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

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.

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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

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01582-05 ◽

2006 ◽

Vol 50 (8) ◽

pp. 2845-2849 ◽

Cited By ~ 10

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.

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Ulcerative balanitis and vulvitis of Dorper sheep in South Africa : a study on its aetiology and clinical features

Journal of the South African Veterinary Association ◽

10.4102/jsava.v76i4.426 ◽

2005 ◽

Vol 76 (4) ◽

Cited By ~ 8

Author(s):

A. Kidanemariam ◽

J. Gouws ◽

M. Van Vuuren ◽

B. Gummow

Keyword(s):

South Africa ◽

Clinical Features ◽

Clinical Signs ◽

Regular Feature ◽

Clinically Normal ◽

Large Colony ◽

Mycoplasma Mycoides ◽

Dorper Sheep ◽

Arcanobacterium Pyogenes ◽

Apparently Healthy

Ovine ulcerative balanitis and vulvitis in sheep of the Dorper breed has been observed in South Africa since 1979. Its aetiology has not been conclusively resolved, and there is some discrepancy in descriptions of its clinical features. In order to identify the pathogenic microorganism / s that contribute to the occurrence of the disease, the microflora in the genital tracts of both clinically healthy and affected sheep were isolated and compared. Bacteriological examination of materials from affected and unaffected sheep resulted in the isolation of Arcanobacterium pyogenes from 44.2 % and 17.2 % of them respectively. This difference is statistically significant (P < 0.01). Seventy-four per cent of the isolates originated from severe clinical cases. Mycoplasmas were isolated from 49.3 % of 116 clinically normal sheep and 78.2%of 104 affected sheep. There were significant differences in their rates of isolation in clinical groups (P < 0.05). Of all the mycoplasma isolates, Mycoplasma mycoides mycoides large colony variant (MmmLC) was isolated from 61.5 % of clinically diseased sheep while 6.0 % of the isolates were from apparently healthy animals (P < 0.05). The study threw light on the prevalence of mycoplasmas in the genital tract of apparently healthy sheep and, at the same time the identity of the mycoplasma pathogen associated with ulcerative balanitis and vulvitis was revealed. The findings of this investigation therefore confirmed the involvement of mycoplasma, particularly that of MmmLC large colony, in the disease in Dorper sheep in South Africa, and it was concluded that this microorganism is an important pathogen of balanitis and vulvitis in them. The study furthermore demonstrated a probable synergism between A. pyogenes and MmmLC. Finding these 2 organisms together occurred 53.4 times more frequently in the affected sheep than in the unaffected, which emphasises the probable multifactorial nature of the disease. The association between age and the presence of clinical signs was statistically significant. It was found that young sheep were more likely to have lesions than adult sheep. Clinical observations showed that the typical ulceration appears to be confined to the glans penis and lips of the vulva; no ulceration was observed on the shaft of the penis and prepuce or vaginal vestibule. In uncomplicated cases inflammation of the prepuce and vaginal vestibule is not a regular feature of the disease. Therefore the names ulcerative balanitis and vulvitis most accurately describe the nature of the disease in South Africa.

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Molecular characterisation of Mycoplasma species isolated from the genital tract of Dorper sheep in South Africa

Journal of the South African Veterinary Association ◽

10.4102/jsava.v86i1.1199 ◽

2015 ◽

Vol 86 (1) ◽

Cited By ~ 2

Author(s):

Habu A. Kalshingi ◽

Anna-Mari Bosman ◽

Johan Gouws ◽

Moritz Van Vuuren

Keyword(s):

South Africa ◽

Genital Tract ◽

The United States ◽

Mycoplasma Species ◽

Local Alignment ◽

Large Colony ◽

Mycoplasma Mycoides ◽

Search Tool ◽

Dorper Sheep ◽

Arcanobacterium Pyogenes

Biochemical and molecular analysis were conducted on 34 strains of Mycoplasma species isolated between 2003 and 2009 from the genital tract of clinically healthy Dorper sheep and sheep with ulcerative vulvitis and balanitis. Earlier publications identified the causative agent as Mycoplasma mycoides mycoides large colony (MmmLC) and Arcanobacterium pyogenes. The aims of the study were to characterise Mycoplasma species isolated from the genital tract of Dorper sheep with polymerase chain reaction assay, cloning and gene sequencing. Basic Local Alignment Search Tool (BLAST) results revealed six predominant Mycoplasma species: Mycoplasma arginini, Mycoplasma bovigenitalium, Arcanobacterium laidlawii, MmmLC, Mycoplasma sp. ovine/caprine serogroup II and M. canadense. Sequencing of the 34 isolates were analysed using phylogenetic methods, and 18 (50%) were identified as M. arginini with 99% – 100% similarity to M. arginini from England and Sweden. Six isolates showed 99% similarity to M. bovigenitalium strains from Turkey and Germany. Two isolates had 99% similarity to an M. sp. ovine/caprine sero group II from the United Kingdom. BLAST for two isolates revealed 99% similarity to Acholeplasma laidlawii from India, another two were 99% similar to MmmLC strain from Sweden, two showed 98% similarity to Mycoplasma sp. Usp 120 from Brazil, and two isolates have a 97% – 99% similarity to M. mm. Jcv1 strain from the United States of America. Finally, one isolate showed similarity of 99% to Mycoplasma canadense strain from Italy. The findings support the hypothesis that ulcerative vulvitis and balanitis of Dorper sheep in South Africa (SA) is a multifactorial disease with involvement of different Mycoplasma species.

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

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.64918-0 ◽

2007 ◽

Vol 57 (10) ◽

pp. 2247-2258 ◽

Cited By ~ 63

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.

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

Applied and Environmental Microbiology ◽

10.1128/aem.71.6.2888-2893.2005 ◽

2005 ◽

Vol 71 (6) ◽

pp. 2888-2893 ◽

Cited By ~ 30

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.

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