高度不饱和脂肪酸对水生动物生长、发育和繁殖的影响与机理

正高度不饱和脂肪酸是多不饱和脂肪酸中具有二十个碳原子以上,含3个或3个以上双键的脂肪酸~[1-4],主要有花生四烯酸(C20:4n-6)、二十二碳六烯酸(C22:6n-3)和二十碳五烯酸(C20:5n-3)等~[5-12]。高度不饱和脂肪酸影响机体脂类代谢~[13]、基因表达~[14-15]、细胞膜功能~[16]、机体免疫~[17-18]及血液生理生化特性~[19-20]等,对水生动物的生长、发育和繁     

水产品中辛硫磷、倍硫磷和蝇毒磷的超高效液相色谱-串联质谱法检测

[目的]建立了水产品中辛硫磷、倍硫磷和蝇毒磷3种有机磷农药的检测方法。[方法]样品经酸化乙腈提取,PSA吸附剂分散固相萃取净化,超高效液相色谱-串联质谱在多反应离子监测模式下检测,基质匹配外标法定量。结果加标浓度在10~100μg/kg范围内,线性相关系数(R)大于0.99。3种有机磷农药的平均回收率范围为65.8%~101.0%,RSD范围为11.4%~16.0%。本方法检出限为10μg/kg,定量限为20μg/kg。[结论]该方法可用于水产品中辛硫磷、倍硫磷和蝇毒磷等3种有机磷农药残留的同时测定。     

蛭弧菌在水产养殖中应用研究进展

正蛭弧菌是一类专门寄生于其他细菌并能导致其裂解的革兰氏阴性菌,对多种致病菌具有良好的裂解效果~[1],属于蛭弧菌科,比一般细菌小,能通过细菌滤器,广泛存在于土壤、河流、下水道污泥等环境中。蛭弧菌不侵染真核细胞,对人和动物无致病性~[2-3],薛恒平等~[4-6]报道了蛭弧菌对仔鸡、小鼠、鲫鱼(Carassius auratus)的生长没有毒害作用。蛭弧菌作为"活性抗生素"在农牧业、医学以及环保方面     

乌苏里拟鲿对6种常用渔药的耐受性试验

为了解乌苏里拟鲿对氧化钙、硫酸铜硫酸亚铁合剂(5∶[KG-*1/2]2)、三氯异氰尿酸、敌百虫、聚维酮碘、氯化钠等6种常用渔药的耐受性,在水温28±2 ℃,pH值7.2~8.3,溶氧量4.3~5.6 mg·L-1条件下,将全长3.3~5.8 cm、体质量0.32~1.46 g的当年鱼苗放入不同浓度的上述6种药物的药液中进行耐受性试验。试验结果:聚维酮碘、硫酸铜硫酸亚铁合剂(5∶[KG-*1/2]2)、敌百虫、三氯异氰尿酸、氧化钙、氯化钠对乌苏里拟鲿的96 h半致死浓度依次递减;6种药物对乌苏里拟鲿的安全浓度依次为0.01、0.01、0.06、0.25、7.12和3 347 mg·L-1,均低于防治鱼病常规使用浓度。结果表明,乌苏里拟鲿对氧化钙、硫酸铜硫酸亚铁合剂(5∶[KG-*1/2]2)、三氯异氰尿酸、敌百虫、聚维酮碘的耐受性较弱,应慎用;对氯化钠的耐受性较强,可以用2 %的浓度进行药浴,当水温低于26 ℃,药浴时间少于55 min时尚属安全。     

高效液相色谱-串联质谱法测定池塘沉积物中氯霉素残留

[目的]建立一种使用液相色谱-串联质谱联用仪快速准确检测池塘沉积物中痕量氯霉素残留的方法,为水产品中氯霉素污染来源的排查提供技术支持。[方法]底泥经润湿后,经乙酸乙酯水饱和溶液提取,Sep-Pa K C_(18)固相萃取柱净化,Shim-Pack XR-ODS色谱柱分离,采用甲醇和水为流动相,并用梯度洗脱对待测物进行分离,以氘代氯霉素(氯霉素-D5)为内标,采用负离子扫描,多反应监测模式(MRM),定性离子对m/z为321→152.0,256.8,定量离子对m/z为321→152.0。[结果]氯霉素类药物在0.5~10.0 ng/m L范围内线性良好,相关系数均大于0.99,本方法的检出限为0.1μg/kg,定量限为0.3μg/kg。3个加标水平(0.5、1.0和5.0μg/kg)的平均回收率为90.1%~96.6%,相对标准偏差为2.4%~7.0%。[结论]实验结果符合氯霉素药物残留检测的相关法规要求,适用于淡水池塘沉积物中氯霉素残留的定性定量检测。     

高效液相色谱-串联质谱法同时测定养殖池塘沉积物中喹诺酮类、四环素类和磺胺类抗生素

建立了以高效液相色谱-串联质谱法同时检测养殖池塘沉积物中喹诺酮类、四环素类、磺胺类16种抗生素的分析方法。沉积物样品经提取液[磷酸盐缓冲液(pH=3)∶乙腈=1∶1]提取后,以MAX-HLB串联小柱净化富集,在HPLC/MS/MS多反应监测模式下进行定性及定量分析,当目标化合物添加浓度为10μg/kg和100μg/kg时,沉积物中喹诺酮类、四环素类、磺胺类的加标回收率分别为67.6%~107.1%,63.8%~118.7%,67.1%~116.4%,相对标准偏差在5.1~12.9。以3倍信噪比估算出喹诺酮类、四环素类、磺胺类的检出限分别为0.14~0.64、1.8~3.9、0.18~0.42μg/kg。应用此方法检测21口池塘的43个沉积物样品,结果表明,该方法操作简便,重现性好,可用于池塘沉积物中抗生素的检测。     

寿光蚂蚬滩涂健康养殖技术

正菲律宾蛤仔(Ruditapes philippinarum)是我国四大传统养殖贝类之一,俗称花蛤、蚬子等,多栖息于风浪较小、水流通畅并有淡水注入的内湾中低潮区的泥沙滩涂上~[1-2]。寿光蚂蚬是寿光市别具特色的菲律宾蛤仔品种,由于当地独特的气候、水质与底质等条件,其贝壳厚度和硬度大,肉质结实而富有弹性,口感极佳,深受市场欢迎,更因此成为国家农产品地理标志产品~[3]。寿光市位于渤海莱州湾南畔,     

凡纳滨对虾亲虾性腺成熟节律和交配率

在水温(27±2)℃、盐度29±2、pH 7.8~8.5、光强1 000 lx的条件下,对来自中国台湾省的体质量为35~45 g的凡纳滨对虾(Litopenaeus vannameiBoone 1931)亲虾的性腺成熟节律和交配进行了为期73 d的繁育实验,样品为1 300对,3个批次。结果表明,雌虾术后卵巢成熟率分4期:Ⅰ期(1~9 d)入池适应术后效应期;Ⅱ期(10~16 d)低成熟率期,Ⅲ期(17~51 d)成熟率渐上升期,Ⅳ(52~73 d)平稳期;雄虾精荚形成率分3期:Ⅰ期(1~12 d)入池适应与术后效应期;Ⅱ期(13~51 d)平缓上升期,Ⅲ期(52~73 d)平稳期。在95%的概率保证程度下,雌亲虾卵巢成熟率为9.73%,波动区间9.14%~11.53%;雄虾精荚形成率6.63%,波动区间5.87%~7.38%;平均交配率为61.43%,波动区间58.85%~64.00%。在繁殖过程中,雌虾卵巢再成熟时间间隔9.68 d,可多次成熟的次数19.49次,雄虾精荚再形成时间15.10 d,可多次形成的次数为12.50次。此外还就影响雌虾卵巢成熟和雄虾精荚形成的因素以及如何提高交配率进行了详细的讨论。[中国水产科学,2006,13(4):579-584]     

上海鱼类新记录-方氏鳑鮍

王幼槐[1]1990年在《上海鱼类志》中报道上海地区有3种鳑鮍,即中华鳑鮍Rhodeus sinensisGünther[2]、高体鳑鮍Rhodeus ocellatus(Kner)[3]和彩石鳑鮍Rhodeus lighti(Wu)[4]。经国内外学者研究,彩石鳑鮍为中华鳑鮍的同物异名[5-9]。2006年6月在崇明中兴镇内河采到3 ind鳑鮍属鱼类标本。经鉴定为方氏鳑鮍Rhodeus fangi(M iao)[10],系上海地区鱼类新记录。本种与上述2种的主要区别是体侧中央银蓝色纵纹从尾鳍基向前伸达背鳍起点前方4~5鳞处,而后两种此纵纹不伸达背鳍起点[5-6,11]。现报道如下。方氏鳑鮍Rhodeus fangi(M iao),1934采集时间…     

鱼类脂肪酸结合蛋白研究进展

脂肪酸结合蛋白(FABPs)是一类来自共同祖先基因的小分子蛋白质,相对分子质量为14~16 ku,广泛存在于动物肠道、心脏、脑、脂肪和肌肉等多种组织的细胞内,占细胞内可溶性蛋白总量的1%~8%[1]。研究表明,机体内脂肪酸跨膜运输是被动扩散或由膜蛋白介导的过程,脂肪酸结合蛋白主要结合细胞内游离的脂肪酸(FA)和其他疏水性配体,并将其输送到过氧化物酶体、线粒体和细胞核等细胞器,进而调控脂肪酸氧化和甘油三酯及磷脂合成等脂代谢过程[2](图1)。脂肪酸结合蛋白家族成员的命名是根据其首次分离或鉴定出的组织来命名的,例如肠型脂肪酸结合蛋白(I-FABP)、肝型脂肪酸结合蛋白(L-FABP)等。然而,很多组织中往往有多种脂肪酸结合蛋白基因(fabps基因)同时表达,研究者又对不同的脂肪酸结合蛋白基因按阿拉伯数字命名[3],如fabp1基因(肝型)、fabp2基因(肠型)、fabp3基因(心脏型)等。     

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).     

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.     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.