A new microtitration method for the enumeration of contagious bovine pleuropneumonia (CBPP) vaccines

Thiaucourt, François; Maria, Annalisa Di

doi:10.1016/s1045-1056(05)80002-8

Cited by 10 publications

(9 citation statements)

References 6 publications

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“…Mycoplasma medium or simply pleuropneumonia-like organism (PPLO) basal medium (Difco Laboratory, Detroit, USA) was prepared according to the standard method (Thiaucourt & Di-Maria, 1992). This medium is effective for in vitro antimicrobial susceptibility testing (Hannan, 2000).…”

Section: Antimycoplasmal Assaysmentioning

confidence: 99%

Antimycoplasmal activity of some plant species from northern Nigeria compared to the currently used therapeutic agent

Muraina¹,

Adaudi²,

Mamman³

et al. 2010

Pharmaceutical Biology

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Section: Antimycoplasmal Assaysmentioning

confidence: 99%

Antimycoplasmal activity of some plant species from northern Nigeria compared to the currently used therapeutic agent

Muraina¹,

Adaudi²,

Mamman³

et al. 2010

Pharmaceutical Biology

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“…Strains were grown in a modified Hayflick medium containing sodium pyruvate and glucose [39]. At the end of the exponential phase of growth, 2 mL of culture were centrifuged at 12 000 × g for 15 min at 4 • C. The pellet was washed once in PBS, and re-suspended in 100 µL of distilled water, to which 150 µL of lysis buffer (100 mM Tris HCl, pH 8.5; 0.05% Tween 20; 0.24 mg/mL proteinase K) were added.…”

Section: Mmmsc Strains and Isolatesmentioning

confidence: 99%

Genotyping ofMycoplasma mycoidessubsp.mycoidesSC by multilocus sequence analysis allows molecular epidemiology of contagious bovine pleuropneumonia

et al. 2008

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-Mycoplasma mycoides subsp. mycoides SC (MmmSC) is the etiological agent of contagious bovine pleuropneumonia (CBPP). Although eradicated in most developed countries, the disease reappeared in Europe in the 1990s. This reappearance may have been caused either by importation from sub-Saharan Africa, where CBPP is still endemic, or by the reemergence of virulent strains in Europe, as suggested by earlier studies. A multilocus sequence analysis scheme has been developed to address this issue and, most importantly, to be able to monitor new epidemics. The alignment of the full genome sequence of the reference strain PG1 and the partial genome sequence of a pathogenic strain allowed the identification of polymorphic sites. Nineteen initial loci were selected within housekeeping genes, genes of unknown function and non coding sequences. The suitability of these loci for genotyping MmmSC strains was first tested on six strains of diverse geographic origin. The analyses showed that the published PG1 sequence contained a number of specific polymorphisms that were therefore of no use for molecular typing. Among the eight informative polymorphic loci finally selected, only one (ftsY) was positioned within a housekeeping gene. Three main groups and 31 different allelic profiles were identified among 51 strains and strain variants examined. Cluster analysis confirmed that European strains from the 1990s did not originate from Africa. It also showed a genetic link between a European strain isolated in 1967 and those found in southern Africa and Australia. This was in agreement with historical data showing that CBPP was introduced in these regions during colonisation in the 19th century.Mycoplasma mycoides / contagious bovine pleuropneumonia / multilocus sequence analysis / molecular epidemiology

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“…The pH of 1-ml samples was measured using a pH meter (Mettler, Delta 345). The titre (colour changing units (CCU) per ml) was determined using a microtitration method adapted from [22]. Brie£y, 50 Wl of the growing culture was added to the ¢rst column of a sterile 96-well microtitre plate.…”

Section: Determination Of the Ph And Titrementioning

confidence: 99%

Effect of HEPES buffer systems upon the pH, growth and survival of Mycoplasma mycoides subsp. mycoides small colony (MmmSC) vaccine cultures

Waite¹

2001

FEMS Microbiology Letters

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The use of a buffer system based on N-[2-hydroxyethyl]piperazine-NP-[2-ethanesulfonic acid] (HEPES), in conjunction with standard Gourlay's culture medium was investigated for the growth and maintenance of Mycoplasma mycoides subsp. mycoides SC vaccine strain T 1 44. When the initial pH of the culture medium was adjusted to 8.0, 0.075 M HEPES^NaOH was found to be sufficient to prevent the pH falling below 7.1 at any stage during the growth cycle, even in the presence of 0.5% glucose. Compared to growth in standard unbuffered Gourlay's medium, the final culture titre was found to be one log 10 higher, at 10 11 colour changing units (CCU) per ml, and considerably extended culture survival was observed at 37³C. The titre remained above 10 10 CCU ml 31 for 4 days, and above 10 8 CCU ml 31 in excess of 1 month. After 4 month's storage at 37³C the titre had fallen to 5U10 4 CCU ml 31 . In contrast, no viable bacteria could be detected in standard unbuffered medium 3 days after the onset of stationary phase, at which point the pH had dropped to 5.4. No significant difference in growth rate between the two media was observed. Adoption of a HEPES^NaOH buffer system by African vaccine manufacturers should require minimal changes to current formulations and procedures, and should enhance both the final titre and thermostability of freeze-dried and liquid broth vaccines against contagious bovine pleuropneumonia (CBPP). ß

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A new microtitration method for the enumeration of contagious bovine pleuropneumonia (CBPP) vaccines

Cited by 10 publications

References 6 publications

Antimycoplasmal activity of some plant species from northern Nigeria compared to the currently used therapeutic agent

Antimycoplasmal activity of some plant species from northern Nigeria compared to the currently used therapeutic agent

Genotyping ofMycoplasma mycoidessubsp.mycoidesSC by multilocus sequence analysis allows molecular epidemiology of contagious bovine pleuropneumonia

Effect of HEPES buffer systems upon the pH, growth and survival of Mycoplasma mycoides subsp. mycoides small colony (MmmSC) vaccine cultures

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