Acid-Base Effects of Combined Renal Deletion of NBCe1-A and NBCe1-B

The molecular mechanisms regulating ammonia metabolism are fundamental to acid-base homeostasis. Deleting the A splice variant of the Na⁺-bicarbonate cotransporter, electrogenic, isoform 1 (NBCe1-A) partially blocks the effect of acidosis to increase urinary ammonia excretion, and this appears to involve the dysregulated expression of ammoniagenic enzymes in the proximal tubule (PT) in the cortex, but not in the outer medulla (OM). A second NBCe1 splice variant, NBCe1-B, is present throughout the PT, including the OM, where NBCe1-A is not present. The current studies determined the effects of combined renal deletion of NBCe1-A and NBCe1-B on systemic and proximal tubule ammonia metabolism. We generated NBCe1-A/B deletion using Cre-loxP techniques and used Cre-negative mice as controls. Since renal NBCe1-A and NBCe1-B expression is limited to the proximal tubule, Cre-positive mice had proximal tubule NBCe1-A/B deletion (PT-NBCe1-A/B KO). While on basal diet, PT-NBCe1-A/B KO mice had severe metabolic acidosis, yet urinary ammonia excretion was not changed significantly. PT-NBCe1-A/B KO decreased expression of phosphate-dependent glutaminase (PDG) and phospho­enol­pyruvate carboxy­kinase (PEPCK) and increased expression of glutamine synthetase (GS), an ammonia recycling enzyme, in PT in both the cortex and OM. Exogenous acid-loading increased ammonia excretion in control mice, but PT-NBCe1-A/B KO prevented any increase. PT-NBCe1-A/B KO significantly blunted acid loading-induced changes in PDG, PEPCK, and GS expression in the proximal tubule in both the cortex and OM. We conclude that NBCe1-B, at least in the presence of NBCe1-A deletion, contributes to proximal tubule ammonia metabolism in the OM and thereby to systemic acid-base regulation.

Influence of diet and feeding strategy on the performance of nitrifying trickling filter, oxygen consumption and ammonia excretion of gilthead sea bream (Sparus aurata) raised in recirculating aquaculture systems

Gilthead sea bream (Sparus aurata) was raised in six individual recirculating aquaculture systems (RAS) whose biofilters’ performance was analyzed. Fish were fed with three different diets (a control diet, a fishmeal-based diet (FM), and a plant meal-based diet (VM)) and with three different feeding strategies (manual feeding to apparent satiation, automatic feeding with restricted ration, and auto-demand feeding). For every combination of diet and feeding strategy, the mean oxygen consumption, ammonia excretion, and ammonia removal rate were determined. Fish fed with the VM diet consumed the most oxygen (20.06 ± 1.80 gO2 consumed kg−1 day−1). There were significant differences in ammonia excretion depending on the protein content and protein efficiency of the diet, as well as depending on feeding strategy, which in turn affected ammonia removal rates. Fish fed by auto-demand feeders led to the highest mean ammonia removal rate (0.10 gN-TAN removed m−2 biofiltration area day−1), while not leading to peaks of high ammonia concentration in water, which preserve fish welfare and growth.

Isolation and Characterization of Effective Bacteria That Reduce Ammonia Emission from Livestock Manure

Ammonia from livestock manure reacts with chemical components discharged from various emission sources to produce airborne particulate matter. This study aimed to investigate a novel effective microbial agent to suppress ammonia gas emitted from manure. Both isolated L12I and 12III strains, identified as Pediococcus acidilactici (PA), were selected for their superior activity in assays performed with the evaluation criteria such as acid production, ammonia decomposition, and urease inhibition, which are key factors influencing ammonia excretion. The survivability of PA strains was confirmed by an increase in DNA abundance in the manure. PA strains lowered the pH of manure and suppressed the growth of hyper-ammonia-producing bacteria (HAB) possessing urease activity. The L12I and 12III treatment groups showed 23.58% and 38.00% emission reductions, respectively. Especially, the 12III strain was proven to be the more effective strain for reducing ammonia gas emission, with the best ability to reduce pH and inhibit HAB. The strains could have an additive effect in improving the manure quality as a nitrogen fertilizer by preserving the total nitrogen and urea content. These results suggest that PA strains can be used as unprecedented microbial agents to improve manure-derived environmental pollution and improve fertilizer quality.

Physiological and Molecular Responses in the Gill of the Swimming Crab Portunus trituberculatus During Long-Term Ammonia Stress

Ammonia is a common environmental stressor encountered during aquaculture, and is a significant concern due to its adverse biological effects on vertebrate and invertebrate including crustaceans. However, little information is available on physiological and molecular responses in crustaceans under long-term ammonia exposure, which often occurs in aquaculture practices. Here, we investigated temporal physiological and molecular responses in the gills, the main ammonia excretion organ, of the swimming crab Portunus trituberculatus following long-term (4 weeks) exposure to three different ammonia nitrogen concentrations (2, 4, and 8 mg l–1), in comparison to seawater (ammonia nitrogen below 0.03 mg l–1). The results revealed that after ammonia stress, the ammonia excretion and detoxification pathways were initially up-regulated. These processes appear compromised as the exposure duration extended, leading to accumulation of hemolymph ammonia, which coincided with the reduction of adenosine 5′-triphosphate (ATP) and adenylate energy charge (AEC). Considering that ammonia excretion and detoxification are highly energy-consuming, the depression of these pathways are, at least partly, associated with disruption of energy homeostasis in gills after prolonged ammonia exposure. Furthermore, our results indicated that long-term ammonia exposure can impair the antioxidant defense and result in increased lipid peroxidation, as well as induce endoplasmic reticulum stress, which in turn lead to apoptosis through p53-bax pathway in gills of the swimming crab. The findings of the present study further our understanding of adverse effects and underlying mechanisms of long-term ammonia in decapods, and provide valuable information for aquaculture management of P. trituberculatus.

Comparison in Growth, Feeding, and Metabolism Between a Fast-Growing Selective Strain and a Cultured Population of Pearl Oyster (Pinctada fucata martensii)

Pearl oyster (Pinctada fucata martensii) is the main species cultured for marine pearls in the world. A breeding program was carried out for desirable production traits, including high growth rate, and a fast-growing selective strain of pearl oysters was established. In the current study, we compared the growth characteristics between a selective strain and a cultured population of P. f. martensii in Beihai, Guangxi Province, China. Large size (SL) and small size (SS) individuals of the selective strain were selected, and the differences of physiological and metabolic indexes, such as feeding, respiration, excretion, and enzyme activities between SL and SS and cultured population (CL), were also compared. The results showed that at the age of 6 months, pearl oysters of the selective strain were 14.61% larger than CL, and the proportion of SL (30–40 mm) was 59%, which was two times higher than CL (28%). SL with a rapid growth rate had a high clearance rate (CR), and the CR of SL was about 1.8 times higher than that of CL and 5 times higher than that of SS. In addition, the activities of digestive enzymes (amylase, pepsin, and lipase) and growth-related carbonic anhydrase enzymes in SL were higher than those in the other two groups (p < 0.05). SS with a slow growth rate had higher oxygen consumption (OCR) and ammonia excretion (AER) rates than SL and CL (p < 0.05). Our results suggest that the rapid growth of the selective strain P. f. martensii can be attributed to increased energy intake and reduced energy consumption.

Study the effect of low pH on Ruditapes decussatus (Mollusca: Bivalvia)

The grooved carpet shell clam (Ruditapes decussatus) is one of the most economically essential mollusks in Mediterranean lagoons and sandy beaches, with fisheries and aquaculture both contributing to its abundance. The goal of this research is to see how varying amounts of acidity affect this calcifying organism. 420 ppm (ambient control), 550 ppm, 750 ppm, and 1050 ppm were used to incubate juvenile clams in CO2 enriched saltwater. With increasing pCO2 , the biological parameters evaluated revealed a small decline. Differences, however, were not substantial. In terms of overall weight, the reduction was greatest at 550 and 1050 ppm. Furthermore, clams kept at 550 parts per million had the lowest condition index and the greatest mortality rate of 8%. Both the 550 ppm and the control 420 ppm groups demonstrated an increase in metabolic rate and ammonia excretion in the physiological response testing. With increasing acidification, the algal feed clearance rate declined, with the highest average value in the control (420 ppm) group and the lowest average value in the extremely high pCO2 (1050 ppm) group. Ocean acidification may put further strain on R. decussatus' health and economic value by the end of the century

Ascaris suum Nutrient Uptake and Metabolic Release, and Modulation of Host Intestinal Nutrient Transport by Excretory-Secretory and Cuticle Antigens In Vitro

Ascaris suum, the most important pig parasite, also infects humans as a zoonotic pathogen. Malabsorption upon infection probably results from impaired nutrient transport, presumably mediated by the parasite´s excretory-secretory (ES) or cuticle somatic (CSO) antigens. The present study investigated the electrogenic transport (∆Isc) of glucose, alanine and the dipeptide glycyl-l-glutamine (glygln), as well as glucose net flux rates in pig jejunal tissue after in vitro exposure to adult A. suum total ES or CSO antigens in Ussing chambers. ∆Isc of glucose, alanine and glucose net flux rate were significantly decreased after one hour of exposure to total ES antigen. In contrast, CSO antigens increased the transport of glygln. Additionally, nutrient uptake and ES antigen pattern were compared in culture medium from untreated adult worms and those with sealed mouth and anal openings. Untreated worms completely absorbed glucose, while cuticular absorption in sealed worms led to 90% reduction. Amino acid absorption was 30% less effective in sealed worms, and ammonia excretion decreased by 20%. Overall, the results show that A. suum total ES antigen rapidly impairs nutrient transport in vitro. Future studies confirming the results in vivo, narrowing down the ES components responsible and investigating underlying molecular mechanisms are needed.

Oxygen Consumption and Ammonia Excretion of Marphysa sanguinea (Polychaeta: Eunicidae) in Relation to Body Mass and Temperature

(1) Background: Marphysa sanguinea is a polychaete with high economic value and ecological importance. Information on metabolism is important to understand the physiological action of organisms. (2) Methods: The rates of oxygen consumption (R) and ammonia excretion (U) were measured using different temperatures (T) and body mass (M) levels. The activation energy (E) was calculated using the universal temperature dependence theory. (3) Results: Oxygen consumption presented a curve with an upward trend first, and then a downward trend, and ammonia excretion displayed a “U” curve. The effects of temperature and body size on oxygen consumption and ammonia excretion rates were extremely significant. Small individuals had higher metabolic rates than large polychaetes at the same temperature. The relationship between oxygen consumption, ammonia excretion, and M was expressed as Y = a·Mb, bR = 0.56 ± 0.09, and bU = 0.35 ± 0.30. The oxygen consumption activation energy was ER = 0.68 eV, and the ammonia excretion activation energy was EU = 0.53 eV. The O:N ratio at different temperatures and body sizes was in the range of 3.55–56.44. (4) Conclusions: The results not only provide basic data on the metabolism of M. sanguinea but also insights to understand the relationship between animal metabolism and ecological factors from different perspectives.

Role of the Renal Androgen Receptor in Ammonia Metabolism

Background: There are sex differences in renal ammonia metabolism and structure, many of which are mediated by testosterone. This study's goal was to determine the role of renal expression of testosterone's canonical receptor, androgen receptor (AR), in these sexual dimorphisms. Methods: We studied mice with kidney-specific AR deletion (KS-AR-KO) generated using Cre/loxP techniques; control mice were Cre-negative littermates (WT). Results: In male, but not female, mice, KS-AR-KO increased ammonia excretion, which eliminated sex differences. Although renal structural size typically parallel ammonia excretion, KS-AR-KO decreased kidney size, cortical proximal tubule volume density and cortical proximal tubule cell height in males; neither were altered in females and collecting duct volume density was unaltered in both sexes. Analysis of key protein involved in ammonia handling showed in male mice that KS-AR-KO increased both PEPCK and NKCC2 expression, and decreased NHE3 and NBCe1-A expression. In female mice, KS-AR-KO did not alter these parameters. These effects occurred even though KS-AR-KO did not alter plasma testosterone, food intake or serum Na+, K+, or HCO3- significantly in either sex. Conclusions: AR-dependent signaling pathways in male, but not female, kidney regulate PEPCK and NKCC2 expression and lead to the sexual differences in ammonia excretion. Opposing effects on NHE-3 and NBCe1-A expression likely limit the magnitude of ammonia excretion changes. Since AR is not present in the TAL, the effect of KS-AR-KO on NKCC2 expression is indirect. Finally, AR mediates the greater kidney size and PT volume density in male than in female mice.

Role of the cardiovascular system in ammonia excretion in early life stages of zebrafish (Danio rerio)

The purpose of this study was to investigate if the cardiovascular system is important for ammonia excretion in the early life stages of zebrafish. Morpholino knockdowns of cardiac troponin T (TNNT2) or vascular endothelial growth factor (VEGFA) provided morphants with non-functional circulation. At the embryonic stage (30-36 hpf), ammonia excretion was not constrained by lack of cardiovascular function. At 2 days post fertilization (dpf) and 4 dpf, morpholino knockdowns of TNNT2 or VEGFA significantly reduced ammonia excretion in all morphants. Expression of rhag, rhbg and rhcgb showed no significant changes but the mRNA levels of the urea transporter (ut) were upregulated in the 4 dpf morphants. Taken together, rhag, rhbg, rhcgb and ut gene expression and an unchanged tissue ammonia concentration but an increased tissue urea concentration, suggest that impaired ammonia excretion led to increased urea synthesis. However, in larvae anesthetized with tricaine or clove oil, ammonia excretion was not reduced in the 4 dpf morphants compared to controls. Further, oxygen consumption was reduced in morphants regardless of anesthesia. These results suggest that cardiovascular function is not directly involved in ammonia excretion, but rather reduced activity and external convection may explain reduced ammonia excretion and compensatory urea accumulation in morphants with reduced cardiovascular function.