吉富品系尼罗罗非鱼（GIFT）摄食节律初探

用投喂添加色素的膨化颗粒饲料和观测排便相结合的方法,研究吉富品系尼罗罗非鱼(Genetic Improvement of Farmed Tilapias;GIFT)的摄食节律.结果发现,6个家系的吉富品系尼罗罗非鱼摄食具有明显的节律性,家系之间的摄食周期存在非常显著差异(P＜0.0001);t检验发现,各家系内摄食周期和排便周期的差异均不显著(P≥0.1097).回归分析发现,摄食周期对生长无显著的影响(P=0.8988),但与摄食量之间存在显著的线性关系(R2=0.9679,P=0.0004).对摄食节律聚类分析,6个家系可分为(1、2、6)、(3、4)、5共3个家系类别.该项研究能为GIFT的养殖投饵和进一步选育提供理论依据,为鱼类摄食节律研究提供方法上的参考.     

Molecular cloning and gene expression of the riboflavin-binding protein in the Nile tilapia, Oreochromis niloticus

Tilapia riboflavin-binding protein (RfBP) cDNA was isolated by library screening. Even though it shows only 31～35% similarity to the reported RfBPs, tilapia RfBP is conserved in amino acid residues that are known to be essential for its protein function. The exclusive expression of RfBP in the brain-pituitary-gonadal axis, as revealed by both RT-PCR and Northern blot, suggested a possible role for RfBP in fish reproduction.     

Buprofezin toxication implicates health hazards in Nile tilapia (Oreochromis niloticus)

The objective of this study was to investigate the toxic effects of buprofezin insecticide on Nile tilapia (Oreochromis niloticus). The fish were exposed to buprofezin at 100 mg/L for 28 days. Compared to control, activity of serum transferases and levels of urea and creatinine showed significant increases. Oxidative stress was recorded manifested by elevated levels of malondialdehyde (MDA), reduced concentrations of reduced glutathione (GSH) and inhibition of activities of superoxide dismutase (SOD) and catalase (CAT) in liver and kidney. Examination of peripheral RBCs revealed elevated frequency of micronucleated cell. Interleukin 1 beta (IL‐1β) gene was upregulated in liver, muscle and brain, while that of cyclooxygenase 2 (COX‐2) gene increased in liver and muscle, but not in brain. Histopathological alterations were recorded in liver, kidneys, brain, gills, pancreas, spleen, intestine, muscle and ovaries. The immunohistochemical detection of caspase‐3 in the liver revealed no differences between treated and control groups; however, the expression of inducible nitric oxide synthase (iNOS) was demonstrated in hepatocytes and hepatopancreas in buprofezin‐treated group compared to control. It has been concluded that the tissue damage induced by buprofezin in Nile tilapia is mediated by oxidative stress and inflammatory response but not by apoptosis.     

罗非鱼谷胱甘肽过氧化物酶1基因的克隆与分析

采用cDNA末端快速扩增（rapid amplification of cDNA ends,RACE）技术,克隆了罗非鱼（Oreochromis niloticus）谷胱甘肽过氧化物酶1（glutathione pemxidasel,GPx1）基因完整的编码序列（complete coding sequence,CDS）。GPx1基因全长984bp,5’uTR56bp,CDS576bp,3’UTR352bp,PolyA20bp;第791—885位碱基（位于3’UTR）形成1个硒半胱氨酸插入序列（selenocysteine insertion sequence,SECIS）,协助174～176位密码子TGA（UGA）编码1个硒半胱氨酸（Sec）。GPx1包含191个氨基酸,分子量21．8kDa,等电点8．04,无信号肽和潜在的N．糖基化位点。蛋白结构分析表明该基因编码蛋白为非跨膜蛋白。序列比对显示,GPx1单体具有Sec、Trp、Gln和Asn构成的催化四联体。罗非鱼GPx1与其他脊椎动物GPx1相比较,核苷酸序列相似性为43．2％-58．2％,氨基酸序列相似性为58．1％～80．6％。进化分析显示,处在分类学上不同纲的脊椎动物GPx1分别占据了不同分支。利用Swiss-Model预测了罗非鱼GPx1的3D结构,序列分析显示,GPx1可以形成1个同源四聚体。     

Spermatogenesis in teleost: insights from the Nile tilapia (Oreochromis niloticus) model

The morphometric study of spermatogenic cysts in sexually mature tilapias, during the evolution of spermatogenesis, showed a dramatic increase in both number of germ cells and cyst volume. However, the opposite trend was observed for germ cell size. Nevertheless, the number of Sertoli cells increased gradually up to leptotene/zygotene cysts, stabilizing thereafter. Based on the number of spermatids supported by each Sertoli cell and compared to mammals, Sertoli cell efficiency in tilapias is remarkably high. Sertoli cell proliferation was frequently observed, mainly in spermatogonial cysts, and probably is the major factor related to the testis growth and the increase in sperm production that normally occurs in adult tilapias. The combined duration of spermatocytes (5 days) and spermiogenic (5–6 days) phases of spermatogenesis in fish kept at 25 °C was 10–11 days. Mainly due to acceleration in meiosis, these two phases lasted a total of 6 days in tilapias kept at 30 °C, in the opposite way, at 20 °C spermatogenesis was arrested at pachytene spermatocytes. To our knowledge, this is the most comprehensive investigation performed up to date on testis morphometry and function in adult tilapias.     

Glycerol inclusion in the diet of Nile tilapia (Oreochromis niloticus) juveniles

The present study evaluated the effects of inclusion of glycerol in the diet of Nile tilapia juveniles on growth performance, biochemical changes in blood, and carcass composition. We used 300 Nile tilapia juveniles with an average initial weight of 29.15 ± 8.40 g and 11.55 ± 0.87 cm in length, distributed in 20 fiberglass tanks with a capacity of 250 L. The experiments followed a completely randomized design with five treatments and four replications during 79 days. The animals were fed diets containing four concentrations of glycerol (25 g kg^?1, 50 g kg^?1, 75 g kg^?1 and 100 g kg^?1) and a control diet without glycerol. HDL was the only biochemical parameter, that showed statistically different (P < 0.05) results; it was higher in the groups fed with 0 and 75 g kg^?1 glycerol compared to the other groups. No significant difference was observed in the results from the carcass composition of tilapia juvenile fed with the different glycerol levels, except for lipids (P < 0.05), which showed the highest values in fish fed with 50 g kg^?1 glycerol and the lowest in fish fed with 100 g kg^?1. Glycerol can be used in fish diets as an energy supplement without causing damage to growth performance or to the biochemical and carcass composition of Nile Tilapia juveniles.     

Sex reversal in Nile tilapia (Oreochromis niloticus Linnaeus) by androgen immersion

Two experiments were conducted to investigate the efficacy of three androgens applied through immersion treatments on the sex ratio of Nile tilapia (Oreochromis niloticus L.) fry. In experiment 1, 14 days post‐hatching (DPH) larvae were exposed to a single immersion treatment in 17α‐methyltestosterone (MT), 17α‐methyldihydrotestosterone (MDHT) or 17α‐ethynyltestosterone (ET) at 200, 600 and 1800 μg L^?1 over 4 h (130 larvae per treatment). In experiment 2, Nile tilapia larvae were exposed to the higher androgen concentration (1800 μg L^?1) applied as either a single immersion (14 DPH) or double immersion (10 and 14 DPH) over 4 h (125 larvae per treatment). Change in sex proportion within each experiment as well as between experiments was analysed by the chi‐square test. In experiment 1, MT, MDHT and ET were equally effective in significantly increasing the proportion of males when applied at 1800 μg L^?1 (86.0%, 90.0% and 86.7% respectively). At 200 μg L^?1 none of the androgens altered sex ratio. At 600 μg L^?1, only MDHT slightly, but significantly skewed the sex ratio towards males (73.0%). In experiment 2, a single immersion treatment at 14 DPH (1800 μg L^?1) significantly increased the proportion of males, but at this time the response was significantly hormone dependent (MDHT, 100.0%; MT, 91.6%; ET, 76.9%). When compared with a single immersion, two‐immersion treatments significantly increased the proportion of males in the MT‐treated group (from 91.6% to 98.3%), decreased the proportion of males in the MDHT group (from 100.0% to 93.4%) and had no significant effect the ET‐treated group (change from 76.9% to 82.5%). The overall comparison of the sex ratio among same treatments from different experiments (a single immersion in 1800 μg L^?1) was not significantly different.     

Isolation and molecular characterization of streptococcal species recovered from clinical infections in farmed Nile tilapia (Oreochromis niloticus) in the Philippines

Streptococcosis cause severe losses for global tilapia farming, especially in developing countries. The aim of this study was to identify and characterize streptococci recovered from Nile tilapia farmed in the Philippines. Moribund and apparently healthy fish were sampled from grow-out cages, ponds and hatcheries. Clinical signs observed included exophthalmia, eye opacity, ascites, lethargy, erratic swimming and haemorrhages. Results showed that both Streptococcus iniae and Streptococcus agalactiae were associated with disease in these sites. Consistent with global reports, including those from South-East Asia, S. agalactiae was more widespread than S. iniae. Molecular serotyping of the S. agalactiae isolates identified the serotype Ia and serotype Ib. Histopathological findings were meningitis, meningoencephalitis and septicaemia. Identical virulence profiles were found for all strains of S. iniae, while S. agalactiae strains were separated into virulence profile I and profile II. All strains were susceptible to the tested antibiotics and resistant to oxolinic acid. Only S. agalactiae serotype Ib showed resistance to sulphamethoxazole–trimethoprim. This is the first study from the Philippines to characterize the streptococci involved in disease outbreaks in tilapia aquaculture. Outputs from this study will promote the development of efficacious disease control strategies in tilapia farming for the Philippines and South-East Asia.     

Molecular and histopathological characterization of Photobacterium damselae in naturally and experimentally infected Nile tilapia (Oreochromis niloticus)

Mass mortality has occurred among cultured Nile tilapia, Oreochromis niloticus, on fish farms in Manzala, Dakahlia province, Egypt, in the summer season, 2019. Moribund fish were reported with deep ulcers, septicaemic lesions and sampled for bacterial isolation. In this study, most isolates were subjected to bacteriological examination, antibiotic sensitivity test, 16S rRNA gene sequencing and histopathological examination. Following isolate identification, intraperitoneal challenge of Nile tilapia with a bacterial suspension 2 × 10⁶ CFU/ml was performed. Samples from liver, spleen and kidney were collected for histological and biochemical analysis. The results showed a high similarity (99%) to Photobacterium damselae strains using phylogenetic analysis of 16S rRNA. P. damselae exhibited resistance to amoxicillin and erythromycin, as well it was highly sensitive to chloramphenicol and doxycycline. Moreover, haemorrhage, oedema, hemosiderosis and melanomacrophage activation in the liver and head kidney of infected fish were detected by light and electron microscopy. Also, significant higher levels of CAT and SOD in the spleen and head kidney, as well as the serum levels of NO were observed in experimentally challenged O. niloticus, compared to the control fish. Our data identified P. damselae for the first time from infected Nile tilapia, describing its sensitivity to a variety of antibiotics, histopathological alterations and oxidative stress impact, and it could be useful indicators for understanding P. damselae pathogenesis, which might provide a preventive efficacy for P. damselae.     

Improving spawning synchrony in the Nile tilapia, Oreochromis niloticus (L.)

Abstract. The development stage and number of seed (eggs and fry) removed from individual incubating Oreochromis niloticus (L.) broodfish at 10-day interharvest intervals (IHI) was compared with the harvest of free-swimming fry from undisturbed fish held in a tank within the same system. Spawning synchrony and productivity were compared for (1) total exchange of females for ‘conditioned’ females after each harvest, (2) exchange of only spawned females, and (3) return of females back into the spawning tank. Improved synchrony and seed production were positively related to removal of eggs and fry from incubating females and could have important implications for broodstock management in commercial hatcheries. Allowing fish to remain undisturbed in clear water tanks may lead to inhibition of spawning activity related to the formation of hierarchies. Individual weight loss by females during incubation was correlated with stage of seed harvested and thus with timing of spawning during the interharvest interval.     

Molecular characterization and increased expression of the Nile tilapia, Oreochromis niloticus (L.), T-cell receptor beta chain in response to Streptococcus agalactiae infection

The complete cDNA sequence of the Nile tilapia T-cell receptor (TCR) β chain was cloned using 5' RACE. The full-length, 1263-bp cDNA contained a 942-bp open reading frame (ORF) encoding a 314-amino-acid protein. Sequence analyses revealed that the Nile tilapia TCR β chain contains four conserved cysteine residues involved in the formation of disulphide bridges and a conserved amino acid motif believed to be important for assembly and signalling of the TCR αβ/CD3 complex, both of which are normally found in the TCR β chain of other vertebrates. As detected using semi-quantitative and quantitative RT-PCR, the highest expression level of TCR β was detected in the thymus. Interestingly, Streptococcus agalactiae significantly induced the up-regulation of the TCR β chain, and the strongest up-regulation was detected in the brain and peripheral blood leucocytes (PBLs). In in vitro experiments, concanavalin A and Aeromonas hydrophila were found to significantly increase the expression of the TCR β chain in PBLs after 48 h (P < 0.01) and 72 h (P < 0.05), respectively. Furthermore, real-time PCR analysis showed that intraperitoneal injection (IP) of 10(7) cfu mL(-1) of S. agalactiae could induce TCR β expression that was greater than the expression observed following administration of 10(9) cfu mL(-1). The presence of the TCR β chain in fish detected in this study suggests the presence of T-cell populations that have been found in higher vertebrates, which may play a crucial functional role in the response to fish pathogens.     

Cascading effects of introduced Nile perch (Lates niloticus) on the foraging ecology of Nile tilapia (Oreochromis niloticus)

Abstract –  Nile tilapia ( Oreochromis niloticus L.) is the dominant of the introduced tilapiines in many East African lakes and has flourished in the presence of introduced Nile perch ( Lates niloticus L.). We explored the hypothesis that O. niloticus exhibits increased omnivory in response to a decline in abundance of haplochromine cichlids. First, we quantified variation in habitat use and diet of O. niloticus in Lake Nabugabo, Uganda. Second, we compared the diet of O. niloticus in lakes with (Nabugabo, Victoria) and without (Mburo, Wamala, Nyamusingiri, Kyasanduka) introduced Nile perch. In Lake Nabugabo, a higher level of phytoplanktivory was observed in small juveniles than in larger fish and in wetland ecotone areas where haplochromines were most abundant. An omnivorous diet dominated by detritus and invertebrates was recorded for O. niloticus in lakes Nabugabo and Victoria, while a predominantly herbivorous diet was characteristic of O. niloticus in lakes without Nile perch. Availability of a broad food base in lakes where inshore insectivores have been reduced may explain the increased omnivory recorded in lakes Nabugabo and Victoria.     

Reproductive performance in female strains of Nile tilapia, Oreochromis niloticus

The Nile tilapia is among the most cultivated tropical fish in the world, yet little is known about the variations in reproductive characteristics among different strains. There is an interest in evaluating the reproductive performance of females of three tilapia strains. All strains were represented in equal proportion (n = 225), being named as Supreme (SUP), Premium Aquabel (PA), and Chitralada (CHI). These fish were cultivated for 7 weeks in the Aquabel Pisciculture Station (Brazil), from January to March 2011. The PA was the largest strain, with the highest values in all the morphometric parameters we analyzed (weight, total length, standard length and body depth). Among the evaluated reproductive parameters, the PA strain differed significantly from the other two in number and weight of eggs/female, volume of eggs/spawn and number of eggs/kg of fish. The percentage of non-spawning females in the SUP and CHI strains differed significantly from one another, while results for PA showed intermediate values and no significance compared to the other strains (P > 0.05). Among the reproductive parameters analyzed, the percentage of spawned females/week, more than one spawns during 7 weeks, and hatching rate showed no significant differences between strains (P > 0.05). There was a significant correlation (r = 0.69) between water temperature and percentage of spawned females/week. When evaluating the interval between two consecutive spawns, we observed that periods of 21 days were the most frequent in all strains. In conclusion, the strain that showed the best results in all morphometric and in most reproductive parameters we evaluated was the PA.     

美国品系尼罗罗非鱼的选育及其效果分析

采用闭锁群体继代选育的方法(Closed population continuing selection and breeding method)对美国品系尼罗罗非鱼(Oreochromisniloticus)进行提纯选育,并利用选育获得的尼罗罗非鱼亲本作为母本,奥利亚罗非鱼(Oreochromis aureus)作为父本进行杂交生产子一代(简称"美奥"罗非鱼),再对子一代开展网箱养殖对比实验,以养殖实验的结果来评价选育尼罗罗非鱼的效果。结果表明:(1)实验组美国品系尼罗罗非鱼雌性后备亲本留种率P2世代分别比第P0、P1世代提高了103.16%和35.92%,P1世代比P0世代提高了49.47%,远高于同期未经选育的美国品系尼罗罗非鱼;(2)实验组"美奥"罗非鱼杂交组合的雌性亲本产苗量高达21.5万尾,高于对照组的21.2万尾;实验组产苗前期、高峰期和后期后代雄性率分别为96.27%、97.01%和95.70%,高于对照组的94.37%、95.65%和94.18%,但均呈现先升高后降低的变化趋势;(3)经过选育的"美奥"罗非鱼平均体长、平均体质量、绝对增重率、绝对增长率、特定生长率和生长指标均显著高于对照组(P0.05);600g以上的罗非鱼所占的比例由对照组的51.2%增加到64.6%,增幅达到26.2%;头部比例有变小的趋势,头长/体长在0.30以下的比例由对照组的19.0%上升到32.07%,增幅达68.8%;实验组"美奥"罗非鱼的雄性率、单产及肥满度等主要经济指标均比对照组显著提高(P0.05)。以上结果表明,"美奥"罗非鱼的选育效果良好,可为"尼奥"罗非鱼的苗种生产提供亲本保障。     

Immuomodulatory effect of dietary turmeric supplementation on Nile tilapia (Oreochromis niloticus)

This study aimed to assess the immunomodulatory effect of dietary turmeric (TUM) in Oreochromis niloticus juveniles. A total of 240 Oreochromis niloticus fingerlings were divided into four equal groups (n = 60), three replicates (n = 20) for each: control and TUM‐treated groups at 2 (T1), 4 (T2) and 8 (T3) g kg^?1 diets for 8 weeks. Body weight gain, leucocytes counts, plasma levels of interleukin 2 (IL‐2), interleukin 4 (IL‐4), lymphocyte proliferation index to pokeweed mitogen (PWM), nitric oxide (NO) and lysozyme enzyme activities were measured. Histopathology of spleen was performed. TUM at 2 g kg^?1 significantly (p < .05) improved weight gain, leucocytes, NO and lysozyme activity than control and other treatment groups. IL‐2 level was significantly lower (p < .05) in T2 and T3 than in control and T1, while IL‐4 and lymphocyte proliferation index were significantly (p < .05) elevated in T3 than control, T1 and T2. Spleens of T2 showed improvement in white pulp, while spleen of T3 and T4 revealed white pulp depletion. In conclusion, TUM supplementation exerted immunomodulatory effect in Oreochromis niloticus through manipulation of lymphocyte count, IL‐2, IL‐4 and antibacterial enzymatic activity (NO and lysozyme) that resulted in higher weight gain. Increasing TUM supplementation of fish diets beyond 2 g kg^?1 was not beneficial.     

Dietary taurine improves reproductive performance of Nile tilapia (Oreochromis niloticus) broodstock

This study was conducted to evaluate the effects of supplemental taurine on reproductive performance of Nile tilapia (Oreochromis niloticus) broodstock fed soybean meal‐based diets. Four isonitrogenous (350 g kg^?1 protein), isocaloric (18 MJ kg^?1) diets were formulated to contain 0, 5, 10 and 15 g kg^?1 taurine. The diets were fed to triplicate groups of juvenile Nile tilapia (10–15 g average body weight) at a female: male ratio of 3 : 1, to apparent satiation, three times per day for 130 days. The size at first maturation decreased with increasing dietary taurine to 10 g kg^?1 and levelled off with further taurine supplementation. The time to first spawning was also significantly shorter at 10 g kg^?1 taurine level. Spawning performances, including spawning frequencies, total number of spawnings per tank, number of spawnings per female and absolute fecundity, were all significantly improved with increasing dietary taurine up to 10 g kg^?1. However, the quadratic regression analyses indicated that the maximum spawning performance occurred at 8 g kg^?1 of supplemental taurine. Eggs produced from broodstock fed 10 g kg^?1 taurine exhibited significantly higher hatchability and required shorter time for hatching and yolk‐sac absorption and also resulted in higher larval weight than at other dietary taurine levels. The highest egg protein, total amino acids and taurine were also obtained at 10 g kg^?1 taurine. These results suggest that 8 g kg^?1 dietary taurine is required for optimum reproductive outputs of Nile tilapia broodstock.     

Ultraviolet-induced androgenesis in Nile tilapia, Oreochromis niloticus (L.), and hybrid Nile × blue tilapia, O. aureus (Steindachner)

Fertilization of ultraviolet (UV) irradiated oocytes of Nile tilapia, Oreochromis niloticus (L.), with sperm from O, niloticus or blue tilapia, O, aureus (Steindachner), and subsequent suppression of the first cleavage of fertilized eggs successfully induced androgenesis in Nile and blue tilapia. The optimal doses of UV irradiation to denucleate a female genome of Nile tilapia prior to androgenesis ranged from 5940 to 6930 erg mm^?2 for 54-63 s at a fixed intensity of 110 erg mm^?2 s^?1. Putative androgenetic fish were created from eggs which were irradiated at various times and several durations of heat-shock. Eggs which were treated for 5 min at 41.6 ^oC at 2 7.5 min after fertilization were the most successful at suppressing the first cleavage and producing viable androgenetic diploids in Nile or hybrid Nile X blue tilapia. The maximal survivals of putative androgenetic diploids in relation to the control were 1.60% and 0.90% in Nile and hybrid Nile X blue tilapia, respectively. The androgenetic offspring established exhibited active feeding and normal growth.     

尼罗罗非鱼卵黄脂磷蛋白的分离纯化与性质鉴定

采用Sephacryl S-300过滤层析和DEAE-Sepharose Fast Flow离子交换层析相结合的方法从尼罗罗非鱼(Oreochromis niloticus)成熟卵子匀浆液中分离纯化出了一种高分子量的蛋白。该蛋白能被Schiff试剂、甲基绿和苏丹黑B着色,Western blot显示能被金鱼卵黄脂磷蛋白(lipovitellin,Lv)多克隆抗血清特异性识别,在非变性条件下分子量约为560 k D,在SDS变性条件下分子量约为112 k D,结果表明分离纯化的蛋白是一种含有糖、磷、脂基团的蛋白,符合鱼类Lv的性质,且与金鱼Lv有免疫交叉反应,从蛋白的性质和免疫原性以及分子量大小等角度判断,本研究获得的高纯度蛋白为尼罗罗非鱼卵黄脂磷蛋白;纯化的罗非鱼Lv在反复冻融、37℃及60℃处理条件下均未出现降解,表明罗非鱼Lv比鱼类卵黄原蛋白(Vitellogenin,Vtg)更为稳定。研究结果为罗非鱼Lv抗体的制备奠定了基础。