Isolation and characterization of bacteria associated with a syndrome disease of sea urchin Strongylocentrotus intermedius in North China

Sea urchin, Strongylocentrotus intermedius, transplanted from Japan in 1989, has been widely cultured along the coasts of Liaoning and Shandong Provinces and has become the dominant and most economically important maricultured species in North China. However, a lesion syndrome symptom of S. intermedius broke out frequently these years, showing lethargy in activities, blackish peristomial membrane and body well lesions, and brought about high mortality eventually. Six representative prominent bacterial strains were isolated from diseased sea urchin from September 2009 to January 2010. By means of API 20NE and 16S rRNA gene sequences analysis, isolates were identified as Shewanella aquimarina, Pseudoalteromonas tetraodonis, Vibrio shilonii, V. harveyi, V. fortis and V. splendidus. Bacterial challenge tests showed that their representative isolates were virulent to S. intermedius with LD₅₀ values ranging from 9.2 × 10⁴ to 3.4 × 10⁶ CFU/g body weight, among which S. aquimarina , V. fortis and P. tetraodonis were highly virulent, and the other three isolates showed moderate virulence. The results indicated that a variety of bacteria including Shewanella, Pseudoalteromonas and Vibrio were involved in the mortality of S. intermedius, and the six isolates were opportunistic pathogens of sea urchins. All isolates reported herein were sensitive to ampicillin, enrofloxacin, ofloxacin, doxycycline and florfenicol.     

Estimates of heritabilities and genetic correlations for growth and gonad traits in the sea urchin Strongylocentrotus intermedius

The present study is the first estimate for the sea urchin, Strongylocentrotus intermedius, of genetic parameters for growth and gonad traits during the entire reproductive period. 2165–5764 sea urchins were sampled from 74 to 120 families at six different growth stages. Six hundred individuals were sampled from 20 randomly chosen families at harvest. Coefficients of variation for most growth and gonad traits were high (18.08–78.66%) suggesting a good variation foundation for breeding. Point estimate for heritabilities based on the sire components of variance were moderate to high for test height (0.24–0.39), test diameter (0.21–0.48) and body weight (0.16–0.49). Heritabilities of gonad wet weight, gonad index and gonad moisture at harvest were 0.17, 0.41 and 0.50 respectively (moderate or high). Gonad colour parameters, L^* (lightness) and a^* (redness) have a heritability of 0, while b^* (yellowness) and colour difference ΔE₁ and ΔE₂ have very similar heritabilities of 0.2, 0.18 and 0.21 respectively. Heritabilities of β‐carotene concentration and content were 0.89 and 0.44 respectively. The genetic correlations between the two measured traits ranged from ?0.984 to 0.999. Gonad wet weight, L^*, b^*, β‐carotene concentration and β‐carotene content had significant positive genetic correlations with all the growth traits. However, gonad index, gonad moisture and colour differences ΔE₁ and ΔE₂ had significant negative genetic correlations with growth traits. The present study indicates that growth, gonad yield and most gonad quality traits could be improved by selection with moderate to high potential. Indirect selection should be applied on gonad traits using growth traits as secondary traits. The present study provides important theoretical guidance for future selective breeding programmes for S. intermedius.     

中间球海胆幼体及稚海胆生长性状的遗传参数估计

为估计中间球海胆Strongylocentrotus intermedius幼体及稚海胆生长性状的遗传参数,采用巢式不平衡设计方法建立了44个中间球海胆全同胞家系,并测量了各家系海胆四腕幼体、六腕幼体和八腕幼体等各期浮游幼体的体长以及受精后30 d的稚海胆壳径,采用动物模型和约束性最大似然法(REML)估计了中间球海胆幼体和稚海胆生长性状的遗传参数。结果表明:中间球海胆四腕幼体和八腕幼体体长的遗传力分别为0.705±0.373和0.538±0.444,为高度遗传力,六腕幼体体长和稚海胆壳径的遗传力均接近于0,为低度遗传力;各阶段幼体体长和稚海胆壳径的共同环境效应为0.048~0.352;各阶段幼体体长间遗传相关为正相关(0.300~0.518),各阶段幼体体长与稚海胆壳径间的遗传相关为负相关(-0.152~-0.435)。本研究结果可为制定中间球海胆的早期选择和间接选择选育方案提供参考依据。     

虾夷马粪海胆群体的遗传多样性及微卫星标记与生长性状的相关性分析

为了明确性腺性状和分子标记之间的关系,在海胆选育工作中提高性腺性状,利用微卫星标记对海胆各生长性状及性腺颜色进行相关分析。选用28个虾夷马粪海胆多态性微卫星分子标记对虾夷马粪海胆自交F1代家系群体的进行了检测分析,结果表明:共检测到80个等位基因,平均等位基因数为2.857 1个;平均有效等位基因数为0.827 0个; 观测杂合度平均值0.515 0,期望杂合度平均值为0.530 4;平均多态信息含量（PIC）为0.402 6,卡方检验显示其中有9个位点发生了偏离(P<0.01)。采用一般线性模型(GLM)进行连锁显著性检验,发现与壳高、体重和体积显著相关（P<0.05）的微卫星标记各1个,分别为TS131、TS38和TS105;与海胆颜色L*值（暗-亮）显著和极显著相关的微卫星标记分别为4个 (TS38、TS85、TS105、TS108)和1个(TS41);与海胆颜色a*值（绿-红）显著和极显著相关的微卫星标记各1个(TS101 TS41);与海胆颜色b*值（蓝-黄）显著相关的微卫星标记3个(TS41、TS114和TS128);与海胆口器重显著和极显著相关的微卫星标记各1个(TS114和TS46)。该研究结果可以为虾夷马粪海胆的相关性状进一步QTL定位提供参考数据,并为虾夷马粪海胆的养殖和选育提供理论指导。     

湛江近海海域裸体方格星虫消化系统的剖析

本文为探索湛江近海海域裸体方格星虫的消化系统特点,以湛江近海海域裸体方格星虫为材料,对其消化道的形态和组织学进行了切片剖析。结果表明,裸体方格星虫消化系统由口、咽、食道、肠、直肠、盲肠和肛门等组成;在直肠上附有直肠盲囊,其消化道约为体长的2.5倍,肠道通过3个回折及螺旋缠绕形成肠索;消化道管壁由内向外分为黏膜层、黏膜下层和浆膜层,黏膜层有大量的皱褶,没有连续肌肉层,而黏膜下层由结缔组织组成,含有肌纤维和血管神经等;裸体方格星虫消化道有发达的纤毛,直肠盲囊连接于直肠末端,里面充满黄色液体。     

饥饿和再投喂对中间球海胆性腺发育的影响

对中间球海胆Strongylocentrotus intermedius进行了不同时间的饥饿和再投喂试验。采用各组“饥饿时间 投喂时间”相同的试验设计,在30 d内,试验组(S3、S6、S9、S12组)海胆分别饥饿3、6、9和12d后再连续饱食投喂直至试验结束,同时设对照组S0。结果表明:试验结束时S3组海胆性腺重、性腺指数均接近对照组,其余各组较对照组低;特殊生长率在再投喂的第2 d或第4 d出现最大值。性腺组织学观察的结果表明:对照组海胆性腺为滤泡型,由生殖细胞和营养细胞构成,发育至生长期;随着饥饿时间的延长,雄海胆性腺中成熟精子的数量减少,雌海胆性腺中初级卵母细胞发育迟缓,而雌、雄海胆性腺中营养细胞的数量无明显变化;再投喂后,性腺有不同程度的发育。     

皱纹盘鲍与光棘球海胆筏式混养研究

皱纹盘鲍与光棘球海胆的混养可有效地防治污损生物的附着。综合分析鲍与海胆的生长、存活及防污损生物附着的效果,在生长期、高温期和恢复生长期混养海胆的最适密度分别为１５．２０个／格和１５－２０个／格。     

皱纹盘鲍与光棘球海胆越冬期筏式混养的初步研究

实验分越冬前期和越冬后期两个阶段进行。越冬前期鲍与海胆在生长方面的结果无显著差异,但混养可防止苔藓虫的附着,海胆的混养效果以２０个／格略好;越冬后期海胆混养的密度则以５个－１０个／格为最佳,混养可有效地防止多种敌害生物附着。     

三种海胆性腺总脂的脂肪酸组成的研究

本文采用了ＦＦＡＰ石英毛细和气相色谱柱,对光棘球海胆,虾夷马粪海胆及海刺猬的性腺中总脂的脂肪酸组成进行了研究。结果表明这几种海胆含有四十种以上的脂肪酸,分布十分相似。主要的ＨＵＦＡ为１８：４ｎ－３（３．２％－７．４）,２０：４ｎ－６（６．４％－１６．７％）及２０：５ｎ－３（６．６％－１２．１％）,并含有非常罕见的ｎ－５及ｎ－７烯酸。     

光棘球海胆(♀)×中间球海胆(♂)杂交育苗试验

以光棘球海胆为母本,中间球海胆为父本进行杂交试验及苗种培育.试验结果表明,通过筛选同时得到自然成熟的父母本精、卵,在比正常海胆自交精子量高40倍以上的精子作用下,杂交受精率为2.5%.在适宜的水温、光照及饵料等的培养条件下,杂交海胆能够生长附着变态,变态时间与母本相近.培育出的杂交海胆外部颜色及棘色、棘长介于两亲本之间.杂交海胆经过6～7个月的中间暂养,出苗量为0.6×10~4个/m~3,个体壳径3～30 mm,平均壳径8.8 mm.