First report on the isolation of immunoglobulin M of guppy, Poecilia reticulata, for the production of polyclonal antibodies

As guppies are one of the commercially important fish in freshwater ornamental aquaculture industry, it is important to gain an understanding of guppy immune response for infectious disease control. Till now, the number of study that examined the immune response of guppies is limited and effective tools for monitoring guppy antibody have not been reported. In this study, we successfully isolated guppy IgM using mannan-binding protein (MBP) affinity chromatography and produced specific polyclonal antibodies against guppy IgM heavy and light chains, that showed a molecular weight of approximately 74 and 23 kDa respectively. The produced polyclonal antibodies were used to develop an enzyme-linked immunosorbent assay (ELISA) and have demonstrated to be an effective tool for the detection and quantification of antigen-specific antibody of guppies immunized with Pseudomonas fluorescens. In conclusion, the produced anti-guppy IgM polyclonal antibodies should prove its future implications for immunology and epidemiology studies in guppies.     

Classification of guppies’ (Poecilia reticulata) gender by computer vision

Labor costs of guppy growers and breeders are largely those of manual sorting (by strain, quality and gender) and counting fish. In most farms, female and male fish are grown together and sold either separately or together. Sorting fish according to gender is important for marketing as well as for breeding programs, so that a device for sorting and counting fish can potentially reduce production costs and improve quality.

A project aiming to develop sorting and counting technologies for ornamental fish growers included development and testing of image-processing algorithms for sorting guppy fish (Poecilia reticulata) by gender. The algorithms are derived from shape and color differences between female and male guppies. An algorithm for the determination of landmarks on fish contours was developed and found to be accurate in accordance with human judgment, enabling extraction of specific shape and color features of the tail and the body.

The algorithms were applied to three sets of images of guppies of the “Red-Blond” strain. Gender identification accuracy was approximately 90% using shape features, approximately 96% using color features and was slightly improved when both color and shape features were used.

Some of the components used are essential for future development of a computer vision based system for sorting and grading ornamental fish by strain and quality.     

Guidance of groups of guppies (Poecilia reticulata) to allow sorting by computer vision

Prior to marketing, ornamental tropical fish are currently sorted, graded and counted manually; a process that is labor intensive, stressful to fish and inaccurate. Manipulation of fish movements at our will is a key element in the development of a device for automatically sorting, grading and counting fish by a computer vision system, and the present study is one step towards the development of such a fish-friendly sorting device. The positive phototactic and rheotactic innate responses of guppies (Poecilia reticulata) were exploited to induce them to swim through transparent pipes or narrow channels, to render them ready for inspection by a computer vision system. Water flow direction and velocity, water level, and illumination were effective in inducing fish to move from one container into the other via a narrow transparent pipe. It was attempted to separate guppies moving in groups into individually spaced fish, by introducing an obstacle into a narrow channel or narrowing the transparent pipe.     

DNA markers for estimation of inbreeding depression and heterosis in the guppy Poecilia reticulata

Molecular markers have significant potential for use in precise breeding programmes in aquaculture. This paper reviews the use of DNA markers to estimate inbreeding depression and heterosis in the guppy Poecilia reticulata. Full‐sib matings revealed that inbreeding causes declines in survival and salinity tolerance, but not in undwarfism, growth and high water temperature tolerance, indicating the effects of inbreeding differ among fitness‐related traits. Salinity tolerance was used to quantify the level of inbreeding depression and heterosis because the trait is strongly sensitive to inbreeding and shows a linear decrease with an increase in inbreeding coefficient. A positive correlation was observed between heterozygosity at microsatellite loci and salinity tolerance among 17 guppy populations. This indicates that heterozygosity estimated from microsatellites is a useful indicator for the estimation of inbreeding depression, suggesting that overall heterozygosity is important for fitness‐related traits that show inbreeding depression. Use of DNA markers to estimate the amount of heterosis in various strain combinations was examined using diallele and reciprocal crosses among four domestic strains. The amount of heterosis differed among the strain combinations and correlated with Nei's genetic distance measured by microsatellites and also by dissimilarity using random amplified polymorphic DNA (RAPD) analysis. This indicates that microsatellite and RAPD markers are useful for estimating the amount of heterosis in various strain combinations, further suggesting that the amount of heterosis depends on the genetic differences between the strains. The present study showed that DNA markers are useful tools for estimating inbreeding depression and heterosis in guppy breeding.     

Culture of the guppy, Poecilia reticulata, in Singapore

Singapore is known internationally as a breeding centre for the guppy, Poecilia reticulata. About 30 domesticated colour pattern and tail shape varieties are reared, mainly for export. The culture of these varieties at two rural and eight resettled farms is described. All farms are small family concerns. The resettled farms practise monoculture of guppies while the rural ones integrated farming. Guppies are reared in large, shallow, outdoor, cement tanks and nylon cage-nets suspended in ponds. Water in aquaria and ponds has salinities from 0.5 to 9.0‰, at neutral or alkaline pH of 7.0 to 8.3. The sizes and stocking densities of aquaria and nets vary within and between farms. In breeding tanks, a male to female sex ratio of 1:3, 1:4 or 1:10 is maintained. Newborn fry are collected daily and placed in nursery aquaria. After 3 weeks, these fry are sexed: young males are transferred to grow-out cement tanks, and females to grow-out cagenets. Each farm has at least two ponds, one for water storage and the others for cagenet culture. Strains are cultured separately to maintain pure lines. Farmers improve colour patterns, size and fin shapes of the strains by continued mass selection. Four- to 6-month-old guppies are selected as brookstock and the rest which meet export quality are held in stocking aquaria and sold when demand arises.Each farmer prepares his own formulated diet for the fish. Supplemental foods like live tubificiids, commercial dried fish flakes, or floating pellets may be given to adults, and water fleas to fry. The rural farms add pig manure to fertilise the tanks.To remain viable in the future, farmers need to increase productivity by mechanising cleaning, draining, and filling tanks and ponds, feeding, and harvesting of the fish. This will cut down on labour and its increasing cost. Understanding the genetics of economically important characters will also help farmers produce higher quality and more exotic strains.     

栅栏技术在淡腌半干鲈鱼加工工艺中的应用

为确定在实验条件下大口黑鲈 (Micropterus salmoides) 幼鱼适合的麻醉方式与麻醉剂量,研究比较了7种不同质量浓度的MS-222 (40 mg·L^–1、50 mg·L^–1、60 mg·L^–1、70 mg·L^–1、80 mg·L^–1、90 mg·L^–1、100 mg·L^–1)和丁香酚(8 mg·L^–1、10 mg·L^–1、12 mg·L^–1、14 mg·L^–1、16 mg·L^–1、18 mg·L^–1、20 mg·L^–1) 2种常见渔用麻醉剂对体质量为(130±10) g的大口黑鲈幼鱼的静水麻醉效应。静水麻醉实验结果表明：1)随着2种麻醉剂浓度升高,幼鱼入麻时间缩短,复苏时间延长;2)在实验条件下,MS-222与丁香酚的最适麻醉质量浓度分别为70 mg·L^–1和16 mg·L^–1,最适运输质量浓度分别为50 mg·L^–1和10 mg·L^–1。使用最适运输浓度的MS-222和丁香酚对幼鱼进行10 h模拟运输,观察麻醉效应和分析血清生化指标发现：1)麻醉组运输成活率和复苏后24 h成活率均为100%,显著高于无麻醉运输组的80%和60% (P<0.05);2)较之无麻醉运输组和静水对照组,麻醉运输组的丙氨酸转氨酶(ALT)、天门冬氨酸转氨酶(AST)水平显著升高(P<0.05)。在麻醉运输组中,MS-222麻醉组的ALT和AST水平显著高于丁香酚麻醉组(P<0.05)。     

Estimation of the mode of inheritance of thermal tolerance in the guppy <Emphasis Type="Italic">Poecilia reticulata</Emphasis>

High water temperature influences the survival, growth, and maturation of fish. Genetically characterizing thermal tolerance is one of the most important subjects in fish culture. To identify the genetic characterization of thermal tolerance, this characteristic was compared among strains, and among parents and their offspring, in the guppy Poecilia reticulata. In the strain comparison, significant differences in survival rate were observed among the five strains examined, and between females and males. Females exhibited greater tolerance than males in four of five strains examined. In the comparison between parents and their offspring, stronger influence of female parent than of male parent was observed. Offspring obtained from surviving females exhibited greater tolerance than those from dead females. This tendency was typically observed in male offspring. The survival rate in male offspring obtained from dead female parents was lower than that of those from surviving females. The high-temperature tolerance of male parents did not influence this characteristic in offspring as strongly as that of female parents. These results suggest that the major gene or genes, which has a dominant resistant allele and a recessive sensitive allele, are probably passed on by sex-linked inheritance, located on the X chromosome.     

岱衢洋和官井洋大黄鱼自交与杂交子代生长性能及杂交优势分析

2007年3月至2008年8月对象山港海区网箱养殖的岱衢洋、官井洋大黄鱼自交以及正反向杂交子代进行生长特性观察。结果表明:整个实验过程(0~526 d),官井洋大黄鱼自交子代(GG)和反交子代(GD)(官井洋♀×岱衢洋♂)大黄鱼较岱衢洋自交子代(DD)和正交子代(DG)(岱衢洋♀×官井洋♂)大黄鱼生长快,到526 d实验结束时,GG和GD体重分别达到330.514 g和336.694 g,而DD和DG则仅为278.975 g和243.297 g。对其各阶段生长速度分析,发现虽然一龄DD生长较慢,但过冬后DD生长明显加快,体长增长速度超过其他各群子代,体重增长也达到GG和GD增长水平,即有后期增长潜能。对比各群子代肥满度发现,526 d时,DD肥满度最低(1.675),GD肥满度(1.779)虽高于DD,但显著小于GG(1.933)。鉴于GD生长速度高于DD,与GG相当,而体型又优于GG,认为GD具有较大的经济养殖和优良品种选育的潜力。拟合各阶段体长体重数据,得出各群体生长曲线公式如下,DD:W=0.0206L^2.9427, R²=0.9949;GG:W=0.0139L^3.0994,R²=0.9785;DG:W=0.0229L^2.9136,R²=0.9905;GD:W=0.0177L^3.0079,R²=0.9949,拟合度较好,可通过体长估算体重,应用于实际生产。     

“吉富”系列罗非鱼生长性状的配合力、杂种优势与遗传相关分析

"吉富"系列罗非鱼是中国当前养殖的主要罗非鱼种类。该研究以3个品系"吉富"尼罗罗非鱼(Oreochromis niloticus)(新吉富、吉富、吉诺玛)为研究材料,应用3×3完全双列杂交设计构建了9个组合,采用数量遗传学方法对各组合子一代的体质量、全长、体长、体高和体宽5个生长性状的配合力、杂种优势与遗传相关进行了分析。结果表明:5个生长性状均未检测到一般配合力,而特殊配合力效应达62%以上,表明这些性状主要受到特殊配合力的影响;进一步的杂种优势分析发现吉诺玛♀×新吉富♂、新吉富♀×吉诺玛♂、吉富♀×新吉富♂这3个组合的杂种效果明显,其中以吉诺玛♀×新吉富♂为最佳组合;5个生长性状之间均存在显著的遗传相关与表型相关(P0.05),可进行多性状的聚合利用。该研究结果为"吉富"系列罗非鱼的配套利用提供了有益的资料和数据,同时为开展罗非鱼配套系育种提供了依据。     

Inbreeding evaluation using microsatellite and its effect on growth traits in turbot,Scophthalmus maximus

Turbot is an economically relevant fish, and simultaneously at the risk of inbreeding depression in China. Microsatellite DNA markers were used to evaluate inbreeding level of a random turbot breeding population and further investigate its effects on growth traits. DNA based inbreeding coefficients (F_DNA) were ranged from 0 to 0.4124. Average inbreeding coefficient was 0.0621 with a standard deviation of 0.0658. Regression analysis results showed inbreeding had a very significant (p < .01) effect on harvest body weight (BW) and significant (p < .05) effect on harvest body long (BL). Inbreeding depression, expressed as the percent change in phenotype per 10% increase in F, was ?4.00% for BW and ?1.14% for BL. The inbreeding depression on high inbreeding coefficient (more than 10%) individuals was further analyzed. A set of significant (p < .05) and larger inbreeding depression was obtained, ?7.15% and ?2.49% for BW and BL respectively. This study demonstrates that inbreeding has a negative effect on growth traits and highlights the need of DNA markers to control inbreeding and maximize the genetic diversity in selection breeding programs.     

中华卤虫不同日龄生长性状间的相关性分析

为探究中华卤虫不同生长阶段以及性状间的相关性,以确定合适的选择时间和目标性状,2008年收集来源于中国不同地区的3个两性生殖品系卤虫卵,SIN品系(山西运城解池盐湖)、YIM品系(内蒙古伊克昭盟)和ALXZ品系(内蒙古阿拉善左旗),利用人工静水孵化技术进行孵化,每个品系随机选取50尾个体单独养殖30d,每5天测定卤虫个体的全长(TL),并在第25日龄、第30日龄测定全长(TL)、体长(BL)、体宽(BDW)和体重(BW)性状值。中华卤虫3个品系6个生长阶段的各性状值测定数据显示,25d是卤虫生长的最高峰。不同生长阶段间全长表型相关分析表明,两个生长阶段间隔时间越长,相关系数越小。SIN、YIM和ALXZ3个品系第5天和第25天的相关系数仅为0.073、0.091、0.026。特别是15d前和15d后的生长阶段间,生长性状值甚至存在着负相关。对于卤虫生长性状的遗传改良,早期选择特别是15d之前进行选择,可能并不是一个理想的选择方案。中华卤虫性状间相关分析表明,第25日龄、30日龄全长、体长分别与体宽、体重性状间的表型相关虽均达到极显著水平(P<0.01),但相关系数较小。在选育生长快、体型大的卤虫新品种(系)时...     

瓯江彩鲤配套选育系繁殖性状的配合力测定与杂交优势分析

以瓯江彩鲤(Cyprinus carpio var.color)5个配套选育系进行双列杂交产生的F1的数据为分析对象,利用统计学和数量遗传学方法对6个繁殖性状(雄鱼性腺指数、雌鱼性腺指数、绝对繁殖力、相对繁殖力、受精率和孵化率)进行了配合力测定和杂交优势分析。结果发现,不同组合间的繁殖性状存在较大差异,但以"麻花"作母本的交配组合的平均雌鱼性腺指数和孵化率最高,以"全红"作母本的交配组合的绝对繁殖力和相对繁殖力最高,以"粉花"作母本的交配组合的所有繁殖性状参数均较差;雌鱼性腺指数与绝对繁殖力、相对繁殖力之间均存在显著的表型相关(r=0.411～0.847,P=0.01～0.04),绝对繁殖力与相对繁殖力之间存在极显著的表型相关(r=0.847,P<0.01);雌鱼性腺指数、绝对繁殖力和相对繁殖力3个性状检测到显著的特殊配合力;通过遗传效应预测,10个杂交组合均存在特殊配合力与杂交优势预测值,其雄鱼性腺指数、雌鱼性腺指数、绝对繁殖力、相对繁殖力和孵化率的杂交优势与特殊配合力均呈极显著相关(r=0.959～0.985,P<0.01)。本研究结果为瓯江彩鲤配套系育种提供了参考数据。     

不同贝龄栉孔扇贝数量性状的相关性和通径分析

为深入了解栉孔扇贝形态性状与湿重之间的关系,为栉孔扇贝选育工作中亲贝的挑选提供策略,本研究使用来自同一家系的栉孔扇贝子代324只一龄个体以及生长到二龄的230只个体的性状数据进行通径分析。性状数据包括壳长(x_1/cm),壳高(x_2/cm),壳宽(x_3/cm),湿重(y/g)。结果显示,所有形态性状和湿重之间的相关系数均达到极显著水平。其中与一龄贝湿重相关系数最大的是壳长,为0.939,与二龄贝湿重相关系数最大的是壳高,为0.808。通径分析结果显示,壳长对一龄贝湿重的直接影响最大(0.532),壳高对一龄贝湿重的直接影响最小(0.163)。壳高对二龄贝湿重的直接影响最大(0.451),壳长对二龄贝湿重的直接影响最小(0.191),决定系数与以上通径分析结果的变化趋势一致。利用多元回归的方法构建了一龄贝和二龄贝形态性状与湿重间的回归方程,一龄贝:y=–10.527+0.287x_1+0.087x_2+0.409x_3,R~2=0.926;二龄贝:y=–68.609+0.254x_1+0.719x_2+2.008x_3,R~2=0.830。本研究结果为栉孔扇贝种贝的挑选提供了理论依据。     

草鱼幼鱼生长性状和肌肉成分的遗传参数估计

为了研究草鱼幼鱼阶段生长性状和肌肉成分的遗传改良潜力。随机采集288尾人工繁育的4月龄草鱼幼鱼,基于12对微卫星标记,鉴定出来自16个家系（8个母本,9个父本）的273尾个体,各家系、母本和父本对应子代贡献率存在极显著差异（P<0.01）。对草鱼3个生长性状（体质量、体长和肥满度）和2个肌肉成分指标（粗蛋白含量和粗脂肪含量）比较发现,3个生长性状和粗蛋白含量在家系和母系半同胞间存在显著差异（P<0.05）,而3个生长性状和2个肌肉成分指标在父系半同胞间差异均不显著（P>0.05）。基于动物模型和限制性最大似然法,草鱼4月龄体质量、体长、肥满度、粗蛋白含量和粗脂肪含量的遗传力估值分别为0.34、0.33、0.17、0.17和0.20,其中3个生长性状遗传力估值统计检验显著（P<0.05）。遗传相关分析显示,体质量与体长之间呈极显著的高度正相关（0.82, P<0.01）,肥满度与体质量和2个肌肉成分之间存在显著的正相关（0.17-0.29, P<0.05）。研究表明,草鱼4月龄生长性状具有较高的选育潜力,并可通过体长选择实现对体质量的遗传改良;推测在草鱼生长改良过程中,可能会伴随肥满度和粗脂肪含量的变化,这应当引起重视。     

条斑紫菜双单倍体群体主要经济性状的遗传分析

条斑紫菜叶状体主要经济性状的遗传参数及相互间的遗传关系,是开展分子育种的基础。实验以条斑紫菜野生型品系(Py-WT2,父本)和红色突变型品系(Py-HT,母本)杂交后产生的杂合丝状体为材料,构建由152个品系组成的条斑紫菜双单倍体(DH)群体。该群体叶状体的6个经济性状(L50、W50、FW50、SGR-L、SGR-W、SGR-FW)的表型值用单样本Kolmogorov-Smirnov检验。结果发现,各性状均为数量性状。L50和SGR-W为超亲遗传性状,其余4个性状的变异介于双亲之间,其中W50偏向于母本,FW50、SGRL和SGR-FW偏向于父本。6个性状的变异系数介于21.11%~56.68%,均属中等强度变异。相关性分析表明,L50、W50和FW50相互之间均存在极显著正相关性,SGR-L、SGRW和SGR-FW相互之间也存在极显著正相关性。利用单因素方差分析法估算出L50、W50和FW50的遗传力分别为58.17%、64.00%和57.64%,控制3个性状的基因对数分别为6.61、12.63和8.09,遗传力(y)与基因对数(x)的二次回归方程为y=0.2922x~2–4.6533x+76.162(R~2=1)。基因间互作方式的检测结果显示,控制L50和控制FW50的多基因间均分别不存在互作;控制W50和控制SGR-W的多基因间均分别存在互补作用;控制SGR-L和控制SGR-FW的多基因间均分别存在重叠作用。研究表明,条斑紫菜叶状体L50和FW50的遗传力高、基因对数少且基因间无互作,可进行早代选择。另外,L50、W50和FW50之间的相关性高,可进行间接选择以提高育种效率。     

星康吉鳗形态性状与体重的通径分析及生长曲线拟合

为探明星康吉鳗（Conger myriaster）形态性状和体重的关系,测定了体重（Y）及全长（X₁）、肛长（X₂）、尾长（X₃）、头长（X₄）、吻长（X₅）、眼径（X₆）、眼后头长（X₇）、背鳍前长（X₈）、躯干长（X₉）、头宽（X₁₀）、眼间距（X₁₁）等11个形态性状,通过相关分析、通径分析、回归分析等方法研究了形态性状对体重的影响;通过曲线拟合获得4个形态性状与体重的最佳拟合模型。结果表明,星康吉鳗各形态性状之间以及形态性状与体重之间均呈极显著正相关关系（P< 0.01）。相关分析发现肛长（X₂）与体重的相关系数最大（0.970）;通径分析发现肛长（X₂）对体重的直接作用最大（0.450）,头宽（X₁₀）通过肛长（X₂）对体重的间接作用最大（0.431）;决定系数分析发现,肛长（X₂）对体重的直接决定系数最大（0.203）,肛长（X₂）和头宽（X₁₀）的共同决定系数最大（0.172）。形态性状（X）与体重（Y）的多元回归方程为Y=-113.859+6.481X₂+49.213X₆+11.514X₇+20.145X₁₀（R²=0.954）。星康吉鳗肛长（X₂）、眼后头长（X₇）、头宽（X₁₀）与体重的最佳拟合模型为指数函数,模型方程分别为Y=1.466e^0.287X、Y=1.435e^1.328X、Y=1.970e^1.974X,眼径（X₆）与体重的最佳拟合模型为线性,方程为Y=-77.460+222.093X。结论认为,在星康吉鳗选育时,应以肛长（X₂）和头宽（X₁₀）为主要选择性状,以眼径（X₆）和眼后头长（X₇）为辅助选择性状。     

草鱼生长相关的微卫星标记在选育群体中的验证

生长性状是水产动物遗传育种中的重要经济性状,利用与性状相关的分子标记与育种相结合的手段,可以大大加速育种进程。在对草鱼生长性状的前期研究中,采用数量性状位点(QTL)定位的方法,在1号连锁群中发现了2个与生长相关的QTL。在此基础上,实验利用这2个QTL侧翼的2对微卫星标记(CID391₂、CID1512、CID973₁和CID254₁),对长江草鱼选育群体的480个个体进行分析,以期基于草鱼QTL定位结果,对草鱼生长相关的微卫星标记在选育群体中进行验证。结果显示:(1)4个微卫星标记在该群体中均具有高度多态性,其中各位点观测等位基因数(N_a)为12～23个,有效等位基因数(N_e)为4～12个,观测杂合度(H_o)为0.607～0.904,期望杂合度(H_e)为0.751～0.902;(2)利用方差分析及多重比较对4个多态性的微卫星标记与选育草鱼群体的生长性状(体质量和体长)进行关联分析,发现CID391₂在雌性个体...     

长丰鲢生长性状相关微卫星标记的初步筛选

为了筛选出与长丰鲢生长性状相关的分子标记,以指导其种质鉴定和选择育种工作,实验利用15个多态微卫星标记,对6、17和36月龄长丰鲢进行了遗传多样性和生长性状关联分析.结果 显示,各月龄长丰鲢的观测杂合度(Ho)和期望杂合度(Hc)均高于0.5,PIC均值分别为0.555、0.445和0.490,表明各月龄长丰鲢群体均具...     

褐点石斑鱼不同月龄形态性状的主成分及通径分析

为研究褐点石斑鱼不同月龄形态性状的生长规律特征及形态性状对体质量的影响,采用主成分分析和通径分析方法对3月龄、8月龄和13月龄褐点石斑鱼个体全长(X_1)、体长(X_2)、头长(X_3)、体宽(X_4)、体高(X_5)、尾柄高(X_6)、眼径(X_7)、眼后头长(X_8)和体质量(Y)9个性状进行分析。结果显示,(1)不同月龄褐点石斑鱼各形态性状对体质量的相关系数均达到极显著水平,不同月龄相同形态性状与体质量的相关性存在差异。(2)各月龄第一主成分均指向褐点石斑鱼的生长发育情况;第二主成分均指向眼睛发育情况;但3月龄、8月龄和13月龄第三主成分分别指向尾部发育、体宽发育及体高发育。(3)3月龄阶段,全长、体高、体宽及尾柄高对体质量的直接通径系数达到显著水平;8月龄阶段,全长、体宽、尾柄高及眼后头长对体质量的直接通径系数达到显著水平;13月龄阶段,全长、体高、体宽及体长对体质量的直接通径系数达到极显著水平,并建立了3个月龄组的不同形态性状对体质量最优回归方程。(4)褐点石斑鱼3~13月龄阶段的体长(L)与体质量(W)的关系为W=0.0378L2.873(R~2=0.9596),说明该阶段生长属于等速生长。研究表明,在褐点石斑鱼的不同生长阶段,各形态性状对体质量的影响有所不同,建议以全长(3~13月龄)作为育种工作的目标性状,为褐点石斑鱼早期生长发育阶段的选择育种工作和测量指标提供科学依据。