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1. Swab cloths as a tool for revealing environmental contamination by Q fever in ruminant farms

Author

Pauline Carrie, Séverine Barry, Elodie Rousset, Patrick Gasqui, Renée de Crémoux, Claire A.M. Becker, Didier Calavas, Emmanuelle Gilot-Fromont, Carole Sala, Elsa Jourdain, Anne Bronner, Jean-Baptiste Perrin, Kristel Gache, Unité Mixte de Recherche d'Épidémiologie des maladies Animales et zoonotiques (UMR EPIA), Institut National de la Recherche Agronomique (INRA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Université de Lyon, VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Unité Fièvre Animale Q (NRL Fièvre Q), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de Lyon [ANSES], Bureau de Santé Animale, DGAL, Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Mycoplasmoses des Ruminants - UMR (MYCO), Université de Lyon-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Fédération nationale des Groupements de Défense Sanitaire (GDS France), Institut de l'élevage, Inst Elevage, Unité Fièvre Animale Q (Fièvre Q), Laboratoire de Sophia Antipolis, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Université de Lyon-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut de l'élevage (IDELE), Épidémiologie des Maladies Animales et Zoonotiques - UMR 346 (EPIA), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Unité de recherche d'Épidémiologie Animale (UEA), Institut National de la Recherche Agronomique (INRA), Université de Montréal [Montréal], AFSSA Lyon, Unité Epidémiologie, Laboratoire de Lyon, Bureau de la Santé Animale, Direction Générale de l'Alimentation (DGAL), National animal health farmers’organisation, and GDS

Subjects
Veterinary medicine, Farms, 040301 veterinary sciences, animal diseases, Cattle Diseases, Sheep Diseases, Q fever, Real-Time Polymerase Chain Reaction, 0403 veterinary science, 03 medical and health sciences, Ruminant, Pregnancy, Zoonoses, medicine, Environmental Microbiology, Small ruminant, Animals, Humans, Epidemics, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 2. Zero hunger, 0303 health sciences, Coxiella burnetii DNA, Goat Diseases, Sheep, General Veterinary, General Immunology and Microbiology, biology, Goats, Outbreak, Dust, 04 agricultural and veterinary sciences, General Medicine, Contamination, bacterial infections and mycoses, medicine.disease, Coxiella burnetii, biology.organism_classification, Housing, Animal, 3. Good health, Herd, bacteria, Cattle, Female, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, Q Fever
Abstract

Q fever is a zoonotic abortive disease of ruminants mostly transmitted by inhalation of aerosols contaminated by Coxiella burnetii. Clusters of cases or even epidemics regularly occur in humans but, to date, there is no consensus about the best way to carry out outbreak investigations in order to identify potential farms at risk. Although environmental samples might be useful during such investigations, there are few baseline data on the presence of C. burnetii in the environment of ruminant farms. We thus investigated dust samples from cattle, sheep and goat farm buildings in order to (a) estimate C. burnetii detection frequency and bacterial loads in the environment, and (b) determine whether this environmental contamination is associated with series of abortions attributed to Q fever. We considered 113 herds with a recent abortive episode potentially related (n = 60) or not (n = 53) to C. burnetii. Dust was sampled using a swab cloth and tested by a quantitative PCR method targeting the IS1111 gene. Coxiella burnetii DNA was detected on 9 of 50 cattle farms, 13 of 19 goat farms and 30 of 40 sheep farms. On 16 cloths, bacterial loads were higher than 108 genome equivalents, levels as high as in infectious materials such as placentas and aborted foetuses. Overall, the probability of detecting C. burnetii DNA was higher on small ruminant farms than cattle farms, in herds suspected of Q fever and in large herds. We conclude that swab cloths are a putative indicator of contamination of ruminant farms by C. burnetii.

Published
2019

2. Monitoring the Decrease in Susceptibility to Ribosomal RNAs Targeting Antimicrobials and Its Molecular Basis in Clinical Mycoplasma bovis Isolates over Time

Author

Claire A.M. Becker, Florence Tardy, Dima Khalil, Mycoplasmoses des Ruminants - UMR (MYCO), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Université de Lyon, and Université de Lyon-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)

Subjects
0301 basic medicine, Microbiology (medical), Mycoplasma bovis, [SDV]Life Sciences [q-bio], 030106 microbiology, Immunology, Cattle Diseases, Oxytetracycline, Drug resistance, Microbial Sensitivity Tests, Biology, Tylosin, medicine.disease_cause, Microbiology, 03 medical and health sciences, chemistry.chemical_compound, Anti-Infective Agents, 23S ribosomal RNA, RNA, Ribosomal, 16S, Genotype, Drug Resistance, Bacterial, medicine, Animals, Mycoplasma Infections, Tilmicosin, mycoplasma, Pharmacology, drug resistance, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, macrolides, Mycoplasma, Ribosomal RNA, tetracyclines, Virology, RNA, Ribosomal, 23S, chemistry, Cattle, France, medicine.drug
Abstract

International audience; Mycoplasma bovis is considered an emerging threat to bovine production in industrialized countries. Its control depends on good husbandry and efficient chemotherapy practices. In France, clinical isolates collected after 2009 showed a drastic loss of susceptibility to most antimicrobials when compared with isolates collected in 1978-1979. The aim of the present study was to analyze the molecular mechanisms underlying the shift toward resistance to macrolides and tetracyclines and to assess whether the clinical origin of the isolates or their molecular subtypes could have influenced their pattern of evolution. We demonstrated that all M. bovis isolates collected as early as 2000 should already be considered resistant to tylosin, tilmicosin, and oxytetracycline, whatever the associated clinical signs. The shift toward resistance happened earlier for oxytetracycline and more progressively for tylosin/tilmicosin. Isolates belonging to the major subtype ST2 (n = 40) showed a homogeneous genotype for resistance, with combined alterations of G 748 A and A 2058 G in the 23S rRNA alleles for tylosin/tilmicosin and of A 965 T and A 967 T in the 16S rRNA alleles for oxytetracycline. The genotypes of ST3 or ST1 isolates (n = 9 and 25, respectively) in the process of becoming resistant were more varied. In recent years, the convergence of both ST2 and ST3 isolates toward the same resistance genotype suggests that the corresponding mutations have been selected for providing an appropriate balance between fitness cost and resistance.

Published
2017

3. Identification of three CCp genes in Babesia divergens: Novel markers for sexual stages parasites

Author

Thibaut Larcher, Claire A.M. Becker, Laurence Malandrin, Peter H. David, Emmanuel Bischoff, D. Depoix, Sarah Bonnet, Alain Chauvin, Molécules de Communication et Adaptation des Micro-organismes (MCAM), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), INRA, Region Pays de Loire, Delegation Generale pour l'Armement (DGA) [22120/DSP/SREAF, 04 34 025], 'Fonds dedie Combattre les Maladies Parasitaires' (Sanofi-Aventis/Ministere de l'Enseignement Superieur et de la Recherche), AIP Bioressources, Biologie, Epidémiologie et analyse de risque en Santé Animale (BIOEPAR), Institut National de la Recherche Agronomique (INRA)-École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), Expertise en Anatomie Pathologique (APEX), Physiopathologie Animale et bioThérapie du muscle et du système nerveux (PAnTher), École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Immunologie moléculaire des parasites, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Puces à ADN (Plate-Forme 2) (PF2), Institut Pasteur [Paris] (IP), and This work is part of CB's PhD thesis, which was funded by INRA and Region Pays de Loire. EB was funded by the Délégation Générale pour l’Armement (DGA nos 22120/DSP/SREAF and 04 34 025). The work also received financial support from the 'Fonds dédié Combattre les Maladies Parasitaires' (Sanofi-Aventis/Ministère de l’Enseignement Supérieur et de la Recherche). Sequencing was partially funded by AIP Bioressources.

Subjects
Genetic Markers, EXPRESSION, Plasmodium, Virulence Factors, Sexual stages, TRANSMISSION, In silico, [SDV]Life Sciences [q-bio], Molecular Sequence Data, gametocytes, Protozoan Proteins, Babesia, Genome, 030308 mycology & parasitology, CLONING, 03 medical and health sciences, Babesia divergens, parasitic diseases, Animals, Parasite hosting, Molecular Biology, Gene, 030304 developmental biology, Genetics, 0303 health sciences, Ixodes, biology, PLASMODIUM-FALCIPARUM, Sequence Analysis, DNA, DNA, Protozoan, biology.organism_classification, Virology, GENOME-SEQUENCE, LIFE-CYCLE, Sexual reproduction, PROTEIN DISULFIDE-ISOMERASE, MOSQUITO, HOMOLOG, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, CRYPTOSPORIDIUM-PARVUM, Parasitology, CCp family
Abstract

International audience; Babesia divergens, a tick-borne protozoan parasite of red blood cells, is the main agent of bovine and human babesiosis in Europe. Very few data are available concerning its life cycle and sexual reproduction inside the tick vector, Ixodes ricinus. The aim of this study was to define some markers of the B. divergens sexual stage. An in silico post-genomic approach was used to analyze genomic, transcriptomic and proteomic data and to select specific sexual stage proteins of the related apicomplexan genus Plasmodium. Three proteins, based on sequence identity between the available genomes of Plasmodium and Babesia spp., were chosen, as members of a highly conserved and apicomplexan sexual stages specific protein family (CCp) potentially involved in adhesive functions. Degenerate primers were used to amplify and clone three B. divergens orthologs (bdccp1, bdccp2, and bdccp3) corresponding to newly identified genes in this parasite. The opportunities offered by such markers to study parasite development in its vector are discussed. (C) 2010 Elsevier B.V. All rights reserved.

Published
2010

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