虾青素对微囊藻毒素MC-LR胁迫下克氏原螯虾的生长、繁殖及免疫力的影响

本文目的旨在研究虾青素对微囊藻毒素MC-LR胁迫下(25μg/L)克氏原螯虾的免疫力及生长繁殖的影响。实验设计四个梯度的饲料中虾青素添加比例分别为(0、5、10、20 mg/g)养殖周期持续8星期。结果表明虾青素显著提高了克氏原螯虾肝脏中超氧化物歧化酶(SOD)和碱性磷酸(AKP)的活力(P0.05)。5g/kg虾青素的添加量即可对SOD酶活具有明显提升(P0.05);10 g/kg的虾青素添加量对AKP的提升最为明显(P0.05);研究还发现虾青素可显著提高微囊藻毒素MC-LR胁迫下克氏原螯虾的存活率和特定生长率(SGR)(P0.05),提高克氏原螯虾雌虾的抱卵量。研究表明虾青素作为一种抗氧化剂适宜于在克氏原螯虾饲料中添加,并且具有提高虾体免疫活力,缓解微囊藻毒素MC-LR对虾体氧化胁迫的作用。     

细菌糖蛋白对螯虾免疫因子的影响

实验检测了气味黄杆菌的胞外糖蛋白对克氏原螯虾（Ｐｒｏｃａｍｂａｒｕｓ ｃｌａｒｋｉｉ）的几咱非特异性免疫因子－凝集素、酚氧化酶、溶菌、抗菌和超氧化物歧化酶等活力的影响。注射１０ｍｇ糖蛋白后,实验组敖蛳主血清的免疫因子活力均高于对照组,凝集素活力在第１～４天最高;酚氧化酶在第１～５天维持较高活力,最大活力在第２天,溶菌、抗菌活力在第１～６天维持较高水平,分别在第３天、第２天达到最大。与对照组相比,实     

Effects of ammonia exposure on stress and immune response in juvenile turbot (Scophthalmus maximus)

The aim of this study was to investigate the effects of ammonia exposure on stress and immune response in turbot. The species was exposed to five total ammonia nitrogen (TAN) concentrations: 0 (control), 1, 5, 20 and 40 mg L^?1 for 96 h. After 0, 24, 48 and 96 h of exposure, blood samples were collected to measure the levels of corticotropin‐releasing hormone (CRH), adrenocorticotropic hormone (ACTH), cortisol, growth hormone (GH), lysozyme (LZM), complement 3 (C3), complement 4 (C4) and immunoglobulinM (IgM); liver samples were taken to analyse oxidative stress parameters (superoxide dismutase, SOD; catalase, CAT; glutathione, GSH; malondialdehyde, MDA), and gene expression of heat shock proteins (HSP 70 and HSP 90) and insulin‐like growth factor‐1 (IGF‐1). The results showed that exposure to higher concentrations of TAN (20 and/or 40 mg L^?1) enhanced the levels of CRH, ACTH and cortisol and attenuated the levels of GH, LZM, C3, C4 and IgM in plasma of turbot after 48 and 96 h. In liver, TAN (20 and/or 40 mg L^?1) apparently increased the activities of antioxidative enzymes (SOD and CAT), mRNA levels of HSP (HSP 70 and 90) and formation of MDA, decreased the content of GSH and mRNA levels of IGF‐1 after 48 and 96 h of exposure. Overall, our results suggested that high ammonia exposure caused activation of hypothalamic–pituitary–interrenal axis, inhibition of GH/IGF axis and immunity, and occurrence of oxidative stress.     

克氏原螯虾PcCRCN-L基因的克隆鉴定及在低氧胁迫下的表达响应

虾青蛋白(Crustacyanin,CRCN)在甲壳动物脂类代谢及低氧胁迫应激调控等方面具有重要的功能。为获取虾青蛋白在克氏原螯虾(Procambarus clarkii)性腺发育和低氧胁迫应答中的作用,本研究在克氏原螯虾肝胰腺组织中鉴定出1个类虾青蛋白PcCRCN-L基因的c DNA序列,分析了PcCRCN-L基因的结构特征和进化模式,研究了PcCRCN-L基因在不同组织与性腺不同发育时期的表达特征,探讨了Pc CRCN-L在低氧–复氧胁迫下的表达响应模式。结果显示,PcCRCN-L基因c DNA序列长2 700 bp,其开放阅读框(ORF)长度为1 587 bp,编码528个氨基酸残基;DNA序列长6 130 bp,位于克氏原螯虾基因组的第12号染色体,包含5个外显子和4个内含子,内含子/外显子剪接方式符合GT-AG规则。Pc CRCN-L具有1个完整的lipocalin结构域,包含典型的序列保守区Ⅰ(SCR1)序列G-X-W、保守区Ⅱ(SCR2)序列T-D-Y和保守区Ⅲ(SCR3)序列精氨酸R。多序列比对与系统进化分析结果显示,PcCRCN-L独立于传统虾青蛋白A和C亚族,单独聚...     

Effects of ammonia exposure on oxidative stress,immune enzyme activities,and intestinal microbiota of Pacific white shrimp Litopenaeus vannamei

Aquaculture International - An ammonia stress experiment was conducted by adjusting the concentration of total ammonia nitrogen (TAN) to 20 mg/L using ammonium chloride. The shrimp without...     

克氏螯虾染色体研究

本文以克氏螯虾精巢为材料，用改良常规气干法进行染色体制片并显微照相，观察结果表明，克氏螯虾精母细胞染色体数为９４，中期染色体呈颗粒状。     

克氏原螯虾对高温应激的生理学响应

本实验旨在研究高温对克氏原螯虾(Procambarus clarkia)血液生化指标、抗氧化能力以及热休克蛋白70基因表达的影响。实验分别采用30℃和35℃水温进行高温应激试验,应激前(0 h)和在应激后3、6、12、24和48 h分别采集样本,测定血淋巴液中LDH、GLU、TCHO、TP和TG含量,测定肝胰腺T-AOC、CAT、MDA以及HSP70基因的表达变化。结果显示:在高温应激的初级阶段(6 h),克氏原螯虾血淋巴中的TCHO和LD含量急剧上升,而GLU含量持续上升到24 h;TG含量在应激的初级阶段变化不明显,但在6 h后急速上升,在12 h达到最高后开始回落;TP的含量在初级阶段出现下降,然后在12 h恢复至应激前水平。肝胰腺中的MDA含量在应激后持续上升,在24 h后达到最高值,随后下降至应激前水平;T-AOC在应激后3 h上升至最高水平,随后降低;CAT的含量分别在应激后6 h和12 h上升至最高值然后回落。HSP70基因在应激前未检测到有表达,高温应激后3 h即可检测到少量表达,35℃组在6 h出现大量表达,并持续到12 h,随后24 h降低至无法检出;而30℃组在应激6 h后降低至无法检出。结果表明,30℃和35℃高温能引起克氏原螯虾机体产生应激反应,并对机体的生理机能产生影响。     

克氏原螯虾繁殖生物学研究

对克氏原螯虾(Procambarus clarkii)繁殖生物学进行了研究。结果显示:克氏原螯虾的性成熟年龄为1年左右;雌虾最小体长为6.4 cm,最小体重为10 g;雄虾最小体长为7.1 cm,最小体重为20 g。5~9月为交配期,其中以6~8月为交配高峰期。克氏原螯虾交配后大约为30 d左右产卵。繁殖期为7~10月,高峰期为8~9月。10月底以后抱卵的虾由于水温逐步降低,一直延续到第2年春季才孵化。克氏原螯虾的繁殖行为与掘洞行为密切相关,繁殖期的掘洞数量较非繁殖期明显增多。7~10月,亲虾均栖息在洞内繁殖,洞穴深度为50~80 cm。卵巢为一次产卵类型。雌雄比例为1∶1。克氏原螯虾的个体绝对繁殖力的变动范围为172~1158粒,平均为517粒。个体相对繁殖力(F/W)的变动范围为2~41粒/g,平均为21粒。个体相对繁殖力(F/L)的变动范围为47~80粒/cm,平均为63粒。     

克氏原螫虾繁殖生物学研究

对克氏原螯虾(Procambarus clarkii)繁殖生物学进行了研究.结果显示:克氏原螯虾的性成熟年龄为1年左右;雌虾最小体长为6.4 cm,最小体重为10 g;雄虾最小体长为7.1 cm,最小体重为20 g.5～9月为交配期,其中以6～8月为交配高峰期.克氏原螯虾交配后大约为30 d左右产卵.繁殖期为7～10月,高峰期为8～9月.10月底以后抱卵的虾由于水温逐步降低,一直延续到第2年春季才孵化.克氏原螯虾的繁殖行为与掘洞行为密切相关,繁殖期的掘洞数量较非繁殖期明显增多.7～10月,亲虾均柄息在洞内繁殖,洞穴深度为50～80 cm.卵巢为一次产卵类型.雌雄比例为1:1.克氏原螯虾的个体绝对繁殖力的变动范围为172～1158粒,平均为517粒.个体相对繁殖力(F/W)的变动范围为2～41粒/g,平均为21粒.个体相对繁殖力(F/L)的变动范围为47～80粒/cm,平均为63粒.     

The Effect of Temperature on Feed Consumption and Nutrient Absorption in Procambarus clarkii and Procambarus zonangulus

Feeding characteristics of acclimated red swamp crayfish Procambarus clarkii and white river crayfish Procambarus zonangulus were determined at 8, 14, 20, 26, and 32 C. Food consumption rates of crayfish were directly related to temperature. Feeding was minimal at 8 C. At 14 CP. zonangulus consumed significantly more feed (5.5% body weight (BW/d) than P. clarkii (1.8% BW/d). At 32 C P. clarkii consumed significantly more feed (23% BW/d) than P. zonangulus (10% BW /d), reflecting species-specific thermal optimums for feeding. Differences in food consumption rates correspond to differences in the geographical distributions of these two species. Dry matter absorption rates ranged from 55% to 75% for both species without significant temperature effect. Patterns of protein and carbohydrate absorption were similar in P. clarkii and P. zonangulus. Protein absorption efficiency decreased from 45% to 10% as temperature increased from 8 C to 32 C. The total amount of protein absorbed increased seven- to ten-fold as temperature increased. Soluble carbohydrate absorption efficiency was reduced at low temperatures. However, absorption efficiency increased approximately five-fold as temperature increased from 14 C to 20 C, but did not change substantially as temperature was increased to 32 C. The total amount of carbohydrate absorbed increased approximately 15- to 26-fold as temperature increased. The protein to carbohydrate utilization ratio was similar between species, but varied dramatically with temperature. Both species absorbed between 2.5 and four-times more protein than carbohydrate at low temperatures (8 C and 14 C). At 32 C both species utilized approximately three times more carbohydrate than protein. In spite of reduced consumption at lower temperatures, crayfish absorbed a greater protein portion. At these low temperatures it appears that crayfish maximize the efficiency of protein absorption, presumably for maintenance and growth requirements. At high temperatures it appears that crayfish maximize energy intake, presumably to sustain higher metabolic rates due to increased activity levels. These apparent temperature-dependent nutrient requirements may influence natural foraging strategies or, for species in aquaculture, the effectiveness of commercial feed preparations. At high temperatures P. clarkii was more effective in consuming nutrients than P. zonangulus and may be more successful than P. zonangulus at these temperatures.     

流式细胞术在克氏原螯虾血细胞的分类、活性和免疫功能研究中的应用

应用流式细胞术(FCM)对克氏原螯虾(Procambarus clarkii)血细胞的分类、活性和免疫功能进行了研究。结果显示:血细胞可分为透明细胞、小颗粒细胞和大颗粒细胞三个亚群,组成比例分别为(26.25±5.29)%、(51.44±7.02)%和(11.20±1.82)%;螯虾血细胞的平均总凋亡率约为3.12%;血细胞对荧光大肠杆菌的吞噬活力显著(P<0.05)高于荧光微球,吞噬率分别为17.04%和14.57%;血细胞在自然生理状态下含有一定量的活性氧,其在两类颗粒细胞的含量显著(P<0.05)高于透明细胞,在大颗粒细胞中最高。结果表明,FCM能较好地应用于虾类的血细胞分类和功能研究。     

克氏原螯虾形态参数关系的初步研究

对594尾(体长1.75~10.6 cm)克氏原螯虾的全长、体长、头胸甲长和体重等进行了测量分析。着重研究了体重与体长的关系,得到的关系式为:W♂=0.0299×Lb3.4925(r2=0.9818),W♀=0.0438×Lb3.1697(r2=0.9869)。同时还推断得出体长6.0~6.5 cm是克氏原螯虾的最适捕捞规格。     

壳聚糖对克氏原螯虾存活、血清生化指标和非特异性免疫功能的影响

以基础饲料为对照,在基础饲料中分别添加0.0%(对照)、0.5%、1.0%、1.5%、2.0%、3.0%的壳聚糖,分别投喂平均体重为(21.55±1.62)g的克氏原螯虾(Procambarus clarkii)60 d,研究壳聚糖对克氏原螯虾存活、蜕壳、血液生化指标及非特异性免疫功能的影响。结果显示:0.5%添加组存活率为84.44%,与对照组和1.0%添加组无显著性差异;总蜕壳率和相对增重率随壳聚糖含量升高而呈现先升高后降低的趋势,当添加量为1.0%时,总蜕壳率和相对增重率显著高于对照组;各试验组肝胰指数差异不显著。2.0%添加组血清中总蛋白、白蛋白和总胆固醇显著高于对照组,添加组甘油三酯的含量显著高于对照组。1.5%添加组血清和肝胰腺中的溶菌酶(LZM)、超氧化物歧化酶(SOD)的活性显著高于对照组;与对照组相比,0.5%添加组血清中的酸性磷酸酶(ACP)和过氧化氢酶(CAT)的活性显著升高;0.5%添加组肝胰腺中CAT活性最高,为4.93 U/mL,显著高于对照组。结果说明,壳聚糖可以提高克氏原螯虾的生长性能和非特异性免疫功能,饲料中添加0.5%～1.5%较为适宜。     

Immunostimulatory effects of dietary fructooligosaccharides on red swamp crayfish,Procambarus clarkii (Girard)

Fructooligosaccharides (FOS) are widely researched as prebiotics and immunostimulants in humans and terrestrial animals. However, limited researches on immune functions of FOS were reported in crustacean. In the present study, red swamp crayfish Procambarus clarkii were fed with different concentrations of FOS (2.0, 5.0, 8.0, 10.0 g kg^?1 diet), and then the immunostimulatory effects of FOS were evaluated by measuring immune parameters and examining immune‐related gene expression. PO and SOD activities were significantly increased in groups fed with 8.0 or 10.0 g kg^?1 of FOS compared with those in control group. In addition, significant elevations of SOD and phagocytic activities were also observed in group fed with 5.0 g kg^?1 of FOS. Among examined immune‐related genes, expressions of crustin1, lysozyme, SOD and pro‐PO were significantly up‐regulated by feeding with 8.0 or 10.0 g kg^?1 of dietary FOS. Survival rates of crayfish challenged with Aeromonas hydrophila were significantly improved by feeding with 5.0, 8.0 or 10.0 g kg^?1 of dietary FOS for 30 days. Phagocytic activities and bacterial clearance efficiencies were also significantly increased in groups fed with 8.0 or 10.0 g kg^?1 of FOS. The results indicated that dietary FOS effectively induced innate immune responses of red swamp crayfish in a dose‐dependent way and could be used as potential immunostimulant in crustacean.     

日粮脂肪水平对克氏原螯虾的生长、体组成及脂肪酸组成的影响

研究了日粮中脂肪水平对克氏原螯虾的生长、体组成及脂肪酸组成的影响。630尾初始重为(4.76±0.42)g克氏原螯虾随机分为六组(1~6组),每组3个重复,每个重复35尾,用六个脂肪水平(1.82%、3.84%、5.79%、7.89%、9.91%和11.95%)饲料饲养60 d后测定虾的生长指标、体组成和脂肪酸组成。结果表明,克氏原螯虾的适宜日粮脂肪水平为7.89%~9.91%。     

淡水龙虾的稻田养殖技术

龙虾学名克氏螯虾，属于节肢动物门。甲壳纲，十足目，龙虾科。目前，全国大多数地区都有龙虾生存，该种属温热带淡水虾类，适应能力强，繁殖率高，食性杂，生长快，抗病，耐高温，耐低氧，离水数小时不会死亡。龙虾肉味鲜美，营养丰富，脂肪含量较低，可食部分达40％以上。鉴于近几年野生龙虾遭到酷捕滥捞，数量急剧下降，为解决这一状况，从2005年开始。我站进行了稻田养殖龙虾试点工作，并取得了一定的成绩。通过实验，我总结了稻田养殖龙虾工作中应注意的几个关键性问题，现就这几个问题将养殖技术简述如下：     

克氏原螯虾掘洞行为研究

克氏原螯虾(Procam barus clarkii)掘洞时间多在夜间,可持续掘洞6~8 h,一夜挖掘深度成虾可达40 cm,幼虾可达25 cm。成虾的洞穴深度大部分在50~80 cm之间,少部分可以达到80~150 cm;幼虾洞穴的深度在10~25 cm之间;体长1.2 cm的稚虾已经具备掘洞的能力,洞穴深度在10~20 cm之间。洞穴分为简单洞穴和复杂洞穴两种:85%的洞穴是简单的,即只有一条穴道,位于水面上、下10 cm之间;15%较复杂,即有2条以上的穴道,位于水面以上20 cm处。洞穴的密度为2.8~5.6个/m2。每个洞穴中一般有1~2只虾,但冬季也常发现一个洞中有3~5只虾。克氏原螯虾在繁殖季节掘洞强度增大,在寒冷的冬季,掘洞强度微弱。     

盐度对克氏原螯虾孵化率的影响

研究了不同盐度对克氏原螯虾受精卵孵化的影响.水温25 ℃条件下,盐度0～8时克氏原螯虾的受精卵均可孵出幼体.但以盐度0～4时孵化率较高,每个亲体平均孵出的幼体数量较多,高出此盐度范围时孵化率明显降低,且随着盐度升高,孵化时间延长.试验表明,0～4的盐度为克氏原螯虾孵化的适宜盐度,孵化率较高,孵化时间较短.     

克氏原螯虾细胞色素c基因通过调节凋亡途径抑制WSSV感染

细胞凋亡是由一系列相关基因严格调控的细胞程序性死亡,在抵御病原入侵、维持机体内环境稳态等方面有着重要意义。其中细胞色素c从线粒体释放到细胞质中是凋亡开始的关键一步。本研究利用RACE技术首次克隆获得了克氏原螯虾(Procambarus clarkii)细胞色素c基因(PcCytc),全长为897 bp,包括163 bp的5′-UTR、419 bp的3′-UTR和315 bp的开放阅读框,编码104个氨基酸。定量PCR检测结果显示,PcCytc基因在克氏原螯虾的各组织中均有表达,其中在鳃、肠道和肌肉中表达高,在胃中表达最低。WSSV感染实验显示,在病毒感染后PcCytc在肝胰腺、肠道和肌肉组织中的表达水平均出现上调,并在24 h达最高值,约是此时PBS组表达量的2.65、2.07和2.20倍,均存在极显著性差异(P<0.01)。PcCytc基因干扰后,克氏原螯虾体内WSSV病毒拷贝数显著增加(P<0.05),表明PcCytc能够抑制WSSV在克氏原螯虾体内的复制,延迟感染;同时,凋亡相关基因bcl-2、bax和caspase-3的表达均发生显著上调或下调(P<0.05)。本研究表明,PcCytc可通过调节凋亡途径抑制WSSV感染,为克氏原螯虾对WSSV感染的免疫反应提供了新的见解。