Ammonia Accumulation Research Articles

A glutamine metabolic switch supports erythropoiesis.

Metabolic requirements vary during development, and our understanding of how metabolic activity influences cell specialization is incomplete. Here, we describe a switch from glutamine catabolism to synthesis required for erythroid cell maturation. Glutamine synthetase (GS), one of the oldest functioning genes in evolution, is activated during erythroid maturation to detoxify ammonium generated from heme biosynthesis, which is up-regulated to support hemoglobin production. Loss of GS in mouse erythroid precursors caused ammonium accumulation and oxidative stress, impairing erythroid maturation and recovery from anemia. In β-thalassemia, GS activity is inhibited by protein oxidation, leading to glutamate and ammonium accumulation, whereas enhancing GS activity alleviates the metabolic and pathological defects. Our findings identify an evolutionarily conserved metabolic adaptation that could potentially be leveraged to treat common red blood cell disorders.

Gill-associated ammonia oxidizers are widespread in teleost fish.

Recent advances in sequencing methods have greatly expanded the knowledge of teleost-associated microorganisms. While fish-gut microbiomes are comparatively well studied, less attention has gone toward other, external organ-microbiome associations. Gills are particularly interesting to investigate due to their functions in gas exchange, osmoregulation, and nitrogen excretion. We recently discovered a branchial symbiosis between nitrogen-cycling bacteria and teleosts (zebrafish and carp), in which ammonia-oxidizing Nitrosomonas and denitrifying bacteria together convert toxic ammonia excreted by the fish into harmless dinitrogen (N2) gas. This symbiosis can function as a "natural biofilter" in fish gills and can potentially occur in all ammonotelic fish species, but it remains unknown how widespread this symbiosis is. In this study, we analyzed all publicly available gill microbiome data sets and checked for the presence of Nitrosomonas. We discovered that more than half of the described fish gill microbiomes contain 16S rRNA gene sequences of ammonia-oxidizing bacteria (AOB). The presence of gill-specific AOB was shown in both wild and aquacultured fish, as well as in marine and freshwater fish species. Based on these findings, we propose that ammonia oxidizers are widespread in teleost fish gills. These gill-associated AOB can significantly affect fish nitrogen excretion, and the widespread nature of this association suggests that the gill-associated AOB can have similar impacts on more fish species. Future research should address the contribution of these microorganisms to fish nitrogen metabolism and the fundamental characteristics of this novel symbiosis.IMPORTANCERecent advances in sequencing have increased our knowledge of teleost-associated microbiota, but the gill microbiome has received comparatively little attention. We recently discovered a consortium of nitrogen-cycling bacteria in the gills of common carp and zebrafish, which are able to convert (toxic) ammonia into harmless dinitrogen gas. These microorganisms thus function as a natural nitrogen biofilter. We analyzed all available gill microbiome data sets to determine how widespread gill-associated ammonia-oxidizing bacteria (AOB) are. More than half of the data sets contained AOB, representing both aquacultured and wild fish from freshwater and marine habitats. In total, 182 amplicon sequencing variants were obtained, of which 115 were found specifically in the gills and not the environmental microbiomes. As gill-associated AOB are apparently widespread in teleost fish, it is important to study their impact on host nitrogen excretion and the potential to reduce ammonia accumulation in (recirculating) aquaculture of relevant fish species.

Freshness/spoilage monitoring of protein-rich foods by betacyanin-loaded gelatin/carboxymethyl cellulose halochromic labels

In the present investigation, easy-to-use colorimetric on-package labels were developed based on gelatin/carboxymethyl cellulose films loaded with betacyanin-rich Mirabilis jalapa extract (MJE) for freshness tracking of shrimp and minced meat. The effect of MJE addition (2 %, 4 %, and 6 % (w/v)) on the morphology, micro-structural, optical, and functional characteristics of the labels was assessed. The addition of MJE indicated dose-dependent improvement of antioxidant potential and light barrier capability of the labels but the reduction of their mechanical strength and water barrier properties. The labels underwent color changes (pinkish-red to yellow) after being exposed to different pHs (2–13), which was similar to the MJE solutions when immersed in buffer solutions. The halochromic label loaded with 6 % (w/v) MJE exhibited the highest sensitivity to ammonia. This label was also capable of detecting ammonia concentrations ranging from 0.01 to 0.1 mg/mL, with a visible color shift from red to pale yellow. When utilized for freshness monitoring of shrimp and minced meat, this label displayed noticeable color changes resulting from the accumulation of volatile ammonia and changes in pH during the spoilage of these samples. The labels were also biodegradable, disintegrating within ∼21 days to a great extent. As a result, we found that a label containing 6 % (w/v) MJE can be used as a biodegradable halochromic indicator in the food packaging industry for visual quality monitoring of seafood and meat products.

Positive effects of appropriate micro-aeration on landfill stabilization: Mitigating ammonia and VFAs accumulation

The slow stabilization process of landfill had brought obstacles to urbanization. The paper investigated the efficacy and mechanism of micro-aeration intensity for landfill stabilization. The micro-aeration intensity of 0.05 L/(h·kg) resulted in a significant increase of volatile fatty acids (VFAs) in the hydrolysis stage, and the NH4+-N concentration was reduced by 22.1 %. At the end of landfill, VFAs were rapidly degraded and organic matter was reduced from 36 % to 16 %, which was 55.5 % more efficient than the control group. In addition, the community succession and structure of bacteria and archaea were analyzed. The micro-aeration intensity of 0.05 L/(h·kg) increased the abundance of hydrolyzing functional bacteria such as Pseudomonas and Bacillus, and allowed methanogenic bacteria such as Methanobacterium and Methanothrix to gradually establish oxygen tolerance in the microaerobic environment. The appropriate micro-aeration intensity can accelerate the stabilization process of landfill, which has environmental and economic benefits.

Ammonia-induced lysosomal and mitochondrial damage causes cell death of effector CD8+ T cells.

Ammonia is thought to be a cytotoxin and its increase in the blood impairs cell function. However, whether and how this toxin triggers cell death under pathophysiological conditions remains unclear. Here we show that ammonia induces a distinct form of cell death in effector T cells. We found that rapidly proliferating T cells use glutaminolysis to release ammonia in the mitochondria, which is then translocated to and stored in the lysosomes. Excessive ammonia accumulation increases lysosomal pH and results in the termination of lysosomal ammonia storage and ammonia reflux into mitochondria, leading to mitochondrial damage and cell death, which is characterized by lysosomal alkalization, mitochondrial swelling and impaired autophagic flux. Inhibition of glutaminolysis or blocking lysosomal alkalization prevents ammonia-induced T cell death and improves T cell-based antitumour immunotherapy. These findings identify a distinct form of cell death that differs from previously known mechanisms.

Ammonia-induced stress response in liver disease progression and hepatic encephalopathy.

Ammonia levels are orchestrated by a series of complex interrelated pathways in which the urea cycle has a central role. Liver dysfunction leads to an accumulation of ammonia, which is toxic and is strongly associated with disruption of potassium homeostasis, mitochondrial dysfunction, oxidative stress, inflammation, hypoxaemia and dysregulation of neurotransmission. Hyperammonaemia is a hallmark of hepatic encephalopathy and has been strongly associated with liver-related outcomes in patients with cirrhosis and liver failure. In addition to the established role of ammonia as a neurotoxin in the pathogenesis of hepatic encephalopathy, an increasing number of studies suggest that it can lead to hepatic fibrosis progression, sarcopenia, immune dysfunction and cancer. However, elevated systemic ammonia levels are uncommon in patients with metabolic dysfunction-associated steatotic liver disease. A clear causal relationship between ammonia-induced immune dysfunction and risk of infection has not yet been definitively proven. In this Review, we discuss the mechanisms by which ammonia produces its diverse deleterious effects and their clinical relevance in liver diseases, the importance of measuring ammonia levels for the diagnosis of hepatic encephalopathy, the prognosis of patients with cirrhosis and liver failure, and how our knowledge of inter-organ ammonia metabolism is leading to the development of novel therapeutic approaches.

Adenine nucleotide content and activity of AMP catabolism enzymes in the kidney of rats fed on diets with different protein and sucrose content

Background. Excessive consumption of sucrose or protein deficiency in the diet can induce metabolic disorders in the kidney, whose functioning requires significant ATP energy expenditure. The study investigated the levels of the purine nucleotides ATP, ADP, AMP, and the activity of the enzymes FoF1-ATPase, 5′-nucleotidase, and AMP deaminase in the kidneys of rats exposed to different levels of protein and sucrose in their diet. Materials and Methods. The research was conducted on white non-linear rats, which were kept under different dietary regimens for a period of 4 weeks. Quantitative evaluation of the ATP, ADP, and AMP content was performed by thin-layer chromatography on Silufol sheets. FoF1-ATPase activity was determined by the accumulation of Pi. 5′-nucleotidase activity was measured based on the amount of inorganic phosphorus released in AMP hydrolysis. AMP deaminase activity was determined by the accumulation of ammonia. Results and Discussion. Research results revealed that in the mitochondria of the animals’ kidneys under conditions of low-protein diet, a significant reduction in AMP content was observed compared to the control, while ATP and ADP content remained unchanged. Simultaneously, the activities of 5′-nucleotidase, AMP deaminase, and FoF1-ATPase in the kidneys of animals on a low-protein diet were maintained at control levels. However, in rats maintained on a low-protein/high-sucrose diet, depletion of all adenine nucleotides is observed against an increase in the hydrolytic activity of FoF1-ATPase, AMP deaminase, and 5′-nucleotidase activities. The 5′-nucleotidase activity in animals of this group reaches maximum values in comparison with the control, indica­ting an enhanced AMP degradation mediated by 5′-nucleotidase in the conditions of low-protein/high-sucrose diet consumption. Conclusion. Excessive sucrose consumption in the context of dietary protein deficiency is accompanied by a depletion of the adenine nucleotides pool in the mitochondrial fraction and a significantly increased activity of purine catabolism enzymes in the cytosolic fraction of rat kidneys. This may result in an imbalance in the energy supply of renal cells. The obtained results open up prospects for developing a strategy for correc­ting energy metabolism disorders in the conditions of nutritional imbalance.

Hepato-splanchnic fluxes during exercise in patients with cirrhosis-a pilot study.

In cirrhotic patients, compromised hepatocyte function combined with disturbed hepatic blood flow could affect hepato-splanchnic substrate and metabolite fluxes and exacerbate fatigue during exercise. Eight cirrhotic patients performed incremental cycling trials (3 × 10 min; at light (28 [19-37] W; median with range), moderate (55 [41-69] W), and vigorous (76 [50-102] W) intensity). Heart rate increased from 68 (62-74) at rest to 95 (90-100), 114 (108-120), and 140 (134-146) beats/min (P < 0.05), respectively. The hepatic blood flow, as determined by constant infusion of indocyanine green with arterial and hepatic venous sampling, declined from 1.01 (0.75-1.27) to 0.69 (0.47-0.91) L/min (P < 0.05). Hepatic glucose output increased from 0.6 (0.5-0.7) to 1.5 (1.3-1.7) mmol/min, while arterial lactate increased from 0.8 (0.7-0.9) to 9.0 (8.1-9.9) mmol/L (P < 0.05) despite a rise in hepatic lactate uptake. Arterial ammonia increased in parallel to lactate from 47.3 (40.1-54.5) to 144.4 (120.5-168.3) μmol/L (P < 0.05), although hepatic ammonia uptake increased from 19.5 (12.4-26.6) to 69.5 (46.5-92.5) μmol/min (P < 0.05). Among the 14 amino acids measured, glutamate was released in the liver, while the uptake of free fatty acids decreased. During exercise at relatively low workloads, arterial lactate and ammonia levels were comparable to those seen in healthy subjects at higher workloads, while euglycemia was maintained due to sufficient hepatic glucose production. The accumulation of lactate and ammonia may contribute to exercise intolerance in patients with cirrhosis.

Fish production, water quality, and the role of nitrification as an ammonia removal process in intensively aerated hybrid catfish ponds

AbstractSix 0.1‐ha earthen ponds with 45 kW/ha of aeration capacity were stocked with hybrid catfish (Ictalurus punctatus x I. furcatus) ranging from 15,000 to 90,000 fish ha−1 in 15,000 fish ha−1 increments to analyze the effect of nitrogen loading on ammonia accumulation and fish production. Minimum dissolved oxygen (DO) concentration was maintained above 3.0 mg O2 L−1 to prevent hypoxia‐induced appetite reduction. Stocking density did not impact survival but decreased average fish harvest weight and increased food conversion ratio (FCR). Gross production increased with stocking density and ranged from 7249 to 35,674 kg ha−1. Average total (TAN) and un‐ionized ammonia (NH3) were not different among stocking densities despite greater nitrogen inputs in ponds with higher feeding rates. Nitrite () and nitrate () concentrations were significantly higher in ponds with higher stocking densities, suggesting that nitrification increased with nitrogen loading. Phytoplankton assimilation of ammonia is the primary ammonia removal process in traditional catfish ponds with modest fish densities and feeding rates. However, in intensively aerated ponds with high feeding rates, nitrification can become an important process for ammonia removal. These data indicate that small earthen hybrid catfish ponds with high aeration capacities can increase catfish production without ammonia accumulating.

Unraveling the role of urea hydrolysis in salt stress response during seed germination and seedling growth in Arabidopsis thaliana.

Urea is intensively utilized as a nitrogen fertilizer in agriculture, originating either from root uptake or from catabolism of arginine by arginase. Despite its extensive use, the underlying physiological mechanisms of urea, particularly its adverse effects on seed germination and seedling growth under salt stress, remain unclear. In this study, we demonstrate that salt stress induces excessive hydrolysis of arginine-derived urea, leading to an increase in cytoplasmic pH within seed radical cells, which, in turn, triggers salt-induced inhibition of seed germination (SISG) and hampers seedling growth. Our findings challenge the long-held belief that ammonium accumulation and toxicity are the primary causes of SISG, offering a novel perspective on the mechanism underlying these processes. This study provides significant insights into the physiological impact of urea hydrolysis under salt stress, contributing to a better understanding of SISG.

Phycosphere bacterial composition and function in colony and solitary Phaeocystis globosa strains providing novel insights into the algal blooms

Phycosphere bacteria can regulate the dynamics of different algal blooms that impact marine ecosystems. Phaeocystis globosa can alternate between solitary free-living cells and colonies and the latter morphotype is dominate during blooms. The mechanisms underlying the formation of these blooms have received much attention. High throughput sequencing results showed that the bacterial community composition differed significantly between colony and solitary strains in bacterial composition and function. It was found that the genera SM1A02 and Haliea were detected only among the colony strains and contribute to ammonium accumulation in colonies, and the genus Sulfitobacter was abundant among the colony strains that were excellent at producing DMS. In addition, the bacterial communities of the two colony strains exhibited stronger abilities for carbon and sulfur metabolism, energy metabolism, vitamin B synthesis, and signal transduction, providing inorganic and organic nutrients and facilitating tight communication with the host algae, thereby promoting growth and bloom development.

Precision control of ammonium release in Azotobacter vinelandii.

The capture and reduction of atmospheric dinitrogen gas to ammonium can be accomplished through the enzyme nitrogenase in a process known as biological nitrogen fixation (BNF), by a class of microbes known as diazotrophs. The diazotroph Azotobacter vinelandii is a model organism for the study of aerobic nitrogen fixation, and in recent years has been promoted as a potential producer of biofertilizers. Prior reports have demonstrated the potential to partially deregulate BNF in A. vinelandii, resulting in accumulation and extracellular release of ammonium. In many cases, deregulation requires the introduction of transgenic genes or elements to yield the desired phenotype, and the long-term stability of these strains has been reported to be somewhat problematic. In this work, we constructed two strains of A. vinelandii where regulation can be precisely controlled without the addition of any foreign genes or genetic markers. Regulation is maintained through native promoters found in A. vinelandii that can be induced through the addition of extraneous galactose. These strains result in varied degrees of regulation of BNF, and as a result, the release of extracellular ammonium is controlled in a precise, and galactose concentration-dependent manner. In addition, these strains yield high biomass levels, similar to the wild-type A. vinelandii strain and are further able to produce high percentages of the bioplastic polyhydroxybutyrate.

The effectiveness of garlic extracts on biogenic amine formation by foodborne pathogens and fish spoilage bacteria

SummaryImpacts of aqueous and ethanolic extracts of garlic were investigated in suppressing bacterial growth and biogenic amine (BA) formation by selected foodborne pathogens (Candida albicans, Salmonella paratyphi A, Escherichia coli and Staphylococcus aureus) and fish spoilage bacteria (Enterococcus faecalis, Photobacterium damselae and Pseudomonas luteola). The spread‐plate method was used to monitor bacterial growth in histidine decarboxylase broth (HDB), whereas the rapid high‐performance liquid chromatographic (HPLC) method was used for BA analysis. Bacterial growth and their ammonia and BA production were monitored using HDB. The results showed that bacterial growth on HDB was in the range from 9.13, for P. luteola, to 9.54 log CFU (colony‐forming units) mL−1, for S. aureus and C. albicans. The presence of garlic extracts in HDB resulted considerably in lowering bacterial growth and BA formation (P &lt; 0.05). The highest inhibitory activities of ethanolic and water garlic extracts were obtained for Gram‐positive S. aureus with 1.4 and 1.5 logarithmic reduction on bacterial growth, followed by Gram‐negative Salmonella Paratyphi A and E. coli. Application of garlic extracts, mainly ethanolic ones, showed a significant inhibitory effect on bacterial ammonia production, with 4‐100‐fold lower ammonia accumulation (P &lt; 0.05). Bacteria produced all tested BAs, mainly dopamine, agmatine and tryptamine. The highest levels of histamine and tyramine (61.99 and 36.45 mg L−1) were produced by S. aureus. In the presence of aqueous or ethanolic garlic extracts, putrescine production by E. faecalis was around 110‐fold lower than that of the control group. Results revealed that both garlic extracts are potent antimicrobials that can control the growth of foodborne pathogens and their harmful BA formation.

Cu(N2)-Li Battery for Ammonia Synthesis.

The cathodic mechanism of Li-N2 batteries is similar to Li-mediated N2 reduction (LiNR). Herein, the Li-N2, LiNR, and Cu-Li battery were amalgamated to a milliliter-scale Cu(N2)-Li system. The utilization of a lithium anode with lithium oxidation reaction (LiOR), ensures an uninterrupted supply of lithium ions to active N2. LiOR not only enhances electrolyte stability but also reduces voltage by stripping Li ions, in contrast to the inert platinum anode, commonly employed in LiNR. Notably, an unusual observation of ammonia accumulation within the anode chamber elucidates the presence and role of reaction intermediates. The charging process aimed at lithium regeneration faces high polarization, and a cycling procedure involving low-current charging was proposed to improve cycling. This study integrates insights from three distinct research directions to leverage their respective advantages and scientific insights. The Li-N2 battery emerges as a highly advantageous strategy for ammonia synthesis due to the progressiveness of lithium anode.

Linking microbial population dynamics in anaerobic bioreactors to food waste type and decomposition stage

A key question in anaerobic microbial ecology is how microbial communities develop over different stages of waste decomposition and whether these changes are specific to waste types. We destructively sampled over time 26 replicate bioreactors cultivated on fruit/vegetable waste (FVW) and meat waste (MW) based on pre-defined waste components and composition. To characterize community shifts, we examined 16S rRNA genes from both the leachate and solid fractions of the waste. Waste decomposition occurred faster in FVW than MW, as accumulation of ammonia in MW reactors led to inhibition of methanogenesis. We identified population succession during different stages of waste decomposition and linked specific populations to different waste types. Community analyses revealed underrepresentation of methanogens in the leachate fractions, emphasizing the importance of consistent and representative sampling when characterizing microbial communities in solid waste.

Impact of transport conditions and underlying disease on post-stocking survival of juvenile Lates calcarifer.

Diseases caused by pathogens commonly occurring in the aquatic environment or those that are non-host specific are prevalent and threaten the rapid growth of tropical aquaculture. This study investigates causes of mortality in 12 batches of newly stocked juvenile Lates calcarifer from three different hatcheries. Cytology based on Diff-Quik™-stained tissue and blood smears provides rapid diagnosis of possible causes of mortality, while histopathology and haematology provide a better understanding of how prolonged transport and fish with existing chronic disease are more likely to experience elevated mortality post-stocking. Our findings showed that accumulation of ammonia during prolonged transport causes extensive damage to epithelial barriers in gastrointestinal tracts and depressed immunity due to marked hypoglycaemia, predisposing fish to acute Streptococcosis. Lates calcarifer with chronic bacterial enteritis developed severe hypoglycaemia, had low circulating total plasma protein, and suffered high mortality within 24 hours post-stocking. Hypoglycaemia and low circulating blood proteins disrupt osmoregulation and exacerbate dehydration, which is fatal in fish in sea water. Dying L. calcarifer tested PCR positive for scale drop disease virus (SDDV) at 28 days post-stocking showed a 10-fold elevation of white blood cell counts, severe vasculitis, and obstruction of blood supply to major organs. Destruction of important immune organs such as spleen is a hallmark of SDDV infection that explains high incidences of opportunistic Vibrio harveyi infections in 61% of fish with SDDV. Overall, this study reiterates the importance of stocking disease-free fish and reducing transport stress.

Density‐dependent influence of ribbed mussels on salt marsh nitrogen pools and processes

Abstract Bivalves are becoming an increasingly popular tool to counteract eutrophication, particularly in vegetated coastal ecosystems where synergistic interactions between bivalves and plants can govern important N sequestration pathways. In turn, new calls to evaluate how bivalve densities modify N pools and processes across multiple scales have surfaced. Ribbed mussels, Geukensia demissa, and their relationship with smooth cordgrass present a classic demonstration of positive bivalve‐plant interactions and offer a useful model for assessing density dependence. We measure porewater ammonium concentrations, N stable isotope signatures in cordgrass tissue, and sediment N fluxes in mussel aggregations and in cordgrass‐only plots across a southeastern U.S. salt marsh. In addition to measuring the effect of mussel presence, we evaluate mussel density dependence through a multiscale approach. At the patch scale, we quantify mussel density effects within their aggregations (individuals m−2) while at a larger landscape scale, we quantify mussel density effects on the cordgrass‐only areas they neighbour (individuals ~30 m−2). Porewater ammonium concentrations were halved in mussel biodeposits relative to sediments in cordgrass‐only areas and negatively related to mussel density within aggregations. Leaf clip ẟ15N signatures were nearly 2‰ higher in cordgrass growing among mussel aggregations and increased with increasing patch mussel density. Microcosm incubations showed that mussels enhanced N2 flux (i.e., nitrogen removal) and DIN flux (i.e., N regeneration) into the water column, where only nitrogen removal increased with increasing patch‐scale mussel density. Across the marsh landscape, mussel coverage drove ammonium accumulation and N2 flux in sediments. Synthesis. Our results suggest that, at the patch scale, mussels stimulate the microbial metabolism of N, the assimilation of this bioavailable N by cordgrass, and nitrogen removal in a positive, density‐dependent manner. Tidal currents redistribute mussel biodeposits from mussel aggregations to surrounding areas, influencing biogeochemical transformations at scales beyond their physical footprint. We emphasize that the N regeneration potential of bivalve populations is a significant metric contributing to their mitigation potential and that bivalve density effects may be non‐linear, vary across patch to ecosystem scales, and have differing implications for the plants with which they interact.

Integrative omics analyses of tea (Camellia sinensis) under glufosinate stress reveal defense mechanisms: A trade-off with flavor loss

Extensively applied glufosinate (GLU) will trigger molecular alterations in nontarget tea plants (Camellia sinensis), which inadvertently disturbs metabolites and finally affects tea quality. The mechanistic response of tea plants to GLU remains unexplored. This study investigated GLU residue behavior, the impact on photosynthetic capacity, specialized metabolites, secondary pathways, and transcript levels in tea seedlings. Here, GLU mainly metabolized to MPP and accumulated more in mature leaves than in tender ones. GLU catastrophically affected photosynthesis, leading to leaf chlorosis, and decreased Fv/Fm and chlorophyll content. Physiological and biochemical, metabolomics, and transcriptomics analyses were integrated. Showing that GLU disrupted the photosynthetic electron transport chain, triggered ROS and antioxidant system, and inhibited photosynthetic carbon fixation. GLU targeted glutamine synthetase (GS) leading to the accumulation of ammonium and the inhibition of key umami L-theanine, causing a disorder in nitrogen metabolism, especially for amino acids synthesis. Interestingly, biosynthesis of primary flavonoids was sacrificed for defensive phenolic acids and lignin formulation, leading to possible losses in nutrition and tenderness in leaves. This study revealed the defense intricacies and potential quality deterioration of tea plants responding to GLU stress. Valuable insights into detoxification mechanisms for non-target crops post-GLU exposure were offered.

Ammonia Accumulation as a Proxy to Determine Cage-change Frequency in Antelope Ground Squirrels (Ammospermophilus leucurus).

Due to a lack of evidence-based standards for cage-change intervals for antelope ground squirrels (AGS, Ammospermophilus leucurus), we evaluated cage ammonia accumulation in our colony of adult, wild-caught AGS and identified factors that influenced ammonia levels. Intracage ammonia was measured daily in singly housed AGS in static caging that contained a running wheel and 1/2, 3/4, 1, or 2 quart (qt) of corncob bedding. Cages were changed when ammonia levels reached greater than 50ppm, our upper acceptable limit for ammonia based on mouse studies of ammonia aversion and toxicity. We also measured average daily water consumption over 2 wk to examine any correlation between water use and ammonia accumulation. We hypothesized that the desert-dwelling AGS would not reach intracage ammonia levels of greater than 50ppm in a 2-wk interval at any bedding volume. Our data showed that intracage ammonia was highly variable among individuals and was significantly associated with water consumption and bedding volumes. Seventeen percent of AGS on 1/2qt of bedding and 18% on 3/4qt of bedding reached greater than 50ppm ammonia before 7 d. All AGS on 1 and 2qt of bedding remained below 50ppm ammonia for 1 wk. Even when maintained on 2qt of bedding, not all AGS remained below 50ppm ammonia for 2 wk. Therefore, we concluded that the most appropriate option was weekly cage change for singly housed AGS on 1qt of bedding in static caging.

Ammonia-dependent reducing power redistribution for purple phototrophic bacteria culture-based biohydrogen production

The global energy crisis has intensified the search for sustainable and clean alternatives, with biohydrogen emerging as a promising solution to address environmental challenges. Leveraging photo fermentation (PF) process, purple phototrophic bacteria (PPB) can harness reducing power derived from organic substrates to facilitate hydrogen production. However, existing studies report much lower H2 yields than theoretical value when using acetate as carbon source and ammonia as nitrogen source, primarily attributed to the widely employed pulse-feeding mode which suffers from ammonia inhibition effect on nitrogenase. To address this issue, a continuous feeding mode was applied to avoid ammonia accumulation in this study. On the other hand, other pathways like carbon fixation and polyhydroxyalkanoate (PHA) formation could compete reducing power with H2 production. However, the reducing power allocation under continuous feeding mode is not yet clear. In this study, the reducing power allocation and hydrogen production performance were evaluated under various ammonia loading, using acetate as carbon source and infrared LED at around 50 W·m−2 as light source. The results show that (a) The absence of ammonia resulted in the best performance for hydrogen production, with 44 % of the reducing power distributed to H2 and the highest H2 volumetric productivity, while the allocation of reducing power to hydrogen production stopped when ammonia loading was above 7.6 mg NH4-N·L−1·d−1; (b) when PPB required to eliminate reducing power under ammonia limited conditions, PHA production was the preferred pathway followed by the hydrogen production pathway, but once PHA accumulation reached saturation, hydrogen generation pathway dominated; (c) under ammonia limited conditions, the TCA cycle was more activated rendering higher NADH (i.e. reducing power) production compared with that under ammonia sufficient conditions which was verified by metagenomics analysis, and all the hydrogen production, PHA accumulation and carbon fixation pathways were highly active to dissipate reducing power. This work provides the insight of reducing power distribution and PPB biohydrogen production variated by ammonia loading under continuous feeding mode.