POF-based biosensors for cortisol detection in seawater as a tool for aquaculture systems.

A surface plasmon resonance (SPR) phenomenon implemented via D-shaped polymer optical fiber (POF) is exploited to realize cortisol biosensors. In this work, two immonosensors are designed and developed for the qualitative as well as quantitative measurement of cortisol in artificial and real samples. The performances of the POF-based biosensors in cortisol recognition are achieved using different functionalization protocols to make the same antibody receptor layer over the SPR surface via cysteamine and lipoic acid, achieving a limit of detection (LOD) of 0.8 pg/mL and 0.2 pg/mL, respectively. More specifically, the use of cysteamine or lipoic acid changes the distance between the receptor layer and the SPR surface, improving the sensitivity at low concentrations of about one order of magnitude in the configuration based on lipoic acid. The LODs of both cortisol biosensors are achieved well competitively with other sensor systems but without the need for amplification or sample treatments. In order to obtain the selectivity tests, cholesterol and testosterone were used as interfering substances. Moreover, tests in simulated seawater were performed for the same cortisol concentration range achieved in buffer solution to assess the immunosensor response to the complex matrix. Finally, the developed cortisol biosensor was used in a real seawater sample to estimate the cortisol concentration value. The gold standard method has confirmed the estimated cortisol concentration value in real seawater samples. Liquid-liquid extraction was implemented to maximize the response of cortisol in liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis.

Growth and physiological response of proactive and reactive juvenile "tambaqui" (Colossoma macropomum) in a recirculating aquaculture system.

This study evaluated the growth and physiological response of proactive and reactive Colossoma macropomum juveniles in a recirculating aquaculture system (RAS). In Phase 1 of the experiment (50 days of cultivation), juveniles, weighing 2.16 ± 0.52 g, were stocked in 12 28-L tanks to test the following treatments: proactive (PT), reactive (RT) and mixed (MT) composed of reactive (MRT) and proactive (MPT) animals. In Phase 2 of the experiment (40 days of cultivation), the animals were transferred to 175-L tanks with the same treatments as Phase 1. The animals were fed twice a day with commercial diet during both phases. After Phase 1, MPT animals showed higher growth than MRT animals (P < 0.05), and higher weight gain and daily weight than PT animals (P < 0.05). After Phase 2, PT animals showed higher weight gain and daily weight gain than RT and MT animals (P < 0.05), as did MPT animals compared to PT animals. Performance for RT animals was superior (P < 0.05) to that of MRT animals. Glucose (P < 0.04) and cholesterol (P < 0.01) were higher for RT animals compared to PT animals. Cholesterol was higher for MPT animals compared to MRT animals (P < 0.01), while plasma protein was lower (P < 0.001). Glucose (P < 0.001) and cholesterol (P < 0.01) were higher for MPT animals compared to PT animals and for MRT animals compared to RT animals (glucose P < 0.02, cholesterol P < 0.01). After 90 days of cultivation, proactive animals cultivated separately presented better performance. When cultivated together, reactive animals experienced a decrease in performance and both stress coping styles showed more signs of stress.

Advancing aquaculture: Production of xenogenic catfish by transplanting blue catfish (Ictalurus furcatus) and channel catfish (I. punctatus) stem cells into white catfish (Ameiurus catus) triploid fry.

Xenogenesis has been recognized as a prospective method for producing channel catfish, Ictalurus punctatus ♀ × blue catfish, I. furcatus ♂ hybrids. The xenogenesis procedure can be achieved by transplanting undifferentiated stem cells derived from a donor fish into a sterile recipient. Xenogenesis for hybrid catfish embryo production has been accomplished using triploid channel catfish as a surrogate. However, having a surrogate species with a shorter maturation period, like white catfish (Ameiurus catus), would result in reduced feed costs, labor costs, and smaller body size requirements, making it a more suitable species for commercial applications where space is limited, and as a model species. Hence, the present study was conducted to assess the effectiveness of triploid white catfish as a surrogate species to transplant blue catfish stem cells (BSCs) and channel catfish stem cells (CSCs). Triploid white catfish fry were injected with either BSCs or CSCs labeled with PKH 26 fluorescence dye from 0 to 12 days post hatch (DPH). No significant differences in weight and length of fry were detected among BSCs and CSCs injection times (0 to 12 DPH) when fry were sampled at 45 and 90 DPH (P > 0.05). The highest survival was reported when fry were injected between 4.0 to 5.5 DPH (≥ 81.2%). At 45 and 90 DPH, cell and cluster area increased for recipients injected from 0 to 5.2 DPH, and the highest cluster area values were reported between 4.0 to 5.2 DPH. Thereafter, fluorescent cell and cluster area in the host declined with no further decrease after 10 DPH. At 45 DPH, the highest percentage of xenogens were detected when fry were injected with BSCs between 4.0 to 5.0 and CSCs between 3.0 to 5.0 DPH. At 90 DPH, the highest number of xenogens were detected from 4.0 to 6.0 DPH when injected with either BSCs or CSCs. The current study demonstrated the suitability of white catfish as a surrogate species when BSCs and CSCs were transplanted into triploid white catfish between 4.0 to 6.0 DPH (27.4 ± 0.4°C). Overall, these findings allow enhanced efficiency of commercializing xenogenic catfish carrying gametes of either blue catfish or channel catfish.

Practical diets for California yellowtail, Seriola dorsalis: Use of advanced soybean meal products on growth performance, body composition, intestinal morphology, and immune gene expression.

California yellowtail (CYT), Seriola dorsalis, is a promising candidate for aquaculture due to its rapid growth and high-quality flesh, particularly in markets like Japan, Australia, China, and the United States. Soy protein has shown success as a replacement for marine protein sources in CYT diets, reducing fishmeal levels, though concerns about potential intestinal inflammation persist with the inclusion of solvent-extracted soybean meal. To address this, processing strategies like fractionation, enzymatic treatment, heat treatment, and microbial fermentation have been employed to mitigate the negative impacts of soybean meal on fish nutrition and immune systems. This study focuses on optimizing soybean meal inclusion levels by incorporating advanced soy variants into CYT diets. The eight-week feeding trial, conducted in a recirculation system, featured six diets with sequential inclusion levels (0, 50, 100%) of high protein low oligosaccharide soybean meal (Bright Day, Benson Hill, St Louis, MO) and enzyme-treated soybean meal (HP 300, Hamlet Protein Inc., Findlay, OH), replacing solvent-extracted soybean. The study compares these formulations against a soy-free animal protein-based diet. At the end of the trial, fish were sampled for growth performance, body proximate composition, intestinal morphology, and immune response from gut samples. Results showed consistent FCR (P = 0.775), weight gain (P = 0.242), and high survival rate (99.4 ± 0.5%) among dietary treatments (P>0.05). Histological evaluations revealed no gut inflammation and gene expression analysis demonstrated no significant variations in immune, physiological, and digestive markers apn (P = 0.687), mga (P = 0.397), gpx1 (P = 0.279), atpase (P = 0.590), il1ß (P = 0.659). The study concludes that incorporating advanced soybean meal products, replacing up to 20% of fishmeal does not negatively affect CYT's growth and intestinal health. This suggests that all three soy sources, contributing 35% of total protein (15.4 g 100 g-1 diet), can be included in practical diets without compromising CYT's intestinal integrity or growth. These findings have positive implications for the commercial production of CYT and future research on the incorporation of plant-based proteins in aquaculture diets.

Investigating the optimum stocking density of tilapia (Oreochromis niloticus) for intensive production focused to in-pond raceway system.

The primary objective of this trial was to study the effects of stress caused by stocking density in tilapia (Oreochromis niloticus) cultured in the in-pond raceway system (IPRS). Fingerlings (Initial body weight = 30.00 ± 1.20 g) were reared at different stocking densities i.e. low stocking density (n = 13,000; 1.77 kg/m3), medium stocking density (MSD) (n = 17,000; 2.32 kg/m3), and high stocking density (HSD) (n = 21,000; 2.86 kg/m3), all confined within the raceways of the IPRS. Each group was studied in triplicates. The observed growth revealed a regression in the HSD treatment, evident in its reduced weight gain per fish per day, in contrast to other treatments. Survival rate across all treatments was above 99%. Notably, the HSD treatment exhibited an elevated level of cortisol; however, this intensified crowding stress did not significantly undermine the nutritional value of the fish in HSD and other experimental treatments. Furthermore, an elevation in the levels of superoxide dismutase, catalase, and glutathione peroxidase was noted within the HSD treatment in comparison to the other treatments to reduce the damage caused by reactive oxygen species. As the trial proceeded, functions of digestive enzymes like amylase, protease, and lipase diminished consistently across all treatments. This could possibly signify a deliberate redirection of energy resources toward stress alleviation rather than the usual digestive processes. In summation, it can be reasonably deduced that a stocking density of approximately 2.32 kg/m3 (MSD) emerges as the optimal threshold for effectively cultivating tilapia within an intensive aquaculture framework.

The growth performance and parasite load of angelfish juveniles Pterophyllum scalare kept at different stocking densities in two rearing systems.

This study evaluated the growth performance and parasite load of angelfish juveniles Pterophyllum scalare kept at different stocking densities using two rearing systems. The experiment was conducted in a factorial design (4x2) with four stocking densities (0.1, 0.4, 0.7, and 1.0 g/L), two type of aquarium tanks (glass and ceramic aquariums), and four replicates. The experiment lasted 60 days using 148 juvenile fish (3.05 ± 0.09 g) randomly placed in 32 aquariums (50 L) equipped with filters and aeration. All fish received two meals a day ad libitum (8:00 and 16:00). Water quality parameters such as temperature, dissolved oxygen, pH, and total ammonia were measured. At the end of the experiment, all fish were measured and weighed to determine growth performance and then subjected to parasitological analysis. The data were analyzed with a two-way ANOVA with post-hoc Tukey test (p<0.05). No effects on growth performance at different stocking densities were observed. However, there was an increase in Capillaria pterophylli infestation in the high stocking density within ceramic aquariums. Thus, this study recommends the use of 1.0 g/L for the intensive aquaculture system of freshwater angelfish, and applying cleaning management to avoid parasite infestation, particularly in ceramic aquariums.

Growth and survival of Betta splendens fed microbial aggregates from ex-situ biofloc technology (BFT).

The betta (Betta splendens) is a carnivorous ornamental species that requires live food. In this study, we evaluated the growth and survival of Betta splendens fingerlings fed ex-situ biofloc (BFT). A total of 375 fingerlings (8.3 ± 4.1 mg and 0.8 ± 0.14 cm) were distributed into 15-L plastic tanks (n=25), with five replicates for each of the following treatments: (i) exclusive live food (LF100), (ii) exclusive biofloc (BFT100), (iii) live food supplemented with biofloc (LF100-BFT100), and a reduction of 15% (iv) and 30% (v) in live food with biofloc supplementation (LF85-BFT100 and LF70-BFT100, respectively). The fish were fed newly hatched brine shrimp (Artemia spp. nauplii) and/or fresh biofloc, twice daily, over a 16-day experimental period. Dietary supplementation of Beta splendens with ex-situ BFT (LF100-BFT100, LF85-BFT100, and LF70-BFT100) resulted in a similar final weight (P>0.05) compared to exclusive brine shrimp (LF100). Regarding total length, the fish in the LF100-BFT100 group were superior to those in the LF100 group (P<0.05). The LF100 and LF100-BFT100 groups showed survival rates of 100% and 98.4%, respectively, which were not statistically different (P>0.05). For all analyzed variables, fish exclusively fed biofloc (BFT100) exhibited unfavorable performance. These results indicate that dietary ex-situ biofloc supplementation, without reducing the supply of live food, can be an interesting alternative in the rearing of Betta splendens since biofloc improves growth performance and maintains a good survival rate.

Automatic Shrimp Fry Counting Method Using Multi-Scale Attention Fusion.

Shrimp fry counting is an important task for biomass estimation in aquaculture. Accurate counting of the number of shrimp fry in tanks can not only assess the production of mature shrimp but also assess the density of shrimp fry in the tanks, which is very helpful for the subsequent growth status, transportation management, and yield assessment. However, traditional manual counting methods are often inefficient and prone to counting errors; a more efficient and accurate method for shrimp fry counting is urgently needed. In this paper, we first collected and labeled the images of shrimp fry in breeding tanks according to the constructed experimental environment and generated corresponding density maps using the Gaussian kernel function. Then, we proposed a multi-scale attention fusion-based shrimp fry counting network called the SFCNet. Experiments showed that our proposed SFCNet model reached the optimal performance in terms of shrimp fry counting compared to CNN-based baseline counting models, with MAEs and RMSEs of 3.96 and 4.682, respectively. This approach was able to effectively calculate the number of shrimp fry and provided a better solution for accurately calculating the number of shrimp fry.

The development of multiplex PCR assays for the rapid identification of multiple Saccostrea species, and their practical applications in restoration and aquaculture.

BACKGROUND: The ecology and biology of oysters (Ostreidae) across the tropics is poorly understood. Morphological plasticity and shared characteristics among oysters have resulted in the misidentification of species, creating challenges for understanding basic species-specific biological information that is required for restoration and aquaculture. Genetic barcoding has proven essential for accurate species identification and understanding species geographic ranges. To reduce the costs of molecular species identification we developed multiplex assays using the cytochrome c oxidase subunit I (COI or cox1) barcoding gene for the rapid identification of five species of oysters within the genus Saccostrea that are commonly found in Queensland, Australia: Saccostrea glomerata, Saccostrea lineage B, Saccostrea lineage F, Saccostrea lineage G, and Saccostrea spathulata (lineage J). RESULTS: Multiplex assays were successful in species-specific amplification of targeted species. The practical application of these primers was tested on wild spat collected from a pilot restoration project in Moreton Bay, Queensland, with identified species (S. glomerata, lineage B and lineage G) validated by Sanger sequencing. DNA sampling by extraction of oyster pallial fluid was also tested on adult oysters collected from the Noosa estuary in Queensland to assess whether oysters were able to be identified non-destructively. DNA concentrations as low as 1 ng/ µL still amplified in most cases, allowing for identification, and mortality at 6 weeks post pallial fluid collection was low (3 out of 104 sampled oysters). CONCLUSION: These multiplex assays will be essential tools for species identification in future studies, and we successfully demonstrate their practical application in both restoration and aquaculture contexts in Queensland. The multiplex assays developed in this study outline easily replicable methods for the development of additional species-specific primer sets for the rapid identification of other species of Saccostrea found across the Indo-Pacific, which will be instrumental in unravelling the taxonomic ambiguities within this genus in tropical regions.

Influence of taurine on the zootechnical performance and health parameters of juvenile Nile tilapia in a recirculating aquaculture system.

Taurine is considered a conditionally essential amino acid for fish, so its supplementation may improve feed conversion. This study evaluated the supplementation of taurine on growth performance, hematological and immunological parameters, production costs, and survival of Nile tilapia (Oreochromis niloticus) juveniles raised in a recirculating aquaculture system (RAS). A control diet was formulated with 360 g kg-1 of crude protein without fish meal and without taurine supplementation (Control). From the control diet, another diet supplemented with 9.7 g of taurine per kg of feed (Taurine) was produced. Fish fed diet supplemented with taurine had lower daily average weight gain and final average weight compared to the control diet (p < 0.05). It was observed that taurine had no influence on condition factor, survival, or hemato-immunological parameters of Nile tilapia juveniles, but there was a higher mean corpuscular volume and greater nitrogen retention in fish from the control group (p < 0.05). It is concluded that Nile tilapia juveniles do not benefit from taurine supplementation in RAS, even when fed diet containing plant-based protein sources.

Influence of stocking density on the growth, immune and physiological responses, and cultivation environment of white-leg shrimp (Litopenaeus vannamei) in biofloc systems.

Biofloc (BF) stands out as a promising system for sustainable shrimp farming. Optimizing various culture conditions, such as stocking density, carbohydrate source, and feeding management, is crucial for the widespread adoption of the BF system. This study compares the growth performance of white-leg shrimp (Litopenaeus vannamei) in culture ponds at low density (LD) with 50 organisms/m2 and high density (HD) with 200 organisms/m2. Post-larvae of white-leg shrimp were stocked for 16 weeks in both LD and HD groups. The LD group exhibited a superior survival rate, growth rate, and feed consumption compared to the HD group. The BF from the LD system recorded a significantly higher protein content (16.63 ± 0.21%) than the HD group (15.21 ± 0.34%). Heterotrophic bacterial counts in water did not significantly differ with stocking density. However, Vibrio count in water samples was higher in the HD group (3.59 ± 0.35 log CFU/mL) compared to the LD group (2.45 ± 0.43 log CFU/mL). The whole shrimp body analysis revealed significantly higher protein and lipid content in the LD group. In contrast, the total aerobic bacterial count in shrimp from the HD group was high, with the identification of Salmonella enterica ssp. arizonae. Additionally, Vibrio counts in shrimp samples were significantly higher in the HD group (4.63 ± 0.32 log CFU/g) compared to the LD group (3.57 ± 0.22 log CFU/g). The expression levels of immune-associated genes, including prophenoloxidase, transglutaminase, penaiedin 3, superoxide dismutase, lysozyme, serine proteinase, and the growth-related gene ras-related protein (rap-2a), were significantly enhanced in the LD group. Conversely, stress-related gene expression increased significantly in the HD group. Hepatopancreases amylase, lipase, and protease were higher in the LD group, while trypsin activity did not differ significantly. Antioxidant enzyme activity (catalase, glutathione, glutathione peroxidase, and superoxide dismutase) significantly increased in the LD group. The histological structure of hepatopancreas, musculature, and female gonads remained similar in both densities. However, negative effects were observed in the gills' histology of the HD group. These results suggest that increasing stocking density is associated with significantly negative biological, microbial, and physiological effects on white-leg shrimp under the BF system.

Vegetable omega-3 and omega-6 fatty acids differentially modulate the antiviral and antibacterial immune responses of Atlantic salmon.

The immunomodulatory effects of omega-3 and omega-6 fatty acids are a crucial subject of investigation for sustainable fish aquaculture, as fish oil is increasingly replaced by terrestrial vegetable oils in aquafeeds. Unlike previous research focusing on fish oil replacement with vegetable alternatives, our study explored how the omega-6 to omega-3 polyunsaturated fatty acid (PUFA) ratio in low-fish oil aquafeeds influences Atlantic salmon's antiviral and antibacterial immune responses. Atlantic salmon were fed aquafeeds rich in soy oil (high in omega-6) or linseed oil (high in omega-3) for 12 weeks and then challenged with bacterial (formalin-killed Aeromonas salmonicida) or viral-like (polyriboinosinic polyribocytidylic acid) antigens. The head kidneys of salmon fed high dietary omega-3 levels exhibited a more anti-inflammatory fatty acid profile and a restrained induction of pro-inflammatory and neutrophil-related genes during the immune challenges. The high-omega-3 diet also promoted a higher expression of genes associated with the interferon-mediated signaling pathway, potentially enhancing antiviral immunity. This research highlights the capacity of vegetable oils with different omega-6 to omega-3 PUFA ratios to modulate specific components of fish immune responses, offering insights for future research on the intricate lipid nutrition-immunity interplay and the development of novel sustainable low-fish oil clinical aquaculture feeds.

Effects of different stocking densities on growth, nutritional quality, stress and antioxidant response in Labeo rohita; cultured in in-pond raceway system.

A 171-day long experimental trial was undertaken to study intricate physiological response of rohu (Labeo rohita) under stress caused by high stocking density in In-pond raceways system (IPRS). Fingerlings of rohu (initial body weight: 250 ± 1.20 g) were cultured at three different stocking densities; low density (LD) (2.27 kg/m3), medium density (MD) (3.79 kg/m3) and high density (HD) (5.30 kg/m3) in raceways of IPRS production system. Each treatment was in triplicate. Fish growth exhibited a decline in HD treatment statistically as its average weight gain/fish/day was 4.21 g as compared to MD (4.82 g) and LD (4.74 g). Nutritional profile of rohu indicated by the content of crude protein, fatty acids, and profile of amino acids was up to the set dietary benchmarks. Survival rate of fish in all the treatment groups was greater than 99%. The elevated cortisol levels observed in the HD treatment in contrast to the other treatments suggested the presence of stress. The levels of superoxide dismutase, catalase and glutathione peroxidase were also higher in HD as compared to other treatments. However, there were no difference in the level of MDA between the three treatments. Activity of amylase, protease was significantly different in treatment whereas the difference in lipase activity was found to be insignificant. It can be concluded that medium stocking density i.e. 3.79 kg/m3 outperformed the high density (5.30 kg/m3) in different aspects of this study. Nevertheless, additional research is imperative to ascertain whether any intermediate stocking density between medium (3.79 kg/m3) and high (5.30 kg/m3) such as 4 kg/m3, 4.5 kg/m3, or 5 kg/m3, could potentially serve as suitable options for rohu. It is also suggested that brood stock of rohu should be genetically improved to obtain stress resilient fingerlings which will perform better at high stocking density at large scale production level.

Evaluation of Alternative Sources of Proteins and Other Nutrients with Potential Applications in Fish Nutrition.

The European Union's (EU) agricultural self-sufficiency is challenged by its reliance on imported plant proteins, particularly soy from the Americas, contributing to deforestation and greenhouse gas emissions. Addressing the EU's protein deficit, this study evaluates alternative protein sources for aquaculture, focusing on their nutritional value, elemental content, and polycyclic aromatic hydrocarbons (PAHs). Protein flours from gastropods (Helix pomatia, Arion lusitanicus, Arion vulgaris) and their hepatopancreas, along with plant-based proteins from food industry by-products (oilcakes, coffee grounds, spent brewer's yeast), were analyzed. Results revealed that snail flour contained the highest protein content at 59.09%, significantly outperforming hepatopancreas flour at 42.26%. Plant-based proteins demonstrated substantial nutritional value, with coffee grounds flour exhibiting a remarkable protein content of 71.8% and spent brewer's yeast flour at 57.9%. Elemental analysis indicated high levels of essential minerals such as magnesium in hepatopancreas flour (5719.10 mg/kg) and calcium in slug flour (48,640.11 mg/kg). However, cadmium levels in hepatopancreas flour (11.45 mg/kg) necessitate caution due to potential health risks. PAH concentrations were low across all samples, with the highest total PAH content observed in hepatopancreas flour at 0.0353 µg/kg, suggesting minimal risk of PAH-related toxicity. The analysis of plant-based protein sources, particularly oilcakes derived from sunflower, hemp, flax, and pumpkin seeds, revealed that these by-products not only exhibit high protein contents but present a promising avenue for enhancing the nutritional quality of feed. This study underscores the potential of utilizing gastropod and plant-based by-products as sustainable and nutritionally adequate alternatives to conventional feeds in aquaculture, contributing to the EU's environmental sustainability goals.

A novel multitrophic biofloc technology for duckweed and Megalobrama amblycephala integrated culture: Improving nutrient utilization and animal welfare.

Biofloc technology (BFT) is an eco-friendly aquaculture model that utilizes zero-exchange water. In this study, we investigated the integration of duckweed into BFT in an effort to enhance nitrogen, phosphorus, and carbon utilization and to improve animal welfare for cultivating Megalobrama amblycephala. The experiment spanned 75 days, comparing a group of M. amblycephala supplemented with duckweed (DM) to a control group (CG) with no supplementation, where duckweed consumption relied solely on the feeding behavior of the fish. The concentrations of nitrate, total nitrogen, and phosphorus accumulation were lower in the DM than in the CG from day 45 onwards, with differences of 16.19, 26.90, and 1.45 mg/L, respectively, at the end of the experiment. The DM showed simultaneous increases of 5.77, 11.20, and 5.07 % in the absolute utilization of nitrogen, phosphorus, and carbon, respectively. The abundance of TM7a (10.27 %), linked to nitrate absorption, became the dominant genus in the water of the DM. Additionally, the abundance of Cetobacterium, associated with carbohydrate digestion, was significantly higher in gut of the DM (23.83 %) than in the gut of CG (1.24 %, P < 0.05). Supplementing the diet of M. amblycephala with duckweed improved digestion and antioxidant enzyme activity. Transcriptome data showed that duckweed supplementation resulted in an increase in the expression of genes related to protein digestion and absorption and carbohydrate metabolism in M. amblycephala, and analysis of the significantly enriched pathways further supported improved antioxidant capacity. Based on the above results, we concluded that as M. amblycephala consumes more duckweed, the differences in nitrogen and phosphorus levels between the DM and CG would continue to increase, along with a simultaneous increase in fixed carbon. Thus, this study achieved the goal of recycling BFT resources and improving animal welfare by integrating duckweed.

Chlorella vulgaris algae ameliorates chlorpyrifos toxicity in Nile tilapia with special reference to antioxidant enzymes and Streptococcus agalactiae infection.

BACKGROUND: Chlorpyrifos (CPF) is a widely used pesticide in the production of plant crops. Despite rapid CPF biodegradation, fish were exposed to wastewater containing detectable residues. Recently, medicinal plants and algae were intensively used in aquaculture to replace antibiotics and ameliorate stress impacts. METHODS AND RESULTS: An indoor experiment was conducted to evaluate the deleterious impacts of CPF pollution on Nile tilapia health and the potential mitigation role of Chlorella vulgaris algae. Firstly, the median lethal concentration LC50 - 72 h of CPF was determined to be 85.8 µg /L in Nile tilapia (35.6 ± 0.5 g body weight) at a water temperature of 27.5 °C. Secondly, fish were exposed to 10% of LC50 - 72 h for six weeks, and tissue samples were collected and examined every two weeks. Also, Nile tilapia were experimentally infected with Streptococcus agalactiae. Exposed fish were immunosuppressed expressed with a decrease in gene expressions of interleukin (IL) 1ß, IL-10, and tumor necrosis factor (TNF)-α. Also, a decline was recorded in glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) gene expression in the head kidney tissue. A high mortality rate (MR) of 100% was recorded in fish exposed to CPF for six weeks and challenged with S. agalactiae. Fish that received dietary C. vulgaris could restore gene expression cytokines and antioxidants compared to the control. After six weeks of CPF exposure, fish suffered from anemia as red blood cell count (RBCs), hemoglobin (Hb), and packed cell volume (PCV) significantly declined along with downregulation of serum total protein (TP), globulin (GLO), and albumin (ALB). Liver enzymes were significantly upregulated in fish exposed to CPF pollution, alanine aminotransferase (ALT) (42.5, 53.3, and 61.7 IU/L) and aspartate aminotransferase (AST) (30.1, 31.2, and 22.8) after 2, 4, and 6 weeks, respectively. On S. agalactiae challenge, high MR was recorded in Nile tilapia exposed to CPF (G3) 60%, 60%, and 100% in week 2, week 4, and week 6, and C. vulgaris provided a relative protection level (RPL) of 0, 14.29, and 20%, respectively. CONCLUSIONS: It was concluded that CPF pollution induces immunosuppressed status, oxidative stress, and anemic signs in Nile tilapia. In contrast, C. vulgaris at a 50 g/kg fish feed dose could partially ameliorate such withdrawals, restoring normal physiological parameters.

Sampling time for different matrices in stress assessment of farmed Atlantic salmon post-smolt.

The sustainability of commercial aquaculture production depends critically on prioritizing fish welfare management. Besides monitoring welfare parameters such as fish behaviour and water quality, fish stress level can also provide a reliable measure of the welfare status of farmed fish. Cortisol and 5 of its metabolites (5ß-THF, cortisone, 5ß-DHE, 5ß-THE, ß-cortolone) were previously identified by the authors as suitable stress biomarkers of farmed Atlantic salmon. Based on this knowledge, the present study aimed to investigate the time-related dynamics of these metabolites in plasma, skin mucus, bile and faeces over a 72 h- period. The objective was to determine the optimal sampling time for each matrix and to understand the clearance pathway of these metabolites following stress. An experiment was carried out using a total of 90 Atlantic salmon with an average weight of 438 (±132) g. The average sea temperature was 6.9 °C during the experimental period. A control group of 10 fish was first collected before the remaining 80 fish were submitted to a stress of netting and subsequent relocation into two separate cages. From each of these two stress groups, 10 fish were sampled at 1 h, 2 h, 4 h, 6 h and 12 h, 24 h, 48 h, 72 h after the stress event respectively. The concentrations of cortisol and its metabolites were measured at each of the sampling timepoint. The results demonstrated that plasma cortisol metabolites reached the highest concentration 4 h after stress and remained elevated despite the slight decrease for the remaining timepoints. The peak level was observed at 12 h post-stress in skin mucus and 24 h in bile and faeces. The findings suggest that these timepoints are the optimal for sampling Atlantic salmon post-smolt following stressful events in acute stress studies. Furthermore, the results reveal that analysing cortisol and its metabolites, both in free and conjugated forms, rather than free cortisol provides greater flexibility as their concentrations are less affected by sampling procedure. This study confirms the appropriateness of skin mucus and faeces as less-invasive sample matrices for fish stress evaluation and provides a basis for further developing low invasive tools for monitoring the welfare of farmed salmonid.

Analysis of brominide disinfection by-products (DBPs) in aquaculture water using ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-tof/MS).

BACKGROUND: halogenic disinfectants have been shown to produce toxic and carcinogenic disinfection by-products in the water disinfection process. Dibromohydantoin (DBDMH) is a commonly used water disinfectant in aquaculture. Aquaculture water has more complex matrix, and the analytical method for disinfection by-products (DBPs) have not been reported. Since the content of DBPs is related to the external conditions such as ultraviolet irradiation, temperatures, pH and humic acid. The semi-target screening method for mainly DBPs based on tracing mass spectrometry fragments of bromide and accurate mass of high resolution mass spectrometry was established by ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-tof/MS). Br-DBPs as a important class of DBPs from DBDMH, which quantification analysis methods were developed based on accurate mass of high resolution mass spectrometry. METHODS: through screening method to identify unknown Br-DBPs and quantitative analysis of the typical 4-bromophenol by-product of accurate mass was established. The conditions of the instrument parameters of mass spectrometry and SPE sample preparation procedure in complex real sample were optimized. The high efficiency method was demonstrated for the determination of Br-DBPs with a good linear correlation (R2 = 0.999) in the range of 0.500-200 µg L-1 and limit of detections (LODs) and limit of quantifications (LOQs) were 0.0250 ng L-1 and 0.0834 ng L-1, respectively. CONCLUSION: the developed screening and quantification analytical strategy for Br-DBPs is rapid, accurate and sensitivity applicable for environmental in aquaculture water monitoring.

Real-world waste dispersion modelling for benthic integrated multi-trophic aquaculture.

In real-world situations, marine fish farms accommodate multiple fish species and cohorts within the farm, leading to diverse farm layouts influenced by cage dimensions, configurations, and intricate arrangements. These cage management practices are essential to meet production demands, however, farm-level complexities can impact model predictions of waste deposition and benthic impact near fish cages. This is of particular importance when the cages are used for integrated multi-trophic aquaculture (IMTA) with benthic feeders, where this waste not only affects environmental conditions but also provides a potential food source. The Cage Aquaculture Particulate Output and Transport (CAPOT) model incorporated multiple species, cohorts, and cage arrangements to estimate waste distribution from a commercial fish farm in the Mediterranean between October 2018 and July 2019. This spreadsheet model estimated dispersion for individual fish cages using a grid resolution of 5 m x 5 m. The study categorized discrete production periods for each fish cage every month, aligning with intermittent changes in biomass and food inputs due to different cage management practices throughout production. This approach facilitated the use of detailed input data and enhanced model representativeness by considering variations in cage biomass, food types, settling velocities, and configurations. Model outputs, represented in contour plots, indicated higher deposition directly below fish cages that varied monthly throughout fish production cycles. Deposition footprints reflected changes in cage biomass, food inputs, and farm-level practices reflecting this real-world scenario where aquaculture does not follow a production continuum. Moreover, cohort dynamics and cage movements associated with the cage management practices of the fish farm influenced the quantity and fate of wastes distributed around fish cages, revealing variability in deposition footprints. Clearly, these findings have important implications for the design of benthic IMTA systems, with species such as sea cucumber and polychaetes. Variability in waste deposition creates challenges in identifying where the benthic organisms should be placed to allow optimal uptake of waste to meet their food requirements and increase survivability. Evidently, models have an important role to play and this study emphasizes the need for representative input data to describe actual food inputs, cage biomass changes, and management practices for more representative farm-scale modelling and essentially to improve particulate waste management. To effectively mitigate benthic impacts through IMTA, models must quantify and resolve particulate waste distribution and impact around fish farms to maintain a balanced system with net removal of wastes. Resolving farm-level complexities provides vital information about the variability of food availability and quality for extractive organisms that helps improve recycling of organic wastes in integrated systems, demanding a more representative modelling approach.

Could fish aggregation at ocean aquaculture augment wild populations and local fisheries?

The global population consumes more seafood from aquaculture today than from capture fisheries and although the aquaculture industry continues to grow, both seafood sectors will continue to be important to the global food supply into the future. As farming continues to expand into ocean systems, understanding how wild populations and fisheries will interact with farms will be increasingly important to informing sustainable ocean planning and management. Using a spatially explicit population and fishing model we simulate several impacts from ocean aquaculture (i.e., aggregation, protection from fishing, and impacts on fitness) to evaluate the mechanisms underlying interactions between aquaculture, wild populations and fisheries. We find that aggregation of species to farms can increase the benefits of protection from fishing that a farm provides and can have greater impacts on more mobile species. Splitting total farm area into smaller farms can benefit fishery catches, whereas larger farms can provide greater ecological benefits through conservation of wild populations. Our results provide clear lessons on how to design and co-manage expanding ocean aquaculture along with wild capture ecosystem management to benefit fisheries or conservation objectives.