title: Anaplasma platys-like strains in ruminants from Tunisia.
abstract: Molecular diagnosis of Anaplasma platys and related strains (A. platys-like) in carnivores and ruminants is challenging due to co-infections with cross-reacting strains, and require post-amplification sequencing of the hemi-nested PCR products traditionally generated by targeting the groEL gene. In this study, a Restriction Enzyme Fragment Length Polymorphism (RFLP) assay coupled to hemi-nested groEL PCR was developed to discriminate among A. platys and genetically related strains. This novel approach was used for investigating A. platys-like infection in 963 domesticated ruminants (241 goats, 355 sheep, and 367 cattle) from 22 delegations located in North Tunisia. Overall prevalence rates of A. platys-like were 22.8, 11, and 3.5% in goats, sheep, and cattle, respectively. Alignment, identity comparison, and phylogenetic analysis of the groEL sequence variants obtained in this study confirmed RFLP data suggesting that Tunisian ruminants are infected by novel unclassified Anaplasma strains genetically related to A. platys. Compared to sequencing, RFLP assay allows fast detection of A. platys and A. platys-like pathogens in the same sample and has a potential value especially when screening ticks, cats and ruminants, which can be a common host for these two bacteria. This newly developed molecular technique would provide valuable molecular tool for epidemiological studies related to A. platys as well as remove concern over specificity of serological and molecular methods routinely used to identify diverse Anaplasma strains and species in wild and domestic ruminants.
authors.affiliation: Laboratoire de Microbiologie, Ecole Nationale de Médecine Vétérinaire, Institution de la Recherche et de l'Enseignement Supérieur Agricoles, Université de La Manouba, Sidi Thabet, Tunisia.
authors.forename: Mourad
authors.initials: M
authors.lastname: Ben Said
authors.affiliation: Laboratoire de Microbiologie, Ecole Nationale de Médecine Vétérinaire, Institution de la Recherche et de l'Enseignement Supérieur Agricoles, Université de La Manouba, Sidi Thabet, Tunisia.
authors.forename: Hanène
authors.initials: H
authors.lastname: Belkahia
authors.affiliation: Laboratoire de Microbiologie, Ecole Nationale de Médecine Vétérinaire, Institution de la Recherche et de l'Enseignement Supérieur Agricoles, Université de La Manouba, Sidi Thabet, Tunisia.
authors.forename: Narjesse
authors.initials: N
authors.lastname: El Mabrouk
authors.affiliation: Laboratoire de Microbiologie, Ecole Nationale de Médecine Vétérinaire, Institution de la Recherche et de l'Enseignement Supérieur Agricoles, Université de La Manouba, Sidi Thabet, Tunisia.
authors.forename: Mariem
authors.initials: M
authors.lastname: Saidani
authors.affiliation: Dipartimento di Medicina Veterinaria, Università degli Studi di Sassari, Sassari, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Alberti
authors.affiliation: Dipartimento di Medicina Veterinaria, Università degli Studi di Sassari, Sassari, Italy.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Zobba
authors.affiliation: Laboratoire de Microbiologie, Ecole Nationale de Médecine Vétérinaire, Institution de la Recherche et de l'Enseignement Supérieur Agricoles, Université de La Manouba, Sidi Thabet, Tunisia.
authors.forename: Amal
authors.initials: A
authors.lastname: Cherif
authors.affiliation: Laboratoire de Microbiologie, Ecole Nationale de Médecine Vétérinaire, Institution de la Recherche et de l'Enseignement Supérieur Agricoles, Université de La Manouba, Sidi Thabet, Tunisia.
authors.forename: Tarek
authors.initials: T
authors.lastname: Mahjoub
authors.affiliation: Laboratoire d'Entomologie, Institut Pasteur de Tunis, Tunisia.
authors.forename: Ali
authors.initials: A
authors.lastname: Bouattour
authors.affiliation: Laboratoire de Microbiologie, Ecole Nationale de Médecine Vétérinaire, Institution de la Recherche et de l'Enseignement Supérieur Agricoles, Université de La Manouba, Sidi Thabet, Tunisia. Electronic address: lilia_messadi@yahoo.fr.
authors.forename: Lilia
authors.initials: L
authors.lastname: Messadi
mesh_terms: D000709:Anaplasma / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D000712:Anaplasmosis / Q000453:epidemiology* / Q000382:microbiology; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000453:epidemiology* / Q000382:microbiology; D018834:Chaperonin 60 / Q000235:genetics*; D015870:Gene Expression; D015511:Goat Diseases / Q000453:epidemiology* / Q000382:microbiology; D006041:Goats / Q000382:microbiology; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D016415:Sequence Alignment; D012756:Sheep / Q000382:microbiology; D012757:Sheep Diseases / Q000453:epidemiology* / Q000382:microbiology; D014416:Tunisia / Q000453:epidemiology
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60
keywords: Anaplasma platys-like strains; Novel genetic variants; Ruminants; Specific detection; Tunisia; groEL gene