Plasma sex steroid hormonal profile and gonad histology were correlated to study the annual reproductive cycle of Hemibagrus nemurus. Hormones were measured by Enzyme Linked Immunosorbent Assay. Gonad tissues were observed by using light microscopy. The highest testosterone (T) value for male was observed in November and that of female was in October. 11-ketotestosterone (11-KT) and 17β-estradiol (E2) levels were highest in June and November, respectively. Hormonal profiles of T, 11-KT and E2 showed several peaks which indicated a non-seasonal pattern. There were significant differences (p
Copper is one of the major heavy metal pollutants found in the aquatic environment. Therefore, it is important for determining the genes that play a key role in copper metabolism in aquatic organisms. This study, thus, aimed to identify a new copper-inducible gene in swordtail fish, Xiphophorus helleri. Using ACP-based RT-PCR coupled with RLM-RACE, we cloned Wap65, a mammalian homologue of hemopexin gene. The gene exhibits high identity at amino acid levels with the Wap65 gene of other fish species (42-68%) and mammalian hemopexin gene (35-37%). In addition, ten cysteine and two histidine residues are conserved in the swordtail fish Wap65 gene. These cysteine residues are vital for structural integrity, and histidine residues provide high binding affinity towards heme. As revealed by RT-PCR, the gene was upregulated in swordtail fish that were exposed to copper in a dose- and time-dependent manner. Therefore, the identification of Wap65, a mammalian homologue of hemopexin, as a new copper-inducible gene will provide greater insight into the role of this gene in copper metabolism.
Dietary organic acids are increasingly being investigated as a potential means of improving growth and nutrient utilization in aquatic animals. A 9-week study was performed to compare equal amounts (2%) of different organic acids (sodium butyrate, acetate, propionate, or formate) on the growth, muscle proximate composition, fatty acid composition, cholesterol and lipid peroxidation, differential cell counts, plasma biochemistry, intestinal short-chain fatty acid (SCFA) level, and liver histopathology to red hybrid tilapia (Oreochromis sp.) (initial mean weight of 2.87 g). A second experiment was performed to determine their effects on lipid peroxidation and trimethylamine (TMA) when added at 1% to tilapia meat and left out for 24 h. The results of the first experiment showed no treatment effect to growth, feeding efficiencies, or muscle fatty acid composition, but all dietary organic acids significantly decreased intestinal SCFA. Dietary butyrate and propionate significantly decreased muscle lipid peroxidation compared to the control group, but the dietary formate treatment had the lowest lipid peroxidation compared to all treatments. Muscle crude protein and lipid in tilapia fed the formate diet were significantly lower and higher, respectively, and showed evidence of stress based on the differential cell counts, significantly higher plasma glucose and liver glycogen, as well as inflammatory responses in the liver. Although a potential benefit of dietary organic acids was a reduction to lipid peroxidation, this could be accomplished post-harvest by direct additions to the meat. In addition, inclusions of butyrate and propionate to tilapia meat significantly decreased TMA, which might be a more cost-effective option to improve the shelf life of tilapia products.
A study was conducted to isolate, partial characterize Asian sea bass (Lates calcarifer) vitellogenin (vtg). Two-year-old juvenile L. calcarifer (n = 10) were given three intraperitoneal injections of 17-β estradiol (E2) at a dose of 2 mg/kg body weight to induce vitellogenesis. Blood was collected 3 days after the last injection, and plasma was purified through gel filtration chromatography. A broad single symmetrical peak consisting of vtg molecule was produced. Protein concentration was 0.059 mg/ml as determined by Bradfrod assay using bovine serum albumin as a standard. The protein appeared as one circulating form in Native PAGE considering the dimeric form of putative vtg with molecular weight of 545 kDa. In SDS-PAGE under reducing conditions, two major bands appeared at 232.86 and 118.80 kDa and minor bands at 100.60, 85.80 and 39.92 kDa, respectively. The purified vtg was used to generate a polyclonal antibody, and the specificity of antibody was assessed by Western blot analysis. Two major bands were immunoreacted, but no cross-reactivity was observed with plasma from non-induced males. The protein was characterized as phosphoglycolipoprotein as it positively stained for the presence of lipid, phosphorus and carbohydrate using Sudan Black B, methyl green and periodic acid/Schiff reagent solution, respectively. The amino acid composition was analyzed by high sensitivity amino acid analysis that showed high percentage of non-polar amino acids (~48 %). The results suggest the potential utilization of vtg as a basis tool to further study about reproductive physiology of this important economical species.
Fish are a major source of beneficial n-3 LC-PUFA in human diet, and there is considerable interest to elucidate the mechanism and regulatory aspects of LC-PUFA biosynthesis in farmed species. Long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis involves the activities of two groups of enzymes, the fatty acyl desaturase (Fads) and elongase of very long-chain fatty acid (Elovl). The promoters of elovl5 elongase, which catalyses the rate-limiting reaction of elongating polyunsaturated fatty acid (PUFA), have been previously described and characterized from several marine and diadromous teleost species. We report here the cloning and characterization of elovl5 promoter from two freshwater fish species, the carnivorous snakehead fish (Channa striata) and zebrafish. Results show the presence of sterol-responsive elements (SRE) in the core regulatory region of both promoters, suggesting the importance of sterol regulatory element-binding protein (Srebp) in the regulation of elovl5 for both species. Mutagenesis luciferase and electrophoretic mobility shift assays further validate the role of SRE for basal transcriptional activation. In addition, several Sp1-binding sites located in close proximity with SRE were present in the snakehead promoter, with one having a potential synergy with SRE in the regulation of elovl5 expression. The core zebrafish elovl5 promoter fragment also directed in vivo expression in the yolk syncytial layer of developing zebrafish embryos.
A 60-day feeding trial was conducted to determine the effects of palm oil blended with oxidized and non-oxidized fish oil on growth performances, hematology, and non-specific immune response in juvenile Japanese sea bass, Lateolabrax japonicas. Japanese sea bass (1.73 ± 0.01 g) were fed seven experimental diets containing 100 g/kg of dietary lipid in forms of palm oil (10P), fish oil (10F), fish oil blended with palm oil at different ratios, 6:4 (6F4P) and 4:6 (4F6P), oxidized fish oil (10OF), and oxidized fish oil blended with palm oil at different ratios, 6:4 (6OF4P) and 4:6 (4OF6P). After the feeding trial, the following results were illustrated. No significant effects were observed in survival, feed conversion ratio, condition factor, and hematocrit after feeding with experimental diets for 60 days. The relatively higher specific growth rate and hematology were observed in 6F4P. Furthermore, both palm oil and oxidized fish oil acted as a negatively on serum lysozyme activity (P < 0.05). This study suggested that a ration of 6F4P is recommended as an innocuous ratio for Japanese sea bass. Furthermore, according to the present investigation, palm oil seems to have the ability to improve the protein efficiency when added to oxidized fish diets as well as a positive trend to the growth performance (P > 0.05).
This study reports the antioxidant property and molecular mechanism of a tryptophan-tagged peptide derived from a teleost fish Channa striatus of serine threonine-protein kinase (STPK). The peptide was tagged with tryptophan to enhance the antioxidant property of STPK and named as IW13. The antioxidant activity of IW13 peptide was investigated using in vitro methods such as DPPH, ABTS, superoxide anion radical scavenging and hydrogen peroxide scavenging assay. Furthermore, to investigate the toxicity and dose response of IW13 peptide on antioxidant defence in vitro, L6 myotubes were induced with generic oxidative stress due to exposure of hydrogen peroxide (H2O2). IW13 peptide exposure was found to be non-cytotoxic to L6 cells in the tested concentration (10, 20, 30, 40 and 50 μM). Also, the pre-treatment of IW13 peptide decreased the lipid peroxidation level and increased glutathione enzyme activity. IW13 peptide treatment upregulated the antioxidant enzyme genes: GPx (glutathione peroxidase), GST (glutathione S transferase) and GCS (glutamine cysteine synthase), in vitro in L6 myotubes and in vivo in zebrafish larvae against the H2O2-induced oxidative stress. The results demonstrated that IW13 renders protection against the H2O2-induced oxidative stress through a cellular antioxidant defence mechanism by upregulating the gene expression, thus enhancing the antioxidant activity in the cellular or organismal level. The findings exhibited that the tryptophan-tagged IW13 peptide from STPK of C. striatus could be a promising candidate for the treatment of oxidative stress-associated diseases.
Despite the potential of vegetable oils as aquafeed ingredients, a major drawback associated with their utilization is the inferior level of beneficial n-3 long-chain polyunsaturated fatty acids (LC-PUFA). Echium oil (EO), which is rich in stearidonic acid (SDA, 18:4n-3), could potentially improve the deposition of n-3 LC-PUFA as the biosynthesis of LC-PUFA is enhanced through bypassing the rate-limiting ∆6 desaturation step. We report for the first time an attempt to investigate whether the presence of a desaturase (Fads2) capable of ∆4 desaturation activities and an elongase (Elovl5) will leverage the provision of dietary SDA to produce a higher rate of LC-PUFA bioconversion. Experimental diets were designed containing fish oil (FO), EO or linseed oil (LO) (100FO, 100EO, 100LO), and diets which comprised equal mixtures of the designated oils (50EOFO and 50EOLO) were evaluated in a 12-week feeding trial involving striped snakeheads (Channa striata). There was no significant difference in growth and feed conversion efficiency. The hepatic fatty acid composition and higher expression of fads2 and elovl5 genes in fish fed EO-based diets indicate the utilization of dietary SDA for LC-PUFA biosynthesis. Collectively, this resulted in a higher deposition of muscle eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) compared to LO-based diets. Dietary EO improved the ratio of n-3 LC-PUFA to n-6 LC-PUFA in fish muscle, which is desirable for human populations with excessive consumption of n-6 PUFA. This study validates the contribution of SDA in improving the content of n-3 LC-PUFA and the ratio of EPA to arachidonic acid (ARA, 20:4n-6) in a freshwater carnivorous species.
To assess the effect of dietary ascorbate on lipid metabolism, 1-year black sea bream (Acanthopagrus schlegelii) were reared on a casein-based purified diet and an ascorbate fortified diet (1,100 mg of L: -ascorbyl-2- monophosphate-Mg/kg diet). The fortified ascorbate was effectively incorporated into the fish body and elevated muscle carnitine content. Fortifications of dietary ascorbate depressed activities of glucose-6-phosphate dehydrogenase and NADP-isocitrate dehydrogenase as lipogenic enzymes in the hepatopancreas and intraperitoneal fat body. Starvation after feeding experiment activated carnitine palmitoyltransferase as a lipolysis enzyme in the hepatopancreas in both control and vitamin C(VC) groups, while the lipolysis activity was significantly higher in VC group. These results confirmed that dietary ascorbate depressed lipogenesis and activated lipolysis, i.e., influenced the lipid metabolism of black sea bream.
A 90-day feeding trial was conducted on the growth performance, feeding efficacy, body indices, various hematological and plasma biochemical parameters, and histopathological examination of the gonads from male and female Nile tilapia fingerlings when fed different crude plant extracts from Cinnamomum camphora, Euphorbia hirta, Azadirachta indica, or Carica papaya at 2 g kg(-1) compared to a control diet. This was followed by a 14-day challenge to Streptococcus agalactiae. All treatments were triplicated, and each treatment consisted of 30 fish. Results showed that C. papaya extracts were the most effective at delaying gonadal maturation to both male and female tilapia, as well as significantly increasing (P < 0.05) growth performance compared to the control treatment. Similarly, dietary C. camphora and E. hirta extracts also significantly improved growth, while no significant growth effect was detected between the A. indica and control treatments (P > 0.05). Further, crude body lipid was lower in the C. camphora, E. hirta and C. papaya treatments, but was only significantly lower for the E. hirta treatment compared to the control. Meanwhile, none of the hematological or biochemical parameters were significantly affected, although plasma ALT was significantly lower for tilapia fed A. indica compared to the control. After the 14-day bacterial challenge, tilapia fed C. camphora supplementation had significantly higher survival, compared to the control, but was not significantly higher than the other supplemented diets. Results indicate that dietary C. papaya extract can significantly promote growth and delay gonadal maturation to both male and female tilapia, while C. camphora was the most effective prophylactic to S. agalactiae and may be a cost-effective and eco-friendly alternative to antibiotics.
Aquaculture has intensified tremendously with the increasing demand for protein sources as the global population grows. However, this industry is plagued with major challenges such as poor growth performance, the lack of a proper environment, and immune system impairment, thus creating stress for the aquaculture species and risking disease outbreaks. Currently, prophylactics such as antibiotics, vaccines, prebiotics, probiotics, and phytobiotics are utilized to minimize the negative impacts of high-density farming. One of the promising prophylactic agents incorporated in fish feed is resveratrol, a commercial phytophenol derived via the methanol extraction method. Recent studies have revealed many beneficial effects of resveratrol in aquatic animals. Therefore, this review discusses and summarizes the roles of resveratrol in improving growth performance, flesh quality, immune system, antioxidant capacity, disease resistance, stress mitigation, and potential combination with other prophylactic agents for aquatic animals.
Catfish are a highly diverse group of fish that are found in various regions across the globe. The significance of catfish culture extends to various aspects, including food security, economic advancement, preservation of cultural legacy, and ecological stewardship. The catfish industry is presently encountering unprecedented challenges as a consequence of the variability in water temperature caused by climate change. Temperature is a significant abiotic component that regulates and restricts fish physiology throughout their life cycle. The impact of severe temperatures on various species of catfish is dependent upon the magnitude of the stressor and additional influencing factors. This paper presents an analysis of the effects of temperature fluctuations on various aspects of catfish species, including growth and survival, blood parameters, enzymatic and hormone response, oxygen consumption rates, sound generation and hearing skills, nutritional requirements, and other phenotypic attributes. While this review is certainly not exhaustive, it offers a broad synopsis of the ideal temperature ranges that are most favorable for several catfish species. In-depth research to investigate the interacting impacts of severe temperature occurrences in conjunction with other associated environmental stresses on a wider variety of catfish species is crucial in order to further our understanding of how catfish species will respond to the anticipated climate change in the future.
Vitamin E, a potent antioxidant consisting of four isomers each (α, β, γ, δ) of tocopherol (T) and tocotrienol (T3), is found naturally in plant oils at different concentrations. In this study, four semi-purified isonitrogenous and isolipidic (10 %) diets containing canola oil, cold-pressed soybean oil, wheat germ oil, or palm fatty acid distillates (PFAD) as the sole vitamin E source were fed to triplicate groups of red hybrid tilapia (Oreochromis sp.) fingerlings (14.82 ± 0.05 g) for 45 days. Vitamin E concentrations and composition were measured in the muscle, liver, skin, and adipose tissue. Deposition of α-T (53.4-93.1 % of total vitamin E) predominated over deposition of other isomers, except in the liver of fish fed the SBO diet, where α-T and γ-T deposition was in the ratio 40:60. T3 deposition (2.6-29.4 %) was only detected in tissues of fish fed the PFAD diet; adipose tissue was the major storage depot. Fish fed the SBO diet contained significantly more (P
This study aims to examine ionoregulatory parameters during exercise and cortisol elevation in common carp fed different food rations. Fish subjected to two different feeding regimes (0.5 or 3.0% body mass (BM) daily) received no implant or an intraperitoneal cortisol implant (250 mg/kg BM) or sham, and were monitored over a 168-h post-implant (PI) period under resting, low aerobic swimming or exhaustive swimming conditions. Plasma osmolality was maintained at relatively stable levels without much influence of feeding, swimming or cortisol, especially in low feeding groups. Nevertheless, a transient hyponatremia was observed in all low feeding fish implanted with cortisol. The hyponatremia was more pronounced in fish swum to exhaustion but even in this group, Na+ levels returned to control levels as cortisol levels recovered (168 h-PI). Cortisol-implanted fish also had lower plasma Cl- levels, and this loss of plasma Cl- was more prominent in fish fed a high ration during exhaustive swimming (recovered at 168 h-PI). Cortisol stimulated branchial NKA and H+ ATPase activities, especially in high ration fish. In contrast, low ration fish upregulated kidney NKA and H+ ATPase activities when experiencing elevated levels of cortisol. In conclusion, low feeding fish experience an ionoregulatory disturbance in response to cortisol implantation especially when swum to exhaustion in contrast to high feeding fish.
Oxyeleotris marmorata is an ambush predator. It is known for slow growth rate and high market demand. Farming of O. marmorata still remains a challenge. In order to establish a proper feeding practice to stimulate growth, knowledge of its metabolic processes and cost should be examined. Therefore, this study was designed to investigate the diel osmorespiration rhythms of O. marmorata in response to feeding challenge by using an osmorespirometry assay. The results have shown that oxygen consumption rate of the fed fish was approximately 3 times higher than that of the unfed fish in early evening to support specific dynamic action. Digestion and ingestion processes were likely to be completed within 18-20 h in parallel with the ammonia excretion noticeable in early morning. Under resting metabolism, metabolic oxygen consumption was influenced by diel phase, but no effect was noted in ammonia excretion. As a nocturnal species, O. marmorata exhibited standard aerobic metabolic mode under dark phase followed by light phase, with high oxygen consumption rate found in either fed or unfed fish. It can be confirmed that both the diel phase and feeding have a significant interactive impact on oxygen consumption rate, whereas ammonia metabolism is impacted by feeding state. High metabolic rate of O. marmorata supports the nocturnal foraging activity in this fish. This finding suggested that feeding of O. marmorata should be performed during nighttime and water renewal should be conducted during daytime.
Lates calcarifer, commonly known as the Asian sea bass or barramundi, is an interesting species that has great aquaculture potential in Asia including Malaysia and also Australia. We have investigated essential fatty acid metabolism in this species, focusing on the endogenous highly unsaturated fatty acid (HUFA) synthesis pathway using both biochemical and molecular biological approaches. Fatty acyl desaturase (Fad) and elongase (Elovl) cDNAs were cloned and functional characterization identified them as ∆6 Fad and Elovl5 elongase enzymes, respectively. The ∆6 Fad was equally active toward 18:3n-3 and 18:2n-6, and Elovl5 exhibited elongation activity for C18-20 and C20-22 elongation and a trace of C22-24 activity. The tissue profile of gene expression for ∆6 fad and elovl5 genes, showed brain to have the highest expression of both genes compared to all other tissues. The results of tissue fatty acid analysis showed that the brain contained more docosahexaenoic acid (DHA, 22:6n-3) than flesh, liver and intestine. The HUFA synthesis activity in isolated hepatocytes and enterocytes using [1-(14)C]18:3n-3 as substrate was very low with the only desaturated product detected being 18:4n-3. These findings indicate that L. calcarifer display an essential fatty acid pattern similar to other marine fish in that they appear unable to synthesize HUFA from C18 substrates. High expression of ∆6 fad and elovl5 genes in brain may indicate a role for these enzymes in maintaining high DHA levels in neural tissues through conversion of 20:5n-3.
Jatropha seed cake (JSC) is an excellent source of protein but does contain some antinutritional factors (ANF) that can act as toxins and thus negatively affect the growth and health status of fish. While this can limit the use of JSC, detoxified Jatropha protein isolate (DJPI) may be a better option. An 8-week study was performed to evaluate dietary DJPI to common carp Cyprinus carpio. Five iso-nitrogenous diets (crude protein of 38%) were formulated that consisted of a C ontrol (fish meal (FM) based protein), J 50 or J 75 (50 and 75% of FM protein replaced by DJPI), and S 50 or S 75 (50 and 75% of FM protein replaced by soy protein isolate, SPI) and fed to triplicate groups of 75 carp fingerlings (75; av. wt. ± SD; 11.4 ± 0.25 g). The growth, feeding efficiencies, digestibility, plasma biochemistry, and intestinal enzymes were measured. Results showed that growth performance of fish fed the S 75- or DJPI-based diets were not significantly different from those fed the C ontrol diet, while carp fed the S 50 had significantly better growth than the J 75 diet. Fish fed the J 75 diet had significantly lower protein and lipid digestibility as well as significantly lower intestinal amylase and protease activities than all other groups. However, all plant protein-based diets led to significantly higher crude protein, crude lipid, and gross energy in the body of common carp compared to the control treatment. Plasma cholesterol and creatinine significantly decreased in the plant protein fed groups, although plasma triglyceride as well as the red blood cells count, hematocrit, albumin, globulin, total plasma protein, and lysozyme activity were higher in plant protein fed groups compared to FM fed group. White blood cells, hemoglobulin concentration, alkaline phosphatase and alanine transaminase activities, and glucose level in blood did not differ significantly among treatments. The results suggest that the DJPI is non-toxic to carp and can be used to replace FM in the diets of common carp up to 75%, but further research to potentially reduce some inherent ANF within this protein source, such as non-starch polysaccharides, may improve nutrient utilization.
In this study, we report the starvation effect and vibriosis infection on a tropical fish, the tiger grouper (Epinephelus fuscoguttatus). The tiger groupers were infected with Vibrio vulnificus for 21 days. Gas chromatography-mass spectrometry combined with multivariate analysis was used to assess the variation in metabolite profiles of E. fuscoguttatus. Metabolite productions in infected fishes were significantly influenced by fatty acid production. The Omega 9 (ω-9) was abundant under the challenged conditions compared to Omega 3 (ω-3) and Omega 6 (ω-6). A total of six fatty acids from the ω-9 group were detected in high concentration in the infected fishes compared to the control groupers. These metabolites are Oleic acid, Palmitoleic acid, 6,9-Octadecenoic acid, 8,11-Eicosadienoic acid, cis-Erucic acid and 5,8,11-Eicosatrienoic acid. The production of ω-9 differed significantly (p ≤ 0.001) in the challenged samples. The detected ω-9 compounds were quantified based on three different extraction techniques with Supelco 37-component FAME mix (Supelco, USA). The highest concentration of ω-9 groups compared to the other fatty acids detected is 1320.79 mg/4 g and the lowest is 939 mg/4 g in challenged-starved; meanwhile, in challenged-fed, the highest concentration detected is 1220.87 mg/4 g and the lowest is 917.25 mg/4 g. These changes demonstrate that ω-9 can be used as a biomarker of infection in fish.
This study was designed to optimize the culture conditions of juvenile Epinephelus fuscoguttatus (Forsskål, 1775) under laboratory conditions. To this effect, the rate of oxygen consumption was monitored as an index of stress under different temperature, salinity, pH, photoperiod, and urea concentrations. The result obtained after 12 h of exposure suggests the preference of the juvenile E. fuscoguttatus to a temperature range of 15-25 °C and salinity of 30 ppt. Based on this study, temperature was found to be the most lethal as 100% mortality was observed after 6 h in fish exposure to temperatures above the optimal (≥ 30 °C). However, the oxygen consumption rate was similar under the different pH, photoperiod, and urea concentration tested. It was concluded that water temperature was most critical in terms of respiration physiology of the juvenile E. fuscoguttatus given the range and levels of environmental factors tested in this study.
This study was designed to evaluate the tolerance of Clarias gariepinus juveniles to a gradual and abrupt increase in salinity over time. To this effect, C. gariepinus juveniles were exposed to three salinity incremental protocols namely 1 g L-1 day-1, 5 g L-1 day-1, and 10 g L-1 day-1. Changes in the hematological parameters and the gill histology of fish were analyzed to determine the impact of osmotic stress on the health status of the fish and its osmoregulatory ability. The result obtained showed that juveniles of C. gariepinus can tolerate salinity stress up to 14 g L-1. At 15 g L-1 and beyond, all samples died regardless of gradual (i.e., 1 g L-1 day-1 administered for 15 days) or abrupt salinity exposure (i.e., 5 g L-1 day-1 administered for three days and 10 g L-1 day-1 administered for two days). Interestingly, more than 90% of the fish survived a direct 10 g L-1 exposure for 24 h without prior acclimation. The hematological parameters accessed in the fish exposed to 10 g L-1 (either gradually or abruptly) showed a significant increase in the white blood cells and a decrease in the red blood cells, packed cell volume, hemoglobin concentration, and all derived blood parameters. The results of the serum biochemistry show a lower total protein and albumin in the salinity-treated fish compared to the control group. However, the serum glucose and the plasma electrolytes (i.e., K+, Na+, and Cl-) were higher in the former group than in the latter. Aside from the stress response expressed in the blood parameters, severe gill degenerations were seen in the histological micrograph obtained for the salinity-treated fish, while the control had a near-normal gill architecture. It was concluded that C. gariepinus could tolerate salinity exposure of 10 g L-1 day-1 (administered gradually or abruptly) and below without killing the fish within 24 h.