Occurrence, distribution and sources of microplastics in typical marine recirculating aquaculture system (RAS) in China: The critical role of RAS operating time and microfilter.

Industrial mariculture, a vital means of providing high quality protein to humans, is a potential source of microplastics (MPs) which have recently received increasing attention. This study investigated the occurrence and distribution of microplastics in feed, source water and recirculating aquaculture system (RAS) with long & short operating times as well as in fish from typical industrial mariculture farms in China. Results showed that microplastics occurred in all samples with the average concentration of 3.53 ± 1.39 particles/g, 0.70 ± 0.17 particles/L, 1.53 ± 0.21 particles/L and 2.21 ± 0.62 particles/individual for feed, source water, RAS and fish, respectively. Microplastics were mainly fiber in shape, blue in color and 20-500 µm in size. Compared with short operated RAS, long operating time led to higher microplastic concentration in RAS, especially that of microplastic in 20-500 µm, granular and blue. Regardless of short or long operating time, microplastics in RAS mainly gathered in culture tank, tank before microfilter and fixed-bed biological filter, and the microfilter removed efficiently the microplastic with the shape of film, granule, fragment as well as those with size > 1000 µm. As for the polymer types, polyamide (PA, 71.9 %) and polyethylene terephthalate (PET, 65.7 %) dominated in feed and source water, respectively, which may be the reason for the high proportion of PA (38.8 % and 26.4 %) and PET (31.8 % and 30.2 %) in RAS and fish. In addition, polypropylene (PP) was also detected in RAS (18.7 %) and fish (22.6 %), indicating that other plastic facilities such as PP brush carrier also made a contribution. Positive matrix factorization (PMF) model revealed three sources of MP in RAS, namely plastic facilities, industrial sewage and plastic packaging products. Our results provided a theoretical basis for the management of MP in RAS.

Gut microbiota dynamics interacting with gastrointestinal evacuation of Apostichopus japonicus: novel insights into promising strategies for environmental improvement.

As an important input of environmental micropollutants into aquaculture environment, feed is now considered to be a critical factor in shaping gastrointestinal evacuation characteristics of animals. We analyzed the gastrointestinal evacuation characteristics and gut bacteria of Apostichopus japonicus within 30 h after feeding in recirculating aquaculture system (RAS) and explored the evacuation mechanism interacting by bacteria. The Gauss model was the most precise gastrointestinal evacuation curve, and 80% of gastrointestinal evacuation time was 27.81 h after feeding. Linear discriminant analysis effect size analysis revealed that gut microbial abundance associated significantly with time (P < 0.05), and 42 biomarkers that could predict gastrointestinal evacuation were totally detected, such as Lutibacter and Vibrio. Biomarkers at 25 h after feeding were related to harmful bacteria. A dynamic response between gastrointestinal content ratio and gut microbial abundance was detected. Taken together, we could discharge sewage about 25 h after feeding and carry out the next round of feeding activities.

Enhanced denitrification of biodegradable polymers using Bacillus pumilus in aerobic denitrification bioreactors: Performance and mechanism.

Nitrate accumulation is an important issue that affects animal health and causes eutrophication. This study combined biodegradable polymers with degrading bacteria to lead to high denitrification efficiency. The results showed polycaprolactone had the highest degradation and carbon release rate (0.214 mg/g∙d) and nitrogen removal was greatest when the Bacillus pumilus and Halomonas venusta ratio was 1:2. When the hydraulic retention time was extended to 12 h, the nitrate removal rate for H. venusta with B. pumilus and polycaprolactone increased by 48 %. Furthermore, the group with B. pumilus contained more Proteobacteria (77.34 %) and denitrifying functional enzymes than the group without B. pumilus. These findings indicated B.pumilus can enhance the degradation of biodegradable polymers especially polycaprolactone to improve the denitrification of the aerobic denitrification bacteria H.venusta when treating maricultural wastewater.

Study on modified SA-H3BO3 immobilization microorganism method for wastewater treatment in seawater recirculating aquaculture system.

The sodium alginate-H3BO3 (SA-H3BO3) is traditionally used as bioremediation method for wastewater treatment in recirculating aquaculture system. Even though this method has many advantages (e.g., high cell loading) for immobilization, the remove of ammonium is not very effective. In this study, a modified method was built by adding polyvinyl alcohol and activated carbon into SA solution, and then crosslinked with saturated H3BO3-CaCl2 solution for creating new beads. Moreover, response surface methodology was utilized for optimizing the immobilization based on Box-Behnken design. The removal rate of ammonium in 96 h was taken as the primary performance criterion to characterize the biological activity of immobilized microorganisms (i.e., Chloyella pyrenoidosa, Spirulina platensis, Nitrifying bacteria, and Photosynthetic bacteria). Based on the results, the optimal parameter of immobilization as follows: the concentration of SA was 1.46%, the concentration of polyvinyl alcohol was 0.23%, the concentration of activated carbon was 0.11%, the crosslinking time was 29.33 h, and the pH was 6.6.

Interaction among biofilter microbiome, fecal metabolome and water quality and regulation of sewage discharge in the recirculating aquaculture system of Apostichopus japonicus.

With the aggravation of environmental pollution caused by traditional culture of Apostichopus japonicus, the concept of A. japonicus recirculating aquaculture system (RAS) came into being. To plan the sewage discharge time reasonably, we explored the temporal variation of water quality, biofilter microbe and fecal metabolome in RAS and relevant mechanism. The results showed that monitored water quality in RAS were within the safe living range of A. japonicus. Proteobacteria and Desulfobacterota were dominant bacteria in biofilter. The RDA results and correlation heatmap showed that NH4-N and NO2-N significantly affected the microbial community composition. The expression pattern of fecal metabolites changed with the passage of time after feeding. And ROC curve analysis and VIP bar chart showed that there were inter group biomarkers with predictive performance, which could help to remind timely sewage discharge. Topological analysis of KEGG pathway enrichment showed that metabolic pathways such as alanine, aspartate and glutamate metabolism changed significantly after feeding (P < 0.01). Additionally, the correlation analysis results showed that biofilter microbe and fecal metabolites were related to water quality (P < 0.05). Combined with the above research results, this study concluded that the RAS could discharge sewage 25-30 h after feeding. These findings were of direct significance to the management of RAS environment and the protection of A. japonicus healthy growth.

Effects of light intensity and photoperiod on the cultivation of the soft coral Sarcophyton trocheliophorum.

Coral reefs are one of the most diverse, productive ecosystems in the world, and light plays crucial role in its survival. Notably, the effects of light conditions on soft coral and its adaptive mechanism were unclear. Thus, the present study aimed to investigate and evaluate the effects of different light intensities (30, 80 and 130 µmol m-2 s-1) and photoperiods (18D:6L, 12D:12L and 6D:18L) on cultivation of soft coral Sarcophyton trocheliophorum. During two 50-day of the experiments, we monitored the zooxanthellae density, Chl a content, enzyme activities (SOD, CAT and GST) and microbial diversity of S. trocheliophorum. Our study's outcomes found that, at the end of the experiment, the 80 µmol m-2 s-1 light intensity group and 12D:12L photoperiod group both possessed the highest zooxanthellae density (2.54 × 108 ± 0.14 × 108 cells g-1 DW and 2.40 × 108 ± 0.07 × 108 cells g-1 DW, respectively), Chl a content (295.01 ± 14.13 µg g-1 DW and 287.78 ± 16.13 µg g-1 DW, respectively) and microbial diversity and relatively stable enzyme activities level. Besides, we speculated that the reason for the decline of zooxanthellae density, Chl a content and microbial diversity under other light conditions might be that it induced light stress and caused oxidative damage. The main bacterial composition of S. trocheliophorum in different light conditions was similar at the phylum level, showing the stability of microbial community structure. Proteobacteria, Actinobacteria and Firmicutes were dominant under all light conditions, so we hypothesized that these bacteria phylum play a crucial role in coral growth and survival. In conclusion, compared with the other treatments, 80 µmol m-2 s-1 light intensity and 12D:12L photoperiod were more beneficial to the growth performance of S. trocheliophorum and could be recommended for its cultivation condition. Our study could provide helpful information for sustainable management plans for the cultivation and conservation of soft corals, which was especially important to the protection and restoration of degraded coral reefs.

Acute low-dose phosphate disrupts glycerophospholipid metabolism and induces stress in juvenile turbot (Scophthalmus maximus).

Phosphate, as the main nutrient factor of lake eutrophication brought by pollutants discharged from agriculture and industry, is always considered to be a low-toxicity substance to aquatic animals. But the toxicity mechanism is unclear, and published information is limited. In this study, a 96 h acute stress experiment was conducted on juvenile turbot (Scophthalmus maximus) with 0, 10, and 60 mg/L phosphate solutions. Metabonomic analysis revealed that low-dose phosphate (10 mg/L) disrupted glycerophospholipid, purine, and glycolipid metabolism, as well as the tricarboxylic acid (TCA) cycle in juveniles, even at 96 h of stress, which may lead to cell structure damage and signal recognition disorder between cells. Upregulated key genes in the main glycerophospholipid metabolic pathways, which matched the results of the metabolomic study, were detected. Furthermore, low-dose phosphate (10 mg/L) induced oxidative stress and immunotoxicity in fish, resulting in the raising of relevant genes expression such as cat and sod in liver and kidney. In addition, all phosphate-treated groups had induced lesions on gill tissue, as evidenced by pathological observations. In this study on toxic effects on and mechanism of phosphate in aquatic animals using metabolomics, gene expression, and histopathology, we confirm that acute low-dose phosphate could disrupt glycerophospholipid metabolism and induce stress in juvenile turbot. This can provide advice on the amount of phosphate accumulation for marine fish farming and on protecting species diversity and marine ecosystem from the point of view of phosphate toxicity to marine animals.

Carbon sources derived from maize cobs enhanced nitrogen removal in saline constructed wetland microcosms treating mariculture effluents under greenhouse condition.

This study investigated an alternative carbon source derived from maize cobs (MCs) to enhance nitrogen removal in saline constructed wetlands (SCWs). The main objectives were to select the proper pretreatment method of MCs for rapid carbon release; and to investigate the effects of maize cob pieces (i.e. MCP) and three addition levels of maize cob lixiviums (i.e. L-MCL, M-MCL and H-MCL) on nitrogen purification performance and microbial characteristics of SCWs. Results showed NaOH pretreatment enhanced carbon release of MCs in seawater (from 7.5 ± 0.4 mgCOD g-1 to 16.4 ± 0.2 mgCOD g-1). The 80-d trial showed SCWs with M-MCL addition performed well on nitrogen removal: NO3-N, 88.8 ± 11.6%; NO2-N, 91.1 ± 3.5%; TAN, 96.5 ± 1.6%; TIN, 89.8 ± 10.4%; with 2 mg L-1 effluent COD. Denitrification parameters confirmed MCL to be a high quality carbon source: denitrification potential (PDN) = 0.16 gN gCOD-1; heterotrophy anoxic yield coefficient (YH) = 0.54 gCOD gCOD-1. The MCP and H-MCL treatments improved substrate dehydrogenase activity, indicating a higher microbial activity in these SCWs. Sequencing analysis revealed that, regardless of addition manners, carbon sources from MCs changed the rhizosphere microbial community. At genus level, Anaerophaga (10.1%), Granulosicoccus (8.2%) and Sulfurimonas (6.6%) dominated in SCWs under MCP treatment. Increased MCL addition levels improved the relative abundance of Vibrio, Malonomonas and Caldithrix, suggesting the enhancement of denitrification. Relative high proportions of Desulfotignum and Desulfovibrio, and Sulfurimonas were observed in MCP and H-MCL SCWs, implying that sulfate reduction occurred in SCWs with excess carbon sources.

Denitrification characteristics and pathways of a facultative anaerobic denitrifying strain, Pseudomonas denitrificans G1.

Excessive nitrate in aquaculture systems has attracted wide attention. To isolate novel aerobic denitrifying strain and characterize its nitrogen removal processes, a facultative anaerobic denitrification bacterium, identified as Pseudomonas denitrificans G1, was isolated from marine sediments. Strain G1 could grow and remove 90-98% of nitrate and 97-99% of nitrite under an aerobic or anaerobic condition in 24 h, with the total nitrogen removal rate of 33-38% (87-100 mg/L). The highest denitrification rate could reach 15.1 mg/(L·h). The suitable condition for the denitrification of G1 is C/N ratio 5-22, dissolved oxygen 0-4.68 mg/L, salinity 0-30 g NaCl/L, pH 7-9.5. Under the aerobic condition, G1 grew fast; however, the mass spectrographic analysis showed that the gas product was N2O. Under the anaerobic conditions, G1 grow relatively slowly, but could also achieve effective denitrification and the final product was N2. In denitrification of aquaculture wastewater, strain G1 can remove 60.57% of nitrate and 36.36% of total nitrogen; meanwhile, there was a slight accumulation of ammonia nitrogen. P. denitrificans strain G1 has potential in denitrification processes for the treatment of aquaculture wastewater. However, the regulation of reaction conditions and gas products needs to be further studied.

Effects of water recirculation rate on the microbial community and water quality in relation to the growth and survival of white shrimp (Litopenaeus vannamei).

BACKGROUND: Microbial community and its management are crucial to the stabilization of culture environment for recirculating aquaculture system (RAS). Although several studies have been carried out for the microbial community of RAS, few studies were on the RAS for shrimp. Water recirculation ratio is an important factor for the microbial community and the management of RAS. Therefore, low (LC), medium (MC) and high (HC) recirculation ratio systems were set to explore the microbial community constitution of RAS for Litopenaeus vannamei and study the effect of water recirculation rate on it. RESULTS: The bacterial community of bioreactor was mainly dominated by Proteobacteria (41.6-70.7%), followed with Planctomycetes (12.5-31.0%), Bacteroidetes (10.5-26.0%), Actinobacteria (1.1-4.8%) and Verrucomicrobia (1.4-6.8%) phylum. The most dominant family of bioreactor was Rhodobacteraceae or Planctomycetaceae. The bacterial community of culture water was simpler than bioreactor and dominated by Proteobacteria (61.8-96.4%). The dominant bacterial groups of bioreactor and culture water are also different among the three water recirculation rates, and the proportions of dominant groups showed a trend with the variety of water recirculation rate. Water quality indexes including ammonia and nitrite decreased with the increasing of water recirculation rate. According to the growth performance of L. vannamei, shrimp had better performance of growth rate and final weight in MC and HC, however, shrimp had higher survival and yield in LC. Shrimp survival and yield had an inverse correlation with water recirculation rate. CONCLUSIONS: The results demonstrate the microbial community of RAS for shrimp, highlight the importance of further studies on the function of bacterial taxa, and promote the understanding of the effects of water recirculation rate on the microbiota. The findings suggest that water recirculation rate has important impacts on the microbial community, water quality and shrimp growth. Increasing the water recirculation rate could improve the water quality and promote the growth of shrimp. However, the survival rate and yield of L. vannamei are higher under low water recirculation rate. Recirculation rate is an effective method to manage RAS, and its impact on RAS needs further study, especially in the application of low level of water recirculation.

Adding maize cobs to vertical subsurface flow constructed wetlands treating marine recirculating aquaculture system effluents: Carbon releasing kinetics and intensified nitrogen removal.

This study aims to investigate the releasing behaviors of maize cobs with or without alkali (i.e. NaOH) pretreatment in seawater and distilled water, and to evaluate the effects of maize cobs addition (solid biomass and lixivium) on nitrogen removal in saline constructed wetland (CW) treating marine recirculating aquaculture system (RAS) effluents. Results revealed NaOH-treated maize cobs released carbon more efficiently, whether in seawater or in distilled water. Compared to distilled water, seawater conditions promoted carbon releasing. CW with maize cobs biomass and lixivium addition had high NO3-N removal efficiencies without significant difference, i.e. 94.9 ±â€¯6.0% and 87.1 ±â€¯13.2%, respectively. While CW with maize cobs biomass addition had higher effluent COD concentrations (16.3 ±â€¯3.6 mg L-1) compared with those adding lixivium (2.1 ±â€¯0.4 mg L-1). The study suggested adding maize cobs lixivium to be feasible and effective way to enhance nitrogen removal in saline CW.

Nutrients removal and substrate enzyme activities in vertical subsurface flow constructed wetlands for mariculture wastewater treatment: Effects of ammonia nitrogen loading rates and salinity levels.

This study aims to investigate the effects of ammonia nitrogen loading rates and salinity levels on nutrients removal rates and substrate enzyme activities of constructed wetland (CW) microcosms planted with Salicornia bigelovii treating mariculture wastewater. Activities of urease (UA), dehydrogenase (DA), protease (PrA) and phosphatase (PA) were considered. Using principal component analysis (PCA), nutrient removal index (NRI) and enzyme activity index (EAI) were developed to evaluate the effects. The results revealed that increasing ammonia nitrogen loading rates had positive effects on nitrogen removal rates (i.e. NH4-N and DIN) and enhanced substrate enzyme activities. Compared with low salinity (i.e. 15 and 22), high salinity levels (i.e. 29 and 36) enhanced nutrients removal rates, DA and UA, but weaken PA and PrA. In conclusion, CW microcosms with Salicornia bigelovii can be used for the removal of nutrients under a range of ammonia nitrogen loadings and high salinity levels.

Ammonia-oxidizing bacteria and archaea within biofilters of a commercial recirculating marine aquaculture system.

While biofilters are widely used to metabolize ammonia and other wastes in marine recirculating aquaculture systems, the ammonia-oxidizing bacterial and archaeal communities have not been characterized across a diversity of production systems. Using a metagenomics approach, we characterized the ammonia-oxidizing microbiological community of biofilters in a commercial recirculating marine aquaculture system producing hybrid grouper (Epinephelus lanceolatus × E. fuscoguttatus). Cloning and sequencing of the amoA gene showed that nitrifying bacteria included Nitrosomonas europea, N. stercoris, N. cryotolerans, N. eutropha, N. estuarii, eight strains of N. marina, and 15 strains not associated with described species. Nitrifying archaea included eight strains of Nitrosopumilus maritimus, N. koreensis, N. piranensis, N. adriaticus, undescribed congeners, and other undescribed archaea. The species composition of the bacterial and especially the archaeal communities was beyond that yet reported for aquaculture biofilters. While ammonia flux through the respective communities has yet to be estimated, the diverse environmental adaptations of the bacterial and archaeal communities suggest resilience of function under a range of environmental conditions.