RAPD-SCAR Markers for Genetically Improved NEW GIFT Nile Tilapia (Oreochromis niloticus niloticus L.) and Their Application in Strain Identification

The NEW GIFT Nile tilapia （Oreochromis niloticus niloticus L.） is a nationally certificated new strain selected over 14 years and 9 generations from the base strain of GIFT Nile tilapia, introduced in 1994. This new variety has been extended in most of areas of China. The management of genetically improved strains, including the genetic markers for identification is needed urgently. RAPD analysis was conducted and their conversion to SCAR markers was developed. From NEW GIFT Nile tilapia, two strain-specific RAPD bands, S304624 bp and S36568 bp were identified. The strain-specific RAPD bands were gel-purified, cloned, and sequenced. Locus-specific primers were then designed to amplify the strain-specific bands. PCR amplification was conducted to test the variations in allele frequencies of two converted SCAR markers among the NEW GIFT Nile tilapia and its base strains, as well as 7 additional farmed strains worldwide. The frequency of SCAR marker Ⅰ （553 bp） was 85.7% in NEW GIFT Nile tilapia, but 16.7% in the base strain. The frequency of SCAR marker Ⅱ （558 bp） was 91.4% in NEW GIFT Nile tilapia, but 0% 70% in the 7 other strains. In order to confirm the utility of these two markers, an examination was conducted for a wild population from Egypt, resulted the frequency of SCAR Ⅰ and Ⅱ was 10% and 70%, respectively, much lower than that of New GIFT strain. The increase in allele frequency of these two SCAR markers suggests that these markers might be genetically linked to the quantitative trait loci （QTL） underlining the performance traits by long term selection, and indicate the bright potential of SCAR marker technology for tracking generations during selection progress and for distinguishing among genetically improved strain and other strains.

仿生态条件下星洲红鱼(Singapore red tilapia)和奥尼罗非鱼(Oreochromis niloticus)生长和肌肉营养特性的比较研究

为比较星洲红鱼(Singapore red tilapia)和奥尼罗非鱼(Oreochromis niloticus)的生长和肌肉营养特性,实验利用两种鱼的不同体色将其混养在同一池塘中,在相同的环境条件和养殖管理下研究其生长特性,并分析肌肉营养成分。结果表明:两种罗非鱼生长在前60 d无差异,60 d后星洲红鱼的体长、体质量日增长和特定生长率均大于奥尼罗非鱼,且差异显著(P<0.05)。据此得到了星洲红鱼和奥尼罗非鱼的体长(L)、体质量(W)生长与养殖天数(d)的回归方程。星洲红鱼含肉率高于奥尼罗非鱼9.65%(P<0.05),肌肉水分高于奥尼罗非鱼,蛋白质、粗脂肪和灰分含量均低于奥尼罗非鱼,其中脂肪含量差异显著(P<0.05),其他不显著。两种罗非鱼肌肉的氨基酸组成全面,氨基酸组成均符合FAO/WHO的理想模式,除精氨酸含量有差异外,其他氨基酸含量稍有差异但不显著。星洲红鱼和奥尼罗非鱼肌肉EAA/TAA、EAA/NEAA、DAA/TAA和EAAI分别为41.04%和40.95%,70.00%和69.34%,40.71%和39.08%,67.46%和67.08%,差异不显著。星洲红鱼、奥尼罗非鱼肌肉∑SFA和∑UFA分别为28.03%和28.60%,63.87%和61.38%,各脂肪酸含量除C14:0和C18:2外,其他含量差异不显著。研究表明,星洲红鱼和奥尼罗非鱼生长快、营养丰富、食用价值高,是优质淡水鱼类养殖品种,其中星洲红鱼含肉率高,生长快,养殖效益明显。

Comparative Study on the Antioxidant Activity of Peptides from Pearl Oyster(Pinctada martensii) Mantle Type V Collagen and Tilapia(Oreochromis niloticus) Scale Type Ⅰ Collagen

In this study, Pearl oyster mantle type V collagen(POMC) and tilapia scale type I collagen(TSC) were extracted and hydrolyzed by various proteases in order to obtain peptides. The antioxidant activity of the peptides was investigated by DPPH, hydroxyl radical scavenging experiments and a dynamic digestion model in vitro. The results show that there are significant differences in amino acid composition between POMC and TSC. The collagen peptides obtained from pearl oyster mantle(POMCP) by treating with alkaline protease exhibited higher antioxidant activity than that from tilapia scale(TSCP) treated with papaya protease, and both of them showed greater DPPH and hydroxyl radical scavenging activity than other peptides. After being separated via Sephadex G-25 chromatography, the M1 fraction isolated from POMCP, and the S1 fraction from TSCP with which both had higher molecular weights showed the strongest antioxidant activity than other fractions, and the M1 fraction exhibited stronger antioxidant activity than the S1 fraction in scavenging free-radicals and protecting cells from the oxidation damage. Furthermore, after treating the dynamic digestion system model in vitro, the DPPH and hydroxyl radical scavenging activity of the M1 fraction increased slightly. These results suggest that POMCP exhibits stronger antioxidant activity than TSCP, which means that PMOP may be a good candidate to be a potential natural antioxidant in the food-processing industry.

Effects of phenanthrene on hepatic enzymatic activities in tilapia(Oreochromis niloticus ♀×O.aureus ♂)

The effects of phenanthrene（Phe） on hepatosomatic index（HSI） and hepatic enzymatic activities in hybrid tilapia（Oreochromis niloticus ♀× O.aureus ♂） were investigated via the static freshwater exposure at dosage of 50,100,and 400 μg/L for 4-14 d.Compared with the control group,HSI was significantly decreased（P 〈 0.05） at 400 μg/L at day 14.Increased enzymatic activities（P 〈 0.05） were observed for catalase（CAT）,glutathione peroxidase（GPx） and superoxide dismutase（SOD） at either 100 or 400 μg/L at day 8 and 14,as well as for CAT at 50 μg/L at day 14,except for GPx at 400 μg/L at day 8.Ethoxyresorufin O-deethylase（EROD） activity was significantly increased（P 〈 0.05） at all dosage at day 4 as well as at 50 μg/L at day 8,but significantly decreased at either 100 or 400 μg/L at day 14（P 〈 0.05）.Glutathione-S-transferase（GST） activity was not affected.The results suggest that CAT,GPx,SOD and EROD,as well as HSI in tilapia may be used as the biomarkers or indexes for evaluating or monitoring the pollution of polycyclic aromatic hydrocarbons（PAHs） such as Phe.

Behavioral response of tilapia (Oreochromis niloticus) to acute ammonia stress monitored by computer vision

The behavioral responses of a tilapia (Oreochromis niloticus) school to low (0.13 mg/L), moderate (0.79 mg/L) and high (2.65 mg/L) levels of unionized ammonia (UIA) concentration were monitored using a computer vision system. The swimming activity and geometrical parameters such as location of the gravity center and distribution of the fish school were calculated continuously. These behavioral parameters of tilapia school responded sensitively to moderate and high UIA concen-tration. Under high UIA concentration the fish activity showed a significant increase (P<0.05), exhibiting an avoidance reaction to high ammonia condition, and then decreased gradually. Under moderate and high UIA concentration the school’s vertical location had significantly large fluctuation (P<0.05) with the school moving up to the water surface then down to the bottom of the aquarium alternately and tending to crowd together. After several hours’ exposure to high UIA level, the school finally stayed at the aquarium bottom. These observations indicate that alterations in fish behavior under acute stress can provide important in-formation useful in predicting the stress.

Induction of testis-ova in nile tilapia (Oreochromis niloticus) exposed to 17β-estradiol

The efficacy of Endocrine Disrupting Compounds (EDCs), 17β-estradiol was tested on the fish Oreochromis niloticus in order to understand the intersex relationship of fish, in which sequential hermaphrodism can consist of a male changing into a female (protandry) or a female changing into a male (protogyny). The fish were equally divided into 3 groups. The first group was the control group;the second and third groups were treated with 10 and 100 mg L-1 of 17β-estradiol, respectively, for 30 days. The overall result in this experiment had no significant effect on the growth parameters. Among the two treated groups, the low concentration group shows results similar to those of the control groups. The high concentration group shows changes to the male reproductive system with the appearance of the testis-ova present resulting in an intersex condition of the male gonads. With this experiment, it can be concluded that 17β-estradiol at high concentration reveals positive changes towards the male reproductive system of the fish, Oreochromis niloticus.

Dietary L-arginine supplementation reduces lipid accretion by regulating fatty acid metabolism in Nile tilapia(Oreochromis niloticus)

Background: Excessive white fat accumulation in humans and other animals is associated with the development of multiple metabolic diseases. It is unknown whether dietary L-arginine supplementation reduces lipid deposition in high fat diet-fed Nile tilapia(Oreochromis niloticus).Results: In the present study, we found that dietary supplementation with 1% or 2% arginine decreased the deposition and concentration of fats in the liver;the concentrations of triglycerides, low-density lipoprotein, total cholesterol, and high-density lipoprotein in the serum;and the diameter of adipocytes in intraperitoneal adipose tissue. Compared with the un-supplementation control group, the hepatic activities of alanine aminotransferase,aspartate aminotransferase, and lactate dehydrogenase, and hepatic concentration of malondialdehyde were reduced but these for catalase and superoxide dismutase were enhanced by dietary supplementation with 2% arginine. Arginine supplementation reduced the total amounts of monounsaturated fatty acids, while increasing the total amounts of n-3 and n-6 polyunsaturated fatty acids in the liver. These effects of arginine were associated with reductions in mRNA levels for genes related to lipogenesis(sterol regulatory element-binding protein-1, acetyl-CoA carboxylase α, stearoyl-CoA desaturase, and fatty acid synthase) but increases in mRNA levels for genes involved in fatty acid β-oxidation(carnitine palmitoyltransferase 1α and peroxisome proliferator-activated receptor α). In addition, hepatic mRNA levels for Δ4 fatty acyl desaturase 2 and elongase 5 of very long-chain fatty acids were enhanced by arginine supplementation.Conclusion: These results revealed that dietary L-arginine supplementation to tilapia reduced high fat diet-induced fat deposition and fatty acid composition in the liver by regulating the expression of genes for lipid metabolism.

Histopathological alterations in the liver and intestine of Nile tilapia Oreochromis niloticus exposed to long-term sublethal concentrations of cadmium chloride

Fingerlings of Nile tilapia Oreochromis niloticus were exposed to 1.68, 3.36, and 5.04 mg/L cadmium(as CdCl2), which represent 10%, 20%, and 30% of their previously determined 96-h LC5 0. After exposure for 20 days, sections of the liver and intestine of treated fish were examined histologically. Histopathological changes varied from slight to severe structural modification, depending on the exposure concentration. The hepatic tissues of fish exposed to 10% LC 50 showed markedly increased vacuolation of the hepatocytes and coarse granulation of their cytoplasm. Abundant erythrocytic infiltration among the hepatocytes was observed in fish exposed to 20% LC5 0. In the intestinal tissues of fish exposed to all doses, goblet cells proliferated and were greatly increased in size, the longitudinal muscularis mucosa was disturbed and, in the crypts of the sub-mucosal layer, apoptosis increased, indicated by large numbers of degenerated nuclei. Large numbers of inflammatory cells and dilated blood vessels were observed in the intestine of the group treated with 30% LC5 0.

Effect of dietary genistein on growth performance,digestive enzyme activity,and body composition of Nile tilapia Oreochromis niloticus

An 8-week feeding experiment was performed to evaluate the effect of dietary genistein on growth performance,body composition,and digestive enzymes activity of juvenile Nile tilapia(Oreochromis niloticus).Four isonitrogenous and isoenergetic diets were formulated containing four graded supplements of genistein:0,30,300,and 3 000 μg/g.Each diet was randomly assigned in triplicate to tanks stocked with 15 juvenile tilapia(10.47±1.24 g).The results show that 30 and 300 μg/gdietary genistein had no significant effect on growth performance of Nile tilapia,but the higher level of genistein(3 000 μg/g)significantly depressed the final body weight and specific growth rate.There was no significant difference in survival rate,feed intake,feed efficiency ratio or whole body composition among all dietary treatments.An assay of digestive enzymes showed that the diet containing 3 000 μg/ggenistein decreased stomach and hepatopancreas protease activity,and amylase activity in the liver and intestine,while a dietary level of300 μg/g genistein depressed stomach protease and intestine amylase activities.However,no significant difference in stomach amylase activity was found among dietary treatments.Overall,the results of the present study indicate that a high level of dietary genistein(3 000 μg/g,or above) would significantly reduce the growth of Nile tilapia,partly because of its inhibitory effect on the activity of major digestive enzymes.Accordingly,the detrimental effects of genistein,as found in soybean products,should not be ignored when applied as an alternative ingredient source in aquaculture.

Effect of Ammonia-Nitrogen Stress on Non-specific Immunity of Hybrid Tilapia(Oreochromis niloticus× Oreochromis areus)

The juveniles of hybrid tilapia （ Oreochromis niloticus x Oreochromis areus） were exposed to ananonia-nitrogen （N） （0, 2.5, 5, 10 and 20 mg/L） for 0, 12, 24, 48 and 72 h to evaluate the effect of ammonia-N stress on their non-specific immunity. Results show that the activity of serum lysozyme decreased signifi- candy with extension of stress time （P 〈 0.05）. The total antioxidant capacity （T-AOC） and activity of antioxidase in liver were significantly affected. The activi- ty of T-AOC and total superoxide dismutase （T-SOD） of fish exposed to ammonia-N were initially decreasing then increasing （ P 〈 0.05 ）. Activities of catalase （CAT） and glutathione peroxidase （GSH-Px） were correlated with concentrations of ammonia-N. Fish exposed to lower concentrations （2.5 mg/L or 5 mg/L） showed decreased CAT activity within 24 h （P 〈 0.05 ）, while those exposed to higher concentrations （10 mg/L or 20 mg/L） initially showed increased then decreased activity of CAT. Except for the highest concentration groups, fish exposed to ammonia-N showed inductive activity of GSH-Px （ P 〈 0.05 ）. Under the ex- perimental conditions, non-specific immunity of tilapia was affected by ammonia-N stress, and the impact was increased with increased concentration and extension of time.

Molecular cloning and expression analysis of FTZ-F1 in the GIFT tilapia,Oreochromis niloticus

The FTZ-F1 genes encode orphan receptors of the nuclear receptor superfamily and in mammals have been found to play important roles in the proper development of the adrenal-gonadal axis and sex-determination.We isolated the homologue of FTZ-F1 in genetically improved farmed tilapia(gfFTZ-F1).The full-length cDNA was isolated from the ovary,which included an open reading frame encoding a predicted protein of 486 amino acids.Sequence,tissue distribution and phylogenic analysis of the FTZ-F1 showed that the gfFTZ-F1 belonged to SF-1/Ad4BP group and that gfFTZ-F1 transcripts were only expressed in the gonads and kidney but not in other tissues.Likewise our data suggests that the gfFTZ-F1 gene may share similar functions with other fish and mammalian counterparts,though further study is needed to make any definitive conclusions.

Phylogenetics and Molecular Divergence of Tilapia Fish (Oreochromis Species) Using Mitochondrial D-Loop and Cytochrome b Regions

Understanding the level of genetic diversity in any population is an important requisite towards strategizing measures for conservation and improvement of stocks. This study focused on the assessment of phylogenetics and molecular divergence of tilapia fish species obtained from two populations (Domita in South-South and Odeda in South-West, Nigeria) using the displacement loop (D-loop) and cytochrome b region of the mitochondrial deoxyribonucleic acid (mtDNA). A total of 28 samples (15 from South-South and 13 from South-West) were used for the genetic analysis. DNA was extracted from the tissue of all the samples using Quik-gDNATM miniPrep kit. The D-loop containing the hypervariable region was sequenced for all samples from the two populations, while cytochrome b (Cyt b) region of mtDNA was only sequenced for samples from South-South population. Chromatograms of the sequences were viewed and edited using Bioedit software. Multiple sequence alignment was carried out using molecular evolutionary genetic analysis (MEGA) software before subsequent genetic analyses. Phylogenetic analysis grouped the samples into two clusters based on population. Also, when the two mitochondrial regions were pooled together, they clustered into two major groups based on mitochondrial regions. Analysis of molecular variance (AMOVA) revealed 37.32% variation within population and 62.68% variation among population with a significant fixation index of 0.627 (p 0.05). The genetic distance inferred between D-loop regions of South-South and South-West populations was 0.243. Maternal lineage analysis revealed that the origin of tilapia fish from both populations could be traced to Oreochromis spirilus and Oreochromis leucostictus based on mitochondrial D-loop region. The findings of this study revealed molecular divergence among the tilapia populations and may serve as pivot information for the genetic improvement of this important species.

REDUCED GROWTH IN HYBRID TILAPIA (OREOCHROMIS MOSSAMBICUS×O. NILOTICUS) AT INTERMEDIATE STOCKING DENSITY

Hybrid tilapia were reared at densities of 1, 5 or 10 fish per tank for four weeks. Mortality was 0 at 1 and 10 fish per tank, but was 25% at 5 fish per tank. Specific growth rate was highest at 1 fish per tank, and lowest at 5 fish per tank. The lower growth rate at the intermediate stocking density was associated with reduced feed efficiency, but there was no reduction in feed intake or digestibility. The results suggested increased metabolic cost caused by aggressive behaviour at intermediate stocking density, which can be suppressed by a further increase in density.

Compensatory growth in hybrid tilapia (Oreochromis mossambicus×O. niloticus) reared in seawater, following restricted feeding

Hybrid tilapia weighing 7.71 g were reared in seawater at 24.0-29.0℃ for 8 weeks. The controls were fed to satiation twice a day throughout the experiment, whereas treatment groups were fed at 0.5%, 1.5% or 3.0% body weight per day for 4 weeks, and then to satiation for the remainder of the experiment. During the first 4-week period, there was a curvilinear relationship between growth rate and ration size. Fish fed 0.5% and 1.5% rations displayed compensatory growth response of 2 weeks duration during realimentation. The weight-adjusted growth rate of fish fed at 3% ration was not significantly different from that of the controls by the end of the experiment, when none of the treatment groups had caught up in body weight with the controls. Hyperphagia was observed for the first 2 weeks of realimenatation in fish previously fed at 3% ration, but persisted for the whole realimentation period in groups previously fed at 0.5% and 1.5% rations. None of the feed restricted groups showed improved digestibility, feed efficiency, or protein and energy retention efficiency.

Effects of dietary genistein on GH/IGF-I axis of Nile tilapia Oreochromis niloticus

There is considerable concern that isoflavones,such as genistein in fish feed composed of soybean protein,affects somatic growth in fish.Our previous works demonstrated that 30 and 300 μg/g dietary genistein had no significant effect on growth performance in Nile tilapia(Oreochromis niloticus),but the higher level of genistein(3 000 μg/g) significantly depressed growth.This study was conducted to further examine the effects of dietary genistein on the endocrine disruption on growth hormone/insulin-like growth factor-I(GH/IGF-I) axis in Nile tilapia(O.niloticus).Juvenile fish were fed by hand twice daily to satiation with one of four isonitrogenous and isoenergetic diets,each containing either 0,30,300 or3 000 μg/g genistein.Following an 8-week feeding period,plasma GH and IGF-I levels were investigated by radioimmunoassay and gene expression levels of gh,ghrelin,gnrhs,ghr,npy,npyrs,pacap,ghrs,igf-I,igf-Ir,and igfbp3 were examined by real-time PCR.The results show that no significant change in plasma GH and IGF-I levels in fish fed with diets containing 30 μg/gand 300 μg/g genistein.mRNA expression of genes along the GH/IGF-I axis remained unaffected,except for igf-Ir,which was stimulated by the 300 μg/g genistein diet.While in fish fed the 3 000 μg/g genistein diet,the plasma GH and IGF-I levels decreased,and mRNA expression of gh,ghr2,npyr1,igf-I,and igf-Ir were also significantly depressed.In contrast,npy and igfbp3 mRNA expression were enhanced.This study provides convincing evidence for growth impediment by genistein by disturbing the GH/IGF-I axis in Nile tilapia O.niloticus.

Chitosan Extends the Shelf-life of Filleted Tilapia (Oreochromis niloticus) During Refrigerated Storage

Shelf-life extension of aquatic products is of significant economical importance. To determine the potential effect of chitosan on the shelf-life of filleted tilapia, this study analyzed the bacterial community diversity in fresh and spoiled tilapia fillets stored at (4 ± 1)℃ and examined the antimicrobial activity of chitosan against relevant bacteria isolates. Results showed that Pseudomonas (20%) and Aeromonas (16%) were abundant in fresh tilapia fillets, whereas Pseudomonas (52%), Aeromonas (32%) and Staphylococcus (12%) were dominant in the spoiled samples. Chitosan showed wide-spectrum antibacterial activity against bacteria isolated from tilapia and 5.0 g L-1 chitosan was selected for application in preservation. We further determined the shelf-life of chitosan-treated, filleted tilapia stored at (4 ± 1)℃ based on microbiological, biochemical and sensory analyses. Results showed that the shelf-life of chitosan-treated, filleted tilapia was extended to 12 d, whereas that of untreated, control samples was 6 d. These indicate that chitosan, as a natural preservative, has great application potential in the shelf-life extension of tilapia fillets.

Effect of Using Baker＇s Yeast and Exogenous Digestive Enzymes as Growth Promoters on Growth, Feed Utilization and Hematological Indices of Nile tilapia, Oreochromis niloticus Fingerlings

The present study was conducted to evaluate the effect of baker yeast, Saccharomyces cerevisiae （SC） and exogenous digestive enzymes （pepsin, papain and a-amylase, EDE） dietary supplementation on growth performance, feed utilization and hematological indices of Nile tilapia, Oreochromis niloticus fingerlings. A total of 630 Nile tilapia fingerlings with an average body weight of 26.4 ± 0.2 g were divided in the seven experimental net-pen treatments （three replicates each）. The experiment was conducted for 119 days. Seven isonitrogenous （26.50%） digestible protein and isocaloric （13.40 MJ kgl） digestible energy experimental diets were formulated. The control diet had no SC and EDE added. Diets 2-3 each contained SC at levels of 2 and 4 g 100 g diet-t, respectively, while diets 4-5 each contained EDE at levels of （0.64, 1.28, 0.16） and （1.28, 2.56, 0.32） g 100 gdiefI of pepsin, papain and a-amylase, respectively. Diet 6 contained mixture of SC and EDE at levels of 1 g yeast and 0.32, 0.64, 0.08 g of pepsin, papain and a-amylase, respectively 100 gdiet1 and diet D7 contained 2 g yeast and 0.64, 1.28, 0.16 g of pepsin, papain and a-amylase, respectively 100 g dietl. Growth performance and feed utilization efficiency of Nile tilapia were significantly （P 〈 0.05） higher in all treatments receiving SC and/or EDE supplemented-diets than the control diet which suggests that the addition of SC and EDE enhanced the growth performance. Red blood cells counts, hematocrit and hemoglobin were significantly （P 〈 0.05） highest in all treatments receiving mixture of SC and EDE supplemented-diets （D6 ＋ D7）. The same trend was observed for total plasma protein and total plasma globulin levels. The results of present study suggested that Nile tilapia fingerlings fed diets containing the mixture of I g yeast, SC and 0.32, 0.64, 0.08 g of pepsin, papain and a-amylase, respectively 100 gdiet^-1, for 119 days had enhanced growth performance, diet utilization efficiency and hematological indices.

Growth performance,digestive enzyme activities and serum nonspecific immunity of the red tilapia(Oreochromis mossambicus×Oreochromis niloticus)feddiets supplemented with ultrafine powder of Enteromopha prolifera

The present study evaluated ef fects of ultrafine powder of the green macroalgae E nteromopha prolifera as dietary supplement on growth performance, digestive enzyme activities and serum nonspecific immune responses of the red tilapia(O reochromis mossambicus × Oreochromis niloticus). The red tilapia were fed five diets supplemented with different levels of E. prolifera ultrafine powder as well as a control diet containing no E. prolifera for seven weeks(Diets 0–6 contained 0(control), 10, 20, 30, 40 and 50 g/kg of E. prolifera ultrafine powder, respectively). The results showed that diets supplemented with E. prolifera ultrafine powder generally improved growth, immunity and digestive enzyme activities of the red tilapia. In particular, the fish fed the diet incorporated 50 g/kg (5%) E. prolifera ultrafine powder(Diet 5) achieved the highest percentage weight gain, specific growth rate and the condition factor(increased by 15.4%, 8.0% and 5.7%, respectively when compared to the control). Feeding the diet also led to significantly increases( P <0.05) in serum nonspecific immune responses, including total superoxide dismutase, acid phosphatase, alkaline phosphatase, lysozyme activities and serum total protein(increased by 19.4%, 48.1%, 29.5%, 30.3% and 8.7%, respectively) as well as digestive enzyme activities of erepsin, gastric amylase, gastric lipase, pepsin, intestinal amylase and gastric lipase(increased by 15.7%, 33.3%, 16.3%, 21.3%, 52.3% and 28.2%, respectively) than those of the control. Based on these results, it is recommended that the inclusion level of E. prolifera ultrafine powder in the diet of the red tilapia should be 50 g/kg(or 5%).

Variation in metabolic enzymatic activity in white muscle and liver of blue tilapia,Oreochromis aureus,in response to long-term thermal acclimatization

The effects of rearing temperature on white muscle and hepatic phosphofructokinase （PFK）, pyruvate kinase （PK）, lactate dehydrogenase （LDH）, aspartate aminotransferase （AST） and alanine aminotransferase （ALT） were examined in fingerlings of blue tilapia, Oreochromis aureus. The experiment was conducted for 14 weeks at temperatures of 18, 22, 26, 30, and 34℃. The activity of the glycolytic enzymes PFK, PK, and LDH in white muscle increased significantly with increase in water temperature. A reverse trend was observed for these enzymes in the liver, except for LDH, which behaved in the same manner as in white muscle. Cytosolic AST and ALT activity increased in both white muscle and liver in response to warm thermal acclimatization, while a reduction in mitochondrial AST and ALT activity was noticed at high temperatures in comparison with those at a lower temperature.

Evaluation of Nitrogen and Phosphorus Wastes Produced by Nile Tilapia (<i>Oreochromis niloticus</i>L.) Fed <i>Azolla</i>-Diets in Earthen Ponds

Nitrogen (N) and phosphorus (P) wastes produced by Nile tilapia Oreochromis niloticus L. fed Azolla, an aquatic atmospheric nitrogen fixing fern, was evaluated for 90 days in pond experiment. Six isonitrogenous (29.2% crude protein) and isoenergetic (16.9 Kj·g–1) diets A0, A10, A20, A30, A40 and A50, containing 0%, 10%, 20%, 30%, 40% and 50% of Azolla meal (AM) respectively, as partial fishmeal (FM) substitutes, was provided to experimental fish. The Azolla-free diet A0 served as a control. Fish specific growth rate (SGR) was higher with the control diet, the lower values being obtained in A50-fed fish (P Azolla could be used in diet to sustain Nile tilapia growth and as “environmentally-friendly” ingredient to limit P loss, while providing N to the field, beneficially in tropical marshland pond where this nutrient is already limiting.