急性拥挤胁迫对鲤血液中皮质醇及几项生化指标的影响

通过对鲤(Cyprinus carpio)3 h急性拥挤胁迫(密度70g/L)实验,并在胁迫开始前(对照)、胁迫结束后的第0至168 h采样测量几项血液指标。实验结果显示在3 h拥挤胁迫结束之时(0 h),鲤血清皮质醇、血糖和乳酸水平均显著高于胁迫前对照值(P<0.05),而游离脂肪酸水平极显著低于对照值(P<0.01)。之后这些指标继续发生波动,但一段时间后均可恢复至对照水平并保持稳定。实验期间血清甘油三酯和胆固醇水平并未出现显著变化(P>0.05)。该结果提示3 h拥挤胁迫可引起鲤体内血清皮质醇水平及部分血液生化指标发生改变,这可能是为了弥补应激期间机体对能量需求的增大。     

运输胁迫对银鲳血清皮质醇、血糖、组织中糖元及乳酸含量的影响

研究分析了运输胁迫2、4、8和12 h后银鲳血清皮质醇、血糖、组织中糖元及乳酸含量的变化。选取平均体质量为(10.05±0.23) g的银鲳幼鱼为试验对象,共设3组运输密度,分别为4、8和16 g/L。每个时间段的运输结束后,将取样后剩余的鱼转入1 m³水体的圆形玻璃钢桶中观察24 h,以统计各自的累计死亡率。研究结果显示,16 g/L密度组在运输12 h后成活率为91.25%,其余均为100%;运输胁迫导致皮质醇含量显著升高(P<0.05),运输4 h后,血清皮质醇含量均达到最高值,12 h后皮质醇含量虽较运输4 h后有所降低,但仍显著高于试验初的水平,且此时16 g/L密度组的皮质醇含量均显著高于4和8 g/L密度组(P<0.05);血糖、血清与肌肉中乳酸含量在运输胁迫后均显著升高(P<0.05);肝脏与肌肉中糖元含量在运输胁迫后均显著降低(P<0.05),但在整个运输胁迫过程中,肝脏组织中糖元含量的降低程度明显高于肌肉组织。统计分析得出,较高的运输密度会加重银鲳的应激性反应,并推测在小水体开放式运输的情况下,规格为10 g左右的银鲳幼鱼,其运输密度不宜超过16 g/L。此外,在运输胁迫下,银鲳机体的能量供给主要来自肝脏糖元的动员。     

不同水平维生素E对高温应激及常温恢复后团头鲂血清生化指标、肠道抗氧化能力的影响

为研究饲料中添加不同水平的维生素E(VE)对急性热应激及常温恢复条件下团头鲂血清生理生化、激素及肠道抗氧化指标的影响,实验挑选180尾健康、规格一致的团头鲂(45.00±0.50)g,分成3组,每组4个平行,每组分别添加不同量维生素E(0、50和400 mg/kg饲料)。饲养90 d后,对团头鲂进行32℃应激1 d,并在25℃常温恢复7 d。结果表明:应激前,50 mg/kg VE组显著提高了肠道谷胱甘肽的活性(P<0.05);高温应激后,与对照组相比,50 mg/kg VE组在高温应激1 d后显著降低了谷丙转氨酶和皮质醇含量,常温恢复7 d后显著降低了谷草转氨酶、皮质醇的含量(P<0.05),显著提高了碱性磷酸酶、谷胱甘肽和超氧化物歧化酶活性(P<0.05);与对照组比,400 mg/kg VE添加组在32℃应激1 d及25℃恢复1 d后显著降低了谷丙转氨酶活性(P<0.05),常温恢复7 d后显著降低了皮质醇浓度,在应激1 d及恢复7 d后均显著降低了丙二醛浓度(P<0.05),400 mg/kg VE添加组在高温应激1 d后显著提高了碱性磷酸酶活性和三碘甲腺原氨酸含量,常温恢复7 d后显著提高了碱性磷酸酶活性(P<0.05)。因此,添加适量的维生素E(50～400 mg/kg)有利于调节血脂变化,提高团头鲂肠道抗氧化能力,并且可缓解高温应激对团头鲂血液指标波动的影响,减轻脂质过氧化水平,对团头鲂起到一定保护作用。     

船舶噪声对大西洋鲑体质量、皮质醇及部分免疫指标的影响

为探讨船舶噪声对大西洋鲑(Salmo salar)体质量、皮质醇及部分免疫指标的影响,选取平均体质量为(44.01±11.0)g的大西洋鲑,通过噪声处理组[(120±5)dB、(150±5)dB]以及对照组试验,测定大西洋鲑体质量以及机体血清中皮质醇激素(ORT)、碱性磷酸酶(AKP)及总胆红素含量的变化情况。结果显示:试验组和对照组的大西洋鲑体质量增长差异不显著(P>0.05);血清中总胆红素、碱性磷酸酶、皮质醇从试验开始至结束,差异显著(P<0.05);在第10天,试验组的质量浓度显著高于对照组,呈现先上升后下降并趋于稳定的趋势。两试验组存在差异显著(P<0.05),各生化指标质量浓度呈现先上升后下降并趋于稳定的趋势。该研究可为大西洋鲑工业化健康养殖提供参考。     

丁香酚对“优鲈3号”幼鱼运输水质及其血液、肌肉生理指标的影响

为探索适合“优鲈3号”幼鱼的麻醉运输方法,开展了丁香酚对“优鲈3号”幼鱼的静水麻醉试验和模拟麻醉运输试验,监测了麻醉运输试验中的水质变化和幼鱼血液、肌肉生理指标的变化。结果表明：随着麻醉剂浓度的升高,幼鱼进入麻醉的时间缩短,复苏时间延长,静水麻醉使用丁香酚的最适质量浓度为12～18 mg/L。综合考虑运输需求、幼鱼存活率以及残留代谢,选择能达到轻度麻醉期的6 mg/L为模拟麻醉运输试验的麻醉剂浓度。以质量浓度为6 mg/L的丁香酚对幼鱼进行麻醉后进行10 h模拟运输,分析运输水质、幼鱼生理指标的变化发现：随着运输时间的延长,麻醉组和对照组水体的pH均显著降低,麻醉组的pH总体高于对照组;随着运输时间的延长,水体中氨氮质量浓度显著升高,麻醉组总体低于对照组。幼鱼血清中的皮质醇(COR)浓度随着运输时间的延长呈现先升高再下降的趋势,且麻醉组峰值出现的时间早于对照组。幼鱼血清中尿素氮(BUN)和乳酸(LD)含量以及谷草转氨酶(AST)活性也随着运输时间的延长而升高,麻醉组总体低于对照组。幼鱼肌肉中的乳酸与pH呈负相关关系,乳酸含量随着运输时间的延长而显著升高,pH则总体呈下降趋势。     

热应激对大正三色锦鲤非特异性免疫指标及HSP70基因表达的影响

为探讨锦鲤的抗逆机理,在实验室条件下研究了持续热应激对大正三色锦鲤非特异性免疫指标及HSP70基因相对表达量的影响。分别于应激前、应激后2、6、10、14、18、22、26 h进行取样测定呼吸爆发、补体蛋白3(C3)、超氧化物歧化酶(SOD)、丙二醛(MDA)以及热休克蛋白70(HSP70)基因的相对表达量。结果发现,热应激后呼吸爆发降低,在应激后2、10、18～26h降低显著(P<0.05);热应激2 h后血清中C3含量略有上升,应激6～26 h过程中与应激前相比均下降,应激14～22 h显著地下降(P<0.05);热应激下血清中SOD有上升的趋势,但差异不显著(P>0.05);热应激下,血清中MDA浓度升高,应激10～18 h上升显著(P<0.05);应激后2、26 h HSP70基因的相对表达量与应激前相比显著性地上升了5.93倍、2倍(P<0.05),应激后6～22 h HSP70基因的相对表达量与应激前水平无显著性差异(P>0.05)。结果表明,热应激影响锦鲤非特性免疫指标,降低锦鲤非特异性免疫力;HSP70的表达受热应激调节,热应激下诱导合成的HSP70对锦鲤起到一定的应激保护作用。     

两种麻醉剂对罗非鱼携氧相关血液指标的影响

分别用3个浓度梯度的MS-222和苯唑卡因对尼罗罗非鱼(Oreochromis niloticus)(6.050 0±1.710 3)g进行6 h浸泡式麻醉(水温30.7~32.3℃),然后抽取尾部静脉血,测定血液生理生化指标。重点分析了与生物携氧功能密切相关的6个指标的变化,结果显示:(1)较高浓度(25 mg/L、40 mg/L)MS-222导致红细胞数目(Er)升高,而苯唑卡因导致该值大幅下降;(2)两种麻醉剂对红细胞平均血红蛋白含量(MCH)都无显著影响,且MS-222处理后,血液的血红蛋白浓度(Hb)也无明显变化,但经苯唑卡因麻醉处理后,Hb显著下降,且降幅随药物浓度升高而显著增大。     

运输对养殖黄颡鱼血液生化和肌肉物性分析指标的影响

测定了在10.4~11.2℃下运输4 h后不同时间(0 h、1 h、3 h、6 h、12 h、1 d、3 d、7 d和20 d)养殖黄颡鱼(Pelteobagrus fulvidraco)血液生理生化指标和肌肉物性分析指标的变化。结果显示,运输引起了黄颡鱼血清皮质醇和血糖浓度的显著升高(P0.05),在运输结束12 h后恢复到正常水平。运输后鱼体血清超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和总抗氧化能力(T-AOC)均有显著性升高,随着恢复时间的推移逐渐回降到正常水平;但过氧化物酶(POD)活性并未出现显著性差异,维持稳定的水平。运输后肌肉硬度、弹性、胶黏性、凝聚性、咀嚼性、回复性等物性分析指标均显著性降低。除了凝聚性和咀嚼性之外,其他肌肉物性分析指标在运输结束72 h后恢复到正常水平。研究表明,长途运输造成了养殖黄颡鱼的应激反应,对肌肉物性特征产生了显著的影响,但运输后的恢复可使鱼体抗氧化能力和肌肉物性分析指标回复到正常水平。     

热应激对银鲫生化指标和HSP70 mRNA表达的影响

本研究旨在探讨热应激对银鲫(Carassius auratus gibelio)生化指标及肝脏HSP70 mRNA表达的影响,选取平均体重为(72.8±10.1) g的银鲫,分别在热应激前(25℃,对照组)、热应激后(32℃,0 h、2 h、6 h、10 h)和恢复适温后6 h (25℃,恢复组)的条件下,测定机体血浆乳酸(LD)、血浆葡萄糖(Glu)、肝糖原(Gly)、己糖激酶(HK)、丙酮酸激酶(PK)和肝脏HSP70 mRNA表达量变化。研究表明,实验组乳酸含量从热应激开始至恢复期,差异始终不显著(P>0.05);实验组血浆葡萄糖水平在热应激后0 h、2 h、6 h,极显著高于对照组(P<0.01);实验组肝糖原水平在热应激0 h时,极显著高于对照组(P<0.01),其余时间点差异不显著(P>0.05);己糖激酶活力在热应激后2 h,显著高于对照组(P>0.05);丙酮酸激酶活力在热应激后6 h,极显著高于对照组(P<0.01);热应激后2 h,肝脏中HSP70 mRNA表达量达到最高(P<0.01),恢复至适温后,表达量与应激前无差异(P>0.05)。本研究为进一步探究银鲫的高温应激作用机制及调控机理提供参考数据,同时,也为银鲫养殖和育种等生产实践提供相应参考依据。     

MS-222和丁香油对黄条鰤麻醉效果研究

黄条鰤(Seriola aureovittata)是一种全球水域广泛分布的具有长距离洄游特性的大型大洋性鱼类,其生性凶猛、游泳速度快,人工养殖条件下对环境刺激的应激反应强烈。为开发适宜黄条鰤实验和生产操作的低应激处理技术,采用实验生态学、生理学手段,首次研究了2种温度条件下(20℃和24℃)、2种麻醉剂(MS-222和丁香油)对1龄黄条鰤的麻醉效果,通过行为判别与生理指标分析(皮质醇、肾上腺素和葡萄糖的水平),确定了2种麻醉剂对黄条 的最适麻醉时间、复苏时间和麻醉剂量。根据黄条鰤麻醉和复苏时的行为特征,将麻醉过程分为7个时期,复苏过程分为6个时期。在水温为20℃和24℃时,MS-222麻醉黄条 的最佳浓度分别为100~120 和100 mg/L,而丁香油的最佳浓度均为40 mg/L。在此条件下,实验鱼均可在3 min之内入麻,5 min之内复苏。随着麻醉剂浓度的增加,黄条鰤入麻时间呈缩短趋势,复苏时间呈延长趋势。水温的升高降低了入麻时间,而对复苏时间无明显影响。在水温20℃和24℃条件下,40 mg/L的丁香油麻醉黄条鰤后,血清皮质醇水平分别于6 h和12 h达峰值;在水温24℃条件下,100 mg/L的MS-222麻醉黄条鰤后,血清肾上腺素水平于12 h达峰值,其他各实验组血清肾上腺素、葡萄糖和皮质醇水平均在24 h达峰值。2种温度条件下,实验鱼血清中3种应激相关生理指标在复苏72 h后均显著降低至初始水平以下。本研究结果可为制定规范化的黄条鰤实验与养殖生产操作技术提供参考依据。     

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.