lnternationsl Journal of Systematic Bacteriology zyxwvuts (1 998), 48, 5 1 1-5 1 8 zyxwvut Printed in Great Britain Staphylococcus succinus zyx sp. nov., isolated f rorn Dominican amber zyx L. H. Lambert,’ T. Cox,’ K. Mitchell,’ R. A. Rossell6-M0ra,~ C. Del C ~ e t o , ~ D. E. Dodge,5 P. Orkand4 and R. J. Cano2 Author for correspondence: L. H. Lambert. Tel: + 1 510 644 1170 ext. 2014. Fax: + 1 510 841 7805. 1 XOMA Corporation, 2910 Seventh Street, Berkeley, CA94710, USA 2 Environmental Biotech nology Institute, Ca I iforn ia Polytechnic State University, San Luis Obispo, CA 93407, USA zyxwvutsrqp 3 lnstituto Mediterraneo de Estudios Avanzados (CSIC- UIB), Crtra. Valldemosa Km 7.5, Palma de Mallorca, Spain 4 lnstituto de Neurobiologia, Universidad de Puerto Rico, San Juan, PR 00901, Puerto Rico 5 PE Applied Biosystems, 850 Lincoln Centre Dr., Foster City, zyxwvutsrqp CA 94404, USA Two bacterial isolates, designated AMG-D1 and AMG-D2, were recovered from 25-35-million-year-old Dominican amber. AMG-D1 and AMG-D2 biochemically most closely resemble Staphy/ococcus Xylosus; they differ physiologically from other staphylococci. Fatty acid analysis and comparisons with extensive databases were unable to show relatedness to any specific taxon. Moreover, AMG-D1 and AMG-D2 contain tuberculostearic acid and meso-diaminopimelic acid, characteristic of the G+C-rich coryneform bacteria, as opposed to L-lysine characteristic of staphylococci. AMG-DIT and AMG-D2 have a G+C ratio of 35 mol%. Phylogenetic analysis with the 16s rRNA gene indicated that AMG- DIT and AMG-D2 were most closely related to Staphy/ococcus equomm, 5. Xylosus, Staphy/ococcus saprophyticus and other n ovo b i oc i n-res i sta n t staphylococci. Stringent DNA-DNA hybridization studies with AMG-D1 revealed similarities of 38% with S. equonrrn, 23% with S. xylosus and 6% with 5. saprophyticus. The results indicate that AMG-DIT and AMG-D2 represent a novel species, which was named Staphy/ococcus succinus sp. nov. The type strain of the new species is AMG-D1 (ATCC 700337). Keywords : zyxwv Staphylococcus succinus sp. nov., Dominican amber INTRODUCTION Amber, a polymeric glass formed over time from the resins of conifers and some flowering plants, provides an excellent preservative matrix for biological speci- mens. Can0 & Borucki reported the recovery of a viable Bacillus sphaericus-like isolate from the gut of a stingless bee found in Dominican amber estimated to be 25-40 million years old (3, 17). Observations made during the recovery of this spore-former were suf- ficiently compelling to attempt the recovery of non- spore-forming bacteria from inclusions in amber obtained from the same Dominican source. Members of the genus Staphylococcus are ubiquitous and diverse. Currently, 29 recognized species isolated from soil, plants, animals and humans have been described (4, 14, 24, 26, 31). Subjected to extreme environmental conditions, such staphylococci are Abbreviations: DAP, rneso-diaminopimelic acid; FAME, fatty acid methyl ester. The GenBank accession numbers for the sequences reported in this paper are AF004219 (AMG-D13 and AF004220 (AMG-D2). unusually durable. Viable organisms were recovered from cultures exposed to low temperatures (1 10-3 10 K), salt and other desiccating agents (32,37). After exposure to a barrage of protons corresponding to about 250 years in solar space, Staphylococcus aureus showed very high survival rates (1 5). Addition- ally, Potts (25) has reported the recovery of viable Gram-positive, non-spore-forming bacilli and cocci from Pliocene permafrost sands and other air-dried environments. Here we describe two Staphylococcus-like strains, AMG-DIT and AMG-D2, which were isolated from plant and soil inclusions in Dominican amber. Using a polyphasic approach, including biochemical, physio- logical, morphological and molecular data zy (6), we grouped these two strains into a new species, Staphy- lococcus succinus. METHODS Amber decontamination. A piece of Dominican amber, measuring approximately 3 x 7 x 2 cm and containing small inclusions with the appearance of plant and soil debris, was examined for the presence of cracks or fissures, then rinsed 00659 zyxwvutsrqponm 0 1998 IUMS 51 1