Fish Aquaculture Research Articles

Pichia pastoris composition expressed aerolysin mutant of Aeromonas veronii as an oral vaccine evaluated in zebrafish (Danio rerio)

Vaccines are one of the most practical means to stop the spreading of Aeromonas veronii in aquaculture. In this study, virulence factor aerolysin mutant NTaer which has lost its hemolytic activity was used as a target antigen. Pichia pastoris constitutive secretory expression NTaer (GS115-NTaer) was used as a potential safe oral vaccine to evaluate its effectiveness on zebrafish immunity. The result shows that vaccination of GS115- NTaer for four weeks did not affect the growth performance of the host, while eliciting an effective immune protective response. Compared with the control group, the GS115-NTaer could significantly up-regulate the relative expression level of the intestinal tight junction protein 1α (TJP1α) gene, and significantly increased the contents of lysozyme (LYZ), complement C3 and C4 in the gut, indicating that the innate immune response of the fish was activated. The relative gene expression levels of macrophage-expressed gene 1 (MPEG1) and T cell receptor (TCR-α) in the gut, and MPEG1, CD4, CD8, TCR-α, GATA3, and T-bet in the spleen were all increased significantly, indicating that the cellular immune response of the fish was activated. Furthermore, the contents of serum IgM and intestinal mucosa IgZ antibodies were significantly increased, which showed that humoral immunity was also activated. Moreover, inoculation with GS115-NTaer significantly changed the structure of gut microbiota. In particular, the relative ratio of (Firmicutes + Fusobacteriota + Bacteroidota)/Proteobacteria was significantly higher than that of the control and GS115 groups. Lastly, the vaccinated fish were challenged with A. veronii, and the relative percent survival of GS115 and the GS115-NTear groups was 14.28% and 33.43%. This improvement of immunity was not only due to the specific immune response but also attributed to the improvement of innate immunity and the gut microbiota which was demonstrated by the germ-free zebrafish model. Collectively, this study provides information on the effectiveness of GS115-NTear as an oral vaccine for the green prevention and control of A. veronii infection in fish aquaculture.

Sea urchin waste as a promising nutritional source for the polychaete Hediste diversicolor (OF Müller, 1776)

In aquaculture, species diversification emerges as an imperative sustainable strategy, essential for mitigating both biological and financial risks. In this context, the cultivation of sea urchins has gained prominence as an emerging trend. However, its commercial viability still faces several challenges, namely limited growth performance. Integrated multi-trophic aquaculture (IMTA) represents a potentially more profitable model by integrating extractive species to mitigate ecological impacts and promote circular economy principles. The use of polychaetes in IMTA has proven effective in recycling waste from fish aquaculture production, however, their potential should be further explored in low-trophic aquaculture solutions. This study evaluated the potential of reared sea urchin biodeposits as a nutritional feed source for the polychaete Hediste diversicolor (OF Müller, 1776). H. diversicolor juveniles were fed three diets, over 50 days: sea urchin waste (SUW), fish waste (FW) and fish feed (FF). Survival, growth, and protein and fatty acid (FA) contents were assessed. Total survival was observed in all treatments except for FW (98%). Polychaetes fed SUW and FF presented similar specific growth rates. The protein level in the polychaetes fed SUW was significantly lower, reflecting the diet's composition. The FA profile of H. diversicolor was also influenced by the respective diets. The absolute content (μg mg−1 DW) and FA profile of polychaetes were significantly different among treatments, reflecting the composition of the diets. Saturated FA (SFA) represented the dominant class in the polychaetes fed with waste diets, while in FF polychaetes polyunsaturated fatty acids (PUFA) were the major group. Nevertheless, polychaetes fed the waste diets demonstrated the PUFA biosynthetic capacity of this species, showing a considerably higher proportion of essential long-chain FA, compared to diets. Overall, these preliminary results suggest, for the first time, the potential for recycling sea urchin biodeposits into a polychaete biomass with high commercial value, presenting a promising opportunity for low-trophic integrated aquaculture models.

Effect of Pond-Based Rice Floating Bed on the Microbial Community Structure and Quality of Water in Pond of Mandarin Fish Fed Using Artificial Diet.

The culture of mandarin fish using artificial feed has been gaining increasing attention in China. Ensuring good water quality in the ponds is crucial for successful aquaculture. Recently, the trial of pond-based rice floating beds (PRFBs) in aquaculture ponds has shown promising results. This research assessed the impact of PRFBs on the microbial community structure and overall quality of the aquaculture pond, thereby enhancing our understanding of its functions. The results revealed that the PRFB group exhibited lower levels of NH4+-N, NO2--N, NO3--N, TN, TP, and Alk in pond water compared to the control group. The microbial diversity indices in the PRFB group showed a declining trend, while these indices were increasing in the control group. At the phylum level, there was a considerable increase in Proteobacteria abundance in the PRFB group throughout the culture period, suggesting that PRFBs may promote the proliferation of Proteobacteria. In the PRFB group, there was a remarkable decrease in bacterial populations related to carbon, nitrogen, and phosphorus metabolism, including genera Rhodobacter, Rhizorhapis, Dinghuibacter, Candidatus Aquiluna, and Chryseomicrobium as well as the CL500_29_marine_group. Overall, the research findings will provide a basis for the application of aquaculture of mandarin fish fed an artificial diet and rice floating beds.

Feed Components and Timing to Improve the Feed Conversion Ratio for Sustainable Aquaculture Using Starch.

Aquaculture contributes to the sustainable development of food security, marine resource conservation, and economy. Shifting aquaculture feed from fish meal and oil to terrestrial plant derivatives may result in cost savings. However, many carnivorous fish cannot be sustained on plant-derived materials, necessitating the need for the identification of important factors for farmed fish growth and the identification of whether components derived from terrestrial plants can be used in feed. Herein, we focused on the carnivorous fish leopard coral grouper (P. leopardus) to identify the essential growth factors and clarify their intake timing from feeds. Furthermore, we evaluated the functionality of starch, which are easily produced by terrestrial plants. Results reveal that carbohydrates, which are not considered essential for carnivorous fish, can be introduced as a major part of an artificial diet. The development of artificial feed using starch offers the possibility of increasing the growth of carnivorous fish in aquaculture.

Tracking Biomarkers for the Health and Welfare of Aquaculture Fish

Aquaculture production has been growing consistently over the last few decades to meet the increasing animal protein demand of the human population. However, increased production and rearing intensities raise the challenges of guaranteeing fish health and welfare, which is essential to avoid losses and ensure product quality. Biomarkers can provide insights into the fish’s nutritional, physiological, and health status, and aid in the evaluation of early nutritional and physiological imbalances, distress conditions, and pathological diagnosis. The discovery and validation of biomarkers rely mostly on the use of information provided by different parameters, including biochemical, metabolic, or immunologic, as well as several omics, from genomics and transcriptomics to proteomics and metabolomics. In this review, a summary of the main biomarkers used in aquaculture is provided along with an overview of the main omics technologies available for further biomarker research. This review also highlights the need to develop non-lethal biomarkers that can easily and quickly be measured to provide a prompt response to producers.

Hydrolysis Optimization of Thraustochytrid (Heterotrophic Protist) Biomass for the Production of a New and Sustainable Ingredient for the Aquafeed Industry

Thraustochytrids biomass, rich in docosahexaenoic acid (DHA), could be a sustainable ingredient for the aquafeed industry. However, its production cost renders it less competitive than fishmeal and fish oil from wild-caught fish. This study proposes optimizing the hydrolysis conditions of thraustochytrid biomass to generate an ingredient with improved properties thanks to the production of peptides with different biological activities. The improved nutritional value of the biomass could justify its use to decrease the amount of wild-caught fish in aquafeed, which would enhance the sustainability of fish aquaculture. First, two commercial proteases, Papain and Alcalase 2.4L, were compared for their capacity to hydrolyze the biomass. The best degree of hydrolysis (19 ± 1%) was obtained with Alcalase 2.4L, which was then used for the optimization of the pH, enzyme concentration and reaction time using response surface methodology. The results showed that the highest concentration of peptides and DHA in the aqueous phase was obtained with a pH, enzyme concentration and reaction time of 7.5, 2.7% and 205 min, respectively. If thraustochytrid hydrolysates prove to improve fish fitness, this mild and free-solvent process protocol could be used to produce a sustainable ingredient for aquafeed.

C-Jun N-terminal kinase 1/P53 signaling mediates intrinsic apoptosis of largemouth bass (Micropterus salmoides) hepatocytes under heat stress

The increasing frequency of high-temperature extremes threatens largemouth bass Micropterus salmoides, a significant fish for freshwater ecosystems and aquaculture. Our previous studies at the transcript level suggested that heat stress induces hepatic apoptosis in largemouth bass. In the current study, we sought to validate these findings and further investigate the role of the c-Jun N-terminal kinase (JNK)/P53 signaling in hepatic apoptosis under heat stress. First, heat treatments were conducted in vivo and in vitro under different temperatures: 28 °C, 32 °C, and 37 °C. In primary hepatocytes subjected to heat treatment, cell viability was evaluated via the Cell Counting Kit-8, while mitochondrial membrane potential and nuclear morphology were assessed through JC-1 and Hoechst 33258 staining, respectively. We observed reductions in both cell viability and mitochondrial membrane potential (ΔΨm), along with alterations in nuclear morphology, in primary hepatocytes exposed to heat stress at temperatures of 32 °C and 37 °C. Quantitative real-time PCR revealed significant alterations in the expression profiles of intrinsic apoptosis-related genes within liver tissues under heat stress. Immunohistochemistry analysis revealed that JNK1 signaling increased as the temperature increased, JNK2 expression increased only at 37 °C, and JNK3 expression did not change with temperature. We speculate that JNK1 and JNK2 have pro- and anti-apoptotic effects, respectively. Western blot analysis conducted on cultured hepatocytes further validated these findings. JNK inhibition reduced hepatocyte apoptosis, improved nuclear morphology, and maintained ΔΨm even after 37 °C treatment. These results not only confirm that heat stress led to intrinsic apoptosis of hepatocytes but also indicated that JNK1 could mediate P53 expression and activate caspase-dependent intrinsic apoptosis in largemouth bass hepatocytes under such conditions. This study illuminates the physiological responses of largemouth bass to acute heat stress, offering valuable insights into the potential impacts of climate change on freshwater fishes and the sustainability of aquaculture.

Research on Sea Trial Techniques for Motion Responses of HDPE Floating Rafts Used in Aquaculture

The innovative aquaculture equipment known as high-density polyethylene (HDPE) floating rafts has gained popularity among fishermen in the southeast coastal regions of China. Compared to deep-water anti-wave fish cages, the construction costs of HDPE floating rafts are 50% to 75% less. There is a dearth of comprehensive publicly available records of HDPE floating rafts sea trial data, despite substantial numerical studies on the motion response of aquaculture fish cages and scale model experiments under controlled-wave conditions. This study involves sea trial techniques under operational and extreme environmental conditions for motion responses of HDPE floating rafts, presents a comprehensive procedure for sea trials of HDPE floating rafts, summarizes the issues encountered during the trials, and suggests solutions. Using MATLAB for independent programming, motion videos and photos collected from the sea trials are processed for image capture, yielding the original time history curve of vertical displacement. Based on the sea trials’ data, including motion displacement, acceleration, mooring line force, overall deformation patterns, and current and wave data, recommendations are provided for the design and layout of HDPE floating rafts. Based on the Fast Fourier Transform (FFT) method for spectral analysis, the influence of interference items on the observational data is eliminated; the rationality of the observational data is verified in conjunction with the results of the Gabor Transform. This study offers a scientific analytical method for the structural design and safe operation of HDPE floating rafts and provides a reference for subsequent numerical simulations.

Navigating aquaculture losses: Tackling fish‐killing phytoflagellates in a changing global landscape

AbstractAquaculture has emerged as a pivotal alternative for seafood production, addressing the challenges posed by depleting wild fish stocks. However, the industry faces substantial challenges, notably the presence of harmful algal blooms (HABs), which can cause serious economic consequences. The proliferation of major species of phytoflagellates leading to massive fish mortality is on the rise, possibly attributed to increasing global temperatures and eutrophication. This underscores the critical need for robust detection, prevention, and mitigation strategies to safeguard the aquaculture industry. This contribution offers valuable insights into comprehending and addressing the challenges posed by HABs in the fish aquaculture sector in an era of global change.

More than just geosmin and 2‐methylisoborneol? Off‐flavours associated with recirculating aquaculture systems

AbstractThe consumption of seafood is driven by flavour, yet achieving its high quality remains a challenge for many species reared in recirculating aquaculture systems (RAS). A comprehensive knowledge of off‐odour sources in aquatic foods is indispensable in ensuring flavour quality standards. At the beginning of the production chain, early post‐harvest lipid oxidation products develop into endogenous off‐odours and accumulate over time. These malodours add to those already absorbed exogenously, namely from the water and feeds, although the information on the interactions between these sources is currently scarce. Despite geosmin and 2‐methylisoborneol receiving significant attention in relation to fish off‐flavour, only limited knowledge on the molecular and ecological mechanisms driving their production in aquatic environments has been reported. Moreover, RAS‐hosted bacteria have been associated with a wide range of other odour‐active compounds, such as pyrazines, terpenoids, and other degradation byproducts that are frequently overlooked when studying flavour taint in fish. The influence of aquaculture feeds on the flavour of fish flesh has been underestimated, too, both as a source of off‐odours but also as a novel modulator strategy to achieve desirable aquatic food flavours. Finally, the influence of water treatment processes widely used in RAS operations, such as advance oxidation process, ozone, ultraviolet and hydrogen peroxide disinfections, is greatly underexplored with respect to odour quality. This article reviews the current scientific evidence with supporting data on the chemical diversity of off‐odours found in aquaculture fish worldwide and their potential sources and highlights knowledge gaps that should be addressed in future research.

Effects of dietary tryptophan levels on growth performance, serotonin metabolism, brain 5-HT and cannibalism activities in tiger puffer, Takifugu rubripes fingerlings

Essential amino acid (EAA) requirements of aquaculture fish species can be used as a guideline for commercial feed formulation. Several EAA requirements have been quantified in tiger puffer (Takifugu rubripes). However, as the third limiting amino acid (AA) of many plant protein sources, tryptophan requirement of this species has yet to be quantified. This study was conducted to determine dietary tryptophan requirement of tiger puffer, and investigate the effect of dietary tryptophan supplementation on the cannibalism, brain serotonin (5-hydroxytryptpamine, 5-HT) metabolism and serum cortisol levels. Six experimental diets were prepared using fish meal, gelatin and corn protein concentrate as intact protein, and then supplemented with crystalline AA premix. The analyzed tryptophan levels in the experimental diets were 3.5, 4.1, 4.5, 5.5, 7.0 and 11.8 g/kg of dry diet. Experimental diets were randomly fed in triplicate to tiger puffer (initial average weight of 17.5 g) for 58 days. The weight gain (WG), feed conversion ratio (FCR) and protein efficiency ratio (PER) were significantly affected by dietary tryptophan, and the break points were observed at 5.07, 4.82 and 4.99 g/kg of dry diet, respectively. Compared to the basal diet, the cannibalism (the rate of bite mark appearance) was significantly lower and survival rate was significantly higher when fish were given diets containing 7.0 and 11.8 g/kg tryptophan. The concentrations of free tryptophan and phenylalanine in the brain exhibited a nonlinear increase in response to the incremental levels of dietary tryptophan. Brain 5-HT concentration was significantly affected by dietary tryptophan levels, but did not show a significant dose-dependent response. The expression of genes related to 5-HT metabolism (tph1a, ddc and aldh) and serotonin reuptake (sert) in the brain varied significantly and exhibited a nonlinear relationship with the graded levels of dietary tryptophan. In addition, serum cortisol levels showed a decreasing trend till the 5.5 g/kg tryptophan inclusion level, at which point there appeared to be the lowest level. In conclusion, based on nonlinear regression analysis of the WG, FCR and PER, dietary tryptophan requirement of tiger puffer was determined to be range between 4.82 and 5.07 g/kg of dry diet (10.48–11.03 g/kg of dietary protein). Dietary tryptophan supplementation above requirement could mitigate cannibalism and improve the survival of tiger puffer, which may be attributed to regulating the synthesis and catabolism of 5-HT and its availability in the synaptic cleft, and subsequently the reduction of serum cortisol content.

The effect of an organomineral chelate complex on the amylolytic activity of intestinal enzymes of some fish species

To date, there is a large amount of information regarding the effect of heavy metals on the hydrolases of the digestive tract. It has been established that even biogenic metals in high concentrations can have a toxic effect. The assessment of the activity of digestive hydrolases is relevant, since a change in their activity has a direct effect on the effectiveness of assimilation of food. Amylolytic hydrolases help to provide the body with carbohydrates (along with fats) – the main sources of energy for fish. A decrease in the intake of carbohydrates will lead to a violation of metabolic processes and, as a result, underdevelopment and death of fish. The aim of the study was to determine the effect of an organomineral chelate complex on the amylolytic activity of intestinal enzymes of some fish species. The research was conducted at the Department of Biochemistry and Physiology of the St. Petersburg State University of Veterinary Medicine and on the basis of two fisheries in the Leningrad Region. The fish of the experimental groups (n = 10) were given an additive for 30 days, the data were compared with the control (n = 10). The drug was given together with the feed once a day, at a concentration of 0.5 g / kg (feed weight). Fingerlings (0+) of 3 species of freshwater bony fish were used as research objects: Carassius auratus (n=20), Cyprinus carpio (n=20), Oncorhynchus mykiss (n=20). Amylolytic activity was determined in the total homogenets of the intestinal mucosa of fish. The activity was assessed by the increase in hexoses by the Nelson method in the modification of Ugolev, Jesuitova. The detection was carried out on a photoelectric KKK-3 photometer. Based on the studies obtained, it can be concluded that the use of an organomineral chelate complex in the studied dosage (previously worked out in conditions of determining acute and chronic toxicity for fish) does not have a negative effect on the amylolytic activity of intestinal enzymes. There was no significant decrease in amylolytic activity in all experimental groups. Based on this, it is possible to recommend the use of a chelate complex for fish in industrial aquaculture.

Sterilization of fish through adaptable gRNAs targeting dnd1 using CRISPR-Cas13d system

Sterilizing fish is a valuable technique not only for preventing genetic contamination from escaped aquaculture fish but also for enhancing fish meat quality. The clustered regularly interspaced short palindromic repeats-CRISPR-associated 13d (CRISPR-Cas13d) system is capable of RNA-targeted gene knockdown with reduced off-target effects. Unlike morpholino knockdown, which is limited to specific target sites, the CRISPR-Cas13d system utilizes the entire mRNA as a target site for gRNA. This enables the design of adaptable gRNAs by targeting the conserved sequences across fish species, leading to promising sterilization for a wide variety of aquaculture fish. In this study, we demonstrated that the effectiveness of this system in fish sterilization by the knockdown of dead end1 (dnd1) and nanos3, which are crucial for germ cell development. Initially, we demonstrated that gRNAs designed using the cas13design tool, which was developed based on studies of cultured mammalian cells, effectively induced germ cell-deficient medaka with low-cytotoxicity. Additionally, we identified the nucleotide positions of the gRNA (seed sequence) that are crucial for knockdown in teleost fish. These data will be valuable for designing adaptable gRNA in teleost fish and targeting multiple paralogous genes. To develop an adaptable sterilization technique using the CRISPR-Cas13d system, we aligned 81 dnd1 sequences from 70 fish species and identified a highly conserved region of the gRNA targets. Despite not finding a perfectly matched target region in the dnd1 alignment, we designed 15 adaptable gRNAs that potentially targeting dnd1 in 55 out of 70 fish species, leveraging sequences from representative aquaculture and aquarium fish (salmon, tilapia, puffer fish, amberjack, tuna, guppy and carp) and experimental model fish (medaka, zebrafish, and stickleback. To confirm whether adaptable gRNAs had a knockdown effect, we initially injected the gRNAs into medaka embryos. Adaptable gRNAs with a single-nucleotide mismatch against medaka dnd1 resulted in the complete loss of germ cells with little cytotoxicity, suggesting that mismatches are unlikely to induce off-target effects crucial for embryonic development. Additionally, adaptable gRNAs designed based on the salmon dnd1 sequence, which was 100% identical in the Oncorhynchus, Salvelinus, and Salmo groups, effectively generated germ cell-deficient rainbow trout. These results indicate that the knockdown of dnd1 using the CRISPR-Cas13d system is effective for sterilizing both aquaculture and experimental model fish species. Moreover, the adaptable gRNAs designed in this study can potentially sterilize a diverse range of fish species.

Effect of bile salts on intestinal epithelial function in gilthead seabream (Sparus aurata)

In the context of modern aquaculture, the effort to reduce the reliance on fishmeal/marine ingredients in fish diets has led to the exploration of plant-based protein sources as potential substitutes, a dietary shift that disrupts the bile acid profile in fish. Therefore, bile salts are being sought as additives. However, artificially increased intestinal levels of bile acids may significantly impact mucosal function. Therefore, here, we explored the regulatory role in the intestine of gilthead sea bream (Sparus aurata) of (i) chenodeoxycholic acid (CDC), (ii) a mixture formed by two bile acids, 3% cholic acid and 97% deoxycholic acid (MIX), and (iii) a conjugated bile salt sodium taurocholate (TC) in Ussing chambers with the epithelial voltage clamp technique. We tested the bile salts in a 50–500 μg/ml concentration range, and all of them promoted ion absorption. Yet, clear concentration-dependent and more pronounced effects on the ion transport were observed in the posterior intestine. On the other hand, bile salts had no or minor effects on tissue resistance. However, there are indications that the MIX could have adverse effects at high concentrations (500 μg/ml), promoting a threefold increase in tissue permeability measured using FITC-dextran (4 kD) regardless of the intestinal region, thus suggesting an alteration in intestinal permeability at high bile salt concentrations. The findings from our study emphasize the importance of considering intestinal function when contemplating the possible use of a particular bile salt as a dietary supplement. It appears that bile salts, whether acting individually or in combination, play a pivotal role in orchestrating nutrient absorption by influencing the function of epithelial ion transport. However further research is needed to fully grasp the region-dependent nuances of bile salt effects on ion transport and the ultimate consequences for nutrient absorption in the context of fish aquaculture.

Evolution and Prospects of Freshwater Fish Species and Aquaculture in Colombia: A Historical Perspective

Since the 1970s Colombia has been aware of the importance of aquaculture and inland fisheries in the future of its economy due to the constant increase of a population that is increasingly in need of protein of animal origin. In 1972, a joint project between the Government of Colombia and the Food and Agriculture Organization of the United Nations was approved and aimed at studying inland fishing and developing aquaculture in the country. The work of the project is described, as are the fish culture and fisheries research activities starting with the situation in the 1970s but comparing it with recent findings. A list of the main capture and culture fish species is given. Colombian fishing production has had great variations, but first inland water fishing was more important than marine. For instance, in 1972 total fish catch was 104,390 tons and 79% of that originated from inland waters. After the continental fisheries went down, the marine fisheries accounted in 2019 already 78 per cent of the total catch of the country. Mean standing fish crop estimates in 1975‒76 from the bay, open water and vegetation habitats in four floodplain lakes were 118, 12 and 251 kg/ha indicating clearly that the fish productivity on the Magdalena floodplain was relatively low. The aquaculture sector has been growing rapidly, reflecting global trends, and producing more than capture fisheries. Aquaculture produced 88,000 tons in 2013, which was a three-fold increase from the mid-1990s. Again twofold increase took place before 2020 when the production was almost 180,000 tons. Future development prospects of inland fisheries and aquaculture are discussed and a mention is made of the present situation of ornamental fish cultivation and trade in Colombia.

Bee venom enhances performance and immune function in thinlip mullet: A promising approach for sustainable aquaculture

As an environmentally friendly alternative to antibiotics, bee venom holds promise for aquaculture due to its diverse health advantages, including immune-amplifying and anti-inflammatory features. This study investigated the effects of dietary bee venom (BV) on the growth and physiological performance of Thinlip mullet (Liza ramada) with an initial body weight of 40.04 ± 0.11 g for 60 days. Fish were distributed to five dietary treatments (0, 2, 4, 6, and 8 mg BV/kg diet) with three replicates. Growth traits, gut enzyme ability (lipase, protease, amylase), intestinal and liver histology, blood biochemistry, immune responses [lysozyme activity (LYZ), bactericidal activity (BA), nitroblue tetrazolium (NBT%)], and antioxidant status [superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), malondialdehyde (MDA)] were evaluated. BV supplementation significantly improved growth performance, digestive enzyme activity, histological integrity of organs, immune responses (LYZ, BA), and antioxidant status (SOD, CAT, GPx), while declining MDA levels. Optimal BV levels were identified between 4.2 and 5.8 mg/kg diet for different parameters. Overall, the findings suggest that BV supplementation can enhance growth and physiological performance in Thinlip mullet, highlighting its potential as a beneficial dietary supplement for fish health and aquaculture management.

Knockout of EPO gene in blunt snout bream (Megalobrama amblycephala) by CRISPR/Cas9 reveals its roles in hypoxia-tolerance

Erythropoietin (EPO) is a glycoprotein hormone that plays a crucial role as a downstream regulator in the Hypoxia-inducible Factor (HIF) signaling pathway, mainly involved in the regulation of erythropoiesis. The open reading frame (ORF) of EPO in blunt snout bream (Megalobrama amblycephala, BSB) was 561 bp, encoding 186 amino acids. Phylogenetic analysis revealed it clusters with homologs in others teleost. We designed two target sites (targets 1, 2) to obtain gene knockout mutants, with the mature mRNA sequence of EPO that were located in exon 2, exon 3, respectively. The mutation efficiency of targets 1, 2 was 19.60%, 24.22%, respectively, both containing 6 mutation types. F1EPO+/− mutants with the same mutation type were screened, and select F2EPO−/− mutants after mating. At the age of five months, EPO−/− fish demonstrated a significant (p < 0.05) increase in the average oxygen tension threshold for the loss of equilibrium (LOEcrit) and the shedding degree of interlamellar cell masses (ILCM), whereas a significant (p < 0.05) decrease was observed in the number of red blood cells (RBC) and the concentration of hemoglobin (Hb) under hypoxia conditions, in comparison to the control group. Notably, injection of EPO mRNA rescued the downregulation of EPO expression in EPO−/− mutant, and overexpression of the EPO gene did not disturb the normal development of BSB embryos. These findings provide new insights into the potential for applying CRISPR/Cas9 in other aquaculture fish species and the breeding of hypoxia-tolerant BSB.

Probiotic bacteria as a healthy alternative for fish and biological control agents in aquaculture

Probiotic bacteria as a healthy alternative for fish and biological control agents in aquaculture

Reducing baseline toxicity in fishery product-related sediments from land to sea: Region-specific solutions are required

Eastern China is a major producer of fishery products (including inland aquaculture, coastal mariculture, and coastal fishing products). The quality of the products is affected by hydrophobic organic contaminants (HOCs) in the sediments. Based on in-vitro luminescent bacterial assay, the baseline toxicity (BEQBio) of 56 common HOCs were assessed in the present study. Specifically, the BEQBio of sediments declined from land (31–400 mg/kg) to sea (9.1–270 mg/kg). However, the toxicity contribution explained by the HOCs increased gradually from land (0.70 %) to sea (10 %) using Iceberg Modeling. In the inland pond, current use HOCs (pyrethroid pesticide (PEs), organic tin (OTCs), and antibiotic) exhibited considerable concentrations, although their toxicity contribution was very small (0.076 %), thus more regulations on the use of HOCs should be proposed and further screening is needed to confirm the major toxicants. In coastal mariculture area, the toxicity contribution of current use HOCs further declined (0.010 %), whereas environmental background HOCs, such as polycyclic aromatic hydrocarbons (PAHs), became increasingly significant, with the contribution ratio increasing from 0.37 % to 2.4 %. To minimize the negative impacts of PAHs, optimization of energy structure in transportation and coastal industry is required. In the coastal fishing area, the phased-out persistent organic pollutants (POPs) remained a major concern, in terms of both concentration and toxicity contribution. The phased-out POPs explained 7.0 % of the toxic effects of the sediments from the coastal fishing area, due to historical residue, industrial emissions, and their high toxicities. For this reason, it is critical to improve the relevant emission regulations and standards, so as to eventually reduce the unintentional discharges of POPs.

Rice-fish symbiosis improves the muscle nutrition and intestinal flora diversity of tilapia

Rice-fish symbiosis is an ecological farming model that can improve fish’s muscle nutrient profile and intestinal flora. We investigated the effect of rice-fish symbiosis on tilapia’s intestinal flora and muscle nutrients. We used 16S rDNA technology to analyze the intestinal flora of tilapia in pond culture and rice-fish symbiosis. Furthermore, spearman correlation analysis was performed on tilapia intestinal flora and muscle nutrients. The results showed that the contents of Valine, Phenylalanine, and Docosahexaenoic acid were significantly higher in the rice-fish symbiosis than those in the pond culture (p&lt;0.05). Intestinal flora analysis showed that the diversity in rice-fish symbiosis was significantly higher than in pond culture (p&lt;0.05). Meanwhile, the abundance of Brevundimonas, Pseudomonas, and Ralstonia in rice-fish symbiosis was significantly higher than those in pond culture (p&lt;0.05), but Pirellulaceae unclassified and Cyanobium_PCC-6307 were opposite. The number of 8 predictive functional tilapia genes in the rice-fish symbiosis was higher than in the pond culture. Correlation analysis showed that the abundance of Ralstonia was positively correlated with the contents of Valine, Phenylalanine, and Docosahexaenoic acid, respectively. Our study shows that the rice-fish symbiosis could improve muscle nutrient profile and intestinal flora diversity in tilapia. This study will help to improve the rice-fish symbiosis model and provide a scientific basis for ecological aquaculture and healthy fish production.