Greenhouse gases emissions from aquaculture ponds: Different emission patterns and key microbial processes affected by increased nitrogen loading.

Global aquaculture production is expected to rise to meet the growing demand for food worldwide, potentially leading to increased anthropogenic greenhouse gases (GHG) emissions. As the demand for fish protein increases, so will stocking density, feeding amounts, and nitrogen loading in aquaculture ponds. However, the impact of GHG emissions and the underlying microbial processes remain poorly understood. This study investigated the GHG emission characteristics, key microbial processes, and environmental drivers underlying GHG emissions in low and high nitrogen loading aquaculture ponds (LNP and HNP). The N2O flux in HNP (43.1 ± 11.3 µmol m-2 d-1) was significantly higher than in LNP (-11.3 ± 25.1 µmol m-2 d-1), while the dissolved N2O concentration in HNP (52.8 ± 7.1 nmol L-1) was 150 % higher than in LNP (p < 0.01). However, the methane (CH4) and carbon dioxide (CO2) fluxes and concentrations showed no significant differences (p > 0.05). N2O replaced CH4 as the main source of Global Warming Potential in HNP. Pond sediments acted as a sink for N2O but a source for CH4 and CO2. The △N2O/(△N2O + â–³N2) in HNP (0.015 ± 0.007 %) was 7.7-fold higher than in LNP (0.002 ± 0.001 %) (p < 0.05). The chemical oxygen demand to NO2-N ratio was the most important environmental factor explaining the variability of N2O fluxes. Ammonia-oxidizing bacteria driven nitrification in water was the predominant N2O source, while comammox-driven nitrification and nosZII-driven N2O reduction in water were key processes for reducing N2O emission in LNP but decreased in HNP. The strong CH4 oxidization by Methylocystis and CO2 assimilation by algae resulted in low CH4 emissions and CO2 sink in the aquaculture pond. The Mantel test indicated that HNP increased N2O fluxes mainly through altering functional genes composition in water and sediment. Our findings suggest that there is a significant underestimation of N2O emissions without considering the significantly increased △N2O/(△N2O + â–³N2) caused by increased nitrogen loading.

Exploring organic matter conversion pathway and its effect on nitrogen removal in tidal flow constructed wetlands.

Achieving effective nitrogen removal remains a significant challenge faced by constructed wetlands. Although organic matter is a crucial factor influencing nitrogen removal, little attention has been paid to the impact of organic matter conversion pathways on nitrogen removal in constructed wetlands. Here, we showed that endogenous microorganisms performing carbon internalization could be easily enriched in tidal flow constructed wetlands (TFCWs) under its special rhythmic cycle of anaerobic/aerobic operational mode. Endogenous microorganisms could translate influent carbon sources into intracellular carbons during the anaerobic stage and supply the carbon source for endogenous denitrification after the aerobic stage (rest period). Based on these findings, an innovative combined TFCW and Nitrifying-CW system was developed, and robust total nitrogen (TN) removal (82% on average) was achieved even under carbon source limiting conditions. This performance was a substantial improvement compared to the conventional single bed TFCW with multiple "tides" (corresponding to the multiple contact/rest periods) with TN removal of only 54% on average. Simultaneous nitrification-endogenous denitrification (SNED) was found to be the major nitrogen removal pathway in the proposed system. Compared with classical nitrification-denitrification, simultaneous nitrification-endogenous denitrification brings high nitrogen conversion rates and significantly reduces the demand for oxygen and organic carbon. Furthermore, microbial community analysis indicated that endogenous microorganisms such as Candidatus_Competibacter and Defluviicoccus were successfully enriched, accounting for 50.73% and 3.46% in CW1, and 25.25% and 1.76% in CW2, respectively. Together, these mechanisms allow the proposed system to achieve efficient TN removal.

Selenium attenuated food borne cadmium-induced intestinal inflammation in red swamp crayfish (Procambarus clarkii) via regulating PI3K/Akt/NF-κB pathway.

Selenium (Se), an indispensable micronutrient for living organisms, has been extensively studied for its heavy metal-detoxifying properties in diverse biological systems and tissues. Nevertheless, it is not entirely certain whether Se can effectively protect against Cadmium (Cd)-induced gut inflammation, especially in aquatic animals. In this study, we employed various approaches, including transcriptome profiling, histological examinations, assessment of antioxidant enzyme activities, and analysis of gut microbiota composition to investigate the effects on crayfish growth and intestinal health after exposure to dietary Cd (15 mg kg-1 diet) and Se (15 mg kg-1 diet) individually or in combination for 8 weeks. The results revealed that dietary Cd exposure resulted in reduced body weight and survival rates, along with an increased occurrence of intestinal inflammation. Nevertheless, Se supplementation proved effective in mitigating the adverse effects of Cd on growth and gut health. Se exhibited a remarkable ability to counteract the disruption of gut antioxidant abilities induced by dietary Cd, as evidenced by the observed increases in ROS and MDA contents, decrease in GSH levels, and inhibition of antioxidative enzyme activities. At the concentration of 6 mg kg-1 in the diet, Se was found beneficial for maintaining gut microbiota richness and diversity. Among them, Flavobacterium, Thermomonas, and Chloronema displayed a weak negative correlation with the rate of gut inflammation. Meanwhile, the levels of short chain fatty acids (SCFAs), including acetic acid (AA) and butanoic acid (BA), showed a significant increase in the Se-Cd group compared to the Cd-only group. Furthermore, transcriptome analysis exhibited significant responses of the PI3K/Akt and NF-κB pathways following crayfish exposure to dietary Se and Cd, either separately or in combination. In short, this study provides a new evidence regarding the molecular mechanisms through which Se could regulate the PI3K/Akt and NF-κB pathways, either directly or indirectly via ROS and SCFAs, thereby alleviating Cd-induced gut inflammation in crayfish.

The effect of novel aquaculture mode on phosphorus sorption-release in pond sediment.

The emergence of aquaculture modes has brought considerable changes to the aquaculture landscape and profoundly influenced environmental processes. However, there is limited research on nutrient cycling in emerging aquaculture modes. This study investigated the characteristics and mechanisms of sediment phosphorus (P) sorption-release in traditional earthen pond culture (TEP) and pond-tank culture mode (PTC), which represents novel aquaculture modes. The results showed that under higher nutrient load, the PTC did not show significant differences in nutrient concentration in water and sediments compared to TEP. Although there are no significant differences in overlying water P concentration between the modes throughout the entire aquaculture period, the trends of its variation over time are different, which significantly affected the P sorption-release characteristics of sediment. Additionally, correlation analysis suggested that calcium-bound P and hot NaOH-extractable organic P may affect the sorption-release characteristics of sediment as active P fractions. The change in redox condition caused by enzyme-mediated organic matter decomposition (such as protein and lipids) is also an important reason for sediment P release. However, the P fractions and organic matter content showed no significant differences between the two modes. Sediment microbial analysis showed that TEP exhibited a significant dominance of inorganic P-solubilizing bacteria, especially Actinobacteria and Bacilli classes. PTC had a higher proportion of organic P-solubilizing bacteria, primarily in the Bacteroidia class. The quantitative results of the key functional gene phoD in organic P decomposition also showed that the abundance in PTC was significantly higher than that in TEP. This suggested that microbial differences may be another reason for differences in P sorption-release behavior. This study revealed the differences in P sorption-release characteristics and mechanisms between the TEP and PTC, which holds positive implications for water quality and pollution management in novel aquaculture modes.

Mangiferin and Taurine Ameliorate MSRV Infection by Suppressing NF-κB Signaling.

The emergence or reemergence of viruses pose a substantial threat and challenge to the world population, livestock, and wildlife. However, the landscape of antiviral agents either for human or animal viral diseases is still underdeveloped. The far tougher actuality is the case that there are no approved antiviral drugs in the aquaculture industry, although there are diverse viral pathogens. In this study, using a novel epithelial cell line derived from the brain of Micropterus salmoides (MSBr), inflammation and oxidative stress were found to implicate the major pathophysiology of M. salmoides rhabdovirus (MSRV) through transcriptome analysis and biochemical tests. Elevated levels of proinflammatory cytokines (interleukin-1ß [IL-1ß], IL-6, IL-8, tumor necrosis factor alpha [TNF-α], and gamma interferon [IFN-γ]) and accumulated contents of reactive oxygen species (ROS) as well as biomarkers of oxidative damage (protein carbonyl and 8-OHdG) were observed after MSRV infection in the MSBr cells. Mangiferin or taurine dampened MSRV-induced inflammation and rescued the oxidative stress and, thus, inhibited the replication of MSRV in the MSBr cells with 50% effective concentration (EC50) values of 6.77 µg/mL and 8.02 µg/mL, respectively. Further, mangiferin or taurine hampered the activation of NF-κB1 and the NF-κB1 promoter as well as the increase of phosphorylated NF-κB (p65) protein level induced by MSRV infection, indicating their antiviral mechanism by suppressing NF-κB signaling. These findings exemplify a practice approach, aiming to dampen and redirect inflammatory responses, to develop broad-spectrum antivirals. IMPORTANCE Aquaculture now provides almost half of all fish for human food in 2021 and plays a significant role in eliminating hunger, promoting health, and reducing poverty. There are diverse viral pathogens that decrease production in aquaculture. We developed a novel epithelial cell line derived from the brain of Micropterus salmoides, which can be used for virus isolation, gene expressing, and drug screening. In this study, we focus on M. salmoides rhabdovirus (MSRV) and revealed its pathophysiology of inflammation and oxidative stress. Aiming to dampen and redirect inflammatory responses, mangiferin or taurine exhibited their antiviral capability by suppressing NF-κB signaling. Our findings exemplify a practice approach to develop broad-spectrum antivirals by dampening and redirecting inflammatory responses.

BJTJ-1837, a novel FXI activation-blocking antibody.

Background: Factor (F)XI contributes to thrombosis development while it plays a limited role in normal hemostasis. FXI targeting has the potential for preventing and treating thrombosis with little bleeding risk. Objectives: The aim of this study was to develop novel antibody therapeutics against FXI for the treatment of thrombosis-related diseases. Methods: Mouse hybridoma technology was applied to screen for anti-FXI antibodies. Surface plasma resonance, enzyme inhibition, activated partial thromboplastin time, and prothrombin time assays were conducted to characterize the binding affinity and activity of antibodies. A cynomolgus monkey arterial venous shunt model was applied to validate the antithrombotic activities. Results: A humanized antibody, BJTJ-1837, reported here bound to the protease domain of FXI and activated FXI with high affinity. BJTJ-1837 fully inhibited the activation of FXI by activated FXII and thrombin. BJTJ-1837 also demonstrated strong anticoagulant activity in human and cynomolgus monkey plasma as measured by activated partial thromboplastin time. Moreover, BJTJ-1837 showed favorable antithrombotic activity with a dose-dependent protection in an arterial venous shunt thrombosis model in cynomolgus monkeys without the bleeding adverse effect. Furthermore, BJTJ-1837 displayed favorable pharmacokinetic and pharmacodynamic properties and good developability. Conclusion: As a potential antithrombotic therapeutic agent with a safe profile, BJTJ-1837 is a very promising FXI activation-blocking antibody candidate.

Formation of microorganism-derived dissolved organic nitrogen in intermittent aeration constructed wetland and its stimulating effect on phytoplankton production: Implications for nitrogen mitigation.

To control eutrophication in aquatic ecosystems, enhancing nitrogen removal in the constructed wetland (CW) by upgrading conventional CW to aeration CW is commonplace. However, regulatory efforts have only focused on reducing dissolved inorganic nitrogen (DIN) discharge and disregarding dissolved organic nitrogen (DON). Here, we used experimental mesocosms to investigate the effect of aeration mode on the characteristics of effluent DON in CW. The results showed that intermittent aeration is prone to introduce large amounts of DON and bioavailable DON (ABDON) in the effluents, although it greatly decreases effluent total nitrogen (TN). Analysis of DON fluorescent components and molecular characteristics indicated that suddenly shifting the environment from anoxic condition to aerobic condition in the intermittent aeration CW (IACW) would stimulate microorganisms to release tryptophan and simple aromatic proteins-like substances, which does not occur in the limited continuous aeration CW (CACW). Consequently, the abundance of DON resembling lipids, proteins/amino sugars, and carbohydrates-like molecules in IACW were about 2.1 times higher than that in CACW. Bioassay results showed that Selenastrum capricornutum and Microcystis aeruginosa incubated with effluent from IACW both generate larger phytoplankton biomass than that with CACW effluent, even though IACW effluent contains less TN than its counterpart. Moreover, Microcystis aeruginosa can simultaneously utilize DON and DIN, while Selenastrum capricornutum seem to utilize the DON only when DIN was not available. This result implies that increasing DON discharge may also influence phytoplankton composition and stimulate harmful phytoplankton species. Overall, this study indicates that upgrading CW solely depending on DIN removal level cannot ensure a mitigation of nitrogen-related eutrophication, and more efforts should be paid to curb DON discharge.

Sediment diffusion is feasible to simultaneously reduce nitrate discharge from recirculating aquaculture system and ammonium release from sediments in receiving intensive aquaculture pond.

Nitrogen accumulation has become one of the greatest unresolved challenges restricting the development of aquaculture worldwide. In recirculating aquaculture system (RAS), lack of organic matter (OM) and sensitive organisms makes it difficult to apply efficient denitrifying technology, thus leading to a high nitrate­nitrogen (NO3--N) accumulation. In contrast, excess OM accumulation in intensive aquaculture pond sediments is associated with dissolved oxygen depletion and ammonium­nitrogen (NH4+-N) accumulation in the sediments. Based on the opposing effects of OM on the nitrogen accumulation in RAS and intensive aquaculture ponds, this study assessed the feasibility of simultaneously reducing NO3--N discharge from RAS and controlling NH4+-N accumulation in intensive aquaculture ponds by in situ diffusing RAS tailwater containing NO3--N into intensive aquaculture pond sediments. The results showed that NO3--N diffusion strategy improved the native sediment denitrification capacity, thus increasing NO3--N removal efficiency from RAS tailwater and significantly decreasing the NH4+-N concentration in interstitial water and the total organic carbon content in intensive aquaculture pond sediments. High-throughput sequencing and quantitative real-time polymerase chain reaction (qPCR) results revealed that NO3--N addition significantly increased both nitrifying bacteria and denitrifying bacteria abundance. These results implied that NO3--N diffusion strategy could effectively stimulate microbial decomposition of OM, thus relieving the hypoxia limitation of sediment nitrification. Overall, this study offers a feasible method for simultaneous reduction of NO3--N from RAS tailwater and NH4+-N in intensive aquaculture ponds with low cost and high efficiency.

Isoandrographolide from Andrographis paniculata ameliorates tubulointerstitial fibrosis in ureteral obstruction-induced mice, associated with negatively regulating AKT/GSK-3ß/ß-cat signaling pathway.

Tubulointerstitial fibrosis (TIF) is a prominent pathological manifestation for the progression of almost all chronic kidney diseases (CKDs) to end-stage renal failure. However, there exist few efficient therapies to cure TIF. Our recent results showed that (8R, 12S)-isoandrographolide (ISA), a diterpenoid lactone ingredient of traditional Chinese herbal Andrographis paniculata (Burm.f.) Nees, exhibited anti-pulmonary fibrosis in silica-induced mice. Herein, we investigated the therapeutic effect of ISA on TIF, using mice subjected to unilateral ureteral obstruction (UUO) and human kidney proximal tubular epithelial (HK-2) cells treated with transforming growth factor-ß1 (TGF-ß1) or tumor necrosis factor-α (TNF-α). The pathological changes and collagen deposition results displayed that ISA administration significantly attenuated inflammatory response, ameliorated TIF, and protected the kidney injury. Interestingly, ISA revealed much lower cytotoxicity on HK-2 cells, but exhibited stronger inhibitory effect on tubular epithelial mesenchymal transformation (EMT) and inflammation, as compared to andrographolide (AD), the major ingredient of A. paniculata extract that has been reported to ameliorate TIF in diabetic nephropathy mice. It was further clarified that the amelioration of TIF by ISA was associated with suppressing the aberrant activation of AKT/GSK-3ß/ß-catenin pathway through network pharmacology analysis and experimental validation. Taken together, these findings indicate that ISA is a promising lead compound for development of anti-TIF, and even broad-spectrum anti-fibrotic drugs.

Strengthening anoxic glycogen consumption in SNEDPR-CW as a strategy to control PAO-GAO competition under carbon limited condition.

Cooperation between Phosphate and Glycogen Accumulating Organisms (PAOs and GAOs) plays a pivotal role in nutrients removal in simultaneous nitrification endogenous denitrification and phosphorous removal (SNEDPR) systems. Recent findings have expanded the application of SNEDPR from activated sludge system to constructed wetland (CW). However, how to regulate competition between PAOs and GAOs in SNEDPR-based CW is still unclear. Here we showed that, GAOs could easily gain dominance over PAOs in SNEDPR-CW under alternating anaerobic/aerobic (A/O) operational mode. Shortening aerobic hydraulic retention time (HRT) at low oxygen concentration was benefit for simultaneous nitrification endogenous denitrification (SNED) and denitrifying dephosphatation but would reduce the overall phosphorus uptake rate and lead to high phosphorus effluent concentrations. Extended aerobic HRT promoted the proliferation of aerobic GAOs over PAOs, decreasing both enhanced biological phosphorus removal (EBPR) and SNED performance. Surprisingly, by switching the operation of system to alternating anaerobic/aerobic/anoxic (A/O/A) mode, an extraordinary nutrients removal performance with mean nitrogen and phosphorus removal efficiency of 84.57% and 89.37% was achieved under carbon sources limited condition. Stoichiometric analysis demonstrated that adding anoxic stage strengthened the intracellular glycogen oxidization of GAOs for denitrification which compromised its subsequent anaerobic carbon sources uptake and PHA storage and provided sufficient carbon sources for PAOs. Microbial community analysis showed that numerical ratio of GAOs to PAOs decreased from 6.67 under A/O to 4.89 under A/O/A mode, which further indicated strengthening glycogen denitrification of GAOs should be an effective way to regulate microbial competition in order to obtain a desired nutrients removal performance in SNEDPR-CW.

Integrating Microbial Protein Production and Harvest Systems into Pilot-Scale Recirculating Aquaculture Systems for Sustainable Resource Recovery: Linking Nitrogen Recovery to Microbial Communities.

In aquaculture, it is important to raise the nitrogen recovery efficiency (NRE) to improve sustainability. To achieve this, recovery of microbial protein (RMP), instead of nitrification/denitrification in conventional wastewater treatment, is a promising approach whose microbiological mechanisms must be characterized. Here, periodic RMP was conducted in an in situ biofloc-based aquaculture system (IBAS) and a separating assimilation reactor-based recirculating aquaculture system (SRAS). Kinetic analysis indicated that a microbial biomass level of 3 g L-1 was optimal for inorganic N removal, and excess biomass was harvested to improve the NRE. Unlike the IBAS, the SRAS eliminated the fluctuation in water quality caused by the RMP. Periodic RMP significantly increased the NRE to 44-57% by promoting the filamentous bacterium Herpetosiphon and suppressing anaerobic denitrifiers. Aerobic chemoheterotrophy was the main microbial metabolic process for energy. After RMP, nitrate reductase-encoded functional genes (napA and narG) significantly decreased, while nitrite reductase-encoded functional genes, especially nirK, significantly increased. Co-occurrence networks analysis indicated that the cooperation and competition among organic matter degraders, filamentous bacteria, nitrifiers, and denitrifiers determined the microbial protein yield. These results provide fundamental insights into the influence of the RMP on microbial communities and functions, which is important for realizing sustainable aquaculture.

Advanced nitrogen and phosphorus removal by combining endogenous denitrification and denitrifying dephosphatation in constructed wetlands.

To achieve high-efficiency nutrient removal in constructed wetlands (CWs), a novel simultaneous nitrogen and phosphorus removal (SNPR) process was developed by combining nitrification, endogenous denitrification, and denitrifying phosphorus removal. In SNPR process, denitrifying glycogen-accumulating organisms (DGAOs) and denitrifying polyphosphate-accumulating organisms (DPAOs) utilized NOx--N(NO3--N or NO2--N) as electron acceptor and poly-beta-hydroxy-alkanoates (PHAs) as carbon sources for endogenous denitrification and denitrifying phosphorus removal processes. Results from 217 days of operation showed that a high-level of nitrogen removal efficiency of 83.73% was achieved with influent COD/N of 4. The success was attributed to the fact that most of influent carbon sources could be transformed into PHAs before nitrification via enriching DGAOs and DPAOs in CW, which simultaneously improved nitrification and denitrification due to reducing oxygen and carbon sources consumption by aerobic heterotrophs. Phosphorus was mainly removed via denitrifying phosphorus removal, and PO43--P removal efficiency reached up to 87.84% with even common gravel used as substrate. Stoichiometry analysis revealed that DGAOs were the main organisms providing nitrite to DPAOs, suggesting that the effective PO43--P removal under high DGAO abundance condition might be attributed to the coordination of DGAOs and DPAOs in SNRP processes.

Dynamic changes in microbial community structure in farming pond water and their effect on the intestinal microbial community profile in juvenile common carp (Cyprinus carpio L.).

Water quality parameter dynamics, gut, sediment and water bacteria communities were studied to understand the environmental influence on the gut microbial community of a new strain of Huanghe common carp. A total of 3,384,078 raw tags and 5105 OTUs were obtained for the gut, water and sediment bacteria. The water quality had a stronger influence on the water bacteria community than gut and sediment bacteria communities. The ambient water quality parameters also significantly influenced the water and sediment bacteria communities. Comparing the gut, sediment, and water microbial communities, a relationship was found among them. However, gut bacteria were more closely related to sediment bacterial communities than to water bacteria communities. The results showed that the top three bacterial taxa were identical in gut and sediment samples in the early days of rearing. Interestingly, bacterial communities in the carp gut, water, and sediment had different adaptabilities to variations in environmental factors.

Nitrogen removal by heterotrophic nitrifying and aerobic denitrifying bacterium Pseudomonas sp. DM02: Removal performance, mechanism and immobilized application for real aquaculture wastewater treatment.

A bacterial strain was isolated and identified as Pseudomonas sp. DM02 from an aquaculture system. Strain DM02 showed efficient heterotrophic nitrification-aerobic denitrification capability. Total ammonia nitrogen (TAN, 10 mg/L) could be completely removed by strain DM02 within 12 h under low nutrient condition. Nitrogen mass balance indicated that 70.8% of the initial TAN was translated into gaseous nitrogen and 28.1% was converted into intracellular nitrogen. Various carbon sources can be used for nitrate removal (>95% within 28 h). The optimal conditions for TAN, nitrate and nitrite removal were pH 7 with carbon/nitrogen (C/N) ratios of 8, 12 and 12, respectively. The napA, nirK, and nosZ functional genes were successful amplified from strain DM02. Both bioaugmentation and immobilized technology of strain DM02 present ability (>88%) for continuous treatment of real aquaculture wastewater. This research indicated a great potential for practical application of Pseudomonas sp. DM02 in aquaculture wastewater treatment.

Aerobic Denitrification Microbial Community and Function in Zero-Discharge Recirculating Aquaculture System Using a Single Biofloc-Based Suspended Growth Reactor: Influence of the Carbon-to-Nitrogen Ratio.

In this study, the effect of aerobic denitrification on nitrogen removal was investigated using two zero-discharge biofloc-based recirculating aquaculture systems with representative carbon-to-nitrogen (C/N) ratios of 15 (CN15) and 20 (CN20). Aquaculture wastewater, residual feed, and fish feces were treated in an aerated suspended growth reactor (SGR, dissolved oxygen > 5.0 mg L-1). Low toxic NH3 (<0.1 mg L-1) and NO2 --N (<0.5 mg L-1) concentrations and high NO3 --N (83.3%) and NO2 --N (100%) removal efficiencies were achieved in the fish tank and SGR of CN20, respectively. The nitrogen mass balances indicated that the gaseous nitrogen loss accounted for 72-75% of the nitrogen input. Illumina sequencing and quantitative polymerase chain reaction revealed that increasing the C/N ratio significantly increased the amount of aerobic denitrifying bacteria (Dechloromonas, Rhodobacter, Flavobacterium, and Zoogloea) and aerobic denitrifying functional genes (napA, nirK, and nosZ). Autotrophic Nitrosomonas was the dominant nitrifying bacteria in the CN15 system, and autotrophic (Nitrosomonas) and heterotrophic nitrifiers coexisted in the CN20 system. Moreover, the functional prediction analysis showed that the carbohydrate, energy, and amino acid metabolisms in the SGR of the latter increased. In conclusion, aerobic denitrification should widely exist in biofloc systems.

Improvement of Muscle Quality of Grass Carp (Ctenopharyngodon idellus) With a Bio-Floating Bed in Culture Ponds.

Muscle quality and the physiological condition of fish are affected greatly by the culture environment. In aquaculture ponds, a bio-floating bed planted with vegetation is often used to improve water quality. This study investigated the growth and muscle quality of grass carp (Ctenopharyngodon idellus) cultured in ponds equipped with bio-floating beds. Fish were cultured in two replicated pond groups from May to November. Fish in the first group were cultured in experimental ponds equipped with a bio-floating bed planted with Ipomoea aquatica, whereas fish in the other group were reared in control ponds without a bio-floating bed. Compared with control ponds, the experimental ponds had better water quality with significantly lower concentrations of nitrite and ammonia. Grass carp in the experimental group had greater muscle weight gain, a significantly higher content of crude protein, and a significantly lower crude fat level than fish in the control group. The levels of pH, water-holding capacity, and antioxidant capacity of muscle decreased significantly in the control group compared to the experimental group. Texture profile analysis revealed higher values of hardness, springiness, gumminess, and chewiness and lower values of cohesiveness and resilience of white muscle in the experimental group compared to the control group. The filets of fish in the experimental group also received higher grades in the sensory evaluation of springiness, overall acceptability, aroma, and palatability. These results indicate that growth performance and muscle quality of grass carp were improved by the presence of bio-floating beds in the culture ponds.

Diet Affects Muscle Quality and Growth Traits of Grass Carp (Ctenopharyngodon idellus): A Comparison Between Grass and Artificial Feed.

Fish muscle, the main edible parts with high protein level and low fat level, is consumed worldwide. Diet contributes greatly to fish growth performance and muscle quality. In order to elucidate the correlation between diet and muscle quality, the same batch of juvenile grass carp (Ctenopharyngodon idellus) were divided into two groups and fed with either grass (Lolium perenne, Euphrasia pectinata and Sorghum sudanense) or artificial feed, respectively. However, the different two diets didn't result in significant differences in all the detected water quality parameters (e.g., Tm, pH, DO, NH3/[Formula: see text]-N, [Formula: see text]-N, [Formula: see text], TN, TP, and TOC) between the two experimental groups. After a 4-month culture period, various indexes and expression of myogenic regulatory factor (MRFs) and their related genes were tested. The weight gain of the fish fed with artificial feed (AFG) was nearly 40% higher than the fish fed with grass (GFG). Significantly higher alkaline phosphatase, total cholestrol, high density cholestrol and total protein were detected in GFG as compared to AFG. GFG also showed increased hardness, resilience and shear force in texture profile analysis, with significantly bigger and compact muscle fibers in histologic slices. The fat accumulation was most serious in the abdomen muscle of AFG. Additionally, the expression levels of MyoG, MyoD, IGF-1, and MSTNs were higher, whereas Myf-5, MRF4, and IGF-2 were lower in most positional muscles of GFG as compared to AFG. Overall, these results suggested that feeding grass could promote muscle growth and development by stimulating muscle fiber hypertrophy, as well as significantly enhance the expression of CoL1As. Feeding C. idellus with grass could also improve flesh quality by improving muscle characteristics, enhancing the production of collagen, meanthile, reducing fat accumulation and moisture in muscle, but at the cost of a slower growth.

Enhancing Nitrate Removal from Freshwater Pond by Regulating Carbon/Nitrogen Ratio.

Nitrogen accumulation is a serious environmental problem in freshwater ponds, which can lead to massive death of fish and shrimps as well as the eutrophication. The removal of nitrate by regulating the carbon to nitrogen (C/N) ratio and the underlying mechanisms were investigated. The nitrate removal system comprised 530-mL medium containing 5 mg/L [Formula: see text]-N and 0-66.6 mg/L COD (i.e., C/N ratio of 0-13.3) and 20 g ponds sediments. When the C/N ratio was higher than 8, the nitrate removal efficiency nearly reached 100% during the incubation period and the accumulation of nitrite was negligible. When the C/N ratio was below 8, the nitrate removal efficiency was lower and significant nitrite accumulation occurred. The nitrate removal rate increased with the C/N ratio increased, which was ascribed to the increase in the absolute abundance of denitrifiers (nirS, nirK, and nosZ). Although both nirS-type and nirK-type denitrifiers were found in the sediments of freshwater pond, nirS-type denitrifiers were predominant. Dechloromonas was the major nirS-type denitrifier for nitrate removal in nirS-type with the C/N ratios above 5.33, while the majority of the nirK-type denitrifiers were unclassified. Thus, this study implied that the appropriate C/N ratio played an important role on the removal of excess nitrate from freshwater ponds.

Achieving short-cut nitrification and denitrification in modified intermittently aerated constructed wetland.

This study aim to enhance nitrogen removal performance via shifting nitrogen removal pathway from nitrate to nitrite pathway. It was demonstrated that nitrite pathway was successfully and stably achieved in CWs by using modified intermittent aeration control with aeration 20min/non-aeration 100min and reducing DO concentration during aeration, nitrite in the effluent could accumulate to over 70% of the total oxidized nitrogen. Q-PCR analysis showed that nitrifying microbial communities were optimized under the alternating anoxic and aerobic conditions, ammonia oxidizing bacteria increased from 7.15×106 to 8.99×106copies/g, while nitrite oxidizing bacteria decreased approximately threefold after 234days operation. Most importantly, high nitrogen removal efficiency with ammonium removal efficiency of 94.6%, and total nitrogen removal efficiency of 82.6% could be achieved via nitrite pathway even under carbon limiting conditions. In comparison to the nitrate pathway, the nitrite pathway could improve the TN removal by about 55%.

Bacterial abundance and diversity in pond water supplied with different feeds.

The abundance and diversity of bacteria in two types of ponds were investigated by quantitative PCR and Illumina MiSeq sequencing. The results revealed that the abundance of bacterial 16S rRNA genes in D ponds (with grass carp fed sudan grass) was significantly lower than that in E ponds (with grass carp fed commercial feed). The microbial communities were dominated by Proteobacteria, Cyanobacteria, Bacteroidetes, and Actinobacteria in both E and D ponds, while the abundance of some genera was significantly different between the two types of ponds. Specifically, some potential pathogens such as Acinetobacter and Aeromonas were found to be significantly decreased, while some probiotics such as Comamonadaceae unclassified and Bacillales unclassified were significantly increased in D ponds. In addition, water quality of D ponds was better than that of E ponds. Temperature, dissolved oxygen and nutrients had significant influence on bacterial communities. The differences in bacterial community compositions between the two types of ponds could be partially explained by the different water conditions.