西伯利亚鲟桑葚心病理组织学和病因的初步研究

为了对心脏囊肿状病变的西伯利亚鲟进行病理组织学观察和病因初步探讨,使用病理组织学和电镜观察技术,对患病西伯利亚鲟各器官进行组织病理学观察。结果表明,患病西伯利亚鲟心脏表面可观察到米粒大、息肉状、紫褐色的囊状肿物,似桑葚状,心外膜上皮细胞和结缔组织明显增生,形成囊肿。组织病变为心外膜囊肿,呈动态发展过程,主要表现为囊肿上皮由扁平变为立方上皮;囊肿壁逐渐增厚与分化,使囊腔不断增多变大,腔内有大量未分化的圆球形、梭形等不同成熟度的血细胞,形成血管组织,并浸润着大量游离或团状分布细胞,整个病变类似一种肿瘤型的恶性增生,心肌细胞肿胀,断裂,溶解,坏死;肝细胞脂肪变性并浸润炎性细胞;肾间质和脾脏淋巴细胞增多。鳃小片之间观察到大量细菌侵袭上皮细胞;鳃小片上皮细胞严重肿胀增生。电子显微镜观察到心脏、肝脏、脾脏和肾脏中有大量细菌;该病可能与微量元素有关,所观察到的细菌不是直接病因。本实验对西伯利亚鲟桑葚心病的组织病理变化进行了较为系统的研究,为进一步查明病因、临床诊断防治和致病机理等相关研究提供基础资料。     

西伯利亚鲟海豚链球菌和鲟疱疹病毒Ⅱ型混合感染的诊断与病理损伤

2018年8月,四川彭州与邛崃的养殖西伯利亚鲟发生一种以出血为临床特征,高死亡率的传染病,为明确其病因,本研究对发病鲟的肝脏、肾脏进行病原菌分离、鉴定和鲟疱疹病毒Ⅱ型(AciHV-2)的PCR检测。从患病鲟体内分离到2株G+链状球菌,其16S rRNA序列(MN416231、MN416230)与GenBank中海豚链球菌16S rRNA序列同源性达99%,在以16S rRNA序列构建的系统发育树上,分离菌与海豚链球菌聚为一支;同时基于海豚链球菌lctO基因的特异性PCR检测为阳性,鉴定2株分离菌为海豚链球菌。提取患病鲟肝脏、肾脏组织DNA进行AciHV-2特异性PCR检测,扩增出501 bp的特异性条带,在以AciHV-2 polymerase基因序列构建的系统发育树上,检测样本与AciHV-2聚为一支。病理组织学上,患病鲟的多组织器官发生明显损伤,尤其是肝脏、肾脏、鳃、脾脏和肠的损伤较为严重,表现为明显的变性、坏死、出血以及炎症细胞浸润;电镜下,观察到肝脏、肾脏组织内大量直径200～220 nm的疱疹病毒样颗粒与0.7～0.8μm的链球菌入侵细胞,并导致细胞损伤。综上,诊断患病西伯利...     

超低温冷冻对西伯利亚鲟精子形态结构损伤的观察

为改进西伯利亚鲟精子超低温冷冻保存技术,探讨精子损伤机制,应用扫描和透射电子显微镜,观察了西伯利亚鲟鲜精与冷冻后精子的形态结构.结果显示,经过冷冻保存后精子的形态发生了很大变化.精子顶体长、精子头中部宽、头中部宽与前部宽比值及中段宽与鲜精相比显著增加(P<0.05);中段长度、后外侧延伸物长度比鲜精显著变短(P<0.05).精子经过冷冻后有30.5%的精子在形态、结构上受到不同程度损伤,受损精子显微结构表现为顶体后外侧延伸物与核糅合,顶体内容物丢失;核膜囊泡化、核膜断裂,核内出现空泡;线粒体内嵴弥散,线粒体脱落;鞭毛外膜松弛与鞭毛脱离等.部分受损伤精子出现顶体丢失,中段脱落,鞭毛自中段基部断裂的现象.精子损伤主要集中在膜系统,中心粒等微管系统基本完好.     

史氏鲟、西伯利亚鲟及其杂交种的同工酶分析

采用聚丙烯酰胺凝胶垂直板电泳对史氏鲟、西伯利亚鲟、史氏鲟(♀)×西伯利亚鲟(♂)杂交种、史氏鲟(♂)×西伯利亚鲟(♀)杂交种4个群体5种组织(心、肝、肌、眼、肾)的酯酶(EST)、苹果酸脱氢酶(MDH)和乙醇脱氢酶(ADH)3种同工酶进行了分析.结果表明:3种同工酶在各自群体内具有明显的组织特异性;在4个不同鲟鱼群体间也表现出明显的规律性差异,可作为区分这4种鲟鱼的生化遗传标记.     

西伯利亚鲟白点病的研究

鲟属硬骨鱼纲、辐鳍亚纲、硬鳞总目、鲟形目,是现在存活最原始的一目,其化石始于中生代三叠纪初,在我国主要养殖品种有西伯利亚鲟,俄罗斯鲟和部分杂交鲟。西伯利亚鲟Siberian sturgeon（Acipenser baeri）分布于前苏西伯利亚地区鄂毕河、叶尼塞河、勒拿河、印迪吉尔卡河和科累马河等各大水系,随着养殖规模产量的扩大,     

全人工繁殖西伯利亚鲟的早期胚胎发育

西伯利亚鲟是重要的经济鱼类和进化发育研究材料。对全人工繁殖西伯利亚鲟的早期胚胎发育进行了观察和研究,成熟西伯利亚鲟卵呈球形、黑色、不透明、平均卵径为2.9mm;在水温15.5～18.0℃下,受精卵历时133h开始孵出,所需总积温为2173～2369℃·h;根据对西伯利亚鲟胚胎发育外部形态及典型特征的观察与分析,将其胚胎发育过程划分为合子、卵裂、囊胚、原肠胚、神经胚、器官形成和出膜7个阶段,共34个发育时期,并对胚胎发育各期的主要形态特征进行描述。     

池养西伯利亚鲟的人工繁殖试验

用浅层地下水和河水的混合水将引进的西伯利亚鲟水花鱼苗在水泥池中培育成8龄性成熟亲鱼。在第一次产卵前3～4个月,以2种不同的降温方式使亲鱼越冬。I组17尾雌鱼,17尾雄鱼,水温7～8 ℃;II组19尾雌鱼,20尾雄鱼,水温7～10 ℃。催产前10～15 d将水温升至16～17 ℃。经人工催产,I组10尾雌鱼排卵,14尾雄鱼排精;II组9尾雌鱼排卵,15尾雄鱼排精。雄鱼采用导管法取精,雌鱼切开输卵管手术取卵。试验结果,共获成熟卵19 490 mL,计87.17×10⁴粒,用半干法授精,获受精卵66.22×10⁴粒,受精率17.5 %～90.0 %;孵出鱼苗39.6×10⁴尾,孵化率20.0 %～71.7 %。结果表明,西伯利亚鲟在水泥池中经人工饲养、培育能达到性成熟,可用来进行人工繁殖。     

豆饼部分替换鱼粉对西伯利亚幼鲟生长的影响

豆饼以0、15％、35％、55％比例替换饵料中的鱼粉，添加55％豆饼组同时另加0．5％的蛋氨酸（Met)，配制成5种近似等能等氮试验饵料。饲养平均体重约40g的西伯利亚幼鲟50d。试验结果表明，15％的豆饼添加水平不影响幼鲟生长，其它添加组幼鲟增重率明显低于对照组，饲料系数明显高于对照组。补充蛋氨酸对提高豆饼饵料营养价值有一定的作用。     

氟苯尼考在感染鲁氏耶尔森氏菌的西伯利亚鲟体内的药效学

通过构建鲁氏耶尔森氏菌(Yersinia ruckeri)感染西伯利亚鲟(Acipenser baeri)病理模型,设立阴性对照组(不攻菌不给药)及阳性对照组(攻菌不给药),比较研究了高(50 mg.kg-1)、中(20 mg.kg-1)、低(5 mg.kg-1)氟苯尼考剂量治疗组和氯霉素(10 mg.kg-1)对照组西伯利亚鲟体内的药效学。连续给药3天后,阴性对照组和阳性对照组累计死亡率分别为0%和50%,表明病理模型构建成功;而氟苯尼考低、中、高剂量组和治疗对照组累计死亡率分别为30%、0%、0%、10%。愈后各实验组鱼血液中均无细菌感染;除阴性对照组外,鱼肝、肾组织中均可分离到鲁氏菌,但不致鱼死亡。实验结果表明,氟苯尼考对感染鲁氏耶尔森氏菌的西伯利亚鲟有较好的治疗作用,药效优于氯霉素,可用于治疗鱼类细菌性疾病。     

诺氟沙星在牙鲆体内的残留及消除规律研究

首次报道了诺氟沙星(Norfloxacin，NFLX)在牙鲆体内的残留及消除规律并建立了组织中NFLX的分离提取及检测方法。以30mg／kg剂量连续5d口服灌胃给药，取血液、肌肉、肝脏、肾脏、鳃5种组织，组织中药物先加入pH7．4磷酸盐缓冲液匀浆，再用乙腈振荡提取，反相高效液相色谱法测定其中诺氟沙星的残留浓度，此方法平均回收率71．37％～82．13％，最低检测限达0．005μg/ml。残留研究结果表明，诺氟沙星在5种组织中消除速率快慢不一，肾脏及鳃等非食用组织为诺氟沙星残留的靶组织。若规定可食用组织中的诺氟沙星最大残留限量为50μg／kg，在本实验条件下，建议临床休药期为10d。     

Impact of starvation on digestive enzymes activities and plasma metabolites in Siberian sturgeon (Acipenser baerii,Brandt, 1869)

To increase the current knowledge about the relationship between nutritional status and the digestive capacity of Siberian sturgeon (Acipenser baerii), we addressed the effect of starvation‐refeeding and macronutrient composition on growth parameters and key digestive enzyme activities in A. baerii. Acipenser baerii juveniles were fed four different diets for 3 weeks, then starved for 2 weeks and allowed to refed for 5 weeks with the same diets. Another group of fish were fed 10 weeks with the corresponding diets. Among 10‐week fed fish, high‐protein diets promoted higher body weight values, while the lowest specific growth rate was observed in fish fed a low‐protein, medium‐carbohydrate, high‐lipid diet (p < .05). At the end of the experiment, in fish refed for 5 weeks following a feeding‐starvation cycle and in 10‐week fed animals, the higher levels of blood glucose, triglycerides and cholesterol were found in fish fed low‐protein diets (p < .05). In all treatments, 2 weeks of starvation decreased α‐amylase activity in the intestine (p < .05), while 4 days of refeeding increased lipase (p > .05) and α‐amylase activity in the intestine as well as pepsin in the stomach (p < .05). Our findings suggest that A. baerii maintains a high capacity to digest proteins and lipids after 2 weeks of starvation and that α‐amylase can be used as an indicator of the nutritional status in fish submitted to starvation‐refeeding cycles. Indeed, refeeding with high‐protein and CHO:L ratio diets after starvation could improve the growth rate of A. baerii in culture.     

Osmoregulation in juvenile Siberian sturgeon (Acipenser baerii)

The osmoregulation capabilities of juvenile Siberian sturgeon exposed to three experimental osmolalities (22, 250 and 387 mOsmol kg⁻¹) were studied over a 45-day period. Growth performance, haematological parameters, ion concentrations, gill and spiral valve Na⁺-K⁺-ATPase activities, as well as gill and spiral valve histology, were measured. At the end of the period, the plasma osmolality of fish kept in 250 and 387 mOsmol kg⁻¹ was higher than that of fish kept in 22 mOsmol kg⁻¹. Similar trends were observed in electrolyte concentrations. Spiral valve and gill Na⁺-K⁺-ATPase activity varied with exposure time and environmental salinity. Shortly after being transferred to hyperosmotic media, spiral valve Na⁺-K⁺-ATPase activity fell, while gill Na⁺-K⁺-ATPase activity remained constant. At the end of the experiment, gill Na⁺-K⁺-ATPase activities in fish kept in isosmotic and hyperosmotic media had increased in comparison to those of the control fish. Moreover, spiral valve Na⁺-K⁺-ATPase activities recovered and were similar to those recorded in fish kept in hyposmotic environments. Although some of the morpho-physiological mechanisms were operational in juvenile Siberian sturgeon in their adaptation to hyperosmotic media, fish cannot be considered hyperosmotic regulators as they were unable to maintain their plasma osmolality and electrolyte equilibrium in salinities higher than 250 mOsmol kg⁻¹. This suggests that the culture of juvenile Siberian sturgeon in brackish environments is unlikely to be successful. However, our data indicate that in natural environments, juvenile Siberian sturgeon in migratory populations (Ob and Lena Rivers) would be able to migrate successfully into estuarine brackish grounds with a salinity of up to 9%. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of microsatellite loci in polyploid Siberian sturgeon (Acipenser baeri Brandt) based on uniparental haploids

Haploid gynogenesis was induced in Siberian sturgeon (Acipenser baeri Brandt) using UV‐irradiated bester (hybrid of Huso huso L. and Acipenser ruthenus L.) sperm. The inactivated heterologous sperm was used successfully to produce gynogenetic haploids of Siberian sturgeon. All haploid progeny were screened using microsatellite DNA analysis and uniparental transmission in gynogenetic haploids was confirmed. The objective of this study was to obtain haploids of Siberian sturgeon and describe the inheritance of microsatellite loci in haploid embryos of this species. This initial work, based on inheritance in haploid Siberian sturgeon, suggests that three studied microsatellite loci segregate disomically, three tetrasomically and one octasomically.     

运动功能相关的中华鲟和西伯利亚鲟幼鱼侧视形态比较研究

通过选取11个侧视形态特征度量进行主成分分析(principal component analysis,PCA),同时设置26个标记点进行相对扭曲分析(relative warp analysis,RWA),比较了中华鲟(Acipenser sinensis)和西伯利亚鲟(A.baerii)幼鱼在侧视形态上的差异以及相对扭曲分析和传统多元分析在鲟鱼形态研究中的优劣。结合前期对它们的有氧游泳能力比较结果(西伯利亚鲟比中华鲟强25%),分析了鲟鱼形态的水动力功能。主成分分析比较表明,中华鲟头长、头高、背鳍前基点之后的躯干高度、尾柄长、背鳍前缘长度均显著大于西伯利亚鲟(P<0.05),而尾鳍上叶显著短于西伯利亚鲟(P<0.05)。相对扭曲分析计算样本的几何信息并可视化统计结果,表明中华鲟的吻厚、吻长、头高、头长、躯干后半段高、背鳍前缘长显著大于西伯利亚鲟,而尾柄长和尾鳍上下叶长显著小于西伯利亚鲟(P<0.001)。上述形态特征直接影响着两种鲟鱼的游泳能力。     

Nutrients apparent digestibility coefficients of selected protein sources for juvenile Siberian sturgeon (Acipenser baerii Brandt), compared by two chromic oxide analyses methods

Apparent digestibility coefficients (ADCs) of dry matter (ADC_d), crude protein (ADC_p), energy (ADC_e) and amino acids in selected feedstuffs were determined for juvenile Siberian sturgeon (8.38 ± 0.20 g). The tested feedstuffs were fishmeal (FM), meat and bone meal (MBM), poultry by-product meal, hydrolysed feather meal, fermented feather meal solvent-extracted cottonseed meal and soybean meal. ADCs were determined using a reference diet and test diets at 7 : 3 ratios with 5 g kg⁻¹ chromic oxide (Cr₂O₃) as an inert marker. Fish were reared in a recirculating system and fed to apparent satiation five times daily. Cr₂O₃ in diets and faeces samples were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) and acid-digestion colorimetry (AC) methods, respectively. The results showed that ICP-AES method was more accurate for Cr₂O₃ determination than AC method, and the results determined by ICP-AES method were used in this study. ADC_d and ADC_p of seven tested ingredients were lowest for MBM (59.1 and 84.5%) and highest for FM (79.9 and 94.5%); ADC_e of tested ingredients were from 71.8% for SECM to 93.2% for FM. ADCs of amino acid in test ingredients followed similar trend to the ADC_p. The ADCs of individual amino acids varied from 61.6% (histidine in MBM) to 98.8% (valine in FM).     

Effects of olive pomace on growth performance,digestibility, body composition and fatty acid profile in yearling Siberian sturgeon,Acipenser baerii (Brandt 1896)

Replacement of olive pomace (OP) with wheat flour in diet was studied in diet of yearling Siberian sturgeon (Acipenser baerii). Fish (165 ± 6.8 g) were randomly allocated to 15 fibreglass tanks (15 fish/tank, in triplicate). Fish were assigned to one of five dietary treatments at 19.1 ± 1.5°C for 8 weeks: control diet (crude protein and crude lipid at 514.6 and 188 g/kg_, respectively) and four experimental diets with 20, 50, 75 and 100 g/kg OP to replace wheat flour in the experimental diets. No significant difference was observed in the final weight, feed conversion ratio, specific growth rate, hepatosomatic index, viscera‐somatic index and survival rate among the treatments (p > 0.05). No change was seen in digestibility of protein and lipid in diets containing 20 g/kg and 50 g/kg OP compared to control group, while these values decreased with increasing in OP above 50 g/kg. Digestibility of dry matter and gross energy among the treatments demonstrated no significant difference (p > 0.05). Polyunsaturated fatty acids, especially C22:6n3 (DHA), in the muscle of the fish fed 75 g/kg and 100 g/kg OP in diets were significantly higher than control group (p < 0.05). These data show that OP could be recommended as a substitute for wheat flour in diet of S. sturgeon but its utilization for other species warranted future works.     

Fishmeal can be totally replaced by plant protein blend at two protein levels in diets of juvenile Siberian sturgeon,Acipenser baerii Brandt

An 8‐week growth trial was conducted to evaluate the effects of a plant protein blend (PPB, comprised of soybean meal and wheat gluten meal) as fishmeal (FM) substitute on growth, nitrogen (N) and phosphorus (P) balance in Siberian sturgeon, Acipenser baerii Brandt. Two control diets were formulated to contain two protein levels (400 and 360 g kg^?1). At each protein level, FM was totally replaced by PPB with supplement of crystalline essential amino acids and mono‐calcium phosphate. Growth performance was not affected by dietary protein sources and protein levels. Fish fed the low‐protein diets had higher FI than that of the high protein groups, while fish fed the FM‐based diets had higher FI than that of fish fed the PPB‐based diets. N/P intake of fish showed linear relationship with total N/P loss. The productive P value was significantly affected by dietary protein sources, but no significant effects were shown on productive N value. The results suggest that Siberian sturgeon fed total plant‐based diets with balance of EAA could maintain normal growth performance and reduce total P excretion both at 360 and 400 g kg^?1 protein levels.     

Effects of stocking density on reared Siberian sturgeon (Acipenser baerii) larval growth,muscle development and fatty acids composition in a recirculating aquaculture system

This study evaluated the effects of rearing density on muscle growth and development in Siberian sturgeon (Acipenser baerii) larvae. Three different stocking densities were tested: low (LD, 30 larvae/l), mid (MD, 80 larvae/l) and high (HD, 150 larvae/l) in a recirculating aquaculture system. Larvae were sampled at hatching (T0), schooling (T1) and complete yolk‐sac absorption (T2) stage and were weighed and processed for muscle tissue histometrical analyses and for qualitative morphological study analyses; fatty acid profile was also determined by Gas Chromatography—Flame Ionization Detector analysis. Low‐density larvae presented a higher weight than MD or HD at T2 (p < 0.05). Histometrical analysis revealed that total muscle area was similar at T1 and T2, but higher than T0, while it was lower at HD at schooling (p < 0.05). The fatty acid profile revealed no differences between densities while, during development, there was a selective consumption: sparing or increasing of essential fatty acids to the detriment of their precursors. Our study suggests that lower densities appear to be more suitable to rear Siberian sturgeon in this particular stage of development. Indeed, larvae reared at the lower density were heavier and longer while larvae reared at the higher density showed lower muscle proliferation rate. As a consequence, LD larvae may exert an increase of potential growth at a mid‐long term.     

Biochemical characterization of Siberian sturgeon Acipenser baeri and sterlet Acipenser ruthenus milt plasma and spermatozoa

This paper provides data concerning biochemical composition of milt of two sturgeon species, Siberian sturgeon bred in aquaculture facility of Inland Fisheries Institute in North Poland and sterlet (from two different populations from Danube and Odra). Milt plasma of Siberian sturgeon and sterlet, when compared to teleost fish, is characterized by much lower osmolality (up 70 to 96 mOsm kg⁻¹) and protein concentration (0.24–0.58 g l⁻¹). In spite of the presence of an acrosome and acrosin the antiproteinase activity of seminal plasma was low (12.79–25.40 U l⁻¹). Activities of arylsulfatase and β-N-acetylglucosaminidase were found in spermatozoa. This agrees with the presence of an acrosome in sturgeons sperm. Similarly to mammals, these enzymes are also present in milt plasma. We determined a range of enzymatic activities from the minimal (seminal plasma) to the maximal damage (supernatants obtained after freezing-thawing without cryoprotectant). Activities of arylsulfatase, β-N-acetylglucosaminidase, lactic dehydrogenase and acid phosphatase were released from spermatozoa after freezing-thawing. For this reason they are good potential candidates as a markers of cryoinjury to sperm acrosome and midpiece. This revised version was published online in July 2006 with corrections to the Cover Date.     

Performance,body compositions,input and output of nitrogen and phosphorus in Siberian sturgeon,Acipenser baerii Brandt,as affected by dietary animal protein blend replacing fishmeal and protein levels

An 8‐week growth trial was conducted using a 2 × 3 factorial design to evaluate the effect of substitution of fishmeal (FM) by rendered animal protein blend [APB, comprised of 400 g kg^?1 poultry by‐product meal, 350 g kg^?1 meat and bone meal, 200 g kg^?1 hydrolysed feather meal (HFM) and 50 g kg^?1 spray‐dried blood meal] in diets of Siberian sturgeon, Acipenser baerii Brandt. Two isoenergetic control diets were formulated to contain two different protein levels [high‐protein control (400 g kg^?1), with 483 g kg^?1 of FM] and [low‐protein control (360 g kg^?1), with 400 g kg^?1 of FM]. At each protein level, dietary FM protein was replaced by APB at 75% and 100% levels and supplemented with crystallized essential amino acid under ideal protein concept. The six diets were named as HC, HAPB75, HAPB100, LC, LAPB75 and LAPB100, respectively. No significant differences were found in weight gain rate (WGR) and specific growth rate (SGR), but fish fed with the low‐protein diets showed higher feed intake and feed conversion ratio. Plasma growth hormone and insulin‐like growth factors I of each group were not significantly different (P > 0.05). The whole‐body composition and liver composition were not affected by dietary protein levels, replacement or their interaction. Muscle protein and lipid contents of fish fed with diet LAPB100 were significantly lower than those of HC group. Digestibility of nitrogen (N) and phosphorus (P) were reduced with higher APB inclusion levels, but productive N and P values of all groups were not different. Lower N and P intake induced lower nutrients losses (P < 0.05). The results suggested that dietary protein level could be reduced to 360 g kg^?1 from 400 g kg^?1 without affecting WGR or SGR and significantly reduced nutrients lose. Furthermore, dietary FM protein can be totally replaced by APB in feed formulation either at 400 g kg^?1 or at 360 g kg^?1 protein level.