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1.
The sea cucumber Apostichopus japonicus is an economic species mainly distributed along the coast of northern China, south‐eastern Russia, Japan, Republic of Korea and Democratic People's Republic of Korea. Aquaculture industry of A. japonicus has been facing severe challenge of high temperature. In this study, we studied the mRNA expression profiles of eight key metabolic enzymes involved in carbohydrate and amino acids metabolism in A. japonicus under heat stress. The expression of hexokinase, pyruvate kinase and malate dehydrogenase showed downregulated response to heat stress, while the expression of glutamate dehydrogenase, alanine transaminase and branched‐chain aminotransferase showed upregulated response. In addition, the expression of lactate dehydrogenase and glutamate synthase showed no significant difference. We also applied 1H NMR‐based metabolomics to investigate metabolic changes in the intestine tissue of A. japonicus under heat stress, the results of which revealed nine increased and 10 decreased metabolites in the heat stress group. These response genes and metabolites have potential to become markers for identifying severity of heat stress. More importantly, our findings suggest significant links between gene expression and metabolites changing, highlighting regulation networks of carbohydrate and amino acid metabolism in heat‐stressed A. japonicus.  相似文献   

2.
The effects of fulvic acid (FA) on survival and immune‐related gene expression were investigated in Litopenaeus vannamei challenged with Vibrio parahaemolyticus by immersion. Shrimp were fed with different dietary FA concentrations (1, 2, 4 and 6 g/kg feed) for 20 days (first bioassay) or 8 days (second bioassay, 2 g/kg feed of FA added every 2 days) and then challenged with V. parahaemolyticus. In a third bioassay, the expression of three immune‐related genes (translationally controlled tumour protein [TCTP], superoxide dismutase [SOD] and heat‐shock protein 70 [HSP70]) in haemocytes or hepatopancreas of experimental shrimp was measured by real‐time quantitative PCR at 0, 6, 12, 24, 48, 72 and 96 hr after FA (2 g/kg feed) administration. Fulvic acid increased survival at a concentration of 2 g/kg feed supplied every two days. Interestingly, TCTP gene expression was upregulated, whereas gene expression of SOD and HSP70 was downregulated. In conclusion, dietary fulvic acid improves survival in white shrimp challenged with V. parahaemolyticus and modulates the immune response. Therefore, FA merits further evaluation as prophylactic treatment in commercial shrimp farms.  相似文献   

3.
Using Hsp70 as a biomarker, thermal stress impinges on reproductive organs, ovary and hepatopancreas were being analyzed by determining the expression of Hsp70 mRNA inside the organs after the adult inter‐molt females were subjected to thermal treatment at 35, 30 and 28°C (Control). Results showed the expression of Hsp70 mRNA under thermal treatment of 35°C after 2 hr recovery in ovary were upregulated at 2, 4, 6, 12, 24 hr and 30 days compared to control whereas in hepatopancreas under similar treatment, the expression of Hsp70 mRNA were significantly higher than control at 6, 24 hr and 30 days. Frequency of reproductive molt at 35°C showed the ovary of females were failed to develop and only entered common molt along three consecutive molt cycles. For 30°C thermal treatment, the expression of Hsp70 mRNA was significantly higher than control after 2 hr recovery but returned to normal afterwards until 30 days’ thermal treatment. Maternal heat shock for 2 hr at 35°C were found to give significantly lower frequency of reproductive molt and longer duration of ovarian development and incubation period whereas maternal heat shock for 2 hr at 30°C gave lower frequency of reproductive molt, slower development of embryo and lower hatching success compared to untreated control. This study suggests that short and long‐term thermal stress at 30 and 35°C were found to affect the induction of Hsp70 mRNA in reproductive organs of Macrobrachium rosenbergii and also influence their reproductive performance.  相似文献   

4.
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5.
A fatty acid‐binding protein (FABP) gene designated as MnFABP10 was cloned and characterized from the freshwater prawn Macrobrachium nipponense. The full‐length cDNA of MnFABP10 was 646 bp encoding a 130 amino acid. Real‐time quantitative RT‐PCR showed that the MnFABP10 gene was expressed in various tissues with the highest expression in the hepatopancreas. The MnFABP10 mRNA levels in the hepatopancreas and ovary of M. nipponense were dependent on the stages of ovarian development. Western blot results revealed a single immunoreactive band with an estimated molecular mass of approximate 14 kDa in the developmental ovary. Then, M. nipponense with an initial body weight of 0.090 ± 0.0010 g were fed with four isonitrogenous and isocaloric diets with different oils, that is, beef tallow (BT), soybean oil (SO), pollack fish oil (FO) and a mixture of fish oil and soybean oil (FO/SO 2 : 1 w/w) for 52 days. The mRNA levels of MnFABP10 in the hepatopancreas were influenced by different lipid sources, with a peak expression observed in prawns fed SO. This study suggests that MnFABP10 may have a putative function in ovary maturation, and its mRNA expression in the hepatopancreas can be regulated by the source of dietary lipids in M. nipponense.  相似文献   

6.
In this study, we evaluated the growth, osmoregulation and energy metabolism of the oriental river prawn, Macrobrachium nipponense, reared during 6 weeks with different salinities (0, 8, 14 and 22 g/L). The results showed that the haemolymph osmolality of M. nipponense increased with an increase in ambient osmotic pressure; the isosmotic point was 490 mOs/kg H2O. The prawns showed a higher survival rate, weight gain rate and hepatopancreas index in salinity 14 g/L. Digestive enzymes were all affected by salinity, and the highest activities were observed in the salinity 14 g/L. The mRNA expression of Na+‐K+‐ATPase in gills and p53 in hepatopancreas were the highest in salinity 22 g/L. The expressions of heat shock protein 90 and glutathione S‐transferase genes in hepatopancreas were significantly higher in the salinity 8 g/L. Lipid metabolism‐related genes in hepatopancreas were significantly expressed in the salinity 14 g/L. The glucose‐6‐phosphatase gene in hepatopancreas was highly expressed in the salinity 8 and 22 g/L, and the expression of the ecdysone receptor gene in hepatopancreas was significantly higher in the salinity 14 g/L. The results showed that salinity 14 g/L could promote the growth of M. nipponense. However, higher salinity conditions may cause physiological damage, which provides a theoretical basis for brackish water culture of M. nipponense.  相似文献   

7.
In recent years, more and more attentions have been paid to the development and application of probiotics in aquaculture, and viable probiotics have been extensively studied, while rare information was available about inactivated probiotics in aquaculture. Therefore, in this study, a feeding trial was designed to investigate the effect of dietary supplementation of heat‐inactivated probiotic Bacillus clausii DE5 on growth performance, immune response and key immune genes expression in head kidney and intestine in grouper Epinephelus coioides. Fish were fed for 60 days with control diet (C) and two experimental diets containing 1.0 × 108 CFU/g live (T1) and heat‐inactivated (T2) B. clausii DE5, respectively. The probiotic treatments did not affect the final weight, weight gain (WG) and specific growth rate (SGR) of E. coioides at days 30 and 60 (p > .05), while both heat‐inactivated and live B. clausii DE5 significantly decreased the feed intake and feed conversion ratio (FCR) at day 60 (p < .05). Serum lysozyme activity and complement C3 level in the two probiotic treatments were significantly higher than those in the control (p < .05). The lysozyme activity and complement C3 level at day 60 were significantly higher than those at day 30 (p < .05), while no significant interaction effect between diet and administration date was observed. Moreover, the heat‐inactivated B. clausii DE5 significantly improved the expression of TLR5, pro‐inflammatory cytokines (IL‐8 and IL‐1β) and TGF‐β1 in head kidney and intestine (p < .05), while the live probiotic did not show any significant effect on the expression of these key immune‐related genes in head kidney and intestine. These results indicate that dietary supplementation of heat‐inactivated B. clausii DE5 effectively improved feed utilization and both the local and systemic immune responses of E. coioides.  相似文献   

8.
A 3‐hr experiment was conducted to investigate the effects of dietary myo‐inositol (MI) supplementation on survival, immune response and antioxidant abilities in Litopenaeus vannamei under acute hypoxia stress. Six practical diets were formulated with supplementation of graded levels (control group 0, 0.1, 0.2, 0.4, 0.8 and 1.6 g/kg dry diet) of MI and were randomly assigned to triplicate groups of L. vannamei (mean weight 0.40 ± 0.00 g) for 8 weeks. Ten healthy shrimp (final mean weight approximately 11–14 g) randomly selected from each tank were exposed to hypoxia stress after feeding trial. After 3‐hr acute hypoxia stress, survival of shrimp fed MI‐supplemented diets (except 0.1 and 0.4 g/kg diets) was significantly increased compared with the control group. Shrimp fed control diet had lower activities of alkaline phosphatase (AKP), acid phosphatase (ACP), total antioxidant capacity (T‐AOC) and glutathione peroxidase (GPX), and higher malondialdehyde (MDA) and protein carbonyl (PC) contents in hepatopancreas than those fed the MI‐supplemented diets. In addition, mRNA expression levels of heat shock protein 70 (Hsp70), catalase (CAT) and penaeidin were significantly differentially regulated in hepatopancreas. In summary, dietary MI supplementation may have a positive effect on improving resistance to acute hypoxia stress of L. vannamei.  相似文献   

9.
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10.
The system nitrogen (N), phosphorous (P) budget, N/P utilization rate and the physiological response mechanism of the abalone Haliotis discus hannai (body weight: 12.87 ± 0.82 g) and the sea cucumber Apostichopus japonicus (body weight: 10.85 ± 1.16 g) to different co‐culture environment conditions were examined. Animals were kept in a multilayer, cubic recirculating aquaculture system at different polyculture densities (abalones at 400 ind/m2in monoculture [Group C] and abalones at 400 ind/m2with sea cucumbers at 10 ind/m2 [AS1] or 20 ind/m2 [AS2]). Each treatment was replicated four times, and the experimental cycle was 90 days. No significant difference in survival rate of abalones was detected when the stocking density of sea cucumbers increased from 10 to 20 ind/m2, but the concentrations of total ammonia nitrogen and nitrite nitrogen in the water were significantly higher in AS2 than in AS1. Survival rate, specific growth rate (SGR) of body weight of sea cucumbers and SGR of body weight of abalones were significantly lower in AS2 than in AS1. No significant difference in protease (PES), lipase, amylase and cellulase activities of abalones was identified between Group C and AS1, but the PES and amylase activities of abalones and sea cucumbers in AS1 were significantly higher than those in AS2. In AS1, the N/P output from harvesting of abalones and sea cucumbers and the N/P utilization rates were significantly higher than those in AS2. Although the N/P output from faeces was significantly lower in AS2 than in Group C and AS1, the N/P output from the water layer was significantly higher than that in AS1. The expression levels of Cu/Zn superoxide dismutase (Cu/Zn‐SOD) and heat shock protein 70 (HSP70) of both abalones and sea cucumbers in AS2 were significantly higher than those in AS1. No significant difference in expression of catalase (CAT) and HSP90of abalones was identified among these groups, but the expression levels of CAT and HSP90of sea cucumbers in AS2 were significantly higher than those in AS1. These results indicate that stocking sea cucumbers at 10 ind/m2 in the polyculture system will relieve the organic load on the system and improve the N/P utilization rate. It will also increase aquaculture production and improve the ecological and economic benefits of the system.  相似文献   

11.
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12.
This study deals with the mortality and related physiological responses of aestivating sea cucumber Apostichopus japonicus to acute salinity decrease. Aestivating and active sea cucumbers were exposed to a decrease in salinity (from 30 to 20 psu) at a rate of 2.5 psu every 6 h, and then maintained at 20 psu for 96 h. The mortality of aestivating sea cucumbers was ~30%, which was significantly higher than that of active sea cucumbers (~10%). This result indicated that sea cucumbers in aestivation were more susceptible to hypo‐salinity stress. To elucidate the underlying physiological mechanisms, the osmotic pressure in coelomic fluid and the levels of hsp70 and hsp90 mRNA in aestivating and active sea cucumbers were measured. No significant difference in osmoregulation was observed between the two groups. The osmotic pressure of coelomic fluid in both groups changed with decrease in ambient salinity. There were significant differences in the time course and magnitude of hsp70 and hsp90 expression between the two groups. After exposure to decreased salinity, aestivating sea cucumbers showed a delayed up‐regulation of hsp70 and hsp90 expression compared with animals in active state, and these levels decreased rapidly to control values. The expression of hsp70 and hsp90 in aestivating sea cucumbers were significantly lower than those in active sea cucumbers after salinity change. The differences in hsp70 and hsp90 expression between the states may partly explain the higher mortality of sea cucumbers in aestivation when exposed to low salinity.  相似文献   

13.
The experiments explored the pharmacokinetics (PK) properties of oxolinic acid (OXA) after oral administration at three dosages (10, 30 and 80 mg/kg) via medicated feed in the shrimp. The results showed that the Cmax values of 4.31, 14.93 and 16.62 mg/L and AUC0–∞ values of 92.61, 252.30 and 364.27 mg hr?1 L?1 were observed at three OXA dosage groups in the haemolymph respectively. In the hepatopancreas, Cmax values of 7.90, 27.23 and 60.51 mg/kg and AUC0–∞ values of 42.01, 133.06 and 219.06 mg hr?1 L?1 were observed at 0.5 hr post administration respectively. In the muscle, Cmax values of 1.62, 5.80 and 7.36 mg/kg and the AUC0–∞ values of 25.64, 98.10 and 134.24 mg hr?1 L?1 were observed at 2 hr post administration respectively. In the gills, Cmax values of 2.87, 8.08 and 12.12 mg/kg and the AUC0–∞ values of 51.38, 118.65 and 206.48 mg hr?1 L?1 were observed at 4 hr post administration respectively. In addition, the in vitro MIC values of OXA at three dosages against 132 strains of Vibrio were examined and showed that the minimum inhibitory concentration (MIC) values for OXA primarily ranged from 0.15–1.25 µg/ml, including eight strains of Vibrio showing MIC values ≥5 µg/ml. The MIC50 and MIC90 values of 132 strains were 0.62 and 1.25 μg/ml respectively. The AUC0–24/MIC90 ratios of Vibrio were 140.4 in 30 mg/kg group. Furthermore, the P‐glycoprotein (P‐gp) expression was determined in shrimp tissues after administration to three dosage groups (10, 30 and 80 mg/kg). The results showed that P‐gp expression was up‐regulated in the hepatopancreas (5.36‐, 13.68‐ and 31.06‐fold respectively) compared with the control group.  相似文献   

14.
Retinoblastoma (Rb) is a multifunctional regulator involved in several key cellular processes, such as cell cycle control, cell differentiation, tumorigenesis and senescence. In this study, an Rb‐like gene, PmRBL, was cloned from black tiger shrimp (Penaeus monodon). The full‐length cDNA sequence of PmRBL is 4,069 bp with an open reading frame of 3,243 bp, which encodes 1,080 amino acids. Quantitative real‐time PCR (qRT‐PCR) analysis indicated that PmRBL was highly expressed in the gills, hepatopancreas and ovaries of P. monodon. The highest PmRBL expression levels were observed in stage III of the ovarian development in P. monodon. RNA interference experiments were conducted to examine the expression profiles of PmRBL, PmCDK2 and PmCyclin E. The knockdown of PmRBL in the ovary and hepatopancreas by dsRNA‐RBL was successful. After dsRNA‐RBL was injected into the shrimp, the relative expression levels of PmCDK2 and PmCyclin E were upregulated at 12–72 hr in the ovaries and hepatopancreas. The localization and level of PmRBL expression in the ovary and hepatopancreas were investigated through in situ hybridization, which revealed consistent results with those of qRT‐PCR. Therefore, PmRBL, PmCDK2 and PmCyclin E may be involved in vitellogenin synthesis and ovarian maturation in P. monodon.  相似文献   

15.
To determine the effects of linolenic acid (LNA, 18:3n‐3) in oriental river prawn (Macrobrachium nipponense), an 8‐week feeding experiment was conducted using six isonitrogenous and isoenergetic semi‐purified diets containing 0.07 g/kg (control), 7.3 g/kg, 16.6 g/kg, 20.2 g/kg, 27.3 g/kg and 36.3 g/kg LNA. The hepatopancreas lipid content decreased significantly when dietary LNA content was >20.2 g/kg. Fatty acid analysis revealed that the percentage of 18:3n‐3 in the hepatopancreas significantly increased with increasing dietary LNA levels, while 20:5n‐3, 22:5n‐3 and 22:6n‐3 levels in the hepatopancreas decreased in a curvilinear manner as dietary LNA increased. Additionally, qRT‐PCR results revealed that hepatopancreas mRNA expression of acetyl‐CoA carboxylase (ACC) decreased with increasing dietary LNA, while the greatest carnitine palmitoyl transferase‐1(CPT1) mRNA expression was observed in the 2.73 g/kg and 36.3 g/kg groups. Furthermore, hepatopancreas mRNA expression of acyl‐CoA delta‐9 desaturase (SCD) and fatty acyl elongase 6(elovl6) was downregulated when prawns fed the diets containing >20.2 g/kg LNA. These results indicate that dietary 18:3n‐3 could decrease lipid deposition through increased fatty acid β‐oxidation and modulated fatty acid synthesis, and alter fatty acid composition by regulating fatty acyl elongase and fatty acyl desaturase mRNA expression in the M. nipponense.  相似文献   

16.
We examined the effects of salinity stress and ammonia stress on alteration of the haemocyte count, phenoloxidase activity, expressions of immune‐related genes including prophenoloxidase (proPO), crustin, heat shock protein 70 (HSP70), and expressions of stress‐responsive neurohormone (Bur‐α and Bur‐β) in the thoracic and abdominal ganglia of giant river prawn Macrobrachium. These parameters of prawn that subjected to salinity stress (transferred from 0‰ to 5‰ and 10‰), and subjected to ammonia‐nitrogen (ammonia‐N) stress (transferred from 0 to 0.262 mg/L and 0.786 mg/L) were examined after 0, 3, 6, 24, 72 and 168 hr respectively. During the initial period of 3, 6 and 24 hr, granulocyte haemocyte (granular and semi‐granular hemocyte) count and PO activity significantly decreased, while expressions of Bur‐α and Bur‐β significantly increased. After 24 hr, granulocyte haemocyte count and PO activity significantly increased, whereas expressions of Bur‐α and Bur‐β significantly decreased. The expressions of proPO, crustin and HSP70 were significantly downregulated in the prawn that subjected to salinity stress and ammonia‐N stress at all time periods of 3–168 hr. In conclusion, changes in the granulocyte haemocyte count of M. rosenbergii following salinity stress and ammonia‐N stress are closely associated with the changes of Bur‐α and Bur‐β expressions.  相似文献   

17.
A feeding trial was conducted to study the effect of dietary lipid on growth performance and heat‐shock protein (HSP70 and HSP60) response of white seabass (WSB), Atractoscion nobilis. Five diets were formulated to contain 440 g kg?1 protein from 300 g kg?1 fish meal, 240 g kg?1 soybean meal and 100 g kg?1 soy protein concentrate with different levels of lipid: 100, 120, 140, 160 or 180 g kg?1. At the end of the trial, heat shock response based on HSP70 and HSP60 was measured in liver and white muscle from fish at ambient temperature and temperature shock conditions. Final weight and percent gain were significantly higher for fish fed the 100 g kg?1 lipid diet than for fish fed the rest of the diets (P ≤ 0.05). Feed conversion ratio was lowest for fish fed the 100 g kg?1 lipid diet. The HSP70 and HSP60 responses were positively correlated to dietary lipid levels following temperature shock. At ambient temperature, HSP60 and HSP70 responses in muscle and HSP60 response in liver increased with dietary lipid level. Temperature shock significantly increased the HSP response of fish in all treatments. Results of this study demonstrated that a moderate (110–120 g kg?1) level of dietary lipids would be recommended for production diets but a higher dietary lipid level may be required for optimal stress tolerance.  相似文献   

18.
Endocrine disrupting chemicals (EDCs) are of substantial concern when contaminating waters. We studied gene expression of larvae from river pufferfish, Takifugu obscurus exposed to EDCs. We cloned the full‐ or partial‐length cDNA sequence of genes related to the cortisol pathway, such as the glucocorticoid receptor (GR), mineralocorticoid receptor (MR), heat shock protein (HSP) 70 and 90, phosphoenolpyruvate carboxykinase (PEPCK) and the FK506‐binding protein 4 (FKBP52). The tissue and time‐dependent expression of mRNA was studied by real‐time PCR in T. obscurus exposed to two representative EDCs (bisphenol‐A and 4‐tert‐octylphenol). HSP70, HSP90, MR and FKBP52 mRNA expression level was much higher than GR and PEPCK expression. GR mRNA expression level was significantly upregulated after 48 h in both EDC‐treated fish groups compared with the control. Both groups also showed down‐regulation patterns after 48 h. The initial expression of PEPCK in both groups was down‐regulated after 24 h. In the bisphenol‐A treated group, the expression of FKBP52 showed substantially high up‐regulation from 6 to 48 h after exposure, and then strikingly reduced its expression level at 72 h. Our results provide insights into the EDCs metabolizing system of T. obscurus and offers baseline information for further research related to the possible use as a bio‐indicator of this commercially important aquaculture fish species.  相似文献   

19.
Stunted growth in pond‐reared Litopenaeus vannamei was observed in different farms located in Tamil Nadu and Andhra Pradesh, India. No mortality was associated with stunted growth. PCR assay on these samples revealed the presence of Enterocytozoon hepatopenaei (EHP) in stunted shrimp. Tissue distribution of EHP in naturally and experimentally infected shrimp was studied by PCR and histology. Histological examination revealed the presence of EHP in hepatopancreas and gut, but not in other organs. The PCR assay revealed the presence of EHP in all the organs tested in both naturally and experimentally infected shrimp. Healthy shrimp were challenged with E. hepatopenaei by intramuscular injection and oral route, and no mortality was observed in both routes after 30 days post‐challenge. Different developmental stages of the microsporidian parasite were observed in the hepatopancreatic epithelial cells. Biochemical parameters such as total protein, albumin, aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase were measured in the haemolymph of naturally and experimentally EHP‐infected shrimp. All biochemical parameters mentioned were found to be significantly higher in EHP‐infected shrimp when compared to normal shrimp. This is the first report relating AST and ALT levels to EHP infection in naturally and experimentally infected shrimp.  相似文献   

20.
The present study was to determine the time‐dependent alterations in growth performance, histomorphology, digestive enzymes activities and the genes expression related to target of rapamycin (TOR) signalling pathway involved in hepatopancreas and intestine of Litopenaeus vannamei at 3, 6, 12, 18, 24 and 30 days after Aflatoxin B1 (AFB1) challenge. The results manifested that AFB1 could induce a significant decrease in growth performance and visible variations of the hepatopancreas and intestine structures in shrimp. Meanwhile, the digestive enzymes activities and genes expression level were significantly lower in the experimental group than in the control group and showed a time‐dependent decrease (p < 0.05). The expression levels of tor and s6k were significantly higher in the experimental group than in the control group from days 6 to 24 and showed a tendency to increase first and then decrease as a whole (p < 0.05). Meanwhile, we analysed candidate pathways involved in hepatopancreas and intestine of shrimp in metabolic response to AFB1. The mainly disturbed pathways related to metabolism in hepatopancreas were involved in pyrimidine, glycosylphosphatidylinositol, glycosaminoglycan, vitamin and amino acid biosynthesis and metabolism, while the mainly disturbed metabolic pathways in intestine were glycosaminoglycan, vitamin and amino acid biosynthesis and metabolism.  相似文献   

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