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International Journal ofSystematic and Evolutionary Microbiology (2002),52,501-505 DOl: 10.1099/ijs.0.01836-0

Bacillus pycnus Spa nov. and neidei Spa NOTE nov., round-spored from soil

1 Microbial Properties L. K. Nakamura,1 O. Shida/ H. Takagi2 and K. Komagata3 Research Unit, National Center for Agricultural Utilization Research, 1815 N. University Street, Peoria, Author for correspondence: L K. Nakamura. Tel: + 13096816395. Fax: + 13096816672. IL 61604, USA e-mail: nakamulki"mail.ncaur.usda.gov

2 Research Laboratory, Higeta Shoyu Co. Ltd, Bacillus sphaericus sensu lato currently consists of seven or more groups of Choshi, Chiba 288, Japan unrelated taxa, one of which is B. sphaericus sensu stricto and another of 3 Department of which is Bacillus fusiformis. Members of two groups (groups 6 and 7), in Agricultural Chemistry, Tokyo University of common with all other B. sphaericus-like organisms, are unable to grow Agriculture, Setagaya-ku, anaerobically or to use common hexoses, pentoses and hexitols as sources of Tokyo 156, Japan carbon, have G+C contents of 34-36 mol % and form round spores. Groups 6 and 7 can be differentiated from other B. sphaericus-like organisms by low DNA relatedness and by variations in whole-cell fatty acid composition. Unique characteristics of group 6 include the ability to oxidize fi-hydroxybutyrate, the non-requirement for biotin and thiamin and failure to grow in 5 % NaCI. Distinctive traits of group 7 include the inability to oxidize pyruvate and a requirement for biotin, thiamin and cystine for growth. These data show that groups 6 and 7 represent two novel species, for which the names Bacillus pycnus sp. nov. and Bacillus neidei sp. nov., respectively, are proposed; the corresponding type strains are NRRL NRS-1691 T (= JCM 11075T) and NRRL BD-87T (= JCM 11077T).

Keywords: Bacillus PYC/lUS, Bacillus /leidei, novel species, 165 rDNA

The round-spored, mesophilic species Bacillus sphaeri­ comprised B. fils(jonnis: group I appeared to be a cus was first described by Neide (1904). Because early subgroup of 2. Groups 4-7 were unknown taxa. differentiation tests \vere ineffective, all round-spored In the present study, DNA relatedness evaluation and mesophiles were classified as B. sphaericus: the lack of phenotypic characterization were used to differentiate adequate differentiating tools deterred taxonomic groups 6 and 7 from each other, from representative study of mesophilic, round-spored bacteria. However, strains of groups 4 and 5 and from type strains of the discovery ofmosquitocidal activity among some of recognized round-spare-forming relatives. these organisms (Kellen et al., 1965) and the de­ velopment of molecular-biological techniques have DNA was extracted from 24-h cultures from groups I rekindled taxonomic and phylogenetic examination of (NRRL B-23269), 4 (NRRL NRS-593), 5 (NRRL T the round-spored organisms. Taxonomic heterogen­ B-1876), 6 (NRRL NRS-1691 , NRRL NRS-1693, eity of B. sphaericus had been suggested by variability NRRL NRS-1694, NRRL NRS-1695) and 7 (NRRL T of insecticidal activity among strains and was con­ BD-87 , NRRL BD-IOL NRRL BD-I03) and the firmed by comparative studies using DNA similarity following type cultures: 'Bacillus aminovorans' DSM T assessment (Krych et al., 1980), numerical 4337, B. fils(jormis DSM 2898 , globis­ T (Alexander & Priest, 1990: Priest et al., 1988), ran­ pora NRRL NRS-1533 , Bacillus insolitus NRRL T domly amplified polymorphic DNA fingerprinting NRS-1531 T, Marinibacillusmarinus DSM 1297 ,Sporo­ (Woodburn et al., 1995) and rRNA gene restriction sarcina psychrophila IFO 15381 T, B. silvestris DSM T T (Aquino de Muro et al., 1992). These developments led 12223 , B. sphaericus JCM 2502 and Ureibacillus T to the discovery of two more mesophilic species, thermosphaericus DSM 10633 . DNA reassociation Bacillus fils(lormis (Priest et al., 1988) and Bacillus was carried out using the method ofEzaki et al. (1989). silvestris (Rheims et al., 1999). In a phylogenetic study Probes were prepared with DNA from B. sphaericus T T based on 16S rDNA sequences (Nakamura, 2000), B. JCM 2502 , B. tllsijormis DSM 2898 , NRRL NRS­ T T sphaericus-like species segregated into seven groups 1691 • NRRL NRS-1694. NRRL BD-87 and NRRL wherein group 3 represented B. sphaericus and group 2 BD-I03.

01836 Printed in Great Britain 501 L. K. Nakamura and others

Group 1

!-=::::::::::==~-I Group 2 93 Bacillus fusiformis Group 3

99 B. sphaericus

Group 4 65 95 L-----I Group 5

'------B. silvestris NRS·1695 (AF169535) ] NRS·1691 T (AF169531) Group 6 ,..-__--"""00'-1 NRS·1694 (AF169534) (B. pycnussp. nov.) NRS·1693 (AF169533) 77 '-----i100 80-101 (AF169508) ] 82 Group 7 '-----1110<00 80-103 (AF169509) (B. neidei sp. nov.) 80-87 (AF169520) 89 L...... _ B. thermosphaericus P·11 (X90640)

...... ----1'00 B. psychrophilus ATCC 23304 T (X60634) B. globisporus OSM 4 T (X68415) ,..----1'00 L..... B. pasteurii NCIMB 8841 (X60631) L..... B. insolitus OSM 5 T (X60642) ...... ------'B. aminovorans'NCIMB 8292 (X62178) '------B. marinus OSM 1297 T (AJ237708) B. atrophaeus NCIMB 12899 (X60607) ,...------j1GG 99 B. subtilis NCOO 1769 (X60646) B. licheniformis OSM 13 T (X68416) '------B. cereus NCTC 11143 (X55063) ,...------B.lentus lAM 12466T (016272) UL------B. circulans lAM 12462 T (078312) '------B. megaterium lAM 13418T (016273) 76 ,...------B. stearothermophilus T10 (X57309) L- B. smithii OSM 4216T (X60643)

L.- ''"'oo''-j B. laevo/acticus lAM 12322 (016269) 7' 'B. myxolacticus'IAM 12326 (016274) ------r;;;;;--- Brevibacillus centrosporus NRRL NRS·664T (078458) ,... '00 T 72 B. brevis JCM 2503 (078457) T L------j'oo Aneurinibacillus migulanus ATCC 9999 (078462) A. aneurinilyticus NCIMB 10056 (X60645)

------G~======'00 Paenibacillus larvae NCDO 1141 (X60036) ~ P. polymyxa NCOO 1774 (60632) L..... Alicyclobacillus cycloheptanicus 1457 (X51928)

Fig_ 1. Neighbour-joining tree showing the phylogenetic position of groups 6 and 7 among the mesophilic and psychrophilic round-spored Bacillus species and selected oval-spored species. The tree is based on 1321-nt sequences. Confidence limits estimated from bootstrap analyses (500 replicates) appear at the nodes. This is a modification of the tree generated by Nakamura (2000). A maximum-parsimony tree generated from the sequence data showed a similar topology. Alicyclobacillus cycloheptanicus was designated the outgroup species for the analysis. B. thermosphaericus has been reclassified as Ureibacillus thermosphaericus (Fortina et al., 2001); B. globisporus, B. psychrophilus and B. pasteurii have been reclassified as Sporosarcina globispora, Sporosarcina psychrophila and , respectively (Yoon et al., 2001 a); B. marinus has been reclassified as Marinibacillus marinus (Yoon et al., 2001 b); and B. stearothermophilus has been reclassified as Geobacillus stearothermophilus (Nazina et al., 2001). Bar, 1 nt substitution per 10 nt.

Substrate oxidation profiles were obtained using the mercially prepared antibiotic discs on TGY agar plates BIOLOG GP system under conditions prescribed by spread-inoculated with the test organisms. The test the manufacturer. Other physiological traits were plates were incubated at 28 DC and the extent of examined using the methods of Gordon et al. (1973). inhibition was determined at 48 h. Decomposition Antibiotic sensitivity was assessed by placing com- of Tweens 40 and 80 was determined by the method

502 International Journal ofSystematic and Evolutionary Microbiology 52 Bacillus PYCllllS sp. nov. and B. neidei sp. nov. of Breuil & Gounot (1972). Vitamin and amino acid Table 1. Traits useful for differentiating the requirements were determined using the method of B. sphaericus-like groups Proom el al. (1955). ~ The BIOLOG system was used to evaluate substrate The extent of similarity among strains based on fatty oxidation. Substrates listed are those givin a 0 or 100 % acid composition data (Nakamura, 2000) was esti­ reactions. L-Glutamate was used bv all stn~ins: 64 substrates mated using the simple matching coefficient and were not used at all and variable r~actions were obtained with 26 substrates. AII organisms tested were sensitive to clustering was based on the unweighted pair group er~thromvcin. method with arithmetic averages (Sneath & SokaL chloramphenicol, tetracvcline and tobramvcin. Groups: 1 and 2 (n ~ 20), B..fils(fbrll1i~·: 3 (n = 7), . 1973). All cultures were grown on trypticase soy agar B..ljJ!IaericlIs: 4 (n = 10), unknown: 5 (n = 7). unknown: for 24 hat 30°C. Cell-wall peptidoglycan composition 6 (n = 3). B. pycnlls sp. nov.: and 7 (n = 4), B. neidei sp. nov. was analysed using the method described by Suzuki el al. (1993). Character Group In a phylogenetic tree based on 16S rDNA sequences, strains of groups 4-7 were positioned within a clade I and 2 3 4 5 6 7 that consisted generally of mesophilic, round-spored species, namely B. sphaericlls, B. jils(jorillis and Substrate oxidized: B. silveslris; the thermophile, U. lherillosphaericlls, is Pyruvate T + + + + situated outside the clade [Fig. 1; this tree is a fJ-Hydroxybutyrate + modification of one generated by Nakamura (2000)]. L-Alanine + + + + High bootstrap values validate the uniqueness of Glycyl L-glutamate + + + + + groups 4-7 within the clade of round-spored organ­ 2'-Deoxyadenosine + + + + + isms. Groups 6 and 7 were more closely related to Inosine + + + + each other than to the other mesophilic groups. AlvIP + + + UMP + + + High levels of DNA relatedness (97-100 %) were Growth in 5% NaCI + + T + + obser:ed between NRRL NRS-1691 T and other group Resistance to streptomycin + + 6 strams. namelv NRRL NRS-1693. NRRL NRS­ Decomposition of: 1694 and NRRL NRS-1695; 100% relatedness was TW'een 40 + + + + also observed between NRRL NRS-1694 and two Tween 80 + + + other group 6 strains (NRRL NRS-1693 and NRRL Vitamin requirement: NRS-1695). Low DNA relatedness values. of 27 and Biotin 19 %, respectively, were determined between NRRL + + + + + Thiamin NRS-1694 and two group 7 strains, NRRL BD-87T + + + + + Cystine requirement and NRRL BD-I03. Similarlv. low levels of DNA + relatedness were observed bet\~een NRRL NRS-1694 and representative strains from groups 1 (NRRL B­ 23269),4 (NRRL NRS-593) and 5 (NRRL B-1876): data show that groups 6 and 7 are genetically distinct values were respectively 23, 27 and 15 %. Finally, the taxa. levels of DNA relatedness between NRRL NRS-1694 and the type strains of existing round-spored species Phenotypic data collated from this and previous CB. aillinovorans' DSM 4337, B. fils(forillis DSM studies (Table 1) demonstrated the phenotypic distinc­ T T 2898 • S. globispora NRRL NRS-1533 , B. insolilus tiveness of groups 6 and 7. Traits that differentiate T T NRRL NRS-1531 , M. lIlarinus DSM 1297 , Sporo­ group 6 from all the other taxa were j1-hydroxybutyrate T sarcina pasleurii DSM 33 , S. psychrophila IFO oxidation, growth inhibition by 5 % NaCI and non­ T 15381 T, B. silveslris DSM 12223 , B. sphaericus JCM requirement for vitamins. Unique traits of group 7 T T 2502 and U. lherillosphaericlis DSM 10633 ) ranged were failure to oxidize pyruvate and a growth re­ from 1 to 18 %. quirement for cystine. The dendrogram based on fatty acid composition DNA relatedness values of 100 % were measured shown in Fig. 2 shows clear separation ofgroups 6 and between NRRL BD-87T and other group 7 strains 7 from each other, from groups 4 and 5 and from the (NRRL BD-I01 and NRRL BD-103). The levels of existing named mesophilic species. Numerical analv­ DNA relatedness between NRRL BD-I0l and NRRL sis of substrate-oxidation patterns segregated tl;e BD-I03 were also 100 %. Low DNA relatedness values organisms similarly (Nakamura, 2000). The predomi­ were noted between NRRL BD-87T and representative nant fatty acids found in group 6 were 15: Oiso strains from groups I (NRRL B-23269), 4 (NRRL (70'3%), 15:0anteiso (8'1 %) and 16: IOJ7cis alcohol NRS-593) and 5 (NRRL B-1876); the respective values (6'0%). Smaller amounts of 16:0 (3'1 %) and 16: were 20, 17 and 19 %. Low levels of DNA relatedness. IOJ 11 cis (1-4 %) fatty acids were also found. ranging from 3 to 19 %, were measured between NRRL BD-87T and the type strains of the existing In group 7, the most abundant fatty acids were 15: Oiso round-spored species listed above. DNA relatedness (23'6%), 15:0anteiso (17'8%) and 16:1OJllcis http://ijs.sgmjournals.org 503 L. K. Nakamura and others

Group Acetylmethylcarbinol, dihydroxyacetone. indole and 80·94 NR5-967 HzS are not produced. The pH in Voges-Proskauer 80-83 B-183 ~~ ~ broth ranges from 7·2 to 7·6. Starch. casein. tyrosine. 80-95 ] .. 8-23268 .. urea. Tweens 40 and 80 and egg-yolk lecithin are not 80-107 84297 NRS-719 decomposed. Common hexoses. pentoses. hexitols. NRS-810 NR$.111 4 disaccharides and trisaccharides are not fermented. 80·115 80·117 Grows at pH 5'7. but not in the presence of 0·001 % NRS·1692 ] 80·119 lysozyme and 5 % NaCI. Sensitive to chloramphenicol, B-23279 B-23283 8-23284 tobramycin. streptomycin. erythromycin and tetra­ 8-23285 8-23286 cycline. Biotin. thiamin and cystine are not required 8-23287 8-23269 for growth. Based on the BIOLOG GP method. 8·14957 8-23280 pyru~ate and fJ-hydroxybutyrate are oxidized. Citrate. 8-23281 propionate, L-alanine. glycyl L-glutamate, 2'-deoxy­ ~fff(fI3 adenosine. inosine. AMP and UMP are not oxidized. ] (B.fus7formiS) Optimum growth temperature is 28-30 °C (minimum. NRS-1756 NRS·71B 8·14905 5-10 °C: maximum. 40-45 0C). Whole-cell fatty acids NRS·1691 ] ~~tm~ detected are 15: Oiso (70'3 %), 15: Oanteiso (8'1 %). 16: 6 1w7cisalcoho1(6·0%).16:0iso(3·1 %)and 16: Iwllcis r----!~J:~~:: (1-4%). Cell wall peptidoglycan type is L-Lys-D-Glu. ] 5 '----NRS·llB9 Isolated from soil, G+C content is 35 mol%. Type BO-93 T T BO-B7 ] strain is NRRL NRS-1691 (= JCM 11075 ). ::~~~~~:=~~~5~~B-1876BO·101 7 ~ 80·103 I II I II I II I I I gj 6.7 4£ 2.4 02 Description of Bacillus neidei sp. nov. Bacillus neidei (nei/de.i. N.L. gen. n. neidei ofNeide, in recognition of the early microbiologist E. Neide). Fig. 2. Dendrogram showing the relationship of groups 6 and 7 to the other B. sphaericus-like species. A matrix consisting of Rods are about 1·0 x 3,0-5,0 ~lm (determined by photo­ 47 strains and 12 fatty acids was used. The fatty acid com­ micrograph). Gram-positive. Motile. Round spores' positional data were obtained in a previous study (Nakamura, form in swollen sporangia. Agar colonies are non­ 2000). Similarity was estimated by means of the simple matching coefficient (Ssm) and clustering was based on the unweighted pigmented, translucent. thin. smooth. circular. entire pair group method with arithmetic averages (Sneath & Sokal, and the mean diameter is about 1 mm after 24 h of 1973). incubation on TGY at 28°C. Catalase is produced. Strictly aerobic. Nitrate is not reduced to nitrite. Acetylmethylcarbinol, dihydroxyacetone. indole and (14'5 %). Other fa tty acids observed were 14: Oiso HzS are not produced. The pH in Voges-Proskauer (5,0%). 15:0 (1'7%). 16: 1w7cis alcohol (4'7%) and broth ranges from 7·2 to 7·6. Starch. casein. tyrosine. 16:0iso (8'1 %). urea. Tweens 40 and 80 and egg-yolk lecithin are not decomposed. Common hexoses. pentoses. hexito1s. L-Lysine and D-glutamic acid were the key amino acids disaccharides and trisaccharides are not fermented. found in the cell wall peptidoglycans of NRRL NRS­ T T Grows at pH 5·7 and in 5 % NaCL but not in the 1691 and NRRL BD-87 . Like many of the round­ presence of 0·001 % lysozyme. Sensitive to chloram­ spored species (Stackebrandt et al.• 1987). the pep­ phenicoL tobramycin. streptomycin. erythromycin tidoglycan type for groups 6 and 7 was L-Lys-D-Glu. and tetracycline. Biotin, thiamin and cystine are Based on these results. groups 6 and 7 merit recog­ required for growth. Based on the BIOLOG GP nition as two novel species. for which the l:espectiv~ method. pyruvate and fJ-hydroxybutyrate are oxidized. / names Bacillus PYCl1US sp. nov. and Bacillw;.:neidei sp.. Citrate. propionate. L-a1anine. glycyl L-glutamate. 2 ­ nov. are proposed. The respective type strains are deoxyadenosine. inosine. AMP and UMP are not r T NRRL NRS-169r (= JCM I 1075 ) and NRRL BD­ oxidized. Optimum growth temperature is 28-30°C T T 87 (= JCM IIOn ). (minimum. 5-10 °C: maximum. 40-45 0C). Whole-cell fatty acids detected are 14:0iso (5'0%).15:0 (1'7%). Description of Bacillus pycnus sp. nov. 15:0iso (23'6%). 15:0anteiso (17'8%),16:0 (5-4%), 16: 1w7cis alcohol (4'7%). 16:0iso (8'1 %) and 16: Bacillus PYCl1US (pyc/nus. Gr. adj. pykl10S thick: N.L. 1w11cis (14'5 %). Cell wall peptidoglycan type is adj. PYCl1US thick. referring to thick cells). L-Lys-D-Glu. Isolated from soil, G+C content is Rods are 1,0-1,5 x 3,0-5,0 ~lm (determined from pho­ 35 mol %. Type strain is NRRL BD-87T (= JCM T tomicrograph). Gram-positive. Motile. Round spores 110n ). form in swollen sporangia. Agar colonies are non­ pigmented. translucent. thin. smooth. circular. entire References and the mean diameter is about I mm after 24 h of Alexander, B. & Priest, F. G. (1990). Numerical classification and incubation on TGY agar at 28°C. Catalase is pro­ identification of Bacill/ls .Ip!lacric/ls including some strains pathogenic duced. Strictly aerobic. Nitrate is not reduced to nitrite. for mosquito larvae. J Gcn Microbiol 136. 367-376.

504 International Journal of Systematic and Evolutionary Microbiology 52 Bacillus PYCIIUS sp. nov. and B. neidei sp. nov.

Aquino de Muro, M., Mitchell, W. J. & Priest, F. G. (1992). Neide, E. (1904). Botanische Beschreibung einiger sporenbildenden Differentiation of mosquito-pathogenic strains of Bacillus .Iphaericus Bakterien. Zelllbl Bakreriol Parasirenkd Infekrionskr Hyg Abr /I 12. from non-toxic varieties by ribosomal RNA gene restriction patterns. 337-352. j Geu Mierobiol 138. 1159-1166. Priest, F. G., Goodfellow, M. & Todd, C. (1988). A numerical Breuil, C. & Gounot, A. M. (1972). Recherches preliminaries sur les classification of the genus Bacil/us. j Gen Microbiol 134. 1847-1882. bacteries lipolytiques psychrophiles des sols et des eaux. Can j Microbiol Proom, H. & Knight, B. C. J. G. (1955). The minimal nutritional 18. 1445-145l. requirements of some species in the genus Bacillus. j Gen Jlierobiol 13. Ezaki, T., Hashimoto, Y. & Yabuuchi, E. (1989). Fluorometric 474--480. deoxyribonucleic acid-deoxyribonucleic acid hybridization in micro­ Rheims, H., Friihling, A., Schumann, P., Rohde, M. & Stackebrandt, dilution wells as an alternative to membrane filter hybridization in E. (1999). Bacil/us si/resrris sp. nov.. a new member ofthe genus Bacif/us which radioisotopes are used to determine genetic relatedness among that contains lysine in its cell wall. IIIl j Sysr Bacieriol49. 795-802. bacterial strains. IIIl j Sysr Baueriol39. 224-229. Sneath, P. H. A. & Sokal, R. R. (1973). Numerical Tcc-.:onomy. San Fortina, M. G., Pukall, R., Schumann, P., Mora, D., Parini, c., Francisco: W. H. Freeman. Manachini, P. L. & Stackebrandt, E. (2001). Ureibacif/us gen. nov.. a Stackebrandt, E., Ludwig, W., Weizenegger, M., Dorn, S., McGill, new genus to accommodate Bacil/us rhermo.lphaericus (Andersson er 01. T. 1, Fox, G. E., Woese, C. R., Schubert, W. & Schleifer, K.-H. 1995). emendation of (.ireibacillus rhermo.lplwericus and description of (1987). Comparative 16S rRNA oligonucleotide analyses and murein Ureibacillus rerrenus sp. nov. IIIl j Sl'sr £1'01 Mierobiol 51. 447--455. types of round-spore-forming and non-spore-forming relatives. Gordon, R. E., Haynes, W. C. & Pang, C. H.-N. (1973). Genus Bacillus. j Gen Mierobiol 133. 2523-2529. Agricu/rure Handbook no. 427. Washington. DC: United States De­ Suzuki, K., Goodfellow, M. & O'Donnell, A. G. (1993). Cell partment of Agriculture. envelopes and classification. In Handbook ofNe\1' Bauerial Sysremarics. Kellen, W. R., Clark, T. B., Lindegren, J. E., Ho, B. c., Rogoff, M. H. pp. 195-250. Edited by Ivi. Goodfellow & A. G. O·Donneli. London: Academic Press. & Singer, S. (1965). Bacil/us sphaericus Neide as a pathogen of mosquitoes. j Imerrebr Parlrol 7.442--448. Woodburn, M. A., Yousten, A. A. & Hilu, K. H. (1995). Random amplified polymorphic DNA fingerprinting of mosquito-pathogenic Krych, V. K., Johnson, J. L. & Yousten, A. A. (1980). Deoxy­ and nonpathogenic strains of Bacil/us .Iphaericus. IIIl j Sysr Bacreriol45. ribonucleic acid homologies among strains of Bacillus sphaericus. IIIl j 212-217. Sysr Bacreriol 30. 476--484. Yoon, J.-H., Lee, K.-C., Weiss, N., Kho, Y. H., Kang, K. H. & Park, Nakamura, L. K. (2000). Phylogeny of Bacif/us sphaericus-like organ­ Y.-H. (2001a). Sporosarcina a(juimarina sp. nov.. a bacterium isolated isms.11Il j Sysr £1'01 Mierobiol 50.1715-1722. from seawater in Korea. and transfer of Bacillus globisporus (Larkin Nazina, T. N., Tourova, T. P., Poltaraus, A. B. & 8 other authors and Stokes 1967). Bacillus psyc/Irophilus (Nakamura 1984) and Bacif/us (2001). Taxonomic study of aerobic thermophilic bacilli: descriptions pasreurii (Chester 1898) to the genus Sporosarcina as Sporosarcina of Geobacillus subrerraneus gen. nov.. sp. nov. and Geobacif/us u:encnsis globi.\pora comb. nov.. Sporosarcina psychrop/Iila comb. nov. and sp. nov. from petroleum reservoirs and transfer of Bacif/us srearo­ Sporosarcina pasreurii comb. nov.. and emended description of the rhermophilus. Bacillus rhernrocar(,/l/Ilarus. Bacillus rhermoleororans. genus Sporosarcina. IIIl j Sysr £1'01 Mierobiol 51. 1079-1086. Bacillus kausrophilus. Bacif/us rhermoglucosidasius and Bacillus rhermo­ Yoon, 1-H., Weiss, N., Lee, K.-C., Lee, 1.-5., Kang, K. H. & Park, denirrincans to Geobacillus as the new combinations G. srearo­ Y.-H. (2001b). jcorga!ibacif/us a!imclllarius gen. nov.. sp. nov.. a novel rhet/nophilus. G. rhermocarcmr!aills. G. rhermoleororans. G. kausro­ bacterium isolated from jeotgal with L-Iysine in the cell wall. and philus. G. rhermoglucosidasius and G. rhermodenirrificans.11Il j Sysr £1'01 reclassification ofBacillus marinus Ruger 1983 as Marinibacillus maril/us Mierobiol 51. 433--446. gen. nov.. comb. nov. IIIl j Sysr £1'01 Mierobiol 51. 2087-2093.

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