Lactobacillus spp. bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae: Effects on growth performance and digestive enzyme activities

In this study, the influence of commercial probiotic, Lactobacillus spp., supplementation was investigated on growth parameters and digestive enzyme activities in gilthead sea bream, Sparus aurata, during larval development. All experiments were triplicated and designed in three different administrations of probiotic from 3 days after hatching (DAH) concurrently with starting of exogenous feeding. In the first group, probiotic was added to live food (rotifer and Artemia). In the second group, probiotic was supplemented directly to both live food and water. In the third group, probiotic was added directly to water. Also, no probiotic treatment was maintained in control group. Total bacterial counts among probiotic probiotic-supplemented groups were significantly different from total bacterial counts in controls in water and digestive tract of larvae (p < 0.05). The mean of total bacterial counts in control was approximately 4 × 10⁴-fold increased from the experimental groups in the sea water (p < 0.05). Besides, mean digestive enzyme activities of all probiotics treatment groups were significantly different (p < 0.05) with that of the control. Except probiotic water supplementation group, in all treatments, the specific activities of pancreatic and intestinal enzymes were significantly higher (p < 0.05) in larvae to which probiotic had been supplemented by live food and live food with water. Also, S. aurata larvae that had probiotic administered by live food with water demonstrated significant (p < 0.05) increases in both survival (13–105% higher) and specific growth rate (2–9% higher) as compared to controls. As a result, supplementation of probiotic to directly tank water could not significantly increase growth parameters and digestive enzyme activities and therefore, administration of probiotics by this method would not be effective in terms of husbandry parameters and nutritional condition.     

Heritability of skeleton abnormalities (lordosis, lack of operculum) in gilthead seabream (Sparus aurata) supported by microsatellite family data

Body abnormalities constitute an important problem to fish aquaculture. Pigmentation, scale and skeleton abnormalities have been reported in different cultured fish species. Environmental and genetic factors or their interaction have been suggested to explain their origin. Gilthead seabream (Sparus aurata) is one of the most important European cultured species. Lordosis and lack of operculum are common abnormalities in this species, and can reach frequency of 80% or more of commercial batches. In spite of an important body of data suggesting environmental factors related with their origin, scarce and not conclusive genetic data have been reported to date. In our study, a large number of families originating from the usual production process of a commercial farm were used to estimate the heritabilities of both characters. Two independent experiments were carried out for each abnormality. Nine hundred and ninety four (5.6% lordotics; 157 full-sib families; 6.3 offspring/family) and 808 (7.9% lacking operculum; 83 families; 9.7 offspring/family) individuals were used for lordosis and lacking operculum heritability estimations, respectively. The results obtained adjusting an animal model indicated non significant heritabilities for both characters (0.021 (s.e. 0.019) and 0.032 (s.e. 0.023) for lordosis and lack of operculum, respectively). The corresponding values when a threshold model was used, though higher (0.152 and 0.203, respectively), evidenced large standard errors (0.119 and 0.146, respectively) suggesting h² = 0 as the most confident hypothesis (P = 0.838 and 0.766, respectively). A non-parametric permutation test was also applied to evaluate if more related individuals had a higher phenotypic resemblance. The results obtained suggested only a slight familiar association (P < 0.05) when comparing individuals lacking operculum, but neither between lordotics nor between normal ones. These results suggest that most phenotypic variation observed for lordosis and lack of operculum in gilthead seabream is due to environmental factors.     

Genetic comparison of wild and cultivated European populations of the gilthead sea bream (Sparus aurata)

This study represents the first large-scale population genetic analysis of the marine fish gilthead sea bream (Sparus aurata), one of the most significant species in the South European aquaculture. Six wild and five cultivated sample sets covering the South Atlantic and Mediterranean European area have been screened for allozyme, microsatellite and mitochondrial DNA (mtDNA) variation. Microsatellites showed higher levels of polymorphism than allozymes. The low variability of mtDNA offered no basis for population differentiation.

The results reveal levels of variability for S. aurata above those from other sparids. Cultivated populations show a slight decrease of variability related to the wild ones, but not sufficient to document inbreeding depression effects, thus suggesting a fairly proper management. Wild populations reveal a slight degree of differentiation more pronounced with microsatellites than with allozymes, but not apparently associated with geographic or oceanographic factors. The cultivated populations seem to be highly divergent as a result of genetic drift caused by different factors pertaining to their respective histories. With both markers, the two cultivated Spanish sample sets are the most divergent. The high differentiation between cultivated and wild populations from the same area might indicate no evidence for significant genetic flow between them.

This study provides an insight into the population structure of S. aurata, although more questions have arisen that need to be solved. This can be achieved by further screening of small-scaled targeted sample sets in the studied area.     

n-3高不饱和脂肪酸对黑鲷幼鱼生长及脂肪代谢的影响

研究了饲料中不同水平n-3 HUFA（0.79%,0.83%,0.85%,0.88%,0.92%,0.94%;DHA/EPA=2.8/1）对黑鲷幼鱼生长及脂肪代谢的影响。结果显示：（1）黑鲷幼鱼肝体指数（HSI）及腹脂率（IPF ratio）随饲料中n-3 HUFA含量的增加而减小,且于0.92%和0.94%组时显著低于其他各组;脂肪细胞直径呈减小趋势,其中0.94%组显著小于0.85%组;肌肉脂肪含量受n-3 HUFA的影响显著,于0.88%组时达到最低。各组全鱼水份和脂肪含量差异不显著（P>0.05）。肝脏、肌肉及腹腔脂肪组织饱和脂肪酸（∑SFA）和C16∶0含量均随饲料n-3 HUFA水平增加呈下降趋势,而∑n-3 HUFA呈显著上升趋势。各组织中DHA/EPA不受饲料脂肪酸组成的影响。以增重率为参考指标,二次回归分析结果表明,黑鲷幼鱼［（8.08±0.09） g］获得最佳增重时对饲料中n-3 HUFA的需要量为0.87% DM;（2）黑鲷幼鱼肝脏脂肪酸合成酶（FAS）活性及基因表达水平均在n-3 HUFA>0.92%时有显著下降（P<0.05）;腹腔脂肪激素敏感脂肪酶（HSL）活性及基因表达水平均随饲料中n-3 HUFA的添加呈升高趋势（P<0.05）,且高含量n-3 HUFA（0.94%）可使HSL活性增加近一倍。结果表明,饲料中n-3 HUFA通过同步调控脂肪合成与分解两个过程影响黑鲷幼鱼脂肪代谢。     

LED光谱对舌齿鲈幼鱼摄食、生长和能量分配的影响

在海水循环水养殖系统中,分别设置白光(λ_(400～780 nm))、红光(λ_(630 nm))、黄光(λ_(595 nm))、绿光(λ_(530 nm))、蓝光(λ_(455 nm))5组不同LED光谱处理组,研究5种不同LED光谱对舌齿鲈幼鱼[(29.91±0.39) g、(13.78±0.35) cm)]摄食、生长和能量分配的影响。结果显示,不同LED光谱对舌齿鲈幼鱼摄食、生长和能量分配具有显著性影响。红光谱下舌齿鲈幼鱼生长较好(41.09±5.70) g,蓝光谱下舌齿鲈幼鱼生长较差(36.02±4.18) g;蓝光谱下舌齿鲈幼鱼DGC(daily growth coefficient)显著低于其他各组,其他各组之间差异不显著;5组LED光谱处理组舌齿鲈幼鱼摄食率存在显著性差异,其中红光谱摄食率最高。类胰岛素生长因子IGF-1和IGF-2测定结果表明,白光谱下IGF-1的mRNA表达量最高;黄光谱下IGF-2的mRNA表达量最高。能量分配方面,红光谱下舌齿鲈幼鱼排粪能和代谢能较低,摄食能和生长能较高;蓝光谱下舌齿鲈幼鱼排粪能较高,摄食能和代谢能较低,生长能最低。对舌齿鲈幼鱼肌肉和肝脏RNA/DNA比值测定结果表明,蓝光谱下,舌齿鲈幼鱼肝脏RNA/DNA比值显著高于其他各组;蓝光谱和白光谱下舌齿鲈幼鱼肌肉中RNA/DNA比值存在显著性差异,其他各组之间不具有显著性差异。研究表明,红光谱下舌齿鲈幼鱼摄食、生长均表现较好,且红光谱下舌齿鲈幼鱼具有较高的摄食能和生长能;蓝光谱下舌齿鲈幼鱼摄食、生长表现较差,排粪能较高,生长能较低。因此,红光谱对舌齿鲈幼鱼摄食、生长具有积极影响;蓝光谱对舌齿鲈幼鱼摄食、生长具有不利影响。     

Abnormalities in the juvenile stage of sea bass (Dicentrarchus labrax L.) reared at different temperatures: types, prevalence and effect on growth

Osteological malformations are always considered an important problem in intensive aquaculture. This work studies the effect of rearing temperature on malformations in sea bass, Dicentrarchus labrax. To this end, two batches of the species were subjected to the following incubation/cultivation temperatures: 15 °C/natural and 19/19 °C, from fertilization until 190 days after hatching. The different malformations were studied in 1643 juveniles at 190 days and classified by categories. The Chi-square statistic was calculated to verify the relationship between the presence of anomalies and the application of different temperatures. The percentage of anomalies observed in individuals reared at a high temperature (19/19 °C) was 66.44%. In both temperature systems, opercular malformations were those which caused a greater delay in growth. The results found indicated that temperature played a very important role in the development of deformations, which may be of interest from the viewpoint of aquaculture.     

Self-feeding activity patterns in gilthead sea bream (Sparus aurata), red porgy (Pagrus pagrus) and their reciprocal hybrids

This study was conducted to monitor feeding activity of gilthead sea bream (Sa) and red porgy (Pp) and to investigate the hereditary effects on feeding in their hybrids (female Sa×male Pp, female Pp×male Sa), obtained after artificial crosses. All fish groups were held in 500-l tanks under ambient temperature (15–25°C) and light conditions (daylength from 9 h 30 min to 14 h 48 min). Fish (initial body weight 100–165 g) were fed by means of self-feeders, which were connected to a computer, recording fish feeding activity for a year. Results showed that there was a 24-h periodicity in all fish types over the experiment. In their daily feeding patterns, both parental species were diurnal: red porgies had a stable daily pattern over the experiment, while gilthead sea bream shifted their activity from a general daylight fluctuation in warm periods to an afternoon peak in cold periods. The Sa×Pp hybrids showed intermediate feeding patterns compared to their parents in warm periods and a pattern that was similar to the red porgy in cold periods, while the Pp×Sa hybrids had rather unchangeable daily feeding patterns, which were not related to the parental ones. The proportion of night feed demands increased in cold periods independent of fish group. The relationship between temperature and feed intake was species-dependent. Finally, this study indicated that feeding expression in hybrids was related to the combination of the parental genome, supporting our hypothesis for a hereditary influence on feeding activity in fish.     

In vivo and in vitro treatments against Sparicotyle chrysophrii (Monogenea: Microcotylidae) parasitizing the gills of gilthead sea bream (Sparus aurata L.)

The effect of in vitro and in vivo treatments against Sparicotyle chrysophrii, a microcotylid parasite of gilthead sea bream (Sparus aurata L.), was studied. In vitro chemical treatments were targeted to eggs, oncomiracidia and adults, and were tested both as disinfectants and therapeutics for infected animals. The compounds were: distilled water, formalin, limoseptic ®, hydrogen peroxide, chlorine, and praziquantel (PZQ). Larvae were sensitive to all the treatments, but adults were more resistant, as chlorine (60 ppm - 1 h), hydrogen peroxide (100 ppm - 30 min) and PZQ (50 ppm - 30 min) produced only 10% mortality. All adults were killed only with distilled water, limoseptic (0.1% - 5 min), formalin (300 ppm - 30 min), or hydrogen peroxide (200 ppm - 30 min). Eggs were the most resistant stage, as only 30 min in limoseptic (0.1% in distilled water) or in formalin (300 ppm) prevented hatching. PZQ was used in vivo either as a curative or preventive treatment. The highest dose tested (400 mg kg^− 1 BW; effective dose 116.3 mg kg^− 1 BW due to palatability problems leading to 45% reduction in host food intake) did not significantly decrease prevalence of infection when given for 6 consecutive days. A lower dose (200 mg kg^− 1 BW) (effective dose 158.1 mg kg^− 1 BW) was rejected to a lower degree and decreased the prevalence of infection from 90% to 40%. When a lower dose (40 mg kg^− 1 BW) was administered for longer periods (20 days), food intake was reduced slightly, but the infection did not decrease significantly. The oral intubation with PZQ (200 mg kg^− 1 BW) once a week for 4 weeks did not prevent the infection of fish by cohabitation. However, a significant reduction in the abundance of the parasite was registered. In view of the results, recommendations for fish treatment and disinfection of aquaculture facilities are discussed.     

Evaluation of three seaweeds Gracilaria bursa-pastoris, Ulva rigida and Gracilaria cornea as dietary ingredients in European sea bass (Dicentrarchus labrax) juveniles

The aim of this study was to evaluate the inclusion of three seaweeds Gracilaria bursa-pastoris (GP), Ulva rigida (UR) and Gracilaria cornea (GC) as dietary ingredients on the performance, nutrient utilisation and body composition of European sea bass juveniles. Six experimental diets were formulated to replace 5% (GP-5, UR-5, and GC-5 Diets) and 10% (GP-10, UR-10 and GC-10 Diets) fish protein hydrolysate (CPSP) by each of the three seaweeds. A control diet was used, without inclusion of any seaweed. Diets were fed to duplicate groups of 25 European sea bass (Dicentrarchus labrax) juveniles (IBW = 4.7 g) for 10 weeks. Growth performance was only significantly reduced (P < 0.05) in fish fed the GC-10 diet, whereas the feed conversion ratio increased significantly in those fish. The apparent digestibility coefficients of dry matter and lipid were significantly lower in fish fed diet GC-10 relative to those fed the control diet. Carcass composition was similar among treatments, although fish fed GC-10 exhibited significantly higher ash content.The results obtained in this study suggest that the inclusion of G. bursa-pastoris (GP) and U. rigida (UR), up to 10%, can be considered as very interesting ingredients in diets for sea bass juveniles, as no negative consequences on growth performance, nutrient utilization or body composition were observed. On the other hand, the inclusion of G. cornea (GC) should be limited to 5% of the diet.     

大口黑鲈ghrelin基因SNPs的筛选及与生长性状关联性分析

ghrelin是脊椎动物的一种脑肠肽,有促进摄食的功能,并能促进生长激素(GH)释放,参与能量平衡调控和糖类代谢。为探索ghrelin基因多态性与大口黑鲈生长性状的相关性,针对大口黑鲈ghrelin基因的启动子序列,实验采用直接测序法获得了2个SNPs位点:S1(A-642C)和S2(A-639C)。随机选取同批繁殖、同塘养殖的大口黑鲈采用Sna Pshot方法进行SNPs位点检测和分型。结果显示,实验群体在ghrelin基因2个SNPs位点上基本处于哈温平衡,S1和S2共组成5种双倍型(D1、D2、D3、D4和D5)。基因型与生长性状的相关性分析结果表明,S1位点AC型个体在体高与全长上显著高于CC型个体,S2位点AA型个体在头长上显著高于AC型个体。双倍型D1在体质量、全长和体高方面显著高于双倍型D3,在体质量、体宽和体高方面显著高于双倍型D4。本研究在大口黑鲈ghrelin基因启动子区域获得的与生长性状相关的SNPs标记,可为大口黑鲈分子标记辅助育种提供帮助。     

Effects of dietary amino acid profile on growth performance, key metabolic enzymes and somatotropic axis responsiveness of gilthead sea bream (Sparus aurata)

Juvenile gilthead sea bream were fed to visual satiety with isonitrogenous diets based on fish meal and different plant ingredients (33–35% replacement) supplemented with free amino acids to meet the desired indispensable amino acid (IAA) profile and dispensable amino acid (DAA) content. In diets M and WB, IAA profile and DAA content resemble that of the muscle or whole body, respectively. In diets MGlu and WBGlu, DAA content was increased by adding -glutamic acid (Glu) and thus the IAA/DAA ratio varied from 1.13 (diet M) to 0.80 (diet WBGlu). Growth rates were not significantly different among experimental groups, but feed conversion ratio and nitrogen retention were impaired by the decrease of dietary IAA/DAA ratio. Postprandial ammonia excretion increased with the increase of dietary DAA content irrespective of IAA profile. Conversely, hepatic activity of glutamate dehydrogenase (GDH) was lower in fish fed diet WBGlu than in fish fed diet M. Hepatic growth hormone (GH) binding was not significantly affected by the dietary treatment, but circulating levels of insulin-like growth factor-I (IGF-I) and GH were, respectively, down- and up-regulated in fish fed diet WBGlu, which suggests some defect in the transmission of GH receptor signal. Fat retention and hepatic activities of lipogenic enzymes (glucose-6-phosphate dehydrogenase, G6PD; malic enzyme, ME) were decreased in fish fed diet MGlu. Key metabolic enzymes of hepatic glycolysis (glucokinase, GK) and gluconeogenesis (phosphoenolpyruvate carboxykinase, PEPCK) were also altered in this group of fish. Since soybean meal concentration was highest in diet MGlu, results on lipid and carbohydrate metabolism can be primarily attributed to this component of the diet. In contrast, data on growth performance, ammonia excretion and GH axis mainly reflect changes in the dietary amino acid profile, which reveals that a muscle IAA profile and a high IAA/DAA ratio are important in feeds for gilthead sea bream.     

大口黑鲈幼鱼对饲料中锌的需求量

为确定大口黑鲈幼鱼对饲料锌的适宜需求量,以酪蛋白、明胶和鱼粉为主要蛋白质源,以ZnSO_4·H_2O为锌源制作半精制基础饲料,分别向基础饲料中添加0、25、50、100和200 mg/kg锌,制成5种含不同锌水平饲料(24.8、48.8、78.9、126.1和223.6 mg/kg干物质)(命名为Zn-25、Zn-49、Zn-79、Zn-126和Zn-224),投喂初始体质量为(10.99±0.07) g的大口黑鲈幼鱼8周。结果显示,饲料中补充25 mg/kg锌(Zn-49)显著提高了大口黑鲈增重率,降低了饲料系数,进一步提高锌的添加量后,各组增重率和饲料系数趋于稳定。当饲料锌含量为25～49 mg/kg时,T-SOD和CuZn-SOD活性增加,锌含量达到49 mg/kg后,其活性保持基本稳定,而AKP活性在Zn-79组最高。大口黑鲈全鱼和脊椎骨中的锌含量随饲料中锌含量的增加而上升,当饲料锌含量达到126 mg/kg (Zn-126)后,全鱼和脊椎骨中的锌含量不再显著增加,而全鱼铁、骨铁、骨锰含量和全鱼铁、锌沉积率则随饲料锌含量的增加而下降。研究表明,在半精制饲料中补充锌可以显著改善大口黑鲈的生长和饲料利用,提高血清免疫能力、全鱼锌和骨锌的沉积,以增重率、饲料系数、全鱼锌和骨锌为指标,基于折线模型确定大口黑鲈幼鱼对饲料中锌的需求量分别为45.5、44.6、121.8和130.5 mg/kg干物质。     

豆粕替代鱼粉对牛蛙生长、体组成、消化酶活力及肝脏生化指标的影响

为减少牛蛙饲料鱼粉使用量以降低养殖成本,研究了豆粕替代饲料中不同比例的鱼粉对牛蛙的生长性能、体组成、肠道消化酶活力和肝脏生化指标的影响。用豆粕分别替代0%、20%、40%、60%、80%和100%的鱼粉配制出6种等氮等能的实验饲料,每组饲料设3重复,养殖牛蛙60 d。结果显示,豆粕替代鱼粉对牛蛙摄食率、成活率、肝体指数、脏体指数、蛙体水分和灰分含量、蛙腿肌肉组成以及肝脏丙二醛含量的影响不显著;豆粕100%替代鱼粉显著降低牛蛙增重率、特定生长率、饲料系数、蛋白质效率、蛋白质累积率;随豆粕替代鱼粉比例的提高,牛蛙的肥满度和后腿指数呈下降趋势,替代100%鱼粉显著降低牛蛙肥满度,替代60%鱼粉显著降低牛蛙后腿指数;替代80%鱼粉显著降低牛蛙全体粗蛋白的含量;牛蛙全体粗脂肪含量随替代鱼粉比例的提高呈上升的趋势,替代100%鱼粉组粗脂肪含量最高;随着替代比例的提高,牛蛙肠道蛋白酶的活力显著降低,而肠道淀粉酶和脂肪酶活力显著提高。豆粕替代鱼粉显著影响肝脏超氧化物歧化酶和过氧化氢酶活力,其中超氧化物歧化酶活力最低值出现在替代80%鱼粉组,过氧化氢酶活力最低值出现在替代100%鱼粉组。基于以上实验结果,建议牛蛙饲料中豆粕可替代鱼粉的比例为60%~80%。     

仿刺参铁蛋白ferritin基因的序列分析及表达

通过构建仿刺参cDNA文库,获得铁蛋白全长cDNA序列。该基因序列全长792 bp,5′非翻译区长133 bp,开放阅读框长522 bp,编码173个氨基酸,3′UTR长137 bp;预测蛋白分子量为20 ku。5′UTR具有一个高度保守的铁离子应答元件。仿刺参ferritin氨基酸序列具备脊椎动物ferritin的亚铁氧化酶活性中心所特有的保守结构。该序列与海参的同源性最高,达84%,与其它无脊椎动物如:海星、鲍、牡蛎、海葵、线虫、小龙虾和果蝇的同源性为74%~34%;与脊椎动物ferritin重链亚基同源性高于轻链亚基。系统进化分析表明,仿刺参的ferritin与大部分无脊椎动物聚为一支。利用半定量RT-PCR检测,ferritin mRNA在仿刺参未受精卵、受精卵、多细胞期、囊胚期、原肠期、小耳状幼体、中耳状幼体、大耳状幼体、樽型幼体、五触手幼体、稚参11个发育阶段和幼参的体壁、体腔细胞、肠道和呼吸树中均表达。Quantitative real-time PCR结果显示, ferritin mRNA在未受精卵至原肠期表达量低,从小耳状幼体至稚参表达量显著增高;在幼参的不同组织中,ferritin mRNA在呼吸树中的表达量显著低于其他3种组织;幼参注射LPS后,4种组织中ferritin mRNA表达量与注射前无显著差异。     

鲤和草鱼IL17受体基因家族的全基因组识别、起源进化及表达分析

为了研究鲤科鱼类最具代表性的二个物种—鲤和草鱼IL17受体基因家族的起源进化,实验采用比较基因组学和生物信息学的方法,分别在鲤和草鱼基因组数据库进行序列比对和注释,然后对得到的基因进行结构域和系统发育学分析,最后在12个不同组织中进行基因表达分析研究。结果显示,在鲤和草鱼中分别注释得到9个和5个IL17受体基因家族成员。系统发育分析显示,该基因家族不存在鱼类特有的基因,在硬骨鱼类中具有一定的保守性。比较基因组学结果显示,与四足动物相比,大多数硬骨鱼类中IL17受体基因没有明显增多。鲤与草鱼等其他硬骨鱼类相比,除IL17RB以外,其余IL17受体基因家族成员均加倍。不同组织的表达分析结果显示全基因组复制后不同基因拷贝的功能发生了分化。研究表明,虽然硬骨鱼经历第三轮全基因组复制,但是由于复制发生时间久远,大多数基因已经发生改变或退化,进而在基因组中丢失。而鲤第四轮基因组复制时间发生在820万年前,复制发生时间较近,故复制后的基因基本得以保留。但是对于一些具有特殊功能的高度保守基因(例如IL17RB),也会发生在极短时间内出现丢失现象。鲤和草鱼健康组织的表达谱分析结果同样表明,鲤IL17受体基因的不同拷贝之间已经发生了快速进化及亚功能化,并且这种现象在鲤四倍体基因组中普遍存在。     

大黄鱼肿大细胞病毒不同毒株的细胞培养及主要衣壳蛋白基因比较

为建立大黄鱼肿大细胞病毒的培养方法,明确其分类地位,用肿大细胞病毒检测呈阳性的大黄鱼幼鱼病料 (FD201807和SA201808)肾组织匀浆液感染鳜仔鱼细胞系 (mandarin fish fry cell line-1,MFF-1)并连续传代,从病料组织匀浆液和细胞冻融液中提取病毒DNA,克隆病毒主要衣壳蛋白基因 (mcp),测序后与NCBI GenBank中的虹彩病毒科肿大细胞病毒属病毒mcp以及2018—2020年所检出的15株大黄鱼肿大细胞病毒mcp进行比对分析。结果显示,病毒传至第4代才可引起MFF-1细胞病变,细胞病变的主要特征为细胞脱壁、变圆、折光度增强;感染时间越长脱壁细胞越多,同时培养液中的颗粒增加;透射电镜下可见感染细胞的细胞质散在大小为130~150 nm的六边形病毒粒子和空壳。感染细胞的病变周期随传代代次的增加而缩短,第15代次的FD201807株感染细胞80%细胞病变的时间为3 d,第15代次的SA201808株感染细胞80%细胞病变的时间为7~8 d。mcp序列比对和聚类分析发现,SA201808株与FD201807株的mcp序列存在21个碱基差异,二者的mcp序列分别与大黄鱼虹彩病毒(large yellow croaker iridovirus, LYCIV) LYCIV-Zhoushan (GenBank: MW139932.1)和花鲈虹彩病毒 (Lateolabrax maculatus iridovirus, LMIV) (GenBank: MH577517.1)相近。15株从大黄鱼病料检出的肿大细胞病毒中,12株的mcp序列与SA201808株聚类;3株与FD201807聚类。本研究利用MFF-1细胞系分离培养了大黄鱼肿大细胞病毒,揭示了大黄鱼肿大细胞病毒存在差异,为更好地了解大黄鱼肿大细胞病毒提供了数据参考。     

条石鲷鳃的组织发育及鳃上钠钾三磷酸腺苷酶活性的早期变化

为完善条石鲷的早期组织发育过程,探索条石鲷仔鱼早期高死亡率的内在发育机制,实验对鳃器官的早期发育进行观察,结果显示:条石鲷仔鱼在1日龄时出现鳃原基;2日龄,原始鳃弓形成;3日龄,鳃丝原基出现;6日龄,鳃丝两侧的扁平细胞向外突出而形成原始的鳃小叶,假鳃出现;7日龄,鳃小叶基部出现泌氯细胞;8日龄,泌氯细胞开始分泌蛋白样物质;14日龄,鳃的结构分化发育完善.钠钾三磷酸腺苷酶(Na+,K+-ATPase)的相对活性从胚胎分裂期不断增加,直至器官分化期.此后活性逐渐降低,并保持在一个较低的水平,直至9日龄活性才开始上升.而12日龄之后又不断下降,到14日龄降至最低值(0.246±0.126) U/g.此后急剧上升,到18日龄升至最高值(4.731±0.309) U/g.从20日龄起,Na+,K+-ATPase的相对活性保持稳定上升,直至本次实验结束.结果表明,Na+,K+-ATPase活性的变化与鳃组织,尤其是与泌氯细胞的发育密切相关,而且在其活性较低的阶段,条石鲷仔鱼的死亡率较高.     

Domestication affects spawning of the ide (Leuciscus idus)—preliminary study

Ide, Leuciscus idus, breeders in Polish fish farm are cultured or are collected from the wild. The data about differences in reproduction of different ide forms in captivity are limited. In the present paper domesticated and wild stock are reproduced artificially. Cultured fish do not require application of spawning agents. For obtaining good-quality oocytes only stimulation with temperature and photoperiod is necessary.     

珠江三角洲地区大口黑鲈池塘养殖系统的能值评估

为评价珠江三角洲地区大口黑鲈池塘养殖系统的生态经济性能,实验以能值理论为基础,定量分析该系统的能流和物流特点,通过建立能值评价指标体系,综合评估该系统的环境影响及可持续性。结果显示,大口黑鲈养殖系统投入的资源分为可更新资源(太阳能、风能、雨水能、地球循环能和河水能)和购买的外部资源(设施、苗种、电能、饵料、药品、劳动力、租金、维护费)两部分。养殖系统投入的总能值为4.51×10~(17)sej/(hm~2·a),其中可更新资源能值总和为1.24×10~(16) sej/(hm~2·a),占总投入能值的2.75%。河水能在可更新资源中所占比例最大,为9.77×10~(15) sej/(hm~2·a),占总投入能值的2.17%。购买的外部资源能值总和为4.38×10~(17) sej/(hm~2·a),占总投入能值的97.25%。饵料投入在购买的外部资源中所占比例最大,其能值为3.49×10~(17) sej/(hm~2·a),占总投入能值的77.33%,其次是劳动力,能值为2.29×10~(16) sej/(hm~2·a),占总投入能值的5.08%。大口黑鲈池塘养殖系统太阳能值转换率TR为2.18×106 sej/J,产出能值交换率EERY为2.028,能值产出率EYR为1.028,环境负载率ELR为35.39,能值持续性指数ESI为0.029,可持续性发展的能值指数EISD为0.059。大口黑鲈池塘养殖系统经济效益较高,但过多依赖购买的外部资源,对环境压力较大,可持续性较差。减少饵料投喂量、提高饵料利用率(如选择优质配合饲料及添加剂、改进投喂策略等)以及开展综合养殖是提高珠江三角洲地区大口黑鲈养殖系统持续性、减小环境负载率的有效途径。     

锌对团头鲂幼鱼血清1H NMR代谢组的影响

为了发掘团头鲂对锌(Zn)敏感的代谢标志物,以3种Zn含量分别为7.4、32.1和332.4 mg/kg的半纯化饲料(分别标记为L、M和H组),喂养初始体质量为(3.6±0.1)g团头鲂12周,应用1H(氢谱)核磁共振波谱法(1H NMR)检测团头鲂血清代谢物的种类和浓度,再用最小二乘法判别(PLS-DA)和变量重要性(VIP)分析3个处理团头鲂血清的代谢轮廓和主要差异代谢物。结果显示,血清中52种代谢物被定性和定量,其中氨基酸及其衍生物23种、有机酸16种、糖类4种、核酸组分4种、维生素3种和其他组分2种,经PLS-DA和VIP分析,3组的代谢轮廓相异,其中L组血清中脯氨酸、丙氨酸、赖氨酸等8种氨基酸和葡萄糖含量较M组高,H组血清中则有12种氨基酸和葡萄糖含量较M组高。差异代谢物依次为脯氨酸、乳酸、葡萄糖、丙氨酸、赖氨酸、谷氨酰胺、肌酸、反式-4-羟基-L-脯氨酸、牛磺酸、丝氨酸、τ-甲基组氨酸、精氨酸、缬氨酸、蛋氨酸和甘露糖。以上结果显示,饲料中Zn缺乏和过量会抑制团头鲂的氨基酸代谢和糖代谢,脯氨酸、乳酸和葡萄糖是团头鲂对Zn敏感的潜在生物标志物。