Effects of graded levels of dietary myo‐inositol on non‐specific immune and specific immune parameters in juvenile Jian carp (Cyprinus carpio var. Jian)

The aim of this study was to evaluate the effects of myo‐inositol (MI) on non‐specific immune and specific immune defence in fish. A total of 1050 Jian carp (Cyprinus carpio var. Jian) (22.28±0.07 g) were randomly distributed into seven groups, of three replicates each, of feeding diets containing graded levels of MI (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg MI kg^?1 diet). After a 60‐day growth trial, an infectious trial was conducted by injection of Aeromonas hydrophila for 17 days. Results showed that the red blood cell (RBC) and the white blood cell count were significantly increased with increasing MI levels up to 535.8 mg kg^?1 diet (P<0.05). The spleen index showed a tendency similar to RBC, whereas the head kidney index showed the inverse pattern (P<0.05). The phagocytic activity of leucocytes, haemagglutination titre, lysozyme activity, anti‐A. hydrophila antibody titre and immunoglobulin M, after being injected with A. hydrophila, were all improved with an increase in the MI levels up to 232.7–687.3 mg kg^?1 diet respectively (P<0.05). Myo‐inositol did not influence serum acid phosphatase activity and total iron‐binding capacity (P>0.05). These results suggested that MI could enhance non‐specific immune and specific immune responses in fish.     

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.     

Effects of dietary zinc sources and levels on growth performance,tissue zinc retention and antioxidant response of juvenile common carp (Cyprinus carpio var. Jian) fed diets containing phytic acid

Plant protein‐based diets are widely used for common carp (Cyprinus carpio var. Jian). The plant source antagonist, such as phytate, negatively affects the bioavailability of trace minerals, but this aspect was not considered in previous studies on this species. A 10‐week growth trial was conducted to investigate the effects of inorganic Zinc (ZnSO₄·7H₂O, Zn‐S) and organic Zn (2‐hydroxy‐4‐(methylthio) butanoic Zinc, Zn‐M) on growth performance, Zn retention and antioxidant responses of common carp (initial weight 10.0 ± 0.03 g). Eleven semi‐purified diets containing 7 mg/kg phytic acid were prepared with 0, 5, 20, 35, 50 and 65 mg/kg Zn from Zn‐S and Zn‐M (12.8 mg/kg Zn in basal level), respectively. The results showed that either Zn‐S or Zn‐M inclusion improved growth performance, vertebral Zn deposition and antioxidant responses of fish. The Zn requirement based on WGR, vertebrae Zn and plasma SOD were 47.0, >77.8 and 48.5 mg/kg, and 63.3, 42.2 and 36.8 mg/kg, respectively, with Zn‐S and Zn‐M as Zn sources. The relative bioavailability of Zn‐M was 2.44‐ (vertebrae Zn deposition) and 1.74‐fold (plasma SOD activity) the availability of Zn‐S in the diet of common carp. The lower growth performance and poor bioavailability in vertebrae of Zn‐S group were related to the chelating of zinc with phytic acid.     

Study on the adaptability of common carp (Cyprinus carpio) to diets from the perspective of digestion and absorption

As an ecologically sustainable aquaculture mode, the rice‐fish approach has been paid more attention in recent years. In rice paddies, there are plant and animal diet items available to fish, but it is not clear how common carp adapt to different diets. In view of this, common carp (initial weight 492.6 ± 30 g, n = 270) were randomly divided into three groups and fed with earthworms (group A), earthworms + duckweed (group M) and duckweed (group P) respectively. After 8 weeks under these feeding regimes, the intestinal digestive enzymes activities were assayed, and intestinal tissue sections stained with HE and AB‐PAS were used to observe the morphology. The results showed that the activities of trypsin and lipase were highest in group A, followed by group M and group P. Group A had greater intestinal fold height, fold width and fold absorption area than the other groups (p < .05), especially in the foregut. Meanwhile, the total number of mucous cells was largest in group P, followed by the groups M and A. Totally, activities and distributions of digestive enzymes, fold height, fold width, fold absorption area and number and distribution of mucous cells of common carp were affected by the type of diet. Based on these results, it was found that common carp has strong adaptability to diets, and it was speculated that the majority of digestion and absorption of protein was concentrated in the foregut, while for starch and cellulose, it was spread out along the whole intestine.     

鲤lipea基因表达的时空特征及与脂肪沉积相关性分析

本文旨在克隆鲤(Cyprinus carpio)激素敏感性脂肪酶a (hormone-sensitive lipase a, lipea)基因,探究其时空表达特征及其与脂肪沉积的关系,以期为鲤脂肪沉积分子调控机制研究奠定理论基础。本实验采用RACE技术克隆获得鲤lipea基因,其cDNA全长为3379 bp,包括230 bp的5′非翻译区(5′-UTR)、1067 bp的3′非翻译区(3′-UTR)和编码693个氨基酸的开放阅读框。多序列比对显示,鲤lipea与斑马鱼(Danio rerio)、牙鲆(Paralichthys olivaceus)和虹鳟(Oncorhynchusmykiss)的氨基酸序列相似性分别为88.17%、64.94%和63.35%,不同物种间序列差异主要集中在C-端的多肽序列上。蛋白质结构分析显示该蛋白含有3个功能域分别具有脂肪酶活性、乙酰基水解酶活性和水解酶活性。应用实时荧光定量PCR技术分析了lipea基因在鲤各组织、不同发育时期以及脂肪含量极端差异个体的肌肉和脂肪组织中mRNA表达量的变化。结果显示, lipea在各组织中均有表达,腹腔脂肪中相对表达量最高,其次是腹部肌肉,血液中最少。随着胚胎的发育, lipea的表达水平呈现下降趋势, 8细胞期表达量最高,其次是囊胚晚期,开口期最少。lipea表达量与背部肌肉脂肪含量呈正相关,但差异不显著(P0.05),与腹腔脂肪量呈负相关,且差异极显著性(P0.01),推测lipea基因对鲤腹腔脂肪沉积可能具有抑制作用。本实验结果可为进一步解析鲤lipea基因的功能和基因调控脂肪沉积分子机制研究提供参考。     

Use of supplemental amino acids on the success of reducing dietary protein levels for Jian carp (Cyprinus carpio var. Jian)

A feeding trial was conducted to investigate the effects of essential amino acids (EAA) supplementation on the success of dietary protein reduction for Jian carp (Cyprinus carpio var. Jian). Five diets were formulated including a positive control (Diet 1, 378.4 g/kg protein), a negative control (Diet 5, 289.8 g/kg protein) and three test diets (Diet 2‐Diet 4, 340.4–299.0 g/kg protein supplemented with EAA). No significant decline of growth performance in fish fed decreased dietary protein with EAA supplementation, which were significantly higher than the negative control. The apparent digestibility coefficients (ADC) of protein and most EAAs were not significantly affected by the dietary treatments. Dietary treatments did not affect whole‐body composition, but influenced protein and EAAs retentions. Total ammonia nitrogen (TAN) excretion decreased with the decreasing dietary protein, and the highest plasma ammonia content was observed in fish fed Diet 5. Plasma glucose concentration increased with the decreasing dietary protein reduction, while other plasma parameters were not affected by dietary treatments. These results indicated that low‐protein diets (299.0 g/kg) supplemented with EAA can maintain protein, EAA digestibility and retention in Jian carp, and also has benefit in the reduction of TAN excretion.     

Effects of juniper berry oil on growth performance and blood parameters in common carp (Cyprinus carpio)

In this study the effects of commercial juniper berry oil (JBO) as a feed additive in diets for common carp (Cyprinus carpio) was investigated in terms of growth performance, feed utilization and blood parameters. The experimental diets were prepared to contain JBO at rate of 0 (JBO‐0), 5 (JBO‐5) and 10 (JBO‐10) ml/kg. Common carps (3.07 ± 0.15 g) were fed with test diets for a period of 60 days in 40‐L aquariums of triplicate groups. The feeding experiment revealed that JBO groups showed significantly better growth performance and feed utilization compared with the control group (JBO‐0) (p < 0.05). No significant effect was observed on hemoglobin amount, haematocrit ratio and consequential erythrocyte indices (p > 0.05). In the JBO groups, serum cholesterol, triglyceride, glutamic–pyruvic transaminase and alkaline phosphatase levels decreased significantly (p < 0.05). In contrast, the total protein level showed a significant increase, while glucose, albumin and lactate dehydrogenase levels did not present any significant variation (p > 0.05). Therefore, dietary addition of 5 ml/kg JBO promoted fish growth and positively influenced the blood parameters in common carps.     

Antibody response of two populations of common carp, Cyprinus carpio L., exposed to koi herpesvirus

Common carp, Cyprinus carpio L., exposed to koi herpesvirus (KHV) may become persistently infected and populations containing such virus-infected individuals may transmit the virus to other fish when co-habited. Detection of virus-infected fish in a population is thus critical to surveillance and control programmes for KHV. A study was therefore designed to detect anti-KHV serum antibodies, with an enzyme-linked immunosorbent assay, in common carp following experimental exposures to KHV under varying environmental conditions. The study determined that a proportion of fish within a population experimentally exposed to KHV (at least 10–25%) develop high antibody titres (1/1600 or greater) to the virus, and this immunological response was detectable for several months (observed at the termination of the experiments at 65, 46 and 27 weeks post-exposure). Furthermore, this response was detected in one population of fish that did not succumb to a high level of mortality when maintained at water temperatures that were non-permissive for KHV. Elevating the water temperatures to permissive conditions for KHV resulted in recurrence of disease despite the presence of anti-virus antibodies, suggesting that serum antibodies alone are not protective under the conditions of our trials.     

锦鲤腐皮病灭活疫苗制备及使用

从患有腐皮病的锦鲤(Cryprinus carpio)肝、肾、脾、血液里分离到1株优势菌,对该菌进行纯化培养并经人工感染试验,确定该菌为病原菌。经16S r RNA序列分析,鉴定该病原菌为蜡样芽孢杆菌(Bacillus cereus)。用不同浓度福尔马林和不同温度灭活蜡样芽孢杆菌,筛选其灭活效果并制得灭活疫苗。将灭活疫苗免疫健康锦鲤后测定其抗体效价变化及免疫保护率。结果表明,0.2%的福尔马林溶液在28℃下灭活24 h及4℃灭活36 h为蜡样芽胞杆菌的适宜灭活条件。首次免疫后第20天,免疫Ⅰ组、Ⅱ组、Ⅲ组、Ⅳ组血清抗蜡样芽孢杆菌的抗体效价达到高峰,分别为1∶128、1∶64、1∶64、1∶32;第30天同样剂量加强免疫后,各免疫组血清抗体效价达到更高水平,分别为1∶256、1∶256、1∶128、1∶64;免疫后第50天时攻毒,Ⅰ组、Ⅱ组、Ⅲ组、Ⅳ组免疫保护率分别为66%、66%、55%、44%。     

饲料中添加低聚木糖对鲤鱼免疫力和生长性能的影响

为探究不同质量浓度的低聚木糖调控鲤鱼免疫力及生长性能的机理,以450尾平均体质量（4．93±0．03）g的健康鲤鱼为对象,研究饲料中添加低聚木糖对鲤鱼生长性能和免疫指标的影响。以不加低聚木糖的基础饲料为对照组,试验组为分别在基础饲料中添加0．1％低聚木糖组、0．2％低聚木糖组、0．4％低聚木糖组和0．6％低聚木糖组。试验结果表明,低聚木糖可提高鲤鱼血清溶菌酶活性,但对补体旁路途径溶血活性无影响,0．6％低聚木糖组的血清溶菌酶活性显著高于对照组（P＜0．05）。对于头肾的5种免疫相关基因,低聚木糖可上调白细胞介素-1β和白细胞介素-10的基因表达,下调诱导型一氧化氮合酶和肿瘤坏死因子-α的基因表达,而对转化生长因子-β的基因表达无影响。与对照组相比,0．2％低聚木糖组、0．6％低聚木糖组的白细胞介素-1β与0．2％低聚木糖组的白细胞介素-10表达量均显著升高,所有试验组的诱导型一氧化氮合酶和肿瘤坏死因子-α的表达量均显著下降（P＜0．05）。生长性能试验结果显示,试验组鲤鱼的生长性能指标均高于对照组,其中0．6％低聚木糖组与对照组差异显著（P＜0．05）。