Effect of dietary chitosan on growth performance,hematological parameters,intestinal histology and stress resistance of Caspian kutum (<Emphasis Type="Italic">Rutilus frisii kutum</Emphasis> Kamenskii, 1901) fingerlings

This study was conducted to investigate the effects of dietary chitosan on growth performance, hematological parameters, intestinal histology, stress resistance and body composition in the Caspian kutum (Rutilus frisii kutum, Kamenskii, 1901) fingerlings. Fish (1.7 ± 0.15 g) were fed diets containing chitosan at different levels (0, 0.25, 0.5, 1 and 2 g kg^?1 diet) for a period of 60 days. Results showed that the feed conversion ratio significantly decreased in fish fed diet containing 1 g kg^?1 of chitosan compared to the other groups (P < 0.05), but there were no significant differences between treatments in terms of specific growth rate and condition factor (P > 0.05). Leukocyte increased in fish fed diet containing 2 g kg^?1 of chitosan compared to the other groups (P < 0.05). Lymphocytes, eosinophils and neutrophils did not significantly change among dietary treatments (P > 0.05). Also, the chitosan supplementation did not affect the whole-fish body composition (P > 0.05). Light microscopy demonstrated that the intestinal villus length increased in fish fed diet containing 1 g kg^?1 of chitosan compared to control group (P < 0.05). While 11 and 13 ‰ salinity and 30 °C thermal stress had no effect, 1 g kg^?1 of chitosan (P < 0.05) showed highest survival rate (70 %) in 34 °C thermal stress. The results showed that chitosan in the diet of the Caspian kutum fingerlings could improve feed conversion ratio, the nonspecific defense mechanisms and resistance to some of the environmental stresses.     

Dietary protein and carbohydrate requirement of juvenile Hawaiian limpet (<Emphasis Type="Italic">Cellana sandwicensis</Emphasis> Pease, 1861) fed practical diet

This study was conducted to evaluate dietary protein and carbohydrate requirement of juvenile Hawaiian limpets Cellana sandwincensis. A total of 64 juvenile limpets (3.12 ± 0.86 g) were fed five different dietary protein levels ranging from 270 to 470 g kg^?1 for 90 days. Carbohydrate and lipid levels were held constant at 180 and 49.7 g kg^?1, respectively. Weight gain and growth rates of the animals did not differ significantly (P > 0.05) among the protein levels ranging from 270 g kg^?1 (0.30 % day^?1) to 470 g kg^?1 (0.23 % day^?1). Next, opihi were fed four diets with protein levels from 210 to 500 g kg^?1 with a constant carbohydrate level at 120 g kg^?1. Weight gain and specific growth rates of opihi increased with increasing dietary protein from 210 to 350 g kg^?1, and significantly (P < 0.05) decreased at the 500 g kg^?1 diet. Highest weight gain, growth rates, and protein efficiency ratio were achieved at 350 g kg^?1. Elevated carbohydrate levels (180–370 g kg^?1) produced a significant difference (P < 0.05) in growth. The fastest growth rates of animals were obtained with 270 g kg^?1 (0.27 % day^?1) and 320 g kg^?1 (0.26 % day^?1). The weight gain of animals fed 180 and 370 g kg^?1 carbohydrate diets were significantly (P < 0.05) lower than those of animals fed 270 and 320 g kg^?1. We conclude that about 350 g kg^?1 protein and 320 g kg^?1 carbohydrate levels could be used for opihi.     

Modulation of appetite,lipid and glucose metabolism of juvenile grass carp (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>) by different dietary protein levels

This study was undertaken to explore the systemic metabolic strategies of juvenile grass carp (Ctenopharyngodon idellus) to maintain growth when fed with different dietary protein levels. The optimal growth group and two growing discomfort groups were selected through the basic data, to explain the growth difference from appetite regulation and lipid and glucose metabolism perspective. Three experimental diets were formulated with three dietary protein levels at 200.3, 296.1 and 442.9 g kg^?1, named P1, P2 and P3, respectively. Juvenile grass carp (initial body weight 12.28 ± 0.14 g) were fed with three diets with 3 replications per dietary treatment in an indoor recirculation system for an 8-week feeding trial. Fish fed with diet P2 dietary group showed significantly higher WG, SGR, FI and PER than other groups. Compared with other groups, mRNA expressions of NPY, Y8a and Y8b in fish fed with P2 significantly down-regulated, while the expressions of CCK and CART in fish fed with P3 significantly down-regulated (P < 0.05). With increasing dietary protein levels, G6Pase, GK, PK and PEPCK were all significantly inhibited (P < 0.05). For lipid metabolism, the mRNA expression of ACC in P1 dietary group was significantly higher than P3 dietary group; besides, LPL expression in P3 group was significantly higher than other two groups (P < 0.05). PPARα expression in P2 was significantly lower than other groups (P < 0.05). These results suggested that grass carp fed with P2 (296.1 g kg^?1 protein level) showed highest weight gain, contributed to more balanced nutrient metabolism and appetite regulation. Too high dietary protein (442.9 g kg^?1) should be avoided because it induced lowest PER, body lipid and liver lipid, and inhibited glucose and lipid metabolism in juvenile grass carp.     

Effect of stocking density on growth performance,serum biochemical parameters,and muscle texture properties of genetically improved farm tilapia, <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

The objective of this study was to assess the effects of stocking density on growth performance, serum biochemical parameters, and muscle texture properties of genetically improved farmed tilapia (Oreochromis niloticus, GIFT). Juvenile GIFT with an average initial weight of 12.54?±?0.45 g (mean?±?SD) were randomly stocked in 16 tanks (80 L) in a recirculation aquaculture system at four densities of 10 (D1), 20 (D2), 30 (D3), and 40 (D4) fish per tank for 56 days, with quadruplicate for each density. There were no significant differences in water temperature among the four treatments (P?>?0.05). D4 had the significantly lowest dissolved oxygen content (5.52 vs 5.69–6.09 mg L^?1) (P?>?0.05) and pH (6.63 vs 6.87–7.20) (P?<?0.05). NO₂-N and NH₄-N concentrations significantly increased with increasing stocking density (P?<?0.05). Weight gain (WG) and specific growth rates (SGR) decreased with increasing stocking density. The lowest WG (617.20 vs 660.45–747.06%), SGR (3.52 vs 3.62–3.81% day^?1), and highest feed conversion ratio (1.68 vs 1.53–1.58) were observed in D4. Fish at D4 had significantly lower condition factor (3.11 vs 3.29–3.37%) and survival rate (91.25 vs 97.50%) than those from D1 and D2 (P?<?0.05). With increasing stocking density, serum total cholesterol, triglyceride, and total protein concentrations decreased (P <?0.05) and aspartate aminotransferase and alanine aminotransferase activities increased (P <?0.05). D4 fish had higher moisture content (78.80 vs 76.97%) and lower crude protein content (18.14 vs 19.39%) in muscle than D1 fish (P?<?0.05). Compared to D1 and D2, D3 and D4 had lower muscle hardness (1271.54–1294.07 vs 1465.12–1485.65 g), springiness (0.62–0.65 vs 0.70–0.72), gumminess (857.33–885.32 vs 1058.82–1079.28 g), and chewiness (533.04–577.09 vs 757.53–775.69 g) (P <?0.05). High stocking density resulted in growth inhibition, declines in flesh quality, and disturbance to several serum biochemical parameters.     

Effects of exogenous non-starch polysaccharide-degrading enzymes in diets containing <Emphasis Type="Italic">Gracilaria lemaneiformis</Emphasis> on white-spotted snapper <Emphasis Type="Italic">Lutjanus stellatus</Emphasis> Akazaki

A feeding experiment was conducted to investigate the effects of exogenous non-starch polysaccharide (NSP)-degrading enzymes in diets containing Gracilaria lemaneiformis (GL) on growth performance and digestive enzyme activities of white-spotted snapper Lutjanus stellatus Akazaki (initial mass 8.0 ± 0.1 g). A basal diet (D0) containing a mixed protein source (fish meal, soybean meal and GL meal) was used as the control. Two diets supplemented with 0.5 g (D1) and 1 g (D2) exogenous NSP-degrading enzymes per kilogram of diet were formulated. Each diet was assigned to triplicate groups of fish in a total of nine floating sea cages (270 fish, 30 fish per cage). After a 60-day feeding trail, significantly higher weight gain, specific growth ratio and feed efficiency ratio were observed in fish fed D2 diet compared to those of control (P < 0.05). Body lipid, moisture and ash contents were not significantly affected by NSP-degrading enzyme supplementation, but significantly higher protein content was noticed in fish fed D2 diet compared to that of control (P < 0.05). The amylase activity in the stomach and intestine was significantly higher in fish fed D2 diet (P < 0.05), but no significant differences were observed in pepsin and lipase activities in the stomach or trypsin and lipase activities in the intestine between all treatments. The results suggested that addition of 1 g kg^?1 NSP-degrading enzymes in diet could efficiently improve seaweed feed utilization and growth performance of white-spotted snapper fish.     

Stimulatory effect of dietary taurine on growth performance,digestive enzymes activity,antioxidant capacity,and tolerance of common carp, <Emphasis Type="Italic">Cyprinus carpio</Emphasis> L., fry to salinity stress

The present study was carried out to evaluate the effect of dietary taurine (Tau) on performance, digestive enzymes, antioxidant activity, and resistance of common carp, Cyprinus carpio L., fry to salinity stress. Fish (0.97?±?0.033 g) were fed on different taurine levels of 0.0 (control), 5, 10, 15, or 20 g/kg diet up to satiation twice daily for 8 weeks. At the end of the feeding trial, fish were stressed by exposure to 10 ppt salinity for 3 days during which fish mortality was observed. Fish performance was significantly (P?<?0.05) improved by dietary taurine up to 15 g Tau/kg diet after which fish growth and feed intake were almost the same. Also, taurine supplementation significantly (P?<?0.05) elevated activities of intestinal amylase, lipase, and protease resulting in an improving in feed intake giving better performance. Furthermore, Tau-stimulated antioxidant activity of common carp was observed in a dose-related manner, where activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were significantly (P?<?0.05) higher, but malondialdehyde (MDA) value was significantly (P?<?0.05) lower in Tau-fed fish groups than those fed the control diet. In salinity stress experiment, highest survival rate was observed at fish fed Tau-supplemented diets without significant (P?>?0.05) differences over fish fed the control diet. It appears that taurine could be used as a feed supplement to confer better growth and health of common carp fry with optimal level of 15 g/kg diet.     

Effects of nucleotides on growth performance,immune response,disease resistance and intestinal morphology in shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> fed with a low fish meal diet

Nucleotides (NT) are widely used as an immunostimulant in aquaculture. A 10-week feeding trial was conducted to evaluate the effects of dietary NT on the growth performance, immune response, disease resistance and intestinal morphology of the Pacific white shrimp Litopenaeus vannamei (initial mean body weight: 0.39 ± 0.00 g). Five isolipidic (about 7 % crude lipid) and isonitrogenous (about 39 % crude protein) practical diets were supplemented with graded levels of NT (0, 60, 90, 120 and 1200 mg kg^?1), respectively. These diets were named as N1, N2, N3, N4 and N5. Each diet was randomly fed to six tanks of shrimps, and each tank contained 40 shrimps. The shrimps were fed four times daily (07:00, 11:00, 16:00 and 21:00 h). The results showed that there were no significant differences in survival, final weight, specific growth rate, feed intake, feed conversion ratio and whole-body compositions among all the treatments (P > 0.05). Shrimps fed the control diet had the significantly lowest activities of superoxide dismutase, total nitric oxide synthase and lysozyme (P < 0.05). However, those parameters increased when dietary NT increased from 60 to 120 mg kg^?1 and decreased as inclusion level increased to 1200 mg kg^?1 (P < 0.05). The cumulative mortality of the shrimps challenged with Vibrio parahaemolyticus was significantly higher in the treatments without dietary NT supplementation than those in treatments with dietary NT (≥90 mg kg^?1) (P < 0.05). The jejunum wall thickness in shrimps fed the control diet was significantly lower than that in the treatments of N2, N3 and N4 (P < 0.05). For the villus height, the highest value (49.29 µm) was found in N3. In summary, the present study showed that 90 mg kg^?1 of dietary NT is the optimum dietary level for good gut health, immune response and disease resistance of Pacific white shrimp fed a diet with 18 % fish meal.     

The effects of starvation on fast-start escape and constant acceleration swimming performance in rose bitterling (<Emphasis Type="Italic">Rhodeus ocellatus</Emphasis>) at two acclimation temperatures

To investigate the effects of starvation and acclimation temperature on the escape ability of juvenile rose bitterling (Rhodeus ocellatus), we measured the fast-start escape and constant acceleration swimming performance of fish fasted for 0 (control), 1 and 2 weeks and half-lethal periods (6 or 4 weeks) at two temperatures (15 and 25 °C). Fish acclimated at a high temperature exhibited shorter response latency (R), higher maximum linear velocity (V _max) and longer escape distance during escape movement (D _120ms) than those at the low temperature. Starvation resulted in a significant decrease in V _max and D _120ms at either low or high temperature and a significant increase in R at only the high temperature in the half-lethal period groups (P < 0.05). The relationship between V _max (Y, m s^?1) and starvation time (X, week) was Y ₁₅ = ?0.062X + 1.568 (r = ?0.665, n = 36, P < 0.001) at low temperature and Y ₂₅ = ?0.091X + 1.755 (r = ?0.391, n = 40, P = 0.013) at high temperature. The relationship between U _cat (Y, cm s^?1) and starvation time (X, week) was Y ₁₅ = ?1.649X + 55.418 (r = ?0.398, n = 34, P = 0.020) at low temperature and Y ₂₅ = ?4.917X + 62.916 (r = ?0.793, n = 33, P < 0.001) at high temperature. The slopes of equations showed a significant difference between low and high temperature (F _1,63 = 9.688, P = 0.003), which may be due to the different energy substrate utilization when faced with food deprivation at different temperatures.     

Effects of salinity,stocking density,and algal density on growth and survival of Iwagaki oyster <Emphasis Type="Italic">Crassostrea nippona</Emphasis> larvae

To determine the optimal salinity, stocking density, and algal density for hatchery culture of the Iwagaki oyster Crassostrea nippona larvae, three experiments with salinities of 14, 18, 22, 26, 30, and 34 practical salinity unit (PSU); stocking densities of 0.5, 1, 2, 4, 8, and 12 larvae ml^?1; and algal densities of 10, 20, 40, and 100?×?10³ cells ml^?1 were designed, which included the developmental stages from newly hatched D-larvae to pediveligers. Results showed that larval growth of C. nippona was the fastest at a salinity of 26 PSU, and when salinity was adjusted to a level that was lower or higher than this salinity, survival and growth rate of larvae declined (P <?0.05), resulting both in a decreased mean shell length and a high mortality. Larval growth decreased significantly with increasing stocking density. Larvae reared at 4 larvae ml^?1 had the smallest shell length (198.9 μm) and lowest survival rate (7.9%), whereas larvae reared at 0.5 larvae ml^?1 had the largest shell length (245 μm) and highest survival rate (66.3%) on day 13. And the shell length of larvae reared at 0.5 and 1 larvae ml^?1 was significantly (P?<?0.05) larger than the values in other treatments, except those reared at 2 larvae ml^?1 (P?>?0.05). When feeding the single-algal diet of Isochrysis galbana (clone T-ISO), the shell length of larvae increased markedly as the algal density was increased. Larvae reared at the highest algal density (100?×?10³ cells ml^?1) had the largest mean shell length; however, under the conditions of our experiment, there was no significant difference (P?>?0.05) in growth and survival rates between the treatments at algal densities of 40?×?10³ and 100?×?10³ cells ml^?1. For a large-scale culture, based on the results of this study, a salinity of 26 PSU, stocking density of 0.5–1 larvae ml^?1, and algal density of 40?×?10³ cells ml^?1 are recommended for an early development of C. nippona.     

Comparative evaluation of fermented and non-fermented de-oiled rice bran with or without exogenous enzymes supplementation in the diet of <Emphasis Type="Italic">Labeo rohita</Emphasis> (Hamilton, 1822)

A 60-day feeding trial was conducted to study the effect of exogenous enzymes (xylanase and phytase) supplementation in the non-fermented and fermented de-oiled rice bran (DORB)-based diet of Labeo rohita. Four test diets (T1-DORB-based diet, T2-fermented DORB-based diet, T3-phytase and xylanase supplemented DORB-based diet, and T4-phytase and xylanase supplemented fermented DORB-based diet) were formulated and fed to the respective groups. Test diets T3 and T4 were supplemented with 0.01% xylanase (16,000 U kg^?1) and 0.01% phytase (500 U kg^?1) enzymes. One hundred twenty juveniles of L. rohita, with an average weight 5.01?±?0.02 g, were stocked in 12 uniform size plastic rectangular tanks in triplicate with 10 fishes per tank following a completely randomized design (CRD). Exogenous enzyme supplementation to the T3 group significantly improved the growth performance of L. rohita (p?<?0.05). Fermented DORB fed groups registered significantly lower growth irrespective of the supplementation of exogenous enzymes. The carcass composition (except CP %), enzyme activities (except amylase activity), globulin, and A/G ratio did not vary significantly (p?>?0.05). Based on the results of the present study, it is concluded that exogenous enzyme supplementation significantly increases the growth of fish fed with DORB-based diet.     

Physiological response,blood chemistry profile and mucus secretion of red sea bream (<Emphasis Type="Italic">Pagrus major</Emphasis>) fed diets supplemented with <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> under low salinity stress

Environmental stressors caused by inadequate aquaculture management strategies suppress the immune response of fish and make them more susceptible to diseases. Therefore, efforts have been made to relieve stress in fish by using various functional feed additives in the diet, including probiotics. The present work evaluates the effects of Lactobacillus rhamnosus (LR) on physiological stress response, blood chemistry and mucus secretion of red sea bream (Pagrus major) under low salinity stress. Fish were fed four diets supplemented with LR at [0 (LR0), 1 × 10² (LR1), 1 × 10⁴ (LR2) and 1 × 10⁶ (LR3) cells g^?1] for 56 days. Before stress, blood cortisol, urea nitrogen (BUN) and total bilirubin (T-BIL) showed no significant difference (P > 0.05), whereas plasma glucose and triglyceride (TG) of fish-fed LR2 and LR3 diets were significantly lower (P < 0.05) than those of the other groups. Plasma total cholesterol (T-CHO) of fish-fed LR3 diet was significantly (P < 0.05) lower than that of the other groups. Furthermore, total plasma protein, mucus myeloperoxidase activity and the amount of mucus secretion were significantly enhanced in LR-supplemented groups when compared with the control group (P < 0.05). After the application of the low salinity stress test, plasma cortisol, glucose, T-CHO and TG contents in all groups showed an increased trend significantly (P < 0.01) compared to the fish before the stress challenge. However, plasma total protein and the amount of secreted mucus showed a decreased trend in all groups. On the other hand, BUN, T-BIL and mucus myeloperoxidase activity showed no significant difference after exposure to the low salinity stress (P > 0.05). In addition, the fish that received LR-supplemented diets showed significantly higher tolerance against low salinity stress than the fish-fed LR-free diet (P < 0.05). The physiological status and the detected immune responses, including total plasma protein and mucus myeloperoxidase activity in red sea bream, will provide a more comprehensive outlook of the effects of probiotics to relieve stress in fish.     

Substitution of soybean meal with fermented <Emphasis Type="Italic">Jatropha</Emphasis> kernel meal: effect on growth performance,body composition,and metabolic enzyme activity of <Emphasis Type="Italic">Labeo rohita</Emphasis>

A 60-day feeding trial was conducted to evaluate the nutritional potential of fermented Jatropha kernel meal (FJKM) in the diets of rohu, Labeo rohita, fingerlings. Four iso-nitrogenous (336 g kg^?1) and iso-energetic (20 MJ kg^?1) diets containing 0, 100, 200, and 300 g kg^?1 FJKM in replacement of 0, 33.3, 66.7, and 100% soybean meal protein, respectively, were prepared and designated as control, T1, T2, and T3, respectively. A total of 180 acclimatized fish (average weight 6.05?±?0.04 g) were equally distributed into four experimental groups in triplicates, and were fed their respective diets to satiation twice daily at 10:00 h and 18:00 h. Higher percent weight gain (%WG); specific growth rate (SGR); protein efficiency ratio (PER); protease, aspartate amino transferase (AST), and alanine amino transferase (ALT) activities; and lower feed conversion ratio (FCR) were recorded in T2 and T3 groups compared to other groups. Whereas hepatosomatic index (HSI), intestinal somatic index (ISI), and amylase, lactate dehydrogenase (LDH), malate dehydrogenase (MDH), superoxide dismutase (SOD), and catalase activities among the various dietary groups did not vary significantly (P?>?0.05). Whole body composition of fish analyzed at the end of the feeding trial exhibited significantly (P?<?0.05) higher ether extract and lower crude protein in the control group compared to the FJKM-fed groups. These results indicated that rohu fingerlings can efficiently utilize FJKM without any detrimental effect on growth performance, nutrient utilization, and metabolic response.     

In vitro and in vivo evaluation of probiotic properties of <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> in Kenyi cichlid, <Emphasis Type="Italic">Maylandia lombardoi</Emphasis>

In this study, Enterobacter cloacae bacterium was isolated from curd and its antibacterial potential against the pathogen Plesiomonas shigelloides was evaluated using the freshwater ornamental fish Kenyi cichlid (Maylandia lombardoi). Among the bacterial isolates, E. cloacae exhibited tolerance to acidic pH 2 and demonstrated the highest antibacterial activity against P. shigelloides in various in vitro assays. Dietary supplementation of E. cloacae with prebiotic supplement 2% mannan oligosaccharide improved the growth performance and reduced the toxic metabolites such as nitrite in culture tank water of Kenyi cichlid. In vivo application of E. cloacae with mannan oligosaccharide significantly elevated (p?<?0.05) white blood cell counts (88.47?±?2.15 10³ mm^?3) and respiratory burst activity (0.243?±?0.007) of Kenyi cichlid when challenged with P. shigelloides. Intestinal morphology of Kenyi cichlid fish treated with probiotic and prebiotic combinations showed improved intestinal architectures. The present findings confirm that the isolated bacterium E. cloacae is a potential probiotic and can be used effectively to prevent infection of P. shigelloides in freshwater ornamental fish culture.     

Corn gluten meal in pacu <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> diets: effects on growth,haematology, and meat quality

Corn gluten meal (CGM) has a high protein content and absence of antinutrients. However, it has high levels of carotenoids that can cause a yellowing of fish fillets and impair further commercialisation. Graded levels of CGM were incorporated in pacu diets to replace soybean meal (SM) protein. The experimental diets were formulated to be isonitrogenous (220 g kg^?1 digestible protein) and isoenergetic (13.4 MJ kg^?1 digestible energy) with 0, 25, 50, 75, and 100 % replacement of SM protein by CGM protein over a 60-day period. One hundred fish (32.79 ± 3.43 g) were randomly distributed in 20 experimental cages (70 L) placed within five (1000 L) aquaria with continuous water renewal. The experimental design was completely randomised with five treatments and four replicates. A quadratic effect (P < 0.05) was observed for fillet and carcass yields, feed conversion ratio, weight gain, and specific growth rate, with the optimum values for replacement of SM protein by CGM protein estimated as 21.95, 29.13, 30.94, 37.71, and 38.75 %, respectively. There were no differences (P > 0.05) for blood parameters, proximal composition, water-holding capacity, pH, lipid oxidation, and a* value of fillets. The L* and b* values of fillets showed differences (P < 0.05). Replacement between 21.95 and 38.75 % of SM protein by CGM protein (5.37 and 9.48 % of inclusion in the diet) improves the growth performance and body yield of pacu juveniles without altering blood parameters or affecting the fillet quality.     

Inorganic nitrogen control,growth, and immunophysiological response of <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis> (Boone, 1931) in a biofloc system and in clear water with or without commercial probiotic

A 5-week feeding trial was conducted to evaluate the effects of biofloc in situ and commercial probiotic supplementation on white shrimp (1.87?±?0.03 g) inorganic nitrogen control, growth, and immunophysiological response. For this purpose, four treatments were conducted: clear water with no probiotic application (CW), clear water with probiotic application (CW+P), biofloc with no probiotic application (FLOC), and biofloc with probiotic application (BFT+P); each group had three replicates. Growth parameters (final body weight, daily weight gain, specific growth rate) were significantly higher in the two biofloc systems (P?<?0.05), and the FLOC and FLOC+P group did not have a significant difference (P?>?0.05). The immune responses (total hemocyte count, complement component protein, and lysozyme) and antioxidant status (glutathione, catalase) in the CW+P, FLOC, and FLOC+P groups were increased significantly at the end of the experiment compared with the CW group (P?<?0.05), and the FLOC and FLOC+P groups did not have a significant difference (P?>?0.05). Results of a 10-day Vibrio harveyi challenge test show that the survival rates in CW+P, FLOC, and FLOC+P groups were significantly higher (P?<?0.05), and the FLOC and FLOC+P groups did not have a significant difference (P?>?0.05). These results suggest that probiotic addition in the biofloc system had little advantage, but probiotics can improve the immune status of the shrimp in the clear water system. Further, cost-effectiveness analysis showed that the biofloc system was an efficient and economical option for the production of white shrimp.     

Effect of dietary protein levels and feeding rates on the growth and health status of juvenile genetically improved farmed tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

A 2 × 3 factorial study was conducted to investigate the effects of dietary protein levels (DPLs) and feeding rates (FRs) on the growth and health status of juvenile genetically improved farmed tilapia (GIFT), Oreochromis niloticus. Triplicate tanks of fish (initial weight 15.87 ± 0.11 g) were fed diets containing 25 or 35% protein at rates of 3, 5, or 7% body weight per day (BW day^?1) for 8 weeks. At the end of the feeding trial, the results showed that fish growth (final mean weight 34.61–81.07 g) and condition factor (3.39–4.45 g cm^?3) increased with the DPLs and FRs. Feed efficiency (FE, 0.48–0.88) increased as DPLs increased but decreased as FRs increased; the opposite trend was observed for feed cost (FC, 3.24–5.82 CHN Yuan kg^?1) and hepatosomatic index (0.98–2.33%). Apparent protein retention efficiency (APRE, 23.92–38.78%) was reduced by high FR. A 35% protein diet resulted in higher (P < 0.05) FE and APRE, and lower (P < 0.05) FC at 5% BW day^?1 than those at 7% BW day^?1. As FRs increased, lipid contents of the hepatopancreas, viscera, muscle, and eviscerated body increased, while moisture contents of hepatopancreas and viscera decreased. All serum biochemical parameters, including glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase activity and levels of creatinine, glucose, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, total cholesterol, triglyceride, and total protein were unaffected by DPL or FR (P > 0.05), except urea nitrogen levels, which were affected by DPLs (P < 0.05). Moreover, the size of hepatocytes and the area ratio of hepatocyte vacuoles were enlarged (P < 0.05), whereas the area ratios of the nucleus and cytoplasm were reduced (P < 0.05) with increasing FRs. These results suggested that the optimal feeding strategy for juvenile GIFT is 35% protein diet at 5% BW day^?1.     

Effect of nitrite exposure on oxygen-carrying capacity and gene expression of <Emphasis Type="Italic">NF-κB</Emphasis>/<Emphasis Type="Italic">HIF-1α</Emphasis> pathway in gill of bighead carp (<Emphasis Type="Italic">Aristichthys nobilis</Emphasis>)

Nitrite (NO₂^?) contamination of water can severely impact the health of aquatic life and is a major concern for commercial aquaculture. In order to study the effect of nitrite on Aristichthys nobilis, we investigated the oxygen-carrying capacity, NF-κB/HIF-1α pathway, and the gill tissue structure under nitrite stress. In the current study, bighead carp (initial weight 180.05?±?0.092 g) were exposed to nitrite (48.634 mg/L) for 96 h and then for 96 h recovery test. After nitrite exposure for 6 h, hypoxia-inducible factor-1α (HIF-1α) and nuclear factor-κB (NF-κB) mRNA expression increased significantly in the gill of bighead carp (P?<?0.05). After nitrite exposure for 12 h, hemoglobin (Hb) and methemoglobin reductase (MHBR) content in blood decreased significantly (P?<?0.05); TLR4 mRNA expression increased significantly (P?<?0.05). After nitrite exposure for 24 h, methemoglobin (MetHb) content increased significantly (P?<?0.05). After recovery test, all the indicators except TLR4 mRNA expression level recovered to initial level. In conclusion, nitrite exposure can affect hemoglobin dynamics, as oxidization of nitrite by hemoglobin results in the reduction of Hb to MetHb leading to hypoxia and nitrite exposure can also result into gill tissue damage. In the face of nitrite exposure, NF-κB and HIF-1α mRNA expression level increased immediately to protect the body from oxidative damage and eased hypoxic condition caused by nitrite. It was also observed that nitrite damage is recoverable in Aristichthys nobilis, but it may be need more than 96 h.     

Dietary ascorbic acid influences the intestinal morphology and hematology of hybrid sorubim catfish (<Emphasis Type="Italic">Pseudoplatystoma reticulatum</Emphasis> × <Emphasis Type="Italic">P. corruscans</Emphasis>)

This study evaluated the effect of graded levels of dietary ascorbic acid (AA) (12.47, 20.27, 115.44, 475.50, 737.72, and 850.70 mg kg^?1) on growth, hematology, intestinal morphometry, and phagocyte activity of hybrid sorubim Pseudoplatystoma reticulatum × Pseudoplatystoma corruscans. Fish (n = 420, 14.57 ± 2.71 g, 15.11 ± 0.90 cm) were distributed in 30 polyethylene tanks (80 l) (5 replicates per treatment with 14 fish per tank) and fed for 45 days. Dietary treatment did not have a significant effect on growth metrics (P > 0.05). Fish fed 737.72 mg AA kg^?1 had a higher villi height (289.80 ± 19.96 μm) (P < 0.05) than fish fed 850.70 mg AA kg^?1 (245.4 ± 18.25 μm). Hemoglobin in fish fed 850.70 mg AA kg^?1 (5.34 ± 0.96 g dl^?1) was higher (P < 0.05) than fish fed 12.47 mg AA kg^?1 (3.42 ± 0.55 g dl^?1) and 20.27 mg AA kg^?1 (3.06 ± 1.26 g dl^?1). The erythrocyte number of hybrid sorubim fed 115.40 mg AA kg^?1 (1.73 ± 0.27 × 10⁶ μl^?1) and 475.50 mg AA kg^?1 (1.70 ± 0.28 × 10⁶ μl^?1) were higher (P < 0.05) than in those fed diets containing 20.27 mg AA kg^?1 (1.11 ± 0.34 × 10⁶ μl^?1). There was no significant effect (P > 0.05) of dietary AA on leukocyte and thrombocyte and on phagocyte activity and phagocyte index. Inclusion of AA in feed seems to increase the integrity of the intestinal mucosa and stimulate erythropoiesis in hybrid sorubim catfish.     

Dietary docosahexaenoic acid decreased lipid accumulation via inducing adipocytes apoptosis of grass carp, <Emphasis Type="Italic">Ctenopharygodon idella</Emphasis>

The purpose of this study was to explore the mechanism of by which docosahexaenoic acid (DHA) inhibit the accumulation of adipose tissue lipid in grass carp (Ctenopharyngodon idella). We therefore designed two semi-purified diets, namely DHA-free (control) and DHA-supplemented, and fed them to grass carp (22.19 ± 1.76 g) for 3 and 6 weeks. DHA supplementation led to a significantly lower intraperitoneal fat index (IPFI) than that in the control group by reducing the number of adipocytes but significantly higher adipocyte size (P < 0.05). In the intraperitoneal adipose tissue, the DHA-fed group showed significantly higher peroxisome proliferator-activated receptor (PPAR)γ, CCAAT enhancer-binding protein (C/EBP)α, and sterol regulatory element-binding protein (SREBP)1c mRNA expression levels at both 3 and 6 weeks (P < 0.05). However, the ratio of the expression levels of B cell leukemia 2 (Bcl-2) and Bcl-2-associated X protein (Bax) was significantly lower in the DHA-fed group than in the control group (P < 0.05), and the protein expression levels of the apoptosis-related proteins caspase 3, caspase 8, and caspase 9 were also significantly higher (P < 0.05). Overall, although DHA promotes lipid synthesis, it is more likely that DHA could suppress the lipid accumulation in adipocytes of grass carp by inducing adipocyte apoptosis.     

The changes in cortisol and expression of immune genes of GIFT tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> (L.) at different rearing densities under <Emphasis Type="Italic">Streptococcus iniae</Emphasis> infection

The present study was conducted to investigate the effects of stocking density on serum cortisol (COR) levels and expression of immune genes in the head kidney of juvenile GIFT tilapia (Oreochromis niloticus) after Streptococcus iniae (S. iniae) infection. Juveniles (2700) were distributed into 30 tanks at five stocking densities (150, 300, 450, 600 and 750 g/m³), and each treatment had six replicates. After a 45-day feeding trial, a S. iniae challenge study was conducted for 96 h. Differences in mortality, serum COR and gene expression of C-type lysozyme, heat-shock protein 70 (HSP70), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) were analyzed. The levels of serum COR at all infected treatments were significantly higher compared with the levels of pre-infection (P < 0.05). At 96 h post-infection, serum COR levels of high stocking densities (600 and 750 g/m³) were significantly higher than those of low-density groups (P < 0.05); the mRNA levels of C-type lysozyme, HSP70, IL-1β, TNF-α and IFN-γ were significantly lower in tilapia reared at 600 g/m³ group than in those reared at 300 g/m³ group (P < 0.05). Fish reared in high-density treatments (600 and 750 g/m³) exhibited significantly higher post-challenge mortality (43.3 and 40.0 %) at 96 h following S. iniae infection, and it was significantly higher than other groups (P < 0.05). The data suggest that when living in high stocking density, GIFT tilapia showed decrease in immune capability, together with increased serum COR and susceptibility to S. iniae.