Epinephelus Bruneus Research Articles

Molecular characterization and tissue distribution of ferritin M in kelp grouper, Epinephelus bruneus

This study, reports the identification and analysis of ferritin M chain, from kelp grouper, Epinephelus bruneus (EbFerM); it comprises 1004 base pair (bp), including 528bp open reading frame (ORF) which encodes 176 amino acid (aa) residues; the calculated molecular weight is 20kDa. The 5'-untranslated region (UTR) possesses 476bp proceeded by a putative Iron Regulatory Element (IRE). Pair wise alignments showed that EbFerM shared 94% identity with that of Larimichthys crocea and Sciaenops. It is expressed in abundance in liver, spleen, and kidney when challenged with Vibrio anguillarum, lipopolysaccharide (LPS), or poly I:C.

Styrax japonica supplementation diet enhances the innate immune response in Epinephelus bruneus against bacterial and protozoan infections

Kelp grouper, Epinephelus bruneus, fed for 30days with 0% (control), 0.1%, 1.0%, and 2.0% of Styrax japonica supplementation diets, led to reductions in mortality after being challenged with a bacterium (Vibrio harveyi) and a ciliate protozoan (Uronema marinum). The enriched diets significantly increased the survival rate as compared to the controls. The phagocytic and respiratory activities were significantly increased in kelp groupers given 1.0% and 2.0% enriched diets. The complement activity, lysozyme activity, serum bactericidal activity, and total protein level significantly increased with any enriched diet against the pathogens; however antiprotease activity and myeloperoxidase levels significantly increased only with 1.0% and 2.0% enriched diets while the α2-macroglobulin level was significantly enhanced with 1.0% enriched diet. The study suggests that incorporation of S. japonica at 1.0% and 2.0% level in the diet significantly enhances the immune responses in the kelp grouper E. bruneus against V. harveyi and U. marinum.

Kalopanax pictus as feed additive controls bacterial and parasitic infections in kelp grouper, Epinephelus bruneus

Feeding kelp grouper, Epinephelus bruneus (26.1 ± 1.4), with 0%, 0.1%, 1.0%, and 2.0% Kalopanax pictus extract-supplementation diets, for 30 days reduced mortality on being challenged intraperitoneally 100 μl with bacterium Vibrio alginolyticus (2.1 × 10 7 cfu ml −1) and ciliate parasite Philasterides dicentrarchi (2.3 × 10 7 ciliates ml −1). The red blood cells (RBC), white blood cells (WBC), haemoglobin, haematocrit, lymphocytes, and monocytes levels significantly increased in kelp grouper fed with all doses of K. pictus-supplementation diets and challenged with bacterium and parasite when compared to control. However, the levels of mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), neutrophils, and thrombocytes did not significantly. The phagocytic activity, complement activity, and antiprotease activity did not significantly change in kelp grouper fed with 0.1% K. pictus-supplementation diets and challenged with bacterium and parasite. The respiratory activity, lysozyme activity, bactericidal activity, total protein level, and myeloperoxidase levels significantly increased in kelp grouper fed with all the doses of K. pictus-supplementation diet and challenged with bacterium and parasite. However, α2-macroglobulin level significantly increased with 1.0% diet, but not with 0.1% and 2.0% diets. Therefore this study suggests that 1.0% and 2.0% K. pictus-supplementation diets positively protected and enhanced the immune system in kelp grouper E. bruneus against V. alginolyticus and P. dicentrarchi infection.

Korean mistletoe enriched diet enhances innate immune response in kelp grouper, Epinephelus bruneus against Philasterides dicentrarchi

The present study investigated the immunostimulatory effect of Korean mistletoe Viscum album extract (KM-E) on innate immune response in kelp grouper Epinephelus bruneus against Philasterides dicentrarchi. Kelp grouper were divided into four groups of 25 each and fed with 0 (control), 0.5, 1.0, and 2.0% enriched diets with Korean mistletoe extract (KM-E). After feeding for 30 days, the fish were injected intraperitoneally (i.p.) with 100μl of P. dicentrarchi (4.2×107ciliates/ml) to study the immune responses at weeks 1, 2, and 4. The respiratory burst activity did not significantly enhance when fed with 0.5% and 1.0% supplementation diets on week 1 when compared to control diet. On weeks 2 and 4, the respiratory burst activity significantly increased with 1.0% and 2.0% diets. The phagocytic activity significantly enhanced with 1.0% and 2.0% diets, but not with 0.5% diet at any time. When fed with 1.0% KM-E-diet the lysozyme activity did not significantly vary at any week whereas with 1.0% and 2.0% diets it was significantly enhanced. The total protein level significantly increased with 1.0% and 2.0% KM-E-diets from weeks 1 to 4 as compared to control. The present study suggests that 1.0% or 2.0% KM-E-supplementation diet positively enhances the innate immune response in E. bruneus against P. dicentrarchi infection.

Molecular characterization, phylogeny, and expression pattern of c-type lysozyme in kelp grouper, Epinephelus bruneus

Lysozymes are the key molecules in innate immune system and possess high bactericidal properties. In the present study, a full-length c-type lysozyme cDNA has been cloned and characterized by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) techniques from kelp grouper, Epinephelus bruneus (Eb-CTL). The cDNA consists of 753 base pairs (bp) with a 432 bp open reading frame (ORF) that encodes 144 amino acids (aa) residues and polyadenylation signal sequence AATAAA. The deduced aa sequence polypeptide had a predicted molecular weight of 16 kDa and theoretical isoelectric point of 7.3. The deduced aa sequence have a long lysozyme domain which contains all catalytic sites and other conserved residues required for lysozyme activity. Pair-wise alignments showed a higher identity (76.4%) with Psetta maxima lysozyme and low identity (38.9%) with lysozymes of insect Bombyx mori. Interestingly, phylogenetic analysis revealed that the kelp grouper lysozyme was more closely related to other fish and vertebrate lysozymes. Quantitative real-time reverse transcriptase-PCR analysis showed that Eb-CTL transcripts are constitutively expressed in hematopoietic organs such as heart, spleen, and kidney after stimulation with LPS or infection with Vibrio anguillarum, indicating the involvement of Eb-CTL in the innate immunity of kelp grouper. This study is a first step on the genetics and functions of the c-type lysozyme in kelp grouper, and their role in anti-bacterial activity with reference to immunological properties which might have biotechnological applications.

Lactuca indica extract as feed additive enhances immunological parameters and disease resistance in Epinephelus bruneus to Streptococcus iniae

The present study evaluated the efficacy of dietary doses of Lactuca indica extract on immunological parameters and disease resistance against Streptococcus iniae infection in kelp grouper, Epinephelus bruneus. Fishes were fed with L. indica enriched diet at 1.0% and 2.0%; both the diets significantly enhanced NBT level and phagocytic activity on week 2 and 4 compared to control diet whereas the changes did not manifest on first week. The total immunoglobulin significantly enhanced in fish fed with 1.0% and 2.0% enriched diets whereas the lysozyme activity significantly enhanced all the diets from weeks 1 to 4. Further the enriched diets at 1% and 2% level resulted in lower mortality of 45% and 40% respectively indicating higher protection from S. iniae infection than 0.1% diet that resulted in 55% mortality. The results suggest that the dietary supplementation of L. indica extract stimulates the immunological parameters and increases disease resistance in E. bruneus against S. iniae infection.

Influence of diet enriched with green tea on innate humoral and cellular immune response of kelp grouper ( Epinephelus bruneus) to Vibrio carchariae infection

Effect of diet enriched with green tea at 0, 0.01, 0.1 or 1.0% levels on immune responses such as non-specific humoral (lysozyme, antiprotease and complement) and cellular (myeloperoxidase content, production of reactive oxygen, and nitrogen species) and disease resistance on week 1, 2 or 4 in kelp grouper Epinephelus bruneus challenged with Vibrio carchariae (2.47 × 10 8 CFU ml −1) was quantified. At all doses green tea supplementation significantly enhanced the serum lysozyme activity from weeks 1 to 4. On the other hand, after week 2 the serum hemolytic complement activity, leucocyte myeloperoxidase content and reactive nitrogen species protection significantly increased in groups fed with 0.01 and 0.1% green tea supplementation diets. The serum antiprotease activity significantly increased in group fed with at 1.0% green tea from week 1 to 4. However, all diets except at 0.01% level resulted in a significant decrease in reactive oxygen species protection during the experimental period. Challenged groups fed with green tea enriched diet at 0.01 and 0.1% level had a higher relative percent survival than with 1.0% diet on week 1, 2 or 4. The results suggest that dietary administration of green tea supplementation at a concentration of 0.01 and 0.1% level positively enhances the non-specific humoral and cellular immune responses and disease resistance of kelp grouper E. bruneus to V. carchariae.

Immunomodulatory effect of sodium alginate enriched diet in kelp grouper Epinephelus brneus against Streptococcus iniae

The effect of diets containing sodium alginate at 0, 0.5, 1.0, and 2.0 g kg −1 following challenge with Streptococcus iniae in kelp grouper Epinephelus bruneus were assessed with reference to survival rate and innate immune parameters such as alternative complement, lysozyme, natural haemagglutination, respiratory burst, superoxide dismutase, and phagocytic activities on week 1, 2, and 4. Fish fed with sodium alginate containing diet at 1.0 and 2.0 g kg −1 after being challenged with S. iniae had higher survival rates of 75% and 60%, respectively than those fed with control diet (0 g kg −1). With any enriched diet the percentage of macrophages significantly decreased from week 1–4, while the percentage of neutrophils and lymphocytes significantly increased. The alternate complement activity, natural haemagglutination, and phagocytic activities of infected fish fed with sodium alginate containing diet at 1.0 g kg −1 on week 2 and 1.0 and 2.0 g kg −1 diets on week 4 were significantly higher when compared to the control. The lysozyme, respiratory bursts, and superoxide dismutase activities of fish fed with enriched diets at 1.0 and 2.0 g kg −1 were significantly increased on week 2 and 4. We therefore recommend that at 1.0 or 2.0 g kg −1 dietary administration of sodium alginate can enhance innate immunity and disease resistance in kelp grouper against S. iniae.

Molecular cloning of GnRH1 gene and GTH cDNAs of the protogynous longtooth grouper, Epinephelus bruneus

In teleosts, gonadotropin-releasing hormone (GnRH) and gonadotropin hormone (GTH) play important roles in regulating gonadal development and maturation. In Southeast Asia, the longtooth grouper, Epinephelus bruneus, is a commercially important aquaculture fish. In this study, we cloned and characterized the longtooth grouper GnRH1 gene and cDNAs of three gonadotropin subunits (GTHα, FSHβ, LHβ). The GnRH1 gene of longtooth grouper was 4, 032 bp long, and contained four exons, 61, 151, 99, and 423 bp long. GTHα, FSHβ, and LHβ cDNAs were 509, 576, and 579 bp, respectively. Phylogenetic and Southern hybridization analyses revealed that the longtooth grouper GTH subunits were evolutionarily close to those of groupers and are one-copy genes. RT-PCR analyses showed that GTH subunit mRNAs were expressed at a higher level in the pituitary glands of immature fish than in those of mature fish, suggesting a role in gonadal maturation.

Diet enriched with mushroom Phellinus linteus extract enhances the growth, innate immune response, and disease resistance of kelp grouper, Epinephelus bruneus against vibriosis

The effect of diet supplemented with Phellinus linteus fed for 30 days was investigated in grouper Epinephelus bruneus challenged with Vibrio anguillarum, Vibrio harveyi, Vibrio alginolyticus, and Vibrio carchariae; infected and treated fish had a significantly higher percent weight gain and feed efficiency. In groups fed with enriched diet and challenged with V. anguillarum and V. harveyi the mortality rate declined with a consequent rise in survival rate than with other pathogens. On the other hand, in groups fed with P. linteus enriched diet and challenged with V. anguillarum, V. harveyi, and V. alginolyticus the cellular and humoral immune responses, such as the alternative complement activity (ACH 50), serum lysozyme activity, phagocytic activity (PA), phagocytic index (PI) significantly higher than in the control group. The respiratory bursts (RB), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activities were found significantly enhanced when the groups fed with enriched diet against V. anguillarum and V. harveyi. The results reveal that kelp grouper fed for 30 days with P. linteus enriched diet had higher cellular and humoral immune response and disease protection from vibriosis than the group fed on basal diet with the protection linked to stimulation of immune system.

Lactobacillus sakei BK19 enriched diet enhances the immunity status and disease resistance to streptococcosis infection in kelp grouper, Epinephelus bruneus

The effect of Lactobacillus sakei BK19 (10 8 cells g −1) supplemented diet fed to kelp grouper, Epinephelus bruneus against streptococcosis caused by Streptococcus iniae and Streptococcus parauberis with reference to the innate immune response and disease resistance was evaluated at 1, 2, and 4 weeks. Maximum reduction in mortalities was observed in kelper feeding the probiotic diet for two weeks after challenged with the pathogens when compared to the infected group fed with basal diet; similarly the cellular and humoral immune responses such as head kidney macrophage phagocytic and peroxidase activities, serum lysozyme activity, and total protein levels increased significantly. The results reveal that, in streptococcosis infected kelp grouper feeding L. sakei BK19 enriched diet affords a higher level of disease protection due to stimulation of immune system.

Effect of dietary lipid level on growth performance and feed utilization of juvenile kelp grouper Epinephelus bruneus

A growth trial was conducted to evaluate the effect of dietary lipid level on juvenile kelp grouper Epinephelus bruneus. Juvenile kelp grouper were fed isonitrogenous diets (470 g/kg crude protein) with four levels of crude lipid at 60 g/kg (CL60), 130 g/kg (CL130), 210 g/kg (CL210), and 270 g/kg (CL270) for 56 days. The highest growth performance and feed utilization were found in the CL130 diet group. A high dietary lipid level (CL270 diet) significantly decreased growth performance and feed utilization. A significant difference in apparent digestibility was only observed in protein, which was highest in the CL130 diet groups. The highest retention for protein, energy, and lipid was found in the CL130 diet group. The dietary lipid levels significantly changed whole-body and liver compositions, the highest being the CL60 diet group for crude protein level and the CL210 diet group for crude lipid level. Based on a second-order polynomial regression analysis of crude lipid level against specific growth rate and protein efficiency ratio, the optimum dietary lipid level for kelp grouper was estimated to be 152 and 154 g/kg diet, respectively.

Shrinking baseline: the growth in juvenile fisheries, with the Hong Kong grouper fishery as a case study

AbstractHistoric and current information on the grouper fishery from Hong Kong and adjacent waters reveals significant changes in species composition and fish sizes over the past 50 years in this important Asian centre for seafood consumption. Once dominant, large groupers are now rare and small species and sizes prevail in the present‐day fishery. Juveniles comprise over 80% of marketed fish by number among the most commonly retailed groupers, and reproductive‐sized fish are absent among larger species. Current fishery practices and the lack of management in Hong Kong and adjacent waters pose a significant threat to large species with limited geographic distribution such as Epinephelus akaara and Epinephelus bruneus, both now listed as threatened by the IUCN. The heavy reliance on juveniles, not only for groupers, but for an increasing diversity of desired fishes within Asia, potentially reduces stock spawning potential. The ‘shrinking baseline’ in terms of a progressive reduction in fish sizes being marketed in the region can seriously undermine fishery sustainability and recoverability of depleted fish stocks. Fishing pressure on groupers and other valuable food fishes within the Asia‐Pacific is intensifying, the declining long‐term trend of grouper landings in Hong Kong and the increasing focus on juveniles for immediate sale or for mariculture ‘grow‐out’ signal a worrying direction for regional fisheries. Moreover, the common appearance of small groupers for sale will influence public perception regarding what are ‘normal‐sized’ fish. Management attention is needed if these fisheries are to remain viable.

Long‐term effects of passive integrated transponder (PIT) tagging on the kelp grouper Epinephelus bruneus

The effects of passive integrated transponder (PIT) tagging on the growth and survival of kelp grouper Epinephelus bruneus was examined over 1 year. The overall average mass gain for the 1 year period of the abdominal cavity tagged group was higher than the dorsal muscle tagged group. No significant differences were found in overall specific growth rate and food conversion between control and treatment groups.

Anaesthetic efficacy and physiological responses to clove oil-anaesthetized kelp grouper Epinephelus bruneus

The efficacy of clove oil as an anaesthetic and at producing a physiological response (plasma cortisol and glucose) was evaluated in the kelp grouper, Epinephelus bruneus. To acquire complete anaesthesia in less than 3 min and recovery in <10 min, three doses of clove oil were tested at 18, 22 and 26 °C. Although higher anaesthetic doses resulted in shorter induction times and longer recovery times, and a lower temperature resulted in longer anaesthesia induction and slower recovery, we found the optimal dose and administering temperature of clove oil to be 250–300 mg L−1 at water temperature of 18 °C, 150–200 mg L−1 at water temperature of 22 °C and 50–100 mg L−1 at water temperature of 26 °C respectively. Following the administration of 150 mg L−1 of clove oil at 22 °C, the plasma cortisol level was highest (4.24 ± 1.571 μg dL−1) after 12 h and the plasma glucose was highest (92.7 ± 9.61 mg dL−1) after 2 h. These results should be useful to the aquaculture industry, where anaesthesia is necessary for a range of activities.

Evaluation of Povidone-iodine Treatment for Disinfection and Normal Hatching Rate on Fertilized Eggs of Longtooth Grouper, Epinephelus bruneus

Evaluation of Povidone-iodine Treatment for Disinfection and Normal Hatching Rate on Fertilized Eggs of Longtooth Grouper, Epinephelus bruneus

Larval Development and Rearing of Longtooth Grouper Epinephelus bruneus in Jeju Island, Korea

AbstractEggs and sperm were obtained from a female (6.3 kg/BW) and a male (8.4 kg/BW) longtooth grouper Epinephelus bruneus following HCG injection in July 2003. The eggs were fertilized artMcially with the sperm and incubated in one of two 50‐m3 tanks after washing the fertilized eggs. The fertilized eggs were 830–950 pn (average 900 ± 2 μm) in diameter and the respective fertilization and hatching rates were 97.7 ± 0.6% and 96.8 ± 0.5% at a water temperature of 25.0 ± 0.5 C. With this regime, the survival rate by day 93 was 7.5% in the 50‐m3 tank. The elapsed time from hatching to opening the mouth was 3 d at 25 C. The initial mouth size (z) of the larvae was 0.22–0.23 mm. The newly hatched larvae were 2.02 ± 0.02 mm TL; this increased to 4.12 ± 0.09 mm TL by day 11. By day 54, the larvae had metamorphosed into juveniles and reached 41.12 ± 1.20 mm TL, and by day 93 the juveniles reached 93.78 ± 1.98 mm TL. In all, 49.5% of the larvae were malformed and the type of malformation was diverse.

Ontogeny of digestive and immune system organs of larval and juvenile kelp grouper Epinephelus bruneus reared in the laboratory

: A histological examination was made of the ontogenetic development of the digestive and immune systems of the larval and juvenile kelp grouper Epinephelus bruneus reared in the laboratory. The liver, gall bladder, pancreas and the demarcating region between the intestines and rectum were formed within 3 days post-hatch (dph). During the preflexion phase (within 16 dph), revolution of the intestine concluded, and pharyngeal teeth and the mucous cells of the esophagus were differentiated. In the transitional period to the juvenile stage (25 dph), the blind sac of the stomach, gastric glands and pyloric caeca began to form. From the viewpoint of the differentiation phase of the adult-type digestive system, the kelp grouper is similar to Heterosomata, hitherto reported. The primordial thymus, kidney and spleen were present at 12, 1 and 6 dph, and the small lymphocytes in these lymphoid organs appeared at 21, 30 and 33 dph, respectively. The developmental sequence of the lymphoid organs and the appearance ages of the lymphoid organs and small lymphocytes in the lymphoid organs in the kelp grouper are similar to those of other marine fish previously reported, except for the Pacific bluefin tuna Thunnus orientalis.