Response of Pacific white leg shrimp (Litopenaeus vannamei) on exposure to aflatoxin in feed

A research was accomplished to find the response of Litopenaeus vannamei to different levels of aflatoxin in feed. Two hundred live L. vannamei were collected from a shrimp breeding station located on the south of Tiab, in Hormozgan province. Aflatoxin contaminated diets with 0.18, 96.21, 184.74, 711.35, 977.11 and 1605.61 µg/kg were prepared and shrimps in six experimental groups with three replicates in each group for 28 days were fed with these diets. The results showed that activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) in haemolymph and histological alternation index (HAI) value of hepatopancreas in treatement groups were significantly higher than control group (p < 0.05). Decreased total protein and fat in haemolymph and increased protein in muscle of treatment shrimps were observed compared with the control group (p < 0.05). The HAI value of 977.11 and 1605.61 μg/kg groups were significantly higher than the control and other groups (p < 0.05). Histological study results showed that calculated HAI (more than 100) in these groups as a result of necrosis lesion observed in hepatopancreas. Moderate to severe alteration was recognized in group fed by 711.35 µg/kg aflatoxin. Minor aflatoxin levels (0.18, 96.21 and 184.74 µg/kg) caused inappreciable tissue alteration. This study demonstrated body composition, haemolymph parameters and histology of hepatopancreas were affected in the aflatoxin‐contaminated feed when the concentration was over 955.14 µg/kg.     

Effects of dietary nucleotides on growth,survival and metabolic response in whiteleg shrimp,Litopenaeus vannamei against ammonia stress condition

This trial was conducted to evaluate the effects of nucleotides on growth of whiteleg shrimp, Litopenaeus vannamei, and the survival and metabolic responses to ammonia stress test. Experimental diets were as follows: low fish meal diet (LFMD), and four LFMD test diets, each supplemented with 0.1% guanosine monophosphate (GMP), 0.1% inosine monophosphate (IMP), 0.1% mixture of GMP and IMP and 0.1% mixture of GMP, IMP, uridine monophosphate (UMP) and cytidine monophosphate (CMP). The shrimp specimens (initial body weight: 0.99 ± 0.01 g) were randomly allocated into five groups and fed four times daily for 8‐weeks. After the trial, final body weight was recorded and haemolymph was withdrawn for haematological analysis. The shrimp was then challenged with 70 mg/L ammonia (LC50) for 10 days. Survival and haemolymph of the shrimp were taken after exposure to ammonia. The highest growth performance was observed in the shrimp fed diet supplemented with GMP (p < .05), while survival was not influenced by the test diets in the feeding trial. In the ammonia challenge test, the highest survival was observed in the shrimp fed GMP supplemented diet compared to others. The plasma protein, glucose and cholesterol levels increased in all the treatments while triglycerides level decreased post challenge. Cortisol level recovered at day 10th after the challenge. Shrimps fed with nucleotides diets showed higher protein and glucose level compared to control groups post challenge. In general, nucleotides supplemented in the diet enhanced growth, improved stress resistance while modulating the haemolymph metabolites in L. vannamei under ammonia stress.     

Antioxidant effects of nanoencapsulated lipoic acid in tissues and on the immune condition in haemolymph of Pacific white shrimp Litopenaeus vannamei (Boone, 1931)

The antioxidant and oxidative damage responses and immune parameters in shrimp, Litopenaeus vannamei, reared in clear water (CW) and in a biofloc system (BFT) were evaluated. The animals were given feed supplemented with either nanocapsules containing the antioxidant lipoic acid (NCLA) or empty nanocapsules (NC). Antioxidant and oxidative stress analyses were evaluated by measuring glutathione S‐transferase activity (GST), reduced glutathione (GSH) concentration and lipid peroxidation (TBARS) in the gills, hepatopancreas and muscle. The immune parameters were then determined by the differential hemocyte count in the haemolymph. The final weight of the shrimp reared in both CW and BFT and treated with NCLA increased (p < .05). The shrimps reared in BFT showed increased GST activity in the gills and hepatopancreas after NCLA treatment (p < .05). Higher GSH levels were observed in the hepatopancreas with NCLA supplementation in CW and BFT (p < .05). Decreased levels of TBARS were observed in the gills and muscles of shrimp supplemented with NCLA and maintained in CW (p < .05). A decrease in the percentage of hyaline hemocytes and an increase in granular hemocytes were observed in shrimp treated with NCLA in CW and BFT (p < .05). The antioxidant treatment induced positive responses in terms of the zootechnical, biochemical and immunological parameters.     

Replacement of fish oil with a DHA‐rich Schizochytrium meal on growth performance,activities of digestive enzyme and fatty acid profile of Pacific white shrimp (Litopenaeus vannamei) larvae

This trial was conducted to evaluate the effects of replacing dietary fish oil with Schizochytrium meal for Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Six test microdiets were formulated using Schizochytrium meal to replace 0 g/kg, 250 g/kg, 500 g/kg, 750 g/kg, 1000 g/kg or 1500 g/kg fish oil DHA. No significant differences were observed in survival, growth, final body length and activities of digestive enzyme among shrimp fed different diets (p > .05). No significant differences were observed in C20:5n‐3 (EPA) in muscle samples (p > .05). C18:3n‐3 and C20:4n‐6 in muscle increased as Schizochytrium meal replacement level increased (p < .05). No significant differences were observed in C22:6n‐3 (DHA) and n‐3 fatty acids among shrimp fed diets that algae meal replaced 0 g/kg ‐ 1000 g/kg of fish oil. Shrimp fed diet R150 had higher DHA content than other groups and had higher n‐3 fatty acids than that of shrimp fed diets R50, R75 and R100 (p < .05). C18:2n‐6, PUFA and n‐6 fatty acids in muscle increased, while n‐3/n‐6 ratio decreased with increasing algae meal replacement level from 0 g/kg to 1000 g/kg (p < .05). In conclusion, Schizochytrium meal could replace 1500 g/kg fish oil DHA in the microdiets without negatively affecting shrimp larvae survival, growth and activities of digestive enzyme.     

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.     

Effects of dietary supplementation of Astragalus membranaceus,Codonopsis pilosula,and Glycyrrhiza uralensis extract mixture on growth performance,haematological parameters and hepatopancreatic performance in juvenile Pacific white shrimp (Litopenaeus vannamei)

Pacific white shrimp is the major farmed shrimp species in the world. It is known to be very sensitive to the environmental and management changes such as intensification, which is one of the primary necessities to increase shrimp production, but represents a stressful condition, and needs to be managed properly to diminish its negative effects in aquaculture. In this study, juvenile Pacific white shrimp were fed diets supplementing with equal quantities of Astragalus membranaceus, Codonopsis pilosula and Glycyrrhiza uralensis extracts in different concentrations (0, 1.0, 2.0 and 3.0 g/kg) for 8 weeks. We subsequently estimated the effects of supplemented diets on growth performance, haematological parameters, and histological changes of hepatopancreas. Results revealed that the supplemented mixture had no benefits on growth and survival rate, but had favourable effects on haemocyte count, number of granulocytes, and hemocyanin concentration in the haemolymph (p > 0.05). We observed that R cells and B cells were increased in hepatopancreatic tubules of shrimps fed on supplemented diets. Besides, a diet containing 2.0 g/kg of extract mixture maintained the decreased levels of alanine aminotransferase (p < 0.05) and aspartate aminotransferase (p > 0.05). Among the used concentrations, 2.0 g/kg seemed to be the most suitable concentration.     

Effect of lactic acid on enrofloxacin pharmacokinetics in Eriocheir sinensis (Chinese mitten crab)

As a representative acidifier, lactic acid (LA) is widely used in the diets of aquatic animals. LA is the main supporter of energy metabolism and may be associated with drug metabolism. This study was designed to explore the pharmacokinetics of enrofloxacin (ENR) in Eriocheir sinensis (Chinese mitten crab) fed diets containing 0.1%, 0.2% and 0.3% LA (designated groups LA1, LA2 and LA3 respectively). The concentrations of ENR in the haemolymph, hepatopancreas and muscle were determined by HPLC after a single oral dose of 10 mg/kg. Our results showed that LA had a significant effect on the peak ENR concentrations in all the tissues (p < 0.05) by one‐way ANOVA analysis. There was a trend that C_max (peak concentration) of ENR was elevated with LA levels increasing up to 0.3% in haemolymph, hepatopancreas, muscle and T_max (time‐point of the peak concentration of the drug), t_1/2β (elimination half‐life) and AUC_(0‐∞) of ENR were shortened. Taken together, 0.3% LA might be effective in improving ENR pharmacokinetics in E. sinensis. Furthermore, it can be speculated that the enhanced biotransformation of ENR in the hepatopancreas mediated by LA is responsible for the differences in the pharmacokinetics of ENR in E. sinensis.     

Aspergillus niger fermented plant protein mix as a potential substitute for fishmeal in the diet of Penaeus vannamei (Boone, 1931)

A 45‐day feeding trial was conducted to evaluate the utilization of fermented plant protein mix (FPPM) as a fishmeal substitute in the diet of Penaeus vannamei. Six isonitrogenous and isolipidic diets were formulated by substituting fishmeal at 0% (control), 50%, 60%, 70%, 80% and 90% (FR‐0, FR‐50, FR‐60, FR‐70, FR‐80 and FR‐90, respectively) using FPPM. Shrimps fed with diets FR‐50, FR‐60 and FR‐70 had no significant difference in weight gain (173.22%–178.07%) with the control group (179.50%). However, the broken‐line regression indicated that 70.8% fishmeal could be substituted by FPPM. The trend of digestibility was reflected in growth performance. Survival had no significant difference among the treatments. Fishmeal substitution had significant (p < .05) variations in protease and amylase activity, but not in lipase. Except lipid, other body composition was not affected due to dietary change. A decrease (p < .05) in haemolymph total protein, cholesterol and triglycerides was observed to increase in fishmeal substitution. A higher fishmeal substitution (FR‐90) had significantly (p < .05) lower total haemocyte count and the reverse was true for phenoloxidase. Results conclude that a combination of fermented ingredients would be more effective rather using alone in shrimp feed, with a great potential in reducing the pressure on fishmeal.     

Effects of freeze‐dried powder of the Antarctic krill Euphausia superba on the growth performance,molting and fatty acid composition of the Pacific white shrimp Litopenaeus vannamei

This study was designed to evaluate the effect of dietary replacement fish meal supplemented with freeze‐dried powder of the Antarctic krill Euphausia superba (FDPE) on the growth performance, molting, and fatty acid composition of the Pacific white shrimp Litopenaeus vannamei (initial weight 1.27 ± 0.09 g). Four diets containing 0% (S0 group), 10% (S10 group), 20% (S20 group), and 30% (S20 group) FDPE were used in the present study. At the end of growth trial, the final body weight, weight gain rate, and specific growth rate in the S10, S20, and S30 groups were higher than those in the S0 group. The shrimp in the S10 and S20 groups exhibited better molting synchronism than those in the S0 group. The astaxanthin content in the hepatopancreas from the shrimp in the groups supplemented with FDPE was significantly higher than that in the S0 group (p < 0.05) and increased as the FDPE content in the feed increased. The shrimp in the S10, S20, and S30 groups had a higher monounsaturated fatty acid (MUFA) content in the hepatopancreas than those in the S0 group. The sum of EPA and DHA in the muscles from the shrimp in the S0 group was lower than that in the other groups. These results indicate that the dietary inclusion of 10%–20% FDPE can be used as practical diets in L. vannamei farmed under a clear water system.     

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%.     

Supplementation of dietary apple cider vinegar as an organic acidifier on the growth performance,digestive enzymes and mucosal immunity of green terror (Andinoacara rivulatus)

The present study was designed to investigate the effects of dietary apple cider vinegar (ACV) on digestive enzyme activity and growth performance as well as immune responses and antibacterial activity of skin mucus in green terror (Andinoacara rivulatus). Fish were fed diets supplemented with 0%, 1%, 2% and 4% of ACV (40.830 ppm acetic acid concentration) for 63 days. The final weight and weight gain values were observed to be significantly higher in fish fed with 2% of ACV compared to the control group (p < .05). ACV inclusion in the diets had significant effects on SGR (%) and FCR values (p > .05). ACV treatment resulted in a significant increase in the intestinal protease, α‐amylase, lipase and alkaline phosphatase activities compared to control (p < .05). The activities of digestive enzymes in fish fed with 2% and 4% of ACV diets were significantly higher than the other groups (p < .05). The total protein content, alternative haemolytic complement, alkaline phosphatase, total immunoglobulins and lysozyme activities of skin mucus increased significantly in fish fed with ACV diets (p < .05). In conclusion, administration of ACV enhanced digestive enzyme activity, growth performance, immune responses and the immune properties of skin mucus, and it can be used as a natural growth promoter and immunostimulant in green terror culture.     

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 herbal formulae combined by Astragalus polysaccharides,chlorogenic acid and allicin in different combinations and proportions on growth performance,non‐specific immunity,antioxidant status,vibriosis resistance and damage indexes of Litopenaeus vannamei

In this study, a feeding trial followed by a challenge test was performed to evaluate effects of six herbal formulae which were different combinations of Astragalus polysaccharides (APS), chlorogenic acid (CGA) and allicin on growth performance, non‐specific immune response, antioxidant capacity, disease resistance and biomolecule damage of white shrimp Litopenaeus vannamei. Shrimp were fed seven diets, control diet (basal diet with no herbal formulae) and six herbal formula diets (G1–G6, basal diet supplemented with 0.1% APS + 0.05% allicin, 0.1% APS + 0.1% allicin, 0.1% CGA + 0.05% allicin, 0.1% CGA + 0. 1% allicin, 0.1% APS + 0.1% CGA and 0.1% APS + 0.1% CGA + 0.05% allicin respectively), for 21 days. After that, shrimp were challenged with Vibrio harveyi and then the cumulative mortality of shrimp was recorded for 7 days post challenge. The results showed that there were no significant differences in growth performance among all groups, while the non‐specific immune responses and antioxidant indexes were significantly improved (p < .05) in shrimp fed herbal formula diets when compared to the control. Meanwhile, the lowest cumulative mortality was observed in shrimp fed herbal formula diets with 0.1% APS, 0.1% CGA and 0.05% allicin supplementation after V. harveyi challenge. Additionally, herbal formulae could not cause biomolecule damage to the hepatopancreas of shrimp. In conclusion, these results indicated that synergistic effect of APS, CGA and allicin helped to boost immunity, antioxidant capacity and disease resistance of shrimp without biomolecule damage.     

Developing a low fishmeal diet for juvenile Pacific white shrimp,Litopenaeus vannamei,using the nutritional value of FM as the reference profile

An 8‐week feeding trial was conducted to evaluate the effect of fish‐meal replacement on growth performance, antioxidative ability, immune capacity and haemolymph metabolites of Litopenaeus vannamei. A 250 g/kg fish‐meal diet was used as control (Diet 1). Other two diets contained 100 g/kg fishmeal, Diet 2 was supplemented with three indispensable amino acids, and Diet 3 was supplemented with microbalance components and phytase. Weight gain was lower in shrimp fed the Diet 2 than shrimp fed the Diet 1. Feed efficiency, protein efficiency ratio and survival were higher in shrimp fed the Diet 1 than other groups. Glutathione and nitric oxide contents in haemolymph were decreased in shrimp fed the Diet 2, and nitric oxide synthetase, phenoloxidase, acid phosphatase and myeloperoxidase activities showed the similar tendency. In hepatopancreas, antioxidative ability and immune capacity also lower in shrimp fed the Diet 2. In haemolymph, 21 differential metabolites were identified, and 13 metabolism pathways were affected by different diets. Generally, the fishmeal inclusion level could be reduced from 250 g/kg to 100 g/kg without affecting the growth performance of L. vannamei, and meanwhile, the protein utilization efficiency and antioxidative ability of shrimp fed the low fishmeal diets should be more focused on in future studies.     

Effects of different levels of dietary protein on the growth performance,nutrient utilization,digestive enzymes and physiological status of white shrimp,Litopenaeus vannamei juveniles reared in inland saline water

A 60‐day feeding trial was conducted to illustrate the effect of dietary protein levels on the growth and physio‐metabolic responses of juvenile Litopenaeus vannamei reared in inland saline water (ISW). Six isoenergetic (15 MJ/kg) and isolipidic (60 g/kg) diets with graded level of crude protein viz., 240 (T240), 260 (T260), 280 (T280), 300 (T300), 320 (T320) and 340 (T340) g/kg diet were formulated. Significantly higher (p < .05) weight gain (%), specific growth rate, with lower food conversion ratio were found in T320 and T340 groups. The protein utilizing efficiency and whole‐body protein content were significantly higher (p < .05) in the T320 group. Trypsin activity increased with the increasing dietary CP level but amylase activity decreased with the increasing dietary CP level. Transaminase enzymes, haemolymph protein and haemocyanin were elevated in T320 and T340 groups. The lactate dehydrogenase and malate dehydrogenase activities were significantly higher (p < .05) in the T240 group. Shrimp of T240 group had the lowest (p < .05) glycogen and total haemocyte count with highest (p < .05) haemolymph glucose and antioxidant enzymes activities than the other groups. Based on the results, feeding 320 g CP/kg is found to be optimum for supporting maximum growth and health status of L. vannamei reared in ISW at 8 g/L salinity. The finding of the present study will help in developing a low‐cost feed for L. vannamei reared in ISW.     

Improvement of growth performance,water quality and disease resistance against Vibrio harveyi of postlarval whiteleg shrimp (Litopenaeus vannamei) by administration of mixed microencapsulated Bacillus probiotics

The objective of this study was to investigate the effects of mixed Bacillus on growth, water quality and disease resistance against Vibrio harveyi in whiteleg shrimp (Litopenaeus vannamei). Postlarval shrimp (PL₃₀) were fed with (a) a basal diet (the control), (b) a diet containing mixed freeze‐dried Bacillus probiotics (FB) and (c) addition of mixed microencapsulated Bacillus probiotics (MB) in culture water. Addition of FB and MB probiotics improved (p < .05) growth, feed efficiency, survival and culture water quality (ammonia and nitrite) compared to the control group although there was no difference (p > .05) between the two treated groups. Bacillus numbers in gastrointestinal tracts and culture water of FB‐ and MB‐administrated shrimp were higher (p < .05) than in the control. After a 30‐day culture, shrimp were infected with V. harveyi and monitored for 10 days. A significant reduction (p < .05) in cumulative mortality was observed in FB‐ and MB‐supplemented shrimp (43.24% and 45.05%, respectively), compared to the control (63.06%). This finding demonstrated that administration of microencapsulated probiotics was as effective as freeze‐dried probiotics for improving growth, feed efficiency, survival, Bacillus in gastrointestinal tracts, water quality (ammonia and nitrite) and conferring disease resistance to V. harveyi.