投喂蚕豆对草鱼免疫器官超微结构和免疫基因表达的影响

为研究草鱼摄食蚕豆后免疫功能变化及转投30 d人工饲料后免疫增强效果,在草鱼摄食蚕豆30、60、90、120及150 d(转投饲料30 d)时,检测肠道、头肾、肝脏等器官超微结构及C-type lysozyme、IFN-I、TNF-α等免疫基因表达变化。结果表明:1)饲喂单一饲料蚕豆的草鱼,其肠道、头肾、肝脏等3个重要免疫器官受到严重损伤,转投饲料30 d后其病变并未显著好转;2)免疫基因C-type lysozyme、IFN-I、TNF-α、IgM和MHC-I的表达在30～120 d均被抑制,转投饲料30 d后,头肾中C-type lysozyme、IL-1β、IFN-I、IgM、MHC-I以及肝脏中IL-1β、TNF-α、IFN-I、IgM、MHC-I的表达量有所增加。综上,草鱼摄食蚕豆后其免疫功能受到显著抑制,而转投30 d饲料对脆肉鲩的免疫机能具有一定的恢复作用。     

Reproductive ability of second generation ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids and characteristics of their offspring

The purpose of this study was to investigate the reproductive ability of second generation (F₂) koi (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids. Only diploid F₂ males and females were fertile and used in crosses. A significant increase was recorded in male fertility in F₂ versus F₁. In contrast with an earlier study in which only one fertile F₁ male was found, about 20% of F₂ males produced sperm. The observed reproductive ability of F₂ hybrids was similar to that demonstrated by the only fertile F₁ male and F₁ females. F₂ males produced diploid spermatozoa and generated triploids when crossed with koi females. All triploid fish in these progenies were males indicating that F₂ males had a sex chromosome constitution of XY. F₂ females produced diploid eggs and generated mostly triploids when crossed with koi males. In progenies obtained by crosses of F₂ males with F₁ and F₂ females, most of the surviving juveniles (63%–100%) were diploid; a minority of juveniles were aneuploid (ploidy ranged from 2.1n to 3.6n). Diploid fish in these progenies were presumably the result of spontaneous androgenesis and gynogenesis, by the same mechanisms observed earlier in progenies obtained by crossing the F₁ male with F₁ females.     

Interactive effects of coffee bean supplementation and waterborne zinc toxicity on growth performance,biochemical variables,antioxidant activity and zinc bioaccumulation in whole body of common carp,Cyprinus carpio L.

Roasted coffee powder (RCP; Coffea arabica) is usually used as a beverage for human but there are few attempts to use it as a natural feed supplement in fish diets. In this study, common carp, Cyprinus carpio L., (11.8 ± 0.09 g) were reared in zinc (Zn)‐containing water at concentrations of 0.0 or 5.0 mg/L and cosupplemented with 0.0 or 1.0 g RCP/kg diet for 6 weeks to investigate effects of RCP supplementation, Zn exposure and their interaction on fish performance, biochemical variables, antioxidant activity and Zn bioaccumulation in whole fish body. Fish growth and feed intake were significantly affected by RCP supplementation, Zn toxicity and their interaction. However, fish fed a RCP‐supplemented diet did not exhibit better performance than those fed the RCP‐free diet and both diets produced higher fish performance than the Zn‐toxicated fish. It is noticed that RCP supplementation to Zn‐toxicated fish enhanced their growth, and feed utilization as compared to Zn‐toxicated fish alone. Fish fed control and RCP‐enriched diets showed no significant differences in biochemical variables, which were significantly altered due to waterborne Zn toxicity. Moreover, Zn reduced significantly; meanwhile, RCP supplementation increased significantly superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities. Notably, Zn exposure could reduce fish growth and antioxidant activity and increase Zn deposition in whole fish body. And RCP intake could enhance the antioxidant activity exerting a protective effect against Zn toxicity, thereby reducing Zn bioaccumulation in whole fish body.     

Dietary available phosphorus requirement for juvenile gibel carp (Carassius auratus gibelio var. CASIII)

A 57‐day growth experiment was conducted with juvenile gibel carp (13.48 ± 0.10 g) in a flow‐through system to study the effect of dietary phosphorus on growth performance, body composition, nutrition utilization, phosphorus loading and enzymes activities. Seven semipurifed diets were formulated to contain 0.07 (the basal), 2.27, 5.32, 8.10, 12.06, 15.24 and 19.48 g available phosphorus/kg diet. The results showed that specific growth rate, body length and feed efficiency significantly increased in the fish fed diets containing 0.07 to 15.24 g available P/kg diet (p < .05). Ash and P content increased in fish fed diets containing 0.07–12.06 P g/kg (p < .05) and then levelled off, while moisture, crude protein and lipid had no significant difference (p > .05). The protein retention efficiency increased in the fish fed with diets 0.07–5.32 g/kg P (p < .05) and then reached a plateau. The P content in faeces was higher in fish fed diets containing 15.24 and 19.48 g available P/kg. Total P concentration in tank water increased in fish fed 0.07–12.06 g available P per kg diet (p < .05). The plasma P was higher in the fish fed with 15.24 g available P/kg diet (p < .05), triglycerides was lower in the fish fed diet containing 15.24 and 19.48 g available P/kg (p < .05), no significant differences were observed in plasma Ca, plasma glucose and calcitonin (p > .05). Based on SGR, whole body P content and FE, dietary available P requirement for juvenile gibel carp were 13.37, 13.97 g/kg and 15.06 respectively.     

The possibility of application of agglomerate elastomers (EPP) as media for biological bed in aquaculture

The aim of the study was to determine the possibility of experimental media (agglomerate elastomers EPP) application as biological media bed, which serves the purpose of water purification in recirculating aquaculture systems (RAS). RAS enables mass‐production of fish in small volume of water in a limited area. This involves the possibility of multiple usage of water during culture. However, for that purpose of maintaining proper physico‐chemical parameters, water purification from products of metabolism, especially toxic nitrogen compounds, is required. One of the simplest and most effective ways to achieve it is combining application of two types of water filtration: mechanical and biological. It is needed to study new media for biological bed with proper filling is able to purify water from toxic nitrogen compounds.     

Non‐specific immune responses and intestinal immunity of common carp (Cyprinus carpio) fed Jujube (Ziziphus jujube) fruit extract

This study investigated the effects of Jujube (Ziziphus jujube) fruit extract (JFE) as an excellent source of nutrients and phytochemicals on the innate immune responses and expression of genes involved in intestinal immunity of common carp (Cyprinus carpio) fingerlings. Three hundred healthy carps (10.78 ± 0.05 g) fingerlings were randomly allocated to 12 fibre glass tanks (300L; 15 fish per tank) and fed for 8 weeks with experimental diets which contained different levels of JFE (0%, 0.25%, 0.5%, or 1%). Thereafter, serum non‐specific immune parameters [total immunoglobulin (Ig) as well as alternative complement (ACH50) and lysozyme activity (SL)] and expression of genes involved in intestinal immunity (lyz, tnf‐alpha, il1b, il8, il10, and tgfb) were measured. The results revealed a significant elevation of Ig level in the JFE‐fed fish (p < 0.05) over the control fish. However, there was no significant difference among treated groups in case of serum total Ig. The highest levels of SL and ACH50 were observed in fish fed 1% of JFE. Nonetheless, no significant difference was noticed regarding SL and ACH50 of control and other supplemented groups (0.25% and 0.5% JFE). Also, expression of lyz, tnf‐alpha, and il1b genes increased significantly (p < 0.05) in JFE‐fed fish, in a dose‐dependent manner. On the other hand, significant (p < 0.05) down‐regulation of il110 and tgfb were noticed in treated groups. The present findings suggested that extraction of Jujube, Z. jujube fruit, possesses beneficial effects on immune responses of common carp.     

Determination of Lipid Peroxidation and Fatty Acid Compositions in the Caviars of Rainbow Trout and Carp

In this study, roes obtained from carp (Cyprinus carpio carpio) that were caught in Keban Dam Lake (Turkey) and cultured rainbow trout (Oncorhynchus mykiss) were processed as caviar, and the products were stored at + 4 C°. Lipid peroxidation and the compositions of fatty acids of these products were assessed during production and storage period. The result of fatty analyses showed that palmitic acid (16:0), stearic acid (18:0), palmitoleic acid (16:1), oleic acid (18:1), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), which are all essential fatty acids, were determined in the caviars of the roes obtained from rainbow trout and carp. According to results of chemical analyses, there were no significant differences in thiobarbituric acid reactive substances (TBARs) between rainbow trout and carp caviars (p > 0.05), but malondialdehyde (MDA) levels were found to be statistically significant (p < 0.01). It can be concluded that caviars obtained from both rainbow trout and carp roes are rich in fatty acids and take a part in a balanced and healthy diet.     

四氯乙烯和镉联合胁迫对草鱼SOD和POD活性影响

为了考察超氧化物歧化酶(SOD)和过氧化物酶(POD)是否可以作为四氯乙烯(PCE)和镉(Cd2+)联合污染生物指标,采用静水生物测试法,研究了PCE和Cd2+联合胁迫对草鱼(Ctenopharyngodon idellus)肝胰脏、肾脏和鳃组织SOD以及肝胰脏、肾脏POD的影响。结果表明:(1)PCE和Cd2+复合污染胁迫时,草鱼肝胰脏SOD随着时间变化呈现"降低-升高-降低"的变化趋势,且最高浓度组始终低于对照;(2)肾脏SOD在所有暴露浓度组随着时间变化呈现"降低-升高-降低"的变化趋势,除最低浓度组在120 h显著高于对照外,其他均显著低于对照;(3)鳃组织SOD也呈现"降低-升高-降低"的变化趋势,但始终低于对照;(4)高浓度组(1/4TU和1/2TU)草鱼肝胰脏和肾脏POD活性随着暴露时间变化呈"降低-升高-降低"的变化趋势,但肝胰脏和肾脏的POD活性始终显著低于对照(P0.05);(5)草鱼鳃组织SOD活性明显低于肝胰脏和肾脏。PCE和Cd2+分别破坏了酶的蛋白结构和诱导了鱼体细胞膜的脂质过氧化,二者联合污染比单一污染加剧了草鱼的毒性效应。     

雌核发育草鱼群体及两个普通草鱼群体的微卫星遗传分析

以草鱼(Ctenopharyngodon idellus)养殖群体(YZ)为母本,以紫外线灭活的鲤鱼精子激活草鱼卵子,冷休克抑制第二极体排出的方法诱导获得异精雌核发育草鱼群体(CH)。利用12对微卫星引物对YZ群体、YS (扬州广陵长江系家鱼原种场引进草鱼群体)群体、CH群体进行PCR扩增并分析,共检测出194个等位基因,其中75.8个有效等位基因。YZ群体、YS群体、CH群体的平均等位基因数依次为13.0、12.6、4.7;平均有效等位基因数依次是7.7、6.6、2.3;平均期望杂合度依次为0.87、0.82、0.56;平均多态信息含量依次为0.84、0.79、0.49。从每个个体在微卫星位点的纯合率看,YZ群体中个体的纯合度在0.00～0.33, YS群体r中个体的纯合度在0.00～0.42, CH群体中个体的纯合度在0.42～0.92。这表明与YZ群体和YS群体相比,CH群体的遗传多样性显著下降,并且在每个位点的纯合率CH群体均高于普通草鱼群体,表明人工诱导减数雌核发育可加速草鱼大多数基因位点的纯合,是快速建立高纯品系的有效手段。同时,本研究筛选并利用微卫星位点组合建立了雌核发育草鱼子代不同家系及其母本亲缘关系的简易、高效鉴别技术,旨在为雌核发育草鱼标记辅助育种打下基础。     

Establishment of a new cell line susceptible to Cyprinid herpesvirus 3 (CyHV‐3) and possible latency of CyHV‐3 by temperature shift in the cells

A new cell line named CCF‐K104 predominantly consisting of fibroblastic cells showed optimal growth at temperatures from 25 °C to 30 °C. Serial morphological changes in the cells induced by Cyprinid herpesvirus 3 (CyHV‐3) included cytoplasmic vacuolar formation, cell rounding and detachment. Mature virions were purified from CyHV‐3‐infected CCF‐K104 cells by sucrose gradient ultracentrifugation and had a typical herpesvirus structure on electron microscopy. Infectious CyHV‐3 was produced stably in CCF‐K104 cells over 30 viral passages. Our findings showed that CCF‐K104 is a useful cell line for isolation and productive replication of CyHV‐3. A temperature shift from 25 °C to 15 °C or 35 °C did not allow serial morphological changes as observed at 25 °C for 14 days. Under the same conditions, real‐time PCR showed that CyHV‐3 was present with low viral DNA loads, suggesting that CyHV‐3 may establish latent infection in CCF‐K104 cells. Amplification of the left and right terminal repeat sequences of the CyHV‐3 genome arranged in a head‐to‐tail manner was detected by nested PCR following an upshift in temperature from 25 °C to 35 °C. The PCR results suggested that the circular genome may represent a latent form of CyHV‐3.