1株鲫鱼致病性腐败希瓦氏菌的分离鉴定及中草药抑菌效果评价OA
Isolation,Identification of a Pathogenic Shewanella putrefaciens from Crucian Carp,and Evaluation of Antibacterial Effect of Chinese Herbal Medicine
[目的]探究江苏金坛某养殖场鲫鱼病害的发病原因,从而为后续鲫鱼养殖及相关疾病的防治提供参考.[方法]采集发病鲫鱼内脏样品进行细菌分离纯化,通过形态学观察、电镜观察和生理生化测定对其进行初步鉴定,进一步结合16S rDNA基因及系统发育树分析确定其种属关系,并经回归感染试验检测其致病性.采用纸片扩散法和琼脂打孔法分别进行药敏试验和中草药抑菌试验,筛选有效治疗药物.[结果]分离菌株在LB培养基中长出边缘光滑菌落,呈现乳白色隆起,无溶血性;革兰染色为阴性,透射电镜下呈杆状.16S rDNA序列分析显示,分离菌株与腐败希瓦氏菌(Shewanella putrefaciens)菌株Hammer 95(NR_044863.1)相似性达99.17%,在系统发育树上与腐败希瓦氏菌ATCC 8071(NR_119141.1)自然聚为一支,确定该分离菌株为腐败希瓦氏菌.致病性试验结果显示,从感染鲫鱼中重新分离到与接种菌株相同的细菌.药敏试验及中草药抑菌试验结果显示,分离株对氨苄西林、米诺环素和阿米卡星等28种抗菌药及五倍子(Galla chinensis)、乌梅(Prunus mume)等8味中草药表现敏感,其中五倍子(Galla chinensis)和诃子(Chebula retz)抑菌效果最佳,最小抑菌浓度(MIC)与最小杀菌浓度(MBC)均为15.63 mg/mL,且五倍子、诃子、丁香(Syzygium aromaticum)具有协同增效作用.[结论]本研究证实了腐败希瓦氏菌是导致江苏金坛某养殖场鲫鱼感染死亡的主要病原菌,且该菌对氨苄西林、米诺环素等28种抗菌药物及五倍子、诃子等8味中草药敏感.本试验结果为腐败希瓦氏菌感染的科学防治提供了理论依据.
[Objective]This experiment was to explore the causes of crucian carp diseases in a fish farm in Jintan,Jiangsu province,so as to provide references for subsequent crucian carp breeding and the prevention and control of related diseases.[Method]Internal organ samples of diseased crucian carp were collected for bacterial isolation and purification.The isolated bacteria were preliminarily identified through morphological observation,electron microscopy observation and physiological and biochemical determination.The species relationship was determined by combining the 16S rDNA gene sequence and phylogenetic tree analysis.The pathogenicity of the isolated bacteria was detected by the regression infection test.Drug sensitivity test and antibacterial tests of Chinese herbal medicines were conducted by the disk diffusion method and the agar perforation method,respectively,to screen effective therapeutic drugs.[Result]The colonies of the isolated strain had smooth edges in the LB medium,presenting as milky white protrusions and were not hemolytic.Gram staining was negative and it appeared rod-shaped under transmission electron microscope.16S rDNA sequence analysis revealed that the isolated strain had a similarity of 99.17%to the Shewanella putrefaciens strain Hammer 95(NR_044866.1).On the phylogenetic tree,it naturally aggregated into the same branch with Shewanella putrefaciens ATCC 8071(NR_119141.1).It was determined that the isolated strain was Shewanella putrefaciens.The results of pathogenicity test showed that the same bacteria as the inoculated strain were re-isolated from the infected crucian carp.The results of drug sensitivity test and antibacterial test of Chinese herbal medicines showed that the isolated strain was sensitive to 28 kinds of antibacterial drugs such as ampicillin,minocycline,and amicacin,as well as 8 kinds of Chinese herbal medicines such as Galla chinensis and Prunus mume.Among them,Galla chinensis and Chebula retz had the best antibacterial effects,with both minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of 15.63 mg/mL.Moreover,Galla chinensis,Chebula retz and Syzygium aromaticum had a synergistic effect.[Conclusion]This study confirmed that Shewanella putrefaciens was the primary pathogen responsible for the infection and mortality of farmed crucian carp in Jintan,Jiangsu.Moreover,the isolated strain was sensitive to 28 kinds of antibacterial drugs such as ampicillin and minocycline,as well as 8 kinds of Chinese herbal medicines such as Galla chinensis and Chebula retz.This results provided a theoretical basis for the scientific prevention and control of Shewanella putrefaciens infection
孙立;段蕊;姜虎成;薛晖;赵彦华
江苏省淡水水产研究所,南京 210017||江苏海洋大学,连云港 222005江苏海洋大学,连云港 222005江苏省淡水水产研究所,南京 210017江苏省淡水水产研究所,南京 210017江苏省淡水水产研究所,南京 210017
农业科技
鲫鱼腐败希瓦氏菌药敏试验致病性中草药
crucian carpShewanella putrefaciensdrug sensitivity testpathogenicityChinese herbal medicine
《中国畜牧兽医》 2026 (3)
1573-1584,12
国家大宗淡水鱼产业技术体系(CARS-45)
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