Laboratory Animal and Comparative Medicine ›› 2023, Vol. 43 ›› Issue (4): 347-354.DOI: 10.12300/j.issn.1674-5817.2023.060
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Ying HUANG()(
), Siyu WEI, Li CAI, Sujing QIANG, Dongting LI, Yuqiang DING
Received:
2023-05-09
Revised:
2023-08-03
Online:
2023-08-25
Published:
2023-08-25
Contact:
Ying HUANG
CLC Number:
Ying HUANG,Siyu WEI,Li CAI,et al. Microbiological Monitoring Analysis of Laboratory Rats and Mice from Vendors: Department of Laboratory Animal Science of Fudan University as an Example[J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 347-354. DOI: 10.12300/j.issn.1674-5817.2023.060.
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细菌 Bacteria | 采样 动物 Animal | 革兰染色 Gram staining | 培养 物质 Sample | 第一天接种 Inoculation on day 1 | 第二天再接种 Inoculation on day 2 | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|
培养基 Culture medium | 37 ℃ 培养24 h后的典型菌落特点 Colony characteristics after 24 h at 37 ℃ | 培养基 Culture medium | 37 ℃培养24 h后是否β-溶血 β-Hemolytic reaction after 24 h at 37 ℃ | ||||||||
直径 Diameter/mm | 颜色/色泽 Color/luster | 溶血性 Hemolysis | 其他 Other | ||||||||
支气管鲍特杆菌 Bordetella bronchiseptica | 大鼠 | - | 呼吸道分泌物 | 血平板 | 1 | 灰白色 | 轻微α-溶血 | / | 血平板 | 否 | |
DHL平板 | 2 | 无色半透明 | / | / | |||||||
鼠棒状杆菌 Corynebacterium kutscheri | 大鼠、小鼠 | + | 呼吸道分泌物 | 血平板 | 1 | 白色无光泽 | γ-溶血 | 触之较硬,不易乳化 | 血平板 | 否 | |
嗜肺巴斯德杆菌 Pasteurella pneumotropica | 大鼠、小鼠 | - | 呼吸道分泌物 | 血平板 | 0.5~1 | 灰白色或露滴样 | γ-溶血,或轻微α-溶血 | 聚集时质地似奶油 | 血平板 | 否 | |
沙门菌 Salmonella spp. | 大鼠、小鼠 | - | 回盲肠部内容物 | DHL平板 | 2 | 无色半透明,带黑心或全黑 | / | 表面光滑湿润 | 血平板 | 否 | |
金黄色葡萄球菌 Staphylococcus aureus | 大鼠、小鼠 | + | 回盲肠部内容物 | 高盐 甘露醇 | 1 | 黄色凸起 | / | 周边培养基从红色变黄色 | 血平板 | 是(白或金黄圆形凸起菌落) | |
肺炎克雷伯菌 Klebsiella pneumoniae | 大鼠、小鼠 | - | 回盲肠部内容物 | DHL平板 | 1~5 | 淡粉色,大而隆起 | / | 光滑湿润,呈黏液脓汁状 | 血平板 | 否 | |
绿脓杆菌 Pseudomonas aeruginosa | 大鼠、小鼠 | - | 回盲肠部内容物 | NAC液 | 1~3 | 浑浊均匀生长 | / | 菌落扁平,可产绿色色素 | NAC平板 | / |
Table 1 Colony characteristics and Gram nature of bacteria excluded in SPF mice/rats on nutrient agar plates
细菌 Bacteria | 采样 动物 Animal | 革兰染色 Gram staining | 培养 物质 Sample | 第一天接种 Inoculation on day 1 | 第二天再接种 Inoculation on day 2 | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|
培养基 Culture medium | 37 ℃ 培养24 h后的典型菌落特点 Colony characteristics after 24 h at 37 ℃ | 培养基 Culture medium | 37 ℃培养24 h后是否β-溶血 β-Hemolytic reaction after 24 h at 37 ℃ | ||||||||
直径 Diameter/mm | 颜色/色泽 Color/luster | 溶血性 Hemolysis | 其他 Other | ||||||||
支气管鲍特杆菌 Bordetella bronchiseptica | 大鼠 | - | 呼吸道分泌物 | 血平板 | 1 | 灰白色 | 轻微α-溶血 | / | 血平板 | 否 | |
DHL平板 | 2 | 无色半透明 | / | / | |||||||
鼠棒状杆菌 Corynebacterium kutscheri | 大鼠、小鼠 | + | 呼吸道分泌物 | 血平板 | 1 | 白色无光泽 | γ-溶血 | 触之较硬,不易乳化 | 血平板 | 否 | |
嗜肺巴斯德杆菌 Pasteurella pneumotropica | 大鼠、小鼠 | - | 呼吸道分泌物 | 血平板 | 0.5~1 | 灰白色或露滴样 | γ-溶血,或轻微α-溶血 | 聚集时质地似奶油 | 血平板 | 否 | |
沙门菌 Salmonella spp. | 大鼠、小鼠 | - | 回盲肠部内容物 | DHL平板 | 2 | 无色半透明,带黑心或全黑 | / | 表面光滑湿润 | 血平板 | 否 | |
金黄色葡萄球菌 Staphylococcus aureus | 大鼠、小鼠 | + | 回盲肠部内容物 | 高盐 甘露醇 | 1 | 黄色凸起 | / | 周边培养基从红色变黄色 | 血平板 | 是(白或金黄圆形凸起菌落) | |
肺炎克雷伯菌 Klebsiella pneumoniae | 大鼠、小鼠 | - | 回盲肠部内容物 | DHL平板 | 1~5 | 淡粉色,大而隆起 | / | 光滑湿润,呈黏液脓汁状 | 血平板 | 否 | |
绿脓杆菌 Pseudomonas aeruginosa | 大鼠、小鼠 | - | 回盲肠部内容物 | NAC液 | 1~3 | 浑浊均匀生长 | / | 菌落扁平,可产绿色色素 | NAC平板 | / |
微生物检测指标 Pathogens tested | 检测数 No. of tested animals | 合格数 (合格率/%) No. of qualified animals (rate/%) | 检出数 (检出率/%) No. of positive animals (rate/%) | |
---|---|---|---|---|
寄生虫指标(镜检)Parasites (microscopic examination) | ||||
体外In vitro | 节肢动物 Ectoparasites | 158 | 158 (100) | 0 (0) |
体内In vivo | 蠕虫 All Helminths | 158 | 158 (100) | 0 (0) |
鞭毛虫 Flagellates | 158 | 158 (100) | 0 (0) | |
纤毛虫 Ciliates | 158 | 158 (100) | 0 (0) | |
细菌学指标(培养/分子生物学方法)Bacteria ( culture/molecular biology methods) | ||||
支气管鲍特杆菌 Bordetella bronchiseptica * | 19 | 19 (100) | 0 (0) | |
鼠棒状杆菌 Corynebacterium kutscheri | 158 | 158 (100) | 0 (0) | |
嗜肺啮齿杆菌 Rodentibacter pneumotropicus | 158 | 158 (100) | 0 (0) | |
啮齿杆菌H型 Rodentibacter heylii | 158 | 157 (99.37) | 1 (0.63) | |
沙门菌 Salmonella spp. | 158 | 158 (100) | 0 (0) | |
金黄色葡萄球菌 Staphylococcus aureus | 158 | 141 (89.24) | 17 (10.76) | |
肺炎克雷伯杆菌 Klebsiella pneumoniae | 158 | 154 (97.47) | 4 (2.53) | |
绿脓杆菌 Pseudomonas aeruginosa | 158 | 153 (96.84) | 5 (3.16) | |
病毒等指标(酶联免疫吸附实验)Viruses, etc (ELISA) | ||||
汉坦病毒 Hantavirus * | 19 | 19 (100) | 0 (0) | |
小鼠肝炎病毒 Mouse hepatitis virus # | 139 | 139 (100) | 0 (0) | |
鼠痘病毒 Ectromelia virus # | 139 | 139 (100) | 0 (0) | |
仙台病毒 Sendai virus | 158 | 154 (97.47) | 4 (2.53) | |
小鼠肺炎病毒 Pneumonia virus of mice | 158 | 158 (100) | 0 (0) | |
呼肠孤病毒III型 Reovirus type 3 | 158 | 158 (100) | 0 (0) | |
小鼠细小病毒 Minute virus of mice # | 139 | 139 (100) | 0 (0) | |
大鼠细小病毒Rat parvovirus * | RV株 | 19 | 19 (100) | 0 (0) |
H-1株 | 19 | 19 (100) | 0 (0) | |
大鼠冠状病毒 Rat coronavirus * | 19 | 19 (100) | 0 (0) | |
支原体 Mycoplasma spp. | 158 | 158 (100) | 0 (0) | |
泰泽病原体 Tyzzer's organism | 158 | 158 (100) | 0 (0) | |
弓形体 Toxoplasma gondii | 158 | 158 (100) | 0 (0) |
Table 2 Pathogen test results of SPF mice/rats from vendors during 2021-2023 according to national standards
微生物检测指标 Pathogens tested | 检测数 No. of tested animals | 合格数 (合格率/%) No. of qualified animals (rate/%) | 检出数 (检出率/%) No. of positive animals (rate/%) | |
---|---|---|---|---|
寄生虫指标(镜检)Parasites (microscopic examination) | ||||
体外In vitro | 节肢动物 Ectoparasites | 158 | 158 (100) | 0 (0) |
体内In vivo | 蠕虫 All Helminths | 158 | 158 (100) | 0 (0) |
鞭毛虫 Flagellates | 158 | 158 (100) | 0 (0) | |
纤毛虫 Ciliates | 158 | 158 (100) | 0 (0) | |
细菌学指标(培养/分子生物学方法)Bacteria ( culture/molecular biology methods) | ||||
支气管鲍特杆菌 Bordetella bronchiseptica * | 19 | 19 (100) | 0 (0) | |
鼠棒状杆菌 Corynebacterium kutscheri | 158 | 158 (100) | 0 (0) | |
嗜肺啮齿杆菌 Rodentibacter pneumotropicus | 158 | 158 (100) | 0 (0) | |
啮齿杆菌H型 Rodentibacter heylii | 158 | 157 (99.37) | 1 (0.63) | |
沙门菌 Salmonella spp. | 158 | 158 (100) | 0 (0) | |
金黄色葡萄球菌 Staphylococcus aureus | 158 | 141 (89.24) | 17 (10.76) | |
肺炎克雷伯杆菌 Klebsiella pneumoniae | 158 | 154 (97.47) | 4 (2.53) | |
绿脓杆菌 Pseudomonas aeruginosa | 158 | 153 (96.84) | 5 (3.16) | |
病毒等指标(酶联免疫吸附实验)Viruses, etc (ELISA) | ||||
汉坦病毒 Hantavirus * | 19 | 19 (100) | 0 (0) | |
小鼠肝炎病毒 Mouse hepatitis virus # | 139 | 139 (100) | 0 (0) | |
鼠痘病毒 Ectromelia virus # | 139 | 139 (100) | 0 (0) | |
仙台病毒 Sendai virus | 158 | 154 (97.47) | 4 (2.53) | |
小鼠肺炎病毒 Pneumonia virus of mice | 158 | 158 (100) | 0 (0) | |
呼肠孤病毒III型 Reovirus type 3 | 158 | 158 (100) | 0 (0) | |
小鼠细小病毒 Minute virus of mice # | 139 | 139 (100) | 0 (0) | |
大鼠细小病毒Rat parvovirus * | RV株 | 19 | 19 (100) | 0 (0) |
H-1株 | 19 | 19 (100) | 0 (0) | |
大鼠冠状病毒 Rat coronavirus * | 19 | 19 (100) | 0 (0) | |
支原体 Mycoplasma spp. | 158 | 158 (100) | 0 (0) | |
泰泽病原体 Tyzzer's organism | 158 | 158 (100) | 0 (0) | |
弓形体 Toxoplasma gondii | 158 | 158 (100) | 0 (0) |
Figure 1 Representative images of positive bacteria or parasites detected in SPF mice/rats from vendors during 2021-2023Note:a-c, Colonies of Enterobacter spp., Klebsiella oxytoca, and Klebsiella pneumoniae growth on DHL agar plates,respectively; d-e, Colonies of Staphylococcus aureus and Rodentibacter heylii growth on blood plates, respectively; f, Colonies of Pseudomonas aeruginosa growth on NAC agar plates; g, Image of Entamoeba muris in the ileocecal region. Triangular arrow indicates the bacterial colony or amoeba body. Scale bar=2 mm (a-f), 10 μm (g).
项目 Pathogens | SD 大鼠 SD rats | 近交系小鼠 Inbred mice | 远交系小鼠 Outbred mice | 免疫缺陷小鼠 Immunodeficient mice | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
检测数No. of tested animals | 检出数(率/%) No. of positive (rate/%) | 合格率/% Rate of qualified/% | 检测数No. of tested animals | 检出数(率/%) No. of positive (rate/%) | 合格率/% Rate of qualified/% | 检测数 No. of tested animals | 检出数(率/%) No. of positive (rate/%) | 合格率/% Rate of qualified /% | 检测数 No. of tested animals | 检出数(率/%) No. of positive (rate/%) | 合格率/% Rate of qualified/% | ||||
检出总情况 Summary | 19 | 9 (47.36) | 52.63 | 87 | 15 (17.24) | 82.76 | 30 | 4 (13.33) | 86.67 | 22 | 3 (13.64) | 86.36 | |||
国家标准规定必须排除病原体 Pathogens required free of animals in national standards | |||||||||||||||
肺炎克雷伯杆菌 Klebsiella pneumoniae | 19 | 0 (0) | 100 | 87 | 2 (2.30) | 97.70 | 30 | 0 (0) | 100 | 22 | 2 (9.09) | 90.91 | |||
金黄色葡萄球菌 Staphylococcus ureus | 19 | 4 (21.05) | 78.95 | 87 | 13 (14.94) | 85.06 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
绿脓杆菌 Pseudomonas aeruginosa | 19 | 0 (0) | 100 | 87 | 0 (0) | 100 | 30 | 4 (13.33) | 86.67 | 22 | 1 (4.55) | 95.45 | |||
啮齿杆菌H型 Rodentibacter heylii | 19 | 1 (5.26) | 94.74 | 87 | 0 (0) | 100 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
仙台病毒 Sendai Virus | 19 | 4 (21.05) | 78.95 | 87 | 0 (0) | 100 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
非国标规定检测病原体 Pathogens unrequired free of animals in national standards | |||||||||||||||
阿米巴原虫 Entamoeba muris | 19 | 0 (0) | 100 | 87 | 1 (1.15) | 98.85 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
产酸克雷伯杆菌 Klebsiella oxytoca | 19 | 0 (0) | 100 | 87 | 0 (0) | 100 | 30 | 0 (0) | 100 | 22 | 1 (4.55) | 95.45 | |||
肠杆菌属 | 19 | 0 (0) | 100 | 87 | 2 (2.30) | 97.70 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
Enterobacter spp. |
Table 3 Distribution of pathogens detected in different groups of SPF mice/rats from vendors during 2021-2023
项目 Pathogens | SD 大鼠 SD rats | 近交系小鼠 Inbred mice | 远交系小鼠 Outbred mice | 免疫缺陷小鼠 Immunodeficient mice | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
检测数No. of tested animals | 检出数(率/%) No. of positive (rate/%) | 合格率/% Rate of qualified/% | 检测数No. of tested animals | 检出数(率/%) No. of positive (rate/%) | 合格率/% Rate of qualified/% | 检测数 No. of tested animals | 检出数(率/%) No. of positive (rate/%) | 合格率/% Rate of qualified /% | 检测数 No. of tested animals | 检出数(率/%) No. of positive (rate/%) | 合格率/% Rate of qualified/% | ||||
检出总情况 Summary | 19 | 9 (47.36) | 52.63 | 87 | 15 (17.24) | 82.76 | 30 | 4 (13.33) | 86.67 | 22 | 3 (13.64) | 86.36 | |||
国家标准规定必须排除病原体 Pathogens required free of animals in national standards | |||||||||||||||
肺炎克雷伯杆菌 Klebsiella pneumoniae | 19 | 0 (0) | 100 | 87 | 2 (2.30) | 97.70 | 30 | 0 (0) | 100 | 22 | 2 (9.09) | 90.91 | |||
金黄色葡萄球菌 Staphylococcus ureus | 19 | 4 (21.05) | 78.95 | 87 | 13 (14.94) | 85.06 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
绿脓杆菌 Pseudomonas aeruginosa | 19 | 0 (0) | 100 | 87 | 0 (0) | 100 | 30 | 4 (13.33) | 86.67 | 22 | 1 (4.55) | 95.45 | |||
啮齿杆菌H型 Rodentibacter heylii | 19 | 1 (5.26) | 94.74 | 87 | 0 (0) | 100 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
仙台病毒 Sendai Virus | 19 | 4 (21.05) | 78.95 | 87 | 0 (0) | 100 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
非国标规定检测病原体 Pathogens unrequired free of animals in national standards | |||||||||||||||
阿米巴原虫 Entamoeba muris | 19 | 0 (0) | 100 | 87 | 1 (1.15) | 98.85 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
产酸克雷伯杆菌 Klebsiella oxytoca | 19 | 0 (0) | 100 | 87 | 0 (0) | 100 | 30 | 0 (0) | 100 | 22 | 1 (4.55) | 95.45 | |||
肠杆菌属 | 19 | 0 (0) | 100 | 87 | 2 (2.30) | 97.70 | 30 | 0 (0) | 100 | 22 | 0 (0) | 100 | |||
Enterobacter spp. |
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