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Rahelinirina S.,Pasteur Institute | Leon A.,Frank Duncombe Laboratory | Harstskeerl R.A.,OIE and National Leptospirosis Reference Center | Sertour N.,Institute Pasteur Paris | And 9 more authors.
PLoS ONE | Year: 2010

Background: Leptospirosis has long been a major public health concern in the southwestern Indian Ocean. However, in Madagascar, only a few, old studies have provided indirect serological evidence of the disease in humans or animals. Methodology/Principal Findings: We conducted a large animal study focusing on small-mammal populations. Five field trapping surveys were carried out at five sites, from April 2008 to August 2009. Captures consisted of Rattus norvegicus (35.8%), R. rattus (35.1%), Mus musculus (20.5%) and Suncus murinus (8.6%). We used microbiological culture, serodiagnosis tests (MAT) and real-time PCR to assess Leptospira infection. Leptospira carriage was detected by PCR in 91 (33.9%) of the 268 small mammals, by MAT in 17 of the 151 (11.3%) animals for which serum samples were available and by culture in 9 of the 268 animals (3.3%). Rates of infection based on positive PCR results were significantly higher in Moramanga (54%), Toliara (48%) and Mahajanga (47.4%) than in Antsiranana (8.5%) and Toamasina (14%) (p = 0.001). The prevalence of Leptospira carriage was significantly higher in R. norvegicus (48.9%), S. murinus (43.5%) and R. rattus (30.8%) than in M. musculus (9.1%) (p<0.001). The MAT detected antibodies against the serogroups Canicola and Icterohaemorrhagiae. Isolates were characterized by serology, secY sequence-based phylogeny, partial sequencing of rrs, multi-locus VNTR analysis and pulsed field gel electrophoresis. The 10 isolates obtained from nine rats were all identified as species L. interrogans serogroup Canicola serovar Kuwait and all had identical partial rrs and secY sequences. Conclusions/Significance: We present here the first direct evidence of widespread leptospiral carriage in small mammals in Madagascar. Our results strongly suggest a high level of environmental contamination, consistent with probable transmission of the infection to humans. This first isolation of pathogenic Leptospira strains in this country may significantly improve the detection of specific antibodies in human cases. © 2010 Rahelinirina et al. Source

Hans A.,Virology Unit | Gaudaire D.,Virology Unit | Manuguerra J.-C.,Institute Pasteur Paris | Leon A.,Frank Duncombe Laboratory | And 6 more authors.
Journal of Clinical Microbiology | Year: 2015

This study shows that an unbiased amplification method applied to equine arteritis virus RNA significantly improves the sensitivity of the real-time reverse transcription-quantitative PCR (RT-qPCR) recommended by the World Organization for Animal Health. Twelve viral RNAs amplified using this method were hybridized on a high-density resequencing microarray for effective viral characterization. Copyright © 2015, American Society for Microbiology. All Rights Reserved. Source

Ramery E.,University of Liege | Fraipont A.,University of Liege | Richard E.A.,University of Liege | Richard E.A.,Frank Duncombe Laboratory | And 6 more authors.
Veterinary Clinical Pathology | Year: 2015

Background: Inflammatory airway disease (IAD) affects performance and well-being of horses. Diagnosis is primarily reached by bronchoalveolar lavage (BAL) cytology which is invasive and requires sedation. Objectives: The purpose of this study was to identify differential gene expression in peripheral blood of horses with IAD using species-specific expression microarrays. Methods: Equine gene expression microarrays were used to investigate global mRNA expression in circulating leukocytes from healthy, IAD-affected, and low-performing Standardbred and endurance horses. Results: Nine genes in Standardbred and 61 genes in endurance horses were significantly differentially regulated (P < .001). These genes were related to inflammation (eg, ALOX15B, PLA2G12B, and PENK), oxidant/antioxidant balance (eg, DUOXA2 and GSTO1-1), and stress (eg, V1aR, GRLF1, Homer-2, and MAOB). All these genes were up-regulated, except down-regulated Homer-2 and MAOB. DUOXA2, ALOX15B, PLA2G12B, MAOB, and GRLF1 expression was further validated by RT-qPCR. An increase in glutathione peroxidase (GPx) activity in heparinized whole blood of IAD-affected Standardbred (P = .0025) and endurance horses (P = .0028) also suggests a deregulation of the oxidant/antioxidant balance. There was good correlation (r = .7354) between BAL neutrophil percentage and whole blood GPx activity in all horses. Conclusions: This study showed that circulating blood cell gene expression reflects inflammatory responses in tissues. Whether any of the genes have potential for diagnostic applications in the future remains to be investigated. Although not specific for IAD, whole blood GPx activity appears to be correlated with BAL neutrophil percentage. This finding should be further assessed by testing a larger number of horses. © 2014 American Society for Veterinary Clinical Pathology. Source

Leon A.,Frank Duncombe Laboratory | Pronost S.,Frank Duncombe Laboratory | Fortier G.,Frank Duncombe Laboratory | Andre-Fontaine G.,Leptospira Medical and Molecular Bacteriology Unit | Leclercq R.,University of Caen Lower Normandy
Journal of Clinical Microbiology | Year: 2010

Fifty-three strains belonging to the pathogenic species Leptospira interrogans and Leptospira kirschneri were analyzed by multilocus sequence analysis. The species formed two distinct branches. In the L. interrogans branch, the phylogenetic tree clustered the strains into three subgroups. Genogroups and serogroups were superimposed but not strictly. Copyright © 2010, American Society for Microbiology. All Rights Reserved. Source

Depecker M.,National School of Engineering in Agricultural and Food Industries | Richard E.A.,Frank Duncombe Laboratory | Pitel P.-H.,Frank Duncombe Laboratory | Fortier G.,Frank Duncombe Laboratory | And 3 more authors.
Veterinary Journal | Year: 2014

The aim of this study was to determine whether the lung side being sampled would significantly influence bronchoalveolar lavage (BAL) cytological profiles and subsequent diagnosis in Standardbred racehorses. One hundred and thirty-eight French Trotters in active training and racing were included in a prospective observational study. BAL was performed using videoendoscopy in both right and left lungs during summer meetings in 2011 (64 horses) and 2012 (74 horses). Cytological data performed 24. h later from right and left lungs were compared and specifically used to classify horses as affected with exercise-induced pulmonary haemorrhage (EIPH), inflammatory airway disease (IAD), or were 'controls'. For IAD, cytological definition was based on two different cut off values.Neutrophil percentages, haemosiderophage percentages and the haemosiderophage/macrophage (H/M) ratios were significantly higher in the right compared to the left lung. Measures of intra-class correlation coefficients revealed a fair agreement between left and right lungs for percentages of mast cells, eosinophils, and for the H/M ratio, and a moderate agreement for neutrophil percentages. Fair to moderate agreements were observed between left and right lungs for the diagnosis of IAD and/or EIPH based on kappa coefficients. When sampling one lung only, the risk of incorrectly classifying a horse as a 'control' increased with the use of the restraint cut-off values for IAD. As BAL from one lung is not representative of the other lung in the same horse, both lungs should be sampled for a better assessment of lung cellularity and for a precise diagnosis of lower airway diseases. © 2013 Elsevier Ltd. Source

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