斑点叉尾■溃烂症的病原鉴定和致病特性

为探明斑点叉尾■(Ictalunes punctatus)溃烂症的病因,从4尾患鱼肝脾中分离纯化出4株优势菌株,并进行病原鉴定、毒力基因检测、动物回归感染和药敏试验。4株优势菌经鉴定并命名为杀鲑气单胞菌无色亚种(Aeromonas salmonicida subsp achromogenes)X-G1,杀鲑气单胞菌杀鲑亚种(A.s subsp salmonicida)X-P2、X-P3和嗜水气单胞菌(A.hydrophila)X-P4。15℃时,杀鲑气单胞菌X-G1、X-P2和X-P3的世代时间(约14 min)均小于嗜水气单胞菌X-P4(约20 min);25℃时,杀鲑气单胞菌X-G1、X-P2和X-P3株的世代时间(约20 min)均大于嗜水气单胞菌X-P4株(约16 min)。X-G1株可检到弹性蛋白酶、溶血素和甘油磷脂胆固醇酰基转移酶等3种毒力基因;X-P2株仅可检到弹性蛋白酶1种毒力基因;X-P3株可检测到弹性蛋白酶、溶血素、细胞毒性肠毒素、丝氨酸蛋白酶、酯酶、气溶素和甘油磷脂胆固醇酰基转移酶等7种毒力基因;X-P4株可检测到鞭毛、弹性蛋白酶、气溶素、细胞毒性肠毒素、热不稳定...     

一株感染深水网箱养殖许氏平鲉的病原菌分离与鉴定

2016年7月,山东省长岛县深水网箱养殖许氏平鲉(Sebastes schlegeli)暴发严重皮肤溃疡症。作者对病鱼进行病原菌分离。通过形态学观察、常规生理生化试验、gyrB和16S rDNA基因克隆测序等方法对分离菌株进行鉴定。在病灶溃疡处分离到一株绝对优势菌BZ01,该菌株在TSB固体培养基上呈半透明菌落,在TCBS选择性培养基上菌落呈绿色。透射电镜观察为短棒状,具有单根极生鞭毛。人工回接感染证明,该菌株对许氏平鲉具有较强的致病力,可以引起皮肤溃疡等症状,且与自然感染症状一致,其LD₅₀为2.07×10~6 CFU/ml。通过gyrB和16S rDNA基因序列测定并构建系统发育树显示,菌株BZ01与弧菌属同源性最高,并在系统发育树中与轮虫弧菌(Vibrio rotiferianus)聚为一枝,结合形态及生理生化表型测定结果,将该菌株鉴定为轮虫弧菌(V.rotiferianus)。药敏试验结果显示,该菌株对四环素类、喹诺酮类、香豆素类、肽酰转移酶类高度敏感,而对大环内酯类、多肽类、磺胺类、β-内酰胺类、氨基糖苷类中度敏感或不敏感。     

Biological characteristics and pathogenicity of a highly pathogenic Shewanella marisflavi infecting sea cucumber, Apostichopus japonicus

Shewanella marisflavi isolate AP629 is described as a novel pathogen of sea cucumber. The LD(50) values (14 days) in sea cucumber, mice and swordtail fish were 3.89 × 10(6) , 6.80 × 10(4) and 4.85 × 10(4) CFU g(-1) body weight, respectively. Studies on S. marisflavi were conducted, including morphology, physiological and biochemical characteristics, haemolysis, whole-cell protein and 16S rDNA gene sequence. Colonies of S. marisflavi appeared faint red on marine agar and green on thiosulphate-citrate-bile salt-sucrose media. Shewanella marisflavi had polar flagella. The cells were Gram-negative, oxidase- and catalase-positive and not sensitive to O/129. The bacterium exhibited β-haemolysis on sheep blood agar and produced H(2) S. Shewanella marisflavi survived and grew at 4-35°C, pH 6.0-9.2 and in the presence of 0-8% NaCl. The whole-cell proteins included 13 discrete bands, and proteins of molecular weight 87, 44 and 39 kDa were found in all five strains of Shewanella spp. The difference in 16S rDNA gene sequences in S. marisflavi was at the 446 bp site: S. marisflavi (KCCM 41822) - G, isolate AP629 - A. This is the first report that Shewanella is pathogenic to sea cucumber.     

Analysis of exotoxins produced by atypical isolates of Aeromonas salmonicida,by enzymatic and serological methods

In this study, exotoxins produced by 62 Aeromonas salmonicida strains and the bacterium Haemophilus piscium were analysed. Enzymatic assays, zymograms and serological detection were used to monitor secretion by bacterial strains of the previously described exotoxins P1, GCAT and AsaP1 and also the extracellular P2 metallo-gelatinase and a serine caseinase, which is different from the P1 protease and has not yet been characterized. Based on the results, the strains were divided into five groups. One comprised the type strains for A. salmonicida ssp. masoucida, H. piscium and 36% of the atypical isolates, and another, a type strain for A. salmonicida ssp. smithia together with 14% of the atypical isolates. A second type strain of A. salmonicida ssp. smithia was grouped with 8% of the atypical isolates. The largest group contained the type strains for A. salmonicida ssp. achromogenes and 38% of the atypical isolates. The type strains for A. salmonicida ssp. salmonicida were in the last group with all the four typical strains and 4% of the atypical isolates. The combination of zymogram and serological detection used is recommended as the most reliable method for characterizing A. salmonicida strains according to their exotoxin secretion.     

人工鱼礁区不同网目尺寸单片刺网对许氏平鲉和大泷六线鱼的选择性

为探究人工鱼礁区单片刺网对许氏平鲉(Sebastes schlegeli)和大泷六线鱼(Hexagrammos otakii)的选择性, 于 2021 年 10 月在荣成北部人工鱼礁区使用 4 种网目尺寸(40 mm、50 mm、60 mm、70 mm)的单片刺网进行选择性实验。在 SELECT 模型的架构下, 使用 Normal、Lognormal、Gamma、Bi-normal 模型进行选择性拟合, 根据赤池信息指数(AIC)和模型残差(MD)选择最佳拟合模型。结果表明: Lognormal 模型拟合效果最佳, 其 AIC 值和 MD 值均为最低值。根据 Lognormal 模型, 不同网目尺寸许氏平鲉的模型拟合长度分别为 114.23 mm、142.79 mm、 171.35 mm、199.90 mm; 大泷六线鱼的模型拟合长度分别为 129.67 mm、162.09 mm、194.51 mm、226.92 mm。许氏平鲉和大泷六线鱼的体长与最大剖面周长的线性关系分别为: G=0.2877L+0.7463(R² =0.6722), G=0.2468L– 3.8531(R² =0.6851)。选取 150 mm 作为许氏平鲉和大泷六线鱼的理论成熟体长(MBL)进行分析, 结合幼鱼留存比例, 发现网目尺寸为 60 mm 和 70 mm 的刺网能够保护人工鱼礁区许氏平鲉和大泷六线鱼的幼鱼资源。研究结果可为人工鱼礁区渔业资源的管理和养护提供参考。     

Evaluation of profiles of Aeromonas salmonicida as epidemiological markers of furunculosis infections in fish

Abstract. Strains of Aeromonas salmonicida subsp. salmonicida , the agent of furunculosis disease of salmonid fish, have fairly uniform plasmid patterns. Of 35 strains examined by agarose gel electrophoresis, 28 had a pattern consisting of four small plasmids (4.2, 3.6, 3.5, 3.3 Mda) and a larger plasmid. The larger plasmid was most often 50–56 Mda, but it was larger in some strains. In the remaining seven strains, the same general profile was seen, but one of the small plasmids was missing. An additional plasmid was present in six strains. The pattern seen in 30 strains collected from Ontario fish over an 8-year period did not differ significantly from five reference isolates from other locations. Plasmid profiles of A. salmonicida strains appear too uniform to provide a useful epidemiological tool. The non-pigmented. atypical strains of A. salmonicida subsp. achromogenes and A. salmonicida subsp. masoucida, A. media , and brown-pigmented strains of A. hydrophila had different plasmid DNA profiles, which were distinct from those of typical isolates of A. salmonicida subsp. salmonicida . Antibiotic susceptibility patterns, determined by the agar dilution method, were uniform for most typical strains. A non-transferable resistance to tetracyclmes was found in two Ontario isolates, but antibiotic resistance was relatively uncommon among the Ontario isolates.     

Clinical, microbiological and epidemiological findings in recent outbreaks of goldfish ulcer disease due to atypical Aeromonas salmonicida in south-eastern Australia

Abstract. The spread of goldfish ulcer disease (GUD) from Victoria to New South Wales, Australia, and the first isolation of Aeromonas salmonicida from wild goldfish are reported. Cultural, biochemical and protein SDS-PAGE characteristics of these recent isolates are compared with those of existing Australian isolates, with strains recovered from goldfish in Italy and the USA (atypical strains) and with strain ATCC 14174 (typical strain). The Australian isolates were identical and closely resembled the exotic atypical strains. Although there were several biochemical differences between the atypical isolates and the typical ATCC 14174 strain, the results of SDS-PAGE confirmed that these strains were closely related. The homology of the Australian and overseas strains recovered from goldfish supports the common view that A. salmonicida was introduced first into Australia with diseased goldfish in 1974. The three widely separated outbreaks of GUD reported here confirm that an atypical strain of A. salmonicida is now endemic in Australia.     

黑龙江地区鲤春病毒血症病毒的分离与基因型分析

对2015—2016年黑龙江不同地区的40个养殖场送检的鲤(Cyprinus carpio)进行鲤春病毒血症病毒(spring viremia of carp virus,SVCV)的细胞培养分离、PCR鉴定、病毒滴度测定、病毒表面糖蛋白(glycoprotein,G)氨基酸序列聚类分析及基因分型研究。细胞培养结果显示,来自4个不同养殖场的鲤组织样本能够感染鲤上皮细胞(epithelioma papulosum cyprini,EPC)产生典型细胞病变(cytopathic effect,CPE),收集病毒悬液分别称为Shlj1~Shlj4。PCR鉴定结果表明,该4株病毒均为SVCV。病毒滴度实验测算出SVCV Shlj 1~Shlj 4的滴度分别为10~(6.28)、10~(6.88)、10~(7.57)和106.38 TCID50/mL。Shlj的糖蛋白基因核苷酸序列的聚类分析和遗传进化分析结果显示,Shlj 1~Shlj 4与Gen Bank收录的中国参考株A2、BJ0505-2和美国参考株USA、212364聚为一簇,同源性为98.4%~99.8%;Shlj 1~Shlj 4毒株之间的糖蛋白核苷酸序列相似性在98.6%~99.8%,其中Shlj 3与美国SCVC毒株USA、212364具有最高的核苷酸相似性(99.8%),Shlj 2与英国参考毒株880163具有最低的相似性(88.0%)。糖蛋白氨基酸序列比对结果显示,Shlj4中氨基酸突变最多,与另3个毒株差异较大。基因型分析结果显示,Shlj 1~Shlj 4均为基因Ia型。本研究结果表明,黑龙江地区2015—2016年间的SVCV检出率约为10%,并且来源于不同养殖场的病毒分离株的核酸序列呈现不同程度的差异,该结果进一步证明SVCV毒株在中国不同的鲤养殖环境中正在不断地进化。     

Recovery potential of black rockfish,Sebastes melanops Girard,recompressed following barotrauma

Overfished species of rockfish, Sebastes spp., from the Northeast Pacific experience high bycatch mortality because of ‘barotrauma’, a condition induced from the rapid change in pressure during capture. Field experiments show that it may be possible for rockfish to recover from barotrauma if quickly recompressed; however, no work has followed the physiological recovery of rockfish after recompression or determined whether it is possible for rockfish to survive barotrauma in the long term. Barotrauma was induced in adult black rockfish, Sebastes melanops Girard, from a simulated depth of 35 m, followed by recompression. Blood and selected tissues (eye, heart ventricle, head kidney, liver, rete mirabile and gonad) were sampled at days 3, 15 and 31 post‐recompression to evaluate the tissue‐ and physiologic‐level response during recovery. No mortality from barotrauma occurred during the experiments, and feeding resumed in 80% of both treatment and control fish. The primary injury in treatment fish was the presence of a ruptured swimbladder and/or a ruptured tunica externa (outer layer of swimbladder), which was slow to heal. Blood plasma was analysed for glucose, sodium, chloride, potassium, calcium, phosphorus, insulin‐like growth factor‐1 and cortisol. Plasma analyses indicated no strong effects because of barotrauma, suggesting overall handling stress outweighed any effect from barotrauma. Rockfish with ruptured swimbladders may face compromised competency in the wild; however, it appears the majority of black rockfish decompressed from 35 m have a high potential for recovery if recompressed immediately after capture. This research suggests recompression could be a valuable bycatch mortality reduction tool for rockfish in recreational fisheries.     

Non-pigment-producing isolates of Aeromonas salmonicida subspecies salmonicida: isolation, identification, transmission and pathogenicity in Atlantic salmon, Salmo salar L.

Four non-pigment-producing isolates and two pigment-producing isolates of Aeromonas salmonicida sp. salmonicida were isolated from the head-kidney of diseased farmed Atlantic salmon, Salmo salar L. The cultural, morphological and biochemical features of the isolates were compared with those of reference strains. Injection and cohabitation experiments were performed. The only difference between the non-pigment-producing isolates and the pigment producing reference strains of A. salmonicida ssp. salmonicida was the inability of the former to produce pigment. In the injection experiments, the investigated non-pigment-producing isolate produced a significantly higher mortality compared with the mortality caused by the reference strain, whereas no difference in mortality was detected in the cohabitation experiments.     

Availability of squid viscera meal with cadmium removal treatment as a feed ingredient for fingerling black rockfish Sebastes schlegeli

Two 12-week feeding experiments were conducted to evaluate the nutritional value of squid viscera meal with cadmium removal treatment (dCSVM), which contained 1.5–2.0 mg/kg cadmium, as an alternative protein source to sardine meal in diets for fingerling black rockfish Sebastes schlegeli. Initial mean body weights in experiments 1 and 2 were 23.0 and 6.4 g, and the replacement rates of sardine meal with dCSVM in the test diets were 20–80 % and 30–60 %, respectively. In experiment 1, inclusion of dCSVM at 40 % and higher replacement rates of sardine meal retarded the growth of fish due partly to inferior dCSVM protein digestibility. The cadmium (Cd) concentrations in fish muscle of all treatment groups were below the detection limit (<0.1 mg/dry matter). In experiment 2 using dCSVM containing Cd at a lower level and having a higher protein digestibility than in experiment 1, no significant differences were observed in the growth between the control and 60 % replacement groups, although feed efficiency gradually decreased with the increase of dCSVM inclusion. These results show that dCSVM is safe and useful as an alternative protein source and could replace up to 60 % of sardine meal in fingerling black rockfish diets.     

Cloning and characterization of the haemolysin determinants from Aeromonas hydrophila

Abstract. Two haemolysin genes (AHH4 and AHH-2) of Aeromonas hydrophila ATCC7966 were cloned into a plasmid vector in Escherichia coli K-12. An open reading frame (ORF) of the AHH-1 haemolysin gene was 1734 base pairs (bp). and corresponded to a protein of 577 amino acid residues. Analysis of the deduced amino sequence indicated a highly hydrophobic N-terminal region which had the characteristics of a leader peptide. The sequence also included the -10 region and the -35 region of a promoter, and a ribosome- binding site upstream from the ORF. The termination site was located downstream from the ORF. The haemolysin was a thermolabile protein with the predicted molecular mass of 60 kDa. The AHH-1 gene is distributed in various A. hydrophila and A. salmonicida strains. The nucleotide sequence of a 981 bp ORF of the AHH-2 gene was encoded with the predicted molecular mass of 377 kDa polypeptides. The homology of the nucleotide sequence was very low between the AHH-1 and AHH-2 genes, and also with the aerolysin gene cloned by Howard & Buckley (19S6). No leader peptide was found in the N-terminal region of the ORF of the AHH2 gene. The AHH-2 gene was detected in the original strain ATCC7966, but was not detected in other tested strains of A. hydrophila and A. salmonicida.     

洪泽湖河蚬(Corbicula fluminea) 2种表型群体的遗传变异分析

采用形态学分析(壳长、壳宽和壳高)和分子标记技术(细胞色素氧化酶Ⅰ基因,COⅠ)对洪泽湖河蚬(Corbicula fluminea)黄色和黑色2个群体的形态和遗传多样性特征进行研究.形态参数统计分析结果显示,2个群体的形态特征之间有显著性差异.经PCR扩增和序列测定,获得614 bp COⅠ基因序列,2个群体的COⅠ基因序列的碱基组成高度一致,均表现出A+T的含量(64.8％)明显高于G+C的含量(35.2％).28个黑色个体发现7种单倍型,单倍型多样性和核苷酸多样性分别为0.794和0.04274;30个黄色个体发现5种单倍型,单倍型多样性和核苷酸多样性分别为0.607和0.02825.单倍型之间的遗传距离在0.002-0.091之间,其NJ和MP系统发生树表明,COⅠ基因单倍型聚为2个明显分支.分子方差分析(AMOVA)结果显示,Fst=0.21736 (P＜0.01),21.74％的变异来自群体间,78.26％的变异来自群体内,2个群体之间有显著的遗传分化.研究表明,应将洪泽湖河蚬黑色和黄色群体分别作为独立单元进行管理和保护.     

In vitro activities of oxolonic acid, ciprofloxacin and norfloxacin against Aeromonas salmonicida

Abstract. Oxolinic acid and two new fluoroquinolones, ciprofloxacin and norfloxacin, were evaluated for in vitro antibacterial activity against Aeromonas salmonicida. Although oxolinic acid was as active as ciprofloxacin in terms of minimum inhibitory concentration (MIC), fluoroquinolones were significantly more active in terms of their ability to kill both oxolinic acid sensitive and resistant strains of A. salmonicida. Furthermore, the fluorinated drugs were active against non-dividing A. salmonicida. It would appear worthwhile to carry out further investigations with fluoroquinolones as they may be more effective in treating A. salmonicida infections than the current regime of oxolinic acid.     

Comparison of oxygen consumption patterns between wild and cultured black rockfish Sebastes schlegeli

ABSTRACT:   To compare metabolic activity rhythms between wild and cultured black rockfish Sebastes schlegeli , we measured long-term oxygen consumption rates (OCR) using an automatic intermittent-flow respirometer under constant temperature and darkness. Oxygen consumption rates peaked among wild black rockfish at 12.4 h intervals, which corresponded to a circatidal rhythm. The wild fish were probably exhibiting responses that corresponded to tidal events in their natural environment. However, when captured wild black rockfish were kept under laboratory conditions (12 h light [L] : 12 h dark [D]) for 30 days, the OCR shifted to a circadian rhythm (24.1–24.9 h). The OCR of cultured black rockfish that had been reared in a tank for 9 months peaked at approximately 24 h intervals, corresponding to a circadian rhythm. The results of this study suggest that the differences in OCR patterns between wild and cultured fish were mainly due to differences in the environmental conditions between tidal and non-tidal habitats.     

Aeromonas salmonicida infection in wrasse (Labridae), used as cleaner fish, on an Atlantic salmon, Salmo salar L., farm

Abstract. Typical Aeromonas salmonicida with similar biochemical characteristics to A. salmonicida from Atlantic salmon, was isolated from wrasse, Ctenolabrus rupestris (L.) and Centrolabrus exoletus (L.), stocked as cleaner fish with these salmon. Although no external clinical signs were apparent, localized bacterial microcolonies were observed in muscle, gills, intestine, kidney and myocardial tissue. Mortalities attributed to A. salmonicida comprised 55% (n = 32) of total mortalities. No carriers of A. salmonicida were found in wild wrasse following stress testing. Although salmon post-smolts died when challenged with 1 × 10⁵ ml^-1 of a virulent strain, there were no mortalities in challenged wrasse. An oral route of infection is suggested rather than water-borne transfer as wrasse browsed on salmon mortalities. Wrasse were treated for A. salmonicida infection by injection with antibiotic and were also vaccinated, and in the latter case, elevation of antibody levels was noted.     

养殖裸盖鱼(Anoplopoma fimbria)疥疮病病原菌的分离与鉴定

裸盖鱼(Anoplopoma fimbria)是一种名贵的冷水经济鱼类,已在我国北方地区开展小规模的工厂化繁育和养成.目前,我国关于裸盖鱼的病害研究仍是空白.本研究对山东烟台一养殖场自然发生疥疮病的裸盖鱼进行了病原分析,从发病鱼体内分离得到形态一致的优势菌株,命名为AF-1,并对其进行了致病性检测、菌种鉴定及药物敏感性研究.人工感染实验证明,AF-1对裸盖鱼有致病性,呈现症状与自然发病状态一致;结合形态观察、革兰氏染色、生理生化特征、16S rRNA和gyrB基因序列进化树分析,将AF-1鉴定为杀鲑气单胞菌(Aeromonas salmonicida);药物敏感性实验结果显示,AF-1对青霉素、阿莫西林、头孢噻吩等13种抗生素具有耐药性,对氟苯尼考、氟甲喹等16种抗生素敏感.综上所述,本研究首次报道了我国养殖裸盖鱼感染杀鲑气单胞菌病例,为裸盖鱼养殖过程中的疾病防控和疫苗开发提供了参考.     

Effects of different feeding regimes on juvenile black rockfish (Sebastes schlegilii) survival,growth, digestive enzyme activity,body composition and feeding costs

The objective of this study was to establish a proper feeding regime for juvenile black rockfish (Sebastes schlegilii) intended for stock enhancement. We used a combined experimental design to assess the effects of different feeding rates (1%, 3% and 5% body weight per day (BW/day)) and feeding frequencies (apparent satiety or restricted feeding 1 to 3 times daily) on survival, growth, digestive enzyme activity, body composition and feeding costs over 50 days, from newly weaned (initial average weight 1.5 ± 0.2 g) to release size. The results showed that feeding rate significantly affected fish growth, feed utilization and body composition. The highest feed conversion ratio was at 3% BW/day. Increased feeding rates resulted in enhanced growth and higher body lipid content, but this was also accompanied by decreased contents of moisture, protein and ash. With feeding to apparent satiety, increased feeding frequency resulted in higher growth performance and enhanced intestinal lipase activity but decreased gastric protease activity. There were no growth improvements or effects on digestive enzyme activity and body composition with increasing feeding frequency (from one to three times daily) under a fixed‐quantify feeding (3% BW/day). Feed costs increased with increasing feeding rates or apparent satiety feeding frequencies and were associated with shorter growth times to reach a suitable size for release. Our study suggests that a feeding rate of 3% BW/day is the best feeding regime for proper growth and reduced feed costs in juvenile black rockfish before releasing for stock enhancement.     

The development of live vaccines for furunculosis lacking the A-layer and O-antigen of Aeromonas salmonicida

Abstract. Mutants of Aeromonas salmonicida strains lacking either the A-protein, O-antigen or both of these major surface antigens were tested in rainbow trout, Oncorhynchus mykiss (Walbaum), for their suitability as live vaccines (LV). All of these mutants were shown to be attenuated, as fish receiving ∼5 × 10⁷ of the respective strains showed no clinical signs of furunculosis. Immersion vaccination of fish in 5 × 10⁷ cfu ml^-1 of these strains with an identical immersion dose 14 days later resulted in significant protection by all strains from challenge with a heterologous virulent strain of A. salmonicida 5 weeks later. The levels of protection conferred were all greater than or equal to that provided by an injected bacterin using the same vaccination schedule. With one exception, all LV strains that still possessed a functional O-antigen provided protective indices (PI) four- to seven-fold greater than the PI for the fish injected with bacterin. When antibody responses of vaccinated fish were compared, it was found that only vaccination by bacterin gave rise to a measurable agglutinating litre. Western immunoblots using the immune fish sera failed to reveal any major differences in antigen recognition in fish that received any of the vaccines tested. These data suggest that the immune response generated by the use of live vaccine strains is different from that generated by a bacterin, and that these useful mutations may be incorporated into existing furunculosis LVs for further attenuation.     

Transmission of protease genes into a protease-deficient mutant of Aeromonas salmonicida in river sediments

Abstract. The transformation of Aeromonas salmonicida with DNA fragments from bacterial cell-free sonicates was investigated with intraspecific, interspecific band intergeneric fish pathogenic bacteria including Aeromonas salmonicida, Aeromonas hydrophila, Pseiidomonas fluorescens and Vibrio anguillarum strains as donor bacteria. A phenotypic marker for transformation was extracellular protease production since a protease-deficient mutant NTG-1 induced from pathogenic A. salmonicida strain A-7301 by mutagenesis was used as a recipient. This mutant was non-pathogenic to rainbow trout. The mutant was incubated with each sonicate at 20°C for 20 days with a nutrient-poor medium containing a trace (5 μg/ml each) of both humic acid and tryptone in the presence of clean river sand (100 g/100 ml medium) corresponding with an environment of rivers. During the incubation, the survival of mutant NTG-1 cells was observed and protease positive NTG-1 cells were isolated from each culture. The protease production of the isolates was due to the transmission of protease genes of the donor strains. The activity of proteases produced by the transformants extra-cellularly was determined. These transformants induced with the sonicates of the parent strain, intraspecific strain and with the sonicates of the interspecific A. hydrophila strain were pathogenic to rainbow trout, whereas the transformants derived with the sonicates of the intergeneric strains P. fluorescens and V. anguiUarum showed non-pathogenicity, although all the donor strains, with the exception of the P. fluorescens strain, were pathogenic. These findings are interesting since they demonstrate that trausformation in A. salmonicida occurs with considerable ease even intergenencally and interspecifically, as well as intraspecifically in river environments, and that there is a large difference in the lethal toxicity of extracellular protease produced by these bacteria.