Publications (8)19.8 Total impact
-
Article: A new typing technique for the Rickettsiales Ehrlichia ruminantium: multiple-locus variable number tandem repeat analysis.
[show abstract] [hide abstract]
ABSTRACT: Ehrlichia ruminantium (ER) is a member of the order Rickettsiales transmitted by Amblyomma ticks. This obligatory intracellular bacterium is the causative agent of a fatal disease in ruminants, named heartwater. It represents a constraint on breeding development in sub-Saharan Africa and in the Caribbean. The genetic diversity of the strains of ER, which could be a limiting factor to obtain effective vaccines, needs to be better characterized. For this purpose, we developed a molecular typing technique based on the polymorphism of variable number tandem repeat (VNTR) sequences, MLVA (multiple locus VNTR analysis). Eight (out of 21) VNTR candidates were validated using 17 samples representing a panel of ER strains from different geographical origins from West, South Africa, and Caribbean areas and in ER infected ticks and goat tissues. This result demonstrated the ability of these VNTRs to type a wide range of strains. The stability of the selected VNTR markers was very good, at the time scale needed for epidemiological purposes: in particular, no difference in the VNTR profiles was observed between virulent and attenuated strains (for Gardel and Senegal strains) and between strains (Gardel and Blonde strains) isolated in the same area 19years apart. We validated the strong discriminatory power of MLVA for ER and found a high level of polymorphism between the available strains, with 10 different profiles out of 13 ER strains. The MLVA scheme described in this study is a rapid and efficient molecular typing tool for ER, which allows rapid and direct typing of this intracellular pathogen without preliminary culture and gives reliable results that can be used for further epidemiological studies.Journal of microbiological methods 11/2011; 88(2):205-11. · 2.43 Impact Factor -
Article: Global gene expression profiling of Ehrlichia ruminantium at different stages of development.
[show abstract] [hide abstract]
ABSTRACT: Ehrlichia ruminantium (ER), the causative agent of heartwater on ruminants, is an obligate intracellular bacterium transmitted by ticks of the genus Amblyomma. Previous studies have shown that early stages of development may be critical for Ehrlichia pathogenicity. To gain insights into the biology of intracellular ER, we determined the genome-wide transcriptional profile of ER replicating inside bovine aortic endothelial cells using DNA microarrays. At intermediate and late stages of infection (reticulate and elementary bodies, respectively), a total of 54 genes were differentially expressed. Among them, we measured by q-RTPCR the overexpression of 11 of 14 genes. A number of genes involved in metabolism, nutrient exchange, and defense mechanisms, including those involved in resistance to oxidative stress, were significantly induced in ER reticulate bodies. This is consistent with the oxidative stress condition and nutrient starvation that seem to occur in Ehrlichia-containing vacuoles. During the lysis stage of development, when ER is infectious, we showed the overexpression of a transcription factor, dksA, which is also known to induce virulence in other pathogens such as Salmonella typhimurium. Our results suggest a possible role of these genes in promoting ER development and pathogenicity.FEMS Immunology & Medical Microbiology 11/2011; 64(1):66-73. · 2.44 Impact Factor -
Article: Efficiency of inactivated vaccines against heartwater in Burkina Faso: impact of Ehrlichia ruminantium genetic diversity.
[show abstract] [hide abstract]
ABSTRACT: In order to identify the appropriate strains to use in vaccination trials against heartwater in Burkina Faso, the protective effect of Gardel and Welgevonden strains was assessed against local strains on sheep vaccinated by infection-and-treatment method: Gardel protected significantly against Burkina Faso strains tested (survival rate 59% for immunised sheep vs 13% for control sheep) while Welgevonden did not (survival rate 45% for immunised sheep vs 25% for control sheep). The efficacy of the ISA50 inactivated vaccine, produced under industrial process, was evaluated in sheep during field challenges in two successive years. During year 1, there was a limited protective effect of the Gardel vaccine with 65% of survival rate for the vaccinated group compared to 49% for the control group (N=153, p=0.053). During year 2, the vaccine containing Gardel and a local strain gave an increased protective effect compared to the first trial: 72% of the vaccinated animals survived compared to 47% of the naïve animals (N=173, p<0.001). There was an important genetic diversity of strains in the field with detection of 11 different map1 genotypes in brains from control and vaccinated sheep post mortem. Map1 genotyping of strains detected in brains from control sheep showed that genotype distribution varied according to time and study areas, which could explain the difference in efficacy of the vaccine.Vaccine 06/2010; 28(29):4573-80. · 3.77 Impact Factor -
Article: Mining the genetic diversity of Ehrlichia ruminantium using map genes family.
[show abstract] [hide abstract]
ABSTRACT: Understanding bacterial genetic diversity is crucial to comprehend pathogenesis. Ehrlichia ruminantium (E. ruminantium), a tick-transmitted intracellular bacterial pathogen, causes heartwater disease in ruminants. This model rickettsia, whose genome has been recently sequenced, is restricted to neutrophils and reticulo-endothelial cells of its mammalian host and to the midgut and salivary glands of its vector tick. E. ruminantium harbors a multigene family encoding for 16 outer membrane proteins including MAP1, a major antigenic protein. All the 16 map paralogs are expressed in bovine endothelial cells and some are specifically translated in the tick or in the mammalian host. In this study, we carried out phylogenetic analyses of E. ruminantium using sequences of 6 MAP proteins, MAP1, MAP1-2, MAP1-6, MAP1-5, MAP1+1 and MAP1-14, localized either in the center or at the borders of the map genes cluster. We show that (i) map1 gene is a good tool to characterize the genetic diversity among Africa, Caribbean islands and Madagascar strains including new emerging isolates of E. ruminantium; (ii) the different map paralogs define different genotypes showing divergent evolution; (iii) there is no correlation between all MAP genotypes and the geographic origins of the strains; (iv) The genetic diversity revealed by MAP proteins is conserved whatever is the scale of strains sampling (village, region, continent) and thus was not related to the different timing of strains introduction, i.e. continuous introduction of strains versus punctual introduction (Africa versus Caribbean islands). These results provide therefore a significant advance towards the management of E. ruminantium diversity. The differential evolution of these paralogs suggests specific roles of these proteins in host-vector-pathogen interactions that could be crucial for developing broad-spectrum vaccines.Veterinary Parasitology 09/2009; 167(2-4):187-95. · 2.58 Impact Factor -
Article: Amblyomma variegatum ticks and heartwater on three Caribbean Islands.
[show abstract] [hide abstract]
ABSTRACT: Amblyomma variegatum tick infestation, tick infection by Ehrlichia ruminantium (ER), and ER genetic diversity were studied in the Caribbean Islands of Guadeloupe, Marie-Galante, and Antigua between 2003 and 2005. Nested PCR for pCS20 was used to detect ER, while ER strains were characterized by sequencing or by restriction fragment length polymorphism (RFLP) profiles of map-1 PCR products. In 2003 in Guadeloupe, the prevalence of tick-infested herds was 35.6%. In Marie-Galante 79.1% of herds in 2003 and 73.8% in 2005 were infested, while only an average of 2.2% of the herds were infected in Antigua between this same period. In Marie-Galante, 19.1% of ticks were ER positive, and ER-infected ticks were found in 33.3% of the herds. In Antigua only 5.8% of the ticks were ER positive. High ER tick infection rate combined with a very high level of tick infestation highlight the risk of heartwater in Marie-Galante and Guadeloupe more than in Antigua. The three islands still represent a reservoir for tick and heartwater in the Caribbean. Nine different African and Caribbean map-1 ER genotypes were identified. This diversity was observed even in restricted areas, and identical map-1 genotypes were observed on all three islands. This high genetic diversity of ER strains suggests that there was a simultaneous introduction of several strains from African countries into the Caribbean region.Annals of the New York Academy of Sciences 01/2009; 1149:191-5. · 3.15 Impact Factor -
Article: Innovative approach for transcriptomic analysis of obligate intracellular pathogen: selective capture of transcribed sequences of Ehrlichia ruminantium.
[show abstract] [hide abstract]
ABSTRACT: Whole genome transcriptomic analysis is a powerful approach to elucidate the molecular mechanisms controlling the pathogenesis of obligate intracellular bacteria. However, the major hurdle resides in the low quantity of prokaryotic mRNAs extracted from host cells. Our model Ehrlichia ruminantium (ER), the causative agent of heartwater, is transmitted by tick Amblyomma variegatum. This bacterium affects wild and domestic ruminants and is present in Sub-Saharan Africa and the Caribbean islands. Because of its strictly intracellular location, which constitutes a limitation for its extensive study, the molecular mechanisms involved in its pathogenicity are still poorly understood. We successfully adapted the SCOTS method (Selective Capture of Transcribed Sequences) on the model Rickettsiales ER to capture mRNAs. Southern Blots and RT-PCR revealed an enrichment of ER's cDNAs and a diminution of ribosomal contaminants after three rounds of capture. qRT-PCR and whole-genome ER microarrays hybridizations demonstrated that SCOTS method introduced only a limited bias on gene expression. Indeed, we confirmed the differential gene expression between poorly and highly expressed genes before and after SCOTS captures. The comparative gene expression obtained from ER microarrays data, on samples before and after SCOTS at 96 hpi was significantly correlated (R2 = 0.7). Moreover, SCOTS method is crucial for microarrays analysis of ER, especially for early time points post-infection. There was low detection of transcripts for untreated samples whereas 24% and 70.7% were revealed for SCOTS samples at 24 and 96 hpi respectively. We conclude that this SCOTS method has a key importance for the transcriptomic analysis of ER and can be potentially used for other Rickettsiales. This study constitutes the first step for further gene expression analyses that will lead to a better understanding of both ER pathogenicity and the adaptation of obligate intracellular bacteria to their environment.BMC Molecular Biology 01/2009; 10:111. · 2.86 Impact Factor -
Article: Amblyomma variegatum in cattle in Marie Galante, French Antilles: prevalence, control measures, and infection by Ehrlichia ruminantium.
[show abstract] [hide abstract]
ABSTRACT: We report Marie Galante as one of the Caribbean islands most heavily infested by the tropical bont tick (TBT) Amblyomma variegatum which is associated with two major diseases of ruminants: heartwater and dermatophilosis. In 2005, a survey was undertaken to assess the prevalence of TBT infestation in cattle, the prevalence of Ehrlichia ruminantium infection in TBTs, and the tick control measures implemented by livestock owners. A random sample of 195 cattle herds out of 1885 recorded on the island was investigated by thoroughly counting adult ticks on each animal and filling a questionnaire. A randomly collected sample of 136 TBTs was tested for infection by E. ruminantium by pCS20 nested PCR. Cattle herd prevalence (hp) was 73.8% for infestation by at least one TBT, 17.9% for infestation by at least one engorged female TBT, and 8.2% for clinical dermatophilosis. Cattle individual prevalence was 42.3% for infestation by at least one TBT, 6.6% for infestation by at least one engorged female TBT, and 2.2% for clinical dermatophilosis. The minimum, maximum and average numbers of TBTs per infested animal were, respectively 1, 108 and 11.5. Prevalence of TBT infection by E. ruminantium was 19.1%. No significant difference in herd prevalence was found among parishes or among ecological zones. For cattle owners treating against ticks (97.9% of all owners), all used aspersion of amitraz and herd prevalence was significantly different among those treating every 1-2-week (hp=69.6%, n=148), and less often than every 2-week (hp=88.6%, n=35) (P=0.031). Of the 42 herd subunits treated less than 4 days before the survey, 27 (64%) were infested with at least one TBT, and 6 (14%) with at least one engorged female TBT. These results indicate a high level of TBT infestation in Marie Galante, the inefficacy of tick treatments currently performed, and the need for an improved tick control strategy. Persisting high levels of infestation in Marie Galante threaten the success of on-going TBT eradication programs in the Caribbean because TBT can spread through migrating birds and trade of animals or of animal hides to other islands and potentially the American continent.Veterinary Parasitology 06/2008; 153(3-4):338-46. · 2.58 Impact Factor -
Article: MLST scheme of Ehrlichia ruminantium: Genomic stasis and recombination in strains from Burkina-Faso
[show abstract] [hide abstract]
ABSTRACT: Heartwater, caused by the intracellular bacterium Ehrlichia ruminantium, is a major tick-borne disease of livestock in Africa also introduced in the Caribbean. The main problem encountered with the control of this disease is the lack of efficient vaccine in the field. This is thought to be related to the high genetic diversity of strains circulating in a same area. A set of eight circulating strains was isolated from a herd of cows in a small locality in Burkina-Faso and analyzed along with two reference strains, i.e. ERGA and ERWO, for which full-length genome was available. A MLST analysis was developed based on the genes gltA, groEL, lepA, lipA, lipB, secY, sodB and sucA. Phylogeny analysis was conducted both on concatenated MLST loci and on each individual locus. This showed differing phylogenies for each individual target gene. Most of the recorded polymorphism was borne by three strains: 331, 469 and 623. The neutrality hypothesis could not be rejected. Recombination and linkage disequilibrium were shown to have occurred. A core of seven strains displayed little polymorphism and signs of most likely ancient recombination events. The two reference strains, one from the Caribbean separated from west African strains three centuries ago and another one isolated in South Africa, were very closely related to the core strains whereas the three differing strains displayed recombination and most of the parcimony informative sites. These data suggest that some strains are in genomic stasis, as expected for intracellular parasites, while others emerge in the same area with DNA polymorphism. This work also shows that the MLST scheme developed can discriminate between these two kinds of strains.Infection, Genetics and Evolution.
Top co-authors
Top Journals
Institutions
-
2010
-
Centre international de recherche-développement sur l'elevage en zone subhumide (CIRDES)
Bobo-Dioulasso, Hauts-Bassins, Burkina Faso
-
-
2009
-
Cirad - La recherche agronomique pour le développement
Montpellier, Languedoc-Roussillon, France
-
