Mycoplasma species bovine group 7, represented by the type strain PG50, is one of six members of the Mycoplasma mycoides cluster and has been implicated in sporadic and outbreak cases of polyarthritis and mastitis in Australian dairy cattle. This study describes cloning and sequencing a 7·9 kb region of the PG50 chromosome and identification of genes involved in glycerol transport (gtsA, gtsB and gtsC) that are followed by a putative lipoprotein gene lppB and a genomic locus containing two ORFs encoding putative membrane proteins. Long range PCR using primers spanning gtsABC and downstream flanking genes, and Southern hybridization analyses using a suite of probes derived from M. mycoides subsp. mycoides small colony type (SC) strain Afadé for gtsA, gtsB and gtsC, lppB and the two downstream genes confirmed that these genes were conserved among Mycoplasma sp. bovine group 7 isolates and mycoplasmas belonging to the M. mycoides subcluster [M. mycoides subsp. mycoides SC, M. mycoides subsp. mycoides large colony type (LC) and M. mycoides subsp. capri] but were absent in mycoplasmas belonging to the Mycoplasmacapricolum subcluster (M. capricolum subsp. capricolum and M. capricolum subsp. capripneumoniae). M. capricolum subsp. capricolum type strain California kid did not hybridize with the probe for gtsA and gave only weak or no hybridization signals with probes derived from the loci downstream of gtsABC, suggesting that this region has diverged in mycoplasmas belonging to subspecies of M. capricolum. It is shown that PG50, after the addition of a physiological concentration of glycerol to the growth medium, generates H2O2 at levels comparable with strain Afadé, implying that the glycerol transport system is functional in Mycoplasma sp. bovine group 7. This suggests that in PG50, as in M. mycoides subsp. mycoides SC, glycerol uptake is followed by phosphorylation to glycerol 3-phosphate and then conversion to dihydroxyacetone phosphate, catalysed by l-α-glycerophosphate oxidase, resulting in the production of H2O2. The ability of Mycoplasma sp. bovine group 7 to generate significant amounts of hydrogen peroxide may be important in pathogenesis.
AlexanderP. G,
SleeK. J,
McOristS,
IrelandL.,
ColoeP. J.
1985; Mastitis in cows and polyarthritis and pneumonia in calves caused by Mycoplasma species bovine group 7. Aust Vet J 62:135–136
BölskeG,
MsamiH,
HumlesjöN. E,
ErnøH.,
JönssonL.
1988; Mycoplasma capricolum in an outbreak of polyarthritis and pneumonia in goats. Acta Vet Scand 29:331–338
BonnetF,
SaillardC,
BovéJ. M,
LeachR. H,
RoseD. L,
CottewG. S.,
TullyJ. G.
1993; DNA relatedness between field isolates of Mycoplasma F38-group, the agent of contagious caprine pleuropneumonia, and strains of Mycoplasma capricolum
. Int J Syst Bacteriol 43:597–602
ChambaudI,
HeiligR,
FerrisS.9 other authors2001; The complete genome sequence of the murine respiratory pathogen Mycoplasma pulmonis
. Nucleic Acids Res 29:2145–2153
ChengX,
NicoletJ,
PoumaratF,
RegallaJ,
ThiaucourtF.,
FreyJ.
1995; Insertion element IS 1296 in Mycoplasma mycoides subsp. mycoides small colony identifies a European clonal line distinct from the African and Australian strains. Microbiology 141:3221–3228
ChristiansenC.,
ErnøH.
1990; RFLP in rRNA genes of Mycoplasma capricolum , the caprine F38-like group and the bovine serogroup 7. Zentbl Bakteriol 20:479–488
CostasM,
LeachR. H.,
MitchelmoreD. L.
1987; Numerical analysis of PAGE protein patterns and the taxonomic relationships within the ‘ Mycoplasma mycoides cluster’. J Gen Microbiol 133:3319–3329
DjordjevicS. P,
ForbesW. A,
Forbes-FaulknerJ,
KuhnertP,
HumS,
HornitzkyM. A,
VileiE. M.,
FreyJ.
2001; Genetic diversity among Mycoplasma species bovine group 7: clonal isolates from an outbreak of polyarthritis, mastitis, and abortion in dairy cattle. Electrophoresis 22:3551–3561
FreyJ,
ChengX,
KuhnertP.,
NicoletJ.
1995; Identification and characterisation of IS 1296 in Mycoplasma mycoides subsp. mycoides SC and presence in related mycoplasmas. Gene 160:95–100
FreyJ,
ChengX,
MonneratM.-P,
AbdoE.-M,
KrawinklerM,
BölskeG.,
NicoletJ.
1998; Genetic and serological analysis of the immunogenic 67-kDa lipoprotein of Mycoplasma sp. bovine group 7. Res Microbiol 149:55–64
GonçalvesR,
RegallaJ,
NicoletJ,
FreyJ,
NicholasR,
BashiruddinJ,
De SantisP.,
GonçalvesA. P.
1998; Antigen heterogeneity among Mycoplasma mycoides subsp. mycoides SC isolates: discrimination of major surface proteins. Vet Microbiol 63:13–28
GuerinC,
ThiaucourtF,
MadyV,
BréardA.,
LefèvreP. C.
1993; Rapid diagnosis of contagious caprine pleuropneumonia in pleural fluids by immunoblotting assay. Small Rumin Res 12:193–200
HimmelreichR,
HilbertH,
PlagensH,
PirklE,
LiB. C.,
HerrmannR.
1996; Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae
. Nucleic Acids Res 24:4420–4449
HoushaymiB. M,
MilesR. J.,
NicholasR. A.
1997; Oxidation of glycerol differentiates African from European isolates of Mycoplasma mycoides subspecies mycoides SC (small colony. Vet Rec 140:182–183
HumS,
KessellA,
DjordjevicS. P,
RheinbergerR,
HorntizkyM,
ForbesW.,
GonsalvesJ.
2000; Mastitis, polyarthritis and abortion caused by Mycoplasma species bovine group 7 in dairy cattle. Aust Vet J 78:744–750
KibeM. K,
BidwellD. E,
TurpP.,
SmithG. R.
1985; Demonstration of cross-reactive antigens in F38 and related mycoplasmas by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. J Hyg 95:95–106
MonneratM.-P,
ThiaucourtF,
NicoletJ.,
FreyJ.
1999; Comparative analysis of the lppA locus in Mycoplasma capricolum subsp.capricolum and Mycoplasma capricolum subsp. capripneumoniae
. Vet Microbiol 69:157–172
OlssonB,
BölskeG,
BergströmK.,
JohanssonK.-E.
1990; Analysis of caprine mycoplasmas and mycoplasma infections in goats using two-dimensional electrophoresis and immunoblotting. Electrophoresis 11:861–869
RiceP,
HoushaymiB. M,
NicholasR. A.,
MilesR. J.
2000; A rapid biochemical test to aid identification of Mycoplasma mycoides subspecies mycoides small colony (SC) strains. Lett Appl Microbiol 30:70–74
RodwellA. W.,
RodwellE. S.
1978; Relationships between strains of Mycoplasma mycoides subsp. mycoides and capri studied by two-dimensional gel electrophoresis of cell proteins. J Gen Microbiol 109:259–263
ThiaucourtF,
BölskeG,
LibeauG,
LeGoffC.,
LefèvreP. C.
1994; The use of monoclonal antibodies in the diagnosis of contagious caprine pleuropneumonia (CCCP). Vet Microbiol 41:191–203
VileiE. M.,
FreyJ.
2001; Genetic and biochemical characterization of glycerol uptake in Mycoplasma mycoides subsp. mycoides SC: its impact on H2O2 production and virulence. Clin Diagn Lab Immunol 8:85–92
VileiE. M,
NicoletJ.,
FreyJ.
1999; IS 1634 , a novel insertion element creating long, variable-length direct repeats which is specific for Mycoplasma mycoides subsp. mycoides small-colony type. J Bacteriol 181:1319–1323
VileiE. M,
AbdoE.-M,
NicoletJ,
BotelhoA,
GonçalvesR.,
FreyJ.
2000; Genomic and antigenic differences between the European and African/Australian clusters of Mycoplasma mycoides subsp. mycoides SC. Microbiology 146:477–486