Effects of Dietary Protein Level on Nitrogen and Energy Budget of Juvenile Chinese Soft‐shelled Turtle,Pelodiscus sinensis,Wiegmann

The effects of dietary protein level on nutrient digestibility and utilization of protein and energy of juvenile Chinese soft‐shelled turtle, Pelodiscus sinensis, were investigated. Seven isoenergetic diets containing graded levels of protein (25, 27, 29, 31, 33, 35, and 43%) were fed to turtles initially averaging 4.8 ± 0.7 g for 8 wk at 30 ± 0.5 C. The apparent digestibilities of dry matter, protein, and energy were generally increased with dietary protein level. Proportions of nitrogen intake (CN) allocated to excretory nitrogen (UN) were higher in 25 and 43% protein diets than the other diets. UN proportion was lowest in the 33% protein diet, whereas retained (growth) nitrogen (RN) followed an inverse pattern with UN. Percentages of gross energy intake (IE) used for excretory energy (UE) generally increased when dietary protein levels increased. Turtles fed 25–29% protein diets showed a markedly higher proportion of metabolizable energy (ME) than those fed 31–43% protein diets. Analysis by second‐order polynomial regression using RN as the indicator indicated that the estimated optimal dietary protein level was 34.6%. The lowest and highest percentages of retained (growth) energy (RE) were observed in the 25 and 33% protein groups. Nitrogen and energy budget equations of turtles fed 33% protein diet were 100 CN = 8.22 FN + 37.48 UN + 54.30 RN and 100 IE = 10.88 FE + 2.82 UE + 65.60 ME + 20.69 RE, respectively. Diets with excessive protein resulted in high proportions of protein and energy used for excretion.     

Thiamine requirement of Chinese soft‐shelled turtle,Pelodiscus sinensis (Wiegmann)

The objective of the present study was to estimate the dietary thiamine (vitamin B₁) requirement of juvenile soft‐shelled turtles, Pelodiscus sinensis. Eight experimental diets containing 0, 1.90, 3.63, 5.65, 7.51, 9.62, 11.37 and 13.64 mg thiamine/kg diet were fed to 160 soft‐shelled turtles reared individually for 10 weeks. The turtles had an average weight of 5.33 ± 0.21 g. Among all the dietary groups, weight gain, feed utilization and tissue thiamine were the lowest in the turtles fed with a thiamine‐free diet. The variables increased when dietary thiamine increased and then levelled off beyond 3.63 mg/kg diet. In contrast, plasma pyruvate and lactate concentrations in turtles decreased when dietary thiamine increased and then levelled off beyond 3.63 and 5.65 mg/kg diet respectively. Estimation using a broken‐line regression model, the vitamin B₁ requirement of soft‐shelled turtles was 3.2 mg thiamine/kg diet based on weight gain and plasma lactate concentration. When tissue thiamine and plasma pyruvate concentrations were used as the indicators, the estimated requirement was 5.4–6.5 mg/kg diet.     

Estimation of dietary vitamin A requirement of juvenile soft‐shelled turtle,Pelodiscus sinensis

A feeding trial was conducted to evaluate the dietary vitamin A requirement of the soft‐shelled turtle Pelodiscus sinensis. Turtles with an approximate body weight of 6.8 g were fed diets containing 0–4.58 mg kg^?1 (15 251 IU) vitamin A (retinyl acetate) for 8 weeks. Turtles fed diets without vitamin A showed the lowest growth performance. The animals gradually gained weight when dietary vitamin A increased from 0 to 3.6 mg kg^?1. Liver vitamin A gradually increased with supplementation of dietary vitamin A of up to 2.7 mg kg^?1. Beyond this concentration, the vitamin A in the liver significantly increased. Data analysis using a polynomial regression or exponential model showed that the estimated dietary vitamin A requirement for juvenile soft‐shelled turtles based on weight gain, red blood cell count and liver vitamin A measurements was 3.48, 2.84 and 2.58 mg kg^?1, respectively.     

Effects of Taurine Supplementation on Growth Performance and Antioxidative Capacity of Chinese Soft‐shelled Turtles,Pelodiscus sinensis,Fed a Diet of Low Fish Meal Content

This experiment examined the effects of taurine supplementation on the growth performance and antioxidative capacity of Chinese soft‐shelled turtle, Pelodiscus sinensis, fed diets of low fish meal content. The base diet (with 0.31% taurine) consisted of low fish meal level and had an animal : plant protein ratio of 2:1. Six experimental diets were supplemented with various taurine levels (0, 0.25, 0.5, 0.75, 1.0, and 1.25%) for 8 wk. There were significant quadratic regression relationships between the taurine supplementation level and specific growth rate (SGR), feed conversion ratio (FCR), and protein efficiency ratio (PER). All taruine supplemented groups showed higher SGR and PER and lower FCR than the group fed diet without taurine supplementation. Significant quadratic regressions were also showed between the supplemented taurine level and the total superoxide dismutase activity, malondialdehyde content, and total antioxidant capacity. The results indicate that taurine supplementation in a diet with low fish meal level can improve the growth of turtles. With comprehensive analysis on the effects of taurine supplementation on the growth performance and antioxidative capacity of turtles, the optimal taurine supplementation level was suggested to be 0.9% in the diet with an animal : plant protein ratio of 2:1.     

Estimation of dietary vitamin K requirement of juvenile Chinese soft‐shelled turtle,Pelodiscus sinensis

A 10‐week feeding trial was conducted to estimate the dietary vitamin K requirements of juvenile Chinese soft‐shelled turtles, Pelodiscus sinensis, using menadione sodium bisulphite (MSB) as the vitamin K source. Juvenile soft‐shelled turtles with 6.1 ± 0.1 g body weight were individually reared and fed diets containing seven levels of MSB (0.0, 5.0, 8.9, 12.3, 16.6, 20.7 and 41.0 mg/kg). Turtles fed with an MSB‐free diet exhibited the lowest feed utilization, carapace Ca concentrations, carapace strength or liver menaquinone (MK‐4) concentrations among all dietary groups. Weight gain (WG) in the turtles tended to increase with increase in dietary MSB until a dietary MSB level of 16.6 mg/kg, where WG levelled off. Total plasma prothrombin concentrations, an indicator for blood clotting function, in the turtles fed on MSB supplemented diets were significantly (p < 0.05) lower than those in turtles fed on MSB supplemented diets. Liver MSB concentrations increased with increase in dietary MSB. Using broken‐line or sigmoidal regression model, the vitamin K requirements of juvenile Chinese soft‐shelled turtles based on WG, total plasma prothrombin concentrations and liver vitamin K were 21.5, 25.8 and 29.9 mg MSB per kg, respectively.     

鸡肝脏和配合饲料对稚鳖生长的影响

采用人工配合饲料和鸡肝脏,饲喂体重２．８４￣５．２２ｇ的稚鳖３０ｄ。鸡肝脏饲养的稚鳖增重和增宽均及显著地高于高售饲料;鸡肝脏的饲料系数仅为市售饲料的５４．５％。鸡肝脏饲喂后鳖的能值、蛋白质含量都有所提高,脂肪含量变化不大,而灰分和钙、镁含量下降;市售饲料饲养后,能值、灰分和钙、镁含量都有提高,蛋白质和肥肪含量下降。     

Efficacy and tolerance of lutein as colourant in diet of juvenile soft‐shelled turtle Pelodiscus sinensis

The effects of dietary supplementation of lutein on the growth, haematological profile and pigmentation in target tissues of the soft‐shelled turtles were investigated. Five experimental diets were prepared with lutein levels at 1.16 (D1, control), 70.3 (D2), 132 (D3) and 239 (D4) mg kg^?1, and one more high inclusion level of 3410 mg kg^?1 (D5) was included for tolerance test. Each diet was fed to 24 juvenile turtles for 8w. Specific growth rate (SGR) in D2, D3 and D4 was significantly higher than that of group D1 and D5. Skin yellowness (b*) and chroma (C*) of both carapace and plastron skin were all significantly increased with higher dietary lutein levels. Dietary lutein supplementation turned the H° values of skins into yellowness. The accumulation of lutein in skin and muscle also augmented significantly with increasing lutein levels and fit a logarithmic correlation with dietary lutein levels. There were almost no differences in haematological parameters of all turtles. Lutein is an efficient and safe colourant in diet of soft‐shelled turtle. The optimal inclusion levels for the highest SGR and desired skin colour in carapace and plastron of soft‐shelled turtle were 162, 142 and 143 mg kg^?1, respectively.     

Newly identified C‐type lysozyme in Chinese soft‐shelled turtle (Pelodiscus sinensis) exhibits potent antimicrobial activity

Lysozymes play vital roles in humoural immune response against bacterial invasion by its lytic activity. In the present study, a new C‐type lysozyme was identified and characterized from Chinese soft‐shelled turtle Pelodiscus sinensis. The full‐length cDNA of PslysC was of 923 bp, encoding a polypeptide of 148 amino acid residues. The multiple alignments and phylogenetic relationship analysis revealed the highly enzyme‐related conserved residues. The real‐time PCR analysis suggested that PslysC was constitutively expressed in a wide range of tissues with highest level in blood cells and liver. The expression of PslysC could be significantly up‐regulated under Aeromonas jandaei infection and ammonia exposure, while no significant changes were found under Poly I:C infection. The rPslysC protein was expressed in E. coli and purified by Ni‐NTA. The optimal pH and temperature for rPslysC protein lytic activities were determined at pH 7 and 30℃. rPslysC can inhibit the growth of eight kinds of Gram‐negative bacteria, and three kinds of Gram‐positive bacteria. The binding activity of rPslysC to different microbial polysaccharides and microorganism was analysed. The results showed that rPslysC could bind to selected bacteria, and exhibit a strong binding activity to lipopolysaccharide and peptidoglycan, but a weak binding activity to β‐glucan. This suggests that the binding activity might be the major mechanism of action to realize the antibacterial activity. The present study will provide helpful evidence to further understand the innate immunity of P. sinensis, and the interaction mechanisms of C‐type lysozymes with bacterial membranes.     

Effects of dietary zinc levels on growth,serum zinc,haematological parameters and tissue trace elements of soft‐shelled turtles,Pelodiscus sinensis

A 10‐week feeding trial was conducted to evaluate the effects of dietary zinc (Zn) contents on the growth, tissue trace element contents and serum Zn levels in soft‐shelled turtles, Pelodiscus sinensis. Juvenile soft‐shelled turtles approximately 4.8 g in body weight were fed casein‐based diets containing seven levels of Zn (14, 23, 32, 43, 58, 87 and 100 mg kg^?1) for 10 weeks. There were no significant differences (P > 0.05) in weight gain (WG), feed conversion ratio (FCR) or protein efficiency ratio (PER) among the dietary treatments. However, Zn concentrations in the liver, serum and carapace of turtles fed the basal diet containing 14 mg Zn kg^?1 were the lowest among all groups. Zn contents in the liver, serum and carapace increased when dietary Zn increased up to a dietary Zn level of approximately 43 mg kg^?1. Beyond this dietary level, tissue Zn contents were relatively constant. Carapace iron (Fe), selenium (Se) in hard tissues and haemoglobin concentrations decreased when dietary Zn increased. Dietary Zn requirements of juvenile soft‐shelled turtles derived from regression modelling using the liver, serum, carapace and bone Zn contents as indicators were 42, 39, 35 and 46 mg Zn kg^?1, respectively.     

Effects of dietary magnesium on the growth,carapace strength and tissue magnesium concentrations of soft‐shelled turtle,Pelodiscus sinensis (Wiegmann)

A feeding trial was conducted to evaluate the effects of dietary magnesium on the growth, carapace strength, tissue and serum Mg concentration of soft‐shelled turtles, Pelodiscus sinensis (Wiegmann). Juvenile soft‐shelled turtles of approximate 5.4 g body weight were fed diets with seven levels of Mg (48, 206, 369, 670, 955, 1195 and 1500 mg Mg kg^?1) for eight weeks. No significant difference (P ≥ 0.05) was found in weight gain (WG), feed conversion ratio or protein efficiency ratio among treatments. However, the WG of turtles continued to increase with increasing dietary Mg levels up to 670 mg kg^?1, beyond which the WG levelled off. The plasma alkaline phosphatase activity and the muscle, bone Mg concentrations of the turtles increased with the increasing dietary Mg levels between 48 and 955 mg kg^?1, beyond which the tissue Mg concentrations remained relatively constant. Furthermore, the carapace strengths of turtles fed with the control diet of 48 mg Mg kg^?1 were significantly weaker (P < 0.05) than that of turtles fed with diets containing higher Mg levels. Based on a broken‐line modelling analysis, the required dietary Mg level for the optimal WG of juvenile soft‐shelled turtles was estimated to be approximately 650 mg kg^?1. By contrast, the required dietary Mg levels for turtles to reach the optimal muscle and bone Mg concentrations were 1050 and 1000 mg kg^?1 respectively. The required dietary Mg level for maximal alkaline phosphatase activity was approximately 980 mg kg^?1.     

Effects of fishmeal replacement with cottonseed meal protein hydrolysate on growth,digestion and intestinal histology of juvenile Chinese soft‐shelled turtle,Pelodiscus sinensis

A 60‐day feeding experiment was conducted to evaluate the effects of fishmeal (FM) replacement with cottonseed meal protein hydrolysate (CPH) on growth, digestion and intestinal histology of juvenile Chinese soft‐shelled turtle (Pelodiscus sinensis). Five diets were formulated to replace 0, 50, 100, and 150 g/kg fishmeal protein by CPH (CPH0, CPH5, CPH10, CPH15) and CPH15L (CPH15 with micro capsule‐L‐lysine). Weight gain, feed conversion rate and protein efficiency ratio showed no significant differences compared to control group (p > .05). The highest feed intake indicated in CPH15 (p < .05). The composition of whole‐body varied slightly in each groups (p > .05). The trypsin activity significantly elevated when dietary fishmeal protein was replaced by CPH at 30–90 g/kg (p < .05). A significantly higher lipase activities in CPH5 than control group (p < .05). The CPH5‐10 groups showed higher villus height than the other groups (p < .05). The microvillus length in turtles with CPH showed a significant increasing length (p < .05). The results indicated that replacing up to 90 g/kg of dietary fishmeal protein with CPH did not hamper growth or reduce feed intake of turtles. Moreover, CPH replaced 60 g/kg FMP can increase intestine digestive enzymes activities and improve intestinal development.     

Acute and Chronic Effects of Ammonia on Juvenile Cuttlefish,Sepia pharaonis

The aim of this study was to provide a reference value for the safe regulation and control of ammonia nitrogen in the aquaculture of Sepia pharaonis. The effects of acute and chronic toxicity of ammonia on the cuttlefish, S. pharaonis, were tested experimentally using juvenile S. pharaonis. The results showed that the half‐lethal concentration (LC₅₀ ) values of ammonia nitrogen in juvenile S. pharaonis with a body weight of 6.52 ± 0. 23 g at 24, 48, 72, and 96 h were 31.72, 25.77, 23.33, and 18.33 mg/L, respectively, and the corresponding un‐ionized ammonia nitrogen (UIA‐N) concentrations were 1.66, 1.35, 1.22, and 0.96 mg/L, respectively. Compared with the control, the survival rate, specific growth rate, and feed intake of juvenile S. pharaonis declined significantly, and the feed conversion ratio and hepatosomatic index increased significantly at 56 d after exposure to >1 mg/L ammonia nitrogen. Juvenile S. pharaonis should be maintained at a concentration of ammonia nitrogen of no more than 1 mg/L (UIA‐N is 0.056 mg/L) in culture, and removing harmful nitrogenous wastes from the seawater is critical in maintaining cuttlefish culture.     

Effects of dietary β‐carotene levels on growth and liver vitamin A concentrations of the soft‐shelled turtle,Pelodiscus sinensis (Wiegmann)

A feeding trial was conducted to evaluate the effect of dietary β‐carotene level on the growth and liver vitamin A concentrations in soft‐shelled turtles, Pelodiscus sinensis, fed a vitamin A‐free diet. Soft‐shelled turtles were fed diets containing 0, 14.5, 26.5, 47.5, 87.3, 112.8 and 163.8 mg β‐carotene kg^?1 for 10 weeks. Although it was not statistically significant due to high deviation within each group, mean weight gain of soft‐shelled turtles fed the diet without β‐carotene supplementation was the lowest among all test groups. Vitamin A concentrations in liver of turtles significantly (P<0.05) increased when dietary β‐carotene level reached 47.5 mg kg^?1 indicating that soft‐shelled turtles were capable of converting β‐carotene to vitamin A. Analysed by regression modelling, dietary β‐carotene levels for optimal growth and maximal liver vitamin A contents of juvenile soft‐shelled turtles fed the vitamin A‐free diets were 49.1 and 88.7 mg kg^?1 respectively.     

Different preference is modulated by the feeding stimulants supplementation in different Chinese soft‐shelled turtle (Pelodiscus sinensis) basic diets

Five 2‐week feeding trials were conducted to investigate five stimulants on P. sinensis. Two isonitrogenous and isoenergetic diets were formulated, one contained 600 g kg^–1 fishmeal (FM diet) and the other contained 420 g kg^–1 fishmeal and 225 g kg^–1 animal protein blend (APB diet), and each tested one stimulant with four levels. The feeding stimulant candidates were betaine, 2‐carboxy‐ethyl dimethyl sulphonium bromide (DMPT), inosine‐5′‐monophosphate (IMP), taurine and squid extract. In the experiment on each stimulant, turtles (10.85 ± 0.10 g) were equally divided into FM and APB groups and fed with an equal mixture of corresponding diets containing four levels of stimulants, respectively. Each diet contained a unique rare earth oxide as inert marker. Turtles were fed twice daily (8:00 and 17:00), and faeces were collected. Preference for each diet was estimated based on the relative concentration of each marker in the faeces. In the FM group, only IMP showed the enhanced attraction. But in APB groups, all the stimulants, except IMP, showed higher preference than basal diets for at least one inclusion level, and the optimum level was 10 g kg^–1 for betaine, 0.1 g kg^–1 for DMPT, 0.1 g kg^–1 for IMP, 5 g kg^–1 for taurine and 10 g kg^–1 for squid extract, and the squid extract had the strongest stimulating effect among the stimulants.     

Partial replacement of fish meal by soy protein concentrates in diets for a new Japanese strain of juvenile soft‐shelled turtle,Pelodiscus sinensis

An 8‐week experiment was conducted to evaluate the effects of dietary fish meal (FM) replaced by soybean protein concentrate (SPC) on Japanese strain of soft‐shelled turtle, Pelodiscus sinensis juveniles. Diets were formulated to replace FM protein by SPC at 0, 15, 30, 45, 60 or 60% supplemented with phytase (2000 FTU kg^?1) (designated as S0, S15, S30, S45, S60 and S60P, respectively), and each diet was fed to triplicate groups. The results showed that the growth was significantly lower when dietary SPC replaced more than 45% FM. The turtles fed the S15 or S30 diet showed comparable feed and protein utilization efficiency compared with the S0 group, whereas more than 30% replacement of FM adversely affected these values. Increasing dietary SPC levels significantly lowered the apparent digestibility coefficients (ADC) of dry matter, protein, lipid, phosphorus and gross energy. Whole‐body protein, ash and phosphorus content showed a declining trend when dietary SPC levels increased, while body lipid and moisture content were unaffected. When the turtles were fed diets with increasing levels of SPC, serum total protein concentration, alkaline phosphatase and catalase activities decreased with a corresponding increase in glutamic pyruvic transaminase activity. Turtles fed the S60P diet showed comparable growth performance and feed utilization efficiency to the S40 group, and were superior to the S60 group. The present study showed that SPC could successfully substitute for 30% FM protein in the diets for P. sinensis juveniles, and the maximum effective substitution may be greater if exogenous phytase was added.     

Effects of fish meal replacement with animal protein blend on growth performance,nutrient digestibility and body composition of juvenile Chinese soft‐shelled turtle Pelodiscus sinensis

An 8‐week feeding trial was conducted to evaluate the potential of partial replacement of fish meal with rendered animal protein blend (APB) [meat and bone meal (MBM): expanding blood meal (EBM) = 4 : 1] in the diet of juvenile Chinese soft‐shelled turtle Pelodiscus sinensis. Seven isonitrogenous and isoenergetic practical diets replacing 0%, 10%, 20%, 30%, 40%, 50% and 60% fish meal protein by APB were formulated. Weight gain, feed efficiency, protein efficiency and the apparent digestibility coefficients (ADCs) of dry matter and gross energy in the experimental diets all exhibited no significant difference (P > 0.05) when dietary fish meal was replaced for 0–40%, but decreased significantly (P < 0.05) with further increasing substitution levels. However, relative feed intake, ADC of lipid as well as the contents of whole‐body moisture, protein, lipid and ash all showed little differences (P > 0.05) among all the treatments. Nitrogen retention and the ADCs of protein and most amino acids all decreased significantly (P < 0.05) when fish meal was substituted by more than 30%. The results indicated that fish meal in the practical diet of juvenile Chinese soft‐shelled turtle could be replaced by 30–40% using a combination of MBM and EBM without negative effects on growth performance, nutrients digestibility and body composition.     

果寡糖和糖萜素对中华鳖生长性能及免疫功能的影响

选用2龄平均体重150g左右的中华幼鳖54只，按2因素4水平的正交试验设计，随机分成9组，每组各接受1种处理。第9处理组（FOS=1000mg/kg，SHP=1000mg/kg）日增重量大（1.75g），第1处理组（FOS=200mg/kg，SHP=200mg/kg）最小；第9处理组饲料系数最低（1.80），第1处理组最高（2.13）。第8处理组（FOS=1000mg/kg,SHP=600mg/kg）胸腺最重（0.26g），第1处理组最低（0.21g）。脾脏的变化趋势与胸腺相似。第8处理组胸腺指数最大（1.13），第1处理组最低（0.98），而脾脏指数变化趋势与胸腺指数相似，果寡糖和糖萜素适宜添加量为600-1000mg/kg。     

甲鱼最佳养殖温度的探讨

采用适量投喂法对稚鳖对29.5℃，30℃，30．5℃，31℃，32℃等不同温度下的摄食及生长进行研究，发现在30．7℃稚鳖个体增重最快，29．8℃饲料系数最低，在30℃以内，稚鳖的生长速度随着水温的上升而加快，当水温超过30℃时，稚鳖的生工速度却随着水温的上升而下降，在不同的湿度下，稚鳖的生长速度差异显著，日投饲率为2．8％－3．0％，全天投喂的饲料中，上午占1／3，下午或晚占2／3。     

几种促生长剂对中华鳖养殖效果的研究

研究了免疫促生长剂C、黄霉素和维吉尼霉素对中华鳖稚鳖生长及饲料转化效率的影响。结果表明。免疫促生长剂C具有明显促生长作用。生长比速高于对照组14．2％。免疫促生长剂C、黄霉素和维吉尼霉素均明显提高了蛋白效率。分别高于对照组39．9％、36．8％和29．34％。而饲料系数明显降低。分别较对照组降低28．6％、27．0％和22．6％。维吉尼霉素、黄霉素明显提高了肝体指数，分别高于对照组27．5％和21．9％。3种促生长剂均具有明显促进营养物质利用的作用。尤以免疫促生长剂C作用最为显著。均可考虑在中华鳖饲料中使用。     

Effects of Salinity on Growth and Energy Budget of Juvenile Cobia,Rachycentron canadum

The effects of salinity on the growth and energy budget of juvenile cobia, Rachycentron canadum, were evaluated. Triplicate tanks with ten fish per tank (initial weight 17.58 ± 0.26 g/fish, mean ± SD) reared at salinities of 5, 10, 15, 20, 25, 30, and 35 ppt were fed with fresh squid to satiety for 15 d. Results indicated that there were no significant differences in daily ration level in wet weight (RL_w), dry weight (RL_d), and energy (RL_e) of the fish. There were also no significant variations in daily fecal production (f_e) and apparent digestibility coefficient of energy (ADC_e) among salinity treatments. Specific growth rates (SGRs) in wet weight (SGR_w), dry weight (SGR_d), and energy (SGR_e) showed domed curves relative to salinity. Quadratic regression analyses of SGR_w, SGR_d, and SGR_e against salinity indicated that the optimal salinity for maximal growth of juvenile cobia was 29.9, 29.9, and 28.5 ppt, respectively. Similar to the trend of SGR, food conversion efficiency for juvenile cobia in wet weight (FCE_w), dry weight (FCE_d), and energy (FCE_e) increased with the increases in salinity, maximized at 30 ppt, and then decreased when salinity reached 35 ppt.