Characterization of Strains of Mycoplasma mycoidessubsp. mycoides Small Colony Type Isolated from Recent Outbreaks of Contagious Bovine Pleuropneumonia in Botswana and Tanzania:  Evidence for a New Biotype

Four strains of Mycoplasma mycoides subsp.mycoides small colony type (MmmSC) isolated from recent outbreaks of contagious bovine pleuropneumonia (CBPP) in Africa have been investigated. One Botswanan strain, M375, displayed numerous and significant phenotypic differences from both contemporary field isolates and older field and vaccine strains (African, Australian, and European strains dating back to 1936). Differences include altered morphology, reduced capsular polysaccharide production, high sensitivity to MmmSC rabbit hyperimmune antisera in vitro, and unique polymorphisms following immunoblotting. While insertion sequence analysis using IS1634 clearly indicates a close evolutionary relationship to west African strains, hybridization with IS1296 shows the absence of a band present in all other strains of MmmSC examined. The data suggest that a deletion has occurred in strain M375, which may explain its altered phenotype, including poor growth in vitro and a relative inability to cause septicemia in mice. These characteristics are also exhibited byMycoplasma capricolum subsp. capripneumoniae(causal agent of contagious caprine pleuropneumonia [CCPP]), against which M375 antiserum exhibited some activity in vitro (unique among the various MmmSC antisera tested). These findings may have evolutionary implications, since CCPP is believed to be lung specific and without a septicemic phase (unlike CBPP). Since M375 was isolated from a clinical case of CBPP, this novel biotype may be fairly widespread but not normally isolated due to difficulty of culture and/or a potentially altered disease syndrome. Bovine convalescent antisera (obtained from contemporary naturally infected cattle in Botswana) were active against strain M375 in an in vitro growth inhibition test but not against any other strains of MmmSC tested. There exists the possibility therefore, that strain M375 may possess a set of protective antigens different from those of other strains of MmmSC (including vaccine strains). These findings have implications for the control of the current CBPP epidemic in Africa.

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Assessing the In Vitro Effectiveness of Antimicrobials against Mycoplasma mycoides subsp. mycoides Small-Colony Type To Reduce Contagious Bovine Pleuropneumonia Infection

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.49.12.5162-5165.2005 ◽

2005 ◽

Vol 49 (12) ◽

pp. 5162-5165 ◽

Cited By ~ 11

Author(s):

R. D. Ayling ◽

S. Bisgaard-Frantzen ◽

J. B. March ◽

K. Godinho ◽

R. A. J. Nicholas

Keyword(s):

Antimicrobial Resistance ◽

Contagious Bovine Pleuropneumonia ◽

Colony Type ◽

Minimum Inhibitory Concentrations ◽

Mycoplasma Mycoides ◽

Small Colony

ABSTRACT In vitro minimum inhibitory concentrations were determined for 21 antimicrobials against 41 isolates of Mycoplasma mycoides subsp. mycoides small-colony type, the cause of contagious bovine pleuropneumonia. Of the antimicrobials used most widely in Africa, oxytetracycline and tilmicosin were effective, while the isolates were resistant to tylosin. These results provide a baseline for monitoring antimicrobial resistance.

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Cytotoxicity of Mycoplasma mycoides subsp. mycoides Small Colony Type to Bovine Epithelial Cells

Infection and Immunity ◽

10.1128/iai.00938-07 ◽

2007 ◽

Vol 76 (1) ◽

pp. 263-269 ◽

Cited By ~ 35

Author(s):

Daniela F. Bischof ◽

Carole Janis ◽

Edy M. Vilei ◽

Giuseppe Bertoni ◽

Joachim Frey

Keyword(s):

Close Contact ◽

Clinical Signs ◽

Host Cells ◽

Uptake System ◽

Contagious Bovine Pleuropneumonia ◽

Colony Type ◽

Mycoplasma Mycoides ◽

Small Colony ◽

High Cytotoxicity

ABSTRACT The cytotoxicities of various strains of Mycoplasma mycoides subsp. mycoides small colony type (SC), the agent of contagious bovine pleuropneumonia (CBPP), were measured in vitro using embryonic calf nasal epithelial (ECaNEp) cells. Strains isolated from acute cases of CBPP induced high cytotoxicity in the presence of glycerol, concomitant with the release of large amounts of toxic H2O2 that were found to be translocated into the cytoplasms of the host cells by close contact of the Mycoplasma strains with the host cells. Currently used vaccine strains also showed high cytotoxicity and high H2O2 release, indicating that they are attenuated in another virulence attribute. Strains isolated from recent European outbreaks of CBPP with mild clinical signs, which are characterized by a defect in the glycerol uptake system, released small amounts of H2O2 and showed low cytotoxicity to ECaNEp cells. M. mycoides subsp. mycoides SC strain PG1 released large amounts of H2O2 but was only slightly cytotoxic. PG1 was found to have a reduced capacity to bind to ECaNEp cells and was unable to translocate H2O2 into the bovine cells, in contrast to virulent strains that release large amounts of H2O2. Thus, an efficient translocation of H2O2 into host cells is a prerequisite for the cytotoxic effect and requires an intact adhesion mechanism to ensure a close contact between mycoplasmas and host cells.

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Comparison of in vitro activity of danofloxacin, florlenicol, oxytetracycline, spectinomycin and tilmicosin against Mycoplasma mycoides subspecies mycoides small colony type

Veterinary Record ◽

10.1136/vr.146.9.243 ◽

2000 ◽

Vol 146 (9) ◽

pp. 243-246 ◽

Cited By ~ 14

Author(s):

R. D. Ayling ◽

S. E. Baker ◽

R. A. J. Nicholas ◽

M. L. Peek ◽

A. J. Simon

Keyword(s):

Vitro Activity ◽

In Vitro Activity ◽

Colony Type ◽

Mycoplasma Mycoides ◽

Small Colony

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Assessment of in vitro interferon-γ responses from peripheral blood mononuclear cells of cattle infected with Mycoplasma mycoides ssp. mycoides small colony type

Veterinary Immunology and Immunopathology ◽

10.1016/j.vetimm.2008.02.019 ◽

2008 ◽

Vol 124 (1-2) ◽

pp. 192-197 ◽

Cited By ~ 14

Author(s):

Joerg Jores ◽

Isabel Nkando ◽

Anja Sterner-Kock ◽

Wolfram Haider ◽

Jane Poole ◽

...

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Interferon Γ ◽

Peripheral Blood Mononuclear ◽

Colony Type ◽

Blood Mononuclear Cells ◽

Mycoplasma Mycoides ◽

Small Colony

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Monoclonal Antibody Differentiation ofMycoplasma mycoides subsp. mycoides Small-Colony Strains Causing Contagious Bovine Pleuropneumonia from Less Important Large-Colony Strains

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.7.3.519-521.2000 ◽

2000 ◽

Vol 7 (3) ◽

pp. 519-521 ◽

Cited By ~ 2

Author(s):

Fred R. Rurangirwa ◽

Patrick S. Shompole ◽

Anderson N. Wambugu ◽

Travis C. McGuire

Keyword(s):

Monoclonal Antibody ◽

Growth Inhibition ◽

In Vitro Growth ◽

Contagious Bovine Pleuropneumonia ◽

Large Colony ◽

Mycoplasma Mycoides ◽

Small Colony ◽

Polyclonal Antisera

ABSTRACT Monoclonal antibody (MAb) PK-2 inhibited the in vitro growth of nine Mycoplasma mycoides subsp. mycoidessmall-colony strains. In contrast to the results with polyclonal antisera, growth inhibition by MAb PK-2 was specific for M. mycoides subsp. mycoides small-colony strains and constituted a reliable means of distinguishing them from other mycoplasmas.

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Observations on the antigenic differences between the so-called SC and LC strains of Mycoplasma mycoides subsp. mycoides

Journal of Hygiene ◽

10.1017/s0022172400069680 ◽

1981 ◽

Vol 87 (3) ◽

pp. 437-442 ◽

Cited By ~ 6

Author(s):

G. R. Smith ◽

Janet C. Oliphant

Keyword(s):

Single Dose ◽

Protective Antigen ◽

Serological Tests ◽

Protective Antigens ◽

Large Colony ◽

Strain Vaccine ◽

Mycoplasma Mycoides ◽

Small Colony ◽

The Difference

SummaryThe so-called SC (small colony) and LC (large colony) strains of Mycoplasma mycoides subsp. mycoides are said to be indistinguishable by the in vitro serological tests generally used in mycoplasmology. In mice the immunity given by a single dose of killed LC-strain vaccine against challenge with SC strains is – unlike that given by SC-strain vaccine – only partial.When multiple doses of killed or living vaccines were given, the majority of 13 LC strains still failed to immunize completely against a SC strain. This suggests that, although some protective antigens are shared between both types of strain, at least one of importance is present in the SC strains but absent from the majority of LC strains. The difference between the protective-antigen content of SC and most LC strains is thus qualitative, and not merely quantitative.

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Demonstration of Mycoplasma capricolum subsp. capripneumoniae and Mycoplasma mycoides subsp. mycoides, Small Colony type in Outbreaks of Caprine Pleuropneumonia in Eastern Tanzania

Acta Veterinaria Scandinavica ◽

10.1186/bf03549639 ◽

2000 ◽

Vol 41 (3) ◽

pp. 311-319

Author(s):

L. J. M. Kusiluka ◽

W. D. Semuguruka ◽

R. R. Kazwala ◽

B. Ojeniy ◽

N. F. Friis

Keyword(s):

Colony Type ◽

Mycoplasma Capricolum ◽

Mycoplasma Mycoides ◽

Small Colony

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

Journal of Bacteriology ◽

10.1128/jb.184.13.3712-3722.2002 ◽

2002 ◽

Vol 184 (13) ◽

pp. 3712-3722 ◽

Cited By ~ 24

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.

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