The effects of vitamin E on non-specific immune response of the juvenile soft-shelled turtle <Emphasis Type="Italic">Pelodiscus sinensis</Emphasis>

ABSTRACT:   This study was conducted to determine the effect of dietary vitamin E supplements on non-specific immune responses in juvenile soft-shelled turtles. Turtles were fed diets supplemented with vitamin E at 0 (control), 50, 250, 500, 1000 and 5000 mg/kg, respectively, for 4 weeks. Results showed that blood cell phagocytosis and serum bactericidal activity were significantly improved in turtles from two diets (250 and 500 mg/kg vitamin E supplementation), while had no significant improvement in turtles from the three remaining diets (50, 1000 and 5000 mg/kg vitamin E supplementation) when compared to turtles from the control group. Serum bacteriolytic activity in turtles from diets with 50, 250, 500 and 1000 mg/kg vitamin E supplementation were higher than activity levels quantified for the control group, but no significant difference was observed between the 5000 mg/kg vitamin E supplementation diet and the control diet. The results suggest that vitamin E has an upper and lower threshold for improving non-specific immune function in soft-shelled turtles, and the optimal supplementation may be between 250 and 500 mg/kg.     

Effects of dietary vitamin E level on growth and tissue lipid peroxidation of soft-shelled turtle, Pelodiscus sinensis (Wiegmann)

A feeding trial was conducted to determine the effect of dietary vitamin E supplementation on growth, liver lipid peroxidation and liver and muscle vitamin E level of soft‐shelled turtle, Pelodiscus sinensis. Eight experimental diets analysed to contain 0–457 IU vitamin E kg^?1 were fed to juvenile soft‐shelled turtle of 4.8 g initial body weight for 12 weeks. Weight gain (WG) of the turtles fed the diet containing no vitamin E was significantly lower than those fed diets containing 83–457 IU vitamin E kg^?1 (P<0.05). Feed conversion ratio and protein efficiency ratio showed similar trends to that of WG. No significant difference (P>0.05) was found in whole‐body composition among turtles fed the different diets. Dietary vitamin E requirement using WG as the response and estimated using the broken‐line regression model is approximately 88 IU kg^?1. Liver and muscle vitamin E content increased when dietary vitamin E level increased. Ascorbate‐induced lipid peroxidation in liver tissue of turtles fed diets containing 0 and 17 IU vitamin E kg^?1 was significantly (P<0.05) greater than those fed diets containing high vitamin E (≥35 IU kg^?1).     

中华鳖对嗜水气单胞菌粗脂多糖(LPS)的免疫应答

将采用温酚法从嗜水气单胞菌(Aeromonas hydrophila)、柱状嗜纤维菌(Cytophaga columnaris)和叉尾鮰爱德华菌(Edwardsiella ictaluri)中提取的粗脂多糖(crude lipopolysaccharide,LPS)作为免疫原,注射接种中华鳖(Pelodiscus sinensis)后,通过测定受免鳖的交叉凝集抗体效价、肾脏和血液中吞噬细胞的吞噬活性和对A.hydrophila活菌细胞内杀菌活性,比较了3种致病菌的粗LPS对中华鳖的免疫原性。试验结果表明,从3种致病菌中提取的粗LPS对中华鳖均具有较强的免疫原性,受免鳖的血清中存在对3种致病菌的交叉凝集抗体,与对照鳖相比,肾脏和血液中吞噬细胞对吞噬原的吞噬活性和对A.hydrophila活菌杀菌活性明显上升。说明在3种供试菌的粗LPS上不同程度地存在3种致病菌的共同保护性抗原。     

复方中草药对中华鳖营养免疫及抗氧化功能的影响

为研究复方中草药对中华鳖营养免疫及抗氧化功能的影响,试验在水温27~29℃时,将体质量25~35g的中华鳖(Pelodiscus sinensis)幼鳖饲养在池底铺10cm细沙的1.0m×0.8m×0.8m水泥池中,分别投喂添加0%(对照组)、1%、3%和5%复方中草药(芹菜籽粉∶葡萄籽粉∶薏仁粉=1:1:1)的饲料。养殖60d后主要测定幼鳖血液中的抗氧化指标。结果表明:1%、3%、5%中草药组血清谷胱甘肽活性(GSH-Px)均显著高于对照组,剂量效应规律较为明显。5%中草药组血清总抗氧化能力(T-AOC)显著高于对照组。1%中草药组血清过氧化氢酶(CAT)活性显著高于对照组,剂量效应规律不明显。1%、3%和5%中草药组血清丙二醛(MDA)含量均显著低于对照组。3%和5%中草药组幼鳖血清尿素氮(BUN)水平显著低于对照组,表明中草药可提高中华鳖幼鳖蛋白质利用效率。综上所述,从中华鳖幼鳖血清T-AOC活性、MDA含量以及BUN水平等指标来看,结合经济性原则,该复方中草药添加量以3%较为适宜。另外,该复方中草药对于中华鳖幼鳖整个机体的免疫状态的影响规律不明显,其改善中华鳖抵抗力的内在机制还有待深入研究。     

Effects of dietary starch levels on the growth,plasma metabolites and expression of genes involved in glucose transport and metabolism in Pelodiscus sinensis juveniles

A 10‐week feeding trial was conducted to evaluate the growth performance, glucose transport and metabolism of Chinese soft‐shelled turtles (Pelodiscus sinensis) exposure to graded levels of dietary starch (0.52%, 7.43%, 14.74%, 22.99% and 31.38%). The 360 turtles (initial body weight, 12.94 ± 0.50 g) with 12 replicates were randomly assigned to five experimental diets. The highest weight gain and specific growth rate (SGR) were observed in 7.43% group and the lowest in 31.38% group. The protein efficiency ratio, whole‐body lipid contents, hepatic glycogen contents and the 4‐hr postprandial plasma glucose levels were significantly increased with the increment of starch levels (p < .05). In contrast, the daily feed intake and feed conversion ration were significantly declined (p < .05). The mRNA levels of glucose transporter 2, glucokinase, pyruvate kinase, malic enzyme and acetyl‐CoA carboxylase alpha genes in the liver significantly increased as the increase in starch levels at 4‐hr and 24‐hr post feeding (p < .05). No significant differences were observed in the expression of gluconeogenesis genes at each time point (p > .05). These results suggested that dietary addition of starch up‐regulated hepatic glycolysis, glycogenesis and lipogenesis genes expression, but the deficient response of gluconeogenesis to dietary starch might be part of the causes limited the starch utilization. Based on the secondary polynomial regression of SGR, y = ?0.0011x² + 0.028x + 1.63 (R² = 0.9292), the 12.73% inclusion level of dietary starch was recommended in juvenile turtles.     

中华鳖GnRH 1基因cDNA克隆及表达特征

为了解GnRH基因在中华鳖(Pelodiscus sinensis)性腺和胚胎发育过程中的表达特征,采用cDNA末端快速扩增(RACE)技术从中华鳖全脑中获得与生长生殖调控密切相关的GnRH1基因全长cDNA,并运用实时荧光定量PCR(qRT-PCR)技术检测GnRH1在成鳖不同组织和胚胎发育时期的表达水平。结果显示:中华鳖GnRH1基因cDNA全长546 bp,其中5′非编码区(5′UTR)99 bp,3′非编码区(3′UTR)168 bp,开放阅读框(ORF)279 bp,编码92个氨基酸,分子质量为10.23 ku,理论等电点pI为5.65,具有N端信号肽(1~23 aa)、核心十肽区域(24~33 aa)、断裂位点GKR(34~36 aa)及相关肽区域(37~92 aa),符合GnRH蛋白典型结构特征。系统进化树结果显示,中华鳖GnRH1基因和绿海龟(Chelonia mydas)、墨西哥箱龟(Terrapene carolina mexicana)及西部锦龟(Chrysemys picta bellii)GnRH1基因聚为一支。qRT-PCR结果表明,GnRH1基因在中华鳖雌雄个体的8个组织中均有表达,在脑和性腺组织中高表达,且具有性别差异,雄性中华鳖中的表达显著高于雌性(P<0.05);在10个胚胎发育时期均表达,且随发育时间的后移,表达量显著增加,在第16期达到峰值。GnRH1基因可能在中华鳖生长及性腺分化中具有重要作用。     

中华鳖日本品系对6种饲料蛋白原料表观消化率的研究

研究了中华鳖日本品系(Pelodiscus sinensis,Japanese strain)幼鳖对蝇蛆蛋白粉、虾壳粉、发酵豆粕、豌豆浓缩蛋白、大豆浓缩蛋白以及玉米蛋白粉的干物质、粗蛋白质、氨基酸、粗脂肪和总能的表观消化率。试验饲料由70%基础饲料和30%待测饲料原料组成,并以0.5%的三氧化二铬(Cr2O3)为外源指示剂。选取平均体重为(14.69±0.55)g的中华鳖日本品系幼鳖180只,随机分成6组,每组3个重复,每个重复10只,投喂试验饲料养殖4周,采用虹吸法收集粪便进行表观消化率测定。结果表明,6组饲料的干物质、粗蛋白质、粗脂肪以及总能表观消化率范围为77.8%~85.6%、73.2%~83.7%、81.2%~91.1%和78.8%~85.6%,其中蝇蛆蛋白粉和发酵豆粕的干物质表观消化率较高,大豆浓缩蛋白的粗蛋白、粗脂肪及总能表观消化率较高;各原料总氨基酸表观消化率在75.8%~86.1%,最高值和最低值分别出现在大豆浓缩蛋白和虾壳粉。     

不同雌雄配比对中华鳖人工繁殖受精率的影响

研究了中华鳖(Pelodiscus sinensis)人工繁殖不同雌雄配比的受精率差异,结果显示:在放养密度3750~4500 kg/hm2,亲鳖规格♀1.5 kg/只、♂2.0 kg/只条件下,雌雄配比可达8∶1。在不增加水体面积和投资情况下,雌雄比8∶1与5∶1相比多产受精卵13500~18000枚/hm2,可增加纯利润27000~30000元/hm2。