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。     

Streptococcus agalactiae Vaccination and Infection Stress in Nile Tilapia,Oreochromis niloticus

Abstract

The stress response following intraperitoneal (IP) injection of a non-adjuvant Streptococcus agalactiae vaccine in cultured warmwater Nile tilapia, Oreochromis niloticus, has not been investigated. Further, little or no information is available on stress following S. agalactiae infection and what effect, if any, vaccination has on susceptibility to infection. The objective of this study was to develop preliminary information on the associations between vaccination, stress, and infection. Blood glucose levels were used to evaluate stress in the fish at different time intervals following vaccination and challenge with S. agalactiae. Blood glucose levels were measured in vaccinates and controls at 0, 2, 6, 24 hours, and 28 days post-immunization (0 hours pre-challenge), and at 2,6, 24,48,72, and 312 hours following challenge with 1.5 × 10⁴ colony forming units (CFU) of S. agalactiae/fish. Significant increases in blood glucose levels were observed only in association with the injection of the vaccine and at 2 hours after injection. After S. agalactiae challenge, both controls and vaccinates had significantly (P < 0.05) higher blood glucose values at 2, 24,48, and 72 hours than at 0 hours. However, blood glucose levels in vaccinates were significantly (P < 0.05) lower than the controls at 24, 48, and 312 hours. Blood glucose levels and mortality of the infected controls were significantly correlated (r² = 0.9236, P = 0.0134). The cumulative mortality of the vaccinates and controls was 10% and 60% after 13 days post-challenge, respectively. The relative percent survival (RPS) was 83.4. Our results indicate that the vaccine was efficacious against S. agalactiae and induced short-term stress in tilapia. These preliminary results also suggested, for the first time, that vaccination may significantly reduce the infection stress associated with S. agalactiae infection in tilapia.     

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。     

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.     

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.     

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.     

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.     

Passive immunization of tilapia, Oreochromis niloticus (L.), with anti-Streptococcus iniae whole sera

Passive immunization of tilapia, Oreochromis niloticus, was conducted to determine whether anti- Streptococcus iniae whole sera (ASI), heat inactivated anti- S . iniae whole sera (HIASI) and normal whole sera (NWS) were protective when intraperitoneally (i.p.) injected into tilapia. The ASI was produced in tilapia actively immunized (challenged) with virulent S. iniae by i.p. injection. An antibody response against S. iniae was demonstrated by enzyme linked immunosorbent assay (ELISA) and 18% of the immunized fish died because of the S. iniae infection. The actively immunized tilapia demonstrated a secondary antibody response and immunity to S. iniae after challenge with S. iniae by i.p. injection. Survival was 100% in the actively immunized fish. The NWS was obtained from tilapia free of ASI antibody and susceptible to S. iniae infection (40% mortality). In two separate experiments, significantly higher mortality was noted in tilapia passively immunized with NWS (33 and 53%) and phosphate buffered saline (PBS) (30 and 60%), in comparison with mortalities of 0 and 10% or 3.3 and 6.7% in the fish passively immunized with ASI or HIASI 14 days after S . iniae infection by i.p. injection ( P  = 0.0003 and 0.0023). Results suggest that immunity provided by ASI and HIASI was because of antibody against S. iniae . Inactivation of complement in the HIASI treatment further suggests that ASI antibody plays a primary role in immunity against S. iniae infection.     

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。     

Characterization and comparison of variations in reproductive performance of Chitralada strain Nile tilapia, Oreochromis niloticus (L.)

In an attempt to better understand and characterize the variability in the female reproduction of individual Thai‐Chitralada strains of Nile tilapia, Oreochromis niloticus (L.), tagged mature females (n=68) from a single population were monitored in a hapa‐in‐pond system over a 12‐month experimental period. Spawn female⁻¹, eggs female⁻¹ day⁻¹, eggs kg female⁻¹ day⁻¹, day spawn⁻¹ and weight (g) at first spawning of individual females were determined from the regular weekly sampling of weight (g) and eggs per spawn. For analysis, the females were grouped into two, high spawning (HSF) and low spawning frequency (LSF) classes based on their spawning frequency (SF) record. Moreover, nested under these two classes were two groups each based on growth rate, i.e., high frequency – large size (HL), high frequency – small size (HS), low frequency – large size (LL) and low frequency – small size (LS). There was no difference in eggs spawn⁻¹ among all females. The HSF group produced 68% and 361% more eggs female⁻¹ day⁻¹ than the population mean and LSF group respectively. Eggs female⁻¹ day⁻¹ and spawn female⁻¹ remained high in the HSF group and low in the LSF group throughout the 12‐month experimental period. This suggests that individual female spawning activity is consistent within a population in a common environment. The inter‐spawn interval increased with age in all four groups, and day spawn⁻¹ was shorter by 130% in HSF females compared with day spawn⁻¹ in LSF. The HSF group also spawned more successively (≥3), while the LSF group of females had fewer successive spawns (≤2). Body weight (g) had no influence on the number of eggs produced. The study indicates that separating frequently spawning females could be used as an important strategy to improve commercial seed production of Nile tilapia.     

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.     

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.     

Simulation of Nile tilapia (Oreochromis niloticus niloticus L.) culture in ponds, through individual-based modelling, using a population dynamic approach

A simulation model for the production of the Nile tilapia (Oreochromis niloticus niloticus L.) for mixed‐ or mono‐sex culture and for poly‐culture with the African catfish (Clarias gariepinus Burchell 1822) or African snakehead (Parachanna obscura Günther 1861) is presented. The model is based on an exponential decay model used in population dynamics and follows an individual‐based approach. The model consists of a tilapia sub‐model representing population dynamics, growth and recruitment and a predator sub‐model representing the predation process and population dynamics and growth of the used predator. The model was calibrated with data on mixed‐ or mono‐sex culture of Nile tilapia and for poly‐culture with the African catfish or African snakehead obtained in Congo Brazzaville and validated with similar data from the Philippines, Thailand and the Ivory Coast. The model visualized major underlying processes in tilapia farming and aspects for further improvement of the model; growth is one of the most sensitive input parameters and should be quantitatively related to feeding level and feed quality; length at first maturity and quantification of the recruitment of Nile tilapia has a relatively large influence and recruitment should be related to the length of the females; prey–predator relations are too coarse and should be more fine‐tuned with the relation between prey size and predator size. Incorporation of these features would provide the basis of a model that can serve as a predictive and decision‐making support tool.     

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.     

Influence of Dietary Lipid Sources on the Growth Performance,Immune Response and Resistance of Nile Tilapia,Oreochromis niloticus,to Streptococcus iniae Challenge

ABSTRACT

This experiment was conducted to determine the effect of dietary lipid sources on growth performance, body proximate composition, hematology, immune response and resistance of Nile tilapia, Oreochromis niloticus, to Streptococcus iniae infection. Six isocaloric (3.2 kcal/g) and isonitrogenous (34% crude protein) semi-purified diets were supplemented with 7% of various sources of lipid, namely, corn oil (CO), beef tallow (BT), menhaden fish oil (FO), linseed oil (LO), and equal combinations of FO+CO+BT or LO+CO+BT. Diets were fed to tilapia in quadruplicate aquaria to apparent satiation, twice daily for 12 weeks. Fish fed the BT-diet exhibited significantly lowest weight gain, diet intake, feed and protein efficiency ratios, apparent protein utilization, and survival. Whole-body protein and ash were significantly (P < 0.05) lowest and highest, respectively, for fish fed the beef tallow-diet, but the values of these parameters did not differ among fish fed other diets. No significant differences (P > 0.05) were found among hematological values, except for fish fed the FO-diet which had abnormally high red and white blood cell counts. Serum protein concentration, lysozyme activity, and natural hemolytic complement activity were significantly (P < 0.05) reduced in fish fed the BT-diet. The values of these parameters did not differ among fish fed other diets. Post-challenge antibody titer was not influenced by dietary lipid sources. Cumulative mortality 15 days post-challenge with S. iniae was significantly lower (P < 0.05) for fish fed the BT diet compared with those fed FO or FO+CO+BT diets. No significant differences were observed in fish fed other dietary lipid sources.     

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.     

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).     

Evaluation of visible implant elastomer tags for pathogenesis research in Nile tilapia (Oreochromis niloticus)

Two different colours (red and green) of visible implant elastomer (VIE) were used in Nile tilapia (Oreochromis niloticus). The visibility, location and retention of the VIE tags was investigated and any adverse effects on fish survival and growth determined. The use of VIE tags for monitoring individual fish during a bacterial challenge with either Streptococcus agalactiae or S. iniae was also studied. The results showed that VIE treated fish were lighter but not shorter than the non‐tagged control fish and that tagging caused no mortality. The retention of tags was better at the base of pectoral fin followed by the nasal area, lower abdomen, upper abdomen and branchiostegal rays inside the operculum. During the bacterial challenge experiment individual animals could be easily identified using the VIE tags. In this preliminary study, VIE tagging appears suitable for Nile tilapia research, as with other fish species, and could be a novel method to identify individual animals during microbial pathogenesis studies.     

木薯粉在罗非鱼饵料中的适口性研究

选择健康的吉富罗非鱼为试验鱼,比较不同水平木薯粉之间的诱食效果以及木薯粉与豆粕、次粉的适口性,评价木薯粉在罗非鱼饵料中的应用效果。结果表明,(1)40%、30%木薯粉组的诱食活性显著高于10%、20%木薯粉组(p0.01),40%与30%木薯粉之间没有显著差异(p0.05)。(2)罗非鱼对木薯粉与豆粕、次粉的适口性依次为豆粕木薯次粉。