Methods of microbial control in marine fish larval rearing: clay‐based turbidity and passive larval transfer

This study focused on methods to reduce bacterial loads in the larval culture tanks of California yellowtail (Seriola lalandi). We conducted two trials to evaluate methods to minimize bacterial loads in the larval rearing water. The first trial examined the use of bentonite clay as a turbidity agent to replace algae in a green water‐type environment. This trial consisted of three treatments: (1) clay with continuous feeding (CCO), (2) clay with batch feedings (CBA) and (3) algae paste with batch feedings (ALG). The results showed that both clay treatments had significantly fewer Vibrio colonies in the water column (CBA – 180 ± 78; CCO – 377 ± 120 CFU mL^?1) than the ALG treatment (5692 ± 2396 CFU mL^?1) after 14 days of culture. Survival was significantly higher in the CCO treatment (14.1 ± 2.6%) than either the CBA (2.3 ± 0.5%) or ALG treatments (2.8 ± 1.5%). The second trial attempted to limit bacterial loading in the larval culture tank by passively transferring the larvae into an adjacent, clean tank at 1, 5 and 9 days post hatch during the first 2 weeks of culture. The results from this trial showed that after 12 days of culture, water in the transfertank had fewer Vibrio colonies (1025 ± 541 CFU mL^?1) than the water in the control tanks (1962 ± 1415 CFU mL^?1). Also, survival was significantly higher among larvae that were transferred (43.9 ± 13.5%) than in the control tanks (23.1 ± 6.3%).     

Effect of dietary supplementation of periphyton on growth,immune response and metabolic enzyme activities in Penaeus monodon

A 60‐day indoor trial was conducted to study the effect of periphyton supplementation on metabolic and immune responses in tiger shrimp, Penaeus monodon. Periphyton developed over bamboo substrate in outdoor tanks (15 m²) was used as dietary supplement for P. monodon (2.02 ± 0.04 g) reared in 1000 L FRP tanks. Graded levels of periphyton were included in shrimp basal diets: 0% (P0), 3% (P3), 6% (P6), 9% (P9) and P0 diet with natural periphyton (NP) over bamboo substrate. At the end of the trial, P6 and NP showed significantly higher (P < 0.01) growth rate, 23.9% and 20%, respectively, compared with control, P0. Comparatively, lower level of metabolic enzymes, such as lactate dehydrogenase, malate dehydrogenase, aspartate aminotransferase and alanine aminotransferase, was recorded in treatments P3, P6 and NP compared with control, P0. The periphyton‐supplemented group, P3 had significantly higher (P < 0.05) superoxide dismutase (15.83 ± 0.96) and catalase activity (15.73 ± 0.69) compared to 6.88 ± 2.84 and 9.15 ± 0.67 unit mg^?1 protein min^?1_, respectively, in P0. Similarly, higher total haemocyte counts, 32.58 ± 1.30, 28.51 ± 3.12 and 27.26 ± 4.43 × 10⁶ cells mL^?1_, were recorded in P6, NP and P3, respectively, compared to P0, 23.57 ± 1.80 × 10⁶ cells mL^?1. After challenge with Vibrio harveyi, P3 recorded the highest relative percentage survival 67% followed by NP (58%) and P6 (42%) compared with control. However, treatment with highest periphyton inclusion (P9) did not differ significantly with P0 on growth and immunological parameters. This study indicates that periphyton supplementation at 3–6% level improves growth, immune response and metabolic activities in P. monodon.     

Stocking density effects on growth and production of the threatened silurid catfish,Mystus cavasius (Hamilton) fingerlings in nursery ponds

The fingerling‐rearing experiment of the threatened catfish, Mystus cavasius was carried out at different stocking densities in earthen nursery ponds. Twelve‐day‐old fry were stocked at 200 000 ha^?1 in treatment‐1 (T₁), 250 000 ha^?1 in treatment‐2 (T₂) and 300 000 ha^?1 in treatment‐3 (T₃) respectively. The mean length and weight of fry at stocking was 1.24 ± 0.25 cm and 0.11 ± 0.04 g respectively. Fry in all the experimental ponds were supplemented with SABINCO nursery feed for the first 14 days and starter‐I feed for days 15–56. The physico‐chemical parameters and plankton population of pond water were within the suitable level for fish culture. Growth in terms of final weight, final length, weight gain, length gain and specific growth rate and survival of fingerlings were significantly higher in T₁ than those in T₂ and T₃. Feed conversion rate was significantly lower in T₁ followed by T₂ and T₃ in that order. Significantly higher number of fingerlings was produced in T₃ than that in T₂ and T₁. Even then, consistently higher net benefits were obtained from T₁ than those from T₃ and T₂. Among the treatments evaluated, 200 000 fry ha^?1 was the best stocking density considering the highest growth, production and net benefits of fingerlings of M. cavasius in nursery ponds.     

Development and validation of a real‐time PCR assay for the detection of Aeromonas salmonicida

A real‐time PCR assay using a molecular beacon was developed and validated to detect the vapA (surface array protein) gene in the fish pathogen, Aeromonas salmonicida. The assay had 100% analytical specificity and analytical sensitivities of 5 ± 0 fg (DNA), 2.2 × 10⁴ ± 1 × 10⁴ CFU g^?1 (without enrichment) and 40 ± 10 CFU g^?1 (with enrichment) in kidney tissue. The assay was highly repeatable and proved to be robust following equivalency testing using a different real‐time PCR platform. Following analytical validation, diagnostic specificity was determined using New Zealand farmed Chinook salmon, Oncorhynchus tshawytscha (Walbaum), (n = 750) and pink shubunkin, Carassius auratus (L.) (n = 157). The real‐time PCR was run in parallel with culture and all fish tested were found to be negative by both methods for A. salmonicida, resulting in 100% diagnostic specificity (95% confidence interval). The molecular beacon real‐time PCR system is specific, sensitive and a reproducible method for the detection of A. salmonicida. It can be used for diagnostic testing, health certification and active surveillance programmes.     

Dietary biofloc supplementation in black tiger shrimp,Penaeus monodon: effects on immunity,antioxidant and metabolic enzyme activities

A 60‐day indoor experiment was conducted to study the effect of dietary supplementation of biofloc on metabolic enzyme activities and immune responses in Penaeus monodon juveniles. Biofloc developed in indoor fibreglass‐reinforced plastic (FRP) tanks (1000 L) was used as dietary supplement in P. monodon (2.90 ± 0.10 g) reared in 1000‐L FRP tanks. Graded level of dried biofloc was included in shrimp basal diets, 0% (control, B0), 4% (B4), 8% (B8) and 12% (B12). The level of metabolic enzymes like malate dehydrogenase (MDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) was not significantly different with control up to 8% dietary supplementation. A higher level of total haemocyte count (THC) was noticed in B8 (22.16 ± 2.17 × 10⁶ cells mL^?1) and B4 (21.11 ± 0.56 × 10⁶ cells mL^?1) compared with control, C (14.61 ± 2.74 × 10⁶ cells mL^?1). Biofloc‐supplemented groups recorded significantly higher (P < 0.05) serum SOD and catalase activity (P < 0.01) in comparison with control. The groups fed with 4% dietary biofloc supplement recorded highest relative percentage survival (RPS), 45% after challenge with Vibrio harveyi followed by 36% and 27% RPS in B8 and B12 groups. Based on these results, it can be concluded that supplementation of biofloc even at 4% level in the feed improves immune responses and metabolic activities in black tiger shrimp juveniles.     

Anaesthetic efficacy of MS‐222 and AQUI‐S® in advanced size fry of rohu,Labeo rohita, (Hamilton‐Buchanan)

Anaesthetic agents are very useful for reducing the stress caused by handling, sorting, transportation, artificial reproduction, tagging, administration of vaccines and surgical procedures in fish. The efficacy of two anaesthetics: MS‐222 and AQUI‐S^® were tested on rohu, Labeo rohita advanced size fry. The lowest effective doses that produced induction in 3 min or less and recovery times 5 min or less and meet the most criteria of good anaesthetic characteristics were 125 mg L^?1 of MS‐222, and 30 mg L^?1 of AQUI‐S^® in rohu, Labeo rohita advanced size fry. Induction times were significantly decreased with increased in the concentrations of any of the two tested anaesthetic agents. The lowest doses suitable for transportation of rohu advanced size fry observed were: 10–15 mg L^?1 of MS‐222 and 2.5 mg L^?1 AQUI‐S^®. Both anaesthetics showed promising to be used as anaesthetics for handling and transportation in rohu (Labeo rohita) advanced fry.     

Synbiotic effect of Bacillus mycoides and organic selenium on immunity and growth of marron,Cherax cainii (Austin, 2002)

The present feeding trial examined the effect of synbiotic use of Bacillus mycoides and organic selenium (OS) as Sel‐Plex on marron immunity, growth and survival. The marron were cultured in recirculated tanks and fed test diets consisting of a basal diet; basal diet supplemented with B. mycoides (10⁸ CFU g^?1 of feed); basal diet supplemented with OS (Sel‐Plex) (0.2 g kg ^{? 1} of feed) and basal diet supplemented with synbiotic (B. mycoides at 10⁸ CFU g^?1 and OS 0.2 g kg ^{? 1} feed) diet, in triplicate. The effect of the prebiotic OS (Sel‐Plex) on the growth rate of B. mycoides was also studied in vitro. The results suggested that synbiotic use of B. mycoides and OS significantly improved some immune parameters of marron, particularly the glutathione peroxidase, and to some extent total haemocyte counts. However, the synbiotic feed did not synergistically improve marron growth, in fact the use of B. mycoides‐supplemented diet alone demonstrated significantly higher growth in marron compared with the growth of marron fed on other test diets. Supplementation of the basal diet with host origin B. mycoides significantly increased the intestinal bacterial population (3.399 ± 825 CFU g ^{? 1} of gut) in marron compared with other diets. Organic selenium as Sel‐Plex in Trypticase Soya Broth also confirmed that OS did not increase the amount of growth of B. mycoides and resulted in a lower intestinal bacterial population in the synbiotic diet‐fed marron. In conclusion, synbiotic of OS and B. mycoides may improve a particular immune parameters of marron and to a lesser extent their growth.     

Combined effects of dietary fructooligosaccharide and Bacillus licheniformis on growth performance,body composition,intestinal enzymes activities and gut histology of triangular bream (Megalobrama terminalis)

This study was conducted to investigate the effects of fructooligosaccharide (FOS) and Bacillus licheniformis (B. licheniformis) on growth performance, body composition, intestinal enzymes activities and gut histology of Megalobrama terminalis. Nine experimental diets were formulated to contain three FOS levels (0, 3 and 6 g kg^?1) and three B. licheniformis levels (0, 1 and 5 × 10⁷ CFU g^?1) following a 3 × 3 factorial design. Accordingly, diets were named as 0/0, 0/3, 0/6, 1/0, 1/3, 1/6, 5/0, 5/3 and 5/6 (B. licheniformis/FOS). At the end of the 8‐week feeding trial, weight gain (WG) and specific growth rate (SGR) of fish fed 6 g kg^?1 FOS were both significantly (P < 0.01 and P < 0.05) higher than that of the other groups in terms of dietary FOS levels. Besides, WG and SGR of fish fed 1 × 10⁷ CFU g^?1 B. licheniformis were significantly (P < 0.05) higher than that of the control group in terms of dietary B. licheniformis levels. In addition, a significant interaction (P < 0.05) between dietary FOS and B. licheniformis was observed in finial weight, WG, SGR as well as the survival rate with the highest values all observed in fish fed diet 1/3. Hepatosomatic index, carcass lipid content, lipase activities as well as microvilli length increased significantly (P < 0.05) from 0 to 1 × 10⁷ CFU g^?1, but no significant difference (P > 0.05) was observed in terms of dietary FOS levels. In addition, a significant (P < 0.05) interaction of FOS and B. licheniformis was observed in both protease and Na⁺, K⁺‐ATPase activities with the highest value obtained in fish fed diet 1/3. The results indicated that the dietary applications of dietary FOS and B. licheniformis alone or in combination can significantly improve the growth performance, survival rate, intestinal enzymes activities as well as microvilli length of triangular bream. In addition, there is a significant interaction between dietary FOS and B. licheniformis. The best combination for this species is 3 g kg^?1 FOS with 1 × 10⁷ CFU g^?1 B. licheniformis.     

Resistance to vibriosis,production performance and tolerance to stress of the selected giant freshwater prawn (Macrobrachium rosenbergii)

The purpose of this study was to evaluate the resistance to vibriosis, growth, survival and tolerance to stress of the selected prawn, second generation, compared to a non‐selected control. The first generation of selected giant freshwater prawn, which has 10.4% higher of resistance, was used to attain disease resistant freshwater prawn (Macrobrachium rosenbergii) generation through challenge test‐based selection. Resistance test was conducted by infecting the prawn (mean body weight of 10.29 ± 1.40 g) with pathogenic Vibrio harveyi (5 × 10⁵ cfu prawn^?1). The growth and survival of the prawn were evaluated by rearing the two populations of prawn in both nursery and grow‐out phases. Stress tolerance test was done by evaluating the viability of postlarvae exposed to environmental stressors, i.e. temperature, salinity, NH₃ and formaldehide. Post‐challenge survival of the selected prawn (55.0 ± 5.0%) was about 46% higher than that of the control (37.5 ± 7.5%). The survival of the selected prawn in nursery culture (77.16 ± 0.841%) was significantly higher (p < .05) than that of the control (51.31 ± 2.938%), while the survival in grow‐out culture was similar (p > .05). The growth of selected prawn (4.99 ± 0.03% day^?1) was significantly higher than that of the control (4.81 ± 0.05% day^?1). There was no difference between treatments on the tolerance level against the tested environmental stressor. Overall data suggested that the selected prawn showed better performance in growth and resistance against vibriosis.     

Effects of supplementary fennel (Foeniculum vulgare) essential oil in diet on growth and reproductive performance of the ornamental fish,Convict cichlid (Cichlasoma nigrofasciatum)

This study evaluated the effects of diet containing 0 (C: control), 75 (D₁), 100 (D₂), 125 (D₃) and 150 (D₄) mg kg^?1 F. vulgare essential oil on growth and reproductive performance of C. nigrofasciatum. A total of 225 Convict cichlids female with mean weight (1.65 ± 0.02 g) were distributed into 15 glass aquaria (15 fish tank^?1) in triplicate treatments. During 40 days of experimental period, fish fed at 3% of their body weight daily. At the end of experiment, growth indices, gonadosomatic index and reproductive indices were determined. The best feed conversion ratio was obtained in D₄ (1.19 ± 0.03, P < 0.05). The addition of F. vulgare essential oil did not have any statistical effects on other growth indices (specific growth rate, weight gain and condition factor). Total survival rates in all treatments and control were uniformly high, ranging from 93% to 97% (P > 0.05). F. vulgare essential oil has no negative effect on survival rate of C. nigrofasciatum. There were no significant differences among protein content of fish body (P > 0.05). The lowest value of fat content (26.5 ± 0.5%) was observed in D_2, and it had significant difference with control and other groups (P < 0.05). The ash percentage in treatments fed with F. vulgare essential oil were higher than control fish (P < 0.05). The highest moisture content was detected in D₄ (72.68 ± 0.19%) and D₂ (71.23 ± 2%) groups. Among the used dosage, D₄ was the most effective dosage that could significantly increase GSI (11.06 ± 1.55%), fecundity (340 ± 21) and hatching ratio (92.33 ± 1.63%). There were no significant differences in diameter of eggs among various treatments (P > 0.05).     

Immune responses and gut morphology of Senegalese sole (Solea senegalensis,Kaup 1858) fed monospecies and multispecies probiotics

The current study aimed to determine the effects of dietary probiotic supplementation on growth, gut morphology and non‐specific immune parameters in Senegalese sole (Solea senegalensis) juveniles during a 1‐month trial. Fish were fed for 1‐month two diets with 1.0 or 4.6 × 10⁶ CFU kg^?1) of probiotic A (Bacillus sp., Pediococcus sp., Enterococcus sp. and Lactobacillus sp.) and two diets with 3.5 or 8.6 × 10⁵ CFU kg^?1 of probiotic B (Pediococcus acidilactici) and tested against an unsupplemented diet (control). Growth performance, as well as respiratory burst activity, nitric oxide (NO), alternative complement pathway (ACH50), lysozyme and peroxidase activities, was not affected by the dietary treatments. Probiotic supplementation tended to increased growth homogeneity between tanks having diet A₁ the best possible alternative to decrease costs associated to size grading. Villous length and number of goblet cells of the anterior intestine did not vary among treatments. Muscle duodenal layer was significantly thicker in fish fed probiotic A compared to probiotic B, when included at the lowest level (A₂ versus B₂). The current study indicate that the use of the multispecies probiotic at 1.0 × 10⁶ CFU kg^?1 might enhance protection against pathogen outbreak and increase nutrient absorption, whereas at the highest concentration could reduced size dispersion among tanks.     

Histological alterations in gills of shrimp Litopenaeus vannamei in low‐salinity waters under different stocking densities: Potential relationship with nitrogen compounds

Two experimental modules with different stocking densities (M1 = 70 and M2 = 120 shrimp /m²) were examined weekly over a culture cycle in tanks with low‐salinity water (1.9 g/L) and zero water exchange. Results showed survival rates of 87.7 and 11.9% in M1 and M2, respectively. Water temperature, pH, dissolved oxygen, electrical conductivity and chlorophyll a were not significantly (p > .05) different between modules. In contrast, the concentrations of nitrogen compounds were significantly (p < .05) different between modules, except nitrite‐N (M2 were 2.31 ± 1.38 mg/L N‐TAN, 0.18 ± 0.49 mg/L N‐NO₂^? and 6.83 ± 6.52 mg/L N‐NO₃^?; in M1: 0.97 ± 0.73 mg/L N‐TAN, 0.05 ± 0.21 mg/L N‐NO₂^? and 0.63 ± 0.70 mg/L N‐NO₃^?). When waters of both modules reached higher levels of ammonia and nitrite, histological alterations were observed in gills. The histological alterations index (HAI) was higher in M2 (5‐112) than in M1 (2‐22).     

Studies on bacterial pathogens isolated from diseased torafugu (Takifugu rubripes) cultured in marine industrial recirculation aquaculture system in Shandong Province,China

In spring of 2011, an epidemic outbreak of torafugu with high mortality occurred in an aquafarm with marine industrial recirculation aquaculture system (MIRAS) in Yantai, Shandong Province, China. The diseased fish showed anorexia, haemorrhaging and festering fin and skin and swelling internal organs. Forty‐five dominant bacterial isolates were obtained from the diseased fish, and were found to belong to 12 species according to 16S rRNA gene sequences. One strain from each species was selected to test the pathogenicity, and five strains were showed to be virulent to zebrafish. Whereas Enterovibrio nigricans Fr42 was highly virulent with the LD₅₀ of 7.8 × 10⁴ CFU g^?1, Photobacterium swingsii Fr23, Vibrio owensii Fr40, V. harveyi Fr51 and V. rotiferianus Fr71 were moderately virulent with the LD₅₀ of 1.7 × 10⁶ to 8.4 × 10⁶ CFU g^?1. Both the bacteria and their extracellular products of the five strains were found to show phospholipase, caseinase, gelatinase, amylase and/or lipase activities. The production of N‐acyl homoserine lactones (AHLs) of the five strains was detected by three different AHLs biosensors, and three of them were found to produce AHLs by at least one kind of biosensor. This is the first study describing various opportunistic bacterial pathogens of fish cultured in MIRAS in China.     

Fertilization by refrigerator stored sperm of the Indian major carp, Labeo calbasu (Hamilton, 1822)

This study reports on the spermatological properties, and on the development of a protocol for refrigerator storage (4°C) of Labeo calbasu (Hamilton, 1822) sperm for artificial breeding. Volume, motility, concentration and pH of the freshly collected sperm were 2.21 ± 0.53 (μL g^?1 of fish weight) (mean ± SD), 95 ± 1 (%), 1.93 ± 0.44 × 10⁹ (cells mL^?1) and 7.56 ± 0.17 respectively. Sperm activation was evaluated at different osmolalities of NaCl solution, and motility ceased completely when osmolality of the extender was ≥287 mOsmol kg^?1. Sperm retained motility for 24, 72 and 108 h, after refrigerator storage when sperm were undiluted, suspended in Alsever's solution and suspended in Alsever's solution containing 5% methanol respectively. Fertilization rate of fresh eggs with sperm stored at 4°C in Alsever's solution and Alsever's solution containing 5% methanol was 77% and 60% with a hatching rate of 60% and 43% respectively. The fertilization and hatching success of the stored sperm suggests potential to use refrigeration for transporting genetic material to hatcheries for artificial breeding of L. calbasu in Bangladesh.     

Evaluation of live duckweed,Wolffia globosa as an allochthonous feed for Labeo rohita fry during nursery rearing

A 40 days feeding trial with Labeo rohita (rohu) fry was conducted in six outdoor cemented tanks (5m × 4m × 1 m; 6–8 cm soil base) to establish the relative impact of on‐farm grown live Wolffia globosa (a duckweed with crude protein of 28.57%) and a formulated iso‐nitrogenous feed (crude protein 28.37%) on growth performance, survival, health and overall quality of fingerlings. The experimental tanks were dried, limed at 250 kg/ha, filled with ground water and subsequently applied with cow dung (soaked overnight) and mustard oil cake at 1,000 kg/ha and 50 kg/ha respectively. The fry were stocked at 0.3 millions/ha in six prepared tanks, and fed live Wolffia (T1) and formulated feed (T2). Results of the study indicate better performance (p < 0.05) of fry in terms of final mean weight (7.83 g), length (8.82 cm), daily growth index (6.51 g %/day) and survival (89.5%) than those fed on formulated feed. Furthermore, significantly improved (p < 0.05) feed utilization parameters viz. apparent feed conversion ratio (AFCR), apparent protein conversion efficiency (APCE), apparent protein efficiency ratio (APER); haematological parameters viz. total erythrocyte count (TEC), total leucocyte count (TLC), packed cell volume (PCV), plasma glucose and protein of fry were also observed in T1 (live Wolffia). Overall findings from this study hints towards the possible utilization of on‐farm grown live Wolffia as a suitable nutrient source for fry rearing of L. rohita.     

Nano zinc vis‐à‐vis inorganic Zinc as feed additives: Effects on growth,activity of hepatic enzymes and non‐specific immunity in rohu,Labeo rohita (Hamilton) fingerlings

The effects of dietary administration of inorganic zinc (zinc sulphate, ZnSO₄) and nano zinc (zinc oxide nanoparticles, ZnO‐NP) were evaluated in rohu, Labeo rohita fingerlings. Fish were fed with a basal diet (Control) supplemented with ZnSO₄ (T1, T2 and T3) and ZnO‐NP (T4, T5 and T6) at 10, 20 and 30 mg/kg, respectively, for a duration of 45 days. The results revealed that fish fed diet containing 20 mg ZnO‐NP per kg (T5) had the highest weight gain and specific growth rate (SGR, % per day), which was significantly different (p < .05) from the other experimental diets. Significantly (p < .05), higher activities of the digestive and metabolic enzymes were recorded in the fish fed ZnO‐NP containing diets as compared to the diets containing inorganic Zn or control diet. The maximum serum glucose and protein levels were noted in fish reared on diet T5. Both SGOT and SGPT activities were significantly increased in fish fed Zn‐supplemented diets (T1 to T6), as compared to the control group. Similarly, innate immune parameters were improved with feeding Zn incorporated diets. The highest phagocytic (40.74 ± 0.65%) and respiratory burst (0.33 ± 0.001, OD 630nm) activities were recorded in the fish fed diet containing ZnO‐NPs at 20 mg/kg (T5). The maximum superoxide production and serum peroxidase activity were detected in the fish fed T5 and T6 diets. Overall, results indicated that short‐duration feeding (≤45 days) of dietary ZnO‐NP (20 mg/kg) improved growth, enzyme activity, serum biochemical parameters and immune function in rohu fingerlings.     

Effects of different carotenoid supplementation sources with or without sodium taurocholate on growth,feed utilization,carotenoid content and antioxidant status in fry of the European seabass,Dicentrarchus labrax

A total of 210 European seabass fry with an average body weight of 0.51 ± 0.03 g were divided into seven experimental treatments (three replicates per treatment). The seven experimental diets (45.64% CP) were formulated to contain synthetic astaxanthin (SA; 0.1 g kg^?1), marigold flower meal (MFM; 0.2 g kg^?1) or crab waste meal (CM; 10 g kg^?1), with or without sodium taurocholate (ST; 0.4 g kg^?1); the control diet contained no supplementation. The experiment was conducted for 8 weeks. The results show that seabass fry fed the diets containing different sources of carotenoids, especially with ST, demonstrated higher growth efficiency, survival (%), feed efficiency, protein and ether extract contents of fish body. The best results were found with Diet_MFM+ST. Whole‐body carotenoid levels increased significantly with all supplementation treatments; the highest level was observed with SA, followed by fish fed Diet_MFM and Diet_CM, especially with ST co‐supplementation. Moreover, thiobarbituric acid‐reactive substances were significantly reduced in fish fed the diets containing different sources of carotenoids compared to the control diet. The total antioxidant status significantly increased in fish fed Diet_SA and Diet_CM alone or with ST compared to the control group. In conclusion, these results reveal that Diet_MFM+ST is the most suitable treatment for improving growth, feed efficiency, and the protein and ether extract contents of seabass fry compared to other sources of carotenoids. The best oxidant/antioxidant balance and carotenoid content were achieved using SA, CM and MFM with ST supplementation.     

Preliminary study on broodstock rearing,induced breeding and grow‐out culture of the sea cucumber Holothuria scabra in Sri Lanka

Experiments were conducted to identify suitable methods for broodstock rearing, induced breeding and grow‐out culture of Holothuria scabra in Sri Lanka. Two hundred and seventy‐two brooders (500–600 g) collected from off Mannar were individually packed in oxygen‐filled polythene bags with and without sea water and transported to a sea cucumber hatchery at Kalpitiya. Lagoon pens, sand‐filled fibreglass tanks and bare tanks were used in triplicates to maintain brooders. Spawning was initiated using air dry, water jet and thermal‐stimulation methods. Hatchery produced juveniles with an average weight of 11 ± 5 g were reared (2 individuals m^?2) in lagoon pens, mud ponds and fibreglass tanks in triplicates. The significantly high evisceration rate was observed when brooders were transported without sea water (t‐test, P < 0.05). Brooders maintained in bare tanks showed a significant weight reduction than the brooders in sand‐filled tanks and lagoon pens (anova , P < 0.05, d.f. = 2). Thermal stimulation (ambient temperature ± 3–5°C) was found to be the most successful method of spawning initiation of H. scabra. The mean (±SD) percentage males and females participated for spawning per trial was 9.2 (±10) and 4.6 (±5.6) respectively. On an average, 1.16 millions of eggs (±1.03 SD, n = 5) were obtained per spawning trial. H. scabra juveniles reared in tanks showed significantly lower growth rate than the juveniles in pens and ponds (anova , P < 0.05). Lagoon pens and sand‐filled tanks are suitable to maintain brooders and lagoon pens can be successfully used for mass rearing of juveniles.     

Short‐term exposure to higher temperature triggers the metabolic enzyme activities and growth of fish Labeo rohita fed with high‐protein diet

Six hundred Labeo rohita (average weight, 6.78 g) were distributed in 24 tanks (25 fish per tank), which were allotted to eight experimental groups in triplicates. Half of the experimental groups were maintained at ambient water temperature of 26 °C, whereas the other half were exposed to 32 °C for 1 week then later maintained at 26 °C for 4 weeks. Fish reared under different temperature regimes were fed one of the four diets containing 200 g kg^?1, 300 g kg^?1, 400 g kg^?1 or 450 g kg^?1 protein. Growth, feed efficiency and protein retention were higher at 32 °C and continued for the next 2 weeks after decreasing the temperature. The dietary protein level and water temperature interactions were more effective in modulating the response of hepatic glucokinase than that of hexokinase and pyruvate kinase. In contrast, hepatic gluconeogenesis was not affected by rearing temperature. Hepatic aspartate aminotransferase and alanine aminotransferase activity was found higher (P < 0.05) at low temperature and at high protein level. It is suggested that the metabolic activities of fish are triggered by short‐term exposure to higher temperature and the increased metabolic activity extended for a duration of 3 weeks during which 400 g kg^?1 dietary protein level was found to support growth of fish.     

Microcystis aeruginosa (Kütz) incorporated diets increase immunity and survival of Indian major carp Labeo rohita (Ham.) against Aeromonas hydrophila infection

The aim of this study was to evaluate dietary dosages of Microcystis on the immune response and disease resistance against infections due to the opportunistic pathogen Aeromonas hydrophila in Labeo rohita fingerlings. Labeo rohita fingerlings were fed diet containing 0 (Control), 0.5, 1.0 and 5 g Microcystis powder kg^?1 dry diet for 90 days. Three replicate groups of fish averaging 20 ± 2 g were fed for 3 months daily. At 30 days interval, samples were assayed for different biochemical [serum total protein, albumin, globulin, albumin:globulin (A:G) ratio] and immunological (superoxide anion production, lysozyme and serum bactericidal activity) parameters. The results demonstrate that fish fed Microcystis showed increased levels of lysozyme, serum bactericidal activity, serum protein and albumin (P ≤ 0.05) as compared with the control group. After 90 days, fish were challenged with A. hydrophila and mortality (%) was recorded up to day 10 postchallenge. The group fed 1.0 g Microcystis kg^?1 dry diet showed the highest percentage survival (72%). These results indicate that Microcystis aeruginosa stimulates the immunity and makes L. rohita more resistant to infection by A. hydrophila when fed in dried form in feed.