中华鳖外周血细胞病理显微结构研究

对病鳖血液细胞的显微观察发现,病鳖红细胞形态与正常鳖相比有较大差异,甚至有破碎现象;嗜中性粒细胞、单核细胞及血栓细胞在细胞形态上也有差异。     

中华鳖血细胞显微和超微结构的观察

关于中华鳖血细胞种类和结构目前尚无一致的结论。王石泉［１］认为鳖具有嗜天青粒细胞。蒋立科［２］认为嗜中性粒细胞即嗜天青粒细胞,而且鳖缺乏血栓细胞,可能存在其它凝血机制。鉴于此我们对鳖血细胞的显微形态和超微结构进行了观察,以资参考和比较。收稿日期：１９９８－０８－２４１　材料和方法鳖７只、体重６４～１１２ｇ,雌雄均有。左体动脉弓取血［３］,制作血涂片,Ｗｒｉｇｈｔ染色,显微镜油镜下观察、拍照、测量,至少２００个白细胞分类计数电镜标本制作,采用肝素抗凝血离心,取富含白细胞层,戊二醛锇酸双固定,梯度乙…     

EM技术在中华鳖控温养殖中的应用

EM是有效微生物群的英文缩写,它是日本琉球大学比嘉照夫教授用几十年时间研制出来的新型复合微生菌剂。这种菌剂是由酸杆菌,光合细菌、放线菌、酵母菌、乳酸菌5科10属80多株菌种复合培养而成,它不含任何化学物质,无毒副作用,不污染环境。日前应用较多的是在畜禽养殖上,其     

HACCP在中华鳖健康养殖中的应用

<正>HACCP是目前国际普遍认可的用于控制食品安全的先进质量管理体系,其原理在中华鳖健康养殖中的应用就是对养殖过程的每个环节进行危害分析,并确定关键控制点加以控制,使养殖生产免受可能发生的生物、化学、物理因素的危害,确保中华鳖质量安全。蚌埠     

新鲜虾壳粉在中华鳖饲喂中的应用

本研究将沅江市南大冷冻加工有限公司废弃的新鲜小龙虾虾壳粉碎成80目左右,部分替代目前市场中华鳖料中的高价值的进口鱼粉来生产“速冻鲜饲料”。且于2012年5月至10月在益阳沅江等地进行利用“速冻鲜饲料”进行的中华鳖养殖试验。结果表明,较之其他厂家生产的常规粉状饲料进行的中华鳖饲养,该饲料的研发减低中华鳖饲喂成本,同时利用了虾壳中虾青素的自然添加,有效增强了中华鳖的免疫机能,提高了中华鳖抗病性。通过该饲料一定时间段的饲喂,中华鳖从根本上改变品质,具有野生鳖形态、风味及营养价值。     

EM制剂在中华鳖温室养殖中的应用研究

本研究应用ＥＭ制剂进行全池泼洒,并添加于鳖饲料中进行近温养殖试验,参试池７－９号,其中９号为对照池,７、８号试验池,全池泼酒ＥＭ原露１０ｍｍｐ,饲料中添加１％ＥＭ原露,通过２５８天的饲养试验,结果表明７号、８号池日增重率分别比对照池提高１９．６％、１５．７％。结果还表明,ＥＭ制剂全池泼洒可改善水质,延长池水使用时间;ＥＭ制剂添加于鳖饲料能增强鳖体抗病能力,提高饲料利用率。     

两种酶制剂在中华鳖养殖中的应用研究

本研究应用两种不同酶制剂添加于鳖饲料中进行养殖试验，试验共分A．B、C、D和E组，其中A组为对照组，B组、C组和D组为酶制剂Ⅰ，添加量分别是0．1％、0．2％和0．3％。E组为酶制剂Ⅱ，添加量是0．2％。通过对71天的饲养试验，结果表明，B组、C组和E组的日增重率分别比对照组提高40．9％、29．6％和27．0％。而D组的日增重率反而比对照降低21．7％。结果还表明，酶制剂添加于鳖饲料能增强鳖的抗病能力，但会导致饲料效率降低。     

紫红笛鲷血细胞发生研究

将体质量510~620 g的紫红笛鲷用间氨基苯甲酸乙酯甲磺酸盐麻醉后解剖,取出头肾、体肾、肝脏、脾脏,用干净的刀片居中横切、纵切,印片和涂片用Wright-Giemsa染液染色10 min,观察头肾、体肾、脾脏、肝脏4个组织印片以及外周血涂片。试验结果显示,头肾、体肾、脾脏是其造血器官。头肾能发育生成各类型血细胞,体肾能生成红细胞、淋巴细胞、粒细胞和单核细胞,脾脏能生成淋巴细胞。肝脏中未发现幼稚型血细胞。红细胞发育经过原红细胞、早幼红细胞、中幼红细胞、晚幼红细胞和红细胞5个阶段;原粒细胞发育至早幼粒细胞后,分化为嗜碱性中幼粒细胞、嗜酸性中幼粒细胞和中性中幼粒细胞,最后发育成嗜碱性粒细胞、嗜酸性粒细胞和中性粒细胞;淋巴细胞和单核细胞发育经过原始、幼稚和成熟3个阶段。血细胞在发育过程中,胞体逐渐变小,细胞核逐渐变小,染色质由疏松到致密;粒细胞中,颗粒由少到多。     

三氯异氰尿酸在中华鳖病防治上应用研究

对三氯异氰尿酸进行了室内抑杀菌试验,其对中华鳖气单胞菌的MIC为0.25 mg/L,MBC为0.25～0.50 mg/L;急性毒性试验表明,该药对鳖的安全浓度为4.45 mg/L;对池水中细菌杀灭效果试验表明,药物有效浓度为0.5～1.0 mg/L;对池水亚硝酸盐氮（NO2-N）影响,1 mg/L TCCA分别作用24h、48 h,其对NO2^-N的消除率分别为9.50%和19.05%,同时进行了生产性防治试验,取得较好效果,为正确合理地使用该类药物提供参考依据.     

Growth, haematological parameters and tissue lipid peroxidation of soft-shelled turtles, Pelodiscus sinensis, fed diets supplemented with different levels of ferrous sulphate

Soft-shelled turtles, Pelodiscus sinensis , with an average weight of 5.55 g, were fed diets supplemented with eight levels of ferrous sulphate for 8 weeks. The analysed iron content ranged from 50.8 to 482.9 mg kg⁻¹. Growth rate of turtles fed the control diet with no iron supplementation was the lowest among all dietary groups. Haematological parameters including red blood cell, haemoglobin, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration of the turtles fed the control diet were also significantly ( P  < 0.05) lower relative to the other groups. Thus, dietary iron at 50.8 mg kg⁻¹ (no supplemented iron) was deemed deficient for growth and ineffective at preventing anaemia in juvenile soft-shelled turtle. Whereas, a supplementation of 50 mg kg⁻¹ ferrous sulphate (a total dietary iron of 91.8 mg kg⁻¹) was enough to normalize the haematological values of soft-shelled turtles to the level similar to other iron supplement-fed groups. Within the tested dietary iron range, liver iron content curve-linearly ( r ² = 0.99) increased with increasing dietary iron level. Furthermore, thiobarbituric acid-reactive substances in liver tissues of the turtles have also increased when liver iron content increased. The dietary iron requirement of soft-shelled turtle is 120–198 mg kg⁻¹ when ferrous sulphate is used as the source of iron.     

Lidocaine hydrochloride—sodium bicarbonate as an anesthetic for soft-shelled turtle Pelodiscus sinensis

ABSTRACT:   Attempts were made to understand how the different sizes (mean body weight of 4.1 ± 0.8 g for small and 182.6 ± 23.7 g for large) of the soft-shelled turtle Pelodiscus sinensis are affected by different temperature (25°C or 30°C), and different concentrations (700, 1000 and 1300 p.p.m.) of anesthetic lidocaine hydrochloride–sodium bicarbonate. The soft-shelled turtles were fed to satiation once daily. Three-way anova design (size × temperature × concentration of lidocaine hydrochloride) with duplicate was used. Twenty each of large and small soft-shelled turtles were distributed into six 30-L tanks. Exposure time of the soft-shelled turtle was affected by all factors (temperature, concentration and size). Exposure time of the soft-shelled turtle for anesthetizing decreased with increase in temperature and in concentration of lidocaine hydrochloride, and decrease in size. Recovery time for the soft-shelled turtle was also affected by all factors. Recovery time of the soft-shelled turtle increased with increase in temperature, concentration of lidocaine hydrochloride, and size. According to these results, lidocaine hydrochloride (1000 p.p.m.)–sodium bicarbonate seemed an effective anesthetic for sedating and handling the soft-shelled turtle. However, when an anesthetic is recommended for wide use, especially on commercially important edible organisms, the time organism takes to metabolize and excrete anesthetic chemicals from its body to a safe level must be given to allow fitness for human consumption.     

冬眠中华鳖雌、雄生殖道的精子储存

应用光镜和电镜技术,系统观察冬眠中华鳖(Trionyx sinensis)雌、雄生殖道的精子储存情况,显示与精子储存相关的形态结构和细胞特征。结果表明,中华鳖在冬眠期生精活动处于相对静止状态,仅有1～3层精原细胞排列在睾丸曲细精管基膜附近,其余生精细胞排列松散而紊乱,生精上皮中未见有各级细胞规律性排列,不能形成完整的管壁。附睾管腔增大,腔中储存有大量精子。附睾上皮及附睾管腔中见有PAS反应阳性物质。附睾上皮主要由主细胞、亮细胞和基细胞组成。超微结构显示,附睾上皮细胞中含有大量分泌颗粒,内质网膨大。储存在附睾中的精子结构完整,精子中段由35～40个同心圆状线粒体构成线粒体鞘,中段胞质中含有大量糖原颗粒。雌性输卵管分布着数量不等的精子,尤其狭部最多。狭部黏膜皱襞高度融合,形成大量纵行于输卵管的储精细管。在细管中含有大量精子,精子头部嵌入上皮细胞纤毛中。蛋白部和子宫部也有少量精子存在,但在这些部位不能形成明显的储精细管。PAS反应显示,输卵管狭部储精细管上皮分泌有糖蛋白类物质。储精细管上皮下固有层中分布有大量的管状腺,通过腺导管与输卵管管腔相通。雌、雄中华鳖隔离4个月(12月初至来年3月底)后,在雌性输卵管峡部的储精细管中仍可观察到大量结构完整的精子,表明中华鳖精子在输卵管中至少可以储存120 d。以上结果显示,在冬眠期,雄性附睾和雌性输卵管中有大量精子储存,储存的精子能够渡过漫长的冬季用于来年春天交配或受精,这一特殊的生殖策略对其成功繁殖具有重要意义。本研究还对中华鳖的繁殖特性进行了讨论。     

Effects of stocking density on growth and non-specific immune responses in juvenile soft-shelled turtle, Pelodiscus sinensis

To investigate the influence of stocking density on the growth and non‐specific immune responses of juvenile soft‐shelled turtle, Pelodiscus sinensis, three groups of turtles (initial body weight 74.2±12.2 g) were reared in 80 cm (L) × 40 cm (W) × 30 cm (H) tanks for 35 days at three stocking densities (StD), which were 1 individuals (ind.) tank⁻¹ (3.13 ind m⁻²) for D1, 4 ind. tank⁻¹ (12.5 ind m⁻²) for D2 and 6 ind. tank⁻¹ (18.75 ind m⁻²) for D3 respectively. D1 was the control group. Turtles were weighted individually at the beginning and at the end of the trial. At the end of the rearing trial, non‐specific immune parameters were determined. The results showed that the heterophil:lymphocyte ratio (HLR) increased and the total plasma protein decreased with increasing StD, suggesting a stress response. Specific growth rate (SGR) significantly decreased with increasing StD. Non‐specific immune indicators such as blood cell phagocytic activity, serum haemolytic activity and serum bacteriolytic activity increased clearly at a higher StD, indicating a pattern of immunoenhancement. The results indicated that at StDs of 4 and 6 ind. tank⁻¹, non‐specific immune responses were not suppressed compared with the control, but were inversely enhanced at the cost of growth reduction. A possible energy‐based trade‐off between growth and constitutive immunity may exist in soft‐shelled turtles.     

Effect of dietary calcium and phosphorus supplementation in fish meal-based diets on the growth of soft-shelled turtle Pelodiscus sinensis (Wiegmann)

A growth experiment was conducted to determine the effect of supplementing dietary calcium in fish meal‐based diets on the growth of cultured soft‐shelled turtle Pelodiscus sinensis. Juvenile soft‐shelled turtles of 4.1 g mean body weight were fed nine diets containing two levels of phosphorus (2.7% or 3.0%) and analysed calcium levels ranging from 4.7% to 6.6% for 10 weeks. The growth of the turtles was enhanced when inorganic calcium was added to the diets. The weight gain of the turtles fed the control diet containing calcium solely from fish meal was the lowest among the test groups, and was significantly lower than those fed the diet containing 5.7% calcium at the 3.0% phosphorus level (P<0.05). Feed conversion and protein efficiency ratios were not affected by different dietary treatments. Whole‐body moisture and crude protein contents of turtles were not affected by different dietary treatments. The body ash of turtles fed 3.0% phosphorus diets tended to be higher than turtles fed 2.7% phosphorus diets. The body calcium to phosphorus ratio of turtles fed 3.0% phosphorus diets was greater than that of turtles fed diets containing 2.7% phosphorus. Supplementation of Ca in a fish meal‐based practical diet is required for the optimum growth of soft‐shelled turtles.     

中华鳖生态养殖期的死亡率及其在不同池塘环境下的差异

对5种类型养殖池塘90400只中华鳖在生态养殖期前100d的死亡率进行研究。结果说明,中华鳖温室转外塘生态养殖期的日死亡率呈对数方程分布,且可分成3个养殖阶段,分别为日死亡率快速下降期,日死亡率稳步下降期和日死亡率无显著变化期。中华鳖的日死亡率在不同池塘环境下有显著性的差异,表现为土质池塘的死亡率明显低于水泥池塘,在养殖池塘中种植一定面积的水生植物亦能减低中华鳖的死亡率。总体来说,在其他条件相同的情况下,中华鳖最佳的生存环境为土质池塘种植30％～50％的沉水型水生植物,并且保持池塘生态环境处于相对稳定。     

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).     

中华鳖趋化因子CXCL10基因的组织分布、对细菌的响应及其原核表达分析

为研究趋化因子CXCL10的表达特征及其对外源病原微生物的响应,该研究以中华鳖为实验对象,利用生物信息学分析CXCL10蛋白的分子特征及系统进化,通过荧光定量PCR技术检测该基因在健康中华鳖各组织中的表达分布及其在革兰氏阳性菌金黄色葡萄球菌(Staphylococcus aureus)和革兰氏阴性菌嗜水气单胞菌(Aeromonas hydrophila)胁迫下mRNA的表达规律。结果显示该蛋白分子量为10.89 ku,等电点为9.62,存在信号肽和磷酸化位点,单链包含1个N端的loop区、1个α螺旋和3个反向平行β折叠;该基因在健康中华鳖的心脏和肝脏中表达量最高,在血液中表达量最低;两种细菌的刺激能促使中华鳖组织器官中CXCL10及炎症相关因子TNF-ɑ、IL-1βmRNA表达量的显著上调,表明CXCL10基因能够参与中华鳖机体抗菌免疫过程。同时本实验构建pET-32a-CXCL10重组质粒,通过原核表达系统体外得到分子量约为29.7 ku的CXCL10融合蛋白,为进一步探究该蛋白的体外抑菌功能奠定基础。     

利用正交实验法对中华鳖免疫增强剂的初步研究

利用L9（3^4）正交表,通过正交实验设计初步研究了免疫多糖、低聚异麦芽糖和茶花粉等3种免疫增强剂对中华鳖免疫功能的影响。实验结果表明：以存活率为考察指标,影响程度排序为低聚异麦芽糖〉免疫多糖〉茶花粉,低聚异麦芽糖最适水平为0.3%;免疫多糖最适添加水平为0.1%或0.3%,两水平间无显著差异;不同水平的茶花粉对中华鳖存活率影响均不显著。以脾脏指数为考察指标,影响程度排序为低聚异麦芽糖〉茶花粉〉免疫多糖,但3种免疫增强剂的影响程度不显著;同样,以溶菌酶为考察指标,影响程度排序为茶花粉〉免疫多糖〉低聚异麦芽糖,但3种免疫增强剂的影响程度不显著。因此低聚异麦芽糖和免疫多糖比较适合用作中华鳖的免疫增强剂。