鳖病防治技术问答(中)

<正> 9.问:红脖子病和大脖子病是不是一种病? 答:红脖子病和大脖子病是一种病。病鳖背甲失去光泽而呈暗黑色,颈部肿大,行动迟缓,鳖体消瘦。随着病情的日趋严重,颈部充血发红,成为红脖子现象,以致颈部不能正常伸缩。严重时,病鳖眼睛失明,口鼻、舌头都出血。解剖病鳖,见肝、脾脏均肿大,质脆易碎,口腔、食道、胃、肠的粘膜层呈明显的点状、弥蔓性出血占80％。从红脖子病鳖分离到病原菌是嗜水气单胞菌嗜水亚种(Aeromonas,hydrophila sub-sp.hydrophila);从大脖子病鳖分离到的也是嗜水气单

淡水养殖鱼类暴发性传染病致病细菌的研究

于1990—1991年的5—10月间,从湖北、湖南、河南和广东4省的9种淡水养殖鱼类的发病鱼睥、肝、肾中分离的93株细菌为材料,经100多个表型特征和细菌DNA中G+Cmol％遗传型特性测定等研究结果,可归属为产气单胞菌属中的运动性嗜中温产气单胞菌和弧菌属中的河弧菌。按照国际细菌命名法规优先律应为点状产气单胞菌[Aeromonas punctata(Zimmermann)nom.rev];而河弧菌与以前所描述的生物变种Ⅰ、Ⅱ型不同:在43℃能生长;发酵水杨苷产酸;利用D-葡糖酸钙、腐胺;不能利用戊二酸,D

鲢鳙鱼流行性传染病的流行病学和细菌病因的初步研究

1990年3～10月,作者对湖北、湖南,河南三省部分地区的28个养殖场进行了鲢鳙鱼流行病学调查和致病菌的初步研究。结果显示:在调查范围内发病率达60%以上;受感染的除鲢、鳙外,还有鲫、鳊(鲂)、鲤、草鱼以及其它野杂鱼、河蚌;分离到138株细菌,经鉴定可归为三类,早期(3～4月)出现的鲁克氏耶尔森菌(Yersinia ruckeri)、中后期(5月下旬以后)出现的产气单胞菌(Aeromonas sp.)及弧菌(Vibrio sp.)。结合水质环境条件分析,致病细菌是继发侵入者,原发病因除水体的理化因子外,不

鳗鲡烂尾病病原菌的研究

鳗鲡烂尾病是鳗鲡养殖中的一种常见病,主要流行于夏季。在日本报道了由枉状屈桡杆菌引起的鳗鲡烂尾病。本文记述了用TYE培养基从广东潮安养鳗场病鳗中分离到的另外一种引起烂尾病的病原菌,并研究了该病原菌的致病作用,生物学和生理生化特性,鉴定为点状产气单胞杆菌(Aeromonas punctata)。该菌对青霉素、新生素、磺胺噻唑不敏感,而对呋唑唑酮、土霉素、氯霉素、合霉素和金霉素等敏感。

塔式生物滤池处理洗涤剂工业废水的模拟试验

本项研究工作表明,塔式生物滤池在处理模拟洗涤剂工业废水时,能够适应和克服一般好氧生化法所不能解决的泡沫问题,并对废水中的LAS和COD具有一定的去除效果。根据实验结果,初步认为塔滤可应用于洗涤剂工业废水的生化处理,并向洗涤剂行业首次推荐这种废水生物学净化方法。从塔滤的生物膜中分离出了优势菌,经鉴定为一种气单胞菌(Aeromonas sp.D-4)。

二龄草鱼肠炎病发病机理

本文通过对患肠炎病的二龄草鱼病鱼和正常草鱼的肠道及血液中的产气单胞菌(Aeromonas)的数量和毒力的比较,病鱼肠道、血液和其他内脏中的产气单胞菌所出现比率的调查,不同水温对该菌致病力的影响和用菌体提取液接种鱼体,以及用鲎试剂(鲎变形细胞溶解物Tachypleus Amebocyte Lysatc “TAL”)对病鱼和正常鱼体内的内毒素进行测定等一系列的试验和观察,从而推断出二龄草鱼细菌性肠炎病的发病机理。

气单胞菌(Aeromonassp.D-4)降解LAS的研究

Aeromonas sp.D-4不能以LAS作为唯一碳源,它对LAS的利用是通过共代谢来完成的。LAS对D-4具有毒害作用,而且,起始LAS浓度越高,毒害作用越大。LAS的最大去除率与起始LAS浓度呈负相关。当起始LAS在40—120mg/L之间时,去除率较高;如果起始LAS在40mg/L左右,则去除率可达80％以上。研究还表明,Aeromonas sp.D-4纯培养对LAS的去除率(最大值84.97％)大于混合菌(最大值78.57％)。耗氧呼吸测定证实了Aeromonas sp.D-4对于LAS的共代谢

斑点气单胞菌对草鱼的毒力

<正> 斑点气单胞菌(Aeromonas punctata)是危害草鱼(Ctenopharyngodon idellus Cuv. et val.)的主要病原菌之一同时也危害鲢、鳙等多种鱼类。但作为一种病原菌对鱼类的毒力有多大,乞今还没有可靠的报道。

三角帆蚌死亡情况的调查研究

<正> 当前淡水珍珠生产大多是采用三角帆蚌育珠。作为育珠生产用的三角帆蚌,在生产过程中常大批死亡,造成重大经济损失,为广大蚌珠生产者及有关科研人员所关注。作者曾在1982年报道了三角帆蚌细菌性疾病的研究,认为点状产气单胞菌的一新亚种—帆蚌点状产气单胞菌(Aeromonas punctata

Immunoglobulin joining (J) chain and its expression in the Chinese soft-shelled turtle (Pelodiscus sinensis)

The immunoglobulin (Ig) joining (J) chain plays an important role in the formation of polymeric Igs and their transport into secretions. In the present study, the cDNA sequence of J chain has been cloned from the Chinese soft-shelled turtle (Pelodiscus sinensis) by reverse transcription (RT)-PCR and rapid amplification of cDNA ends (RACE). The cDNA sequence is 2347 bp in length and contains an open reading frame of 480 bp encoding 160 aa including the signal sequence. The deduced amino acid sequence has a high degree of homology with that of an already reported turtle J chain (80.7%), and of chicken (71.3%). By using real-time quantitative RT-PCR analysis, a significant up-regulation of J-chain transcripts was observed in spleen, kidney and blood of turtles injected with inactivated Aeromonas hydrophila, indicating the immune role of J chain in response to bacterial infection. (C) 2009 Elsevier B.V. All rights reserved.

IgM, IgD and IgY and their expression pattern in the Chinese soft-shelled turtle Pelodiscus sinensis

Three Ig isotypes, IgM, IgD, and IgA, were previously known in reptiles. Here, in this report we describe IgM, IgD and a novel immunoglobulin heavy-chain isotype upsilon (IgY) in Chinese soft-shelled turtle (Pelodiscus sinensis). The IgM and IgY constant domains are characteristically similar to their counterparts described in other vertebrates. The expression of IgM and IgD were detected at mRNA level early during embryonic development, and their expression increased during further development. However, the IgY expression was not detected in larval turtles until 90 days after hatching-out. The increase in the transcription of these three Ig molecules was analyzed by using real-time PCR in spleen, kidney and blood following the injection of inactivated Aeromonas hydrophila. The primary increase in the expression of these three Igs was observed I week after the first injection, although not statistically significant, and the second injection 2 weeks after the first injection provoked a significant increase in the expression of these Igs, revealing a pattern of primary and secondary antibody response in the turtle. The present study represents the first report on reptile IgY and the pattern of IgM, IgD and IgY transcription in reptiles. (C) 2009 Elsevier Ltd. All rights reserved.

Molecular characterization and expression profiles in response to bacterial infection of Chinese soft-shelled turtle interleukin-8 (IL-8), the first reptilian chemokine gene

In this study, an IL-8 homologue has been cloned and identified from a reptile, Chinese soft-shelled turtle for the first time. The full-length cDNA of turtle IL-8 was 1188 bp and contained a 312 bp open reading frame (ORF) coding for a protein of 104 amino acids. The chemokine CXC domain, which contained Glu-Leu-Arg (ELR) motif and four cysteine residues, was well conserved in turtle IL-8. The 4924 bp genomic DNA of turtle IL-8 contained four exons and three introns. Phylogenetic analysis showed that the amino acid sequence of turtle IL-8 clustered together with birds. RT-PCR analysis showed that turtle IL-8 mRNA was constitutively expressed liver, spleen, kidney, heart, blood and intestine tissues of control turtles. Real-time quantitative PCR analysis further indicated that the turtle IL-8 mRNA expression was apparent in various tissues at 8 h and up-regulated significantly during 8 h-7 d after Aeromonas hydrophila infection. The present studies will help us to understand the evolution of IL-8 molecule and the inflammatory response mechanism in reptiles. (C) 2009 Elsevier Ltd. All rights reserved.

C1q-like inhibits p53-mediated apoptosis and controls normal hernatopoiesis during zebrafish embryogenesis

Except for the complement C1q, the immunological functions of other C1q family members have remained unclear. Here we describe zebrafish C1q-like, whose transcription and translation display a uniform distribution in early embryos, and are restricted to mid-hind brain and eye in later embryos. In vitro studies showed that C1q-like could inhibit the apoptosis induced by ActD and CHX in EPC cells, through repressing caspase 3/9 activities. Moreover, its physiological roles were studied by morpholino-mediated knockdown in zebrafish embryogenesis. In comparison with control embryos, the C1q-like knockdown embryos display obvious defects in the head and cramofacial development mediated through p53-induced apoptosis, which was confirmed by the in vitro transcribed C1q-like mRNA or p53 MO co-injection. TUNEL assays revealed extensive cell death, and caspase 3/9 activity measurement also revealed about two folds increase in C1q-like morphant embryos, which was inhibited by p53 MO co-injection. Real-time quantitative PCR showed the up-regulation expression of several apoptosis regulators such as p53, mdm2, p21, Box and caspase 3, and down-regulation expression of hbae1 in the C1q-like morphant embryos. Knockdown of C1q-like in zebrafish embryos decreased hemoglobin production and impaired the organization of mesencephalic vein and other brain blood vessels. Interestingly, exposure of zebrafish embryos to UV resulted in an increase in mRNA expression of C1q-like, whereas over-expression of C1q-like was not enough resist to the damage. Furthermore, C1q-like transcription was up-regulated in response to pathogen Aeromonas hydrophila, and embryo survival significantly decreased in the C1q-like morphants after exposure to the bacteria. The data suggested that C1q-like might play an antiapoptotic and protective role in inhibiting p53-dependent and caspase 3/9-mediated apoptosis during embryogenesis, especially in the brain development, and C1q-like should be a novel regulator of cell survival during zebrafish embryogenesis. (c) 2008 Elsevier Inc. All rights reserved.

Molecular cloning, characterization and expression analysis of the PKZ gene in rare minnow Gobiocypris rarus

Double-stranded RNA-activated protein kinase (PKR) plays an important rote in interferon-induced antiviral responses, and is also involved in intracellular signaling pathways, including the apoptosis, proliferation, and transcription pathways. In the present study, a PKR-like gene was cloned and characterized from rare minnow Gobiocypris rarus. The full length of the rare minnow PKR-like (GrPKZ) cDNA is 1946 bp in Length and encodes a polypeptide of 503 amino acids with an estimated molecular mass of 57,355 Da and a predicted isoelectric point of 5.83. Analysis of the deduced amino acid sequence indicated that the mature peptide contains two Zalpha domains and one S_TKc domain, and is most similar to the crucian carp (Carassius auratus) PKR-like amino acid sequence with an identity of 77%. Quantitative RT-PCR analysis showed that GrPKZ mRNA expression is at low levels in gill, heart, intestine, kidney, liver, muscle and spleen tissues in healthy animals and up-regulated by viruses and bacteria. After being infected by grass carp reovirus, GrPKZ expression was up-regulated from 24 h post-injection and lasted until the fish became moribund (P < 0.05). Following infection with Aeromonas hydrophila, GrPKZ transcripts were induced at 24 h post-injection (P < 0.05) and returned to control levels at 120 h post-injection. These data imply that GrPKZ is involved in antiviral defense and Toll-like receptor 4 signaling pathway in bacterial infection. (C) 2008 Elsevier Ltd. All rights reserved.

Gene structure of an antimicrobial peptide from mandarin fish, Siniperca chuatsi (Basilewsky), suggests that moronecidins and pleurocidins belong in one family: the piscidins

The gene of piscidin, an antimicrobial peptide, has been cloned from the mandarin fish, Siniperca chuatsi. From the first transcription initiation site, the mandarin fish piscidin gene extends 1693 nucleotides to the end of the 3' untranslated region and contains four exons and three introns. A predicted 79-residue prepropeptide consists of three domains: a signal peptide (22 aa), a mature peptide (22 aa) and a C-terminal prodomain (35 aa). The shortage of XQQ motif in the prodomain of mandarin fish piscidin and the similar gene structure between moronecidins (piscidins) and pleurocidins may indicate that they are derived from the same ancestor gene. We thus suggest that piscidin should be used as a terminology for these antimicrobial peptides in the future. The mandarin fish piscidin mRNA was abundant in intestine, spleen, pronephros and kidney analysed by real-time polymerase chain reaction. After stimulation with lipopoly saccharides (LPS), a marked increase in transcripts was observed in most tissues, indicating that piscidin is not only a constitutively expressed molecule, but also has an increased response to bacterial infection. The synthetic, amidated mandarin fish piscidin exhibited different antimicrobial activity against different fish bacterial pathogens, especially against species of Aeromonas, which may to certain extent reflect the pathogenicity of these bacteria.