克氏原螯虾源维氏气单胞菌的分离鉴定及组织病理学观察

ISOLATION, IDENTIFICATION AND PATHOHISTOLOGICAL OBSERVATION OF AEROMONAS VERONII FROM PROCAMBARUS CLARKII

  • 摘要: 为确定克氏原螯虾(Procambarus clarkii)暴发性死亡的病原体, 研究从一例患病克氏原螯虾的肝胰腺分离到了一株优势菌株QD160502, 根据菌株形态、生理生化特性, 16S rDNA及gyrB序列分析, 将其鉴定为维氏气单胞菌(Aeromonas veronii)。回归感染实验证明, 该分离株可使克氏原螯虾出现与自然发病相同症状。组织病理学观察可见克氏原螯虾肝胰腺、肠道与心脏组织病理损伤, 且随感染持续时间而逐渐加重, 在感染12h后肝小管分泌细胞中出现内含棕色颗粒的转运小泡; 感染24h后肝小管间炎性细胞浸润, 肠道结缔组织萎缩, 心脏组织出现空泡样扩张; 感染48h后肝小管储存细胞与分泌细胞大量解体并空泡化, 肠道上皮皱嵴减少, 心肌空泡样扩张增加; 感染72h后肝小管和肠道上皮细胞坏死, 且在心脏组织中发现透明血细胞聚集。药物敏感性检测发现QD160502菌株对恩诺沙星、诺氟沙星、四环素、强力霉素等敏感, 但对青霉素、链霉素、卡那霉素等抗生素耐药。研究为克氏原螯虾的健康养殖和细菌性疾病的防控提供指导依据。

     

    Abstract: To identify the aetiology of the outbreak of the disease in cultured Procambarus clarkii, we isolated a dominant strain of bacteria (QD160502) from the hepatopancreas of diseased Procambarus clarkii. It was identified as Aeromonas veronii based on morphological observation, biochemical identification, physiology characteristics and sequencing of 16S rDNA and gyrase B subunit gene. The symptoms of P. clarkii artificially infected with strain QD160502 were similar with those of natural infected P. clarkii. The pathological changes were observed by HE staining. The histological changes of P. clarkii caused by A. veronii infection were gradually aggravated. 12 hours after challenge, some transferred vacuoles containing granular materials were found in hepatopancreas blasenzenllen cells. 24 hours after challenge, inflammatory cells infiltration was observed in hepatopancreas, and intestinal connective tissue atrophy and vacuole-like dilations were observed in myocardial tissue. 48 hours after challenge, mass of hepatopancreas restzellen cells and blasenzenllen cells disintegration, intestinal mussel flods decreased and vacuole-like dilations increased in the myocardial tissue. 72 hours after challenge, hepatopancreas and intestinal epithelial cells became necrotic and damaged, and hyaline blood cells aggregation were observed in myocardial tissue. The antimicrobial sensitivity test showed that the strain QD160502 was highly sensitive to enrofloxacin, norfloxacin, tetracycline, doxycycline, etc, but resistant to penicillin, streptomycin, kanamycin, etc. This study provides theoretical basis for the prevention and treatment of A. veronii infection diseases in P. clarkii.

     

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