Isolation, identification and pathogenicity analysis of pathogenic bacterium P5W in Sebastes schlegelii
CHEN Yuzhen, WANG Tengteng, HAN Huizong, CUI Guangxin, WANG Fei, ZHANG Mingliang, JIANG Haibin
Sebastes schlegelii is the main fish for proliferation and release and deep-water cage aquaculture in the coastal areas of northern China. In recent years, with the expansion of the scale of S. schlegelii farming and the limitations of disease prevention and control techniques, aquaculture diseases occur frequently. To find out the cause of the disease and death of S. schlegelii during culture, a dominant strain P5W was isolated from the focus of diseased S. schlegelii in Yantai aquaculture area. Through morphological characteristics observation, mitochondrial 16S rDNA sequence comparison and phylogenetic evolutionary identification, the strain P5W was identified as Vibrio harveyi. The artificial infection experiment, histopathological observation, extracellular product enzymatic and hemolytic activity analysis, physiological biochemical and susceptibility testing were further carried out. Healthy S. schlegelii was then challenged with the strain P5W through intraperitoneal injection. The results confirmed that P5W was the pathogenic bacterium of S. schlegelii, causing ulceration and congestion on the surface of challenged S. schlegelii. The median lethal dose of P5W was 5.7×106 CFU/mL. Histopathological analysis revealed that the liver, spleen, kidney and muscle showed different degrees of lesions and necrosis. For example, hepatocytes were obviously vacuolized, blood cells diffused in spleen, kidney tubule partial epithelial were shed and necrotic and muscle fiber fractured. The extracellular products of the bacterium had the activities of protease, lipase, lecithin, amylase, urease and gelatinase, and the hemolytic activity of sheep erythrocytes. P5W could survive at temperatures ranging from 15 to 35℃, acidities varying between pH 5 and 10, NaCl content (w/v%) ranging from 1 to 8. The best condition for P5W was 25℃, pH 7, and NaCl content varying between 2 and 4. The susceptibility testing showed that P5W was particularly sensitive to 13 drugs including cefotaxime, ceftriaxone, levofloxacin, tetracycline and cotrimoxazole. The research results provided a theoretical and practical guidance for the prevention and control of vibriosis during the culture of S. schlegelii. As a strong pathogenic strain of V. harveyi, P5W can be used to selecting and breeding anti-disease seedlings and developing a V. harveyi vaccine for S. schlegelii.