Nutritional,physiological and haematological responses in rainbow trout (Oncorhynchus mykiss) juveniles fed detoxified Jatropha curcas kernel meal

Jatropha curcas is a multipurpose and drought‐resistant tree, widespread throughout tropics and subtropics. Its seeds are rich in oil and protein. It is being promoted as a biofuel plant. Jatropha kernel meal obtained after oil extraction is an excellent source of protein [580–660 g kg^?1 crude protein (CP)]. However, the presence of toxic and antinutritional constituents restricts its use in fish feed. Jatropha kernel meal was detoxified. Using Oncorhynchus mykiss juveniles, a 12‐week experiment was conducted to evaluate the nutritional quality of detoxified Jatropha kernel meal (DJKM). Oncorhynchus mykiss juveniles (36, av. wt. 4.2 ± 0.4 g) were distributed into three groups with 12 replicates and fed isonitrogenous (CP 450 g kg^?1) diets: control [fish meal (FM)–based protein], J₅₀ and J_62.5 (50% and 62.5% of fishmeal protein replaced by DJKM). Growth performance, apparent lipid conversion, hepatosomatic index, nutrients and energy digestibilities; efficiency of digestible nutrients and energy; and glucose and creatinine levels in blood were statistically similar (P > 0.05) for control and J₅₀ but were higher (P < 0.05) than for J_62.5. Opposite trend was observed for phosphorus, sodium, albumin and total protein in blood. Feed conversion ratio, protein efficiency ratio, protein productive value, energy retention, gastro somatic index, muscle lipid peroxide; and red and white blood cells, haematocrit, haemoglobin, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration; alkaline phosphatase and alanine transaminases activity, globulin, total bilirubin, blood urea nitrogen, calcium and potassium in blood and lysozyme activity in serum did not differ significantly among the groups. Intestinal enzyme activities; and plasma and muscle cholesterol levels for control were higher (P < 0.05) than for DJKM groups, while opposite trend was observed for relative intestinal length (mm g^?1). Conclusively, performance of J₅₀ group was similar to control group and, thus, is recommended as the optimal diet for rainbow trout.     

Effect of dietary bovine lactoferrin on growth,haematology and non‐specific immune response in rainbow trout (Oncorhynchus mykiss)

The aim of this study was to investigate the effect of dietary lactoferrin (Lf) on growth, haematology and non‐specific immune response of rainbow trout Oncorhynchus mykiss (Walbaum). Fish were fed an experimental diet containing 0 (as control), 50, 100, 200 and 400 mg Lf kg^?1 diet twice daily for 8 weeks and sampled at 2, 4, 6 (immune function and growth), and 8 weeks (immune function, haematology and growth). Statistical analyses revealed no significant effects of dietary Lf on growth performance (specific growth rate, weight gain, feed conversion ratio, feed intake and condition factor) or haematological parameters (red and white blood cell count, haemoglobin, haematocrit, serum iron and total iron binding capacity [TIBC]). Among the serum non‐specific immune parameters, lysozyme activity increased significantly in fish fed 100, 200, or 400 mg Lf kg^?1 feed for 8 weeks, whereas haemolytic complement activity increased in fish fed 100 and 400 mg Lf kg^?1 diet after 6 weeks. The antiprotease activity increased in groups fed 100, 200 or 400 mg Lf kg^?1 diet after 8 weeks. However, no significant effect was observed on serum peroxidase level. It can be concluded that feeding of rainbow trout on the diet supplemented with 100 mg kg^?1 or higher for 8 weeks enhances the non‐specific immune response.     

Antioxidant and immune defences of rainbow trout (Oncorhynchus mykiss) offered plant oils differing in fatty acid profiles from early stages

The antioxidant defence and immune response of rainbow trout (Oncorhynchus mykiss) that had received plant oils, rich in either n‐3 polyunsaturated fatty acids (PUFA; linseed oil) or n‐6 PUFA (safflower oil) was evaluated upon antigen exposure. The fish employed in this study had been offered the diets for 18 months from the first feeding. Rainbow trout from each group were injected intraperitoneally with formalin‐killed bacteria (Aeromonas hydrophila) or were sham‐injected and observations were made 24 h later. Though the fish fed safflower oil seemed to be under relatively greater oxidative stress, the antioxidant defences (superoxide dismutase, catalase, glutathione peroxidase) were as effective as in those fed linseed oil. The humoral (alternate complement activity and lysozyme activity) and cellular (phagocytic activity and lymphocyte proliferation) immune responses were not significantly affected by the oil offered. With the exception of reactive oxygen production that was significantly greater in the linseed oil fed fish, both groups did not differ greatly in their immune responses after antigen exposure. Thus, fish fed safflower oil that was deficient in n‐3 PUFA was able to sustain most of the critical responses similar to those fed linseed oil suggesting that plant oils of both fatty acid categories were effective for this fresh water fish.     

Covariation between diploid and triploid progenies from common breeders in rainbow trout, Oncorhynchus mykiss (Walbaum)

The covariation between diploid and triploid progenies from common breeders was investigated in various progeny-testing experiments where either dams or sires were sampled from rainbow trout, Oncorhynchus mykiss (Walbaum), stocks. Triploidiza tion was found to frequently reduce the performance in the traits studied: body length and weight, growth, coefficient of condition and pyloric caeca number. Triploidization also generated significant interactions with the parental breeding value. These interactions were caused in part by the familial variance not being the same in triploids as in diploids, but also by actual ranking differences between diploid and triploid familial performances. However, the effect of these interactions was minor as compared with the amount of variation common to both ploidy levels (genetic correlations averaged 0.7–0.9). Therefore, selection of diploid breeders appeared efficient enough for improving triploid progeny, unless family selection methods including triploid progeny testing were preferred for other reasons. Lastly, it was observed that variances from maternal origin tended to be larger in triploids, whereas variances from paternal origin tended to be smaller, as compared with diploids. This point was discussed referring to the genetic make-up of triploids and in the scope of dams and sires for selective breeding.     

Interaction between diet and genetic aptitude for weight and growth in juvenile rainbow trout, Oncorhynchus mykiss (Walbaum)

Samples of rainbow trout, Oncorhynchus mykiss (Walbaum) alevins from 17 independent families (full‐sib‐groups) were raised from the start of feeding until the age of 18 weeks post‐hatching with three diets (C, G and S) differing in protein content (fish soluble protein concentrate: 84% in C, 54% in G and 44% in S) and carbohydrates (none in C, 30% glucose in G and 40% crude corn starch in S). Fish were fed to near satiation, and their body weight and growth were measured. Diet effect was highly significant (G< C < S), as well as the familial effect. The major part of the familial variance (80–90%) was common to the three diets. However, a minor part of the familial variance was observed to be diet dependent (family × diet interaction), and was found to result mainly from relative performances with carbohydrates (G and S diets) vs. pure protein (C diet). These results indicate that genetic improvement of growth should suffer little impairment from possible changes in future feed formulations.     

Intestinal absorption and tissue distribution of glucose and isoleucine in rainbow trout (Oncorhynchus mykiss)

The intestinal absorption and tissue distribution of D(U¹⁴C)-glucose (¹⁴C-glc), 3-O-methyl-D(U¹⁴C)-glucose (¹⁴C-OMG) and L(U¹⁴C)-isoleucine (¹⁴C-ile) were studied in rainbow trout, Oncorhynchus mykiss, using a forced-feeding technique. The appearance of radioactivity in faeces, blood, liver, white muscle and brain was monitored over 48 h. The recoveries of radioactivity 48 h postfeeding in the fish tissues studied were 36, 57 and 48% for ¹⁴C-glc, ¹⁴C-ile and ¹⁴C-OMG, respectively. White muscle and liver contained most radioactivity on the whole tissue basis. Concentrations of ¹⁴C-glc and its derivatives in the liver and the brain exceeded those of ¹⁴C-ile and its derivatives, but the reverse was found in the white muscle. These differences may reflect differences in the metabolism of glucose and isoleucine. All tissues studied showed some differences in the accumulation of ¹⁴C originating from ¹⁴C-OMG and ¹⁴C-glc. As ¹⁴C-OMG is a nonmetabolizable glucose analogue, these differences may be the result of glucose metabolism at tissue level.     

Dietary microbial phytase supplementation and the utilization of phosphorus, trace minerals and protein by rainbow trout [Oncorhynchus mykiss (Walbaum)] fed soybean meal-based diets

Effects of thermal and enzymatic treatments of soybean meal on apparent absorption of total phosphorus, phytate phosphorus, nitrogen (protein), ash, calcium, magnesium, copper, iron, manganese, strontium and zinc were examined using rainbow trout, Oncorhynchus mykiss (Walbaum), as the test species. Absorption of the test nutrients was estimated using yttrium as an inert non-absorbable indicator. Thermal treatments (microwaving, dry roasting, steam heating, cooking) had no measurable effect on the apparent absorption of phosphorus and other minerals. Phytase supplementation increased the apparent absorption of phosphorus, nitrogen (protein), ash, calcium, magnesium, copper, iron, strontium and zinc in low-ash diets containing soybean meal, but had little effect in high-ash diets containing both soybean and fish meal. In low-ash diets, the apparent absorption of phosphorus increased in accord with the level of phytase added to the diet, from 27% (no phytase added) up to 90% (phytase added, 4000 units kg⁻¹ diet) or 93% (predigested with phytase, 200 units kg⁻¹ soybean meal). In high-ash diets, dietary acidification with citric acid decreased the effect of phytase, whereas in low-ash diets, acidification markedly increased the effect of the enzyme. Excretion of phosphorus in the faeces of fish fed a low-ash diet containing phytase-treated soybean meal was 0.32 g per kg diet consumed, a 95%−98% reduction compared with phosphorus excretion by fish consuming commercial trout feeds.     

Dietary essential amino acids and heat increment in rainbow trout (Oncorhynchus mykiss)

Oxygen consumption attributable to apparent heat increment (AHI) was measured in relation to varying essential amino acid proportions (EAA) infused into rainbow trout,Oncorhynchus mykiss (250–450 g), induced to swim at 1 BL s^–1. Five diets, mimicking EAA concentrations in trout whole body protein, deficient in the branched chain amino acids (isoleucine, leucine and valine), containing unbalanced proportions of EAAs and supplying lysine in excessive and limiting proportions, were tested. Following infusion of the experimental diets, a significant increase in oxygen consumption was observed. Changes in plasma EAAs following infusion paralleled the time course of AHI (i.e., oxygen consumption). AHI represented the equivalent of 15–32% of the gross energy intake depending on dietary EAA composition. Diets supplying EAAs similar to trout whole body protein and limiting in lysine produced the lowest AHI values, indicating efficient utilization of dietary amino acids. Higher AHI values were associated with diets deficient in the branched chain amino acids and diets supplying lysine in excess. Duration of elevated metabolism was independent of both dietary composition and energy intake. Different proportions of EAAs in the diet can increase the energy expended as AHI. In an attempt to reduce the energy liberated as AHI, attention must be paid to the quality, quantity and balance of dietary EAAs.     

Effect of dietary inclusion of lysozyme on growth performance and plasma biochemical parameters of rainbow trout (Oncorhynchus mykiss)

This study was conducted to evaluate the effect of dietary lysozyme supplementation on growth performance and plasma biochemical parameters of rainbow trout (Oncorhynchus mykiss). Graded levels of lysozyme [0 (control), 150, 300, 450 and 600 mg kg^?1 diet] were fed to fish (initial average weight 7.76 g) for 10 weeks. Dietary lysozyme supplementation regardless of inclusion level generally improved the growth, feed utilization and nutrient retention, but significant differences were mainly observed between the 450 or 600 mg kg^?1 lysozyme group and the control group. Lysozyme supplementation generally decreased the plasma aspartate aminotransferase and alanine aminotransferase (ALT) activities, but significant difference was only observed in the ALT activity between the 450 mg kg^?1 lysozyme group and the control group. In contrast, lysozyme supplementation generally increased the plasma alkaline phosphatase and lysozyme activities, but significant difference was only observed in the lysozyme activity between the 600 mg kg^?1 lysozyme group and the control group. Lysozyme supplementation generally increased the plasma antioxidant activities of enzymes (including superoxide dismutase, glutathione peroxidase and catalase) and total antioxidant capacity, but decreased the plasma malondialdehyde level. These results suggest that dietary 450–600 mg kg^?1 lysozyme inclusion may improve the growth and non‐specific immune response of rainbow trout.     

The use of AquaMats® to enhance growth and improve fin condition among raceway cultured rainbow trout Oncorhynchus mykiss (Walbaum)

AquaMats® are a type of artificial seaweed designed to provide structure in ponds used for fish culture and as a substrate for the growth of aquatic plants and invertebrates which in turn are a source of nutrition to cultured species. In two separate tests AquaMats® were placed into raceways used to rear rainbow trout Oncorhynchus mykiss (Walbaum) to evaluate their effect on fish growth and fin condition. In the first test, the AquaMats® were placed perpendicular to the raceway length similar to a baffle design. One treatment consisted of AquaMats® that were cleaned on a regular basis, and the other treatment consisted of AquaMats® that were not cleaned throughout the test. By the end of the test no differences were found between treatments with respect to final fish weight, specific growth rate, or feed conversion ratio. The use of AquaMats® did not improve fin condition, in fact several fins measured were significantly better among control fish. In the second test AquaMats® were placed on the raceway bottom parallel to their length and to the water flow. AquaMats® were also hung from the side of the raceway to provide cover. At the conclusion of this test no differences were found between treatments with respect to final fish weight, specific growth rate, or feed conversion ratio. The placement of AquaMats® did have a transitory impact on fin condition. Mid‐way through the test, treatment fish generally exhibited longer fins compared with the controls. However, by the end of the test, these differences were no longer detectable. The results from both tests indicate that fish were not provided with additional nutrition to the extent it improved growth. However, the use of AquaMats® did make a significant, albeit transitory, impact on fin condition.     

Partial or total replacement of fishmeal by solvent-extracted cottonseed meal in diets for juvenile rainbow trout (Oncorhynchus mykiss)

The effect of solvent‐extracted cottonseed meal (SCSM) as a partial or total replacement of fishmeal was studied in juvenile rainbow trout (Oncorhynchus mykiss). Six experimental diets SCSM0, SCSM25, SCSM50, SCSM75, SCSM75A and SCSMT, containing a gradient of SCSM 0, 152, 305, 465, 460 and 610 g kg^?1 to replace 0, 112.5, 225, 337.5, 337.5 and 450 g kg^?1 fishmeal protein were fed to triplicate groups (initial body weight of 39.2 ± 0.1 g) for 8 weeks. The diet SCSM75A was supplemented with lysine and methionine, to be similar to SCSM0 for juvenile rainbow trout. Faeces were colleted after 4 weeks of normal feeding for apparent digestibility coefficients (ADC) of dry matter, crude protein and gross energy determination. Total replacement of fishmeal adversely affected growth performance. Fish fed with diet SCSMT had significantly (P < 0.05) lower weight gain, specific growth ratio, feed conversion efficiency (FCE) and protein efficiency ratio than fish fed with other diets. The FCE of SCSM75 and SCSM75A were significantly lower (P < 0.05) than those of fish fed with SCSM0 diets. The ADC of the dry matter of SCSM75 and SCSMT were significantly lower than the SCSM0 diet, and the ADC of crude protein and the energy of SCSMT were the lowest (P < 0.05). The ADC of threonine, proline, alanine, valine, isoleucine, leucine, lysine and methionine of fish fed with diet SCSMT were lower. Lysine and methionine supplement positively affected the ADC of SCS75A diet. There were no significant differences in the fish body composition. It is shown that SCSM can be utilized in the juvenile rainbow trout diet up to 305 g kg^?1, to replace about 50% of fishmeal protein in this experiment.     

A test of a feed budget model for rainbow trout, Oncorhynchus mykiss (Walbaum)

A feed budget model, which enables farmers to adapt a general model to local conditions, was tested on rainbow trout, Oncorhynchus mykiss (Walbaum). The model has two major components; component 1 is used to calculate the energy required for one unit of biomass gain (g or kg) and component 2 estimates the daily growth increment (g or kg). Both components can be adjusted in relation to prevailing environmental conditions and, together, they express the daily energetic requirement of the specific species or strain. Fish fed using the model as a guide had a high rate of growth and good feed conversion. The model accurately predicted growth over the experiment, and is presented as a means of calculating the daily feed budget in fish farming. Growth of fish fed using self-feeders is presented for comparison with the results obtained using the model.     

Utilization of (3S,3'S)-astaxanthin acyl esters in pigmentation of rainbow trout (Oncorhynchus mykiss)

The deposition of natural, optically active, astaxanthin fatty acid esters in rainbow trout ( Oncorhynchus mykiss ) was studied. Mono-esterified and di-esterified (3 S ,3' S ) astaxanthin were purified from the green microalga Haematococcus pluvialis and incorporated into extruded diets and compared with diets containing synthetic racemic astaxanthin (Carophyll Pink) and a total carotenoid extract from the alga. All sources of astaxanthin achieved >4 mg kg⁻¹ in the white muscle after 6 weeks feeding. No significant difference ( P  > 0.05) between the deposition of astaxanthin or total carotenoid for the different diets was observed. Other xanthophylls, namely lutein, zeaxanthin and idoxanthin were found in the white muscle of rainbow trout fed all diets and together accounted for 10–14% of total carotenoid. Astaxanthin was deposited in the white muscle in the stereochemical form administered in the diet, i.e. racemic astaxanthin for Carophyll Pink and ˜100% (3 S ,3' S )-astaxanthin for the algal sources. In contrast, epimerization of (3 S ,3' S ) astaxanthin from the alga was observed for the astaxanthin esters deposited in the skin of rainbow trout, with a ratio close to 1.0:2.0:1.0 (3 S ,3' S :3 R ,3' S :3 R ,3' R ).     

Myosin expression levels in trout muscle: a new method for monitoring specific growth rates for rainbow trout Oncorhynchus mykiss (Walbaum) on varied planes of nutrition

Fish growth is manifested by a number of measurable physical changes. We have developed a sensitive method for monitoring the growth rate of fish fed at four different planes of nutrition. This technique consists in measuring expression levels of myosin RNA isolated from the muscle of experimental animals. Using PCR (polymerase chain reaction) primers, and a fluorescence-labelled single-stranded DNA probe that hybridizes specifically to a region within the myosin mRNA of rainbow trout Oncorhynchus mykiss (Walbaum), we were able to detect differences in the relative level of myosin expression between groups of fish. This method also allows for the determination of absolute expression levels when reactions are performed with standards consisting of known levels of in vitro-transcribed myosin RNA. With the proper equipment, this novel procedure can be performed rapidly on large numbers of individuals, and with the procurement of non-invasive muscle biopsies the same experimental animal could theoretically be sampled multiple times throughout the course of the study. This new method could be used to measure differences in muscle synthesis in fish associated with various nutrient intake levels, environmental parameters, life-history stages and health status.     

The effectiveness of adenoviral vectors to deliver and express genes in rainbow trout,Oncorhynchus mykiss (Walbaum)

The efficacy of adenoviral vectors for gene delivery into fish cells, both in vitro and in vivo, was evaluated. Vectors utilized were of human adenovirus serotype 5 (Ad), which are commonly used in human clinical trials, but have not been assessed for gene delivery to fish. Because nothing is known about Ad receptors in fish, both an Ad (Ad5Luc1) with natural tropism for the coxsackie and adenovirus receptor (CAR), as well as an infectivity enhanced Ad (Ad5LucRGD) were included within this study. Gene expression was detected in cell lines using either vector. The levels seen with Ad5LucRGD were much higher than for Ad5Luc1 in most lines except CHSE-214. Transduction of CHSE-214 cells with Ad5Luc1 could be blocked with an excess of a competitive inhibitor, suggesting that these cells possess a CAR homologue thatmediates attachment of Ad, similar to that seen in mammalian cells. In vivo gene delivery was attempted by several methods, with significant expression seen only via intramuscular injection, although infection efficiency was low. Thus it was observed that several teleost cell lines are capable of being infected and one cell line expressed a human serotype adenoviral receptor homologue that aids in Ad infection. Additionally, in vivo studies indicated that muscle tissue of rainbow trout could be infected with Ad vectors, suggesting an alternative gene delivery strategy for this animal.     

Effects of dietary visceral protein hydrolysate of rainbow trout on growth performance,carcass composition,digestibility and antioxidant enzyme in juvenile Oncorhynchus mykiss

The purpose of this study was to investigate the effects of dietary visceral protein hydrolysate (VPH) of rainbow trout on performance of juvenile Oncorhynchus mykiss. The antioxidant property of VPH was 85% ± 1.6%. Four diets were prepared including VPH free as the control and three diets with 5, 10 and 20 g/kg of VPH inclusions. Juvenile rainbow trouts (n = 252, 9.74 ± 0.22 g) were randomly distributed to experimental tanks. Final results showed the maximum amounts of weight gain and SGR and the lowest FCR value in fish fed with 10 g/kg VPH (p < .05). The highest carcass protein and fat contents were observed in 10 g/kg VPH treatment and the control diet, respectively (p < .05). The largest fat and dry matter digestibility were recorded in fish fed by 5 g/kg VPH (p < .05), while the control and 10 g/kg VPH diets showed the highest levels of protein digestibility (p < .05). Dietary inclusion of VPH improved SOD enzyme activity (p < .05). The results indicate that a diet containing 10 g/kg of VPH inclusion can improve growth performance and some other examined factors in juvenile O. mykiss.     

Reproductive performance of female rainbow trout Oncorhynchus mykiss (Walbaum) kept under water temperatures and photoperiods of 13° and 51°N latitude

At Doi Inthanon Fisheries Research Unit (DIFRU), Thailand (13°N), rainbow trout were exposed to natural (13°N) and artificial (51°N) photoperiods, and natural water (NW) temperatures and cooled water (CW) 8 months before first spawning. In group I (51°N, CW), water temperatures of 18°C were never reached. In group II (51°N, NW) and group III (13°N, NW), the mean water temperatures in May exceeded 20°C, and 19°C in June and July. Eggs from 94% of all females in group I were obtained before January. This percentage diminished to 84% and 68% in groups II and III. The weight of the spawners and the size of the eggs were significantly lower in group III than in the other groups. No significant differences were observed for egg number per kg body weight of spawners between the groups. The mean fertilization rate of eggs was the highest, with 71%, in group I, and the lowest, with 50%, in group II. For hatching rates, on average 27%, 24% and 30% in groups I, II and III, respectively, differences were not significant. In group III, 37% of all batches reached fertilization rates above 80% and 16% of egg batches showed hatching rates of more than 60%.     

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。