A specific PCR for the detection of Mycoplasma putrefaciens, one of the agents of the contagious agalactia syndrome of goats

Peyraud, Armelle; Woubit, Salah; Fe, Christian de la; Mercier, Pascale; Thiaucourt, François

doi:10.1016/j.mcp.2003.07.006

Cited by 36 publications

(8 citation statements)

References 18 publications

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“…agalactiae was conducted by real time PCR as previously described [11] in a 7500 real time PCR system platform (Applied Bios stems, Carlsbad, CA, USA). M. mycoides cluster and M. putrefaciens detection was carried out by conventional PCR [12,13]. PCR amplification products were resolved by gel electrophoresis on 1.5% (w/v) agarose gels and visualized after staining with ethidium bromide by using a UV transilluminator (BioRad, Hercules, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

A survey of Mycoplasma agalactiaein dairy sheep farms in Spain

2012

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BackgroundContagious Agalactia (CA) is one of the major animal health problems in small ruminants because of its economic significance. Currently, four Mycoplasma spp. have been associated with this syndrome: M. agalactiae, M. mycoides subsp. capri, M. capricolum subsp. capricolum and M. putrefaciens. Their presence has been evaluated in several studies conducted in CA-endemic countries. However, previous Spanish studies have been focused on caprine CA, and there is a knowledge gap regarding which Mycoplasma species are present in sheep flocks from Spain, which has the second highest number of sheep amongst the 27 European Union member states. Consequently, we investigated the presence and geographic distribution of the four CA-causing mycoplasmas in Spanish dairy sheep farms. This is the first time such an investigation has been performed.ResultsThree hundred thirty nine out of 922 sheep flocks were positive for M. agalactiae by real time PCR (36.8%) and 85 by microbiological identification (9.2%). Interestingly, all 597 milk samples assessed for the presence of M. mycoides subsp. capri, M. capricolum subsp. capricolum and M. putrefaciens tested negative. To evaluate the intermittent excretion of the pathogen in milk, we sampled 391 additional farms from 2 to 5 times, resulting that in 26.3% of the cases a previously positive farm tested negative in a later sampling.ConclusionsM. agalactiae was the only Mycoplasma species detected in the study area showing a high frequency of presence and wide distribution. Therefore, the establishment of a permanent surveillance network is advantageous, as well as the implementation of control and prevention measures to hinder the dissemination of M. agalactiae and to prevent the entrance of other Mycoplasma species.

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Section: Methodsmentioning

confidence: 99%

A survey of Mycoplasma agalactiaein dairy sheep farms in Spain

2012

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“…The PCR procedures used were specific for Ma and Mp [33,34]. To detect the presence of Mmc or M capricolum subsp capricolum (Mcc), an initial PCR was conducted to detect all members of the M mycoides cluster [35].…”

Section: Mycoplasma Culture and Pcrmentioning

confidence: 99%

Survival capacity of Mycoplasma agalactiae and Mycoplasma mycoides subsp capri in the diluted semen of goat bucks and their effects on sperm quality

Gómez-Martín

Vieira

et al. 2015

Theriogenology

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“…The resulting, typical "fried-egg" Mycoplasma colonies were examined by light microscopy at 10ϫ magnification. In order to confirm that the isolated colonies were M. agalactiae, we tested the enrichment broth cultures for the presence of strains of Mycoplasma species involved in contagious agalactia in small ruminants (i.e., M. mycoides cluster and M. putrefaciens), using PCR methods previously published (18,29).…”

Section: Capri M Mycoides Subsp Mycoides Lc and M Putrefaciens)mentioning

confidence: 99%

Mycoplasma agalactiae p40 Gene, a Novel Marker for Diagnosis of Contagious Agalactia in Sheep by Real-Time PCR: Assessment of Analytical Performance and In-House Validation Using Naturally Contaminated Milk Samples

et al. 2009

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We evaluated the capacity of the Mycoplasma agalactiae p40 gene as a diagnostic marker for contagious agalactia in sheep by quantitative real-time PCR. The p40 gene encodes an immunodominant adhesin that plays a key role in cytoadhesion of M. agalactiae. The assay was 100% specific, with an analytical sensitivity of 1 genome equivalent (GE), a quantification that is highly linear (R 2 > 0.992) and efficient (PCR efficiency, >0.992) over a 6-log dynamic range, down to 10 GE. We evaluated the capacity of the assay to detect Mycoplasma agalactiae in 797 milk samples (373 raw sheep milk samples from refrigerated tanks of different farms and 424 milk samples from individual sheep of a flock positive for M. agalactiae). In parallel, we also tested the samples by using microbiological isolation coupled with microscopy identification and by a PCR method recommended by the World Organization for Animal Health. While our assay was able to detect 57 (15.28%) positive samples of the 373 milk samples from different farms, identification by microbiological isolation coupled with microscopy detected only 36 (9.65%) samples, and the conventional PCR detected 31 (8.31%) samples. These findings showed that our assay based on the p40 gene is more specific and sensitive for the detection of M. agalactiae in actual natural samples and, thus, can be a promising alternative tool for diagnosis and epidemiological studies of M. agalactiae infection.

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A specific PCR for the detection of Mycoplasma putrefaciens, one of the agents of the contagious agalactia syndrome of goats

Cited by 36 publications

References 18 publications

A survey of Mycoplasma agalactiaein dairy sheep farms in Spain

A survey of Mycoplasma agalactiaein dairy sheep farms in Spain

Survival capacity of Mycoplasma agalactiae and Mycoplasma mycoides subsp capri in the diluted semen of goat bucks and their effects on sperm quality

Mycoplasma agalactiae p40 Gene, a Novel Marker for Diagnosis of Contagious Agalactia in Sheep by Real-Time PCR: Assessment of Analytical Performance and In-House Validation Using Naturally Contaminated Milk Samples

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