日本鳗鲡腐皮病病原菌的分离及鉴定

从患腐皮病的日本鳗鲡(Anguilla japonica)体表溃烂处分离到1株病原菌(322A),对其进行了人工感染实验和生理生化分析,测定了该菌株的16S rRNA基因序列和促旋酶(gyrase)B亚单位gyrB基因序列,并分别构建系统发育树。结果显示:感染实验证实菌株322A具有致病性;生理生化分析鉴定该菌株属于气单胞菌属(Aero-monassp.);16S rRNA基因分析显示,该菌株与气单胞菌属细菌的同源性均在99%~100%,构建的系统树显示,菌株322A与嗜水气单胞菌(A.hydrophila(FJ462702))亲缘关系最近;gyrB基因分析表明,该菌株与A.hydrophila种内序列的相似性为96%~98%,种间序列的相似性为94%~95%,构建的系统树结果显示,该菌株与A.hydrophila(FJ608553、FJ608552、AF208259)聚为一个分支。综合上述实验结果,菌株322A可鉴定为嗜水气单胞菌(A.hydrophila)。     

环丙沙星对水产动物气单胞菌的药效学研究

环丙沙星为第三代步诺酮类药，是一种安全。高效的抗菌剂。该药对临床分离的几株致病性气单胞菌的最小抑制浓度（MIC）为0.0098μg/ml。人工感染治疗试验表明，该药以1mg/（kg.d）剂量灌服，连用5天，能有效控制鲫鱼败血症的暴发。     

An experimental vaccine against Aeromonas hydrophila can induce protection in rainbow trout,Oncorhynchus mykiss (Walbaum)

A candidate vaccine against Aeromonas hydrophila in rainbow trout, Oncorhynchus mykiss, was developed using a bacterial lysate. To test the strength of protection, A. hydrophila challenge models were compared using injection into both the intraperitoneal (IP) cavity and the dorsal sinus (DS) with selected doses of live bacteria washed in saline or left untreated. Unlike the IP route, injection into the DS with either saline washed or unwashed cells resulted in consistent cumulative mortality and a dose response that could be used to establish a standard challenge having an LD₅₀ of approximately 3 × 10⁷ colony forming units per fish. Survivors of the challenge suffered significantly lower mortality upon re‐challenge than naïve fish, suggesting a high level of acquired resistance was elicited by infection. Passive immunization using serum from hyper‐immunized fish also resulted in significantly reduced mortality indicating protection can be transferred and that some portion of resistance may be antibody mediated. Vaccination of groups of rainbow trout with A. hydrophila lysate resulted in significant protection against a high challenge dose but only when injected along with Freund’s complete adjuvant. At a low challenge dose, mortality in all groups was low, but the bacterial lysate alone appeared to offer some protection.     

华鲮烂尾病病原的分离鉴定及药敏分析

从患有烂尾病的华鲮(Sinilabeo rendahli)体内分离到两个优势菌株(编号:BB090516C-1和BB090516C-2),对分离菌株形态特征、主要理化特性等生物学性状鉴定后测定其16S rRNA序列并构建系统发育树,同时采用K-B琼脂扩散法进行药敏实验。结果显示:经人工感染试验,证实分离菌对华鲮有较强的致病性,该菌为革兰氏阴性、兼性厌氧发酵型短杆菌。经形态特征、培养特性和生理生化反应等指标分析鉴定结果显示该菌为维隆气单胞菌温和生物型(Aerom onas veronii biovar sobria)。用16S rRNA基因序列比对结果显示分离菌株与维隆气单胞菌的同源性高达99%,系统进化树中与维隆气单胞菌自然聚为一支,表明该菌为维隆气单胞菌温和生物型。用22种抗菌类药物进行药敏实验发现,该菌对头孢曲松、硫酸庆大霉素等9种抗菌类药物高度敏感,对新霉素等6种药物中敏,对氨苄青霉素等7种药物耐药。     

鳖源嗜水气单胞菌M-96株表面抗原分析

用ＳＤＳ－ＰＡＧＥ、免疫印迹技术分析嗜水气单胞菌Ｍ－９６株表面抗原，表明该菌体表面抗原是由分子量为３０ＫＤ、３３ＫＤ、３６Ｋ和３９ＫＤ的蛋白质和脂多糖组成，其中部分对胃蛋白酶敏感。     

Comparative pathogenicity of Vibrio spp., Photobacterium damselae ssp. damselae and five isolates of Aeromonas salmonicida ssp. achromogenes in juvenile Atlantic halibut (Hippoglossus hippoglossus)

Juvenile Atlantic halibut (~100 mg, Hippoglossus hippoglossus) were exposed to Vibrio proteolyticus, a Vibrio spp. isolate, Photobacterium damselae ssp. damselae and five different isolates of Aeromonas salmonicida ssp. achromogenes via an hour‐long bath immersion to ascertain their variation in pathogenicity to this fish species. Results were analysed using Kaplan–Meier survival analysis. Analysis of the data from challenges using A. salmonicida ssp. achromogenes revealed three survival values of zero and a spread of values from 0 to 28.43. Challenges using a Vibrio spp isolate, V. proteolyticus and P. damselae resulted in Kaplan–Meier survival estimates of 31.21, 50.41 and 57.21, respectively. As all bacterial species tested could induce juvenile halibut mortalities, they must all be considered as potential pathogens. However, the degree of pathogenicity of A. salmonicida is isolate dependent.     

First isolation of Vibrio tapetis and an atypical strain of Aeromonas salmonicida from skin ulcerations in common dab (Limanda limanda) in the North Sea

Skin ulcerations rank amongst the most prevalent lesions affecting wild common dab (Limanda limanda) with an increase in prevalence of up to 3.5% in the Belgian part of the North Sea. A complex aetiology of these ulcerations is suspected, and many questions remain on the exact factors contributing to these lesions. To construct the aetiological spectrum of skin ulcerations in flatfish, a one‐day monitoring campaign was undertaken in the North Sea. Fifteen fish presented with one or more ulcerations on the pigmented and/or non‐pigmented side. Pathological features revealed various stages of ulcerations with loss of epidermal and dermal tissue, inflammatory infiltrates and degeneration of the myofibers bordering the ulceration, albeit in varying degrees. Upon bacteriological examination, pure cultures of Vibrio tapetis were retrieved in high numbers from five fish and of Aeromonas salmonicida in one fish. The V. tapetis isolates showed cross‐reactivity with the sera against the representative strain of serotype O2 originating form a carpet‐shell clam (Ruditapes descussatus). Moreover, the A. salmonicida isolates displayed a previously undescribed vapA gene sequence (A‐layer type) with possible specificity towards common dab. Further research is necessary to pinpoint the exact role of these agents in the development of skin ulcerations in common dab.     

Exploring the efficacy of vaccine techniques in juvenile sablefish,Anoplopoma fimbria

Wild sablefish, Anoplopoma fimbria, are a valuable commercial species whose populations are declining. Fortunately, sablefish are excellent species for commercial aquaculture. Sablefish raised under high‐density conditions in the marine environment require the use of efficacious vaccines to control disease. Sablefish impacted by disease in net pens may have poor flesh quality and high mortality during grow‐out. As a result, disease can cause financial hardship for sablefish aquaculture operators. The efficacy of a multivalent vaccine preparation for sablefish, administered either by intraperitoneal (i.p.) injection or by immersion, against atypical and typical Aeromonas salmonicida, the causative agents of atypical and typical furunculosis, respectively, was examined. A. salmonicida can affect sablefish at any age and size. Consequently, an efficacious vaccine that can be appropriately and optimally administered to all life stages is desirable. Sablefish vaccinated by immersion at ~1.5 or ~4.5 g with a whole‐cell multivalent vaccine were not protected against either typical or atypical A. salmonicida. Factors that may have contributed to the ineffectiveness of the immersion vaccine are discussed. By contrast, the relative per cent survival (RPS) or potency of the whole‐cell multivalent vaccine injected i.p. in juvenile sablefish at ~50 g against typical and atypical A. salmonicida was 94.3% and 81.7% respectively. The high RPS values indicated that the vaccine successfully initiated an immune response in sablefish upon a second encounter with the pathogen.     

患病蝶尾金鱼维氏气单胞菌温和生物变种的分离鉴定

研究从患病蝶尾金鱼(Butterfly)肝脏中分离获得的1株病菌,为金鱼疾病防治提供参考。分离纯化获得1株细菌,编号为DW-1,通过细菌形态观察、理化特性、16S rDNA序列分析等方法进行鉴定。结果表明DW-1菌株为革兰氏阴性短杆菌,可发酵葡萄糖产气,赖氨酸脱羧酶、硝酸盐还原等为阳性;进一步通过PCR方法扩增16S rDNA序列,测得长度为1 385 bp,与维氏气单胞菌(Aeromonas veronii)相似性为99%~100%;构建系统发育树与维氏气单胞菌温和生物变种(A.veronii biovar sobria)模式菌株ATCC9071聚为一支。最终鉴定该菌株为维氏气单胞菌温和生物变种。人工感染试验结果表明,对斑马鱼半数致死量(LD_(50))为1.17×10~6CFU/m L。药敏试验结果显示该菌株对头孢克肟、头孢哌酮、链霉素、恩诺沙星、诺氟沙星、左氟沙星、复方新诺明等23种药物敏感。     

水族箱气单胞菌的鉴定及致病特性分析

为了确定南京市某渔场发病鱼感染的病原,本研究采集患鱼脏器,采用平板培养、生化实验和特异性gyrB基因扩增测序等方法进行细菌的分离鉴定;利用PCR技术检测分离株毒力基因的分布,分析其生物学特性,并进一步通过动物实验确定菌株致病力.结果分离到1株水族箱气单胞菌,命名为LK-25.该菌株携带5种主要毒力基因:气溶素(aer)、细胞毒性肠毒素(act)、细胞兴奋性肠毒素(alt)、温敏胞外蛋白酶(epr)和丝氨酸蛋白酶(ahp),其溶血性和溶蛋白能力较强,对斑马鱼的半数致死量为1.02×103 CFU/尾,确定为强毒株.进化树分析表明,水族箱气单胞菌与嗜水气单胞菌达卡亚种亲缘关系较近.本研究在国内首次报道发现水族箱气单胞菌,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基础.