低盐胁迫下LXRα激动剂添加和RNA干扰对大菱鲆(Scophthalmus maximus)幼鱼脂质代谢的影响

大菱鲆是我国重要的养殖经济鱼类,随着养殖水域逐渐向滩涂和内陆拓展,亟待研究其低盐适应性机制,进而为培育耐低盐良种提供参考。通过腹腔注射干扰剂dsRNA以及激动剂T091317,以进一步检测下游脂质代谢相关基因的表达以及血清脂代谢产物的变化。研究设置两个盐度梯度(5和30),每个盐度设置4个处理组(干扰组、激动组、生理盐水组和空白对照组),实验周期72 h。结果显示:在dsRNA注射后,肝脏中LXRα基因的相对表达量呈现出显著的抑制作用,尤其是在盐度30下;盐度5的肝脏组以及不同盐度的肠道组中,干扰效果不明显。激动剂注射后LXRα基因的相对表达量在不同盐度组中均呈现了激动效应,但维持时间较短。低盐可以抑制SREBP-1、CYP7A1和ApoA-IV基因的表达,当LXRα基因受到抑制或激动后,三者尽管表达趋势不同,但整体上均呈现出相应的干扰和激动效应。血清生化结果显示,低盐显著抑制了TG和HDL的含量,对T-CHO以及LDL的含量无显著影响。30盐度下,当LXRα基因受到抑制或激动后,TG的含量呈现出相应的干扰和激动效应。以上表明在腹腔注射干扰剂dsRNA以及激动剂后,都能够相应的导致LXRα基因表达量降低或者升高。LXRα基因受到抑制后,可以引起下游转录因子SREBP-1基因表达的降低,进而抑制了TG的合成,因此血清中甘油三酯的含量可以进一步作为大菱鲆耐低盐选育的表型。

Transcriptome analysis reveals immune-related genes in tissues of Vibrio anguillarum-infected turbot Scophthalmus maximus

Turbot Scophthalmus maximus is an important mariculture fish species with high economic value.However,the bacterial diseases caused by Vibrio anguillarum infection bring huge economic losses to the turbot aquaculture industry.To understand the immune response of the turbot against V.anguillarum infection and to explore novel immune-related genes,the transcriptome analysis of turbot spleen and gills were conducted after V.anguillarum infection.Differentially expressed genes(DEGs)were identified in spleen and gill of the turbot amounted to 17261 and 16436,respectively.A large number of immunerelated DEGs were enriched in cytokine-cytokine receptor interaction signaling pathway,and the others by the kyoto encyclopedia of genes and genomes(KEGG)enrichment.The gene ontology(GO)classification analysis revealed that V.anguillarum infection had the greatest effect on biological processes and cellular components.Twelve immune-related DEGs were identified in the spleen(cstl.1,egfl6,lamb21,v2rx4,calcr,and gpr78a)and gills(ghra,sh3gl2a,cst12,inhbaa,cxcl8,and il-1b)by heat map.The proteinprotein interaction(PPI)networks were constructed to analyze the immune mechanism.The results demonstrate that the maturation and antigen processing of major histocompatibility complex(MHC)class II molecule,and calcitonin-or adrenomedullin-regulated physiological activity were important events in the immunity of turbot against V.anguillarum infection.In the gills,the protein interactions in TGF-βsignaling pathway,production of inflammatory factors,and endocytosis regulation were most significant.Our research laid a foundation for discovering novel immune-related genes and enriching the knowledge of immune mechanisms of turbot against V.anguillarum infection.

Establishment,Characterization and Expression Pattern of a Spleen Cell Line,SMSP,Derived from Turbot(Scophthalmus maximus)

A turbot(Scophthalmus maximus)cell line named SMSP was obtained from the spleen.The origin of the cells was identified by morphology,chromosome number and COI gene.The optimal basic medium,serum concentration and growth temperature of the cells were detected.SMSP cell line is mainly composed of fibroblast-like cells.Most of the SMSP cells contained 44 chromosomes,and the sequence of COI gene confirmed that the cells were originated from turbot.The optimal culture conditions were 24℃,DMEM+10%FBS.The cell line had high transfection efficiency for siRNA and plasmid.After stimulation with lipopolysaccharide(LPS)or poly(I:C),the expressions of immune-related genes such as TNF-β,IL-12s,IL-10 and IL-1βwere up-regulated significantly in the early stage(P<0.05).This study will provide a model for exploring immune mechanism of turbot against pathogen in vitro.

The ontogeny of acidic digestive tract in larval turbot Scophthalmus maximus based on H+/K+-ATPase and pepsinogen

Acidic digestion is an important digestive process of marine fish.In fish stomach,two enzymes are involved in the secretion of hydrochloric acid(HCl)and proteomic digestion:H^(+)/K^(+)-ATPase and pepsinogen.However,the starting of digestive function in fish is still unclear.To reveal the details of acidic digestion of turbot Scophthalmus maximus in early development,a 40 day of turbot larvae culture was conducted.The H^(+)/K^(+)-ATPase gene from the turbot S.maximus(smH^(+)/K^(+)-ATPase)was identified and characterized.Based on our previous discription on pepsinogen of turbot S.maximus,we combined pepsinogen and H^(+)/K^(+)-ATPase and analyzed the mechanism of acidic digestion in turbot.Results show that the spatial and temporal expression profiles of H^(+)/K^(+)-ATPase agreed with pepsinogen A and C in turbot,indicating a synergetic action between H^(+)/K^(+)-ATPase and pepsinogen during the acidic digestion process.In addition,the turbot juveniles showed a faster growth after the expressions of H^(+)/K^(+)-ATPase gene and pepsinogen gene,demonstrating that pepsin had a higher digestive efficiency,for which a compound diet should be provided to the fish from Day 22 onward.This study provided a reference for biology research and aquaculture of turbot and other marine fishes.

大菱鲆(Scophthalmus maximus)一种CXC趋化因子的克隆及其表达特征

从大菱鲆脾脏cDNA文库中筛选到一种CXC趋化因子。它的全长cDNA含有一个81bp的5’UTR,一个414bp的开放阅读框和一个449bp的3’UTR。开放阅读框编码了137个氨基酸残基。经过PCR扩增发现,该CXC趋化因子基因内含有4个外显子和3个内含子。RT-PCR分析表明,大菱鲆CXC趋化因子在正常脾脏和头肾中表达强烈;在胚胎的不同发育时期,大菱鲆CXC趋化因子从体节期开始才有微弱的表达。大菱鲆胚胎细胞(TEC)在用鳗弧菌感染后6h也有强烈的表达。这些结果表明该CXC趋化因子在大菱鲆免疫应答中起着重要作用。

Interaction of temperature and salinity on the expression of immunity factors in different tissues of juvenile turbot Scophthalmus maximus based on response surface methodology

Central Composite Design(CCD) and response surface methodology were used in the experiment to examine the combined effect of temperature(16-28℃) and salinity(18-42) on Hsp70 and IgM genes expression levels in turbot(Scophthalmus maximus) liver and kidney.The results showed that the coefficients of determination(R^2=0.965 2 for liver Hsp70,0.972 9 for kidney Hsp70,0.921 for liver IgM and 0.962 1 for kidney IgM) and probability values(P<0.01) were significant for the regression model.The interactive effect between temperature and salinity on liver Hsp70,kidney Hsp70 and liver IgM were not significant(P>0.05),while the interactive effect between temperature and salinity on kidney IgM was significant(P<0.01).The model equation could be used in practice for forecasting Hsp70 and IgM genes expression levels in the liver and kidney of juvenile turbot via applying statistical optimization of the response of interest,at which the maximum liver Hsp70,kidney Hsp70,liver IgM and kidney IgM of1.48,1.49,2.48,and 1.38,respectively,were reached.The present model may be valuable in assessing the feasibility of turbot farming at different geographic locations and,furthermore,could be a useful reference for scientists studying the immunity of turbot.

Preliminary Study of 12 Chinese Herbs as Feed Attractants on Turbot Juvenile(Scophthalmus maximus)

[Objective] The paper aims to study the attractive effects of 12 Chinese medicinal herbs on Scophthalmus maximus juvenile.[Method] We took 200 Scophthalmus maximus with the average body length of 3 cm,average weight of 1 g and 60 days of age as the research object,and 12 Chinese medicinal herbs as feed attractants were selected,including Cloves,Cinnamon,Star anise,Kaempferol,Mint,Coptis,Phellodendron,skullcap,Gardenia,Sophora flavescens,Mume,Myrobalan meat were selected.The amount added to the basic diet was 2‰..The times of turbot juveniles going into each inducing room and the times of turbot juveniles biting the feed ball were recorded by maze experiments.Inducing effects of the Chinese herbal medicines were analyzed.[Result] The results showed that the attractant effect of Mume,Gardenia,Scutellaria and kaempferol had the significantly inducing effect on Scophthalmus maximus juvenile.And 2‰ Gardenia,5‰ Matrine,5‰ Scutellaria and 2‰ Kaempferol have the best attractant effects.In compound experients,the attractive effect of mixture of 2‰ matrine and gardenia on Scophthalmus maximus was the best.[Conclusion] The attractive effect on Scophthalmus maximus has something to do with the kind and concentration of Chinese medicinal herbs.

Turbot Scophthalmus maximus: stocking density on growth, pigmentation and feed conversion

Juvenile turbot （Scophthalmus maximus） the initial stock densities at 0.28, 0.87, 1.12, 1.16, 2.75 were reared in five different experiment groups in kg/m^2 respectively for the study of the density effect on growth, pigmentation and feed coefficient rate. The experiment lasted for 60 days with final stock densities at 1.91, 6.31, 8.86, 11.97, 17.67 kg/m^2 respectively. Result showed that, in the same experiment condition, the stocking density has a positive effect on growth in low density and negative in high density. The SGR （special growth rate） was 3.189, 3.304, 3.447, 3.341, and 3.087 respectively. The uniformity of weight distributing among experiment groups decreased with increasing density. The stocking density had positive effect on feed coefficient rate. Group 1 had the least feed coefficient rate 0.96, and highest at 1.25 in Group 5, the highest density group. High stocking density inhibited the growth and increased the feed coefficient rate. The stocking density had negative relationship to pigmentation improvement for whitened fish.

不同铁水平饲养对大菱鲆(Scophthalmus maximus)生长及耐低氧能力影响的研究

铁元素作为细胞色素的重要成分参与鱼类细胞内的氧化还原供能过程,在鱼类应对低氧胁迫时发挥重要作用。以硫酸亚铁为铁源,研究不同铁水平饲养对大菱鲆(Scophthalmus maximus)生长及耐低氧能力的影响。在含铁量为463 mg/kg的基础饲料中添加0(A组)、75(B组)、150(C组)、225(D组)和300(E组)mg/kg铁离子,探究不同铁水平饲喂对大菱鲆常氧下的生长、血清生理生化指标以及抗低氧胁迫的影响。结果显示,常氧情况下,随着饲料中铁含量的增加,上述指标呈先增加后下降的趋势(P<0.05),且C、D组均为峰值显著高于A、B、E三组(P<0.05)。低氧胁迫及耐受实验结果显示,随着低氧胁迫的加剧,C、D两组抗氧化酶活性(SOD,GSH-PX)、低氧[DO=(2.0±0.5)mg/L]死亡率均显著高于或低于其他三组(P<0.05)。铁作为重要的氧载体和电子传递体影响着生物体的物质代谢和能量代谢。研究表明饲料铁水平613~688 mg/kg的饲养对大菱鲆生长性能、抗氧化系统的增强和提高低氧耐受能力具有促进作用,研究结果可为鱼类健康养殖和功能性饵料研制提供新数据支撑。

Pharmacokinetics and tissue behavior of enrofloxacin and its metabolite ciprofloxacin in turbot Scophthalmus maximus at two water temperatures

Turbot Scophthalmus maximus, an important aquaculture species in China, currently suffers from epizootic diseases because of high density aquaculture. Enrofloxacin has been used to treat various systemic bacterial fish infections. However, studies concerning the pharmacokinetics of enrofloxacin in turbot are limited. In this study, the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin, were investigated in the turbot following intravenous and oral administration at 10 mg enrofloxacin/kg body weight, at 16℃ and 10℃ water temperatures. The concentrations of enrofloxacin and ciprofloxacin in the main tissues （plasma, muscle, liver and kidney） were detected by HPLC. The results show that the plasma concentration-time data for enrofloxacin were best described as a two-compartment open model after intravenous and oral administration. Three pharmacokinetic equations were established between the concentrations and temperatures. The kinetic profile of enrofloxacin was temperature dependent. The absorption half-life of enrofloxacin was 1.99 h and 2.17 h after oral administration, whereas the elimination half-life of the drug was 98.63 h and 136.59 h at 16℃ and 10℃, respectively. The peak concentration of enrofloxacin in plasma and tissues was higher at 16℃ than that at 10℃, and the peak plasma concentration time in the liver was the shortest at both temperatures among those of other tissues. The plasma ℃/MIC ratio varied between 11.08 and 5 540.00 at 16℃; and between 7.92 and 3 960.00 at 10℃. The AUC/MIC ratio was 467.82-280 690.00 at 16℃, and 359.48-215 690.00 at 10℃. These ratios indicate that it is possible to obtain therapeutic efficacy. Very low levels of ciprofloxacin were detected. The AUC ratios of ciprofloxacin and enrofloxacin in plasma suggest that plasma ciprofloxacin might play a minor role in enrofloxacin treatment for turbot.

Protein-sparing effect of carbohydrate in diets for juvenile turbot Scophthalmus maximus reared at different salinities

The aim of the present study was to investigate the protein-sparing effect of carbohydrate in diets for juvenile turbot(Scophthalmus maximus) reared at five salinities(12,18,24,30,and 36).The fish were fed three isocaloric and isolipidic diets for 60 days.The results show that specific growth rate(SGR)and feed conversion efficiency(FCE) were higher in fish reared at salinities of 18 and 36,but lower at 12.Fish fed with diet C25P40(25%carbohydrate and 40%protein) had lower SGR and FCE values compared with those fed with the C5P52(5%carbohydrate and 52%protein) and C15P46(15%carbohydrate and 46%protein) diets;however,there was no statistical difference between diet C5P52 and C15P46.SGR and FCE values were unaffected by diet composition in fish reared at salinity 36.Hepatic lipogenic enzyme activities were higher in fish reared at 18 and 36,but lower at 12,while glucokinase(GK) activity was higher in fish reared at 12,and lower at 18 and 36.Dietary starch enhanced GK activity while depressing lipogenic enzyme activity.However,lipogenic enzyme activity increased with increasing dietary starch in fish reared at 36.It is recommended that salinity should be maintained > 12 in the farming of juvenile turbot.In addition,an increase in gelatinized starch from 5%to 15%could spare 6%dietary protein in fish reared at salinities of 18-30,while higher salinity(36) could improve dietary carbohydrate use and enhance the protein-sparing effect,which is linked with the induction of lipogenic capacities.

Heritability and genetic correlation of survival in turbot(Scophthalmus maximus)

We analyzed the survival data of the offspring from 21 sires and 42 dams of turbot. The results show that the cumulative survival rates for turbot from 2 to 18 months range from 17.5% to 28.5% main mortality occurred during months 2-5; and the highest survival rates of families were 97.9%, 98.8%, 99.4%, 99.7% during months 2-5, 5-6, 6-8, 8-11, and ll-18, respectively, and 99.5%, being 53.5%, 23.8%, 19.5%, 14.9%, and 13.2% higher, respectively, than the mean values in each period. In all periods, the estimated heritabilities for survival were very low without significant difference from zero （P〉0.05） （values ranged from 0.06 to 0.12）, indicating low additive genetic effects. The genetic correlations of survival among families in different periods were all positive, but low in magnitude （values range from 0.03 to 0.31）. Genetic correlations between long-term survival and other periods＇ survival had negative values （-0.06 and -0.15） and three positive values （0.16, 0.12 and 0.14）. Genetic correlations between survival and weight were all positive, except for survival at months 2-5 and weight at 18 months, which was not significantly negative （-0.18）.

Genetic parameter estimation for juvenile growth and upper thermal tolerance in turbot(Scophthalmus maximus Linnaeus)

Twenty-six half-sib groups （53 full-sib families） of turbot,Scophthalmus maximus Linnaeus, were obtained by artificial insemination. We measured growth in the offspring （40-50 individuals/family） and subjected them to a thermal tolerance challenge over a period of 34 d. There was no significant difference in daily mor-tality （range： 0.580%-1.391%） between Days 1-13 during the thermal tolerance challenge. However, daily cumulative mortality increased rapidly between Days 14 and 29, especially on Days 15 and 16 （20.232% and 34.377%, respectively）. Mortality was highest on Day 16 （14.145%）. We estimated the genetic parameters using the average information restricted maximum likelihood method. We used a likelihood ratio test to evaluate the significance of effects in models with and without identity as an effect, and compared the final log-likelihoods （maximum log L）. Lastly, we estimated phenotypic and genetic correlation between the up-per thermal tolerance limit （UTT） and body weight （BW）. In this study, the positive phenotypic correlation was low between UTT and BW （0.093±0.029）. The genetic correlation between UTT and BW was negative （-0.044±0.239）. The heritability for upper thermal tolerance was low （0.087±0.032）, which is of approximate-ly moderate heritability. The heritability for body weight was high （0.303±0.074）. Our results suggest there is significant potential for improvement in the culture of turbot by selective breeding.

大菱鲆(Scophthalmus maximus)热休克蛋白SmHsp47相互作用蛋白His-pull down和质谱鉴定

首次克隆了大菱鲆(Scophthalmus maximus)热休克蛋白SmHsp47基因。该基因cDNA序列全长为1927 bp,其中开放阅读框长度为1218 bp,编码一条长度为405个氨基酸的多肽链。结果表明,大菱鲆SmHsp47基因在肝脏、皮肤、鳃和肠等4个组织中都有表达,表达量最高的组织是肝脏,表达量最低组织是鳃;25℃处理6 h后SmHsp47在皮肤中的表达量增加150多倍。利用大肠杆菌原核表达系统表达了SmHsp47的his标签融合蛋白,然后利用His-pull down技术捕获了SmHsp47的相互作用蛋白质,通过质谱分析鉴定出31种候选蛋白,其中大部分蛋白为参与翻译后修饰,蛋白转换及分子伴侣;此外还包括参与脂转运与代谢、能量产生与转换、细胞骨架、防御机制等过程的蛋白质。从31种候选蛋白中进一步筛选出3种分值较高的蛋白:未知蛋白(uncharacterized protein,A0A6A4TFV0)、胎球蛋白B(Fetuin B,A0A2U9C388)和胶原结合蛋白(collagen-binding protein,A0A2U9B608),可为后续的研究提供方向。

Developmental quantitative genetic analysis of body weights and morphological traits in the turbot, Scophthalmus maximus

In order to elucidate the genetic mechanism of growth traits in turbot during ontogeny, developmental genetic analysis of the body weights, total lengths, standard lengths and body heights of turbots was conducted by mixed genetic models with additive-dominance effects, based on complete diallel crosses with four different strains of Scophthalmus maximus from Denmark, Norway, Britain, and France. Unconditional genetic analysis revealed that the unconditional additive effects for the four traits were more significant than unconditional dominance effects, meanwhile, the alternative expressions were also observed between the additive and dominant effects for body weights, total lengths and standard lengths. Conditional analysis showed that the developmental periods with active gene expression for body weights, total lengths, standard lengths and body heights were 15–18, 15 and 21–24, 15 and 24, and 21 and 27 months of age, respectively. The proportions of unconditional/conditional variances indicated that the narrow-sense heritabilities of body weights, total lengths and standard lengths were all increased systematically. The accumulative effects of genes controlling the four quantitative traits were mainly additive effects, suggesting that the selection is more efficient for the genetic improvement of turbots. The conditional genetic procedure is a useful tool to understand the expression of genes controlling developmental quantitative traits at a specific developmental period（t-1→t） during ontogeny. It is also important to determine the appropriate developmental period（t-1→t） for trait measurement in developmental quantitative genetic analysis in fish.

Effects of probiotic on microfloral structure of live feed used in larval breeding of turbot Scophthalmus maximus

The effects of an exogenous probiotic( Bacillus amyloliquefaciens) on microbial community structure of B ranchionus plicatils and A rtemia sinica were evaluated in this study during turbot( Scophthalmus maximus) larval breeding. The analysis and comparison of the microfloral composition of live feed with probiotic was conducted using the Illumina HiSeq PE250. The abundance of microbial species and diversity of microflora in live feed with B. amyloliquefaciens were higher than those in the control. The microfloral composition was similar among the three replicate experimental groups of B. plicatils compared with the control after enrichment. L actococcus, Pseudoalteromonas, and Alteromonas were always dominant. Additionally, some other bacterial species became dominant during the enrichment process. The microbial community during nutrient enrichment of A. sinica was rather similar among the three control replicates. Relative abundance of Cobetia sp., the most dominant species, was 54%–65.2%. Similarity in the microbial community was still high after adding B. amyloliquefaciens. Furthermore, P seudoalteromonas and Alteromonas replaced Cobetia as the dominant species, and the abundance of Cobetia decreased to 4.3%–25.3%. Mean common ratios at the operational taxonomic unit level were 50%–60% between the two B. plicatils and A. sinica treatments. Therefore, the microbial community structure changed after adding B. amyloliquefaciens during nutrient enrichment of B. plicatils or A. sinica and tended to stabilize. Additionally, the abundance of V ibrio in any kind of live feed was not significantly dif ferent from that in the control. These results will help improve the microflora of B. plicatils and A. sinica and can be used to understand the multiple-level transfer role of probiotic species among probiotic products, microflora of live feed, and fish larvae.

Isolation and characterization of 45 polymorphic microsatellite loci of turbot(Scophthalmus maximus)and cross-species amplification

Turbot （Scophthalmus maximus） is a flatfish species commercially important for aquaculture. In this study, we generated a microsatellite-enriched genomic DNA library for Scophthalmus rnaxirnus, and then isolated and characterized 45 microsateIIite loci by genotyping 30 individuals. The observed number of alleles ranged from 2 to 19 with an average of 6.24, while the effective number of alleles ranged from 1.30 to 11.11 with an average of 3.66. The expected heterozygosities varied from 0.235 to 0.9254 and polymorphic information content ranged from 0.204 4 to 0.903 3, with an average of 0.622. Twelve loci deviated significantly from Hardy-Weinberg equilibrium, and no significant linkage disequilibrium was observed between any pair of loci after Bonferroni correction. In cross-species amplification, five flatfish species （Paralichthys lethostigma, Verasper rnoseri, platichthys stellatus, Hippoglossoides dubius and Cynoglossus semilaevis） showed at least one polymorphic locus. These polymorphic microsatellite loci should prove useful for population analysis of turbot and other related species.

Estimates of heritability and correlation for growth traits of Turbot(Scophthalmus maximus L.) under low temperature conditions

The objectives of this present research were to assess the heritability of growth traits under low temperature conditions in turbot（Scophthalmus maximus L.）, and to analyze the correlation between body weight（BW） and body length（BL）. There were 536 individuals from 25 full- and half-sib families involved in this study. During the entire 90-day period, which was initiated at 233 dph（day old） and ended at 323 dph, the individuals＇ BW and BL were weighed consecutively six times every 18 days. The heritability of BW and BL and the correlation between these two traits were estimated based on an individual animal model with the derivative-free restricted maximum likelihood（DFREML） method. These results showed that the specific growth rates（SGR） of 25 families was from 0.75±0.11 to 1.05±0.14 under water temperature of 10.5–12°C. In addition, the heritability of BW and BL estimated under low-temperature were 0.32±0.04 and 0.47±0.06, respectively. The BW had a medium heritability（0.2–0.4）, and the BL had a high heritability（〉0.45）, which suggested that selection for increased weight and length was feasible. Moreover, there was potential for mass selection on growth. The genetic and phenotypic correlations between BW and BL were 0.95±0.01 and 0.91±0.01（P 〈 0.01）, respectively. A significant correlation between BW and BL showed that BL could be instead of BW for indirect selection, which could be effectively implemented in the breeding program.

Morphological comparison between a selected fast-growing strain and the common cultured strain of turbot Scophthalmus maximus

To evaluate the effect of genetic improvement of the turbot Scophthalmus maximus, we analyzed morphological differences between a fast-growing strain obtained by family selection and the common cultured strain, by principal component analysis, stepwise discriminant analysis, and t-tests. Although they clearly differed morphologically, plots of the principal components of the two strains partially overlapped. However, the difference between the strains was supported with very high precision by discriminant analysis. The t-tests revealed that 4 of the 13 morphological traits analyzed were highly significantly different (P<0.01), 4 traits also differed significantly (P<0.05), and the remainder did not differ significantly. The coefficients of difference of the 13 traits were all lower than the threshold value between subspecies (1.28). Together, the results indicate that a trend for segregation of characters from the common cultured strain have already appeared in the selected fast-growing strain but the degree of segregation have not risen to subspecies level.

Ciliate Uronema marinum is the causative agent of scuticociliatosis in farm raised turbot Scophthalmus maximus

To identify the pathogen that causes scuticociliatosis in farmed-raised turbot Scophthalmus maximus , we isolated a ciliate from the brain tissue of an infected turbot and identifi ed it as Uronema marinum based on morphological and molecular evidence. We then infected the turbots in artifi cial laboratory settings with pure cultured U . marinum . The infected turbots showed syndromes similar to those observed in naturally infected ones. Furthermore, microscopic examination and PCR detection confi rmed the presence of the ciliate in the tissues of those laboratory-infected turbots. To our best knowledge, this is the fi rst report of scuticociliatosis caused by U . marinum in farm-raised turbot S . maximus in China.