Production of a high percentage of male offspring in growth-enhanced transgenic tilapia using Oreochromis aureus ZZ selected pseudofemales

Gene transfer has offered a new tool for the development of improved fish strains for aquaculture. Monosex fish populations could minimize the impact of genetically modified organisms in the environment. In Oreochromis aureus, the use of pseudofemale spawners (sex-reversed male with a female phenotype) is an alternative technique for producing genetically male tilapia offspring. O. aureus fry were treated with 17β-estradiol at 100 mg/kg of food for 45 days. We obtained 77.1% females and 45.9% in the control group. Females randomly taken from the treated group were crossed with normal males. Fry from pseudofemales producing more than 90% male progenies were submitted to 17 β-estradiol treatment to obtain F₂ pseudofemales. The results of the sex-reversal were low and variable ranging between 66.0 and 84.3% females. F₂ pseudofemales were crossed with transgenic males from the F70 line (O. aureus × O. urolepis hornorum) and non-transgenic (O. aureus) males. The sex ratio of progeny of F₂ pseudofemale deviated significantly (P < 0.01) in favor of males in the crosses with transgenic (90.2%) and non-transgenic (89.3%) males compared to the results observed with normal females (51.0 and 52.3%, respectively). The mean fry production with pseudofemales (per m²/day) was similar to the normal females in the crosses with transgenic and non-transgenic males. To our knowledge this is the first report on the production of a near monosex population in genetically modified fish.     

Effects of dietary starch and energy levels on maximum feed intake, growth and metabolism of Nile tilapia, Oreochromis niloticus

The aim of this study was to gain insight into how Nile tilapia (Oreochromis niloticus) regulate feed and energy intake in response to diets low and high in starch and cellulose. It was hypothesized that high-starch diets would reduce feed intake due to the effect of high blood glucose level, and that stomach volume may limit feed intake of fish fed diets low in energy. Four experimental diets, low starch–no cellulose inclusion, high starch–no cellulose inclusion, low starch–with cellulose inclusion, and high starch–with cellulose inclusion, were formulated. The high-starch diets and diets with cellulose inclusion were 17.5% more energy-diluted than the low-starch diets and diets without cellulose inclusion, respectively. Male tilapia were fed to apparent satiation for six weeks. Feed and digestible energy intake of fish fed diets with cellulose inclusion increased and decreased by 8.3% and 5.5%, respectively, compared to fish fed diets without cellulose inclusion. This suggests the role of stomach volume in restricting feed consumption. Fish fed high-starch diets achieved only 0.5% more feed intake and 13.9% less digestible energy intake than fish fed low-starch diets. The lower increase in feed intake and higher decrease in digestible energy intake of fish fed high-starch diets than of fish fed diets with cellulose inclusion suggests that high blood glucose suppresses feed intake in Nile tilapia. An alternative explanation for the differences in feed and digestible energy intake of fish fed different diets was based on the fact that heat production was not influenced by starch nor cellulose-inclusion levels. Thus, under satiation feeding, oxygen uptake capacity may determine feed and digestible energy intake in fish rather than blood glucose or stomach volume.     

维生素E和硒对吉富罗非鱼(Oreochromis niloticus)幼鱼生长及血清抗氧化酶活性的影响

本研究采用2×5双因子试验设计,即在基础饲料中分别添加0和0.15 mg/kg的Se,每一Se水平下分别添加0、30、60、90、120 mg/kg的维生素E(VE),共制成10种试验饲料,饲喂平均初始体质量为(0.37±0.01)g的吉富罗非鱼(Oreochromis niloticus)70 d,每组设3个重复,每个重复组50尾吉富罗非鱼,其中饲喂未添加Se和VE的基础饲料组为对照组。结果表明,(1)VE对吉富罗非鱼幼鱼增重率、特定生长率、摄食量和饲料系数均有显著影响(P0.05),就生长性能而言,单独添加VE的适宜范围是63.86–70.58 mg/kg。Se对吉富罗非鱼的增重率、特定生长率的影响不显著(P0.05),但在VE添加量为0 mg/kg的试验组中,硒的添加使吉富罗非鱼增重率高于对照组。VE和Se对吉富罗非鱼生长的交互作用没有显著影响(P0.05)。(2)VE对谷胱甘肽过氧化物酶(GSH-PX)和过氧化氢酶(CAT)活性有显著影响(P0.05),总超氧化物歧化酶(T-SOD)和CAT活性随VE添加量的增加呈上升趋势。而Se可显著影响T-SOD和GSH-PX的活性(P0.05),在VE添加量范围60–90 mg/kg时,加Se试验组的T-SOD、GSH-PX和CAT活性较不加Se组有所上升。VE和Se对GSH-PX活性的影响具有显著的交互作用(P0.05)。在本研究条件下,VE和Se对吉富罗非鱼的生长及抗氧化有一定促进作用,二者联合使用,没有协同促生长作用,但抗氧化作用得到进一步加强。     

Effects of dietary carbohydrate level on growth and body composition of juvenile tilapia, Oreochromis niloticus×O. aureus

A growth trial was conducted to feed juvenile tilapia (initial weight, 9.1±0.1 g), Oreochromis niloticus×O. aureus, isonitrogenous diets for 8 weeks. Six diets were formulated containing 29% crude protein from casein and gelatin, 10% crude fat from soybean oil and refined soybean lecithin and varying levels of corn starch ranging from 6% to 46% at increments of 8%, with corresponding energy to protein (E/P) ratios of 35.6, 37.9, 40.2, 42.5, 44.8 and 47.1 kJ g^?1. Weight gain (WG), specific growth rate, feed efficiency ratio and protein efficiency ratio were significantly higher in fish fed diets with starch ≥22% (or E/P ratio ≥40.2 kJ g^?1) than in fish fed diets with 6% or 14% starch (or E/P ratio of 35.6 or 37.9 kJ g^?1). No further improvement was measured when dietary starch content increased beyond 22%. Body protein retention showed the same general pattern as WG, and was highest in fish fed the 22% starch diet. Body composition was significantly affected by dietary starch level. Fish fed diets with starch ≥30% had significantly higher lipid content than fish fed diets with 6% or 14% starch. Ash content was negatively correlated with starch inclusion level, but moisture and protein contents did not show discernible trends among treatments. Results indicate that hybrid tilapia can utilize 46% dietary starch without growth retardation, while 22% starch in feed for juvenile tilapia containing 29% protein and 10% lipid, or an E/P ratio of 37.9 kJ g^?1 is optimal.     

Behavioral responses of tilapia (Oreochromis niloticus) to acute fluctuations in dissolved oxygen levels as monitored by computer vision

Behavioral and ventilatory parameters have the possibility of predicting the stress state of fish in vivo and in situ. This paper presents a new image-processing algorithm for quantifying the average swimming speed of a fish school in an aquarium. This method is based on the alteration in projected area caused by the movement of individual fish during frame sequences captured at given time intervals. The image enhancement method increases the contrast between fish and background, and is thus suitable for use in turbid aquaculture water. Behavioral parameters (swimming activity and distribution parameters) and changes in ventilation frequency (VF) of tilapia (Oreochromis niloticus) responded to acute fluctuations in dissolved oxygen (DO) which were monitored continuously in the course of normoxia, falling DO level, maintenance of hypoxia (three levels of 1.5, 0.8 and 0.3 mg l⁻¹) and subsequent recovery to normoxia. These parameters responded sensitively to acute variations in DO level; they displayed significant changes (P < 0.05) during severe hypoxia (0.8 and 0.3 mg l⁻¹ level) compared with normoxic condition, but there was no significant difference under conditions of mild hypoxia (1.5 mg l⁻¹ level). There was no significant difference in VF between two levels of severe hypoxia 0.8 and 0.3 mg l⁻¹ level during the low DO condition. The activity and distribution parameters displayed distinguishable differences between the 0.8 and 0.3 mg l⁻¹ levels. The behavioral parameters are thus capable of distinguishing between different degrees of severe hypoxia, though there were relatively large fluctuations.     

Long-term effects and interactions of dietary vitamins C and E on growth and reproduction of yellow perch, Perca flavescens

This study was conducted to examine the effects of dietary vitamin C and/or E and their interaction on growth and reproductive performance in yellow perch. Yellow perch juveniles were divided into four treatments with triplicate groups and fed with one of four semi-purified diets for 32 weeks. The experimental diets were formulated to contain no or high vitamin E levels (160 mg/kg) without or with vitamin C supplementation (250 mg/kg) designated as diets −C−E, −C+E, +C−E, or +C+E, respectively. The growth rates and survival of yellow perch fed with +C−E and +C+E diets were significantly higher than the fish fed with −C−E diet. Total- and dehydro-ascorbate concentrations in liver and testis increased in response to its dietary supplementation. The -tocopherol concentration in sperm was significantly affected by dietary supplementation of vitamin C. Gonadosomatic index of males was lower in the fish fed with −C−E diet compared to that of fish fed with other diets. Hatching rates of embryos obtained from testing sperm viability was significantly improved in fish fed with the diets sufficient in vitamin C (82±7%) compared to the fish fed with the −C−E diet (59±7). This study indicates that supplementation of dietary vitamin C and E increase growth rate, and semen quality can be improved by dietary vitamin C supplementation within one maturation cycle, and vitamin C may spare sperm vitamin E depending on vitamin E stores in tissues.     

Effect of supplemental dietary chromium and vanadium on the utilization of different carbohydrates in tilapia, Oreochromis niloticus×O. aureus

Diets containing 40% glucose or starch with either 2 ppm chromium (Cr), 2 ppm vanadium (V) or without either supplement were fed to tilapia, Oreochromis niloticus×O. aureus, to investigate the effect of Cr and V on carbohydrate utilization. Each diet was fed to three aquaria with 18 fish/aquarium in a recirculating, filtered rearing system for 8 weeks. Results indicated that weight gain, protein and energy deposition were significantly (P<0.05) higher in fish fed the starch diet than in those fed the glucose diet. Chromium supplementation significantly increased the weight gain, energy deposition and liver glycogen content in fish fed the glucose diet. Delayed plasma glucose plateau and significantly higher body lipid content were observed in fish fed the glucose diet with Cr than in those without the Cr supplementation.     

Effects of dietary cecropin on growth,non‐specific immunity and disease resistance of tilapia (Oreochromis niloticus × O. aureus)

An 8‐week feeding trial was conducted to evaluate the effect of dietary cecropin on growth performance, non‐specific immunity and disease resistance of hybrid tilapia (Oreochromis niloticus × O. aureus). A basal diet was supplemented with 0 (control), 75, 150 and 225 mg kg^?1 cecropin to formulate four experimental diets. The results showed that final body weight and specific growth rate obtained with diet 150 mg kg^?1 cecropin were significantly higher (P < 0.05) than that obtained with diets 0 and 75 mg kg^?1, whereas there was no difference between 150 and 225 mg kg^?1. Feed conversion efficiency was improved by increasing the dietary cecropin content (P < 0.05). However, feeding intake was reduced as dietary cecropin increased. Serum lysozyme, alkaline phosphatase, catalase and superoxide dismutase activity were significantly increased with the increased levels of dietary cecropin (P < 0.05), and reached a maximum at level of 150 mg kg^?1 cecropin, then those values no longer increased. Moreover, the dietary cecropin supplementation level also exhibited a decrease in the cumulative mortality rates compared with the controls when challenged with Aeromonas veronii. The results suggested that the dietary supplementation of cecropin could improve the growth, non‐specific immunity indicators and resistance of hybrid tilapia against A. veronii.     

维生素C对青鱼幼鱼生长、免疫及抗氨氮胁迫能力的影响

以初始体质量为(7.27 ±0.40)g的青鱼为研究对象,采用维生素C(VC)含量分别为0(对照组)、16.3、33.9、69.1、137.8和277.5 mg/kg 6种等氮等能实验饲料,饲养青鱼8周后,选取0、69.1和277.5 mg/kg VC组进行24 h氨氮胁迫(20 mg/L),研究VC对青鱼幼鱼生长、免疫及抗氨氮胁迫能力的影响.结果显示:以特定生长率为指标,折线模型分析表明青鱼有效维生素C需要量为63.0 mg/kg.肌肉、肝脏和血清中VC积累量与饲料中VC含量呈正相关性,当VC添加量达到137.8 mg/kg时,肌肉和肝脏中VC积累量达到饱和.饲料中添加VC对血清总超氧化物歧化酶(T-SOD)活性及丙二醛(MDA)含量无显著影响,饲料中添加VC显著提高了血清谷胱甘肽过氧化物酶(GSH-px)和过氧化氢酶(CAT)活性(P＜0.05);血清中谷胱甘肽(GSH)含量和鳃丝Na+/K+-ATP酶活力(NKA)随饲料中VC添加量的增加呈升高趋势,而血清皮质醇含量(COR)呈下降趋势.氨氮胁迫有使肝脏和肌肉VC含量、血清T-SOD、CAT、GSH-px活性、GSH含量和鳃丝NKA活性降低,血清VC含量、COR和MDA含量增加趋势.其中,对照组T-SOD、CAT、NKA活性、GSH、MDA和COR胁迫前后差异显著(P＜0.05),氨氮胁迫使VC添加组青鱼肝脏VC含量显著降低(P＜0.05),而CAT、NKA活力、GSH、COR胁迫前后均无显著变化.69.1 mg/kg VC组T-SOD活性胁迫前后无显著差异,但GSH-px活性和MDA含量胁迫前后差异显著(P＜0.05),而277.5 mg/kg VC添加组GSH-px活力和MDA含量胁迫前后差异不显著,但T-SOD活性显著降低,血清VC含量显著升高(P＜0.05).研究表明,青鱼获得最好生长的饲料有效VC添加量为63.0 mg/kg,氨氮胁迫使青鱼产生免疫应激反应,而补充VC可有效增强机体免疫,缓解机体免疫应激,改善青鱼抗氨氮胁迫能力.     

Effects of totally replacing dietary fish oil by linseed oil or soybean oil on juvenile hybrid sturgeon,Acipenser baeri Brandt♀×A. schrenckii Brandt♂

The growth performance, body composition, fillet fatty acid content, serum hepatic enzymes and postprandial changes in serum lipid metabolism of hybrid sturgeon (70.8 ± 0.5 g) were investigated to determine the effects of total replacement of 80 g kg^‐1 fish oil (diet A) with linseed oil (diet B) and soybean oil (diet C), respectively. No significant differences in weight gain rate and specific growth ratio were observed among all fish groups (p > .05). Diet A fish group had the highest, but diet B fish group had the lowest feed intake (p < .05). Feed efficiency of fish fed diet A was significantly lower than the other two fish groups (p < .05). Fish fed diet A had the lowest fillet and liver lipid contents (p < .05). Serum lactate dehydrogenase, alanine aminotransferase and aspartate aminotransferase activities of fish fed diet A were significantly lower than those of fish fed diets B and C (p < .05). The contents of linoleic acid (C18:2n6) and linolenic acid (C18:3n3) in fillets showed a significantly positive linear correlation with the diets. Serum glucose and non‐esterified fatty acid were just affected by the time point (p < .05). The lipid source, time point and interaction of both factors had significant effects on serum triglyceride, high‐density lipoprotein cholesterol and low‐density lipoprotein cholesterol (p < .05). Serum total cholesterol was only affected by interaction of time point and lipid source (p < .05), and ketone body was not affected by lipid source, time point or interaction of both factors (p > .05). In summary, total replacement of fish oil with linseed oil or soybean oil had no significant adverse effects on hybrid sturgeon growth during 84‐day period, and linoleic acid and linolenic acid in fillet were modified by dietary treatments.     

Modulation of growth,immunity and antioxidant‐related gene expressions in the liver and intestine of juvenile Sillago sihama by dietary vitamin C

An 8‐week feeding experiment was implemented to study the effect of dietary vitamin C (VC) on growth, immunity, oxidation resistance and relevant gene expressions in juvenile S. sihama (2.33 ± 0.02 g). Fish were fed six diets containing VC level of 5, 16, 27, 65, 122 and 233 mg/kg, respectively. VC‐deficient signs including ascites syndrome, skin decay and haemorrhage were found in the fish fed basal diets. Appropriate VC supplement in diets (a) increased weight gain (WG), specific growth rate (SGR), feed intake (FI) and protein efficiency ratio (PER); (b) increased contents of hepatic total immunoglobulin (IgM), complement 3 (C3) and complement 4 (C4) and activity of alkaline phosphatase (AKP); (c) increased hepatic and intestinal copper–zinc superoxide dismutase (CuZnSOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione S‐transferase (GST) activities, but decreased hepatic MDA content; and (d) up‐regulated gene expressions of NF‐E2‐related factor 2 (Nrf2), Kelch‐like ECH‐associated protein 1 (Keap1), CAT, GST, GPx, glutathione reductase (GR), CuZnSOD and manganese superoxide dismutase (MnSOD) in liver and intestine. These results indicated that dietary VC significantly influenced the growth, immunity and oxidation resistance at enzymatic and genic levels. Based on regression analysis for PER, IgM, intestinal CuZnSOD and hepatic VC concentration, the optimal requirement of dietary VC in juvenile S. sihama was estimated to be 98.33, 139.03, 104.23 and 143.69 mg/kg.     

Low levels of Aflatoxin B1 could cause mortalities in juvenile hybrid sturgeon,Acipenser ruthenus ♂×A. baeri♀

Toxicity of aflatoxin B₁ (AFB₁) was investigated in juvenile hybrid sturgeon Acipenser ruthenus ♂ × A. baeri♀, an important coldwater finfish farmed in China and other countries. Seven experimental diets (Diet A–G) containing different levels of AFB₁ (0, 1, 5, 10, 20, 40 and 80 μg kg^?1 diet) were fed to juvenile sturgeon weighing 10.53 ± 0.17 g kg^?1 to determine its effect on survival, growth, feed consumption, hematocrit, liver histology as well as muscular and hepatic toxin accumulation. The experiment lasted for 35 days and was conducted in two periods of 25 and 10 days each. No external changes or unusual behaviour was observed in the fish fed diets with AFB₁. Mortality was observed in fish fed with highest levels of AFB₁ (80 μg kg^?1– Diet G) from day 12 onwards. After 25 days, fish fed the diet of 80 μg AFB₁ kg^?1 showed significant lower survival (50 ± 5.77%) followed by those fed 40 μg AFB₁ kg^?1 diet (80 ± 5.77%) and 20 μg AFB₁ kg^?1 diet (86.66 ± 3.33%). No significant difference was observed in specific growth rate (SGR) or hepatosomatic index (HSI) between groups. Hematocrit was significantly higher in the fish fed the diet of highest AFB₁. The fish were weighed at day 25 in some treatments (Diets F and G) because of high mortality. However, feeding was continued for another 10 days to observe mortality or behavioural changes if any in the other groups. After 35 days, survival in the fish fed Diet F (40 μg AFB₁ kg^?1) was 40% and those fed Diet E (20 μg AFB₁ kg^?1) was 36.2%. Significant histopathological changes including nuclear hypertrophy, hyperchromasia, extensive biliary hyperplasia, focal hepatocyte necrosis and presence of inflammatory cells were observed in the liver of fish fed high levels of aflatoxin (40 and 80 μg kg^?1). AFB₁ accumulation in fish muscle and liver increased with increased dietary AFB₁ levels. It could be confirmed that 10 μg AFB₁ kg^?1 diet was the maximum allowable level in hybrid sturgeon diet.