Cr~（6＋）对泥鳅外周血细胞微核率的影响

利用六价铬离子（Cr6＋）染毒泥鳅（Misgurnus anguillicaudatus）,采用微核试验法,以外周血红细胞的微核率为指标,研究Cr6＋对泥鳅的遗传学毒性。结果显示,Cr6＋对泥鳅具有一定的毒性;当浓度增加或染毒时间延长,均会引起微核率增加,而且高浓度（160～200 mg/L）和长时间染毒（8 d）均出现了显著性差异,微核率与染毒浓度以及与处理时间之间均有较为明显的正相关性,即具有剂量效应和时间效应;研究表明含铬废水排入环境对水生生物会造成不同程度的影响。     

Cr6+对泥鳅外周血细胞微核率的影响

利用六价铬离子(Cr6+)染毒泥鳅(Misgurnus anguillicaudatus),采用微核试验法,以外周血红细胞的微核率为指标,研究Cr6+对泥鳅的遗传学毒性。结果显示,Cr6+对泥鳅具有一定的毒性;当浓度增加或染毒时间延长,均会引起微核率增加,而且高浓度(160～200 mg/L)和长时间染毒(8 d)均出现了显著性差异,微核率与染毒浓度以及与处理时间之间均有较为明显的正相关性,即具有剂量效应和时间效应;研究表明含铬废水排入环境对水生生物会造成不同程度的影响。     

四种除草剂对泥鳅红细胞微核及核异常诱导

采用红细胞微核和核异常测试法,研究了除草剂精禾草克、氟乐灵、扫艹弗特,2-甲-4氯钠水剂对泥鳅红细胞核的遗传毒性。泥鳅在各除草剂试验液中染毒24h,采血制片。结果表明,四种除草剂均不同程度地引起微核细胞率和核异常细胞率等遗传指标的上升,其中部分浓度组较对照组差异显著(P<0 05)或极显著(P<0 01)。除草剂浓度与微核率或核异常率无显著相关,不表现剂量-效应关系。四种除草剂对泥鳅红细胞具有明显的诱变效应,其中扫特的诱变效应要大于其它三种除草剂。     

Cu^2＋对泥鳅外周血红细胞微核及核异常的影响

将泥鳅分别暴露在浓度为0．01、0．05、0．1、0．2、0．4mg／L的Cu^2＋溶液中，常规方法制备血涂片。统计泥鳅外周血红细胞微核率、核异常率。结果显示：不同浓度Cu^2＋均能不同程度地引起泥鳅红细胞傲核率和核异常细胞率等遗传指标的上升，其中，部分浓度组较对照组差异显著（P〈0．05）或极显著（P〈0．01），Cu^2＋浓度和暴露时间与微核率或核异常率无明显剂量-效应关系和时间-效应关系。     

8-羟基喹啉对锦鲤遗传毒性和生理毒性的研究

采用急性毒性,遗传毒性和生理毒性试验,研究了8-羟基喹啉对锦鲤的半致死浓度、红细胞DNA损伤大小及对其肝脏GPT和GOT活性的影响。8-羟基喹啉对锦鲤具有明显的毒性效应,能诱发锦鲤红细胞微核率的增加及降低锦鲤肝组织中GPT和GOT的活性,二者均表现出明显的时间、剂量效应,其中,微核率与处理时间和浓度呈正相关,而酶活性则与处理时间和浓度呈负相关。     

三氯生对红白鲫的急性毒性及遗传毒性研究

研究三氯生对红白鲫的急性毒性和遗传毒性作用,为分析评价三氯生对鱼类的生态安全性提供相应的毒理学资料。试验在规格为58 cm×28 cm×36 cm的玻璃水族箱中进行,红白鲫体长6.8~7.5 cm。采用半静态染毒法,设置不同浓度梯度的三氯生染毒组以及空白对照组和二甲基亚砜溶剂对照组,分别进行为期96 h的急性毒性试验和14 d的遗传毒性试验,观察记录死亡数,统计血红细胞微核率和核异常率。三氯生对红白鲫的96 h LC50为1 111.9μg/L。TCS染毒处理14 d后,红白鲫红细胞核异常的情况明显增多,主要是细胞质中有1~2个微核,其他常见核异常为红细胞核质外凸、核质内凹及核内空泡化;空白对照组和溶剂对照组之间核异常比率无显著差异(P0.05),而三氯生暴露各浓度组(600.0、690.0、793.5μg/L)核异常比率均极显著高于对照组(P0.01),且呈现明显的剂量-效应关系。研究表明,三氯生对红白鲫具有潜在的遗传毒性,且毒性效应在特定条件下随着暴露浓度的增加而增强。     

维生素C对草甘膦诱发的斑马鱼外周血红细胞微核及核异常的缓解作用

研究除草剂草甘膦对斑马鱼(Danio rerio)的急性毒性作用及其对斑马鱼外周血红细胞微核及核异常的影响,同时探讨了维生素C对草甘膦诱发的斑马鱼外周血红细胞微核及核异常的缓解作用。通过急性毒性实验计算出48 h草甘膦对斑马鱼的半致死浓度,通过计数外周血红细胞中微核率和核异常率来分析草甘膦对斑马鱼的遗传毒性;将斑马鱼饲养在含有维生素C的草甘膦水体中48 h,通过观察外周血红细胞中微核率和核异常率的变化情况来分析维生素C对草甘膦诱发的斑马鱼遗传损伤的缓解作用。48 h草甘膦对斑马鱼的半致死浓度为(0.39±0.06)g/L。高浓度草甘膦(0.64~1.28 g/L)对斑马鱼具有极强的致死性,中、低浓度草甘膦(0.16~0.32 g/L)作用斑马鱼48 h后,能显著地诱发斑马鱼外周血红细胞微核和异常核的产生(P<0.05)。在本实验的作用浓度范围内,微核率与核异常率随草甘膦浓度的增加而升高,并呈现明显的剂量-效应关系(P<0.05)。在低浓度草甘膦(0.16 g/L)中加入一定浓度的维生素C(6.25~50 mg/L)后,斑马鱼血细胞中微核率和核异常率可显著降低至清水对照组水平。草甘膦可诱发斑马鱼外周血红细胞微核和核异常的产生,其诱发能力随着草甘膦浓度的增加而增强。在低浓度草甘膦中加入一定浓度的维生素C可以显著降低由其诱发的微核和核异常的产生。     

镉对鲤遗传毒性和外周血细胞数量的影响

将鲤(Cyprinus carpio)分别暴露于0.16、0.31、0.63、1.26 mg/L Cd2+溶液中,研究Cd2+对鲤外周血红细胞微核率、核异常率、红细胞和白细胞数量的影响,探讨Cd2+对鲤遗传毒性和外周血细胞数量影响的变化规律和趋势。结果显示:Cd2+对鲤红细胞微核率和核异常率的影响存在浓度-效应和时间-效应关系。与对照组相比,1.26 mg/L Cd2+组鲤在不同暴露时间的红细胞微核率和核异常率均极显著(P<0.01),核异常率在72 h显著(P<0.05)增加,72 h后表现为极显著(P<0.01)增加。Cd2+对鲤外周血淋巴细胞、单核细胞和嗜中性粒细胞比例的影响存在浓度-效应和时间-效应关系。相同Cd2+浓度下,鲤淋巴细胞比例随暴露时间增加先降低后上升,同一暴露时间淋巴细胞比例随Cd2+浓度增加而降低。相同Cd2+浓度下,鲤单核细胞和嗜中性粒细胞比例随暴露时间增加先升高后降低,同一暴露时间嗜中性粒细胞比例随Cd2+浓度增加而升高。研究表明,Cd2+对鲤具有遗传毒性效应,且随Cd2+浓度增加而增强。     

铜离子对黄鳝血细胞微核及吞噬能力的影响

研究了不同铜离子浓度对黄鳝血红细胞微核率以及血细胞吞噬能力的影响。实验结果表明，铜离子在实验浓度范围内均能诱导黄鳝红细胞微核率，随着铜离子浓度的增高，微核率也明显提高。铜离子还能影响血细胞的吞噬能力，在较低浓度时，吞噬功能增加，但达到一定浓度以后，其吞噬功能开始下降。     

Cd2+对泥鳅外周血红细胞微核及核异常的影响

将泥鳅分别暴露于0.005、0.025、0.05、0.1 mg/L的Cd2+溶液中,常规方法制备血涂片,统计泥鳅外周血红细胞微核率、核异常率.结果显示,Cd2+对泥鳅红细胞微核率的影响具有质量浓度-效应和时间-效应关系.随Cd2+质量浓度的增加,红细胞微核率显著上升.随暴露时间的延长,红细胞微核率呈现增加-降低-增加的趋势.Cd2+对泥鳅红细胞核异常率的影响具有明显的质量浓度-效应关系.随Cd2+质量浓度增加,红细胞核异常率显著上升.随暴露时间的延长,0.1 mg/L组核异常率表现出较为明显的增加-降低-增加的时间-效应关系.     

Cu2+急性毒性下银鲳幼鱼红细胞核异常和抗氧化酶活性的变化

为探究Cu2+对银鲳幼鱼的致毒效应,采用静态毒性实验方法,开展了Cu2+对银鲳幼鱼的急性暴露实验,研究了不同浓度的Cu2+对银鲳幼鱼外周血细胞微核率、核异常率和肝脏抗氧化酶(SOD、CAT、GPX)活性变化的影响。结果表明:Cu2+对银鲳幼鱼48和96 h的LC50分别为0.860和0.770 mg/L,安全质量浓度为0.077 mg/L;Cu2+的浓度与银鲳幼鱼红细胞微核率及核异常率具有明显的剂量效应关系,在相同浓度Cu2+胁迫下,核异常率又普遍高于微核率;不同浓度Cu2+胁迫下银鲳幼鱼肝组织中的SOD、CAT、GPX活性均表现为低浓度被诱导而高浓度受抑制的规律,与Cu2+浓度呈抛物线型剂量效应关系。研究认为红细胞微核率、核异常率与SOD、CAT和GPX活性的变化均可以反映银鲳幼鱼受伤害的程度,并可用作银鲳安全性风险评价的参考依据,其中SOD还可以灵敏地指示低浓度的早期Cu2+污染。     

Determination of histological and genotoxic effects of formalin on Nile tilapia (Oreochromis niloticus L.)

Formalin (37–40% formaldehyde) is one of the most effective, widely used chemical in pisciculture for its antiparasitic, antifungal and prophylactic activities. It is used in paints, cleaning products and textile industry, as well. Genotoxic and histological effects of sublethal formalin exposure on Nile tilapia (Oreochromis niloticus) are investigated in this study. The semi‐static acute test was employed. One‐tenth, one‐fifth and half of the 96‐h LC₅₀ value, 148 mg L^?1, were used as sublethal exposure concentrations: 15, 30 and 75 mg L^?1 for 24, 48, 96 and 168 h respectively. The micronucleus test was applied to investigate the genotoxic effects on fish erythrocytes at the end of predetermined exposure periods and all tissues of the fish were sampled for histological examination. The micronucleus frequencies increased significantly in all exposure groups when compared with their control groups in a dose and time dependent manner (P < 0.05). Results of histological examination showed no histopathological findings in the gonads, heart, skeletal muscle, spleen and intestine tissues after sublethal formalin exposure. Passive hyperaemia of liver tissue in varying proportions, hydropic degeneration, fatty degeneration in high concentrations, branchitis on gill tissues in varying proportions, epithelial lifting, telangiectasia, bleeding in kidney and hyperaemia were found in Nile tilapia exposed to formalin at different concentrations and durations. As a result, it is thought that formalin leads to tissue damage and shows genotoxic effects even at 15 mg L^?1 concentration in O. niloticus.     

苏丹红Ⅲ对泥鳅的急性毒性、遗传毒性及生理毒性研究

以苏丹红Ⅲ为诱变剂,研究了其对泥鳅(Misgurnus anguillicandatus)的急性毒性效应,在此基础上研究了苏丹红Ⅲ在不同浓度和染毒时间对泥鳅红细胞微核率及其肝脏中谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性的影响.试验表明,苏丹红Ⅲ对泥鳅的24h半致死浓度为1.020 g/L,48 h半致死浓度为0.740 g/L,48 h安全浓度为0.117 g/L.染毒一段时间后,泥鳅红细胞的微核率与对照组比较均显著升高,达最高值后趋于平缓;随着苏丹红Ⅲ浓度的升高,泥鳅红细胞微核率与对照组相比出现极显著差异的时间相应提前.染毒后各组的谷草转氨酶和谷丙转氨酶活性呈下降趋势,达到峰值后趋于平缓;染毒时间越长,酶活下降的幅度越大.泥鳅红细胞的微核率及其肝脏中的GOT和GPT活性均与苏丹红Ⅲ有明显的剂量-时间效应,研究结果为进一步明确苏丹红Ⅲ的危害提供了试验数据.     

Efficacy of an equal blend of canola oil and poultry fat as an alternate dietary lipid source for Atlantic salmon (Salmo salar L.) in seawater. II: effects on haematology and immunocompetence

This research examined the haematological and immunological responses of quadruplicate groups of juvenile (～400 g initial weight) Atlantic salmon (Salmo salar L.) that had each been fed daily to satiation for 12 weeks one of three high‐energy extruded diets of identical composition except for the supplemental dietary lipid (234.7 g kg^?1) source. The three experimental diets varied in the composition of supplement lipid; diet 1 contained 100% anchovy oil (AO), while diets 2 and 3 replaced 29.8% and 59.7% of the AO (respectively) with a 1:1 blend of canola oil (CO) and poultry fat (PF). Immediately following the feeding trial, a random sample of fish from each diet was sampled for determination of baseline levels of various haematological and immunological parameters. Thereafter, duplicate diet groups were vaccinated (against Listonella anguillarum) and reared on their respective experimental diets for an additional 4 weeks. At that time, the remaining fish were sampled similarly, and the different parameters were measured again. Comparisons between the different diet treatment groups were made before and after vaccination. There were no significant diet treatment effects at either sample time, for haematocrit, differential leucocyte counts, erythrocyte counts, serum hemolytic activity or head kidney leucocyte respiratory burst activity. The fish fed diet 1 however, did show significantly higher post‐vaccination levels of peripheral blood leucocyte respiratory burst activity and higher serum antibody titres against L. anguillarum. The results suggest that the relatively low n‐6/n‐3 fatty acid ratios in the muscle and presumably other tissues of fish fed diet 1, may have resulted in a reduced production of immunocompromising eicosanoids than were produced in fish ingesting the other two diets that were based in part on the different amounts of the CO and PF blend. Long‐term studies are required to confirm this possibility.     

A comparison between non-destructive and destructive testing of Atlantic salmon, Salmo salar L., broodfish for IPNV--destructive testing is still the best at time of maturation.

Two populations of Atlantic salmon broodstock, previously identified as infectious pancreatic necrosis virus (IPNV) carriers, were screened for IPNV at the time of stripping. Four hundred and ten broodfish were individually sampled of which 91 were detected as IPNV positive by virus culture of sonicated kidney homogenates combined with gonadal fluid, but none tested positive by the blood leucocyte assay. Thirty fish identified as IPNV carriers prior to maturation by the blood leucocyte assay were used in a separate study to compare non-destructive vs. destructive testing methods at stripping. IPNV was not detected using the blood leucocyte method at the time of stripping. RT-PCR and real-time PCR assays failed to detect IPNV from 13 blood samples, the virus was not isolated from milt (0/14) or sonicated ovarian fluid cell pellets (0/16) and only three fish tested positive by the standard culture of kidney homogenates. A third study of Atlantic salmon broodfish compared the IPNV isolation rates prior to maturation with the isolation rates at spawning during 1999-2001. In each year the percentage of IPNV-positive broodfish was significantly lower than in the pre-broodstock sample. While in pre-broodfish samples IPNV was detected by the blood leucocyte assay, no culture isolations or PCR positives were detected from non-destructive samples of the same individual broodfish at stripping. A consistent finding was that even for the kidney assay, the percentage of IPNV-positive fish in carrier populations was higher in pre-broodstock than in broodfish at stripping. These results indicate that destructive kidney sampling is still the most sensitive method for detecting IPNV carrier Atlantic salmon broodfish and that a change in IPNV carrier-status occurs during the maturation period.     

注射菌苗的生殖期黄颡鱼亲鱼的外周血细胞和体液免疫变化

阐明生殖期亲鱼在注射灭活菌苗后的血细胞和体液免疫变化规律，是预防亲鱼产后虚弱患病和仔鱼染病死亡的理论基础。本研究在试验组和对照组黄颡鱼胸鳍基部分别注射0.2 mL浓度为1.0×10⁸ CFU/mL福尔马林灭活温和气单胞菌苗（F-AS）和0.65%灭菌生理盐水，在免疫后第1、2、4、7、14、21、28、35天，从尾静脉采血，测定外周血液血细胞数量及组成比例，血细胞吞噬活性和抗体效价。结果显示：注射菌苗后白细胞数量和淋巴细胞的白细胞分类计数逐渐增加，显著高于对照；中性粒细胞、单核细胞的白细胞分类计数以及吞噬百分比（PP）、吞噬指数（PI）在4～7 d显著高于对照；抗体效价逐渐升高且在第14天达到最高，随后逐渐下降；而红细胞数和血栓细胞的白细胞分类计数逐渐下降，显著低于对照组。结果表明：单次注射菌苗后亲鱼可以产生良好的免疫应答，1～7 d以增强单核细胞和中性粒细胞等非特异细胞免疫为主；7～35 d淋巴细胞增值，并大量释放抗体且在第21天淋巴细胞出现极显著升高，此后虽逐渐下降但仍显著高于对照，抗体维持在较高水平上的时间为1周左右，建议在黄颡鱼催产前10～14 d进行单次菌苗注射以增强亲鱼免疫力。     

Effects of dietary tryptophan levels and fish stocking density on immunological and antioxidant responses and bactericidal activity against Aeromonas hydrophila in rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss) was reared under low (LD) and high (HD) stocking densities for 70 days, during which they were fed with diets supplemented with 0, 5 and 10 g tryptophan (Trp) per kg. At the end of the experiment, there were significant interaction effects of the stocking density and Trp levels on plasma Trp, globulin, lysozyme, catalase (CAT), malondialdehyde (MDA), and bactericidal activity and blood leucocyte count. However, there was no difference in plasma complement (ACH50), total immunoglobulin (Ig), total antioxidant capacity (TAC), superoxide dismutase (SOD) and blood differential leucocyte count. Trp at 5 g per kg diet significantly increased lysozyme and bactericidal activity, and decreased MDA under both the LD and HD conditions. Moreover, it significantly increased plasma CAT and blood leucocyte under the LD and HD conditions respectively. Trp at 10 g per kg diet significantly increased blood leucocyte and plasma bactericidal activity under the LD condition, but significantly decreased plasma globulin and bactericidal activity under the HD condition. In conclusion, the present results showed that the effects of Trp on immune and antioxidant systems depend on Trp levels and stressful conditions. As 5 g Trp per kg diet is beneficial for rainbow trout well‐being under both the LD and HD conditions, higher level of Trp (10 g/kg) is not beneficial and even causes some negative effects under the HD condition.     

Effects of substitution of dietary fish oil with a blend of vegetable oils on liver and peripheral blood leucocyte fatty acid composition, plasma prostaglandin E2 and immune parameters in three strains of Atlantic salmon (Salmo salar)

Duplicate groups of three genetic strains of Atlantic salmon smolts were cultured on diets containing either fish oil (FO) or a blend of vegetable oils (VO). Fatty acid compositions of liver and peripheral blood leucocytes were compared. The effect of different strains and diets on innate immune parameters and plasma prostaglandin E₂ were also measured. Two strains were selected as being either 'fat' or 'lean' in terms of muscle adiposity. The third strain was a commercial stock (MH). Total replacement of dietary FO with VO resulted in reduced docosahexaenoic (DHA; 22:6 n -3) and eicosapentaenoic acids (EPA; 20:5 n -3) in liver, while oleic (18:1 n -9), linoleic (18:2 n -6) and α-linolenic (18:3 n -3) acids were all increased in VO-fed fish. Fatty acid compositions of blood leucocytes showed similar changes. Evaluation of innate immune function showed that in the fat strain, circulating leucocytes were significantly lower in VO fish. The lean strain also had significantly higher serum lysozyme activity than MH fish. Reduced haematocrit was seen in VO lean fish compared with FO lean fish. This study provides evidence of strain-induced differences in liver and leucocyte fatty acid compositions and innate immunity in Atlantic salmon fed either FO- or VO-based diets.