A comparison of artificial and natural foods and their combinations in the rearing of goldfish, Carassius auratus (L.)

Intensive grow‐out of goldfish, Carassius auratus (L.), larvae and juveniles in closed systems requires the control of environmental conditions and feeding. This study investigates the use of different types of live food and combinations of live food and dry food in a series of four rearing experiments. Juvenile goldfish can be weaned from Artemia onto live food at about 24 days after the onset of feeding without causing a reduction in growth and survival. The replacement of Artemia by Daphnia at day 10 appears feasible, as growth and survival were not significantly affected. Fish fed decapsulated Artemia cysts grew better than fish fed live Artemia. Within the first 14 days, goldfish juveniles should be fed at least 155 cysts per fish per day to achieve fast growth and to minimize size variation.     

鳜三种生长抑素cDNA全长克隆、组织表达及其在脑中定位分析

利用cDNA末端快速扩增法(RACE)克隆了鳜脑中3种生长抑素前体(preprosomatostatin,PSSⅠ、PSSⅡ和PSSⅢ)cDNA全长序列。结果显示,鳜PSSⅠcDNA全长647 bp,含开放阅读框369 bp,共编码122个氨基酸,前26个氨基酸为信号肽,C端SS-14在脊椎动物中非常保守;PSSⅡcDNA全长655 bp,开放阅读框387 bp,编码128个氨基酸,前25个氨基酸为信号肽,C端带有[Tyr7,Gly10]SS-14;PSSⅢcDNA全长823 bp,开放阅读框333 bp,共编码110个氨基酸,前21个氨基酸为信号肽,C端带有[Pro2]SS-14。利用RT-PCR检测了鳜PSS mRNA的组织表达特征,3种PSS mRNA均在脑中表达,PSSⅠmRNA在其他组织中未检测到表达,PSSⅡmRNA在食道和胃中大量表达,PSSⅢmRNA在肾和性腺中大量表达。推测3种PSS除一致参与神经调控外,PSSⅡ还可参与消化作用调控,PSSⅢ参与调控生殖作用。脑组织原位杂交分析表明,PSSⅠmRNA在下丘脑和延脑中表达,PSSⅡmRNA在下丘脑中表达,PSSⅢmRNA在下丘脑、视觉盖、延脑和脊髓中表达。3种PSS均在下丘脑中表达,表明它们可能参与腺垂体激素分泌的调节;在其他脑区存在表达差异,推测它们还可作为神经递质或调节因子,发挥不同的生物学效应。     

Digestibility in goldfish fed diets with and without chromic oxide and exposed to sublethal concentrations of cadmium

Effects of three sublethal levels of cadmium (40, 120 and 260 μg L⁻¹) on apparent total dry matter (TDM) and apparent protein (PD) digestibility in goldfish, Carassius auratus (L.), over an 8-week experimental period were investigated using diets with and without chromic oxide (0.5%) marker. The %TDM and %PD were estimated based on crude fibre as the marker for the diet without Cr₂O₃, and Cr₂O₃ and crude fibre as markers for the diet with Cr₂O₃. Generally, digestibility estimations were lower in goldfish fed the diet with the Cr₂O₃ marker. Similarly, for the diet with Cr₂O₃ marker, digestibility estimates based on crude fibre were higher than those based on Cr₂O₃. In all treatments (2 diets × control and 3 cadmium levels), the weekly estimations of %TDM and %PD did not differ significantly ( P > 0.05). The effect of cadmium on weekly %TDM and %PD was not that obvious in all treatments. However, the overall mean %TDM and %PD differed significantly ( P < 0.05) between diets and between markers, at all cadmium concentrations. The present study raises doubts regarding the validity of the use of Cr₂O₃ as a marker in digestibility studies in fish, as had been recently suggested by other authors but for different reasons.     

久效磷对金鱼精巢超显微结构和特征性酶活性的影响

用0.01、0.10、1.00 mg.L-1久效磷暴露金鱼21 d,测定了精巢乳酸脱氢酶(LDH)、酸性磷酸酶(ACP)、碱性磷酸酶(ALP)的活性,并观察了精巢显微和超微结构的变化。结果表明:久效磷暴露21 d,精巢小叶基膜断裂,Leydig氏细胞水肿,小叶间质扩大,支持细胞核膜溶解,胞质内脂滴和髓磷脂象数量增多。1.00 mg.L-1久效磷造成了生殖腺指数的显著下降,对精巢发育有一定程度的延迟作用。随着久效磷暴露浓度的升高,LDH,ACP,ALP的活性逐渐下降,0.10、1.00 mg.L-1久效磷暴露组精巢LDH,ACP的活性下降显著,而各暴露组的精巢ALP活性呈现不显著下降趋势。推测久效磷可能通过损伤精巢超显微结构和降低LDH,ACP和ALP活性,干扰精巢能量代谢,造成对雄性金鱼的生殖毒性。     

Fin erosion in a feral population of goldfish, Carassius auratus (L.), exposed to bleached kraft mill effluent

Abstract. The prevalence of acute and healed fin erosion in goldfish, Carassius auratus (L.), from the Waikato River, New Zealand, peaked in the vicinity of a pulp and paper mill which utilized a kraft pulping process in association with chlorine bleaching, and decreased down the effluent gradient. About 49% of goldfish sampled from the Kopakorahi Stream. which is part of the effluent treatment system (site 1), displayed pathological signs of fin erosion, whereas about 9% of control fish from a reference site located upstream of the official effluent discharge point were similarly affected. Fin erosion was less prevalent in male than in female goldfish, and occurred with different frequencies on different fins, the pectoral (56%) and caudal fins (33%) being most affected. Both the pectoral and caudal fins also had significantly higher proportions of healed erosion rather than acute erosion. The prevalence of acute and healed fin erosion increased significantly with increasing fish length (age). Physical deformities, predominantly in the form of eroded or otherwise malformed opercula, were found in nearly 5% of fish caught at site 1, but were not observed at the reference site. Changes in processing and bleaching technology at the pulp and paper mill in question have since been carried out, and long-term studies arc planned to monitor any subsequent improvements in fish health.     

Physiological responses of pink abalone Haliotis corrugata (Gray, 1828) exposed to different combinations of temperature and salinity

Physiological responses of pink abalone Haliotis corrugata were determined under different temperature and salinity conditions. Oxygen consumption rate was not affected by temperature and salinity. Ammonium excretion of pink abalone was inversely related to salinity. The O:N ratio indicated that abalone maintained in lower salinities had an interval of 4.9–7.7, which is indicative of a protein‐dominated metabolism, whereas the O:N in 35‰ was 28.8–35.5 for both temperatures, suggesting that carbohydrates were used as energy substrate. Haemolymph osmolality of abalone exposed to 20 and 24 °C was slightly hyperiso‐osmoconformic in salinity ranges of 20–35‰. The results of this study suggested that for optimized culture, pink abalone should be cultivated at 24 °C at a salinity of 35‰.     

缢蛏Toll样受体家族基因全基因组水平鉴定及其在弧菌刺激下的表达分析

缢蛏(Sinonovacula constricta)是我国传统的四大海水养殖贝类之一,养殖过程中易受病原菌感染而造成大量死亡和经济损失。Toll样受体(Toll-like receptor, TLR)是目前研究最多的模式识别受体,在无脊椎动物的先天性免疫中发挥着重要作用。研究缢蛏TLR分子特征及其免疫功能对预防和控制病原菌感染至关重要。本研究从缢蛏基因组序列中鉴定出42个TLR基因(ScTLR),其中,33个编码典型的TLR蛋白,其他9个为TLR样蛋白。根据蛋白结构域特点,将典型的TLR蛋白分为2大类4个亚类,一类为多半胱氨酸簇TLR (mccTLR),包括1个P型TLR和7个sPP型TLR;另一类为单半胱氨酸簇TLR (sccTLR),包括16个sP型TLR和9个Ls型TLR。与其他9种软体动物TLR基因的类型和数目比较结果显示,缢蛏基因组中未发现其他软体动物中存在的V型和Twin-TIR型TLR,起源古老的mccTLR类群在软体动物中没有大规模的扩张,而起源较晚的sccTLR类群却发生了大规模的扩张。荧光定量PCR分析显示,6种ScTLR在检测的7种组织中普遍表达,且在血细胞、鳃和肝胰腺组织中高表达。最后,副溶血弧菌(Vibrio parahemolyticus)胁迫12 h和48 h的缢蛏鳃和肝胰腺转录组数据分析显示,副溶血弧菌感染前后9个TLR基因在鳃或肝胰腺中的表达量发生显著变化,6个基因(ctg118.25、ctg118.26、ctg356.25、ctg774.6、ctg681.6和ctg1513.5)在感染12 h或48 h后的鳃组织中表达差异显著,前5个基因表达量上调,仅ctg1513.5表达量下调;3个基因(ctg467.9、ctg2496.3和ctg903.17)在感染12 h或48 h后的肝胰腺中表达差异显著,其中,ctg467.9和ctg2496.3表达量下调,ctg903.17表达量上调。综上所述,本研究结果将为深入探讨不同TLR基因在缢蛏天然免疫中发挥的作用提供研究基础。     

Isolation, sequence analysis, and characterization of androgen receptor in Southern catfish, Silurus meridionalis

Androgen receptor (AR), the mediator of androgen, plays important roles in the androgen signal pathway. In the present study, we isolated and analyzed the cDNA sequence and tissue distribution of androgen receptor in Southern catfish (scAR). The full-length cDNA of scAR contains 3,116 bp with an open reading frame (ORF) of 2,415 bp, encoding a protein of 804 amino acids (aa). Tissue distribution analysis of scAR revealed that it was expressed in all tissues examined, with no sexual dimorphism in the ovary and testis. Phylogenetic analysis and multiple amino acids sequence alignment indicated the close relationship and high similarity of scAR with ARs from cypriniform species. In addition, partial sequences of ARs from 7 other siluriform species were also isolated. Comparison of catfish ARs with those from other vertebrates revealed that an extra C-terminal tail of about 20aa exists in all the ARs from siluriform fishes investigated, but not in other ARs. The extra sequence was resulted from a 4-bp insertion before the stop codon of other vertebrate ARs, and it was identical in ARs from siluriform species of the same genus but different among ARs from species of different genera. We report here for the first time that the ARs from siluriform species are longer in C-terminal than those from other vertebrates and it might be useful in reconstruction of the phylogenetic relationship among siluriform fishes. The significance of the extra C-terminal tail for AR function remains elusive.     

Molecular cloning, characterization, and gene expression of the androgen receptor in the large yellow croaker, Larimichthys crocea

Androgens mediate a wide range of physiological responses and developmental processes in vertebrates, involving both reproductive and nonreproductive systems. The activity of androgens is mediated by the androgen receptor (AR), a member of the nuclear receptor superfamily. In this study, an AR gene was cloned from the large yellow croaker (Larimichthys crocea) for the first time. qRT-PCR revealed ubiquitous expression of AR in all adult tissues examined, with higher expression in the gonad and liver of both sexes and highest expression in the blastula stage of embryonic development. Using in situ hybridization, we detected positive signals of AR in the spermatogonium, spermatocyte, spermatid, and spermatozoon during spermatogenesis, in the cytoplasm of all oocytes during oogenesis and in the follicle cells of stage IV oocytes. Our findings support the important role that AR plays in gametogenesis, gonadal development, and the early stages of embryonic development.     

小体鲟AR基因克隆、序列分析及其组织mRNA表达

采用RT-PCR 和RACE技术, 克隆了小体鲟(Acipenser ruthenus)雄激素受体(AR)基因cDNA全长序列, 应用real-time qPCR法对雌、雄个体不同组织中AR mRNA 的表达进行检测。序列分析表明, AR基因cDNA全长为2 953 bp, 包括5′端68 bp的非编码区(untranslation region, UTR), 2 528 bp的开放阅读框ORF(open reading frame)和3′端327 bp的非编码区(不包括ploy A 的尾巴)。AR前体蛋白由843个氨基酸组成, 理论分子质量为94.24 kD。同源比较结果表明, 小体鲟AR氨基酸序列与其他鱼类的AR氨基酸序列同源性较高, 与杂交鲟(A. ruthenus × Huso huso)、西伯利亚鲟(A. baerii)、虹鳟(Oncorhynchus mykiss)和日本鳗鲡(Anguilla japonica)的雄激素受体同源性分别为99%、98%、83%和83%。real-time qPCR分析小体鲟雌雄个体不同组织中的表达结果显示, AR基因在雌雄个体中相对表达基本相同, 都是在性腺、肌肉和肾中高量表达, 而在其他组织中少量表达。AR基因在小体鲟体内广泛表达, 表明该基因可能对其生长和繁殖有重要作用。

     

野生石鲽精巢发育及注射LHRH-A对血浆T水平的影响研究

采用常规石蜡切片，组织染色法和放射免疫测定方法（RIA）对石鲽（Kareius bicoloratus）精巢的发育状况和生殖季节血浆睾酮（T）对注射外源促黄体激素释放激素类似物（LHRH-A）的反应进行研究。结果表明，石鲽精巢为小叶型，精巢发育分为6个时期。性腺成熟系数（GSI），肝重指数（HSI）和肥满度（CF）呈现显著的季节变化。注射LHRH-A后6h,T含量显著上升，之后的12h、24h逐渐下降，48h又显著回升，几乎达到注射前的8倍；对照组血浆T含量在注射后6h显著下降，12h变化不大；24h又显著回升，48h继续回升，但低于处理组。     

Changes in ovary and testis and breeding fitness of the climbing perch,Anabas testudineus (Bloch, 1792), exposed to sub‐lethal concentrations of monocrotophos

Monocrotophos (MCP) is one of the organophosphate pesticides extensively used in agriculture and animal husbandry. The present study reports the effects of sub‐lethal concentrations of MCP on gonadosomatic index (GSI), gonadal histology and breeding fitness of the fish Anabas testudineus. The fish were exposed to three sub‐lethal concentrations of monocrotophos (T1: 3.5 mg/L, T2: 5.3 mg/L and T3: 10.6 mg/L) for 45 days with 12 fish (males and females in equal numbers) in each tank, and one group was kept as control (C) without any treatment. GSI decreased in all treatment groups in comparison with control. Significant decrease in fertilization and hatching rate was observed at all concentrations in comparison with control on 15, 30 and 45 days of monocrotophos exposure. The decreased vitellogenesis, disruption of follicular wall and oocyte atresia were observed in all the treated females at 30 and 45 days of exposure. Rupture of seminiferous tubules was noticed in all the treated males irrespective of exposure periods. The results thus indicated that monocrotophos could deleteriously impact the gonadal structure and function in A. testudineus, which could seriously impact the reproductive success of the animal.     

Plasma proteins,hepatic enzymes,thyroid hormones and liver histopathology of Cyprinus carpio (Linnaeus, 1758) exposed to an oxadiazin pesticide,indoxacarb

Indoxacarb is a relatively new pesticide from oxadiazin class, which is used near carp ponds for agricultural purposes. Thus, the aim of this study was to determine indoxacarb effects on common carp (Cyprinus carpio). The fish were exposed to 0 (control), 0.75, 1.5 and 3 ppm of indoxacarb over 21 days and plasma biochemical characteristics and liver histopathology were examined. Exposure to indoxacarb induced fall in total protein after 21 days. Globulin increased after 7 days and then decreased after 14‐ and 21‐day exposure. Plasma alanine transaminase and aspartate transaminase activities increased in 1.5 and 3 ppm treatments after 7‐day exposure. Indoxacarb exposure significantly decreased plasma alkaline phosphatase after 7 days with no change at the second and third samplings. After 7 days, plasma T₃ levels had no significant change; however, it decreased after 14 days in the 1.5 and 3 ppm treatments and also reduced after 21‐day exposure to the 3 ppm treatment compared to the control group. Plasma T₄ level only decreased significantly in the 3 ppm treatment compared to the control group after 21 days. Different histopathological symptoms such as necrosis, hyperaemia, sinusoidal space extension, pyknotic nuclei, leucocyte infiltration and melanomacrophage aggregates were observed after 21‐day exposure to indoxacarb. The symptoms intensity was dependent on indoxacarb concentration. In conclusion, the present results show that indoxacarb exposure adversely affects common carp health and welfare, which consequently may induce serious problems in this species aquaculture.     

The stimulatory effect of LED light spectra on genes related to photoreceptors and skin pigmentation in goldfish (Carassius auratus)

This study aimed to assess differences in genes related to skin color of goldfish (Carassius auratus) exposed to light-emitting diodes (LEDs): red, green, and purple. We investigated differences in the expression of mammalian-like melanopsin (Opn4m), rhodopsin (RH), melanin-concentrating hormone (MCH), melanin-concentrating hormone receptor (MCH-R), and proopiomelanocortin (POMC) in goldfish exposed to different LED light spectra. Opn4m, RH, MCH, and MCH-R mRNA levels were significantly higher in the green and purple LED groups than in the white fluorescent bulb (control) and red LED groups. Furthermore, skin cells were isolated to measure the MCH-R mRNA expression levels. The results show that the mRNA expression levels were significantly higher in the green and purple LED groups than in the control and red LED groups. In addition, body weights in the green and purple LED groups were significantly higher than those in the control and red LED groups. However, POMC mRNA expression levels in the green and purple LED groups were significantly lower than those in the control and red LED groups. These results suggest that specific wavelengths regulate fish skin color through neuropeptide hormones and photoreceptors, and POMC, which is related to stress hormones and melatonin, is associated with stress levels as well as skin color.     

Histopathological changes in the liver and brain of fish exposed to endosulfan insecticide during tsetse fly control operations in Botswana

Abstract. During tsetse fly control operations, fish in the Okavango Delta region of Botswana are exposed to ultra–low–volume doses (6–12 g/ha) of endosulfan, an insecticide recognized to be highly toxic to fish. Approximately 1 % of the fish population can be killed by a spray, and the physiology and behaviour of survivors is temporarily disturbed. Simultaneously with the onset of the spraying season (July), the livers and brains of several species [including Clarias gariepinus (Burchell) and Tilapia rendalli (Boul.)] developed pathological lesions which often persisted until the season ended (October). Initially, hepatic lesions in C. gariepinus were characterized by generalized toxic necrosis, focal necrosis, and subcapsular oedema which gradually lessened in intensity and disappeared before the spraying season ended. Melano-macrophage centres and perivascular haemopoietic elements were lost from poisoned livers and were not replaced until at least 6 months later. In T. rendalli brain, pesticide–related changes included encephalitis, meningitis and oedema, with an associated inflammatory infiltrate of eosinophilic granule cells. Most brain damage disappeared during the month after the spraying season, although glial scarring was occasionally observed in fish sampled later. The possible connection of these abnormalities with observed behavioural changes is discussed.