尼罗罗非鱼与萨罗罗非鱼及其正反杂交后代的微卫星分析

微卫星（microsatellite）技术是近年来广泛应用的DNA标记,具有重复性好、较易操作和共显性的特点。在硬骨鱼类中表现出高度多态性,已在家系鉴定、遗传作图、群体遗传分析、育种以及系统发生研究等多个领域得到应用。尼罗罗非鱼（Oreochromis niloticus）是我国主要淡水养殖鱼类,生长快,耐盐能力一般;萨罗罗非鱼（Sarotherodon melanotheron）耐盐能力强,适合咸淡水养殖,但生长较慢。[第一段]     

尼罗罗非鱼与萨罗罗非鱼正反交鱼自繁后代F2耐盐性、生长性能及亲本对杂种优势贡献力的评估

为选育耐盐和生长兼优的罗非鱼,以原始亲本尼罗罗非鱼(Oreochromis niloticus)和萨罗罗非鱼(Sarotherodon melanotheron)为对照,正交鱼(尼罗罗非鱼♀×萨罗罗非鱼↑)自繁第二代(F2)和反交鱼(萨罗罗非鱼♀×尼罗罗非鱼↑)自繁第二代(F2)为试验对象,观察和比较他们在盐度为0、15、20及25时的耐盐性和生长性能,并估算杂种优势及亲本的贡献力.结果如下:(1)盐度20～25是正、反交鱼的适宜生长盐度,他们在这一盐度范围里的日均增重率为尼罗罗非鱼在盐度0下的75%左右、萨罗罗非鱼在盐度25下的4倍左右,表明杂交后代的生长速度比较接近尼罗罗非鱼,远优于萨罗罗非鱼.在盐度25下,正交鱼比反交鱼生长快7%左右.(2)在盐度15与20下,生长速度和体重变异系数的杂种优势较明显,未发现不同盐度会导致正反交鱼成活率有显著变化.(3)不同亲本鱼类的强势性状,如尼罗罗非鱼的快速生长、萨罗罗非鱼的高耐盐性,对杂交后代的不同性状在不同盐度中有不同的贡献力.运用F1≤a1P1 a2P2所作的估算表明,对杂交后代在15～25盐度下的生长率,尼罗罗非鱼的杂种优势贡献力比萨罗罗非鱼的大3～4倍;对杂交后代在25盐度下的耐盐性,尼罗罗非鱼的贡献力几乎为零,而萨罗罗非鱼的贡献力几乎为100%.     

4种遗传型罗非鱼的耐盐慢性驯化表现

在实验室小水体中,对尼罗罗非鱼(Oreochromis niloticus,新吉富)、萨罗罗非鱼(Sarothrodon melanotheron)以及它们的杂交F1和F2("吉丽"罗非鱼)进行盐度梯度实验,观察其耐盐能力。采用4(遗传型)×4(水平或重复)随机区组设计。对照组全程维持淡水养殖;实验组起始盐度为0,盐度每天提高8直至实验鱼100%死亡。在水温保持(27.2±1.3)℃、正常投饵情况下,萨罗罗非鱼、尼罗罗非鱼、杂交F1及F2的50%死亡盐度平均值(MLS)分别为125.78±1.66、54.22±2.51、77.33±1.89和73.73±1.32;萨罗罗非鱼的累计存活率(CS)和MLS值均显著高于尼罗罗非鱼、杂交F1和F2(P<0.05);杂交F1和F2的CS值及MLS值均显著高于尼罗(P<0.05);F2和F1之间CS值和MLS值差异不显著(P>0.05)。     

大口黑鲈北方亚种和佛罗里达亚种及其杂交子代的遗传分析

用18对微卫星引物对大口黑鲈北方亚种(Micropterus salmoides salmoides,N)、佛罗里达亚种(M.salmoides floridanus,F)及其正交子代(N♀×F♂)和反交子代(F♀×N♂)进行遗传多样性和遗传结构分析。结果表明,18对引物扩增出的等位基因数为2~8个,平均等位基因数为5.0。检测到6对(Jzl48、Jzl68、Jzl84、MiSaTPW76、Msal21、Mdo6和Mdo7)亚种间特异性引物,其中有2对引物(Jzl48和Mdo7)可以用来鉴别大口黑鲈北方亚种、佛罗里达亚种和其杂交子代。平均有效等位基因数、平均期望杂合度和平均多态信息含量均为杂交组合F♀×N♂最高,分别为3.199 7,0.638 9和0.570 6,平均观测杂合度为杂交组合N♀×F♂最高(0.848 8)。对亲代与杂交子代间的遗传分化分析表明,正反交子代均与北方亚种的遗传分化最小(0.092和0.119 6)。基于Nei’s遗传距离构建的UPGMA系统进化树显示正交子代N♀×F♂与母本N聚为一支,反交子代F♀×N♂与父本N聚为一支。     

Growth,body composition,ammonia tolerance and hepatopancreas histology of white shrimp Litopenaeus vannamei fed diets containing different carbohydrate sources at low salinity

Juvenile Litopenaeus vannamei farmed at 3.0 psu were fed five diets containing glucose, sucrose, wheat starch, corn starch or potato starch as the carbohydrate (CBH) source. Shrimp were fed for 50 days to explore the effect of dietary CBH source on growth, body composition and ammonia tolerance. The specific growth rate of body length of shrimp fed glucose was the highest and significantly higher than those fed potato starch. The survival rate of shrimp fed glucose was 89.44%, and it was the highest and significantly higher than those fed wheat starch. Whole shrimp body crude protein and lipid of the corn starch group were 140.2 g kg^?1 and 10.1 g kg^?1 respectively. And they were significantly higher than those fed wheat starch. Shrimp fed potato starch had higher hepatopancreas and muscle glycogen. Shrimp fed sucrose had higher glucose‐6‐phosphate dehydrogenase and lower pyruvate kinase activities (P < 0.05). Besides, shrimp fed starch produced more B cells in hepatopancreas tubules than those fed glucose or sucrose. Shrimp fed different sources of CBH differed in the number of R cells. After 96‐h of ammonia nitrogen challenge, the survival rate of the treatments from high to low in turn was glucose, wheat starch, corn starch, sucrose and potato starch, and no significant differences were observed among all treatments. Based on shrimp growth and the economic problems of practical production, we recommend wheat starch as CBH source in practical diets for L. vannamei farmed at low salinities.     

Comparison of salinity tolerance and osmoregulation in two closely related species of blennies from different habitats

This study compares osmoregulatory capabilities of two closely related blennies inhabiting different habitats: Salaria pavo which inhabits marine rocky coasts in the Mediterranean and the eastern coasts of the Atlantic Ocean, and Salaria fluviatilis which inhabits freshwater habitats around the Mediterranean, both in rivers connected to the sea and in lakes that for a long time have been separated from the sea. Fishes for this study were collected in Lake Kinneret (Sea of Galilee) (S. fluviatilis) and Habonim beach, Israel, East Mediterranean coast (S. pavo). Both species tolerated salinities of 0, 40 and 100% seawater (0, 14.4 and 36 ppt sea salt respectively). Acclimation for a period of three months to freshwater (S. pavo) and seawater (S. fluviatilis) did not affect survival rates. After acclimation, total body water content of S. fluviatilis remained unaffected by experimental salinities but increased in S. pavo as salinity decreased. The hematocrit of both species increased as salinity increased. Plasma osmolality and Na⁺ concentrations were unaffected by experimental acclimation salinities in S. fluviatilis, but significantly decreased in S. pavo as salinity decreased. The results indicate that S. fluviatilis is able to osmoregulate both in fresh and seawater. S. pavo, living in seawater, while able to tolerate freshwater environment, fails to maintain constant extracellular fluid concentration. It is suggested that S. pavo, or its marine ancestor, has euryhaline capabilities that enabled it to invade freshwater habitats. S. fluviatilis, as the evolutionary result of this invasion, is thus better adapted to freshwater environment, but did not lose its ability to live in seawater.     

Temperature and salinity tolerances of yellowfin sea bream, Acanthopagrus latus, at different salinity and temperature levels

Abstract Yellowfin sea bream Acanthopagrus latus (Houttuyn) (0.98 ± 0.27 g) were examined for temperature and salinity tolerances at nine different environmental regimes (0.3, 15 and 33 ppt combined with 10, 25 and 32 °C), in which the fish were subjected to gradual and sudden changes in temperature and salinity respectively. The 50% CTMax (critical thermal maximum) and the UILT (upper incipient lethal temperature) were in the range of 34.8–38.2 °C and 32.8–36.4 °C respectively. The 50% CTMin (critical thermal minimum) and the LILT (lower incipient lethal temperature) were in the ranges of 4.9–9.4 °C and 7.1–17.6 °C respectively. The 50% CSMax (critical salinity maximum) and the UILS (upper incipient lethal salinity) were in the ranges of 54–69‰ and 9–44‰, respectively. The fish at lower temperature (10 °C) and salinity of 33‰ tolerated temperature as low as 6 °C when the temperature was decreased gradually. The fish at 33 and 15‰ and lower temperature (10 °C) tolerated salinity as high as 66–67‰, whereas those at freshwater salinity level (0.3‰) and 32 °C tolerated salinity to 50‰ when salinity was increased gradually. The relationships among UILT, LILT, 50% CTMax, 50% CTMin, UILS, 50% CSMax, salinity and temperature are given.     

尼罗罗非鱼与萨罗罗非鱼正反杂交后代耐盐性能的杂种优势及其与遗传的相关性的SSR 分析

对尼罗罗非鱼(Oreochromis niloticus)♀×萨罗罗非鱼(Sarotherodon melanotheron)♂(F2)、萨罗罗非鱼♀×尼罗罗非鱼♂(F2)、尼罗罗非鱼、萨罗罗非鱼4个遗传型群体的耐盐性实验表明:(1)4个遗传型群体的平均成活时间(MST)、50%成活时间(ST50)以及96 h半数致死浓度(MLS-96)由高到低依次为:萨罗、萨罗×尼罗(F2)、尼罗×萨罗(F2)、尼罗,死亡率与盐度具有显著的回归关系(P<0.05);(2)两个杂交种超越尼罗的超亲杂种优势值(HN)表现为正值,超越萨罗的超亲杂种优势值(HS)表现为负值,说明它们的耐盐力都超过了尼罗,但都未超过萨罗;(3)尼罗×萨罗(F2)的平均杂种优势值(HM)表现为负值,萨罗×尼罗(F3)的HM除其MLS-96表现为负值,其MST和ST50均为正值,说明萨罗×尼罗(F2)的耐盐性能略优于尼罗×萨罗(F2).对尼罗×萨罗(F2)、萨罗×尼罗(F2)、尼罗×萨罗(F1)、萨罗×尼罗(F1)、尼罗、萨罗6个遗传型群体的SSR分析发现:(1)有效等位基因数(Ne)、平均遗传杂合度(He)及多态信息含量(PIC)3项指标一致,表明F2遗传多样性比亲本增强2/3左右,这与杂交种的基因重组有关:F2又比F1增强1/10左右,初步认为这与F2的遗传分化有关;(2)引物Os-64和Os-75仅在尼罗、尼罗×萨罗(F1)及尼罗×萨罗(F2)中扩增出条带,表现出强烈的尼罗母系遗传:引物Os-25和IGF仅在萨罗、萨罗×尼罗(F1)及萨罗×尼罗(F2)中扩增出条带,表现出强烈的萨罗母系遗传,这4条引物可作为判别杂交鱼母本来源的遗传标记.     

不同盐度下尼罗罗非鱼、萨罗罗非鱼和以色列红罗非鱼幼鱼生长、成活率及肥满系数的差异

用1m×1m×1m的实验网箱,在盐度分别为0、10、20及30的水体中进行尼罗罗非鱼(Oreochromisniloticus)、萨罗罗非鱼(Sarotherodonmelanotheron)和以色列红罗非鱼(Israeliredtilapia)幼鱼的养殖实验。结果表明:(1)盐度、鱼的种类及盐度-鱼类的交互作用都对3种罗非鱼的生长及体重变异系数有显著影响(P<0 05);3种罗非鱼的成活率和肥满系数只受鱼的种类影响(P<0 05);盐度、盐度-鱼类的交互作用对其都无显著影响。(2)生长和盐度之间的回归关系在尼罗罗非鱼和以色列红罗非鱼较显著,在萨罗罗非鱼显著性较差。(3)尼罗罗非鱼和以色列红罗非鱼的生长随着盐度的降低而加快,萨罗罗非鱼的生长随着盐度的降低而减慢。在3种鱼中,盐度在6 9以下时,尼罗罗非鱼生长最快;盐度7 4～28 7时,以色列红罗非鱼生长最快;盐度高于29 0时,萨罗罗非鱼生长最快。(4)在实验的4种盐度下,萨罗罗非鱼的成活率都高于以色列红罗非鱼和尼罗罗非鱼,而除了在淡水中外,以色列红罗非鱼的成活率又都高于尼罗罗非鱼;萨罗罗非鱼的肥满系数都显著高于尼罗罗非鱼和以色列红罗非鱼,而以色列红罗非鱼的肥满系数和尼罗罗非鱼的没有显著差异。     

Osmoregulation,growth and moulting cycles of the giant freshwater prawn (Macrobrachium rosenbergii) at different salinities

The giant freshwater prawn, Macrobrachium rosenbergii, is a species with a high commercial value in aquaculture. Two experiments were performed to determine the effects of salinities on the osmoregulation, growth and molting cycles of M. rosenbergii during growout. The first experiment was designed to determine whether these animals are capable of adapting to the changes in salinity seen in salinity intrusions in tropical deltas, with an incremental increase in salinity of 3‰ per day from 0‰ to 30‰ Haemolymph osmolality was rapidly regulated up to salinities of 15‰ , whereas animals conformed at higher salinities. The second experiment determined the growth, moulting cycle, osmolality, muscle water content and mortality during a 4‐month experiment at 0‰, 15‰ or 25‰ salinity. The weight gains in 0‰ and 15‰ were not significantly different and were comparable to the growth rates achieved in production farms with body mass increases of 2.6 and 2.3‐fold their initial body mass, respectively, after 4 months. The 25‰ group suffered from low growth, high mortality and a significantly lower moulting frequency. These data show that this species can be reared in brackish water up to 15‰, allowing for farming in the large areas impacted by salt water intrusions in tropical deltas.     

Effect of replacing soybean meal with canola meal on growth,feed utilization and haematological indices of juvenile hybrid tilapia,Oreochromis niloticus×Oreochromis aureus

An 8‐week feeding trial was conducted to evaluate the effects of replacing soybean meal (SBM) with canola meal (CM) on growth, feed utilization, body composition and haematological indices of juvenile hybrid tilapia (Oreochromis niloticus×Oreochromis aureus). Six isonitrogenous diets containing graded levels of CM (0, 95, 190, 285, 380 and 634 g kg^?1 of diet corresponding to 0%, 15%, 30%, 45%, 60% and 100%, respectively, of protein from SBM) to replace SBM on an equal protein basis were fed to triplicate groups of juvenile fish (initial weight=6.3 g). The results indicated that up to 30% of SBM could be replaced by CM without causing a significant reduction in growth performance. Fish fed with diets in which CM replaced over 45% of SBM had a significantly lower protein efficiency ratio and a significantly higher feed conversion ratio than fish fed with other diets. The apparent digestibility coefficients (ADC) of dry matter, protein and phosphorus were lowest for fish fed the CM100 diet. Significant differences in haemoglobin, haematocrit and white blood cell concentration were found in fish fed diets with different CM levels. It is concluded that up to 19.02% CM can be used to replace 30% of SBM in diets for juvenile hybrid tilapia without compromising growth, feed conversion and protein utilization.     

Growth and osmoregulation in Atlantic salmon (Salmo salar ) smolts in response to different feeding frequencies and salinities

Groups of Atlantic salmon (Salmo salar) yearling smolts were reared in duplicate tanks supplied with freshwater or seawater, and subjected to different feeding frequencies, 100% (fed every day), 50% (fed every other day), 25% (fed every forth day) and 0% (starved), from 26 May to 26 July. After 8 weeks, all the groups were re‐fed in excess for 6 weeks. Fish were maintained on their respective a priori salinity treatments during the 6‐week follow‐up period. Starvation for a period of 8 weeks in freshwater resulted in a loss of hypo‐osmoregulatory ability when smolts were challenged with seawater and unfed smolts maintained in freshwater were unable to adapt to seawater in mid‐July. Ration levels influenced the growth rate and body size significantly. The overall growth rate was higher in freshwater than at corresponding rations in seawater. Partial compensatory growth was observed in the 0 and 25% groups following re‐feeding. Branchial Na⁺,K⁺‐ATPase (NKA) activity decreased rapidly in unfed smolts in freshwater and was the lowest in the starved group, whereas an initial increase was observed in those groups reared in seawater. After re‐feeding NKA activity differences decreased between the former feeding groups. Our results suggest that nutritional factors and/or energy levels are critical for the maintenance of hydro‐mineral balance of salmon smolts.     

Short-term salinity tolerance of northern pike, Esox lucius, fry, related to temperature and size

Abstract  The short-term tolerances of northern pike, Esox lucius L., fry reared in a freshwater hatchery, to salinity were examined in the laboratory. Survival of two size groups of pike fry (mean length 21 ± 2 mm SD and 37 ± 4 mm SD) was examined over 72- to 96-h periods at 9–14 ppt salinity in combination with temperatures of 10, 14 and 18 °C. A parametric survival model found a significant correlation between survival of pike fry and temperature and salinity, respectively. L _C50 values after 72 h were between 11.2 and 12.2 ppt, being lowest at 10 °C. Pike fry did not survive more than 13 ppt. Mortality at 12 ppt was significantly faster at 18 °C than 10 or 14 °C. Moreover, mortality was higher and faster for large than for small pike fry at 12 ppt and 14 °C. These results imply that pike raised in fresh water can survive stocking into brackish waters below 11 ppt at least for a short time.     

Growth performance,osmoregulatory and metabolic modifications in red porgy fry,Pagrus pagrus,under different environmental salinities and stocking densities

The effects of two different environmental salinities [brackish water (BW), 12‰; sea water (SW), 39‰] and initial stock densities [low (LD), 1.0 g L^?1; high (HD), 2.0 g L^?1] on growth, osmoregulation, stress and energy metabolism of the fry Pagrus pagrus were investigated over a period of 45 days. Pagrus pagrus (n=80, 5.51 ± 0.25 g mean initial body weight) were randomly divided in eight groups. Growth, weight gain and specific growth rate increased in BW‐acclimated fish compared with SW‐acclimated fish. No differences were observed between the two stock densities tested at either environmental salinity. Plasma osmolality was lowest in BW‐acclimated specimens, but the stock density had no effect on this parameter. Branchial Na⁺,K⁺‐ATPase activity was positively correlated with environmental salinity, but unaltered at the renal level. Plasmatic parameters were enhanced by salinity and stocking conditions. At the hepatic level, triglyceride values were enhanced in BW‐acclimated fish maintained at LD. Muscle metabolites (glycogen, glucose and lactate) increased in BW‐ compared with SW‐acclimated fish; stock density had no influence. Our data suggest that changes in metabolic parameters could be correlated with the higher growth rates observed in P. pagrus acclimated to BW, while no significant effects due to the stocking density used were observed.     

Growth performance,haematological parameters,antioxidant status and salinity stress tolerance of juvenile Pacific white shrimp (Litopenaeus vannamei) fed different levels of dietary myo‐inositol

A 10‐week growth trial was run to evaluate effects of myo‐inositol (MI) on growth performance, haematological parameters, antioxidative capacity and salinity stress tolerance of Litopenaeus vannamei. Six practical diets supplemented with graded levels of MI (designated as MI0, MI600, MI1200, MI2400, MI 3600 and MI4800 for 448.8, 974.2, 1568.0, 2810.6, 3835.5 and 4893.6 mg/kg diet, respectively) were fed to six replicate groups of L. vannamei (mean initial body weight 0.63 ± 0.00 g). The results showed that significant increment of growth performance was observed in shrimp fed MI600 diet than those fed MI1200 diet. Lipid concentration in whole body of the shrimp fed MI600 diet was significantly increased. Shrimp fed MI0 diet had lower total protein (TP) as compared to shrimp fed the MI‐supplemented diets (except MI4800 diet). In general, lower activities of antioxidant enzymes and higher malondialdehyde (MDA) content in haemolymph and hepatopancreas were recorded in shrimp fed MI0 diet, compared to those fed the MI‐supplemented diets. Reduced survival after 7‐h salinity stress was present in shrimp fed MI0 diet as compared to those fed MI4800 diet. Dietary MI requirement for glutathione peroxidase activity of L. vannamei was 2705 mg/kg diet.     

盐度对奥尼罗非鱼仔稚鱼生长、存活及其消化酶活力的影响

研究了不同盐度对奥尼罗非鱼仔、稚鱼生长、存活和消化酶活力的影响。试验设计2、4、6、8、10和纯净淡水0（对照组）共6个盐度梯度组,仔鱼的初始体重0.0080.001 g。结果表明,不同盐度对奥尼罗非鱼仔、稚鱼生长有显著差异（P0.05）,盐度4和6时,仔、稚鱼的生长速度和绝对增重率最高,分别为0.08 cmd-1和0.033 gd-1;盐度10时较低,只有0.06 cmd-1和0.023 gd-1;仔、稚鱼的肥满度介于3.43~3.81,各组之间差异不显著（P0.05）;盐度0~10时,各组存活率介于92.3%~94.7%,幼鱼所占比例介于66.0%~67.3%,各组之间差异不显著（P0.05）;胃蛋白酶和胰蛋白酶活力随盐度升高而降低,均在盐度0时活力最高,10时最低;脂肪酶和淀粉酶活力在盐度4时达到最高,10时活力最低。根据试验结果,奥尼罗非鱼仔、稚鱼培育的盐度最好控制在4.6左右。     

Growth and proximate composition of the Chaetoceros calcitrans f. pumilus under different temperature, salinity and carbon dioxide levels

The marine diatom Chaetoceros calcitrans f. pumilus has been examined for its potential source as live feed in aquaculture. The present study investigated effects of temperature (20, 25 and 30 °C), salinity (25 and 35) and carbon dioxide addition (air+CO₂) on the growth and proximate composition of C. calcitrans under laboratory conditions. The growth and biomass of C. calcitrans were primarily affected by carbon dioxide addition, and to a lesser extent by temperature and salinity. In general, lipid and carbohydrate contents were higher at lower temperatures (20 and 25 °C), while the protein content was unaffected. Carbon dioxide addition increased protein, while lowering carbohydrates, but had no effect on lipid content. Carbohydrates were increased while lipids and protein decreased at the highest salinity (35±0.9). These results should be taken into consideration when evaluating the dietary value of this micro alga for aquaculture.     

Larval development and salinity tolerance of Japanese flounder (Paralichthys olivaceus) from hatching to juvenile settlement

Salinity tolerance and growth of Japanese flounder Paralichthys olivaceus at different developmental stages were evaluated, including newly hatched larvae (nhl), yolk sac larvae (ysl), oil droplet larvae (odl), post oil droplet larvae (podl), premetamorphic larvae (preml) and prometamorphic larvae (proml), at 11 salinities from 5 to 55 g L^?1 for 96 h. The ontogenesis during the early life of P. olivaceus was investigated under hatchery salinity 35 g L^?1. The results showed that suitable salinities for nhl, ysl, odl, podl, preml and proml larvae were 10 to 25 g L^?1, 10 to 30 g L^?1, 20 to 30 g L^?1, 30 g L^?1, 10 to 30 g L^?1, 15 g L^?1, respectively, demonstrating an ontogenetic variation of salinity tolerance. The salinity tolerance of nhl, ysl, preml was higher than that of odl, podl and proml. The ysl and preml larvae displayed wide salinity tolerances. The present findings demonstrate that the suitable salinity for larviculture of P. olivaceus is 20–25 g L^?1 before the depletion of oil droplet; after that, higher salinity (30 g L^?1) should be ensured for the post‐oil droplet larvae; the premetamorphic larvae can be cultured at a wide salinity range (10–30 g L^?1), and the metamorphosed larvae should be reared at salinity about 15 g L^?1.     

吉富品系尼罗罗非鱼选育系F6、F7和F8当年鱼生长对比研究

罗非鱼是世界性养殖鱼类之一,对罗非鱼进行遗传改良研究具有重要意义。在流水池饲养环境里,对选育系F6、F7和F8 3代的当年尼罗罗非鱼Oreochromis niloticus生长性能进行同池比较。经过114 d的饲养,F6、F7和F8世代鱼体重的平均生长速度分别为1.07、1.12和1.47 gd-1,呈逐代增大的趋势。偏差校正后,F6、F7和F8 3代鱼增重率分别为1.13、1.18和1.36 gd-1。F8的体重变异系数比F7降低了6.29%,比F6降低了7.95%。F8代的生长方程是y=40.313x－61.670（R2=0.8783,R=0.9513）。