供应商来源的实验大鼠和小鼠微生物监测结果分析：以复旦大学实验动物科学部为例

doi:10.12300/j.issn.1674-5817.2023.060

摘要/Abstract

摘要：

目的对供应商来源的实验大鼠、小鼠进行常规微生物监测，为实验动物设施科学管理提供重要依据，确保相关实验结果的可靠性。方法以复旦大学实验动物科学部为例，在2021年4月到2023年4月间，按照单纯随机抽样原则，对来自7家供应商的大鼠、小鼠进行微生物质量抽检。具体参照国家标准中SPF级实验动物必须排除的微生物指标及其检测方法进行。结果抽检动物的检测总合格率为80.36%。其中SPF级大鼠抽检合格率为52.63%，SPF级近交系小鼠抽检合格率为82.76%，SPF级远交系小鼠抽检合格率为86.67%，SPF级免疫缺陷小鼠抽检合格率为86.36%。检测不合格的细菌学指标集中在金黄色葡萄球菌、绿脓杆菌、肺炎克雷伯杆菌和啮齿杆菌H型，检出率分别为10.76%、3.16%、2.53%和0.63%。不合格的血清学指标为仙台病毒，发生率为2.53%。此外，除国家标准规定SPF级小鼠必须排除的微生物指标外，还在近交系小鼠中检测到阿米巴原虫和肠杆菌属菌株；在免疫缺陷小鼠中检测到产酸性克雷伯杆菌，检出率分别为1.15%、2.30%和4.55%。结论供应商来源的大小鼠动物中存在一定的病原体感染发生率，相关动物设施有必要强化实验动物微生物监测，确保接收和饲养动物的质量，这对提高实验结果准确性和保护实验动物从业人员的职业健康具有重要意义。

关键词: 微生物, 监测, 供应商, 实验动物质量, 大鼠, 小鼠

Abstract:

Objective Conduct routine microbiological monitoring of laboratory rats and mice from vendors to provide an important basis for the scientific management of laboratory animal facility and ensure the reliability of relevant experimental data obtained from laboratory animals. Methods Taking the Department of Laboratory Animal Science of Fudan University as an example, between April 2021 and April 2023, rats and mice purchased from 7 vendors were sampled for microbiological quality according to the principle of simple random sampling on the arrival days of animal delivery. Then, surveillance tests were conducted to examine the microbiological contaminations according to the national standards of SPF laboratory animals. Results The total qualified rate was 80.36%, with 52.63% in SD rat, 82.76% in inbred mice, 86.67% in outbred mice and 86.36% in immunodeficient mice in details. The most frequent bacteria isolated were Staphylococcus aureus, Pseudomonas aeruginosa, Klebsilla pneumoniae and Rodentibacter heylii, and their detection rates were 10.76%, 3.16%, 2.53% and 0.63%, respectively. Serological assays demonstrated the highest prevalence for virus was Sendai virus, and the detection rate was 2.53%. In addition to the pathogens those must be excluded from SPF rodents, Entamoeba muris and Enterobacter spp. were also detected in inbred mice, and Klebsiella oxytoca was detected in immunodeficient mice, with the detection rates of 1.15%, 2.30% and 4.55%, respectively. Conclusion There are certain incidences of pathogen infections in laboratory rats and mice from vendors, and an efficient microbiological monitoring of laboratory animals should be implemented in animal facilities, in order to eliminate pathogen infections in laboratory animals, which is required for improving the accuracy of research results and protecting the occupational health of laboratory animal practitioners as well.

Key words: Microbiology, Monitoring, Vendor, Laboratory animal quality, Rat, Mouse

中图分类号:

Q95-33

黄缨, 韦思羽, 蔡莉, 强苏静, 李冬婷, 丁玉强. 供应商来源的实验大鼠和小鼠微生物监测结果分析：以复旦大学实验动物科学部为例[J]. 实验动物与比较医学, 2023, 43(4): 347-354.

Ying HUANG, Siyu WEI, Li CAI, Sujing QIANG, Dongting LI, Yuqiang DING. 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.

图/表 4

参考文献 17

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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)
大鼠细小病毒Rat parvovirus^*	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)

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)
大鼠细小病毒Rat parvovirus^*	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)

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
项目 Pathogens	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.

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
项目 Pathogens	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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