Combined effects of hypoxia and ammonia-N exposure on the oxygen consumption, glucose metabolism and amino acid metabolism in hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂).

This study evaluated the influence of hypoxia and ammonia-N co-exposure on oxygen consumption, glucose metabolism and amino acid metabolism in hybrid grouper. The results showed that elevated expression of GLUT1, MCT1, PFK, HK and LDH were induced by co-exposure to hypoxia and ammonia. In addition, co-exposure to hypoxia and ammonia reduced the tolerance of hybrid grouper to ammonia-N. Furthermore, ammonia-N exposure caused an increase in oxygen consumption in hybrid grouper. After ammonia-N exposure for 96 h, 10 amino acids contents and activities of AST and ALT elevated in hybrid grouper muscle. The study revealed that combined exposure to hypoxia and ammonia-N significantly increased glucose metabolism, oxygen consumption and amino acid metabolism in hybrid grouper, and presented significant synergistic effects.

Evaluation of Genetic Parameters and Comparison of Stress Tolerance Traits in Different Strains of Litopenaeus vannamei.

Litopenaeus vannamei stands out globally in aquaculture for its fast growth, broad salt tolerance, disease resistance, and high protein levels. Selective breeding requires the precise estimation of the variance components and genetic parameters for important traits. This study formed lineages from 20 full sibling families of L. vannamei, with progenitors from Thailand and the USA. We then assessed the genetic resilience traits of juvenile shrimp from these families to high ammonia-N, high pH, and low salinity by performing a 96 h acute toxicity test. Mortality rates for the families under 96 h exposure to high ammonia-N, high pH, and low salinity were 19.52-92.22%, 23.33-92.22%, and 19.33-80.00%, respectively, showing significant variance in stress tolerance among families (p < 0.05). Survival heritability estimates, using threshold male and female models, were 0.44 ± 0.12 in high ammonia-N, 0.41 ± 0.12 in high pH, and 0.27 ± 0.08 in low salinity, respectively. Genetic correlations between growth and stress resistance traits varied from 0.0137 ± 0.2406 to 0.8327 ± 0.0781, and phenotypic correlations ranged from 0.0019 ± 0.0590 to 0.6959 ± 0.0107, indicating a low-to-high positive correlation significant at (p < 0.05). It was found that the survival rate of families No. 2 and No. 9 was higher under high ammonia-N and high pH stresses, while the survival rate of family No. 10 was higher under low salinity stress after comparing two selection criteria, the breeding values and phenotypic values. Thus, these three families are identified as potential breeding program candidates. Through the creation of a genetic parameter estimation model, the genetic variances across mating combinations for stress resistance traits were obtained and families with heightened stress resistance were identified, laying the groundwork for enhanced genetic selection of L. vannamei.

Mechanisms of ammonotelism, epithelium damage, cellular apoptosis, and proliferation in gill of Litopenaeus vannamei under NH4Cl exposure.

Excessive ammonia-N in coastal environment and aquaculture threatens the health of marine organisms. To explore the mechanism of gill damage induced by ammonia-N, transcriptome of Litopenaeus vannamei 's gill was carried out under 20 mg/L NH4Cl for 0, 6, and 48 h. K-means clustering analysis suggested that ammonia excretion and metabolism-related genes were elevated. GO and KEGG enrichment analysis suggested that glycosyltransferase activity and amino acid metabolism were affected by ammonia. Moreover, histological observation via three staining methods gave clues on the changes of gill after ammonia-N exposure. Increased mucus, hemocyte infiltration, and lifting of the lamellar epithelium suggested that gill epithelium was suffering damage under ammonia-N stress. Meanwhile, the composition of extracellular matrix (ECM) in connective tissue changed. Based on the findings of transcriptomic and histological analysis, we further investigated the molecular mechanism of gill damage under multiple concentrations of NH4Cl (0, 2, 10, 20 mg/L) for multiple timepoints (0, 3, 6, 12, 24, 48, 72 h). First, ammonia excretion was elevated via ion channel, transporter, and exocytosis pathways, but hemolymph ammonia still kept at a high level under 20 mg/L NH4Cl exposure. Second, we focused on glycosaminoglycan metabolism which was related to the dynamics of ECM. It turned out that the degradation and biosynthesis of chondroitin sulfate (CS) were elevated, suggesting that the structure of CS might be destructed under ammonia-N stress and CS played an important role in maintaining gill structure. It was enlightening that the destructions occurred in extracellular regions were vital to gill damage. Third, ammonia-N stress induced a series of cellular responses including enhanced apoptosis, active inflammation, and inhibited proliferation which were closely linked and jointly led to the impairment of gill. Our results provided some insights into the physiological changes induced by ammonia-N and enriched the understandings of gill damage under environmental stress.

Differential Toxicity Responses between Hepatopancreas and Gills in Litopenaeus vannamei under Chronic Ammonia-N Exposure.

Ammonia nitrogen is one of the main toxic substances in aquatic cultivation environments. Chronic exposure to excessive amounts of ammonia-N creates toxic consequences, retarding the growth of aquatic organisms. This study investigated the growth performance, morphological and physiological alterations, and transcriptome changes in the hepatopancreas and gills of white shrimp Litopenaeus vannamei. The results showed that there was no significant difference in the survival rate (p > 0.05), whereas growth performance was reduced significantly in the treated groups compared to the control groups (p < 0.05). Significant structural damage and vacuolation occurred in hepatopancreas and gill tissues in the treated groups. Superoxide dismutase (SOD) activity and Na+/K+-ATPase content were significantly increased by chronic ammonia-N exposure in the two tissue groups. In addition, catalase (CAT) activity and malondialdehyde (MDA) levels were significantly altered in the hepatopancreas groups (p < 0.05), whereas no differences were observed in the gill groups (p > 0.05). There were 890 and 1572 differentially expressed genes identified in the hepatopancreas (treated versus control groups) and gills (treated versus control groups), respectively, of L. vannamei under chronic ammonia-N exposure. Functional enrichment analysis revealed associations with oxidative stress, protein synthesis, lipid metabolism, and different serine proteases. The gills maintained cellular homeostasis mainly through high expression of cytoskeleton and transcription genes, whereas the hepatopancreas down-regulated related genes in the ribosome, proteasome, and spliceosome pathways. These genes and pathways are important in the biosynthesis and transformation of living organisms. In addition, both tissues maintained organismal growth primarily through lipid metabolism, which may serve as an effective strategy for ammonia-N resistance in L. vannamei. These results provided a new perspective in understanding the mechanisms of ammonia-N resistance in crustaceans.

In vitro protein fractionation methods for ruminant feeds.

Estimating protein fractions and their degradation rate are vital to ensure optimum protein supply and degradation in the digestive system of ruminants. This study investigated the possibility of using the ANKOM gas production system and preserved rumen fluid to estimate the protein fractions and in vitro degradability of protein-rich feeds. Three in vitro methods: (1) gas production method (2) Cornell Net Carbohydrate and Protein System (CNCPS), and (3) the unavailable nitrogen assay of Ross (uNRoss) were used to quantify protein fractions of four feeds (lupin meal, vetch grain, Desmanthus hay, and soybean meal). Rumen fluid mixed with 5% dimethyl sulfoxide and frozen at -20 °C was also compared against fresh rumen fluid in the gas production and uNRoss methods. All three methods ranked the feeds identically in the proportions of available (degradable or 'a + b') protein fractions as vetch grain, soybean meal, lupin meal, and Desmanthus hay in decreasing order. The use of fresh rumen fluid produced greater available protein fractions than preserved rumen fluid in all feeds. However, there was no difference between total gas production from lupin meal and vetch grain fermented for 16 h in either rumen fluid source. The in vitro degradable CP (IVDP) was higher for vetch grain (46 and 70%) at the 4th and 8th hours of incubation than other feeds, whereas soybean meal (85%) exceeded the other feeds after the 16th hour of incubation (P < 0.001). The greatest ammonia-N concentration was from soybean meal (1.27 mg/g) and lupin meal (0.87 mg/g) fermented for four hours using fresh rumen fluid. The proportion of fraction 'b' for soybean (82.1% CP) and lupin meals (39.4% CP) from the CNCPS method were not different (P = 0.001) from the fraction 'b' estimation of the gas production method for the same feeds (r = 0.99). Regardless of the methods, a greater water-soluble protein fraction was found from vetch grain (39.6-46.6% CP), and the proportion of fraction 'c' or unavailable protein in Desmanthus hay (39.1-41.5% CP) exceeded other substrates (P < 0.001). The strong positive correlation between fractions across different methods and identical ranking of feeds suggests the possibility of using ANKOM gas production apparatus for protein fractionation.

First transcriptome profiling in gill and hepatopancrease tissues of Metapenaeus ensis in response to acute ammonia-N stress.

The greasyback shrimp, Metapenaeus ensis, suffers from ammonia-N stress during intensive factory aquaculture. Optimizing ammonia-N stress tolerance has become an important issue in M. ensis breeding. The metabolic and adaptive mechanisms of ammonia-N toxicity in M. ensis have not been comprehensively understood yet. In this study, a large number of potential simple sequence repeats (SSRs) in the transcriptome of M. ensis were identified. Differentially expressed genes (DEGs) in the gill and hepatopancreas at 24 h post-challenges under high concentrations of ammonia-N treatment were detected. We obtained 20,108,851-27,681,918 clean reads from the control and high groups, assembled and clustered a total of 103,174 unigenes with an average of 876 bp and an N50 of 1189 bp. Comparative transcriptome analyses identified 2000 different expressed genes in the gill and 2010 different expressed genes in the hepatopancreas, a large number of which were related to immune function, oxidative stress, metabolic regulation, and apoptosis. The results suggest that M. ensis may counteract ammonia-N toxicity at the transcriptome level by increasing the expression of genes related to immune stress and detoxification metabolism, and that selected genes may serve as molecular indicators of ammonia-N. By exploring the genetic basis of M. ensis' ammonia-N stress adaptation, we constructed the genetic networks for ammonia-N adaptation. These findings will accelerate the understanding of M. ensis' ammonia-N adaptation, contribute to the research of future breeding, and promote the level of factory aquaculture of M. ensis.

Exercise and emersion in air and recovery in seawater in the green crab (Carcinus maenas): Effects on nitrogenous wastes and branchial chamber fluid chemistry.

At low tide, the green crab, which is capable of breathing air, may leave the water and walk on the foreshore, carrying branchial chamber fluid (BCF). N-waste metabolism was examined in crabs at rest in seawater (32 ppt, 13°C), and during 18-h recovery in seawater after 1 h of exhaustive exercise (0.25 BL s-1 ) on a treadmill in air (20°C-23°C), or 1 h of quiet emersion in air. Measurements were made in parallel to O2 consumption (MO2 ), acid-base, cardio-respiratory, and ion data reported previously. At rest, the ammonia-N excretion rate (MAmm = 44 µmol-N kg-1 h-1 ) and ammonia quotient (AQ; MAmm /MO2 = 0.088) were low for a carnivore. Immediately after exercise and return to seawater, MAmm increased by 65-fold above control rates. After emersion alone and return to seawater, MAmm increased by 17-fold. These ammonia-N bursts were greater, but transient relative to longer-lasting elevations in MO2 , resulting in temporal disturbances of AQ. Intermittent excretion of urea-N and urate-N at rest and during recovery indicated the metabolic importance of these N-wastes. Hemolymph glutamate, glutamine, and PNH3 did not change. Hemolymph ammonia-N, urea-N, and urate-N concentrations increased after exercise and more moderately after emersion, with urate-N exhibiting the largest absolute increments, and urea-N the longest-lasting elevations. All three N-wastes were present in the BCF, with ammonia-N and PNH3 far above hemolymph levels even at rest. BCF volume declined by 34% postemersion and 77% postexercise, with little change in osmolality but large increases in ammonia-N concentrations. Neither rapid flushing of stored BCF nor clearance of hemolymph ammonia-N could explain the surges in MAmm after return to seawater.

Nuclear factor interleukin 3 (NFIL3) participates in regulation of the NF-κB-mediated inflammation and antioxidant system in Litopenaeus vannamei under ammonia-N stress.

Nuclear factor interleukin 3 (NFIL3) is a critical upstream regulator of the NF-κB pathway. Nevertheless, the detailed molecular mechanism of NFIL3 and its function in shrimp have not been well characterized. In the present study, NFIL3 was identified and characterized from Litopenaeus vannamei. Molecular feature analysis revealed that the open reading frame (ORF) of LvNFIL3 was 2963 bp, which codes for a polypeptide of 516 amino acids with a conserved basic region leucine zipper (bZIP) domain. Sequence alignments and phylogenetic tree analysis showed that the amino acid sequence of LvNFIL3 shared 18.82%-98.07% identity with that of NFIL3 in other species, and was closely related to Penaeus monodon NFIL3. A core promoter in the 5' flanking region of LvNFIL3 was essential for regulation of transcription. LvNFIL3 mRNA was highly expressed in gills and hepatopancreas. Subcellular localization of the protein was observed almost exclusively in the nucleus. Amplification of mRNA by RT-qPCR showed that LvNFIL3 was induced in shrimp gills, hepatopancreas, and muscle after ammonia-N stress. Moreover, silencing of LvNFIL3 increased the mortality of shrimp exposed to ammonia-N. Furthermore, dual-luciferase reporter assay data suggested that LvNFIL3 was capable of activating the NF-κB pathway. Conversely, knockdown of LvNFIL3 decreased NF-κB homolog (Dorsal and Relish) and IkB homolog (Cactus) expression, as well as expression of anti-inflammatory cytokine (IL-16) and five antioxidant-related genes (HO-1, Mn-SOD, CAT, GPx, and GST), whereas NF-κB repressing factor (NKRF) and inflammation-related genes (TNFα and Spz) were upregulated. More importantly, LvNFIL3 knockdown exacerbated the pathology in hepatopancreas exposed to ammonia-N, and the total antioxidant capacity (T-AOC) and superoxide dismutase (T-SOD) were significantly decreased, resulting in a significant increased lipid peroxidation and protein carbonization. Taken together, these data suggest that LvNFIL3 was involved in ammonia-N tolerance in L. vannamei by regulating the inflammation and antioxidant system through the NF-κB pathway.

Field pea can replace soybean meal-corn mixtures in the fattening concentrate of young bulls improving the digestibility.

The inclusion of pea (Pisum sativum) in the fattening concentrate at 0%, 15%, 30%, and 45% as a replacement for soybean meal and corn was evaluated on in vitro and in vivo digestibility studies of young Parda de Montaña bulls. In the in vitro trial, gas production was determined with an Ankom system for 48 h. The 30%pea and 45%pea concentrates increased the organic matter (OM) degradability, the ammonia-N content and the ratio of acetic:propionic, with no effect on gas production, final ruminal pH and total volatile fatty acid production. In the in vivo assay, 4 young bulls (initial weight 251 ± 4 kg) received restricted amounts of concentrates plus straw during 4 consecutive experimental periods. No differences in intake were observed and 30%pea and 45%pea had higher digestibility of crude protein (CP), and OM than the rest of treatments. The nitrogen intake increased linearly with the inclusion of pea with similar nitrogen urinary and faecal excretions, and the nitrogen retained was greater in 30%pea and 45%pea than the rest of treatments. The plasma concentrations of IGF-1, total protein, ß-hydroxybutyrate and urea at the beginning and at the end of each period were not affected by the inclusion of pea. In conclusion, the total replacement of soybean-corn mixtures by pea in the fattening concentrate of bulls could be recommended as it improved the CP digestibility and nitrogen retention.

Application of response surface methodology for COD and ammonia removal from municipal wastewater treatment plant using acclimatized mixed culture.

This study aimed to optimize conditions influencing the removal of chemical oxygen demand (COD) and ammonia-N in municipal wastewater by using acclimatized mixed culture (AMC). Two-level factorial analysis was used to investigate the factors affecting the degradation of COD and ammonia-N (%); ratio of synthetic wastewater (SW) to acclimatized mixed culture (AMC) (1:1 and 3:1), presence and absence of support media (Yes and No), agitation (0 rpm and 100 rpm) and hydraulic retention time (HRT) (2 and 5 days). A central composite design (CCD) under response surface methodology (RSM) determined the optimum agitation (0 rpm and 100 rpm) and retention time (2 and 5 days). The best conditions were at 3:1 of SW: AMC ratio, 100 rpm agitation, without support media, and 5 days retention time. COD and ammonia-N removal achieved until 57.23% and 43.20%, respectively. Optimization study showed the optimum conditions for COD and ammonia-N removal were obtained at 150 rpm agitation speed and 5 days of retention time, at 70.41% and 64.29% respectively. This study discovers the conditions that affect the COD and ammonia-N removal in the municipal wastewater using acclimatized mixed culture.

Effects of ammonia-N stress on molecular mechanisms associated with immune behavior changes in the haemocytes of Litopenaeus vannamei.

High concentration of ammonia-N will inhibit the immune defense of aquatic animals. The neuroendocrine-immune (NEI) regulatory mechanism under ammonia-N stress has been systematically studied, but the final response mechanism of ammonia-N affecting the immune system remains unclear. To investigate the relationship among immune factors of Litopenaeus vannamei (L. vannamei) exposed to 0, 2, 10 and 20 mg/L ammonia-N, the determination of complement components, C-type lectins, proPO system, signal transduction pathway and phagocytosis as well as exocytosis were performed. The results showed that the expressions of complement components including C1q, MBL, ficolin and alpha-2 macroglobulin (A2M) and the complement receptor integrin were decreased significantly in ammonia-N treatment groups at 6,12 and 24 h. C-type lectins and signal transduction factors changed significantly. The decrease of phagocytosis-related genes and phagocytic activity were similar to the changes of complement components, C-type lectins and the signal pathway. The mRNA abundance of exocytosis-related genes was significantly down-regulated under ammonia-N exposure. Correspondingly, significantly changes occurred in the expressions of PPAE and PPO3, immune factors-related genes (Pen3, crustin, stylicins, ALFs and LYC) and inflammatory factors (HSP90, TNFα, IL-16) in haemocytes. Eventually, the serine proteinase activity, PO activity, antibacterial activity and bacteriolytic activity in plasma were decreased significantly. In addition, we speculated that under ammonia-N stress, phagocytosis and exocytosis were affected by complement components, and C-type lectins through intracellular signal transduction pathway. Complement components may involve in the regulation of proPO-activating system to response to ammonia-N stress. This study helped to further understanding the relationship among immune factors of crustacean in response to environmental stress, which implied that when it comes to the decrease of immunity affected by environmental stress, we should not only focus on the mechanism of upstream neuroendocrine response, but also pay attention to the role of immune factors.

The Dairy Cow Slurry Composition Used as Organic Fertilizer Is Influenced by the Level and Origin of the Dietary Protein.

Less than 30% of dairy cattle's nitrogen ingested is retained in milk. Therefore, large amounts of nitrogen can be excreted in manure and urine with a potential environmental impact. In addition, some legume forages can be more susceptible to proteolysis during the silage process than grasses, and dairy cows fed these legume silages would excrete a larger quantity of nitrogen in slurry. The objectives of this work were to evaluate the amount of nitrogen excretion in dairy cows fed different protein levels and legume silages with a view to improve the slurry quality as a co-product that can be used as fertilizer. Two double 3 × 3 Latin square trials were carried out in order to study three different protein levels (high, medium, and low) and three different silages (grass, faba bean, and field pea). Dry matter intake, milk production, and composition were not affected by treatments. The excretion of ammonia-N in the urine was almost four times lower in the diet with the lowest protein level. The ammonia-N in the urine was twice as high with the pea silage than faba bean and grass silages. In conclusion, the diet containing 13% of protein meets the protein requirement for lactating cows producing 31 kg daily, with low nitrogen excretion in the urine, and the main pathway for the excretion of surplus nitrogen from legume silages is through urine and the metabolization of pea silage protein goes toward ammonia-N.

Immune defense, detoxification, and metabolic changes in juvenile Eriocheir sinensis exposed to acute ammonia.

Ammonia-N accumulation in the rice-crab co-culture system may have negative effects on the health of juvenile Eriocheir sinensis. In this study, physiological, transcriptomic, and metabolomic analyses were performed to explore the toxic responses in the hepatopancreas of juvenile E. sinensis exposed to 0, 0.75, and 2.99 mmol/L total ammonia-N for 24 h. We observed that the content of most amino acids and glycogen was significantly decreased after ammonia exposure. Acid phosphatase and alkaline phosphatase activities showed marginally increased trends after low ammonia exposure. Transcriptomic analysis indicated that immune defense, detoxification, and metabolic pathways were altered. Metabolomic analysis revealed that ammonia exposure affected energy metabolism and nucleotide metabolism. The combination of transcriptomic and metabolomic analyses revealed that the tricarboxylic acid cycle and amino acid consumption were enhanced for additional energy supply to cope with ammonia stress. Ammonia stress activated the immune defense system in juvenile E. sinensis. Moreover, the upregulation of detoxification genes and the acceleration of glycogen degradation for glucose supply are important adaptive mechanisms in response to high ammonia stress. Notably, ammonia stress may affect the nervous system of juvenile E. sinensis. Thus, our data provide a better understanding of the defensive mechanisms of E. sinensis against ammonia toxicity.

Metabolic changes and stress damage induced by ammonia exposure in juvenile Eriocheir sinensis.

The application of nitrogen fertilizers in the rice-crab co-culture system may expose juvenile Eriocheir sinensis to high ammonia concentrations within a short period of time, potentially causing death. Currently, the molecular mechanism underlying ammonia toxicity in juvenile Eriocheir sinensis remains poorly understood. This study compared the effects of 24 h exposure to different total ammonia-N concentrations (0, 10.47, and 41.87 mg/L) on antioxidant enzyme activities and tandem mass tag (TMT)-based proteomics in the hepatopancreas of juvenile Eriocheir sinensis. During the experiment, water temperature and pH were maintained at 20.4 ± 1.4 °C and 7.69 ± 0.46, respectively. Proteomic data demonstrated that Eriocheir sinensis used different metabolic regulatory mechanisms to adapt to varying ammonia conditions. The tricarboxylic acid (TCA) cycle, glycogen degradation, and oxidative phosphorylation showed marginally upregulated trends under low ammonia exposure. High ammonia stress caused downregulation of the TCA cycle and provided energy by enhancing oxidative phosphorylation, fatty acid beta oxidation, gluconeogenesis, and glycogen degradation. The detoxification of ammonia into urea and glutamine was suppressed under high ammonia stress. Finally, ammonia exposure induced oxidative stress and caused protein damage. Antioxidant enzyme activity analysis further revealed that exposure to high concentrations of ammonia may induce more severe oxidative stress. This study provides a global perspective on the mechanisms underlying ammonia exposure-induced metabolic changes and stress damage in juvenile Eriocheir sinensis.

The hepatopancreas of the mangrove crab Neosarmatium africanum: a possible key to understanding the effects of wastewater exposure (Mayotte Island, Indian Ocean).

Mangrove crabs are ecosystem engineers through their bioturbation activity. On Mayotte Island, the abundance of Neosarmatium africanum decreased in wastewater-impacted areas. Previous analyses showed that global crab metabolism is impacted by wastewater, with a burst in O2 consumption that may be caused by osmo-respiratory trade-offs since gill functioning was impacted. As the hepatopancreas is a key metabolic organ, the purpose of this study was to investigate the physiological effects of wastewater and ammonia-N 5-h exposure on crabs to better understand the potential trade-offs underlying the global metabolic state. Catalase, superoxide dismutase, glutathione S-transferase, total digestive protease, and serine protease (trypsin and chymotrypsin) activities were assessed. Histological analyses were performed to determine structural modifications. No effect of short-term wastewater and ammonia-N exposure was found in antioxidant defenses or digestive enzyme activity. However, histological changes of B-cells indicate an increase in intracellular digestive activity through higher vacuolization processes and tubule dilation in wastewater-exposed crabs.

Bio-electrochemically extracted nitrogen from residual resources for microbial protein production.

Upcycling of nutrients from residual resources for producing microbial protein (MP) is an attractive method to valorize residues. In this study, we investigated bio-electrochemical methods to recover ammonia-N, for further production of MP. Reject water and digestate were used for ammonia-N recovery in microbial fuel cell (MFC) system. In one-stage process, ammonia-N recovery was 32 - 42% with 57 - 154 kJ/m3 waste stream of electricity generation. For further enhancing recovery efficiency, a two-stage process was developed, achieving efficiency of 53 - 61%. Subsequently, MP was grown with the extracted ammonia-N, and amino acid concentration was 421 and 272 mg/L under 25 °C and 35 °C, respectively. Similar essential amino acid content of MP (especially under 25 °C) with the one from fish demonstrated the attractiveness of upcycling residues to proteins. Based on simplified economic evaluation, the produced energy performed the potential to catch 1.63 - 6.54 €/m3 waste stream.

Treatment of petroleum wastewater contaminated with hydrocarbons and inorganics by anoxic-aerobic sequential moving bed reactors.

The present study deals with the biotransformation of virulent petroleum refinery concoction with phenol (750 mg/L), emulsified crude oil (300 mg/L), S2- (750 mg/L), NH4+-N (350 mg/L) and NO3--N (1000 mg/L) in anoxic (A1) - aerobic (A2) moving bed reactors operated in series. The efficacy of the system was analysed through measurement of pollutant concentrations, GC-MS and FTIR peaks of the influent and effluent, and biomass activity studies. The system was able to eliminate the organics and inorganics with more than 99% efficiency at 80 h HRT and 64 h cycle time. GC-MS results revealed breakage of high molecular weight organics to smaller compounds after anoxic treatment. Further treatment of anoxic effluent by aerobic biomass reduced the number of peaks in the final effluent significantly. FTIR results were in accord with the GC-MS results. Heterotrophic activity (HA) of the aerobic biomass was higher than anoxic biomass due to its higher free energy change. Anoxic biomass showed chemolithotrophic activity (CA), suggesting survival in the absence of organics. Gas generated from anoxic reactor consisted of 91% nitrogen, 1% CO2, 1% H2S and rest was unaccounted.

Orange juice industry by-product silage can increase fat and protein in Holstein cow's milk.

We aimed to evaluate the effect of replacing corn silage by orange peel silage on nutrient intake, ruminal parameters and milk production of multiparous lactating Holstein cows. Eight fistulated Holstein cows averaging 587.5 ± 39.6 kg and 111 ± 22 d in milking were randomly assigned to a double 4 × 4 Latin square design carried out two times to determine the effects of feeding with orange peel silage (OPS) in substitution of whole plant corn silage (WPCS). The treatments were a control diet with WPCS only or diets with OPS replacing WPCS in the total mixed diet (250, 500, or 750 g/kg DM). All cows were fed the same 750 : 250 g/kg roughage : concentrate ratio. The DM intake and milk production were reduced with the OPS inclusion, with decreases in consumption of neutral detergent fibre and increased consumption of non-fibrous carbohydrates. Diets with 250 and 500 g/kg OPS showed similar milk production and protein content in milk to the standard WCPS diet, whilst 750 g/kg orange peel silage as roughage increased fat and protein contents significantly. The orange peel silage as a substitute for corn silage for feeding dairy cows did not show adverse changes in the rumen environment and showed promising results in the increase of fat in milk of Holstein cows.

mTOR - Mediated protein synthesis by inhibiting protein catabolism in Chinese perch (Siniperca chuatsi).

Activation of the mechanistic target of rapamycin (mTOR) pathway is known to promote protein synthesis by enhancing mRNA translation. However, there have been few literatures on the effect of mTOR on protein metabolism in non-mammals. The main source of ammonia in fish comes from protein catabolism. The key step of protein catabolism involves the deamination and/or transamination of amino acids. This study is aimed to explore the mechanism underlying mTOR pathway influencing protein retention from the perspective of protein catabolism. Chinese perch were fasted for 24 h and divided into 4 groups randomly before intracerebroventricular (ICV) injection: (1) control group for leucine; (2) leucine group; (3) control group for leucine and rapamycin; (4) leucine and rapamycin group. Food intake was equivalent between each control and treatment groups at each time point (0.5, 4, 12 and 24 h post-injection). Ammonia-N excretion rate, blood glucose, S6 phosphorylation level, and expression of relative genes of protein catabolism (GDH, AMPD, AST, ALT) were determined. The results indicated that the pS6 level was increased, and that the ammonia-N excretion rate, blood glucose, and mRNA level of protein catabolism genes (GDH and AMPD) were significantly decreased after injection with leucine, while those changes were reversed after injection with leucine and rapamycin. Our study not only reveals the mechanism by which mTOR mediates protein synthesis by inhibiting protein catabolism in Chinese perch, but also provides reference for improving the utilization of feed protein.

Valine acts as a nutritional signal in brain to activate TORC1 and attenuate postprandial ammonia-N excretion in Chinese perch (Siniperca chuatsi).

An emerging concept is that the hypothalamic nutrient sensor can regulate peripheral energy metabolism via a brain-liver circuit. Valine is an essential branched-chain amino acid (BCAA) that drives intracellular signaling cascades by the activation of target of rapamycin complex 1 (TORC1) which is critical to protein metabolism in mammals. However, in teleost fish, it remains scarce in this area especially about how the intraventricular (ICV) injection of valine can mediate the protein metabolism in peripheral organs. This study would tentatively explore the effects of ICV injection of valine on protein metabolism in peripheral organs through evaluating the postprandial ammonia-N excretion rate in Chinese perch. The control group was injected with 5-µL PBS, and the Val group was injected with 20-µg L valine dissolved into 5-µL PBS. The ammonia-N excretion rate of Val group was lower than control group at 4-, 12-, and 24-h postinjection, while the concertation of plasma glucose was increased sharply at 0.5-, 4-, 12-, and 24-h postinjection. We further checked both mRNA level and the enzyme activity of glutamate dehydrogenase (GDH) in the liver and adenosine monophosphate deaminase (AMPD) in muscle, and we found that they were obviously decreased in Val group at 4-, 12-, and 24-h postinjection. The phosphorylation level of ribosomal protein S6, a downstream target protein of TORC1, was markedly enhanced in the liver of Val group at 4-, 12-, and 24-h postinjection. Collectively, these results illustrated that ICV injection of valine can attenuate protein degradation in peripheral organs by depressing the GDH and AMPD enzyme activity; on the other hand, the injected valine can trigger the activation of TORC1 in the liver via a brain-liver circuit and then interdict proteolysis.