An Alaska Native hunter developed fever, swollen finger, and septic hips after harvesting seals. Evaluation of hip tissue by 16S rRNA gene polymerase chain reaction and sequencing revealed a putative novel mycoplasma species. We report the identification of this organism and describe the first known case of disseminated seal finger mycoplasmosis.

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A hybrid sequence assembly of the complete Mycoplasma synoviae type strain WVU 1853T genome was compared to that of strain MS53. The findings support prior conclusions about M. synoviae, based on the genome of that otherwise uncharacterized field strain, and provide the first evidence of epigenetic modifications in M. synoviae.

M ycoplasma canis infects many mammalian hosts but is usually thought of as a commensal or opportunistic cofactor in respiratory or urogenital tract diseases of dogs (1). We found unexpectedly that M. canis was also detectable by culture or PCR in a majority of brain tissue specimens in a retrospective case-control study of canine granulomatous meningoencephalitis (ME) (GME) and necrotizing ME (NME) (2). The presence of M. canis in brain tissue was associated with both GME and NME (both P Ͻ 0.05, as determined by a 2 test). The clinical signs of this common idiopathic neurological disease of dogs include seizures, proprioceptive deficits, circling, and blindness. Immunosuppressive therapy may be palliative, but the syndrome is progressive and uniformly fatal (3). The extensive search for a presumed viral cause of canine GME and NME has been fruitless (4).In humans, bacterial meningitis and encephalitis are multifactorial lethal infections with often severe sequelae for survivors. New detection methods have shown that the variety of bacteria associated with human ME is much more extensive than usually appreciated (5-9). Additional animal models of bacterial ME are necessary to study this broader spectrum of pathogens (10). Since a possible association between M. canis and canine ME was discovered, our objective has been to help fill the void of basic knowledge about the organism's virulence factors, the host responses that it elicits, and its potential roles in pathogenesis. Our working hypotheses were that M. canis is capable of evoking host cell responses that favor dissemination from mucosal surfaces to secondary sites of infection, possibly in a strain-dependent fashion, and also that, regardless of how it might reach those sites, the presence of M. canis there modulates inflammation and direct injury to host cells. Understanding this potential can be expected to help evaluate the cause of canine ME and other diseases.(Portions of these data were presented in abstract form at Congresses of the International Organization for Mycoplasmology [90,91].) MATERIALS AND METHODSMycoplasma strains and cultivation. Strain PG14 T of M. canis (ATCC 19525) was first isolated from the throat of a normal dog (11). Strains UF31, UF33, LV, 5, 26, Cal, and Mara were first isolated from vaginal swabs of dogs without ME (12). Strains UFG1, UFG2, UFG3, and UFG4 were isolated from frozen brain tissues from cases of canine NME (2). Mycoplasma cynos strain H-831 T (ATCC 27544) was first isolated from the lung of a dog with pneumonia (13). Mycoplasma arginini strain G230 T , Mycoplasma bovigenitalium strain PG11 T , Mycoplasma edwardii strain PG24 T , Mycoplasma maculosum strain Skotti B, Mycoplasma molare strain H542 T , Mycoplasma opalescens strain MH5408 T , and Mycoplasma spumans strain PG13T , representing other species that have been isolated from dogs (1), were obtained from The Mollicutes Collection. All strains were propagated under standard conditions (14) in ATCC 988 medium supplemented with fetal bovine serum (FBS) and glu...

bTo understand its potential to cause invasive disease, the genome of Mycoplasma canis strain PG14T from a dog's throat was compared to those of isolates from the genital tract or brain of dogs. The average nucleotide identity between strain pairs is 98%, and their genome annotations are similar. Mycoplasma canis is a mucosal surface commensal or opportunistic pathogen (4,5,6). To understand its potential to cause invasive disease (2), we compared genomes of isolates from the genital tract (UF31 and UF33) or brain (UFG1 and UFG4) of dogs to strain PG14T from a dog's throat. DNA sequences obtained through multiplex Illumina GAIIx paired-end sequencing (mean of 10,226,327 quality-filtered 101-nucleotide [nt] reads per strain, yielding coverage depth of Ͼ1,000ϫ each) were assembled with Galaxy (1), CLC Genomics Workbench v4.8, and Sequencher v4.8 software. The numbers of contigs of Ͼ500 bp obtained by de novo assembly were 62, 30, 41, 19, and 25 for strains UF31, UF33, UFG1, UFG4, and PG14 T , respectively. Open reading frames (ORFs) were predicted using GeneMark, Glimmer, and Prodigal hidden Markov model (HMM)-based models plus tRNAscan-SE. Annotation was by BLAST comparison to NCBI's Protein Clusters database, all proteins from complete microbial genomes, and the Rfam noncoding RNA database. The average nucleotide identity between strains was calculated using JSpecies v1.2.1 (11).The noncontiguous finished draft M. canis PG14 T genome includes 867,208 nt in 6 contigs of Ͼ21 kb plus 27,894 nt in contigs of Ͻ10 kb, an overall size which is substantially larger than the 795 kb estimated by pulsed-field gel electrophoresis (PFGE) (4). Its GϩC content is 27%, less than the 28.4% and 29.1% estimated by thermal denaturation and buoyant density, respectively (8, 10). Of 696 predicted ORFs, about three-fourths have assigned functions, including 31 structural RNAs, 31 proteins involved in metabolism of amino acids, purines, pyrimidines, nucleosides, nucleotides, cofactors, prosthetic groups, or carriers, 7 proteins involved in fatty acid or phospholipid metabolism, 75 proteins involved in intermediary or energy metabolism, 71 proteins involved in transport or binding, 257 proteins involved in nucleic acid metabolism, transcription, or translation or protein fate, 10 proteins involved in regulatory functions, including HrcA, ArsR, GntR, LacI, and RpiR family transcription regulators, 12 proteins involved in cell division, 20 cell envelope lipoproteins, and 1 IS256-type transposase. The proportions of the genome assigned to various role categories are similar to those of related species in the Mycoplasma synoviae phylogenetic cluster.M. canis encodes the tyrosine recombinase XerC but no recognized variable surface antigens or adhesins (7). Virulence factor candidates include a Fic family AMPylator (12), a putative secreted effector protein with an Arg-Gly-Asp host celltargeting motif, a predicted 635-kDa protein with multiple immunoglobulin/albumin-binding domains, and the secreted glycosidases sialidase (but no path for sia...

The genome of Mycoplasma phocidae strain 105T was analyzed in order to improve our understanding of its role in epidemic marine mammal mortalities. It was found to encode a suite of immunosuppressors that may enable evasion of host defenses and modulate susceptibility to viral coinfections or their severity in seals.

The genome of Mycoplasma phocirhinis strain 852T was examined for determinants of tropism or virulence. It encodes multiple orthologs of an immunosuppressor that may predispose susceptibility to infection or influence outcomes of intercurrent diseases in marine mammals.

The Mycoplasma phocicerebrale genome was analyzed to better understand this opportunistic pathogen. Amplification with ϕ29 polymerase was used to generate enough genomic DNA for large-insert library construction. Like other mycoplasmas from seals, M. phocicerebrale encodes an immunosuppressor that may predispose susceptibility to infection or influence intercurrent diseases of affected hosts.