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 Use of flow cytometry for enumeration of Mycoplasma mycoides subsp. mycoides large-colony type in broth medium

2006 ◽  
Vol 100 (4) ◽  
pp. 878-884 ◽  
Author(s):  
P. Assuncao ◽  
C. Fe ◽  
N.T. Antunes ◽  
R.S. Rosales ◽  
C.M. Ruiz de Galarreta ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Colony Type ◽  
Large Colony ◽  
Mycoplasma Mycoides

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 Chromosome doubling in Cattleya tigrina A. Rich

2019 ◽  
Vol 15 (11) ◽  
Author(s):  
Thays Saynara Alves Menezes-Sá ◽  
Maria de Fátima Arrigoni-Blank ◽  
Andréa Santos da Costa ◽  
Janay De Almeida Santos-Serejo ◽  
Arie Fitzgerald Blank ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Propidium Iodide ◽  
Chromosome Doubling ◽  
Flow Cytometric Analysis ◽  
Leaf Tissue ◽  
Stomatal Index ◽  
Chromosome Duplication ◽  
Flow Cytometric ◽  
Young Leaves ◽  
Commercial Value

Chromosome doubling induction in orchids may benefit their production for resulting in flowers of higher commercial value, larger size and higher content of substances that intensify the color and fragrance when compared with diploid orchids. This work aimed to induce and confirm artificial polyploidization, using flow cytometry and stomatal analysis. Explants were treated with colchicine at concentrations of 0, 2.5, 7.5, and 12.5 mM, for 24 and 48 hours and with oryzalin, at concentrations of 0, 10, 30, and 50 μM, for three and six days. For the flow cytometric analysis, a sample of leaf tissue was removed from each plant, crushed to release the nuclei and stained with propidium iodide. In addition to flow cytometry, the ploidy of the antimitotic treated plants was evaluated by stomata analysis. Young leaves were used where the density, functionality and stomatal index were evaluated. Colchicine provided induction of satisfactory polyploidy in C. tigrina at all concentrations and times of exposure, obtaining a greater number of polyploid individuals in the concentration of 12.5 mM for 48 hours. Oryzalin did not induce chromosome duplication at the tested concentrations.

scholarly journals Flow cytometric analysis of herpes simplex virus type 1 susceptibility to acyclovir, ganciclovir, and foscarnet.

1997 ◽  
Vol 41 (12) ◽  
pp. 2686-2692 ◽  
Author(s):  
I Pavić ◽  
A Hartmann ◽  
A Zimmermann ◽  
D Michel ◽  
W Hampl ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Cytopathic Effect ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Resistant Strains ◽  
Simplex Virus

We established a quantitative flow cytometric method for determination of herpes simplex virus type 1 (HSV-1) susceptibility to acyclovir (ACV), ganciclovir, and foscarnet in vitro. Susceptibility was defined in terms of the drug concentration which reduced the number of cells expressing HSV-1 glycoprotein C (gpC) with a fluorescence intensity of > or =10(2) by 50% (IC50). Flow cytometry allowed us to use a high (1.0) as well as a low (0.005) multiplicity of infection, and determination of the IC50 was possible after one or more viral replicative cycles. IC50s were dependent on virus input and on time postinfection. In mixture experiments, 1 to 2% resistant viruses added to a sensitive strain could be detected. The results obtained by flow cytometry showed a good qualitative correlation with those achieved by cytopathic effect inhibitory assay. However, flow cytometry might detect more quantitative differences in drug susceptibility, especially among resistant strains, as confirmed also by determination of intracellular drug phosphorylation. The mean IC50s for ACV-sensitive strains were 0.45 to 1.47 microM, and those for ACV-resistant strains were between 140 and 3,134 microM. Flow cytometric analysis was fast and accurate, automatizable, and highly reproducible. Flow cytometry may be a more powerful tool than standard cytopathic effect-based assays and could have advantages for the detection of low levels of drug resistance or mixtures of sensitive and resistant virus strains.

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

2015 ◽  
Vol 86 (1) ◽  
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.

Sign in / Sign up

Export Citation Format

Share Document