诺氟沙星在西伯利亚鲟体内蓄积规律及其对组织的病理损伤

为了确定诺氟沙星在鲟养殖中的科学使用方法,对不同剂量、不同给药次数情况下诺氟沙星在西伯利亚鲟(Acipenser baerii)体内蓄积规律及毒性进行了研究。分别以0、30 mg/kg、50 mg/kg、100 mg/kg的剂量对西伯利亚鲟连续口灌诺氟沙星3~5 d,每天1次,并于停药后24 h及240 h采集实验鱼血、肝、肾及软骨组织,对4种组织中药物蓄积量进行测定,并对肝、肾和软骨组织进行切片观察。实验结果表明:随着给药剂量增加、给药次数增多,实验鱼4种组织中诺氟沙星浓度逐渐升高,浓度由高到低依次为为:肾、软骨、肝、血清,且血清中药物浓度远远低于另外3种组织。相同给药剂量、不同给药次数时血清中药物浓度无显著差异;而肝、肾和软骨中均差异显著,停药240 h后,显著性消失。切片结果表明,50 mg/kg剂量连续给药5 d后,实验鲟肝、肾组织开始结构损伤变化,100 mg/kg剂量连续给药3~5 d后,损伤情况持续加重,停药240 h后,肝组织损伤情况有逐渐恢复趋势,而肾组织未见明显恢复趋势;而100 mg/kg连续给药3~5 d后,软骨组织出现软骨膜内层细胞减少;停药240 h后有恢复趋势。研究结果表明,诺氟沙星在鲟中使用剂量应小于50 mg/kg,连续给药次数低于5次,以避免对鲟肝组织造成损伤,引起肾不可逆损伤及影响软骨细胞发育和再生,对软骨组织造成潜在影响。     

氟离子对西伯利亚鲟胚胎发育的影响

本试验在水温17±1℃的条件下,采用半静态式生物毒性试验方法,探讨了氟离子(F-)对西伯利亚鲟胚胎发育的影响。结果表明,100~600 mg/L氟暴露会导致胚胎孵化延迟,氟暴露组半数孵化时间(MHT)比对照组推迟9~22 h。氟暴露同时导致胚胎死亡率显著增加(P<0.05),出膜仔鱼活力减弱、存活时间缩短、畸形率显著升高(P<0.05),出现卵黄囊畸形、脊椎畸形、眼部充血等畸形症状。试验得出F-致西伯利亚鲟胚胎144 h半致死浓度(LC50)为447.61 mg/L,孵化安全浓度(SC)为4.48 mg/L,导致仔鱼畸形的半数效应浓度(EC50)为536 mg/L。氟对西伯利亚鲟胚胎的安全质量浓度低于我国部分高氟地区地下水、地表水及人为氟污染水域中氟含量,提示天然水体中氟可能会对鱼类胚胎产生负面影响,应引起重视。     

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.     

Induction of gynogenesis in stellate sturgeon (Acipenser stellatus Pallas, 1771) and its verification using microsatellite markers

Normal egg fertilization was carried out in stellate sturgeon Acipenser stellatus sperm by 60‐, 90‐ and 120‐s ultraviolet irradiation, and a cold shock was carried out afterwards. The treatments included haploids, diploid gynogens, triploid and control groups. Fertilization, hatching and survival rate at the 60‐s irradiation time were significantly greater than those in the 90‐s and 120‐s irradiation and control groups. Two pairs of microsatellite markers were used to find gynogenesis in comparison with the parental and offspring genome in one locus. The results showed all maternal genomes among gynogen offspring with no paternal genome. The markers also showed the maximum gynogenesis induction in the Gy60 treatment.     

西伯利亚鲟鱼池塘养殖试验

2004年8月-2005年2月,笔者在石家庄井陉县进行了西伯利亚鲟鱼池塘养殖,成活率达90％以上,现将养殖情况总结如下。     

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.     

Occurrence and in vitro biosynthesis of 11-ketotestosterone in Siberian sturgeon,Acipenser baeri Brandt maturing females

High levels of 11-ketotestosterone (11KT) were found (49 to 160 ng ml^–1) in plasma of Siberian sturgeon females during the end of their reproductive cycle. These levels were measured either by specific radioimmunoassay, or both by specific radioimmunoassay and by UV absorption after HPLC (isocratic conditions, 33% methanol, 26% acetonitrile, 41% water). In order to find the origin of 11KT synthesis, ovaries were incubated (30 min and 2h at 20°C) with tritiated 17-hydroxyprogesterone (17OHP) or with tritiated androstenedione (A4). Testosterone (conversion rate from tritiated 17OHP: 4%) and 11-ketotestosterone (conversion rate from tritiated A4: 1.6%) were identified as metabolites of respectively 17OHP and A4 (TLC, HPLC and crystallization). 11-hydroxyandrostenedione (11OHA4) and 11-hydroxytestosterone (11OHT) were suggested to be intermediate metabolites. Besides interrenal and blood cells were incubated respectively with tritiated cortisol and tritiated A4. 11OHA4 was identified in interrenal incubation (yield from tritiated cortisol: 1.2%). 11KT in interrenal (yield from tritiated cortisol: 0.14%), and 11OHA4 and 11KT in blood cells (yield from tritiated A4: 1.6%), were suspected to be synthesized (TLC, HPLC, acetylation). No significant metabolization of tritiated cortisol could be found in liver. The possible contribution of each of these tissues to high 11KT levels found in plasma is discussed.To whom correspondence should be sent     

Histochemical identification of the black-brown pigment granules found in the alimentary canal of Siberian sturgeon (Acipenser baeri) during the lecitotrophic stage

The histochemical characteristics of the black-brown pigment granules accumulated during the lecitotrophic stage in the digestive tract of Siberian sturgeon, Acipenser baeri larvae, were studied in order to identify the nature of such pigments. Pigment granules appeared just after hatching in contact with the endodermal epithelium and in the periphery of the yolk-sac matrix, and increased in number with digestive system differentiation. At 3 days post hatch, the accumulation of pigment granules in the spiral valve formed a pigmented plug, which was ejected with first faeces after first feeding. Pigment granules were insoluble in all solubilising solutions tested (distilled water, H₂O₂, NH₄OH, KMnO₄, absolute methanol and different acid, basic and oxidising solutions). Pigment granules were completely bleached with the oxidising processes of Gomori and Casella (15 min) and with H₂O₂ (20%) during 48 h. Histochemical characteristics of bleached pigmented granules revealed that they were basophilic (Eosine negative), presented a moderate methachromasia (Toluidine Blue positive) and argentaffinity (Masson Fontana method positive), and contained carboxylated and sulphated (strongly and weakly ionised) glycoconjugated residues (Alcian Blue pH 0.5, 1.0, 2.5 positive). Bleached pigment granules also contained reductor groups (Ferric ferricyanide-Fe III positive), while they were negative for Bromophenol Blue, Perl's Prussian Blue, Red Oil O and Sudan Black B, indicating the absence of proteins, iron and lipidic substances respectively. According to such results, the black-brown pigment granules observed in the alimentary canal of Siberian sturgeon larvae during the lecitotrophic stage correspond to melanins.     

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.     

Daily patterns of ammonia nitrogen output of Siberian sturgeon Acipenser baeri (Brandt) of different body weights

The daily pattern of ammonia nitrogen outputs of five weight groups of Siberian sturgeon, Acipenser baeri (Brandt), was studied. Regardless of weight, there was a delay of 3 h between the start of feeding and the increase in ammonia output. For sturgeon reared at 17°C with continuous feeding, the total ammonia output levels (NH₄-N + NH₃-N) decreased from 530 mg kg^?1 per day for fish weighing 40 g, to 239 mg kg^?1 per day for fish weighing 1700 g. The study showed that continuous feeding is suitable in sturgeon farms in terms of ammonia loadings.     

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。     

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.     

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.     

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).     

Ascorbic acid status as affected by dietary treatment in the Siberian sturgeon (Acipenser baeri Brandt): tissue concentration,mobilisation and L-gulonolactone oxidase activity

A study was conducted to evaluate tissue storage and mobilisation of L-ascorbic acid (AA) in the Siberian sturgeon (Acipenser baeri) fed three different experimental diets. The three treatments consisted of a diet devoid of vitamin C (diet A0) and two diets supplemented with equivalent of 300 mg AA kg^–1 in the form of either silicone-coated ascorbic acid (diet SC) or of ascorbyl-2-polyphosphate (diet AP). During the first phase (4 months) of the trial, six batches of 130 Siberian sturgeon (initial body weight: 25.5±0.5 g) each were fed one of the three diets in duplicate. During the second phase (3 months), fish from groups SC and AP were switched to diet A0 and those fed diet A0 during the first phase were switched to diet SC. Irrespective of the dietary treatment, growth rates were not significantly different from each other. At the end of phase I, in all tissues studied, total ascorbic acid (TAA) concentrations were higher in Siberian sturgeon fed diet AP than in the other two groups. During phase II, tissue ascorbate depletion was also higher in the AP group than in the other two groups. Transfer of the AA-free diet fed group onto a diet supplemented with 300 mg AA kg^–1 (diet SC) led to a slight increase in the TAA concentrations in all tissues. Blood plasma tyrosine concentrations were not significantly different between the three groups. Whole-body collagen levels were affected by dietary AA levels or forms at the end of phase I; the differences were not significant at the end of phase II. Muscle collagen levels were slightly affected. L-Gulonolactone oxidase activity was found in the kidney of Siberian sturgeon, but not in the liver. The ascorbyl-2-polyphosphate appears to be either better utilised by Siberian sturgeon, like in many other teleosts, or more stable than the silicone-coated AA during food processing and storage. Presence of L-gulonolactone oxidase activity in Siberian sturgeon kidney combined with the absence of gross scorbutic signs in AA-free diet fed groups expressing very good growth rates suggested no need of dietary AA byA. baeri.     

Spirulina as a nutrient source in diets for growing sturgeon (Acipenser baeri)

The efficiency of diets with the inclusion of Spirulina for Siberian sturgeon weaning has been tested. Three isoproteic and isoenergetic diets were formulated with an increasing level of Indian strain Spirulina (SP 40%, SP 50% and SP 60%); the diets were tested against a control diet without microalgae. The results show that Spirulina inclusion improves growth and that an inclusion level of 50% gave the greatest growth rate, a better favourable feed conversion rate and the highest protein efficiency. The fatty acid composition of fillets showed differences between the experimental and control diets: an increase in the Spirulina level induces increases in palmitic and linoleic acids and a decrease in the myristic acid. The control diet was characterized by high levels of eicosapentaenoic and docosahexaenoic acids. At the end of the experiment, statistical differences appeared in the fatty acid profile of the sturgeon fillet, mainly concerning high content of monounsaturated fatty acid and polyunsaturated fatty acid in the sturgeon fillets. If the problems related to the high production costs are solved, Spirulina could prove a good partial substitute fish meal.     

Immunohistochemical investigation of the pituitary of the sturgeon (Acipenser baeri,Chondrostei)

An immunohistochemical study of the sturgeon (Acipenser baeri) pituitary was undertaken using antisera directed against hormones from various classes of vertebrates, including the only pituitary hormone available from sturgeon, gonadotrophin. A positive reaction was obtained after application of antisera towards the following hormones 1–24 synthetic ACTH (1-24 ACTH), melanophore stimulating hormone (MSH), ovine prolactin (oPRL), ovine growth hormone (oGH), salmon growth hormone (sGH), carp gonadotrophin (cGTH) and its beta subunit (cGTH), sturgeon gonadotrophin (aciGTH), carp thyrotrophin (cTSH) and subunit of the human thyrotrophin (hTSH). The results demonstrate that, in general, the sturgeon pituitary resembles that of teleosts as regards the distribution of the different cell types: ACTH and PRL cells in the rostral pars distalis, GTH, TSH and GH cells in the proximal pars distalis and MSH and PAS-cells in pars intermedia. In addition to the topographical organization of the sturgeon pituitary, this study provides data on the immunological relationships between sturgeon pituitary hormones and those of other vertebrates.     

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.     

Verification of ploidy level in sturgeon larvae

Chromosome preparations and assay of the microsatellite locus Afu‐68 were used to determine ploidy in Siberian sturgeon (Acipenser baeri Brandt) and F₁ hybrids of Siberian sturgeon and Russian sturgeon (Acipenser gueldenstaedti Brandt). The chromosome number and microsatellite locus Afu‐68 were compared and these analyses were used for identification of ‘haploid’, ‘diploid’ and ‘triploid’ progeny of the studied cross of A. baeri× (A. baeri×A. gueldenstaedti).