Intraperitoneal injection urocortin-3 reduces the food intake of Siberian sturgeon (Acipenser baerii)

Urocortin-3 (UCN3), one of the corticotropin releasing factor (CRF) family peptides, which was discovered in 2001, has a variety of biological functions. However, the researches of UCN3 in fish were scarce. In order to understand whether UCN3 play a role in regulating food intake in fish, we first cloned the ucn3 cDNAs sequence of Siberian sturgeon (Acipenser baerii Brandt), and investigated the ucn3 mRNA levels in 11 tissues. The Siberian sturgeon ucn3 cDNA sequence was 1044 bp, including an open reading frame (ORF) of 447 bp that encoded 148 amino acids with a mature peptide of 40 amino acids, a 5ʹ-terminal untranslated region (5ʹ-UTR) of 162 bp and a 3ʹ-terminal untranslated region (3ʹ-UTR) of 435 bp. The result of tissue distribution showed that ucn3 widely distributed in 11 tissues with highest expression in brain. We also assessed the effects of periprandial (pre- and post-feeding), fasting and re-feeding on ucn3 mRNAs abundance in brain. The results showed the expression of ucn3 mRNA in brain was significantly elevated after feeding, decreased after fasting 17 days and increased after re-feeding. To further investigate the food intake role of UCN3 in Siberian sturgeon, we performed intraperitoneal (i.p.) injection of Siberian sturgeon UCN3 (SsUCN3) with three doses (60, 120 or 240 ng/g) and recorded the food intake. Acute and chronic i.p. injection SsUCN3 reduced the food intake in a dose-dependent pattern. In conclusion, this study indicates that SsUCN3 acts as a satiety factor to inhibit the food intake of Siberian sturgeon.     

The evidence of apelin has the bidirectional effects on feeding regulation in Siberian sturgeon (Acipenser baerii)

Apelin is a peptide, mainly produced in the brain, which participates in several physiologic effects. However, knowledge about the mechanism of appetite regulation in teleosts, including the role of apelin is not well understood. The aim of this study is to explore the effect of feeding status on the expression of apelin mRNA in the whole brain and the effects of injection of apelin on food intake in Siberian sturgeon (Acipenser baerii). In this study, we first cloned the apelin cDNA sequence of the Siberian sturgeon. We obtained a 1046-bp cDNA fragment, including a 237-bp open reading frame (ORF) that encoded 78 amino acids. Apelin was widely distributed in 11 tissues related to feeding regulation, with the highest expression in thewhole brain, followed by the spleen and trunk kidney. In addition, we measured the effects of periprandial (preprandial and postprandial) change, fasting and re-feeding on apelin mRNA expression in whole brain. The level of apelin mRNA was significantly decreased 1 h after feeding. The results of the fasting experiment showed that the expression of apelin mRNA in the brain was significantly reduced after 1 day of fasting but consistently increased throughout the 15-day food deprivation period. When the 15-day fasted fish were re-fed, apelin mRNA expression in the brain was significantly increased as compared to that of the control. These results suggest that apelin may play a bidirectional role in the regulation of food intake in the Siberian sturgeon. In order to further examine the effect of apelin on feeding regulation in Siberian sturgeons, acute and chronic intraperitoneal (i.p.) injection experiments were performed and food intakes were recorded. Results showed that acute i.p. injection of apelin-13 reduced food intake, however, chronic i.p. injection apelin-13 increased the food intake for 7 days in Siberian sturgeons. In conclusion, our results show that apelin has a bidirectional effect on feeding regulation in Siberian sturgeons by acting as a satiety factor in short-term feeding regulation and a starvation factor in long-term feeding regulation.     

西伯利亚鲟致病性嗜水气单胞菌外膜蛋白及其抗原性分析

采用十二烷基肌氨酸钠(Sarkosyl)法提取西伯利亚鲟嗜水气单胞菌(Aeromonas hydrophila)外膜蛋白,电泳显示所提取的主要外膜蛋白分子量为26～120 kDa;为比较该菌株与气单胞菌菌属其他细菌外膜蛋白组分及抗原性异同,以致病性豚鼠气单胞菌(A.caviae)、温和气单胞菌(A.sobria)和无致病力的嗜水气单胞菌为对照,电泳图谱显示4种气单胞菌外膜蛋白的分子量主要集中在26～120 kDa之间;利用抗西伯利亚鲟嗜水气单胞菌血清的免疫印迹试验表明该菌株外膜蛋白中分子量为75 kDa、52 kDa、43 kDa、40 kDa、34 kDa、28 kDa的蛋白条带呈现阳性反应,其他3种气单胞菌外膜蛋白中均有与该抗血清反应的条带,且分子量为28 kDa、34 kDa的反应条带为4株菌共有;43 kDa与75 kDa反应条带为部分菌株共有.为进一步筛选和研究致病性气单胞菌的共同保护抗原提供参考.     

Morphology and ultrastructure of Siberian sturgeon (Acipenser baerii) spermatozoa using scanning and transmission electron microscopy

BACKGROUND INFORMATION: Available data concerning the sperm morphology of teleost fishes demonstrate wide variation. In the present study, the spermatozoa of Siberian sturgeon (Acipenser baerii Brandt, 1869), a chondrostean fish, was investigated. In contrast with teleost fish, chondrostean spermatozoa have a head with a distinct acrosome, whereas other structures, such as a midpiece and a single flagellum, are present in spermatozoa of most species. RESULTS: The average length of the head including the acrosome and the midpiece was 7.01+/-0.83 microm. Ten posterolateral projections derived from the acrosome were present on a subacrosomal region, with mean lengths of 0.94+/-0.15 microm and widths of 0.93+/-0.11 microm. The nucleus consisted of electrodense homogeneous nuclear chromatin. Three intertwining endonuclear canals, bound by membranes, traversed the nucleus longitudinally from the acrosomal end to the basal nuclear fossa region. There were between three and six mitochondria, two types of centrioles (proximal and distal) in the midpiece and two vacuoles composed of lipid droplets. The flagellum (44.75+/-4.93 microm in length), originating from the centriolar apparatus, had a typical 9+2 eukaryotic flagellar organization. In addition, there was an extracellular cytoplasm canal between the cytoplasmic sheath and the flagellum. CONCLUSIONS: A principal components analysis explained the individual morphological variation fairly well. Of the total accumulated variance, 41.45% was accounted for by parameters related to the head and midpiece of the sperm and the length of the flagellum. Comparing the present study with previous studies of morphology of sturgeon spermatozoa, there were large inter- or intra-specific differences that could be valuable taxonomically.     

Advances in the larval rearing of Siberian sturgeon

Since first large-scale attempts to culture sturgeon from the larval stage were carried out in the former U.S.S.R. at the end of the nineteenth century, rearing technology has advanced considerably during the last 20 years and noticeable improvements in incubation and larval rearing techniques have been implemented in normal hatchery procedures. Siberian sturgeon eggs are incubated in MacDonald jars at 13–14° C to prevent fungal infections. Mass hatching takes place 7 days after fertilization and hatching rate can be predicted as a function of the percentage of eggs fertilized. Survival at the end of the endogenous feeding stage is correlated with hatching rate. Egg size has no direct implications for larval growth and survival of Siberian sturgeon. Experimental studies have demonstrated that behavioural observations are useful criteria to assess the quality of larvae and to synchronize the physiological state of fish with the appropriate rearing procedures. Special attention should be given to the transition to exogenous feeding, where cannibalism, difficulties in adaptation to a new diet, overfeeding and resulting bacterial infections dramatically reduces survival to the fingerling stage. Although a commercial artificial diet specifically formulated for larvae of Siberian sturgeon and other acipenserids is still lacking, commercial non-purified rainbow trout diets and starter marine fish diets are currently used and their results are reasonably acceptable in terms of larval growth and survival. Further research must be focused on the determination of egg quality indicators in order to provide the producer with the tools to estimate the viability and performance of theprogeny.     

How different rearing temperatures affect growth and stress status of Siberian sturgeon Acipenser baerii larvae

Environmental temperature is one of the critical factors affecting fish development. The aim of this study was to examine the impact of three different rearing temperatures (16, 19 and 22°C) throughout the endogenous feeding phase of the Siberian sturgeon Acipenser baerii. This was performed by assessing (a) larval survival and growth; (b) immunofluorescence localization and expression of genes involved in muscle development and growth – myog and Igf1; and (c) stress status through the expression of thermal stress genes – Hsp70, Hsp90α and Hsp90β – and whole body cortisol. Overall survival rate and larval weight did not differ significantly across temperatures. Larvae subjected to 22°C showed faster absorption of the yolk-sac than larvae subjected to 19 or 16°C. Both at schooling and at the end of the trial, larvae reared at 16°C showed significantly lower levels of cortisol than those reared at 19 or 22°C. IGF-1 immunopositivity was particularly evident in red muscle at schooling stage in all temperatures. The expression of all Hsps as well as the myog and Igf1 genes was statistically higher in larvae reared at 16°C but limited to the schooling stage. Cortisol levels were higher in larvae at 22°C, probably because of the higher metabolism demand rather than a stress response. The observed apparent incongruity between Hsps gene expression and cortisol levels could be due to the lack of a mature system. Further studies are necessary, especially regarding the exogenous feeding phase, in order to better understand if this species is actually sensitive to thermal stress.     

Characterization,tissue distribution of resistin gene and the effect of fasting and refeeding on resistin mRNA expression in Siberian sturgeon (Acipenser baerii)

Resistin as an adipokine identified from rodents in 2001 is involved in many biological processes. However, little is known about this gene in fish. We cloned Siberian sturgeon (Acipenser baerii) resistin cDNA of 795 base pairs, encoding 107 amino acids, which showed 38–40% identity to human and rodents. Real-time quantitative PCR showed that the resistin gene was widely distributed in tissues of Siberian sturgeon, with the highest expression in liver. After fasting for 1, 3, 6 and 10 days, the expression of the resistin gene in the liver of Siberian sturgeon decreased significantly, and after refeeding on the 10 days of fasting the resistin mRNA expression increased rapidly, suggesting that resistin may play an important role in liver in response to starvation. Taken together, these results suggest that resistin may be involved in the regulation of energy homeostasis in liver.     

西伯利亚鲟(Acipenser baerii)致病性维氏气单胞菌的分离鉴定

摘要：【目的】本研究旨在寻找引起养殖西伯利亚鲟鱼(Acipenser baerii)病害的致病因子。【方法】从北京地区自然患病的西伯利亚鲟鱼体内分离到致病菌株X-1-06909,采用生理生化鉴定结合16S rRNA基因序列的系统发育学分析确定该菌株的系统发育地位。同时采用琼脂扩散法对抗菌类药物的敏感性进行测定。【结果】菌株X-1-06909与Aeromonas veronii ATCC 35624T的16S rRNA基因序列相似性达99.6％;结合形态特征与生理生化测定结果,革兰氏阴性杆菌,具极生单鞭毛     

Variations of Some Physiological and Immunological Parameters in Siberian Sturgeon (Acipenser baerii,Brandt, 1869) Subjected to an Acute Stressor

This study was carried out to investigate the effect of an acute stressor on the variation of some physiological and immunological parameters of Siberian sturgeon (Acipenser baerii) juveniles. Fish, reared in 3 tanks for 10 weeks, were used for this study. The acute stress of fish consisted of 2 min of air exposure stress. Plasma levels of cortisol, glucose, and lactate as well as lysozyme activity in plasma were measured before stress and 1 hr, 3 hr, 6 hr, 9 hr, 12 hr, and 24 hr after stress. The plasma cortisol significantly increased in the highest level 1 hr after stress, yet it gradually declined after 3 hr. The glucose significantly increased only 1 hr after stress. There was no significant difference between plasma lactate prestress and poststress. Moreover, lysozyme activity was enhanced by stress, thus reaching the highest level 9 hr after stress. The results of this study indicate that Siberian sturgeon not only have a rapid response to acute stress, but also a great capacity for recovery from stress, thus returning physiological parameters to prestress levels after 6 hr.     

Derivation of the clonal‐cell lines from Siberian sturgeon (Acipenser baerii) head‐kidney cell lines and its applicability to foreign gene expression and virus culture

This study was conducted to establish and characterize the clonal‐cell lines from Siberian sturgeon Acipenser baerii head‐kidney tissues and to evaluate its applicability as a research tool. From the culture of A. baerii head‐kidney derived cells, 10 cell lines were established first and then eight clonal‐cell lines were derived from clonal growth and colony expansion of two cell lines that showed significant high colony‐forming ability. All eight clonal‐cell lines were morphologically similar and grew stably under monolayer culture but their growth rates were significantly different. They possessed diploid DNA contents, expressed epithelial cell‐related genes and showed strong anchorage dependency to substrates. When a clonal‐cell line was transfected separately with three plasmid vectors including fluorescent reporter genes driven by cytomegalovirus, marine medaka Oryzias dancena β‐actin or A. baerii β‐actin promoter, the cell lines expressed fluorescent signals regardless of promoter types. The cells harbouring foreign genes could be expanded to stable cell lines under drug selection and then they additionally could form the extensively proliferating colonies at low‐density culture. Finally, the clonal‐cell lines showed the susceptibility to viral haemorrhagic septicaemia virus (VHSV). Collectively, the clonal‐cell lines from A. baerii head kidney were established and these cell lines will be able to provide an excellent in vitro system for various biological studies in this fish species.     

The leaf extract of Siberian Crabapple (Malus baccata (Linn.) Borkh) contains potential fatty acid synthase inhibitors

The present work focused on the kinetics of the inhibitory effects of the leaf extract of Siberian Crabapple, named Shan jingzi in China, on chicken liver fatty acid synthase. The results showed that this extract had much stronger inhibitory ability on fatty acid synthase than that from green teas described in many previous reports. The inhibitory ability of this extract is closely related to the extracting solvent, and the time of extraction was also an important influencing factor. The inhibitory types of this extract on diffeerent substrates of chicken liver fatty acid synthase, acetyl-CoA, malonyl-CoA and NADPH, were found to be noncompetitive, uncompetitive and mixed, respectively. The studies here shed a new light on the exploration for inhibitors of fatty acid synthase.     

不同开口饵料对西伯利亚鲟仔鱼生长、存活和体成分的影响

分别采用水丝蚓（Limnodrilus sp.）、卤虫无节幼体（Artemia nauplii）、枝角类（Moina sp.）和人工饲料饲养西伯利亚鲟仔鱼30 d,研究不同开口饵料对西伯利亚鲟仔鱼生长、存活率和体成分的影响.结果表明：卤虫无节幼体为西伯利亚鲟最适开口饵料,仔鱼的存活率最高（96.67%）; 投喂水丝蚓组生长速度最快;而人工饲料组生长速度明显低于其他组,且成活率最低.不同开口饵料组间仔鱼体成分差异显著,人工饲料组水分含量最高,且粗蛋白和粗灰分含量最低.采用卤虫无节幼体为西伯利亚鲟仔鱼开口饵料,然后采用水丝蚓进行强化培育,能获得较好的生长速度和存活率.     

Oil content,fatty acid composition and biodiesel properties among natural provenances of Siberian apricot (Prunus sibirica L.) from China

Siberian apricot (Prunus sibirica L.), an excellent woody oil plant unique to Asia, is well known for its ability to produce high‐oil seeds for use as a promising feedstock of biodiesel. Based on the investigation of natural Siberian apricot resources in China in the early stage, seeds of Siberian apricot from 74 geographic provenances which can fully reflect the overall information were collected. In this research, seeds oil content, fatty acid composition and biodiesel properties were evaluated, and the key environmental factors that caused the variation of these in different geographic provenance were analyzed. The oil content of Siberian apricot seeds is 45.48%–61.07%, and the average was 50.95% for all provenances. The characteristics of oil can identify and quantify eight fatty acids. The most abundant fatty acids were oleic acid (C18:1; 54.02%–76.54%), followed by linoleic acid (C18:2; 16.78%–38.49%) and erucic acid (C16:0; from 3.27% to 6.12%). Monounsaturated fatty acids are the most abundant in 54.75%–77.03% compared with saturated fatty acids and polyunsaturated fatty acids. The biodiesel properties of most provenance seeds meet the standards of the ASTM D6751 and GB/T 20828, and a few meet the standards of the EN14214. Through the clustering of oil content and fatty acid composition and the analysis of biodiesel properties indexes, it is concluded that KSK provenance is the most suitable for biodiesel production. The XBZ, HHE, AES, ZLQ and LD provenances may be preserved as potential biodiesel. RDA and VPA showed that the effects of environmental factors on the oil properties of Siberian apricot were ranked as terrain factor > climate factor > soil factor, among which longitude, latitude and altitude are the main terrain indicators. These evaluations can provide reference for the effective utilization and further development of Siberian apricot as a bioenergy feedstock.     

Effects of stocking density during live transportation on haematological parameters of Siberian sturgeon (Acipenser baerii,Brandt, 1869)

In this study, we investigated the effects of different stocking densities water quality and blood parameters during the transportation of the Siberian sturgeon (Acipenser baerii). Experiments were carried out in high-density polyethylene tank with three different stocking densities (50, 100 and 150 kg/m³) for 20 h. Ammonium nitrogen (NH₄-N) and nitrite nitrogen (NO₂-N) were measured in the water samples. The erythrocyte, leucocyte, haematocrit (HCT), haemoglobin (Hb), cortisol, and sodium ion (Na⁺) were measured in blood samples. Ammonium nitrogen and nitrite nitrogen concentrations in the water have increased in parallel with the stocking density depending on the time. In the highest stocking density, the maximum levels of NH₄-N and NO₂-N at the 16^th h reached 2.64 mg/L and 4.74 mg/L, respectively. Erythrocyte, leucocyte, HCT, and Hb values did not differ significantly between the experimental groups (p > .05). The results showed that fish could be transported safely for 20 h in 15℃ water temperature at 50 kg/m³ stocking density; however, stocking density of 100 kg/m³ and over could threaten fish welfare and health as from 16^th h.     

Effect of growth temperature and media composition on the fatty acid composition of Bacillus stearothermophilus AN 002

The influence of growth temperature, media composition and cell age on the chemical composition of Bacillus stearothermophilus strain AN 002 has been determined. The total cellular protein decreased and the free amino acid content increased with growth temperature, in both exponential and stationary growth phase. The protein and free amino acid contents of cells were higher in the stationary phase than in the exponential phase, irrespective of growth temperature and media composition. The RNA content was only reduced in cells grown at 55° C. No significant variations were observed in the DNA and carbohydrate contents with respect to growth temperature and cell age. The total lipid and fatty acid compositions on the other hand varied as a function of growth temperature, cell age and media composition. Differences in the relative concentrations of even, odd and branched chain fatty acids were noticed. Novariation was observed in the antiiso and unsaturated fatty acids with respect to growth temperature. The unique variations in the fatty acid composition and total lipids at the growth temperature of 50° C and their variations in the stationary growth phase seem to be characteristic for B. stearothermophilus AN 002.     

Verification of meiotic gynogenesis in Siberian sturgeon (Acipenser baeri Brandt) using microsatellite DNA and cytogenetical markers

Diploid gynogenesis was induced in Siberian sturgeon Acipenser baeri using ultra violet (UV)-irradiated bester ( Huso huso × Acipenser ruthenus ) sperm. The higher survival rate of meiotic diploids was noted after 1350 ergs mm⁻² UV irradiation. A total of 80 meiotic diploids of known parentage from two different experimental treatments were screened using microsatellite DNA and cytogenetical analysis, and uniparental transmission in meiotic diploids was confirmed.     

Withdrawn food rate for larvae of Siberian moth on conifers of Siberia

Experimental evaluation of the mass of food withdrawn by larvae of the Siberian moth during their development on larch, cedar pine, fir, spruce, and common pine was carried out. The obtained dependences between the masses of larvae and food withdrawn by them can be directly used to determine the withdrawn food rate of Siberian moth on each of the Siberian conifer species.     

Growth performance,antioxidant and immune status of Siberian sturgeon (Acipenser baeri) fed diets containing green tea extract and oxidized fish oil

The present study was conducted to determine the effects of dietary oxidized fish oil (OFO) and green tea extract (GTE) on growth performance, antioxidant status and immunological parameters in Siberian sturgeon (Acipenser baeri). Fish weighing about 108 g were randomly assigned to one of nine dietary treatments (control diet without OFO and GTE; diets containing only 25 or 50 mg/kg of GTE; diets containing 50 mg/kg of OFO combined with 0, 25 or 50 mg/kg of GTE; diets containing 100 mg/kg of OFO combined with 0, 25 or 50 mg/kg of GTE) with three replicates of 15 fish stocked in fiberglass tanks using a completely randomized design. Growth performance indices, immunological parameters and antioxidant status of all treatment groups were determined using appropriate assays and compared. Results showed that GTE could enhance growth performance. Feeding GTE alone or combined with the OFO could decrease the hepatosomatic index (HSI) and viscerosomatic index (VSI). Serum lysozyme activity and antibody level were increased significantly by GTE alone or combined with OFO compared to the oxidized diets (p < 0.05). Conversely, lipid peroxidation products and antioxidant enzyme activity were significantly lower in GTE treatment groups and those that received combination of GTE and OFO compared to the oxidized diets. Low‐density lipoprotein (LDL) level in GTE groups was lower than other treatment groups (p < 0.05) however, in OFO treatment groups the LDL level in fish serum was significantly increased in comparison with the other treatment groups (p < 0.05). GTE and OFO alone or combined together significantly decreased the levels of triglyceride and cholesterol compared to the control group (p < 0.05). In conclusion, immunological and biochemical parameters were improved by inclusion of GTE into Siberian sturgeon rancid diets which can protect fish from the oxidative damage challenged by dietary oil oxidation.     

Ontogenetic behavior of Siberian sturgeon, Acipenser baerii: A synthesis between laboratory tests and field data

We have reviewed field data and studies on the behavior and development of Siberian sturgeon at early-life intervals and related them to different ecologically relevant environmental factors that may play a role in the distribution, recruitment, and survival of young fish. Four behavioral phases (swimming-up, rheotactism, shoaling, and foraging) are observed from hatching to the juvenile phase. Each behavior is associated with an early-life interval and might allow fish to occupy different river habitats, directly influencing their distribution, survival, and recruitment. River current intensity, substrate typology, food resources, and predation pressure seem to be the most important factors affecting the distribution of Siberian sturgeon free embryos and larvae, while juveniles and adult fish disperse and migrate according to food abundance and reproduction. Mechanisms involved in regulating downstream migration during Siberian sturgeon early life stages are different than those observed in anadromous sturgeon species. In all large Siberian rivers, with the exception of the Lake Baikal, the Siberian sturgeon is represented by population continuums, and in many cases the foraging range also includes the spawning areas. Ontogenetic changes in Siberian sturgeon behavior could be interpreted as a species-specific mechanism to maintain the population continuums described in this species without significant mixture of local populations within the river.