Spawning of female Black tiger shrimp (Penaeus monodon) is not impacted by muscle injection of dsRNA targeted to gill‐associated virus

The ability of domesticated Penaeus monodon, Black Tiger shrimp, to spawn following tail‐muscle injection of dsRNA was examined. Ablated domesticated female broodstock infected subclinically with gill‐associated virus (GAV) were injected with saline or a cocktail of five‐dsRNAs targeting different regions in the GAV ORF1a/1b gene. To track changes in GAV infection loads, TaqMan real‐time PCR was used to quantify mean viral RNA amounts in each of three pleopod clips collected at the time of injection (Day 0) and either immediately after a female spawned or on Day 11 when the trial was terminated. Over the trial, 4 of 19 (21%) saline‐injected shrimp spawned and 12 of 25 (48%) dsRNA‐injected shrimp spawned, with one spawning twice. Egg numbers varied from 25 600 to 459 800 for the saline‐injected shrimp and from 4900 to 213 900 for the dsRNA‐injected shrimp. Of these, one of the four egg batches hatched from saline‐injected shrimp and 9 of the 13 egg batches hatched from dsRNA‐injected shrimp. While variable, egg numbers and hatch rates recorded were typical of those obtained from domesticated broodstock at the commercial hatchery and particularly among females previously spawned. Mean GAV RNA amounts detected in pleopod samples increased in five of the eight saline‐injected shrimp tested by 1.6–227.4‐fold and decreased in 12 of the 15 ds‐RNA‐injected shrimp tested by ?1.1 to ?45.1‐fold. The study demonstrated that tail‐muscle injection of GAV‐specific dsRNA does not adversely impact the ability of P. monodon to spawn.     

Optimum dietary protein‐to‐energy ratio for juvenile whiteleg shrimp,Litopenaeus vannamei,reared in a biofloc system

An 8‐week feeding trial was conducted to evaluate the optimum dietary protein‐to‐energy (P/E) ratio in juvenile whiteleg shrimp, Litopenaeus vannamei. Six diets were formulated with two protein levels (30% and 35%) and three digestible energy levels (16, 17.5 and 19 kJ/g diet) at each protein level (₃₀P₁₆, ₃₀P_17.5, ₃₀P₁₉, ₃₅P₁₆, ₃₅P_17.5 and ₃₅P₁₉). Fifty shrimp averaging 0.97 ± 0.03g (Mean ± SD) were randomly distributed in biofloc tanks and fed one of the experimental diets. Weight gain (WG), specific growth rate (SGR) and feed efficiency (FE) of shrimp fed the ₃₅P_17.5 diet were significantly higher than those of shrimp fed ₃₀P₁₆, ₃₀P_17.5 and ₃₀P₁₉ diets (p < .05). Results for non‐specific immune responses showed that diet ₃₅P_17.5 caused higher lysozyme activity in shrimp comparing to lower protein diets. Thiobarbituric acid reactive substances (TBARS) of plasma were lower for shrimp fed on diets consisting of higher protein and energy levels than shrimp fed on the ₃₀P₁₉ diet. Analyses of digestive enzyme activities showed higher trypsin activities for shrimp fed on ₃₅P_17.5 and ₃₅P₁₉ diets comparing to ₃₀P₁₆ and ₃₀P_17.5 diets. Also, hepatopancreatic lipase activity of shrimp fed ₃₅P₁₆, ₃₅P_17.5 and ₃₅P₁₉ diets were significantly higher than those of shrimp fed the other diets. In conclusion, based on the results for growth performance, biochemical parameters, immune responses, oxidative stress and enzyme activities, 35% protein and 17.5 kJ/g digestible energy (₃₅P_17.5) could be considered as the optimum protein‐to‐energy ratio in the diet of juvenile whiteleg shrimp reared in a biofloc system.     

Mourilyan virus pathogenicity in kuruma shrimp (Penaeus japonicus)

The bunyavirus Mourilyan virus (MoV) occurs commonly in Black tiger (Penaeus monodon) and kuruma shrimp (Penaeus japonicus) farmed in eastern Australia. There is circumstantial evidence of MoV causing mortalities among P. japonicus moved from farm ponds to tanks for rearing as broodstock. To directly assess its pathogenic potential, independent cohorts of pond- (n = 24) or tank-reared juvenile (n = 21) P. japonicus were challenged intramuscularly with a cephalothorax tissue homogenate of P. monodon containing high loads of MoV (1.48 ± 0.28 × 10⁸ MoV RNA copies/µg total RNA). In each trial, mortalities accumulated gradually among the saline-injected controls. Mortality onset occurred 12–14 days earlier in the pond-reared shrimp, possibly due to them possessing low-level pre-existing MoV infections. Despite the time to onset of mortality differing, Kaplan–Meier survival analyses confirmed mortality rates to be significantly higher in both the pond- (p = .017) and tank-reared shrimp (p = .031) challenged with MoV. RT-qPCR data on shrimp sampled progressively over each trial showed high loads of MoV to establish following challenge and discounted GAV and other endemic viruses from contributing to mortality. Together, the data show that acute MoV infection can adversely compromise the survival of juvenile P. japonicus.     

Fishmeal levels can be successfully reduced in white shrimp (Litopenaeus vannamei) if supplemented with DL‐Methionine (DL‐Met) or DL‐Methionyl‐DL‐Methionine (Met‐Met)

The main objective of this study was to evaluate the effect of methionine supplementation when reducing fishmeal levels in diets for white shrimp (Litopenaeus vannamei). Tested diets consisted of a positive control with 260 g/kg fishmeal (D1), two negative controls with 100 g/kg fishmeal and no amino acid (AA) supplementation (D2) or supplemented with lysine but not methionine (D3), and four additional diets with 100g/kg fishmeal supplemented with increasing levels of DL‐Met (1.0, 2.0 or 3.0 g/kg) (D4, D5, D6) or Met‐Met (1.0 g/kg) (D7). Each diet was fed to four groups of 30 shrimp for 8 weeks at a daily rate of 70 g/kg body weight. Reduction in fishmeal from 260 g/kg down to 100 g/kg did not significantly affect survival rate, feed conversion ratio (FCR), protein efficiency ratio (PER) or protein retention efficiency (PR%) of white shrimp. However, growth performance (final body weight, FBW; weight gain, WG; specific growth rate, SGR) was reduced when dietary fishmeal level was reduced from 260 g/kg (D1) to 100 g/kg without methionine supplementation (D2). The growth performance (FBW, WG and SGR) of shrimp was significantly increased by supplementation of the 100 g/kg fishmeal diet with increasing levels of DL‐Met (p < .05). Same performance as positive control (D1) was achieved with diets containing 100 g/kg fishmeal and supplemented with 3.0 g/kg DL‐Met or 1.0 g/kg Met‐Met. The highest values of growth performance (FBW, WG and SGR) were found in shrimp fed D6 and D7 diets, which were significantly higher than those of shrimp fed D2 and D3 diets (p < .05) but without statistical differences with shrimp fed D1, D4 and D5 diets (p > .05). The highest values of whole‐body and muscle protein contents were found in shrimp fed D1 diet, which were significantly higher than those of shrimp fed all other diets (p < .05). The highest value of intestinal tract proteolytic enzyme activity was found in shrimp fed Met‐Met‐supplemented diet (D7) and followed by the positive control diet (D1) and 3 g/kg DL‐Met‐supplemented diet (D6) (p < .05). The highest values of apparent digestibility coefficients (ADCs) of dry matter and crude protein were found in Met‐Met‐supplemented diet (D7) and followed by the positive control diet (D1) (p < .05). Shrimp fed the D1 diet showed the highest value of total essential amino acid (EAA) and was significantly higher than shrimp fed D2–D3 (p < .05) but without significant difference with shrimp fed D4–D7 (p > .05). In conclusion, results showed that same performance can be achieved with diets containing 260 or 100 g/kg fishmeal supplemented with 3.0 g/kg DL‐Met or 1.0 g/kg Met‐Met. Moreover, supplementation of limiting methionine in low‐fishmeal diets seems to improve the digestive proteolytic activity, improving digestibility of dry matter and protein, and eventually to promote growth of juvenile white shrimp in fishmeal reduction diets.     

Dietary nucleotide‐rich yeast supplementation improves growth,innate immunity and intestinal morphology of Pacific white shrimp (Litopenaeus vannamei)

An 8‐week feeding trial was conducted to evaluate the effects of dietary nucleotide (NT)‐rich yeast supplementation on growth, innate immunity and intestinal morphology in Pacific white shrimp (Litopenaeus vannamei). Four isonitrogenous and isolipidic practical diets were formulated to contain 0 (control), 10, 30 and 50 g/kg of NT‐rich yeast, respectively. A total of 480 shrimp with an average initial body weight of 1.86 ± 0.02 g were randomly allocated into four groups, with four replicates per group and 30 shrimp each replicate. The results indicated that shrimp fed the diet containing 50 g/kg NT‐rich yeast had significantly higher weight gain (WG), specific growth rate (SGR) and protein efficiency ratio (PER) than those fed the control diet, and the lowest feed conversion ratio (FCR) was observed in the shrimp fed the 50 g/kg NT‐rich yeast supplemental diet. However, there was no significant difference in survival among all treatments. The crude protein of whole shrimp in the 50 g/kg NT‐rich yeast group was higher than that in the control group. Total protein, triglyceride concentrations, the activities of aspartate aminotransferase and alanine aminotransferase in serum were significantly influenced by the dietary NT‐rich yeast supplementation. The activities of serum phenoloxidase (PO) and lysozyme (LZM) of shrimp fed the diet containing 50 g/kg NT‐rich yeast were higher than those in shrimp fed the other diets. Relative expressions of alp and lzm significantly upregulated in the 30 g/kg NT‐rich yeast group compared to the control group. The intestinal fold height and fold width in the 30 g/kg NT‐rich yeast group were significantly higher than those fed the control diet; and the highest microvillus height occurred in the shrimp fed the 50 g/kg NT‐rich yeast diet. In summary, dietary 30–50 g/kg NT‐rich yeast supplementation promotes growth performance, enhances innate immunity and improves intestinal morphology of Litopenaeus vannamei.     

Effects of dietary astaxanthin on growth,antioxidant capacity and gene expression in Pacific white shrimp Litopenaeus vannamei

Pacific white shrimp Litopenaeus vannamei (1050 individuals with initial weight of 1.01 ± 0.001 g) were fed either control diet or one of six dietary astaxanthin (AX) concentration (25, 50, 75, 100, 125 and 150 mg kg⁻¹) diets for 56 days in 35 tanks (30 shrimp per tank). After 56 days of culture, shrimp‐fed AX125 and AX150 diets had higher (P < 0.05) weight gain, specific growth rate, total antioxidant status and lower (P < 0.05) superoxide dismutase (SOD), catalase (CAT) than shrimp fed control diet. After low dissolved oxygen stress for 1 h, survival rate of shrimp fed AX75, AX100, AX125 and AX150 diets was higher (P < 0.05) than that of shrimp fed control diet. Hypoxia inducible factor‐1α (HIF‐1α), cytosolic manganese superoxide dismutase (cMnSOD) and CAT mRNA expression levels of shrimp fed seven diets were significantly down‐regulated under hypoxia than under normoxia, but their expression levels were higher under hypoxia in shrimp fed AX‐supplemented diets than in shrimp fed control diet. About 70‐kDa heat‐shock protein (Hsp70) mRNA expression level of shrimp fed seven diets was significantly up‐regulated under hypoxia than under normoxia, but its expression level was lower under hypoxia in shrimp fed AX‐supplemented diets than in shrimp fed control diet.     

The value of sea urchin,Lytechinus variegatus,egesta consumed by shrimp,Litopenaeus vannamei

Sea urchins produce high‐energy, membrane‐bound fecal pellets that contain residual nutrients and large quantities of microbiota. These egesta are readily consumed by the shrimp, Litopenaeus vannamei. Egesta of the sea urchin, Lytechinus variegatus, were evaluated as a feed supplement or total replacement for a commercial shrimp diet. Shrimp were stocked at 0.49 g ± 0.06 g initial body weight and housed individually in 2.8‐L tanks in a commercial recirculating zebrafish system. Shrimp were assigned to one of six diets: commercial shrimp feed, reference sea urchin feed, collected dried sea urchin egesta, collected wet sea urchin egesta, half ration of shrimp feed and half collected wet sea urchin egesta, and egesta naturally produced by two sea urchins in polyculture. Equivalent dry matter amounts of each diet were proffered to shrimp in each treatment twice daily, except for those that had complete access to natural egesta excreted by sea urchins in polyculture. Sea urchins were proffered a reference sea urchin feed at 2% body weight daily. After 27 days, shrimp proffered collected dried or wet egesta did not differ significantly in percent weight gain and showed the lowest weight gain. The percent weight gain of shrimp fed the commercial shrimp diet did not differ significantly from that of the shrimp fed half commercial shrimp diet and half egesta. The highest weight gain was recorded for those shrimp that consumed the untouched egesta produced by sea urchins in polyculture. These data suggest that consumed egesta have noteworthy nutritional value and therefore would be beneficial to the culture of extractive species in an integrated multitrophic aquaculture system.     

Growth performance,haematological parameters,antioxidant status and salinity stress tolerance of juvenile Pacific white shrimp (Litopenaeus vannamei) fed different levels of dietary myo‐inositol

A 10‐week growth trial was run to evaluate effects of myo‐inositol (MI) on growth performance, haematological parameters, antioxidative capacity and salinity stress tolerance of Litopenaeus vannamei. Six practical diets supplemented with graded levels of MI (designated as MI0, MI600, MI1200, MI2400, MI 3600 and MI4800 for 448.8, 974.2, 1568.0, 2810.6, 3835.5 and 4893.6 mg/kg diet, respectively) were fed to six replicate groups of L. vannamei (mean initial body weight 0.63 ± 0.00 g). The results showed that significant increment of growth performance was observed in shrimp fed MI600 diet than those fed MI1200 diet. Lipid concentration in whole body of the shrimp fed MI600 diet was significantly increased. Shrimp fed MI0 diet had lower total protein (TP) as compared to shrimp fed the MI‐supplemented diets (except MI4800 diet). In general, lower activities of antioxidant enzymes and higher malondialdehyde (MDA) content in haemolymph and hepatopancreas were recorded in shrimp fed MI0 diet, compared to those fed the MI‐supplemented diets. Reduced survival after 7‐h salinity stress was present in shrimp fed MI0 diet as compared to those fed MI4800 diet. Dietary MI requirement for glutathione peroxidase activity of L. vannamei was 2705 mg/kg diet.     

Feed preference and growth response of juvenile Litopenaeus vannamei to supplementation of marine chemoattractants in a fishmeal‐challenged diet

This study compared the feed preference and growth response of Litopenaeus vannamei to chemoattractants. A diet with 3% fishmeal was supplemented with either 3% salmon meal (POS), 3% soy protein concentrate (NEG), 3% krill meal (KRM), 3% squid meal (SQM), 3% shrimp head meal (SHM), 3% shrimp meal (SM), 3% squid liver meal (SLM), or 5% liquid sardine hydrolysate (SAH). Shrimp with a body weight (BW) of 0.99 ± 0.08 g were stocked at 100 animals/m² in 56 tanks of 1 m³ and fed 10 times daily for 74 days. Feed preference was evaluated by feeding shrimp of 10.87 ± 1.82 g in excess twice a day for 10 days in two separate feeding trays allocated in 50 tanks of 0.5 m³. Survival reached 93.3 ± 5.80% and was unaffected by supplementation. Final BW was the highest for shrimp fed the KRM‐supplemented diet (11.97 ± 0.93 g), followed by POS (11.11 ± 0.77 g) and SQM (11.01 ± 1.17 g). Diets SHM, SM, SLM, and NEG showed a lower shrimp BW than POS, but were not statistically different among them. Shrimp fed the SAH diet achieved the lowest BW (10.06 ± 1.02 g). The highest gained yield was obtained with diets KRM and POS. No statistical difference was observed in shrimp yield among other diets. The lowest feed conversion ratio (FCR) was achieved with shrimp fed KRM (1.31 ± 0.05) when compared to diets SHM (1.47 ± 0.05), SAH (1.47 ± 0.07), and SLM (1.45 ± 0.17). Two‐by‐two comparisons indicated that shrimp preferred SHM and KRM, except when these were compared to SQM and SLM. No difference in feed preference was found between diets with SQM and SLM. SAH was the least preferred raw material in all comparisons. Results indicated that KRM acts as a powerful feeding effector and growth enhancer in fishmeal‐challenged diets for whiteleg shrimp. A dietary supplementation with 3% KRM is more effective than the same dose of any other chemoattractant evaluated.     

Effects of dietary β‐glucan and glycyrrhizin on non‐specific immunity and disease resistance of white shrimp,Litopenaeus vannamei (Boone) challenged with Vibrio alginolyticus

The white shrimp Litopenaeus vannamei, fed immunostimulant‐free, 0.2%β‐glucan and 0.06% glycyrrhizin diets for 18 days, respectively, were challenged with Vibrio alginolyticus at 6.4 × 10⁴ CFU shrimp^?1. The total haemocyte count (THC), phenoloxidase (PO) activity, respiratory burst (RB) and superoxide dismutase (SOD) activity changes for a 120‐h period were investigated, and shrimp mortality was also recorded. The results showed that PO activity, RB and SOD activity were significantly higher in shrimp fed the two immunostimulant diets after 18 days than those in shrimp fed immunostimulant‐free diets. The THC and SOD activity decreased significantly from 0 to 24 h post challenge, and then reverted to normal levels at 96 and 72 h respectively. The values for PO activity and RB increased from 0 to 48 h post challenge. Compared with those fed the control diets, shrimp fed immunostimulants had significantly higher PO activity and RB values at 120 h post challenge. Mortalities after challenge with V. alginolyticus were significantly lower in shrimp fed with β‐glucan or glycyrrhizin than in those fed with a diet without immunostimulants. It was concluded that dietary β‐glucan and glycyrrhizin increased the shrimp immunity. Furthermore, β‐glucan caused an increase in some immune parameters 12 h earlier than glycyrrhizin after V. alginolyticus challenge.     

Enhanced growth effects on shrimp (Litopenaeus vannamei) from inclusion of whole shrimp floc or floc fractions to a formulated diet

A growth trial was conducted to determine the effects of inclusion of whole shrimp floc or floc fractions to a control diet on growth and survival of shrimp (Litopenaeus vannamei). The floc sample was collected from marine shrimp culture tanks and partially fractionated by extraction with water, acetone and hexane. A series of diets was manufactured by inclusion of whole floc (intact or ground), each of the fractions or their combination to a control diet. These diets were fed to shrimp (approximately 1.0 g) in an indoor laboratory under flow‐through conditions for 8 weeks. It was found that addition of whole floc (200 g kg^?1) or floc fractions (24–200 g kg^?1) to the control diet improved (P < 0.05) shrimp growth rate without affecting (P > 0.05) shrimp survival (>81.3%). Although inclusion of whole floc reduced the crude protein and crude fat contents and gross energy of the control diet, shrimp fed the whole floc‐supplemented diets obtained the highest (P < 0.05) growth rates (1.01 and 1.03 g week^?1) among the shrimp fed the 11 tested diets including two control (0.81 and 0.85 g week^?1), two commercial (0.45 and 0.71 g week^?1) and five floc‐fraction‐added (0.91–1.00 g week^?1) diets. Many bioactive compounds in the floc that possibly affected shrimp growth were also analysed and quantified.     

Optimal dietary lipid requirement of postlarval white shrimp,Litopenaeus vannamei in relation to growth performance,stress tolerance and immune response

A 25‐day experiment was conducted to evaluate the optimal lipid level for postlarval Litopenaeus vannamei. Shrimp (1.7 mg) were fed five isonitrogenous diets containing grade levels of lipid (96.6, 114.3, 128.5, 136.5 and 154.5 g/kg diet, respectively). Each diet was assigned to four tanks (500 shrimp), and shrimp were fed six times a day. Weight gain was increased with the increasing dietary lipid levels, and the highest weight gain was observed in shrimp fed diet with 154.5 g/kg lipid (p < 0.05). On the contrary, the survival was lowest in shrimp fed the L15.45 and highest in shrimp fed the L11.43. Triglyceride in hepatopancreas was increased, and cholesterol was decreased with the increasing dietary lipid. Pyruvate kinase and AMPK mRNA expression were highest in shrimp fed the L12.85. Malondialdehyde in whole body was positively correlated with the dietary lipid levels. The mRNA expression of SOD and Caspase 3 was highest in shrimp fed the L12.85. After hypoxia stress, shrimp fed the L12.85 showed highest survival. The mRNA expression of superoxide dismutase and Akirin was highest in shrimp fed the L11.43 and L15.45, respectively. Based on the survival after 25‐day feeding trail and after the hypoxia stress, the optimal dietary lipid for postlarval L. vannamei should be 118–124 g/kg.     

Anti‐PirA‐like toxin immunoglobulin (IgY) in feeds passively immunizes shrimp against acute hepatopancreatic necrosis disease

Acute hepatopancreatic necrosis disease (AHPND), caused by a toxin‐producing Vibrio parahaemolyticus strain, has become a serious threat to shrimp aquaculture. The need to regulate antibiotic use prompted the development of alternative ways to treat infections in aquaculture including the use of chicken egg yolk immunoglobulin (IgY) for passive immunization. This study evaluated the protective effect of IgY against AHPND infection in Litopenaeus vannamei (Boone). IgY was isolated from eggs laid by hens immunized with recombinant PirA‐like (rPirA) and PirB‐like (rPirB) toxins. Whole‐egg powders having IgY specific to rPirA (anti‐PirA‐IgY) and rPirB (anti‐PirB‐IgY) and IgY from non‐immunized hen (control‐IgY) were mixed with basal diets at 20% concentrations and used to prefeed shrimp 3 days before the bacterial challenge test. Survival rates of the challenged shrimp fed the anti‐PirA‐IgY, anti‐PirB‐IgY and control‐IgY diets were 86%, 14% and 0%, respectively. Only the feed containing anti‐PirA‐IgY protected shrimp against AHPND. Increasing the concentration of rPirA antigen to immunize hens and lowering the amount of egg powder in feeds to 10% consistently showed higher survival rates in shrimp fed with anti‐PirA‐IgY (87%) compared with the control (12%). These results confirm that addition of anti‐PirA‐IgY in feeds could be an effective prophylactic method against AHPND infection in shrimp.     

MATURATION,REPRODUCTION, AND GROWTH OF Penaeus vannamei AND P. stylirostris FED NATURAL DIETS

A variety of natural foods have been reported to induce ovarian maturation of penaeid shrimp. In this study, conducted from 19 June to 11 September 1980, four single-food diets (clams, shrimp, squid, and worms) and one composite diet consisting of all four foods were fed to Penaeus vannamei in 3.7-m diameter indoor circular tanks. The shrimp, averaging 28 g, were stocked at a rate of 6.9 m^-2 with a 1:1 male to female ratio. Unilateral eyestalk ablation was performed on half of the females in each tank. Tanks received one turnover of water volume daily and the water temperature and salinity averaged 27.0°C and 28.6 ppt, respectively. Daily photoperiod was maintained at 13 hours of light. Ovarian maturation, spawning, molting rate, and survival were monitored daily and growth and gonad size were measured at termination. The composite diet was the best overall diet, while squid was the best single-food diet, followed by shrimp, worms, and clams. No maturation of unablated females was observed with the exception of one female that developed to stage 3. Although the size of the shrimp was considered marginal for spawning, 5 mated females were observed, but only one (squid treatment) produced viable nauplii. After termination of this study, all shrimp were fed the composite diet for 41 days. During this period, ovarian maturation and spawning occurred in the previously unproductive treatment receiving only clams, and growth rate of shrimp in all previously single-food treatments increased 4 to 7 fold. The composite diet was also fed to two tanks of P. stylirostris (mean initial weight = 45.4 g) for a period of 118 days. During that time, 50 spawns were obtained with up to 3 spawns per female. A total of 4.3 × 10⁵ P. stylirostris nauplii were produced.     

Replacement of Fish Meal with Soybean Meal,Alone or in Combination with Distiller's Dried Grains with Solubles in Practical Diets for Pacific White Shrimp,Litopenaeus vannamei,Grown in a Clear‐Water Culture System

The objective of this study was to evaluate inclusion of distiller's dried grains with solubles (DDGS) as partial replacement of commercial, solvent‐extracted soybean meal (SBM) in fish meal‐free diets for Pacific white shrimp, Litopenaeus vannamei. Aquaria connected to a recirculating biofiltration system were utilized to evaluate growth, survival, and feed conversion of shrimp during the 8‐wk feeding trial. Each 110‐L aquarium was stocked with 15 shrimp (mean individual weight 0.99 g) and fed one of five diets: a diet containing 20% fish meal (FM), which served as the control (Diet 1); a diet containing 0% FM and 52.5% SBM (Diet 2); and diets containing 0% FM and either 10, 20, or 30% DDGS as partial replacement of SBM (Diets 3, 4, and 5, respectively). Shrimp were fed according to a pre‐determined feeding chart five times daily (0730, 1030, 1330, 1630, and 1930 h) and there were three replicates per dietary treatment. The results from the feeding trial demonstrated that final weight, weight gain (g), and percentage weight gain were significantly higher (P < 0.05) for shrimp fed Diet 1 (10.96 g, 10.01 g, and 1051%, respectively) compared to shrimp fed diets containing DDGS; however, shrimp fed diets containing DDGS had similar (P > 0.05) final weight, weight gain (g), and percentage weight gain as shrimp fed a diet containing 0% FM and 52.5% SBM (Diet 2). Feed conversion ratio (FCR) of shrimp fed Diet 1 (2.84) was significantly lower (P < 0.05) compared to shrimp fed any other diet. Survival (%) was not different (P > 0.05) among treatments and averaged 77.3% for the study. This study demonstrated that practical shrimp diets containing no FM had an adverse impact on growth performance of white shrimp when grown in a clear‐water system and that further research is needed to refine diet formulations when culturing shrimp in these systems when attempting to feed a diet without FM.     

Effects of dietary administration of Macsumsuk® on growth and stress to low salinity and low dissolved oxygen in whiteleg shrimp,Litopenaeus vannamei (Boone, 1931) juveniles

An 8‐week study was conducted to explore the results of Macsumsuk^® as a feed additive on the stress tolerance and growth of Litopenaeus vannamei in 15 culture tanks of 36 L each. Three hundred shrimp averaging 0.1 ± 0.01 g were fed with five isonitrogenous (48.38 ± 0.38% CP) diets (in triplicate groups) containing kaolinite (Macsumsuk^®) at 0%, 0.3%, 0.6%, 1.2% and 2.4%, namely Mk₀, Mk_0.3, Mk_0.6, Mk_1.2 and Mk_2.4. Specific growth rate (SGR) and weight gain (WG) of shrimp fed diets Mk_1.2 and Mk_2.4 were significantly better than those of shrimp fed diet Mk₀ (p < .05). However, SGR and WG of shrimp fed diets Mk_0.6, Mk_1.2 and Mk_2.4 were not significantly different. Protein efficiency ratio (PER) and feed efficiency (FE) of shrimp fed diets Mk_1.2 and Mk_2.4 were significantly better than those of shrimp fed diets Mk₀, Mk₀.₃ and Mk_0.6. Furthermore, the survival of shrimp fed diet Mk_2.4 was significantly lower than that of shrimp fed diet Mk_0.6 (p < .05). Cumulative mortality of shrimp fed diet Mk_1.2 was significantly lower than that of shrimp fed diet Mk₀ at 1–1.5 hr post‐stress to low dissolved oxygen (from 6.1 mg/L to 2.9 mg/L) and 4–5 hr post‐stress to low salinity (from 32‰ to 1‰) (p < .05). The optimum dietary Macsumsuk^® level for juvenile L. vannamei was determined as 1.97% by the polynomial regression analysis of weight gain.     

Effects of dietary dl‐methionyl‐dl‐methionine (Met‐Met) on growth performance,body composition and haematological parameters of white shrimp (Litopenaeus vannamei) fed with plant protein–based diets

A 10‐week feeding trial was conducted to evaluate the effects of supplementing different levels of dl ‐methionyl‐dl ‐methionine (AQUAVI^® Met‐Met) in plant protein–based diets on Litopenaeus vannamei. The positive control (PC) and negative control (NC) diets were designed with 20% and 8% fishmeal respectively, and other six diets were formulated with graded levels of Met‐Met from 0.05% to 0.30% with a 0.05% increment on the basis of NC diet (MM 0.05–MM 0.3). Six replicates were randomly assigned to each diet with 50 shrimp each having initial weight of 0.98 ± 0.02 g. The variation of FM concentration from 20% to 8% and supplemented with graded levels of Met‐Met did not affect the survival rate, feed conversion ratio, protein efficiency ratio, whole body and muscle proximate compositions (p > 0.05). However, diets with ≤0.20% Met‐Met supplementation resulted in significantly increased weight gain and specific growth rate, after which both parameters reached plateau. Shrimp fed the NC diet showed significantly lower total essential amino acid (EAA) content in muscle (p < 0.05). Supplementation of Met‐Met significantly improved apparent digestibility coefficients of dry matter, crude protein, lipid, phosphorus and EAAs (p < 0.05). Based on broken‐line analysis, the methionine requirement for white shrimp was estimated to be 0.87% when using Met‐Met as methionine source.     

Comparison of L‐lysine·HCl and L‐lysine sulphate in the feed of Penaeus monodon and re‐evaluation of dietary lysine requirement for P. monodon

Two trials were conducted to compare L‐lysine HCl and L‐lysine sulphate regarding its availability to Penaeus monodon, and further evaluate the optimum dietary lysine requirement. In experiment 1, five experimental diets were formulated (D1, D2, D3, D4 and D5), a basal diet (D1), aimed at a low‐lysine concentration (2.22% dry matter), with lysine concentration of the other four diets increasing in two 0.25% L‐lysine intervals from either L‐lysine HCl (D2 and D3) or L‐lysine sulphate (D4 and D5). Each diet was fed at a restricted rate to three groups of 40 shrimp for 74 days. The highest values of growth performance (weight gain, WG; specific growth rate, SGR) and survival were observed with shrimp fed the L‐lysine HCl diet. Feed efficiency (FE) of shrimp fed D2 was significantly higher than that of shrimp fed D1 and D5 (P < 0.05), but without significant difference with shrimp fed D3 and D4 (P > 0.05). In experiment 2, six diets (d1, d2, d3, d4, d5 and d6) were formulated with six graded levels of lysine (2.21%, 2.41%, 2.59%, 2.87%, 3.11% and 3.29% of diet). Each diet was randomly assigned to triplicate groups of 40 shrimp for 74 days. WG, SGR and survival increased increasing levels of lysine up to 2.41% of diet and reached an apparent plateau. Broken‐line model analysis on WG and SGR indicated that the optimum dietary lysine level for optimal growth of shrimp was 2.37% of diet, corresponding to 5.78% of dietary protein. In conclusion, results of this trial suggest that L‐lysine HCl is superior to L‐lysine sulphate when fed to Penaeus monodon and optimal growth can be obtained at lysine levels corresponding to 2.37% of diet, or 5.78% of dietary protein in this specie.     

Oral Administration of Hot‐water Extract of Tropical Brown Seaweed,Sargassum cristaefolium,to Enhance Immune Response,Stress Tolerance,and Resistance of White Shrimp,Litopenaeus vannamei,to Vibrio parahaemolyticus

The efficacy of hot‐water extract of tropical brown seaweed, Sargassum cristaefolium (SCE), supplemented in diets on immune response, stress tolerance, and disease resistance of Litopenaeus vannamei to Vibrio parahaemolyticus was evaluated. Shrimp were fed diets containing graded levels of SCE (0, 250, 500, 750, and 1000 mg/kg). The results showed that shrimp fed all diets containing SCE had significantly higher (P < 0.05) immune response in total hemocyte count (THC), differential hemocyte count (granular and hyaline cells), and phagocytic activity than those of shrimp fed the control diet. Similarly, in low dissolved oxygen stress tolerance test and the challenge test with V. parahaemolyticus, survival rates of shrimp fed all diets containing SCE were significantly higher (P < 0.05) (83–93% in stress test and 27–47% in challenge test) than those of shrimp fed the control diet (77 and 3.3%, respectively). These results suggest that oral administration of SCE at 500 and 750 mg/kg can be effectively used to enhance immune response, stress tolerance, and resistance of white shrimp, L. vannamei, against V. parahaemolyticus infection. These findings also confirm that using dietary SCE as immunostimulant is effective at increasing the nonspecific immune system in penaeid shrimp, L. vannamei.     

Suitable dietary chitosan improves the growth performance,survival and immune function of tiger shrimp,Penaeus monodon

Different levels of dietary chitosan on growth performance, survival and stress tolerance to air exposure was studied in tiger shrimp, Penaeus monodon. Shrimp (mean initial wet weight about 1.16 g) were fed with six different diets (C0, C0.05, C0.1, C0.2, C0.3 and C0.4) containing six level of chitosan (0%, 0.05%, 0.1%, 0.2%, 0.3% and 0.4% respectively) in triplicate for 60 days. Growth performance [final body wet weight (FBW); weight gain (WG); biomass gain (BG)] of shrimp fed chitosan‐containing diets were higher (P < 0.05) than that of shrimp fed the basal diet, shrimp fed C0.1 diet showed the highest value of growth performance. Survival of shrimp in C0.1 and C0.2 diet groups were higher (P < 0.05) than that of shrimp in C0, C0.05 and C0.4 diet groups but without statistical difference (P > 0.05) in shrimp fed C0.3 diet group. Whole body and muscle lipid contents decreased with increasing dietary chitosan levels. Plasma total cholesterol and triglyceride contents of shrimp fed C0 diet was significantly higher (P < 0.05) than that of shrimp fed chitosan‐containing diets. Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities of shrimp fed C0 diet were higher than those of shrimp fed chitosan‐containing diets. Digestive gland malondialdehyde (MDA) and carbonyl protein contents of shrimp fed chitosan‐containing diets were lower (P < 0.05) than that of shrimp fed C0 diet. Total haemocyte count of shrimp fed C0 diet was lower (P < 0.05) than that of shrimp fed chitosan‐containing diets. On the contrary, the haemolymph clotting time of shrimp fed C0 diet was higher (P < 0.05) than that of shrimp fed chitosan‐containing diets. In conclusion, all results suggested that dietary intake containing 0.1% and 0.2% chitosan enhanced the growth of shrimp, whereas a higher level than 0.3% and 0.4% decreased growth of shrimp. Second‐degree polynomial regression analysis of WG and BG indicated that the optimum supplement of dietary chitosan level should be 0.19–0.21%.