Recirculating aquaculture systems (RASs) offer significant advantages in aquaculture by markedly decreasing water usage and increasing culture density. A vital component within a RAS is the filler material, which serves as a surface for microbial colonization. Effective microbial treatment is crucial for the efficient operation of a RAS as it assists in purifying the wastewater generated within the system. Nevertheless, traditional fillers often show low efficiency in biofilm formation. The commercial silicon carbide used in this study is a foam ceramic filter with a density of about 0.4-0.55 g/cm3, a number of holes of about 10, and a through porosity of 80.9%, with a diameter of about 5 cm. This research investigates the utilization of a titanium dioxide-silicon carbide (TiO2-SiC) composite filler to improve the purification efficiency of ammonia nitrogen and chemical oxygen demand (COD) in aquaculture wastewater. The study involved the application of titanium dioxide films onto the surface of silicon carbide to produce the composite filler. This method takes advantage of the dipole interaction between titanium dioxide and microorganisms, which enhances biofilm culturing efficiency on the silicon carbide surface. The performance of three different fillers was assessed for their ability to purify aquaculture wastewater. Results showed that the TiO2-SiC composite filler was 1.67 times more effective in removing COD and 1.07 times more effective in removing ammonia nitrogen compared to using silicon carbide alone. These results demonstrate that the incorporation of a titanium dioxide coating substantially boosts the microbial colonization efficiency of silicon carbide, thereby enhancing the overall wastewater purification efficiency in RAS.

Monopterus albus is one of China\'s renowned and superior aquaculture species, with its seedlings mainly sourced from wild capture. One of the bottlenecks in M. albus aquaculture is the high mortality rate and low feeding initiation rate from stocking wild fry to the initiation of feeding. In production, trash fish is commonly used to wean M. albus juveniles onto feeding. In this study, we introduced three other natural feeds, earthworms (EW), yellow mealworms (YMW), and fly maggots (FM), with frozen trash fish (TF) serving as the control group, to evaluate the effects of these four natural feeds on the survival rate, feeding initiation, antioxidant enzymes activity, and body composition of M. albus juveniles under recirculating water aquaculture conditions. The experiment comprised four treatments, each with three replicates. Each replicate consisted of stocking 150 M. albus juveniles weighing 10.02 ± 0.89 g in size, raised for 5 weeks. The survival rate of the YMW group was 73.33%-85.33%, which was significantly higher than that of the other three bait groups (p < 0.05). The four bait groups showed no significant differences in final body weight and specific growth rate (SGR) (p > 0.05). The EW group showed the highest final body weight, with an average SGR of 2.73, whereas the YMW group had an average SGR of 1.87. The average daily feeding amount was significantly higher in EW and YMW groups than in the other two groups (p < 0.05). The percentage of feeding amount to fish weight in the EW group reached 7.3% in the fifth week. After 5 weeks of cultivation, NO2 --N content was significantly higher in the waters of the TF and EW groups than in the waters of the FM and YMW groups (p < 0.05), there was no significant difference in TAN content among the treatment groups (p > 0.05). Liver malondialdehyde content was significantly higher in the TF group than in the other bait groups (p < 0.05). GSH-Px activity was significantly higher in the EW group than in the FM group and YMW group. No significant differences in SOD and CAT activity and T-AOC were observed among the bait groups (p > 0.05). The increase in crude protein content was significantly higher in the TF group than in the FM group, but the increase in crude ash content was significantly lower in the TFgroup. In conclusion, Tenebrio molitor could potentially serve as one of the alternative feeds during the initial stages of M. albus juveniles stocking.

Hydrogen sulphide (H2S) is considered an immunotoxicant, and its presence in the water can influence the mucosal barrier functions of fish. However, there is a significant knowledge gap on how fish mucosa responds to low environmental H2S levels. The present study investigated the consequences of prolonged exposure to sub-lethal levels of H2S on the mucosal defences of Atlantic salmon (Salmo salar). Fish were continuously exposed to two levels of H2S (low: 0.05 µM; and high: 0.12 µM) for 12 days. Unexposed fish served as control. Molecular and histological profiling focused on the changes in the skin, gills and olfactory rosette. In addition, metabolomics and proteomics were performed on the skin and gill mucus. The gene expression profile indicated that the gills and olfactory rosette were more sensitive to H2S than the skin. The olfactory rosette showed a dose-dependent response, but not the gills. Genes related to stress responses were triggered at mucosal sites by H2S. Moreover, H2S elicited strong inflammatory responses, particularly in the gills. All mucosal organs demonstrated the key molecular repertoire for sulphide detoxification, but their temporal and spatial expression was not substantially affected by sub-lethal H2S levels. Mucosal barrier integrity was not considerably affected by H2S. Mucus metabolomes of the skin and gills were unaffected, but a matrix-dependent response was identified. Comparing the high-concentration group\'s skin and gills mucus metabolomes identified altered amino acid biosynthesis and metabolism pathways. The skin and gill mucus exhibited distinct proteomic profiles. Enrichment analysis revealed that proteins related to immunity and metabolism were affected in both mucus matrices. The present study expands our knowledge of the defence mechanisms against H2S at mucosal sites in Atlantic salmon. The findings offer insights into the health and welfare consequences of sub-lethal H2S, which can be incorporated into the risk assessment protocols in salmon land-based farms.

The diversity, composition and performance of microbial communities within constructed wetlands (CW) were markedly influenced by spatio-temporal variations. A pilot-scale integrated vertical-flow constructed wetland (IVCW) as the biological purification unit within a recirculating aquaculture system (RAS) was established and monitored in this study. The investigation aimed to elucidate the responses of community structure, co-occurrence networks, and assembly mechanisms of the microbial community to spatial and temporal changes. Spatially, all a-diversity indices and microbial networks complexity were significantly higher in the upstream pool of the IVCW than in the downstream pool. Temporally, the richness increased over time, while the evenness showed a decreasing trend. The number of nodes and edges of microbial networks increased over time. Notably, the stable pollutant removal efficiencies were observed during IVCW operations, despite a-diversity and bacterial community networks exhibited significant variations across time. Functional redundancy emerged as a likely mechanism contributing to the stability of microbial ecosystem functions. Null model and neutral model analyses revealed the dominance of deterministic processes shaping microbial communities over time, with deterministic influences being more pronounced at lower a-diversity levels. DO and inorganic nitrogen emerged as the principal environmental factor influencing microbial community dynamics. This study provides a theoretical foundation for the regulation of microbial communities and environmental factors within the context of IVCW.

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.

The structure and function of the water microbial community can change dramatically between different rearing modes. Yet investigations into the relationships between microbial community and water quality remain obscure. We provide the first evidence that rearing modes alter bacterial community and water quality in the rearing water of the mud crab (Scylla paramamosain) larvae. The juveniles in the recirculating aquaculture system (RAS) had a higher viability than those in the water exchange system (WES). RAS had the significantly lower levels of total ammonia nitrogen (TAN), NH3, NO2--N, total nitrogen (TN), total dissolved solids (TDS), and chemical oxygen demand than those of WES. The number of significantly different amplicon sequence variants between rearing modes increased as the larvae developed. NH3, TAN, TDS, NO2--N, and TN were closely related to the late alterations in water bacterial community. Both the FAPROTAX tool and quantitative PCR analysis showed enhanced nitrogen cycling functional potential of water bacterial community of RAS. Random forest analysis identified the enriched water bacteria especially heterotrophic bacteria such as Phaeodactylibacter, Tenacibaculum, and Hydrogenophaga, which were vital in removing nitrogenous compounds via simultaneous nitrification and denitrification. Notably, RAS could save 18.5 m3 of seawater relative to WES in larviculture on the scale of 2.5 m3. Together, these data indicate that RAS could function as microbial community and water quality management strategy in the larviculture of crab.

Atlantic salmon (Salmo salar) might encounter toxic hydrogen sulphide (H2S) gas during aquaculture production. Exposure to this gas can be acute or chronic, with heightened levels often linked to significant mortality rates. Despite its recognised toxicity, our understanding of the physiological implications of H2S on salmon remains limited. This report details the mucosal and systemic physiological consequences in post-smolt salmon reared in brackish water at 12 ppt after prolonged exposure to elevated H2S levels over 4 weeks. The fish were subjected to two concentrations of H2S: 1 µg/L (low group) and 5 µg/L (high group). An unexposed group at 0 µg/L served as the control. Both groups exposed to H2S exhibited incremental mortality, with cumulative mortality rates of 4.7 % and 16 % for the low and high groups, respectively. Production performance, including weight and condition factors, were reduced in the H2S-exposed groups, particularly in the high group. Mucosal response of the olfactory organ revealed higher tissue damage scores in the H2S-exposed groups, albeit only at week 4. The high group displayed pronounced features such as increased mucus cell density and oedema-like vacuoles. Transcriptome analysis of the olfactory organ unveiled that the effects of H2S were more prominent at week 4, with the high group experiencing a greater magnitude of change than the low group. Genes associated with the extracellular matrix were predominantly downregulated, while the upregulated genes primarily pertained to immune response. H2S-induced alterations in the metabolome were more substantial in plasma than skin mucus. Furthermore, the number of differentially affected circulating metabolites was higher in the low group compared to the high group. Five core pathways were significantly impacted by H2S regardless of concentration, including the phenylalanine, tyrosine, and tryptophan biosynthesis. The plasma levels of phenylalanine and tyrosine were reduced following exposure to H2S. While there was a discernible distinction in the skin mucus metabolomes among the three treatment groups, only one metabolite - 4-hydroxyproline - was significantly impacted by H2S. Furthermore, this metabolite was significantly reduced in the plasma and skin mucus of H2S-exposed fish. This study underscores that prolonged exposure to H2S, even at concentrations previously deemed sub-lethal, has discernible physiological implications that manifest across various organisational levels. Given these findings, prolonged exposure to H2S poses a welfare risk, and thus, its presence must be maintained at low levels (<1 µg/L) in salmon land-based rearing systems.

The virtual, digital counterpart of a physical object, referred as digital twin, derives from the Internet of Things (IoT), and involves real-time acquisition and processing of large data sets. A fully implemented system ultimately enables real-time and remote management, as well as the reproduction of real and forecasted scenarios. Under the emerging framework of Precision Fish Farming, which brings control-engineering principles to fish production, we set up digital twin prototypes for land-based finfish farms. The digital twin is aimed at supporting producers in optimizing feeding practices, oxygen supply and fish population management with respect to 1) fish growth performances; 2) fish welfare, and 3) environmental loads. It relies on integrated mathematical models which are fed with data from in-situ sensors and from external sources, and simulate several dynamic processes, allowing the estimation of key parameters describing the ambient environment and the fishes. A conceptual application targeted at rearing cycles of rainbow trout ( Oncorhynchus mykiss) in an operational in-land aquafarm in Italy is presented. The digital twin takes into account the disparate levels of automation and control that are found within this farm, and considerations are made on preferential directions for future developments. In spite of its potential, and not only in the aquaculture sector, the development of digital twins is still at its early stage. Furthermore, Precision Fish Farming applications in land-based systems as well as targeted at rainbow trout are novel developments.
A digital twin is set up for a rainbow trout ( Oncorhynchus mykiss) aquaculture farm located in northern Italy. The farm is equipped with large-scale basins, and are supplied by freshwater from the neighbouring Sarca river. Generally speaking, digital twins are virtual, digital representations which mirror and are connected to real objects, enabling real-time and remote management, as well as the reproduction of real or forecasted scenarios. This is a pioneer application aimed at supporting producers in optimizing feeding practices, oxygen supply and fish population management with respect to 1) fish growth performances; 2) fish welfare, and 3) environmental loads. The digital twin relies on sensors, Big Data, IoT (Internet of Things) and predictive mathematical models. In spite of its potential, and not only in the aquaculture sector, the development of digital twins is still at its early stage. Furthermore, Precision Fish Farming applications in land-based systems as well as targeted at rainbow trout are novel developments.

Recirculating aquaculture systems (RAS) pose unique challenges in microbial community management since they rely on a stable community with key target groups, both in the RAS environment and in the host (in this case, Solea senegalensis). Our goal was to determine how much of the sole microbiome is inherited from the egg stage, and how much is acquired during the remainder of the sole life cycle in an aquaculture production batch, especially regarding potentially probiotic and pathogenic groups. Our work comprises sole tissue samples from 2 days before hatching and up to 146 days after hatching (-2 to 146 DAH), encompassing the egg, larval, weaning, and pre-ongrowing stages. Total DNA was isolated from the different sole tissues, as well as from live feed introduced in the first stages, and 16S rRNA gene was sequenced (V6-V8 region) using the Illumina MiSeq platform. The output was analysed with the DADA2 pipeline, and taxonomic attribution with SILVAngs version 138.1. Using the Bray-Curtis dissimilarity index, both age and life cycle stage appeared to be drivers of bacterial community dissimilarity. To try to distinguish the inherited (present since the egg stage) from the acquired community (detected at later stages), different tissues were analysed at 49, 119 and 146 DAH (gill, intestine, fin and mucus). Only a few genera were inherited, but those that were inherited accompany the sole microbiome throughout the life cycle. Two genera of potentially probiotic bacteria (Bacillus and Enterococcus) were already present in the eggs, while others were acquired later, in particularly, forty days after live feed was introduced. The potentially pathogenic genera Tenacibaculum and Vibrio were inherited from the eggs, while Photobacterium and Mycobacterium seemed to be acquired at 49 and 119 DAH, respectively. Significant co-occurrence was found between Tenacibaculum and both Photobacterium and Vibrio. On the other hand, significantly negative correlations were detected between Vibrio and Streptococcus, Bacillus, Limosilactobacillus and Gardnerella. Our work reinforces the importance of life cycle studies, which can contribute to improve production husbandry strategies. However, we still need more information on this topic as repetition of patterns in different settings is essential to confirm our findings.

Recirculating aquaculture systems (RAS) are increasingly being used to grow fish, as intensive water reuse reduces water consumption and environmental impact. RAS use biofilters containing nitrogen-cycling microorganisms that remove ammonia from the aquaculture water. Knowledge of how RAS microbial communities relate to the fish-associated microbiome is limited, as is knowledge of fish-associated microbiota in general. Recently, nitrogen-cycling bacteria have been discovered in zebrafish and carp gills and shown to detoxify ammonia in a manner similar to the RAS biofilter. Here, we compared RAS water and biofilter microbiomes with fish-associated gut and gill microbial communities in laboratory RAS housing either zebrafish (Danio rerio) or common carp (Cyprinus carpio) using 16S rRNA gene amplicon sequencing. The phylogeny of ammonia-oxidizing bacteria in the gills and the RAS environment was investigated in more detail by phylogenetic analysis of the ammonia monooxygenase subunit A (amoA). The location from which the microbiome was sampled (RAS compartments and gills or gut) had a stronger effect on community composition than the fish species, but species-specific differences were also observed. We found that carp- and zebrafish-associated microbiomes were highly distinct from their respective RAS microbiomes, characterized by lower overall diversity and a small core microbiome consisting of taxa specifically adapted to the respective organ. The gill microbiome was also defined by a high proportion of unique taxa. Finally, we found that amoA sequences from the gills were distinct from those from the RAS biofilter and water. Our results showed that the gut and gill microbiomes of carp and zebrafish share a common and species-specific core microbiome that is distinct from the microbially-rich RAS environment.