Effects of pretreatment with microbial phytase on phosphorous utilization and growth performance of Nile tilapia (Oreochromis niloticus)

This study was to assess effects of the pretreatment in all‐plant based diets with microbial phytase on phosphorous utilization and growth performance of Nile tilapia (Oreochromis niloticus). Pretreatment trials were conducted using phytase at graded doses to determine the optimal dose of phytase. Available phosphorus (P) levels increased significantly with the increased doses of phytase and the dose of 1000 U kg^?1 was most efficient. Based on the pretreatment trials, plant based diets for Nile tilapia were formulated by pretreating with phytase at 1000 U kg^?1. Experimental diets were supplemented with graded levels of mono calcium phosphate (MCP) at 25, 18.75, 12.5, 6.25 and 0 g kg ^?1 diet. In addition, there were three controls: one phytase control, one inorganic P control and one pretreatment control. The results showed that diets pretreated with phytase gave better growth performance, feed conversion ratio and protein efficiency ratio of Nile tilapia compared with the phytase control diet and pretreatment control diet (P < 0.05). There were no significant differences in growth performance of Nile tilapia between the inorganic control diet and phytase pretreated diets supplemented with MCP at 25, 18.75 and 12.5 g kg^?1 (P > 0.05), which resulted in significantly better performance than those at 6.25 and 0 g kg^?1 (P < 0.05). Dietary interaction effects of phytase were observed for phosphorus retention efficiency and phosphorus load. Apparent digestibility coefficient of P (ADCp) was improved significantly by phytase pretreatment (P < 0.05). No significant difference was detected on ADC of crude protein among all experimental diets (P > 0.05).     

Physiological and Biochemical Determination of Rainbow Trout,Oncorhynchus mykiss,Semen Quality for Cryopreservation

In rainbow trout, Oncorhynchus mykiss, parameters to determine semen fitness for cryopreservation and quality control of cryopreserved semen were investigated. The following parameters can be used to evaluate semen fitness for cryopreservation as they are statistically significant (P < 0.01) correlated to the post-thaw fertilization rate: motility rate of fresh semen (y = 4.996x - 0.0958x² + 0.0006x³ - 5 1.7363); sperm velocity of fresh semen (y = 6.741x - 0.036x² - 268.37); seminal plasma osmolality (y = 0.539x - 125.59); seminal plasma pH (y = -82.768x + 728.133); seminal plasma triglyceride levels (y = 0.069x + 29.863); seminal plasma ß-D-glucuronidase activity (y = -1.112x + 0.0058x² + 82.229); seminal [lasma lactate dehydrogenase activity (y = -0.096x + 0.00006x² + 583.80); spermatozoan acid phosphatase activity (y = -132.51x + 126.38x² + 66.48); spermatozoan adenylate kinase activity (y = 3.474x + 4.925). Quality of deep-frozen semen can be evaluated by motility parameters (P < 0.01): frozen/thawed semen motility rate and post-thaw fertilization rate: y = 1.943x + 28.002; sperm velocity and post-thaw fertilization rate: y = 0.8812x - 0.0059x² + 24.9686.     

Evaluation and Nutrient Quality of Detoxified Jackbean Seeds,Cooked in Distilled Water or Trona Solution,as a Substitute for Soybean Meal in Nile Tilapia,Oreochromis niloticus,Diets

ABSTRACT

Jackbean, Canavalia ensiformis, seeds were detoxified using eight different wet thermal detoxification methods in either distilled water (5% w/v) or trona (NaCO₂.NaHCO₃.2H₂O) solution. The effects of the detoxification methods on hemagglutinating, antitryptic and urease activities, and on the digestibility coefficient of crude protein and gross energy in the jackbean seeds were investigated. Cracked jackbean seeds cooked (100°C) in trona solution for one hour proved to be more effective as a method of inactivating the antinutritional factors in jackbean seeds. Dried detoxified jackbean seed meals were incorporated in dry practical diets providing 20% or 30% of total protein and fed to Nile tilapia, Oreochromis niloticus, fingerlings (mean weight, 6.7 g) twice daily to apparent satiation for 70 days. Mortality was low and no abnormal fish behavior was noted even when the detoxified jackbean meals provided 30% of total protein in diet. Growth and feed utilization indices, were similar (P > 0.05) between the diet in which cracked jackbean seeds cooked in water provided 20% of total protein and in diets in which cracked jackbean seeds cooked in trona solution provided 20% or 30% of total protein. Carcass composition and hepatosomatic index showed no definite trend among O. niloticus fingerlings fed the experimental diets.     

Re-evaluation of total sulphur amino acid requirement and determination of replacement value of cystine for methionine in semi-purified diets of juvenile Nile tilapia, Oreochromis niloticus

Two feeding experiments were conducted to re-evaluate the total sulphur amino acid (TSAA; methionine and cystine) requirement and determine the replacement value of cystine for methionine of juvenile Nile tilapia ( Oreochromis niloticus ). Semi-purified diets used in both experiments contained 3510 kcal gross energy and 280 g of protein per kilogram diet from casein, gelatin and crystalline amino acids. The basal diet of the first experiment contained 3.1 g methionine and 0.4 g cystine per kilogram. l -methionine was added to the seven remaining diets at 1.0 g kg⁻¹ increment to produce methionine levels ranging from 3.1 to 10.1 g kg⁻¹ diet. Each diet was fed to four replicate groups of juvenile Nile tilapia (1.28 g mean weight) in a recirculation system for 8 weeks. Broken-line regression analysis of weight gain data indicated that the TSAA requirement of juvenile Nile tilapia was 8.5 g kg⁻¹ of the diet or 30.4 g kg⁻¹ of dietary protein. In the second experiment, TSAA level was set at 95% of the requirement value determined in the first experiment. Seven diets were made with different ratios of l -methionine and l -cystine (20 : 80, 30 : 70, 40 : 60, 50 : 50, 60 : 40, 70 : 30 and 80 : 20, based on an equimolar sulphur basis). Each diet was also fed to four replicate groups of juvenile Nile tilapia (4.14 g mean weight) in a recirculation system for 8 weeks. Regression analysis of weight gain data using broken-line model indicated that cystine (on a molar sulphur basis) could replace up to 49% of methionine requirement in semi-purified diets for juvenile Nile tilapia.     

Spontaneous diploidization of the maternal chromosome set in Nile tilapia (Oreochromis niloticus L.) eggs

Spontaneous diploidization of the maternal chromosome set (SDM) in Oreochromis niloticus is described here for the first time. The SDM phenomenon was observed in progeny of only one XY neofemale out of 11 such neofemales from which eggs had been fertilized with UV‐irradiated sperm: this treatment produced only the expected haploid embryos from the other females. SDM progeny were produced from three different batches of eggs from this female. No significant differences in survival at different stages of embryonic development were observed between the SDM and the diploid control group. The maternal inheritance of SDM progeny was verified using multilocus DNA fingerprinting. Only diploid karyotypes were observed in these fish and diploid control groups. In both the SDM and control groups, sex ratios were significantly skewed towards males (93.3% and 65.0% males respectively). The actual mechanism of SDM in eggs from this particular female remains unknown.     

Time related alterations in digestibility and faecal characteristics as affected by dietary composition in the Nile tilapia (Oreochromis niloticus L.)

After being shifted to a new diet, time related alterations in digestibility, faecal waste production and faeces recovery in Nile tilapia were assessed in relation with dietary ingredient composition. Four experimental diets were formulated according to a 2 by 2 factorial design: two starch inclusion levels (20 or 40%) and two maize starch types (native versus gelatinized). After introduction to the experimental diets, faeces were collected weekly from Week 2 till 6 using settling tanks. Digestibility of ash, organic matter and dry matter increased with time (P < 0.001). For organic and dry matter the time related alteration in digestibility were different between both starch types (P < 0.05). All faecal waste characteristics altered with time (P < 0.01). The incline in faeces recovery with time was affected by starch type (P < 0.01); being the largest at the 40% gelatinized maize diet. In conclusion, the minimal length of the adaptation period in digestibility studies for obtaining unbiased digestibility estimates is dependent on diet composition, in this study with Nile tilapia 4 weeks for diets with gelatinized starch and 6 weeks with native starch.     

Fisheries by-catch and processing waste meals as ingredients in diets for Nile tilapia, Oreochromis niloticus

Digestibility, feeding and growth studies were conducted with Nile tilapia using diets containing fishery by‐catch and processing waste meals. Three meals manufactured from sorted fisheries by‐catch (MBM, from mixed benthic species, SPM, from small pelagic species, CAM, from mixed catfish species) one from tuna cannery waste (TCW) and one commercial anchovy meal (COM) were tested. By‐catch and processing waste meals had lower protein, lower lipid and higher ash contents than anchovy meal. The meals were all highly digestible and no significant differences (P≥0.05) were observed between apparent protein digestibility measurements. Five feeds, containing fish meal as the major protein ingredient, were formulated and fed to triplicate groups of 30 juvenile tilapias for 9 weeks. Survival, weight gain, specific growth rate (SGR), feed conversion ratio, protein efficiency ratio, phosphorus retention and whole body proximate composition were compared. Weight gain and SGRs were similar for each treatment group and compared favourably with the results obtained from juvenile tilapia elsewhere. Growth was the highest for CAM (P≤0.05), which contained both the highest essential amino acid levels and the highest ash content. Phosphorus retention was significantly lower in fish fed with high ash meals, MBM, CAM and TCW (P≤0.05) than in fish fed with the lower ash meals COM and SPM. Overall, the fisheries by‐catch and processing waste meals evaluated in this study are suitable protein ingredients for juvenile tilapia feeds.     

Cortisol Response of Nile Tilapia,Oreochromis niloticus (L.), to Temperature Changes

Abstract

Nile tilapia, Oreochromis niloticus (L.), is a commercially-valuable fish species with high nutritional value. As a result of the intensive aquaculture of this species, handling, transport, and environmental changes that causes stress on these fish are unavoidable. The objective of this study was to determine the effects of gradual and acute temperature changes on juvenile tilapia. No significant difference (P > 0.05) was found among serum cortisol levels in juvenile tilapia when the water temperature was gradually increased from 27°C to 32°C, or 40°C, and maintained for 1 hour, although the levels were five times pretreat-ment levels. When tilapia acclimated to 27°C were subjected to 18°, 27°, 30°, 32°, 34°, 36°, 38°, or 40°C water for 10 minutes in a water bath, followed by a recovery period of 10 minutes at 27°C in the original aquaria, no significant difference (P > 0.05) in cortisol levels was observed among treatments except for significantly elevated levels at 38°C and 40°C. When tilapia acclimated to 27°C were subjected to the same temperature exposures but given a recovery period of 60 minutes at 27°C in the original aquaria, there was no significant (P > 0.05) increase in cortisol levels in tilapia among treatments from 18° to 36°C; but there was a significant (P > 0.05) increase between values from those treatments at 38° and 40°C. Acute temperature changes initiated the cortisol response as early as 10 minutes in fish following exposure to 38°C or 40°C and resulted in significant increases in the 38°C and 40°C treatments following 1 hour of recovery at 27°C. These results have implications for the management of tilapia during bacterial challenge, vaccination, and handling and transport during aquacultural activities.     

尼罗罗非鱼淀粉酶性质的初步研究

研究了尼罗罗非鱼(Oreochromis niloticus)肝脏淀粉酶的基本性质及金属离子对该鱼淀粉酶活性的影响。结果表明:罗非鱼淀粉酶活力的最适pH是6.5,最适底物浓度是2%。研究金属离子对该鱼淀粉酶活力影响,一价金属离子K+、Li+、Na+对酶活力影响较小;二价金属离子Cu2+对酶活力具有抑制作用,Zn2+对酶活力无明显影响;三价金属离子Al3+对酶活力具有抑制作用,但效果不是很强烈,Fe3+对酶活力具有明显的激活作用;碱土金属Mg2+、Ca2+对淀粉酶活性有激活作用,而Ba2+对酶具有抑制作用;重金属离子Cd2+、Pb2+有明显的抑制作用。     

Effects of Dietary β-Hydroxy-β-Methylbutyrate on Growth and Survival of Nile Tilapia,Oreochromis niloticus,Vaccinated Against Streptococcus iniae

Abstract

β-hydroxy-b-methylbutyrate (HMB), a leucine catabolite, has been shown to cause increased disease resistance and growth in animal production. A vaccine produced from formalin killed bacteria and concentrated extracellular products of the ARS-98-60 Streptococcus iniae isolate has been used for the prevention of streptococcal disease in Nile tilapia, Oreochromis niloticus. In the present study, the effects of feeding HMB were determined in tilapia vaccinated by intraperitoneal (IP) injection of the S. iniaevaccine or unvaccinated (controls). Nile tilapia were fed diets containing either 0, 12.5, 25, or 50 mg HMB/kg diet for 14 days. The mean daily growth rate and feed efficiency showed no significant (P> 0.05) differences between the treatment groups. Dietary HMB supplementation did not enhance antibody production in unvacci-nated Nile tilapia following challenge. Dietary HMB supplementation did not enhance the survival of vaccinated Nile tilapia following challenge injection with 1 X10⁸ CFU of S. iniae.     

高温诱导对尼罗罗非鱼性别分化及生长的影响

采用自雌鱼口腔中取出受精卵人工孵化的方法建立了尼罗罗非鱼家系20个。受精后9d,将各家系鱼苗分成两份,分别在28℃(对照组)和36℃(高温诱导组)下养殖,诱导处理12d后,统计高温诱导期成活率并将实验鱼转入自然水温下培育,养殖60～120d后对各实验组的雄性率及生长情况进行测定。结果表明,高温处理对仔鱼的成活率没有影响,但能显著提高某些家系的雄性率,如F27(诱导组88%,对照组49%)、F37(诱导组90%,对照组53%)和F41(诱导组91%,对照组53%);高温处理后多数家系的雄性率略有升高,但不显著;有两个家系高温诱导组雄性率低于对照组,分别是F8(诱导组43%,对照组46%)和F35(诱导组29%,对照组38%),这显示了尼罗罗非鱼性别分化受遗传和环境因子共同作用。生长情况测定结果表明,高温诱导对发育初期罗非鱼的生长没有显著影响。本研究结果说明,高温处理能显著提高某些家系的雄性率,高温诱导和家系选择有望成为高雄性率尼罗罗非鱼新品种培育的新的技术手段。     

三种生态环境下尼罗罗非鱼免疫因子的比较

随机选取海水池塘(盐度28)和淡水池塘养殖及水库天然增殖的3种不同生态环境下、体质量为200~500g的尼罗罗非鱼为研究材料,采用生物化学方法测定3种生态环境尼罗罗非鱼血清中碱性磷酸酶、酸性磷酸酶、过氧化物酶、谷胱甘肽还原酶的活性和蛋白质量浓度。结果显示,3种生态环境中罗非鱼血清中碱性磷酸酶和酸性磷酸酶活力差异不显著(P0.05),其余各组间罗非鱼血清中过氧化物酶和谷胱甘肽还原酶的活性及血清蛋白质量浓度差异显著(P0.05),酶活力和蛋白质量浓度依次为:水库海水池塘淡水池塘。试验结果表明,水库天然增殖的尼罗罗非鱼具有较高的磷酸酶活性。     

Effect of dietary probiotic Biogen® supplementation as a growth promoter on growth performance and feed utilization of Nile tilapia Oreochromis niloticus (L.)

Probiotic microbial feed supplements are gaining wide acceptance in livestock production, and may be applicable to aquaculture production systems. The present study was conducted to examine probiotic treatment in the fingerling diet of Nile tilapia Oreochromis niloticus (L.). A total of 240 of Nile tilapia fingerlings (weight ranged from 22.96 to 26.40 g) were divided into five experimental groups. The experiment was conducted for 120 days. Experimental diets were identical in all, except for the variation in probiotic levels. A probiotic (Biogen^®) was used at 0% (diet 1), 0.5% (diet 2), 1.5% (diet 3), 2.0% (diet 4) and 2.5% (diet 5) inclusion rates in the experimental diets. The growth performance and nutrient utilization of Nile tilapia including weight gain, specific growth rate, protein efficiency ratio, protein productive value and energy retention were significantly (P≤0.01) higher in the treatment receiving probiotic (Biogen^®) than the control diet. No differences were observed for moisture, ash and protein content (P≤0.01) among the experimental diets. The lowest gross energy and lipid contents were recorded for fish fed the diet containing 0.5% Biogen^® (P≤0.01). The production performance and subsequent cost–benefit analyses clearly indicated that the diets containing probiotic biogen recorded the highest net return and the lowest total cost compared with the control diet.     

低温胁迫对尼罗罗非鱼水通道蛋白(Aquaporin 1，AQP1)基因(AQP1)表达的影响

以经过连续多代抗寒选育获得的尼罗罗非鱼(Oreochromis niloticus)耐寒品系为实验材料, 克隆了尼罗罗非鱼水通道蛋白基因(AQP1)的cDNA序列, 并采用实时荧光定量PCR分析方法探讨了低温胁迫对罗非鱼AQP1基因表达水平的影响。序列分析结果显示, 尼罗罗非鱼AQP1 cDNA长度为1 098 bp, 包含36 bp的5′端非翻译区序列、783 bp开放阅读框(ORF)和279 bp的3′非翻译区序列, 编码261个氨基酸。氨基酸序列比对表明, 各物种间AQP1序列的同源性较高(96%~59%)。聚类分析表明, 尼罗罗非鱼首先与同属鲈形目丽鱼科的斑马拟丽鱼(Maylandia zebra)聚在一起, 再与其他硬骨鱼类聚为一类。实时荧光定量PCR 结果表明: 在水温从30℃逐渐降到10℃过程中, 肌肉和肝组织AQP1基因的表达量从20℃开始均逐渐下调。其中肌肉组织中, 与30℃时的表达量相比, 耐寒品系在20℃时表达量大幅度下调, 当水温为15℃时, 其表达量下调幅度为22.16倍。当水温降到10℃时, 其表达量下调了107.73倍。而对照组AQP1基因的表达量在15℃时, 下调幅度为5.38倍。当水温达到10℃时, 其表达量下调幅度为11.30倍。结果分析显示, 耐寒品系与对照组AQP1基因在低温条件下的表达量存在显著性差异(P<0.05)。而在肝组织中, 当水温降到15℃时, 耐寒品系和对照组的表达量下调幅度分别为3.38倍、1.42倍。水温降到10℃时, 其表达量上调幅度分别为18.85倍、9.01倍。结果分析表明, 低温条件下耐寒品系与对照组AQP1基因在肝组织中的表达差异不显著(P>0.05)。结果表明, AQP1表达对温度敏感, 耐寒品系较高的耐寒能力与其在低温条件下的肌肉组织大幅上调表达有关。由此可见, AQP1基因在罗非鱼低温适应过程中发挥重要作用, 是潜在的研究罗非鱼耐寒机制的候选基因之一, 本研究为进一步研究罗非鱼的耐寒分子机制和开展鱼类抗逆育种提供参考。

     

Use of ideal protein concept for precision formulation of amino acid levels in fish-meal-free diets for juvenile Nile tilapia (Oreochromis niloticus L.)

This study was undertaken in a closed system with Nile tilapia (Oreochromis niloticus) to examine the effects of total replacement of fish meal (FM) by soybean meal. Nile tilapia fingerlings with an average weight of 5.34±0.08 g were hand‐fed one of the five isoenergetic (≈13.5 MJ digestible energy kg^?1) and isoproteic (≈31% of digestible protein) experimental diets to satiation, six times a day during 85 days in eight replicate fibreglass tanks (six fish per tank). The control diet containing FM was substituted by soybean meal, with and without essential amino acids (lysine, methionine and threonine) or dicalcium phosphate supplementation. The supplemental amino acids were added at levels to simulate the reference amino acid profile of Nile tilapia carcass protein, based on the ideal protein concept. The results showed that soybean meal diet supplemented only with dicalcium phosphate was inferior to the control diet with FM and soybean meal diets supplemented with dicalcium phosphate and essential amino acids. Multiple essential amino acids and dicalcium phosphate incorporation in soybean meal diets was associated with performance, whole‐body composition and carcass yield equal to that of the fish fed with the control diet containing FM. These data suggest that a diet with all plant protein source, supplemented with essential amino acids, based on tissue amino acid profile, can totally replace FM in a diet for Nile tilapia, without adverse effects on the growth performance, carcass yield and composition.     

Production of heterozygous and homozygous clones in Nile tilapia

Production of heterozygous and homozygous clones of Nile tilapia (Oreochromis niloticus L.) was successfully carried out. An outbred clonal line was produced by cross-breeding between a viable mitotic gynogenetic female and male (recessive mutation in a sex determining gene). At the same time, an inbred clonal line was produced by gynogenetic reproduction (retention of 2nd polar body) using optimal pressure/heat shock treatments. The heterozygous and homozygous nature of the two types of clonal lines were checked and identified at the adenosine deaminase ADA* marker locus. A model for the large scale production of similar clones is presented. The possible implications of such clonal lines in genetic research and in the culture of Oreochromis spp. are discussed. © Rapid Science Ltd. 1998     

Use of microsatellite loci and AFLP markers to verify gynogenesis and clonal lines in Nile tilapia Oreochromis niloticus L.

To develop an effective system for parentage analysis in gynogenetic and clonal progeny of Nile tilapia, Oreochromis niloticus L., polymorphic microsatellite loci and amplified fragment length polymorphisms (AFLPs) were investigated in several gynogenetic families and clonal lines. Six microsatellite loci were screened in two meiotic gynogenetic families to look for loci with high gene–centromere recombination rates, which can be used to discriminate meiotic from mitotic gynogenetics. Microsatellite loci UNH189 and UNH211 showed 96.7% and 92.0% heterozygosity, respectively, in these families, while other loci showed lower recombination frequencies. Scoring both UNH189 and UNH211 would give a very low probability of an individual meiotic gynogenetic being homozygous for both loci. Multiplex polymerase chain reaction of microsatellite loci was used to verify parentage in four families of mitotic gynogenetics and five fully inbred clonal lines. The genotype of each clonal line should serve as a unique identifier. Twelve AFLP primers were also investigated and 26 diagnostic AFLP bands were identified to follow inheritance in mitotic gynogenetic individuals. Amplified fragment length polymorphisms were found to be effective for this purpose but microsatellites were more appropriate since they are co‐dominant, while AFLPs are dominant markers. A multiplex of the microsatellite loci used in this study would be useful for general parental assignment as well as for the analysis of the products of chromosome set manipulations.     

Comparative digestion efficiencies in conventional, genetically improved and genetically male Nile tilapia, Oreochromis niloticus (L.)

Apparent digestibility of dry matter, protein, lipid and energy of a fishmeal‐based feed (41% crude protein, 9% crude lipid and 19 kJ g⁻¹ gross energy) was compared in genetically improved farmed tilapia (GIFT), genetically male Nile tilapia (GMNT) and conventional Nile tilapia (CNT) (Oreochromis niloticus). The experimental fish were reared individually under standardized conditions in a recirculation system at 27±0.1°C for 10 weeks. Titanium dioxide (TiO₂) was used as a marker. Faeces of individual fish were collected daily by siphoning and stored at −18°C before analysis. No significant differences (P<0.05) were observed in the digestibility coefficients of feed dry matter (78.2±3.4%, 77.7±4.4% and 76.4±3.7%), protein (87.9±3.0%, 88.4±2.8% and 88.0±3.3%), lipid (90.0±2.5%, 91.0±2.1% and 89.4±3.0%) and energy (90.4±1.9%, 90.7±2.0% and 89.4±2.3%) in GIFT, GMNT and CNT respectively. At the end of the experiment, there were no significant differences (P<0.05) in average percentage growth (82.2±7.2, 87.3±7.7 and 74.7±4.1 respectively for GIFT, GMNT and CNT), growth rates or feed utilization efficiencies between the three tilapia groups. We conclude that the higher growth claimed for improved GIFT and GMNT as compared with CNT, if ever existing, cannot be attributed to higher nutrients or energy digestibility.