Arteriosclerosis in steelhead trout, Oncorhynchus mykiss (Walbaum): a developmental analysis

Arteriosclerosis in Oncorhynchur mykiss has been examined histologically in parr, smolts, spawning adults and post-spawning adults captured during downstream migration. All stages of fish from the Clearwater River in Idaho are characterized by the presence of lesions in the main vessel on the bulbus arteriosus. The incidence is lowest in parr, increases significantly in smolts, and is present in 100% of spawning fish. Regression of lesions was not observed in either hatchery or wild fish after spawning.     

三个虹鳟养殖群体遗传结构的微卫星分析

利用20个微卫星标记对3个虹鳟(Oncorhynchus mykiss)养殖群体进行遗传结构分析。结果表明,(1)3个群体检测的平均等位基因数为3·6～4·1,平均观测杂合度为0·5224～0·6328,平均期望杂合度为0·4736～0·5522,平均多态信息含量为0·4354～0·5084,说明这几个群体多态性属于偏高水平,遗传多样性高。通过d值,确定了Hard-Weinberg平衡的偏离情况,发现AY039638、AY039646在3个群体中都表现为不平衡。对3个群体的遗传距离进行了估算,并进行聚类分析,本溪的两个群体先聚为一支,再与渤海群体相聚,显示出明显的地理特征。(2)本溪虹鳟群体在位点AF352770出现部分等位基因消失的现象;AF352754在本溪群体中表现为位点消失,可作为区分本溪群体和渤海群体的分子标记。(3)综合评价3个群体,渤海站虹鳟群体的遗传多样性最高,与前人研究结果一致。     

Song environment affects singing effort and vasotocin immunoreactivity in the forebrain of male Lincoln's sparrows

Male songbirds often establish territories and attract mates by singing, and some song features can reflect the singer's condition or quality. The quality of the song environment can change, so male songbirds should benefit from assessing the competitiveness of the song environment and appropriately adjusting their own singing behavior and the neural substrates by which song is controlled. In a wide range of taxa, social modulation of behavior is partly mediated by the arginine vasopressin or vasotocin (AVP/AVT) systems. To examine the modulation of singing behavior in response to the quality of the song environment, we compared the song output of laboratory-housed male Lincoln's sparrows (Melospiza lincolnii) exposed to 1 week of chronic playback of songs categorized as either high or low quality, based on song length, complexity, and trill performance. To explore the neural basis of any facultative shifts in behavior, we also quantified the subjects' AVT immunoreactivity (AVT-IR) in three forebrain regions that regulate sociosexual behavior: the medial bed nucleus of the stria terminalis (BSTm), the lateral septum (LS), and the preoptic area. We found that high-quality songs increased singing effort and reduced AVT-IR in the BSTm and LS, relative to low-quality songs. The effect of the quality of the song environment on both singing effort and forebrain AVT-IR raises the hypothesis that AVT within these brain regions plays a role in the modulation of behavior in response to competition that individual males may assess from the prevailing song environment.     

Experimental studies on the effect of Gyrodactylus colemanensis and G. salmonis (Monogenea) on density of mucous cells in the epidermis of fry of Oncorhynchus mykiss

Fry of Oncorliynclius mykiss were experimentally infected with the ectoparasites Gyroducrylus rolenirmensis and Gyroducrylus salnionis (Monogenea) and changes in mucous cell density in the epidermis covering the caudal fin were monitored during an ensuing 42 day epidemic. At 10° C, infections of G. colemanensis rose to a recorded peak of 90 to 1 15 worms on day 27 post-infection and then declined toward extinction. Intensity of G. salmonis remained low throughout the experiment. Infection produced no clinical signs of disease and did not influence fry growth or survival. However, infected fish did have a 50% reduction in the number of mucous cells in the epidermis of the fin. The changes were first detected on day 24 post-infection and became increasingly pronounced during the subsequent 1 day period when parasite numbers declined drastically. The study hypothesizes that parasite activity on the surface indirectly leads to reduction in mucous cells through disruption of cell dynamics within the epidermis.     

Is there an optimal haematocrit for rainbow trout,Oncorhynchm mykiss (Walbaum)? An interpretation of recent data based on blood viscosity measurements

The viscosity of blood from rainbow trout was measured following manipulation of haematocrit by bleeding, hypoxia. exercise, and anaesthesia. Blood viscosity when measured at high shear rate (225 s ¹) was proportional to haematocrit, but the dependence of viscosity on shear rate was far less for swollen erythrocytes from exercised and anaesthetized trout. Erythrocyte swelling was most marked in exercised and anaesthetized trout, and is a confounding factor when considering the effect of haematocrit on viscosity.
The viscosity of blood with variable haematocrit, but constant mean cell Hb concentration, indicated that the relative oxygen transport capacity in trout was optimal at a haematocrit of 30%. Data from this, and earlier studies show that haematocrit in trout is variable and labile, yet none of the haematocrit values following manipulations are less than 85% of optimal. Optimal haematocrit is however, significantly higher than measured values from either cannulated or acutely venesected resting trout.     

Lysozyme activity in rainbow trout, Oncorhynchus mykiss (Walbaum), stressed by handling, transport and water pollution

Blood lysozyme activity was determined in rainbow trout that were stressed by handling and transport, or suffered from acute ammonia poisoning. To optimize the lysoplate assay, the pH dependence of the lysozyme activity was studied. For serum and kidney lysozyme, the pH optimum was 5.5 to 6.0. Ventilation frequency and plasma glucoseconcentration of the fish were used as stress indicators. Less severely stressful situations, such as 30 min of handling, brought about either a stimulation or a suppression of the lysozyme activity. Strong stressors, such as transport lasting 2 h or acute water pollution, reduced the lysozyme level significantly. The decrease of lysozyme activity due to handling and transport persisted for 24 h, but the activity returned to normal within 2 weeks following confrontation with the stressor.     

Stress effects on AVT and CRF systems in two strains of rainbow trout (Oncorhynchus mykiss) divergent in stress responsiveness

The aim for this study was to examine whether the F4 generation of two strains of rainbow trout divergent in their plasma cortisol response to confinement stress (HR: high responder or LR: low responder) would also differ in stress-induced effects on forebrain concentrations of mRNA for corticotropin-releasing factor (CRF), arginine vasotocin (AVT), CRF receptor type 1 (CRF-R1), CRF receptor type 2 (CRF-R2) and AVT receptor (AVT-R). In addition, plasma cortisol concentrations, brainstem levels of monoamines and monoamine metabolites, and behaviour during confinement were monitored. The results confirm that HR and LR trout differ in their cortisol response to confinement and show that fish of these strains also differ in their behavioural response to confinement. The HR trout displayed significantly higher locomotor activity while in confinement than LR trout. Moreover, following 180 min of confinement HR fish showed significantly higher forebrain concentrations of CRF mRNA than LR fish. Also, when subjected to 30 min of confinement HR fish showed significantly lower CRF-R2 mRNA concentrations than LR fish, whereas there were no differences in CRF-R1, AVT or AVT-R mRNA expression between LR and HR fish either at 30 or 180 min of confinement. Differences in the expression of CRF and CRF-R2 mRNA may be related to the divergence in stress coping displayed by these rainbow trout strains.     

The host specificity of Gyrodactylus salaris Malmberg (Platyhelminthes, Monogenea): susceptibility of Oncovhynchus mykiss (Walbaum) under experimental conditions

An experimental epidemiological approach was chosen to study the survival and infection dynamics of Gyrodactylus salaris on juvenile rainbow trout, Oncorhynchus mykiss , in the laboratory. A marked heterogeneity in the host stock was apparent. The rainbow trout could be divided into three groups on the basis of parasite survival and infection pattern on individually isolated fish: (1) hosts receptive to initial parasite attachment, but unreceptive to parasite establishment and reproduction; (2) hosts moderately susceptible to parasite establishment and reproduction, but which, after a period of restricted parasite population growth, responded, recovered and eliminated the parasites; and (3) hosts very susceptible to parasite infection and reproduction, but which, after a period of significant parasite population growth, responded, recovered and eliminated the parasites. These different patterns are considered to reflect genetic differences between host individuals. Parasite aggregation was also shown to be an important factor in the outcome of the host-parasite association. The parasites were finally eliminated on the individually isolated hosts, but not on hosts maintained in batches and so host population size and immigration of fresh. previously unexposed, hosts appeared to be important for growth and maintenance of the parasite population. The parasite was not found to cause host mortality. Rainbow trout was a suitable host for G. salaris , capable of transmitting the parasite to new localities as a consequence of stocking programmes or migratory behaviour.     

Acoustic measurement of trout distributions in Spada Lake, Washington, using stationary transducers

Diel patterns in the spatial distribution of rainbow and cutthroat trout were studied with stationary acoustic transducers, gillnets, and setlines in Spada Lake during summer thermal stratification. Four diel periods (dawn, 03.00–07.00 hours; day, 07.00–19.00 hours; dusk, 19.00–23.00 hours; night, 23.00–03.00 hours) and two horizontal strata (nearshore, bottom depth < 8 m; offshore, bottom depth > 8 m) were identified from fish distribution patterns. During the day, trout were almost exclusively offshore and their densities were highest at intermediate depths of the water column (4–16 m). From dusk to dawn, trout were in intermediate and shallow depths (0–4 m) of the offshore and nearshore strata. The mean depth of capture was significantly deeper for cutthroat trout (9 m) than for rainbow trout (3m). Both species primarily inhabited the epilimnion and metalimnion where temperature ranged from 22.5 to 11.0°C and were nearly absent from deeper, cooler waters. Dissolved oxygen levels ranging from 3.9 to 8.6 mg/l did not seem to influence trout distributions.     

Uptake of vitellogenin into oocytes during early vitellogenic development in the rainbow trout, Oncorhynchus mykiss (Walbaum)

Uptake of ³H-vitellogenin (³H-VTG) into oocytes of various sizes was investigated during early vitellogenic development in the rainbow trout, Oncorhynchus mykiss (Walbaum). Females were injected with ³H-VTG and uptake into oocytes of different sizes (<0.4,0.4–0.59, 0.6–0.79, 0.8–0.99 and 1.0 1.2 mm in diameter) measured. Oocytes measuring less than 0.6 mm in diameter appeared unable to sequester VTG and were therefore considered pre-vitellogenic. Oocytes measuring 0.6 mm or more all sequestered VTG. The larger the oocyte, the more ³H-VTG it sequestered, even when uptake was expressed per unit surface area. The latter observation could be due to an increase in the number of VTG receptors per unit surface area, an increase in the rate of turnover of the VTG receptor, greater access of VTG to the receptors as oocytes grow, or a combination of any of these factors. The data suggest that the ability to sequester VTG is developmentally regulated.     

气泡幕对虹鳟阻拦效果研究

为探究不同工况下气泡幕对虹鳟(Oncorhynchus mykiss)的阻拦效果, 在光照环境下分别采用4种孔距(1.0、2.0、3.0 和4.0 cm)、4种孔径(0.5、1.0、1.5 和2.0 mm)和3种气量(60、120 和180 L/min)产生的气泡幕, 用单一变量法对虹鳟进行阻拦实验, 获得所用工况中阻拦效果最优的工况; 在此基础上, 进行黑暗环境下气泡幕对虹鳟的阻拦效果实验。结果显示: (1)在光照环境下, 所用工况中阻拦效果最优的工况为孔径1.0 mm、孔距2.0 cm、气量120 L/min的气泡幕, 阻拦率为(96.32±3.99)%; (2)在黑暗环境下, 空白实验中游过气泡幕管道上方的鱼尾次数为光照环境下的63.37%, 阻拦率为(87.48±2.55)%。研究结果可为大水体养殖虹鳟的捕捞技术研究提供参考。     

Six diagnostic single nucleotide polymorphism markers for detecting introgression between cutthroat and rainbow trouts

Ten primer pairs were screened to develop single nucleotide polymorphism (SNP) TaqMan assays that will distinguish California golden trout and some rainbow trouts (Oncorhynchus mykiss sspp., O. m. aguabonita) from the Paiute and Lahontan cutthroat trouts (Oncorhynchus clarkii seleniris, O. c. henshawi). From these 10 primer pairs, one mitochondrial and five nuclear fixed SNP differences were discovered and developed into TaqMan assays. These six assays will be useful for characterizing and monitoring hybridization between these groups. Additional Oncorhynchus clarkii sspp. and Oncorhynchus mykiss sspp. were assayed to determine if these assays are useful in closely related species.     

Involvement of arginine vasotocin in reproductive events in the male newt Cynops pyrrhogaster

Effects of arginine vasotocin (AVT) on reproductive events such as courtship behavior, pheromone release, and spermatophore discharge were investigated in the male newt Cynops pyrrhogaster. AVT enhanced the incidence and frequency of androgen-induced courtship behavior. In this case, AVT was likely to act centrally because the behavior was evoked with a much smaller amount of AVT when the hormone was administered intracerebroventricularly than when given intraperitoneally. Involvement of endogenous AVT in spontaneously occurring courtship behavior was also evidenced by the fact that administration of a V1 (vasopressor) receptor antagonist, [d(CH2)5(1), Tyr(Me)2, Arg8-vasopressin] suppressed the expression of the courtship behavior. The water in which AVT-treated males had been kept showed considerable female-attracting activity as compared with the water in which saline-injected males had been kept. Moreover, the content of sodefrin, a female-attracting pheromone in the abdominal gland, was decreased by the intraperitoneal injection of AVT, suggesting that the neurohypophyseal hormone stimulated the release of sodefrin from the abdominal gland into the water. AVT induced contraction of the excised abdominal gland concentration-dependently, and, again, the V1 receptor antagonist suppressed the AVT-induced contraction. Thus, we concluded that AVT induces the pheromone discharge, acting peripherally on a contractile structure of the abdominal gland. AVT was also found to induce spermatophore deposition in the male kept in the absence of the female. Administration of the V1 receptor blocker to the sexually developed males suppressed the spermatophore deposition. All these results indicate the involvement of AVT in reproductive events acting centrally and peripherally.     

虹鳟补体活化调节因子CD46的分子特征及功能

为进一步研究硬骨鱼类中补体活化调节因子的分子特征和功能, 研究克隆了虹鳟(Oncorhynchus mykiss)的CD46基因, 对其分子特征进行了系统分析, 结果显示: 虹鳟CD46基因由10个外显子和9个内含子组成, cDNA序列全长2812 bp, 编码317个氨基酸, 蛋白序列由1个信号肽、4个SCR结构域、1个跨膜区和1个胞内区组成, 预测分子量为33.9 kD。基因组共线性分析显示, 虹鳟CD46基因位于16号染色体, 其基因座在脊椎动物中具有保守的共线性。组织和白细胞亚群表达分析显示, 虹鳟CD46基因在各种组织和白细胞亚群中均有表达。为了进一步阐明虹鳟CD46的免疫功能, 研究原核表达纯化了标签蛋白GST和融合蛋白GST-CD46。溶血活性实验表明, 与GST相比, GST-CD46能够显著抑制虹鳟血清对兔红细胞的溶血活性, 且呈现剂量依赖效应, 表明虹鳟CD46是补体活化的调节因子。此外, 研究用HEK293T细胞过表达了GFP和GFP-CD46。细胞损伤实验显示, 与GFP相比, GFP-CD46能够显著抑制虹鳟血清对HEK293T细胞的损伤, 进一步表明虹鳟CD46是补体活化的调节因子, 能够保护细胞免受补体系统的损伤。总之, 研究不仅增加了对虹鳟CD46的分子特征和功能的认知, 还为深入研究此分子的免疫功能及其在抗病免疫中的应用提供了理论基础。     

基于UPLC-Q-TOF/MS代谢组学探究虹鳟鳃对热应激的生理反应

全球气温日趋升高导致的水温上升可引起虹鳟(Oncorhynchus mykiss)代谢紊乱, 为解析其在热应激下的代谢变化特征, 研究基于UPLC-Q-TOF/MS代谢组学技术, 探究了虹鳟鳃靶器官在高温暴露(20℃和24℃)及恢复到初始温度(14℃)下的代谢生理反应。研究结果表明, 与对照组相比, 在20℃、24℃高温组和14℃恢复组中分别鉴定出128、130和108种差异显著代谢物。在高温暴露下, 亚油酸、花生四烯酸等与脂质代谢相关的代谢物及还原型辅酶Ⅱ(NADPH)、谷胱甘肽(GSH)、谷胱甘肽二硫化物(GSSG)等与细胞氧化还原状态相关的代谢物均发生显著改变。富集分析表明这些代谢物主要涉及虹鳟鳃的甘油磷脂代谢、鞘脂代谢、亚油酸代谢、花生四烯酸代谢、磷酸戊糖途径和谷胱甘肽代谢等代谢通路, 但在温度恢复到14℃后, 除鞘脂代谢外, 其他代谢途径均未恢复至正常状态。上述结果表明, 热激导致了虹鳟鳃靶器官的脂质代谢紊乱, 可能导致鳃细胞膜的结构和功能的损伤, 诱使鳃细胞出现炎症反应, 并产生免疫应答。同时, 虹鳟鳃细胞通过磷酸戊糖途径产生的NADPH来调节谷胱甘肽代谢中GSH/GSSG比值以提高细胞的抗氧化能力来对抗氧化应激, 防止细胞凋亡。研究结果为理解热应激下虹鳟的生理反应提供了科学依据。     

The distribution of corticotropin-releasing factor-immunoreactive neurons and nerve fibers in the brain of the snake,Natrix maura

Summary The anatomical distribution of neurons and nerve fibers containing corticotropin-releasing factor (CRF) has been studied in the brain of the snake, Natrix maura, by means of immunocytochemistry using an antiserum against rat CRF. To test the possible coexistence of CRF with the neurohypophysial peptides arginine vasotocin (AVT) and mesotocin (MST) adjacent sections were stained with antisera against the two latter peptides. CRF-immunoreactive (CRF-IR) neurons exist in the paraventricular nucleus (PVN). In some neurons of the PVN, coexistence of CRF with MST or of CRF with AVT has been shown. Numerous CRF-IR fibers run along the hypothalamo-hypophysial tract and end in the outer layer of the median eminence. In addition, some fibers reach the neural lobe of the hypophysis. CRF-IR perikarya have also been identified in the following locations: dorsal cortex, nucleus accumbens, amygdala, subfornical organ, lamina terminalis, nucleus of the paraventricular organ, nucleus of the oculomotor nerve, nucleus of the trigeminal nerve, and reticular formation. In addition to all these locations CRF-IR fibers were also observed in the lateral septum, supraoptic nucleus, habenula, lateral forebrain bundle, paraventricular organ, hypothalamic ventromedial nucleus, raphe and interpeduncular nuclei.     

Construction and characterization of BAC libraries for three fish species; rainbow trout, carp and tilapia

Bacterial artificial chromosome (BAC) libraries are important tools for genomic research. We have constructed seven genomic BAC libraries from three fish species, rainbow trout (Oncorhynchus mykiss), carp (Cyprinus carpio) and tilapia (Oreochromis niloticus). The two rainbow trout BAC libraries have average insert sizes of 58 and 110 kb. The average size of inserts in the carp BAC library is 160 kb. The average insert sizes of the four tilapia BAC libraries are 65, 105, 145 and 194 kb, respectively. These libraries represent good coverage of each genome (2-64 x coverage). The libraries can be screened by conventional colony hybridization and provide a starting point for the construction of high-density filtres or polymerase chain reaction (PCR) screening approaches. These BAC libraries will facilitate the positional cloning of quantitative trait loci (QTLs) for a variety of economically important traits in these species.