重金属离子Cd2+对泥蚶鳃及肝脏细胞显微和超微结构的影响

研究了镉(Cd)的各个不同暴毒浓度(5,15,45,90μg/L)对泥蚶鳃和肝脏组织显微结构的影响,及高浓度暴毒情况下对泥蚶鳃和肝脏组织超微结构的影响。实验处理时间为96 h。结果发现,随着暴毒浓度的升高,鳃丝内腔隙肿胀,内有血细胞堆积,最后鳃丝出现断裂融合,其上的呼吸上皮细胞脱落,超微结构发现鳃上皮细胞中出现黑色嗜锇性物质,且次级溶酶体和线粒体数量都增加,最后细胞内出现空泡现象;Cd对肝脏显微结构的影响不大,仅出现了黄色沉积物,超微结构表明,肝细胞内亦出现了嗜锇性物质,次级溶酶体亦大量增加,这些现象与鳃丝内出现的情况一致,且肝细胞核还出现变形皱缩,甚至产生空泡,细胞出现了不可逆的损伤。最为敏感的线粒体和内质网没有出现损伤现象,研究推测可能是由于机体内Cd诱导产生的抗氧化活性酶作用的结果。     

鲮鱼肝细胞超微结构研究

应用透射电镜对鲮鱼肝脏细胞进行了观察。结果表明, 正常状态下肝细胞具有典型的单核现象, 核仁、核膜、核孔等都清晰可见, 肝细胞含有线粒体、粗面内质网、溶酶体、糖原等细胞器和内容物, 胆小管主要为胞内胆小管。饥饿4周后, 鲮鱼肝细胞所含的细胞器总数目减少, 核周粗面内质网含量降低, 线粒体出现多种形状, 其间电子密度较低, 嵴更加稀疏, 溶解, 无全嵴, 其最大变化是肝细胞内的糖原几乎消失, 出现大量的溶酶体。再恢复投喂4周后, 鲮鱼肝细胞各种细胞器均恢复到正常水平, 鲮鱼的糖原大量出现, 占据细胞大量空间。表明鲮鱼肝细胞代谢与细胞器数目和内容物密切相关, 在一段时间的饥饿胁迫之后, 其肝脏能过量地贮存糖原作为能源物质。     

鲮鱼肝细胞超微结构研究

应用透射电镜对鲮鱼肝脏细胞进行了观察.结果表明,正常状态下肝细胞具有典型的单核现象,核仁、核膜、核孔等都清晰可见,肝细胞含有线粒体、粗面内质网、溶酶体、糖原等细胞器和内容物,胆小管主要为胞内胆小管.饥饿4周后,鲮鱼肝细胞所含的细胞器总数目减少,核周粗面内质网含量降低,线粒体出现多种形状,其间电子密度较低,嵴更加稀疏,溶解,无全嵴,其最大变化是肝细胞内的糖原几乎消失,出现大量的溶酶体.再恢复投喂4周后,鲮鱼肝细胞各种细胞器均恢复到正常水平,鲮鱼的糖原大量出现,占据细胞大量空间.表明鲮鱼肝细胞代谢与细胞器数目和内容物密切相关,在一段时间的饥饿胁迫之后,其肝脏能过量地贮存糖原作为能源物质.     

漂白粉蓄积对日本蟳4种组织超微结构的影响

根据急性毒性试验确定了漂白粉对日本蟳的安全质量浓度为0．6mg／L，采用透射电镜技术研究了该质量浓度下漂白粉蓄积对日本蟳4种组织细胞(肌肉、鳃、肝胰脏和心脏)超微结构的影响。试验结果表明：漂白粉主要损伤日本蟳的鳃和肝胰脏，其次是心脏，肌肉基本无损害。鳃细胞的损害表现为鳃丝水肿，细胞器溶解，角质层变薄，线粒体、内质网的肿胀、解体；肝胰脏细胞的主要损害特征为肝管微绒毛减少、线粒体水肿解体、内质网扩张、细胞核空泡化、核膜水肿、脂肪滴增加；心脏细胞的毒理变化为线粒体内嵴肿胀、瓦解，肌原纤维不规则，内质网溶解。安全质量浓度的漂白粉经16d蓄积后，虽没有导致日本蟳死亡，但已对其体内的鳃、肝胰脏、心脏等组织细胞的超微结构产生了不同程度的影响。     

达氟沙星对施氏鲟的急性毒性及组织残留检测

将施氏鲟(Acipenser scherenscki,体重75～95g)暂养于室内玻璃水族箱内7d后进行实验。所用达氟沙星(Danofloxacin)纯度为99．5％。分别以剂量800、l040、1352、1757．60、2284．88和2970．34mg／(kg体重)对施氏鲟进行口灌,以400、440、484、532．4、585．6和644．2mg,／(kg体重)进行腹腔注射达氟沙星水溶液,给药后连续10d观察实验鱼的行为及死亡情况。用改进的寇氏法计算得出施氏鲟腹腔注射达氟沙星的LD50为429．47mg／(kg体重),LD50的95％可信限为425．22～433．66mg／(kg体重)。口灌达氟沙星LD50为l502．10mg／(kg体重),95％可信区间范围为1307．89～1738．60mg／(k体重)。对染毒死亡鱼及对照组实验鱼分别进行组织切片,并分别进行光镜及电镜观察。光镜观察结果表明,染毒死亡鱼的肝细胞的索状结构消失,肝组织呈弥漫性坏死,肝细胞肿胀,有些肝细胞失去正常的多角形形状,细胞核肿胀变形;窦状隙腔变窄,其中的红细胞形状也不同程度发生了改变。染毒死亡鱼的肝细胞的超微结构显示,肝细胞内充满脂滴,肝细胞核萎缩、消失,线粒体脊断裂,线粒体破裂,粗面内质网结构疏松有断裂,滑面内质网数量明显减少,溶酶体破裂。用高效液相色谱法测定以10mg/(k体重)剂量连续4次口灌给药后,达氟沙星在施氏鲟血浆、肝脏、肾脏、肌肉和鳃组织中的药物水平,实验结果表明,达氟沙星在施氏鲟全身组织均有分布,用药第9天后达氟沙星在施氏鲟体内消除速度很快,各组织药物水平均在最低检测限以下,其中肌肉和鳃在第6天就可降到0．005μg/(g组织)以下,肝脏和肾脏在第9天、血浆在第7天均检测不到,检测发现肝脏、肾脏中的药物水平远远高于血浆、肌肉和鳃组织中的血药水平。建议肝脏和肾脏可作为达氟沙星的残留检测的指示组织。     

硫酸铜蓄积对日本[虫寻]4种组织细胞超微结构的影响观察

通过蓄积毒性试验，研究了安全质量浓度（0．5mg·L^-1）下硫酸铜（CuSO4·5H2O）对日本[虫寻]（Charybdis japonica）4种组织细胞（肌肉、鳃、肝胰脏和心脏）超微结构的影响。结果表明，铜离子（Cu^2＋）蓄积主要损伤的组织是鳃和肝胰脏，其次是心脏和肌肉。鳃细胞的损害表现为鳃丝水肿，细胞器溶解，角质层变薄，线粒体、内质网的肿胀、解体；肝胰脏细胞的主要损害特征为肝管微绒毛减少，线粒体水肿解体，内质网扩张，细胞核空泡化，核膜水肿和脂肪滴增加；心脏细胞的毒理变化为线粒体内嵴肿胀、瓦解，肌原纤维不规则，内质网溶解。0．5mg·L^-1的CuSO4·5H2O经过16d蓄积后虽然没有导致日本[虫寻]死亡，但已经对其体内组织细胞的超微结构产生了不同程度的影响。     

温州养殖文蛤死亡原因调查及组织超微病理分析

针对近年来温州市龙湾区养殖文蛤发生死亡的特点进行了病原调查和组织病理分析。通过定期采集样品,运用显微镜检、超微结构分析等方法,探讨造成文蛤死亡的原因。调查发现龙湾区文蛤病害爆发主要集中在每年7月份,具有明显的季节性和流行性。患病文蛤无寄生虫及病毒的存在,超微病理观察发现,肝胰腺、肌肉、鳃等组织均出现了细胞核染色质边聚,内质网肿胀,排列稀疏无序,线粒体大量凋裂,形成较多空泡等现象。分析表明,引起文蛤死亡的原因主要是环境的变化带来的应激性病原体侵入有关,同时与文蛤本身品质,养殖密度,养殖管理等因素密切相关。     

镉对弹涂鱼肝细胞超微结构影响的初步观察

研究了镉（Cd）的两种离子浓度（1mg·L^-1、0．1mg·L^-1）对弹涂鱼肝细胞超微结构的影响。采用体外暴露方法，将实验用弹涂鱼暴露于lmg·L^-1Cd^2＋、0．1mg·L^-1Cd^2＋中，暴露10d后取肝脏制备电镜样品，应用透射电镜观察肝细胞超微结构。结果表明，与镉的对照组相比，在0．1mg·L^-1Cd^2＋浓度暴露条件下，弹涂鱼肝脏细胞内的细胞器受到不同程度的损伤，线粒体扭曲变形，内质网膨胀，细胞核变形，核膜肿胀。在1mg·L^-1Cd^2＋浓度组，几乎所有的细胞器都受到严重影响，细胞结构遭到严重破坏。肝细胞细胞质内容物大量泄露，胞浆空泡化，细胞器极少，残存细胞器的结构不完整。研究表明随着Cd^2＋暴露浓度的增高，肝细胞超微结构的受损程度逐渐加深，损伤不可逆转。分析认定Cd^2＋对肝细胞的损伤是由脂质过氧化造成。     

漂白粉蓄积对日本昭示(虫寻)4种组织超微结构的影响

根据急性毒性试验确定了漂白粉对日本的安全质量浓度为0.6mg/L,采用透射电镜技术研究了该质量浓度下漂白粉蓄积对日本4种组织细胞(肌肉、鳃、肝胰脏和心脏)超微结构的影响。试验结果表明:漂白粉主要损伤日本的鳃和肝胰脏,其次是心脏,肌肉基本无损害。鳃细胞的损害表现为鳃丝水肿,细胞器溶解,角质层变薄,线粒体、内质网的肿胀、解体;肝胰脏细胞的主要损害特征为肝管微绒毛减少、线粒体水肿解体、内质网扩张、细胞核空泡化、核膜水肿、脂肪滴增加;心脏细胞的毒理变化为线粒体内嵴肿胀、瓦解,肌原纤维不规则,内质网溶解。安全质量浓度的漂白粉经16d蓄积后,虽没有导致日本死亡,但已对其体内的鳃、肝胰脏、心脏等组织细胞的超微结构产生了不同程度的影响。     

硫酸铜蓄积对日本4种组织细胞超微结构的影响观察

通过蓄积毒性试验,研究了安全质量浓度(0.5mg.L-1)下硫酸铜(CuSO4.5H2O)对日本(Charybdisjaponica)4种组织细胞(肌肉、鳃、肝胰脏和心脏)超微结构的影响。结果表明,铜离子(Cu2+)蓄积主要损伤的组织是鳃和肝胰脏,其次是心脏和肌肉。鳃细胞的损害表现为鳃丝水肿,细胞器溶解,角质层变薄,线粒体、内质网的肿胀、解体;肝胰脏细胞的主要损害特征为肝管微绒毛减少,线粒体水肿解体,内质网扩张,细胞核空泡化,核膜水肿和脂肪滴增加;心脏细胞的毒理变化为线粒体内嵴肿胀、瓦解,肌原纤维不规则,内质网溶解。0.5mg.L-1的CuSO4.5H2O经过16d蓄积后虽然没有导致日本死亡,但已经对其体内组织细胞的超微结构产生了不同程度的影响。     

miR‐122‐5p as a plasma biomarker of liver injury in fish exposed to microcystin‐LR

Recent studies have shown the presence of large amounts of microRNAs (miRNAs; miRs) from damaged cells in the peripheral blood. In this study, we investigated the levels of miRNAs circulating in the blood plasma of whitefish (Coregonus lavaretus) after exposure to microcystin‐LR. We used real‐time PCR to examine the relative expression of plasma levels of 4 miRNAs (miR‐122‐5p and let‐7c‐5p, the liver‐enriched microRNAs, miR‐148a‐3p which promotes the hapatospecific phenotype in mammals, and miR‐92a‐3p, a cell proliferation and angiogenesis promoter, potentially hepatocarcinogenic) during the first 48 h after exposure to MC‐LR. We observed a rapid increase of miR‐122‐5p levels 8 h after exposure (P < 0.05), which continued to the end of the experiment. Our results demonstrated that the plasma miR‐122‐5p was indicative of MC‐LR‐induced liver injury, exhibiting areas under the curve close to 1 in ROC analysis (AUC = 0.976, P < 0.001). Although plasma levels of miR‐148a‐3p and miR‐92a‐3p were significantly elevated by the end of the experiment, their discriminative power was lower than reported for the miR‐122‐5p. Based on these results and reports on miRNA‐based diagnosis of liver injuries in mammals, plasma miR‐122‐5p could be considered as a robust, new generation diagnostic biomarker in fish, helpful for the non‐invasive diagnosis of liver damage.     

Potent activation of tumor necrosis factor-α production by a polysaccharide found in a marine brown alga Chordaria flagelliformis

In a search for new antitumor drugs, we collected 334 species of marine algae from the Japanese coastline and screened these for tumor necrosis factor-alpha (TNF-α) production-promoting activity in an in vitro assay with the mouse macrophage-derived cell line RAW264.7. A phosphate-buffered saline extract from the marine brown alga Chordaria flagelliformis was found to strongly promote TNF-α activity. Further purification of this alga by precipitation with ethanol and two-step chromatography yielded an approximately 3,000-kDa polysaccharide as the active substance, which we designated MC25. MC25 exhibited TNF-α production-promoting activity in RAW264.7 cells in a dose-dependent manner, and we established its temperature and pH stabilities and non-cytotoxic character. Its molecular structure is similar to that of a fucoidan in terms of composition, including glucose, fucose, xylose, and uronic acid. The molecule hydrolyzed to a size of 180–370 kDa still retained strong TNF-α production-promoting activity, suggesting that MC25 might be a promising candidate in the development of novel antitumor agents.     

Aflatoxins from Moldy Corn Cause No Reductions in Channel Catfish Ictalurus punctatus Performance

Aflatoxins are a group of mycotoxins produced by the mold organisms Aspergillus flavus and A. parasiticus on feed grains and oil seeds such as corn, peanuts, and cottonseed. Research conducted in aquaria, about 15 yr ago, demonstrated that channel catfish Ictalurus punctatus are very tolerant to dietary aflatoxin B₁ (AFB₁) from a purified source. To evaluate the effect of feeding diets containing aflatoxin from a natural source, moldy corn (MC) naturally contaminated with a high concentration (550 pg/kg) of total aflatoxins was incorporated into practical diets. The diets were fed to Juvenile catfish in two experiments. Experiment 1 consisted of feeding catfish (mean body weight 7.1 g/fish) four diets containing 20% or 40% of two lots of corn; one with no apparent mold contamination, which was designated as clean corn (CC), or the previously described MC. Each diet was fed twice daily to five 100-L aquaria of 20 fish each for 12 wk. Experiment 2 consisted of three diets containing either 50% CC or MC, or a combination of 25% CC and 25% MC prepared by the cooker-extrusion method. Each diet was fed once daily for 130 d to five replicate 0.04-ha ponds of catfish fingerlings. Results of these experiments indicate that feeding diets containing aflatoxin from moldy corn does not affect channel catfish weight gain, feed consumption, feed efficiency, survival, hematocrit, or hepatosomatic ratio. No liver abnormalities were observed upon gross examination. Levels of aflatoxin were reduced approximately 63% in the diets used in experiment 2 after exposure to the high temperature (ca. 120 C) of the cooker-extrusion process used to manufacture commercial catfish diets.     

太湖微囊藻毒素在罗非鱼体内的累积及生物降解的初步研究

为了解蓝藻水华期间微囊藻毒素在罗非鱼体内的分布及累积传递过程，2008年6月至8月采集了高密度蓝藻池塘及太湖网箱内的鱼样及水样，用ELISA法对鱼样和水样进行微囊藻毒素MC-LR含量的检测。结果表明：池塘水体微囊藻毒素MC-LR含量变化范围在0.123～0.514ug/L间，MC-LR含量随着藻密度的下降而降低，对照组水体MC-LR浓度显著高于实验组MC-LR含量。池塘鱼体肌肉组织微囊藻毒素MC-LR累积含量在1.194～3.615ng/g间，肝脏组织微囊藻毒素MC-LR累积含量显著高于肌肉组织。将池塘与网箱罗非鱼转至无微囊藻水体中暂养，跟踪检测MC-LR含量变化，池塘和网箱鱼体肌肉组织微囊藻毒素MC-LR含量均低于人体每日可耐受摄入量，而肝脏组织藻毒素MC-LR含量则分别需要经过10～20天自然生物降解后降低至安全摄入量之下。并讨论了微囊藻毒素在鱼体内的组织分布与食物链中的累积传递。     

微囊藻毒素急性暴露对斑马鱼卵巢的损伤效应

为了探究蓝藻(Cyanobacteria)大量爆发产生的微囊藻毒素(microcystins,MCs)对鱼类的生殖毒性,以斑马鱼(Danio rerio)为实验对象,采取腹腔注射毒性最强的microcystin-LR(MC-LR)方式,研究MC-LR对斑马鱼卵巢的损伤效应及其作用机制。对性成熟雌性斑马鱼腹腔注射50μg/kg和200μg/kg MC-LR,在注射3、9、24、48 h后取卵巢分析其生理活性指标的变化。结果显示,染毒24 h后,200μg/kg剂量组斑马鱼性腺指数(gonad somatic index,GSI)14.14与对照组性腺指数16.98相比显著降低(P<0.05),其他组别无显著变化;卵巢发生卵母细胞空泡化、卵母细胞膜与滤泡细胞层连接组织缺失等病理现象;MC-LR显著抑制了斑马鱼卵巢蛋白磷酸酶(protein phosphatase 2A,PP2A)活性,并激活促成熟因子(maturation promoting factor,MPF)活性;MC-LR处理后,斑马鱼卵巢内丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)家族中的p 38MAPK、ERK1/2的转录水平显著上调,JNK未发生显著变化。研究表明,MC-LR抑制斑马鱼卵巢PP2A活性,并激活MPF活性与MAPK信号通路中ERK1/2与p 38MAPK的转录水平,进而干扰其卵母细胞的发育进程并产生生殖毒性。     

Physiological studies on the effect of copper nicotinate (Cu–N complex) on the fish, <Emphasis Type="Italic">Clarias gariepinus</Emphasis>, exposed to mercuric chloride

Female catfish, Clarias gariepinus, were collected from the Nile River at Assiut region, were divided into 7 groups. The first group was left as control, and the second was treated with mercuric chloride (MC) for 3 weeks following by normal water for 1 week. The third, fourth and fifth groups were provided by MC (150 μg/ l of water). This treatment was continued for 3 weeks. Then, the fish were received CN instead of MC, for 1 week, with 15 and 25 mg CN/100 g wet food. The fifth fish group received diet supplemented with vit E (α-tocopherol) (100 mg/kg wet diet), for 1 week, instead of MC treatment. Vitamin E was used as standard antioxidant drug. Following 3 weeks of normal ambient water, the sixth and seventh aquaria received only CN for 1 week, with 15 and 25 mg CN respectively/100 g wet food, respectively. At the end of the experiment, Samples of liver, kidneys (posterior part), gills (right gills) and ovary were excised. The measurement included the oxidative stress parameters: carbonyl protein and total peroxide and the antioxidant enzyme activities superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) in all selected organs. MC treatment induced harmful effect in fish, probably due to its enhancing effect on reactive oxygen species (ROS) production in fish organs especially the respiratory and osmoregulatory organs namely gills. The result suggests that this gill damage may exert hypoxic case, anoxia for different organs and some Cu excretion resulting in a magnification of ROS overproduction. Also, the observed oxidative stress in ovary tissue of MC-treated fish may affect fish fertility. The addition of CN in fish diets could protect the fish C. gariepinus against MC-induced oxidative damage showing recovery of fish organs. It could suggest that the detoxifying mechanism of action of CN is mainly due to its scavenging activity of free radicals rather than tissue healing.     

怀头鲇、鲇及其杂交F↓（1）肝、胰脏胚后发育及卵黄吸收方式

利用形态学和组织连续切片技术,对怀头鲇（Silurus soldatovi）、鲇（Silurus asotus）及其杂交F1的肝、胰脏胚后发育和卵黄吸收方式进行对比观察.结果表明,3种鲇出膜后约2天在心脏后方有一肝细胞团,3天后肝细胞团逐步增大,4天后肝分叶.以后随着各种鲇生长速度不同肝、胰脏发育程度也不同.3种鲇的胰脏均为紧凑型,卵黄囊依照先卵黄球、后脂肪的顺序被吸收,3种鲇只有怀头鲇和杂交F1代卵黄吸收方式相同.出膜后4天,各鲇的卵黄均被全部吸收,腹腔上部大部分空间为肝脏占有,同时腹腔内出现结构简单的胃及肠.研究还发现肝脏的发育与卵黄囊密切有关.[中国水产科学,2006,13（3）：460-465]     

Effects of feeding a high level of D-glucose on liver function in juvenile white sturgeon (Acipenser transmontanus)

Juvenile white sturgeon (Acipenser transmontanus) were fed three isonitrogenous and isoenergetic diets containing either 35% D-glucose (HC), a mixture of 20% dextrin and 10% cellulose (MC), or 23% cellulose (LC), to investigate the effects of dietary carbohydrate on liver function. After 8-week feeding, body weight gain of fish fed the HC diet was consistently higher than that of fish fed the MC and LC diets, but was not significantly different from the MC-fed fish. Fish fed the HC diet had significantly (p < 0.05) higher feed efficiencies and liver glycogen concentrations than fish fed the MC and LC diets. Sturgeon were injected intravenously with 10 mg kg^-1 body weight of sulfobromophthalein (BSP) and post-injection blood taken from the caudal vein at 15, 30, 60, and 120 min. No significant differences in plasma BSP concentrations were found among the treatments at these times. Plasma hemoglobin and activities of aspartate and alanine aminotransferase were not affected by the diets. This study suggests that the HC diet does not adversely affect liver function or weight gain. Inclusion of high dietary levels of digestible and inexpensive carbohydrates in commercial sturgeon feeds seems promising, but long-term feeding trials should be conducted to confirm this assertion.     

Evidence of rapid transfer and bioaccumulation of Microcystin‐LR poses potential risk to freshwater prawn Macrobrachium rosenbergii (de Man)

Microcystins accumulate in aquatic organisms and can be transferred to higher trophic levels, eventually affecting vector animals and consumers. We examined three levels of an aquatic food chain (Microcystis aeruginosa, Daphnia magna and Macrobrachium rosenbergii) to identify the transfer efficiency and risk of microcystin on prawns. Samples were analysed using ultra performance liquid chromatography‐mass spectrometry (MS)/MS and microcystin‐LR (MC‐LR) distributions in prawn tissues were studied. The results showed that prawns accumulate MC‐LR both directly from M. aeruginosa and indirectly through D. magna which was pre‐exposed to M. aeruginosa. MC‐LR was detected in the gills, digestive tracts and hepatopancreas of the prawns 2 h after exposure. MC‐LR accumulated in prawns to 0.49 ± 0.04 μg g^?1 dry weight in hepatopancreas within 24 h, while it was not detected in muscle samples, and rarely appeared in blood samples in such a short period. Although MC‐LR was not detected in muscle, the head including hepatopancreas of the prawns accumulated troublesome amounts of MC‐LR. These results demonstrate that microcystis blooms in prawn farming potentially pose a risk to human consumers, although prawns may be exposed to the bloom for a very short time, hence regular monitoring of blue green algae population is recommended.     

Infectious pancreatic necrosis virus in Atlantic salmon, Salmo salar L., post-smolts in the Shetland Isles, Scotland: virus identification, histopathology, immunohistochemistry and genetic comparison with Scottish mainland isolates

During mid-June 1999 peak mortalities of 11% of the total stock per week were seen at a sea cage site of Atlantic salmon, Salmo salar L., post-smolts in the Shetland Isles, Scotland. Virus was isolated on chinook salmon embryo (CHSE) cells in a standard diagnostic test and infectious pancreatic necrosis virus (IPNV) identified by enzyme-linked immunosorbent assay. IPNV was confirmed as serogroup A by a cell immunofluorescent antibody test using the cross-reactive monoclonal antibody AS-1. Four weeks after the main outbreak, virus titres in surviving moribund fish were assayed at >10(10) TCID50 g(-1) kidney. Histopathology of moribund fish was characterized by pancreatic acinar cell necrosis and a marked catarrhal enteritis of the intestinal mucosa. In the liver, necrosis, leucocytic infiltration and a generalized cell vacuolation were noted. IPNV-specific immunostaining was demonstrated in pancreas, liver, heart, gill and kidney tissue. The nucleotide sequence of the coding region of segment A was determined from the Shetland isolate. A 1180 bp fragment of the VP2 gene of this isolate was compared with a 1979 reference isolate from mainland Scottish Atlantic salmon, La/79 and another more recent mainland isolate, 432/00. Both A2 isolates were derived from carrier fish without signs of IPN and serotyped by a plaque neutralization test. The Shetland isolate shows a different nucleotide and amino acid sequence compared with the two isolates from carrier fish. These latter isolates showed identical amino acid sequences in the fragment examined, despite the 21 years separating the isolations. Sequence comparisons with other A2 (Sp) isolates on the database confirm all three Scottish isolates are A2 (Sp).