异源四倍体银鲫外周血和精巢的组织学特征

通过比较D 系三倍体银鲫 (Carassius auratus gibelio Bloch) 与异源四倍体银鲫, 我们发现异源四倍体的外周血与精巢组织跟三倍体银鲫存在明显差异。HE 染色结果表明, 异源四倍体银鲫外周血红细胞有明显的分裂倾向。利用流式细胞术对D 系三倍体银鲫与异源四倍体银鲫外周血的DNA 直方图进行比较, 结果表明异源四倍体外周血的DNA 直方图有两个主峰。此外, 我们观察到异源四倍体银鲫精巢的三种类型, 其中Ⅰ型精巢可以产生正常精子, Ⅱ型可观察到精小囊结构, 但不能产生精子, Ⅲ型精巢未发育出精小囊结构。进一步用银鲫Vasa 抗体对精巢切片进行组织免疫荧光共聚焦显微分析, 结果表明, Ⅰ型精巢的生殖细胞完成了减数分裂, 能观察到精原细胞、初级精母细胞、次级精母细胞, 以及大量位于精小管中间的精子细胞和精子; 而Ⅱ型精巢的生殖细胞不能完成第二次减数分裂, 精小囊中存在大量的初级和次级精母细胞, 没有精子细胞产生。研究丰富了对异源四倍体银鲫生物学性状的认识。       

银鲫精母细胞型精原细胞瘤的病理学观察

自银鲫(Carassius gibelio)中发现了一例自发性精母细胞型精原细胞瘤, 详细描述了该肿瘤的组织病理和超微结构。在组织学上, 该肿瘤具有薄膜包被、扩张性生长, 具有高度细胞性和弥漫性结缔组织, 伴有淋巴细胞浸润。透射电镜显示, 该肿瘤显著的特征是含有未分化的精母细胞, 且支撑细胞稀少。对比正常银鲫的精巢组织结构, 排除了其他肿瘤类型, 最终将该肿瘤鉴定为精母细胞型精原细胞瘤。系在低等脊椎动物中首次发现具有淋巴细胞浸润的精母细胞型精原细胞瘤, 丰富了鱼类性腺肿瘤的基础资料, 也为构建人类精母细胞型精原细胞瘤动物模型提供了依据。     

The development of carp gonads in warm water aquaria

The development of carp gonads in warm water (23°C) aquaria under controlled conditions is described. In this environment males mature at 6 months of age. Females ripen at 14 months, and by 15 months 25% of these have started to spawn, whereas under natural conditions female European carp do not ripen until they are 4 years old. Fishes do not lay all their eggs at once, as evinced by the finding of partly-spent ovaries. Sexual maturation of the ovaries has been described in six stages, based on histological changes, and associated with changes in the female gonado-somatic index and the maximum ova diameter. The maturation of testes occurs in four stages. The percentage incidence of each stage of ovarian maturation has been calculated for the 6th to the 15th month. Larger fishes become sexually mature before smaller fishes.     

Vascularization and related distribution of leucocytes in carp, Cyprinus carpio L., head kidney

The distribution of lymphoid cells in the carp head kidney was investigated in relation to the vascular system. Blood vessels in the head kidney were histologically identified into arteries, sinusoids and two types of veins: renal veins and portal, which were distinguished by India ink injection into the caudal vein. By histological and histoplanimetrical observations it was found that the head kidney contained a number of lymphoid cells, which mainly aggregate around the connections between the portal veins and sinusoids, and that the cellular density of the aggregations was higher than in the thymus.
Pigment-containing cell clusters were also observed around these connections. This arrangement of the blood vessels suggests that it is one of the structures able to trap foreign materials, and the occurrence of the lymphoid clusters around the portal veins is a phylogenetic sign of the morphological division between granulopoietic and lymphatic tissues in the carp head kidney.     

Histopathology of Ichthyophthirim multifiliis infections in fishes

Histopathological changes involved in Ichthyophthirius multifiliis infections are described from a wide range of host fishes—carp, crucian carp hydrids, Carassius carassius × C. auralus , goldfish, eels, African catfish, Glorias lazera , several species of cichlid fishes and rainbow trout. Fish were obtained from diverse geographical regions: Israel, South Africa, Portugal and Venezuela, as well as from laboratory experimental infections. Observed histopathological changes could not demonstrate any interspecific differences, but individual fish or different groups of fish of the same species demonstrated wide variability in susceptibility to infection and consequently in the nature and severity of the histopathological response. Low to moderate primary infection and secondary reinfestations did not induce significant histopathological changes in the integument. The proliferative response of the integumentary epithelium appeared only following subsequent infections. Repeated heavy reinfections induced massive cellular necrosis and in very heavy infections extensive lysis of the epithelial tissue. Host-parasite interactions in the light of the histological findings are discussed.     

Functional morphology of durophagy in black carp,Mylopharyngodon piceus

The black carp, Mylopharyngodon piceus (Osteichthyes: Cyprinidae), crushes its snail and other molluscan prey with robust pharyngeal jaws and strong bite forces. Using gross morphology, histological sectioning, and X‐ray reconstruction of moving morphology (XROMM), we investigated structural, behavioral, and mechanical aspects of pharyngeal jaw function in black carp. Strut‐like trabeculae in their pharyngeal jaws support large, molariform teeth. The teeth occlude with a hypertrophied basioccipital process that is also reinforced with stout trabeculae. A keratinous chewing pad is firmly connected to the basioccipital process by a series of small bony projections from the base of the pedestal. The pharyngeal jaws have no bony articulations with the skull, and their position is controlled by five paired muscles and one unpaired median muscle. Black carp can crush large molluscs, so we used XROMM to compare pharyngeal jaw postures as fish crushed ceramic tubes of increasing sizes. We found that black carp increase pharyngeal jaw gape primarily by ventral translation of the jaws, with ventral rotation and lateral flaring of the jaws also increasing the space available to accommodate large prey items. A stout, robust ligament connects left and right jaws together firmly, but allows some rotation of the jaws relative to each other. Contrasting with the pharyngeal jaw mechanism of durophagous perciforms with fused left and right lower pharyngeal jaws, we hypothesize that this ligamentous connection may serve to decouple tensile and compressive forces, with the tensile forces borne by the ligament and the compressive forces transferred to the prey. J. Morphol. 276:1422–1432, 2015. © 2015 Wiley Periodicals, Inc.     

Bone resorption and bone remodelling in juvenile carp, Cyprinus carpio L.

The present study considers the important role of bone resorption for bone growth in general, and aims to clarify if and how bone resorption contributes to the skeletal development of carp, Cyprinus carpio L., a teleost species with ‘normal’ osteocyte‐containing (cellular) bone. To ensure the identification of osteoclasts and sites of bone resorption independently from the morphology of the bony cells, bones were studied by histological procedures, and by demonstration of the enzymes which serve as osteoclast markers, viz. tartrate resistant acid phosphatase (TRAP), ATPase and a vacuolar proton pump. Two types of bone‐resorbing cells were observed in juvenile carp: (1) multinucleated giant cells displaying morphological and biochemical attributes which are known from mammalian osteoclasts; and (b) flat cells which lack a visible ruffled border and for which identification requires the performance of enzyme histochemical procedures. Bone resorption performed by osteoclasts mainly occurs at endosteal bone surfaces. To a lesser extent, bone resorption also takes place at periosteal bone surfaces, but without an apparent connection to bone growth. The latter observation, and the occurrence of bone remodelling, suggest that the endoskeleton of juvenile carp might be involved in mineral metabolism. Morphological differences and biochemical similarities to bone resorption in teleosts with acellular bone are discussed.     

Comparative studies on histological and ultra-structure of the pituitary of different ploidy level fishes

The histological and ultra-structure of the pituitary in diploid red crucian carp(Carassius auratus red var.),triploid crucian carp and allotetraploid hybrids within and after the breeding season were comparatively studied.The result showed that there were six endocrine cell types in the pituitary of these three kinds of fishes,and there was an obvious difference in cell size among different ploidy level fishes.As for the same type of pituitary cells,the cell size was increased gradually with the in- creasing ploidy level.In the breeding season,the allotetraploid hybrids had higher proportion of go- nadotropin cells(GTH)than triploids,and the triploids had higher proportion of GTH than diploids.The results were related to the earlier sexual maturity of allotetraploid hybrids and sterility of triploid cru- cian carp.On the other hand,among the three kinds of fishes,the proportion of somatotropin(STH) cells in triploids crucian carp was the highest,whereas that in allotetraploid hybrids was the lowest. The results might be connected with the faster growth rate of triploids and slower growth rate of al- lotetraploid hybrids.In addition,in GTH cells of meso-adenohypophysis after the breeding season, there were many endocrine particles in triploids,while those endocrine particles were released from the cells in allotetraploids and diploids.This result showed that the sterility of triploid crucian carp might be related to the hormone which was not released from the GTH cells.In a word,the present study indicated that the differences in the structure of pituitary among different ploidy level fishes contributed to their difference in the growth rate and gonadal development.     

Comparative studies on histological and ultra-structure of the pituitary of different ploidy level fishes

The histological and ultra-structure of the pituitary in diploid red crucian carp (Carassius auratus red var.), triploid crucian carp and allotetraploid hybrids within and after the breeding season were comparatively studied. The result showed that there were six endocrine cell types in the pituitary of these three kinds of fishes, and there was an obvious difference in cell size among different ploidy level fishes. As for the same type of pituitary cells, the cell size was increased gradually with the increasing ploidy level. In the breeding season, the allotetraploid hybrids had higher proportion of gonadotropin cells (GTH) than triploids, and the trploids had higher proportion of GTH than diploids. The results were related to the earlier sexual maturity of allotetraploid hybrids and sterility of triploid crucian carp. On the other hand, among the three kinds of fishes, the proportion of somatotropin (STH) cells in triploids crucian carp was the highest, whereas that in allotetraploid hybrids was the lowest. The results might be connected with the faster growth rate of triploids and slower growth rate of allotetraploid hybrids. In addition, in GTH cells of meso-adenohypophysis after the breeding season, there were many endocrine particles in triploids, while those endocrine particles were released from the cells in allotetraploids and diploids. This result showed that the sterility of triploid crucian carp might be related to the hormone which was not released from the GTH cells. In a word, the present study indicated that the differences in the structure of pituitary among different ploidy level fishes contributed to their difference in the growth rate and gonadal development.     

Morphological,histological and molecular characterization of three Myxobolus species (Cnidaria: Myxosporea) from silver carp Hypophthalmichthys molitrix Valenciennes and bighead carp Hypophthalmichthys nobilis Richardson in China

Three Myxobolus species were obtained from silver carp Hypophthalmichthys molitrix Valenciennes and bighead carp Hypophthalmichthys nobilis Richardson in China. In the present study, we supplemented their taxonomic characteristics by the morphological, histological and molecular methods. Myxobolus kiuchowensis Chen in Chen et Ma, 1998 formed small ellipsoidal plasmodia in the intestinal wall of bighead carp. Its spores appeared asymmetrical obovate in frontal view and fusiform in lateral view. Tiny mamillary protrusion in the anterior of some spores was observed. Two pyriform polar capsules were unequal. Histologically, M. kiuchowensis infected the tunica muscularis of host intestine. Myxobolus abitus Li et Nie, 1973 formed sausage–like plasmodia in the gills of silver carp. Its spores appeared oblate in frontal view and fusiform in lateral view. Two pyriform polar capsules were unequal and an obvious inter–capsule appendix was observed. Histological examination revealed that M. abitus developed in the interlamellar–epithelium of host gills. Myxobolus pavlovskii (Akhmerov, 1954) Landsberg et Lom, 1991 formed sausage–like plasmodia both in the gills of silver carp and bighead carp. Spores of M. pavlovskii were proximate oval in frontal view and fusiform in lateral view. Two pyriform polar capsules were unequal. The BLAST search indicated the SSU rDNA sequences of M. kiuchowensis and M. abitus were not identical to any sequence, however, the SSU rDNA sequences of M. pavlovskii were identical to that of M. pavlovskii recorded previously. Phylogenetic analysis showed that the present three species robustly clustered together in Cyprinid group and Asia group.     

New type of pathogenicity of Thelohanellus kitauei Egusa & Nakajima, 1981 infecting the skin of common carp Cyprinus carpio L.

Thelohanellus kitauei Egusa & Nakajima, 1981 is a common parasite infecting the intestine of common carp Cyprinus carpio L., resulting in mass mortality or loss of economic value of cultured carp. In the present study, T. kitauei infecting host skin was detected. The morphological, molecular and histological data of this parasite in the new organ record are presented. Morphological analysis showed the current specimen morphologically similar to T. kitauei from the intestine. Despite the spore length and polar capsule length of the current specimen larger than those of T. kitauei from the intestine, ranges of dimensions overlap, which is more suggestive of intraspecific variation than distinct species. BLAST search revealed that the present small subunit ribosomal DNA gene sequence is identical to those of T. kitauei. Histologically, most of spores distributed in the stratum spongiosum of dermis, and some spores in the strata compactum of host skin were also observed. Above all, both morphology and molecular analysis indicated that the current species from the skin of common carp is conspecific with T. kitauei from the intestine of carp and organ habitats transfer of T. kitauei from host intestine to skin may have occurred.     

鲤鱼卵黄发生期血钙水平调节初步研究

在雌鲤卵黄发生期的10月～次年1月和斯坦氏小体(CS)发育静止期进行升高细胞外钙浓度实验,分别用OCPC法测定血浆钙水平的变化,超微结构观察CS的组织学变化.结果表明,鲤鱼血钙水平在卵黄发生期存在一定性别差异,雄鲤水平普遍低于雌鲤,在11月达显著性水平.升高细胞外钙浓度的各处理组中CS的超微结构发生了不同程度的变化,均表现为细胞代谢活动增强.     

尼罗罗非鱼性腺发育的研究

湖南地区生长于池塘环境的尼罗罗非鱼(Tilapia nilotica),性成熟日龄是110—130天,雄性比雌性普遍早熟20天。精细胞的发生能够完成由精原细胞到精子的全部发育过程,同样,卵细胞的发生也能完成由卵原细胞到卵子的全部发育过程。初级卵母细胞处于Ⅲ时相阶段时,可由包被卵周的滤泡细胞分泌产生放射膜,但放射膜不在动物性极形成受精孔,也无精孔细胞的分化,证实尼罗罗非鱼是属于非受精孔受精类型。初级卵母细胞由Ⅲ时相发育到Ⅳ时相是非同步性的,产后卵巢的组织学结构为第Ⅳ期,卵巢系数在繁殖季节可出现三次高峰,证实尼罗罗非鱼是属于多次产卵类型。通过对精巢组织学的研究,发现尼罗罗非鱼的第Ⅰ期精巢是自然两性嵌合体。       

Sublethal effects of endosulfan on the histology and protein pattern of Cyprinus carpio gill

Histological and protein pattern changes in the gill of the common carp, Cyprinus carpio were investigated following exposure to sublethal concentrations of the organochlorine insecticide, endosulfan. Hyperplasia, lamellar fusion, curling of the tips of primary lamellae, exudation of erythrocytes and bulging of the tips of primary lamellae were the histological lesions observed. Protein pattern of the gill varied at different sampling periods characterised by disappearance of certain fractions and occurrence of additional fractions. It was concluded that the severity of histological changes and protein pattern variations seen in the gill was dependent on both the duration of exposure and the concentrations of endosulfan used (over the range 5—25 % of the 96 hr LC₅₀ value, 5·199ppb).     

急性氨氮胁迫对于草鱼sod和hsp90基因表达及鳃部结构的影响

实验通过测定草鱼的24h半致死浓度, 鳃的细胞结构以及sod和hsp90的表达模式研究了草鱼在组织学和分子生物学水平上对高浓度氨氮暴露的响应。经过半致死实验确定的氨氮24h LC50为243 mg/L试验中草鱼被置于5个浓度的处理组中(50、72、104、151、220 mg/L), 之后取鳃组织进行组织切片分析, 取肝脏、肠和鳃来测定sod和hsp90的表达情况。经过高浓度的氨氮暴露处理, 鳃组织的细胞排列和结构产生了明显的变化, 并且sod和hsp90的表达受到了显著的上调。这些结果表明, 高浓度的氨氮能够损害鳃部的细胞结构并且诱导应激蛋白的表达。这个结果同样显示出, sod和hsp90可以作为评估草鱼氨氮暴露水平的良好指标。       

A histological and morphometric study on the tissue and cellular distribution of iron in carp Cyprinus carpio L. during chronic waterborne exposure

The effect of long-term (77 day) exposure of carp Cyprinus carpio to low concentrations of waterborne iron (1 mg Fe-dextran l⁻¹) on accumulation and cellular distribution of iron in the liver was assessed using Perl's staining and histological observations and morphometric measurements. Accumulation of iron in the liver occurred after 14 days of exposure, when the majority of the iron was found in the sinusoidal endothelium with lower amounts in the cytoplasm and nuclei of hepatocytes. Upon further treatment the iron was predominantly distributed in the cytoplasm of the hepatocytes either as granules or diffusive iron and in macrophages.     

饲喂不同浓度黄曲霉毒素B1饲料对异育银鲫成鱼的生长和毒素积累的影响

以含不同浓度黄曲霉毒素B1(AFB1)的配合饲料饲喂异育银鲫(Carassius auratus gibelio)成鱼56d,研究异育银鲫成鱼[(122.3±0.7)g]生长、生理反应、肝脏组织学变化、卵巢发育以及鱼体各组织中的AFB1的毒素积累状况。实验分为5个实验组,不同实验组饲料中AFB1含量分别为0、5、20、50、500μg/kg饲料(实测值分别为2.59、4.12、12.39、46.23、454.07μg/kg饲料),每个处理3个平行。在整个实验过程中各实验组均未表现出外部形态和行为异常,各组存活率均达到100%。各实验组异育银鲫成鱼终末体重、摄食率(FR)、特定生长率(SGR)和饲料效率(FE)均无显著差异。饲料AFB1水平对异育银鲫血清总胆固醇(TC)含量、血清谷丙转氨酶(GPT)、谷草转氨酶(GOT)和碱性磷酸酶(AKP)活性均无显著影响。各毒素组血清超氧化物岐化酶(SOD)活性与对照无显著差异。各毒素组肝脏和卵巢均未见明显的组织学病理变化。肌肉和性腺中的AFB1积累量低于FDA食品安全限定标准(5μg/kg)。肝胰脏中的AFB1积累和饲料中的AFB1水平呈对数关系。饲喂AFB1≥50μg/kg饲料使异育银鲫成鱼肝脏AFB1积累超过安全限量标准。结果表明,异育银鲫成鱼至少可耐受AFB1含量达500μg/kg饲料(实测值:454.07μg/kg饲料)56d。     

Identification of keratins and analysis of their expression in carp and goldfish: comparison with the zebrafish and trout keratin catalog

With more than 50 genes in human, keratins make up a large gene family, but the evolutionary pressure leading to their diversity remains largely unclear. Nevertheless, this diversity offers a means to examine the evolutionary relationships among organisms that express keratins. Here, we report the analysis of keratins expressed in two cyprinid fishes, goldfish and carp, by two-dimensional polyacrylamide gel electrophoresis, complementary keratin blot binding assay, and immunoblotting. We further explore the expression of keratins by immunofluorescence microscopy. Comparison is made with the keratin expression and catalogs of zebrafish and rainbow trout. The keratins among these fishes exhibit a similar range of molecular weights and isoelectric points, with a similar overall pattern on two-dimensional gels. In addition, immunofluorescence microscopy studies of goldfish and carp tissues have revealed the expression of keratins in both epithelial and mesenchymally derived tissues, as reported previously for zebrafish and trout. We conclude that keratin expression is qualitatively similar among these fishes, with goldfish and carp patterns being more similar to each other than to zebrafish, and the cyprinid fishes being more similar to each other than to the salmonid trout. Because of the detected similarity of keratin expression among the cyprinid fishes, we propose that, for certain experiments, they are interchangeable. Although the zebrafish distinguishes itself as being a developmental and genetic/genomic model organism, we have found that the goldfish, in particular, is a more suitable model for both biochemical and histological studies of the cytoskeleton, especially since goldfish cytoskeletal preparations seem to be more resistant to degradation than those from carp or zebrafish. This work was supported by grants to J.M. from the Stiftung Rheinland Pfalz für Innovation (836-386261/138) and the Deutsche Forschungsgemeinschaft (Ma 843/5-1) and a grant to D.G. from the National Science Foundation (INT-0078261).     

饲喂不同浓度黄曲霉毒素B1饲料对草鱼幼鱼生长和毒素积累的影响

以含不同浓度黄曲霉毒素B₁(AFB₁)(0、10、20、100、1000和5000 μg/kg饲料)的6种等氮等能(32.96%蛋白质, 14.55 kJ/g能量)配合饲料饲喂平均初始体质量为(2.90±0.16) g草鱼(Ctenopharyngodon idellus)幼鱼84d, 探讨AFB₁对草鱼幼鱼生长、肝胰脏和肾脏组织结构以及鱼体肌肉中的毒素积累的影响。实验分为6个实验组, 每组3个平行。结果表明, 在整个实验过程中各实验组幼鱼的行为均未表现出异常, 各组幼鱼的存活率、终末体重、摄食率、特定生长率、饲料效率、肝体比、脏体比均无显著差异。饲料AFB₁水平对草鱼幼鱼血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(AKP)、超氧化物岐化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性均无显著影响。各毒素组和对照组肝胰脏、肾脏组织学观察中未发现病理变化。摄食AFB₁≤1000 μg/kg的草鱼幼鱼肌肉中未检测出AFB₁残留, 仅在5000 μg/kg实验组中检测出肌肉中含有(1.21±0.18) μg/kg的AFB₁, 低于FDA食品安全限定标准。由此可见, 草鱼幼鱼至少可耐受AFB₁含量达5000 μg/kg饲料(实测值: 4979.2 μg/kg饲料) 84d。