Effects of dietary chlorogenic acid on growth performance, antioxidant capacity of white shrimp Litopenaeus vannamei under normal condition and combined stress of low-salinity and nitrite.

An eight-week feeding trial followed by an acute combined stress test of low-salinity and nitrite were performed to evaluate effects of chlorogenic acid (CGA) on growth performance and antioxidant capacity of white shrimp Litopenaeus vannamei. Shrimp were randomly allocated in 12 tanks (30 shrimp per tank) and triplicate tanks were fed with a control diet or diets containing different levels of CGA (100, 200 and 400 mg kg(-1) feed) as treatment groups. Growth performance including weight gain (WG), biomass gain (BG), feed conversion ratio (FCR), and feed intake were determined after feeding for 56 days. Antioxidant capacity were evaluated by determining the activity of total antioxidant status (TAS), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) as well as the gene expression of GSH-Px and CAT in the hepatopancreas of shrimp at the end of feeding trial and again at the end of the combined stress test. The results indicated that supplemention of CGA had no significant effects on the growth performance and the activities of TAS, SOD, GSH-Px and CAT in hepatopancreas of shrimp cultured under normal conditions for 56 days. However, compared with the control group, CGA (200, 400 mg kg(-1) feed) significantly improved the resistance of L. vannamei against the combined stress of low-salinity and nitrite, as indicated by the significant (P < 0.05) higher survival, higher activities of TAS, GSH-Px and CAT, as well as higher transcript levels of GPx and CAT gene in shrimp treated with CGA in the combined tress test. Our findings suggested that CGA possessed dual-modulatory effects on antioxidant capacity of L. vannamei and could be a potential feed additive that can enhance shrimp resistance against environmental stresses. The recommended application dosage is 200 mg kg(-1) and further studies are needed to clarify the action model of CGA efficiency.

Effect of guava leaves on growth and the non-specific immune response of Penaeus monodon.

Guava (Psidium guajava L.) leaf extracts have antiviral and antibacterial activity against shrimp pathogens such as yellow-head virus (YHV), white spot syndrome virus (WSSV), and Vibrio harveyi, which make it a potential water disinfectant for use in shrimp culture. In this study, the safety of guava leaf supplementation in shrimp was evaluated by studying its influence on growth and the non-specific immune response of Penaeus monodon. Six diets containing different levels of guava leaves (0% [basal diet], 0.025% [G1], 0.05% [G2], 0.1% [G3], 0.2% [G4], and 0.4% [G5]) were fed to groups of shrimp (1.576 ± 0.011 g body weight) in triplicate for 56 days. Growth performance (final body weight, WG, PWG, SGR) of shrimp fed guava leaf diets was significantly higher (P < 0.05) than that of shrimp fed on the basal diet. The G1 diet resulted in the highest body weight gain (308.44%), followed by the G2 (295.45%), G3 (283.05%), G5 (281.29%), G4 (276.11%), and finally the basal diet (214.58%). Survival of shrimp in the G1 diet group was higher than that of shrimp in the control and the other experimental groups; however, no statistical differences (P > 0.05) were found. Dietary supplementation with guava leaf improved the activities of prophenoloxidase (PO) and nitric oxide synthase (NOS) in serum, and of superoxide dismutase (SOD), acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LSZ) both in serum and hepatopancreas of shrimp. In the experimental groups, the activities of these enzymes followed a similar pattern of change; they increased initially at low levels of dietary supplementation and then decreased with increasing concentrations of dietary guava leaf. Serum PO and SOD activities in shrimp fed the G1 diet reached 7.50 U ml(-1) and 178.33 U ml(-1), respectively, with PO activity being significantly higher than in controls. In shrimp fed the G1 diet, SOD, ACP, and AKP activities in hepatopancreas were significantly higher than in the controls, reaching 57.32 U g(-1), 23.28 U g(-1), and 19.35 U g(-1) protein, respectively. The highest activities of serum ACP, AKP, LSZ, and of hepatopancreas LSZ, were observed in the G3 diet group. Total nitric oxide synthase (TNOS) activity was highest (64.80 U ml(-1)) in the G4 diet group, which was significantly higher than that observed in the control group. These results suggest that dietary guava leaf supplementation could enhance the growth performance and non-specific immune response of P. monodon. Therefore, guava leaf is considered safe for use as a water disinfectant in shrimp culture.

Dietary supplementation of honeysuckle improves the growth, survival and immunity of Penaeus monodon.

Two trials were conducted to determine the effects of honeysuckle on shrimp, Penaeus monodon, first on growth performance, secondly on the immune response of shrimp. In trial 1, shrimp (mean initial wet weight about 3.02 g) were fed with five diets containing 0% (basal diet), 0.1%, 0.2%, 0.4% and 0.8% honeysuckle in triplicate for 60 days. Growth performance (final body wet weight, FBW; weight gain, WG; biomass gain, BG) of shrimp fed honeysuckle diets were higher (P < 0.05) than that of shrimp fed the basal diet, shrimp fed 0.4% honeysuckle diet showed the highest value of growth performance. Shrimp fed 0.2% honeysuckle diet showed highest value of survival. The total antioxidant status (TAS) and glutathione peroxidase (GSH-Px) activity of shrimp fed 0.2%, 0.4% and 0.8% honeysuckle diets were higher (P < 0.05) than those of shrimp fed basal and 0.1% honeysuckle diets. Hepatopancreas malondialdehyde (MDA) of shrimp fed honeysuckle diets were lower (P < 0.05) than that of shrimp fed the basal diet. Total haemocyte count of shrimp fed the basal diet was lower (P < 0.05) than that of shrimp fed honeysuckle diets. Haemolymph clotting time of shrimp had the opposite trend with the total haemocyte count of shrimp. In trial 2, the shrimp were exposed to air during a simulated live transportation for 36 h after the rearing trial. The antioxidant responses were characterized by lower TAS and higher antioxidant enzyme activities (superoxide dismutase: SOD, GSH-Px) and higher oxidative stress level (MDA) in the hepatopancreas compared to levels found in trial 1. No mortalities were observed in any diet groups after 36 h of simulated live transportation. The glutathione (GSH) content and TAS of shrimp fed 0.2%, 0.4% and 0.8% honeysuckle diets were higher (P < 0.05) than those of shrimp fed the basal and 0.1% honeysuckle diets. The SOD activity of shrimp fed the basal diet was higher (P < 0.05) than that of shrimp fed honeysuckle diets. The GSH-Px activity of shrimp fed the basal diet was lower (P < 0.05) than that of shrimp fed 0.2%, 0.4% and 0.8% honeysuckle diets but without significant difference (P > 0.05) with shrimp fed 0.1% honeysuckle diet. Moreover, the oxidative stress level (MDA) recorded in the hepatopancreas with shrimp submitted to the honeysuckle diets were lower. In conclusion, results suggested that dietary intake containing honeysuckle could enhance the growth performance of P. monodon and improve its resistance to air exposure during simulated live transportation. Considering the effect of honeysuckle on both growth performance and survival of P. monodon, the level of honeysuckle supplemented in the diet should be between 0.2% and 0.4%.

Dietary values of astaxanthin and canthaxanthin in Penaeus monodon in the presence and absence of cholesterol supplementation: effect on growth, nutrient digestibility and tissue carotenoid composition.

Penaeus monodon (mean initial wet weight 1·19 (SE 0·01) g) were fed seven diets in triplicate: a control diet (D1) without carotenoids; three diets formulated to supply 0·1 % astaxanthin alone (D2), 0·2 % astaxanthin alone (D3), and a combination of 0·1 % astaxanthin and 1 % cholesterol (D4); three diets with 0·07 % canthaxanthin alone (D5), 0·13 % canthaxanthin alone (D6), and a combination of 0·07 % canthaxanthin and 1 % cholesterol (D7). Weight gain (WG, %), specific growth rate (SGR, %/d) and survival were chosen as parameters of shrimp growth performance. Total antioxidant status (TAS), superoxide dismutase (SOD), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were chosen as indices of shrimp plasma antioxidant capacity. Meanwhile, digestibility, retention efficiency and tissue carotenoids were also investigated to determine the additive effect of cholesterol on the efficiency of astaxanthin and canthaxanthin. After 74 d rearing, WG and SGR of shrimp fed D2-D4 and D7 were higher than those of shrimp fed D1 (P < 0·05). Shrimp fed D4 had the highest survival. The apparent digestibility coefficients (ADC) of astaxanthin in D2-D4 were higher than those of canthaxanthin in D5-D7 (P < 0·05). Although ADC of astaxanthin were quite high (>98 %) in D2-D4 and no differences were found among them (P>0·05), the carotenoid retention efficiencies in the whole body, muscle and shell (D2-D3 treatments) were considerably low; however, cholesterol supplementation significantly improved the carotenoid retention efficiencies in the whole body, muscle and shell (D4 treatment). Accordingly, the addition of cholesterol also significantly enhanced the carotenoid contents of tissues. Shrimp fed supplemented carotenoid diets (D2-D7) had higher TAS and lower SOD, ALT and AST than shrimp fed D1 (P < 0·05). A low dissolved oxygen stress test was conducted for 7 d after the rearing trial and shrimp survival was also compared among the treatments. The survival of shrimp fed the diets supplemented with astaxanthin or canthaxanthin was higher than that of shrimp fed D1 during the stress test (P < 0·05). In conclusion, all data suggested that astaxanthin was better than canthaxanthin as the dietary carotenoid source in the commercial diet of P. monodon, and the supplement of cholesterol could positively enhance the efficiency of astaxanthin and canthaxanthin.

Identification and expression profile of Delta-GST in black tiger shrimp (Penaeus monodon) exposed to aflatoxin B1 (AFB1).

Delta GST is an insect-specific class and a prominent class of the glutathione S-transferases family that is involved in xenobiotic detoxification and antioxidant defense. The full-length complementary DNA of delta-class GST from Penaeus monodon (PmDeltaGST; 839 bp long with a 657 bp coding region) was cloned. The encoded polypeptide of 218 amino acids had a predicted molecular mass of 24.30 kDa. Sequence homology and phylogenetic analysis showed that PmDeltaGST was significant similarity to GST genes in crustaceans and insects. Tissue expression profile analysis by quantitative real-time reverse-transcription polymerase chain showed that PmDeltaGST was constitutively expressed in all the examined tissues, with the highest expression in hepatopancreas and intestine and the weakest expression in ovary. PmDeltaGST messenger RNA expression and protein levels in hepatopancreas was significantly increased at 14 days postexposure of aflatoxin B1 (AFB1), keeping on the high level at 28 days, but decreased at 56 days. The results suggested that PmDeltaGST was involved in the response to AFB1 exposure.

Excess dietary cholesterol may have an adverse effect on growth performance of early post-larval Litopenaeus vannamei.

One experiment was conducted to determine the nutritive value of cholesterol for post-larval shrimp, Litopenaeus vannamei. Four isoenergetic and isonitrogenous diets supplemented with four levels of cholesterol (D1, D2, D3 and D4 with 0, 0.5%, 1% and 2% cholesterol, respectively) were fed to triplicate groups of L. vannamei shrimp (mean initial wet weight 0.8 mg) for 27 days. After the trial, shrimp fed the D1 diet had the best growth performance (final body weights: FBW; weight gain: WG; specific growth rate: SGR), while there was no significant difference between diet treatments with respect to survival. The whole body crude protein level in the shrimp decreased with the increase in dietary cholesterol levels, while the whole body crude lipid level in shrimps in the D4 diet treatment was significantly higher (P < 0.05) than in other diet treatments. Dietary analysis indicated that the D1 diet contained 0.92% cholesterol prior to supplementation, which may have satisfied the dietary cholesterol requirement of post-larval L. vannamei; excess dietary cholesterol may thus lead to adverse effects on the growth performance of post-larval shrimp.