铜在鲤体内的蓄积及毒性的研究

采用半静态法将鲤(Cyprinus carpio)暴露于不同浓度的铜溶液中,研究了铜在鱼体中的吸收富集及毒性。结果表明:铜在鲤肝、肾、鳃和肌肉中的积累量均随着铜浓度的升高而增加。鲤各部位对铜的蓄积能力依次是肝、肾、鳃、肌肉,且肝、肾中铜的积累量明显高于鳃和肌肉,鲤对铜具有较强的蓄积毒性。中毒鱼体色变黑,生长受到抑制,临死前出现神经症状;鳃小片上皮增生、变性、坏死;肝细胞和肾小管上皮细胞空泡变性、溶解性坏死。     

鲤慢性喹乙醇中毒的病理学和组织残留

试验以含10、25、50、100、150和200mg·kg-1喹乙醇的饲料对鲤进行了慢性毒性试验。经90d的试验,各组的发病率分别为17.5%、27.5%、35.0%、42.5%、47.5%和55.0%,死亡率分别为5.0%、10.0%、12.5%、17.5%、20%和30.0%。中毒鱼表现为特征性的"应激性出血综合征",且Hb含量和RBC数量降低,血清AST、ALT活性升高。病理组织学表现为鳃小片水肿,上皮细胞增生、变性、坏死和脱落。心肌纤维、肾小管上皮细胞和肾上腺细胞空泡变性;肝细胞脂肪变性或水泡变性及溶解坏死;肠道发生卡他性肠炎;超微结构上,心肌纤维、肝细胞和肾小管上皮细胞的线粒体肿胀、嵴断裂、溶解,粗面内质网扩张,核糖体脱颗粒;肝细胞内脂滴明显增多;肠上皮微绒毛脱落、崩解和肠上皮细胞大量坏死脱落。中毒鱼肝、肾和肌肉组织内喹乙醇含量与对照组之间有显著性差异。     

鲮鱼肝细胞超微结构研究

应用透射电镜对鲮鱼肝脏细胞进行了观察。结果表明, 正常状态下肝细胞具有典型的单核现象, 核仁、核膜、核孔等都清晰可见, 肝细胞含有线粒体、粗面内质网、溶酶体、糖原等细胞器和内容物, 胆小管主要为胞内胆小管。饥饿4周后, 鲮鱼肝细胞所含的细胞器总数目减少, 核周粗面内质网含量降低, 线粒体出现多种形状, 其间电子密度较低, 嵴更加稀疏, 溶解, 无全嵴, 其最大变化是肝细胞内的糖原几乎消失, 出现大量的溶酶体。再恢复投喂4周后, 鲮鱼肝细胞各种细胞器均恢复到正常水平, 鲮鱼的糖原大量出现, 占据细胞大量空间。表明鲮鱼肝细胞代谢与细胞器数目和内容物密切相关, 在一段时间的饥饿胁迫之后, 其肝脏能过量地贮存糖原作为能源物质。     

鲮鱼肝细胞超微结构研究

应用透射电镜对鲮鱼肝脏细胞进行了观察.结果表明,正常状态下肝细胞具有典型的单核现象,核仁、核膜、核孔等都清晰可见,肝细胞含有线粒体、粗面内质网、溶酶体、糖原等细胞器和内容物,胆小管主要为胞内胆小管.饥饿4周后,鲮鱼肝细胞所含的细胞器总数目减少,核周粗面内质网含量降低,线粒体出现多种形状,其间电子密度较低,嵴更加稀疏,溶解,无全嵴,其最大变化是肝细胞内的糖原几乎消失,出现大量的溶酶体.再恢复投喂4周后,鲮鱼肝细胞各种细胞器均恢复到正常水平,鲮鱼的糖原大量出现,占据细胞大量空间.表明鲮鱼肝细胞代谢与细胞器数目和内容物密切相关,在一段时间的饥饿胁迫之后,其肝脏能过量地贮存糖原作为能源物质.     

急性氨氮胁迫及毒后恢复对团头鲂幼鱼鳃、肝和肾组织结构的影响

为进一步了解氨氮对团头鲂幼鱼的毒性毒理影响,以体质量为(14.27±0.01)g的团头鲂幼鱼为研究对象,研究了氨氮胁迫对其鳃、肝、肾组织结构的影响。实验首先进行96h的氨氮胁迫,得出96h LC50,在此基础上,设置对照组(0.472 mg/L)和实验组(25 mg/L)两个氨氮浓度处理组,进行0、6、12、24、48h的氨氮胁迫,取样后剩余团头鲂幼鱼移入曝气自来水进行96h的毒后恢复实验。结果表明：96h LC50为56.492 mg/L;三种组织观察表明,氨氮胁迫6h,鳃丝毛细血管扩张,上皮组织增生;肝细胞肿胀,细胞核肿大,肝细胞空泡化;肾小球萎缩,肾小囊腔膨大,肾小管管腔缩小;胁迫12h,泌氯细胞增生,呼吸上皮细胞出现部分脱落;肝细胞水样变性、血窦扩张、细胞轮廓模糊,形成点状病灶;肾小管上皮细胞肿大、水样变性、浊肿;胁迫24h,鳃小片融合、变短,呼吸上皮细胞大面积脱落;肝细胞水样变性、血窦扩张严重,形成局部病灶;肾组织淋巴细胞浸润严重,充血,肾小球坏死,肾小管坏死;胁迫48h,鳃小片卷曲,上皮细胞部分脱落;肝细胞部分溶解、血窦扩张,形成点状病灶;肾小管上皮细胞坏死,肾小球坏死;96h恢复后,泌氯细胞和上皮组织增生严重;肝组织大面积细胞核肿大,血窦扩张;肾组织淋巴细胞浸润严重,肾小管坏死,肾小球坏死。实验表明,不同的器官之间病症的损伤程度是不同的,肝组织的损伤最严重,然后依次是鳃和肾。随着胁迫时间延长,鳃、肝和肾组织受到的损害增加,同时鱼体也产生防御反应,但96h的恢复期不足以让团头鲂幼鱼在胁迫中完全恢复,而恢复能力最差的是肾组织。     

Mo~(6+)对草鱼组织器官及抗氧化酶活性的影响

通过不同质量浓度Mo~(6+)(0、30.67、122.66mg/L)对(10.0±3.0)g草鱼幼鱼脑、肝脏、肾脏、心脏及性腺的组织学以及对肝胰脏抗氧化酶活性的影响,探讨了重金属Mo~(6+)的毒性积累和毒性机制。结果表明,草鱼Mo~(6+)中毒后,脑组织出现异常,表现为细胞破裂、细胞核聚集程度严重;肾脏充血,肾小管上皮细胞核肥大,肾小球明显萎缩;心脏肌纤维细胞体积增大,单位视野内细胞数变少,细胞核形态异常,以及局部的细胞空泡变性,出现少量细胞核偏移和密集并且细胞轮廓模糊,肌纤维有轻微断裂现象;肝脏的肝细胞出现严重的水样变性,伴随着细胞溶解、浑浊变性和轮廓模糊,血窦扩张,形成局部的点状病灶;卵巢中卵母细胞出现核变形,胞质中出现小的空泡,细胞核膨胀,扩张和核仁空泡化;精巢中生精小管畸形,含不同发育阶段的精原细胞、精母细胞、精子细胞,精细胞体积增大,精子数量明显减少。随着Mo~(6+)质量浓度的升高和时间延长,草鱼肝胰脏中的超氧化物歧化酶活性和丙二醛含量降低。     

急性氨氮胁迫对虎斑乌贼肝脏、鳃和脑组织结构的影响

为了探讨氨氮对虎斑乌贼器官组织结构的影响,以体质量为(13.80±0.65)g的幼虎斑乌贼为对象,研究了氨氮胁迫对其肝脏、鳃和脑组织结构的影响。根据96 h的LC50实验结果设计5个梯度(0、1、3、6和12 mg/L)进行96 h的氨氮胁迫后,利用光学显微镜和透射电子显微镜观察其肝脏、鳃和脑组织结构。结果显示,在相同浓度的氨氮胁迫下,虎斑乌贼不同组织器官之间损伤程度存在差异,其中肝脏损伤的程度最大,对氨氮胁迫表现最敏感,其次是鳃组织和脑组织,组织器官损伤程度与氨氮浓度呈现正相关。氨氮胁迫浓度越大或胁迫时间越长,肉眼观察发现肝脏颜色越鲜红,肿胀和易糜烂程度越明显,可通过肉眼观察肝脏颜色和肿胀程度初步判断机体氨中毒程度;氨氮胁迫后,对肝脏和鳃组织造成较为严重的损伤,可能是虎斑乌贼氨中毒致死的原因。当氨氮高于或等于6 mg/L时胁迫96 h后,通过显微观察发现肝小叶轮廓模糊不完整、排列不紧密和胞浆疏松透明,大量的细胞核溶解,细胞出现空泡化,肝血窦扩张;通过电镜观察发现细胞核皱缩变形、核仁消失、线粒体嵴紊乱、线粒体空泡化、线粒体外室肿胀,高尔基体数量减少;显微观察发现鳃组织的泌氯细胞和上皮细胞核溶解,细胞出现空泡化、排列紊乱,鳃丝肿胀淤血、轮廓模糊不完整,并出现坏死脱落和缺损;通过透射电镜观察,发现细胞核皱缩、核膜破损和细胞核裂解,线粒体出现了皱缩变形、空泡化和不完整破损现象;观察脑组织的神经团和视叶,未发现脑组织的细胞有显著损伤。     

Pb2+对泥蚶鳃、肝脏等组织结构的影响

为了进一步了解重金属Pb对贝类的毒性毒理影响,研究了4个暴毒浓度(5、15、45和90μg/L)Pb2+对泥蚶鳃和肝脏组织显微结构的影响,及其中2个高浓度组Pb2+暴毒对泥蚶鳃和肝脏组织超微结构的影响,处理时间为96 h。结果显示,低浓度组Pb2+造成泥蚶鳃丝脱离其相连的软骨组织;高浓度组Pb2+使其鳃丝呼吸上皮细胞出现脱落,鳃腔肿胀、内有血细胞堆积,鳃丝断裂;超微结构分析发现,随着浓度的升高,鳃上皮细胞中次级溶酶体和线粒体数量均增加。Pb对泥蚶肝脏组织的显微结构影响仅见高浓度组肝小管附近有黄色物质沉积,消化管界限缺失,超微结构表明随着浓度的升高,肝细胞内沉积大量的黑色嗜锇性物质,次级溶酶体亦大量增加,细胞胞质及细胞核出现空泡,最后将导致细胞坏死,造成肝细胞永久性损伤。结合其他重金属毒性研究可发现,重金属引起的病理学变化并不具有金属特异性,主要还是氧化胁迫造成的氧化损伤。研究推测,生物体的抗氧化体系在重金属解毒中发挥了重要作用。     

久效磷对中国对虾细胞超微结构的影响：Ⅰ.对肝胰脏和肌肉的毒性效应

通过对０．１０ｍｇ／Ｌ久效磷处理的中国对虾细胞超微结构的透射电镜观察，结果表明：１中毒的肝胰脏细胞内质肉严重水肿、扩张、囊泡化，片层结构水肿；线粒体内嵴局部瓦解；肝管微绒毛出空缸区域及经物中毒的退化结构；脂滴电子密度不均。２中毒的肌肉细胞肌原纤维萎缩、间隔增大，肌浆网水肿、扩张、部分肌浆网膜破裂，线粒体内嵴膨胀，细胞质中出现中毒坏死巢。     

中华绒螯蟹肝胰腺白化症组织病理变化

采用常规石蜡切片技术和透射电镜技术,对患有肝胰腺白化症的中华绒螯蟹病变组织和细胞进行观察。发现病蟹肝胰腺、鳃、肌肉等组织细胞发生了不同程度的病变,其病理特征为肝细胞水肿、变性、坏死,细胞中脂肪颗粒锐减,线粒体水肿、嵴消失、形成空泡,内质网、溶酶体解体成小泡。鳃组织增厚,上皮细胞微绒毛排列疏松、不规则,部分线粒体嵴消失,出现膜性退化。坏死的肝细胞和鳃上皮细胞中出现细菌颗粒。肌细胞的病变主要表现为细胞核固缩,肌丝松驰、水肿、断裂、肌质网溶解形成小泡。中华绒螯蟹具有重要生理功能的肝胰腺、鳃组织发生了严重病变,使其功能损伤,不耐运输;而肌肉的松驰、肝胰腺脂肪的锐减影响了中华绒螯蟹的品质。同时,病变组织和细胞中未见病毒等生物病原,表明该病为非生物性疾病。图2表0参12     

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.