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Francois Thiaucourt - One of the best experts on this subject based on the ideXlab platform.
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Genome-wide analysis of the first sequenced Mycoplasma capricolum subsp capripneumoniae Strain M1601
G3, 2017Co-Authors: Shengli Chen, Ping Zhao, Yuefeng Chu, Francois Thiaucourt, Huafang Hao, Pengcheng Gao, Han Guo, Zhanhui Wang, Yongsheng LiuAbstract:Mycoplasma capricolum subsp. capripneumoniae (Mccp) is a common pathogen of goats that causes contagious caprine pleuropneumonia. We closed the gap and corrected rRNA operons in the draft genome of Mccp M1601: a strain isolated from an infected goat in a farm in Gansu, China. The genome size of M1601 is 1,016,707 bp with a GC content of 23.67%. We identified 915 genes (occupying 90.27% of the genome), of which 713 are protein-coding genes (excluding 163 pseudogenes). No genomic islands and complete insertion sequences were found in the genome. Putative determinants associated with the organism's virulence were analyzed, and 26 genes (including one adhesion protein gene, two capsule synthesis gene clusters, two lipoproteins, hemolysin A, ClpB, and proteins involved in pyruvate metabolism and cation transport) were potential virulence factors. In addition, two transporter systems (ATP-binding cassette [ABC] transporters and phosphotransferase) and two secretion systems (Sec and signal recognition particle [SRP] pathways) were observed in the Mccp genome. Genome synteny analysis reveals a good collinear relationship between M1601 and Mccp type strain F38. Phylogenetic analysis based on 11 single-copy core genes of 31 Mycoplasma strains revealed good collinearity between M1601 and Mycoplasma capricolum subsp. capricolum (Mcc) and close relationship among Mycoplasma mycoides cluster strains. Our genome-wide analysis of Mccp M1601 provides helpful information on the pathogenic mechanisms and genetics of Mccp.
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complete genome sequence of Mycoplasma capricolum subsp capripneumoniae strain 9231 abomsa
Genome Announcements, 2014Co-Authors: Virginie Dupuy, Francois ThiaucourtAbstract:Mycoplasma capricolum subsp. capripneumoniae is the etiological agent of contagious caprine pleuropneumonia. We report here the complete and annotated genome sequence of M. capricolum subsp. capripneumoniae strain 9231-Abomsa. (Resume d'auteur)
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fatal outbreak of Mycoplasma capricolum pneumonia in endangered markhors
Emerging Infectious Diseases, 2011Co-Authors: Stephane Ostrowski, Virginie Dupuy, Francois Thiaucourt, Mulojon Amirbekov, Abdurahmon Mahmadshoev, Lucia Mansosilvan, Dustmurod Vahobov, Orom Ziyoev, Stefan MichelAbstract:Mycoplasma capricolum subsp. capricolum and M. capricolum subsp. capripneumoniae are closely related subspecies of the M. mycoides cluster (1). Whereas M. capricolum subsp. capripneumoniae is the etiologic agent of contagious caprine pleuropneumonia (CCPP), a severe and typically lethal respiratory disease, M. capricolum subsp. capricolum infection is usually not fatal and instead results in chronic inflammation in a variety of organs, including joints, udder, eyes, and lungs (2). M. capricolum subsp. capricolum infection occurs worldwide and appears widespread but has rarely been found in species of small ruminants other than domestic goats and, more occasionally, sheep (2,3). This lack of evidence may be partially because few studies have applied sensitive molecular techniques for its detection in nondomestic ruminants (2,3). Domestic goats can carry M. capricolum subsp. capricolum asymptomatically, notably in the ear canal (4), and pose an insidious risk for cross-species transmission with sympatric wild caprines (2,3).
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multi locus sequence analysis of Mycoplasma capricolum subsp capripneumoniae for the molecular epidemiology of contagious caprine pleuropneumonia
Veterinary Research, 2011Co-Authors: Lucia Mansosilvan, Virginie Dupuy, Francois ThiaucourtAbstract:Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the causative agent of contagious caprine pleuropneumonia (CCPP), a devastating disease of domestic goats. The exact distribution of CCPP is not known but it is present in Africa and the Middle East and represents a significant threat to many disease-free areas including Europe. Furthermore, CCPP has been recently identified in Tajikistan and China. A typing method with an improved resolution based on Multi-Locus Sequence Analysis (MLSA) has been developed to trace new epidemics and to elucidate whether the recently identified cases in continental Asia were due to recent importation of Mccp. The H2 locus, a polymorphic region already in use as a molecular marker for Mccp evolution, was complemented with seven new loci selected according to the analysis of polymorphisms observed among the genome sequences of three Mccp strains. A total of 25 strains, including the two new strains from Asia, were analysed by MLSA resulting in the discrimination of 15 sequence types based on 53 polymorphic positions. A distance tree inferred from the concatenated sequences of the eight selected loci revealed two evolutionary lineages comprising five groups, which showed good correlation with geographic origins. The presence of a distinct Asian cluster strongly indicates that CCPP was not recently imported to continental Asia. It is more likely that the disease has been endemic in the area for a long time, as supported by historical clinical descriptions. In conclusion, this MLSA strategy constitutes a highly discriminative tool for the molecular epidemiology of CCPP.
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specific real time pcr assays for the detection and quantification of Mycoplasma mycoides subsp mycoides sc and Mycoplasma capricolum subsp capripneumoniae
Molecular and Cellular Probes, 2008Co-Authors: Sophie Lorenzon, Lucia Mansosilvan, Francois ThiaucourtAbstract:Abstract Contagious bovine pleuropneumonia and contagious caprine pleuropneumonia are two severe respiratory infections of ruminants due to infection by Mycoplasma mycoides subsp. mycoides SC (MmmSC) and Mycoplasma capricolum subsp. capripneumoniae (Mccp), respectively. They are included in the OIE list of notifiable diseases. Here we describe the development of rapid, sensitive, and specific real-time PCR assays for the detection and quantification of MmmSC and Mccp DNA. MmmSC PCR primers were designed after whole genome comparisons between the published sequence of MmmSC strain type PG1T and the sequence of an M. mycoides subsp. mycoides large colony strain. For Mccp, previously published conventional PCR primers were applied. SYBR green was used as a detection agent for both assays. The assays specifically detected the targeted species in both cultures and clinical samples, and no cross-amplifications were obtained from either heterologous Mycoplasma strain cultures or European field samples. The sensitivity of these new assays was estimated at 3–80 colony forming units per reaction and 4–80 fg of DNA, representing a 2–3 log increase in sensitivity compared to established conventional PCR tests. These new real-time PCR assays will be invaluable for application in various fields such as direct detection in diagnostic laboratories.
Xiaobo Duan - One of the best experts on this subject based on the ideXlab platform.
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genome sequence of Mycoplasma capricolum subsp capripneumoniae strain m1601
Journal of Bacteriology, 2011Co-Authors: Yuefeng Chu, Ping Zhao, Pengchen Gao, Nancy Y Liao, Shaun D Jackman, Yongjun Zhao, Inanc Birol, Xiaobo DuanAbstract:ABSTRACT Mycoplasma capricolum subsp. capripneumoniae is the causative agent of contagious caprine pleuropneumonia, a devastating disease of goats listed by the World Organization for Animal Health. Here we report the first complete genome sequence of this organism (strain M1601, a clinically isolated strain from China).
Karlerik Johansson - One of the best experts on this subject based on the ideXlab platform.
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genetic diversity and evolution of Mycoplasma capricolum subsp capripneumoniae strains from eastern africa assessed by 16s rdna sequence analysis
Veterinary Microbiology, 2001Co-Authors: Malin U K Heldtander, Goran Bolske, Bertil Pettersson, Hezron Wesonga, Karlerik JohanssonAbstract:Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae), the causal agent of contagious caprine pleuropneumonia (CCPP), is a member of the so-called Mycoplasma mycoides cluster. These Mycoplasmas have two rRNA operons in which intraspecific variations have been demonstrated. The sequences of the 16S rRNA genes of both operons from 13 field strains of M. capripneumoniae from three neighbouring African countries (Kenya, Ethiopia, and Tanzania) were determined. Four new and unique polymorphism patterns reflecting the intraspecific variations were found. Two of these patterns included length differences between the rrnA and rrnB operons. The length difference in one of the patterns was caused by a two-nucleotide insert (TG) in the rrnB operon and the length difference in the other pattern was due to a three-nucleotide deletion, also in the rrnB operon. Another pattern was characterised by a polymorphic position caused by a mutation that is known to cause streptomycin resistance in other bacterial species. The strain with this pattern was also found to be resistant to streptomycin. Streptomycin resistant clones were selected from four M. capripneumoniae strains to further investigate the correlation of this mutation to streptomycin resistance. Mutations in the 16S rRNA genes had occurred in two of these strains. The fourth pattern included a new polymorphism in position 1059. The results show that polymorphisms in M. capripneumoniae strains can be used as epidemiological markers for CCPP in smaller geographical areas and to study the molecular evolution of this species.
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genomic variations of Mycoplasma capricolum subsp capripneumoniae detected by amplified fragment length polymorphism aflp analysis
Fems Microbiology Letters, 2000Co-Authors: Branko Kokotovic, Goran Bolske, Peter Ahrens, Karlerik JohanssonAbstract:The genetic diversity of Mycoplasma capricolum subsp. capripneumoniae strains based on determination of amplified fragment length polymorphisms (AFLP) is described. AFLP fingerprints of 38 strains derived from different countries in Africa and the Middle East consisted of over 100 bands in the size range of 40-500 bp. The similarity between individual AFLP profiles, calculated by Jaccard's coefficient, ranged from 0.92 to 1.0. On the basis of the polymorphisms detected, the analysed strains can explicitly be grouped into two major clusters, equivalent to two evolutionary lines of the organism found by 16S rDNA analysis. The present data support previous observations regarding genetic homogeneity of M. capricolum subsp. capripneumoniae, and confirm the two evolutionary lines of descent found by analysis of 16S rRNA genes.
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molecular evolution of Mycoplasma capricolum subsp capripneumoniae strains based on polymorphisms in the 16s rrna genes
Journal of Bacteriology, 1998Co-Authors: Bertil Pettersson, Francois Thiaucourt, Goran Bolske, Karlerik Johansson, Mathias UhlenAbstract:Mycoplasma capricolum subsp. capripneumoniae belongs to the so-called Mycoplasma mycoides cluster and is the causal agent of contagious caprine pleuropneumonia (CCPP). All members of the M. mycoides cluster have two rRNA operons. The sequences of the 16S rRNA genes of both rRNA operons from 20 strains of M. capricolum subsp. capripneumoniae of different geographical origins in Africa and Asia were determined. Nucleotide differences which were present in only one of the two operons (polymorphisms) were detected in 24 positions. The polymorphisms were not randomly distributed in the 16S rRNA genes, and some of them were found in regions of low evolutionary variability. Interestingly, 11 polymorphisms were found in all the M. capricolum subsp. capripneumoniae strains, thus defining a putative ancestor. A sequence length difference between the 16S rRNA genes in a poly(A) region and 12 additional polymorphisms were found in only one or some of the strains. A phylogenetic tree was constructed by comparative analysis of the polymorphisms, and this tree revealed two distinct lines of descent. The nucleotide substitution rate of strains within line II was up to 50% higher than within line I. A tree was also constructed from individual operonal 16S rRNA sequences, and the sequences of the two operons were found to form two distinct clades. The topologies of both clades were strikingly similar, which supports the use of 16S rRNA sequence data from homologous operons for phylogenetic studies. The strain-specific polymorphism patterns of the 16S rRNA genes of M. capricolum subsp. capripneumoniae may be used as epidemiological markers for CCPP.
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evolution moleculaire des types de Mycoplasma capricolum subsp capripneumoniae basee sur les polymorphismes dans les genes rrna 16s
Journal of Bacteriology, 1998Co-Authors: Bertil Pettersson, Francois Thiaucourt, Goran Bolske, Mathias Uhlen, Karlerik JohanssonAbstract:Mycoplasma capricolum subsp. capripneumoniae belongs to the so-called Mycoplasma mycoides cluster and is the causal agent of contagious caprine pleuropneumonia (CCPP). All members of the M. mycoides cluster have two rRNA operons. The sequences of the 16S rRNA genes of both rRNA operons from 20 strains of M. capricolum subsp. capripneumoniae of different geographical origins in Africa and Asia were determined. Nucleotide differences which were present in only one of the two operons (polymorphisms) were detected in 24 positions. The polymorphisms were not randomly distributed in the 16S rRNA genes, and some of them were found in regions of low evolutionary variability. Interestingly, 11 polymorphisms were found in all the M. capricolum subsp. capripneumoniae strains, thus defining a putative ancestor. A sequence length difference between the 16S rRNA genes in a poly(A) region and 12 additional polymorphisms were found in only one or some of the strains. A phylogenetic tree was constructed by comparative analysis of the polymorphisme, and this tree revealed two distinct fines of descent. The nucleotide substitution rate of strains within line II was up to 50% higher than within line I. A tree was also constructed from individual operonal 16S rRNA sequences, and the sequences of the two operons were found to form two distinct clades. The topologies of both clades were strikingly similar, which supports the use of 16S rRNA sequence data from homologous operons for phylogenetic studies. The strain-specific polymorphism patterns of the 16S rRNA genes of M. capricolum subsp. capripneumoniae may be used as epidemiological markers for CCPP. (Resume d'auteur)
Joachim Frey - One of the best experts on this subject based on the ideXlab platform.
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versatile use of oric plasmids for functional genomics of Mycoplasma capricolum subsp capricolum
Applied and Environmental Microbiology, 2005Co-Authors: Carole Janis, Francois Thiaucourt, Joachim Frey, Carole Lartigue, Henri Wroblewski, Alain Blanchard, Pascal SirandpugnetAbstract: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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comparative analysis of the lppa locus in Mycoplasma capricolum subsp capricolum and Mycoplasma capricolum subsp capripneumoniae
Veterinary Microbiology, 1999Co-Authors: Mariepierre Monnerat, Francois Thiaucourt, J Nicolet, Joachim FreyAbstract:The lppA gene, encoding the lipoprotein named LppA[Mcaca] was characterised in Mycoplasma capricolum subsp. capricolum. It encodes a lipoprotein with an apparent molecular mass of 57 kDa as determined by SDS-PAGE. Using antibodies directed against recombinant LppA[Mcaca], we showed the expression of this lipoprotein in all M. capricolum subsp. capricolum by immunoblot analysis. The serum did not cross-react with other members of the Mycoplasma mycoides cluster, hence showing that LppA[Mcaca] was antigenically specific to M. capricolum subsp. capricolum. The lppA gene was conserved within the subspecies and was used for the development of a specific PCR assay for the identification of M. capricolum subsp. capricolum. The taxonomically related Mycoplasma capricolum subsp. capripneumoniae (F38) was found to contain an lppA-pseudo-gene. It showed high similarity to functional lppA genes of other Mycoplasmas in the M. mycoides cluster. However, it contained interrupted open reading frames. Moreover, the nucleotide sequence of the lppA pseudo-genes in different strains of M. capricolum subsp. capripneumoniae were quite variable. Interestingly, the lppA pseudo-gene had a size similar to that of the functional lppA genes of other Mycoplasmas of the M. mycoides cluster and occupied the same genomic location as the latter ones in the vicinity of the mtlD genes. This study showed that all members of the M. mycoides cluster contain each a species-, subspecies- respectively type- specific lppA gene analogue which encodes a lipoprotein that has structural and functional relationship to the surface lipoprotein LppA [MmymySC], previously named P72, of M. mycoides subsp mycoides SC, with the exception of M. capricolum subsp. capripneumoniae which seems not to express an LppA analogue.
Lucia Mansosilvan - One of the best experts on this subject based on the ideXlab platform.
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fatal outbreak of Mycoplasma capricolum pneumonia in endangered markhors
Emerging Infectious Diseases, 2011Co-Authors: Stephane Ostrowski, Virginie Dupuy, Francois Thiaucourt, Mulojon Amirbekov, Abdurahmon Mahmadshoev, Lucia Mansosilvan, Dustmurod Vahobov, Orom Ziyoev, Stefan MichelAbstract:Mycoplasma capricolum subsp. capricolum and M. capricolum subsp. capripneumoniae are closely related subspecies of the M. mycoides cluster (1). Whereas M. capricolum subsp. capripneumoniae is the etiologic agent of contagious caprine pleuropneumonia (CCPP), a severe and typically lethal respiratory disease, M. capricolum subsp. capricolum infection is usually not fatal and instead results in chronic inflammation in a variety of organs, including joints, udder, eyes, and lungs (2). M. capricolum subsp. capricolum infection occurs worldwide and appears widespread but has rarely been found in species of small ruminants other than domestic goats and, more occasionally, sheep (2,3). This lack of evidence may be partially because few studies have applied sensitive molecular techniques for its detection in nondomestic ruminants (2,3). Domestic goats can carry M. capricolum subsp. capricolum asymptomatically, notably in the ear canal (4), and pose an insidious risk for cross-species transmission with sympatric wild caprines (2,3).
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multi locus sequence analysis of Mycoplasma capricolum subsp capripneumoniae for the molecular epidemiology of contagious caprine pleuropneumonia
Veterinary Research, 2011Co-Authors: Lucia Mansosilvan, Virginie Dupuy, Francois ThiaucourtAbstract:Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the causative agent of contagious caprine pleuropneumonia (CCPP), a devastating disease of domestic goats. The exact distribution of CCPP is not known but it is present in Africa and the Middle East and represents a significant threat to many disease-free areas including Europe. Furthermore, CCPP has been recently identified in Tajikistan and China. A typing method with an improved resolution based on Multi-Locus Sequence Analysis (MLSA) has been developed to trace new epidemics and to elucidate whether the recently identified cases in continental Asia were due to recent importation of Mccp. The H2 locus, a polymorphic region already in use as a molecular marker for Mccp evolution, was complemented with seven new loci selected according to the analysis of polymorphisms observed among the genome sequences of three Mccp strains. A total of 25 strains, including the two new strains from Asia, were analysed by MLSA resulting in the discrimination of 15 sequence types based on 53 polymorphic positions. A distance tree inferred from the concatenated sequences of the eight selected loci revealed two evolutionary lineages comprising five groups, which showed good correlation with geographic origins. The presence of a distinct Asian cluster strongly indicates that CCPP was not recently imported to continental Asia. It is more likely that the disease has been endemic in the area for a long time, as supported by historical clinical descriptions. In conclusion, this MLSA strategy constitutes a highly discriminative tool for the molecular epidemiology of CCPP.
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specific real time pcr assays for the detection and quantification of Mycoplasma mycoides subsp mycoides sc and Mycoplasma capricolum subsp capripneumoniae
Molecular and Cellular Probes, 2008Co-Authors: Sophie Lorenzon, Lucia Mansosilvan, Francois ThiaucourtAbstract:Abstract Contagious bovine pleuropneumonia and contagious caprine pleuropneumonia are two severe respiratory infections of ruminants due to infection by Mycoplasma mycoides subsp. mycoides SC (MmmSC) and Mycoplasma capricolum subsp. capripneumoniae (Mccp), respectively. They are included in the OIE list of notifiable diseases. Here we describe the development of rapid, sensitive, and specific real-time PCR assays for the detection and quantification of MmmSC and Mccp DNA. MmmSC PCR primers were designed after whole genome comparisons between the published sequence of MmmSC strain type PG1T and the sequence of an M. mycoides subsp. mycoides large colony strain. For Mccp, previously published conventional PCR primers were applied. SYBR green was used as a detection agent for both assays. The assays specifically detected the targeted species in both cultures and clinical samples, and no cross-amplifications were obtained from either heterologous Mycoplasma strain cultures or European field samples. The sensitivity of these new assays was estimated at 3–80 colony forming units per reaction and 4–80 fg of DNA, representing a 2–3 log increase in sensitivity compared to established conventional PCR tests. These new real-time PCR assays will be invaluable for application in various fields such as direct detection in diagnostic laboratories.
