Enhancing Florida red tilapia aquaculture: biofloc optimization improves water quality, pathogen bacterial control, fish health, immune response, and organ histopathology across varied groundwater salinities.

Freshwater scarcity poses challenges to aquaculture worldwide, including countries like Egypt. In this study, we investigate the feasibility of integrating underground saline water (USW) with varying salinities into a Biofloc (BFT) system for desert mariculture of Florida red tilapia (FRT) and its impacts on water quality, fish performance and health. Four BFT treatments (C/N ratio = 15) were examined in triplicate using four salinity levels 0 ppt, 12 ppt, 24 ppt and 36 ppt, expressed as S0, S12, S24 and S36, respectively. For 75 days, a total of 12 fiberglass tanks (each 250 L-1 water) were used to store FRT fry (average weight of 1.73 ± 0.01 g/fish). The fish were fed an experimental diet (protein/fat = 30/5) and an additional carbon source of rice bran. The results revealed that group S12 showed better growth indicators, higher survival rate, lower FCR, and lower ammonia levels, while group S0 exhibited lower growth indicators (final weight, weight gain, and specific growth rate) than all groups. The serum kidney, liver, and antioxidant indices performed better in the S12 group. At 12 ppt, the immune-related parameter (IgM) increased by 22.5%, while the stress parameter (cortisol) decreased by 40.8% compared to the S0 group. The liver and intestinal histopathological results revealed that the S12 and S24 groups performed better. Pathogenic bacterial load counts favored the S24 group, which had the lowest number among the groups studied. The recommended salinity for FRT cultivation in USW and BFT is 19.94-20 ppt, determined by polynomial regression of FW and FCR.

Comparative impact of replacing fish meal with azolla on growth, water quality, and physiology of red tilapia fingerlings at varying salinities.

A 210-day experiment to assess the efficacy of substituting azolla plant powder at levels of 0, 20, 40, and 60% for fish meal on red tilapia fingerlings (RTF, initial weight of 18.23 ± 0.12 g) performance under salinity levels of 5, 18, and 28ppt. Among the various conditions, RTF-fed 20% azolla at 28 and 5ppt salinity showcased the highest specific growth rate (SGR), whereas the lowest SGR was observed in fish-fed 60% azolla at 5ppt salinity. Upon azolla incorporation, noteworthy elevations in phytoplankton, zooplankton, dissolved oxygen (DO), pH, NH3, and NO3 were noted and conversely, azolla introduction led to decreased NH4 and NO2 concentrations in all salinity levels. Further, a significant (p < 0.05) interaction between azolla levels and water salinity (S×A) significantly impacted the hematological parameters of RTF. The highest levels of superoxide dismutase (SOD), catalase (CAT), and total protein (TP) were found in RTF-fed 20% azolla at 28ppt salinity, while the lowest CAT and TP levels occurred in RTF-fed 60% azolla at 5ppt salinity. The highest aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were recorded in the RTF group fed 60% azolla at 5ppt salinity, with the lowest values seen in the group given 20% azolla at 28ppt salinity. RTF fed a 20% azolla diet at 18ppt salinity exhibited the highest lysozyme value, in contrast to the lowest value observed in the RTF group fed the control diet at 18ppt salinity. In conclusion, this study recommends the utilization of azolla at inclusion levels ranging from 20 to 40%, as it has the potential to notably enhance the immune system and elevate the survival rate of RTF.

Plant Growth-Promoting Rhizobacteria as Tools to Improve the Growth of Kohlrabi (Brassica oleracea var. gongylodes) Plants in an Aquaponics System.

The use of nitrogen as a fertilizer can be highly risky when used excessively, and it is therefore necessary to find novel techniques to reduce its use. Aquaponics reduces the use of synthetic fertilizers and water, and the leaching of nitrate into the environment. One way to avoid problems due to a reduction in nitrogen availability could be the use of plant growth promoting rhizobacteria (PGPR). This study examines the effect of PGPR on kohlrabi plants grown with a traditional nutrient solution (100S), in combination with "fish water" (50F/50D), or with a supplement of synthetic fertilizers (50F/50D + S). Two formulations were used: T1 (Azospirillum brasilense and Pantoea dispersa) and T2 (Azotobacter salinestris). Irrigation with 50F/50D caused a reduction in several of the measured parameters. The combined application of 50F/50D with T1 attenuated the negative effects. T2 did not present significant effects on the parameters measured. The results obtained with 50F/50D + S hardly showed differences with the 100S. Thus, by irrigating with 50F/50D + S, we were able to maintain the yields while reducing fertilizer use and water. The combined use of T1 and 50F/50D was also positive; however, it would be necessary to continue adjusting the amount of nitrate supplied to maintain production.

Systemic and mucosal immune responses in red tilapia (Oreochromis sp.) following immersion vaccination with a chitosan polymer-based nanovaccine against Aeromonas veronii.

A mucoadhesive chitosan polymer-based nanoplatform has been increasingly recognized as an effective mucosal vaccine delivery system for fish. The present study aimed to investigate the effectiveness of immersion vaccination with a chitosan polymer-based nanovaccine to elicit an immune response in serum and mucus of red tilapia and evaluate its protective efficacy after immersion challenge with a heterogenous strain of Aeromonas veronii UDRT09. Six hundred red tilapia (22 ± 1.8 g) were randomly allocated into four experimental groups: control, empty-polymeric nanoparticle (PC), formalin-killed vaccine (FKV), and chitosan polymer-based nanovaccine (CS-NV) in triplicate. The specific IgM antibody levels and their bactericidal activity were assessed in serum and mucus for 28 days after immersion vaccination and followed by immersion challenge with A. veronii. The immersion vaccine was found to be safe for red tilapia, with no mortalities occurring during the vaccination procedure. The specific IgM antibody levels and bactericidal activity against A. veronii in both serum and mucus were significantly higher in red tilapia vaccinated with CS-NV compared to the FKV and control groups at all time points. Furthermore, the serum lysozyme activity, ACH50, and total Ig levels demonstrated a significant elevation in the groups vaccinated with CS-NV compared to the FKV and control groups. Importantly, the Relative Percentage Survival (RPS) value of the CS-NV group (71 %) was significantly higher than that of the FKV (15.12 %) and PC (2.33 %) groups, respectively. This indicates that the chitosan polymer-based nanovaccine platform is an effective delivery system for the immersion vaccination of tilapia.

l-Threonine production from whey and fish hydrolysate by E. coli ATCC® 21277TM.

In this work production of l-threonine by Escherichia coli ATCC® 21277™ has been studied using a mixture of alternative low-cost substrates, which are recognized to be a major pollution problem. Whey was used as the primary carbon source, whereas Red Tilapia (Oreochromis sp.) viscera hydrolysates constituted the nitrogen source. A Box-Behnken Design was used for optimizing l-threonine and biomass production, using temperature and glucose, whey, and Red Tilapia (Oreochromis sp.) viscera hydrolysate contents as factors. Results indicate that biomass production is affected by the concentration of hydrolysate and temperature. On the other hand, l-threonine production is affected by concentration of whey, hydrolysate, and temperature. In this context, it was possible to maximize l-threonine production, but with a detriment on biomass production. The optimal conditions for biomass and l-threonine maximization (after 24 h) were identified and validated experimentally, resulting in biomass and l-threonine production of 0.767 g/L and 0.406 g/L, respectively. This work has shown the technical feasibility of using whey and Red Tilapia (Oreochromis sp.) viscera hydrolysates for the production of l-threonine by E. coli ATCC® 21277TM. Finally, the complications associated to the use of these low-cost complex substrates for the production of l-threonine by E. coli, suggest that more in detail studies (i.e. at the metabolic level) are required in order to propose strategies to increase the process productivity, before its scale up. This is a first step in our long-term goal of developing a production process for i) dealing with the pollution problems caused by those wastes, and ii) strengthen the milk and fish industries which are important poles of the Colombian economy.

Basil functional and growth responses when cultivated via different aquaponic and hydroponics systems.

Background: Aquaponics is an innovative farming system that combines hydroponics and aquaculture, resulting in the production of both crops and fish. Decoupled aquaponics is a new approach introduced in aquaponics research for the elimination of certain system bottlenecks, specifically targeting the optimization of crops and fish production conditions. The aquaponics-related literature predominantly examines the system's effects on crop productivity, largely overlooking the plant functional responses which underlie growth and yield performance. The aim of the study was the integrated evaluation of basil performance cultivated under coupled and decoupled aquaponic systems compared with a hydroponic one, in terms of growth and functional parameters in a pilot-scale aquaponics greenhouse. Methods: We focused on the efficiency of the photosynthetic process and the state of the photosynthetic machinery, assessed by instantaneous gas exchange measurements as well as photosynthetic light response curves, and in vivo chlorophyll a fluorescence. Light use efficiency was estimated through leaf reflectance determination. Photosynthetic pigments content and leaf nutritional state assessments completed the picture of basil functional responses to the three different treatments/systems. The plant's functional parameters were assessed at 15-day intervals. The experiment lasted for two months and included an intermediate and a final harvest during which several basil growth parameters were determined. Results: Coupled aquaponics resulted in reduced growth, which was mainly ascribed to sub-sufficient leaf nutrient levels, a fact that triggered a series of negative feedbacks on all aspects of their photosynthetic performance. These plants experienced a down-regulation of PSII activity as reflected in the significant decreases of quantum yield and efficiency of electron transport, along with decreased photosynthetic pigments content. On the contrary, decoupled aquaponics favored both growth and photochemistry leading to higher light use efficiency compared with coupled system and hydroponics, yet without significant differences from the latter. Photosynthetic light curves indicated constantly higher photosynthetic capacity of the decoupled aquaponics-treated basil, while also enhanced pigment concentrations were evident. Basil functional responses to the three tested production systems provided insights on the underlying mechanisms of plant performance highlighting key-points for systems optimization. We propose decoupled aquaponics as an effective system that may replace hydroponics supporting high crops productivity. We suggest that future works should focus on the mechanisms involved in crop and fish species function, the elucidation of which would greatly contribute to the optimization of the aquaponics productivity.

Masculinization of Red Tilapia (Oreochromis spp.) Using 17α-Methyltestosterone-Loaded Alkyl Polyglucosides Integrated into Nanostructured Lipid Carriers.

The aim of the present study was to optimize a masculinization platform for the production of all-male red tilapia fry by oral administration of 30 and 60 ppm of MT and alkyl polyglucoside nanostructured lipid carriers (APG-NLC) loaded with MT, respectively, for 14 and 21 days. The characterization, encapsulation efficiency and release kinetics of MT in lipid-based nanoparticles were assessed in vitro. The results showed that the MT-loaded nanoparticles were spherical, ranging from 80 to 125 nm in size, and had a negative charge with a narrow particle distribution. The APG-NLC loaded with MT provided higher physical stability and encapsulation efficacy than the NLC. The release rate constants of MT from MT-NLC and MT-APG-NLC were higher than those of free MT, which is insoluble in aqueous media. There was no significant difference in survival between the fish administered MT or the those fed orally with MT-APG-NLC fish. According to the logistic regression analysis, the sex reversal efficacy of MT-APG-NLC (30 ppm) and MT (60 ppm), resulted in significantly higher numbers of males after 21 days of treatment compared with the controls. The production cost of MT-APG-NLC (30 ppm) after 21 days of treatment was reduced by 32.9% compared with the conventional MT treatment group (60 ppm). In all the treatments, the length-weight relationship (LWR) showed negatively allomeric growth behavior (b < 3), with a relative condition factor (Kn) of more than 1. Therefore, MT-APG-NLC (30 ppm) would seem to be a promising, cost-effective way to reduce the dose of MT used for the masculinization of farmed red tilapia.

Chicken Enterococcus faecalis-induced immunoglobulin Y as a prophylactic and therapeutic agent against streptococcosis in red tilapia (Oreochromis hybrid).

Background and Aim: Streptococcosis is a common bacterial disease in red tilapia, in which Enterococcus faecalis infection has not been widely reported. This study aimed to evaluate the efficacy of pellets that contain chicken E. faecalis-induced immunoglobulin Y (IgY) to treat and prevent streptococcosis in red tilapia. Materials and Methods: We conducted a 28-day study for immunoprophylaxis and immunotherapy, each using four groups with two replications: Healthy control fish (KS), non-IgY pellets (PA and TA), pellets with 25% egg yolk containing E. faecalis-induced IgY (PB and TB), and pellets with 50% egg yolk containing E. faecalis-induced IgY(PC and TC). Indirect enzyme-linked immunosorbent assay was performed on prototype pellets produced with an IgY suspension at 1.63 mg/mL as the standard optical density curve. For the immunoprophylaxis study, pellets of 3% of the average body weight of the experimental fish (0.50 g per fish per day) were given daily until day 14 before the challenge test with E. faecalis (2.1 × 109 Colony-forming unit/mL peroral) on day 15. The data from the observation period on days 15-28 were analyzed. For the immunotherapy study, pellets of 3% of the average body weight (0.50 g per fish per day) were given daily for 21 days (days 8-28) 7 day spost-infection. The data from the immunotherapy study were collected during the observation period on days 8-28. Statistical analysis was performed on non-specific immune variables: Total leukocytes, monocytes, lymphocytes, neutrophils, phagocytic activity, and macrophage capacity; and the semi-quantitative distribution of melanomacrophage centers (MMCs) in the lymphoid organs, such as spleen and liver. Photomacrographic data were analyzed descriptively and qualitatively by comparing the healing process and clinical signs found between experiments in the immunotherapy study. Results: The pellet with 50% egg yolk with an IgY at 2.43 mg/g pellet, 3% of body weight once daily, was the best formula on experimental fish. The administration of this formulation can also increase non-specific immunity and the distribution of MMCs in the spleen and liver with a survival rate of 55% for 14 days of challenge period in the immunoprophylaxis study and 70% for 21 days of therapy period in the immunotherapy study. Conclusion: Immunoglobulin Y can be a prophylactic and therapeutic agent against streptococcal infections caused E. faecalis in red tilapia with an optimum dosage of 2.43 mg/g pellet.

Emergence of colistin resistance and characterization of antimicrobial resistance and virulence factors of Aeromonas hydrophila, Salmonella spp., and Vibrio cholerae isolated from hybrid red tilapia cage culture.

Background: Tilapia is a primary aquaculture fish in Thailand, but little is known about the occurrence of antimicrobial resistance (AMR) in Aeromonas hydrophila, Salmonella spp., and Vibrio cholerae colonizing healthy tilapia intended for human consumption and the co-occurrence of these AMR bacteria in the cultivation water. Methods: This study determined the phenotype and genotype of AMR, extended-spectrum ß-lactamase (ESBL) production, and virulence factors of A. hydrophila, Salmonella spp., and V. cholerae isolated from hybrid red tilapia and cultivation water in Thailand. Standard culture methods such as USFDA's BAM or ISO procedures were used for the original isolation, with all isolates confirmed by biochemical tests, serotyping, and species-specific gene detection based on PCR. Results: A total of 278 isolates consisting of 15 A. hydrophila, 188 Salmonella spp., and 75 V. cholerae isolates were retrieved from a previous study. All isolates of A. hydrophila and Salmonella isolates were resistance to at least one antimicrobial, with 26.7% and 72.3% of the isolates being multidrug resistant (MDR), respectively. All A. hydrophila isolates were resistant to ampicillin (100%), followed by oxytetracycline (26.7%), tetracycline (26.7%), trimethoprim (26.7%), and oxolinic acid (20.0%). The predominant resistance genes in A. hydrophila were mcr-3 (20.0%), followed by 13.3% of isolates having floR, qnrS, sul1, sul2, and dfrA1. Salmonella isolates also exhibited a high prevalence of resistance to ampicillin (79.3%), oxolinic acid (75.5%), oxytetracycline (71.8%), chloramphenicol (62.8%), and florfenicol (55.3%). The most common resistance genes in these Salmonella isolates were qnrS (65.4%), tetA (64.9%), bla TEM (63.8%), and floR (55.9%). All V. cholerae isolates were susceptible to all antimicrobials tested, while the most common resistance gene was sul1 (12.0%). One isolate of A. hydrophila was positive for int1, while all isolates of Salmonella and V. cholerae isolates were negative for integrons and int SXT. None of the bacterial isolates in this study were producing ESBL. The occurrence of mcr-3 (20.0%) in these isolates from tilapia aquaculture may signify a serious occupational and consumer health risk given that colistin is a last resort antimicrobial for treatment of Gram-negative bacteria infections. Conclusions: Findings from this study on AMR bacteria in hybrid red tilapia suggest that aquaculture as practiced in Thailand can select for ubiquitous AMR pathogens, mobile genetic elements, and an emerging reservoir of mcr and colistin-resistant bacteria. Resistant and pathogenic bacteria, such as resistance to ampicillin and tetracycline, or MDR Salmonella circulating in aquaculture, together highlight the public health concerns and foodborne risks of zoonotic pathogens in humans from cultured freshwater fish.

Effects of Jerusalem Artichoke (Helianthus tuberosus) as a Prebiotic Supplement in the Diet of Red Tilapia (Oreochromis spp.).

The aim of the present study was to evaluate the effects of a Jerusalem artichoke-supplemented diet on the blood chemistry, growth performance, intestinal morphology, expression of antioxidant-related genes, and disease resistance against Aeromonas veronii challenge in juvenile red tilapia. A completely randomized design (CRD) was followed to feed red tilapias with three experimental diets: control, 5.0 g/kg JA-supplemented (JA5), or 10.0 g/kg JA-supplemented (JA10) diets in triplicates for 4 weeks. The results revealed that the growth performance, weight gain (WG), specific growth rate (SGR), and average daily gain (ADG) of fish fed diets JA5 and JA10 were significantly higher (p < 0.05) than those of fish fed the control diet. Fish fed the control diet had significantly higher T-bilirubin, D-bilirubin, and ALT in blood serum than fish fed JA5 and JA10, as well as higher BUN than fish fed JA5. The number of goblet cells in the proximal and distal parts of the intestine revealed that the number of acid, neutral, and double-staining mucous cells of fish fed diets JA5 and JA10 was significantly higher (p < 0.05) than in fish fed the control diet. The diets including the prebiotic (JA5 and JA10) were associated with a significant increase in the expression of gpx1 and gst antioxidant-related genes and disease resistance against A. veronii in juvenile red tilapia. Therefore, JA5 and JA10 can be employed as promising prebiotics for sustainable red tilapia farming.

Effects of Chromium-L-Methionine in Combination with a Zinc Amino Acid Complex or Selenomethionine on Growth Performance, Intestinal Morphology, and Antioxidative Enzymes in Red Tilapia Oreochromis spp.

To consider diet optimization for the growth and health of fish under intensive aquaculture systems, with a focus on the farming of Nile tilapia and red tilapia in Thailand, we conducted an experiment based on a completely randomized design (CRD), with three treatments and four replicates. Three diets, supplemented with different trace minerals, were applied to selected groups of fish: (a) a control diet, without organic trace minerals supplementation; (b) a T1 diet of chromium-L-methionine at 500 ppb, in combination with a zinc amino acid complex at 60 parts per million (ppm); and (c) a T2 diet of chromium-L-methionine at 500 ppb in combination with selenomethionine at 300 ppb. Red tilapia with an initial mean weight of 190 ± 12 g/fish were randomly distributed into cages of 2 × 2 × 2.5 m in a freshwater pond (12 cages in total), with 34 fish per cage and a density of 17 fish/m3. During the 8 week feeding trial, the fish were fed 3−4% of their body weight twice a day. The fish were weighed, then blood samples were collected to study their immune responses. The intestines were collected, measured, and analyzed at the end of the feeding trial. The results showed that the red tilapia that were fed with diets of chromium-L-methionine in combination with a zinc amino acid complex in the T1 treatment had significantly (p < 0.05) higher final weights, weight gains, average daily gains (ADGs), and better feed conversion ratios (FCRs), compared with fish that were fed with the control diet without organic trace minerals and with fish that were fed with the T2 diet (p < 0.05). The midgut and hindgut villus heights of the group fed with chromium-L-methionine in combination with a zinc amino acid complex in the T1 treatment were significantly higher than those of the other groups (p < 0.05). The levels of the antioxidative enzyme superoxide dismutase (SOD) and lysozyme activity were not significantly different from those of fish that were fed with the control diet (p > 0.05), whereas the glutathione level tended to increase (p < 0.1) in fish that were fed with chromium-L-methionine in combination with selenomethionine in the T2 treatment. Therefore, we concluded that chromium-L-methionine in combination with a zinc amino acid complex or selenomethionine clearly enhanced red tilapia's growth performance and feed utilization through the promotion of antioxidative enzyme activity and immune response.

Effect of Treating Eggs with Coenzyme Q10 (CoQ10) on Growth Variables, Histomorphometry, and Antioxidant Capacity in Red Tilapia (Oreochromis aureus × Oreochromis mossambicus) Larvae.

Red tilapia eggs one day post fertilization (dpf) were exposed to coenzyme Q10 (CoQ10) at rates of 0, 5, and 10 mg/L for control, treatment 2 (C5), and treatment 3 (C10), respectively, without exchanging water and until the larval mouth-opening stage. Fertilized eggs of red tilapia exposed to different concentrations of CoQ10 were hatched at rates (p > 0.05) between 38 to 54.67%. The yolk-sac diameter at the 2nd day post hatching (dph), ranged from 1.85 to 1.87 mm in depth and 1.63 to 1.88 mm in width and was not altered by the CoQ10 treatments. Similarly, red tilapia survival (p > 0.05) ranged from 22.67 to 32%. On 6 dph, a slight percentage (2.08%) of survived fishes exposed to high CoQ10 dose (C10) exhibited larval deformation in the form of an axial curvature of the spine in the abdominal and caudal region. Larvae displayed a normal structure of the esophagus folds in all fish groups, and larvae in the C5 group displayed the longest folds and widest muscularis layer, followed by fishes in the C10 group and the control. Red tilapia fry on 30 dph treated with CoQ10 possessed higher antioxidant potentials in terms of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) compared to the control. In conclusion, treating Red tilapia fertile eggs with 5 mg/L CoQ10 improves the growth, gut structure, and antioxidant efficiency of the produced larvae.

Effect of an Oral Bivalent Vaccine on Immune Response and Immune Gene Profiling in Vaccinated Red Tilapia (Oreochromis spp.) during Infections with Streptococcus iniae and Aeromonas hydrophila.

Streptococcosis and aeromonasis inflicted by Streptococcus iniae and Aeromonas hydrophila, respectively, have affected tilapia industries worldwide. In this study, we investigated antibody responses and explored the mechanisms of protection rendered by an oral bivalent vaccine in red tilapia following challenges with S. iniae and A. hydrophila. The results of specific IgM antibody response revealed that the IgM titers against S. iniae and A. hydrophila in the bivalent incorporated (BI) vaccine group were significantly higher (p < 0.05) than those in the bivalent spray (BS) vaccine fish and unvaccinated control fish throughout the experiment. Real-time qPCR results also showed that the gene expression of CD4, MHC-I, MHC-II, IgT, C-type lysozyme, IL-1ß, TNF-α, and TGF-ß remained significantly higher (p < 0.05) than that of the controls between 24 and 72 h post-infection (hpi) in both mucosal (hindgut) and systemic (spleen and head−kidney) organs of BI vaccinated fish. Furthermore, the highest relative expression of the TGF-ß, C-type lysozyme, and IgT genes in the BI vaccinated group was observed in the challenged fish's spleen (8.8-fold), head kidney (4.4-fold), and hindgut (19.7-fold) tissues, respectively. The present study suggests that the bivalent incorporated (BI) vaccine could effectively improve the immune function and activate both humoral and cell-mediated immunities in vaccinated red tilapia following the bacterial challenges.

Effects of resveratrol on lipid metabolism in liver of red tilapia Oreochromis niloticus.

Resveratrol (RES), as a polyphenol natural plant extract, mainly accumulates in the root of Polygonum cuspidatum, which can alleviate liver injury in mammals. Our study aims to explore the effects and potential mechanism of RES on lipid metabolism of red tilapia, and the effects of RES on liver structure, fat synthesis and metabolism of red tilapia were determined. The present study designed four groups named as 8 % fat (8%CK), 10 % fat (10 % HF), 10 % HF + RES and 10 % HF + RES + EX527 (selisistat). The liver tissues of red tilapia were collected at 3 (3 W), 6 (6 W) and 9 (9 W) weeks for parameter determination. Compared to the normal diet group, the hepatocyte of tilapia showed nuclear shift and vacuoles of different sizes when fed a high-fat diet. Meanwhile, the high-fat diet increased the contents of LDL, TC and TG significantly at 6 W, and significantly decreased the content of NAD+ at 9 W. Compared to the high-fat group, the nuclei of tilapia fed with RES were increased and visible, the degree of steatosis and the number of vacuoles were both reduced. At 3/6/9 W, RES significantly decreased the contents of LDL, TG and TMAO, and significantly increased the content of NAD+. A total of 1416 genes were up-regulated and 1928 genes were down-regulated in the group with added RES when compared to the 10 % HF group. The pathways related to lipid metabolism including PPAR signaling pathway have been enriched. Interestingly, the expressions of sirt1, pparα, fabp7 and cpt1b genes were up-regulated in RES diet group, while the expressions of pparγ, me1, scd and lpl genes were down-regulated. After the addition of an inhibitor (EX527), the above indexes showed an opposite trend when compared to the group with added RES. The overall results showed that the high-fat diet could cause fatty liver lesions in the liver of red tilapia, and RES could activate the sirt1 gene, regulate the PPARα/γ pathway and related genes, and thus regulate liver fat synthesis and metabolism leading to the alleviation of damage to liver tissue.

Molecular and functional analysis of the microphthalmia-associated transcription factor (mitf) gene duplicates in red tilapia.

In vertebrates, the microphthalmia-associated transcription factor (mitf) is at the hub of the melanin synthesis regulation network. However, little information is known about its molecular characterization, expression, location, or function in skin color differentiation and variation of red tilapia. The full-length cDNA sequences (1977 bp and 1999 bp) of mitfa and mitfb, encoding polypeptides of 491 and 514 amino acids, were effectively identified from red tilapia in this study. The Mitfa and Mitfb sequences of red tilapia clustered first with O. aureus, then with other teleost fish, according to phylogenetic analysis. Mitfa and mitfb mRNA were highly expressed in the brain, dorsal skin and eye tissues using quantitative real-time PCR. The mRNA expressions of mitfa and mitfb were the highest in the cleavage stage during the early development of red tilapia. Among three different colors of red tilapia, the expression levels of mitfa and mitfb were highest in the PB (pink with scattered black spots) dorsal skin. After overwintering, the mitfa and mitfb mRNA expressions were high in the dorsal skin of PB (color changed from pink to black). Mitfa and mitfb were mostly found in the epidermal layer of the dorsal skin, according to in situ hybridization (ISH) analysis. After injecting mitf-dsRNA duplicates along the tail vein of red tilapia, the activity of tyrosinase and the level of melanin in the dorsal skin both decreased significantly. The mRNA expressions of mitfa and its downstream genes (tyrb, tyrp1a and dct) decreased, whereas the mRNA expression of mitfb increased after mitfa-dsRNA injection. The mRNA expressions of mitfb, tyrb, tyrp1a and dct decreased, whereas the mRNA expression of mitfa increased after injecting mitfb-dsRNA. These findings suggest that mitf gene duplicates may play an important role in red tilapia skin color differentiation and variation via the melanogenesis pathway.

Quality of red tilapia viscera oil (Oreochromis sp.) as a function of extraction methods.

This study aimed to propose a simple and efficient heating-freezing method for oil recovery from red tilapia (Oreochromis sp.) viscera, suitable for industrial application and that does not affect its composition. Three methodologies for oil extraction were studied: a) direct heating (69 °C and 29 min) of samples followed by separation of the oil by decantation, b) direct heating with subsequent freezing and c) solvent extraction assisted by ultrasound. For the oil obtained by each methodology, the following factors were determined: peroxide and iodine values, oxidative stability index, yield percentages and fatty acid profile and, to evaluate the changes thereof, a thermal analysis by differential scanning calorimetry was performed. An oil extracted by centrifugation from fresh viscera was used as control. Results showed yields of 92,126%, 60,99% and 55,36% for the oil obtained by heating and freezing, heating and decanting and solvent extraction, respectively, the other evaluated parameters were similar among each other. The content of PUFA was not affected by heating when compared to the control oil, although a decrease was observed in the solvent extracted oil. This behavior was corroborated with the thermal analysis, which showed that the higher PUFA content, the lower the melting temperatures of the oils and the energy required for phase change. A principal component analysis allowed determining that while there are no differences in the abundance of fatty acids C20:1, 14:0, 18:0, 16:1 and C16:0, there are differences for fatty acids C18:1 and C18:2 depending on the method of extraction used in the oil obtention. The results of this study show that the heating-freezing extraction method is a good alternative for acquiring value-added products and facilitates their implementation in rural areas. Furthermore, allows obtaining a product with high content of polyunsaturated fatty acids (at least a third of the total content).

Transcriptome analysis of skin color variation during and after overwintering of Malaysian red tilapia.

The commercial value of red tilapia is hampered by variations in skin color during overwintering. In this study, three types of skin of red tilapia, including the skin remained pink color during and after overwintering (P), the skin changed from pink color to black color during overwintering and remained black color after overwintering (P-B), and the skin changed from pink color to black color during overwintering but recovered to pink color when the temperature rose after overwintering (P-B-P), were used to analyze their molecular mechanisms of color variation. The transcriptome results revealed that the P, P-B, and P-B-P libraries had 43, 42, and 43 million clean reads, respectively. The top 10 abundance mRNAs and specific mRNAs (specificity measure SPM > 0.9) were screened. After comparing intergroup gene expression levels, there were 2528, 1924, and 1939 differentially expressed genes (DEGs) between P-B-P and P-B, P-B-P and P, and P-B and P, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses of color-related mRNAs showed that a number of DEGs, including tyrp1, tyr, pmel, mitf, mc1r, asip, tat, hpdb, and foxd3, might play a potential role in pigmentation. Additionally, the co-expression patterns of genes were detected within the pigment-related pathways by the PPI network from P-B vs. P group. Furthermore, DEGs from the apoptosis and autophagy pathways, such as baxα, beclin1, and atg7, might be involved in the fading of red tilapia melanocytes. The findings will aid in understanding the molecular mechanism underlying skin color variation in red tilapia during and after overwintering as well as lay a foundation for future research aimed at improving red tilapia skin color characteristics.

Evaluation of chemical silage on egg quality parameters in ISA Brown line laying hens (Gallus gallus domesticus) / Evaluación del ensilado químico sobre los parámetros de calidad del huevo en gallinas ponedoras de la línea ISA Brown (Gallus gallus domesticus)

ABSTRACT In this study we evaluated the inclusion of chemical silage from red tilapia viscera (Oreochromis sp.) in diets for ISA Brown line laying hens (Gallus gallus domesticus) and its influence on nutritional parameters and egg quality. A total of 56 16-week-old laying hens were randomly divided into two groups (one per diet), which in turn were subdivided into 7 groups, each with 4 birds. Eggs were collected during the first 13 weeks of laying. During this period the egg quality parameters were evaluated until week 11; the bromatological characterization of the egg was carried out in the 2nd and 4th week of laying. The results indicated that the inclusion of chemical silage in 17,18% dry matter does not result in statistically significant differences in egg quality parameters or in their bromatological composition, with respect to the control. This led to the conclusion that chemical silage can be used as an alternative to protein inputs such as fish meal and soybean cake in the diet of laying hens, without modifying the quality of the final product.

RESUMEN En este trabajo se evaluó la inclusion de ensilado químico de vísceras de tilapia roja (Oreochromis sp.) en dietas para gallinas ponedoras (Gallus gallus domesticus) de la raza Isa-Brown, y su influencia sobre parámetros nutricionales y la calidad del huevo. Se utilizó un total de 56 gallinas ponedoras de 16 semanas de edad divididas al azar en dos grupos (uno por dieta), los cuales a su vez se subdividieron en 7 grupos de 4 aves cada uno. Se les recolectó huevos durante las primeras 13 semanas de postura. Durante este periodo se realizó la evaluación de los parámetros de calidad del huevo hasta la semana 11. La caracterización bromatológica del huevo se realizo en las semanas 2 y 4, luego de haber sido puestos. Los resultados indicaron que la inclusion de ensilado químico en un 17,18% materia seca, no genera diferencias estadísticamente significativas (p < 0,05) en los parámetros de calidad del huevo ni en su composición bromatológica, con respecto al control. Esto permitió concluir que el ensilado químico puede ser usado como alternativa a insumos proteicos como harina de pescado y torta de soya en la alimentación de gallinas ponedoras, sin modificar la calidad del producto final.

First identification of two co-existing genome-wide significant sex quantitative trait loci (QTL) in red tilapia using integrative QTL mapping.

Red tilapia ( Oreochromis spp .) is one of the most popular fish in China due to its bright red appearance, fast growth rate, and strong adaptability. Understanding the sex determination mechanisms is of vital importance for the selection of all-male lines to increase aquacultural production of red tilapia. In this research, the genetic architecture for sex from four mapping populations ( n=1 090) of red tilapia was analyzed by quantitative trait loci (QTL)-seq, linkage-based QTL mapping, and linkage disequilibrium (LD)-based genome-wide association studies. Two genome-wide significant QTL intervals associated with sex were identified on ChrLG1 (22.4-23.9 Mb) and ChrLG23 (32.0-35.9 Mb), respectively. The QTL on ChrLG1 was detected in family 1 (FAM1), FAM2, and FAM4, and the other QTL on ChrLG23 was detected in FAM3 and FAM4. Four microsatellite markers located within the QTL were successfully developed for marker-assisted selection. Interestingly, three ( lpp, sox14, and amh) of the 12 candidate genes located near or on the two QTL intervals were abundantly expressed in males, while the remaining genes were more highly expressed in females. Seven genes ( scly, ube3a, lpp, gpr17, oca2, cog4, and atp10a) were significantly differentially expressed between the male and female groups. Furthermore, LD block analysis suggested that a cluster of genes on ChrLG23 may participate in regulating sex development in red tilapia. Our study provides important information on the genetic architecture of sex in red tilapia and should facilitate further exploration of sex determination mechanisms in this species.

Immersion Vaccination by a Biomimetic-Mucoadhesive Nanovaccine Induces Humoral Immune Response of Red Tilapia (Oreochromis sp.) against Flavobacterium columnare Challenge.

Immersion vaccination with a biomimetic mucoadhesive nanovaccine has been shown to induce a strong mucosal immune response against columnaris disease, a serious bacterial disease in farmed red tilapia caused by Flavobacterium columnare. However, the induction of a systemic immune response by the vaccine is yet to be investigated. Here, we examine if a specific humoral immune response is stimulated in tilapia by a biomimetic-mucoadhesive nanovaccine against Flavobacterium columnare using an indirect-enzyme-linked immunosorbent assay (ELISA), serum bactericidal activity (SBA) and the expression of immune-related genes within the head-kidney and spleen, together with assessing the relative percent survival of vaccinated fish after experimentally infecting them with F. columnare. The anti-IgM antibody titer of fish at 14 and 21 days post-vaccination was significantly higher in chitosan complex nanoemulsion (CS-NE) vaccinated fish compared to fish vaccinated with the formalin-killed vaccine or control fish, supporting the serum bactericidal activity results at these time points. The cumulative mortality of the unvaccinated control fish was 87% after challenging fish with the pathogen, while the cumulative mortality of the CS-NE vaccinated group was 24%, which was significantly lower than the formalin-killed vaccinated and control fish. There was a significant upregulation of IgM, IgT, TNF α, and IL1-ß genes in the spleen and kidney of vaccinated fish. Significant upregulation of IgM and IgT genes was observed in the spleen of CS-NE vaccinated fish. The study confirmed the charged-chitosan-based mucoadhesive nanovaccine to be an effective platform for immersion vaccination of tilapia, with fish generating a humoral systemic immune response against columnaris disease in vaccinated fish.