Effect of dietary mannan oligosaccharide on growth,body composition,haematology and biochemical parameters of Asian seabass (Lates calcarifer)

A feeding trial was conducted to study the effect of mannan oligosaccharide (MOS) supplementation on growth, body composition, haematology, biochemical parameters and histology of Asian seabass (Lates calcarifer) fingerlings (8.13 ± 0.06 g average weight). Mannan oligosaccharide was supplemented at five different concentrations viz., 0%, 0.5%, 1%, 1.5% and 2% levels in the diet (40% protein and 9% lipid) of L. calcarifer. The results of the 60 days feeding trial showed that there was a significant (P < 0.05) increase in the final body weight, weight gain (WG), WG%, average daily gain, survival, specific growth rate, daily growth coefficient, hepatosomatic index and viscerosomatic index in the fish fed diet containing 1% MOS. Whole body composition of post‐fed animals showed non‐significant differences (P > 0.05) among the various treatment groups. The analysis of haematological parameters showed that there was no significant (P > 0.05) differences among different treatments but for the haemoglobin content which was significantly (P < 0.05) higher in the fish fed 1% MOS. Biochemical parameters revealed that glucose, urea, cholesterol and triglyceride content showed significant (P < 0.05) difference between control and MOS‐supplemented group. Histological observations of post‐fed animals revealed that MOS supplementation resulted in increased absorptive surface area of the intestine and increased glycogen deposition in liver. The result of this experiment infers that MOS supplementation has got a beneficial effect in the diet of seabass and supplementation at 1% level is optimal for improving the growth.     

Vibrio alginolyticus infection in Asian seabass (Lates calcarifer,Bloch) reared in open sea floating cages in India

Asian seabass (Lates calcarifer) has been designated as a candidate species for open sea cage culture in India. A case of vibriosis in Asian seabass reared in open sea floating cages is reported. Haemorrhage and ulcer were observed grossly in the diseased fish. The pathogen, identified as Vibrio alginolyticus based on biochemical and molecular characterization, was isolated from liver, gill, kidney, brain and blood. Histological examination of the diseased fish showed congestion, haemorrhage and necrosis in vital organs. The LD₅₀ studies showed that the organism was virulent to Asian seabass, and the LD₅₀ value was 10^3.2 CFU g^?1 fish.     

Genetic relatedness and differentiation of hatchery populations of Asian seabass (Lates calcarifer) (Bloch, 1790) broodstock in Thailand inferred from microsatellite genetic markers

After more than 20 years of hatchery production of Asian seabass in Thailand, genetic information is still lacking for effective genetic management and a selective breeding programme. This study aimed to evaluate genetic status of existing hatchery populations and genetic consequences of a selective breeding attempt. We examined genetic relatedness in seven hatchery samples, including a selectively bred population (RACF‐F1), compared with three wild samples using 11 microsatellite loci. Genetic diversity and relatedness values within most hatchery samples, except for RACF‐F1, did not differ from those of wild populations (P > 0.05). RACF‐F1 had the lowest allelic diversity and effective population size (A_r = 6.99; N_e = 7.8) and highest relatedness values (mean r_xy = 0.075–0.204). Pairwise Φ_ST values, principal component analysis and model‐based cluster analyses revealed three genetically distinct hatchery groups: Eastern Thailand (CHN, RACF, NSCF and SKCF), Southern Thailand (NICA) and the Andaman Sea (STCF). Results suggest that exiting domestic populations capture reasonable amount of genetic variation and can be useful for a base population for genetic improvement programmes. In addition, given the rapid increase in relatedness that we observed in one selectively bred population, we recommend using selection methods and hatchery practices that reduce variability in family contribution in the subsequent generations.     

Development of two cell culture systems from Asian seabass Lates calcarifer (Bloch)

Two new cell culture systems namely epitheloid cells of Lates (LCE) and fibroblastic cells of Lates (LCF) have been developed from fry and fingerling of the economically important brackishwater fish Lates calcarifer. Primary cultures were initiated by explant technique using caudal fin of fingerling and whole body tissue of the fry. The nutritional requirements and the growth pattern in response to different culture environment were similar for the two cell cultures. The culture medium used was Leibovitz‐15 supplemented with 20% fetal bovine serum (FBS) and 1% fish serum. The LCE comprised of epithelioid cells and LCF cells were fibroblastic. With a split ratio of 1:2, the confluency of cells was attained in 8–10 days at an incubation temperature of 28°C. The cells were found to grow well in a wide range of temperature (24–32°C) and stable at 20 and 36°C. The growth rate of LCF and LCE cells increased proportionately with the concentration of FBS from 5% to 20%. A decrease of serum level to 10% after eight subcultures produced no apparent change in cell morphology and growth rate. The viability of cells was found to be 70% when revived after a month of storage in liquid nitrogen (?196°C).     

Visual thresholds for feeding and optimum light intensity for larval rearing of Asian seabass,Lates calcarifer (Bloch)

This study examined the retinomotor responses and prey ingestion rates of 10‐, 15‐, 20‐ and 30‐day‐old Asian seabass Lates calcarifer under different light intensities from 0 to 1000 lx to determine the visual thresholds. Subsequently, two age groups of seabass larvae were reared under light intensities of 10, 100 and 1000 lx to determine the optimum illumination in hatchery tanks. Retinomotor response was absent in 10‐day‐old larvae, but quite marked in 15‐ and 20‐day‐old seabass at 1 lx and higher. Ingestion of Artemia nauplii by 10‐day‐old larvae was almost zero at <1 lx, increased significantly at 1 lx, and was maximal at 10–100 lx. Artemia ingestion under dim light <1 lx improved with age, and older larvae took more prey in complete darkness due to the presence of rod cells (and also free neuromasts). Larvae from 13 to 26 days group had similar survival and growth at 10–1000 lx, however, from 5 to 10 days group showed similar survival rate with highest weight gain at 100 lx. Therefore, we recommend that hatchery rearing tanks be illuminated such that the larvae in the water are exposed to approximately100 lx.     

Dietary administration of astaxanthin improves feed utilization,growth performance and survival of Asian seabass,Lates calcarifer (Bloch, 1790)

This study was undertaken to assess the impacts of dietary astaxanthin supplementation on growth performance, feed utilization, survival, and serum growth hormone (GH) availability of Asian seabass, Lates calcarifer, with special reference to dose–response relationships and variations during different feeding phases (short‐term, medium‐term and long‐term). Fish were fed the following diets in triplicate for 90 days: the control (CD), AX50 (50 mg astaxanthin/kg diet), AX100 (100 mg astaxanthin/kg diet) and AX150 (150 mg astaxanthin/kg diet). The findings revealed that fish exhibited significant linear increments (p < .05) in specific growth rate (SGR), weight gain, feed utilization efficiency and survival when fed various diets with escalating levels of astaxanthin. Supplementation with dietary astaxanthin significantly augmented (p < .05) GH levels in fish. Significant positive associations (p < .05) were observed between circulating serum GH levels and SGR of fish from all groups following three consecutive feeding phases, denoting a robust cause‐and‐effect relationship. Circulating GH concentrations were considered as a sensitive biomarker of growth performance in Asian seabass. This study illustrated that supplemental astaxanthin could be administered in culture protocols to improve the growth rate and commercial hatchery production of Asian seabass, and possibly other teleost species.     

Production of protein hydrolysates from skipjack tuna (Katsuwonus pelamis) viscera as feeding attractants for Asian seabass (Lates calcarifer)

Skipjack tuna viscera hydrolysate (TVH) was produced by enzymatic hydrolysis using alcalase. A study of the effect of time and enzyme concentration was performed and formed the basis for preparing TVH with the desired degree of hydrolysis (DH). An enzyme to substrate ratio of 5 g kg⁻¹ (TVH A1) and 16.4 g kg⁻¹ (TVH A2) at pH 8.0 with a temperature of 50 °C, and hydrolysis time of 60 min were found to be optimal conditions to obtain hydrolysis of 42.9% and 62.9%, respectively. Both TVH A1 and TVH A2 were analysed for chemical and amino acid composition and peptide size. The effectiveness of TVH at different DH as feeding attractants was estimated in a feeding trial at different inclusion levels. Ten experimental diets , including a fish meal‐based reference diet, were fed to Asian seabass for 28 days. Fish fed the diet supplemented with 30 g kg⁻¹ of TVH A2, and the reference diet showed the highest feed intake (P < 0.05). The growth performance of fish fed diets supplemented with TVH was slightly higher than those of fish fed the control diet, but the reference group showed a significantly higher growth rate (P < 0.05). These findings suggest that the supplementation of feed with TVH that has been processed under suitable conditions can improve feed palatability and growth performance when utilizing alternative protein sources in Asian seabass diets.     

Combined or individual effects of dietary magnesium and selenium nanoparticles on growth performance,immunity, blood biochemistry and antioxidant status of Asian seabass (Lates calcarifer) reared in freshwater

This study was conducted to investigate effects of and interactions between nanoselenium (NanoSe) and nanomagnesium (NanoMg) on growth, humoral immunity, serum biochemistry and antioxidant capacity of juvenile Asian seabass Lates calcarifer reared in freshwater. Four groups of fish with an average weight of 32.78 ± 2.23 g were fed one of the experimental diets for 6 weeks: (a) control (basal diet); (b) NanoSe (basal diet + 4 mg NanoSe/kg diet); (c) NanoMg (basal diet + 500 mg NanoMg/kg diet); and (d) combination (basal diet + 4 mg NanoSe/kg diet + 500 mg NanoMg/kg diet). Fish fed with NanoSe‐supplemented diets (NanoSe and combination) showed higher weight gain, specific growth rate and feed intake. The combination of NanoSe and NanoMg enhanced the immune response. Also, fish fed on combination diet showed higher serum alanine aminotransferase and aspartate transaminase levels but lower glucose concentration. The activities of liver superoxide dismutase, glutathione peroxidase and catalase were not different among all groups, but liver malondialdehyde level was lower in fish fed diets supplemented with NanoSe and/or NanoMg. It could be concluded that NanoSe improved growth performance; the combination of both NanoSe and NanoMg enhanced humoral immunity; and NanoSe and/or NanoMg improved antioxidant capacity in Asian seabass.     

Effect of dietary chitosan on growth performance,haematology, immune response,intestine morphology,intestine microbiota and disease resistance in gibel carp (Carassius auratus gibelio)

A 75‐day experiment was conducted with juvenile gibel carp (Carassius auratus gibelio) (4.80 ± 0.01 g) to evaluate effects of dietary chitosan on fish growth performance, haematology, intestine morphology and immune response. Six isonitrogenous (crude protein: 383 g kg^?1), isolipid (97.5 g kg^?1) and isocaloric (gross energy: 16.7 kJ g^?1) diets were formulated to contain 0, 1800, 4000, 7500, 10 000, 20 000 mg kg^?1 chitosan, respectively. The results showed that the growth was depressed when the fish fed with 10 000 mg kg^?1 chitosan. Serum cholesterol, triglyceride and low‐density lipoprotein decreased in 10 000 and 20 000 mg kg^?1 chitosan. On day 75, blood leucocyte phagocytic activity respiratory burst and alternative pathway of complement haemolytic activity were enhanced in 4000 mg kg^?1 chitosan. The number of goblet cell, intraepithelial lymphocyte of mid‐intestine and microvilli height of distal intestine increased at 4000 mg kg^?1 dietary chitosan. Dietary chitosan modulated intestine microbiota, depressed pathogen bacteria Aeromonas veronii‐like and improved Cellulomonas hominis‐like, Bacillus oceanisediminis‐like and two uncultured bacterium‐like species on day 75. Dietary 7500 and 10 000 mg kg^?1 chitosan enhanced the protection against Aeromonas hydrophila infection. In conclusion, oral administration of dietary 7500 mg kg^?1 chitosan for 75 days is recommended for the survival of gibel carp.     

Dietary administration of natural immunostimulants on growth performance,haematological, biochemical parameters and disease resistance of Asian Sea bass Lates calcarifer (Bloch, 1790)

This study was aimed to address the promising evaluation of Cissus quadrangularis plant (stem) and lipopolysaccharide (LPS from bacteria) supplemented diets on innate immune response in Lates calcarifer fingerlings against Aeromonas hydrophila infection. Fingerlings were fed supplemented diets containing four different concentrations of C. quadrangularis (0.5, 1.0, 1.5 and 2.0 g kg^?1 feed), LPS (25, 50, 75 and 100 mg kg^?1 feed) and control (normal formulated diet) for 60 days. The fish fingerlings fed supplemented diet displayed significant differences (P < 0.05) in specific growth rate (SGR) and relative percentage survival compared to the control group fed without C. quadrangularis and LPS‐supplemented diet. Fingerlings were injected intraperitoneally with 100 μL lethal dose of A. hydrophila containing 1 × 10⁶ CFU g^?1. Supplementation of C. quadrangularis and LPS diet significantly increased biochemical profile such as protein, lipid and carbohydrate content, haematological parameters of L. calcarifer fingerlings in different experimental periods when compared with the control group. Dietary doses of C. quadrangularis and LPS‐supplemented diet significantly influenced growth performance and increased survival rate in L. calcarifer fingerlings against A. hydrophila infection.     

Effect of dietary chitosan oligosaccharide complex with Ce (IV) on growth,immunity and disease resistance against Vibrio splendidus of sea cucumber,Apostichopus japonicas

An 8‐week feeding trial was conducted to investigate the effects of dietary chitosan oligosaccharide complex with cerium (Ce IV) (COS‐Ce) on growth performance, nonspecific immunity and disease resistance of sea cucumber, Apostichopus japonicas. Five isonitrogenous (18.6%) and isolipidic (1.1%) practical diets were formulated with graded level of COS‐Ce (0, 150, 300, 600 and 1200 mg kg^?1 dry feed), which were named as COS‐Ce/0, COS‐Ce/150, COS‐Ce/300, COS‐Ce/600, COS‐Ce/1200 respectively. Each diet was allocated to four replicates of sea cucumbers (Initial weight: 6.72 ± 0.02 g). Sea cucumbers were fed to apparent satiation once daily (19:00 hours) for 56 days. During the experiment, water temperature was kept at 16 ± 0.5°C, pH 7.8–8.2, dissolved oxygen beyond 5 mg L^?1, ammonia nitrogen below 0.5 mg L^?1 and salinity from 30‰ to 31‰. Results showed that the specific growth rate of sea cucumbers was significantly higher in COS‐Ce/600 than that in other four treatments. Activities of phagocytosis, respiratory burst, acid phosphatase and alkaline phosphatase in COS‐Ce/600 were significantly higher than that in COS‐Ce/0 (P < 0.05) respectively. On the contrary, cumulative mortality was the lowest in COS‐Ce/600 following 14 days exposure to Vibrio splendidus (P < 0.05). In conclusion, these results confirmed that dietary COS‐Ce had beneficial effects on growth performance, nonspecific immunity and disease resistance of sea cucumber.     

Influence of fish size and water temperature on the metabolic demand for oxygen by barramundi, Lates calcarifer (Bloch), in freshwater

Oxygen demand by all animals is driven primarily by their needs for sustaining metabolism. Typically, larger animals require more oxygen and cellular fuel to carry out respiration than smaller animals. This relationship in most cases is not linear and is usually described by a coefficient and exponent (e.g. ax^b): the exponent b showing the relationship between live‐weight and energy/oxygen demand and is often termed the metabolic body weight (MBW) exponent, while the coefficient (a) tends to be temperature specific and describes the relationship between MBW and maintenance metabolic energy and oxygen demand at that specific temperature. Across all temperatures (range 26.0–32.0°C), the relationship between barramundi (Lates calcarifer) live‐weight (x; g) and relative oxygen consumption as standard metabolic rate (y; mg O₂ kg^?1 h^?1) at 29.4±1.5°C (mean±SD) was described by the exponential curve: y=710.19 x^?0.3268, R²=0.6875 (n=222 assessments). Examination of the same data but on a gross oxygen consumption (mg O₂ h^?1) basis showed a relationship between live‐weight (x; g) and gross oxygen consumption (y; mg O₂ h^?1) that was described by the exponential curve: y=0.710 x^0.6732, R²=0.9033. Evaluation of the combined relationship between fish live‐weight (y; g) and water temperature (x; °C) on gross oxygen consumption rate (z; mg O₂ h^?1) was described by the equation: z=(?20.7818+1.4017x?0.0227x²) ×y^0.673.     

壳聚糖和益生菌对暗纹东方鲀抗病力和免疫功能的影响

为了解壳聚糖和益生菌刺激和增强免疫应答的可能作用,本试验在基础饲料中添加0.2%、0.5%、1.0%的壳聚糖,0.1%、0.2%、0.4%的益生菌,甘露聚糖与益生菌混合物,壳聚糖与益生菌的混合物,在室内水泥池中喂养暗纹东方鲀(3.15±0.05)g 2个月,根据抗菌活力筛选出免疫增强饲料组(0.2%壳聚糖,0.1%、益生菌,甘露聚糖与益生菌混合物,壳聚糖与益生菌混合物)测定免疫功能。结果表明,甘露聚糖与益生菌混合物仅提高头肾T淋巴细胞转化,壳聚糖与益生菌混合物显著增强头肾T、B淋巴细胞转化和脾脏B淋巴细胞体外培养时分泌的IgM。筛选出的四种免疫增强饲料均能使头肾B淋巴细胞体外培养时分泌的IgM显著增加。无论是脾脏B淋巴细胞还是头肾B淋巴细胞,体外培养时所分泌的IFN-α都不同程度地受抑制。     

Use of neem oil (Azadirachta indica) to control caligid copepod infestation on Asian seabass (Lates calcarifer)

Asian seabass (Lates calcarifer) culture in marine cages have been developed rapidly in Malaysia. The high intensity of culture might facilitate several disease infection including caligid infestation. This study was conducted to evaluate the effect of neem oil (Azadirachta indica) against caligid parasites on seabass. The prevalence and clinical pathogen of Caligus infection has been described through a sampling of 150 fish from floating cage at different cultured periods. Acute toxic tests with different concentrations of neem oil were carried out on the parasite and fish host. Results revealed that the 96‐hr median lethal concentration (LC₅₀) of Caligus and fish (10.3 ± 2.5 g of body weight) were 2 and 20 ppm respectively. The in vivo test indicated that 10 ppm of neem oil could result in 100% antiparasitic efficacy within 96 hr. These results therapeutically demonstrated the efficacy of neem oil in caligid control.     

Dietary influence of Fucoidan supplementation on growth of Lates calcarifer (Bloch)

Sulphated polysaccharides exhibit a range of activities which promote health and growth benefits to aquaculture species. One class of sulphated polysaccharides derived from marine algae is fucoidan, which is suggested to exhibit an affinity to the negative muscle growth inhibitory protein myostatin. Addition of fucoidan to the diet of fish therefore may lead to enhancements in growth through inactivation of the myostatin protein. To examine the growth enhancing potential of this sulphated polysaccharide, fucoidan extracts from Undaria pinnatifida were added to the diet of juvenile barramundi, Lates calcarifer. The fucoidan extract was of 86% purity and included in two treatment diets at 5 g kg^?1 (0.5%) and 10 g kg^?1 (1%) respectively. Fish were fed the diet for 52 days. At the end of the feeding trial, barramundi fed with the 1% fucoidan inclusion diet exhibited enhanced growth with significantly increased lengths, weights and muscle fibre area (hypertrophy) when compared to the 0.5% fucoidan inclusion diet and the non‐fucoidan control diet. This research suggests that addition of fucoidan to the diet of fish may have growth promoting effects.     

Nutrient and amino acid profiles of egg and larvae of Asian seabass, Lates calcarifer (Bloch)

Fertilized eggs and developing larvae of hatchery reared Asian seabass, Lates calcarifer (Bloch), were analyzed to determine the changes occurring in their proximate and amino acid (AA) composition. The fertilized dry egg weighed 31 μg and contained 13.71 μg (44%) protein, 8.48 μg (27%) lipid and 0.657 J of gross energy. Dry weight decreased by 39% during hatching. The protein, lipid and carbohydrate nutrients decreased by 4.86, 4.15 and 0.09 μg, respectively from egg to 2-days post hatching (dph) larvae (pre-feeding). The protein content of the spawned eggs and larvae were hydrolysed to AA in the laboratory. The fertilized eggs had a total AA content of 42% of their dry weight. The egg contained 1.287 μg, 1.132 μg, 0.964 μg, 0.942 μg, 0.787 μg and 0.713 μg of leucine, lysine, arginine, valine, threonine and phenylalanine, respectively and these six indispensable amino acids (IAA) constituted approximately 78% of the total IAA. In the early feeding stages of L. calcarifer larvae, the ratio of IAA/DAA increased from 0.797 in the pre-feeding stage to 1.632 after 2 days of feeding. During larval growth of L. calcarifer, the percentage contribution of isoleucine and leucine to total IAA contents increased, while it decreased for lysine, phenyl alanine and arginine. L. calcarifer larvae were found to have proteins, which are rich in glutamic acid, leucine and lysine, and poor in threonine and histidine, suggesting high dietary leucine and lysine IAA requirement. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparison of the pathogenicity of Francisella orientalis in Nile tilapia (Oreochromis niloticus), Asian seabass (Lates calcarifer) and largemouth bass (Micropterus salmoides) through experimental intraperitoneal infection

Francisella orientalis is a highly virulent, emerging bacterium that causes mass mortalities in tilapia. This pathogen also affects numerous other warm-water fish species, including three-line grunt, hybrid striped bass and various ornamental fish. This study sheds light on two new species of fish that are susceptible to F. orientalis. Asian seabass and largemouth bass showed variable levels of susceptibility in a bacterial challenge experiment. After intraperitoneally injected with a dose of 10⁶ CFU/fish, a total of 64.28% and 21.42% mortalities were obtained in Asian seabass and largemouth bass, respectively. Meanwhile, Nile tilapia showed acute mortality of 100%. All fish showed typical lesions of francisellosis, including multifocal granulomas in the spleen and head kidney. Immunohistochemical analysis revealed strong positive signals inside the granulomas of all fish. The bacterial recovery in solid media from infected fish was highest in Nile tilapia (85.71%), followed by Asian seabass (35.71%) and largemouth bass (21.42%). PCR results tested 100% positive for Nile tilapia, and 78.57% and 21.42% for Asian seabass and largemouth bass, respectively. In conclusion, Asian seabass and largemouth bass are susceptible to this pathogen, which warrants new management strategies when employing predation polyculture systems of these species with tilapia.