1. Tianjin Fishery Research Institute, Tianjin 300221, China; 2. Tianjin Sanyuan Baodi Agricultural Technology Co., Ltd, Tianjin 301823, China; 3. Agricultural Development Service Center of Beichen District, Tianjin 300400, China
Abstract:In order to explore the pathogenicity and environmental impact factors of yeast Metschnikowia bicuspidate(JMB-1), and to screen pathogenic sensitive drugs for control of “milky disease” in Chinese mitten crab Eriocheir sinensis, the median lethal concentration (LC50) of JMB-1 isolated from the diseased crab was determined in healthy Chinese mitten crab with body weight of (6.8±0.7) g by challenge experiments, the effect of temperature on infection was studied, and the pathogenicity of JMB-1 was explored. The sensitivity of JMB-1 to drugs was tested by Oxford cup plate assay, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of preferred drugs were tested. The results showed that the median lethal concentration (LC50) of JMB-1 was 1.05×105 cfu/mL in 14 days of infection at water temperature of 15 ℃, with positive correlation between the mortality of Chinese mitten crab and temperature in the range of 10—30 ℃. Challenge test revealed that JMB-1 concentration of 2.1×103 cfu/mL led to 90% mortality of Chinese mitten crabin 21 days under laboratory culture conditions (22 ℃). Chlorhexidine gluconate solution, nystatin and benzalkonium had a better inhibitory effect on JMB-1, with an MIC of 10, 10, 5 mg/L and an MBC of 20, 100, 10 mg/L, respectively. The crude extract of garlic also had a good inhibitory effect on JMB-1, and a dilution of 32 times killed JMB-1. The finding provides reference with the scientific prevention and control of "milk disease" in Chinese mitten crab.
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