INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, Oct. 1988, p. 424-429 Vol. 38, No. 4 0020-7713/88/040424-06$02.OO/O Copyright 0 1988, International Union of Microbiological Societies

Yersinia mollaretii sp. nov. and Yersinia bercovieri sp. nov., Formerly Called Yersinia enterocolitica Biogroups 3A and 3B GEORGES WAUTERS,l* MICHELE JANSSENS,' ARNOLD G. STEIGERWALT,2 AND DON J. BRENNER2 Microbiology Unit, Faculty of Medicine, Catholic University of Louvain, Brussels, Belgium, and Molecular Biology Laboratory, Meningitis and Special Pathogens Branch, Division of Bacterial Diseases, Centers for Disease Control, Atlanta, Georgia 303332

Yersiniu enterocolitica biogroups 3A and 3B are biochemically, serologically, and ecologically different from biogroup 3 and other Y. enterocolitica biogroups. Both biogroup 3A and biogroup 3B can be characterized by their negative Voges-Proskauer reactions and positive reactions in tests for pyrazinamidase, acid production from mucate, proline peptidase, and acid production from D-xylose. Biogroup 3A ferments L-sorbose but not L-fucose; biogroup 3B has the opposite fermentation pattern. Deoxyribonucleic acid relatedness studies (hydroxyapatite method) indicated that biogroups 3A and 3B are two new species that are about 55% interrelated and 25 to 46% related to other Yersiniu species (except Yersinia ruckeri [20 to 22%]). The names Yersiniu mollaretii sp. nov. and Yersinia bercovieri sp. nov. are proposed for biogroups 3A and 3B, respectively.

In addition to Yersinia enterocolitica sensu strict0 (3, Virulence markers. Autoagglutination after growth in several related species, previously known as Y. enteroco- broth at 37°C was tested in tryptic soy broth (17), and litica-like organisms, have been described. These are Yersi- calcium dependency was tested at 37°C on magnesium- nia intermedia (lo), Yersinia frederiksenii (19), Yersinia oxalate agar (14). kristensenii (8), Yersinia aldovae (7), and, most recently, Serotyping. 0 antigens were determined by the slide Yersinia rohdei (2). In 1978, Bercovier et al. described agglutination test, using 57 antisera against Y. enterocolitica strains originating from a terrestrial ecosystem and resem- and related Yersinia species (20). In addition, antisera were bling Y. enterocolitica biogroup 3 (4). The strains which raised against three new 0 factors designated 0:58, 059, fermented L-sorbose and either i- or myo-inositol were and 0:62. designated biogroup 3A; the strains which did not ferment DNA studies. Cells were grown at 37°C with shaking to these carbohydrates were designated biogroup 3B. In 1986, stationary phase in 1.5 liters of brain heart infusion broth Kaneko and Maruyama (16) isolated human pathogenic contained in 2-liter Erlenmeyer flasks. The methods used strains of serotype 3 in Japan that were Voges-Proskauer- for DNA isolation and purification, for DNA hybridization negative variants of biogroup 3. Typical serotype 3 strains by the hydroxyapatite method, and for guanine-plus-cy- are Voges-Proskauer positive and belong to biogroup 4. The tosine (G+C) content determinations have been described Voges-Proskauer-negative serogroup 3 strains were referred previously (11, 18). DNA was labeled in vitro with [32P] to as biogroup 3B by Fukushima (13). Wauters et al. (21) deoxyribose cytidine triphosphate by using a nick translation proposed that biogroup 3A and 3B strains be transferred to a reagent kit (Bethesda Research Laboratories, Inc., Gaithers- newly designated biogroup 6, to avoid confusion with bio- burg, Md.) and following the instructions of the manufac- group 3. turer. Differences in the ecology and the 0 and H antigenic patterns of biogroups 3A and 3B compared with biogroup 3 RESULTS prompted further biochemical and deoxyribonucleic acid DNA hybridization. Levels of DNA relatedness were (DNA) hybridization investigations of these strains. The determined by using labeled DNA from Y. mollaretii CNY results of these studies indicate that biogroups 3A and 3B are 7263T (T = type strain) and Y. bercovieri CNY 7506T (Table two new Yersinia species, for which the names Yersinia 2). All biogroup 3A strains tested were 74% or more related mollaretii (biogroup 3A) and Yersinia bercovieri (biogroup to strain CNY 7263T in both 60 and 75°C reactions (average, 3B) are proposed. 88% at 60"C, 89% at 75°C); the amount of divergence in related sequences was 0.5 to 2.5%. Similarly, with one MATERIALS AND METHODS exception, all biogroup 3B strains were at least 79% related to strain CNY 7506T in both 60 and 75°C reactions (average, Bacterial strains. The biogroup 3A and 3B strains which 86% at 60"C, 92% at 75°C); the amount of divergence in we used are listed in Table 1. The strains of other biogroups related sequences was 0.5%. The exception, biogroup 3B and other Yersinia species used in DNA hybridization stud- strain WE 120/83, was 56% related to Y. bercovieri at 60°C ies have been described previously (2,5, 7,8,19). All strains with 6.0% divergence and 41% related at 75°C; it was 39% were grown and maintained at room temperature on tryptic related to Y. mollaretii at 60°C with 9.0% divergence. The soy agar. levels of relatedness between Y. mollaretii and Y. bercovieri Biochemical properties. Biochemical tests were done by strains were 51 to 62% at 60°C with 5.0 to 7.5% divergence using the methods of Ewing (12) and Bercovier et al. (3). and 38 to 47% at 75°C. Y. mollaretii and Y. bercovieri were Pyrazinamidase activity (15) and proline peptidase (21) were 25 to 46% related to other Yersinia species (except Y. ruckeri tested as previously described. All tests were done at 25°C [20 to 22%]). Their levels of relatedness to Y. enterocolitica, and read at 1, 2, and 7 days, unless otherwise stated. including both Voges-Proskauer-positive and Voges-Pros- kauer-negative strains of biogroup 3, were no higher than * Corresponding author their levels of relatedness to most other Yersinia species.

424 VOL. 38, 1988 Y. MOLLARETII AND Y. BERCOVIERI 425

TABLE 1. Origins and serological characteristics of the Y. mollaretii and Y. bercovieri strains studied Strain" Source Country Received from: 0 antigensb Y. mollaretii WAIP 204T (= CDC Soil United States Bercovier 0:59,(20,36,7) 2465-87T = CNY 7263= = ATCC 43969T) WAIP 205 (= CNY 7264) Soil United States Bercovier 0:59,(20,36,7) WAIP 206 (= CNY 7442) Human stool The Netherlands Bercovier 0:59,(20,36,19) WA 291 Human stool Federal Republic of Germany Aleksic (H87-36/82) 0:3 WA 309 Raw vegetables France Catteau 0:59,(20,36,7) WA 739 Human stool The Netherlands Banffer 0:59,(20,36,19) WA749 Drinking water Federal Republic of Germany Ale ksic (H241-36/84) 0:6,30 WA 754 Human stool Finland Skurnik (10 Ye 92/84) 0: 59, (20,36,7) WA 751 Drinking water Federal Republic of Germany Aleksic (H253-36/84) 0:59,(20,36,19) WAT 932 Unknown Canada Toma (T9120) 0 :59, (20,36,7,19) WA942 Drinking water Federal Republic of Germany Aleksic (H412-36/83) 0:6,30 WA 989 Meat Austria Breuer 0:62,22 WE 316184 Human stool Belgium Nagels NT WS 19/86 Human stool Belgium Vandepitte 0:62,22 WS 20186 Human stool Belgium Van Noyen 0:7,13 WS 35/87 Human stool Belgium Van Noyen 0:62,22 W 787180 Human stool Belgium Wau ters 0:59,(20,36,7,19) Y. bercovieri WAIP 20gT (= CDC Human stool France Bercovier 0:58,16 2475-87T = CNY 7506' - ATCC 43970*) WAIP 207 (= CNY 7393) Human stool France Bercovier 0:58,16 WAIP 209 (= CNY 7966) Soil France Bercovier 0:18 WE 120183 Human stool Belgium Vandepitte 0:58,16 WA 313 Raw vegetables France Catteau 0:8 WA 314 Raw vegetables France Catteau 0:8 WA 315 Raw vegetables France Catteau 0:10 WA 317 Raw vegetables France Catteau Autoagglutinable WA 318 Raw vegetables France Catteau 0:5 WA 755 Human stool Finland Skurnik (12 Ye 127184) NT WA 760 Human stool Finland Skurnik (53 Ye 3016184) 0:58,16 WA 43/87 River water Norway Nesbakken 0:58,16 WE 187/80 Human stool Belgium Vandepitte 0:58,16 WE 580185 Human stool Belgium Wautele t 0:58,16 WE 171186 Human stool Belgium Collard 0:58,16 WE 83/86 Human stool Belgium Serruys 0:58,16 WE 188/86 Human stool Belgium Pepersack NT WE 68/87 Human stool Belgium Van Noyen 0:58,16 WA 763 Human stool Finland Skurnik 0:58,16 WS 235 Raw vegetables Belgium Wauters 0:58,16 W 601 Human stool Belgium Wauters 0:8 W 991 Human stool Belgium Wauters 0:58,16 WA 114 (= CNY 129) COYPU Great Britain Mair 0:8

a Abbreviations: W, WAIP, WA, WAT, WE, and WS, Laboratory in Brussels, Belgium; CDC, Centers for Disease Control, Atlanta; CNY, Centre National des Yersinia, Institut Pasteur, Paris, France; ATCC, American Type Culture Collection, Rockville, Md. The antigens in parentheses are weak or irregular. NT, Nontypable.

G+C contents. DNAs from Y. mollaretii CNY 7263T, Virulence markers. Tests for autoagglutination and cal- WAT 932, and WA 739 had G+C contents of 50,50, and 51 cium dependency at 37"C, indicating the presence of the mol%, respectively. DNAs from Y. bercovieri CNY 7506T, virulence plasmid in Y. enterocolitica, were negative for all WA 315, and WE 166/84 had G+C contents of 50 mol%. Y. mollaretii and Y. bercovieri strains tested. Assays were done in triplicate. Antigenic patterns. The 0 antigens found in both species Phenotypic characterization. Those characteristics com- are listed in Table 1. Some strains shared common 0 mon to all Y. mollaretii strains or all Y. bercovieri strains are antigens with Y. enterocolitica, especially 0:6,30 and 0:3 in given below in the species descriptions. Characteristics that Y. mollaretii and 0:s in Y. bercovieri. Cross-absorption vary within species are shown in Tables 3 and 4. Y. molla- studies showed that these common antigens were identical. retii ferments L-sorbose but not L-fucose. This distinguishes Most strains displayed one to three major new 0 factors it from Y. bercovieri, which has the opposite fermentation designated 058,059, and 0:62. Antigens 058 and 059 reactions. Characteristics that differentiated Y. mollaretii correspond to 0 factors provisionally designated C and B by and Y. bercovieri from other yersiniae are shown in Table 5, Aleksic et al. and described in Y.frederiksenii and biogroup and characteristics that differentiate the two new species 3A, respectively (1). Many Y. mollaretii strains possessed from Y. enterocolitica biogroups 3 and 4 are shown in Table antigen 059, and a few others had antigen 0:62. Most Y. 6. bercovieri strains had new factor 0:58. The other strains 426 WAUTERS ET AL. INT. J. SYST.BACTERIOL.

TABLE 2. Levels of DNA relatedness of Y. mollaretii and Y. bercovieri to Yersinia species Y. mollaretii CNY 7263T Y. bercovieri CNY 7506T Source of unlabeled DNA % Relatedness % % Relatedness % Relatedness % % Relatedness at 60°C Divergence" at 75°C at 60°C Divergence at 75°C Y. rnollaretii (biogroup 3A) CNY 7263T 100 0.0 100 62 7.5 38 WA 316184 94 0.5 97 CNY 7264 93 0.5 97 58 7.0 42 WA739 93 0.5 91 WA 751 92 1.0 92 CNY 7442 90 1.0 9i WAT 932 89 0.5 91 WA 989 79 2.5 76 WA 754 74 2.5 75 57 7.5 40 Y. bercovieri (biogroup 3B) CNY 7506T 56 7.0 44 100 0.0 100 WA 315 56 6.0 89 0.5 92 WA 755 51 6.0 88 0.5 94 WA 760 58 6.5 47 88 0.0 93 WE 166184 53 6.0 85 0.5 92 WE 187180 56 5.5 84 0.5 96 CNY 7966 55 5.0 79 0.5 87 WA 316 88 0.5 86 WE 580185 86 0.0 89 WE 120183 39 9.0 56 6.0 41 Y. enterocolitica 494-70 (biogroup 1) 37 44 211 (biogroup 2) 35 IP 6 (biogroup 3) 38 44 WA 29186 (biogroup 3) 36 40 IP 135 (biogroup 3) 35 40 39 (biogroup 3) 43 64 (biogroup 3) 37 175 (biogroup 4) 33 1144 (biogroup 5) 37 7706 (biogroup X1) 39 9.5 16 37 13.0 15 Y. aldovae 669-83 32 38 Y. frederiksenii 2581-77 45 46 6175 42 44 867 36 41 Y. intermedia 48 33 40 Y. kristensenii 1474 28 38 Y. rohdei 3022-85 22 40 Y. ruckeri 4535-69 22 20 Y. pseudotuberculosis P62 25 12 11 43 10.5 22 " Divergence (unpaired bases) in related sequences, calculated to the nearest 0.5%. TABLE 3. Variable characteristics of Y. mollaretii strains were either nontypable or were agglutinated to varying Cumulative % of strains Reaction of extents by various 0 antisera, but might have additional, Test positive on day:" type strain as-yet-unrecognized antigens. 0 1 2 7 CNY 7263& Several Y. mollaretii strains belonging to serogroup 0:s had a surface antigen called K2 (20). A few strains of both Motility at 36°C 6 6 6 species were tested for H antigens and were untypable by D-Glucose, gas 11 11 11 the available antisera. Acid produced from: D- Arabitol 0 6 6 Glycerol 65 94 100 DISCUSSION i- or myo-Inositol 0 6 23 Y. mollaretii and Y. bercovieri were first isolated from a Salicin 17 17 52 terrestrial ecosystem and were called Y. enterocolitica bio- Esculin hydrolysis 17 23 100 Acid produced from 41 88 94 groups 3A and 3B, respectively, by Bercovier et al. (4). Most mucate strains were isolated from environmental sources, but sev- Acetate utilization 52 52 100 eral others were of human origin, mainly from stools of both Nitratase A 6 healthy individuals and patients with diarrhea. There is no Nitratase B 94 evidence that these organisms are pathogenic for humans. Tetrat hionate 41 41 65 Their frequent occurrence in food, soil, water, and environ- a All tests were performed at 25°C. ment samples and the lack of virulence markers in these -, Negative after 7 days; +, positive within 2 days; (+), positive in 3 to species suggest a saprophytic character. 7 days. VOL. 38. 1988 Y. MOLLARETII AND Y. BERCOVIERI 427

TABLE 4. Variable characteristics of Y. bercovieri strains TABLE 6. Biochemical differentiation of Y. mollaretii and Y. bercovieri from Y. enterocolitica biogroups 3 and 4 Cumulative % of strains Reaction of Test positive on day:" type strain Y. enterocolitica CNY 7506b Y. rnol- Y. ber- 1 2 7 Test laretii covieri Biogroup Biogroup 3 Biogroup ~ ~~~~ 3 variant VP- 4 D-Glucose, gas 13 13 13 - Acid produced from: Voges-Proskauer - - + - + Glycerol 0 82 100 + Pyrazinamidase + + - - - Salicin 0 0 82 (+> Mutate, acid + + - - - Esculin hydrolysis 0 86 100 (+I Proline peptidase + + - a - - Acid produced from 47 95 100 + D-Xylose + + + + - mucate L-Sorbose + - + - + Acetate utilization 95 95 100 + L-Fucose - + - - -

a All tests were performed at 25°C. '' Some chinchilla strains are positive. -, Negative after 7 days; +, positive within 2 days; (+), positive in 3 to 7 days. enterocolitica (2), yersiniae appear to have species-specific H factors. Aleksic et al. (1) reported two new H factors, The new species conform to the descriptions of the family designated x and u, in Y. mollaretii (biogroup 3A). Specific H Enterobacteriaceae (9) and the genus Yersinia (6). Pheno- factors have not been reported for Y. bercovieri. typically, they most closely resemble each other and are DNA relatedness studies confirmed that biogroup 3A and next closest to Y. enterocolitica biogroups 3 and 4. As in biogroup 3B each represent a new species in the genus other Yersinia species, greater metabolic activity was evi- Yersinia, for which the names Y. mollaretii and Y. bercovieri dent at 25°C than at 36"C, especially in tests for L-ornithine are proposed. Biogroup 3B strain WE 120/83, although more decarboxylase, acetate utilization, and acid production from closely related to Y. bercovieri than to Y. mollaretii, was not mucate, in which reactions were sometimes delayed or related to Y. bercovieri at the species level (70% or more negative at 36°C. An occasional strain produced small relatedness at 60°C; 5% or less divergence in related se- amounts of gas in the tests for fermentation of glucose. For quences [22]). Since strain WE 120/83 cannot be phenotyp- more complete biochemical comparisons of Y. mollaretii and ically separated from Y. bercovieri, it is provisionally re- Y. bercovieri with other yersiniae the data presented here tained in this species. can be compared with previously published biochemical Description of Yersinia mollaretii sp. nov. Yersinia molla- descriptions (2, 5-8, 10, 19). It is important to distinguish Y. ret'ii (mol. la. ret' i. i. M. L. gen. n. mollaretii, in honor of mollaretii and Y. bercovieri from the mostly pathogenic Henri H. Mollaret, head of the National Yersinia Center at strains of Y. enterocolitica in biogroups 3 and 4 and from the the Pasteur Institute in Paris, France, whose many years of Voges-Proskauer-negative variants of biogroup 3 recently valuable studies have greatly increased our knowledge and described in Japan (16). The new species and Y. enteroco- appreciation of classification and epidemiology in the genus litica biogroups 3 and 4 are all negative in tests for indole, Yersinia [see references 2, 4-6, 8, 10, and 19 for references esculin, and Tween 80 esterase but are separable by the tests and citations to his work]) was formerly called Y. enteroco- shown in Table 6. litica biogroup 3A (4) and was later transferred to Y. entero- Some strains of Y. mollaretii and Y. bercovieri possess 0 colitica biogroup 6 (21). Cells are gram-negative, oxidase- antigens identical to those of Y. enterocolitica. Similar or negative, fermentative rods that are motile by peritrichous identical antigens have previously been documented in Y. flagella at 25°C but rarely motile at 36°C. The species enterocolitica, Y. intermedia, Y. frederiksenii, Y. kristen- conforms to the definitions of the 'family Enterobacteriaceae senii, Y. aldovae, and Y. rohdei (2, 5, 7, 8, 10, 19). Other (9) and the genus Yersinia (6).All strains grow on nutrient, than Y. rohdei, which contains the same H antigens as Y. tryptic soy, and MacConkey agar media. Whitish, opaque

TABLE 5. Biochemical differentiation of Y. mollaretii and Y. bercovieri from other Yersinia species Y. enterocolitica Y. mol- Y. ber- Y. inter- Y.fred- Y. kris- Y. aldo- y. rohdei pestis Y. pseudo- Test laretii covieri Biogroups 1 Biogroup media eriksenii tensenii vue tuberculosis y' ruckeri through 4 5 Indole -a - d + + Voges-Proskauer - - d + d Citrate (Simmons) - - - + d L-Ornithine + + + + + Mutate, acid + + - d d Pyrazinamidase + + d + + Sucrose + + + + + Cellobiose + + + + + L-Rhamnose - - - + + Melibiose - - - + - L-Sorbose + - d + + L-Fucose - + d d +

a +, Positive; -, negative; d, different reactions. ND, Not determined. Some reactions may be weakly positive. 428 WAUTERS ET AL. INT. J. SYST.BACTERIOL. colonies about 0.5 to 1.5 mm in diameter with smooth, entire deaminase, L-lysine decarboxylase, L-arginine dihydrolase, edges are formed on nutrient agar after 24 h at 25 or 36°C. gelatin hydrolysis, growth in KCN broth, malonate utiliza- Facultatively anaerobic. Good growth in 1day in tryptic soy, tion, Tween 8Q esterase, deoxyribonuclease, pigment pro- nutrient, and brain heart infusion broth media incubated at duction, and acid production from adonitol, D-arabitol, 25 or 36°C. More metabolically active at 25°C than at 36°C. dulcitol, erythritol, i- or myo-inositol, lactose, a-CH2-D- Positive reactions at 25°C in tests for methyl red, urea glucoside, raffinose, L-rhamnose, and L-sorbose. Acid pro- hydrolysis, L-ornithine decarboxylase, acid production from duction from salicin is delayed positive in most strains. A mucate, esculin hydrolysis (delayed in most strains), Jordan few strains produce small amounts of gas from D-glucose. tartrate, reduction of nitrates to nitrites (almost always The G+C content of the DNA is 50 mol%. contain nitratase B), o-nitrophenyl-P-D-galactopyranoside, Y. bercovieri strains have been isolated from human stool acetate utilization (often delayed), pyrazinamidase, proline specimens, animals, raw vegetables, soil, and water. There peptidase, and acid production from fermentation of D- is no evidence of pathogenicity for humans. The type strain glucose, L-arabinose, cellobiose, D-galactose, glycerol (may of Y. bercovieri is strain WAIP 208 (= CDC 2475-87 = CNY be delayed), maltose, D-mannitOl, D-mannose, D-sorbitol, 7506 = ATCC 43970). The G+C content of its DNA is 50 L-sorbose, sucrose, trehalose, and D-xylose. mol%. This strain was isolated from a human stool specimen Negative reactions in tests for indole production, the in France. The characteristics of the type strain are given in Voges-Proskauer reaction, Simmons citrate, H2S production Table 4. on Kligler iron agar and peptone iron agar, phenylalanine deaminase, L-lysine decarboxylase, L-arginine dihydrolase, LITERATURE CITED gelatin hydrolysis, growth in KCN broth, malonate utiliza- 1. Aleksic, S., J. Bockemuhl, and F. Lange. 1986. Studies on the tion, Tween 80 esterase, deoxyribonuclease, pigment pro- serology of flagellar antigens of Yersinia enterocolitica and duction, and acid production from adonitol, D-arabitol, related Yersinia species. Zentralbl. Bakteriol. Mikrobiol. Hyg. dulcitol, erythritol, L-fucose, lactose, melibiose, a-cH,-~- Abt. 1 Orig. Reihe A 261:299-310. glucoside, raffinose, and L-rhamnose. Tetrathionate reduc- 2. Aleksic, S., A. G. Steigerwalt, J. Bockemuhl, G. P. Huntley- tion is variable and is often delayed when positive, as are Carter, and D. J. Brenner. 1987. Yersinia rohdei sp. nov. isolated from human and dog feces and surface water. Int. J. acid production from i- or myo-inositol and salicin. A few Syst. Bacteriol. 37:327-332. strains produce small amounts of gas from D-glucose. The 3. Bercovier, H., J. M. Alonso, Z. N. Bentaiba, J. Brault, andH. H. G+C content of the DNA is 50 to 51 mol%. Mollaret. 1979. Contribution to the definition and the taxonomy Y. mollaretii strains have been isolated from human stool of Yersinia enterocolitica. Contrib. Microbiol. Immunol. 512- specimens, meat, raw vegetables, soil, and drinking water. 22. There is no evidence of pathogenicity for humans. 4. Bercovier, H., J. Brault, N. Barre, M. Treignier, J. M. Alonso, The type strain of Y. mollaretii is strain WAIP 204 (= CDC and H. H. Mollaret. 1978. Biochemical, serological and phage 2465-87 = CNY 7263 = ATCC 43969). The G+C content of typing characteristics of 459 Yersinia strains isolated from a its DNA is 50 mol%. It was isolated from soil in the United- terrestrial ecosystem. Curr. Microbiol. 1:353-357. 5. Bercovier, H., D. J. Brenner, J. Ursing, A. G. Steigerwalt, G. R. States. The characteristics of the type strain are given in Fanning, J. M. Alonso, G. P. Carter, and H. H. Mollaret. 1980. Table 3. Characterization of Yersinia enterocolitica sensu stricto. Curr. Description of Yersiniu bercovieri sp. nov. Yersinia berco- Microbiol. 4:201-206. vieri (ber. co. vi. e‘ ri. M. L. gen. n. bercovieri, in honor of 6. Bercovier, H., and H. H. Mollaret. 1984. Genus XIV. Yersinia HervC Bercovier, who first described biogroups 3A and 3B Van Loghem 1944, 15AL, p. 498-506. In N. R. Krieg and J. G. and has made outstanding contributions to the taxonomy and Holt (ed.), Bergey’s manual of systematic bacteriology, vol. 1. ecology of yersiniae [see references 3-8, 10, and 191) was The Williams & Wilkins Co., Baltimore. formerly called Y. enterocolitica biogroup 3B (4) and was 7. Bercovier, H., A. G. Steigerwalt, A. Guiyoule, G. Huntley- later transferred to biogroup 6 (21). Cells are gram-negative, Carter, and D. J. Brenner. 1984. Yersinia aldovae (formerly Yersinia enterocolitica-like group X2): a new species of Entero- oxidase-negative, fermentative rods that are motile by peri- bacteriaceae isolated from aquatic ecosystems. Int. J. Syst. trichous flagella at 25°C but not at 36°C. The species con- Bacteriol. 34: 166-172. forms to the definitions of the family’ Enterobacteriaceae (9) 8. Bercovier, H., J. Ursing, D. J. Brenner, A. G. Steigerwalt, G. R. and the genus Yersinia (6). Fanning, G. P. Carter, and H. H. Mollaret. 1980. Yersinia All strains grow on nutrient, tryptic soy, and MacConkey kristensenii: a new species of Enterobacteriaceae composed of agar media. Whitish, opaque colonies about 0.5 to 1.5 mm in sucrose-negative strains (formerly called atypical Yersinia en- diameter with smooth, entire edges are formed on nutrient terocolitica or Yersinia enterocolitica-like). Curr. Microbiol. 4: agar after 24 h at 25 or 36°C. Facultatively anaerobic. Good 2 19-224. growth in 1 day in tryptic soy, nutrient, and brain heart 9. Brenner, D. J. 1984. Family I. Enterobacteriaceae Rahn 1937, nom.fam.cons.opin. 15, Jud.Comm. 1958, 73; Ewing, Farmer, infusion broth media incubated at 25 or 36°C. More meta- and Brenner 1980, 674; Judicial Commission 1981, 104, p. 408- bolically active at 25°C than at 36°C. 420. In N. R. Krieg and J. G. Holt (ed.), Bergey’s manual of Positive reactions at 25°C in tests for methyl red, urea systematic bacteriology, vol. 1. The Williams & Wilkins Co., hydrolysis, L-ornithine decarboxylase, esculin hydrolysis Baltimore. (often delayed), acid production from mucate, tetrathionate 10. Brenner, D. J., H. Bercovier, J. Ursing, J. M. Alonso, A. G. reduction, Jordan tartrate, o-nitrophenyl-P-D-galactopy- Steigerwalt, G. R. Fanning, G. P. Carter, and H. H. Mollaret. ranoside, acetate utilization, nitrate reduction (nitratase B in 1980. Yersinia intermedia: a new species of Enterobacteriaceae all of the strains tested), pyrazinamidase, proline peptidase, composed of rhamnose-positive, raffinose-positive strains (for- and acid production from D-glucose, L-arabinose, cellobiose, merly called Yersinia enterocolitica or Yersinia enterocolitica- like). Curr. Microbiol. 4:207-212. L-fucose, D-galactose, glycerol (may be delayed), maltose, 11. Brenner, D. J., A. C. McWhorter, J. K. Leete Knutson, and D-mannose, D-sorbitol, sucrose, trehalose, and D-xylose. A. G. Steigerwalt. 1982. Escherichia vulneris: a new species of Negative reactions in tests for indole production, the Enterobacteriaceae associated with human wounds. J. Clin. Voges-Proskauer reaction, Simmons citrate, H2S production Microbiol. 15: 1133-1140. on Kligler iron agar and peptone iron agar, phenylalanine 12. Ewing, W. H. 1986. Edwards and Ewing’s identification of VOL. 38. 1988 Y. MOLLARETII AND Y. BERCOVIERI 429

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