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International Committee on Systematic Bacteriology Subcommittee on the Taxonomy of Leptospira Minutes of the Meetings, 13 and 15 September 1990, Osaka, Japan

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International Committee on Systematic Bacteriology Subcommittee on the Taxonomy of Leptospira Minutes of the Meetings, 13 and 15 September 1990, Osaka, Japan INTERNATIONAL JOURNAL OF SYSTEMATICBACTERIOLOGY, Apr. 1992, p. 330-334 Vol. 42, No. 2 0020-7713/92/020330-05$02.00/0 Copyright 0 1992, International Union of Microbiological Societies International Committee on Systematic Bacteriology Subcommittee on the Taxonomy of Leptospira Minutes of the Meetings, 13 and 15 September 1990, Osaka, Japan Minute 1. Call to order. The meeting was called to order reference laboratories maintain a full collection of serovars, by the Chairman, E. Kmety, at 1300 on 13 September 1990. and (v) production and distribution of monoclonal antibod- The opening consisted of a welcoming introduction. Follow- ies. ing this there were additional open sessions at 1800 on 13 Minute 9. General aspects of taxonomy. E. Kmety pre- September and at 0900 on 15 September. A closed session sented a paper on general aspects of taxonomy. He spoke of was held at 1300 on 15 September. early attempts at a phylogenetic classification and also of Minute 2. Record of attendance. The members present problems in the use of DNA base composition for compari- were E. Kmety (Chairman), R. Yanagawa (Vice-chairman), son purposes. Following this he discussed problems with N. Stallman (Secretary), M. Cinco, H. Dikken, W. Ellis, S. using DNA hybridization. It was stated that some microbi- Faine, R. Johnson, R. Marshall, and W. Terpstra. In addi- ologists have considered the possibility of having two clas- tion to members of the subcommittee, the following individ- sification systems, one phylogenetic and the other practical. uals attended one or more of the open sessions: B. Adler, A. Comments were then made on a report submitted by the Alexander, B. Cacciapuoti, W. Chang, S. Coghlan, T. Fu- International Committee on Systematic Bacteriology Ad jikura, Y. Kobayashi, H. Korver, T. Masuzawa, P. Perolat, Hoc Committee on Reconciliation of Approaches to Bacte- A. Schonberg, Y. Yanagihara, and S. Yamamoto. rial Systematics in 1987. In this report it was stated that at Minute 3. Apologies for absence. Apologies for absence present the species is the only taxonomic unit that can be were received from G. Baranton, Dai Bao-min, D. Brenner, defined in phylogenetic terms. According to the phylogenetic Nie Di-kai, C. Everard, K. Hovind-Hougen, A. Kaufmann, definition, a species generally includes strains that exhibit J. Mazzonelli, and A. Thiermann. approximately 70% or greater DNA-DNA relatedness. Minute 4. Mark of respect. The subcommittee recorded There is some evidence that the taxon subspecies is phylo- with deepest regret the death of a member, Y. G. Cher- genetically valid, and further guidelines are needed for nukha, and the death of H. Willinger, who was a collabora- definition of this taxonomic unit. It was also stated that tor. nomenclature should reflect genomic relationships to the Minute 5. Report on the activities of the subcommittee. A greatest extent possible. The overall concern of members of report of the subcommittee activities for the period from the Ad Hoc Committee on Reconciliation of Approaches to 1987 to 1990 was read and accepted. Bacterial Systematics was that any phylogenetically based Minute 6. Minutes of theprevious meeting. The minutes taxonomic schemes must show phenotypic consistency. of the meeting held in Manchester, England, on 5 and 6 Minute 10. Present status of Leptospira parva. W. Ellis September 1986, which were circulated on 10 November presented a paper on the known features of Leptospiraparva 1986, were accepted as a true record of the meeting. as described by Hovind-Hougen et al. in 1981, with subse- Minute 7. Matters arising from the minutes of the previ- quent additional observations by Yasuda et al. in 1987 and ous meeting. (i) Standardization of methods used for the by Saito et al. in 1987. A lengthy discussion followed on the identification of serovars. A discussion was held on heat- differences between Leptospira parva and the genera labile antigens, and it was agreed that the current method Leptospira and Leptonema. The members of the subcom- used for classification, in which whole antigen is used, mittee agreed that a number of differences exist and that a should continue to be used. proposal for a new genus, Tumeria, the type strain of which (ii) Proposed new classification based on DNA homology. is Tumeria parva H, should be published in the International An error in spelling was detected; Leptospira santarosan' Journal of Systematic Bacteriology. should be corrected to Leptospira santarosai. Minute 11. Use of Ambis as a taxonomic tool. W. Ellis Minute 8. WHO activities in international cooperation on presented a paper on the use of Ambis as a taxonomic tool. leptospirosis control and research. T. Fujikura presented a The Ambis system was described as a numerical classifica- paper on the activities being developed by the World Health tion system that is based on a comparison of polypeptide Organization (WHO) for the prevention and control of banding patterns and levels of DNA homology. It provides leptospirosis and the promotion of research in collaboration methods for recording, comparing, and manipulating sodium with the scientific community, member states, research dodecyl sulfate-polyacrylamide gel electrophoresis separa- institutions, and other international organizations. The ac- tions. The scan patterns can be compared with patterns of tivities were discussed under the following headings: (i) other known and unknown organisms which are held in a monitoring and surveillance of leptospirosis; (ii) develop- data bank. The short scan time allows rapid identification. ment of a world network of WHO collaborating centers and The system is very costly and not cost effective at this stage. related international reference institutions; (iii) international It was stated that it is too early to comment on the use of this collaborative research on leptospirosis; (iv) WHO meetings system as an epidemiological tool. A comparison of serovar and training courses and elaboration of guidelines for diag- hardjo bovis strains obtained from various areas of the world nosis and control of leptospirosis; and (v) perspectives of was described. international cooperation. Following the presentation a dis- Minute 12. Genetic characterization of pathogenic cussion was held on (i) vaccine production, (ii) assistance Leptospira species by DNA hybridization. R. Marshall pre- from the WHO in translating Chinese publications into sented a paper on genetic characterization of pathogenic English, (iii) designation of a reference laboratory in Africa, Leptospira species by DNA hybridization. He described a (iv) problems in diagnostic procedures because only a few quantitative slot blot hybridization method that he used to 330 VOL.42, 1992 MINUTES 331 determine the levels of DNA relatedness among 66 patho- alternative classification is considered, a system based on genic Leptospira serovars; in this study 16 strains were used phylogenetic relationships would be the most logical. Also, as sources of labeled reference DNA, and 57 of the serovars restriction endonuclease analysis is not very suitable for were placed into six DNA homology groups. The results classification. In the long term, investigation into the com- were in general agreement with the results of other workers position of DNA as a basis for determining phylogenetic who used different DNA homology methods. As a result of relationships might lead to a natural classification system. It differences, the new species Leptospira kirschneri was pro- was concluded that pending the possible development of a posed and into it were placed serovars cynopteri, grippoty- new classification system based on phylogenetic relation- phosa, kabura, tsaratsovo, butembo, kambale, ramisi, dania, ships, it is practical to maintain the system based on cross- and bulgarica. It was suggested that after serogroup Hard- agglutinin absorption results. However, data for new strains joprajitno strains are examined, the terms Hardjoprajitno should be complemented by restriction endonuclease analy- and Bovis should be used. A discussion was held, and it sis patterns. was agreed that confusion may occur if this change were Minute 16. Thermolabile antigens. E. Kmety commented made. on thermolabile antigens. He stated that sometimes these Minute 13. Leptospira biflexa: remarks on a very hetero- antigens are present and sometimes they are not. He also geneous species. M. Cinco presented a paper on Leptospira stated that the Vi antigen is present in the pathogenic biflexa, a very heterogeneous species. Using various meth- leptospires and is related to virulence. Thermolabile antigens ods, he described the genetic diversity of strains in the were discovered in 1972 and have been shown to be present Leptospira biflaa group and stated that the diversity in this in serogroups Icterohaemorrhagiae and Pomona. The fact species is greater than that in Leptospira interrogans. Of 65 that this finding has been neglected in the past has led to known saprophytic strains, 24 do not fit into groups. These discrepant results. It was stated that thermolabile antigens 24 strains are individual strains, and each represents one could be lost over the years. The presence of thermolabile serovar and one serogroup. It was stated that on the basis of antigens is not important taxonomically. At present we use a observations Leptospira biflexa is a very heterogeneous two-antigen system based on thermolabile antigens. Antisera taxon which contains strains that do not exhibit sufficient are prepared from whole antigen, which is a mixture of DNA relatedness to accept them as members of one species. thermolabile and thermostable antigens. It is possible that It would be more appropriate to use the term nonpathogenic the recommendation relating to thermostable antigens may leptospires instead of the binomial Leptospira biflaa. If we have to be changed. assume that a genetic criterion for taxonomy should be used, Minute 17. Hybridization. R. Marshall presented a paper further studies on the serovars will be needed, and individual on hybridization and its impact on classification. He outlined species names may be given to each genetic group. a method in which single-stranded DNA from one organism Minute 14.
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