胰岛素受体-1在尼罗罗非鱼不同组织中的表达及其对注射葡萄糖的响应

本试验旨在研究胰岛素受体-1(IR-1)在尼罗罗非鱼不同组织中的表达及其对注射葡萄糖的响应。利用PCR扩增的方法从尼罗罗非鱼肌肉中克隆IR-1的c DNA片段,并通过半定量PCR检测,比较IR-1在肌肉、肝脏和心脏中的表达差异。选取体重约为100 g的尼罗罗非鱼160尾,随机分2组,每组4个重复,每个重复20尾。试验组腹腔注射葡萄糖(每100 g体重30 mg),对照组以相同剂量腹腔注射0.7%的无菌生理盐水。在注射前(0 h)和注射后的1、3、6和12 h分别进行采样,测定血浆葡萄糖和胰岛素含量,并通过实时荧光定量PCR检测IR-1在肌肉、心脏和肝脏中的mRNA相对表达量。结果显示:1)克隆出的IR-1 c DNA片段,其Gen Bank登陆号为JN967750,大小为1 979 bp,编码548个氨基酸。序列分析发现,尼罗罗非鱼的IR-1与其他物种比较具有很高的保守性,并具有丰富的酪氨酸激酶特征性序列。2)IR-1在尼罗罗非鱼肌肉、心脏和肝脏中均有较高的表达量,其中在肝脏和肌肉中的表达量基本一致,而在心脏中的表达量相对较低。3)试验组血浆葡萄糖含量在注射葡萄糖1 h后达到最高,并显著高于对照组(P0.05),而后开始下降,3 h后恢复到正常水平;试验组血浆胰岛素含量在葡萄糖注射3 h后达到最高,并显著高于对照组(P0.05),而后开始下降,12 h后恢复到正常水平。试验组肌肉和肝脏中IR-1 mRNA的相对表达量在注射葡萄糖后6 h时达到最高,显著高于对照组(P0.05),在12 h时恢复到正常水平;试验组心脏中IR-1 mRNA的相对表达量在注射葡萄糖后的12 h内没有发生显著变化(P0.05)。结果表明,注射葡萄糖后即刻升高了尼罗罗非鱼的血浆葡萄糖含量,相对于血浆葡萄糖含量的升高,血浆胰岛素含量的升高相对延迟,而肌肉和肝脏中IR-1 mRNA相对表达量的提高又延迟于血浆胰岛素含量的升高,从而加重了尼罗罗非鱼对葡萄糖的代谢负担。     

Salmon testes meal as a functional feed additive in fish meal and plant protein‐based diets for rainbow trout (Oncorhynchus mykiss Walbaum) and Nile tilapia (Oreochromis niloticus L.) fry

Fishery processing by‐products are a large resource from which to produce fishmeal and other products for a variety of uses. In this study, testes meal (TM) produced from pink salmon processing by‐product was evaluated as a functional ingredient in aquafeeds. Nile tilapia and rainbow trout fry were fed five isonitrogenous and isoenergetic experimental diets for 4 and 9 weeks respectively. Two diets were fishmeal‐based (FM) and three were plant protein‐based (PP). Salmon TM was added to the FM and PP diets at 7% to replace 20% of fishmeal protein (FMTM and PPTM respectively). An additional control diet was prepared in which fishmeal was added to the PP diet to supply an equivalent amount of protein as supplied by TM (PPFM). Inclusion of TM in both the FM‐ and PP‐based diets resulted in higher final body weights, although differences were only significant between rainbow trout fed FM or FMTM diets. Similar differences were calculated for other indices of fish performance, e.g. specific growth rate, feed conversion ratio, protein efficiency ratio and protein retention efficiency. Feed intake was significantly higher for fish fed FMTM compared with FM in rainbow trout. For tilapia, final weights were numerically higher, but not significantly different for fish fed diets containing TM compared with non‐TM diets (FM vs. FMTM; PP vs. PPTM). Performance of trout or tilapia fed the PPFM diet did not increase compared with the PP diet. The results indicate that TM addition to both FM and PP diets increased feed intake and also increased metabolic efficiency, demonstrating that TM can be a functional ingredient in aquafeeds.     

奥尼罗非鱼肌肉营养成分分析和营养价值评定

为了解奥尼罗非鱼肌肉营养价值,用常规方法分析奥尼罗非鱼肌肉中营养成分组成与含量。结果显示,奥尼罗非鱼肌肉中水分含量为77.90%,蛋白质为18.70%,粗脂肪为2. 65%,灰分含量为1.09%。肌肉中共检测出17种氨基酸(除色氨酸),总量为16.90%（占鲜样%）,必需氨基酸指数(EAAI)为67.68,基本符合FAO/WHO的标准。根据AAS的评分标准得出,奥尼罗非鱼的第一限制性氨基酸为苏氨酸,第二限制性氨基酸为含硫氨基酸(蛋氨酸和胱氨酸)。而CS的评分结果表明奥尼罗非鱼的第一限制性氨基酸为含硫氨基酸(蛋氨酸和胱氨酸),苏氨酸为第二限制性氨基酸。此外,鲜味氨基酸(天门冬氨酸、谷氨酸、甘氨酸和丙氨酸)含量较高占氨基酸总量的39.11%?。通过研究表明奥尼罗非鱼是一种味道鲜美、食用价值与保健作用较高的鱼类。     

Efficacy of florfenicol for control of mortality associated with Francisella noatunensis subsp. orientalis in Nile tilapia,Oreochromis niloticus (L.)

Francisella noatunensis subsp. orientalis (Fno) (syn. F. asiatica) is an emergent Gram‐negative facultative intracellular bacterium. Although it is considered one of the most pathogenic bacteria in fish, there are no commercially available treatments or vaccines. The objective of this project was to determine the most efficacious concentration of florfenicol (FFC) [10, 15 or 20 mg FFC kg^?1 body weight (bw) per days for 10 days] administered in feed to control experimentally induced infections of Fno in Nile tilapia, Oreochromis niloticus (L.), reared in a recirculating aquaculture system. The cumulative mortality of fish that received 0, 10, 15 or 20 mg FFC kg^?1 bw per day was 60, 37, 14 and 16%, respectively. Francisella noatunensis subsp. orientalis genome equivalents were detected in water from all challenged groups with slight reduction in the concentration in the florfenicol‐treated groups 4 days after treatment. The mean LOG of CFU Fno mg^?1 spleen was 3–5 and was present in all challenged groups at necropsy 11 days after treatment (21 days after challenge). Results show that florfenicol administered at doses of 15 and 20 mg FFC kg^?1 bw per days for 10 days significantly reduced mortality associated with francisellosis in Nile tilapia.     

低温对罗非鱼基因组DNA甲基化的影响

以经过连续多代抗寒选育获得的尼罗罗非鱼耐寒品系为实验材料,采用DNA甲基化敏感扩增多态性(methylation sensitive amplified polymorphism,MSAP)分析方法从全基因组水平上探讨了低温适应对罗非鱼DNA序列中CCGG位点的甲基化水平的影响。结果显示,选用的18对选择性扩增引物共检测到849个位点。耐寒品系检测到的位点数为411个,对照组为438个,其中发生甲基化的位点数分别为72和104个,总甲基化率分别为17.52%和23.74%;全甲基化位点分别为37个和65个,全甲基化率分别为9.00%和14.84%;半甲基化位点分别为35个和39个,半甲基化率分别为8.52%和8.90%。结果分析表明,尼罗罗非鱼耐寒品系的总甲基化水平、全甲基化水平和半甲基化水平较对照组均有一定程度的下降。与对照组相比,尼罗罗非鱼耐寒品系基因组DNA甲基化总体水平下降了6.22%,其中以全甲基化位点变异为主,其下降幅度比例明显,为5.84%。由此可见,经过连续多代的低温胁迫可导致尼罗罗非鱼DNA甲基化水平发生改变,发生了去甲基化反应,表现为基因组甲基化程度降低的特征,说明了DNA甲基化与罗非鱼抗寒性反应密切相关。     

Relationship Between Dietary Protein Source and Feeding Frequency During Feeding Nile Tilapia,Oreochromis niloticus (L.) Cultured in Concrete Tanks

ABSTRACT

A 24-week feeding trial was conducted in concrete tanks with Nile tilapia, Oreochromis niloticus (L.) with an average initial weight and length of 50.87 ± 6.03 g and 14.4 ± 0.45 cm, respectively, to examine the effect of two protein sources and two feeding frequencies on growth performance, production traits, and body composition. Twelve 4 m³ concrete tanks (2 × 2 × 1.25 m, long, width, and height) were each stocked with 100 fish and fed diets containing either fish meal protein or soybean meal protein at two feeding frequency of two times daily or four times daily (2 × 2 × 3 factorial experiment). The results revealed that there was no significant effect of dietary protein sources on growth rate, whereas there was a significant increase in growth rate with increasing feeding frequency (P < 0.05). The same trend was also observed for mean body weight (g), production rate (kg/m3), specific growth rate (SGR % day^?1). The best final mean body weight (g), specific growth rate (SGR % day^?1), and production rate (kg/m3) were recorded in groups of fish fed with feeding frequency four times daily. Whole fish moisture, protein, fat, and ash contents were significantly influenced (P ≤ 0.05) by protein sources and feeding frequency, except protein not influenced with feeding frequency. Energy was significantly (P < 0.05) influenced by feeding frequency, but not by dietary protein sources. Economic evaluation indicated that soybean meal (Diet B) at four times feeding daily was the most cost-effective and affordable feed strategy for farmers. We conclude that a soybean diet fed four times daily is recommended for adult Nile tilapia reared in concrete tanks.     

Natural Food Selectivity Changes with Weights of Nile Tilapia,Oreochromis niloticus (Linnaeus), Reared in Fertilized Earthen Ponds

This work was carried out to study the relationship between fish weight and natural food selectivity of Nile tilapia, Oreochromis niloticus (Linnaeus) reared in earthen ponds without supplementary diet and to guide the adaptation of feeding and fertilization strategies. Fish were stocked (1.5 fish/m²) in four fertilized earthen ponds (0.1 ha) for 6 months. Fifty fish were sampled monthly to estimate individual total weight and length and evaluate stomach contents. Fish specimens were categorized into different weight classes: <25 g, 25–50 g, 50–75 g, 75–100 g, and 100–125 g. Overall, the 50–75 g weight class had the highest degree of stomach fullness. The intestinal length-fish length ratio was similar across all fish weights, ranging from 2.9–3.4. Likewise, the composition of food items found in fish stomach was ranked as phytoplankton > detritus > zooplankton. Phytoplankton contribution to gut content increased with increasing fish weight, while detritus and zooplankton contribution decreased. Zooplankton never exceeded 1% of total stomach contents. Cyanobacteria, Chlorophyceae, Bacillariophyceae and Euglenophyceae mostly represented the phytoplankton. Bacillariophyceae dominated the phytoplanktonic portion of stomach contents at small fish size (<75 g/fish), while Chlorophyceae is the dominant group at large fish size (> 75 g/fish). Fish could select Cyanobacteria and Euglenophyceae at all fish weights, meanwhile Chlorophyceae and Bacillariophyceae were eaten with slight selectivity at larger weights..     

Effects of Yeast Oligosaccharide Diet Supplements on Growth and Disease Resistance in Juvenile Nile Tilapia,Oreochromis niloticus

Commercially available yeast and yeast subcomponents consisting mainly of β-glucan or oligosaccharide feed additives were added to diets of juvenile (12–18g) Nile tilapia (Oreochromis niloticus) at rates recommended by suppliers. Three experiments were conducted following a basic protocol with varied rates of supplementation, duration of feeding, and stocking densities. Experimental diets were fed twice daily to apparent satiation for a period of two or four weeks, at the end of which feed consumption and weight gain were measured. Following the experimental feeding period, serum components, including protein and immunoglobulin concentrations, as well as lysozyme and complement activities, were measured. A disease challenge was conducted with pathogenic isolates of Streptococcus iniae or Edwardsiella tarda. Weight gains were not significantly different in fish fed the supplemented diets when compared to the control diet. There were significant differences in fed intake within individual experiments; however, this effect was not consistent in all three experiments. Overall feed efficiency was not significantly affected by diet. There were no differences in serum components of fish sampled at two or four weeks. Fish fed the experimental diets did not have lower mortality or morbidity after disease challenge compared to fish fed the control diets. Specific antibody against S. iniae or E. tarda measured by ELISA did not reveal differences in the fish surviving the challenge. We conclude that the incorporation of these commercial yeast component products into the diet of juvenile Nile tilapia at these rates and for these feeding periods had no effect on growth, serum components, antibody responses, or survival following S. iniae or E. tarda infection.     

Dry whey meal as a protein source in practical diets for Nile tilapia,Oreochromis niloticus fingerlings

ABSTRACT

This study was conducted as a trial of using dry whey meal (DWM) as a substitute for fish meal (FM) in practical diets for Nile tilapia, Oreochromis niloticus. Triplicate fish groups were fed on five isonitrogenous (30.2%) and isolipidic (6.9%) diets. The control diet (D1) used FM as the sole protein source. In the other four diets (D2–D5), FM protein was substituted by 25, 50, 75, or 100% DWM. Fish (3.5 ± 0.1 g) were stocked at a rate of 20 fish per 100-L aquarium and fed one of the tested diets up to satiation twice daily for 12 weeks. Fish growth, feed utilization, protein efficiency ratio, apparent protein utilization, and energy utilization for fish fed DWM diets up to 75% FM (D2–D4) tended to be higher but were not statistically different than the control diet. No significant effect of diet was found in whole-body moisture, crude protein, and total ash contents. Whole-body lipid content in fish fed the 100% DWM (D5) diet was significantly higher than that for fish fed the control diet. The optimal replacement level of FM by DWM was estimated by second-order polynomial regression to be 62.5%.