Effects of canola meal on physiological and biochemical parameters in rainbow trout (Oncorhynchus mykiss)

Rainbow trout (initial body weight 4.16 ± 0.25 g) were fed diets [crude protein 420 g kg^?1; gross energy 18.7 MJ kg^?1 dry matter (DM); crude fat 110 g kg^?1] containing graded levels of either a canola meal (crude protein 350 g kg^?1 DM) supplemented with DL‐methionine as partial fish meal protein. A growth trial was conducted over 16 weeks at a water temperature of 12 ± 1 °C. At the end of the growth trial, in addition to body composition analyses, plasma tri‐iodothyronine (T₃) and thyroxine (T₄), cholesterol and liver fatty acid composition were measured. Replacement of fish meal with canola meal (100–570 g kg^?1 replacement) did not affect on growth performance. At 16th week, plasma cholesterol levels were reduced in fish fed all diets in comparison with 8th week. Plasma T₄ levels were significantly higher in the canola meal‐fed fish sampled after 16 weeks, but no significant differences in T₃ levels were obtained (P > 0.05). Proximate compositions were affected by dietary treatments. The liver fatty acid composition reflected that of the diet with a higher level of polyunsaturated (n‐6) fatty acids in fish fed diet canola meal and a higher content in n‐3/n‐6 ratio in fish fed diet without canola meal. These studies show that canola meal has potential to replace substantial levels of fish meal in diets for carnivorous fish without compromising performance.     

QTL for IHNV resistance and growth identified in a rainbow (Oncorhynchus mykiss) × Yellowstone cutthroat (Oncorhynchus clarki bouvieri) trout cross

Infectious hematopoietic necrosis virus (IHNV) is a major constraint to rainbow trout culture. Yellowstone cutthroat trout (Oncorhynchus clarki bouvieri) have greater resistance to this virus than do rainbow trout (O. mykiss), but the genetic mechanism of this resistance is not understood. We conducted a genome scan using a backcross of cutthroat trout into a rainbow trout background to estimate the number and locations of quantitative trait loci (QTL) associated with IHNV resistance and growth in trout. IHNV resistance was considered in terms of both survival (binary trait) and days to death (quantitative trait). The genetic map was scanned using interval mapping via two different approaches: one model considered survival alone and a second two-part model combined both survival and days to death. Three QTL were significantly (P ≤ 0.05) associated with virus resistance genome-wide, explaining 32.5% of the phenotypic variation. Cutthroat alleles at two of these QTL resulted in increased resistance to the pathogen, as expected. No growth QTL were detected in this cross. We suggest that these traits are genetically independent.     

Evaluation of fertilizing capacity of cryopreserved rainbow trout sperm

ABSTRACT:   Experimental insemination was performed using artificially produced low-motility sperm. A mathematical model was applied to the results of the insemination in order to clarify the relationship between sperm motility, the density of sperm and the fertilization rate of eggs. In the model, the probability of fertilization by individual spermatozoa was a function of sperm density in the insemination solution. The results showed that the probability of fertilization clearly decreased with increased sperm density, and the maximum possible fertilizing rate by increasing the sperm density was constrained by the proportion of motile sperm (% motility). The model was also applied to the results of insemination tests of cryopreserved sperm in order to evaluate the fertilizing capacity of cryopreserved sperm. It was proven that cryopreserved sperm needed a higher density to obtain the maximum fertilization rate compared with fresh sperm, and it was anticipated that the ratio of the motile inseminated cryopreserved sperm should be more than 5.0% to achieve an egg fertilization rate greater than 90%.     

不同倍性虹鳟卵黄蛋白原的组织定位

采用分步沉淀法提纯虹鳟卵黄蛋白,以同发育期二倍体虹鳟为对照,用免疫组化法对不同发育阶段的三倍体虹鳟性腺、肝脏、血液和肠道进行卵黄蛋白原(Vg)的细胞化学定位研究.试验结果表明:所纯化蛋白为与卵黄蛋白原具有相似免疫原性的卵黄脂磷蛋白,呈雌性特异性.性腺发育Ⅲ～Ⅴ期的二倍体雌性虹鳟,血液和肝细胞呈卵黄蛋白原阳性,Ⅳ～Ⅴ期卵巢呈卵黄蛋白原阳性,各发育期肠组织呈卵黄蛋白原阴性;性腺发育至Ⅰ～Ⅴ期的三倍体雌雄虹鳟及雄性二倍体虹鳟,其性腺、肝脏、血液和肠道组织呈卵黄蛋白原阴性.二倍体虹鳟外源性卵黄蛋白原在肝细胞内合成,经血液运送至卵巢,最终在卵母细胞中形成卵黄颗粒.由于三倍体雌性虹鳟卵巢发育受阻,无滤泡细胞分化,生殖细胞与体细胞的互作缺失,不能分泌足量的17β-E2以诱导Vg的合成,因而肝脏的Vg阴性并不能说明肝脏不具备合成Vg的能力,从卵黄发生的角度分析,卵黄蛋白原的缺乏不是导致三倍体虹鳟雌性不育的原因,而是其卵泡败育的结果.     

Soybean meal alters autochthonous microbial populations, microvilli morphology and compromises intestinal enterocyte integrity of rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout were fed either a diet containing fishmeal (FM) as the crude protein source or a diet containing 50% replacement with soybean meal (SBM) for 16 weeks. An enteritis-like effect was observed in the SBM group; villi, enterocytes and microvilli were noticeably damaged compared with the FM group. The posterior intestine microvilli of SBM-fed fish were significantly shorter and the anterior intestine microvilli significantly less dense than the FM-fed fish. Electron microscopy confirmed the presence of autochthonous bacterial populations associated with microvilli of both fish groups. Reduced density of microvilli consequently led to increased exposure of enterocyte tight junctions, which combined with necrotic enterocytes is likely to diminish the protective barrier of the intestinal epithelium. No significant differences in total viable counts of culturable microbial populations were found between the groups in any of the intestinal regions. A total of 1500 isolates were tentatively placed into groups or genera, according to standard methods. Subsequent partial 16S rRNA sequencing revealed species that have not been identified from the rainbow trout intestine previously. Compared with the FM group levels of Psychrobacter spp. and yeast were considerably higher in the SBM group; a reduction of Aeromonas spp. was also observed.     

Effects of the extensive culture system as finishing production strategy on biometric and chemical parameters in rainbow trout

The effect of finishing extensive farming period, to reduce fat content and manipulate the fatty acid profile of fish muscle, was evaluated in rainbow trout. Fish were stocked in an artificial lake, in which fish were fed only on naturally available nutrients with no supply of artificial feed, for different lengths of time from 0 to 120 days. No weight loss was noted during the whole finishing period while total length increased from 228±7 to 269±3 mm and the condition factor decreased from 1.41±0.04 to 0.89±0.02. The total fat content of the fillets decreased considerably from 4.7±0.6% at the beginning to 2.4±0.4% and 0.7±0.2% after 45 and 120 days respectively. Fillet fatty acid composition was affected by the time of stocking in the extensive farm. In contrast to the reduction in C18:1n‐9, C18:2n‐6, total monounsaturated fatty acid and total n‐6 percent values, an increase in the C20:5n‐3, C22:6n‐3, total polyunsaturated fatty acid and total n‐3 percent values was observed. It was shown that while other finishing strategies for salmonids have some disadvantages, the extensive culture system seems to be a potentially useful tool for increasing the general quality of the end product.     

二倍体和三倍体虹鳟精液低温保存寿命的研究

与二倍体(2n组)精原液做比较,以不同密度为组别,研究了4℃下两组(3n-HD和3n-LD)三倍体虹鳟(Oncorhynchus mykiss)精原液的保存时限。结果发现,随着保存时间的延长,精液中精子的活力和寿命逐渐降低。2n组、3n-HD组和3n-LD组虹鳟精原液中精子分别在保存5d、4.5d和12.5d后失去活力;在7d、15d和14.5 d后死亡,即三倍体虹鳟精原液4℃保存时间约为二倍体的2倍,说明低温短期保存三倍体虹鳟精原液可以取得较好效果。     

虹鳟IL-17多克隆抗体的制备及初步鉴定

本研究根据NCBI已发表序列设计引物,提取经植物血凝素刺激后的虹鳟Oncorhynchus mykiss头肾细胞总RNA,采用RT-PCR方法扩增IL-17成熟肽基因,测序结果表明所获得的序列与发表序列相一致。将该基因重组至原核表达载体pET32a中,并转化大肠杆菌Escherichia coli Rosetta,进行诱导表达。SDS-PAGE电泳结果表明:目的蛋白以包涵体形式表达,大小约为32k Da,重组蛋白经Ni-NTA系统纯化、复性后纯度达90%以上,以其免疫小鼠制备虹鳟IL-17多克隆抗体。ELISA结果显示其效价为1∶25 600,而间接免疫荧光结果显示所制备的多克隆抗体能够特异性地识别真核细胞中瞬时表达的虹鳟IL-17。本研究成功制备虹鳟IL-17多克隆抗体,为下一步IL-17在虹鳟黏膜免疫中的作用及其佐剂效应研究奠定基础。     

Dietary formic acid enhances apparent digestibility of minerals in rainbow trout, Oncorhynchus mykiss (Walbaum)

The effect of dietary formic acid on the availability of phosphorus (P) from a fishmeal-based diet by rainbow trout, Oncorhynchus mykiss (Walbaum), reared in brackish water (5–6 g L⁻¹) was determined. Diets containing a low amount of P (6.0 mg P g⁻¹ dry matter) were acidified with 0, 4 and 10 mL kg⁻¹ formic acid and fed to trout (average weight, 520 g) for 4 weeks. The measured pH of the diets were 6.3, 5.8 and 5.3, respectively. The intestinal solubility of P and the digestibility of P were measured by stripping the faecal matter from the fish. The apparent digestibility coefficient (ADC) of P significantly ( P < 0.05) increased from 69.5% to 75.0% of the basal diet in fish fed diets containing 10 mL kg⁻¹ formic acid. The solubility of P in the intestine was highly variable within each treatment, and the differences were not significant. The pH of intestinal content increased with the increase in dietary formic acid concentration. The ADC of magnesium and calcium also showed a significant ( P < 0.05) increase with the acidification of diet by formic acid.     

不同饲粮对虹鳟鱼卵中矿物质、维生素含量及胚胎发育的影响

试验研究了池塘人工培育下不同饲粮对虹鳟亲鱼所产2种颜色鱼卵中矿物质、维生素含量及对受精率、发眼率、破膜率的影响。结果表明:橘红色鱼卵Ca、Mg、Zn含量略高于淡黄色鱼卵(20.8—67.0—ng/kg),其他6种矿物质均为淡黄卵略高于橘红卵(0.063～46.0 mg/kg);2种颜色鱼卵的维生素含量有差别:淡黄色鱼卵的4种维生素的含量均大于橘红色鱼卵的含量,淡黄色鱼卵中维生素D含量显著大于橘红色鱼卵(P<0.01);2种颜色鱼卵的受精率、发眼率、破膜率差异不显著(P>0.05)。