黄鳝隐藏新棘虫的血液与组织病理研究

采用扫描电镜、组织切片和血液检测方法对感染隐藏新棘虫(Neosentis celatus)的黄鳝(Monopterusalbus)肠道和血液进行病理研究,结果发现隐藏新棘虫的吻钩在肠道内壁形成大小不一的洞,肠绒毛脱落,组织纤维化;受损伤的肠道皱褶比较凌乱,柱状上皮细胞变性、坏死、脱落。损伤深入到固有层,使黏膜层与固有层分离,固有层被水肿液填充,有大量炎症细胞浸润;感染隐藏新棘虫的黄鳝肾脏和肝脏功能未受影响。     

微孢子虫感染三疣梭子蟹的肌组织病理及其免疫相关酶活性

利用纯化的三疣梭子蟹(Portunus trituberculatus)"牙膏病"病原微孢子虫对三疣梭子蟹进行人工感染实验,观察感染蟹肌肉组织的临床变化及其超微病理,并测定感染后肌肉和肝胰腺组织酸性磷酸酶(ACP)和超氧化物歧化酶(SOD)不同时间点的活性。结果发现,微孢子虫肌肉注射感染三疣梭子蟹后,蟹的肌纤维透明度随着感染时间的延长逐渐下降并向白浊化转变。超微病理显示,注射感染176 h后,肌细胞内存在大量微孢子虫,感染蟹肌细胞出现核膜肿胀和线粒体双层膜破裂等病变,严重者肌纤维发生断裂、溶解,电镜下可见大量高电子密度的溶解灶。免疫相关酶活分析显示,微孢子虫感染后,三疣梭子蟹肌肉和肝胰腺组织ACP和SOD活性均呈现两个显著上升过程(P0.05);肝胰腺组织ACP活性始终高于肌肉组织,而两种组织的SOD活性恰好相反;感染蟹肌肉组织内两种酶活性几乎始终大于正常对照组。酶活的变化可能与蟹微孢子虫的肌肉组织亲嗜性及其增殖速度较慢等特点有关。     

Effects of dietary starch levels on the growth,plasma metabolites and expression of genes involved in glucose transport and metabolism in Pelodiscus sinensis juveniles

A 10‐week feeding trial was conducted to evaluate the growth performance, glucose transport and metabolism of Chinese soft‐shelled turtles (Pelodiscus sinensis) exposure to graded levels of dietary starch (0.52%, 7.43%, 14.74%, 22.99% and 31.38%). The 360 turtles (initial body weight, 12.94 ± 0.50 g) with 12 replicates were randomly assigned to five experimental diets. The highest weight gain and specific growth rate (SGR) were observed in 7.43% group and the lowest in 31.38% group. The protein efficiency ratio, whole‐body lipid contents, hepatic glycogen contents and the 4‐hr postprandial plasma glucose levels were significantly increased with the increment of starch levels (p < .05). In contrast, the daily feed intake and feed conversion ration were significantly declined (p < .05). The mRNA levels of glucose transporter 2, glucokinase, pyruvate kinase, malic enzyme and acetyl‐CoA carboxylase alpha genes in the liver significantly increased as the increase in starch levels at 4‐hr and 24‐hr post feeding (p < .05). No significant differences were observed in the expression of gluconeogenesis genes at each time point (p > .05). These results suggested that dietary addition of starch up‐regulated hepatic glycolysis, glycogenesis and lipogenesis genes expression, but the deficient response of gluconeogenesis to dietary starch might be part of the causes limited the starch utilization. Based on the secondary polynomial regression of SGR, y = ?0.0011x² + 0.028x + 1.63 (R² = 0.9292), the 12.73% inclusion level of dietary starch was recommended in juvenile turtles.     

Fluctuation in the distribution of low-salinity water in the North Equatorial Current and its effect on the larval transport of the Japanese eel

Surface water in the North Equatorial Current (NEC) is composed of southern low‐salinity water diluted by precipitation to less than 34.2 psu and northern, high‐salinity tropical water greater than 34.8 psu. Analyses of 27‐year historical data, observed in winter and summer along the longitude 137°E by the Japan Meteorological Agency, shows that an obvious salinity front (34.5 psu) generated by the two water masses was usually located around 15°N. However, the salinity front has been moving northward during the past three decades. El Niño/Southern Oscillation (ENSO) affected salinity in the surface layer, while temperature changed in the middle layer. The salinity front sometimes moved southward, mainly south of 5°N, and the movement was well correlated with the southern oscillation index (SOI). Because precipitation at Yap (9.5°N, 138.1°E) fluctuated with SOI, this spike‐like southward movement of the salinity front was probably affected by reduction of low‐salinity water during El Niño in the north‐western Pacific Ocean. However, ENSO only induced such large southward movements of the salinity front when the time lag between the low precipitation and low SOI was short (within four months). This salinity front is quite important for long‐distance migrating fish such as the Japanese eel because the eels spawn just south of the salinity front in the NEC. This behaviour suggests that the movement of the salinity front associated with ENSO may control the success of larval transport from the spawning ground in the NEC to the nursery ground in East Asia. In fact, catch of the Japanese eel larvae in Japan was well correlated with fluctuation of SOI and the location of the salinity front, and lower catch occurred during El Niño. The salinity front has moved from 13°N to 17°N during the past three decades. Considering that conditions of larval transport are worse north of 15°N, we suggest that decadal‐scale linear decrease of glass eel catch during the past three decades also can be explained by the displacement of the salinity front.     

Changes in serum vitellogenin levels and immunohistochemical localization of vitellogenin in hepatic cells during ovarian development in the Japanese eel

ABSTRACT: The present study was designed to quantify changes in serum vitellogenin (VTG) levels and to localize VTG in hepatic cells immunohistochemically during artificial maturation of the Japanese eel Anguilla japonica . Hormone treatment resulted in a gradual rise in serum VTG levels, in which the highest values were attained at the migratory nucleus stage. Hepatocytes responded to hormone treatment by increased immunoreactivity; both staining area and intensity were enhanced with ongoing gonadal development. Moreover, cytoplasmic vacuoles increased in number during artificial maturation, occupying large areas inside hepatocytes. The unusual appearance of hepatocytes may reflect the effect of artificial maturation.     

Liver of the fish Gymnotus inaequilabiatus and nematode larvae infection: Histochemical features and expression of proliferative cell nuclear antigen

Histopathological lesions due to third‐larval stage of nematode Brevimulticaecum sp. within the liver of a subpopulation of 31 Gymnotus inaequilabiatus from the Pantanal Region (Brazil) were studied with histochemical and immunohistochemical methods. In 93.5% of fish, livers harboured nematode larvae and the intensity of infection ranged from 8 to 293. In livers with highest number of larvae, the hepatic tissue was occupied primarily by the nematodes. Each larva was encircled by focal inflammatory granulomatous reaction. Within the thickness of the granuloma, three concentric layers were recognized: an inner layer of densely packed epithelioid cells, a middle layer of mast cells (MCs) entrapped in a thin fibroblast‐connective mesh and an outer layer of fibrous connective tissue with fibroblasts. Epithelioid cells and fibroblasts within the thickness of the granuloma wall were positive for proliferative cell nuclear antigen (PCNA). Moreover, several hepatocytes in infected liver were immunoreactive to PCNA. Occurrence of rodlet cells and MCs in parenchyma, in close proximity to the encysted nematode larvae and near the blood vessel of infected liver, was observed. Macrophage aggregates (MAs) were numerous within the granulomas and scattered in parenchyma of the infected liver. High quantity of haemosiderin was encountered in MAs and hepatocytes of infected liver.     

溶解氧对团头鲂耐低氧新品系F5代的鳃组织形态及各组织酶活性的影响

为研究不同浓度溶解氧(DO)对团头鲂(Megalobrama amblycephala)耐低氧新品系F5代鳃组织形态以及各组织酶活性的影响,本研究将团头鲂在低氧[DO为(1.7±0.2)mg/L]和高氧[DO为(19.3±0.5)mg/L]条件下分别处理0、4和7d,恢复常氧[DO为(7.8±0.3)mg/L]7d后,...     

European eel distribution and body condition in a river floodplain: effect of longitudinal and lateral connectivity

Abstract – We studied eel population characteristics (size classes, densities and body condition) in the lower Loire River floodplain (France) to evaluate the effects of longitudinal and lateral gradients. A total of 36 sites were electrofished in June 2005. The sites were grouped first into three river segments according to the distance inland corresponding to a longitudinal gradient and secondly into three lateral connectivity categories. Results indicate that small eels, especially those ≤300 mm, were very abundant downstream but density rapidly decreased upstream. In addition, eels ≤150 mm tended to be most abundant in connected waterbodies. Conversely, the distribution of larger eels was quite uniform across both dimensions. Eel condition decreased upstream. In parallel, in downstream river segment, eels had a lower condition in disconnected waterbodies than in connected ones. Eel specialists still have only a longitudinal perception of eel habitat. Our results suggest that lateral gradients should be also taken into consideration.     

The influence of phytoestrogens and oestradiol-17β on growth and sex determination in the European eel (Anguilla anguilla)

Under aquaculture conditions, European eels (Anguilla anguilla) produce a high percentage of males (80–95%) that normally stop growing at 100–200 g. Females continue to grow to 500–750 g and obtain higher market value. Therefore, increasing the percentage of females in a population would be beneficial to the culture of eels. The present study was carried out in order to examine the effect of oestradiol and phytooestrogens on sex differentiation and growth rate of eels. Juvenile European eels with undifferentiated gonads were fed pellets containing oestradiol‐17β (E2) or phytooestrogens for 100–150 days. Feeding E2 resulted in 50–61% increase in body weight compared with the control. Oestradiol‐17β and phytooestrogens both elevated significantly the percentage of females in the population. Feeding E2 at 20 mg kg^?1 feed resulted in 70% females, while lower concentration of E2 (2 mg kg^?1) resulted in only 30% after 100 days (Experiment 1). The same dose given for 150 days (Experiment 2) resulted in 88% females, indicating that both, the concentration and duration of E2 treatments had a significant effect on sex differentiation. Fish fed genistein at 2 mg kg^?1 for 100 days, resulted in 55% of females, but at a higher dose of 20 mg kg^?1 there were only 15% females. These results demonstrate that phytooestrogens can be used as alternatives to gonadal steroids for sex manipulation in eels, but the optimal concentrations and duration are still to be determined.     

The influence of diel vertical migration on zooplankton transport and recruitment in an upwelling region: estimates from a coupled behavioral-physical model

Diel vertical migration (DVM) is a common zooplankton behavior in which organisms reside in surface or near‐surface waters at night and at deeper depths during the day. In many upwelling regions, DVM reduces the transport of organisms away from the region. It is unclear, however, what role DVM plays in recruitment (the arrival of larvae or juveniles to locations where they will become reproducing adults) to upwelling regions. In this study, we estimate the influence of DVM on zooplankton transport, the level of recruitment of locally produced propagules (self‐recruitment), and sources of recruits in the upwelling region near Monterey Bay, California, by simulating the trajectories of fixed‐depth and vertically migrating organisms with a drifter‐tracking algorithm driven by climatological velocity fields from a three‐dimensional hydrodynamic model. Our simulations suggest that DVM into subsurface poleward and onshore currents during the day does not fully compensate for equatorward and offshore transport in the surface Ekman layer at night and does not retain zooplankton in the Monterey Bay region. Our simulations also suggest that DVM decreases the ability of zooplankton to return to the region after being transported away and shifts source regions for recruits closer to the bay. While DVM does not appear to substantially increase the potential for self‐recruitment to the region, this study indicates that other mechanisms, such as transport during non‐upwelling periods, continuous transport below the surface, increases in mean transport depth over time, or seasonal changes in hydrography, may still enable relatively high levels of self‐recruitment to this highly advective region.     

不同蛋白水平饲料对斑节对虾家系生长及生长相关基因mRNA表达量的影响

研究了选育出的生长速度不同的9个斑节对虾(Penaeus monodon)家系在蛋白含量为34%、38%和42%饲料的饲喂下,其生长、生长激素基因(GH)、组织蛋白酶-L(Cathepsin-L)、胰蛋白酶(Trypsin)等基因的表达量及生长和基因表达量之间的相互关系。结果表明:1)斑节对虾的生长、3种基因的表达量与饲料中蛋白质含量均呈显著的正相关关系(P0.05);2)随着饲料蛋白质含量的增加,不同斑节对虾家系之间的生长差异和3种基因表达量的差异减小,相同斑节对虾家系在不同蛋白含量饲料的饲喂下,其特定生长率和基因表达量均具有显著性的差异(P0.05);3)选育的9个家系斑节对虾的生长在家系和蛋白质含量这两个因素间没有显著性的交互作用(P0.05),但是肝胰腺和肠道中3种基因的表达量在家系和蛋白质含量这两个因素间具有极显著的交互作用(P0.01),其中,饲料蛋白质含量因素对GH和Trypsin两个基因表达量的贡献率是家系因素的1.31~3.66倍,而饲料蛋白质含量因素对Cathepsin-L基因表达量的贡献率是家系因素的0.06~0.54倍。     

Dietary lipid levels affect lipoprotein clearance,fatty acid transport,lipogenesis and lipolysis at the transcriptional level in muscle and adipose tissue of large yellow croaker (Larimichthys crocea)

To investigate the effect of dietary lipid levels on lipid metabolism in muscle and adipose tissue of large yellow croaker (Larimichthys crocea), fish (average body weight 150.0 g) were fed to one of three diets with a low (6%), moderate (12%, the control diet) or high (18%) crude lipid level for 10 weeks. The expression of genes related to lipoprotein clearance (lpl, ldlr, lrp1 and srbi), fatty acid transport (cd36, fatp1, fabp3 and fabp11), lipogenesis (fas and dgat2) and lipolysis (atgl, cpt1 and aco) in muscle and adipose tissue was analysed. In muscle of fish fed the low‐lipid diet, the expression of lpl, lrp1, fas, dgat2 and atgl was significantly downregulated, whereas fabp3 and fabp11 were significantly upregulated compared with those fed the control diet. In muscle of fish fed the high‐lipid diet, the expression of lpl, ldlr, lrp1, cd36, fabp3, fas and dgat2 was significantly decreased, while fabp11 expression was significantly increased compared with those fed the control diet. Compared with the control group, only a significant downregulation of fabp3 expression in adipose tissue was observed in the low‐lipid group. In the high‐lipid group compared with the control group, the expression of ldlr, fabp11, dgat2, atgl and aco in adipose tissue was significantly upregulated, whereas cpt1 expression in adipose tissue was significantly downregulated. These results indicated that the expression of the selected key genes related to lipid metabolism was tissue‐specific regulation by dietary lipid levels in large yellow croaker.     

运输时间和MS-222浓度对翘嘴鲌皮质醇、乳酸及氧气袋内水质的影响

釆用MS-222浓度为0(C0组)、5 mg/L(C1组)、10 mg/L(C2组)、15 mg/L(C3组)、20 mg/L(C4组),运输时间为0、2、4、6、8、10 h,运输密度为33 g/L,采用5×6双因素实验,氧气袋运输翘嘴鲌(Culter alburnus)稚鱼,研究了运输时间和MS-222浓度对翘嘴鲌全鱼皮质醇、乳酸含量及氧气袋内水质的影响。结果显示:翘嘴鲌全鱼皮质醇、乳酸水平随运输时间呈现先升高后降低的趋势,随麻醉剂浓度的增加而显著降低;C1-C4组的DO、p H随运输时间呈现先降低后升高的趋势,C0组则随运输时间的增加显著降低,各组的DO和p H随MS-222浓度的增加而显著增加;C1-C4组的氨氮(TAN)和游离CO_2浓度随运输时间呈现先升高后降低的趋势,而C0组则随运输时间的增加显著升高,各组的TAN和游离CO_2浓度随MS-222浓度的增加而显著降低。时间和浓度的交互作用都显著。运输时间T6 h时,MS-222刺激翘嘴鲌,不利于运输,运输时间T6 h时,MS-222能显著降低其应激水平及代谢强度,其中C4组皮质醇、乳酸含量最低,C3、C4组水质条件最好,其适宜的麻醉剂浓度为15~20 mg/L。     

Effects of dietary levels of 20:5n-3 and 22:6n-3 on tissue lipid composition in juvenile Atlantic salmon, Salmo salar, with emphasis on brain and eye

Four dietary groups of juvenile Atlantic salmon, Salmo salar L., each with three replicates, were fed diets with increasing levels of docosahexaenoic acid (22:6n-3; DHA) and eicosapentaenoic acid (20:5n-3; EPA). Fatty acid composition of brain and eye was determined at the start and approximately every 3 weeks during the experimental period, and fatty acid composition of liver and fillet was determined in fish from the final sampling. Lipid class composition of brain and eye, and fatty acid composition of these lipid classes was determined at the end of the experiment. There was no effect of increasing dietary DHA content on fatty acid composition, lipid class composition or DHA levels in the lipid classes in the juvenile Atlantic salmon brain. The increasing dietary EPA content, however, was reflected in both the total fatty acid composition and in the EPA content in neutral lipids, phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI). A minor effect of the increasing dietary DHA content was found in the lipid composition of the juvenile salmon eye. Both EPA and 18:2n-6 levels in eye, however, clearly reflected the increasing and decreasing, respectively, dietary levels of these two fatty acids. The dietary EPA levels also affected the EPA levels in neutral lipids, PC, PE, PI and PS (phosphatidylserine) in the juvenile salmon eye. The results demonstrate that these dietary levels of DHA had no effect on brain lipid composition and only a minor effect on eye lipid composition. Furthermore, the dietary EPA levels significantly affected the lipid composition of both brain and eye. The fillet fatty acid composition reflected the dietary fatty acid composition, except for the DHA/EPA ratio, which was reversed in fillet compared with that in the diets. The liver fatty acid composition was also affected by the increasing dietary EPA and DHA levels.     

The effects of a sudden salinity change on cortisol, glucose, lactate, and osmolality levels in grouper Epinephelus malabaricus

Grouper Epinephelus malabaricus (weighing 46.37?±?5.10?g) previously maintained in 24‰ seawater were transferred to 14, 19, 24 (control), 29, and 34‰ seawater. Serum cortisol, glucose, lactate, and osmolality levels were measured at 7 time points during 240?min. Serum cortisol and glucose levels of fish transferred to 29 and 34‰ seawater significantly increased to the highest after 10 and 20?min, respectively. No significant differences in serum cortisol and glucose levels were observed for the fish after 30?min among all treatments. Serum lactate level of fish transferred to 14, 19, 29, and 34‰ seawater was significantly lower than that of the control fish after 10-30?min. However, no significant differences in serum lactate were observed 60?min among five treatments. The serum osmolality of the fish following 240-min transfer increased directly with salinity, whereas the osmoregulatory capacity value (medium osmolality-plasma osmolality) of the fish following 240-min transfer was inversely related to salinity. It is concluded that grouper showed strong osmoregulation in 14-34‰ seawater. Serum cortisol and glucose levels of fish transferred to 29 and 34‰ seawater increased rapidly in 10-30?min, indicating an early stress response.     

中华鳖NR5a2基因的克隆与表达分析

为了解NR5a2在中华鳖(Pelodiscus sinensis)性腺分化过程中的作用,通过RACE技术克隆获得中华鳖NR5a2 cDNA。结果显示:该基因序列全长为2 674 bp,开放阅读框为1 608 bp,编码535个氨基酸,5′非编码区长度为40 bp,3′非编码区长度为1 026 bp。生物信息学分析表明该基因编码的蛋白质分子质量为60.5 ku,等电点为8.57,具有保守的ZnF-C4结构域和HOLI结构域。氨基酸多重序列比对分析表明中华鳖NR5a2基因与西部锦龟(Chrysemys picta bellii)的相似性最高,其次是原鸡(Gallus gallus)。系统进化分析表明中华鳖NR5a2与西部锦龟的亲缘关系最近。实时荧光定量PCR结果表明NR5a2在中华鳖不同组织中均有表达,且卵巢组织中的表达量显著高于精巢组织。31.5℃孵化条件下,不同发育时期的中华鳖胚胎组织中发现NR5a2基因在17期时表达量达到最高,随后显著下降。结果表明NR5a2可能参与中华鳖的性别分化。     

鳗鱼及其制品中喹诺酮类药物残留的微生物快速检测方法研究

介绍了一种采用E.coliATCC8739为检测菌,检测鳗鱼中喹诺酮类药物残留的微生物抑制试验方法,该方法快速、简便,适用于大批量的样品检测。该方法的检测低限为环丙沙星20μg/kg;恩诺沙星25μg/kg;其它喹诺酮类药物50～250μg/kg,6～8h即出结果。鳗鱼及其制品的恩诺沙星的回收率为75.89%～83.12%。