Probiotic activity of Bacillus subtilis in juvenile freshwater prawn, Macrobrachium rosenbergii (de Man) at different methods of administration to the feed

This experiment was carried out to investigate the potential probiotic properties of Bacillus subtilis and suitable methods of administration to the commercial feed in juvenile Macrobrachium rosenbergii (de Man) from 25 August to 25 November 2008. Putative Bacillus subtilis bacterium isolated from juvenile M. rosenbergii intestine was added to commercial prawn feed as a probiotic. Five types of diets were prepared by mixing B. subtilis at level of 108 cells g^?1 with commercial feed using different methods consisting mixing, soaking, spraying and bathing. After 60?days, the prawns fed diets at soaking method treated group, showed a higher mean weight gain (2.09) or 328.84% increase in growth over control. There were significant differences (P?<?0.05) in weight gain, feed intake and FCR among soaking method and other treated and control groups. There was significant difference (P?<?0.05) in survival among treated and control groups, but no significant difference (P?>?0.05) in water quality and biochemical composition among treated and control groups. Clearly, B. subtilis-treated diets appeared to enhance growth and survival of juveniles M. rosenbergii. It was concluded that the tested strain may be a promising probiotic for M. rosenbergii under soaking method of administration to the prawn commercial feed.     

Effect of treatment with probiotics as water additives on tilapia (Oreochromis niloticus) growth performance and immune response

A feeding trial was conducted for 40 days to delineate the effect of treatment with probiotics as water additives on tilapia (Oreochromis niloticus) growth performance and immune response. About 360 juveniles were randomly distributed into four treatment groups, each with three replicates. Different probiotics (T-1, Bacillus subtilis B10; T-2, Bacillus coagulans B16; T-3, Rhodopseudomonas palustris G06) were added to the water of tanks at final concentration of 1 × 10⁷ cfu ml^?1 every 2 days, with no probiotic added to control tanks. At the end of the feeding trial, fish treated with B. coagulans B16 (T-2) and R. palustris G06 (T-3) had significantly (P < 0.05) higher final weight, daily weight gain, and specific growth rate compared with those treated with B. subtilis B10 (T-1) and those without probiotics (control). The highest (P < 0.05) content of total serum protein was found in T-2 compared with that in T-1, T-3, and the control. However, albumin concentration and albumin/globulin ratio were not affected by the probiotics treatments. Compared with the control, probiotic supplementation remarkably improved activities of superoxide dismutase and catalase (P < 0.05). T-2 fish exhibited higher average myeloperoxidase activity than the control, T-1, and T-3 groups. Regarding serum lysozyme content in tilapia, assays showed no difference (P > 0.05) among the treatment groups. Furthermore, probiotics treatments remarkably increased respiratory burst activity compared with control, with T-2 showing higher values than T-1 and T-3. This indicated that treatment with probiotics, B. coagulans B16 and R. palustris G06, as water additives could be used to enhance immune and health status, thereby improving growth performance of O. niloticus.     

Effect of dietary supplementation of Bacillus licheniformis on gut microbiota, growth and immune response in giant freshwater prawn, Macrobrachium rosenbergii (de Man, 1879)

The present study was conducted to evaluate the effect of dietary supplementation of probiotic bacterium Bacillus licheniformis on the gut microbiota, growth and immune response in Macrobrachium rosenbergii juveniles (4.0 ± 0.02 g). Five isocaloric experimental diets supplemented with different concentrations of B. licheniformis, viz. Control (without probiotic supplementation), T₁ (1.0 × 10⁶ cfu g^?1feed), T₂ (1.0 × 10⁷ cfu g^?1feed), T₃ (1.0 × 10⁸ cfu g^?1 feed) and T₄ (1.0 × 10⁹ cfu g^?1 feed) were used for the experiment. After 60 days of feeding trial, the growth performance increases significantly in all the experimental groups compared with control, and highest weight gain, specific growth rate and lowest feed conversion ratio were recorded from T₄ (P > 0.05). However, juvenile survival was not found to be significantly affected by probiotic supplementation (P > 0.05). The total bacterial count and Bacillus spp. count in the gut were significantly increased with concurrent decrease in pathogenic Aeromonas spp. and Pseudomonas spp. in all the experimental groups compared with control (P < 0.05). Compared with control, immune parameters were increased significantly in all the experimental groups (P < 0.05). Highest haemocyte count, phenol oxidase, superoxide dismutase and antibacterial activity in haemolymph were recorded from T₄ (P < 0.05). When prawns were challenged with Vibrio alginolyticus, the cumulative mortality was significantly lower in T₄ (P < 0.05). These results indicate that dietary supplementation of B. licheniformis can manipulate the gut microbiota and increase the growth and immune response of M. rosenbergii. In the present study, concentration response on the probiotic effect of B. licheniformis was observed which showed higher effects at a concentration of 1.0 × 10⁹ cfu g^?1 feed in the laboratory condition.     

Effect of dietary mannan oligosaccharide (MOS) on growth performance, survival, body composition, and some hematological parameters in giant sturgeon juvenile (Huso huso Linnaeus, 1754)

The effect of dietary mannan oligosaccharide (MOS; activeMOS^®) on growth, survival, and body composition in giant sturgeon juvenile (Huso huso) with initially average weight 46.89 ± 2.57 was investigated for a period of 46 days. Basal diet were supplemented with 0 (control), 2, and 4 g kg^?1 MOS in a totally randomized design trial in triplicate groups. The results showed no significant differences in growth and feeding parameters between control and treatment groups (MOS supplementation diets) (P > 0.05). There was a statistically significant decrease (P < 0.05) in feed per fish level in only group treated with 4 g kg^?1 MOS. The highest and the lowest growth performances were observed in 2 and 4 g kg^?1 MOS, respectively. There were no significant differences in survival rate among all treatment groups (P > 0.05). In group treated with 2 g kg^?1 MOS was a significant difference in lipid carcass (P < 0.05), whereas protein, ash, and moisture remained unaffected (P > 0.05). However, no significant difference was observed in intestinal lactic acid bacteria between all treatment groups (P > 0.05). There were no significant differences in hematological parameters between control and MOS treatment groups (P > 0.05). These results suggested that the prebiotic mannan oligosaccharide did not influence the growth performance in giant sturgeon juvenile, and it is not appropriate for supplementation in the diet of cultured juvenile giant sturgeon.     

Growth enhancement and protective potential of feed-based outer membrane proteins against vibriosis in <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis>

The use of antibiotics to curtail vibriosis, which is a major infectious disease, plaguing shrimp and prawn is rather becoming less effective and the need for a better alternative is expedient. The outer membrane proteins (OMPs) of V. alginolyticus were extracted, mixed with powdered commercial feed and fed to the prawns to evaluate its effect on growth performance and protective potential. Sixty prawns were divided into groups A, B and C of 10 prawns each, with two replicates in six (150 L) glass aquaria. Groups A, B and C were fed with OMPs mixed diet, with OMPs-Freund’s incomplete adjuvant mixed diet and OMPs or adjuvant free diet (control diet) respectively. All the prawns were weighed weekly, and haemolymph was collected to determine the total haemocyte count (THC) and phenoloxidase (PO) activity. At the end of the feeding trial, prawns were intramuscularly challenged with 50 μL of 10⁷ CFU V. alginolyticus. The treated groups were significantly higher in growth performance and THC than the control group, but no significant difference between the groups in terms of PO activity and mortality rate. The study, however, submitted that oral administration of OMPs with or without adjuvant is a good growth promoter and has the potential for protection against vibriosis in giant freshwater prawn (Macrobrachium rosenbergii).     

Improvement of intestinal barrier,immunity, and meat quality in common carp infected by Aeromonas hydrophila using probiotics

The fish pathogen Aeromonas hydrophila causes gastrointestinal tract infections and hemorrhagic septicemia and represents a widespread risk in aquaculture. This study aimed to determine whether compound probiotics could improve the intestinal barrier, immunity, and meat quality of common carp infected by A. hydrophila by feeding them compound probiotics. Carp were assigned to one of four groups: (1) control (no infection, no probiotics); (2) control?+?probiotic; (3) A. hydrophila infected; and (4) the A. hydrophila?+?probiotic group. At the beginning of the experiment, the carp were injected with either saline (0.86%) or A. hydrophila (4.87?×?10⁷ CFU/mL). After 2 weeks of the feeding regime, results suggested that in A. hydrophila infected carp, dietary probiotics regulated the intestinal microflora as evidenced by a reduced abundance of Proteobacteria and increased amounts of Firmicutes and Fusobacteria in the intestine. In addition, dietary probiotics ameliorated both villus swelling and the decrease in villus height induced by A. hydrophila (P?<?0.05). Moreover, probiotics prevented the A. hydrophila-induced decline of Occludin, Claudin-1, and ZO-1 levels in the intestine (P?<?0.05). The addition of probiotics into the feed also increased acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM) activity in the serum (P?<?0.05), in comparison to the A. hydrophila group. Importantly, when compared to A. hydrophila-infected carp, compound probiotics alleviated the decrease in muscle nutrient quality (P?<?0.05). In summary, we show that dietary compound probiotics can prevent the impact of A. hydrophila infection on the intestinal function and disease resistance, while ensuring carp muscle quality.

     

Dietary supplementation of yeast (<Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>) improves growth,stress tolerance,and disease resistance in juvenile Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

The yeast Saccharomyces cerevisiae is one of the commonest probiotics incorporated in aquafeeds. An 84-day feeding trial was conducted to evaluate the effects of varying dietary inclusions of S. cerevisiae, 0% (control), 3% (YF3), 5% (YF5), and 7% (YF7), on growth, stress tolerance, and disease resistance in juvenile (body mass ~?21 g) Nile tilapia (Oreochromis niloticus). Fish were randomly distributed in groups of 20 into 12 1-m³ hapas and fed isoenergetic (~?17 kJ g^?1 gross energy) and isonitrogenous (~?300 g kg^?1 crude protein) diets at 3% of their bulk weight daily. Specific growth rates were significantly higher for the yeast-fed fish (0.77–0.78% day^?1) than for the control fish (0.60% day^?1) and resulted in significantly higher mean final weights for the yeast-treated groups. Protein and lipid retention efficiencies were also significantly higher in the yeast-fed fish than in the control group. In subsequent stress challenge trials, the yeast-fed fish had greater tolerance to acute heat as well as hypoxia exposure than the control fish. Survival rates of the yeast-treated groups following sudden exposure to elevated water temperature (40 °C) ranged from 82.5 to 100% compared to 15% for the control. Mean survival rates following a 24-h hypoxia exposure were also generally significantly higher for the yeast-fed fish. The probiotic groups recorded a relative percent survival (RPS) of 75% after a 14-day Aeromonas hydrophila infection challenge. The results of this study indicate that S. cerevisiae as an additive in Nile tilapia diets has beneficial impacts on growth, stress tolerance, and disease resistance.     

Combined diet of yeast,fermented soybean meal,and microparticulate as larval feed in extensive rearing systems for seed production of the oriental river prawn <Emphasis Type="Italic">Macrobrachium nipponense</Emphasis>

The present study aims to evaluate the effects of several types of larval feed on seed production of the oriental river prawn Macrobrachium nipponense and water quality in extensive rearing systems. Healthy M. nipponense were distributed into fifteen 15-m² pond outdoors, and each pond was stocked with 90 prawns including 41 egg-bearing (berried) female prawns, 37 not bearing female prawns, and 12 male prawns for larval production. The experimental design was completely randomized, with four diet groups (diet A: soybean homogenate, diet B: yeast, diet C: yeast?+?fermented soybean meal, and diet D: yeast?+?fermented soybean meal?+?microparticulate diet) and one no feeding control and three replications. There are equal levels of the total protein content in diets A–D. Higher larvae and postlarvae number was obtained for diet C and diet D. Furthermore, the increase of M. nipponense larvae and postlarvae in diets B–D can result in the decrease of zooplankton and increase of phytoplankton in the pond during some periods, simultaneously improving the water quality. The Spearman correlation coefficient (r_M,N) between the amounts of larvae and postlarvae of M. nipponense per pond and zooplankton per pond during the entire experiment period was ??0.56. The highest juvenile M. nipponense production was obtained in the diet D group (3140.00?±?461.63), which was slightly higher than that in the diet C group (2940.33?±?382.60). The juvenile M. nipponense production in diet B (2303.00?±?449.23) was also significantly higher compared to diet A (1106.33?±?176.64) and the control group (1039.67?±?212.85). There were no significant differences in the amounts of juvenile M. nipponense between the control and diet A groups, which indicated that the soybean homogenate could not increase the amount of M. nipponense juveniles. Our results clearly showed that diet D significantly raised per unit yield of prawn seed comparing with no feeding and other diet group in extensive rearing systems and that the cost of producing prawn seed with diet D is low, which provide an important foundation for the expansion of oriental river prawn culture.     

Dietary intake of probiotics and maslinic acid in juvenile dentex (Dentex dentex L.): effects on growth performance, survival and liver proteolytic activities

Two feeding trials were carried out to evaluate the efficiency of probiotics and maslinic acid, on growth and survival of juvenile dentex; liver proteolytic activities were also investigated in the second trial. For experiment 1, triplicate groups were fed six diets with two probiotics (Bacillus toyoi, T, and B. cereus, E) at increasing levels (0.5, 1 and 2 g kg^?1 diet) and a control diet. Growth and feed conversion were not significantly influenced by the probiotics. The diet T1 produced the lower mortality, whereas diet E1 rendered the higher mortality. It was concluded that no significant effects on growth and survival were found following the addition of two kinds of probiotics to dentex diets. However, the diet E0.5 showed a tendency to ameliorate the growth and feed utilization of the diet. In a second trial, triplicate groups were fed four diets with increasing levels of maslinic acid (0, 20, 40 and 80 mg kg^?1 diet). Growth of fish given diets with the highest level of maslinic acid (D80) was slightly but not significantly lower than those from the other groups. Furthemore, mortality of fish fed diet D40 was the lowest. Changes in liver proteasome and endoprotease activities measured on sodium dodecyl sulphate (SDS)/gelatin gels were also detected in a dose‐dependent manner. It was concluded that a dietary maslinic acid at a level of 80 mg kg^?1 diet seems to be too high for juvenile dentex to maintain a maximal growth and survival rate.     

Effects of cholesterol on growth,feed utilization,body composition and immune parameters in juvenile oriental river prawn,Macrobrachium nipponense (De Haan)

An 8‐week feeding trial was conducted to investigate the effects of dietary cholesterol levels on growth, feed utilization, body composition and immune parameters in juvenile oriental river prawn, Macrobrachium nipponense. Six isolipid (80 g kg^?1 crude lipid) and isoproteic (400 g kg^?1 crude protein) diets, supplemented with 0, 3.0, 6.0, 9.0, 12.0 and 15.0 g kg^?1 cholesterol, were evaluated. Growth performance and feed utilization of M. nipponense were improved as dietary cholesterol levels increased. Weight gain and specific growth rate were highest, and feed conversation ratio was lowest, when prawns were fed a diet supplemented with 9.0 g kg^?1 cholesterol. However, final body weights and survival rates of juvenile M. nipponense were not affected significantly by dietary cholesterol. Body composition of prawns, including moisture, crude protein and crude lipid, was not significantly affected by changes in dietary cholesterol. The immune parameters measured in hepatopancreas, including total antioxidant capacity, and glutathione, catalase, alkaline phosphatase and acid phosphatase activities, were at optimum levels in prawns fed with 9.0 g kg^?1 dietary cholesterol. In summary, the best growth performance, lowest feed conversation ratio, and the most enhanced antioxidant capacity and immunity parameters were attained in juvenile M. nipponense when fed a diet supplemented with 9.0 g kg^?1 cholesterol.     

Effect of Coriolus versicolor polysaccharides on the hematological and biochemical parameters and protection against Aeromonas hydrophila in allogynogenetic crucian carp (Carassius auratus gibelio)

The effect of dietary intake of Coriolus versicolor Polysaccharides (CVP) on the hematological and biochemical indices of Allogynogenetic crucian carp (Carassius auratus gibelio) was investigated. Fish were fed CVP supplemented diets (0, 0.25, 0.5, 1.0, 2.0 or 4.0 g CVP kg^?1) for 56 days. The RBC, WBC counts, hemoglobin content, ESR in blood and TP, ALT, AST, ALP, GLU, CHO, TG, and BUN in serum were measured on day 0, 14, 28, 42, and 56. After feeding of 56 days, fish were infected with Aeromonas hydrophila and mortalities were recorded. The results indicated that feeding crucian carp with suitable dose of CVP enhanced the RBC, WBC counts, hemoglobin and TP content, ALP activity, and decreased the ESR, ALT, AST, GLU, CHO, TG and BUN. There was no effect in fish at low dose (0.25 g kg^?1). Unexpectedly, the higher CVP dose used here (2.0 and 4.0 g kg^?1) has a negative effect in fish. The results of challenge experiment indicated that a moderate level of CVP in the diet (1.0 g kg^?1) was the most effective to enhance the survival of fish after infected with A. hydrophila. In summary, the use of CVP, as dietary supplements, can improve the innate defense of crucian carp providing resistance to pathogens.     

Evaluation of Bacillus subtilis as a probiotic to Indian major carp Labeo rohita (Ham.)

Bacillus subtilis, a Gram‐positive, aerobic, endospore‐forming bacterium, was evaluated for its probiotic potential in Indian major carp, Labeo rohita. Labeo rohita (15±2 g) were fed a feed containing B. subtilis in three concentrations for 2 weeks, e.g., 0.5 (T₂), 1.0 (T₃) and 1.5 (T₄) × 10⁷ CFU g^?1 feed. The control group (T₁) was fed feed without B. subtilis for the same period. Haematological and serum parameters were monitored at weekly intervals. The response variables were total erythrocyte count, total leucocyte count (TLC), haemoglobin, total protein, albumin, globulin, albumin–globulin ratio, alkaline phosphatase activity, alanine aminotransferase activity and aspartate aminotransferase activity. Fish were challenged intraperitoneally with a virulent strain of Aeromonas hydrophila after 2 weeks of feeding to the treatment groups and positive control group, while the negative control group was challenged with phosphate‐buffered saline only. Clinical signs and symptoms, and mortality/survival percentage were noted in each group. The haematological and serum parameters were monitored each week and during post challenge on the third and tenth day. The B. subtilis‐treated fish (T₄, 1.5 × 10⁷ CFU g^?1 feed) showed maximum per cent survival (87.50%), weight gain (35.5%), TLCs (3.23 × 10⁴ cells mm^?3), haemoglobin content (7.4 g%), total protein (2.37 gdL^?1) and globulin content (1.28 g dL^?1) during the pre‐challenge. Enzymes showed higher activities during post challenge (P<0.05). The result suggests that B. subtilis can be used effectively as a commercial product for use in aquaculture.     

The proteolytic digestive activity and growth during ontogeny of Parachromis dovii larvae (Pisces: Cichlidae) using two feeding protocols

The proteolytic digestive activity and growth of Parachromis dovii larvae during the ontogeny were evaluated in a recirculation system using two feeding strategies during a 28-day period. Larvae were reared using two feeding protocols (three replicates each): (A) Artemia nauplii (at satiation), fed from exogenous feeding [8 days after hatching (DAH)] until 15 DAH followed by nauplii substitution by formulated feed (20 % day^?1) until 20 DAH and then formulated feed until 28 DAH; (B) formulated feed (100 % BW daily) from exogenous feeding until 28 DAH. Levels of acid (pepsin type) and alkaline digestive proteases as well as growth and survival of larvae were measured along the feeding period. Survival was high and similar between treatments: 98.9 ± 0.0 for Artemia, 97.3 ± 0.0 % for formulated feed. The specific growth rate for length and weight was higher in larvae fed with Artemia nauplii than in larvae reared with formulated feed: 3.4 ± 0.1 versus 1.8 ± 0.1 % day^?1 for body length (P = 0.009) and 12.2 ± 0.1 versus 6.5 ± 0.3 % day^?1 for body weight (P = 0.002). The acid and alkaline proteolytic activity was detected, in both treatments, from the beginning of the experiment, at 8 DAH. The total enzymatic activity (U larva^?1) for acid and alkaline proteases was higher in larvae reared with Artemia after 12 DAH, whereas the specific enzymatic activity was similar for both enzyme types in the two treatments. The results suggest that P. dovii larvae were capable to digest formulated diets from the beginning of exogenous feeding and that they could be reared with formulated feeds. However, the formulated feed used should be nutritionally improved because of the poor growth obtained in this research.     

Use of different microbial probiotics in the diet of rohu,Labeo rohita fingerlings: effects on growth,nutrient digestibility and retention,digestive enzyme activities and intestinal microflora

Six iso‐nitrogenous (350 g protein kg^?1) and iso‐caloric (4100 kcal kg^?1) diets with or without probiotics supplementation namely T₁ (Basal feed (BF) without probiotics; control), T₂ (BF + Bacillus subtilis and Lactococcus lactis), T₃ (BF + L. lactis and Saccharomyces cerevisiae), T₄ (BF + B. subtilis and S. cerevisiae), T₅ (BF + B. subtilis, L. lactis and S. cerevisiae) and T₆ (BF + heat‐killed bacteria of B. subtilis, L. lactis and S. cerevisiae) were fed to Labeo rohita fingerlings (6.0 ± 0.06 g) for 60 days in triplicate tanks (30 fish per tank). In all probiotic‐supplemented diets, the probiotic concentration was maintained at 10¹¹ cfu kg^?1 feed. After 60 days of culture, the fish fed combination of three probiotics at equal proportion (T₅) had higher (P < 0.05) growth, protein efficiency ratio, nutrient retention and digestibility and lower (P > 0.05) feed conversion ratio over other treatment groups. Total heterotrophic bacterial population in intestine was drastically reduced on 15th and 30th days of sampling than the initial value (0 day of sampling) for T₃, T₄ and T₅ groups. Except T₆, the gut colonization of respective probiotics, which were supplemented through the diets, was also increased up to 30 days of culture of fish and thereafter remained constant.     

Demographic and competition studies on <Emphasis Type="Italic">Brachionus ibericus</Emphasis> and <Emphasis Type="Italic">Proales similis</Emphasis> in relation to salinity and algal (<Emphasis Type="Italic">Nannochloropsis oculata</Emphasis>) density

We isolated the rotifers Brachionus ibericus and Proales similis from the sediment of shrimp tanks and studied their individual demographic characters and competition between them at two food levels (0.25?×?10⁶, 1.00?×?10⁶ cells ml^?1 of Nannochloropsis oculata at 25 °C) and salinities ranging from 10 to 30‰. Our hypothesis was that growth rates would be higher with increasing food levels and salinities. Observations were taken twice a day for life table studies and daily once for population growth experiments. Using survivorship and fecundity data, we derived various life history variables. Although the average life span (7.6?±?0.4 days) and gross reproductive rate (33.8?±?2.9 neonate female^?1 day^?1) of B. ibericus were higher than those of P. similis (average life span 5.4?±?0.6 days and gross reproductive rate 13.0?±?0.6 neonate female^?1 day^?1), the population growth experiments showed that P. similis had higher r values (0.32?±?0.005 day^?1) than B. ibericus (0.23?±?0.002 day^?1) at 1.0?×?10⁶ cells ml^?1 of N. oculata. The rotifer P. similis was more adversely affected due to the presence of B. ibericus than vice versa. The data are important for developing techniques for a large-scale culture of these rotifers as food in aquaculture.     

The effects of feeding rates in juvenile Korean rockfish, (Sebastes schlegeli) reared at 17 °C and 20 °C water temperatures

Two feeding trials were conducted to determine the effects of feeding rates in juvenile Korean rockfish, (Sebastes schlegeli) reared at 17 and 20 °C water temperature. Fish averaging 5.5 ± 0.2 g (mean ± SD) at 17 °C and 5.5 ± 0.3 g (mean ± SD) at 20 °C water temperature were randomly distributed into 18 indoor tanks. At each water temperature, triplicate tanks were randomly assigned to one of six different feeding rates: 2.8, 3.8, 4.1, 4.4, 4.7 % and satiation (4.99 % BW day^?1) at 17 °C and 2.8, 3.8, 4.1, 4.4, 4.7 % and satiation (5.0 % BW day^?1) at 20 °C. After 4 weeks of feeding trial, weight gain (WG) and specific growth rate of fish fed groups at satiation and 4.7 % (BW day^?1) were significantly higher than those of fish fed groups at 2.8, 3.8 and 4.4 % (BW day^?1) in both 17 and 20 °C temperature. Feed efficiency and protein efficiency ratio of fish fed group at 2.8 % (BW day^?1) was significantly lower than those of fish fed groups at 3.8, 4.1, 4.4 and 4.7 % (BW day^?1) in both experiments. Hematocrit was significantly higher in fish fed group at 4.4 % (BW day^?1) at 17 °C, and there was no significant difference in hemoglobin content amongst all fish fed groups at 20 °C. Glutamic oxaloacetic transaminase and glutamic pyruvic transaminase of the fish fed group at 2.8 % (BW day^?1) were significantly higher than those of all other fish fed groups in both experiments. Broken line regression analysis of WG indicated that the optimum feeding rate of juvenile Korean rockfish was 4.48 % (BW day^?1) at 17 °C and 4.83 % (BW day^?1) at 20 °C. Therefore, these results indicated that the optimum feeding rate could be >4.1 % but <4.48 % at 17 °C and >4.4 % but <4.83 % at 20 °C. As we expected, current results have indicated that 5 g of juvenile Korean rockfish perform better at 17 °C than at 20 °C water temperature.     

Effects of traditional Chinese medicines (TCM) on the immune response of grass carp (Ctenopharyngodon idellus)

The use of traditional Chinese medicine (TCM) in aquaculture has been widely investigated. It is believed that herbal supplements could enhance the immunity and promote fish growth. The powdered TCM or its decoction can be easily applied in the form of feed supplements by direct incorporation into fish feeds and side effects are seldom found. In this study, Radix scutellaria (Rs) and Rhizoma coptidis (Rc) showed strong bacterial inhibition on Aeromonas hydrophila out of the sixteen selected herbal extracts. Four TCMs, R. scutellaria, R. coptidis, Herba andrographis (Ha), and Radix sophorae flavescentis (Rsf) were selected to form a compound formulation in the ratio of 1:1:2:3. A feeding experiment on grass carp was conducted with addition of the above TCM formulation (0.5, 1 and 2 % w/w) in the fish feed, and bactericidal activity, total protein and immunoglobulin in blood plasma, and nitroblue tetrazolium activity in the whole blood were measured. 2 % supplementation significantly improved (p < 0.05) the bactericidal activity and total immunoglobulin in the plasma after feeding for 21 days. Compared to control, 2 % TCM feeding groups showed a significantly lower mortality after A. hydrophila challenge at the end of experiment and the same result was archived in field trial. In the cost evaluation, the use of the TCM formulation in grass carp culture was insignificant and could be compensated by improved yield. TCM formulation contained different active ingredients including some immuno-stimulant and antimicrobial agents and therefore could be applicable to multiple diseases. Application of this TCM formulation would be a cost-effective and prophylactic approach for disease control in aquaculture, replacing the use of antibiotics for treating enteritis and even other general diseases.     

Synergy of microcapsule polysaccharides and Bacillus subtilis on the growth, immunity and resistance of sea cucumber Apostichopus japonicus against Vibrio splendidus infection

A 4-week feeding trial was conducted to determine the effects of different dietary supplements on the growth, immunity and resistance of sea cucumber Apostichopus japonicus against Vibrio splendidus infection. The control group was supplied with blank microcapsules, and Astragalus polysaccharide (APS) microcapsules, tuckahoe polysaccharide (TPS) microcapsules, (APS + TPS) microcapsules, (APS + TPS) microcapsules + Bacillus subtilis, were tested for effects. Coelomic fluid was collected at 7-day intervals to test activities of lysozyme (LSZ), superoxide dismutase (SOD), alkaline phosphatase (AKP), and complement 3 (C3) content. After the feeding trial, the specific growth rate of sea cucumbers fed a diet supplemented with (APS + TPS) microcapsules + B. subtilis was significantly increased (P < 0.05); activities of LSZ, SOD, AKP and C3 content were significantly higher than in other groups (P < 0.05). The challenge test showed that the cumulative mortality of sea cucumbers fed a diet supplemented with (APS + TPS) microcapsules + B. subtilis reduced significantly (P < 0.05). In conclusion, dietary combinations of (APS + TPS) microcapsules + B. subtilis has a potential for use in diet formulations for sea cucumbers to significantly increase growth, immunity and disease resistance against V. splendidus infection.     

Hemato-immunological responses of Heros severus fed diets supplemented with different levels of Dunaliella salina

In this study, the effects of oral administration of different levels of Dunaliella salina (a natural β-carotene source) on growth parameters, immunological and hematological indices, as well as skin carotenoids, of Heros severus were investigated. One hundred and eighty H. severus weighing 27 ± 0.5 g were divided randomly into four groups in triplicate (15 fish in each replicate). Groups 1–4 received food supplemented with 0, 50, 100 and 200 mg kg^?1 D. salina powder, respectively. After 6 weeks, the growth parameters were compared among the groups. Blood samples were taken from each group, and hematological parameters including red blood cell count (RBC), white blood cell count (WBC), hematocrit (PCV), hemoglobin (Hb) and immunological indices (serum and mucus lysozyme and bactericidal activity, resistance against Aeromonas hydrophila infection) as well as carotenoid content of skin were evaluated. Results showed that some growth indices increased significantly in fish fed with 100 and 200 mg kg^?1 D. salina-supplemented food (P < 0.05). Although serum lysozyme activity was increased in fish fed with food supplemented with 100 and 200 mg kg^?1 D. salina (P < 0.05), no significant change was observed in serum and mucus bactericidal activity and mucus lysozyme activity among the groups (P > 0.05). Most of the hematological parameters such as WBC, RBC, PCV and Hb significantly increased in D. salina-treated fish compared with controls (P < 0.05). Mortality induced after challenge with A. hydrophila in 200 mg kg^?1 D. salina-treated fish was 36.67 %, which significantly decreased compared with control (P < 0.05). Skin carotenoid content in all D. salina treatments was statistically higher than that of control (P < 0.05). Conclusively, D. salina as a food additive can affect positively the growth, immunological and hematological parameters of H. severus.     

<Emphasis Type="Italic">Virgibacillus proomii</Emphasis> and <Emphasis Type="Italic">Bacillus mojavensis</Emphasis> as probiotics in sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) larvae: effects on growth performance and digestive enzyme activities

This study examined the effects of two probiotics (Virgibacillus proomii and Bacillus mojavensis) on the digestive enzyme activity, survival and growth of Dicentrarchus labrax at various ontogenetic stages in three separate experiments. These probiotics were incorporated as single or mixed into fish feed for a period of 60 days. The growth parameters, proximate composition of whole body, digestive enzymes and gut microbiology were monitored at regular. The increments in length and weight and the survival were significantly higher (P < 0.05), and the values of food conversions were significantly lower (P < 0.05) in fishes fed the probiotic. The administration of V. proomii and B. mojavensis in diet resulted in an increase (P > 0.05) in body ash and protein content and in the specific activity of phosphatase alkaline and amylase in the digestive tract of all the fishes. V. proomii and B. mojavensis persisted in the fish intestine and in the feed in high numbers during the feeding period (group 1: 5.8 × 10⁴ CFU/ml, group 2: 9.6 × 10⁴ CFU/ml, and group 3: 9.8 × 10⁴ CFU/ml day 60). The two probiotics V. proomii and B. mojavensis were adequate for improved growth performance and survival and for healthy gut microenvironment of the host.