泥蚶血清及血细胞免疫功能研究

初步研究了泥蚶(Tegillarca granosa)血细胞离体条件下的吞噬作用以及血清因子对其的影响,并测定经大肠杆菌注射以后,泥蚶血清中抗菌、溶菌、酚氧化酶(PO)、超氧化物歧化酶(SOD)活力的变化。结果表明,泥蚶血清能促进血细胞对大肠杆菌的吞噬,但不能识别鸡红细胞。大肠杆菌激发后泥蚶血清中抗菌活力在1h、15h和30h时实验组均显著高于对照组。溶菌活力实验组均高于对照组,但在15h差异不显著。PO活力实验组均低于对照组,在30h时差异极显著。SOD活力在1h时显著低于对照组,而在30h时显著高于对照组。     

食盐对草鱼血清超氧化物歧化酶和溶菌酶活性的影响

草鱼养殖过程中经常使用食盐药浴,以便在高渗环境下杀死病原体。研究食盐药浴后草鱼代谢酶变化,有助于了解食盐药浴对草鱼机体生理免疫的影响。将草鱼（Ｃｔｅｎｐｈａｒｙｎｇｏｄｏｎｉｄｅｌｌａ）分别暴露于食盐浓度为０、２％、３％、４％水体中浸浴１０ｍｉｎ,然后置入水簇箱中饲养,分别在药浴后１２ｈ、２４ｈ、３６ｈ取样,测定其血清中超氧化物歧化酶（ＳＯＤ）和溶菌酶（ＬＳＺ）活性变化。结果显示,与对照组相比,血清ＳＯＤ活性在１２ｈ时,２％组中显著上升（Ｐ＜０．０５）,３％、４％组显著下降（Ｐ＜０．０５）;２４ｈ取样时,２％、３％组中无显著差异（Ｐ＞０．０５）,４％组显著下降（Ｐ＜０．０５）;３６ｈ取样时,各组间均无显著差异（Ｐ＞０．０５）;血清ＬＳＺ活性各组间均无显著差异（Ｐ＞０．０５）。食盐能引起草鱼ＳＯＤ活性变化,而对溶菌酶活性影响不大;表明食盐对草鱼不同代谢酶有不同的反应模式,抗氧化酶ＳＯＤ对食盐应激有敏感的生理应答。     

泥蚶血细胞耐饥饿及抗菌力特性的研究

显微镜下观察泥蚶血细胞抗菌能力以及在不同饥饿条件下细胞形态结构的变化。结果表明:经饥饿处理后,泥蚶血细胞随着时间的迁移,出现细胞形态不规则、胞质色泽变浅、胞质颗粒物减少、胞膜边缘光滑度减小等现象。血细胞对大肠杆菌、奥斯陆莫拉氏菌、脲放线杆菌均具较高的噬菌率,噬菌实验过程中血细胞形态结构以及细胞组分不断产生变化。细胞抗菌活性检测结果表明:血细胞对三者都具有一定的抗菌活性,活性依次为:大肠杆菌>奥斯陆莫拉氏菌>脲放线杆菌。     

微生态制剂对提高杂交鲤越冬能力的研究↑（*）

利用微生态制剂（Ｊｙ１０节杆菌ＡｒｔｈｍｏｂａｃｔｅｒＳＰ，Ｊｙ３１乳杆菌Ｌａｔｏｂａｃｉｌｕｓｃａ－ｓｅｉ制成的活菌制剂）饲养并定期灌喂鲤，３０－３５天后在低温条件下感染致病菌，观察死亡率、发病情况和测定各项生理生化指标。结果表明，微生态制剂实验组死亡率明显低于对照组，半数致死量（ＬＤ５０）１０－０．７９９高于对照组１０－０．９２１，差异显著（Ｐ＜０．０５）；血清总蛋白、ｒ－球蛋白、血糖量、血脂量、红细胞脆性、脑胆碱酯酶活力及白细胞吞噬功能等，实验组均优于对照组     

细菌糖蛋白对螯虾免疫因子的影响

实验检测了气味黄杆菌的胞外糖蛋白对克氏原螯虾（Ｐｒｏｃａｍｂａｒｕｓ ｃｌａｒｋｉｉ）的几咱非特异性免疫因子－凝集素、酚氧化酶、溶菌、抗菌和超氧化物歧化酶等活力的影响。注射１０ｍｇ糖蛋白后,实验组敖蛳主血清的免疫因子活力均高于对照组,凝集素活力在第１～４天最高;酚氧化酶在第１～５天维持较高活力,最大活力在第２天,溶菌、抗菌活力在第１～６天维持较高水平,分别在第３天、第２天达到最大。与对照组相比,实     

双齿围沙蚕与赤子爱胜蚓对凡纳滨对虾生长和免疫指标的影响

以鲜活饵料双齿围沙蚕（Perinereis aibuhitensis）、赤子爱胜蚓（Eisenia foetida）和人工配合饵料投喂凡纳滨对虾（Litopenaeus vannamei），并测定各种饵料对其生长和免疫指标的影响。结果表明，沙蚕和蚯蚓单独投喂或与人工配合饵料配合投喂都可显著提高凡纳滨对虾的生长速率，但用沙蚕单独投喂的对虾成活率较低。蚯蚓与人工配合饵料配合投喂可显著提高对虾血清中抗菌活力、溶菌活力、酚氧化酶活力和血细胞数量；蚯蚓单独投喂可显著提高血清中抗菌活力、酚氧化酶活力和血细胞数量，对血清中溶菌活力作用不明显。沙蚕与人工配合饵料配合投喂可显著提高对虾血清中抗菌和溶菌活力．但对血清中酚氧化酶活力和血细胞数量变化作用不明显；单独投喂沙蚕对所测各项免疫指标影响均不显著。研究结果证实：利用蚯蚓和沙蚕可显著提高对虾的生长速率，但蚯蚓的饵料效果优于沙蚕。[中国水产科学，2006，13（4）：561—565]     

聚甘露糖醛酸对中国对虾免疫相关酶活性和溶菌溶血活性的影响

在中国对虾的血清、肌肉和肝胰提取液中有酸性磷酸酶（ＡＣＰ）、碱性磷酸酶（ＡＬＰ）、过氧化物酶（ＰＯＤ）,其中,ＡＣＰ和ＡＬＰ在肝脏中的活性最高。采用１．０％的聚甘露糖醛多糖作为免疫药物,对中国对虾进行注射刺激后,发现血清中的溶菌和溶血活性有显著提高;肝胰腺中的ＡＣＰ和ＡＬＰ活性也有明显增加,在７２ｈ,ＡＣＰ活性由对照组的３．２１Ｕ．１００ｍＬ＾－１提高到９．６９Ｕ．１００ｍｌ＾－１,ＡＬＰ活性由４     

病毒感染对栉孔扇贝血淋巴中过氧化氢酶和过氧化物酶活力的影响

用从病贝体内分离的非包涵体球形病毒感染栉孔扇贝,采用分光光度技术,分别于6,24,72h,测定栉孔扇贝血清和血细胞中过氧化氢酶(CAT)活力和过氧化物酶(POD)活力。结果表明,感染病毒后,血清中CAT活力在6,24,72h时,实验组均高于对照组,6h时差异极显著,24h时差异显著,72h时差异不显著;血细胞中CAT活力在6,24,72h时,实验组均高于对照组,6h和24h时差异极显著,72h时差异不显著。血清和血细胞中POD活力在6h和24h时,实验组均高于对照组,且均差异极显著。说明病毒感染后栉孔扇贝血淋巴中CAT和POD的活力增强。     

插核手术对合浦珠母贝免疫水平的影响

对插核手术前后合浦珠母贝血清的超氧化物岐化酶（SOD）活力、溶菌活力和抗菌活力以及血细胞的吞噬活性进行了比较研究。结果表明：插核手术后贝血清的SOD活力明显上升,溶菌、抗菌活力明显下降,血红细胞的吞噬活性降低,化学发光的各种参数显著改变,说明贝的体液和细胞免疫水平都明显降低。据此推断手术贝的免疫力低下是导致手术贝高死亡率的最根本原因。     

鱼鳞提取物延缓衰老作用研究

报告了鱼鳞提取物对所丰年大鼠血清,肝脏丙二醛含量和超氧化物歧化酶活力的影响。结果表明,实验组大鼠血清,肝脏脂质过氧化物丙二醛含量显著低于正常对照组,实验组大鼠血清,肝脏ＳＯＤ活力增加。对尾腱及皮肤的氨基酸分析显示,实验组大鼠尾腱皮肤中羟脯氨酸含量显著高于正常对照组,提示鱼鳞提取取得具有延缓衰老的作用。     

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.