两种复海鞘油脂中的脂肪酸成分分析

不饱和脂肪酸尤其是ω-3多不饱和脂肪酸是对人类健康具有重要意义的一类营养成分。对海鞘(Aplidium constellatum)和大洋纵列海鞘(Symplegma oceania)两种复海鞘的油脂类成分进行相对含量分析,发现海鞘(A.constellatum)油脂中含有EPA、DHA两种ω-3 PUFAs,含量分别为19.5%和12.7%,大洋纵列海鞘油脂中EPA、DHA的含量分别为18.2%和7.73%。结果表明,两种海鞘油脂均具有较好的开发前景。     

海鞘油脂的提取及其氧化稳定性研究

分别用乙酸乙酯和超临界CO2提取海鞘中的油脂,并对它们的氧化稳定性进行研究。结果表明,超临界CO2提取的海鞘油脂纯度高,EPA和DHA含量均高于溶剂法提取的油脂,而且具有更好的氧化稳定性,40℃氧化诱导期可达13．9h(柄海鞘)和15．9h(玻璃海鞘)。在其中加入抗氧化成分,发现丹参和荔枝草的乙酸乙酯提取物对海鞘油脂具有很好的抗氧化活性,加入0．04％乙酸乙酯提取物,可使海鞘油脂的氧化稳定性提高8～10倍,而且这些抗氧化成分对溶剂法提取的油脂的抗氧化活性更高。     

南极磷虾脂溶性成分的研究进展

南极磷虾脂类营养含量丰富:油脂提取物中二十碳五烯酸(EPA)含量高于15%,二十二碳六烯酸(DHA)含量高于9%,总磷脂含量高于20%,冰鲜的南极磷虾中总虾青素含量高于3 mg/100 g。本文总结了近年来国内外学者对南极磷虾油中的脂肪酸、虾青素、脂溶性维生素、甾醇等脂溶性成分及其功效的研究进展,以及磷虾油提取方法、多种成分稳定性的研究结果,并展望了我国开展南极磷虾脂溶性成分综合利用的前景。     

东海三个海鞘优势种的油脂成分分析

不饱和脂肪酸尤其是ω-3多不饱和脂肪酸是对人类健康具有重要意义的一类成分。海鞘是一类典型的海洋污损生物,对东海3个海鞘优势种皱瘤海鞘、乳突皮海鞘和长纹海鞘的油脂类成分进行相对含量分析比较,发现乳突皮海鞘油脂中含有EPA一种ω-3 PUFA,含量为5.20%;长纹海鞘油脂中含有EPA一种ω-3PUFA和二十碳四烯酸一种ω-6 PUFA,含量分别为12.7%和3.17%;皱瘤海鞘油脂中含有EPA、DHA两种ω-3 PUFAs,含量分别为10.9%和5.74%,以及亚油酸和二十碳四烯酸两种ω-6 PUFAs,含量分别为11.6%和5.94%。与磷虾油文献数据相比较,皱瘤海鞘油脂总体质量与磷虾油相近,显示了ω-3/ω-6 PUFAs较为均衡的特点。表明皱瘤海鞘的多不饱和脂肪酸具有较好的开发利用前景。     

近江牡蛎等3种贝类的脂类成分分析

分析3种贝类的脂类成分,以期为贝类的高值化加工利用提供基础数据。采用氯仿甲醇法提取近江牡蛎、波纹巴菲蛤和马氏珠母贝中的脂质成分,分析脂质中的胆固醇和磷脂含量,采用液液萃取技术分离中性脂和极性脂,并用GC-MS分析其脂肪酸组成。研究结果表明,3种贝类的粗脂肪含量基本在1%左右;脂质成分中胆固醇含量在0.07~0.12 mg/g;脂质中磷脂含量在14%~34%,极性脂含量在27%~45%;脂质中EPA和DHA总含量在12%~22%,近江牡蛎和波纹巴菲蛤脂质中EPA含量高于DHA,而马氏珠母贝脂质中DHA含量高于EPA。     

日本鬼鲉胚胎及卵黄囊仔鱼发育过程中脂肪及脂肪酸特性变化

以野生日本鬼鲉人工催产获得的受精卵为实验材料,定量检测了日本鬼鲉胚胎及卵黄囊仔鱼发育过程中脂肪组成及脂肪酸含量.结果表明:日本鬼鲉胚胎及卵黄囊仔鱼的总脂肪含量为13.85％～ 11.66％,极性脂肪占总脂肪含量为75.39％～ 72.20％.总脂肪及极性脂肪含量在胚胎发育阶段无显著变化,在卵黄囊仔鱼阶段随发育而显著下降.中性脂肪含量在胚胎发育阶段有显著变化,在卵黄囊仔鱼阶段其含量相对稳定.野生日本鬼鲉胚胎及卵黄囊仔鱼总脂肪的主要脂肪酸为DHA(22∶6n-3),16∶0,ARA(20∶4n-6),EPA(20∶5n-3),18∶0和18∶1n-9.总脂肪及极性脂肪的DHA,ARA,EPA含量(mg/gDW)均随胚胎和卵黄囊仔鱼的发育而显著下降,且DHA和ARA含量均在胚胎囊胚期至尾芽期大幅降低.中性脂EPA和DHA含量随发育呈先升后降,其峰值分别出现在初孵仔鱼和2日龄(2DPH)卵黄囊仔鱼.中性脂ARA含量随发育逐步升高,峰值出现在3日龄(3DPH)卵黄囊仔鱼.在胚胎发育前期,总脂肪DHA和ARA相对EPA被选择性消耗,饱和脂肪酸(SFAs)和多不饱和脂肪酸(PUFAs)相对单不饱和脂肪酸(MUFAs)被机体选择性消耗,N-6PUFA相对N-3 PUFA被选择性消耗;在胚胎发育后期及卵黄囊仔鱼阶段,总脂肪EPA相对DHA和ARA被选择性消耗.在胚胎和卵黄囊仔鱼发育过程中,SFAs中16∶0相对18∶0被选择性消耗.胚胎发育后期阶段和卵黄囊仔鱼阶段,总脂肪MUFAs中16∶1相对18∶1被选择性消耗.实验表明日本鬼鲉胚胎及卵黄囊仔鱼发育阶段极性脂肪中DHA、ARA和EPA可以向中性脂肪中转移,胚胎和卵黄囊仔鱼对不同类别脂肪中的重要脂肪酸的消耗具有选择性,且其选择性与发育阶段相关.     

饲料中DHA/EPA值对星斑川鲽幼鱼生长、体组成及血清生理指标的影响

为研究饲料DHA/EPA值对星斑川鲽幼鱼生长、体组成和血液生理指标的影响,实验配制等氮、等能的5种不同DHA/EPA值(0.64、0.97、1.18、1.59和1.91)的饲料,每个比值设3个重复,饲养周期56 d。结果显示:(1)随着饲料DHA/EPA值的升高,星斑川鲽幼鱼增重率、饲料效率、蛋白质效率均呈先上升后下降的趋势(P0.05)。当饲料DHA/EPA值为0.97~1.59时实验鱼增重最快,饲料效率最高。蛋白质效率则在DHA/EPA值为0.97~1.18时达到最高。蛋白质沉积率(protein retention efficiency,PRE)与饲料DHA/EPA值呈显著二次回归关系(y=-1.589 5x2+2.858 3x+45.184;R2=0.910 8,x=饲料DHA/EPA值,y=PRE),当饲料DHA/EPA值大于0.90时呈下降趋势。肝体比呈先下降后小幅回升的趋势(P0.05),在饲料DHA/EPA值为1.18时达到最低,为2.85%,脾脏指数呈显著上升趋势(P0.05),于饲料DHA/EPA值为1.59组最高(0.12%);(2)肝脏粗脂肪含量随饲料DHA/EPA值的增加呈明显下降趋势(P0.05),且在饲料DHA/EPA值为1.18时降到最低,为8.60%,而后又显著上升,但仍显著低于饲料DHA/EPA值为0.64时的水平(13.44%)。二次回归分析(y=5.199 6x2-15.652x+20.866;R2=0.634 8,x=饲料DHA/EPA值,y=肝脏脂肪含量)显示,当饲料中DHA/EPA值为1.51时肝脏脂肪含量最低。脂肪酸分析结果显示,随着饲料DHA/EPA值的升高,肝脏及肌肉中EPA含量均呈线性下降趋势(P0.05),而DHA含量及DHA/EPA均呈直线上升趋势(P0.05)。肝脏和肌肉组织n-3 HUFA总量不受饲料处理的影响(P0.05);(3)血清总蛋白、球蛋白含量在饲料DHA/EPA值为1.59时显著高于其他各组(P0.05),白蛋白在饲料DHA/EPA值为0.64、0.97和1.59水平最高。溶菌酶(LSZ)活性在饲料DHA/EPA值为1.18时达到峰值(P0.05),为2.76μg/mL。谷丙转氨酶(ALT)活性在饲料DHA/EPA值1.91时无显著变化,而当饲料DHA/EPA1.18时,血清谷草转氨酶(AST)活性提高了65%左右。研究表明,在本实验条件下,以增重率为参考指标,采用二次回归(y=-31.066x2+77.26x+76.541;R2=0.957 4,x=饲料DHA/EPA值,y=增重率)分析可得,当饲料脂肪水平为8.3%,n-3 HUFA含量为0.74%时,星斑川鲽幼鱼[初始体质量(31.70±0.12)g]对DHA/EPA值的最适需要量应为1.24。     

东山岛星座短腹海鞘共附生微生物多样性研究

在前期研究发现星座短腹海鞘(Aplidium constellatum)脂溶性提取物具有降血脂功效的基础上,为深入了解该海鞘的生物学背景,开展了其共附生微生物的多样性研究,为进一步从该海鞘及相关共附生微生物中发现可能存在的降血脂活性次生代谢产物奠定基础。通过Illumina Mi Seq高通量测序技术,分析了星座短腹海鞘样品中共附生微生物的物种多样性和物种丰度信息。测序共获得37 911条序列,310个OTU;稀释曲线表明测序深度足够;多样性指数Simpson值为0.373,Shannon值为2.494。样品共附生细菌分属于24个门,优势门有变形菌门(Proteobacteria,78.86%)、放线菌门(Actinobacteria,6.01%)、拟杆菌门(Bacteroidetes,4.67%)、蓝藻门(Cyanobacteria,2.62%)、厚壁菌门(Firmicutes,1.79%)、柔膜菌门(Tenericutes,1.76%)、螺旋菌门(Spirochaetae,1.33%);分属于197个属,优势属有Ruegeria(60.83%)、SAR116_clade科下未知属(3.27%)、Rhodococcus(2.40%)、Cyanobacteria纲下未知属(2.26%)、Candidatus_Hepatoplasma(1.76%)、Pe M15目下未知属(1.61%)、Spirochaetaceae科下未分类属(1.33%)等10个。通过纯培养技术共分离获得15株可培养共附生菌,分属于8个属,分别为弧菌属Vibrio(7株)、希瓦氏菌属Shewanella(2株),以及Pseudomonas、Pseudovibrio、Thalassobius、Exiguobacterium、Flavobacterium、Thalassospira属各一株。本研究首次为星座短腹海鞘提供了准确的共附生微生物背景数据,为进一步开展活性次生代谢产物研究提供了丰富的微生物菌株材料;同时研究还提示该海鞘有水产病原菌感染,在开发利用过程中应格外关注水产病原菌的安全风险防控。     

渤海银鲳成熟卵巢生化成分分析

采用现代生化分析方法,研究了渤海银鲳卵巢组织的生化成分。结果表明:卵巢水分平均含量为68.83%,蛋白质平均含量15.45%,脂肪平均含量12.37%,灰分平均含量1.21%,糖类(碳水化合物)平均含量1.82%;卵巢组织的17种氨基酸中以谷氨酸含量最高,天冬氨酸次之,赖氨酸居于第三位;卵巢脂质中的主要脂肪酸排列顺序如下:C18∶1(n9&7)>C16∶0>C22∶6n3>C16∶1n7>C20∶5n3,饱和脂肪酸(SFA)占脂肪酸总量的18.95%~25.28%,单烯酸(MUFA)占24.67%~31.87%,多不饱和脂肪酸(PUFA)占19.72%~24.26%,EPA和DHA之和占16.27%~20.08%。     

从鳀鱼油中提取高不饱和脂肪酸（EPA和DHA）的研究

分别用尿素附加法和分子蒸馏法从鳀鱼鱼油中提取出高不饱和脂肪酸（EPA和DHA）。用尿素附加法，以相对于原料油1∶1．5、1∶2、1∶2．5倍量的尿素处理而得到的产品中EPA和DHA的含量分别为35．1％、61．0％和70．0％。分子蒸馏法所得到的产品中EPA＋DHA含量达到70．1％，碘价达     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.