Cryopreservation of sperm of common carp, Cyprinus carpio L.

Milt of common carp, Cyprinus carpio L. was cryopreserved in pellet form with the use of 12 extenders. Most efficient were: BE2 original extender (containing 85 mM NaCl, 50 mM KCl, 3 mm CaCl₂, 1 mm MgCl₂ with 10% dimethyl-acetamide (DMA) and 10% addition of hen's egg yolk) and Kurokura et al.'s extender with 15% DMA and 10% yolk (about 73% and 69% of eyed eggs, about 61% and 52% of swim-up larvae, respectively). Within the most effective treatments, survival from the eyed-egg stage to the swim-up stage was similar to that observed in the control group. Survival from the eyed-egg stage to the swim-up stage (percentage of eyed eggs was considered as 100%) was highly significantly and positively correlated with the actual rate of swim-up larvae.     

A calcified fibroma in the common carp, Cyprinus carpio L.

Abstract. A large tegumental tumour was observed in a common carp from the pool of the Jardin des Plantes, Montpellier. The tumour was a fibroma characterized by centripetal fibrogenesis and intense calcification of the central region.     

Protection of common carp (Cyprinus carpio L.) spermatozoa motility under oxidative stress by antioxidants and seminal plasma

The protective influence of seminal plasma and the antioxidants catalase (CAT), superoxide dismutase (SOD), and glutathione (GTH) on quality parameters, oxidative stress indices, and antioxidant activity was studied in common carp (Cyprinus carpio) spermatozoa exposed to the xanthine–xanthine oxidase (X–XO) system. Fish spermatozoa were incubated for 5 and 20 min at 4 °C with X–XO concentrations of 1 mM X–0.1 U/mL, 0.6 mM X–0.05 U/mL, 0.3 mM X–0.025 U/mL, and 0.1 mM X–0.0125 U/mL. A dose-dependent reduction in spermatozoa motility and velocity was observed at concentrations of 0.1 mM X–0.0125 U/mL to 1 mM X–0.1 U/mL XO. Increase in spermatozoa motility parameters was recorded following treatment with antioxidants and seminal plasma. The level of the oxidative stress indices lipid peroxidation (LPO) and carbonyl derivatives of proteins (CP) was significantly reduced after addition of CAT, SOD, or GTH along with seminal plasma. Significant differences in SOD, glutathione reductase, and glutathione peroxidase activity were seen in spermatozoa incubated with, compared to that without, seminal plasma at all studied X–XO concentrations. The data demonstrated that CAT, SOD, or GTH in combination with SP can reduce reactive oxygen species stress in fish spermatozoa and improve spermatozoa quality.     

High zinc in the erythrocyte plasma membranes of common carp Cyprinus carpio

ABSTRACT:   The distribution of zinc (Zn) concentration in the blood and erythrocytes of common carp, grass carp, silver carp and tilapia was studied. It was found that in whole blood, the average Zn concentrations in these four species of fish (approx. 6–14 μg/[ml whole blood]) were relatively similar to those in other species of fish and mammals. However, the mean Zn concentration in the erythrocytes of common carp is approximately two times higher than the other three species of fish (approx. 5 vs approx. 2 μg/[ml whole blood]). It was found that approximately 70% of the Zn in the common carp whole blood came from its erythrocytes. In addition, approximately 43% of the Zn in the erythrocytes of common carp was found to be located on its outer plasma membranes. When an antibody against a 43 kDa Zn-binding protein, isolated from the digestive tract tissue of common carp, was used, significant quantities of the protein were shown to be present on the erythrocyte plasma membranes of common carp by an indirect immunofluorescent staining. High Zn on the outer plasma membrane of the common carp erythrocyte most probably comes from the 43 kDa Zn-binding protein.     

鲤与生长性状相关的EST-SSRs标记筛选

以大头鲤(Cyprinus pellegrini Tchang)(母本)与荷包红鲤(Cyprinus carpio wuyuanensis)抗寒品系(父本)杂交产生的F1作为亲本,以其自交F2作为分离群体,结合"拟测交"策略,用EST-SSRs标记对其中92个个体进行基因型检测.利用Windows Map Manager 2.0的标记回归法对符合拟测交分离的70个EST-SSRs标记进行单标记回归分析,检测出8个与鲤体长、体高、体厚性状相关的标记.对同一标记不同基因型个体间生长性状进行显著性比较,通过t检验,找到与生长性状相关的基因型.将上述8个EST序列与GenBank数据库进行BLAST比对,有3条EST序列与蛋白库中已知功能的序列高度同源.其中HLJE225与编码斑马鱼(J9enio rerio)冷诱导基因RNA结合蛋白的基因同源性水平高达97%,HLJE222与编码斑马鱼DAZ相关蛋白1基因同源性水平也高达97%,HLJE599与编码斑点叉尾鲴(Ictalurus punctatus)核糖体蛋白L6基因的同源水平为87%.本研究鉴定的与鲤生长性状相关的功能基因及有利用价值的基因型,为鲤重要生长性状的QTL(数量性状基闪座)定位和分子标记辅助育种提供了一定的依据.[中国水产科学,2009,16(1):15-22]     

Storage of common carp, Cyprinus carpio L., eggs for short durations

In this study, a short-term storage of pre-activated eggs of the Japanese ornamental (koi) carp, Cyprinus carpio L., at three different temperature regimes is reported. Koi eggs were stored at low temperatures (6-9^oC), at variable or high temperatures (12-31C) and at moderate-stable temperatures (20-24.5^oC). Survival of the developing embryos was examined at the first and the second day post-activation, and in hatch-out larvae. Survival was calculated for each treatment by linear regression Y = a-bX (P± 0.01), except for eggs incubated at temperature of about 20 C(P > 0.1).The specific mortality index (b/a 100), as a ratio between the mortality rate (b) and the fertilization rate (a), indicated that the highest mortality is associated with the high and unstable storage temperatures, while the lowest mortality is with the moderate and stable storage temperatures. Eggs can be stored at moderate and stable temperatures for a maximal duration of 6h, yielding survival of hatch-out larvae higher than 50%. In two trials, fertilization potency of sperm stored in a domestic refrigerator (5-9^oC) for 5h, was compared with freshly stripped sperm and no differences in fertilization rates were found between the two treatments.     

微卫星分子标记辅助镜鲤家系构建

用微卫星标记辅助进行镜鲤(Cyprinus carpio L.)家系的建立及选育,通过对亲本遗传结构及遗传差异的分析,预测产生优良家系的最佳配组。用28个微卫星分子标记评估松浦镜鲤亲本群体的遗传潜力,结果显示,群体的多态信息含量(PIC)达到0.5627,处于高度的遗传多样性水平,具备进一步繁育筛选优良群体的潜质。用x2检验和遗传偏离指数(d)估计Hardy-Weinberg平衡,结果表明群体处于平衡状态,但在16个位点表现出杂合子缺失,这可能跟长期的人工选择有关。依据个体间的遗传距离,连续2年共建立1对1亲本的繁育家系70个,用网箱分别养殖,2月龄家系体长均值为7.24～10.60cm,体质量均值为14.02～40.63g,方差分析显示家系间体长、体质量差异均极显著;对其中4个家系1龄鱼种生长情况及遗传结构分析也表明家系间的遗传分化较大,近交系数(Fst)达到0.1537,家系的多态信息含量在0.3056～0.4077之间,保持中度多态水平。由子代家系的生长和遗传结构两方面的实验结果证实了微卫星在辅助家系建立尤其是亲本选配方面具有较好的预测性,所获得的家系差异较大,具备进一步选育优良家系的潜力,从而证实了用此方法对现有镜鲤种质进行遗传改良是可行的。     

鲤胸腺素Tβ全长cDNA的克隆与序列分析

应用荧光DDRT-PCR从鲤外周血白细胞克隆了鲤胸腺素β(thymosinβ,Tβ)基因cDNA全序列。序列分析表明,TβcDNA全长528bp,其完整的读码框架位于40～178bp,编码46个氨基酸,5'非编码区长39bp,3'非编码区长为348bp,且具有Poly(A)加尾信号(AATAAA),将该基因序列递呈GenBank,注册序列号为AY457946。氨基酸序列同源性分析显示,鲤Tβ氨基酸序列与鲤Tβa、斑马鱼Tβa及斑马鱼Tβ-2同源性均为84%。在系统发生树上,鲤Tβ与鲤、斑马鱼、白斑狗鱼和鲑Tβa及斑马鱼Tβ2聚类。     

Effect of feeding Spirulina platensis on the growth, proximate composition and organoleptic quality of common carp, Cyprinus carpio L.

The effect of feeding Spirulina platensis on the growth, carcass composition, organoleptic quality, digestive enzyme activity and digestibility of common carp, Cyprinus carpio L., was studied through a culture trial lasting 120 days. Four experimental diets were employed by replacing fish meal protein from the standard diet at 25%, 50%, 75% and 100% through the incorporation of Spirulina. Another diet with Spirulina as the sole source of protein was also used. The final weight gain, specific growth rate, food conversion ratio and protein efficiency ratio of common carp were not affected by Spirulina supplementation. However, the diet with Spirulina as the sole source of protein resulted in better net protein retention. The muscle RNA:DNA ratio of fish fed Spirulina diets was higher than that of fish fed control diet. There was no significant difference in carcass moisture and protein content in the fish fed Spirulina diets as compared to fish-meal-based control diet. The carcass ash and fat contents were positively and negatively correlated with dietary Spirulina level, respectively. Organoleptic evaluation revealed no effect of Spirulina feeding on the quality of both raw and cooked fish. The gut digestive enzyme activity did not show any definite trend with respect to Spirulina supplementation. Spirulina improved the protein digestibility of the diets.     

鲤免疫应答相关基因的克隆与鉴定

为研究鲤(Cyprinus carpio L.)白细胞免疫应答相关的分子机理,以体外培养的鲤外周血白细胞为实验材料,用荧光标记的mRNA差异显示(FluoroDDRT-PCR)技术,研究丝裂原(50μg/mL LPS、50μg/mL PHA和50μg/mL ConA)在刺激白细胞4、12和24 h内诱导白细胞免疫应答相关基因的mRNA表达差异,共获得92个差异片段,其中87个片段有再扩增产物,再扩增率为94.6%;将差异片段克隆,经PCR鉴定,获得81个阳性克隆,鉴定率为93.1%;差异片段序列同源性功能分析结果表明,本研究共获得3个免疫应答相关的cDNA克隆,它们分别编码鲤的蛋白酶体激活因子PA28α亚基、翻译延伸因子(EF-1α)和基质金属蛋白酶13(Mmp13)部分序列,为进一步研究这些差异表达基因在鱼类免疫中的作用机制奠定了基础。     

Cortisol effects on the testicular androgen synthesizing capacity in common carp, Cyprinus carpio L.

Our previous studies on the effect of stress on pubertal development in carp have shown that repeated temperature changes caused an increase in cortisol levels and a retardation of the first waves of spermatogenesis. Identical effects, accompanied by a decrease in 11-ketotestosterone (11KT) plasma levels and the gonadosomatic index (GSI) were induced by cortisol administration via cortisol containing food pellets. The decrease in plasma 11KT is caused by a direct effect of cortisol on the steroid producing capacity of the testis, independent of luteinizing hormone (LH) levels. However, the mechanism through which cortisol interferes with testicular steroidogenesis is unknown. In the present study, we showed that in vitro physiological levels of cortisol can inhibit the conversion of 11β-hydroxyandrostenedione (OHA) into androstenetrione (OA), which is the precursor of 11KT, possibly by competing for the enzyme 11β-hydroxysteroid dehydrogenase (11β-HSD). The same mechanism may occur in vivo. However, our results demonstrate that an elevation of plasma cortisol levels during acute cortisol treatment did not result in lower plasma levels of OA and 11KT, but we did observe an accumulation of OHA. We suggest that the previously observed decrease in 11-oxygenated androgens, as an effect of long-term cortisol treatment, is caused by a retardation of testicular development. This results in a lower steroid synthesizing capacity of the testis as a whole. Although the in vitro observed cortisol inhibition of the conversion of OHA into 11KT plays a role in the accumulation of OHA, it apparently has no effect on the final 11KT plasma concentration. This revised version was published online in August 2006 with corrections to the Cover Date.     

Additive, dominance genetic effects for growth-related traits in common carp, Cyprinus carpio L.

Additive, dominance genetic effects were analysed for body weight (BW) and seven morphometric traits from three variants of red common carp, Cyprinus carpio var. singuonensis, C. carpio var. wuyuanensis and C. carpio var. color, based on a diallel cross‐mating design by using the additive–dominance genetic model. The results indicated that at 8 months of age (juvenile stage), the traits of pre‐dorsal height (PDH), pre‐dorsal width (PDW) and caudal peduncle height (CPH) were mainly controlled by additive effects; the traits of BW, total length (TL), standard length (SL), head length (HL) and caudal peduncle length (CPL) were mainly controlled by dominance effects. The narrow‐sense heritabilities of all traits ranged from 0.000 to 0.556; however, the broad‐sense heritabilities ranged from 0.453 to 0.775. At 20 months of age (adult stage), the traits of TL, SL, PDH and CPL were mainly controlled by additive effects, whereas the traits of BW, PDW, HL and CPH were mainly controlled by dominance effects. The narrow‐sense heritabilities of all traits ranged from 0.000 to 0.710; the broad‐sense heritabilities ranged from 0.629 to 0.934. Through the genetic merit prediction from three parents, it was found that the additive and dominance effects of C. carpio var. color could increase the BW and body length in their progeny, but these two genetic effects of C. carpio var. singuonensis and C. carpio var. wuyuanensis could decrease BW in their progeny. The genetic effects of C. carpio var. wuyuanensis could reduce total length, SL and caudal peduncle length, but increase body height in their progeny.     

Effects of microcystin-containing cyanobacterial extract on hematological and biochemical parameters of common carp (Cyprinus carpio L.)

The aim of the study was to assess the effects of a cyanobacterial extract containing microcystins (MCs) on selected hematological and biochemical parameters in common carp (Cyprinus carpio L.), as well as to determine the accumulation of toxins in fish tissues. The fish were immersed for 5 days in water containing toxins at a final concentration of 12 μg/L of microcystin LR equivalent. Microcystin LR residues were detected in fish liver, reaching 207, 238 and 260 ng/g f.w. of the tissues taken 24 h, 72 h and 5 days after the end of intoxication, respectively. The most substantial changes were found in fish plasma, including increases in creatine kinase, lactate dehydrogenase, ammonia, glucose, aspartate aminotransferase and alanine aminotransferase levels. A decline of about 50% in lysozyme activity was observed by the end of the experimental period. Moreover, a marked increase in ceruloplasmin activity was detected 24 h after the end of intoxication with a subsequent decrease in its activity after 72 h and 5 days. This study concludes that not only consumption of food containing toxins but also MCs dissolved in water may pose a threat to fish health. Additionally, detected changes in lysozyme and ceruloplasmin activity may have distinct effects in fish resistance against pathogens or oxidative stress, which should be taken into account in the future studies.     

Genetic characterization of wild Dutch common carp (Cyprinus carpio L.)

Six male carp, caught in the water system surrounding the Anna Paulowna (AP) Polder in The Netherlands, were characterized using allozyme and microsatellite markers. At the sMDH‐A1,2* loci an allele was found, which has previously only been found in wild River Rhine and wild Vietnamese common carp. Microsatellite allele frequencies showed that these AP carp were significantly different from a group of carp originating from several different domesticated strains. Based on both allozyme and microsatellite data, the AP carp probably originated from a wild or feral self‐sustaining population.     

Immunomodulatory effect of cimetidine in common carp (Cyprinus carpio L.)

The main indication of cimetidine is being H₂-receptor antagonist, but studies suggest that cimetidine may also act as a non-specific stimulant of cell-mediated immunity and immunomodulator. In order to determine the immunomodulatory effect of dietary intake of cimetidine in the common carp (100 ± 10 g), subjects were fed diets containing 0 (control), 50, 100 and 200 mg cimetidine kg^?1 of dry diet for a period of 6 weeks. TLC and NBT assays were significantly (P < 0.05) stimulated in cimetidine-supplemented groups displaying the highest value in 200 mg kg^?1 group. A decrease (P < 0.05) in cortisol and ACH50 value was recorded in fish treated with cimetidine. Serum protein, albumin and serum globulin levels were not significantly changed. The findings of the present investigation suggest that the incorporation of cimetidine in the diet of common carp enhances the non-specific immunity.