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1.
从患出血病罗非鱼的血液、肝、脾分离出嗜水气单胞菌（Ａｅｒｏｍｏｎａｓ ｈｙｄｒｏｐｈｉｌａ）,经动物回归和理化试验鉴定,确诊嗜水气单胞菌为罗非鱼出血病病原;用分离菌制成的饵料吸附型疫苗进行田间试验,免疫组罗非鱼保护率为８７．１％,未免疫组存活率为５９．９％,两组间差异极显著（Ｐ〈０．０１）。 相似文献
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为研究在饲料中添加天然植物复合提取物对罗非鱼感染嗜水气单胞菌后成活率及头肾组织酶活力的影响，试验设计了5个天然植物复合提取物浓度梯度的等氮等能饲料。饲料中天然植物复合提取物的添加量分别为0、0．05％、0．10％、0．15％和0．20％，饲喂8周后，每组取15尾鱼注射嗜水气单胞菌液。试验结果表明，饲料中添加天然植物复合提取物有助于提高感染嗜水气单胞菌后罗非鱼的成活率，并能显著提高头肾溶菌酶、超氧化物歧化酶和酸性磷酸酶的活力。以感染嗜水气单胞菌后的成活率和头肾组织酶活力为指标，罗非鱼饲料中天然植物复合提取物的适宜添加量为0．10％～0．15％。 相似文献
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《中国预防兽医学报》2016,(9)
为评估重庆地区分离的嗜气单胞菌WL3-X2分离株对大鲵的免疫原性,本研究将嗜水气单胞菌WL3-X2分离株经福尔马林灭活,作为免疫原,通过腹腔注射途径间隔2周两次免疫健康大鲵,并检测其体内血清抗体、溶菌酶、补体C3及其免疫保护效应。结果显示,免疫嗜水气单胞菌灭活菌能够显著提高大鲵血清抗体滴度、溶菌酶活性和补体C3含量,首免35 d后这3项指标均达到峰值。此外,对初免4周后的大鲵经肌肉注射0.2 m L浓度为1.0×10~8 cfu/m L剂量的嗜水气单胞菌WL3-X2分离株,进行攻毒保护试验。结果表明,免疫组的相对免疫保护率为66.7%。本研究结果表明,嗜水气单胞菌地方分离株灭活菌对大鲵具有显著免疫保护效力,可以作为大鲵预防嗜水气单胞菌感染的候选疫苗。 相似文献
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为了解嗜水气单胞菌的肉鸭健康带毒、致病与耐药情况,对贵州省三穗县某肉鸭屠宰场临床健康肉鸭随机取样,进行嗜水气单胞菌的分离鉴定、毒力基因检测及耐药性分析。结果显示,分离菌具有嗜水气单胞菌典型的培养特征,菌落形态、菌体形态和生化特性均与嗜水气单胞菌相符;16 S rRNA基因序列系统进化树显示,该分离菌与嗜水气单胞菌聚为一支,同源性均>99%;动物回归试验显示,该分离菌对小鼠有较强的致病性;毒力基因PCR检测显示,该分离菌携带aer、hly、epa、act、alt和ahp等6种毒力基因;药敏试验结果显示,该分离菌对复方新诺明、磺胺嘧啶、环丙沙星和诺氟沙星等15种药物耐药;耐药基因PCR检测显示,该分离菌携带qnrB、Sul1和IntI1等3种耐药基因,与药敏试验表型相符。研究结果为嗜水气单胞菌的生物学特性研究及防控提供参考依据。 相似文献
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致病性嗜水气单胞菌耐喹诺酮类药物菌株的分离鉴定 总被引：4，自引：0，他引：4
对吉林省内12个水域采集和送检的66尾病鱼进行了细菌分离鉴定，共检出46株嗜水气单胞菌疑似菌，通过生化试验结合PCR扩增气溶素基因Aer，确定其中22株为致病性嗜水气单胞菌。进一步采用纸片扩散法和双倍稀释法测定嗜水气单胞菌分离株对多种抗生素的耐药性，其中有4株对喹诺酮类药物耐药，并且为交叉耐药，耐药菌检出率为18．2％，纸片扩散法检测其对喹诺酮类药物的抑菌圈均小于19mm。将病料中分离的8株嗜水气单胞菌负染后电镜观察，耐药菌表面有许多大小不同的球形结构，而敏感菌表面则为细丝状菌毛结构。 相似文献
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疑似草鱼嗜水气单胞菌的分离鉴定及药敏试验 总被引：2，自引：1，他引：1
试验对疑似嗜水气单胞菌草鱼的肝脏、脾脏、肾脏组织进行嗜水气单胞菌的分离鉴定。通过普通营养琼脂、胰蛋白大豆琼脂及生化试剂等的分离培养与生化鉴定后,获得3株细菌;16S rRNA通用引物检测3株细菌的基因序列比对确定菌株后,对确定的草鱼嗜水气单胞菌进行回归试验确认其致病性,然后采用临床上常用的阿奇霉素、阿米卡星、链霉素等10种抗生素进行药敏试验。结果表明,经过分离培养、生化鉴定及16S rRNA通用引物鉴定后确定3种菌株分别为嗜水气单胞菌、嗜麦芽窄食单胞菌和腐败斯瓦尼菌,确定的嗜水气单胞菌回归试验证明其具有致病性;药敏试验显示阿奇霉素对腐败斯瓦尼菌和嗜麦芽窄食单胞菌敏感,其抑菌直径分别为105、95 mm,阿米卡星对嗜麦芽窄食单胞菌、嗜水气单胞菌和腐败斯瓦尼菌这3种菌都较敏感,抑菌直径分别为105、95和93 mm。结果表明,疑似为嗜水气单胞菌病的草鱼有3种细菌感染,其中鉴定的嗜水气单胞菌是致病菌,常用抗生素阿米卡星对3种菌都有抑制作用,而仅对腐败斯瓦尼菌和嗜麦芽窄食单胞菌敏感的是阿奇霉素。 相似文献
7.
应用间接荧光抗体法检测鱼类嗜水气单胞菌的试验研究 总被引：6，自引：0，他引：6
作者制备了鱼类嗜水气单胞菌（AH）－77株兔抗血清,建立了应用间接荧光抗体试验（IFA）检测鱼类嗜水气单胞菌病原的方法。该法能有效地检测人工感染鳟鱼体内的嗜水气单胞菌病原体,对杀鲑气单胞菌、荧光假单胞菌、爱德华氏菌不发生交叉反应。整个过程可在2．5h内完成,是一种有前途的鱼类嗜水气单胞菌快速诊断方法。 相似文献
8.
嗜水气单胞菌是淡水鱼流行性败血症的主要致病菌。在采用添加不同剂量的止痢草精油的饲料投喂3周后,对罗非鱼腹腔注射嗜水气单胞菌,在其攻毒后不同时间取血,通过NBT阳性细胞数量、白细胞数量以及血清溶菌酶活力的变化趋势来研究止痢草精油及病原菌感染对鱼类非特异性免疫功能的相互影响。结果表明,在罗非鱼感染嗜水气单胞菌后72h内,白细胞数量、NBT阳性细胞数量以及血清溶菌酶活力依感染浓度不同,呈现先增加后降低的趋势,而添加了止痢草精油试验组的这3项指标也是先出现增加的趋势,但降低幅度和速度减小,同时到后期恢复到正常水平的速度更快。说明嗜水气单胞菌与罗非鱼非特异性免疫功能之间呈现一定的时间效应,而止痢草精油对罗非鱼的非特异性免疫机能可能有协同增强功效。 相似文献
9.
本试验建立的检测嗜水气单胞菌HEC毒素的点酶法,可在嗜水气单胞菌培养上清和临床病料中检出HEC毒素,可检出的最低水平为95ng/ml。用该法检测大肠杆菌、沙门氏菌、鳗弧菌、荧光假单胞菌、温和气单胞菌等的培养上清及患草鱼出血病的病鱼病料,均为阴性反应,仅与豚鼠气单胞菌、杀鲑气单胞菌新亚种及吕克氏耶尔森氏菌有轻度的交叉反应。对72株病鱼分离菌及33份病鱼组织的检测结果表明,点酶法与溶血试验的符合率分别为80.7%和90.9%。以上试验重复3次,结果完全一致。综上可见,用点酶法检测嗜水气单胞菌HEC毒素,无须作复杂的细菌鉴定,并能直接用于病料,是一个快速、敏感、特异、有效的方法,显示出广阔的应用前景。 相似文献
10.
南京地区池塘水中嗜水气单胞菌的分离鉴定与相关毒力基因检测 总被引：2，自引：0，他引：2
嗜水气单胞菌在自然界广泛存在,是一种典型的人-兽-鱼共患病病原菌。本研究收集南京市多处湖泊池塘水样,采用增菌、平板分离培养、生化试验和种特异性16S rRNA基因扩增等进行嗜水气单胞菌的分离鉴定;采用PCR技术检测分离株毒力基因的分布,进一步通过动物试验确定是否为致病性菌株。结果,从28份水样中共检出嗜水气单胞菌7株,检出率达25%;其中致病性菌株4株,占被检嗜水气单胞菌总数的57.14%(4/7)。本研究有助于了解南京市水体环境中嗜水气单胞菌的污染情况,为进一步预防该菌所引起的相关疾病的发生和传播提供理论依据。 相似文献
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12.
Chu WH Lu CP 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2005,52(4):180-182
The influence of the cytoskeleton on the invasion of Aeromonas hydrophila strain AhJ-1, isolated from diseased fish, in the monolayer cell of epithelioma papillosum cells of carp (EPC) was evaluated by the recovery of gentamicin-resistant bacteria from Triton X-100 cell lysates. The depolymerization of microfilaments (MF) by cytochalasin B and D inhibited the uptake of A. hydrophila in a dose-dependent manner and that of microtubules (MT) by colchicines and nocodazole did not affect the invasion of A. hydrophila in EPC cells significantly. The invasion frequency decreased approximately 62% with the addition of 0.1 microg/ml cytochalasin D and nearly 86% by the addition of 5.0 microg/ml. Invasion decreased approximately 49% and 83% by addition of cytochalasin B in a concentration of 2.5 microg/ml and 10.0 microg/ml. Colchicine and nocodazole, inhibitors of MT formation appears to have little effect on the invasion of EPC cells by strain Ah J-1. Thus MF formation, but not MT formation seems to play an important role in the internalization of A. hydrophila J-1. 相似文献
13.
Protection and serum antibody production against Aeromonas hydrophila was examined in nile tilapia, Tilapia nilotica (L.). Intraperitoneally injected formal in-killed and Freunds complete adjuvant vaccines were compared using different doses (2.9 X 10(7) and 2.9 X 10(9) cfu/ml). Upon challenge, the protective ability and antibody titers resulting were significantly different between vaccinated and unvaccinated groups. A relative level of protection of 100% was obtained within two-weeks, and a maximum level of 53-61% protection was found one-week post-vaccination. 相似文献
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嗜水气单胞菌疫苗的检验及免疫效力试验 总被引：4，自引：1，他引：3
选用嗜水气单胞菌（Ａｅｒｏｍｏｎａｓｈｙｄｒｏｐｈｉｌｉｃ,Ａｈ）疫苗生产用菌Ｊ－１株,以气升式发酵罐２８℃通气培养２８ｈ,制备嗜水气单胞菌疫苗用菌液。半成品检验表明,该疫苗用菌液内无杂菌生长,细菌数量达２．７×１０１０ｃｆｕ／ｍｌ。以０．４％福尔马林溶液３７℃灭活２４ｈ,制成灭活疫苗。经成品检验,该疫苗为棕黄色液体,涂布平板无活菌生长,以１×１０７ｃｆｕ攻击６尾健康鲫鱼,１周内鲫鱼无异常反应。取该疫苗分别以腹腔注射和浸泡两种方式各免疫１２尾鲫鱼,１月后以同源强毒菌株攻击,疫苗的相对存活率分别为８３．３％和５８．３％。 相似文献
16.
Combined effects of beta-glucan and lipopolysaccharide (LPS) on survival and immune response were studied in Cyprinus carpio that were challenged with the pathogen Aeromonas hydrophila. beta-Glucan from Saccharomyces cervisiae and LPS from a virulent strain of A. hydrophila were used in this study. Different concentrations of beta-glucan+LPS mixture were administered on days 1, 7, and 14 through different routes (intraperitoneal injection, bathing, and oral administration). Control and test fish were challenged by intraperitoneal injection of LD50 concentration of A. hydrophila on day 16 and subsequently, mortality and relative percent survival (RPS) were recorded. Intraperitoneal injection elicited 100% RPS even at the lowest concentration (100 microg beta-glucan+10 microg LPS); whereas, oral administration improved RPS rate of carps at higher concentration (1% beta-glucan+0.25% LPS). Bathing did not improve the RPS. Test animals injected with even the minimum dose of the immunomodulators (100 microg beta-glucan+10 microg LPS/fish) had a significant increase in total blood leukocyte counts and an increase in the proportion of neutrophils and monocytes. Superoxide anion production by macrophages was also elevated, which presumably aided the efficient killing of bacterial pathogen. Lower concentration of beta-glucan+LPS had an adjuvant effect on antibody production as pretreatment by injection of 100 microg beta-glucan+10 microg LPS/fish resulted in higher antibody titer against A. hydrophila following vaccination. RT-PCR analyses showed that the expression of interleukin-1beta mRNA did not increase in test fish when compared with the control. Classical and alternative complement pathways were not affected by either the dose or the route of administration of the compounds. It may be concluded that intraperitoneal injection and oral administration, and not the bathing, of beta-glucan+LPS mixture in carp could enhance resistance to challenge by A. hydrophila through changes in several non-specific and specific immune responses. 相似文献
17.
鲤鱼出血病病原体鉴定及其疫苗 总被引：4，自引：0，他引：4
从具有出血症状的鲤鱼肝中分离出一株嗜水气单胞菌,能使实验鲤鱼发生出血症,鉴定为此种疾病的病原,在疾病流行季节中给鲤鱼口服以此株细菌制成的饵料吸附型疫勒后有８８．９％鱼生存,而未服用疫苗的只生存６１．７％。 相似文献
18.
Aeromonas hydrophila is a broad-host-range pathogen and its pathogenesis is multifactorial. A regulatory mechanism known as quorum sensing has been found to be involved in the regulation of virulence in many bacteria. In A. hydrophila the ahyR gene encodes LuxR-type response regulator. Here we describe the inactivation of the ahyR gene of A. hydrophila J-1 by the insertion of a DNA fragment containing a kanamycin resistance determinant and reintroduced by allelic exchange into the chromosome of A. hydrophila J-1 by means of the suicide plasmid pJP5603. Cytotoxic effects on EPC cells assay and LD(50) determinations in fish demonstrated that the ahyR mutant was highly attenuated relative to the wild-type strain. Compared with the parent strain, some characteristics, such as biochemical characters and outer membrane protein profiles, had changed. Some main virulent determinants could not be detected, including proteases, amylase, Dnase, hemolysin and S layer. This article confirmed the important function of AhyR in the pathogenesis of A. hydrophila J-1. 相似文献
19.
根据已发表的嗜水气单胞菌（Aeromonas hydrophila,Ah）溶血素A基因序列设计一对特异性引物,以TPS30菌株基因组为模板,通过PCR扩增溶血素A基因,经T-A克隆、序列测定和分析,结果表明,该基因包含1482 bp碱基,编码494个氨基酸。将溶血素A定向克隆至表达载体pET32a（＋）中,构建重组表达质粒pET-THA,在IPTG诱导下成功获得重组表达蛋白His-GroEL,大小为68 kDa。对该质粒在大肠杆菌BL21（DE3）中的表达特性分析表明,最佳的诱导表达条件为：在0.1 mmol/L的IPTG浓度下,21℃诱导表达3 h。表达的溶血素A蛋白纯化后免疫小鼠,经Western blot检测,表明该蛋白具有免疫原性。对纯化的溶血素A蛋白复性后,进行溶血实验,结果表明该蛋白具有溶血活性。通过本实验的研究,为进一步研究嗜水气单胞菌溶血素的生物学功能奠定了基础。 相似文献