J. Gen. Appl. Microbiol., 43, 49-59 (1997)

Polyamine distribution patterns within the families , , , and , and related genera of the gamma subclass of the

Koei Hamana

College of Medical Care and Technology, Gunma University, Maebashi 371, Japan

(Received June 12, 1996; Accepted December 19,1996)

Polyamines of the four families and the five related genera within the gamma subclass of the class Proteobacteria were analyzed by HPLC with the objective of developing a chemotaxonomic system. The production of putrescine, diaminopropane, cadaverine, and agmatine are not exactly correlated to the phylogenetic genospecies within 36 strains of the genus (the family Aeromon- adaceae) lacking in triamines. The occurrence of norspermidine was limited but not ubiquitous within the family Vibrionaceae, including 20 strains of , Listonella, Photobacterium, and Salini- vibrio. Spermidine was not substituted for the absence of norspermidine in the family. Agmatine was detected only in Photobacterium. Salinivibrio and some strains of Vibrio were devoid of polyamines. Vibrio ("Moritella") marinus contained cadaverine. Within the family Pasteurellaceae, contained cadaverine only and contained no polyamine. , , and , belonging to the family Halomonadaceae, ubiquitously con- tained spermidine and sporadically cadaverine and agmatine. Shewanella contained putrescine and cadaverine; Alteromonas macleodii, putrescine, 2-hydroxyputrescine, cadaverine, 2-hydroxysper- midine, and spermidine; Pseudoalteromonas, putrescine, cadaverine, and spermidine; Marinobacter, spermidine; and Marinomonas, putrescine and spermidine. Their polyamine profiles serve as a chemotaxonomic marker within the gamma subclass.

Key Words Aeromonadaceae; Halomonadaceae; norspermidine; Pasteurellaceae; polyamine; Pro- teobacteria; Vibrionaceae

Studies on cellular polyamine distribution have al- Aeromonas was placed in the family Aeromon- ready provided some valuable chemotaxonomic infor- adaceae as the only genus (Colwell et al., 1986). The mation within the class Proteobacteria, and are usu- members of the genus Aeromonas were divided into ally characteristic for higher taxonomic rank such as at least 16 genospecies by DNA homology (Abbott et for the many members of the alpha subclass of the al., 1992; Ali et al., 1996; Altwegg et al., 1990; Carna- Proteobacteria, which feature homospermidine, or for han et al., 1991a, 1991b, 1991c; Collins et al., 1993; the organisms belonging to the beta subclass, which Esteve et al., 1995a, 1995b; Huys et al., 1994; Janda, feature 2-hydroxyputrescine (Auling et al., 1991; 1991; Joseph et al., 1991; Lucchini and Altwegg, Busse and Auling, 1988; Hamana and Matsuzaki, 1992; Martinez-Murcia et al., 1992a, 1992b; Sneath, 1992a, 1993; Yang et al., 1993). The four families, 1993), however, taxonomic conclusions are not com- Aeromonadaceae, Vibrionaceae, Pasteurellaceae, plete. We analyzed polyamines as the phenotypic and , are major members of the marker of 36 strains of Aeromonas, including 15 gamma subclass of the Proteobacteria and comprise genospecies, for chemotaxonomic interest. a cluster in the subclass. In the four families, determi- The phylogenetic classification of the genera Vibrio, nation of the polyamines of Enterobacteriaceae, in Photobacterium, and Listonella belonging to Vibri- which diaminopropane and acetyispermidine are onaceae is conflicting (Holmes, 1992; Kita-Tsukamoto widely spread, has been published (Hamana,1996). et al., 1993; Ruimy et al., 1994; Smith et al., 1991). Norspermidine has been found as a major polyamine

Address reprint requests to: Dr. Koei Hamana, College of Medical in many Vibrio, Listonella, and Photobacterium Care and Technology, Gunma university, 3-39-15 Showa-machi, , but some norspermidine-absent strains were Maebashi 371, Japan. reported within these three genera (Yamamoto et al., 50 HAMANA Vol. 43

1983, 1991). In this study, the polyamines of some contained 21 amino acids, 17 vitamins, 10 nu- new species of the above genera (Ishimaru et al., cleotides, sodium acetate, glucose, NaCI, KCI, CaCl2, 1995, 1996; Okuzumi et al., 1994), as well as the MgSO4, Na2HPO4, and Fe(NO3)3, but not ornithine, di- newly proposed genera Salinivibrio (Mellado et al., aminobutyric acid or polyamines. Putrescine, cadaver- 1996) and "Moritella" (Gauthier et al., 1995a) in Vibri- me, agmatine, spermidine, and spermine were de- onaceae, were analyzed in comparison with the tected in GAM, NB, BHIB, PY SW, and MB. The Acti- polyamine types found in other related marine AI- nobacillus and Haemophilus species were cultivated teromonas, Shewanella, Marinomonas, and Oceano- at 37°C and Shewanella benthica at 2°C, whereas the spirillum species, newly isolated Marinobacter hydro- others were grown at 20-30°C. Actinobacillus actino- carbonoclasticus (Gauthier et al., 1992) and the newly mycetecomitans was grown in the presence of 5% validated genus Pseudoalteromonas (Gauthier et al., CO2. 1995a) within the gamma subclass of the Proteobac- The pellets of cells harvested at the late-stationary teria. phase were homogenized in 0.5M perchloric acid The polyamines of some members of the genera (HCIO4) at 2°C. Polyamines were extracted into HCIO4 , Haemophilus, and Actinobacillus (Gau- and analyzed by high-performance liquid chromatog- thier et al., 1995b), which belong to the family Pas- raphy on a L600 high-speed liquid chromatograph teurellaceae in the gamma subclass of the class Pro- (Hitachi Co., Tokyo, Japan) as described previously teobacteria, were also determined. (Hamana and Matsuzaki, 1992b,1993). Phylogenetic analyses of the halophilic proteobacte- ria belonging to the genera Halomonas, Deleya, and Results and Discussion and two halophilic organisms, Volcaniella eurihalina and Paracoccus halodenitrificans, showed The major polyamines detected in the four families that these proteobacteria constitute another major (Aeromonadaceae, Vibrionaceae, Pasteurellaceae, family of the gamma subclass, Halomonadaceae and Halomonadaceae) and six related genera of ma- (Dobson et al., 1993; Mellado et al., 1995; Miller et al., rine (Shewanella, Pseudoalteromonas, AI- 1994). Unification of these genera into a single genus teromonas, Marinobacter, Marinomonas, and Oceano- (Halomonas), and placement of Chromohalobacter spirillum) are summarized into a schematic phyloge- marisomortui, and the newly isolated Zymobacter pal- netic tree in Fig. 1. The details of the polyamine com- mae (Okamoto et al., 1993) in the family, were pro- positions found in the four families and related genera posed (Dobson and Franzmann, 1996; Mellado et al., are shown separately in five tables, Tables 1-5, re- 1995). Although the major polyamine of some species spectively. of Halomonas and Deleya was spermidine in the pre- vious publication (Hamana and Matsuzaki, 1993), the Aeromonadaceae ( Table 1) polyamines of the new members of Halomonadaceae Ubiquitous distribution of putrescine and the ab- were analyzed in this study. sence of triamines such as spermidine and norspermi- dine were found in all strains of the Aeromonas Materials and Methods species tested in this study, as well as a report for some Aeromonas species by Yamamoto et al. (1991). Organisms were aerobically cultured in a synthetic When Aeromonas was grown in NB, the minor medium, 199 medium (199), pH 7.2 (Nissui Pharma- polyamines detected in the cultures was excluded (as ceutical Co., Tokyo, Japan); GAM medium (GAM), pH a contaminant) from the net cellular polyamine com- 7.0 (Nissui Pharmaceutical Co.); nutrient broth (NB), ponents. Variations in the distribution profiles of di- pH 7.0 (Nissui Pharmaceutical Co.); and brain heart aminopropane, cadaverine, histamine, and agmatine infusion broth (BHIB) supplemented with 5% Fildes were observed within the same species (genotype): Enrichment (Difco Lab., Detroit, MI, U.S.A.). PY putrescine type and putrescine-cadaverine type in medium (1% polypeptone, 0.2% yeast extract, and A, allosaccharophila (HG 14) and A. salmonicida (HG 0.1% MgSO4) dissolved in 75% seawater (PY SW), 3); and putrescine-cadaverine type and putrescine- the 199 medium dissolved in 75% seawater (199- cadaverine-agmatine type in A. schubertii (HG 12). SW), IFO 306 medium (List of Cultures, 10th ed., Diaminopropane and histamine were sporadically 1996, IFO), Marine broth 2216 (MB) (Difco Lab.), and spread in some genospecies. These findings indicate IAM B-15 and IAM B-84 media (Catalogue of Strains, that the appropriate decarboxylase activities to pro- 1st ed., 1993, IFO) were used for halophiles. The 199- duce diaminopropane, cadaverine, histamine, and ag- SW and PY SW supplemented with 10 mM L-2,4-di- matine are not exactly correlated to the phylogenetic aminobutyric acid, L-ornithine, 1,3-diaminopropane genospecies within Aeromonas, and that these amine and/or putrescine were also used. The 199 medium profiles are not useful as a phenotypic marker within 1997 Polyamines of Aeromonadaceae, Vibrionaceae, Pasteurellaceae, and Halomonadaceae 51

me was found but norspermidine was not detected. In this study, the absence of norspermidine in this organ- ism is clear. This polyamine profile for V. marinus ATCC 15381T supports the phylogenetic finding that V. marinus ATCC 15381T should be placed in a new separate genus,"Moritella," as "M. marinus" (Gauthier et al.,1995a). The newly isolated species of Vibrio, V. orientalis, V. penaeicida, V. ordalli, and "V. algoinfesta," con- tained norspermidine. Spermidine was detected in the former three organisms grown in PY-SW containing spermidine, whereas the last Vibrio species lacked in spermidine when it was cultivated in PY SW. A new species of Photobacterium, P, histaminum, grown in 199 contained histamine and agmatine as the major polyamines, and was devoid of norspermidine and spermidine. Although putrescine and spermidine ap- peared in this organism when it was grown in PY SW, the synthetic abilities of putrescine, norspermidine, and spermidine from the supplemented ornithine, di- aminopropane, and putrescine were not detected. The media used for the cultures of Vibrio and Photobac- terium sometime affected the cellular spermidine lev- els. The uptake of spermidine into the cells from sper- midine-containing PY SW medium without the net syn- thesis of spermidine is possible. Among the 83 strains of Vibrionaceae, the absence of norspermidine in two strains of each V. costicola, V fischeri, and P. phosphoreum, and no synthesis of tri- amine from the supplemented diaminopropane in their Fig. 1. Major polyamine types on the unrooted phylogenetic cultures, has been suggested by Yamamoto et al. tree of the gamma subclass of the Proteobacteria derived from 16S (1983, 1991). When we cultured some strains of the rRNA sequences (Dobson and Franzmann, 1996; Gauthier et al., three species in two different culture media and rean- 1995a, 1995b; Mellado et al., 1995; Ruimy et al., 1994). alyzed the polyamines, the absence of norspermidine Four families (Aeromonadaceae, Vibrionaceae, Pasteurellaceae, and Halomonadaceae) are shown separately in A-D corresponding in the three species was confirmed. V. costicola, V. to Tables 1-4, respectively. The data of the five genera (Pseudoal- hollisae, and P. damsela are phylogenetically located teromonas, Shewanella, Alteromonas, Marinobacter, and Mari- on a branch placed on a side end in Vibrionaceae, nomonas) correspond to Table 5. Polyamines of the genus and V. fischeri and V. logei are on another branch. Oceanospirillum were cited from Hamana et al. (1994). Major Therefore, the two groups are divergent from a cluster polyamines sporadically found within the species (or genus) are shown in parentheses. Abbreviations for polyamiries and genus comprising other authentic Vibrio species (Kita- names are shown in Tables 1-5. -, polyamines were not detected. Tsukamoto et al., 1993; Ruimy et al., 1994) In this study, norspermidine was found in V. hollisae and P. damsela, but not in V costicola. Furthermore, V logei the family (genus). The absence of triamines serve contained norspermidine, while V. fischeri was devoid as a chemotaxonomic property of the family (genus of norspermidine, as previously demonstrated by Ya- within the gamma subclass. mamoto et al. (1983, 1991). P. leignathi and P. angus- tum (Yamamoto et al., 1991) contained norspermidine Vibrionaceae (Table 2) as a major polyamine. Therefore, in five Photobac- The occurrence of cadaverine and norspermidine it terium species, P. phosphoreum and P. histaminum Vibrio marinus ATCC 15381T has been reported b~ were absent in norspermidine. Yamamoto et al. (1991). This organism is phylogeneti Yamamoto et al. (1991) and Kamekura et al. (1986) cally divergent and located aside from the family Vibri have reported that V. costicola ATCC 33508T and onaceae (Kita-Tsukamoto et al., 1993; Ruimy et al. NRCC 37001 grown in a chemically defined minimum 1994). When we reanalyzed the polyamine of this or medium were devoid of any polyamines. The occur- ganism grown in 199-SW and PY SW media, cadaver rence of spermidine in norspermidine-absent V costi- 52 HAMANA Vol. 43

Table 1. Cellular polyamines of Aeromonadaceae (Aeromonas). 1997 Polyamines of Aeromonadaceae, Vibrionaceae, Pasteurellaceae, and Halomonadaceae 53

Table 1. (Continued.)

Tahle 2_ Cellular nnlvamines in Vihrionaceae (Vibrio. Photobacterium. and Salinivibriol. 54 HAMANA Vol. 43

Table 2. (Continued.)

Table 3. Cellular polyamines in Pasteurellaceae (Actinobacillus and Haemophilus). 1007 pnl~,aminoc of Aormm~nar~ar+cao 1/ihrinnar+oac pacfoi irollar`oac anrJ I-Ialmm~narJar`o~o

Table 4. Cellular polyamines in Halomonadaceae (Halomonas, Chromohalobacter, and Zymobacter).

cola NCIMB 701, V. fischeri ATCC 7744T, and V. fis- new genus Salinivibrio (Mellado et al., 1996). Four cheri strain 61 was reported by Yamamoto et al. Photobacterium species contained agmatine as a (1983), Spermidine was not detected in the type major polyamine component. This is the first finding, strains of V. costicola, V. fischeri, P. histaminum, or a and the occurrence of agmatine serves for the taxon- new Vibrio species, V ichtyoenteri, grown in the omy of this genus. At present, occurrence of norsper- polyamine-free 199 medium supplemented with pu- midine is limited in the family Vibrionaceae in the trescine in this study; indicating the absence of the tri- gamma subclass, however, it is not ubiquitously dis- amine synthesis from putrescine in these organisms. tributed within the family. The two type strains of V. costicola and V. fischeri did not produce diaminopropane or putrescine from the Pasteurellaceae (Table 3) supplemented diaminobutylic acid and ornithine, re- The data of polyamines for some Actinobacillus and spectively. The two strains of P. phosphoreum grown Haemophilus suggest the absence of norspermidine in the synthetic medium were devoid of spermidine, in Pasteurellaceae. Actinobacillus species, which are whereas spermidine was detected in two other strains devoid of any polyamines with 199 medium, contained of P. phosphoreum by Yamamoto et al. (1983, 1991). a significant amount of spermidine and spermine We propose here that spermidine is not synthesized in when the organisms were grown in GAM medium. the type strains of V. costicola, V. fischeri, V. ichtyoen- The uptake of these polyamines into the organisms teri, P. phosphoreum, or P. histaminumn. It is sug- from the medium was suggested. A unique polyamine gested that the organism containing sperridine in the pattern, cadaverine only, was found in Haemophilus absence of norspermidine is not distributed in Vibri- influenzae. Spermidine and spermine were detected onaceae. Recently, V. costicola was transferred to the in BHIB, however, the two polyamines were not found 56 HAMANA Vol. 43

Table 5. Cellular polyamines in Shewanella, Pseudoalteromonas, Alteromonas, Marinobacter, and Marinomonas.

in the cells grown in BHIB. This organism grew in nei- spermidine as a major polyamine. The previous datum ther GAM nor 199. The transfer of A. actinomycetem- for H. halomophila (Hamana and Matsuzaki, 1993) comitans into the genus Haemophilus is not supported should be corrected. A new Halomonas species, H. by the polyamine profiles as a chemotaxonomic meridiana, contained spermidine, and a new Deleya marker. species, D. salina (recently, Halomonas salina) con- tained spermidine and cadaverine. Cadaverine was Halomonadaceae ( Table 4) sporadically detected within Halomonadaceae, how- Since some of the cultures of Halomonas in our pre- ever, it is known that the cadaverine level is sensitive vious study were unsuccessful (Hamana and Mat- to culturing conditions and medium composition suzaki, 1993), we reanalyzed the polyamines of the (Hamana and Matsuzaki, 1992a). A phylogenetic halomonads grown in several media in this study. The analysis of Chromohalobacter marismortui, Volcaniella halophiles belonging to the genus Halomonas and De- eurihalina (recently, Halomonas eurihalina), and Para- leya (recently, Halomonas) ubiquitously contained coccus halodenitrificans (recently, Halomonas halo- 1997 Pol y amines of Aeromonadaceae, Vibrionacea e, P asteure IIaceae, an d H a lomona d aceae 57 denitrificans) indicated that these three organisms or higher taxon levels within the gamma subclass. should be located in the family Halomonadaceae However, Photobacterium species ubiquitously con- (Dobson et al., 1993; Dobson and Franzmann, 1996; taining agmatine lack in norspermidine and Salinivib- Mellado et al., 1995; Miller et al., 1994). We have rio, and some Vibrio species lack any polyamine syn- demonstrated that the polyamine profiles of P. halo- thetic ability, indicating that polyamine profiles serve denitrificans differ from the polyamine pattern of other for the chemotaxonomic classification for the genus authentic Paracoccus species (Hamana and Mat- level within Vibrionaceae. The synthesis of diamines suzaki, 1992b). C. marismortui as well as H. euri- such as diaminopropane, putrescine, cadaverine, and halina and H. halodenitrificans contained spermidine agmatine are specific for genus or species levels as a major polyamine, supporting the phylogenetic re- within each family. sults. However, C. marismortui was not distinguished from Halomonas by its polyamine profile. Z. palmae The author is indebted to Dr. S. Yamamoto of Okayama Univer- sity for helpful suggestions, and to IAM, IFO, JCM, MAFF, and placed in the family contained putrescine„ cadaverine, ATCC for supplying bacterial strains. spermidine, and agmatine. The occurrence of agma- tine was limited in Z palmae within the family. References Shewanella, Pseudoalteromonas, Alterornonas, Mari- Abbott, S.L., Cheung, W.H.W., Kroske-Bystrom, S., Malekzadeh, T., nobacter, and Marinomonas ( Table 5) and Janda, J. M. (1 992) Identification of Aeromonas strains to the genospecies level in the clinical laboratory. J. C/in. Micro- The Shewanella benthica analyzed in this study biol., 30,1262-1266. contained only putrescine. The major polyamine of au- Akagawa-Matsushita, M., Koga, Y., and Yamasato, K. (1993) DNA thentic Shewanella species grown in several media relatedness among nonpigmented species of A/teromonas and was putrescine or putrescine plus cadaverine. The synonymy of Alteromonas ha/op/anktis (ZoBell and Upham Pseudoalteromonas species show putrescine-spermi- 1944) Reichelt and Baumann 1973 and Alteromonas tetraodo- nis Simidu et al. 1990. Int. J. Syst. Bacteriol., 43, 500-503. dine type and cadaverine sporadically distributed in Ali, A., Carnahan, A.M., Altweeg, M., Luthy-Hottenstein, J., and the genus (Hamana and Matsuzaki, 1993). Al- Joseph, SW. (1996) sp. nov. (formerly teromonas macleodii, placed in a separate genus from genospecies DNA group 2 A. hydrophila), a new species iso- Pseudoalteromonas (Akagawa-Matsushita et al., 1993; lated from non-human sources. Med. Microbiol. Lett., 5, 156- Gauthier et al., 1995a; Ivanova et al., 1996), con- 165. Altwegg, M., Steigerwalt, A.G., Altwegg-Bissig, R., Luthy-Hotten- tained two novel polyamines (2-hydroxyputrescine stein, J., and Brenner, D.J. (1990) Biochemical identification of and 2-hydroxyspermidine). As already reported, this is Aeromonas genospecies isolated from humans. J. Clin. Micro- a unique species whithin the former description of the biol., 28, 258-264. genus Alteromonas (Hamana and Matsuzaki, 1993). Auling, G., Busse, H.-J., Pilz, F., Webb, L., Kneifel, H., and Claus, Recently, Gauthier et al. (1995a) proposed the divi- D. (1991) Rapid differentiation, by polyamine analysis, of Xan- thomonas strains from phytopathogenic pseudomonads and sion of the genus Alteromonas into two genera, Al- other members of the class Proteobacteria interacting with teromonas and Pseudoalteromonas. The polyamine plants. Int. J. Syst. Bacterio/., 41, 223-228. profiles of the species presented here strongly support Busse, J. and Auling, G. (1988) Polyamine pattern as a chemotaxo- their proposal from the phenotypic properties. A ma- nomic marker within the Proteobacteria. Syst. App/. Microbiol., rine proteobacterium, Marinobacter hydrocarbonoclas- 11,1-8. Carnahan, A. M., Behram, S., and Joseph, S. W. (1991a) Aerokey ticus, contained spermidine only. Two other marine II: A flexible key for identifying clinical Aeromonas species. J. proteobacteria belonging to the gamma subclass, Clin. Microbiol., 29, 2843-2849. Marinomonas and Oceanospirillum, contained pu- Carnahan, A. M., Chakraborty, T., Fanning, G. R., Verma, D., Ali, A., trescine and spermidine (Hamana and Matsuzaki, Janda, J. M., and Joseph, S. W. (1991b) Aeromonas trota sp. 1993; Hamana et al., 1994). The polyamine profiles of nov., an Ampicillin-susceptible species isolated from clinical specimens. J. Clin. Microbiol., 29,1206-1210. these three marine organisms are identical to that of Carnahan, A. M., Fanning, G. R., and Joseph, S. W. (1991c) Halomonadaceae rather than Vibrionaceae. (formerly genospecies DNA group 9 A. so- The occurrence of norspermidine was limited to the bria), a new sucrose-negative species isolated from clinical family Vibrionaceae within the five families and seven specimens. J. Clin. Microbiol., 29, 560-564. related genera of the gamma subclass of Proteobac- Collins, M.D., Martinez-Murcia, A. J., and Cai, J. (1993) Aeromonas enteropelogenes and Aeromonas ichthiosmia are identical to teria. The absence of triamines such as spermidine Aeromonas trota and , respectively, as re- and norspermidine in Aeromonadaceae and Pas- vealed by small-subunit rRNA sequence analysis. Int. J. Syst. teurellaceae, and the ubiquitous occurrence of sper- Bacteriol., 43, 855-856. midine as the major polyamine in Halomonadaceae Colwell, R. R., MacDonell, M. T., and DeLey, J. (1986) Proposal to and three related genera of Marinobacter, Mari- recognize the family Aeromonadaceae fam. nov. Int. J. Syst. Bacteriol., 36, 473-477. nomonas, and Oceanospirillum, suggest that the Dobson, S. J. and Franzmann, P. D. (1996) Unification of the gen- polyamine distribution pattern is correlated in families era Deleya (Baumann et al. 1983), Ha/omonas (Vreeland et al. 58 HAMANA Vol. 43

1980), and Ha/ovibrio (Fendrich 1988) and the species Para- Yoshikawa, S. (1996) Characterization and identification of ma- coccus halodenitrificans (Robinson and Gibbons 1952) into a rine Alteromonas nigrifaciens strains and emendation of the de- single genus, Halomonas, and placement of the genus Zy- scription. Int. J. Syst. Bacteriol., 46, 223-228. mobacter in the family Halomonadaceae. Int. J. Syst. BacterioL, Janda, M. J. (1 991) Recent advances in the study of the , 46, 550-558. pathogenicity, and infections syndromes associated with genus Dobson, S. J., McMeekin, T. A., and Franzmann, P. D. (1993) Phy- Aeromonas. Clin. Microbiol. Rev., 4, 397-410. logenetic relationships between some members of the genera Joseph, S. W., Carnahan, A. M., Brayton, P. R., Fanning, G. R., AI- Deleya, Halomonas, and Halovibrio. Int. J. Syst. Bacteriol., 43, mazan, R., Drabick, C., Trudo, E.W., Jr., and Colwell, R. R. 665-673. (1991) Aeromonas jandaei and Aeromonas veronii dual infec- Esteve, C., Gutierrez, M. C., and Ventosa, A. (1995a) DNA related- tion of a human wound following aquatic exposure. J. C/in. Mi- ness among Aeromonas allosaccharophila strains and DNA hy- crobiol., 29, 565-569. bridization groups of the genus Aeromonas. Int. J. Syst. Bacte- Kamekura, M., Bardocz, S., Anderson, P., Wallace, R., and Kush- rioL, 45, 390-391. ner, D. J. (1986) Polyamines in moderately and extremely Esteve, C., Gutierrez, M.C., and Ventosa, A. (1995b) Aeromonas halophilic bacteria. Biochem. Biophys. Acta, 880, 204-208. enche/eia sp. nov., isolated from European eels. Int. J. Syst. Kita-Tsukamoto, K., Oyaizu, H., Nanba, K., and Simidu, U. (1993) Bacteriol., 45, 462-466. Phylogenetic relationships of marine bacteria, mainly members Gauthier, G., Gauthier, M., and Christen, R. (1995a) Phylogenetic of the family Vibrionaceae, determined on the basis of 165 analysis of the genera Alteromonas, Shewanella, and Moritella rRNA sequences. Int. J. Syst. Bacteriol., 43, 8-19. using genes coding for small-subunit rRNA sequences and divi- Lucchini, G.M. and Altwegg, M. (1992) rRNA gene restriction pat- sion of the genus Alteromonas into two genera, Alteromonas terns as taxonomic tools for the genus Aeromonas. Int. J. Syst. (Emended) and Pseudoalteromonas gen. nov., and proposal of Bacteriol., 42, 384-389. twelve new species combinations. Int. J. Syst. Bacteriol., 45, Martinez-Murcia, A. J., Benlloch, S., and Collins, M. D. (1992a) 755-761. Phylogeneric interrelationships of members of the genera Gauthier, M. J., Lafay, B., Christen, R., Fernandez, L., Acquaviva, Aeromonas and Plesiomonas as determined by 165 ribosomal M., Bonin, P., and Bertrand, J.-C. (1992) Marinobacter hydro- DNA sequencing: Lack of congruence with results of DNA- carbonoclasticus gen. nov., sp. nov., a new, extremely halotol- DNA hybridizations. Int. J. Syst. Bacteriol., 42, 412-421. erant, hydrocarbon-degrading marine bacterium. Int. J. Syst. Martinez-Murcia, A. J., Esteve, C., Garay, E., and Collins, M.D. Bacteriol., 42, 568-576. (1992b) Aeromonas allosaccharophila sp. nov., a new mes- Gauthier, G., Lafay, B., Ruimy, R., Breittmayer, V., Nicolas, J. L., ophilic member of the genus Aeromonas. FEMS Microbiol. Gauthier, M., and Christen, R. (1995b) Small-subunit rRNA se- Lett., 91,199-206. quences and whole DNA relatedness concur for the reassign- Mellado, E., Moore, E. R. B., Nieto, J. J., and Ventosa, A. (1995) ment of Pasteurel/a piscicida (Snieszko et al.) Janssen and Phylogenetic inferences and taxonomic consequences of 165 Surgalla to the genus Photobacterium as Photobacterium ribosomal DNA sequence comparison of Chromohalobacter damsela subsp. piscicida comb. nov. Int. J. Syst. Bacteriol., 45, marismortui, Volcaniella eurihalina and Deleya salina and re- 139-144. classification of V. eurihalina as Halomonas eurihalina comb. Hamana, K. (1996) Distribution of diaminopropane and acetylsper- nov. Int. J. Syst. BacterioL, 45, 712-716. midine in Enterobacteriaceae. Can. J. Microbiol., 42,107-114. Mellado, E., Moore, E. R. B., Nieto, J. J., and Ventosa, A. (1996) Hamana, K. and Matsuzaki, S. (1992a) Polyamines as a chemotax- Analysis of 165 rRNA gene sequences of Vibrio cost/cola onomic marker in bacterial systematics. Crit. Rev. Microbiol., strains: Description of Salinivibrio cost/cola gen, nov., comb. 18, 261-283. nov. Int. J. Syst. Bacteriol., 46, 817-821. Hamana, K. and Matsuzaki, S. (1992b) Taxonomic significance of Miller, J. M., Dobson, S. J., Franzmann, P. D., and McMeekin, T. A. polyamine synthesis in Paracoccus. J. Gen. Appl. Microbiol., (1994) Reevaluating the classification of Paracoccus ha/odeni- 38, 93-103. trificans with sequence comparisons of 165 ribosomal DNA. Hamana, K. and Matsuzaki, S. (1993) Polyamine distribution pat- Int. J. Syst. Bacteriol., 44, 360-361. terns serve as a phenotypic marker in the chemotaxonomy of Okamoto, T., Taguchi, H., Nakamura, K., Ikenaga, H., Kuraishi, H., the Proteobacteria. Can. J. Microbiol., 39, 304-310. and Yamasato, K. (1993) Zymobacter palmae gen. nov., sp. Hamana, K., Sakane, T., and Yokota, A. (1994) Polyamine analysis nov., a new ethanol-fermenting peritrichous bacterium isolated of the genera Aquaspirillum, Magnetospirillum, Oceanospirillum from palm sap. Arch. Microbiol., 160, 333-337. and Spirillum. J. Gen. AppL Microbiol. 40, 75-82. Okuzumi, M., Hiraishi, A., Kobayashi, T., and Fujii, T. (1994) Photo- Holmes, B. (1992) International committee on systematic bacteriol- bacterium histaminum sp. nov., a histamine-producing marine ogy subcommittee on the taxonomy of Vibrionaceae. Int. J. bacterium. Int. J. Syst. Bacteriol., 44, 631-636. Syst. Bacteriol., 42, 199-201. Ruimy, R., Breittmaayer, V., Eibaze, P., Lafay, B., Boussemart, M., Huys, G., Vancanneyt, M., Coopman, R., Janssen, P., Falsen, E., Gauthier, M., and Christen, R. (1994) Phylogenetic analysis Altwegg, M., and Kersters, K. (1994) Cellular fatty acid compo- and assessment of the genera Vibrio, Photobacterium, sition as a chemotaxonomic marker for the differentiation of Aeromonas, and Plesiomonas deduced from small-subunit phenospecies and hybridization groups in the genus rRNA sequences. Int. J. Syst. Bacteriol., 44, 416-426. Aeromonas. Int. J. Syst. Bacteriol., 44, 651-658. Smith, S. K., Sutton, D. C., Fuerst, J. A., and Reichelt, J. L. (1991) Ishimaru, K., Akagawa-Matsushita, M., and Muroga, K. (1995) Vib- Evaluation of the genus Listone//a and reassignment of Lis- rio penaeicida sp. nov., a pathogen of kuruma prawns. Int. J. tonella damsela (Love et al.) MacDonell and Colwell to the Syst. BacterioL, 45,134-138. genus Photobacterium as Photobacterium damsela comb. nov. Ishimaru, K., Akagawa-Matsushita, M., and Muroga, K. (1996) Vib- with an emended description. Int. J. Syst. Bacteriol., 41, 529- rio ichthyoenteri sp. nov., a pathogen of Japanese flounder 534. (Paralichthys olivaceus) Larvae. Int. J. Syst. BacterioL, 46, Sneath, P. H. A. (1993) Evidence from Aeromonas for genetic 155-159. crossing-over in ribosomal sequences. Int. J. Syst. BacterioL, Ivanova, E. P., Kiprianova, E. A., Mikhailov, V. V., Levanova, G. F., 43, 626-629. Garagulya, A. D., Gorshkova, N. M., Yumoto, N., and Yamamoto, S., Chowdhury, M. A. R., Kuroda, M., Nakano, T., 1997 Polyamines of Aeromonadaceae, Vibrionaceae, Pasteurellaceae, and Halomonadaceae 59

Koumoto, Y., and Shinoda, S. (1991) Further study on Norspermidine as a general constituent of Vibrio species. Can. polyamine compositions in Vibrionaceae. Can. J. MicrobioL, 37, J. Microbiol., 29, 724-728. 148-153. Yang, P., De Vos, P., Kersters, K., and Swings, J. (1993) Polyamine Yamamoto, S., Shinoda, S., Kawaguchi, M., Wakarnatsu, K., and patterns as chemotaxonomic markers for the genus Xan- Makita, M. (1983) Polyamine distribution in Vibrionaceae: thomonas. Int. J. Syst. Bacteriol., 43, 709-714.