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.

Effects of Pesticide Exposure on Embryonic Development and Hatchling Traits of Turtles

Deltamethrin is a widespread environmental hormone with endocrine-disrupting properties, but its effect on embryonic development of reptiles is largely unexplored. We investigated the effects of deltamethrin on embryonic development and offspring traits in two turtle species, one with parchment-shelled eggs and the other with rigidshelled eggs. Deltamethrin exposure during egg incubation did not affect hatching success and hatchling body size in either species. However, embryonic exposure to deltamethrin resulted in reduced hatchling locomotor performance in the red-eared slider turtle（Trachemys scripta） with parchment-shelled eggs, but not in the Chinese three-keeled pond turtle（Chinemys reevesii） with rigid-shelled eggs. These results suggest that parchment-shelled eggs are likely more vulnerable to deltamethrin than rigid-shelled eggs.

Effect of Incubation Temperature on the Somatic Development of the Snail-Eating Turtle Malayemys macrocephala

The rising average global temperature can lead to changes both in the physical and biological environments and affect the survival of organisms. Freshwater turtles are considered to be susceptible species since their development is dependent on incubation temperature. In Thailand, although several species of freshwater turtle are present, the extent of their susceptibility to temperature change is unknown due to the lack of information on their developmental patterns. This research, therefore, is aimed at examining the effects of temperature on somatic development in Malayemys macrocephala, a native species and the most common freshwater turtle in Thailand. Turtle eggs were collected from rice fields in the central part of Thailand during December 2011 to February 2012 inclusive. Eggs （237-238 per condition） were incubated in microprocessor-controlled incubators at three different temperatures （26 ℃, 29℃ and 32℃） with a relative humidity in excess of 80%. Each week, until the remaining eggs hatched, eggs were randomly selected, removed and dissected to reveal the developing embryo to screen for developmental stage and any abnormalities. The incubation period （lay to hatch） was not significantly different among the three temperatures （115 ±11.3 d, 115 ±20.3 d and 109±17.8 d, respectively）, but the growth patterns, as indicated by the stages of development and carapace lengths, were significantly different. At a high temperature （32℃）, turtle embryos showed a significantly faster growth rate than at the intermediate and low temperatures （29 ℃and 26 ℃）, but had a significantly （over 3.3-fold） higher incidence of developmental abnormalities （especially deformed bodies） than at the lower temperatures. Overall, the results indicate that incubation temperature is an important variable affecting the somatic development of this tropical freshwater turtle species, whilst abnormalities in the embryonic body may be a sensitive indicator of extreme thermal stress.

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.

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

中华鳖卵孵化过程中物质和能量的动态

在温、湿度分别为 30℃、 - 2 2 0kPa条件下孵化中华鳖 (Pelodiscussinensis)卵 ,以检测孵化过程中胚胎生长及对物质和能量的利用。孵化过程中 ,每隔 7天测定卵重。孵化第 10天始 ,每隔 5天解剖 15枚卵 ,分离为胚胎、卵黄和卵壳。初生幼体称重后冰冻处死 ,随后解剖分离为躯干、剩余卵黄和脂肪体。所有样品 6 5℃恒温干燥 ,用索氏抽提仪测定脂肪含量 ,氧弹式热量计测定能量含量 ,马福炉测定无机物含量。本研究中华鳖卵的平均孵化期为 45 5天。卵重量在孵化过程中基本维持恒定 ,表明中华鳖卵与环境的水分交换不明显。孵化第0～ 2 0天、 2 0～ 30天、 30～ 40天和 40～ 45天 ,胚胎分别利用新生卵内总能量的 6 %、 2 4%、 5 9%和 4%。 0～ 30天 ,胚胎生长缓慢 ;30～ 40天 ,胚胎生长最迅速 ;孵化末期胚胎生长减缓。胚胎发育所需的无机物来自卵黄和卵壳 ;卵壳提供的无机物约占初生幼体无机物总量的 13 5 %。孵化过程中 ,干物质、脂肪和能量的转化率分别为 79 6 %、 5 8 7%和 6 6 5 %。初生幼体的能量组成为 :躯干占 71 8% ,脂肪体占 19 4% ,剩余卵黄占 8 8%。

中华鳖蛋营养价值评价

本试验旨在对中华鳖蛋的营养价值进行评价。选取日本种群和黄河种群中华鳖蛋各200枚,分别进行基本营养成分、胆固醇、氨基酸与脂肪酸含量检测,以确定中华鳖蛋的营养价值。结果表明:1)2个种群中华鳖蛋蛋清中水分、粗灰分、粗蛋白质和粗脂肪含量分别为94.92%和95.18%、0.59%和0.37%、3.26%和3.20%、0.86%和0.82%;蛋黄中水分、粗灰分、粗蛋白质和粗脂肪含量分别为67.81%和69.01%、1.85%和1.98%、21.22%和21.15%、10.47%和8.91%。2)水解氨基酸含量检测结果显示,2个种群中华鳖蛋蛋清与蛋黄中均检出17种水解氨基酸,且均以谷氨酸含量最高,天冬氨酸含量次之;蛋清中氨基酸总量分别为557.61、461.43 mg/g,必需氨基酸总量分别为212.90、175.01 mg/g,呈味氨基酸总量分别为213.49、176.61 mg/g;蛋黄中氨基酸总量分别为567.72、536.04 mg/g,必需氨基酸总量分别为215.73、203.09 mg/g,呈味氨基酸总量分别为212.43、201.50 mg/g。必需氨基酸组成评价分析显示,2个种群中华鳖蛋蛋清和蛋黄中除蛋氨酸+半胱氨酸百分比外,其余必需氨基酸百分比均超过人体蛋白质氨基酸模式谱的参考值,属于优质蛋白质。游离氨基酸含量检测结果显示,2个种群中华鳖蛋蛋清中氨基酸总量分别为23.38、26.08 mg/kg,呈味氨基酸总量分别为12.12、12.63 mg/kg,分别占氨基酸总量的52%和48%;蛋黄中氨基酸总量分别为11.13、13.53 mg/kg,呈味氨基酸总量分别为3.67和4.02 mg/kg,分别占氨基酸总量的33%和30%。3)2个种群中华鳖蛋蛋黄中饱和脂肪酸总量分别为40.83%和44.85%,不饱和脂肪酸总量分别为56.80%和50.51%,胆固醇含量分别为10.49和10.84 mg/g。由此可见,中华鳖蛋水分、粗蛋白质含量高,粗脂肪含量低,必需氨基酸组成满足理想氨基酸模式,呈味氨基酸含量高,胆固醇含量低,是一种口感鲜美及营养价值较高的蛋类。

温度对中华鳖卵孵化和胚胎发育的影响

在 2 5℃、 2 7 5℃、 30℃、 32 5℃和 35℃条件下 ,分别研究了中华鳖 (Trionyxsinensis)卵的孵化和胚胎的发育。卵黄和卵壳的湿重分别占卵重的 85 10 %和 14 90 %,含水量分别为 76 6 3%和 2 1 0 1%。中华鳖胚胎重随孵化时间呈逻辑斯谛曲线增长 ,孵化温度越高 ,胚胎发育速度r越大。 2 5℃～ 30℃的胚胎发育仅表现为‘S 型生长曲线的前、中段 ,在孵化后期发育速度未受到明显的抑制 ,32 5℃和 35℃温度条件下 ,后期发育速度受到了较大的影响。孵出稚鳖湿重和干重均与初始卵重存在极显著的线性相关关系。孵化温度对孵出稚鳖湿重没有显著影响 ,但对孵出稚鳖干重的影响极显著。随孵化温度的升高 ,孵出稚鳖干重呈‘U 形变化 ,在适中温度下 ,胚胎的生长效率最低。卵黄干重随孵化时间呈指数递减 ,温度越高 ,卵黄的利用速度越快。在孵化过程中 ,卵重、卵干重和卵内水总量都随孵化时间的延长、孵化温度的上升而略有下降。孵化期随温度升高呈双曲线缩短 ,其相关关系可用方程I =30 6 +10 9/ (T - 2 2 5 )来表示。中华鳖卵的最适孵化温度在 32 5℃左右 ,胚胎在孵化后期比早期对温度的需求可能要低一些。

甲鱼蛋粉调节大鼠血清胆固醇的基础研究

目的 研究甲鱼蛋粉对高胆固醇血症大鼠血清胆固醇的调节作用。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 ,每组 10只 ,分别接受正常饲料、高脂饲料、高脂饲料加甲鱼蛋粉灌胃 (低、中、高剂量分别为 0 75、1 5 0、3 0 0 kgBW)处理 ,实验周期为 2 4周 ,实验期末 ,收集大鼠粪便 ,股动脉取血 ,酶法测定血清胆固醇 (TC)、高密度脂蛋白胆固醇 (HDL C)和低密度脂蛋白胆固醇 (LDL C)的含量 ,气相色谱法检测粪便胆固醇、粪甾醇、粪甾烯 ,循环酶法测定总胆汁酸的含量。结果 与高脂饲料组相比 ,中剂量和高剂量的甲鱼蛋粉均能显著降低 (P <0 0 5 )高胆固醇血症大鼠血清TC ,且剂量为 1 5 0g kgBW时能显著降低血清LDL水平 ;两剂量组均显著增加粪便胆固醇、粪甾醇和粪甾烯以及胆汁酸的排出。

孵化温度对乌龟胚胎能量利用及矿物质代谢的影响

在24℃、27℃、30℃和33℃条件下孵化乌龟(Chinemysreevesii)卵,检测温度对胚胎利用卵内能量和矿物质的影响。孵化温度显著影响乌龟胚胎的能量利用。卵在温和温度下(27℃和30℃)比在高温(33℃)和低温(24℃)孵化有较高的干物质、脂肪和能量转化率。因而,27℃和30℃新生幼体的总能量高于33℃和24℃下孵出的幼体。新生幼体内剩余卵黄随温度升高而增大。27℃下新生幼体的躯干能量显著高于其它温度。此外,孵化温度显著影响乌龟胚胎钙代谢,但对镁和钾代谢影响微弱。24℃下胚胎从卵黄和卵壳吸收的总钙量小于其它温度条件下胚胎,与之相对应,24℃下孵出卵卵壳内残留较多的钙。由此可见,温和温度(27-30℃)可提高胚胎能量转化效率,促进胚胎从卵黄和卵壳吸收较多的矿物质。

孵化温度对中华鳖胚胎物质和能量利用的影响

用多重温度组合 [7个恒温 ( 2 3、 2 4、 2 7、 2 8、 3 0、 3 3和 3 4℃ )和 1个波动温度 ( 2 2 3～ 3 2 8℃ ) ]孵化中华鳖 (Pelodiscussinensis)卵 ,检测温度对胚胎利用卵内物质和能量的影响。卵在波动温度和温和温度下( 2 7℃和 2 8℃ )孵化有较高的干物质、脂肪和能量转化率 ,孵出幼体因而含有较多的无机物 ,幼体躯干和脂肪体含有较高的能量。卵在极端高温 ( 3 4℃和 3 3℃ )或极端低温 ( 2 3℃ )下孵化物质和能量转化率较低 ,使得孵出幼体中无机物较少 ,幼体躯干和脂肪体中含能量较低。孵化温度显著影响中华鳖孵出幼体内的能量分配 :2 7℃和 2 8℃孵出幼体含能量较高且相似 ,但 2 7℃孵出幼体剩余卵黄能量小于 2 8℃孵出幼体 ;3 4℃和 2 3℃孵出幼体含能量较低且相似 ,但 2 3℃孵出幼体剩余卵黄能量大于 3 4℃孵出幼体。

环境因子对龟鳖动物胚胎发育影响的研究进展

对温度、湿度、孵化介质三种环境因子对龟鳖动物胚胎发育影响的研究进展进行了综述,旨在为今后研究环境因子对龟鳖动物胚胎发育的影响机制和提高龟鳖动物的人工孵化率提供一些理论依据。

不同孵化湿度下的乌龟卵孵化成功率及新生幼体特征

A total of 51 Chinese three-keeled pond turtle Chinemys reevesii egg s were incubated on wet and dry substrates (water potentials of -12 kPa and -3 00 kPa, respectively) at 30 ℃ to assess the influence of the hydric environment s on egg survival and hatchling traits. Eggs incubated in the dry condition lost weight, whereas those incubated in the wet condition did not. The hydric enviro nments did not affect incubation lengths (60 3 vs 60 7 days) as well as ha tching successes (69 2% vs 88 0%). Except for wet body mass and carapace width, which were larger for hatchlings from the -12 kPa treatment than from the -300 kPa tr eatment, the hydric environments did not affect most hatchling traits. These una ffected traits included dry body mass, carapace length, tail length, limb length , hatchling components (carcass, residual yolk and fat bodies), swimming perform ance and critical thermal minimum. In conclusion, within the water potential of -12 to -300 kPa, hydric environments have little effect on embryonic developme nt and hatchling traits in Chinese three-keeled pond turtles .