Attachment of Coxiella burnetii to the zona pellucida of in vitro produced goat embryos

Autor: Joanna Maria Gonçalves Souza-Fabjan, Laurence Dubreil, E. Rousset, A. Alsaleh, Pascal Mermillod, Annie Rodolakis, C. Roux, J.F. Bruyas, Francis Fieni, J.L. Pellerin
Přispěvatelé: Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), UPSP 5301, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Infectiologie et Santé Publique (UMR ISP), Institut National de la Recherche Agronomique (INRA)-Université de Tours (UT), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Physiopathologie Animale et bioThérapie du muscle et du système nerveux (PAnTher), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Université de Tours
Rok vydání: 2017
Předmět:
0301 basic medicine
Conventional PCR
Bacterial Adhesion
Immunolabeling
Food Animals
Biologie de la reproduction
Small Animals
Zona pellucida
In-vitro-produced embryos
Reproductive Biology
Microscopy
Confocal
biology
production in vitro
Goats
Embryo
Embryo transfer
3. Good health
Real-time polymerase chain reaction
medicine.anatomical_structure
Coxiella burnetii
embryonic structures
Veterinary medicine and animal Health
bactérie pathogène
Autre (Sciences du Vivant)
DNA
Bacterial
[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT]
animal structures
chèvre
microscopie confocale
030106 microbiology
Andrology
03 medical and health sciences
medicine
Animals
caprin
Zona Pellucida
[SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health
Equine
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology
biology.organism_classification
bacterial infections and mycoses
Embryo
Mammalian
Virology
Real-time PCR
Confocal microscopy
In vitro
Médecine vétérinaire et santé animal
zone pellucide
bacteria
Animal Science and Zoology
Bacteria
Zdroj: Theriogenology
Theriogenology, Elsevier, 2018, 106, pp.259-264. ⟨10.1016/j.theriogenology.2017.10.033⟩
Theriogenology (106), 259-264. (2018)
ISSN: 1879-3231
0093-691X
DOI: 10.1016/j.theriogenology.2017.10.033⟩
Popis: International audience; Previous work demonstrated that after infection of in vivo derived caprine embryos, Coxiella burnetti (C. burnetii) showed a strong tendency to adhere to the zona pellicida (ZP). To investigate the risk of C. burnetii transmission via embryo transfer of in vitro-produced goat embryos the aim of this study was, (i) to evaluate the ability of C. burnetii to adhere to the intact zona pellicida of in vitro-produced goat embryos and to determine by confocal microscopy the location of the bacteria, (ii) to test the efficacy of IETS recommended rules for the washing of bovine embryos to eliminate C. burnetii, One hundred ZP-intact caprine embryos, produced in vitro, at the 8 to 16 cell stage, were randomly divided into 11 batches of eight to nine embryos. Nine batches were incubated for 18 h with 109 Coxiella/ml of CbB1 strain (IASP, INRA Tours). The embryos then were recovered and washed in batches in 10 successive baths following the IETS guidelines. In parallel, two batches of embryos were subjected to similar procedures but without exposure to C. burnetii, to serve as the control group. One of the nine batches of infected embryos and one of the two non-infected control batches were separated to perform immunolabeling to locate the bacteria. C. burnetii DNA was detected by C-PCR in all eight batches of infected embryos after 10 successive washings. However, bacterial DNA was not detected in the embryo control batch. The first five washing media of the infected group were consistently found to be positive and Coxiella DNA was detected in the wash bath up to the 10th wash for two batches. After immunolabeling, the observation of embryos under confocal microscopy allowed C. burnetti to be found on the external part of the zona pellucida without deep penetration. This study clearly demonstrates that C. burnetii, after in vitro infection at 109 Coxiella/ml, stick strongly to the external part of the zona pellucida of in vitro produced caprine embryos without deap penetration and that the 10 washings protocol recommended by IETS to eliminate the pathogenic agents of bovine embryos is unable to eliminate these bacteria from in vitro-produced goat embryo.
Databáze: OpenAIRE
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