White-spot disease of Chinese soft-shelled turtles (Trionyx sinens) caused by Paecilomyces lilacinus

Chinese soft-shelled turtles （Trionyx sinens） in culture farms using an artificial warming system in Zhejiang, China, often show typical signs of white-spot disease such as white spots on their bodies, skin lesions, anorexia and eventually death. The sick turtles were mostly 5-80 g in weight. A suspected fungal pathogen was isolated from the sick turtles and verified as Paecilomyces lilacinus by sequence analysis of the internal transcribed spacer （ITS） of its ribosomal DNA （rDNA）. Detailed morphological examinations were also conducted to confirm the white-spot disease.

Effects of Brown Fish Meal Replacement with Fermented Soybean Meal on Growth Performance, Feed Efficiency and Enzyme Activities of Chinese Soft-shelled Turtle, Pelodiscus sinensis

A 120-day feeding experiment was conducted to investigate the effects of partial replacement of brown fish meal (BFM) by fermented soybean meal (FSBM) in diets of Chinese soft-shelled turtle (Pelodiscus sinensis). The turtles (initial mean body weight, (115.52 ± 1.05) g) were fed with three experimental diets, in which 0%, 4.72% and 9.44% BFM protein was replaced by 0%, 3% and 6% FSBM, respectively. Results showed that the feeding rate (FR), specific growth rate (SGR) and feed efficiency ratio (FER) of turtles fed with the diet containing 3% FSBM were not significantly different from the control group (0% FSBM) (P > 0.05). However, FR, SGR and FER of turtles fed with the diet containing 6% FSBM were significantly lower than those of the control group (P < 0.05). No significant differences were observed in the activities of serum glutamic-oxalacetic transaminase and glutamicpyruvic transaminase among dietary treatments (P > 0.05). However, the uric acid concentration in turtles fed with the diet containing 3% or 6% FSBM was significantly lower than that in the control group (P < 0.05). There were no significant differences in the activities of lysozyme, alkaline phosphatase and total superoxide dismutase among dietary treatments (P > 0.05). The results suggested that FSBM could replace 4.72% BFM protein in turtle diets without exerting adverse effects on turtle growth, feed utilization and measured immune parameters.

Physiological responses in vitamin C system during hibernation in juvenile Chinese soft-shelled turtle Pelodiscus sinensis

Vitamin C (Vc) is an important antioxidant that helps turtles tolerating stressful environment. This work quantified changes in tissue Vc levels during winter hibernation in the soft-shelled turtle Pelodiscus sinensis , to reveal the stress response pattern of tissue Vc during hibernation and contribute basic data for turtle culture. We sampled juvenile soft-shelled turtles at pre-hibernation (17.0°C mud temperature;MT), during hibernation (5.8°C MT) and after arousal (20.1°C MT) in the field. The transcript levels of the gene encoding L-gulonolactone oxidase (GLO), the key enzyme for Vc synthesis, decreased significantly during hibernation. However, GLO activity did not match the GLO transcription patterns and remained stable during hibernation, and showed temperature-dependent kinetic characteristics. Vitamin C levels in the brain, liver, kidney, and spleen (but not muscle) all decreased significantly during hibernation, but recovered to pre-hibernation levels or even higher levels after arousal. The soft-shelled turtle endured 5 months of hibernation with no significant oxidative damage in most tissues, except in the spleen. Splenic Vc was nearly exhausted during hibernation, accompanied by a significantly elevated malonaldehyde (MDA) level. Although the high level of oxidative damage quickly decreased after arousal, the potential tissue damage in the spleen during hibernation might account for the weakened immune capacity of turtles after hibernation.

Effects of ration level and feeding frequency on digestibility in juvenile soft-shelled turtle, Pelodiscus sinensis

The effects of ration level and feeding frequency on digestibility in juvenile soft-shelled turtle, Pelodiscus sinensis, were investigated. Four ration levels 1.5%, 2.5%, 4.0% and satiation （6.0% BW/d） were used. Apparent digestibility （AD） of dry matter （DMAD）, protein （PAD） and protein real digestibility （PRD） were significantly affected by ration level, but not by feeding frequency when the ration level was similar. However, the feeding frequency affected the AD, DMAD, PAD and PRD significantly when the turtles were fed to satiation. The relationship between fecal protein content （Y） and protein intake （At） can be expressed as a quadric equation： Y=-0.1742＋0.1476X-0.0003X^2 （r^2=0.876, n=27, F=93.92, P〈0.01）.

Effects of Dietary Soy Protein Concentrate on Growth, Digestive Enzymes Activities and Target of Rapamycin Signaling Pathway Regulation in Juvenile Soft-Shelled Turtle, <i>Pelodiscus sinensis</i>

Soft-shelled turtle, Pelodiscus sinensis is important aquatic species in China, and searching for alternatives protein resources to fish meal (FM)-based feeds in feed has become urgent and important for its sustainability development. The present study was conducted to assess the effects of dietary soy protein concentrate (SPC) on growth, digestive enzymes and target of rapamycin (TOR) signaling pathway of juvenile P. sinensis (4.56 ± 0.09 g). SPC was applied to replace FM protein at 0%, 15%, 30% and 60% (designated as T0, T15, T30 and T60, respectively), and each diet was fed to triplicate groups. The results showed that there was no significant difference in growth performance and feed utilization except of the turtles fed with T60 diet, of which showed poorer daily weight gain and feed conversion rate. The pepsin/trypsin and Na+-K+ ATP-ase activities decreased dramatically when SPC level increased, and lipase activities in liver and intestinal tract also showed decline tendency. However, amylase activities were unaffected. No significant differences were observed in TOR, S6K1 and 4E-BP1 genes mRNA expression level of TOR signaling pathway among the treatments. However, the relative phosphorylated level of these proteins decreased significantly when SPC level increased. The present study indicated that high SPC substitution level would suppress digestive enzymes and TOR signaling pathway proteins phosphorylated level and eventually result in growth reduction of P. sinensis.

Discovery of virus pathogen from soft-shelled turtle Trionyx sinesis

THE soft-shelled turtle Trionyx sinesis, because of its nutritional and medicinal values,

Effects of acute cold exposure on oxidative balance and total antioxidant capacity in juvenile Chinese soft-shelled turtle,Pelodiscus sinensis

Acute cold exposure may disturb the physiological homeostasis of the body in ectotherms.To date,there has been no information on the effects of cold exposure on homeostasis of reactive oxygen species(ROS)or antioxidant defense response in the Chinese soft-shelled turtle,Pelodiscus sinensis.In this study,P.sinensis juveniles were acclimated at 28℃,transferred to 8℃ as cold exposure for 12 h,then moved back to 28℃ rewarming for 24 h.We measured the ROS level and total antioxidant capacity(TAC)in the brain,liver,kidney and spleen at 2 and 12 h cold exposure,and at the end of the rewarming period.Malonaldehyde(MDA)and carbonyl protein were used as markers of oxidative damage.Turtles being maintained simultaneously at 28℃ were used as the control group.Cold exposure did not disturb the ROS balance in all 4 tissues,while rewarming raised the ROS level in the brain and kidney of P.sinensis.Cold exposure and rewarming decreased the TAC in the brain,liver and spleen but did not change the TAC in the kidney.MDA and carbonyl protein levels did not increase during the treatment,indicating no oxidative damage in all 4 tissues of P.sinensis.Our results indicated that extreme cold exposure did not impact the inner oxidative balance of P.sinensis,but more ROS was produced during rewarming.P.sinensis showed good tolerance to the harsh temperature change through effective protection of its antioxidant defense system to oxidative damage.This study provides basic data on the stress biology of P.sinensis.

Supplementation with turmeric residue increased survival of the Chinese soft-shelled turtle (Pelodiscus sinensis) under high ambient temperatures

Turmeric residue（TR）, containing residual levels of curcumin, is a solid by-product waste generated after the extraction and separation of curcumin from turmeric root. A feeding trial was conducted to evaluate the effects of TR on the survival of Chinese soft-shelled turtles（SSTs）, Pelodiscus sinensis, under a high ambient temperature. A total of 320 female SSTs were assigned randomly to two diets： basal diet（the control group, n=160） and an interventional diet supplemented with 10% TR（the TR group, n=160）. Our results demonstrated that supplementation of TR increased the SST survival rate by 135.5%, and superoxide dismutase（SOD） activity of SST liver by 112.8%, and decreased the malondialdehyde（MDA） content of SST liver by 36.4%, compared to the control group. The skin of the SST fed TR showed a golden color. High-performance liquid chromatography（HPLC） analysis indicated that the concentrations of curcumin in TR and the skin of the SST fed TR were（1.69±0.30） and（0.14±0.03） μg/g, respectively. Our observation suggests that supplementation of TR increased the survival rate of SST under high ambient temperatures. We speculated that the increased survival rate and tolerance at the high ambient temperature were associated with the anti-oxidation activity of curcumin from TR. Moreover, curcumin in TR could be deposited in SST skin, which made it more favored in the market of China. Our findings provide new knowledge and evidence to effectively reuse TR as a feed additive in animal and aquatic farming.

Division of Chinese soft-shelled turtle intestine with molecular markers is slightly different from the morphological and histological observation

The Chinese soft-shelled turtle(Pelodiscus sinensis)is a commercially important species in Asian countries.Knowledge of its nutritional requirements and physiology is essential for determining the appropriate content of the feed for this animal.However,the lack of functional characterization of the intestine of this turtle limits the understanding of its absorption and utilization of nutritional materials.To solve this problem,this work utilized anatomical and histological methods to characterize 9 segments sampled along the anterior-posterior axis of the intestine.Furthermore,9 genes,which have been well documented in the intestine division of mammals and fish,were employed to functionally characterize the 9 sampled segments.Our results suggest that regions covering from the starting site to S3(position at 29.9%of the total length from the starting of the intestine)are the equivalent of mammalian dedumonen,and those covering S4(40.2%)and S5(65.4%),posterior to S8(92.7%),are the equivalent of the mammalian ileum and the large intestine,respectively.As to the region spaning S6(81.3%)and S7(87.3%),its functional equivalent(small intestine or large intestine)may be variable and depends on the functional genes.This molecular characterization in relation to the division of the intestine of Chinese soft-shelled turtle may contribute to the understanding of the nutritional physiology of the turtle,and promote Chinese soft-shelled turtle production.

Effect of localized oil spills on Atlantic loggerhead population dynamics

The purpose of this study is to analyze the population dynamics of loggerhead sea turtles (Caretta caretta) affected by localized oil spills. Methods include development of a spatial, stage-classified matrix model parameterized for the following primary loggerhead populations: Gulf of Mexico, western North Atlantic Ocean, and Florida peninsula. Oil spills are simulated deterministically in each population's nesting region, with 1) oil-induced mortality ranging from 25% to 100% and 2) stage classes affected either proportionally or equally. A transient sensitivity analysis was performed to determine the parameters most influential to the population growth rate. Results suggest that increased protection and understanding of young sea turtles found in the Sargasso Sea is essential to the survival of the species. In addition, findings provide insights into the population dynamics of the At- lantic loggerhead turtles and identify conservation measures appropriate in each oil spill case.

Medicinal Soft-Shelled Turtle

Ingredients:Whole soft-shelled turtle; Chinese wolf-berry fruit(Lycium Chinese), milk vetch membrane root(Astragalus membranaceus), Chinese angelica andAmerican ginseng--about 25g of

超临界CO_2萃取甲鱼油的研究

用超临界CO2 萃取技术从甲鱼中提取分离甲鱼油 ,着重探讨了影响萃取率的主要因素 ,并同传统的溶剂萃取法进行比较 ,分析了两种萃取方法得到甲鱼油的脂肪酸组成。结果表明 :超临界CO2 萃取甲鱼油的适宜工艺条件为 :2 0MPa ,4 5℃ ,CO2 流量为 1.8kg/ (h·g)原料 ,萃取时间为 6h ;超临界CO2 萃取法萃取率 (98.1% )明显高于乙酸乙酯萃取法 (85 .2 % ) ,两种方法所得到的甲鱼油主要脂肪酸组成基本一致 ,甲鱼油中含有的不饱和脂肪酸高达 70 %以上 ,其中多不饱和脂肪酸含量大于 2 6 %。

甲鱼油中多不饱和脂肪酸的提取与纯化研究

本文研究了甲鱼油中多不饱和脂肪酸的提取与纯化。采用乙酯化法提取甲鱼油中的多不饱和脂肪酸,通过正交实验确定了最佳提取条件:乙醇/甲鱼油比0.73:1,NaOH用量3%(油重),反应温度65℃,反应时间2h;采用尿素包合法纯化多不饱和脂肪酸,考察尿素用量、搅拌时间、包合次数对尿素包合率的影响,通过正交实验确定最佳纯化条件:尿素/乙醇饱和溶液:脂肪酸乙酯=15:1,搅拌时间3h,包合次数1次。实验结果表明,应用乙酯化法与尿素包合法可从甲鱼油中制得高纯度的多不饱和脂肪酸。

甲鱼油脂肪酸组成的研究

用热提取方法抽取出甲鱼油，用ＫＯＨ－ＣＨ３ＯＨ酯交换法甲酯化处理，以色谱／质谱方法测定脂肪酸的组成。共鉴定出２１种脂肪酸，其中以棕榈酸、棕榈油酸、硬脂酸、油酸、亚油酸、二十碳五烯酸（ＥＰＡ）。

甲鱼油中ω-3多不饱和脂肪酸的提取与分析

目的为从甲鱼脂肪中提取甲鱼油,并分析其浓度及纯度。方法采用超临界CO2萃取技术,利用低温浓缩工艺富集甲鱼油中DHA和EPA。以超临界提取甲鱼脂肪为原料,用低温浓缩法富集DHA和EPA;从甲鱼脂肪中提取甲鱼油,然后用GC法测定DHA和EPA,并对不同温度、溶剂配比参数进行比较。结果利用超临界CO2萃取技术可以从甲鱼脂肪中获得CO2超临界萃取原油(SFE原油),并经正交分析得到最佳的组合为A1B3C3,其最优化工艺条件是:压力20MPa,温度50℃;时间4h,CO2用量为2m.lg-1,EPA和DHA的最高萃取率为:16.32%,SFE原油∶溶剂为1∶12、冷冻8h的条件下,得到的甲鱼油中EPA+DHA含量为26.39%。结论超临界CO2萃取技术简便易行,还具有安全可靠,无污染等优点。因此该方法为开发和综合利用甲鱼资源开辟了新的应用途径。

甲鱼蛋粉混合物对高脂模型大鼠血脂的影响

目的 研究甲鱼蛋粉混合物 (红花油、维生素E、大蒜粉、甲鱼蛋粉 )对高脂血症大鼠血脂的调节作用。方法 将 5 0只健康雄性SD大鼠按体重和血清胆固醇水平随机分成 5组 ,分别接受正常饲料、高脂饲料、高脂饲料加甲鱼蛋粉混合物 (低、中、高剂量分别为 0 5、1 5、5 0g kgBW )灌喂处理 ,7周结束 ,测定血清胆固醇 (TC)、甘油三酯 (TG)低密度脂蛋白胆固醇 (LDL C)和高密度脂蛋白胆固醇 (HDL C)。结果 与高脂饲料组相比 ,甲鱼蛋粉混合物能降低高脂血症大鼠TC、TG和LDL C ,其中高剂量组具有显著性 (P <0 0 5 )。结论 研究结果表明甲鱼蛋粉混合物具有降低高脂血症大鼠血脂的作用。

高效液相色谱法测定甲鱼油中EPA、DHA的含量

建立分离检测广西黄沙鳖甲鱼油中的EPA和DHA的高效液相色谱法。试验采用岛津VP-ODS色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇和水(体积比70∶30),紫外检测波长为254 nm,流速为0.7 mL/min,柱温为30℃。甲鱼油经0.5 mol/L KOH-乙醇溶液进行皂化,三乙胺-溴苯乙酮酯化后直接上机分析。结果表明:EPA、DHA在0.025 mg/mL^0.15 mg/mL范围内线性关系良好,R2EPA为0.999 1、R2DHA为0.999 5;该法用于甲鱼油的检测,回收率EPA为96.00%～98.67%,DHA为92.00%～98.00%,该法简便、快速、重现性好。

养血润肤饮联合海贽油治疗银屑病血燥证45例临床观察

目的:观察养血润肤饮联合海贽油治疗银屑病血燥证的临床治疗效果。方法:按照随机数字的方法将90例寻常型银屑病血燥证患者分为治疗组45例、对照组45例。治疗组予养血润肤饮口服联合海贽油外用,对照组予润燥止痒胶囊口服联合医用凡士林外用。观察2组PASI评分、临床治疗效果、中医症候评分情况并分析治疗前患者皮损面积、皮损严重程度与患者焦虑程度之间的线性关系。结果:2组治疗后PASI评分及临床治疗效果较本组治疗前降低(P<0.05),治疗组较对照组改善明显,组间比较差异具有统计学意义(P<0.05);中医症候评分较本组治疗前降低(P<0.05),治疗组较对照组改善明显,组间比较差异具有统计学意义(P<0.05),说明治疗组治疗效果优于对照组;患者皮损面积、皮损严重程度与患者焦虑程度均有相关性(P<0.05)。结论:养血润肤饮联合海贽油治疗银屑病血燥证具有较好的临床治疗效果。

甲鱼油中不饱和脂肪酸的测定

本文用气相色谱法测定了甲鱼油中各种脂肪酸的含量,运用色谱质谱联用分析对各种脂肪酸进行定性分析.发现重量不同的甲鱼油中所含C_(20)和C_(22)的多不饱和脂肪酸有较大的差异.

水产全价饲料及饲用油脂的品质评价

在湖南益阳随机抽取了4种市售甲鱼全价饲料和6种草鱼全价饲料,分析了其中的营养成分与挥发性盐基氮,抽取了3种饲用油脂,分析了其酸值和过氧化值。结果表明,4种甲鱼全价配合饲料中水分、粗灰分、粗蛋白、粗脂肪、粗纤维、总磷、钙的平均含量分别为5.49%、15.88%、40.64%、6.09%、1.90%、2.42%、4.28%,总挥发性盐基氮的含量为28.01 mg/100 g;6种草鱼全价配合饲料中水分、灰分、粗蛋白、粗脂肪、粗纤维、总磷、总钙的平均含量分别为9.34%、10.98%、29.41%、8.72%、12.01%、1.26%、0.85%;饲用油脂的平均酸值和过氧化值分别为11.70 mg/g和11.10 mmol/g。