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In this study, the possible influence of temperature on infectious pancreatic necrosis virus (IPNV)-induced apoptosis in a zebrafish liver epithelium (ZLE) cell line was investigated. At a lower temperature (18 degrees C), there was expression of viral proteins VP2 and VP3 at 4 h post-infection (p.i.). At this time no expression was found in the high temperature group at 28 degrees C. The cell survival ratio was 52 and 18% at 24 and 48 h p.i., respectively, during IPNV infection at 18 degrees C. In addition, we assayed for apoptosis in IPNV-infected cells with terminal deoxynucleotidyl transferase (TdT)-mediated end labelling (TUNEL) of DNA at different dosages of virus. We found a ratio of apoptotic cells of 8 and 25% at 12 and 18 h p.i., respectively, in the multiplicity of infection (MOI) 1 group. The MOI 10 group had 20 and 45% apoptotic cells at 12 and 18 h, respectively. Furthermore, at 18 degrees C IPNV activated the caspase-8 and 3 from 1.5 to 2 times at 12 and 18 h p.i., respectively. Taken together, these findings suggest that successful virus replication occurs at the low temperature (18 degrees C) compared with the non-permissive temperature of 28 degrees C. Thus, IPNV replication is capable of activating caspase-8 and -3 and inducing host apoptosis. 相似文献
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Qian D Shi Z Zhang S Cao Z Liu W Li L Xie Y Cambournac I Bonami JR 《Journal of fish diseases》2003,26(9):521-527
A disease of Macrobrachium rosenbergii, the giant freshwater prawn, farmed in China was recently recorded in Zhejiang, Jiangsu, Shanghai, Guangxi and Guangdong provinces. The clinical sign of the disease, which develops in post-larvae (PL), is a whitish appearance of the muscles, particularly noticeable in the abdomen. Mortalities may reach 100% in some hatcheries. Investigations by transmission electron microscopy after negative staining of diseased PL homogenates showed the presence of two types of viral particles: one, unenveloped, icosahedral in shape, 26-27 nm in diameter, the second, much smaller, about 14-16 nm in diameter, designated extra small virus particle (XSV). The large virus has a genome with two pieces of ssRNA (RNA-1 and RNA-2), of 3 and 1.2 kb, respectively. Hybridization tests confirmed that this large virus is closely related to M. rosenbergii nodavirus (MrNV) which was isolated from diseased prawns in a hatchery in the French West Indies. Its very small size and hypothesized biochemical and biological characteristics suggest XSV is a new type of crustacean virus. As XSV has always been found associated with the larger virus (nodavirus) and is located in muscle and connective cells of diseased animals, it could be an autonomous virus, a helper-type virus or a satellite-like virus. 相似文献
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White-clawed crayfish, Austropotamobius pallipes, were endemic to the Nant watershed, Ardéche, France, until they were extirpated by epizootic mortality at the beginning of the twentieth century. A. pallipes were successfully reintroduced to the Nant watershed in the middle of the twentieth century. However, epizootic mortality was observed in the Nant watershed in the summer of 2000 during which time A. pallipes was extirpated from downstream regions. Dead and moribund crayfish were again detected in several episodes in summer 2001 and by October the range of A. pallipes was reduced to the headwaters of just one of the three streams in the watershed. Water quality for the watershed in summer 2001 was appropriate for crayfish habitation. Bacteriology and mycology on A. pallipes collected during several of the mortality episodes in 2001 failed to reveal a cause. However, histopathology revealed a high occurrence of intranuclear eosinophilic inclusions in hepatopancreatocytes of A. pallipes . The nuclei were hypertrophic and contained bacilliform virions consisting of a cylindrical nucleocapsid surrounded by a trilaminar envelope. Virions in section were approximately 63 × 258 nm and nucleocapsids were approximately 52 × 225 nm. It is unclear whether the intranuclear bacilliform virus was the cause of the mortality episodes or was a contributor to a disease complex involving one or several other undetected pathogens. 相似文献
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Melena J Bayot B Betancourt I Amano Y Panchana F Alday V Calderón J Stern S Roch P Bonami JR 《Journal of fish diseases》2006,29(10):589-600
Larvae and post-larvae of Penaeus vannamei (Boone) were submitted to primary challenge with infectious hypodermal and haematopoietic necrosis virus (IHHNV) or formalin-inactivated white spot syndrome virus (WSSV). Survival rate and viral load were evaluated after secondary per os challenge with WSSV at post-larval stage 45 (PL45). Only shrimp treated with inactivated WSSV at PL35 or with IHHNV infection at nauplius 5, zoea 1 and PL22 were alive (4.7% and 4%, respectively) at 10 days post-infection (p.i.). Moreover, at 9 days p.i. there was 100% mortality in all remaining treatments, while there was 94% mortality in shrimp treated with inactivated WSSV at PL35 and 95% mortality in shrimp previously treated with IHHNV at N5, Z1 and PL22. Based on viral genome copy quantification by real-time PCR, surviving shrimp previously challenged with IHHNV at PL22 contained the lowest load of WSSV (0-1x10(3) copies microg-1 of DNA). In addition, surviving shrimp previously exposed to inactivated WSSV at PL35 also contained few WSSV (0-2x10(3) copies microg-1 of DNA). Consequently, pre-exposure to either IHHNV or inactivated WSSV resulted in slower WSSV replication and delayed mortality. This evidence suggests a protective role of IHHNV as an interfering virus, while protection obtained by inactivated WSSV might result from non-specific antiviral immune response. 相似文献
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A second type of freshwater crab reovirus has been isolated from Chinese mitten crab, Eriocheir sinensis H. Milne Edwards, in China; we named it E. sinensis reovirus (EsRV816). The negatively stained virion is a non‐enveloped icosahedral particle, 60 ± 5 nm in diameter. Its genome is composed of 10 dsRNA linear pieces exhibiting an electrophoretic pattern of 5/3/2. The largest segment (RNA‐1) was cloned and sequenced. The deduced amino acid sequence, corresponding to the RdRp of the virus, showed 26% identity with the RdRp of Operophtera brumata (L.) cypovirus 19 in the genus Cypovirus and 24% identity with RdRp of Nilaparvata lugens (Stal) reovirus in the genus Fijivirus. On the basis of its ultrastructure and physicochemical properties, this virus is quite different from other crab reoviruses, and particularly with another freshwater crab reovirus EsRV905, recently classified in a new genus Cardoreovirus. This virus (EsRV816) possesses all the characters of the members of the reoviridae family and could represent a new genus. 相似文献
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Experimental infection of European crustaceans with white spot syndrome virus (WSSV) 总被引：3，自引：1，他引：3
V Corbel Z Zuprizal C Shi Huang Sumartono J-M Arcier & J-R Bonami 《Journal of fish diseases》2001,24(7):377-382
Eight European marine and freshwater crustaceans were experimentally infected with diluted shrimp haemolymph infected with white spot syndrome virus (WSSV). Clinical signs of infection and mortalities of the animals were routinely recorded. Diagnosis was by direct transmission electron microscopy (TEM), DNA hybridization (dot-blot and in situ hybridization) using WSSV probes and by PCR using WSSV specific primers. High mortality rates were noted between 7 to 21 days post-infection for Liocarcinus depurator , Liocarcinus puber , Cancer pagurus , Astacus leptodactylus , Orconectes limosus , Palaemon adspersus and Scyllarus arctus . Mortality reached 100%, 1 week post-infection in P. adspersus . When infection was successful, direct TEM observation of haemolymph revealed characteristic viral particles of WSSV, some observed as complete virions (enveloped), others as nucleocapsids associated with envelope debris. WSSV probes showed strong positive reactions in dot-blots and by in situ hybridization in sections and specific virus DNA fragments were amplified successfully with WSSV primers. White spot syndrome virus was pathogenic for the majority of the crustaceans tested. This underlines the epizootic potential of this virus in European crustaceans. 相似文献
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Nervous necrosis virus (NNV) infection induces host cell apoptosis by an ill-understood process. We utilized a fusion between enhanced green fluorescent protein (EGFP) and the zfBcl-x(L) gene in GL-av cells to select for zfBcl-x(L) stable cell lines and to assess the effectiveness of the anti-apoptotic protein Bcl-x(L) in circumventing NNV-induced cell death. Stable EGFP and EGFP-Bcl-x(L)-expressing clones were obtained at high purity within 2.5-3 months. In the latter, the EGFP-Bcl-x(L) fusion protein (approximately 58.2 kDa, as ascertained by Western blot) was predominantly targeted to mitochondria. We assayed for apoptosis in red-spotted grouper NNV Tainan no. 1 (RGNNV TN1)-infected cells with terminal deoxynucleotidyl transferase (TdT)-mediated end labelling (TUNEL) of DNA at different virus doses. NNV infection of NNV Bcl-x(L) GL-av cell line revealed a protective effect, with a decrease in TUNEL-positive cells of 7%, 8% and 31.8% at 24, 48 and 72 h, respectively. In addition, RGNNV infection of the Bcl-x(L) GL-av cell line revealed a protective effect, with an enhanced viability of 3%, 40% and 73% at 24, 48, and 72 h, respectively. We conclude that NNV-induced apoptotic cell death can be lessened in transgenic grouper fish cells. 相似文献