduce blood ammonia levels during exercise. We used hepatocyte-like cells derived from human-induced pluripotent stem (iPS) cells to assess the effect of tea catechins combined with ornithine on urea cycle activity. The urea production and expression of key genes involved in the metabolism of urea were investigated. We then examined the synergistic improvement in ammonia metabolism by tea catechins in combination with ornithine in a human pilot study. Tea catechins combined with ornithine increased urea cycle activity in hepatocyte-like cells derived from human iPS cells. Intake of 538.6mg of tea catechins with 1592mg of ornithine for 2 consecutive days during exercise loading suppressed the exercise-induced increase in the blood ammonia concentration as well as stabilized blood glucose levels. Controlling the levels of ammonia, a toxic waste produced in the body, is important in a variety of situations, including exercise. The present study suggests that a heterogeneous combination of polyphenols and amino acids efficiently suppresses elevated ammonia during exercise in humans by a mechanism that includes urea cycle activation. This study was registered in the University Hospital Medical Information Network Clinical Trial Registry (No. UMIN000035484, dated January 8, 2019).

Impaired Enzymatic Antioxidant Defense in Erythrocytes of Rats with Ammonia-Induced Encephalopathy: Role of NMDA Receptors.

Hepatic encephalopathy (HE), a neuropsychiatric disorder developing in patients with severe hepatic dysfunction, has been known for more than a century. However, pathogenetic mechanisms of cerebral dysfunction associated with liver disease are still poorly understood. There is a consensus that the primary cause of HE is accumulation of ammonia in the brain as a result of impaired liver detoxification capacity or the portosystemic shunt. Current evidence suggests that ammonia toxicity is mediated by hyperactivation of glutamate receptors, mainly N-methyl-D-aspartate receptors (NMDARs), and affects brain aerobic metabolism, which provides energy for multiple specific functions and neuronal viability. Recent reports on the presence of functional NMDARs in erythrocytes and the data on the deviations of blood parameters from their normal ranges indicate impaired hemodynamics and reduced oxygen-carrying capacity of erythrocytes in most patients with HE, thus suggesting a relationship between erythrocyte damage and cerebral dysfunction. In order to understand how hyperammonemia (HA)-induced disturbances in the energy metabolism in the brain (which needs a constant supply of large amounts of oxygen in the blood) lead to encephalopathy, it is necessary to reveal ammonia-induced impairments in the energy metabolism and antioxidant defense system of erythrocytes and to explore a potential role of ammonia in reduced brain oxygenation. To identify the said missing link, the activities of antioxidant enzymes and concentrations of reduced glutathione (GSH), oxidized glutathione (GSSG), and H2O2 were measured in the erythrocytes of rats with HA that were injected with the noncompetitive NMDAR antagonist MK-801. We found that in rats with HA, ammonia was accumulated in erythrocytes (cells lacking ammonia removal enzymes), which made them more susceptible to the prooxidant environment created during oxidative stress. This effect was completely or partially inhibited by MK-801. The data obtained might help to identify the risk factors in cognitive disorders and facilitate prediction of unfavorable outcomes of hypoperfusion in patients with a blood elevated ammonia concentration.

Lethality rescue and long-term amelioration of a citrullinemia type I mouse model by neonatal gene-targeting combined to SaCRISPR-Cas9

Citrullinemia type I is a rare autosomal-recessive disorder caused by deficiency of argininosuccinate synthetase (ASS1). The clinical presentation includes the acute neonatal form, characterized by ammonia and citrulline accumulation in blood, which may lead to encephalopathy, coma, and death, and the milder late-onset form. Current treatments are unsatisfactory, and the only curative treatment is liver transplantation. We permanently modified the hepatocyte genome in lethal citrullinemia mice (Ass1fold/fold) by inserting the ASS1 cDNA into the albumin locus through the delivery of two AAV8 vectors carrying the donor DNA and the CRISPR-Cas9 platform. The neonatal treatment completely rescued mortality ensuring survival up to 5months of age, with plasma citrulline levels significantly decreased, while plasma ammonia levels remained unchanged. In contrast, neonatal treatment with a liver-directed non-integrative AAV8-AAT-hASS1 vector failed to improve disease parameters. To model late-onset citrullinemia, we dosed postnatal day (P) 30 juvenile animals using the integrative approach, resulting in lifespan improvement and a minor reduction in disease markers. Conversely, treatment with the non-integrative vector completely rescued mortality, reducing plasma ammonia and citrulline to wild-type values. In summary, the integrative approach in neonates is effective, although further improvements are required to fully correct the phenotype. Non-integrative gene therapy application to juvenile mice ensures a stable and very efficient therapeutic effect.

Efficacy of isobutanol to reduce transport stress of juvenile ornate tropical rock lobster (TRL; Panulirus ornatus)

The ornate tropical rock lobster (TRL; Panulirus ornatus) is a fast growing spiny lobster that is prized for its vivid colouration and excellent eating qualities. The TRL aquaculture industry is reliant on the successful transport of juveniles to often distant grow-out sites from where they are harvested in the wild or raised in hatcheries. There has been mixed success transporting TRL juveniles, with high mortality rates which are typically associated with metabolic stress. Isobutanol, a food safe alcohol, has been shown to act as a sedative in adult TRL, as well as several other commercially important lobster species. This study found that juvenile TRL were rapidly sedated with isobutanol treatments at 350 and 500 ppm (15.05 and 6.47 min, respectively) and sustained inactivity was observed (36.12 and 60.62 min, respectively). Pre-treatment with isobutanol at 25 and 50 ppm led to decreased CO2 production rates during a 2 h air exposure experiment compared to untreated individuals (∼55 and 28%, respectively). Glycogen stores were maintained (∼75 higher) while lactate and ammonia concentrations were lower (∼50% and 40%, respectively) compared to untreated individuals. Isobutanol treatments at 25 and 50 ppm prior to 20 h of simulated transport improved overall survival (87.5% without treatment; 100% at either isobutanol treatment), while maintaining glycogen stores (∼80% higher) and limiting the accumulation of lactate and ammonia (∼25% and ∼ 45% lower, respectively) compared with untreated individuals. The results indicate that isobutanol treatment prior to air exposure during transport has the potential to improve outcomes from the transport of live juvenile TRL by decreasing overall metabolic stress and increasing survival.

Biogeochemical and physical controls on ammonium accumulation on the Chukchi shelf, western Arctic Ocean

Spatial variability of ammonium concentrations along repeat transects were examined on the Chukchi shelf during 2012–2018. Two distinct near-bottom high ammonium pools (>1 μmol/kg) near 67.5°N and 72.5°N of the transects were identified in all years. The accumulation of ammonium in the regions is driven primarily by a combination of biogeochemical processes (e.g., dynamic bacterial remineralization of organic matter) and physical controls (e.g., strong density-contrast barrier limits upward mixing of ammonium). The ammonium pool on the shelf may became larger in the expectation of the stronger bacterial remineralization following elevate primary production, and may have potential impact on the structure and productivity of ecosystem on the Chukchi shelf.

Simulated transport of flathead grey mullet Mugil cephalus eggs: Effects of density and duration

Flathead grey mullet has a great potential as a low trophic level aquaculture species, however captive rearing is still hampered by the lack of well-established production protocols. Here we examined the effects of transport time and egg loading density on water quality, egg survival, hatching success and early larvae performance when shipped for 8, 16 and 24 h at 500 and 1500 eggs L−1. When no water conditioners were used, survival immediately after transport was consistently over 60% up to 16 h shipment for both densities tested, with minimum values of 40% for 1500 eggs L−1. Density and transport time did not influence hatching rate (p > .05) which ranged between 31 and 54% after 8 and 16 hour transport. However, manipulation after 8 and 24 h transport led to a significant drop in early larvae survival regardless the density tested (20% and 5% respectively) compared to 16 h transport (40%) (p < .05). Hence, the results indicate that grey mullet is sensitive when manipulated as early hatched larvae and when eggs undergo blastophore closure at circa 20 h post-fertilization.The tested egg stocking densities do not deteriorate water quality to lethal levels in terms of pH, NO2 and NO3 especially for short and medium transportation times, however, 24 h transportation induces an accumulation of ammonia affecting the long-term survival of larvae. When basic technology is available (i.e. no water conditioners, refrigerated vehicles), we suggest manipulating grey mullet eggs at 28 h post fertilization for best results in terms of hatching rate and early larvae survival.

Pengaruh Perbedaan Jenis dan Konsentrasi Sumber Nitrogen (NaNO3 dan urea) terhadap Produksi Biomasa Spirulina Platensis

The goal of this study was to investigate the effect of the concentration and the type of nitrogen sources in the cultivation medium on the production of biomass and biochemical content (carbohydrates, proteins, fats) in the Spirulina platensis biomass. Types of nitrogen sources included NaNO3 and urea, where the concentrations were varied from 0.5 to 3.5 g/L. The results showed that the use of urea tended to produce more S. platensis biomass than the use of NaNO3. The best concentrations that produced the highest biomass were NaNO3 2.5 g/L (biomass 0.6745 g/L) and urea 0.5 g/L (biomass 0.7158 g/L). The use of urea also tended to produce a higher specific growth rate and a shorter doubling time than the use of NaNO3. The high concentration of nitrogen sources caused the pH of the medium to increase more rapidly due to the accumulation of ammonium in the medium. Using NaNO3 can produce biomass with a higher protein content (28.34-36.98%) than using urea (25.86-33.52%).