scholarly journals Phylogeny of the Mycoplasma mycoides cluster based on analysis of five conserved protein-coding sequences and possible implications for the taxonomy of the group

2007 ◽  
Vol 57 (10) ◽  
pp. 2247-2258 ◽  
Author(s):  
Lucía Manso-Silván ◽  
Xavier Perrier ◽  
François Thiaucourt
Keyword(s):  
Housekeeping Genes ◽  
New Techniques ◽  
Protein Coding ◽  
Coding Sequences ◽  
Individual Gene ◽  
Mycoplasma Capricolum ◽  
Large Colony ◽  
Mycoplasma Mycoides ◽  
Small Colony ◽  
Distinct Branch

A phylogenetic tree of the Mycoplasma mycoides cluster was inferred from a set of concatenated sequences from five housekeeping genes (fusA, glpQ, gyrB, lepA and rpoB). The relevance of this phylogeny was reinforced by detailed analysis of the congruence of the phylogenies derived from each of the five individual gene sequences. Two subclusters were distinguished. The M. mycoides subcluster comprised M. mycoides subsp. mycoides biotypes Small Colony (SC) and Large Colony (LC) and M. mycoides subsp. capri. The latter two groups could not be clearly separated, which supports previous proposals that they be united into a single taxonomic entity. The Mycoplasma capricolum subcluster included M. capricolum subsp. capricolum, M. capricolum subsp. capripneumoniae and Mycoplasma sp. bovine group 7 of Leach, a group of strains that remains unassigned. This group constituted a distinct branch within this cluster, supporting its classification as a subspecies of M. capricolum. Mycoplasma cottewii and Mycoplasma yeatsii clustered in a group that was distinct from Mycoplasma putrefaciens and they were all clearly separated from the M. mycoides cluster. In conclusion, this approach has allowed us to assign phylogenetic positions to all members of the M. mycoides cluster and related species and has proved the need to adjust the existing taxonomy. Furthermore, this method may be used as a reference technique to assign an unequivocal position to any particular strain related to this cluster and may lead to the development of new techniques for rapid species identification.

scholarly journals Variable Surface Protein Vmm of Mycoplasma mycoides subsp. mycoides Small Colony Type

2002 ◽  
Vol 184 (13) ◽  
pp. 3712-3722 ◽  
Author(s):  
Anja Persson ◽  
Karin Jacobsson ◽  
Lars Frykberg ◽  
Karl-Erik Johansson ◽  
François Poumarat
Keyword(s):  
Phase Variation ◽  
Surface Protein ◽  
Transcriptional Level ◽  
Colony Type ◽  
Mycoplasma Capricolum ◽  
Binding Epitope ◽  
Variable Surface ◽  
Mycoplasma Mycoides ◽  
Small Colony ◽  
Reversible Phase

ABSTRACT A variable surface protein, Vmm, of the bovine pathogen Mycoplasma mycoides subsp. mycoides small colony type (M. mycoides SC) has been identified and characterized. Vmm was specific for the SC biotype and was expressed by 68 of 69 analyzed M. mycoides SC strains. The protein was found to undergo reversible phase variation at a frequency of 9 × 10−4 to 5 × 10−5 per cell per generation. The vmm gene was present in all of the 69 tested M. mycoides SC strains and encodes a lipoprotein precursor of 59 amino acids (aa), where the mature protein was predicted to be 36 aa and was anchored to the membrane by only the lipid moiety, as no transmembrane region could be identified. DNA sequencing of the vmm gene region from ON and OFF clones showed that the expression of Vmm was regulated at the transcriptional level by dinucleotide insertions or deletions in a repetitive region of the promoter spacer. Vmm-like genes were also found in four closely related mycoplasmas, Mycoplasma capricolum subsp. capricolum, M. capricolum subsp . capripneumoniae, Mycoplasma sp. bovine serogroup 7, and Mycoplasma putrefaciens. However, Vmm could not be detected in whole-cell lysates of these species, suggesting that the proteins encoded by the vmm-like genes lack the binding epitope for the monoclonal antibody used in this study or, alternatively, that the Vmm-like proteins were not expressed.

scholarly journals The ability of Mycoplasma mycoides subspecies mycoides and closely related strains from goats and sheep to immunize mice against subspecies capri

1981 ◽  
Vol 87 (2) ◽  
pp. 321-329 ◽  
Author(s):  
G. R. Smith ◽  
Janet C. Oliphant
Keyword(s):  
Cross Protection ◽  
Contagious Bovine Pleuropneumonia ◽  
Large Colony ◽  
Mycoplasma Mycoides ◽  
Small Colony

SummarySmall colony (SC) strains of Mycoplasma mycoides subsp. mycoides from contagious bovine pleuropneumonia (CBPP) and from goats were compared with large colony (LC) strains of so-called M. mycoides subsp. mycoides from goats and sheep by means of a cross-protection test in which mice were challenged with M. mycoides subsp. capri.Of 13 LC strains, all gave partial cross-protection, and 11 were shown to be more closely related than four SC strains to subspecies capri. In a further experiment, six SC strains – three from CBPP and three from goats – all gave weak partial cross-protection against subspecies capri.

scholarly journals Versatile Use of oriC Plasmids for Functional Genomics of Mycoplasma capricolum subsp. capricolum

2005 ◽  
Vol 71 (6) ◽  
pp. 2888-2893 ◽  
Author(s):  
Carole Janis ◽  
Carole Lartigue ◽  
Joachim Frey ◽  
Henri Wróblewski ◽  
François Thiaucourt ◽  
...  
Keyword(s):  
Homologous Recombination ◽  
Functional Genomics ◽  
Gene Inactivation ◽  
Heterologous Proteins ◽  
Gene Encoding ◽  
Colony Type ◽  
Genome Data ◽  
Mycoplasma Capricolum ◽  
Large Colony ◽  
Mycoplasma Mycoides

ABSTRACT Replicative oriC plasmids were recently developed for several mollicutes, including three Mycoplasma species belonging to the mycoides cluster that are responsible for bovine and caprine diseases: Mycoplasma mycoides subsp. mycoides small-colony type, Mycoplasma mycoides subsp. mycoides large-colony type, and Mycoplasma capricolum subsp. capricolum. In this study, oriC plasmids were evaluated in M. capricolum subsp. capricolum as genetic tools for (i) expression of heterologous proteins and (ii) gene inactivation by homologous recombination. The reporter gene lacZ, encoding β-galactosidase, and the gene encoding spiralin, an abundant surface lipoprotein of the related mollicute Spiroplasma citri, were successfully expressed. Functional Escherichia coli β-galactosidase was detected in transformed Mycoplasma capricolum subsp. capricolum cells despite noticeable codon usage differences. The expression of spiralin in M. capricolum subsp. capricolum was assessed by colony and Western blotting. Accessibility of this protein at the cell surface and its partition into the Triton X-114 detergent phase suggest a correct maturation of the spiralin precursor. The expression of a heterologous lipoprotein in a mycoplasma raises potentially interesting applications, e.g., the use of these bacteria as live vaccines. Targeted inactivation of gene lppA encoding lipoprotein A was achieved in M. capricolum subsp. capricolum with plasmids harboring a replication origin derived from S. citri. Our results suggest that the selection of the infrequent events of homologous recombination could be enhanced by the use of oriC plasmids derived from related mollicute species. Mycoplasma gene inactivation opens the way to functional genomics in a group of bacteria for which a large wealth of genome data are already available and steadily growing.

scholarly journals Plasmids in Mycoplasma species isolated from goats and sheep and their preliminary typing

1999 ◽  
Vol 30 (1) ◽  
pp. 32-36 ◽  
Author(s):  
Elmiro R. Nascimento ◽  
Al J. DaMassa ◽  
Richard Yamamoto ◽  
M. Graça F. Nascimento
Keyword(s):  
Enzyme Digestion ◽  
Mycoplasma Species ◽  
Restriction Enzyme Digestion ◽  
Extrachromosomal Dna ◽  
Dna Virus ◽  
Clinically Normal ◽  
Mycoplasma Capricolum ◽  
Large Colony ◽  
Mycoplasma Mycoides ◽  
Dna Elements

One-hundred-five (105) clinical isolates of mycoplasma from caprine origin and one isolate from ovine were surveyed for plasmids, which were present in thirty-three (31%) of them. These mycoplasmas originated from 13 herds. Ten of them were symptomatic for mycoplasmal disease (mastitis, polyarthritis, septicemia) and three herds were asymptomatic, i.e., clinically normal. Twenty-eight isolates were Mycoplasma mycoides subspecies mycoides LC (large colony or caprine biotype), four were Mycoplasma capricolum subsp. capricolum and one was Mycoplasma cottewii. The isolated plasmids were linearized by EcoRI, EcoRV, EcoRI and EcoRV or BamHI and EcoRV, and were of five sizes (1.1, 1.6, 1.7, 1.8, and 1.9 Kbp). Based on restriction enzyme digestion and size of the linearized supercoiled extrachromosomal DNA, five plasmid types were recovered (p1II, p2III, p2V, p3I, and p4IV). The small size of these DNA elements probably exclude replicative forms of DNA virus, which are equal or larger than 8.0 Kbp.

scholarly journals Flow Cytometric Determination of the Effects of Antibacterial Agents on Mycoplasma agalactiae, Mycoplasma putrefaciens, Mycoplasma capricolum subsp. capricolum, and Mycoplasma mycoides subsp. mycoides Large Colony Type

2006 ◽  
Vol 50 (8) ◽  
pp. 2845-2849 ◽  
Author(s):  
Patricia Assunção ◽  
Nuno T. Antunes ◽  
Ruben S. Rosales ◽  
Carlos Poveda ◽  
Jose B. Poveda ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Propidium Iodide ◽  
Antibacterial Agents ◽  
Mycoplasma Species ◽  
Colony Type ◽  
Flow Cytometric ◽  
Mycoplasma Capricolum ◽  
Large Colony ◽  
Mycoplasma Mycoides

ABSTRACT Flow cytometry together with SYBR green I and propidium iodide was used to study the effects of enrofloxacin, ciprofloxacin, gentamicin, chloramphenicol, oxytetracycline, and tylosin on four mycoplasma species. Inhibition of mycoplasma growth could be detected by as early as 3 h after the start of treatment. The strongest effect was observed with enrofloxacin- and ciprofloxacin-treated cells.

scholarly journals Further studies on caprine and ovine mycoplasmas related to Mycoplasma mycoides subsp. mycoides

1980 ◽  
Vol 85 (2) ◽  
pp. 247-256 ◽  
Author(s):  
G. R. Smith ◽  
J. M. Hooker ◽  
R. A. Milligan
Keyword(s):  
Cross Protection ◽  
Causative Organism ◽  
Complete Protection ◽  
Challenge Strain ◽  
Barbary Sheep ◽  
Large Colony ◽  
Mycoplasma Mycoides ◽  
Homologous Strain ◽  
Small Colony

SUMMARYNine caprine and ovine mycoplasma strains, said to be indistinguishable serologically from Mycoplasma mycoides subsp. mycoides (the causative organism of contagious bovine plouropneumonia; CBPP) were examined in mice by (1) a mycoplasmaemia test, and (2) a cross-protection test. Of the nine strains, two from goats belonged to a small colony (SC) type; four caprine and three ovine strains belonged to a large colony (LC) type.The two SC strains – like a single SC strain examined in an earlier study – were indistinguishable from genuine M. mycoides subsp. mycoidesas isolated from CBPP. They produced mycoplasmaemia readily. In a cross-protection test, the two SC strains and a CBPP strain immunized completely against each other.Of the seven LC strains, six – like six LC strains examined in an earlier study – were easily distinguished from genuine M. mycoides subsp. mycoides; except for one that was not tested, all were shown to lack the ability to produce mycoplasmaemia readily. In cross-protection tests all six strains immunized partially but not completely against a CBPP strain.The soventh LC strain (Mankefár 2833) was exceptional: it produced mycoplasmaemia readily, resembling tho SC strains in this respect. Like other LC strains, in cross-protection tests it protected only partially against a CBPP strain. Strain Mankef´r 2833 was isolated in ca. 1005 by Brack from a Barbary sheep (Ammolragus lervia) in a German zoo.The ability of Mankefár 2833 to produce mycoplasmaemia enabled it to be used as a challenge strain in cross-protection tests. For the purpose of such tests the collection of nine mycoplasma strains referred to above was augmented with six LC strains from an earlier study. Partial but not complete protection against Mankefár 2833 was produced by two caprine SC strains, one CBPP strain, and nine LC strains. Three further LC strains gave protection that may have been as strong as that produced by the homologous strain, but confirmatory experiments are needed. A strain of M. mycoides subsp. capri gave no protection against Mankefár 2833.

scholarly journals A study of F38-type and related mycoplasmas by mycoplasmaemia and cross-immunization tests in mice

1984 ◽  
Vol 93 (3) ◽  
pp. 465-473 ◽  
Author(s):  
M. Kanyi Kibe ◽  
G. R. Smith
Keyword(s):  
Cross Protection ◽  
Partial Protection ◽  
Colony Type ◽  
Mycoplasma Capricolum ◽  
Large Colony ◽  
Strain Nctc ◽  
History Of ◽  
Small Colony ◽  
Type Strains

SummaryIn vivo methods were used to study the F38-type mycoplasma in parallel with related mycoplasmas.Three of five strains of ‘bovine serogroup 7’ with an unknown history of subculture produced mycoplasmaemia in mice inoculated intraperitoneally. A strain of ‘bovine serogroup L’ also produced mycoplasmaemia, but no evidence of similar ability could be found for single strains of Mycoplasma capricolum, M. equigenitalium and M. primatum, or for two strains of the F38-type mycoplasma.In cross-immunization tests a bovine serogroup 7 strain (NCTC 10133) and a strain (‘Blenheim’) of the SC (small colony) type of M. mycoides subsp. mycoides were used for the purpose of challenge. Cross-protection was described as ‘complete’ or ‘partial’, depending on whether it was as great as, or less than, that produced by homologous vaccine. Although strain NCTC 10133 protected strongly, possibly completely, against Blenheim, and Blenheim gave partial protection against NCTC 10133, challenge with NCTC 10133 and Blenheim gave strikingly different results. Thus (1) F38-type strains, M equigenitalium and M. primatum all gave partial cross-protection against NCTC 10133 but not against Blenheim, (2) NCTC 10133, unlike Blenheim, was seldom susceptible to partial cross-protection by LC (large colony) strains of M. mycoides subsp. mycoides, and (3) three SC strains – which would have protected completely against Blenheim – protected only partially against NCTC 10133. NCTC 10133 and Blenheim were similar, however, in that M. capricolum and M. mycoides subsp. capri failed to cross-protect against them both.

scholarly journals Screening of Hungarian Cattle Herds for Mycoplasma mycoides Subspecies Mycoides Small Colony Infection with Negative Results

2000 ◽  
Vol 48 (4) ◽  
pp. 375-385
Author(s):  
L. Stipkovits ◽  
Á Dán ◽  
Erika Varga ◽  
Paula De Santis ◽  
Rosella Lelly ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Specific Primers ◽  
Serum Dilution ◽  
Negative Results ◽  
Large Colony ◽  
Nasal Swabs ◽  
Mycoplasma Mycoides ◽  
Small Colony ◽  
Polymerase Chain ◽  
Cattle Herds

At abattoirs and farms, 1248 sera were collected from animals representing 121 farms, and examined by complement fixation test using Mycoplasma mycoides subspecies mycoides small colony type (MmmSC) antigen. All sera were negative except seven from four farms, giving ++ reactions in the serum dilution of 1:10. On retesting, these sera and additional 30 sera collected repeatedly in both farms gave negative results. In isolation attempts, 953 lung samples collected from slaughtered cattle at the same abattoirs, and 326 nasal swabs collected from 11 herds proved to be negative for the presence of MmmSC, but M. bovis was isolated frequently. In the small farms 23.95% of the animals had pleurisy and/or pneumonia while in the large herds 34.69% had lesions. DNA extracted from 50 nasal swabs and 430 lung samples was examined by polymerase chain reaction (PCR) using M. mycoides cluster-specific primers. DNA from further 325 lung samples was tested by the more specific M. mycoides subspecies mycoides small colony/large colony/capri specific primers and 196 samples by nested PCR specific for MmmSC. All gave negative results. The detection level of cluster-specific primers and the more specific primers was 33.4 pg of DNA, whereas that of nested PCR was 0.33 pg.

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