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Definition of Microbiological Status of Rats and Mice / The Need for Methods of Defining Flora / International Standards for Terminology

Kazuaki Mannen

Associate Professor, Laboratory Animal Research Center

Oita Medical University

Japan

Current Status of Microbiological Quality of Laboratory Animals in University Animal Centers in Japan

The organization of university animal centers in Japan is shown in Figure 1. The national university animal centers consist of 53 facilities. The Division of Science and International Affairs Bureau of the Ministry of Education, Science, Sports, and Culture, of which Mr. Ota is Director, is closely linked with all of the animal centers.

In Japan, the national university medical schools and national institutions are members of the Association of Laboratory Animal Facilities of the National University in Japan, hereafter referred to as University Facilities Japan (UFJ). We occasionally encounter subtle differences between the required microbiological inspection of animals being transported among the national, public, and private colleges and other, atypical locations such as other academic institutions or nonapproved vendors and researchers. This type of problem also exists with international transportation. Because specific pathogen-free (SPF) animals are under strict microbiological control, it is technically not necessary to consider pathogenic contamination during their transportation to and from UFJ locations. However, we have found during quarantine inspection that gene-manipulated animals, such as transgenic and knockout mice and rats, have been contaminated by some microorganisms. For this reason, UFJ has established a Working Biohazard Committee (of which I am a member) to formulate guidelines for the microbiological quarantine inspection of mice and rats.



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Definition of Microbiological Status of Rats and Mice / The Need for Methods of Defining Flora / International Standards for Terminology Kazuaki Mannen Associate Professor, Laboratory Animal Research Center Oita Medical University Japan Current Status of Microbiological Quality of Laboratory Animals in University Animal Centers in Japan The organization of university animal centers in Japan is shown in Figure 1. The national university animal centers consist of 53 facilities. The Division of Science and International Affairs Bureau of the Ministry of Education, Science, Sports, and Culture, of which Mr. Ota is Director, is closely linked with all of the animal centers. In Japan, the national university medical schools and national institutions are members of the Association of Laboratory Animal Facilities of the National University in Japan, hereafter referred to as University Facilities Japan (UFJ). We occasionally encounter subtle differences between the required microbiological inspection of animals being transported among the national, public, and private colleges and other, atypical locations such as other academic institutions or nonapproved vendors and researchers. This type of problem also exists with international transportation. Because specific pathogen-free (SPF) animals are under strict microbiological control, it is technically not necessary to consider pathogenic contamination during their transportation to and from UFJ locations. However, we have found during quarantine inspection that gene-manipulated animals, such as transgenic and knockout mice and rats, have been contaminated by some microorganisms. For this reason, UFJ has established a Working Biohazard Committee (of which I am a member) to formulate guidelines for the microbiological quarantine inspection of mice and rats.

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Figure 1 Organizational chart of animal centers in Japanese universities. Importance of Identifying Contamination Forty-nine of the 53 facilities at national universities and centers participated in the survey (Table 1). Of the 49 facilities, 47 (96%) had gene-manipulated mice from domestic sources. A total of 28 facilities (57%) had mice from international sources, mainly from the United States and in some cases from Great Britain, Switzerland, France, Canada, and Germany. The contaminating microorganisms of the mice are shown in Table 2. Major organisms were mouse hepatitis virus, Pasteurella, Mycoplasma, Syphacia, TABLE 1 Introduction of Transgenic and Knockout Mice (1996–1997) Number of facilities responding: 49 (n=53) Facilities with transgenic and knockout mice: 47 (96%) Domestic introduction: 47 (96%) International introduction: 28 (57%) U.S.A. Great Britain American Red Cross Holland Laboratory Mammalian Genetic Unit Charles River Lab Medical Research Council Chrysalis DNX Transgenic Sciences   Harvard University Switzerland Jackson Lab CIBA NCI   NIH France McLaughlin Research Institute Institut Gustave Roussy Northwestern University   North Carolina University Canada Stanford University Ontario Cancer Institute University of Missouri College of Vet Med   University of California Germany   Heidelberg Universitat

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TABLE 2 Contamination at the Introduction of Transgenic and Knockout Mice (1995–1997) Domestic Introduction International Introduction Mouse hepatitis virus (11) Pasteurella spp (12) Pasteurella pneumotropica (10) Trichomonas spp (9) Mycoplasma pulmonis (5) Pneumocystis carini (8) Syphacia spp (5) Mouse hepatitis virus (5) Pseudomonas aeruginosa (4) Duodenum & Cecum for protozoa (4) Sendai virus (3) Helicobacter spp (4) Staphylococcus aureus (2) Mouse poliovirus (GDVII) (3) Trichomonas spp (2) Proteus spp (3) Aspiculuris tetraptera (1) Actinobacillus spp (2) Bordetella bronchiseptica (1) Mouse rotavirus (EDIMV) (2) Corynebacterium Kutscheri (1) Klebsiella spp (2) Hanta virus (HFRS) (1) Mouse parvovirus (2) Octomitus pulcher (1) Pseudomonas aeruginosa (2) Pneumocystis carinii (1) Staphylococcus aureus (2) Polyplax spinulosa (1) Syphacia obvelata (2) Salmonella spp (1) Cecal amebiasis (1) Streptococcus zooepidemics (1) Entamoeba muris (1) Myobia musculi (1) Myobia musculi (1)   Myocoptes musculinus (1)   Streptococcus spp—hemolytic (1)   Theiler's virus (1)   Carbacillus (1) Figure 2 Contamination at the introduction of TG/KO mice between 1995 and 1997.

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Pseudomonas, Sendai virus, and so forth from domestic introduction. In contrast, the spectrum of contaminating organisms resulting from international transportation differed from domestic transportation. As shown in Figure 2, almost all (87 to 92%) of the contaminated mice were accepted conditionally. (Surprisingly, 2% of domestic and 5% of internationally transported mice that were contaminated were accepted unconditionally.) Of the 137 domestic cases, 17% were cleaned up after acceptance; 19% were confined in special rooms; and 63% were not specified. Of the 103 international cases, 61 % were confined in special rooms—roughly three times more than in domestically contaminated mice. Although the reason for this difference is unclear, it may reflect the facility administrators' belief that international contamination is a more serious problem.