Histopathologic survey of ovaries of plaice, Pleuronectes platessa L., from Aber Wrac'h and Aber Benoit, Brittany, France: long-term effects of the Amoco Cadiz crude oil spill

Abstract. Ovarian samples were collected from plaice, Pleuronectes platessa , captured in the highly oiled Aber Wrac'h and Aber Benoit estuaries at four intervals during 1979–1980, following the Amoco Cadiz crude oil spill. Reference plaice were obtained along the western and southern coasts of Brittany. Tissue samples were fixed, processed and stained by routine histologic procedures. Ovaries were examined for histopathologic changes and for four different types of follicles. The concentrations and percentages of four types of ovarian follicles (primordial, primary, growing and mature) were determined for plaice ovaries within each sampling site at each sampling interval. Reference site ovaries, which were considered representative of the ovarian cyclic events for plaice along the Brittany coast, had primordial and primary follicles in high concentration in the summer season and in low concentration in the winter season. Concentrations of mature ovarian follicles in reference site plaice ovaries were low in the summer and high in the winter seasons. Growing follicles were observed infrequently in the reference site plaice ovaries. The seasonal pattern exhibited by follicles in ovaries from Aber Benoit plaice was similar to that observed in the reference site ovaries, but both the concentrations and percentages of follicle types were lower. The seasonal pattern of follicle types in Aber Wrac'h plaice ovaries was the reverse of that seen in the reference site plaice ovaries. In the Aber Wrac'h samples, concentrations of primordial and primary follicles were high in the winter months and low in the summer season, and no mature follicles were observed at any season. Growing follicles were seen more frequently in Aber Wrac'h and Aber Benoit plaice ovaries than in reference site plaice ovaries at all sampling intervals. Other than atretic follicles with an associated leucocytosis, no histopathologic conditions were observed in any ovary.     

Pathology of bacterial gill disease: sequential development of lesions during natural outbreaks of disease

Abstract. This study examines the sequential pattern of bacterial colonization and the sequential development of branchial lesions during typical outbreaks of bacterial gill disease (BGD) among four groups of commercially reared rainbow trout fingerlings, Oncorhynchus mykiss (Richardson). During a 5-month monitoring regime, prior to the onset of natural outbreaks of BGD, gill morphology of examined fish remained unaltered. Bacterial colonization of the gills was immediately preceded by several gill changes which were widely present in fish from all groups under study. These changes, which were detectable at the ultrastructural level only, included cytoplasmic blistering and degeneration of the microridges of the superficial filament epithelium, in addition to slight topographical irregularity of the filament tips suggestive of mild hyperplasia. Bacterial colonization began at these altered filament tips before spreading to more proximal regions of the filament and adjacent lamellar surfaces. Explosive increases in proportional morbidity and mortality coincided with the development of the following gill lesions: extensive bacterial colonization of lamellar surfaces, lamellar epithelial hydropic degeneration and necrosis, and lamellar oedema. In contrast, gill lesions such as lamellar fusion, epithelial hyperplasia and various metaplastic responses were detected later either as subacute (3–5 days) or chronic (7–14 days) changes. The role of epithelial necrosis as an important event during BGD is discussed as a possible mechanism leading to the development of sub-acute and chronic gill lesions.     

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。     

Branchial lesions associated with intermittent formalin bath treatment of Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum)

Two trials were conducted to assess the effects of repeated prophylactic formalin treatments on the gill structure of salmonids. In trial 1, which involved Atlantic salmon, Salmo salar L., approaching smoltification in a commercial facility, fish were treated with either 167 or 250 mg l^-1 formalin for 90 min every 2 weeks for 12 weeks. Formalin-treated salmon had slight, but not significant, increases in the frequency of lamellar fusion, numbers of lamellar mucous cells, and numbers of an endemic gill ciliate, Trichophyra piscium, after 6 and 12 weeks of treatment. In trial 2, which involved juvenile rainbow trout, Oncorhynchus mykiss (Walbaum), fish were treated with 200 mg l^-1 formalin for 60 min twice weekly for 12 weeks. Significant effects were limited to an increase in the numbers of mucous cells present on gill lamellae. In both trials, there was no evidence of lamellar oedema or necrosis of lamellar epithelial cells.     

Pathology of bacterial gill disease: ultrastructure of branchial lesions

Abstract. The range of branchial lesions associated with bacterial gill disease (BGD) in rainbow trout, Oncorhynchus mykiss (Richardson), was investigated through the ultrastructural examination of 23 separate outbreaks of the disease condition within commercially reared stocks. Diseased branchial tissues had a large and diffusely distributed monomorphic population of filamentous bacteria which were strictly epicellular. Although bacterial colonization was restricted to the branchial cavity, it was neither site nor cell specific: epithelial and chloride cells of the lamellae, filaments and lining tissues of the branchial cavity were all uniformly affected. The bacteria possessed an extensive glycocalyx which appeared to facilitate adhesion to the apices of the microridged sub-unit modifications of the cell surface. Sites of colonization were accompanied by a diffuse pattern of cellular degeneration and necrosis that was generally restricted to the outer layers of epithelium. The polarity and nature of these changes suggest that the mechanism of interaction between the bacteria and host cells includes progressive hydropic change as a sequel to primary membrane damage and consequent increased cell permeability. These cellular changes were accompanied by the range of stereotypical responses of the gill to damage frequently reported for BGD including lamellar fusion, epithelial hyperplasia, and squamous and mucous cell metaplasia, in addition to lamellar spongiosis.     

Development of a novel histopathological gill scoring protocol for assessment of gill health during a longitudinal study in marine-farmed Atlantic salmon (Salmo salar)

Gill health and disease in farmed Atlantic salmon (Salmo salar) were studied at a marine site on the west coast of Ireland. Fish were sampled on a regular basis during the summer and autumn of 2009. A novel histopathological scoring system was devised to give a semi-quantitative measure of gill pathology. The index criteria for gill histopathology were lamellar hyperplasia, lamellar fusion, circular anomalies (necrosis or sloughing) and lamellar oedema, a score from 0 to 3 being assigned for each parameter. Ancillary criteria, such as hypertrophy, haemorrhage and the presence of specific pathogens, were assigned a score of 0 or 1. Environmental monitoring undertaken included zooplankton and phytoplankton sampling, as well as temperature and oxygen recordings for the site. Epitheliocystis was frequently observed at low levels but was not associated with any significant gill pathology. Clinically significant gill pathology was observed on one occasion during the sampling period, coinciding with the occurrence of Pelagia noctiluca, Muggiaea atlantica and Solmaris corona. Throughout the longitudinal study, the scoring system provided an accurate and effective means of assessing gill health, demonstrating the ability of this practical tool to be used in future studies of gill health and disease.     

Morphometric and histochemical assessment of the branchial tissue response of rainbow trout, Oncorhynchus mykiss (Walbaum), associated with chloramine-T treatment

Twelve tanks of juvenile rainbow trout, Oncorhynchus mykiss (Walbaum), were treated twice weekly for an 11- week period with a one-hour static bath of chloramine-T (10 mg l^–1). The morphometric indices of gill damage and the histochemical characteristics of gill mucous cells of fish from treated and paired untreated tanks were compared. The use pattern of chloramine-T in this study evoked a slight increase in mean lamellar width, but it did not induce a greater degree of lamellar oedema, lamellar fusion, tissue infiltration, epithelial hyperplasia, chloride cell metaplasia or thrombosis of pillar channels in treated fish. Treatment caused a trend towards an increased number of mucous cells on lamellae, associated with a significant shift from neutral mucin to acid mucin production based on histochemical characteristics.     

氯氰菊酯暴露对草鱼4种器官组织结构的影响

通过氯氰菊酯对草鱼(Ctenopharyngodon idellus)幼鱼(9.30±0.48)g的急性毒性实验及对鳃、肝、肾、脾的组织学研究,探讨了随着氯氰菊酯浓度的增加以及暴露时间的延长草鱼组织结构损伤的变化趋势.结果显示:氯氰菊酯对草鱼种的48 h-LC50和96 h-LC50分别为55.21μg/L和25.00...     

Pathology of experimental amoebic gill disease in Atlantic salmon, Salmo salar L., and the effect of pre-maintenance of fish in sea water on the infection

Atlantic salmon were exposed to amoebic gill disease (AGD) immediately following their acclimatization to sea water (group 1), or following a 2 week period of maintenance in sea water (group 2). Three fish from each group were sampled on days 0, 1, 2, 4, 7, 14 and 28 post-infection. Characteristic gill lesions began to occur between days 2 and 4, and dramatically increased by day 7. The number of gill lesions on fish from group 2 was significantly higher than on fish from group 1 on days 7 and 14 ( P <0.001), but the two groups did not differ in any other parameter. Histologically, Paramoeba sp., the aetiological agent of AGD, could be seen on the gills of fish as soon as 1 day post-exposure, attached to healthy-appearing gills. Gill pathology in the form of hyperplasia and lamellar fusion followed shortly. AGD infection was accompanied by a significant increase in the number of gill mucous cells ( P =0.002). Different methods for the diagnosis of AGD are discussed.     

Histopathological effects of cadmium on the gills of the freshwater fish, Macropsobrycon uruguayanae Eigenmann (Pisces, Atherinidae)

Evaluations of histopathological lesions in gill tissue were carried out in the freshwater fish Macropsobrycon uruguayanae following 30 and 60 days of exposure to 1.5 mg 1^-1 of cadmium. The study was conducted on both fed and starved animals in order to determine the influence of feeding condition on cadmium toxicity. The main lesions observed and quantified were: (1) hyperplasia of primary lamellar epithelium; (2) hyperplasia of secondary lamellar epithelium; (3) separation of respiratory epithelium; (4) shortening of secondary lamellae; (5) epithelial necrosis; (6) fusion of adjacent secondary lamellae; (7) hypertrophy of respiratory epithelium; (8) lamellar telangiectasis; (9) hyperplasia of chloride cells; (10) mucinous metaplasia; and (11) inflammatory infiltration. Lesions 6, 8, 9 and 11 were only induced by exposure to cadmium, while lesion 4 could be produced only by starvation. Starved fish also showed a reduction in total body weight and length. Lesion 2 was shown to be non-specific, and produced by either cadmium, starvation or even exposure time. The possible mode of action of the experimental factors are discussed in relation to the observed pathologies.     

Acid water and aluminium exposure: gill lesions and aluminium accumulation in farmed brown trout, Salmo trutta L.

Abstract. Brown trout, Salmo trittta L., from two fish farms, situated within acid-susceptible areas in Sweden, were studied with respect to aluminium accumulation and gill pathology. The total aluminium concentrations, after liming of the water in both farms, were between 200 and 300μg/1 at sampling. Brown trout from a third farm in a non-acidified area (total aluminium concentration of water 35μg/1 were included as a control. The aluminium concentrations in gills from farmed brown trout in acidified areas were approximately 20μg/g and 90μg/g wet weight respectively compared with approximately 2μg/g wet weight in control brown trout gills. Light- and electron microscopic examination disclosed two major types of gill lesions characterized by chloride cell hyperplasia in the secondary lamellar epithelium and enlargement of the intercellular spaces in the secondary lamellar epithelium. The difference in the aluminium speciation and accumulation may explain the difference in the gill pathology of farmed fish from acidified areas. The chloride cell hyperplasia together with a relatively low aluminium accumulation is considered to reflect a long-term exposure effect.     

鲫圆形碘泡虫病组织病理学研究

采用常规石蜡切片、苏木精-伊红染色的方法,研究了鲫（ Carassius auratus auratus ）自然感染圆形碘泡虫（Myxobolus rotundus）后的组织病理变化。结果表明,感染圆形碘泡虫后,在鲫的头部、鳃和体表能形成大小不一的圆形或椭圆形乳白色孢囊,其孢囊为典型的3层结构：外层的结缔组织、双层质膜的外质以及各个发育阶段的孢子或营养体组成的内质。病理切片可见鱼体各组织器官均有不同程度的病变,主要表现为鳃丝粘连、肿胀增生;肝脏发生以肝血窦为中心的透明、空泡变性;肾小管内皮脱落,肾小球萎缩;肠道粘膜层脱落,肌层细胞肿胀病变;脾脏的含铁血黄素增多等异常现象。     

Effect of buffers on the gill structure of common carp, Cyprinus carpio L., and rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract. The effects of phosphate and tris buffers on the gill structure of common carp, Cyprinus carpio L, and rainbow trout, Oncorhynchus mykiss (Walbaum), were investigated. Common carp maintained in phosphate-buffered (a combination of sodium di-hydrogen phosphate and di-sodium hydrogen phosphate at a final concentration of 0.01 m ) standard dilution water for a period of 8 days developed severe pathological changes in the gill tissues, while carp maintained in tris-buffered (0.01 m ) dilution water and trout subjected to either of these buffers showed no such changes. Histological changes in the gills of carp exposed to phosphate buffered dilution water included an extensive mucous cell proliferation on the lamellar and filarnental epithelium, and an increased infiltration of eosinophilic granular cells into the gill tissues. A moderate epithelial hyperplasia and an increase in the chloride cell densities on the gill epithelia were also noted. The implications of these findings on the widely accepted practice of the use of phosphate salts and various other buffers in pH control of experimental water are highlighted.     

拟态弧菌感染黄颡鱼的动态病理损伤及病原分布研究

以1.0×106CFU·m L~(-1)拟态弧菌(Vibrio mimicus)浸浴感染黄颡鱼(Pelteobagrus fulvidraco),于第0、第4、第8、第16、第24、第36、第48、第60、第72小时剖杀取样,研究病理损伤及病原菌的体内动态分布。结果显示,皮肤肌肉、鳃、肠道出现病变最早,其中皮肤肌肉损伤最严重。第16小时表皮变性、坏死,鳃上皮肿胀,肠绒毛水肿;第24~第48小时表皮坏死、脱落,真皮严重出血,肌纤维变性、坏死,鳃出血,上皮局部坏死,肠上皮变性与灶性坏死;第48~第72小时皮肤肌肉坏死更严重,形成溃疡灶,鳃灶性坏死。肾、肝、脾与心36 h后相继出现不同程度的淤血、出血,变性和灶性坏死。q PCR检测发现,第4小时即在鳃、肠、皮肤肌肉检出病菌,细菌含量分别为1.3×102CFU·mg~(-1)、2.6×102CFU·mg~(-1)、4.7×102CFU·mg~(-1);鳃与皮肤肌肉中菌量随时间延长而增加,第72小时皮肤肌肉中菌量达到4.5×107CFU·mg~(-1);第24小时后相继在肾、肝、脾、心检出病菌,菌量介于1.9×10~4.7×102CFU·mg~(-1)之间。结果表明皮肤、鳃和肠道是病菌的入侵位点,并在体内多组织、器官分布。     

Evaluation of dietary histidine supplementation on meagre,Argyrosomus regius,juvenile growth,haematological profile,stress and muscle cellularity

Histidine plays an important role in haemoglobin synthesis, antioxidant defence system and stress resistance, and it is thought to be a limiting amino acid in most meagre diets. In this study, two different diets, a control and a diet supplemented with 3% histidine, were tested in 60‐day‐old meagre (initial weight—6.2 g) for 82 days. Survival varied between 92% and 94%, and neither SGR nor whole fish composition seemed to be affected by the diet. When fish were submitted to a stress test (outside water for 30 s), it was observed that the levels of glucose and lactate significantly increased in both treatments. The concentration of cortisol increased after the stress test only in the His 30 g/kg group. Muscle cellularity was affected by the diets, where the control group showed a higher mean fibre area and lower fibre density. These results, associated with a lower frequency of smaller fibres, suggest a decrease in fibre recruitment and a higher fibre hypertrophy. In conclusion, the supplementation of diets with 3% led to a more pronounced response to the stress test in terms of cortisol levels and increased muscle fibre recruitment, decreasing muscle fibre hypertrophy contribution to muscle growth.     

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。     

Epitheliocystis disease in the cultured amberjack, Seriola dumerili Risso (Carangidae)

Epitheliocystis infection affecting the gill and the pseudobranch of the cultured amberjack, Seriola dumerili Risso, is described. In hyperinfected fish, proliferative cell response around the epitheliocystis capsule resulted in gill and pseudobranch lamellar fusion, which led to mass mortalities in the 0+age class. Histopathological and scanning electron microscope observations showed that the target cell was the chloride cell since (1) epitheliocystis organisms were first found within the chloride cell; (2) chloride cells underwent degeneration in the filament epithelium, proliferated along the lamellae, and hypertrophied; and (3) cysts were found only in the trailing edge of the gill filament and in the interlamellar spaces where, in healthy fish, chloride cells are mainly located. In some cases, the pseudobranch of infected fish was much more severely affected than the gill. It is therefore suggested that, for diagnostic purposes, the pseudobranch should also be sampled when epitheliocystis disease is suspected.     

Variations in mucous cell numbers in gills of Atlantic salmon (Salmo salar) presmolt in commercial freshwater farms in Norway

Fish gills are heavily exposed to the external milieu and may react against irritants with different cellular responses. We describe variations in mucous cell counts in gills from healthy Atlantic salmon (Salmo salar) presmolts in five recirculating aquaculture system (RAS) farms and one flow‐through farm. Based on certain criteria, mucous cells were histologically quantified in a defined lamellar region of the gills and the counts were analysed. Immunohistochemistry (IHC) was used to investigate epithelial responses. The median number of total mucous cells in the defined region was 59 per fish. Between the farms, the medians varied from 31 to 101 with the lowest in the flow‐through farm. A regression model was fitted with “total mucous cells” as the dependent variable and with “fish length” and “fish farm” as independent variables. The proportion of variation in mucous cell counts explained by the model was twice as high when “fish farm” was included compared to only “fish length.” IHC revealed proliferative responses in coherence with high mucous cell numbers. Conclusively, the variation in mucous cell counts depends on combined farm‐related factors. Establishing a baseline for mucous cell counts is fundamental in the development of high‐throughput monitoring programmes of gill health in farmed fish.     

硫酸铜蓄积对日本[虫寻]4种组织细胞超微结构的影响观察

通过蓄积毒性试验,研究了安全质量浓度（0．5mg·L^-1）下硫酸铜（CuSO4·5H2O）对日本[虫寻]（Charybdis japonica）4种组织细胞（肌肉、鳃、肝胰脏和心脏）超微结构的影响。结果表明,铜离子（Cu^2＋）蓄积主要损伤的组织是鳃和肝胰脏,其次是心脏和肌肉。鳃细胞的损害表现为鳃丝水肿,细胞器溶解,角质层变薄,线粒体、内质网的肿胀、解体;肝胰脏细胞的主要损害特征为肝管微绒毛减少,线粒体水肿解体,内质网扩张,细胞核空泡化,核膜水肿和脂肪滴增加;心脏细胞的毒理变化为线粒体内嵴肿胀、瓦解,肌原纤维不规则,内质网溶解。0．5mg·L^-1的CuSO4·5H2O经过16d蓄积后虽然没有导致日本[虫寻]死亡,但已经对其体内组织细胞的超微结构产生了不同程度的影响。