We describe two novel species of Acholeplasma sp. strain N93 and Mycoplasma sp. strain LR5794 which were isolated from the nasopharynx of a horse from the United Kingdom and from the oral cavity of a North American raccoon from Canada, respectively. These strains were phenotypically and genetically characterized and compared to other established Mycoplasma and Acholeplasma species. Both strains are facultative anaerobes, resistant to penicillin, and produce acid from glucose but do not hydrolyze arginine and urea. Both strains grew well in microaerophilic and anaerobic atmospheric conditions at 35–37 °C using PPLO (pleuropneumonia-like organisms) medium. Acholeplasma sp. N93 does not require serum for growth. Colonies of both strains showed a typical fried-egg appearance and transmission electron microscopy of bacterial cells revealed a typical mycoplasma cellular morphology. Molecular characterization included assessment of several genetic loci. The genetic analysis indicated that Acholeplasma sp. N93 and Mycoplasma sp. LR5794 were most closely related to A. hippikon and A. equifetale, and M. molare and M. lagogenitalium, respectively. However, both novel strains were genetically unique in comparison to other well-known Mycoplasma and Acholeplasma species. Based on the isolation source history, phenotypic, genotypic, and phylogenetic characteristics of these novel strains, we propose the name Acholeplasma equirhinis sp. nov. for Acholeplasma sp. isolated from the nasopharynx of a horse [the type strain is N93T (= DSM 106692T = ATCC TSD-139T = NCTC 14351T)], and the name Mycoplasma procyoni sp. nov. for the Mycoplasma sp. isolated from the oral cavity of a North American raccoon [the type strain is LR5794T (= DSM 106703T = ATCC TSD-141T = NCTC 14309T)].
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Adekambi T, Shinnick TM, Raoult D, Drancourt M (2008) Complete rpoB gene sequencing as a suitable supplement to DNA-DNA hybridization for bacterial species and genus delineation. Int J Syst Evol Microbiol 58:1807–1814
Allam NM, Lemcke RM (1975) Mycoplasmas isolated from the respiratory tract of horses. J Hyg (Lond) 74:385–408
Angulo AF, Reijgers R, Brugman J, Kroesen I, Hekkens FE, Carle P, Bove JM, Tully JG, Hill AC, Schouls LM, Schot CS, Roholl PJ, Polak-Vogelzang AA (2000) Acholeplasma vituli sp. nov., from bovine serum and cell cultures. Int J Syst Evol Microbiol 50:1125–1131
Bronstad A, Berg AG (2011) The role of organizational culture in compliance with the principles of the 3Rs. Lab Anim 40:22–26
Brown DR (2010) Phylum XVI. Tenericutes Murray 1984a, 356VP (Effective publication: Murray 1984b, 33). In: Bergey's manual of systematic bacteriology, vol 4, 2nd edn. Springer, New York, pp 567–723
Brown DR, Whitcomb RF, Bradbury JM (2007) Revised minimal standards for description of new species of the class Mollicutes (division Tenericutes). Int J Syst Evol Microbiol 57:2703–2719
CABI (2019) Procyon lotor (raccoon): datasheet. https://www.cabi.org/isc/datasheet/67856
Chun J, Oren A, Ventosa A, Christensen H, Arahal DR, da Costa MS, Rooney AP, Yi H, Xu XW, De Meyer S, Trujillo ME (2018) Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol 68:461–466
Firrao G, Brown DR (2013) International Committee on Systematics of Prokaryotes. Subcommittee on the taxonomy of Mollicutes: Minutes of the meetings, July 15th and 19th 2012, Toulouse, France. Int J Syst Evol Microbiol 63:2361–2364
Fournier PE, Suhre K, Fournous G, Raoult D (2006) Estimation of prokaryote genomic DNA G+C content by sequencing universally conserved genes. Int J Syst Evol Microbiol 56:1025–1029
Gasparich GE, Kuo CH (2019) Genome analysis-based union of the genus Mesoplasma with the genus Entomoplasma. Int J Syst Evol Microbiol 69:2735–2738
Glass JI, Assad-Garcia N, Alperovich N, Yooseph S, Lewis MR, Maruf M, Hutchison CA, Smith HO, Venter JC (2006) Essential genes of a minimal bacterium. Proc Natl Acad Sci USA 103:425–430
Harasawa R, Imada Y, Ito M, Koshimizu K, Cassell GH, Barile MF (1990) Ureaplasma felinum sp. nov. and Ureaplasma cati sp. nov. isolated from the oral cavities of cats. Int J Syst Bacteriol 40:45–51
Kampfer P, Glaeser SP (2012) Prokaryotic taxonomy in the sequencing era—the polyphasic approach revisited. Environ Microbiol 14:291–317
Kirchoff H (1974) Neue Spezies der Fam. Acholeplasmataceae und der Fam. Mykoplasmataceae bei Pferden. Zentralbl Veterinarmed [B] 21:207–210
Konstantinidis KT, Tiedje JM (2005) Towards a genome-based taxonomy for prokaryotes. J Bacteriol 187:6258–6264
Konstantinidis KT, Ramette A, Tiedje JM (2006) The bacterial species definition in the genomic era. Philos Trans R Soc Lond B Biol Sci 361:1929–1940
Kotani H, Ogata M, Koshimizu K, Tashiro K, Masui M (1982) Isolation of Acholeplasma laidlawii from a giraffe (Giraffa camelopardalis rothschildi) and a mouflon (Ovis musimon). Vet Rec 110:16
Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874
May M, Brown DR (2019) International Committee on Systematics of Prokaryotes Subcommittee on the taxonomy of Mollicutes: minutes of the closed meeting, 8 July 2018, Portsmouth, New Hampshire, USA. Int J Syst Evol Microbiol 69:2169–2171
Mollet C, Drancourt M, Raoult D (1997) rpoB sequence analysis as a novel basis for bacterial identification. Mol Microbiol 26:1005–1011
Ochman H, Elwyn S, Moran NA (1999) Calibrating bacterial evolution. Proc Natl Acad Sci USA 96:12638–12643
Poveda JB (1998) Biochemical characteristics in mycoplasma identification. Methods Mol Biol 104:69–78
Ramirez AS, Vega-Orellana OM, Viver T, Poveda JB, Rosales RS, Poveda CG, Spergser J, Szostak MP, Caballero MJ, Ressel L, Bradbury JM, Mar Tavio M, Karthikeyan S, Amann R, Konstantinidis KT, Rossello-Mora R (2019) First description of two moderately halophilic and psychrotolerant Mycoplasma species isolated from cephalopods and proposal of Mycoplasma marinum sp. nov. and Mycoplasma todarodis sp. nov. Syst Appl Microbiol 42:457–467
Tully JG, Whitcomb RF, Hackett KJ, Rose DL, Henegar RB, Bove JM, Carle P, Williamson DL, Clark TB (1994a) Taxonomic descriptions of eight new non-sterol-requiring mollicutes assigned to the genus Mesoplasma. Int J Syst Bacteriol 44:685–693
Tully JG, Whitcomb RF, Rose DL, Bove JM, Carle P, Somerson NL, Williamson DL, Eden-Green S (1994b) Acholeplasma brassicae sp. nov. and Acholeplasma palmae sp. nov., two non-sterol-requiring mollicutes from plant surfaces. Int J Syst Bacteriol 44:680–684
Volokhov DV, George J, Liu SX, Ikonomi P, Anderson C, Chizhikov V (2006) Sequencing of the intergenic 16S–23S rRNA spacer (ITS) region of Mollicutes species and their identification using microarray-based assay and DNA sequencing. Appl Microbiol Biotechnol 71:680–698
Volokhov DV, Neverov AA, George J, Kong H, Liu SX, Anderson C, Davidson MK, Chizhikov V (2007) Genetic analysis of housekeeping genes of members of the genus Acholeplasma: phylogeny and complementary molecular markers to the 16S rRNA gene. Mol Phylogenet Evol 44:699–710
Volokhov DV, Simonyan V, Davidson MK, Chizhikov VE (2012) RNA polymerase beta subunit (rpoB) gene and the 16S–23S rRNA intergenic transcribed spacer region (ITS) as complementary molecular markers in addition to the 16S rRNA gene for phylogenetic analysis and identification of the species of the family Mycoplasmataceae. Mol Phylogenet Evol 62:515–528
Volokhov DV, Batac F, Gao Y, Miller M, Chizhikov VE (2019a) Mycoplasma enhydrae sp. nov. isolated from southern sea otters (Enhydra lutris nereis). Int J Syst Evol Microbiol 69:363–370
Volokhov DV, Gulland FM, Gao Y, Chizhikov VE (2019b) Ureaplasma miroungigenitalium sp. nov. isolated from northern elephant seals (Mirounga angustirostris) and Ureaplasma zalophigenitalium sp. nov. isolated from California sea lions (Zalophus californianus). Int J Syst Evol Microbiol. https://doi.org/10.1099/ijsem.0.003729(Epub ahead of print)
Yavari CA, Ramirez AS, Nicholas RAJ, Radford AD, Darby AC, Bradbury JM (2017) Mycoplasma tullyi sp. nov., isolated from penguins of the genus Spheniscus. Int J Syst Evol Microbiol 67:3692–3698
Zavala A, Naya H, Romero H, Sabbia V, Piovani R, Musto H (2005) Genomic GC content prediction in prokaryotes from a sample of genes. Gene 357:137–143
Ziegler L, Moller Palau-Ribes F, Enderlein D, Herbst W, Schmidt L, Lierz M (2019) Mycoplasma hafezii sp. nov., isolated from the trachea of a peregrine falcon (Falco peregrinus). Int J Syst Evol Microbiol 69:773–777
We dedicate this study in memory of Drs. Ruth M. Lemcke and H. Louise Ruhnke for their outstanding scientific contributions to Mycoplasmology research focused on studying animal mycoplasmas, and for the primary isolation of Acholeplasma equirhinis sp. nov. and Mycoplasma procyoni sp. nov., respectively, which were deposited by them to the International Organization for Mycoplasmology Mollicutes Collection and thus saved these unique microorganisms for future characterization and scientific studies.
The authors received no specific funds from any funding agency.
Conflict of interest
The authors declare no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Maureen K. Davidson and Vladimir E. Chizhikov currently retired from the US FDA.
The GenBank accession number for the 16S rRNA gene sequences of the type strains N93T and LR5794T are EU925159 and FJ226566, respectively, and the GenBank accession numbers for other housekeeping genes used for the genetic characterization of each novel species described in this study are provided in the manuscript.
Communicated by Erko Stackebrandt.
About this article
Cite this article
Volokhov, D.V., Gao, Y., Davidson, M.K. et al. Acholeplasma equirhinis sp. nov. isolated from respiratory tract of horse (Equus caballus) and Mycoplasma procyoni sp. nov. isolated from oral cavity of raccoon (Procyon lotor). Arch Microbiol 202, 411–420 (2020). https://doi.org/10.1007/s00203-019-01786-x