Effect of feeding apidaecin on common carp (Cyprinus carpio) growth performances and immune function

Effect of apidaecin on common carp (Cyprinus carpio) based on growth performance, feed utilization (Feed Conversion Ratio, FCR) and immune parameters were investigated. Twelve aquaria with three replicates for G1-3 and control were used. The synthesized apidaecin peptide was added to common carp basal diets (Control) as additives in three concentrations: 10.0 mg kg^− 1 (G1), 15.0 mg kg^− 1 (G2) and 30.0 mg kg^− 1 (G3) by wet weight of basal diet. After an 80-day feeding experiment, G2 and G3 showed significantly better (P < 0.05) results of growth performances and FCR than the control. However, there was no remarkable difference (P > 0.05) between G2 and G3. As for G1, the final weight, SGR (Specific Growth Rate) and DG (Daily Gain) were increased and the FCR was decreased, but not significantly (P > 0.05) compared with the control. The serum lysozyme activity of G2 and G3 was significantly increased (P < 0.05) than the control. As for the alternative complement pathway activity, G2 and G3 were also higher (P < 0.05) than the control. However, no significant difference (P > 0.05) was found whichever between G2 and G3 or between G1 and G2 in both lysozyme and the alternative complement pathway activities. Furthermore, there was no remarkable difference between G1 and the control in immune parameters.     

氧化应激对鲤抗氧化状态和免疫功能的影响

为探讨氧化应激对鲤抗氧化状态和免疫功能的影响,本实验以H_2O_2作为活性氧自由基(ROS),将鲤暴露于不同浓度的H_2O_2(0、0.25、0.50和1.00 mmol/L)中,诱导鲤产生氧化应激反应。连续暴露7 d后,采集鲤血液和肝组织,以检测相关生化指标以及基因表达量的变化。结果显示,与空白对照组(0 mmol/L)相比,随着H_2O_2浓度的升高,血清葡萄糖(GLU)、皮质醇(cortisol)和乳酸(LA)含量显著升高;而碱性磷酸酶(AKP)和酸性磷酸酶(ACP)活性仅在1.00 mmol/L H_2O_2处理组中显著高于其他实验组。氧化应激参数显示,与空白对照组(0 mmol/L)相比,0.50和1.00 mmol/L H2O2处理显著降低血清过氧化氢酶(CAT)活性,而提高还原型谷胱甘肽(GSH)、丙二醛(MDA)和总抗氧化能力(T-AOC)水平;在肝脏组织中,1.00 mmol/L H_2O_2处理显著降低了GSH含量,促进了MDA生成。基因表达结果显示,与空白对照组相比,1.00 mmol/L H2O2处理组显著上调了肝脏组织中cyp1a表达,而下调了cyp1b表达;同时0.50和1.00 mmol/L H2O2处理显著上调了hsp70、hsp90、c3、c-lyz和hep的表达。研究表明,氧化应激暴露可诱导鲤产生明显应激反应和脂质过氧化,降低机体抗氧化能力并激发免疫应答反应。     

Increased stocking density influences the acute physiological stress response of common carp Cyprinus carpio (L.)

The physiological response of common carp, Cyprinus carpio (L.) to increased stocking density and an additional acute net confinement stressor was investigated. Stocking densities were increased from 28.4 to 56.8 or 113.6 kg m^?3 by the use of crowding screens and fish were sampled from the crowded groups after 15, 39 and 87 hours of crowding (hc). A transient elevation of plasma cortisol was found in the higher density group after 15 hc before values returned to control levels. Increased stocking density also increased plasma levels of glucose, free fatty acids (FFA) and lactate during the experimental period. No effect was found on oxygen radical production in the blood, haemoglobin or haematocrit levels, but leucocrit values were lower after 39 hc in both groups. Net confinement resulted in a significant increase in plasma cortisol levels in all groups. However, after 15 hc, cortisol and FFA levels in both crowded plus confinement groups were higher than in the control and confined groups. Confinement resulted in increased oxygen radical production in the crowded plus confined groups at all times. Results indicate that although carp responded with a mild stress response to increased stocking density and adaptation occurred by 87 hc, the crowded fish were more sensitive to an additional acute stressor.     

福瑞鲤2号不同生长速率个体肌肉组织转录组分析

鱼类通过选育获得优良生长性状的遗传分子机制 目前尚不清楚.实验分析比较了 2种不同生长速率福瑞鲤2号肌肉组织的转录组文库.组装后共获得3 92 23 8条测序序列(contigs).其中可比对上斑马鱼、弓斑东方鲀、青鳉、三刺鱼、尼罗罗非鱼和松浦镜鲤的蛋白序列的比例为56.01％～77.71％.2种不同生长速率福瑞鲤2号...     

鲤胰蛋白酶原基因的电子克隆及生物信息学分析

为深入研究胰蛋白酶在鱼类中的生理功能和作用机制,利用生物信息学的方法,成功获得了鲤3种胰凝乳蛋白酶原cDNA序列(ccTRP1、ccTRP2和ccTRP3)并对其进行了序列分析。结果表明,三者均含有一个长度为729bp的开放阅读框(open reading frame,ORF),编码由242个氨基酸组成的胰蛋白酶原,其中包括15个氨基酸组成的信号肽和5个氨基酸(LDDDK)组成的激活肽。总平均亲水性GRAVY(grand average of hydropathicity)分析表明,三者均是亲水性蛋白。氨基酸比对结果显示,三者具备胰蛋白酶原的保守结构特征,如含有催化三联体氨基酸(His-57、Asp-102和Ser-195),12个半胱氨酸,位于底物结合口袋底部Asp-189和口袋开口处的Gly-216、Gly-226等。同时,鲤3种胰蛋白酶之间具有90%以上的同源性。进化树结果显示,鲤3种胰蛋白酶均属于Group I(阴离子型胰蛋白酶),且三者的进化距离不一致。     

Effects of dietary medium-chain triacylglycerols (tricaprylin and tricaproin) and phospholipid supply on survival, growth and lipid metabolism in common carp (Cyprinus carpio L.) larvae

The present study investigated the interaction of dietary medium-chain fatty acids (MCFA) and phospholipids (PL) on survival, growth and lipid metabolism in common carp larvae. Nine diets based on casein and dextrin and with a variable lipid part were tested in triplicate for 22 days post first feeding. The 3×3 design consisted of three triacylglycerols (3% of diet) combined with three different lipid supplements. Tested triacylglycerols were triolein (TOL), tricaprylin (TC8) and tricaproin (TC6), and lipid supplements were 2% soybean oil (low-fat diets without PL), 2% soybean lecithin (low-fat diets with 2% PL) or both 2% soybean lecithin and 6% TOL (high-fat diets with 2% PL).

In the first step, both TC6 and TC8 resulted in improved survival and growth rates compared to TOL, irrespective of the PL supply. In the second step, TC8 decreased survival and growth rates, whereas the difference between TC6 and TOL became less. Histological signs of impaired intestinal absorption of neutral lipids were evidenced in larvae fed TOL without PL and also in high-fat diets with 2% PL. The latter diets also resulted in poorer growth rates compared to low-fat diets with 2% PL. These results suggest that the quantitative PL requirement of larvae increases as the dietary level of long-chain triacylglycerols increases. Larvae fed TC6 or TC8 showed enlarged liver and hepatocyte volume and a decreased level of body neutral lipids. Based on β-hydroxybutyrate (β-HBA) measurements in whole larvae, TC8 was found to be more ketogenic than TC6. TC6 and TC8 affected differently the fatty acid profile of larval body neutral lipids. TC6 did not induce the appearance of MCFA, whereas TC8 feeding resulted in a low level of 8:0 and relatively high levels of 10:0 (3.8% of total fatty acids). Neither 8:0 nor 10:0 were found in larval polar lipids.

This study confirmed the essentiality of PL in common carp larval diets and underlines differences in the utilization of TC6 and TC8, which both initially stimulate growth during the first week, but only temporarily in the case of TC8.     

利用EST-SSR分子标记研究鲤的饲料转化率性状

饲料转化率是重要的经济性状,通过QTL定位获得的紧密连锁标记以及相关基因,是遗传育种的重要工具。本文利用70个鲤的EST-SSR,对鲤全同胞家系92个体的基因组DNA进行扫描,得到符合1∶1孟德尔分离类型的位点48个,3∶1孟德尔分离类型的位点22个。采用拟回交策略对符合1∶1分离的标记与鲤饲料转化率性状进行单标记回归分析。结果表明：HLJE335、HLJE253、HLE547、HLJE92、HLJE203、HLJE231等6个标记与饲料转化率性状相关（P<0.05）。其中,HLJE547和HLJE203两个标记与饲料转化率相关性达极显著水平（P<0.001）,分别解释了表型变异的11.00%、11.00%。应用NCBI数据库,分别对与性状极显著相关的2个ESTSSR标记进行同源性比对,发现标记HLJE547与斑马鱼基因组中编码ATPase, H⁺ transporting, lysosomal的核酸序列同源（同源性为74%）,与斑马鱼编码的ATPase, H⁺ transporting, lysosomal的蛋白序列同源（同源性为92%）。推测ATPase可能通过影响鲤消化与吸收等生物学机制,从而影响鲤的饲料转化效率。     

Heritability estimates for processing and quality traits in common carp (Cyprinus carpio L.) using a molecular pedigree

Heritability estimates with a microsatellite parentage assignment based pedigree are of special interest in common carp cultured under traditional pond conditions. This method reduces common environment effects as all families can be grown immediately after hatching mixed in the same pond. We applied this method to study genetics of growth and processing traits in common carp at market size (1.5 kg and more). The experimental progeny was established by crossing 147 two-year old males and 8 females (six to eight-year old) of Hungarian synthetic mirror carp population. The fish grew up through three vegetation seasons and at the end a sample of 331 fish was examined for biometrical traits (standard length, body weight, relative head length, relative body height, relative body width), percent fat and processing traits (% processed body, % fillets with skin, % fillets without skin). It was shown that sex had a significant effect on most traits: females were larger and fatter than males, and they had both higher percent processed body and percent fillet with skin. Standard length, body weight, percent fat and relative head length had a high heritability (> 0.5), while relative body height, relative body width, percent processed body and fillet yields had a medium heritability (0.2–0.5). We found relatively high positive genetic correlations between body size (standard length and body weight) and percent fat (0.71 and 0.59, respectively), favourable genetic correlations between body size and percent processed body (0.69 for standard length and 0.74 for body weight) and between body size and fillet yields (0.50–0.77). Genetic correlations between body size and body shape (relative head length, relative body height and relative body width) were weak to moderate, thus selection for better growth should have little impact towards more rotund shape. Relative head length had strong negative correlation (− 0.7 to − 0.9) with percent fat, percent processed body and percent fillet yields. This means that indirect selection for reduced relative head length should be effective in improving of fillet yield.     

池塘和稻田养殖模式对金边鲤和建鲤肌肉品质的影响

分析了稻田和池塘养殖模式下金边鲤和建鲤肌肉营养成分、理化特性和质构特性,并进行营养价值评价,评估金边鲤的肌肉品质,为开发和推广金边鲤稻田养殖提供数据依据。结果显示,稻田金边鲤的粗脂肪、粗灰分和系水力均显著高于池塘金边鲤;稻田建鲤和池塘建鲤的基本营养成分除水分外差异均不显著;稻田金边鲤肌肉的硬度、弹性、咀嚼性显著高于池塘建鲤和稻田建鲤,硬度和恢复性显著高于池塘金边鲤,而内聚性显著低于池塘金边鲤、池塘建鲤和稻田建鲤;测定的17种氨基酸中,4组鱼肉的Lys含量均高于WHO/FAO标准和鸡蛋蛋白标准,稻田金边鲤的∑TAA、∑EAA、∑DAA和∑NEAA均高于池塘建鲤和稻田建鲤,且EAA/TAA比值更接近FAO/WHO要求的40%的标准;稻田和池塘金边鲤的EAAI均超过100分,高于稻田和池塘建鲤(82.31～83.36分);测定出的24种脂肪酸中,稻田金边鲤的∑SFA、∑MUFA和DHA+LA+油酸总量均高于其他3组。研究表明,金边鲤和建鲤鱼肉可作为人体优质的赖氨酸源,稻田金边鲤肌肉比建鲤和池塘金边鲤肌肉更富有嚼劲,其肌肉必需氨基酸、不饱和脂肪酸及高度不饱和脂肪酸含量更高,且组成比例更符合人体需求。稻田养殖金边鲤的肌肉品质优势明显,具有较高的产业开发潜力。     

辛基酚对鲤的雌激素效应

研究了辛基酚对鲤的雌激素效应。经10、50、100、300和500μg/L辛基酚暴露32d后,鲤存活率和肥满度与对照组无差异;性腺指数变化明显,雌鱼性腺指数随暴露剂量的增大而增大,雄鱼性腺指数随暴露剂量的增大而减小;50μg/L及以上暴露组与对照组差异显著(P<0.05)。辛基酚能诱导鲤雄鱼产生卵黄蛋白原,暴露剂量为10μg/L组有部分雄鱼肝脏匀浆和血液中检出卵黄蛋白原,50、100和300μg/L组卵黄蛋白原含量随辛基酚浓度的增大而显著增大,500μg/L组卵黄蛋白原含量低于300μg/L组、高于100μg/L组,均为极显著差异(P<0.01)。     

建鲤与异育银鲫生长、生化组成和饲料利用的比较

In order to investigate the difference in utilization of diets with different quality in Jian carp（Cyprinus carpio var. Jian）and allogynogenetic silver curcian carp(Carassius auratus gibelio), a 55 d growth trial was conducted and low quality diet (LQ-diet) and high quality diet (HQ diet) were tested. LQ-diet contained 33.91% dietary protein which is mainly from soybean meal while HQ-diet contained 45.59% dietary protein which is mainly from fish meal. The initial average body weights were from 5.58 g to 5.82 g for two fish strains. The trial was carried out in a system consisting of 12 self circulation 320 L tanks. During the experiment, the fish were fed to satiation twice a day (at 9: 00 and 15:00), and uneaten feed was collected 1 h after feeding and dried. Feces were collected twice a day (at 11:00 and 16:45) from the fecal traps and dried at 70 ℃. The results show that feed intake was higher in Jian carp than in allogynogenetic silver curcian carp when fed LQ diet, while there was no significant difference between weight gain, feed conversion efficiency, protein efficiency rate and apparent digestibility. When fed HQ diet, Jian carp showed a lower feed intake, but higher feed conversion efficiency and protein efficiency rate than allogynogenetic silver curcian carp while there was no significant difference in the weight gain and apparent digestibility of both species. For Jian carp, feed intake and protein efficiency rate for the fish fed HQ diet and LQ diet were not significantly different, while the fish fed HQ diet showed higher weight gain, feed conve rsion efficiency and apparent digestibility. For allogynogenetic silver curcian carp, the fish fed HQ diet showed significantly higher feed intake, apparent digestibility, weight gain, feed conversion efficiency and lower protein efficiency rate. For Jian carp, body contents of dry matter, protein, lipid and energy for the fish fed HQ diet and LQ diet were significantly higher. For allogynogenetic silver curcian carp, body contents of dry matter and protein was significantly higher, while body contents of lipid and energy were affected by diet qualities. Compared to Jian carp, allogynogenetic silver curcian carp showed better utilization when fed LQ diet while poorer utilization when fed HQ diet.     

Effect of water temperature and short-term fasting on macronutrient self-selection by common carp (Cyprinus carpio)

The influence of water temperature (17 and 25 °C) and a period of fasting on the self-selection of macronutrients were examined in common carp held on a 12:12-h light/dark cycle. Eight replicate groups of juvenile carp (ca. 15 g body weight at 17 °C and 25 g at 25 °C) were fed with three test diets [high-protein (HP), high-fat (HF) and high-carbohydrate (HC) diets] by individual self-feeders for 3 weeks, followed by a 2-week period of fasting and then 3 weeks of refeeding. At 17 °C, the carp initially selected the HP and HF diets [HP/HF/HC (%)=38:40:22], whereas at 25 °C, the carp selected the HP diet over the others (55:21:24). After fasting, the carp tended to increase their demand for the HF diet at 17 °C and for the HC diet at 25 °C, but the overall self-selection of the test diets did not differ markedly from that prior to fasting. Carp held at 17 °C had higher relative whole body fat content and plasma triglyceride concentration; the latter decreased markedly during fasting. These results suggest that water temperature influences the macronutrient selection by carp: a relatively high demand for both protein and fat at 17 °C and for protein at 25 °C. Two weeks of fasting, however, did not markedly alter their macronutrient selection.     

The in vitro effect of CpG-ODNs on the innate immune response of common carp, Cyprinus carpio L.

Unmethylated CpG dinucleotides within bacterial DNA or synthetic oligodeoxynucleotides (ODNs) can activate immune cells. A panel of synthetic oligodeoxynucleotides was evaluated in vitro for their ability to enhance the innate immune response of common carp (Cyprinus carpio L.). In vitro addition of CpG-ODNs enhanced the phagocytic activity and superoxide anion production of stimulated kidney phagocytes. The CpG-ODNs also induced lymphocyte proliferation in the fish kidney cells. The ODNs containing multiple CpGs generally resulted in greater stimulatory capacity, although CpGs located at the terminus of an ODN were ineffective. These results indicated that CpG-ODNs enhance the innate immune response of common carp.     

中国鲤育种的主要方法、遗传解析及展望

水产种业是水产养殖业发展的基础,是渔业战略性、基础性核心产业,也是保障未来养殖业绿色、健康发展的核心竞争力。随着水产业全球化、市场化的发展,我国种业正面临着前所未有的机遇与挑战。特别是随着生物技术的快速发展,水产育种也由传统的选择育种和杂交育种,发展至细胞工程育种、分子标记辅助育种、全基因组选择育种、分子设计育种和基因组编辑等精准设计育种。水产动物重要经济性状的基础研究及其遗传改良技术的创建驱动着我国水产种业的蓬勃发展,截止2022年,通过全国水产原种和良种审定委员会审定并由农业农村部公告的水产新品种就有266个,其中鲤新品种数量最多,为31个,表明鲤育种工作卓有成效。本文重点回顾了我国鲤的种质和基因组资源现状,鲤的主要品种及其育种方法;简要介绍了鲤生长、抗病、体色、饲料转化率等经济性状关联的遗传研究进展,并由此提出了新时代背景下鲤种业的发展方向和措施建议,以期为我国鱼类育种提供借鉴。     

中国鲤育种的主要方法、遗传解析及展望

为探究鲤不同体色个体肌肉品质的差异,本研究以福瑞鲤2号青灰色个体 (BG)和全红个体 (WR)为对象,通过检测其肌肉色泽、氨基酸、脂肪酸和质构指标,评价这2种体色福瑞鲤2号肌肉品质的差异。结果显示,BG的b^*值显著高于WR;出肉率以及肌肉的水分、粗脂肪、粗蛋白和灰分无显著差异;不同体色福瑞鲤2号的肌肉中均能检测出17种氨基酸,其中人体必需氨基酸7种,非必需氨基酸10种。甘氨酸 (Gly)、脯氨酸 (Pro)、组氨酸 (His)和半胱氨酸 (Cys)在2组间差异不显著,其他氨基酸在2组间存在显著或极显著差异,两种体色福瑞鲤2号之间ΣEAA/ΣTAA没有显著差异;BG肉豆蔻酸 (C14:0)显著高于WR,BG的饱和脂肪酸含量较WR稍高,WR的单不饱和脂肪酸含量较BG稍高,二者多不饱和脂肪酸含量相近;WR的弹性和咀嚼性显著高于BG。研究表明,体色对福瑞鲤2号的肌肉品质有影响。本研究为鱼类肉质性状改良提供基础数据和参照依据。

     

嗜水气单胞菌刺激下鲤血清质子转运载体15a2(Slc15a2)的表达分析

为解析鲤质子转运载体15a2 (Slc15a2)在嗜水气单胞菌侵染过程中对机体免疫应答机制的影响,实验通过制备鼠源Slc15a2-1和Slc15a2-2多克隆抗体,采用酶联免疫吸附测定(ELISA)技术检测嗜水气单胞菌感染后鲤血清中Slc15a2-1和Slc15a2-2蛋白表达变化。结果显示,(1)感染后2个拷贝的最高蛋白表达水平与空白对照组相比,分别增加了4.81和2.48倍;与Slc15a2-1相比Slc15a2-2的表达量从0～48 h一直处于较高表达水平。(2)从整体来看,Slc15a2的2个拷贝具有相似的表达趋势,均为先升高后降低、再升高再降低;但Slc15a2-1在3 h达到第1个表达高峰,而Slc15a2-2则在6 h达到第1个表达高峰。(3) 6～24 h的4个时期中,Slc15a2-1和Slc15a2-2蛋白表达呈现互补的表达趋势,即在同一时期一个拷贝的表达量显著下降,另一个拷贝的表达量显著升高,但总体维持较高表达水平。结果表明,本实验制备的鼠源Slc15a2-1和Slc15a2-2多克隆抗体具有较高的亲和力和特异性,当鲤受到嗜水气单胞菌侵染后,2个拷贝整体具有相似的...     

Efficiency of local feeds for the common carp (Cyprinus carpio L.) in Saudi Arabia

The effects of feeding three local feeds (53%, 43% and 33% protein) on the growth and quality of the common carp (Cyprinus carpio L.) were investigated. The 43% protein feed proved most economical, with the highest biological value and has increased the protein and amino acid contents of the flesh but resulted in poor fish growth (0.10 g/day).     

茜素络合物对鲤仔鱼耳石标记特征研究

利用100 mg/L的茜素络合物（alizarin complexone,ALC）对鲤仔鱼进行48 h的水环境浸泡标记,以探讨该ALC标记方法的特征,及其对矢耳石、星耳石和微耳石的标记效果以及鱼体ALC浸泡、续养恢复与耳石ALC标记区域形成和消失的时滞进行研究。结果显示,3种耳石在可见光和荧光下均能检测到明显的标记环。其中星耳石的标记效果最佳,微耳石次之。耳石上荧光信号出现和消失的时间与鱼体ALC浸泡开始和结束的时间均存在1 d的时滞。此外,浸泡标记过的鲤仔鱼在进行了长达50 d的续养恢复后,其耳石上的ALC标记环仍清晰可见。研究表明,ALC标记法所形成的标记环在耳石上可长期存在,使用ALC对鲤仔鱼进行生态标记具有很强的可行性。     

清水江鲤基于微卫星标记和形态指标的遗传分析

利用12个微卫星标记对鲤(Cyprinus carpio)的4个野生群体 [清水江鲤、太湖鲤、黄河鲤(C.carpio haematopterus)和黑龙江鲤(C.carpio amurensis)] 和2个选育群体 [福瑞鲤(C.carpio var. FFRC)和松浦镜鲤(C. carpio var. specularis 'Song-pu')] 共208尾个体进行遗传分析。结果显示,12个位点共检测到341个等位基因,平均等位基因数为28.67,其中1个位点(HLJ1127)检测到正向选择压力;选育群体的遗传多样性参数普遍低于野生群体,其中松浦镜鲤群体的各项参数均值最低(N_a=6.82,H_o=0.54,PIC=0.50),清水江鲤群体的各项参数均值最高(N_a=21.25,H_o=0.80,PIC=0.91);分子方差分析显示,整体遗传变异主要来自群体内,但群体间呈极显著遗传分化(P<0.01);基于群体Nei's遗传距离的UPGMA聚类树和PCoA分析表明,鲤4个野生群体间遗传距离较近,而与2个人工选育群体间遗传距离较远;基于个体遗传结构及PCoA分析显示部分野生个体遗传结构比较混杂,而选育个体的遗传结构则相对单一。研究表明,中国鲤野生资源具有较高的遗传多态性,而人工选育群体维持着较纯的遗传种质。