Generation and characterization of aptamers against grass carp reovirus infection for the development of rapid detection assay

Grass carp reovirus (GCRV) causes devastating viral haemorrhagic disease in farmed grass carp (Ctenopharyngon idellus). As novel molecular probes, aptamers have been widely applied in rapid diagnosis and efficient therapies against virus or diseases. In this study, three single‐stranded DNA (ssDNA) aptamers were selected against GCRV‐infected CIK cells via SELEX (systematic evolution of ligands by exponential enrichment technology). Secondary structures predicted by MFOLD indicated that aptamers formed stem‐loop structures, and GVI‐11 had the lowest ΔG value of ?30.84 KJ/mol. Three aptamers could specifically recognize GCRV‐infected CIK cells, with calculated dissociation constants (Kd) of 220.86, 176.63 and 278.66 nM for aptamers GVI‐1, GVI‐7 and GVI‐11, respectively, which indicated that they could serve as specific delivery system for antiviral therapies. The targets of aptamers GVI‐1, GVI‐7 and GVI‐11 on the surface of GCRV‐infected cells could be membrane proteins, which were trypsin‐sensitive. Furthermore, FAM‐labelled aptamer GVI‐7 could be applied to detect GCRV infection in vivo. It is the first time to generate and characterize aptamers against GCRV‐infected cells. These aptamers have great potentials in development of rapid diagnosis technology and antiviral agents against GCRV infection in aquaculture.     

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.     

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.     

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.     

草鱼幼鱼肝胰脏抗氧化系统对MC-LR胁迫的响应

为了研究微囊藻毒素(MC-LR)对草鱼(Ctenopharygodon idella)幼鱼肝胰脏抗氧化防御系统的影响,本研究通过腹腔注射的方法(剂量为25、100μg MC-LR·kg-1,分别称为低剂量和高剂量),对草鱼幼鱼进行染毒,于胁迫24、48 h和72 h后分离其肝胰脏,随后采用分光光度法检测了草鱼幼鱼肝胰脏中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性;同时采用荧光定量PCR方法分析了SOD、CAT、谷胱甘肽过氧化物酶(GPx)和谷胱甘肽还原酶(GR)基因相对表达量的变化。结果显示:高剂量MC-LR诱导24 h和48 h后,草鱼幼鱼肝胰脏SOD活性显著增加(P0.05),而SOD在低剂量组中活性没有显著变化(P0.05);低剂量MC-LR对CAT活性没有显著影响(P0.05),而高剂量MC-LR诱导24 h后可使草鱼幼鱼肝胰脏CAT活性显著增加(P0.05),随后其活性又下降,但差异不显著(P0.05)。在MC-LR胁迫过程中,SOD、CAT、GPx基因表达在两个剂量组中均显著低于对照组(P0.05),而GR基因在低剂量MC-LR胁迫24 h后相对表达量显著上调(P0.05),尽管在72 h相对表达量上调,但差异不显著(P0.05),而高剂量组中GR基因在胁迫24 h和48 h后,其表达量被抑制,但不存在显著差异(P0.05)。进行抗氧化酶活性和基因表达相关性分析发现,SOD和CAT酶活性与SOD和CAT基因表达不相关。上述研究表明MC-LR对草鱼幼鱼肝胰脏抗氧化系统产生了明显的胁迫效应,但MC-LR对机体抗氧化酶活性与基因编码调控之间的相互作用机制还有待更深入的研究。     

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.     

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.     

Koi herpesvirus and carp edema virus threaten common carp aquaculture in Croatia

Common carp (Cyprinus carpio) is a very important fish species for warm-water aquaculture in Croatia. All Croatian carp farms are subjected to a surveillance programme for the presence of koi herpesvirus (KHV), causing a deadly disease called koi herpesvirus disease (KHVD). However, there is no surveillance for other viral pathogens of importance like carp edema virus (CEV), a causative agent of koi sleepy disease (KSD). During regular testing within the KHVD surveillance programme, we tested samples for CEV simultaneously. The screening indicated possible outbreaks of KHVD and KSD. During 2016, KHVD broke out in an isolated area and soon thereafter a KHV eradication programme was successfully performed. However, during 2018 and 2019, two additional mortality events occurred in lakes in the southern part of Croatia during the spring. Samples from both events tested positive for CEV. An epidemiological investigation confirmed the introduction of infected carps from an infected farm to one of the lakes. To prevent the spreading of CEV into open waters, it is of utmost importance to introduce CEV testing before fish movement or to perform regular testing of all carp farms in the country to determine CEV prevalence for the purpose of implementation of control measures.