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1.
中国对虾对白斑综合征病毒的类免疫反应与验证   总被引:3,自引:0,他引:3  
对中国对虾(Fenneropenaeus chinensis)能否产生类免疫保护作用进行了初步探讨。将健康的中国对虾接种经不同辐照强度γ射线灭活的白斑综合征病毒(white wpot wyndrome virus,WSSV),连续投喂接种20 d后,对其存活个体进行抗WSSV性能测试,检验其存活情况。结果显示,至216 h实验组(射线辐照剂量分别为5 kGy、10 kGy、15 kGy和20 kGy)存活率分别为45.5%、54.2%、68%和65.4%,存活对虾的病毒携带量分别为(14.0±11.9)、(14.1±4.3)、(24.5±15.0)和(15.1±7.8)copies/ng DNA,死亡对虾病毒携带量分别为(2.1±2.0)×105、(5.2±4.4)×105、(3.8±4.4)×105和(6.2±7.7)×105copies/ng DNA,阳性对照组存活率为20%,其存活和死亡对虾病毒携带量分别为(24.7±7.4)×105copies/ng和(5.8±6.9)×105copies/ng DNA。以上结果说明,中国对虾可能存在类免疫保护作用,而且通过接种经γ射线灭活的WSSV能够提高对虾后续抗感染能力。研究结果旨为采用免疫学手段预防对虾白斑综合征提供参考。  相似文献   

2.
本研究分别对中国对虾野生群体(Wild-Fenneropenaeus chinensis, W-Fc)、中国对虾选育群体‘黄海2号’(Selected-Fenneropenaeus chinensis, S-Fc)和凡纳滨对虾商业一代苗种(Commercial- Litopenaeus vannamei, C-Lv)采用单尾定量口饲感染白斑综合征病毒(WSSV),比较W-Fc、S-Fc及C-Lv对WSSV的敏感性差异。结果显示,感染同等含量WSSV后,W-Fc、S-Fc和C-Lv的平均存活时间分别为(124.11±39.49) h、(166.79±51.54) h和(136.90±41.99) h,3组对虾间的平均存活时间存在显著性差异(P<0.05)。3组对虾在感染期间的死亡趋势:W-Fc在96 h达到死亡高峰,并且一直持续到216 h;S-Fc和C-Lv在144 h出现死亡高峰。另外,分别在感染后的3、6、12、24、36、48、72、144 h共8个时间点对3组对虾进行活体取样,利用实时荧光定量RT-PCR技术对其进行了病毒载量检测,从对虾存活时间和体内肌肉组织病毒载量的角度比较不同对虾抗病性能的差异,结果如下:48 h时,W-Fc、S-Fc和C-Lv3对虾体内肌肉组织的病毒载量分别为(1.22×106±6.14×105)、(7.10×103±7.26×102)和(1.50×104± 4.19×103) copies/ng DNA;144 h时,3组对虾体内肌肉组织病毒载量分别为(8.44×106±1.25×106)、(3.21×106±8.21×105)和(1.49×106±6.59×105) copies/ng DNA。实验结果显示,3组对虾对WSSV敏感性从高到低依次为中国对虾野生群体、凡纳滨对虾商业苗种、中国对虾选育群体,表明中国对虾选育群体‘黄海2号’在人工感染WSSV条件下表现出了良好的抗病性能。  相似文献   

3.
斑节对虾血淋巴细胞对鳗弧菌的清除作用   总被引:2,自引:0,他引:2       下载免费PDF全文
通过检测注入斑节对虾(Penaeus monodon)血淋巴内的鳗弧菌(Vibrio anguillarumH)浓度、血淋巴细胞浓度和血淋巴细胞的组成变化,研究斑节对虾血淋巴细胞对进入体内细菌的清除作用。实验显示,斑节对虾能够迅速清除注入其体内的鳗弧菌。注射鳗弧菌悬液(107~108CFU/mL)5 min后,血淋巴中可检测到的鳗弧菌浓度是(1.4±0.6)×106CFU/mL,而2 h后所检测到的鳗弧菌的浓度仅相当于前者的3.1%。7天后,仅在部分个体的血淋巴中检测到少量鳗弧菌。伴随着血淋巴液内鳗弧菌的减少,对虾的血淋巴细胞浓度发生变化。鳗弧菌注入对虾体内5 min后,血淋巴细胞浓度为(3.51±1.69)×107/mL,注射后2 h时达到最低值,为(2.39±1.76)×107/mL,然后逐步恢复,注射后48 h达到(3.97±1.60)×107/mL。对照组血淋巴细胞浓度起初略微降低,然后逐步增加,在2 h达到最大值后又逐渐降低;除注射后48 h外,对照组血淋巴细胞浓度始终高于实验组。鳗弧菌注射入斑节对虾体内后,不同的细胞种类呈现出不同的变化规律:在注射后2 h,对照组透明细胞占总细胞数的比例显著高于实验组,对照组半颗粒细胞的含量低于实验组。注射鳗弧菌后10 min,血淋巴中颗粒细胞的相对含量显著降低,此时实验组的颗粒细胞相对含量也低于对照组。而对于降解细胞细胞核的相对含量,实验组和对照组表现出了显著不同的变化趋势。在对照组中,24 h内其含量一直在降低,而后恢复;而对照组却在注射10 min时达到最高值,而后遵循对照组类似的变化。[中国水产科学,2006,13(1):28-32]  相似文献   

4.
本研究分别对凡纳滨对虾"壬海1号"(Litopenaeus vannamei"Renhai No.1")和中国明对虾"黄海2号"(Fenneropenaeus chinensis"Huanghai No.2")在3种温度条件下(24℃、28℃和32℃),采用单尾定量口饲感染白斑综合征病毒(White spot syndrome virus,WSSV)的方法进行感染实验,比较2种对虾在不同温度情况下对WSSV的耐受性差异(L代表凡纳滨对虾,F代表中国明对虾)。结果显示,L-24℃和F-24℃组的平均存活时间分为(184.05±69.56)h和(101.68±38.45)h;L-28℃和F-28℃组的平均存活时间分别为(100.25±26.79)h和(73.38±22.22)h,相同温度组内均存在显著性差异(P0.05);截至第15天,L-32℃和F-32℃组的存活率分别为45.74%和23.47%。3个温度组对虾在50%的死亡率时的存活时间分别为178 h和98 h、98 h和74 h、292 h和78 h;死亡高峰时间分别为第5天和第4天、第5天和第4天、第10天和第4天。另外,分别在感染后的12 h、1 d、2 d、3 d、4 d、5 d、6 d、7 d、15 d共9个时间点对每组对虾进行活体取样,利用实时荧光定量RT-PCR技术对其进行病毒载量检测,从对虾体内肌肉组织病毒载量的角度探寻不同对虾抗病性能的差异。6 d时,L-24℃和F-24℃组对虾肌肉内病毒载量分别达到(2.97×10~6±7.44×10~6)和(8.08×10~6±3.22×10~6)copies/ng DNA,差异极显著(P0.01),L-28℃和F-28℃组分别达到(6.73×10~6±1.49×10~6)和(1.20×10~7±6.15×10~5)copies/ng DNA,差异极显著(P0.01);15 d,L-32℃和F-32℃组分别达到(5.18×10~4±4.32×10~4)和(3.78×10~4±8.97×10~4)copies/ng DNA,差异显著(P0.05)。研究表明,2种对虾在3种温度环境下感染WSSV后,凡纳滨对虾耐受WSSV能力要高于中国明对虾;不同温度下同种对虾肌肉体内WSSV的增殖能力从强到弱依次为28℃组、24℃组和32℃组。  相似文献   

5.
本研究分别对中国对虾野生群体(Wild-Fenneropenaeus chinensis,W-Fc)、中国对虾选育群体‘黄海2号’(Selected-Fenneropenaeus chinensis,S-Fc)和凡纳滨对虾商业一代苗种(CommercialLitopenaeus vannamei,C-Lv)采用单尾定量口饲感染白斑综合征病毒(WSSV),比较W-Fc、S-Fc及C-Lv对WSSV的敏感性差异。结果显示,感染同等含量WSSV后,W-Fc、S-Fc和C-Lv的平均存活时间分别为(124.11±39.49)h、(166.79±51.54)h和(136.90±41.99)h,3组对虾间的平均存活时间存在显著性差异(P0.05)。3组对虾在感染期间的死亡趋势:W-Fc在96 h达到死亡高峰,并且一直持续到216 h;S-Fc和C-Lv在144 h出现死亡高峰。另外,分别在感染后的3、6、12、24、36、48、72、144 h共8个时间点对3组对虾进行活体取样,利用实时荧光定量RT-PCR技术对其进行了病毒载量检测,从对虾存活时间和体内肌肉组织病毒载量的角度比较不同对虾抗病性能的差异,结果如下:48 h时,W-Fc、S-Fc和C-Lv3对虾体内肌肉组织的病毒载量分别为(1.22×10~6±6.14×10~5)、(7.10×10~3±7.26×10~2)和(1.50×10~4±4.19×10~3)copies/ng DNA;144 h时,3组对虾体内肌肉组织病毒载量分别为(8.44×10~6±1.25×10~6)、(3.21×10~6±8.21×10~5)和(1.49×10~6±6.59×10~5)copies/ng DNA。实验结果显示,3组对虾对WSSV敏感性从高到低依次为中国对虾野生群体、凡纳滨对虾商业苗种、中国对虾选育群体,表明中国对虾选育群体‘黄海2号’在人工感染WSSV条件下表现出了良好的抗病性能。  相似文献   

6.
氨氮胁迫下白斑综合征病毒对凡纳滨对虾的致病性   总被引:1,自引:0,他引:1  
为了评价养殖水环境中氨氮(NH_4-N)对凡纳滨对虾(Litopenaeus vannamei)的危害性,开展了NH_4-N胁迫对凡纳滨对虾感染白斑综合征病毒(WSSV)后的死亡率、WSSV增殖速率和对虾主要免疫相关酶活性影响的实验。在NH_4-N胁迫质量浓度为15.6 mg·L-1,分别注射2×105和2×106个WSSV粒子,结果显示,NH_4-N胁迫下注射2×105个WSSV粒子的凡纳滨对虾第144小时死亡率达到53.3%,显著高于无胁迫组(40.0%)。对虾鳃组织WSSV荧光定量PCR检测结果显示,NH_4-N胁迫下凡纳滨对虾鳃组织内WSSV的增殖加快。此外,免疫相关酶活性结果显示,NH_4-N浓度突变会促使对虾血清中酚氧化酶(PO)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性短暂升高后持续降低。由此可见,NH_4-N胁迫会加快WSSV在患病凡纳滨对虾体内的增殖,导致更高死亡率,这可能是因为胁迫造成了对虾免疫相关酶活性降低和抗病原感染能力下降。  相似文献   

7.
白斑综合征病毒感染与对虾的免疫防御反应   总被引:1,自引:1,他引:1  
对虾白斑综合征病毒(WSSV)感染对虾后,最典型的免疫反应是对虾开放循环系统的血淋巴细胞数量急剧下降,血淋巴凝结功能下降,感染部位聚集了大量的血淋巴细胞,且以颗粒细胞为主。WSSV可感染对虾颗粒细胞和小颗粒细胞,其中小颗粒细胞感染率高、感染速率快,感染后大颗粒细胞占血细胞总数的比例可增加到50%;血淋巴中的总糖、总碳水化合物、总蛋白和游离氨基酸显著提高,超氧化物歧化酶、过氧化氢酶和诱导性一氧化氮合成酶的活性显著降低。自然状态下广泛存在WSSV潜伏感染,潜伏感染的存在会导致存在免疫反应情况下的感染复发,并且有助于病毒的传播;不同WSSV感染状态下过氧化物(POD)差异显著,其平均值由大到小依次为:潜伏感染虾样、中度感染虾样、严重感染虾样。而其抗菌活性(UA)、溶菌活性(UL)、酶氧化酶活性(PO)、碱性磷酸酶(ALP)和凝集效价(HAT)差异不显著;潜伏感染个体对再次接种WSSV有“类免疫应答”的抗性,这种抗性不是来源于发病期对虾的天然抗性,而是WSSV感染后的一种免疫系统增强。宿主细胞凋亡可能是感染对虾在高温时反而维持较高成活率的主要机制。免疫增强剂可对对虾防御WSSV感染产生影响,脂多糖、葡聚糖、肽聚糖、岩藻依聚糖和双链核糖核酸都已被证实可提高对虾抗病毒感染的免疫保护。WSSV主要囊膜蛋白VP28可诱导对虾对WSSV感染产生抗性降低累积死亡率,高效价的病毒抗血清具有良好的保护作用;对虾抗菌肽也可通过抑制病毒的复制而起保护作用。  相似文献   

8.
以超低温保存的感染了白斑综合征病毒(WSSV)的中国对虾制备的病毒粗提液为毒种,注射感染凡纳对虾并收集濒死虾,DNA斑点杂交检测每尾凡纳对虾WSSV感染状况。取DNA斑点杂交呈强阳性的30尾对虾,平均分为3组,病毒粗提液也平均分成3组,3组材料分别通过^60Co辐照,辐照时间分别为12、24和36h,辐照剂量为0.8KGy/h。辐照后的材料经PCR检测证实^60Co辐照不能完全破坏WSSV的DNA组成。以辐照后的感染白斑综合征病毒的对虾个体和白斑综合征病毒粗提液为感染毒种,人工感染健康凡纳对虾,验证^60Co辐照对病毒感染力的破坏作用,证实^60Co辐照可显著降低WSSV病毒粗提液的感染力,^60Co辐照可适当降低WSSV感染对虾的感染力。  相似文献   

9.
用含3×103拷贝·g^-1、6×1062拷贝·g^-1和2×10^2拷贝·g^-1的对虾白斑综合征病毒(white spot syndrome virus,WSSV)粗提液和PBS液(对照)注射感染病毒携带量约1×10^5拷贝·g^-1的斑节对虾(Penaeusmonodon),分别于第15分钟、第30分钟、第1小时、第3小时、第6小时、第12小时、第24小时、第48小时、第72小时取样,研究了WSSV感染对斑节对虾血清内酸性磷酸酶(acidphosphatase,ACP)、碱性磷酸酶(alkalinephospha—tase,AKP)、酚氧化酶(phenoloxidase,PO)、过氧化物酶(peroxidase,POD)和超氧化物歧化酶(superoxide dismutase,SOD)活性的影响。结果表明,在3种感染浓度下ACP、AKP、SOD活性总体呈现先上升后下降随后上升的趋势,其中SOD活性后期水平显著高于初期;PO、POD活性总体呈现先下降后上升随后下降最后上升的趋势,但PO后期活性水平与初期相当,而POD后期活性水平显著高于初期。各免疫相关酶的反应强度与WSSV的感染浓度存在一定关系,除ACP外其余4种酶的活性变化均以6×10^2拷贝·mL^-1浓度组最为敏感。PBS组5种免疫酶活性变化幅度均显著小于3种WSSV浓度感染组。  相似文献   

10.
刘强  周俊芳  房文红  王元  李新苍  周帅 《水产学报》2013,37(7):1106-1112
为了评价养殖水环境中毒死蜱对凡纳滨对虾生存的危害性,开展了毒死蜱胁迫下白斑综合征病毒(WSSV)对凡纳滨对虾致死实验,分析了毒死蜱胁迫下凡纳滨对虾鳃组织WSSV含量和肌肉组织乙酰胆碱酯酶活性变化。通过急性毒性实验测定了毒死蜱对凡纳滨对虾的半致死浓度(LC50),随着暴露时间的延长,LC50值显著下降,存在着浓度-反应的正向关系,96 h LC50为0.758 μg/L(0.521~0.987 μg/L)。在此基础上,确定了毒死蜱胁迫实验浓度为0.2 μg/L,此浓度下药浴4 d后对凡纳滨对虾注射WSSV,结果显示:毒死蜱胁迫下注射WSSV组的对虾死亡率(83.33?Ee4.7%)极显著高于乙醇-WSSV组(40.00?Ee0.9%);对虾鳃组织WSSV荧光定量PCR检测结果显示:感染72 h后,毒死蜱-WSSV组WSSV含量约是乙醇-WSSV组的4倍;感染96 h后,毒死蜱-WSSV组WSSV含量显著增加,约是72 h毒死蜱-WSSV组的4.9倍,是96 h乙醇-WSSV组的5.9倍。毒死蜱胁迫下,对虾肌肉组织乙酰胆碱酯酶(AchE)活性低于对照组20%左右。由此可见,毒死蜱胁迫下,WSSV增殖速率加快,导致对虾死亡率升高。  相似文献   

11.
Mud crab, Scylla serrata (Forskal), is the most commercially important marine crab species in China. In recent years, serious diseases have occurred in major mud crab culture regions in SE China. PCR detection of white spot syndrome virus (WSSV) in diseased mud crabs collected from Zhejiang Province during 2006–2008 showed a prevalence of 34.82%. To study the pathogenicity of WSSV to mud crab, healthy mud crabs were injected intramuscularly with serial 10‐fold dilutions of a WSSV inoculum. The cumulative mortalities in groups challenged with 10?1, 10?2, 10?3 and 10?4 dilutions were 100%, 100%, 66.7% and 38.9% at 10 days post‐injection, respectively. All moribund and dead mud crabs except the control group were positive for WSSV by PCR. Based on the viral load of the WSSV inoculum by quantitative real‐time PCR, the median lethal dose (LD50) of WSSV in S. serrata was calculated as 1.10 × 106 virus copies/crab, or 7.34 × 103 virus copies g?1 crab weight. The phenoloxidase, peroxidase and superoxide dismutase activities in haemolymph of WSSV‐infected moribund crabs, were significantly lower than the control group, whereas alkaline phosphatase, glutamate‐pyruvate transaminase and glutamic‐oxaloacetic transaminase were higher than in the control group. WSSV was mainly distributed in gills, subcuticular epithelia, heart, intestine and stomach as shown by immunohistochemical analysis with Mabs against WSSV. The epithelial cells of infected gill showed hypertrophied nuclei with basophilic inclusions. Numerous bacilliform virus particles were observed in nuclei of infected gill cells by transmission electron microscopy. It is concluded that WSSV is a major pathogen of mud crab with high pathogenicity.  相似文献   

12.
White spot disease caused by white spot syndrome virus (WSSV) is the major issue of huge economic destruction globally in the shrimp aquaculture industry. In the present investigation, WSSV prevalence associated with disease resistance was studied among wild black tiger shrimp, Penaeus monodon (Fabricius) from four distant geographic locations along the East coast of India during 2009–2010. Results suggested that the WSSV prevalence in wild P. monodon was the highest (56.2%) in Chennai, Tamil Nadu followed by Digha, West Bengal (10.9%), Visakhapatnam, Andhra Pradesh (0.6%) and Chilika, Orissa (0%). Quantitative data suggested that the mean copy number of WSSV among these four places was 1.4 × 106, 4.6 × 104, 1.6 × 102 and 2.3 × 102 copies μg?1 shrimp genomic DNA respectively. The disease resistant prevalence using the 71 bp microsatellite DNA marker was the highest among Chilika, Orissa (63.6%) and Visakhapatnam, Andhra Pradesh (63.5%). Higher WSSV prevalence in Chennai, Tamil Nadu and Digha, West Bengal corresponded to lower disease resistant prevalence (24% and 40.2%). Conclusively, probably collection of broodstock of P. monodon from places like Chilika and Visakhapatnam would be a much safer approach for the development of specific pathogen‐resistant shrimp aquaculture.  相似文献   

13.
The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 103 to 3.76 × 105 cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 105 cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

14.
白斑综合征病毒(WSSV)3种PCR检测方法的灵敏度比较   总被引:1,自引:0,他引:1  
为了探讨不同PCR检测方法的灵敏度,分别利用TaqMan实时定量PCR、世界动物卫生组织(OIE)公布的巢式PCR引物(简称OIE)、黄海水产研究所种质资源与工程育种研究室(GB)设计的引物(简称GB)及2种巢式PCR对应的一步法PCR,对具有不同白斑综合征病毒(White Spot Syndrome Virus,WSSV)含量的中国明对虾(Fenneropenaeus chinensis)样品进行检测.结果显示,当使用已知病毒含量的标准品进行检测时,TaqMan实时定量PCR方法可以检测到l0个WSSV拷贝;OIE巢式PCR与GB巢式PCR方法分别可检测到104和103个WSSV拷贝;单独使用OIE巢式PCR的外引物和内引物扩增时,分别可检测到5×104和2.5×104个WSSV拷贝;单独使用GB巢式PCR的外引物和内引物进行一步法PCR扩增时,分别可检测到104和5×103个WSSV拷贝.使用上述PCR方法分别对44份未知WSSV含量的样品进行验证,定量PCR方法检测阳性率为84.09%,OIE巢式PCR与GB巢式PCR方法检测的阳性率分别为18.18%和27.27%;单独使用OIE巢式PCR的外引物和内引物扩增检测的阳性率均为15.91%;单独使用GB巢式PCR的外引物和内引物扩增检测的阳性率分别为18.18%和20.45%.根据以上结果,PCR方法检测WSSV的灵敏度由高到低依次为:定量PCR、巢式PCR、一步法PCR.  相似文献   

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In the present study, we used TaqMan real‐time polymerase chain reaction to quantify and compare infection of white spot syndrome virus (WSSV) with shrimp production of Fenneropenaeus chinensis cultured in outdoor ponds along the west coast of the South Korea. In 2007, a total of 60 specimens in summer and 116 specimens in autumn were collected from 12 growing‐out ponds and 12 harvest ponds respectively. Pond harvest data were obtained from farmers. Of the summer samples, all specimens were WSSV positive, with a wide range of 12.4–7.0 × 107 (mean 7.5 × 106) copies ng?1 DNA; shrimp production was 1.7 metric tonnes per hectare (mt ha?1). Of the 116 autumn‐sample specimens, 81 (69.8%) were WSSV positive; WSSV infection had been decreased dramatically, to 0–7.2 (mean 3.5) copies ng?1 DNA. Shrimp production of autumn ponds was 2.1 mt ha?1. Statistical analysis indicated that the difference in WSSV infections detected in summer and autumn was highly significant (P<0.01). In summer, seven ponds (58.3%) with low‐WSSV infection loads (0–1000 WSSV copies ng?1 DNA) had shrimp production of 2.7 mt ha?1; the others had shrimp production of only 0.2 mt ha?1. The mean shrimp production between the two infection levels showed a highly statistically significant difference (P<0.01).  相似文献   

17.
Two kinds of specific chicken egg yolk immunoglobulins (IgYs), IgY‐WSSV and IgY‐VP28, were, respectively, raised against the 2 mM binary ethylenimine (BEI)‐inactivated white spot syndrome virus (WSSV) and a principal envelope protein VP28. The activity of purified specific IgYs was stable under the conditions of 20–70 °C, pH 3.0–10.0 and 0–700 g L?1 sucrose solution. In the neutralization assay, these high‐affinity IgY antibodies can specifically bind with the virus particles to protect shrimp (Fenneropenaeus chinensis) against WSSV infection. After oral delivery for 20 days, the IgY‐WSSV exerted a higher protection effect (RPS: 71.5%) than IgY‐VP28 (RPS: 63.7%). Moreover, an increase in RPS (79.2%) was found on addition of IgY‐WSSV:VP28 (0.1% IgY‐VP28 plus 0.2% IgY‐WSSV). This may indicate that neutralization of WSSV refers to the multiple‐hit model. By time‐course study of the levels of the specific IgYs in vivo, the data showed that the titre was enhanced to a relatively high level (P/N=8.35±0.45) at 3 days post administration, declined slightly (P/N=7.13±1.01) at 7 days post administration and then remained stable for further investigation. The stable antibody level potentially contributes towards blocking a large number of WSSV particles from entering and infecting on the major tissues at the early and late stages after challenge in shrimp.  相似文献   

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Determination of differentially expressed protein profile is necessary to understand the host response to viral infection. Proteomics can be applied as a tool to examine white shrimp Litopenaeus vannamei molecular responses against white spot syndrome virus (WSSV) infection, thus enabling development of effective strategies to reduce their impact on farms. In the present study, specific pathogen-free shrimp was tested against WSSV infection under several time intervals. Shrimps were submitted to a viral load of with 5.5 × 106 viral copies in 100 μL/shrimp. The monitoring of infection was performed in intervals of 6, 12, 24, 48 and 72 h after infection. The analysis was realized using 2-DE, and differentially expressed proteins were identified by MALDI-TOF mass spectrometry (MS) peptide mass fingerprint (PMF). Between the differentially expressed proteins found in the infected animals, the most important were identified as caspase-2, ubiquitin and F1-ATP synthase. They are interesting candidates for biomarkers because could be related to the beginning of apoptosis process. The differentially expressed protein profile creates a new paradigm in the analysis of L. vannamei shrimp molecular response to WSSV infection and in virus–host relationship. Furthermore, it proposes potential biomarkers that allow strategies both selecting less susceptible individuals and reducing the impact of viruses on farms.  相似文献   

20.
Quantitative real time PCR, recently developed in molecular biology, is applied in this paper to quantify the white spot syndrome virus (WSSV) in infected shrimp tissue. The WSSV content in moribund shrimp of all species tested ( Penaeus stylirostris, P. monodon, P. vannamei ) ranged from 2.0 × 104 to 9.0 × 1010 WSSV copies μg–1 of total DNA ( n =26). In whole moribund post-larvae, 4.3 × 109 WSSV copies μg–1 of DNA were detected which is equivalent to 5.7 × 1010 WSSV copies g–1 of post-larvae. The comparison of WSSV content between different tissues showed that muscle and hepatopancreas tissues contained 10 times less virus than gills, pleopods and haemolymph. With inocula of known virus content, bioassays by immersion challenge showed that a minimum of five logs of WSSV copies was necessary to establish disease in the challenged shrimp. In contrast, five logs of WSSV copies injected into shrimp muscle produced a LT-50 of 52 h. This real time polymerase chain reaction (PCR) technique is sensitive (four copies), specific (negative with DNA from shrimp baculoviruses and parvoviruses), dynamic (seven logs) and easy to perform (96 tests in <4 h).  相似文献   

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