Application of dot immunobinding on membrane filtration (MF dot) to the study of relationships within “M. mycoides cluster” and within “glucose and arginine-negative cluster” of ruminant mycoplasmas
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High-Throughput Sequencing (HTS) of newly synthetized RNAs enables one shot detection and identification of live mycoplasmas and differentiation from inert nucleic acids
2020, BiologicalsCitation Excerpt :Dilution series of a 107 CFU/mL stock culture of A. laidlawii strain PG8T (also named ATCC23206, NCTC10116, which has been maintained in the ANSES facilities for years) were used for infection assays. The number of CFUs per ml was confirmed by plating 2 μL of serial 10-fold broth dilutions onto pleuropneumonia-like organisms (PPLO) agar plates (Indicia, France), modified as previously described [13]. After incubation for 3 days at 37 °C in 5% CO2, the colonies of several dilutions were counted using a stereomicroscope and the mean final concentration of 107 CFU/mL was confirmed.
Loss of diversity within Mycoplasma bovis isolates collected in France from bovines with respiratory diseases over the last 35years
2015, Infection, Genetics and EvolutionCitation Excerpt :These were the type strain PG45 (Wise et al., 2011) with accession number NC_014760 as well as two isolates from China, namely Hubei-1 (Li et al., 2011) and HB0801 (Qi et al., 2012), with accession numbers NC_015725 and NC_018077, respectively. Strains were grown for 48 h at 37 °C under 5% CO2, in PPLO broth (Difco), supplemented as previously described (Poumarat et al., 1992). Genomic DNA was extracted using manufactured kits (Blood & Cell Culture DNA kit, Qiagen).
Development of a multiplex real-time PCR for contagious agalactia diagnosis in small ruminants
2012, Journal of Microbiological MethodsCitation Excerpt :These comprised a wide variety of CA-isolates (varied locations, clinical expressions and host origins) and several non CA-associated mycoplasmas. Strains were grown at 37 °C under 5% CO2 in PPLO agar/broth (Difco, Le Pont de Claix, France), supplemented as previously described (Poumarat et al., 1992). Genomic DNAs were extracted from 1 ml of mycoplasmal culture in stationary phase using the DNeasy tissue kit (Qiagen, Courtaboeuf, France).
Extended surveillance for CBPP in a free country: Challenges and solutions regarding the potential caprine reservoir
2011, Preventive Veterinary MedicineCitation Excerpt :Several isolates were included in the present study, either collected in the context of the French surveillance network for mycoplasmoses (Chazel et al., 2010) or obtained from other collections (list in Supplementary information). They were grown in PPLO agar/broth (Difco, France), supplemented as previously described (Poumarat et al., 1992), at 37 °C under 5% CO2 and were identified using membrane-filtration immunobinding tests (MF-dot), the current methodology for routine strain identification in the laboratory (Tardy et al., 2009a). The fusA gene, shown to be useful for taxon identification within the M. mycoides cluster (Manso-Silván et al., 2007; Maigre et al., 2008), was also sequenced.
Comparison of Isolates of Mycoplasma mycoides subspecies capri from Asymptomatic and Septicaemic Goats
2011, Journal of Comparative PathologyCitation Excerpt :The carriage strains (ear canal [EC] or BTM) were isolated over the same period from five different herds in the Poitou-Charentes region. Mycoplasma strains were grown at 37°C under 5% CO2 in PPLO broth (Difco; Le Pont de Claix, France) supplemented as detailed previously (Poumarat et al., 1992). When necessary, the growth of these isolates was assessed by cell counting using a Most Probable Number method (Cochran, 1950).