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1.
对自然发病、投喂感染和注射感染的中国对虾、凡纳滨对虾白斑综合征进行组织与细胞病理研究，结果发现：中国对虾与凡纳滨对虾的病理变化相似，但不同感染方式的患病对虾有差异。临床病理：相同点表现为游动无力，反应迟钝，胃中无食，体色变暗，肌肉浑浊，肝胰腺肿大；不同点为自然发病和投喂感染的对虾临床病理变化比注射人工感染明显，病程长。显微病理：相同点表现为细胞核肿大，细胞变性、坏死等；不同点为自然发病和投喂感染的病虾胃、肝胰腺的病理变化比注射感染严重，坏死细胞数量多，组织大面积坏死溶解；注射感染病虾的肌肉组织显微病理变化比自然发病、投喂感染严重。超微病理：相同点表现为细胞肿大、变性、坏死、溶解，线粒体、内质网等细胞器形态变异，肿大，膜溶解或整体崩解；细胞核肿大或固缩或溶解．部分细胞核中可见WSSV；不同点为注射感染的患病对虾肌肉组织的超微病理变化比自然发病、投喂感染严重，肌细胞核中观察到WSSV的几率较高；投喂感染、自然发病对虾的胃上皮细胞、肝胰腺细胞的超微病理变化比注射感染严重。另外，不同类型的细胞对WSSV的易感性不相同，血细胞为最常见的被病毒感染的靶细胞，血细胞通过细胞变形或形成伪足，贴附于其它组织细胞膜上。 相似文献
2.
草鱼吻端组织细胞株ZC-7901及其亚株ZC-7901S1的建立和特性观察 总被引：5，自引：0，他引：5
以草鱼吻端组织为材料，经体外培养建立了一个上皮样细胞和梭形细胞混合的细胞株，后又从中分离出一个上皮样细胞亚株，分别定名为ZC-7901和ZC-7901S1。对两株细胞进行了基本生物学特性的测定，染色体数目和经长期传代后染色体数目稳定性的分析。实验表明，两株细胞为草鱼正常二倍体(2n=48)细胞株。 相似文献
3.
对对虾病毒性疾病的研究目前国内尚属空白，国外也只在七十年代后期才出现为数不多的报道。初步统计主要有对虾杆状病毒、斑节对虾杆状病毒和传染性皮下及造血器官坏死病毒等。这几种病毒形态特征基本相似，都属于杆状病毒，只是寄主的种类及组织病理学上稍有不同。这些病毒的致病特点都是侵袭对虾的肝胰腺及前中肠的上皮细胞而引起组织坏死。 相似文献
4.
利用组织切片法研究了斑节对虾杆状病毒（MBV）在养殖早期班节对虾及养成期斑节对虾中肠腺各部位的分布特点。斑节对虾的中肠腺可分为头部、亚头部、中部、亚尾部和尾部等5个部分，其上皮细胞包括E细胞、F细胞、B细胞和R细胞等4种类型。头部和尾部均以E细胞为主、亚头部、中部和亚尾部均以R细胞为主。在感染度较轻的样本中，MBV一般只感染F细胞、R细胞和B细胞。感染度较重时，E细胞也同时受累。在养殖早期虾和养成期虾中，MBV相对感染度在中肠腺各部分的分布均以中部最高，而且以中部的中间最高，离中部越远，相对感染度越低。 相似文献
5.
对虾皮下及造血组织坏死杆状病毒单克隆抗体的研制 总被引：7，自引：1，他引：7
用经密度梯度离心纯化的对虾皮下及造血组织坏死杆状病毒（ＨＨＮＢＶ—９３７）注射Ｂａｌｂ／ｃ小鼠，取其免疫后的脾细胞与ｓｐ２／０骨髓瘤细胞进行融合。经筛选与克隆，获得７株抗ＨＨＮＢＶ的单克隆杂交瘤细胞株。其中Ｆ９株的染色体数为８７，ＥＬＩＳＡ测定培养上清中单抗滴度为１∶８，腹水为１∶１０００。患有杆状病毒的皮下及造血组织坏死症的对虾组织切片的单抗酶联免疫染色，表明Ｆ９株单抗只与ＨＨＮＢＶ病灶发生特异性结合，与对虾组织结构不发生特异性反应。将其用于现场样品的检测研究表明，该抗体适于用来进行对虾暴发性流行病的病原检测。 相似文献
6.
本文比较观察了不同规格的健康和患白便综合征凡纳滨对虾Litopenaeus vannamei肝胰腺、中肠、后肠、后盲囊等的组织结构。组织病理显示:白便综合征凡纳滨对虾肠道严重病变,肠腔空,环肌两侧增生大量成纤维细胞,这些增生细胞取代了肠上皮,而不见正常柱状上皮细胞;肝胰腺有不同程度和性质的病变,如腺管萎缩、崩解、坏死等。凡纳滨对虾患白便综合征的组织病变演变过程为:最初肠上皮基膜下出现一圈增生细胞,肠上皮柱状细胞脱离基膜;增生细胞持续增生增多,肠上皮崩解脱落至肠腔,增生细胞完全取代肠上皮;增生细胞不断增生,增生细胞层逐渐变厚,随着时间的推移,增生细胞层近腔端的一圈增生细胞坏死,出现一圈棕黄色物质,最终坏死细胞连同棕黄色物质脱落至肠腔。脱落至肠腔的腺管细胞、上皮细胞、增生细胞排出体外即为肉眼可见的白便。 相似文献
7.
中国对虾血淋巴抗凝剂的筛选 总被引：11，自引：3，他引：11
依据中国对虾血细胞电镜照片中细胞整体形态和细胞器、细胞核的变化,研究对虾抗凝剂成分配比,初步确认了一种理想的抗凝剂配方NaCl450mM,KCl100mM,EDTA,Na210mM,HEPES10mM(pH7.45)。它能够良好的保持细胞的整体形态以及细胞器、细胞核的形态。 相似文献
8.
本文报道１９９４年－１９９５年大连地区由杆状病毒引起的自然发病和人工感染实验中国对虾组织病理学研究。结果表明，自然发病虾和人工感染的虾组织病理相同，在胃、皮下、类淋巴、造血组织、鳃、中肠、后肠、肝胰腺、心脏等组织器官的上皮细胞、干细胞和结缔组织细胞中都可观察到显著胀大，呈空泡状或被苏木精浓染的细胞核。胃是病理变化出现最早且最严重的器官。在电镜下，胃、甲壳下上皮细胞核中可见大量长２７０～３５０ｎｍ，宽１００～１２０ｎｍ的杆状病毒颗粒，细胞核、线粒体出现胀大，嵴断裂、消失情况。干细胞、上皮细胞和疏松结缔组织细胞为靶细胞。 相似文献
9.
《水产科学》2019,(6)
随着对虾养殖产业集约化程度不断提高,对虾病害的发生越来越多,成为制约产业发展的主要因素。自江苏南部沿海滩涂沉积物中分离得到252株细菌,以14株凡纳滨对虾急性肝胰腺坏死综合征致病性副溶血弧菌为指示菌,经过初筛和复筛,得到1株拮抗效果较好的菌株H0011;经形态、生理生化特性和16S rDNA基因序列鉴定,确定菌株H0011为普罗威登斯菌。利用普罗威登斯菌H0011对凡纳滨对虾进行高密度胁迫试验测试其安全性,试验期间浸浴组和投喂组对虾状态良好,无发病和死亡现象。对普罗威登斯菌H0011的生长条件优化后,最优生长温度为30℃,盐度为10,pH为8。普罗威登斯菌H0011 4 h进入对数生长期,20 h达到生长高峰。本研究获得了对凡纳滨对虾急性肝胰腺坏死综合征具有防控作用的高效拮抗作用菌,将为对虾急性肝胰腺坏死综合征的防控提供新的思路。 相似文献
10.
脊尾白虾杆状病毒病研究 总被引：1，自引：0，他引：1
脊尾白虾是一种野生的经济虾类，广温，广盐，杂食性，分布面极广，繁殖季节长。我们已在其病虾的肌肉，中肠等组织中发现Ｃ亚群杆状病毒，其形态结构及宿主的细胞病理学特征均与引起中国对虾，长毛对虾，日本对虾等养殖对虾病毒性流行病的Ｃ亚群杆状病毒类同。由此认为，脊尾白虾是我国养殖对虾病毒性流行病病原的常年媒介体。 相似文献
11.
Y-S Lai S Murali H-C Chiu H-Y Ju Y-S Lin S-C Chen I-C Guo K Fang & C-Y Chang 《Journal of fish diseases》2001,24(5):299-309
A nodavirus was isolated from diseased yellow grouper, Epinephelus awoara , larvae cultured in southern Taiwan. The histopathology and RT–PCR results confirmed that it was a fish nodavirus; its coat protein gene sequence was similar to that of red spotted grouper nervous necrosis virus (RGNNV) and it is named yellow grouper nervous necrosis virus (YGNNV). A new nodavirus-susceptible cell line, grouper brain (GB) was established and characterized from the brain tissue of yellow grouper. The GB cells multiplied well in Leibovitz's L-15 medium supplemented with 10% foetal bovine serum at temperatures between 24 and 32 °C, and have been subcultured more than 80 times, becoming a continuous cell line. The GB cell line consists of fibroblast-like cells and some epithelioid cells. The cell line yielded titres of YGNNV up to 108.5 TCID50 mL–1 . The GB cells effectively replicated the virus at 28 °C, which could be purified to homogeneity by caesium chloride gradient centrifugation. Electron microscopy studies showed that purified virus particles were 25–30 nm in diameter. The cytoplasm of infected cells was filled with aggregates of virus particles. These results indicate that the GB cell line is a significant tool for the study of fish nodaviruses. 相似文献
12.
Two new cell cultures from flounder, Paralichthys olivaceus (Temminck & Schlegel), flounder fin (FFN) cells from fin tissue and flounder spleen (FSP) cells from spleen tissue, were established and characterized. The cells multiplied well in Eagle's minimum essential medium, supplemented with 10% foetal bovine serum, and have been subcultured more than 100 times, becoming continuous cell lines. Modal diploid chromosome number of FFN and FSP cells was 64 and 62, respectively. Polymerase chain reaction products were obtained from FFN and FSP cells with primer sets ofmicrosatellite markers of flounder. Optimal growth temperature was 20 degrees C and consisted of epithelioid cells. FFN and FSP cells showed cytopathic effects after inoculation of infectious pancreatic necrosis virus, marine birnavirus, chum salmon virus, infectious haematopoietic necrosis virus, spring viraemia of carp virus and hirame rhabdovirus. Thus these new cell lines may be useful for studying a wide range of fish viruses. 相似文献
13.
Yuan Zheng Na Wang Ming-Shu Xie Zhen-Xia Sha Song-Lin Chen 《Fish physiology and biochemistry》2012,38(6):1635-1643
A new cell line (TSHKC) derived from half-smooth tongue sole (Cynoglossus semilaevis) head kidney was developed. The cell line was subcultured for 40 passages over a period of 360?days. The cell line was optimally maintained in minimum essential medium supplemented with HEPES, antibiotics, fetal bovine serum, 2-Mercaptoethanol (2-Me), sodium pyruvate and basic fibroblast growth factor. The suitable growth temperature for TSHKC cells was 24?°C, and microscopically, TSHKC cells were composed of fibroblast-like cells. Chromosome analysis revealed that the TSHKC cell line had a normal diploid karyotype with 2n?=?42, contained the heterogametic W chromosome. The TSHKC cell line was found to be susceptible to lymphocystis disease virus. The fluorescent signals were observed in TSHKC when the cells were transfected with green fluorescent protein and red fluorescent protein reporter plasmids. 相似文献
14.
This study established and characterized a new cell line (MAF) from the fin of blunt snout bream (Megalobrama amblycephala), a freshwater fish cultivated in China. MAF cells proliferated well in medium 199 supplemented with 10 % fetal bovine serum at 28 °C and have been subcultured more than 95 times in almost a year. MAF cells were revived at 90–95 % viability after 3–6 months of storage in liquid nitrogen. Karyotyping indicated that the modal chromosome number of MAF cells was 48. The MAF cell line consisted predominantly of fibroblastic and epithelial-like cells from M. amblycephala, which was confirmed by immunofluorescence and mitochondrial 12s rRNA sequencing. Viral susceptibility tests showed that MAF cells were susceptible to infection by snakehead rhabdovirus, spring viremia carp virus, and channel catfish virus, which was demonstrated by the presence of cytopathic effect, high viral titers, and PCR products. Bacterial cytotoxicity studies showed that extracellular products from Aeromonas hydrophila were toxic to MAF cells. Cu2+ was also cytotoxic to MAF cells, and the 24-h IC50 value was 144.48 μmol/l. When MAF cells were transfected with pEGFP-N1 plasmid, bright fluorescent signals were observed, and the transfection efficiency reached up to 5 %. These results suggest that the MAF cell line may provide a valuable tool for studying virus pathogenesis, as well as cytotoxicity testing and genetic manipulation studies. 相似文献
15.
16.
A new continuous cell line (GF-1) was established and characterized. The GF-1 cell line, derived from the fin tissue of a grouper, Epinephelus coioides (Hamilton), was maintained in L15 medium containing 5% foetal bovine serum (FBS) at 28 °C, and has been subcultured more than 160 times since 1995. The majority of GF-1 cells are fibroblast-like, together with some epithelioid cells. Spontaneous transformation of GF-1 cells occurred during subculture 50 to subculture 80, and led to an increase of plating efficiency, less requirement of FBS and de novo susceptibility to grouper nervous necrosis virus (GNNV). Cytopathic effects (CPEs) could be observed in GF-1 cells 3–5 days post-infection with pancreatic necrosis virus (IPNV), hard clam reovirus (HCRV), eel herpes virus Formosa (EHVF) and GNNV. In addition, abundant GNNV particles were found in the cytoplasm of GNNV-infected GF-1 cells using electron microscopy and nucleic acids of GNNV virus were detected by polymerase chain reaction in the culture medium of GNNV-infected cells after CPE appeared. The experimental results indicated that GF-1 can effectively proliferate fish nodavirus and is a promising tool for studying fish nodavirus. 相似文献
17.
Y-S Lai S Murali H-Y Ju M-F Wu I-C Guo S-C Chen K Fang C-Y Chang 《Journal of fish diseases》2000,23(6):379-388
Two iridovirus-susceptible cell lines were established and characterized from grouper Epinephelus awoara kidney and liver tissues. These cell lines have been designated GK and GL, respectively. The cells multiplied well in Leibovitz's L-15 medium, supplemented with 10% foetal bovine serum, at temperatures between 20 and 32 °C, and have been subcultured more than 120 times, becoming continuous cell lines. The cell lines consist of a heterogeneous mixture of fibroblastic and epithelial cells. The viability of cells, stored frozen in liquid nitrogen (−196 °C), was 95% after 1 year. Chromosome morphologies of GK and GL cells were homogeneous. Both cell lines were susceptible to grouper iridovirus, and yielded high titres of up to 108 TCID50 mL−1 . In addition, both cell lines effectively replicated the virus, which could be purified to homogeneity by cesium chloride gradient centrifugation. Electron microscopy studies showed that purified virus particles were 170±10 nm in diameter, and were hexagonal in shape. Virus-infected cells showed an abundance of virus particles inside the cytoplasm. These results show that the GK and GL cell lines effectively replicate grouper iridovirus, and can be used as a tool for studying fish iridoviruses. 相似文献
18.
The establishment and partial characterization of a continuous cell line from the dorsal fin of red sea bream, Pagrus major, are described. The cell line, designated RSBF‐2, has been subcultured for more than 100 passages since its initiation in November 2000. It was optimally maintained at 28 °C in Leibovitz L‐15 medium with 10% foetal bovine serum. Propagation of RSBF‐2 cells was serum dependent and exhibited low plating efficiency (<1.7%). Aside from long‐term cryopreservation, the cells could also be kept at 4 °C for 72 days. The distribution of the chromosome number was 38–98 with a mode of 48. The RSBF‐2 cell line was susceptible to red sea bream iridovirus but only produced a few rounded and refractory cells. Virus‐inoculated RSBF‐2 cells were then subcultured to generate a persistently infected cell line. RSBF‐2 was also very sensitive to the extracellular products of Photobacterium damselae ssp. piscicida and produced significant fluorescent signals after transfection with pEGFP‐C3. Analysis of mitochondrial cytochrome b gene sequences revealed 99% identity between the cell line and Pagrus major. 相似文献
19.
N. Wang X. L. Wang Z. X. Sha Y. S. Tian S. L. Chen 《Fish physiology and biochemistry》2010,36(4):1227-1234
A new marine fish cell line, TK, derived from turbot (Scophthalmus maximus) kidney, was established by the method of trypsin digestion and subcultured for more than 50 passages over a period of 300 days.
The TK cells were maintained in Minimum Essential Medium Eagle (MEM) supplemented with HEPES, antibiotics, fetal bovine serum
(FBS), 2-Mercaptoethanol (2-Me), and basic fibroblast growth factor (bFGF). The suitable growth temperature for TK cells was
24°C, and microscopically, TK cells were composed of fibroblast-like cells. Chromosome analysis revealed that the TK cell
line has a normal diploid karyotype with 2n = 44. Two fish viruses LCDV-C (lymphocystis disease virus from China) and TRBIV (turbot reddish body iridovirus) were used
to determine the virus susceptibility of TK cell line. The TK cell line was found to be susceptible to TRBIV, and the infection
was confirmed by cytopathic effect (CPE) and transmission electron microscopy, which detected the viral particles in the cytoplasm
of virus-infected cells. Finally, significant green fluorescent signals were observed when the TK cells were transfected with
pEGFP-N3 vector, indicating its potential utility for fish virus study and genetic manipulation. 相似文献
20.
Y‐S Lai P P Chiou W‐J Chen Y‐C Chen C‐W Chen I‐S Chiu S‐D Chen Y‐H Cheng C‐Y Chang 《Journal of fish diseases》2008,31(11):825-834
Two new cell lines have been established from the muscle and swim bladder tissues of barramundi, Lates calcarifer, and designated as BM (barramundi muscle) and BSB (barramundi swimbladder), respectively. The cells multiplied well at 28 °C in Leibovitz’s L‐15 medium supplemented with 10% foetal bovine serum, and have been continuously subcultured more than 100 times to date. Morphologically, BM cells were mostly fibroblastic, whereas BSB were mostly epithelial. Both cell lines were susceptible to grouper iridovirus (GIV) and displayed characteristics of apoptosis after viral infection. The induction of apoptosis was further assayed in GIV‐infected BM and BSB cells by various methods. The inhibition of cell growth by GIV was demonstrated by MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide] assay. Morphological observations revealed typical apoptotic features in the infected cells, including cell shrinkage and rounding, chromosome condensation and formation of apoptotic body‐like vesicles. Chromosome fragmentation was detected by DNA laddering and TUNEL assays. Finally, the appearance of phosphotidylserine on the outer leaflet of apoptotic cell membranes was confirmed by annexin V staining. This is the first report of apoptosis induced by GIV in fish cells. 相似文献