长牡蛎"海大3号"生长繁殖与营养成分的周年变化

为阐明长牡蛎"海大3号"新品种的生长与繁殖特性,自2016年11月份至2017年10月份对养殖于山东省荣成海区长牡蛎"海大3号"群体的生长、性腺发育及营养成分的周年变化及其与环境因子的关系进行研究。结果显示,在海区水温较低的冬季1—2月份,长牡蛎"海大3号"生长缓慢,性腺处于休止期;随水温升高,在春季3—4月份配子开始发育,雌雄个体发育基本同步,4—5月份壳高生长速率加快,湿重显著增加;在夏季,随配子发育壳高生长速率降低,7月份因配子排放湿重下降;在产卵后的秋季长牡蛎"海大3号"壳高和湿重均显著增长。营养分析结果表明,随性腺发育,外套膜、性腺-内脏团、鳃和闭壳肌中糖原含量呈下降趋势,为配子发育提供能量;脂质和蛋白质作为配子主要结构物质在性腺中不断累积,随配子排放而显著降低;在秋季,4个组织中较高的蛋白质含量为机体的快速生长提供物质基础。同时外套膜作为构成壳物质的分泌器官,在8—9月份其糖原含量和脂质含量显著增加,这暗示机体的快速生长需要一定能量储备。研究表明,长牡蛎"海大3号"的生长和繁殖活动受海区水温季节性变化的影响,并与各组织营养成分密切相关。春季和秋季为机体快速生长时期,夏季为配子集中排放期;糖原是生长繁殖活动的主要能量来源,脂质是配子主要结构物质,蛋白质是机体产后恢复和快速生长的物质基础。     

长牡蛎壳金选育群体性腺发育与营养成分的周年变化

为了评估长牡蛎(Crassostrea gigas)壳金选育群体的繁殖周期及营养价值,自2013年11月至2014年10月,对山东省刘公岛海区长牡蛎第三代壳金选育群体的性腺发育、营养成分(总蛋白、总脂肪、糖原)季节变化及其与环境因子的关系进行了研究,并比较了与普通养殖群体营养成分的差异。结果表明,长牡蛎壳金选育群体雌雄比例接近1︰1,性腺同步发育;配子发生开始于2月,全年只有1个产卵季节(6―8月),7月为两性配子排放高峰期。配子成熟及排放期间,水温处于较高水平,食物充足,可为性腺发育提供能量,也有利于幼体的存活和生长。卵母细胞直径从配子发生开始逐渐增大,并在产卵前达到最大值(36.88μm),产卵后则降低。营养成分分析表明,脂肪含量在性腺–内脏团中随配子发育积累储存,产卵后显著降低;糖原在性腺发育到4月时开始下降,为配子的发育提供能量;蛋白质在配子发育过程中大量合成,与繁殖活动存在密切联系。长牡蛎壳金选育群体外套膜的总蛋白含量显著高于普通养殖群体(P0.05),糖原和总脂肪含量显著低于普通养殖群体(P0.05),其他组织的一般营养成分未出现显著差异。研究表明,长牡蛎在壳金群体选育过程中营养成分已表现出分化,这为长牡蛎壳色选育提供了重要参考依据。     

不同育性三倍体长牡蛎性腺发育过程中的营养成分比较

为阐明不同育性三倍体长牡蛎性腺发育与营养成分变化的关系,实验对不育型和可育型三倍体长牡蛎性腺发育过程中的主要营养成分(糖原、总蛋白质和总脂肪含量)进行分析,并与二倍体长牡蛎进行比较。结果显示,三倍体长牡蛎性腺—内脏团、闭壳肌和外套膜3种组织中的糖原含量均显著高于同时期的二倍体长牡蛎,性腺—内脏团和闭壳肌中的总蛋白质含量则显著低于同时期的二倍体。随着性腺发育,可育型三倍体长牡蛎性腺—内脏团的糖原含量下降了31.88%,二倍体长牡蛎下降82.41%,而不可育型三倍体长牡蛎糖原含量下降了0.55%,这与糖原为配子发生供能密切相关。此外,不育型三倍体长牡蛎性腺—内脏团的糖原、总蛋白质和总脂肪含量在繁殖季节中均没有发现明显的波动,而可育型三倍体长牡蛎由于性腺一定程度的发育,其营养成分含量的变化趋势与二倍体类似。研究表明不育型和可育型三倍体长牡蛎在繁殖季节营养成分存在明显的差异,不育型三倍体的糖原品质性状优于可育型,这为长牡蛎育性控制育种提供了重要参考依据。     

魁蚶生殖生物学初步研究

通过组织学观察,结合人工育苗实践,魁蚶的生殖周期可分为5期,即形成期、生长期(小生长期和大生长期)、成熟与排放期、退化期和休止期。魁蚶性腺在春天水温回升至4℃以上时开时发育;性腺发育的最适温度为14～20℃;产卵季节是6月下旬～8月下旬,7月为产卵盛期;产卵适温20～25℃。     

山东沿海脉红螺的性腺发育和生殖周期

为阐明脉红螺的繁殖生物学特性,于2012年3月至2013年2月对中国山东荣成靖海湾的脉红螺(Rapana venosa)的生殖周期及其与环境因子的关系进行了研究。组织学观察表明,脉红螺全年只有1个生殖周期,且雌雄配子同步发育,其性腺发育过程可分为5个时期:形成期,增殖期,成熟期,排放期,恢复期。配子发生始于9月,大部分个体在5月至6月成熟,排放的高峰期为7月。条件指数和平均卵径随配子发生而增大,成熟期达到最高值,产卵后减少。雌雄比例整体符合自然界性比为1∶1的自然规律。环境因子的分析结果表明,水温和叶绿素a含量存在明显的季节变化,而盐度无明显季节变化。相关性分析表明,水温与叶绿素a含量显著正相关(P0.05)。研究结果丰富了脉红螺的繁殖生物学资料,对脉红螺野生资源的管理保护、开发利用提供了重要的科学依据。     

魁蚶常温育苗技术

魁蚶（Scapharcabroughtonii），俗称赤贝，是一种大型、经济价值较高的贝类，其软体部的加工品“赤贝肉”，具有较高的出口创汇价值。近几年，我们利用夏秋季育苗场相对空闲时间进行魁蚶常温育苗，取得了较好效果。一、亲贝选择１．亲贝的采捕时间因不同海区水温和饵料条件的差异，魁蚶的性腺发育程度也不尽相同，一般在海区底层水温升至２０～２５℃时性腺逐渐进入成熟排放期。我国的黄海北部海区７～８月份为产卵盛期，故常温育苗多在７～８月份采捕亲贝。２．亲贝的选择方法选择壳长７～１０厘米、外形完整无损伤的魁蚶作亲贝。解剖检…     

魁蚶海区采苗技术

魁蚶俗称赤贝,属软体动物门、瓣鳃纲、蚶目、蚶科、毛蚶属,是一种营埋栖生活的大型双壳贝类,具有较高的经济价值。魁蚶的海区采苗技术对降低室内育苗生产成本,促进魁蚶养殖业的发展有着积极的意义。在黄海北部海区,大连市金州区登沙河镇养殖场的内湾,人们发现扇贝笼内有魁蚶苗种,便开展了魁蚶采苗试验。笔者经过近两年采苗工作实践,认为魁蚶海区采苗的主要技术如下:一、生物学调查1.性腺指数的测定魁蚶属雌雄异体,性腺在表皮与内脏囊之间,在繁殖季节雌性卵巢呈桔红色,雄性精巢呈桔黄色。成熟的卵为圆形,卵径约60μm。从6月中旬开始测定魁蚶…     

5种壳色长牡蛎不同组织常规营养成分分析与评价

对壳黑、壳紫、壳橙、壳金和壳白5种壳色长牡蛎(Crassostrea gigas)不同组织包括闭壳肌、外套膜、鳃、性腺-内脏团的营养成分(水分、总蛋白质、总脂肪、糖原、灰分)进行检测分析,比较它们与普通养殖群体营养成分的差异以及5种壳色长牡蛎各组织间营养成分的差异。结果表明,在闭壳肌中,壳紫长牡蛎的总脂肪含量显著高于壳橙长牡蛎(P0.05),其他组分在6个群体中未表现出显著性差异(P0.05);在外套膜中,壳金长牡蛎总脂肪含量显著高于壳黑、壳橙和普通养殖群体(P0.05),壳白长牡蛎的灰分含量显著高于普通养殖群体(P0.05),其他组分在各群体之间未表现出显著性差异(P0.05);在鳃中,长牡蛎壳橙、壳紫、壳白选育群体的糖原含量显著低于对照组(P0.05),其他组分在6个群体之间没有显著性差异(P0.05)。长牡蛎5种壳色选育群体和普通养殖群体的性腺-内脏团在水分、总蛋白质、总脂肪、糖原和灰分中均无显著性差异(P0.05)。4个组织中水分和灰分均以外套膜和鳃中含量最高,闭壳肌中总蛋白质的含量则在4个组织中居首位,总脂肪和糖原含量均以性腺-内脏团中最高。研究结果表明不同壳色长牡蛎之间以及不同壳色与普通对照群体之间在一些重要营养成分上已表现出分化,这为长牡蛎壳色品系选育提供了重要的参考资料。     

山东沿海中国蛤蜊的繁殖生物学特性

本研究于自2008年4月至2009年3月对中国山东海阳沿海中国蛤蜊(Mactra chinensis)的繁殖周期及其与环境因子(温度、盐度和叶绿素a含量)的关系进行了研究。山东沿海的中国蛤蜊全年只有1个繁殖周期,可分为6个时期:休止期,形成期、增殖期、成熟期、排放期、耗尽期。组织学观察显示,配子发育周期分为2个明显的阶段:11月至翌年2月的休止期和包括配子形成、成熟和排放的性腺发育期。该地区中国蛤蜊雌雄配子的发育时间基本同步,配子发生均始于3月,大部分个体在6月至7月成熟,8月雌雄配子大规模排放。中国蛤蜊的卵母细胞直径呈显著的周年变化(P0.05),从3月开始增加,7月达最大值。随着产卵的发生,卵径逐渐减小,至11月降至最低。条件指数在配子发生期开始增大,成熟期达到最高值,产卵后减小。     

虾夷扇贝组织中微量元素的分布特性

研究了虾夷扇贝闭壳肌、外套膜、内脏团、瓣鳃和性腺中Cu、Fe、Mn、Zn、Pb和Cd等元素的分布特性。试验结果表明,内脏团中的Cu显著(P<0.05)高于其他各组织,其他各组织间差异不显著(P>0.05);闭壳肌中Zn的含量显著(P<0.05)高于内脏团、外套膜、瓣鳃和性腺中的含量;Fe含量内脏团和瓣鳃中显著(P<0.05)高于性腺、闭壳肌和外套膜;各组织中Mn的分布特性为瓣鳃>闭壳肌>性腺>内脏团>外套膜;Pb的分布特性为内脏团>闭壳肌>性腺>外套膜>瓣鳃;内脏团中的Cd占全贝总量的67%,显著高于其他组织。因此,约占全贝质量10%的内脏团蓄积了较高含量的Cu、Fe、Pb和Cd,尤其是Cu和Cd(分别约占全贝蓄积总量的71%和67%),食用时去掉内脏团,可保证虾夷扇贝的食用安全和较高的食用及营养价值。     

Susceptibility by amberjack (Seriola dumerili), yellowtail (S. quinqueradiata) and Japanese flounder (Paralichthys olivaceus) to Neobenedenia girellae (Monogenea) infection and their acquired protection

This study investigated: 1) susceptibility differences to infection by Neobenedenia girellae (Capsalidae) between amberjack Seriola dumerili (Carangidae), yellowtail S. quinqueradiata and Japanese flounder Paralichthys olivaceus (Paralichthyidae); 2) growth and egg production of N. girellae on each fish species; 3) acquired protection of each fish species against this parasite. The number of N. girellae on S. dumerili was significantly higher than on S. quinqueradiata and P. olivaceus when these fishes were exposed to oncomiracidia in the same aquarium. Neobenedenia girellae growth on S. dumerili was fastest and, thus the number of eggs laid by parasites on S. dumerili was greater than on the other two species. Seriola dumerili and P. olivaceus, which were previously infected with N. girellae and treated by freshwater bath, acquired partial protection against re-infection by N. girellae. The relative re-infection of three S. dumerili individuals out of eleven individuals was markedly low compared with the initial infection, and the relative initial infection and re-infection on two P. olivaceus out of eleven individuals was markedly low. The results of this study could be useful to control N. girellae infections when cultivating S. dumerili, S. quinqueradiata and P. olivaceus.     

AFLP-based genetic linkage map of marine shrimp Penaeus (Fenneropenaeus) chinensis

This paper presents the genetic linkage map of the Chinese shrimp Penaeus (Fenneropenaeus) chinensis constructed with 472 AFLP markers. A hundred F₁ progeny from an intercross between a female from the new variety “Yellow Sea No. 1” and wild caught male used for the mapping study. Two separate maps were constructed for each parent. The female linkage map consisted of 197 marker loci forming 35 linkage groups and spanned a total length of 2191.1 cM, with an average marker space of 13.5 cM. The male map consisted of 194 marker loci mapped to 36 linkage groups and spanned a total length of 1737.3 cM, with an average marker spacing of 11.0 cM. The level of segregation distortion observed in this study was 12.2%. The estimated genome length of P. chinensis was 3150.3 cM for the female and 2549.3 cM for the male, respectively. The observed genome coverage was 69.6% for the female and 68.1% for the male map. The linkage maps constructed in this study provide basic information for further linkage studies on Chinese shrimp, and more importantly, the construction of the maps are part of the work of the genetic breeding programs which will be used for growth discovered in the QTL analysis of P. chinensis.     

太平洋鳕RAG1和IgM基因荧光定量PCR方法的建立及初步应用

为研究太平洋鳕发育早期特异免疫系统形成的机制,通过RAG1和IgM基因的转录水平衡量特异免疫系统的发育特点.根据GenBank中RAG1和IgM的序列信息,分别设计1对特异引物,从太平洋鳕头肾中扩增得到RAG1和IgM的基因片段.将所获基因片段分别插入到克隆载体pMD18-T中,从而构建太平洋鳕RAG1和IgM基因的质粒标准品.建立并优化太平洋鳕RAG1和IgM基因绝对荧光定量PCR方法.为进一步验证该方法的可靠性,分别利用绝对定量和相对定量检验目的基因在太平洋鳕早期发育过程不同组织内的表达差异.以优化后的绝对荧光定量PCR方法检测不同发育时期太平洋鳕RAG1和IgM的表达情况.结果显示,RAG1的回归方程为y=-3.266x+33.77,回归系数R2=0.996;IgM的回归方程为y=-3.119x +27.61,回归系数R2 =0.998.绝对定量和相对定量结果在基因转录趋势上显现出一致性,即RAG1基因在胸腺和头肾中表达,且在胸腺中的表达量显著高于头肾中的表达量,在肝脏和脾脏中无表达;IgM基因在胸腺、头肾、肝脏和脾脏中均有表达,其中脾脏中表达量最高,其次是头肾.RAG1基因在太平洋鳕发育早期的表达水平很低,到61日龄(days posthatching,dph)至95 dph表达量显著提高;IgM基因在早期表达水平同样很低,到33 dph至61 dph才有明显表达,在95 dph时表达量显著提高.研究表明,本实验方法可靠,特异性较强,可成功对目标基因转录水平进行检测.     

Recruitment and effects of Discocotyle sagittata (Monogenea) infection on farmed trout

Farmed rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta were monitored over 3 years for infection with the blood-feeding gill fluke Discocotyle sagittata. Parasite transmission is seasonal: new infections take place during summer/autumn, and transmission is generally negligible during winter/spring. There are 2 sources of infection for naïve fish-of-the-year: limited invasion when fish are in the raceways by riverborne larvae originating external to the farm; and internally, within the farm, when 0+ fish are transferred to ponds previously occupied by older cohorts of infected fish. Thereafter, infection levels continue to increase in rainbow trout primarily through transmission within the farm. Prevalence rose to 100% in 1+ fish by the end of their second summer. In O. mykiss, mean abundance reached 194 worms/host for 1+ fish (up to 489 worms/host) and 160 worms/host for 2+ fish. By contrast, in S. trutta, parasite prevalence never exceeded 85% and, after the first year's invasions, infection levels decreased over time: in 1+ and 2+ brown trout, parasite mean abundance was < 4 (maximum 15) worms/host. We present evidence of the detrimental effects of D. sagittata on the host: high burdens are associated with pale gills, decreased body condition and host mortality. Parasite burdens become overdispersed during the warmer part of the year, as prevalence and mean abundance increase. However, the degree of parasite overdispersion decreases over winter; we cannot distinguish whether decreased aggregation is due to parasite losses from infected fish (including immune-mediated parasite mortality) or parasite-induced host mortality.     

吉富罗非鱼与奥利亚罗非鱼自繁与杂交 F1遗传特性与抗病力分析

以吉富罗非鱼与奥利亚罗非鱼为繁育亲本,采用完全双列杂交繁育4组F1,将初始规格基本一致的4组罗非鱼饲养100 d后,运用“加性-显性”遗传分析模型,分析了4组F1罗非鱼8个生长相关性状杂种优势、遗传效应以及与性状间的相关性.结果表明:(1)F1群体平均优势为0.129 4～0.368 4.除尾柄长超亲优势较大外,其他性状的群体超亲优势较小或表现出负向超亲优势.(2)8个性状的广义遗传率(HB)为0.714 2～0.995 3,表明加性效应和显性效应对性状的遗传变异影响极显著(P＜0.01).除尾柄长外,其他性状的狭义遗传率(HN)介于0.469 4～0.737 9,表明加性遗传方差在表型方差中所占比率较高.(3)体质量、全长、体长、体高、体宽、头长、尾柄长、尾柄高性状之间表型相关在0.776 6～0.999 7范围内,而遗传相关在0.994 1～1.000 0之间,表明这些性状间都存在极显著的正相关.取样结束后,采用3.95×106 CFU/mL的海豚链球菌菌液进行腹腔感染,吉富罗非鱼自繁组F1代12 h后出现死亡,而奥利亚罗非鱼自繁组F1192 h后才出现死亡.384 h后,吉富罗非鱼自繁组F1死亡率为40％,正反交组F1分别为20％和23.3％,奥利亚罗非鱼自繁组F1死亡率最低,为6.67％.研究结果表明,除尾柄长外,杂交F1的其他性状不具备超亲优势,然而杂交可以提高选育后代的抗病力.     

4种岩礁性鱼类视网膜感光细胞和最小分辨角的比较

运用组织学方法,对铠平鲉、花斑平鲉、大泷六线鱼和斑头六线鱼的性成熟个体视网膜光感受细胞和最小分辨角进行了比较研究。结果表明:(1)大泷六线鱼和斑头六线鱼视锥细胞的排列方式为4个双锥细胞围绕一个单锥细胞构成正方形或者菱形镶嵌结构,铠平鲉和花斑平鲉的双锥细胞排列成行,单锥细胞随机分布于4个双锥细胞之间;分析认为,大泷六线鱼和斑头六线鱼视锥排列方式更复杂,单锥细胞与双锥细胞的比值(SC/DC)更高,推测其具有更好的色觉。(2)铠平鲉和花斑平鲉视柱细胞与视锥细胞的比值(R/C)大于7∶1,而大泷六线鱼和斑头六线鱼的R/C小于3∶1;推测铠平鲉和花斑平鲉更适合在夜间活动,而大泷六线鱼和斑头六线鱼属白天活动的鱼类。(3)铠平鲉、花斑平鲉、大泷六线鱼和斑头六线鱼视锥细胞密度最高处分别位于视网膜的腹-颞(V-T)区、腹区(V)、腹-颞(V-T)区和背-颞区(D-T),与各自的生活习性和摄食行为相适应。(4)4种岩礁性鱼类的成熟个体中,铠平鲉的最小分辨角最大,其次为花斑平鲉和大泷六线鱼,斑头六线鱼的最小分辨角最小。分析认为,即使是同样栖息于礁区的鱼类,其视觉特征也存在差异,这或许可以为岩礁区选择性渔具的开发提供一些有益的参考。     

Composition in essential and non-essential elements of early stages of cephalopods and dietary effects on the elemental profiles of Octopus vulgaris paralarvae

During the present study, we aimed at providing a first look at the elemental composition of the early stages of cephalopods as an approach to their elemental requirements in culture. Essential and non-essential elemental profiles of the European cuttlefish Sepia officinalis, the European squid Loligo vulgaris and the common octopus Octopus vulgaris laboratory hatchlings and wild juveniles were analysed. In addition, for O. vulgaris we determined elemental profiles of mature ovary, eggs in different stages of development and followed possible effects of four dietary treatments during paralarval rearing, also analyzing elemental content of the live preys Artemia nauplii and Maja brachydactyla hatchling zoeae. Content was determined for essential (As, Ca, Cr, Co, Cu, Fe, K, Mg, Mn, Na, Ni, P, Rb, S, Sr, Zn) and non-essential (Ag, Al, Ba, Cd, Hg, Pb) elements. The content in non-essential elements found in hatchlings and juveniles of the three species analyzed here seems to be far lower in comparison with subadult and adult stages of coastal cephalopods. In the octopus eggs, the non-essential element concentrations remained globally low compared to hatchlings and juveniles indicating the absorption of these elements along the ontogenetic development. The elemental composition of the octopus ovary and of the eggs, hatchlings and juveniles of the three cephalopod species analyzed here showed a high content in S. As expected, the calcified internal shell of the cuttlefish, rich in Ca and Sr, originates the main difference between species. It is remarkable the richness in Cu of hatchling octopus, that may indicate a particular nutritional requirement for this element during the planktonic life. The reared octopus paralarvae feed on Artemia nauplii, a prey with relatively low Cu content, showed nearly half Cu content that the “natural” profile of octopus hatchlings or wild juveniles. This suggests a dietary effect and/or an indication of the poor physiological stage of the Artemia-fed paralarvae. At the present, the percentage of essential element absorption by food or seawater is unknown for cephalopods and should be determined in the future to understand their feeding requirements in culture.     

罗非鱼海豚链球菌病的病理学观察

采用光学显微镜和透射电子显微镜技术分别对患海豚链球菌病的罗非鱼的病理学变化进行了研究。分离的病原菌革兰氏染色呈阳性,透射电镜负染观察菌体球形或卵圆形,直径0.6~1.0μm,多数呈链状排列。组织学病变主要表现为全身多组织、器官水肿,出血、变性、坏死以及炎症反应,特别是肝、脾、肾和脑分别表现为肝炎,脾炎,间质性肾炎和脑膜炎。超微结构观察发现病鱼肝、脾、肾、脑、心肌和骨骼肌等器官的细胞超微结构都有较为严重的破坏,细胞核畸形,染色质浓缩或边集,粗面内质网囊泡化及脱粒,线粒体肿胀,嵴断裂或溶解消失。研究表明,海豚链球菌能造成罗非鱼全身性组织器官病变,致使器官功能障碍,正常生理代谢调节紊乱,最后导致死亡。     

半滑舌鳎StAR基因克隆及其在不同组织的时空表达分析

利用半滑舌鳎性腺转录组测序获得的StAR基因部分序列,设计RACE引物,克隆了半滑舌鳎StAR基因的cDNA序列,全长为1 294 bp,5'端UTR为132 bp,3'端UTR为310 bp,开放阅读框(ORF)为852 bp,共编码283个氨基酸。将半滑舌鳎StAR基因与其他物种StAR基因进行氨基酸同源性分析,结果显示,半滑舌鳎StAR与塞内加尔鳎、大口黑鲈、花鲈、金头鲷的同源性都达到了85%,与虹鳟、斜带石斑鱼及日本鳗鲡的同源性分别为81%、83%和76%。雌、雄鱼不同组织StAR基因的表达分析表明,StAR基因在雄鱼性腺中高表达,在雄鱼的肝脏、脑及心脏中表达量较低,而在雄鱼的其他组织中不表达;在雌鱼肠中不表达,在其他组织(卵巢、肝脏、脾脏、脑、垂体、肌肉、心脏、肾脏)中微量表达。荧光定量PCR分析不同组织与不同时期性腺表达谱表明,雄鱼性腺中StAR基因的表达量显著高于雌、雄鱼其他各组织(P0.05),提示StAR基因对雄鱼精巢发育起重要作用。雄鱼不同时期表达谱分析结果显示,StAR基因在66天前的精巢中不表达,在150天时表达量急剧增加,至2龄时表达量最高,3龄时表达量下降,说明该基因在精巢发育成熟过程中起重要作用。原位杂交结果显示,StAR基因主要在雄鱼精巢的精子细胞中表达,而在雌鱼的卵巢中不表达。研究表明,StAR基因在半滑舌鳎精巢发育中发挥作用,且可能在精子形成中发挥重要作用。     

Distribution of a fish pathogen Listonella anguillarum in the Japanese flounder Paralichthys olivaceus hatchery

The present study was undertaken to investigate the distribution of Listonella anguillarum in a Japanese flounder (Paralichthys olivaceus) hatchery. A total of 2704 isolates were obtained from the developing fish, live diets and artificial feeds of Japanese flounder and their rearing water, 439 of which were identified as L. anguillarum by the combining incubation on thiosulfate-citrate-bile salt-sucrose (TCBS) agar at 35 °C overnight with polymerase chain reaction (PCR) detection for the VAH1 hemolysin gene. L. anguillarum was detected in all seven rotifer samples, with densities of 2.5 × 10³ to 4.6 × 10⁶ colony forming units (CFU) g^− 1. Both the analyzed samples of Nannochloropsis oculata contained this bacterium at densities of 1.6 × 10⁴ to 1.4 × 10⁵ CFU g^− 1. L. anguillarum was detected in only one of four samples of Artemia nauplii with a density of 4.8 × 10⁵ CFU g^− 1 (35%) and it was not detected in the two analyzed artificial feed samples. L. anguillarum was detected in 11 of 18 specimens of larval and juvenile Japanese flounder at densities of 5.0 × 10¹ to 7.4 × 10⁵ CFU g^− 1, while it was not detected in the two analyzed egg specimens of Japanese flounder. These results indicate that L. anguillarum associated with the developing Japanese flounder is likely derived from rearing water and live diets such as rotifers. Further, it is strongly suggested that L. anguillarum is a transient bacterium of the intestinal microflora for the Japanese flounder but is a permanently indigenous one for the Japanese flounder hatcheries.