共查询到19条相似文献，搜索用时 671 毫秒
1.
在建立斑节对虾实验室养殖模式的基础上．对野外采集的有白斑综合症病毒(WSSV)病典型症状的斑节对虾初步纯化其病毒．并进行WSSV的PCR检测，得到WSSV的感染用样品。对健康斑节对虾分别进行浸浴感染，投喂感染和注射感染。对感染死亡个体进行WSSA的PCR检测和细菌检测，证实WSSV感染性和致死性。浸浴感染、投喂感染和注射感染的感染量分别为4mL／L、0．2g／10g虾体、1／2稀释液0．05mL／10g虾体，死亡开始时间分别为16d、42h、28h，三种感染模式最终死亡率100％．从开始死亡到全部死亡延续时间分别为15d、82h、44h。 相似文献
2.
用6.0×104拷贝、1.2×104拷贝和6.0×103拷贝3种剂量白斑综合症病毒(WSSV)对凡纳滨对虾和斑节对虾进行人工注射感染,比较了两种对虾对WSSV敏感性的差异。结果表明,凡纳滨对虾死亡时间随病毒剂量降低而延长,斑节对虾死亡时间没有明显差异;随病毒剂量的降低,凡纳滨对虾人工注射感染后病毒复制高峰时间显著延长,斑节对虾感染后病毒复制高峰时间相同,WSSV在凡纳滨对虾体内比在斑节对虾体内复制慢。对虾携带WSSV数量最低为3.3×107拷贝.g-1,最高为4.3×108拷贝.g-1。凡纳滨对虾比斑节对虾对WSSV的抵抗性更强。 相似文献
3.
对克隆的 WSSV基因组 Xba 1.8kb片段进行序列测定 ,根据所测定的核酸序列 ,通过计算机软件分析 ,设计出一对 PCR引物。所设计的 PCR引物能从纯化 WSSV及患白斑症的对虾组织中扩增出长为 92 1bp的目的 DNA片段 ,并且能检测出 0 .2～ 0 .4μg病虾肌肉组织中的 WSSV。健康对虾、感染MBV的斑节对虾仔虾及 SINPV的扩增结果均为阴性。表明所建立的 WSSV PCR检测法灵敏而且特异。 相似文献
4.
《广东海洋大学学报》2016,(1)
对感染白斑综合征病毒(WSSV)的凡纳滨对虾(Litopenaeus vannamei)分别注射0.05和0.10μg/μL的vp28-si RNA,干扰效果实验表明,si RNA最佳浓度为0.10μg/μL。凡纳滨对虾感染WSSV 24 h时注射0.10μg/μL的vp28-si RNA,检测免疫后不同时间点的干扰效果。结果表明:在注射vp28-si RNA 6 h后,实验组与对照组WSSV病毒拷贝数差异有统计学意义(P0.05);在48 h时实验组病毒拷贝数急剧下降,对照组则保持较高水平,可较好干扰WSSV病毒复制,干扰效果持续120 h,对凡纳滨对虾的保护率为40%。凡纳滨对虾用0.10 mg/m L肽聚糖(Peptidoglycan)免疫24 h后感染WSSV,检测6、12、24 h注射vp28-si RNA的保护率。结果显示:6 h注射组的保护率为80%,12 h注射组为63.33%,24 h注射组为56.67%。凡纳滨对虾在肽聚糖和vp28-si RNA作用后,可增加其对WSSV抗感染作用,降低死亡率,干扰时间越早,对对虾的保护率越高。 相似文献
5.
《广东海洋大学学报》2015,(6)
从患病凡纳滨对虾(Litopenaeus vannamei)体内提取白斑综合征病毒(WSSV),PCR证实WSSV毒性,将病毒粗提液稀释10-2、10-3、10-4和10-5倍,攻毒实验表明,最佳攻毒剂量为10-4倍稀释液。对凡纳滨对虾(Litopenaeus vannamei)分别注射0.05、0.10、0.15 mg/m L的溶藻弧菌(Vibrio alginolyticus)肽聚糖,免疫24 h后感染WSSV,测定其免疫保护率和肝胰腺的免疫学指标,探讨溶藻弧菌肽聚糖对凡纳滨对虾抗WSSV的影响。结果表明:对虾的保护率随着肽聚糖的浓度增大而增大,与对照组相比,0.05、0.10、0.15 mg/m L组的保护率分别为36.67%、46.67%、56.67%;实验组凡纳滨对虾肝胰腺中的酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)和一氧化氮合成酶(NOS)活力与对照组相比的差异有统计学意义(P0.01)。溶藻弧菌肽聚糖可有效增强凡纳滨对虾自身的非特殊性免疫能力,提高抗WSSV的能力。 相似文献
6.
【目的】建立快速检测凡纳滨对虾野田村病毒的RT-PCR方法。【方法】根据GenBank中获得的凡纳滨对虾野田村病毒的基因序列,应用Oligo6.0软件设计合成1对特异性扩增引物,对反应条件和反应体系进行优化,建立快速检测凡纳滨对虾野田村病毒的RT-PCR方法。用该方法对感染野田村病毒的凡纳滨对虾进行RT-PCR检测。【结果】RT-PCR可扩增出与设计相符的413 bp的特异性目的条带,而对白斑症病毒(WSSV)阳性虾、对虾杆状病毒(BP)阳性虾、传染性皮下及造血器官坏死病毒(IHHNV)阳性虾、肝胰腺细小病毒(HPV)阳性虾、黄头病毒(YHV)阳性虾、传染性肌肉坏死病毒(IMNV)阳性虾、桃拉病毒(TSV)阳性虾和健康虾的扩增结果均为阴性。测序比对结果表明,该方法检测结果准确,最低可检测出约100 fg/mL的野田村病毒重组质粒DNA;用该方法对从福建、广东、海南、广西、江苏、浙江、山东等地的386份临床样品进行RT-PCR检测,共检出野田村病毒阳性样品64份。【结论】该方法可用于凡纳滨对虾野田村病毒的快速检测。 相似文献
7.
《广东海洋大学学报》2015,(4)
根据Gen Bank中公布的对虾杆状病毒(Baculovirus penaei,BP)基因片段序列,设计1对特异性检测引物,建立快速检测凡纳滨对虾(Litopenaeus vannamei)对虾杆状病毒的PCR方法。用该方法对BP阳性虾进行PCR扩增,结果得到380 bp的特异性扩增条带,与实验设计相符,而对白斑综合征病毒(WSSV)、桃拉病毒(TSV)、传染性皮下及造血器官坏死病毒(IHHNV)阳性虾和健康虾的扩增结果为阴性。测序比对结果证实,该PCR方法检测结果准确,最低可检测出约1 pg的病毒DNA。利用建立的PCR方法对来自广东、广西、福建、海南和浙江的2 722份临床病料进行检测,共检出阳性病料133份,表明该PCR方法可用于对虾杆状病毒的临床快速检测。 相似文献
8.
利用 2步 PCR检测技术对湛江和阳西地区的虾苗进行白斑综合病杆状病毒 ( WSSV)检测。在仔虾 2日龄最早检测到病毒 ,有 2 5%的虾苗带有 WSSV病毒。带病毒虾苗和不带病毒虾苗分别在不同养殖模式的养殖过程中跟踪。前者在湛江湖光镇普通虾塘跟踪养殖 ,它们在变化的环境中容易发病 ,p H、盐度和温度是重要的诱发因子 ,在养殖 50～ 60 d时发病死亡 ;后者在高位池和普通池养殖跟踪 ,它们对变化的环境有较大的适应性 ,养殖时间为 80～ 1 1 0 d,在相对优良的养殖技术条件下大部分可望养殖成功。环境中有 WSSV病原传入 ,不带病毒虾苗在养殖后期可以带有 WSSV病毒 ,出现白斑虾。跟踪的普通池有爆发病害 ,但是时间延后 ,跟踪的高位池没有爆发病害。 相似文献
9.
利用雷州半岛广泛养殖的马氏珠母贝及珍珠生产中大量废弃的珍珠贝体成熟性腺，获取贝类幼体作为饵料来培育斑节对虾苗。通过比较不同投喂方式的对虾幼体的变态率，各时期的成活率和出苗率来探讨贝类幼体替代事部分替代卤虫卵的可行性。研究结果表明，投喂珠母贝幼体的试验池比投喂卤虫幼体、骨条藻的对虾幼体早期变态率要高。而平均出苗率以投喂卤虫幼体的池最高，为４９．８％；投喂珠母贝幼体的池略低，为４０．４％；而前期仅投喂 相似文献
10.
《广东海洋大学学报》2016,(4)
根据Gen Bank中对虾肠道上皮细胞微胞子虫(Enterocytozoon hepatopenaei,EHP)的基因保守序列,设计一对特异性引物,通过优化PCR扩增条件,建立快速检测凡纳滨对虾(Litopenaeus vannamei)EHP的PCR方法。用该方法对EHP阳性虾进行PCR扩增,得到与实验设计相符的330 bp特异性扩增条带,而对白斑综合症病毒(WSSV)、桃拉病毒(TSV)、传染性皮下及造血器官坏死病毒(IHHNV)、对虾杆状病毒(BP)、"棉花虾"微孢子虫、黏孢子虫的阳性虾,以及健康虾的扩增结果为阴性。测序比对发现,PCR产物序列与Gen Bank EHP基因序列的同源性为99.8%,表明该PCR方法检测结果准确。敏感性试验表明,该方法最低可检测出约100 fg的EHP质粒DNA。用该PCR方法检测广东、广西、江苏、山东、海南等地的755份临床样品,共检出阳性样品21份。该PCR方法可用于凡纳滨对虾EHP的快速检测。 相似文献
11.
对自然发生的斑节对虾烂鳃病进行了病理组织切片观察,发现病虾由杆状细菌感染造成鳃丝萎缩、坏死,鳃小片呼吸上皮脱落,鳃血中有菌团存在,而且病虾的肝胰脏、胃、肠、心脏等器官组织受毒素影响而发生组织变性坏死。证实斑节对虾烂鳃病由杆状细菌感染引起。本文对细菌引起虾体的损害及对虾的防御机制进行了探讨,认为斑节对虾是由于细菌感染造成鳃呼吸功能障碍和细菌毒血症造成肝胰脏功能障碍而死亡。 相似文献
12.
13.
虾类微量元素含量的主成分分析 总被引：2，自引：0，他引：2
应用主成分分析法对湛江几种养殖对虾和海捕天然对虾的微量元素的含量进行综合评价。结果表明:虾体中微量元素含量均衡;综合排名依次为野生斑节对虾、野生凡纳滨对虾、养殖斑节对虾、养殖凡纳滨对虾,两种野生对虾的综合排名均高于养殖对虾;野生斑节对虾综合得分为1.6708,微量元素含量最均衡,营养价值最高,养殖凡纳滨对虾营养价值最低,综合得分为-1.20943。 相似文献
14.
Hematological Changes m White Spot Syndrome Virus-Infected Shrimp, Fenneropenaeus chinensis (Osbeck)
The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifesta- tions of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and dis- integrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular bemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis. 相似文献
15.
The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV
(white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection,
feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those
in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present
in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM
in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei
often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All
our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed
to be more favorable for WSSV infection in F. chinensis. 相似文献
16.
Peptides in shrimp hemolymph play an important role in the innate immune response.Analysis of hemolymph will help to detect and identify potential novel biomarkers of microbial infection.We used magnetic bead-based purification(ClinProt system) and matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS) to characterize shrimp hemolymph peptides.Shrimp serum and plasma were used as the source of samples for comparative analysis,and it was found that serum was more suitable for shrimp hemolymph peptidomic analysis.To screen potential specific biomarkers in serum of immune-challenged shrimps,we applied magnetic bead-based MALDI-TOF MS to serum samples from 10 immune-challenged and 10 healthy shrimps.The spectra were analyzed using FlexAnalysis 3.0 and ClinProTools 2.1 software.Thirteen peptide peaks significantly different between the two groups were selected as candidate biomarkers of lipopolysaccharide(LPS)-infection.The diagnostic model established by genetic algorithm using five of these peaks was able to discriminate LPS-challenged shrimps from healthy control shrimps with a sensitivity of 90% and a specificity of 100%.Our approach in MALDITOF MS-based peptidomics is a powerful tool for screening bioactive peptides or biomarkers derived from hemolymph,and will help to enable a better understanding of the innate immune response of shrimps. 相似文献
17.
抗菌肽对凡纳滨对虾生长和机体免疫的影响 总被引：1，自引：0，他引：1
基础饲料(对照组)中添加10g/kg抗菌肽制剂喂养体重0.10±0.01g、体长2.17±0.11cm的凡纳滨对虾(Penaeus vannamei)30d,进行10d的水浴攻毒(鳗弧菌10~8/mL),探讨饲料中添加抗菌肽对凡纳滨虾生长、成活率、免疫保护率,以及肌肉中溶菌酶活性、超氧化物歧化酶活性、总抗氧化能力的影响。结果表明:饲料中添加抗菌肽可显著提高凡纳滨对虾相对增重率、成活率和对致病菌的免疫保护率(P<0.05),但对饲料系数无显著影响(P>0.05);未受外界微生物刺激时,饲料中添加抗菌肽对凡纳滨对虾总抗氧化能力有促进作用(P<0.05),受外界微生物刺激后,饲料中添加抗菌肽喂养凡纳滨对虾可提高对虾超氧化物歧化酶活性(P<0.05)和总抗氧化能力(P<0.01)。因此饲料中添加抗菌肽喂养凡纳滨对虾可提高其机体非特异性免疫。 相似文献
18.
WANG Baojie PENG Hongni LIU Mei JIANG Keyong ZHANG Guofan WANG Lei 《中国海洋大学学报(英文版)》2013,12(3):477-483
The clotting protein (CP) plays important and diverse roles in crustaceans, such as coagulation and lipid transportation. A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis (named as Fc-CP) with Q sepharose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography. Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca2+, suggesting that the clotting reaction is catalyzed by a Ca2+-dependent transglutaminase in shrimp hemocytes. The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE. CP exists as disulfide-linked homodimers and oligomers. The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon, Farfantepenaeus paulensis and Litopenaeus vannamei; and similar to that of other decapods. The purified Fc-CP was digested with trypsin and verified on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) mass spectrometry. Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph. 相似文献
19.
模拟吹填、疏浚等海上施工过程所产生的底泥悬浮物浓度的变化规律,探讨不同底泥悬浮物浓度下对凡纳滨对虾仔虾成活影响。结果表明:海水中底泥悬浮物浓度超过10mg/L,持续时间超过12h时,将对凡纳滨对虾成活率产生影响,随着悬浮物实验浓度的递加,仔虾的死亡率从13.33%增至96.67%;超过80mg/L,且持续时间超过48h,将对凡纳滨对虾产生严重影响,甚至导致全部仔虾的死亡;悬浮物对凡纳滨对虾的12h、24h、36h和48h半致死量(LC50)分别为54.56、40.29、25.8和19.18mg/L。 相似文献