Killing effect of compound peroxymonosulphate powder on four common aquatic pathogens
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摘要: 为验证过硫酸氢钾复合物粉在水产养殖中消毒的有效性，评价了其对草鱼 (Ctenopharyngodon idella) 呼肠孤病毒 (Grass carp reovirus, GCRV)、斑点叉尾鮰 (Ictalurus punctatus) 病毒 (Channel catfish virus, CCV)、嗜水气单胞菌 (Aeromonas hydrophila) 和无乳链球菌 (Streptococcus agalactiae) 4种水产常见致病病原的杀灭效果，以及有机物、温度、pH 3种因素对杀菌效果的影响，并在此基础上开展了过硫酸氢钾复合物粉对池塘养殖水体中目标菌的杀灭试验，以及对患嗜水气单胞菌的病鲫 (Carassius auratus) 的药效试验。结果显示，过硫酸氢钾复合物粉对4种致病微生物均具有显著的杀灭效果。降低菌悬液中有机物浓度、适当提高作用温度或适当降低消毒液pH，均能促进过硫酸氢钾复合物粉消毒液的杀菌效果。通过目标菌的杀灭试验和药效试验，进一步确定了该消毒剂在实际使用中对目标菌杀灭的有效性和对鲫感染嗜水气单胞菌后的显著治疗效果。研究表明，过硫酸氢钾复合物粉是潜在的水产用消毒剂。Abstract: To study the disinfection validity of compound peroxymonosulfate powder in aquaculture, we evaluated its killing effect on grass carp (Ctenopharyngodon idella) reovirus (GCRV), channel catfish (Ictalurus punctatus) virus (CCV), Aeromonas hydrophila and Streptococcus agalactiae, and studied the germicidal effects of three factors (Organic matter, temperature and pH). Moreover, we conducted a killing assay of the target bacteria in pond aquaculture water and a pharmacodynamic test of crucian carp (Carassius auratus) with A. hydrophila disease. The results show that the compound peroxymonosulfate powder had a significant killing effect on the four pathogens. Reducing organic matter concentration in bacterial suspension, appropriately increasing temperature or decreasing pH of disinfectant could promote the germicidal effect. In addition, through the killing assay of target bacteria and pharmacodynamic test of crucian carp, we further confirmed the disinfectant validity on target bacteria and significant therapeutic effect of crucian carp infected with A. hydrophila. Thus, the study indicates that compound peroxymonosulfate powder can be used as a potential candidate for disinfectant in aquaculture.
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Key words:
- Compound peroxymonosulfate powder /
- Disinfection /
- Virus inactivation /
- Germicidal effect
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图 1 中和剂、中和产物和消毒剂对细胞生长的影响
注：a. 草鱼肾细胞，正常组；b. CIK，中和剂组；c. 草鱼肾细胞，中和产物组；d. 草鱼肾细胞，消毒剂组；e. 斑点叉尾鮰卵巢细胞，正常组；f. 斑点叉尾鮰卵巢细胞，中和组；g. 斑点叉尾鮰卵巢细胞，中和产物组；h. 斑点叉尾鮰卵巢细胞，消毒剂组。
Figure 1. Effects of neutralizer, neutralization products and disinfectant on cell growth
Note: a. CIK, normal group; b. CIK, neutralizer group; c. CIK, neutralization product group; d. CIK, disinfectant group; e. CCO, normal group; f. CCO, neutralizer group; g. CCO, neutralization product group; h. CCO, disinfectant group.
图 2 过硫酸氢钾复合物粉对呼肠孤病毒和斑点叉尾鮰病毒的灭活结果
注：a. 呼肠孤病毒；b. 斑点叉尾鮰病毒；与消毒前原始病毒滴度相比，*. P <0.01。
Figure 2. Inactivation results of GCRV and CCV by compound peroxymonosulphate powder
Note: a. GCRV; b. CCV; when compared to the original virus titer before disinfection, *. P <0.01.
图 4 有机物浓度对过硫酸氢钾复合物粉杀菌效果的影响
注：0%、10%、25%和50%分别代表有机物在菌悬液中的体积分数；a. 嗜水气单胞菌；b. 无乳链球菌；与消毒前原始细菌数相比，*. P <0.01；#. 0.01<P<0.05；图5同此。
Figure 4. Germicidal effect of organic matter concentration on compound peroxymonosulphate powder
Note: 0%, 10%, 25% and 50% represent the volume fractions of organic matter in the bacterial suspension, respectively; a. A. hydrophila; b. S. agalactiae; when compared to the original number of bacteria before disinfection, *. P <0.01; #. 0.01<P<0.05. The same case in Fig. 5.
图 5 温度对过硫酸氢钾复合物粉杀菌效果的影响
Figure 5. Germicidal effect of temperature on compound peroxymonosulphate powder
图 6 pH对过硫酸氢钾复合物粉杀菌效果的影响
Figure 6. Germicidal effect of pH on compound peroxymonosulphate powder
图 7 养殖水中目标菌的杀灭效果
注：a. 阴性对照组；b. 嗜水气单胞菌；c. 无乳链球菌；与消毒前原始细菌数相比，*. P <0.01。
Figure 7. Killing effect of target bacteria in aquaculture water
Note:a. Negative control group; b. A. hydrophila; c. S. agalactiae; when compared to the original number of bacteria before disinfection, *. P <0.01.
图 8 过硫酸氢钾复合物粉提高鲫感染嗜水气单胞菌后的成活率
注：与阳性对照组成活率相比，*. P<0.01；#. 0.01<P<0.05。
Figure 8. Increased survival rate of C. auratus infected with A. hydrophila after treatment of compound peroxymonosulphate powder
Note: When compared to the survival rate of the positive control group, *. P <0.01; #. 0.01<P<0.05.
表 1 正式试验
Table 1. Formal test
组别
Group类目
Category1 (0.45 mL 消毒剂+0.05 mL 病毒悬液) +4.5 mL 无菌水 2 (0.45 mL 消毒剂+0.05 mL 病毒悬液) +4.5 mL 中和剂 3 (0.45 mL 无菌水+0.05 mL 病毒悬液) +4.5 mL 中和剂 4 (0.45 mL 消毒剂+4.5 mL 中和剂) + 0.05 mL 病毒悬液 5 4.95 mL 无菌水+0.05 mL 病毒悬液 6 细胞正常培养 
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表 2 中和剂鉴定的组别
Table 2. Groups of neutralizer identification
组别
Group类目
Category1 (0.45 mL 消毒剂+0.05 mL 菌悬液) +4.5 mL PBS 2 (0.45 mL 消毒剂+0. 05 mL 菌悬液) +4.5 mL 中和剂 3 (0.45 mL PBS+0.05 mL 菌悬液) +4.5 mL 中和剂 4 (0. 45 mL 消毒剂+4.5 mL 中和剂) +0.05 mL 菌悬液 5 4.95 mL PBS+0.05 mL 菌悬液 6 中和剂+PBS
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表 3 病毒灭活试验的中和剂鉴定结果
Table 3. Neutralizer identification results of virus inactivation assay
组别
GroupTCID50对数值 lg 呼肠孤病毒
GCRV斑点叉尾鮰病毒
CCV1 1.11±0.16 0 2 1.44±0.08 0 3 4.41±0.13 4.50±0.22 4 4.50±0.22 4.39±0.08 5 4.41±0.13 4.59±0.13 6 0 0 注：该表组别与表1对应。 Note: The groups in this table correspond to the groups in Table 1.
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表 4 用于杀菌试验的中和剂鉴定结果
Table 4. Neutralizer identification results of bactericidal assay
组别
Group嗜水气单胞菌质量浓度
Mass concentration of
A. hydrophila/
(CFU∙mL−1)无乳链球菌质量浓度
Mass concentration of
S. agalactiae/
(CFU∙mL−1)1 0 0 2 (3.5±0.82)×105 58±18 3 (3.15±0.72)×106 (2.27±0.31)×106 4 (2.68±0.26)×106 (2.33±0.38)×106 5 (2.85±0.33)×106 (2.03±0.20)×106 6 0 0 注：该表组别与表2对应。 Note: The groups in this table correspond to the groups in Table 2.
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表 5 过硫酸氢钾复合物粉定性杀菌试验
Table 5. Qualitative suspension test of compound peroxymonosulphate powder
菌种Bacteria 最低杀菌有效稀释度 Minimal bactericidal effective concentration 5 min 10 min 15 min 30 min 60 min 嗜水气单胞菌 A. hydrophila 1∶400 1∶400 1∶800 1∶800 1∶3 200 无乳链球菌 S. agalactiae 1∶400 1∶400 1∶800 1∶1 600 1∶3 200
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