1887

Abstract

A group of seven sucrose-negative strains (referred to as group Au) isolated from the internal organs of septicaemic farmed frogs () in Thailand was subjected to a polyphasic taxonomic study including fluorescent amplified fragment length polymorphism (FAFLP) and ERIC-PCR fingerprinting, 16S rDNA sequencing, microplate DNA–DNA hybridizations and extensive phenotypic characterization. Comparison of FAFLP and ERIC-PCR fingerprints indicated that the group Au isolates belonged to the species DNA hybridization group (HG) 1 in which they represent a genotypic subgroup closely affiliated to subsp. and subsp. . One representative of the Au group exhibited ⩾99·0 % 16S rDNA sequence similarity with the type strains of the two subspecies. DNA–DNA hybridization with type and reference strains of all known taxa revealed that the Au group represented a homogeneous taxon that exhibited the highest relatedness with members of the two subspecies, ranging from 75 to 93 %. Phenotypic characterization on the basis of 152 features further revealed that the Au group isolates differed from subsp. or subsp. in a total of 13 biochemical properties. Of these, assimilation of -glycine and isobutyrate as sole carbon source, acid production from salicin and -sucrose, and aesculin hydrolysis were of diagnostic value. From the results of this study, it can be concluded that the frog isolates of the Au group represent a new subspecies of , for which the name subsp. subsp. nov. is proposed. Its type strain is Au-1D12 (=LMG 19707=CCUG 46211).

Loading

Article metrics loading...

/content/journal/ijsem/10.1099/ijs.0.02357-0
2003-05-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/ijsem/53/3/ijs530885.html?itemId=/content/journal/ijsem/10.1099/ijs.0.02357-0&mimeType=html&fmt=ahah

References

  1. Abbott S. L., Cheung W. K. W., Kroske-Bystrom S., Malekzadeh T., Janda J. M. 1992; Identification of Aeromonas strains to the genospecies level in the clinical laboratory. J Clin Microbiol 30:1262–1266
    [Google Scholar]
  2. Altwegg M., Lüthy-Hottenstein J. 1991; Methods for the identification of DNA hybridization groups in the genus Aeromonas . Experientia 47:403–406
    [Google Scholar]
  3. Bauer A. W., Kirby W. M. M., Sherris J. C., Turck M. 1966; Antibiotic susceptibility testing by a standardized single disc method. Am J Clin Pathol 45:493–496
    [Google Scholar]
  4. Carnahan A., Fanning G. R., Joseph S. W. 1991; Aeromonas jandaei (formerly genospecies DNA group 9 A. sobria ), a new sucrose-negative species isolated from clinical specimens. J Clin Microbiol 29:560–564
    [Google Scholar]
  5. Ezaki T., Hashimoto Y., Yabuuchi E. 1989; Fluorometric deoxyribonucleic acid - deoxyribonucleic acid hybridization in microdilution wells as an alternative to membrane filter hybridization in which radioisotopes are used to determine genetic relatedness among bacterial strains. Int J Syst Bacteriol 39:224–229 [CrossRef]
    [Google Scholar]
  6. Goris J., Suzuki K.-I., De Vos P., Nakase T., Kersters K. 1998; Evaluation of a microplate DNA-DNA hybridization method compared with the initial renaturation method. Can J Microbiol 44:1–7 [CrossRef]
    [Google Scholar]
  7. Gosling P. J. 1996; Aeromonas species in disease of animals. In The Genus Aeromonas pp 175–196Edited by Austin B., Altwegg M., Gosling P. J., Joseph S. New York: Wiley;
    [Google Scholar]
  8. Hänninen M.-L. 1994; Phenotypic characteristics of the three hybridization groups of Aeromonas hydrophila complex isolated from different sources. J Appl Bacteriol 76:455–462 [CrossRef]
    [Google Scholar]
  9. Hickman-Brenner F. W., Fanning G. R., Arduino M. J., Brenner D. J., Farmer J. J. III 1988; Aeromonas schubertii , a new mannitol-negative species found in human clinical specimens. J Clin Microbiol 26:1561–1564
    [Google Scholar]
  10. Hirono I., Aoki T. 1993; Cloning and characterisation of three haemolysin genes from Aeromonas salmonicida . Microb Pathog 15:269–282 [CrossRef]
    [Google Scholar]
  11. Huys G., Swings J. 1999; Evaluation of a fluorescent amplified fragment length polymorphism (FAFLP) methodology for the genotypic discrimination of Aeromonas taxa. FEMS Microbiol Lett 177:83–92 [CrossRef]
    [Google Scholar]
  12. Huys G., Kämpfer P., Altwegg M., Coopman R., Janssen P., Gillis M., Kersters K. 1997a; Inclusion of Aeromonas DNA hybridization group 11 in Aeromonas encheleia , and extended descriptions of the species Aeromonas eucrenophila and A. encheleia. Int J Syst Bacteriol 471157–1164 [CrossRef]
    [Google Scholar]
  13. Huys G., Kämpfer P., Altwegg M.7 other authors 1997b; Aeromonas popoffii sp. nov., a mesophilic bacterium isolated from drinking water production plants and reservoirs. Int J Syst Bacteriol 47:1165–1171 [CrossRef]
    [Google Scholar]
  14. Huys G., Gevers D., Temmerman R.8 other authors 2001; Comparison of the antimicrobial tolerance of oxytetracycline-resistant heterotrophic bacteria isolated from hospital sewage and freshwater fishfarm water in Belgium. Syst Appl Microbiol 24:122–130 [CrossRef]
    [Google Scholar]
  15. Huys G., Kämpfer P., Albert M. J., Kühn I., Denys R., Swings J. 2002; Aeromonas hydrophila subsp. dhakensis subsp. nov., isolated from children with diarrhoea in Bangladesh, and extended description of Aeromonas hydrophila subsp. hydrophila , (Chester 1901). Stanier 1943 (Approved Lists 1980). Int J Syst Evol Microbiol 52705–712 [CrossRef]
    [Google Scholar]
  16. Janda J. M., Abbott S. 1996; Human pathogens. In The Genus Aeromonas pp 151–174Edited by Austin B., Altwegg M., Gosling P. J., Joseph S. New York: Wiley;
    [Google Scholar]
  17. Joseph S. W., Carnahan A. 1994; The isolation, identification, and systematics of the motile Aeromonas species. Annu Rev Fish Dis 4:315–343 [CrossRef]
    [Google Scholar]
  18. Kämpfer P., Altwegg M. 1992; Numerical classification and identification of Aeromonas genospecies. J Appl Bacteriol 72:341–351 [CrossRef]
    [Google Scholar]
  19. Marmur J. 1961; A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol 3:208–218 [CrossRef]
    [Google Scholar]
  20. Martinez-Murcia A. J. 1999; Phylogenetic positions of Aeromonas encheleia , Aeromonas popoffii , Aeromonas DNA hybridization group 11 and Aeromonas Group 501. Int J Syst Bacteriol 49:1403–1408 [CrossRef]
    [Google Scholar]
  21. Martinez-Murcia A. J., Esteve C., Garay E., Collins M. D. 1992; Aeromonas allosaccharophila sp. nov., a new mesophilic member of the genus Aeromonas . FEMS Microbiol Lett 91:199–206 [CrossRef]
    [Google Scholar]
  22. National Committee for Clinical Laboratory Standards (NCCLS) 1993 Performance Standards for Antimicrobial Disk Susceptibility Tests , 5th edn. Approved standard M2-A5 Vilanove, PA: National Committee for Clinical Laboratory Standards;
    [Google Scholar]
  23. Pasquale V., Baloda S. B., Dumontet S., Krovacek K. 1994; An outbreak of Aeromonas hydrophila infection in turtles ( Pseudemis scripta . Appl Environ Microbiol 60:1678–1680
    [Google Scholar]
  24. Pavan M. E., Abbott S. L., Zorzopulos J., Janda J. M. 2000; Aeromonas salmonicida subsp. pectinolytica subsp. nov., a new pectinase-positive subspecies isolated from a heavily polluted river. Int J Syst Evol Microbiol 50:1119–1124 [CrossRef]
    [Google Scholar]
  25. Pearson M. D., Colquhoun D., Somsiri T., Inglis V. 1997; Biochemical characterisation and RAPD analysis of an Aeromonas species isolated from septicaemic Rana rugulosa (Weigmann) cultured in Thailand. In Diseases in Asian Aquaculture III pp 9–14Edited by Flegel T. W., McRae I. H. Manila: Fish Health Section, Asian Fisheries Society;
    [Google Scholar]
  26. Pearson M. D., Hirono I., Aoki T., Miranda R., Inglis V. 2000; Virulence properties of motile aeromonads isolated from farmed frogs Rana tigerina and R. rugulosa . Dis Aquat Org 40:185–193 [CrossRef]
    [Google Scholar]
  27. Pitcher D. G., Saunders N. A., Owen R. J. 1989; Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Lett Appl Microbiol 8:151–156 [CrossRef]
    [Google Scholar]
  28. Rigney M. M., Zilinsky J. W., Rouf M. A. 1978; Pathogenicity of Aeromonas hydrophila in red leg disease in frogs. Curr Microbiol 1:175–179 [CrossRef]
    [Google Scholar]
  29. Traub W. H., Geipel U., Leonhard B. 1998; Antibiotic susceptibility testing (agar disk diffusion and agar dilution) of clinical isolates of Enterococcus faecalis and E. faecium : comparison of Mueller-Hinton, Iso-sensitest, and Wilkins-Chalgren agar media. Chemotherapy 44:217–229 [CrossRef]
    [Google Scholar]
  30. Versalovic J., Koeuth T., Lupski J. R. 1991; Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res 19:6823–6831 [CrossRef]
    [Google Scholar]
  31. Versalovic J., Schneider M., De Bruijn F. J., Lupski J. R. 1994; Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods Mol Cell Biol 5:25–40
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/ijsem/10.1099/ijs.0.02357-0
Loading
/content/journal/ijsem/10.1099/ijs.0.02357-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error