Effects of dietary protein levels on the growth,survival, amylase and trypsin activities in large yellow croaker,Pseudosciaena Crocea R., larvae

A feeding trial was conducted to investigate the effects of dietary graded protein levels on the growth, survival, amylase and trypsin activities of large yellow croaker (Pseudosciaena crocea R.) larvae from 12 to 42 days after hatching (DAH). Five approximately isoenergetic microbound diets (16.65 MJ/kg diet) were formulated to contain different protein (47.1%, 52.0%, 57.1%, 62.2% and 67.5%) levels. Frozen copepods, containing 54.5% crude protein (CP), 6.0% crude lipid, 27.2% ash and 6.7% glycogen, were used as a control. Each diet was randomly fed to triplicate groups of larvae with an initial mean body weight of 1.76 ± 0.09 mg (mean ± SD) in 180 L white plastic tanks, and each tank was stocked initially with 3500 larvae. Both the survival and the specific growth rate (SGR) of large yellow croaker larvae significantly increased with increasing dietary protein level up to 57.1%, and decreased thereafter. Frozen copepods resulted in intermediate survival and low SGR compared with the other diets. Whole‐body moisture and protein of larvae were not significantly affected by the dietary protein level. In contrast, whole‐body lipid of larvae fed diet with 47.1% CP was significantly higher (P<0.05) than those from fish fed the diets containing more than 57.1% CP. Additionally, fish fed the frozen copepods had the lowest whole‐body protein and lipid. The amylase‐specific activity increased with increasing dietary carbohydrate level during the period of this experiment. However, trypsin activity was not significantly affected by the dietary protein content before 42 DAH, indicating a later onset of trypsin than amylase in the regulation of enzymatic synthesis induced by a dietary substrate.     

Changes in activities and mRNA expression of lipoprotein lipase and fatty acid synthetase in large yellow croaker,Larimichthys crocea (Richardson), during fasting

Over‐winter fasting and man‐made food deprivation to increase meat quality are common in the process of large yellow croaker (Larimichthys crocea) aquaculture. This study aimed to determine the changes in lipoprotein lipase (LPL) and fatty acid synthetase (FAS) activities and mRNA expressions, and the relationships between these changes and fat content in large yellow croaker liver and muscle tissues during fasting. A total of 2933 bp LPL cDNA, including an open reading frame of 1533 bp encoding 510 amino acids, and a 1233 bp fragment of FAS cDNA coding 411 amino acids were cloned. Expressions of both genes were ubiquitous. During a 35‐day fasting period, the hepatosomatic index and fat content in muscle and liver were significantly decreased (P < 0.05). mRNA levels of LPL increased significantly during the fasting period except the first 3 days, and FAS mRNA levels decreased significantly in both muscle and liver (P < 0.05) although there were some fluctuations. Muscle and liver LPL activities were significantly higher following fasting for 7 days, decreased to the initial value following fasting for 14 days, and elevating significantly afterwards (P < 0.05). FAS activities in muscle and liver maintained a significantly decreasing trend during the short‐term fasting (P < 0.05) and kept obviously rising thereafter (P < 0.05). Activities and mRNA levels of both LPL and FAS were not always consistent, which implied that both pre‐translational and post‐translational regulations existed during fasting. Our results suggest that the reasonable fasting time is 21 days before harvesting when fat content decreased significantly.     

Physiological and molecular changes in large yellow croaker (Pseudosciaena crocea R.) with high‐fat diet‐induced fatty liver disease

Fatty liver disease is regularly observed in cultured large yellow croaker, and the disease leads to lower growth rates and reduced harvest yields. The goal of this study was to achieve a more detailed understanding of the physiological and molecular changes in response to high‐fat diet‐induced fatty liver in large yellow croaker. Large yellow croaker fed a high‐fat diet (HFD) for 9 weeks developed hepatic steatosis characterized by histological observation and significantly increased plasma triglyceride levels and serum alanine aminotransferase (ALT) activities. However, no significant differences in serum total protein, glucose, cholesterol, non‐esterified free fatty acids (NEFA), aspartate aminotransferase, high‐density lipoprotein and low‐density lipoprotein were observed between the normal diet and the high‐fat diet (HFD) group. The fatty acid composition of tissue lipids was not affected significantly by dietary lipid levels. Gene expression analysis demonstrated that the HFD decreased fatty acid synthase expression and increased PPARγ expression, but had no effect on lipoprotein lipase and PPARα expression. These results suggest that the HFD‐induced physiological changes and fatty liver may be due to the alteration of related gene expression. As such, further investigations of the metabolic pathways and differentially expressed genes are of particular significance in the mechanistic study and understanding of HFD‐induced fatty liver disease.     

Effects of dietary tea polyphenols on growth,biochemical and antioxidant responses,fatty acid composition and expression of lipid metabolism related genes of large yellow croaker (Larimichthys crocea)

The study was conducted to investigate the effects of dietary tea polyphenols (TP) on growth performance, biochemical and antioxidants responses, fatty acid composition, and lipid metabolism‐related gene expressions of large yellow croaker (Larimichthys crocea). Four diets were formulated with different levels of TP (0.00%, 0.01%, 0.02% and 0.05%). Results showed that growth performance of L. crocea were not different among dietary treatments. Compared with the control group, fish in 0.02% TP group had lower body and hepatic lipid content and lower total cholesterol content. The minimum content of triglycerides and low‐density lipoprotein‐cholesterol were found in 0.05% TP group. Hepatic n‐6 PUFA and n‐3 PUFA were significantly higher in TP supplementation groups. Malondialdehyde content was lower in TP supplementation groups, and superoxide dismutase activity was higher in 0.01% TP group than the control group. The mRNA expressions of carnitine palmitoyltransferase1, acyl‐CoA oxidase and peroxisome proliferators‐activated receptor α were up‐regulated in 0.01% and 0.02% TP groups, while lipoprotein lipase expression was down‐regulated in TP supplementation groups than the control group. Results suggested that 0.01%–0.02% TP supplementation could reduce the deposition of liver lipid of L. crocea caused by high‐lipid diet, which might be due to the increase in lipid oxidation related gene expressions.     

Optimal dietary lipid level for large yellow croaker (Pseudosciaena crocea) larvae

A 30‐day feeding experiment was conducted in blue tanks (70 × 50 × 60 cm, water volume 180 L) to determine the effects of dietary lipid levels on the survival, growth and body composition of large yellow croaker (Pseudosciaena crocea) larvae (12 days after hatchery, with initial average weight 1.93 ± 0.11 mg). Five practical microdiets, containing 83 g kg^?1 (Diet 1), 126 g kg^?1 (Diet 2), 164 g kg^?1 (Diet 3), 204 g kg^?1 (Diet 4) and 248 g kg^?1 lipid (Diet 5), were formulated. Live feeds (Artemia sinicia nauplii and live copepods) were used as the control diet (Diet 6). Each diet was randomly assigned to triplicate groups of tanks, and each tank was stocked with 3500 larvae. During the experiment, water temperature was maintained at 23(±1) °C, pH 8.0 (±0.2) and salinity 25 (±2) g L^?1. The results showed that dietary lipid significantly influenced the survival and growth of large yellow croaker larvae. Survival increased with the increase of dietary lipid from 83 to 164 g kg^?1, and then decreased. The survival of larvae fed the diet with 83 g kg^?1 lipid (16.1%) was significantly lower than that of larvae fed other diets. However, the survival in larvae fed the diet with 16.4 g kg^?1 lipid was the highest compared with other artificial microdiets. Specific growth rate (SGR) significantly increased with increasing dietary lipid level from 83 to 164 g kg^?1 (P < 0.05), and then decreased. The SGR in larvae fed the diet with 164 g kg^?1 lipid (10.0% per day) was comparable with 204 g kg^?1 lipid (9.6% per day), but were significantly higher than other microdiets (P < 0.05). On the basis of survival and SGR, the optimum dietary lipid level was estimated to be 172 and 177 g kg^?1 of diet using second‐order polynomial regression analysis respectively.     

Influence of dietary lipid on growth performance and some lipogenesis‐related gene expression of tongue sole (Cynoglossus semilaevis) larvae

A 30‐day feeding trial was conducted to investigate the effects of dietary lipid levels on growth performance, activities of digestive enzymes, fatty acid composition and some lipogenesis‐related gene expression of half‐smooth tongue sole (Cynoglossus semilaevis) larvae. Five isoproteic diets were formulated with graded lipid levels (6.68%, 9.84%, 13.47%, 17.89% and 21.88% dry weight) using fish oil as the main lipid source. Each diet was randomly allocated to triplicate groups of 150 larval tongue sole (35 DAH, 54 ± 1 mg). Fish were fed five times daily to apparent satiation during the feeding experiment. Results showed that, the survival rate (SR) of larvae increased significantly firstly, and thereafter decreased significantly. The specific growth rates (SGR) of larvae fed the diet with 13.47% lipid were significantly higher than other treatments. Larvae fed 9.84% or 13.47% dietary lipid showed higher trypsin, lipase, leucine aminopeptidase and alkaline phosphatase (AP) activities than other treatments, whereas amylase activity nearly showed reverse trend with them. The fatty acid composition of the tongue sole larvae was well correlated with dietary fatty acid profile. Expression of acetyl‐CoA carboxylase alpha (ACC1) was found to be slightly negatively correlated with dietary lipid level, and high dietary lipid level depressed the expressions of acetyl‐CoA carboxylase beta (ACC2) and fatty acid synthase (FAS) mRNA expression significantly, implying that larvae may cope with high dietary lipid mainly through down‐regulating lipogenesis‐related gene expression of FAS and ACC2. Besides, on the basis of SGR, the optimal dietary lipid level for larval tongue sole was estimated to be 13.56% using second‐order polynomial curve.     

Ontogenic development of enzymatic activity and digestive system in Jullien's golden carp (Probarbus jullieni Sauvage, 1880)

Ontogenic development of the main enzymes and histological structure of digestive organs were studied in Jullien's golden carp (Probarbus jullieni) from hatching until 50 days after hatching (DAH). The larval fish were produced by artificial insemination and fed only Moina sp. till end of experiment. Body weight (mg) and total length (cm) of Jullien's golden carp increased exponentially and linearly. The results indicate the fish weight grew fast with increasing rate, while length increased at a constant rate over the studied period. Up‐regulation of acid protease was observed in newly hatched larvae and the specific activity gradually decreased with time. Trypsin specific activity was relatively stable within the first 35 DAH, while fluctuations in chymotrypsin were observed. Among these three proteolytic enzymes, acid proteases exihibited relatively high specific activity in newly hatched larvae, suggesting a role in yolk protein degradation. Alkaline proteases became more prominent with age and correlated with an abrupt decrease in acid proteases. Increased lipase‐specific activity appeared within 3 DAH and then gradually decreased with time, indicating the capacity to digest yolk lipid reserve. Amylase and cellulase‐specific activities changed in a similar manner, and the sensitivity to time was higher in amylase than in cellulase. The digestive organs and accessory organs developed around 3–5 DAH. However, intestinal histology was almost fully developed around 18 DAH. These findings should be useful for deciding the preferred timing for weaning, as well as on developing artificial diets referenced to the physiological changes of digestive enzymes.     

Feeding selectivity in early life stages of Rhamdia voulezi under experimental conditions

The feeding selectivity of laboratory‐reared larvae of Rhamdia voulezi was assessed to investigate the preferred preys in early life stages. Three experiments were conducted at different larval development stages, starting soon after the absorption of the yolk sac (on fourth day after hatching – DAH), using zooplankton from fish ponds as food. Differences were detected in the patterns of prey selection among development stages. At first, the larvae prefer small preys, such as rotifers and cladocerans. Later, when they are more developed, they prefer to feed on large preys, such as copepods. On the fourth DAH, larvae in the pre‐flexion and initial flexion stage strongly selected rotifers and cladocerans (Diaphanosoma spinulosum, D. brevireme, Moina sp., M. micrura and M. minuta). At the other stages, cladocerans continued to be strongly selected. However, on the eighth DAH, larvae in flexion stage selected less strongly copepods Argyrodiaptomus azevedoi, Metacyclops mendocinus and Termocyclops decipiens. On the tenth DAH, larvae in postflexion stage selected more strongly copepods Argyrodiaptomus furcatus, Notodiaptomus cf. spinuliferus and M. mendocinus. As larvae grow, they tend to specialize in feeding of a particular prey. Prey selection of R. voulezi larvae was based both on species and zooplankton size. The size of the zooplankton ingested was related to the mouth gape size of larvae.     

Effects of stocking density on lipid deposition and expression of lipid-related genes in Amur sturgeon (<Emphasis Type="Italic">Acipenser schrenckii</Emphasis>)

To investigate the correlation between lipid deposition variation and stocking density in Amur sturgeon (Acipenser schrenckii) and the possible physiological mechanism, fish were conducted in different stocking densities (LSD 5.5 kg/m³, MSD 8.0 kg/m³, and HSD 11.0 kg/m³) for 70 days and then the growth index, lipid content, lipase activities, and the mRNA expressions of lipid-related genes were examined. Results showed that fish subjected to higher stocking density presented lower final body weights (FBW), specific growth ratio (SGR), and gonad adipose tissue index (GAI) (P < 0.05). Lower lipid content was observed in the liver, gonad adipose tissue and muscle in sturgeons held in HSD group (P < 0.05). The serum concentrations of triglyceride (TG), total cholesterol (TC), and high-density lipoprotein cholesterol (HDL-C) decreased significantly with increasing stocking density, while no significant change was observed for low-density lipoprotein cholesterol (LDL-C). Furthermore, the cDNAs encoding lipoprotein lipase (LPL) and hepatic lipase (HL) were isolated in Amur sturgeon, respectively. The full-length LPL cDNA was composed of 1757 bp with an open reading frame of 501 amino acids, while the complete nucleotide sequences of HL covered 1747 bp encoding 499 amino acids. In the liver, the activities and mRNA levels of LPL were markedly lower in HSD group, which were consistent with the variation tendency of HL. Fish reared in HSD group also presented lower levels of activities and mRNA expression of LPL in the muscle and gonad. Moreover, the expressions of peroxisome proliferator-activated receptor α (PPARα) in both the liver and skeletal muscle were significantly upregulated in HSD group. Overall, the results indicated that high stocking density negatively affects growth performance and lipid deposition of Amur sturgeon to a certain extent. The downregulation of LPL and HL and the upregulation of PPARα may be responsible for the lower lipid distribution of Amur sturgeon in higher stocking density.     

普安银鲫胚胎发育中脂质代谢酶基因的表达及葡萄糖的影响

采用荧光定量PCR技术检测了普安银鲫胚胎发育中乙酰辅酶A羧化酶α、脂肪酸合成酶、脂蛋白脂酶和肝脂酶基因表达的特点及葡萄糖浸泡的影响,以探讨乙酰辅酶A羧化酶α、脂肪酸合成酶、脂蛋白脂酶和肝脂酶基因在普安银鲫胚胎发育中的表达及mRNA表达水平及葡萄糖对这4种基因mRNA表达量的影响。试验结果显示,乙酰辅酶A羧化酶α、脂肪酸合成酶、脂蛋白脂酶和肝脂酶基因在普安银鲫的成熟卵期均有表达,且随胚胎发育而上升。在15g/L葡萄糖处理组中,乙酰辅酶A羧化酶α、脂肪酸合成酶、脂蛋白脂酶和肝脂酶从原肠期至出膜前期mRNA表达量升高。在原肠期、晶体出现期和出膜前期,葡萄糖能显著上调脂肪酸合成酶和乙酰辅酶A羧化酶αmRNA的表达(P0.05)。在原肠期,葡萄糖能稍上调脂蛋白脂酶和肝脂酶mRNA的表达,但差异不显著(P0.05)。在晶体出现期和出膜前期,葡萄糖能显著上调脂蛋白脂酶和肝脂酶mRNA的表达(P0.05)。研究表明,乙酰辅酶A羧化酶α、脂肪酸合成酶、脂蛋白脂酶和肝脂酶属母源性酶类,其合子基因在原肠期已经开始表达。适宜水平的葡萄糖促进脂肪酸合成酶和乙酰辅酶A羧化酶αmRNA的表达有利于脂肪酸合成酶和乙酰辅酶A羧化酶α的积累,为仔鱼出膜做准备;促进脂蛋白脂酶和肝脂酶mRNA表达有利于脂质的分解供能。     

Digestive enzyme activity of the red porgy (Pagrus pagrus, L.) during larval development under culture conditions

The ontogenic development of the main digestive enzymes (proteases, amylase and lipase) in the red porgy, Pagrus pagrus, larvae was assayed during the larval development. The green water technique was carried out for larval rearing and whole‐body homogenates were used for enzymatic assays in triplicate. Significant alterations in specific activities of all digestive enzymes measured during the period of this study were mostly related to metamorphosis and weaning. Trypsin‐ and chymotrypsin‐specific activities were first detected on day 3, together with opening of the mouth, and slightly increased until 25 days after hatching (DAH). After this period, the specific activities of these enzymes slightly decreased. Pepsin was first detected on day 28, concurrent with stomach formation, and a sharp increase was observed until 30 DAH. A slight decrease was measured from this date until the end of the experiment. Both amylase and lipase were measured for the first time on days 2 and 4 respectively, and the specific activities of these enzymes showed similar patterns during the first week of the study. Then, slight variations were observed until 30 DAH and while lipase‐specific activity declined, an increase in the specific activity of amylase was found until the end of the experiment. It is concluded that the variations observed in the specific activity of digestive enzymes were related to either metamorphosis, such as the formation of the stomach (28 DAH), or to changes in food composition. The profile of the developmental pattern of the main digestive enzymes detected in P. pagrus is similar to that described for other Sparid species.     

Short‐term starvation in silver pomfret (Pampus argenteus): molecular effects on lipid mobilization and utilization

To investigate the effects of short‐term starvation on lipid metabolism, juvenile silver pomfret (Pampus argenteus) with an initial weight of 18.1 ± 1.53 g were starved for 6 days. Fish were sampled on days 0, 2, 4 and 6 of starvation (S0, S2, S4 and S6, respectively) and then analysed. The results showed that short‐term starvation induced a significant decrease in visceral index, hepatosomatic index. The crude lipid content was also significantly decreased by 42.22% and 82.96% on S2 and S6 respectively. In addition, short‐term starvation significantly decreased the plasma triacylglycerol, non‐esterified fatty acid and low‐density lipoprotein cholesterol. Moreover, short‐term starvation significantly increased the expressions of hormone‐sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) at the mRNA and protein levels in the muscle and the ATGL expression at the protein level in the liver. The expressions of fatty acid translocase (CD36) and plasma membrane fatty acid‐binding protein (FABPpm) at the protein level in the muscle were significantly increased by short‐term starvation, while the expressions of fatty acid transport protein 1 (FATP1), FABPpm and lipoprotein lipase (LPL) at the protein level in the liver were significantly increased by short‐term starvation. Furthermore, the expressions of peroxisome proliferator activated receptor (PPAR) α and PPARγ at the mRNA and protein levels in the muscle were significantly elevated by short‐term starvation. These findings suggested that short‐term starvation increased lipid mobilization and utilization possibly through activation of lipolysis‐related genes (HSL and ATGL), lipid uptake‐related genes (LPL, CD36, FATP1 and FABPpm) and PPARs.     

Effects of Dietary Carbohydrate‐to‐lipid Ratio on Growth Performance,Body Composition,Digestive Enzyme Activities,and Hepatic Enzyme Activities in Juvenile Large Yellow Croaker,Larimichthys crocea

An 8‐wk feeding trial was conducted to investigate the effects of dietary carbohydrate‐to‐lipid ratios (CHO : L) on growth performance, body composition, digestive enzyme activities, and hepatic enzyme activities of juvenile large yellow croaker, Larimichthys crocea. Six isonitrogenous (45% crude protein) and isoenergetic (18 kJ/g gross energy) diets with varying CHO : L ratios (0.07, 0.48, 1.20, 2.19, 4.81, and 10.48) were fed to triplicate groups of large yellow croaker in floating sea cages. Results showed that the highest specific growth rate (SGR) was found in fish fed diets with CHO : L ratio of 2.19. Fish fed the lower (0.07 and 0.48) CHO : L ratios tended to produce lower growth (P < 0.05). The whole‐body lipid content significantly decreased, while hepatosomatic index, liver glycogen content, and plasma glucose concentration significantly increased as dietary CHO : L ratios increased (P < 0.05). Plasma total cholesterol, triglyceride, and low‐density lipoprotein cholesterol concentrations significantly decreased with elevated dietary CHO : L ratios (P < 0.05). The increasing dietary CHO : L ratios significantly stimulated the activities of intestinal amylase and hepatic pyruvate kinase and depressed the activity of hepatic phosphoenolpyruvate carboxykinase (P < 0.05). Based on a second‐order polynomial regression analysis of SGR, 2.38 was determined as the optimal dietary CHO : L ratio for juvenile large yellow croaker.     

Effects of dietary phospholipid level in cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis>) larvae: growth,survival, plasma lipids and enzymes of lipid metabolism

A study was conducted to determine the effects of dietary phospholipid (PL) levels in cobia (Rachycentron canadum) larvae with regard to growth, survival, plasma lipids and enzymes of lipid metabolism. Fish with an average weight of 0.4 g were fed diets containing four levels of PL (0, 20, 40 and 80 g kg⁻¹dry matter: purity 97%) for 42 days. Final body weight (FBW), weight gain (WG) and survival ratio were highest in the 8% PL diet group and mortality was highest in PL-free diet group. We examined the activities of lipoprotein lipase (LPL) and hepatic lipase (HL) in liver, lecithin-cholesterolacyltransferase (LCAT) in plasma as well as plasma lipids and lipoprotein. LCAT activity showed a decrease of more than two-fold in PL-supplemented diet groups compared with the PL-free diet group. HL activity was highest in the 8% PL diet group and the other three groups showed no difference. LPL activity was significantly higher in the PL-supplemented diet groups than in the PL-free diet group. The dietary intervention significantly increased plasma phospholipids and total cholesterol (TC) levels, and the higher free cholesterol (FC) level contributed to the TC level. However, the fish fed PL exhibited a significantly decreased plasma triglyceride (TG) level. The lipoprotein fractions were also affected significantly by the PL. The PL-supplemented diet groups had significantly higher high-density lipoprotein (HDL) compared with the PL-free diet group, but showed a marked decrease in very low-density lipoprotein (VLDL). The results suggested that PL could modify plasma lipoprotein metabolism and lipid profile, and that the optimal dietary PL level may well exceed 80 g kg⁻¹ for cobia larvae according to growth and survival.     

Dietary n‐3 LC‐PUFAs affect lipoprotein lipase (LPL) and fatty acid synthase (FAS) activities and mRNA expression during vitellogenesis and ovarian fatty acid composition of female silver pomfret (Pampus argenteus) broodstock

We studied the effects of dietary n‐3 LC‐PUFAs on the activities and mRNA expression levels of tissue lipoprotein lipase (LPL) and fatty acid synthase (FAS) during vitellogenesis and ovarian fatty acid composition in female silver pomfret broodstock. Broodstock were fed one of four experimental diets for 185 days: FO (100% fish oil), FSO (70% fish oil + 30% soybean oil), SFO (30% fish oil + 70% soybean oil) or SO (100% soybean oil). The results revealed that hepatic LPL and FAS and ovarian FAS activities and mRNA expression levels significantly increased at vitellogenesis and postvitellogenesis relative to previtellogenesis, with no significant differences between these two stages, except for hepatic LPL mRNA expression. Dietary n‐3 LC‐PUFAs decreased tissue FAS and increased LPL activities and mRNA expression levels. The ovarian concentrations of 20:4n‐6 (ARA), 20:5n‐3 (EPA), 22:6n‐3 (DHA) and n‐3 LC‐PUFAs were directly influenced by n‐3 LC‐PUFA levels. Total n‐3 LC‐PUFA concentrations in SO were 57% lower than those in FO, while 18:2n‐6 concentrations in SO were 4.7 ×  higher than those in FO. These results revealed that high dietary n‐3 LC‐PUFAs levels significantly affected tissue lipid metabolism in female silver pomfret broodstock during vitellogenesis by upregulating LPL and downregulating FAS.     

cDNA cloning and expression of a novel group IB phospholipase A2 in the intestine of the red sea bream,Pagrus (Chrysophrys) major

A cDNA encoding a novel phospholipase A₂ (PLA₂), which was named IN PLA₂, was cloned from the intestine of the red sea bream. The amino acid sequence of IN PLA₂ showed 49–75% homology with those of red sea bream group IB sPLA₂, hepatopancreas DE-1 and DE-2 PLA₂, and gill G-3 PLA₂. IN PLA₂ consists of a prepropeptide of 24 amino acid residues, followed by a mature protein. IN PLA₂ contains 14 cysteines, and includes Cys11, the calcium binding loop and the pancreatic loop that are commonly conserved in group IB sPLA₂ enzymes. In addition, IN PLA₂ is a cationic protein with a pI of 8.52. Therefore, IN PLA₂ was identified as a novel group IB sPLA₂ isoform in red sea bream. IN PLA₂ mRNA was found by northern blot analysis to be expressed mainly in the pyloric caeca and the intestine, and was detected in the goblet cells of the intestine by in situ hybridization. The expression level of IN PLA₂ mRNA was elevated by intravenous injection of lipopolysaccharide—the outer-membrane component of Gram-negative bacteria. These results suggest that IN PLA₂ is secreted from the goblet cells of the intestine in response to stimulus such as bacterial infection, and that it contributes to antimicrobial defense in addition to the digestion of dietary phospholipids in the gastrointestinal tract.     

Protein‐sparing effects and LPL gene expressions of dietary lipids in the juvenile soft‐shelled turtle,Pelodiscussinensis

The aim of this study was to evaluate the effects of dietary lipids on protein‐sparing and lipoprotein lipase (LPL) mRNA expression in culture using 360 juvenile soft‐shelled turtles (Pelodiscussinensis) (initial weight 4.26 ± 0.14 g). The turtles were allotted to six diets with three duplicates for 60 days. A control diet with 46% protein and 55% fishmeal (CD) and five isonitrogenous diets with 41.3% protein and 45% fishmeal (F, S, L1, L2 and L3) were used, containing the following three lipid types: fish oil, soybean oil and mixed oils (soybean oil: fish oil = 1:1). The results showed that the survival rate was not affected by dietary lipids (P > 0.05). The highest weight gain and lowest feed coefficient ratio were seen in the L3 diets (P < 0.05). Turtles fed with L2 and L3 diets had lower superoxide dismutase activities, higher alanine aminotransferase activities and higher cholesterol concentrations than those exposed to other diets (P < 0.05). Hepatic LPL activity and LPL mRNA expression were higher in the L3 diets than in the other diets (P < 0.05). Overall, there were obvious protein‐sparing effects of dietary lipids and LPL mRNA expression was stimulated by high dietary lipids in soft‐shelled turtles in this study.