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.

Measurement of Liver Size and Intrahepatic Blood Flow of Red-eared Turtles by Color Doppler Ultrasound

In the study, a total of 20 red-eared turtles （ half male and half female） were selected to conduct the measurement of liver size and intrahepatic blood flow of red-eared turtles by color Doppler ultrasound. The results showed that the fight hepatic lobe could be scanned through the right carotid anterior acoustic win- dow, and the left hepatic lobe could be scanned through the left carotid anterior acoustic window, but the vision would be obstructed by the air in trachea. The liver could also be scanned through the left femoral anterior acoustic window and the fight femoral anterior acoustic window when filling of bladder was good. The correla- tion regression analysis suggested that estimated values of liver showed no linear relationship with weight, the longest back curve and the widest back curve. Through the study, the normal indicators for ultrasound examination of red-eared turtle liver were established, in order to provide a reference for examination of turtle liver.

基于“通权达变”思想探究柴胡不同炮制方法

目的基于“通权达变”思想分析柴胡不同炮制品的古今用药经验及现代研究进展,为柴胡炮制机制及炮制工艺研究提供参考。方法从中华医典、读秀等数据库系统挖掘柴胡炮制方法的历史演变及柴胡饮片临床用药特色;通过查阅文献梳理柴胡不同炮制品的现代研究进展。结果历代医药典籍记载柴胡炮制品种丰富,不同炮制品功效上存在较明显差异:醋柴胡疏肝解郁之效增强,升散之性减弱;鳖血柴胡滋阴退热之效增强,浮阳之性减弱;酒柴胡舒筋通络、活血止痛之效增强。现代研究对柴胡不同炮制品的科学内涵进行了不同程度解读,但仍存在一些问题,如炮制工艺参差不齐、质量标准尚未统一等。结论柴胡药用历史悠久,临床应用广泛,炮制品种较多,亟需加强不同炮制品的药效物质基础及炮制机理的系统研究,加深对柴胡不同炮制品科学内涵认识,建议规范炮制工艺和制定更加科学合理的质量标准,为临床提供质量稳定可控、品种丰富的柴胡炮制品,发挥柴胡临床治疗的特色和优势。

改良式鳖甲灸循经推熨治疗寒凝血瘀证原发性痛经患者的临床效果观察

目的探讨改良式鳖甲灸循经推熨治疗寒凝血瘀证原发性痛经患者的临床效果。方法选取2020年8月至2021年8月阜阳市妇女儿童医院收治的60例寒凝血瘀证原发性痛经患者作为研究对象,依照随机数字表法将其分为试验组和对照组,每组30例。对照组给予布洛芬颗粒治疗,试验组在对照组的基础上另给予改良式鳖甲灸循经推熨治疗,2组患者均治疗3个月经周期。比较2组患者中医症状评分、疼痛持续时间、疼痛视觉模拟量表(VAS)评分、临床疗效、血清前列腺素E2(PGE2)、前列腺素F2a(PGF2a)及不良反应。结果治疗后2组患者小腹冷痛、畏寒肢冷、腰骶酸痛、经色黯而有瘀块评分较治疗前均降低,且试验组上述评分均低于对照组,差异均有统计学意义(P<0.05);治疗后2组患者疼痛持续时间较治疗前均缩短,且试验组短于对照组,差异均有统计学意义(P<0.05);治疗后2组患者VAS评分较治疗前均降低,且试验组低于对照组,差异均有统计学意义(P<0.05);试验组总有效率高于对照组,差异有统计学意义(P<0.05);治疗后2组患者血清PGE2水平较治疗前均升高,且试验组高于对照组,差异均有统计学意义(P<0.05);治疗后2组患者血清PGF2a水平较治疗前均降低,且试验组低于对照组,差异均有统计学意义(P<0.05);2组患者均未出现胃肠道反应、出血等不良反应。结论改良式鳖甲灸循经推熨治疗寒凝血瘀证原发性痛经患者能够改善临床症状,疗效确切,同时能够调节前列腺素水平,安全可靠。

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

Box-Behnken响应面实验优化鳖血冻干粉的制备工艺

目的优化鳖血冻干粉的制备工艺,为提高鳖血的服用便捷性和质量稳定性提供参考。方法采用真空冷冻干燥技术制备鳖血冻干粉。以溶解度为考察指标,通过单因素实验和Box-Behnken响应面实验优化鳖血冻干粉的冷冻干燥工艺参数,并验证。结果鳖血冻干粉最优冷冻干燥工艺为预冻时间4 h、干燥时间13 h(0℃前)、解析干燥温度25℃。3批按最优工艺制备的鳖血冻干粉的平均溶解度为95.72%(RSD=0.68%,n=3),与理论溶解度(96.66%)的相对误差为-0.97%。结论本研究优化的冷冻干燥工艺稳定可行,所制样品的溶解度较高。

THE USE OF SNAPPING TURTLE RED BLOOD CELLS FOR STANDA RDIZATION IN FLOW CYTOMETRY

Science flow cytometry measurement of intensity is relative measur, ement. The standards are therefore necessary. There are two types of standards. One is the instrumental standard for adjustment FCM. Fluorescent microspheres or glutaraldehyde fixed chicken red blood cells are commonly used for this purpose. The other is the DNA internal standard of nuclear DNA ploidy analysis for correcting the changes in preparation and staining condition and instrument drift during measurement.

鳖血提取物对小鼠免疫功能正向调节作用的研究

目的 :研究鳖血提取物对小鼠的免疫调节功能。方法 :采用环磷酰胺为免疫抑制剂 ,获得免疫功能低下的小鼠动物模型。比较鳖血提取物作用前后 ,小鼠T细胞亚群的数量、NK细胞的杀伤功能 ,淋巴细胞的增殖功能以及细胞因子水平的变化。结果 :鳖血提取物对免疫功能低下小鼠的CD4 + T细胞在外周血中的比例、NK细胞的杀伤活性、淋巴细胞的增殖功能 ,均具有正向调节作用 (P <0 .0 5 ) ,并呈剂量依赖性 ;能提高淋巴细胞分泌IFN γ的能力。结论

中华鳖外周血细胞形态学观察

用光镜和电镜,观察了中华鳖外周血细胞的形态学与组织化学的特性。 以瑞氏-姬姆萨混合液染色的血涂片,在光镜下可见到红细胞、单核细胞、嗜异性细胞、淋巴细胞、嗜酸性细胞、嗜碱性细胞、血栓细胞和浆细胞;此外。还可以看到极少数成红细胞、异形红细胞、嗜碱性点彩和无定形核物质。除红细胞、嗜酸性细胞与血栓细胞外,各种细胞的超微结构基本上与人相应细胞的大同小异。

龟鹿胶、淫羊藿及红景天对骨质疏松大鼠骨密度及破骨细胞的影响

背景:中药研究骨质疏松症目前缺乏系统的药效学研究,中药组方尚欠精简,机制研究与中医理论指导不够紧密。目的:观察中药复方龟鹿胶、淫羊藿、红景天对去卵巢大鼠血清各指标、骨密度及破骨细胞的的调控作用。方法:建立去卵巢大鼠模型,分别用自拟中药组方龟鹿胶组、淫羊藿组、红景天组进行灌胃,以假手术组和模型组进行对比。造模后第4周开始称体质量,每周1次。造模后12周后测定大鼠骨密度、血清中钙、磷含量、血碱性磷酸酶活性等指标比较治疗效果,应用体外培养破骨细胞观察各组中药对破骨细胞抑制率的影响。结果与结论:龟鹿胶能显著减少去卵巢大鼠体质量的增加幅度(P<0.05)。相比模型组3种中药治疗组大鼠血清中钙含量明显升高(P<0.05),血磷与碱性磷酸酶活性明显降低(P<0.01或P<0.05)。龟鹿胶组和淫羊藿组大鼠骨密度显著高于模型组(P<0.05)。3种中药治疗组与假手术组相比吸收陷窝数明显减少(P<0.05),其中红景天组吸收陷窝数减少较显著(P<0.01),红景天组对破骨细胞吸收功能具有抑制作用,抑制率为86.85%(P<0.01)。结果证实,中药组方龟鹿胶、淫羊藿、红景天对去卵巢骨质疏松均具有显著疗效,其中龟鹿胶组和淫羊藿组主要以增加骨密度为主,红景天组具体表现在对破骨细胞吸收功能具有抑制作用。

凉血解毒散对鳖红脖子病的临床治疗

鳖的红脖子病是由嗜水气单胞菌引起的一种急性传染病 .本试验运用中医学理论对其进行辨证施治 ,组方凉血解毒散应用于预防和临床治疗 ,获得了很好的治疗效果 .证明该方与抗菌素一样能有效地防治鳖的红脖子病 .并通过抑菌试验 。

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。