Protective effects of propolis on cryopreservation of common carp (Cyprinus carpio) sperm

Cryopreservation of sperm is common procedures in aquaculture, particularly used for routine in artificial insemination. However, these application cause damages and adversely affected sperm motility, viability and consequently lower hatching rates. The objective of this study is to determine whether propolis has an effect on cryopreservation and fertilization ability and to investigate the potential protective effect of propolis on spermatozoa of Cyprinus carpio. Many studies have been done in cryopreservation offish spermatozoa, but none of them contain propolis in extender composition. The extenders were prepared by using modified Kurokura Solution to which 10% Me₂SO added with different levels of propolis (0.2, 0.4, 0.6, 0.8 and 1 mg ml⁻¹) and 10% egg yolk (as a control without propolis). The pooled semen samples diluted at the ratio of 1:9 by the extenders were subjected to cryopreservation. The percentage and duration of motility and fertilization tests of cryopreserved sperm samples have been done immediately after thawing and compared with control and fresh semen. The extenders containing propolis exhibited higher percentage motility and motility duration than control group (P < 0.05). Especially the group IV (0.8 mg ml⁻¹ propolis) and the group V (1 mg ml⁻¹ propolis) showed significant positive effects on both post thaw motility and hatching ability. The propolis maintained the integrity of the spermatozoa during the cryopreservation process. Evaluating with its contents, it has been shown that propolis is an appropriate cryoprotective agent in fish semen.     

元江鲤种群遗传多样性

选择12对微卫星标记检测了于2011年采集自元江(红河上游中国江段)5个样点192尾鲤的群体遗传多样性.共检测到201个等位基因,每个位点等位基因2-27个.各群体各位点平均等位基因(NA)12.25-14.67个,平均有效等位基因(NE)8.28-9.73个,平均观察杂合度(Ho)o.7765-0.8037,平均期望杂合度(HE)0.7761-0.8080,平均多态信息含量(PIC)0.7534-0.7843.元江鲤种群192个个体各位点NA、NE、Ho、HE、PIC分别为16.50、11.26、0.7927、0.8049、0.7966,种群遗传多样性水平高.元江鲤群体之间遗传分化小,可作为一个种群管理单元进行管理.增殖放流要防止遗传多样性丧失.     

小浪底水库排沙对黄河鲤鱼的急性胁迫

为探讨黄河小浪底水库排沙过程中出现"流鱼"现象的原因,在水库排沙开始前和过程中以黄河鲤鱼为对象进行了生理生态观测。结果显示,相对于对照组,排沙组的黄河鲤鱼消化道充塞系数显著下降(P<0.001),消化道内容物含沙质量比率显著上升(P<0.001);肝脏总抗氧化能力显著上升(P=0.036),而脾脏总抗氧化能力显著下降(P=0.017);肝过氧化氢酶CAT活性显著上升(P=0.035);肾脏超氧化物歧化酶SOD活性显著下降(P=0.038);肝脏、脾脏丙二醛MDA含量显著下降(肝脏:P=0.039;脾脏:P=0.027);鳃丝钠钾ATP酶活性显著下降(P=0.001);血清乳酸LD含量显著上升(P=0.019);肝脏乳酸脱氢酶LDH活性显著下降(P=0.001);推测高含沙水流对黄河鲤鱼造成的急性缺氧应激是导致小浪底水库排沙过程中"流鱼"现象的主要原因之一。     

建鲤leptin两种原核表达载体的构建和表达

使用SignalP-4.0软件预测信号肽剪切位点,设计引物扩增建鲤(Cyprinus carpio var.jian)leptin基因(jlLEP-A1,jlLEP-B)不含信号肽的ORF区域。扩增产物分别连接到pET-32a(+)和pGex-4T-1原核表达载体,构建重组质粒,测序验证插入位点的正确性。将重组质粒转化大肠杆菌BL21(DE3),经IPTG诱导表达重组蛋白pET-32a(+)/jlLEP-A1,pET-32a(+)/jlLEP-B和pGex-4T-1/jlLEP-A1,pGex-4T-1/jlLEP-B。结果表明,在37℃和1 mmol/L IPTG的条件下诱导5 h,重组蛋白表达量最大。SDS-PAGE电泳显示重组蛋白主要以可溶蛋白形式存在。这一结果为后续的建鲤leptin基因功能研究奠定了基础。     

温度和饥饿对鲤鱼幼鱼静止代谢率及自发运动的影响

分别在15和25℃条件下对鲤鱼(Cyprinus carpio)幼鱼[体重(10.36±0.22)g,n=88]进行2周的饥饿处理(饥饿组)或持续投喂(对照组),然后对实验鱼的静止代谢率(resting metabolic rate,RMR)进行了测定,同时以活动期的胸鳍﹑尾鳍摆动频率,观察期(10 min)胸鳍﹑尾鳍平均摆动次数,和运动时间比为指标对实验鱼的自发运动进行了观察。研究发现:温度和饥饿均显著影响鲤鱼幼鱼的静止代谢率(P0.05);在15与25℃条件下饥饿分别导致静止代谢率下降了19%和20%;温度由15℃上升至25℃,对照组与饥饿组静止代谢率分别增加了83%和80%。在15与25℃条件下,饥饿对鲤鱼幼鱼的自发运动具有显著不同的影响(P0.05),且胸鳍和尾鳍的结果十分相似;在15℃饥饿组与对照组的胸鳍、尾鳍摆动频率(活动期,以下类同)均无显著差异,而饥饿组运动时间比显著低于对照组,导致饥饿组的胸鳍、尾鳍平均摆动次数(观察期,以下类同)均显著低于对照组(P0.05);而在25℃饥饿组的胸鳍、尾鳍摆动频率均显著高于对照组,运动时间比无显著差异,导致饥饿组的胸鳍、尾鳍平均摆动次数均显著高于对照组(P0.05)。研究结果提示,在15℃下鲤鱼幼鱼通过降低自发运动以降低能量消耗来适应饥饿,而在25℃条件下,鲤鱼幼鱼通过增加自发运动来增大获取食物的机率以应对饥饿,这可能与野外环境中不同温度下食物丰富度及鲤鱼幼鱼的生理状况有关,但静止代谢率并未呈现与自发运动相关联的变化。     

建鲤IGFBP 3 基因多态性对生长的影响

胰岛素样生长因子结合蛋白3(insulin-like growth factor binding protein,IGFBP3)是调节动物生长和代谢的重要基因.本实验查找了建鲤(Cyprinus carpio var.jian)IGFBP3s基因上的SNP位点.使用PCRRFLP检测了其中5个位点(IGFBP3a-I3...     

Thermal adaptability and disease association in common carp (Cyprinus carpio communis) acclimated to different (four) temperatures

The present study was designed to investigate the effect of temperature (20 °C, 24 °C, 28 °C and 32 °C) on the heamato-biochemical and histological alterations of Cyprinus carpio communis. Increase in the temperature showed significant decrease in the serum protein, while a reduced level of blood glucose at high temperature of 32 °C was observed leading to hypoglycemic conditions in the experimental fishes. A significant correlation (P<0.01) was observed between cholesterol (Cho) and triglycerides (TG) for different temperature treatments. Elevated blood urea nitrogen (BUN) at high temperatures was a good indicator of gill osmoregulatory failure. A variation of 86.40% and 38.33%, respectively, was noticed in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) at 32 °C over minimum experimental temperature of 20 °C. The increase in red blood cell (RBC) and Heamoglobin (Hb) concentration is associated with the decrease of mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC), could be the reason for observed poikilo-anisocytosis. Histological studies of different organs of experimental fishes showed accumulation of MMC's (melanomacrophagic centers) and atrophy of the interrenal tissue on exposure to various levels of temperature. These changes were related to severity of thermal stress, being most marked when high temperature was prolonged during acclimatization. Some fishes were found infested by protozoan parasite at elevated temperature of 32 °C. Increased levels of certain biochemical and haemotological parameters studied were strongly correlated with disease in the Cyprinus carpio communis species.     

Genetic characterization of common carp (Cyprinus carpio) populations from Greece using mitochondrial DNA sequences

Wild common carp from two lakes and two rivers in Greece were genetically characterized with sequencing analysis of two mitochondrial DNA segments: cytochrome b (1119 bp) and D-loop (646 bp). A total of 9 variable singleton sites and 7 unique haplotypes were detected. A common haplotype was found in three out of the four populations examined, which seems to be the ancestral one and represents the European origin of common carp from Greece. This haplotype could be also justified by the introductions reported with individuals belonging to the Central European race, into many natural habitats in Greece. Limited genetic variation — in Evros and Aliakmonas populations — could be due to bottleneck effects and small effective population sizes, whereas the different haplotypes found in Lake Volvi could represent different common carp stocks. Values of sequence divergence among Greek haplotypes ranged from 0.0006 to 0.0023. The Neighbour-Joining (NJ) phylogenetic tree constructed based on the combined sequences, reveals that the populations of common carp from Greece belong to the European group of populations — which is highly divergent from the South East-Asia cluster — and to the subspecies Cyprinus carpio carpio.     

Quantitative genetic estimates of growth-related traits in the common carp (Cyprinus carpio L.): A review

The common carp (Cyrpinus carpio L.) is the oldest cultured and the most domesticated fish species, as well as one of the most important freshwater fishes in the world. However, scientific studies on evaluating the growth-related quantitative traits in this fish are limited. Heritability, the most important parameter in selective breeding programs, was extensively studied for the growth-related traits. The values varied widely among the experiments and methods used because of the existence of common environmental, dominance and maternal effects. However, correlations in phenotypic and genetic levels first evaluated several years ago were limited. On the other hand, heterosis was widely reported and easily obtained for growth-related traits in the common carp. Meanwhile, genotype environment interaction and prediction of breeding values have been studied recently, and are very important in conducting selective breeding programs. The developmental quantitative genetics of growth-related traits was first analyzed in the common carp for reasonable selection during ontogeny. It is expected that genetic improvement will be achieved by carrying out direct selective breeding in the common carp.     

Ice-age endurance: the effects of cryopreservation on proteins of sperm of common carp, Cyprinus carpio L

Damage to spermatozoa during cryopreservation is regarded as a major obstacle to the expansion of sperm storage technology. The authors used two-dimensional polyacrylamide gel electrophoresis and matrix-associated laser desorption/ionization time-of-flight mass spectrometry to explore whether the protein profile of common carp (Cyprinus carpio) spermatozoa is affected by cryopreservation. Fourteen protein spots were significantly altered following cryopreservation. Eleven of these were identified: three as specific membrane proteins (N-ethylmaleimide-sensitive fusion protein attachment protein alpha, cofilin 2, and annexin A4) involved in membrane trafficking, organization, and cell movement; six as cytoplasmic enzymes (S-Adenosylhomocysteine hydrolase, Si:dkey-180p18.9 protein, lactate dehydrogenase B, phosphoglycerate kinase 1, transaldolase 1, and esterase D/formylglutathione hydrolase) involved in cell metabolism, oxidoreductase activity, and signal transduction; and two as transferrin variant C and F. Based on these findings, the authors hypothesize that transferrin in cryopreserved sperm may protect spermatozoa against oxidative damage during the freeze-thaw process. Cryopreservation caused changes in spermatozoa protein profiles that may lead to decreased spermatozoa velocity, motility, and fertilization success, and to subsequent ova hatching rate.     

鲤TYK2基因的克隆鉴定及组织表达分析

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Cloning, characterization, and expression analysis of Toll-like receptor-7 cDNA from common carp, Cyprinus carpio L.

The Toll-like receptor 7 (TLR7) is activated by single strand RNA and RNA-like compounds (imidazoquinoline), and it induces interferon production. We identified and described carp TLR7 cDNA and its mRNA expression. The full-length cDNA of carp TLR7 gene is 3427 bp, encoding 1049 amino acids (AB553573). The similarities of carp TLR7 with zebrafish, rainbow trout, fugu, and human TLR7 were 89.6, 83.4, 80.6 and 74.6%, respectively, at the amino acid sequence level. Furthermore, the expression of TLR7 mRNA was investigated in normal tissues of carp by semi-quantitative RT-PCR analysis. Carp TLR7 expression was exhibited in healthy tissues (kidney, brain, spleen, skin, intestine, muscle, liver, gills and heart) and though the expression level in each tissue varied among healthy fish. Carp TLR7 expression was significantly increased in head kidney stimulated with TLR7 agonist, imiquimod, at 8, 24 and 48 h in vitro when compared to expression in the control group. Moreover, carp head kidney leukocytes produced elevated levels of pro-inflammatory and type 1 interferon cytokine mRNA in response to imiquimod stimulation.     

Percoll gradient separation of cryopreserved common carp spermatozoa to obtain a fraction with higher motility, velocity and membrane integrity

We attempted to select a fraction of common carp, Cyprinus carpio spermatozoa that best survived a conventional freeze/thaw procedure, by centrifugation of frozen/thawed sperm through a Percoll gradient (45% and 90%). The proportion of motile spermatozoa (65.81 ± 5.19%), their velocity (77.58 ± 31.07 μm/sec), and membrane integrity (83.66 ± 4.38% intact) were significantly higher in separated sperm than in whole samples (motility 23.36 ± 2.98%, velocity 55.55 ± 19.03 μm/sec, and membrane integrity 57.92 ± 4.65%). Our results demonstrated that Percoll gradient centrifugation shows promise as a technique for selecting high quality cryopreserved fish spermatozoa, which could be useful for cryobiological research. Further studies are needed to evaluate the potentially higher fertilizing ability of the separated spermatozoa.     

The effects of water temperature and ration size on growth and body composition of fry of common carp, Cyprinus carpio

Experiment was conducted with the aim of determining the effect of varying water temperature and ration size on growth and body composition of fry of the common carp, Cyprinus carpio. Common carp fry with an initial body weight (BW) of 0.86 g were fed a diet (34.9% protein, 18.3 KJ/g diet) at four ration sizes 4%, 5%, 6% and 7% of their body weight per day and reared at two water temperatures 28 and 32 °C for 60 days. Fry fed with 6% ration showed the highest mean final body weight at 28 °C. Final body weight was significantly (P<0.05) affected by ration and temperature. Cyprinus carpio fry raised at 28 °C had higher feed efficiency (FE) (44.36%) than the fry reared at 32 °C (40.98%) with 4% ration. Further, feed efficiency decreased with increase in ration levels in both temperatures. Protein efficiency ratio (PER) was higher (1.26) at 28 °C than at 32 °C (1.17). At 6% ration, common carp fry showed highest specific growth rate (SGR) (3.82%/day) at 28 °C as compared with at 32 °C (3.57%/day). A linear increase in protein and lipid contents was evident with increasing ration levels up to 6% body weight at both temperatures 28 and 32 °C. Second-order polynomial regression analysis of weight gain and SGR indicated the breakpoints at ration level 6.04% and 6.08% body weight per day at 28 and 32 °C. Hepatosomatic index (HSI) not affected by temperature and ration size while, viscerosomatic index (VSI) influenced (P<0.05) by ration size and temperature. Based on the above results, it may be concluded that 6% BW/day ration is optimal for growth of Cyprinus carpio fry at both the temperatures 28 and 32 °C.     

应用SRAP标记分析福瑞鲤及其原始亲本的遗传结构

摘要：应用SRAP分子标记对建鲤（Cyprinus carpio var. jian）、黄河鲤（C.c.haematopterus）和福瑞鲤（FFRC Strain Common Carp,C.carpio）进行遗传结构分析。结果显示,筛选出的10个多态性较好的引物组合共扩增出110个位点,其中多态性位点92个,平均多...     

Biomass-dependent effects of common carp on water quality in shallow ponds

We examined the biomass-dependent effects of common carp (Cyprinus carpio) on water quality in 10 ponds at the Eagle Mountain Fish Hatchery, Fort Worth, Texas, USA. Ponds contained 0–465 kg ha⁻¹ of common carp. We measured limnological variables at weekly intervals for four weeks in early summer, after which ponds were drained and the biomass of fish and macrophytes was determined. Common carp biomass was significantly positively correlated with chlorophyll a, total phosphorus, total nitrogen, and Keratella spp. density and negatively correlated to bushy pondweed (Najas guadalupensis) biomass. In addition, we combined our data with data from comparable studies to develop more robust regression models that predict the biomass-dependent effects of common carp on water quality variables across a wide range of systems.     

Influence of proximal stimuli on swimming in the sea hare Aplysia brasiliana

Although the neurobiology and physiology of sea hares are extensively studied, comparatively little is known about their behaviour or ecology. Several species of sea hares swim, but the function of swimming is unclear. In this paper, we tested the hypotheses that swimming in Aplysia brasiliana serves to find food and mates, and to escape predators. Our data strongly support the hypothesis that swimming in A. brasiliana is related to feeding. Sea hares deprived of food overnight swam 12 times longer than ones that had been fed. When sea hares contacted food while swimming they invariably stopped, while those contacting a plastic algal mimic mostly continued to swim. Our experiments provided no evidence to support the hypothesis that swimming in sea hares is related to social behaviour. Sea hares deprived of copulatory mates for 3 days did not swim longer than ones held in copulating groups. Moreover, swimming sea hares never stopped swimming upon encountering a conspecific. Our experiments also supported the hypothesis that swimming in sea hares is related to predation. Sea hares stimulated with a standardised tail pinch and exposed to ink of conspecifics swam four times longer than control individuals, and tail-pinched sea hares that released ink swam five times longer than ones that did not release ink. However, because predators of adult sea hares are mostly lacking and because sea hares often swim spontaneously without predators being present, we conclude that swimming behaviour in A. brasiliana is primarily related to food-finding.     

Septal pore complex morphology in the Agaricomycotina (Basidiomycota) with emphasis on the Cantharellales and Hymenochaetales

The ultrastructure of septa and septum-associated septal pore caps are important taxonomic markers in the Agaricomycotina (Basidiomycota, Fungi). The septal pore caps covering the typical basidiomycetous dolipore septum are divided into three main phenotypically recognized morphotypes: vesicular-tubular (including the vesicular, sacculate, tubular, ampulliform, and globular morphotypes), imperforate, and perforate. Until recently, the septal pore cap-type reflected the higher-order relationships within the Agaricomycotina. However, the new classification of Fungi resulted in many changes including revision of existing and addition of new orders. Therefore, the septal pore cap ultrastructure of more than 325 species as reported in literature was related to this new classification. In addition, the septal pore cap ultrastructures of Rickenella fibula and Cantharellus formosus were examined by transmission electron microscopy. Both fungi have dolipore septa associated with perforate septal pore caps. These results combined with data from the literature show that the septal pore cap-type within orders of the Agaricomycotina is generally monomorphic, except for the Cantharellales and Hymenochaetales.It appears from the fungal phylogeny combined with the septal pore cap ultrastructure that the vesicular-tubular and the imperforate type both may have arisen from endoplasmic reticulum. Thereafter, the imperforate type eventually gave rise to the perforate septal pore cap-type.     

Mesoderm differentiation in explants of carp embryos

An analysis of carp blastoderm development was carried out in culture after isolation from the yolk cell and its yolk syncytial layer (YSL). The blastoderms were separated from the YSL at four different stages of embryogenesis: the blastula, early epiboly, early gastrula and late gastrula stages. Absence of the YSL in explants was checked by scanning electron microscopy. From observations of living embryos and histological examination of tissues which were formed in explants from all stages studied it was observed that they contained notochordal, muscle and neural tissue as signs of dorsal types of differentiation. Only in explants from the early and late gastrula stages were histotypical tissues organized in an embryonic-like body pattern. The data indicate that mesoderm differentiation in fish embryos is independent from the YSL, contrary to normal pattern formation which needs the presence of the YSL before the onset of gastrulation.