虹鳟源嗜冷黄杆菌的分离鉴定及致病性

为确定患病虹鳟的病原,本实验从患病鱼溃烂肌肉中分离到2株细菌,分别命名为CH06和CH07,经回归感染证实分离菌株为导致此次虹鳟患病的病原菌,并进一步对其形态特征、理化特性、分子特征、血清型及耐药性进行分析.结果显示,CH06和CH07株在TYES琼脂平板上呈煎蛋状外观,产黄色素,氧化酶和过氧化氢酶呈阳性,能水解明胶和...     

虹鳟源嗜冷黄杆菌的分离鉴定及致病性研究

为确定患病虹鳟的病原,本实验从患病鱼溃烂肌肉中分离到2株细菌,分别命名为CH06和CH07,经回归感染证实分离菌株为导致此次虹鳟患病的病原菌,并进一步对其形态特征、理化特性、分子特征、血清型及耐药性进行分析。结果显示,CH06和CH07株在TYES琼脂平板上呈煎蛋状外观,产黄色素,氧化酶和过氧化氢酶呈阳性,能水解明胶和酪蛋白,不能水解淀粉,不能利用果糖、半乳糖和七叶苷等。16S rRNA比对结果显示,CH06和CH07株与嗜冷黄杆菌模式株NBRC 15942的同源性分别为99.35%和99.42%。综合菌株理化和分子特性确定CH06和CH07株为嗜冷黄杆菌。利用多重PCR方法鉴定CH06和CH07株的血清型均为1型(Fd型);多位点序列分型(MLST)分析表明,CH06和CH07株的基因型分别为ST-12和ST-78型,且均属于CC-ST10克隆型。人工感染结果显示,CH06和CH07株对虹鳟幼鱼具有较高致病性,其半致死浓度(LD₅₀)分别为7.1×10⁵和1.1×10⁵ CFU/mL,攻毒剂量与临床病症出现时间呈反比,从人工感染实验鱼的肌肉、脾脏等组织中可重新分离到嗜冷黄杆菌。组织病理变化显示,病鱼肝细胞肿胀,空泡变性,部分肝细胞溶解坏死,细胞核溶解消失;脾脏充血、出血,淋巴细胞减少,红细胞和含铁血黄素增多;肌纤维间隙增宽、断裂、弯曲不齐,部分肌细胞肌浆溶解呈蜂窝状。CH06和CH07株对10种抗菌药物的耐药谱略有不同,均对氨苄西林和甲氧苄啶-磺胺甲噁唑敏感;CH06株对恩诺沙星、氟苯尼考等耐药,而CH07株对恩诺沙星和氟苯尼考中度敏感。本研究首次报道了我国虹鳟源嗜冷黄杆菌的分离鉴定及生物学特性,以期为虹鳟细菌性冷水病的防控提供科学依据。     

肌肉注射感染嗜冷黄杆菌虹鳟的组织嗜性与动态分布

嗜冷黄杆菌(Flavobacterium psychrophilum)是鲑鳟类细菌性冷水病(bacterial cold water disease, BCWD)的病原菌, 该病的发生和流行严重制约了鲑鳟产业的健康发展。本研究分析嗜冷黄杆菌肌肉注射感染虹鳟(Oncorhynchus mykiss)后病原菌的动态组织分布情况, 以期为 BCWD 的防控提供理论支撑。以 1.0×108 CFU/mL 浓度的嗜冷黄杆菌 CH06 株菌液肌肉注射感染实验虹鳟, 感染后 12 h、24 h、96 h 观察虹鳟临床症状及组织病变, 并利用 qPCR Taqman 探针法检测各组织病原载量。患病鱼的临床病征表现为体色发黑、游动缓慢或不动、食欲不振, 尾柄部注射处肌肉溃烂, 鳃苍白, 脾脏肿大, 伴有腹水。组织病理观察显示, 嗜冷黄杆菌感染后实验鱼注射处肌纤维断裂、溶解; 脾窦扩张, 其内充满红细胞; 肾脏组织含铁血黄素增加, 出现大量空泡变性, 肾小管上皮细胞变性、坏死, 肾间质炎性细胞浸润。qPCR 检测结果表明, 肌肉注射感染 12 h 后即可在脾脏、肝脏、肾脏、肠道、鳃、注射处肌肉、脑和尾鳍中检出嗜冷黄杆菌, 注射处肌肉病原载量最高为(5.85±2.11)×10⁵ copy/μL。感染后 24 h, 脾脏、脑中病原载量较 12 h时上升最多, 其他组织病原载量与感染 12 h 时水平一致, 注射处肌肉病原载量为(6.48±2.07)×10⁵ copy/μL, 显著高于其他组织(P＜0.05)。感染 96 h 后脾脏中的病原载量显著高于其他组织(P＜0.05), 达到(1.15±0.58)× 10⁷ copy/μL; 肝脏、肾脏、脾脏中的病原载量较 24 h 时上升最多。所有被检组织中病原菌载量随时间延长均呈上升趋势。另外, 3 个时间点注射处肌肉的病原平均载量最高, 其次为脾脏, 然后是肾脏和鳃。嗜冷黄杆菌人工感染虹鳟后, 注射处肌肉、脾脏是细菌的重要增殖场所。综上, 嗜冷黄杆菌可随着血液循环进入虹鳟各组织, 并表现出对注射处肌肉、脾脏、肾脏和鳃较强的组织嗜性, 而对肝脏、尾鳍、肠道和脑组织的嗜性相对较弱; 感染时间和病变程度与组织中的病原载量呈正相关。     

患病异育银鲫中嗜冷黄杆菌的分离鉴定及组织病理学观察

为确定越冬期内江苏多地异育银鲫 (Carassiusauratus gibelio)暴发性死亡的原因,本研究利用高通量测序比较了健康样品和患病样品的微生物群落多样性和结构组成的差异,分析了异育银鲫病害暴发过程中的细菌类型及特性。结果表明：在属水平上,患病样品中黄杆菌属丰度最高。在种水平上,患病样品中嗜冷黄杆菌 (Flavobacterium psychrophilum)的丰度显著增加,分别达到63.01%和61.31%,显著高于健康组的1.55%（P < 0.05）。根据微生物群落特征分析结果,从患病样品体表的病灶处分离出优势病原菌NJ01,通过细菌形态学、生理生化分析、16S rDNA 序列比对确定 NJ01 菌株为嗜冷黄杆菌。人工感染NJ01菌株14d后,1.7×106～1.7×107 CFU/mL两组的死亡率达到100%,感染症状和自然发病症状一致。组织病理学观察发现,病鱼的肌细胞坏死,间质中充满了淋巴细胞;肝脏中细胞溶解坏死,细胞核消融;在脾脏中发现脾细胞散在坏死,伴随着充血的症状;肾小管上皮脱落,肾间质存在大量淋巴细胞。药物敏感试验结果表明,NJ01菌株对头孢西叮、头孢哌酮、庆大霉素和克拉霉素等敏感。本研究首次报道了嗜冷黄杆菌在异育银鲫中的致病性,这将为大宗淡水鱼“越冬综合征”的药物防治及其致病机理研究提供参考依据。     

传染性造血器官坏死病毒Sn1203株全基因组序列及系统进化分析

本课题组于2012年从中国东部的虹鳟鱼养殖场分离得到了严重威胁我国鲑鳟鱼养殖的传染性造血器官坏死病毒(IHNV)IHNV-Sn1203病毒株。根据GenBank中收录的IHNV病毒株的序列设计引物,将IHNV-Sn1203基因组序列分成5个片段进行RT-PCR克隆,最终得到完整的IHNV-Sn1203基因组序列。利用Lasergene和MEGA 5.0软件分析了IHNV-Sn1203株的全基因组序列、基因组编码蛋白、基因组末端和未翻译序列、以及病毒基因同源性和系统发育。结果发现,IHNV-Sn1203基因组全长11131nts,共编码6个病毒蛋白,大小分别为核蛋白(N)1176nts、磷蛋白(P)693nts、基质蛋白(M)588nts、表面糖蛋白(G)1527nts、聚合酶蛋白(L)5961nts和非结构蛋白(NV)336nts。IHNV-Sn1203株各基因间均具有保守的基因终止序列(Gene end,GE)、基因间序列(Intergenic regions,IG)、基因起始序列(Gene star,GS)及Kozak序列,以确保病毒基因的转录和翻译效率。系统进化分析发现,IHNV分为E、U、M、L和J共5个基因型,其中IHNV-Sn1203株与中国其他IHNV株具有显著的亲缘关系,属于J基因型中的J Nagano亚型。由G基因的进化分析发现,J基因型的病毒与U基因型亲缘关系最近,推测IHNV病毒最初可能通过进口鱼卵的方式由U基因型引入,随时间的推移逐渐进化为J基因型。     

鱼源致病菌皮特不动杆菌Ap-W20的全基因组序列及其毒力特征分析

皮特不动杆菌是一种新兴的鱼源致病菌,为了对该菌的致病机理进行研究并为该菌引起的鱼类疾病防治提供技术储备,实验选取皮特不动杆菌菌株Ap-W20进行了全基因组测序,并对其毒力特征进行了分析。结果发现,Ap-W20全基因组序列总长为4 399 705 bp,GC含量为38.78%,共预测到4 230个编码序列(CDS),存在4个质粒,GenBank数据库登录号为CP027658～CP027662。ANI分析结果证实,Ap-W20属于皮特不动杆菌,且ApW20与已知人源皮特不动杆菌AP₈82(96.69%)和环境源皮特不动杆菌PHEA-2(96.55%)相似度较高。Ap-W20与AP₈82、PHEA-2的比较基因组学分析发现,Ap-W20中的基因岛、前噬菌体和质粒数量最多,这很可能与它们各自的分离环境和致病力有关。通过与毒力因子数据库(VFDB)比对,在Ap-W20全基因组中预测到286个毒力基因,主要与黏附和抗吞噬等有关。Ap-W20中还存在Ⅰ、Ⅱ和Ⅵ型分泌系统、多套双组分调控系统以及细菌素合成基因簇等,表明鱼源皮特不动杆菌可能存在复杂的致病机制。...     

6株鱼源嗜水气单胞菌的分子鉴定与毒力基因分析检测

为了分子鉴定6株鱼源嗜水气单胞菌,并从分子层面验证通过检测毒力基因以推测嗜水气单胞菌潜在致病性的可行性。实验采用PCR扩增16 S rDNA和gyrB基因并结合系统发育树的构建和分析进行菌种的分子鉴定,检测气溶素（ aerolysin, aer）、溶血素（ haemoly-sin, hly）、丝氨酸蛋白酶（ serine protease, ahp）、热稳定细胞肠毒素（ heat-stable cytotonic enterotoxin, ast）和热敏感细胞肠毒素（ heat-labile cytotonic enterotoxin, alt）5种毒力基因,且使用Mega 5．2对核苷酸和氨基酸序列进行分析。结果显示6株菌均为嗜水气单胞菌Aero-monas hydrophila,检测出5种毒力基因中的至少4种,其中均检测出溶血素和2种肠毒素,序列分析表明气溶素、溶血素和丝氨酸蛋白酶的氨基酸序列高度保守。本研究基于16 S rD-NA和gyrB基因可以准确地对嗜水气单胞菌进行分子鉴定,6株菌的毒力基因丰富预示着一定的致病性, aer、 hly和ahp基因相对保守,编码的毒力因子高度同源,在临床分子诊断中建议使用aer、 ahp和hly基因对嗜水气单胞菌的潜在致病性进行检测。     

一株河鲈源致病性虫草菌Cordyceps confragosa CHL02菌株的全基因组测序及比较基因组分析

致病性虫草菌Cordyceps confragosa CHL02菌株是从患病河鲈(Perca fluviavilis)鱼体分离鉴定的一株昆虫致病菌,其无性阶段蜡蚧轮枝菌(Lecanicillium lecanii)广泛用于农业中昆虫防治.本研究基于IlluminaPE150测序平台进行CHL02菌株的全基因组测序,对测...     

贝莱斯芽孢杆菌LG37全基因组测序分析及无机氮代谢相关候选基因的筛选

前期研究发现贝莱斯芽孢杆菌(Bacillus velezensis) LG37可高效同化无机氮,但其机理尚不清楚。为解读其高效同化无机氮的机理,结合三代PacBio RS II和二代Illumina HiSeq 2000测序技术对贝莱斯芽孢杆菌LG37进行全基因组测序,在此基础上利用NR、KEGG、eggNOG、GO和CARD数据库进行序列注释、分析,并通过本地Blast+对无机氮代谢相关基因进行挖掘。测序结果表明：1)贝莱斯芽孢杆菌LG37的基因组为3 929 697 bp的环状染色体,GC含量为46.5%,包含3 854个蛋白质编码基因、86个tRNA基因和27个rRNA基因。2)共筛选出无机氮代谢相关候选基因94个,主要涉及编码感应蛋白、转录调控因子、转运蛋白、氧化还原酶和同化酶等,并对这些基因的GO功能进行了注释分析。综上,LG37全基因组测序及无机氮代谢相关基因的分析为芽孢杆菌降低养殖水体中无机氮的研究提供了基因水平数据,为芽孢杆菌微生态制剂降低水体中无机氮的应用研究提供了理论依据。     

一株河鲈源致病性虫草菌Cordyceps confragosa CHL02菌株的全基因组测序及比较基因组分析

致病性虫草菌Cordyceps confragosa CHL02菌株是从患病河鲈(Perca fluviavilis)鱼体分离鉴定的一株昆虫致病菌,其无性阶段蜡蚧轮枝菌(Lecanicillium lecanii)广泛用于农业中昆虫防治。本研究基于Illumina PE150测序平台进行CHL02菌株的全基因组测序,对测序数据进行组装和组分分析,进行基因预测与功能注释,预测次级代谢产物合成基因簇,并进行病原宿主互作以及比较基因组分析。测序结果显示,CHL02基因组大小为36.17 Mb,GC含量为53.09%;预测包含8093个编码基因、1618个转座因子(TEs)、4572个串联重复序列及114个tRNA;共注释7724个基因,其中,1985个基因获得KOG注释,GO聚类分析中,2687个基因参与代谢过程,预测到22个次级代谢产物合成基因簇,1162个基因参与病原宿主互作机制中。基因聚类分析和系统发育树均显示,CHL02菌株与参考菌株昆虫源粗糙虫草菌(C. confragosa) RCEF 1005具有较高的同源性。本研究首次报道了河鲈源致病性虫草菌C. confragosa CHL02菌株的全基因组序列并分析其基本特征,与参考菌株进行比较基因组分析,为后续深入开展该病菌侵染河鲈的作用机制等相关研究奠定理论基础。     

Immunization of rainbow trout, Oncorhynchus mykiss (Walbaum), with a low molecular mass fraction isolated from Flavobacterium psychrophilum

Flavobacterium psychrophilum, the causative agent of rainbow trout fry syndrome has become a widespread fish pathogen in freshwater aquaculture worldwide. In this study, a low molecular mass fraction (P25-33), with an approximate weight of 25-33 kDa, was identified among F. psychrophilum strains in an immunoblotting analysis with anti-F. psychrophilum sera. The immunogenic efficacy of the isolated and extracted P25-33 was investigated in two intraperitoneal immunization trials with rainbow trout, Oncorhynchus mykiss (Walbaum). The first trial included immunizations using P25-33 with Freund's complete adjuvant (FCA) and the second trial included immunizations using P25-33, formalin-inactivated whole and sonicated F. psychrophilum cell preparations without FCA. In both trials, antibody titres against F. psychrophilum were analysed with an enzyme-linked immunosorbent assay and the efficacy of the immunizations was determined by a challenge with F. psychrophilum. The P25-33 was shown to give rise to a protective immune response in rainbow trout after immunization with FCA, but not without FCA when a low concentration of P25-33 was used. Instead formalin-inactivated whole and sonicated cells of F. psychrophilum were able to protect the immunized fish more effectively when immunized without FCA. The results suggest that whole or sonicated F. psychrophilum cells could be better candidates for a cost-effective water-based injection vaccine than the immunogenic fraction.     

Passive immunization of rainbow trout,Oncorhynchus mykiss (Walbaum), against Flavobacterium psychrophilum,the causative agent of bacterial coldwater disease and rainbow trout fry syndrome

Flavobacterium psychrophilum, the causative agent of bacterial coldwater disease (CWD) and rainbow trout fry syndrome (RTFS), causes high mortality in cultured salmonids. The present study was designed to determine the role antibody plays in conferring protection to rainbow trout fry, Oncorhynchus mykiss (Walbaum), by passive immunization with convalescent serum or serum from adult rainbow trout immunized with F. psychrophilum, and goat anti-F. psychrophilum serum. In each experiment, rainbow trout fry were injected intraperitoneally with antiserum and challenged by subcutaneous injection with a virulent strain (CSF-259-93) of F. psychrophilum 24-h post-immunization. Relative percentage survival (RPS) ranged from 9-42% when rainbow trout fry (mean weight 1.3 g) were injected with a 1:2 dilution of 25 microL of convalescent serum ranging in enzyme-linked immunosorbent assay antibody titres from 1600-102400. Rainbow trout fry (mean weight 1.0 g) passively immunized with 25 microL of serum from immunized adult fish exhibited RPS values of up to 57%. In each of these experiments, RPS increased with increasing antibody titres against F. psychrophilum. Passive immunization with 25 or 50 microL goat anti-F. psychrophilum serum, however, did not confer protection to fry (mean weight 1.3 g). These results suggest that trout antibody plays a role in conferring protection to F. psychrophilum, but antibody alone is unable to provide complete protection.     

应用重复力模型估计虹鳟生长性状的遗传力和育种值

采用单性状重复观测值动物模型（重复力模型）对中国水产科学研究院黑龙江水产研究所渤海冷水性鱼类试验站5个品系后代的4026尾虹鳟 (Onchorynchus mykiss) 的生长性状（体重、体长和肥满度）进行了遗传参数和育种值的估计,并对各个性状的表型和遗传趋势进行预测。在此模型中考虑了年份-季节固定效应、父本固定效应和母本固定效应;随机效应有个体的加性效应和个体永久环境效应。结果表明：该5个品系虹鳟及后代体重的遗传力为0.35,体长为0.10、肥满度为0.34。个体间的育种值差异较大,仅排名前20位相差近10倍;从不同评定方法的秩相关来看,用综合育种值法与单个性状方法之间存在极显著的相关,其相关度分别为0.998, 0.877, 0.85. -0.071, -0.064和-0.13。用综合育种值对虹鳟个体的选择价值的评定和名次排列, 以及与单个性状的评定名次有着一定程度上的差别。体重、体长和肥满度性状,其表型趋势与遗传趋势基本一致的品系,均可通过表型值进行选择。     

Genetic diversity of Flavobacterium psychrophilum recovered from commercially raised rainbow trout, Oncorhynchus mykiss (Walbaum), and spawning coho salmon, O. kisutch (Walbaum)

Flavobacterium psychrophilum is the aetiological agent of rainbow trout fry syndrome and bacterial cold water disease. This study examined the genetic diversity of F. psychrophilum isolates retrieved from multiple epizootics at rainbow trout, Oncorhynchus mykiss, rearing facilities and from spawning coho salmon, O. kisutch. A total of 139 isolates were confirmed as F. psychrophilum by PCR assay and were further typed using pulsed-field gel electrophoresis (PFGE). Multiple epizootics at three proximally located rainbow trout rearing facilities were numerically dominated by three PFGE profiles, which accounted for 76% of all trout isolates. In coho salmon, 19 PFGE profiles were differentiated by PFGE and four numerically dominant PFGE profiles represented 56% of all coho salmon isolates. PFGE analysis also indicated that the average similarity of macrorestriction patterns of F. psychrophilum isolates was greater in rainbow trout than in coho salmon (88% vs. 70%). Furthermore, it was not unusual to isolate multiple PFGE profiles from a single coho salmon sample whereas only two PFGE profiles were shared between two sample dates separated by 1 month. It is clear that the domestic rainbow trout aquaculture facilities studied here were primarily affected by a complex of genetically related strains whereas spawning coho salmon supported a much more genetically diverse collection of F. psychrophilum.     

比较基因组学分析鱼源荧光假单胞菌的致腐和适应性

为探究荧光假单胞菌的强致腐和环境适应性,实验通过分析2株海水鱼源荧光假单胞菌的蛋白酶活性和挥发性盐基氮(TVB-N)的形成,并采用比较基因组学方法解析致腐和适应机制。结果显示,荧光假单胞菌PF07和PF08在冷藏鱼汁中的蛋白酶活性强,积累较多TVB-N。经全基因组测序、组装和功能注释后,得到PF07基因组长度为6.13 Mb,GC含量61.4%。通过泛基因组分析可知,PF07与PF08核心基因共4 980个,独特基因分别有516和470个。COG和KEGG注释显示2株致腐菌基因组中参与氨基酸代谢基因占比最高,PF07独特基因参与无机离子转运代谢居多。碳水化合物活性酶CAZy注释表明,2株荧光假单胞菌基因组中糖苷转移酶与糖苷水解酶基因均占比最高,还鉴定得到大量分解碳水化合物、蛋白质和脂质等多种底物的酶和相关蛋白基因,特别有碱性金属蛋白酶AprA、多胺ABC转运蛋白渗透酶PotC、精氨酸与鸟氨酸脱羧酶等多种降解蛋白酶。另外,2株致腐菌分布有rpoS、rpoN和rpoD多种σ因子。2株鱼源荧光假单胞菌表现出强的致腐性,研究通过比较基因组学方法解析荧光假单胞菌分布多种编码蛋白酶和腐胺形成的氨基...     

Effect of hydrogen peroxide and/or Flavobacterium psychrophilum on the gills of rainbow trout,Oncorhynchus mykiss (Walbaum)

The immune response and morphological changes in the gills of rainbow trout fry after immersion in hydrogen peroxide (H₂O₂), Flavobacterium psychrophilum or combined exposure were examined. The gills were sampled 4, 48, 125 and 192 h after exposure, and the regulation of expression of the following genes was investigated using qPCR: IgT, IgM, CD8, CD4, MHC I, MHC II, IL-4/13A, TcR-β, IL-10, IL-1β, IL-17, SAA and FoxP3. Bacteria were not observed in haematoxylin-and-eosin-stained gill tissue, but the presence of F. psychrophilum 16S rRNA was detected using qPCR. The 16S rRNA levels were correlated with gene expression. Although pretreatment with H₂O₂ before immersion in F. psychrophilum did not significantly alter the amount of bacteria found in the gill, the immune response was influenced: exposure to F. psychrophilum resulted in a negative correlation with expression of IL-17c1, MHC I and MHC II, while pretreatment with H₂O₂ resulted in a positive correlation with IL-4/13A and IgM. Exposure to either H₂O₂ or F. psychrophilum influenced the regulation of gene expression and damaged tissue. Exposure to both combined altered the immune response to infection and postponed healing of gill tissue.     

Phenotypic and genotypic characterization of Flavobacterium psychrophilum from Finnish fish farms

Finnish isolates ( n =37) of Flavobacterium psychrophilum isolated from farmed salmonids were studied using phenotypic and genotypic characteristics. The characteristics of isolates were compared with the characteristics of Swedish and Estonian F. psychrophilum isolates and the type strain, F. psychrophilum NCIMB 1947^T. Multiple isolates from eight disease outbreaks were examined to determine differences between isolates from a single outbreak. The F. psychrophilum isolates represented a biochemically homogeneous group. However, some minor differences in biochemical and physiological characteristics were observed. Seven different antigenic patterns among Finnish isolates were detected and the results suggest a new serotype of F. psychrophilum . Using Cla Ι, Hae ΙΙΙ and Pvu ΙΙ restriction enzymes in ribotyping analyses 13 different genotypes were demonstrated and a possible relationship between serotype Fd and genotype F1 was determined. There were no significant differences between the isolates from Finland, Estonia, Sweden and the type strain of F. psychrophilum .     

Detection of the bacterium Flavobacterium psychrophilum from a natural infection in rainbow trout, Oncorhynchus mykiss (Walbaum), using formalin-fixed, wax-embedded fish tissues

The bacterial pathogen Flavobacterium psychrophilum was successfully identified from formalin-fixed, wax-embedded tissue blocks of infected rainbow trout heart and spleen tissues, using a polymerase chain reaction (PCR)-based assay. Filamentous bacteria were observed in haematoxylin and eosin and Giemsa-stained sections but no bacteria were recovered from the diseased fish using standard bacteriology isolation techniques. All infected fish had histopathological evidence of myocarditis or rainbow trout fry syndrome. Immunohistochemistry was attempted using three different anti-F. psychrophilum sera but the results were inconclusive, and an alternative molecular approach was therefore attempted. This paper describes the use of a PCR-based assay to help identify bacteria present in formalin-fixed, wax-embedded tissue samples. This is the first time that this technique has been used for the detection of fish bacteria from diagnostic samples.     

Cadmium detoxification effect of Chinese parsley Coriandrum sativum in liver and kidney of rainbow trout Oncorhynchus mykiss

Although fish feed supplemented with Chinese parsley (CP), Coriandrum sativum, does not affect the health of rainbow trout Oncorhynchus mykiss, it has the remarkable effect of reducing the uptake of Cd (12–17% compared with controls without CP). Here, we measured the change in the amount of metallothionein (MT) in the liver and kidney using gel permeation chromatography-high-performance liquid chromatography (TSK GEL SW 3000 column). The increase in MT content over time in the Cd-fed groups corresponded to the change in Cd accumulation. Gel permeation was performed with soluble fractions from the fish liver and kidney, and changes in the Cd, Cu, and Zn concentrations over time for each fraction, and the relationship between the form of Cd that accumulated in tissues and its toxicity were measured. A nontoxic MT fraction of 7,000 Da and a high-molecular-weight fraction of approximately 60,000 Da were isolated from the soluble fractions. The induction of nontoxic MT–Cd was higher in the CP-supplemented groups than in the control groups. The synergetic actions of several compounds in CP may contribute to bind MT, thereby preventing accumulation of harmful heavy metals and essential metals in fish.