Correlation of hematology and plasma chemistry levels in silver crucian carp Carassius langsdorfii

ABSTRACT: The correlation between hematology and plasma chemistry was examined in natural silver crucian carp Carassius langsdorfii. Three microsatellite DNA markers identified five clonal lines. Blood was obtained from the caudal vessels and analyzed using automatic analysis systems. The correlation between several parameters including erythrocyte count (RBC), mean corpuscular hemoglobin concentration (MCHC), alkaline phosphatase (ALP), plasma glucose (Glu) and total protein (TP) was assessed by liner regression. Hemoglobin (Hgb) and hematocrit (Hct) as a function of the erythrocyte count, hematocrit as a function of the hemoglobin, mean corpuscular hemoglobin (MCH) as a function of the mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) as a function of the mean corpuscular hemoglobin, and total protein as a function of triglyceride (TG) were positively correlated for the five clonal lines. Different regression slopes ( b -value) of RBC to Hgb and Hct were found among clonal lines, while no such differences were found among slopes of the remaining regression equation. This phenomenon suggests that the significantly different slopes among parameters, which indicated low genetic influence at the each parameter, clearly detected the genetic influences as the compound effect of these parameters.     

Genetic and environmental variances on growth and reproductive traits of silver crucian carp Carassius langsdorfii using communal and separate rearing systems

SUMMARY: Five clonal lines of silver crucian carp Carassius langsdorfii were produced from natural maternal fish to estimate the variance and heritability of total length, bodyweight and gonadosomatic index (GSI). The larvae on the 10th day after hatching were reared together in a common rearing system with five duplicate systems. Another system was prepared using a separate system with two replications in each family. Estimation of variance and heritability were done in the first, third, fifth and 10th month for total length and bodyweight. Microsatellite DNA markers were used to identify the clonal lines in each communal rearing tank. Average heritabilities in the first month were 0.105 for total length (ranging from 0.21 to 0.03) and 0.127 for bodyweight (ranging from 0.22 to 0.06) then decreased in the third, fifth and 10th month. In contrast, intermediate levels of heritability from separate systems were observed. Likewise, relative heritability of GSI in the 10th month presented a high level in both rearing systems. Those heritabilities indicated that growth, total length, bodyweight and reproductive performances of silver crucian carp were affected by genetic influence. The results suggest that clonal lines of silver crucian carp are suitable as experimental animals for evaluations of environmental change and various pollutants through quantitative traits such as growth, reproductive and other physiological traits.     

银鲫与普通鲫群体遗传结构的比较分析

利用15个微卫星多态性分子标记,对黑龙江省方正县双龙水库和双凤水库两个水体的雌核发育银鲫(Car-assius auratus gibelio)和二倍体普通鲫(Carassius auratus(L))的群体遗传多样性进行比较分析。结果显示:平均每个多态性位点获得银鲫10.39个有效等位基因,普通鲫11.82个有效等位基因;双龙水库三倍体银鲫和二倍体普通鲫的平均观测杂合度分别为0.6035和0.6300,双凤水库中的三倍体银鲫和二倍体普通鲫的平均观测杂合度为0.5311和0.5885;相似性与遗传距离分析结果显示:相似性最大的是双凤水体内的银鲫和普通鲫(0.8127),远大于两个水体间的银鲫群体间的相似性。结果表明,银鲫与普通鲫存在着明显的遗传物质交流,较高的遗传相似性不仅与传统的银鲫起源于普通鲫的观点一致,也暗示银鲫与普通鲫存在经常性地遗传物质交流。     

雄核发育异育银鲫及其初步应用

用鲫（２ｎ＝１００）卵与异育银鲫（３ｎ＝１５６）正常精子进行人工授精,产生了部分的雄核发育异育银鲫（３ｎ＝１５６）。池塘中饲养的雄核发育能银鲫性腺不仅能成熟,而且也能生殖。它与父本异育银鲫回交可以产生回交子一代（Ｂ１）。雄核发育异育银鲫、回交子一代异育银鲫与父本异育银鲫非常相似。它们均具有相同的染色体数目（３ｎ＝１５６）,红细胞核的体积是鲫红细胞核体积的１．６倍。另一方面,用随机拉增多态ＤＮＡ（Ｒ     

彭泽鲫的分子遗传分析及其与方正银鲫A系的比较

Genetic homogeneity between Pengze crucian carp and strain A of silver crucian carp was studied by using transferrin,isozyme and RAPD markers.The studied individuals of Pengze crucian carp showed transferring patterns were the same of silver crucian carp A strain while distinct from those of other crucian carp populations.As far as isozyme is concerned,the MDH,LDH and EST are all of the same with only slight differences in SOD between them.The RPAD patterns clearly indicated high homogeneity among 16 individuals (6 sampled from individuals of two years old and the others aged one) from crucian carp of Pengze and 5 individuals from strain A of silver crucain carp.Nearly indentical banding patterns were observed among all individuals.Average genetic distance within all the individuals is only 0.011,suggesting crucian carp of Pengze might possess indentical genetic background with strain A of silver crucian carp.     

壳聚糖对异育银鲫生长和消化酶的影响

以基础饲料为对照,以在基础饲料中分别添加0.125%、0.25%、0.5%、0.75%、1%的壳聚糖为实验饲料,饲养异育银鲫（Carassius auratus gibelio）60 d,与对照组比较生长、成活、饲料系数、蛋白质效率、鱼体营养成分、白肌RNA/DNA比值及肝胰脏、肠道的蛋白酶、淀粉酶活性.结果表明,本次实验所添加壳聚糖对异育银鲫的成活率、鱼体营养成分和白肌的RNA/DNA比值均无显著影响（P＞0.05）.添加0.5%、0.75%和1%壳聚糖可以显著提高异育银鲫的特定生长率和蛋白质效率,显著降低异育银鲫的饲料系数.添加0.5%壳聚糖可以显著提高异育银鲫肠道的淀粉酶活性;添加0.75%和1%壳聚糖可以显著提高肠道和肝胰脏的蛋白酶活性和肠道的淀粉酶活性.[中国水产科学,2006,13（3）：440-445]     

异育银鲫实验性免疫抑制模型的建立

分别以每kg体重鱼注射0、1、10、20mg环磷酰胺的剂量、每隔5d给异育银鲫注射1次,共注射3次,第16天检测各组试验鱼外周血白细胞数量及吞噬活性、不同类型的白细胞数量变化、脾脏和头肾的溶菌酶活性、体外培养外周血B淋巴细胞分泌的IgM、IFN-α含量、外周血NK细胞杀伤功能及头肾、脾脏、外周血B淋巴细胞转化能力。结果表明,环磷酰胺剂量的增加可以使试验鱼外周血白细胞的数量包括各种类型的白细胞的数量、体外培养外周血B淋巴细胞分泌的IgM、IFN-α含量下降,脾脏和头肾的溶菌酶活性、外周血白细胞吞噬活性及头肾、脾脏、外周血B淋巴细胞转化能力降低。提出每kg鱼10mg的环磷酰胺是建立异育银鲫实验性免疫抑制模型的适宜剂量     

鲫肠道上皮细胞原代培养方法的研究

探讨了健康鲫(Carassius auratus)幼鱼肠道上皮细胞的原代培养方法。结果显示,用含有抗生素、胶原蛋白酶Ⅰ、EDTA和胶原蛋白酶Ⅳ组成的D-Hanks液消化无菌分离的鲫肠道可以获得大量细胞团及绒毛隐窝;用含有抗生素、胎牛血清(FBS)、表皮生长因子(EGF)、胰岛素的DMEM培养液进行培养,并根据肠道上皮细胞与成纤维细胞贴壁时间的差异进一步纯化,连续培养12d后可以得到纯化的肠道上皮细胞。     

用生长和非特异性免疫力评定异育银鲫维生素C需要量

用维生素 C(Vc)含量为0～1200mg·kg~(-1)的精制饲料,连续投喂异育银鲫8周后,测定异育银鲫生长速度和非特异性免疫指标。结果表明,在饲料Vc含量达150mg·kg~(-1)时日增重率最大,进一步提高饲料Vc含量,日增重率没有显著变化;饲料Vc含量为75mg·kg~(-1)时头肾指数显著提高,而脾指数不受饲料Vc含量的影响;头肾细胞吞噬活性和A花环率均在Vc含量为300mg·kg~(-1)时达最高水平;而血清溶菌酶活性和总补体活性分别在饲料Vc为300mg·kg~(-1)和600mg·kg~(-1)时达最高值。上述结果表明饲料Vc可显著提高异育银鲫免疫力。鉴于Vc的免疫促进作用,建议在疾病多发季节饲料Vc添加量提高到300mg·kg~(-1)。     

饲料Vc对异育银鲫的生理效应及其适宜添加量

在精制饲料中分别添加不同剂量Vc(0、37.5、75、15 0、30 0、6 0 0和 12 0 0mg/kg) ,连续投喂异育银鲫 (方正银鲫♀×兴国红鲤♂ ) (Carassiusauratusgitelio) 8周。结果显示 ,当饲料中缺乏Vc时异育银鲫虽没有出现明显的缺乏症状 ,但日增重率、血红蛋白含量和血清POD酶活性均低于Vc添加组 ,而红细胞脆性极显著地高于高剂量Vc添加组 (P <0 .0 1) ;同时 ,经新复极差检验 ,在适宜范围内随着饲料Vc含量增加 ,异育银鲫日增重率、血红蛋白含量、POD活性等指标显著提高 ,而红细胞脆性明显下降 (P <0 .0 5 )。综合以上指标的非线性回归分析结果 ,建议生产上异育银鲫饲料Vc适宜添加量为 30 0～ 5 0 0mg/kg。     

饲料糖种类和水平对异育银鲫肝糖原代谢的影响

用分别含25％葡萄糖、25％淀粉及不同水平糊精（5％，25％，50％）的5种饲料喂养异育银鲫（方正银鲫♀X兴国红鲤♂）6周，然后禁食1、2、4周，以摄食后血糖和肝糖原的变化规律及禁食反应来研究肝糖原代谢。实验钍体重约15g。结果表明，摄食后25％葡萄糖组血糖升高速度最快，升幅最大，5％糊精组血糖最低。各组鱼的肝糖原含量在摄食后4h左右达到最大值，但葡萄糖组在摄食后肝糖原含量反而下降。饲养6周后25％葡萄糖组和5％糊精组基础肝糖原含量显著低于其他各组。禁食时25％葡萄糖组和5％糊精组的甘油三酯含量比其他各组低，而总氨基酸水平比其他各组高。上述结果提示在摄食条件下肝糖原的代谢与饲料糖的种类和水平有关，而禁食反应与体内能量物质的储量有关。     

异育银卿口服葡萄糖后血糖、血脂和肝糖原的变化

禁食 4周后对体重为 46± 5g的异育银鲫进行了葡萄糖耐量试验。口服 16 7mg·(10 0g) -1体重的葡萄糖之前及之后的 1、2、3、4、5、6、8、10、14、18和 2 4h分别抽血、取肝胰腺 ,分析血糖、血脂和肝糖原含量。实验结果表明 ,血糖在口服葡萄糖后 3h内迅速升高 ,并于 3h时达最高水平 ,以后逐渐下降。口服葡萄糖后 1h血脂含量最高 ,然后开始下降 ,并于 8h时又开始升高。口服葡萄糖后的最初 2h内肝糖原含量显著降低 ,随后开始提高 ,并在 6h时达最高峰。用体重为 (16 4± 12 )g的异育银鲫为材料禁食 4周后灌喂不同剂量的葡萄糖 ,研究葡萄糖剂量对血糖和肝糖原变化规律的影响。结果表明 ,随着口服葡萄糖剂量的增加 ,血糖升高幅度加大 ,而肝糖原含量则随葡萄糖剂量增加而减少。上述实验结果提示异育银鲫在胰岛素的分泌方面与哺乳类存在本质的不同。     

多氯联苯(PCB1254)对异育银鲫血清和离体卵巢分泌睾酮和雌二醇的影响

采用活体注射和卵巢体外培养添加多氯联苯（PCB₁₂₅₄）两种方式研究了PCB₁₂₅₄对异育银鲫血清中和离体卵巢分泌睾酮及雌二醇的影响。注射PCB₁₂₅₄后，血清中睾酮浓度随作用时间延长而降低，随PCB₁₂₅₄浓度的增加而减小；血清中雌二醇随作用时间延长而增加,随PCB₁₂₅₄浓度增加而增加。体外培养时卵巢时，培养液中雌二醇含量随培养时间延长而增加，随PCB₁₂₅₄浓度增加而增加。结果表明PCB₁₂₅₄促进雌二醇分泌，抑 制睾酮分泌。     

饲料中添加芽孢杆菌和硒酵母对异育银鲫的生长及抗病力的影响

在基础饲料中分别添加0.1%、0.2%、0.4%的芽孢杆菌,0.3%、0.6%、1.2%的硒酵母,0.2%+0.3%、0.2%+0.6%、0.2%+1.2%的芽孢杆菌和硒酵母混合物,投喂异育银鲫2个月.测定鱼体的生长和对病原菌嗜水气单胞菌的抗感染能力--LD50.结果表明,添加不同浓度的芽孢杆菌对体重的增加有显著影响(P＜0.05),其中添加0.2%的芽孢杆菌组相对增重率比对照组增加33%.添加不同浓度的硒酵母均可不同程度地促进异育银鲫的生长(P＜0.05),其相对增重率比对照组增加16%～31%.不同配比的芽孢杆菌和硒酵母混合物显著性提高鱼的相对增重率,但增重效果与单一添加芽孢杆菌或硒酵母相似(P＞0.05).饲料中单一添加芽孢杆菌或硒酵母可以极显著地提高异育银鲫对嗜水气单胞菌的抵抗能力,添加效果在各芽孢杆菌水平组间无显著差异,随硒酵母添加量的增大而增大(P＜0.01),但0.6%组和1.2%组无显著差异.混合添加芽孢杆菌和硒酵母也能极显著增强异育银鲫对嗜水气单胞菌的抗感染能力(P＜0.01),但添加效果不及单一芽孢杆菌;与单一硒酵母相比,抗感染能力与硒酵母的添加量有关(P＜0.01),添加量增大,抗感染能力减弱(P＜0.05或P＜0.01).     

壳聚糖和益生菌对异育银鲫非特异免疫功能及血清甲状腺激素、皮质醇水平的影响

为探讨壳聚糖和益生菌是否通过甲状腺激素、皮质醇对异育银鲫的非特异免疫功能的进行调控,试验以基础饲料为对照,在基础饲料中分别添加0.50%壳聚糖A、0.50%壳聚糖B、0.20%益生菌以及0.50%壳聚糖A与0.05%益生菌的混合物制备试验饲料,饲养异育银鲫64d后,测定各组鱼外周血白细胞数量及其分类、白细胞吞噬活性、脾脏溶菌酶活性、脾脏淋巴细胞转化能力;同时采用放射免疫分析法测定异育银鲫血清中三碘甲腺原氨酸(T3)、甲状腺素(T4)及皮质醇含量。结果表明,饲料中添加壳聚糖、益生菌或其混合物对异育银鲫白细胞总数无显著影响,但是可显著提高异育银鲫外周血淋巴细胞数量、白细胞吞噬活性和脾脏溶菌酶活性(P<0.05),显著降低外周血多形核细胞数量(P<0.05)。除了壳聚糖A能够显著提高异育银鲫脾脏B淋巴细胞转化能力之外,其余试验组的脾脏T、B淋巴细胞转化能力均与对照组无显著差异(P>0.05)。同时发现饲料中添加壳聚糖、益生菌或其混合物可以显著提高异育银鲫血清中T3的含量(P<0.05)、显著降低异育银鲫血清中皮质醇的含量(P<0.05),除了壳聚糖B对异育银鲫血清中T4的含量无显著影响(P>0.05)之外,其余均可...     

Effects of a dietary administration of purple coneflower (Echinacea purpurea) on growth,antioxidant activities and 8 miRNAs expressions in crucian carp (Carassius auratus)

Echinacea purpurea (EP), a globally popular herbal medicine, has been used to treat various diseases in human and animals. However, little has been reported about its effects in fish. In this study, crucian carp (Carassius auratus) were selected to evaluate the effects of EP on growth performance and antioxidant response and the expressions of microRNAs. The results showed EP could stimulate the growth of crucian carp with the best effect was observed at dose of 4 g kg^?1. In serum, the content of hydroxyl radical (·OH) and malondialdehyde (MDA) decreased by EP supplementation, whereas the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) increased. Similarly, the content of ·OH decreased, and the activities of CAT, GPX and GR increased in liver of crucian carp. Furthermore, in livers of crucian carp, EP supplements upregulated the expressions of the microRNAs (miR‐16, miR‐21, miR‐125b, miR‐146a, miR‐155, miR‐181a and miR‐223), which had been confirmed to participate in regulating antioxidant and immune function in mammals. Our results suggest EP supplements in diets stimulated growth performance and antioxidant response of crucian carp. In liver, the upregulation of specific miRNAs may participate in the antioxidant effects of EP diets.     

Effect of soybean meal,raffinose and stachyose on the growth,body composition,intestinal morphology and intestinal microflora of juvenile allogynogenetic silver crucian carp (Carassius auratus gibelio♀×Cyprinus carpio♂)

The study was conducted to investigate the effects of soybean meal (SBM), raffinose and stachyose on juvenile crucian carp (Carassius auratus gibelio♀ × Cyprinus carpio♂). The experimental diets consisted of one control diet based on fish meal (FM), one diet containing 300 g kg^?1 SBM and four FM‐based diets with the addition of either 6.7 g kg^?1 raffinose (Raf), 33.9 g kg^?1 stachyose (Sta), a combination of raffinose and stachyose (Raf?Sta) and finally a Raf?Sta diet supplemented with 2.5 g kg^?1 saponins (Raf?Sta?Sap). After 3 weeks of feeding, the relative gut lengths of SBM‐fed fish and the fish fed stachyose‐containing diets were shorter than those of the FM‐fed fish; further, more SBM‐fed fish showed fissures on the tips of the intestinal folds. After 8 weeks of feeding, the growth of SBM‐fed fish was significantly lower than that of FM‐fed fish (P<0.05). The fish fed Raf?Sta?Sap had a low relative gut length (P<0.05). In comparison with the other fish, the SBM‐fed fish had a higher number of large‐sized homogeneous vacuoles in the cytoplasm of epithelial cells and shorter microvilli. No significant difference was observed in body composition or intestinal microflora. The results indicated that raffinose and stachyose played no or only minor roles in the development of soybean‐induced growth reduction.     

额尔齐斯河2种类型雌性银鲫的形态特征及D-loop基因序列比较

2008年5月调查发现,在额尔齐斯河主河道(185团)的雌性银鲫(Carassius auratus gibelio)群体中存在2种形态类型,按照体长比体高的形态学性状区分,分别为高背型(1.78~2.14)和低背型(2.18~2.39)。形态学分析表明,高背型和低背型的侧线鳞分别为31和30,侧线上(下)鳞为分别7和6,两者在体高、头高、尾柄高、体宽、头宽等性状上存在显著差异(P<0.01),高背型比低背型具有明显的体型优势。生长特性分析表明,在相同体长或年龄的雌性群体中,高背型的平均体质量约为低背型的1.3~1.9倍,而表现出更为优越的生长性能。线粒体控制区序列分析表明,高背型和低背型群体各自独享1个单倍型(分别为EG1和ED1),为2个不同的遗传谱系类群,遗传分化仅为0.004,远低于属内种间的0.020~0.045,两者尚属群体内变异;在分子系统关系中,高背型和低背型分别与方正银鲫的2个单倍型(FZ1和FZ13)共享并聚为单系,推测额尔齐斯河银鲫源自于黑龙江(方正银鲫)。与国内其他地方群体或品种比较表明,额尔齐斯河2种类型雌性银鲫,特别是高背型具有更为突出的生长优势,可作为优良的开发养殖品种和更具潜力的鱼类育种材料。     

银鲫肠道内抑制嗜水气单胞菌的潜在益生菌筛选及其特性研究

从养殖的银鲫(Carassius auratus gibelio)肠道中分离纯化得到60株具有不同特征的菌株,以嗜水气单胞菌(Aeromonas hydrophila)为指示菌,点种法初筛得到12株菌株,再通过牛津杯实验筛选出4株对嗜水气单胞菌有明显抑制作用(抑菌圈直径在14 mm以上)的益生菌疑似株,并进行了耐热试验、耐酸试验和人工肠液耐受试验,筛选出最符合益生菌功能特性的菌株YJ-9。该菌株在90℃高温水浴30 min后相对存活率为1.51%,在pH3的液体培养基中4 h内相对存活率为8.37%,活菌数都保持在106 CFU/mL以上,并且可以在人工模拟肠液中缓慢生长。在急性毒力实验中,10倍于实验剂量的菌株YJ-9对银鲫无毒,动物保护实验证明菌株YJ-9能有效增强银鲫对嗜水气单胞菌的免疫抵抗力,相对保护率达到80%。研究结果表明,菌株YJ-9可以作为一个新的饲用微生态制剂候选菌种。