Silvio Spicic1, Maja Zdelar-Tuk2, Claire Ponsart3, Rene S Hendriksen4, Irena Reil2, Guillaume Girault3, Pimlapas Leekitcharoenphon4, Vesna Rukavina5, Martina Rubin6, Luca Freddi3, Sanja Duvnjak2. 1. Department of Bacteriology and Parasitology, Laboratory for Bacterial Zoonosis and Molecular Diagnostics of Bacterial Diseases, Croatian Veterinary Institute, Savska street 143, Zagreb, Croatia. spicic@veinst.hr. 2. Department of Bacteriology and Parasitology, Laboratory for Bacterial Zoonosis and Molecular Diagnostics of Bacterial Diseases, Croatian Veterinary Institute, Savska street 143, Zagreb, Croatia. 3. French Agency for Food, Environmental & Occupational Health Safety (ANSES), Bacterial Zoonoses Unit - Animal Health Laboratory, National & OIE/FAO Animal Brucellosis Reference Laboratory, EU Reference Laboratory for Brucellosis, 14 rue Pierre et Marie Curie, Maisons-Alfort, Cedex, France. 4. Technical University of Denmark, National Food Institute, Research Group for Genomic Epidemiology, Kemitorvet, Building 204, 2800, Kgs. Lyngby, Denmark. 5. Ministry of Agriculture, Veterinary and Food Safety Directorate, Veterinary Office Sisak, Branch Office Glina, Trg bana Josipa Jelačića 2, 44 400, Glina, Croatia. 6. Ministry of Agriculture, Veterinary and Food Safety Directorate, Department for Veterinary Epidemiology, Planinska 2a, 10000, Zagreb, Croatia.
Abstract
BACKGROUND: A novel Brucella strain closely related to Brucella (B.) melitensis biovar (bv) 3 was found in Croatian cattle during testing within a brucellosis eradication programme. CASE PRESENTATION: Standardised serological, brucellin skin test, bacteriological and molecular diagnostic screening for Brucella infection led to positive detection in one dairy cattle herd. Three isolates from that herd were identified to species level using the Bruce ladder method. Initially, two strains were typed as B. melitensis and one as B. abortus, but multiplex PCR based on IS711 and the Suis ladder showed that all of them to belong to B. melitensis, and the combination of whole-genome and multi-locus sequencing as well as Multi-Locus Variable numbers of tandem repeats Analysis (MLVA) highlighted a strong proximity within the phylogenetic branch of B. melitensis strains previously isolated from Croatia, Albania, Kosovo and Bosnia and Herzegovina. Two isolates were determined to be B. melitensis bv. 3, while the third showed a unique phylogenetic profile, growth profile on dyes and bacteriophage typing results. This isolate contained the 609-bp omp31 sequence, but not the 723-bp omp31 sequence present in the two isolates of B. melitensis bv. 3. CONCLUSIONS: Identification of a novel Brucella variant in this geographic region is predictable given the historic endemicity of brucellosis. The emergence of a new variant may reflect a combination of high prevalence among domestic ruminants and humans as well as weak eradication strategies. The zoonotic potential, reservoirs and transmission pathways of this and other Brucella variants should be explored.
BACKGROUND: A novel Brucella strain closely related to Brucella (B.) melitensis biovar (bv) 3 was found in Croatian cattle during testing within a brucellosis eradication programme. CASE PRESENTATION: Standardised serological, brucellin skin test, bacteriological and molecular diagnostic screening for Brucella infection led to positive detection in one dairy cattle herd. Three isolates from that herd were identified to species level using the Bruce ladder method. Initially, two strains were typed as B. melitensis and one as B. abortus, but multiplex PCR based on IS711 and the Suis ladder showed that all of them to belong to B. melitensis, and the combination of whole-genome and multi-locus sequencing as well as Multi-Locus Variable numbers of tandem repeats Analysis (MLVA) highlighted a strong proximity within the phylogenetic branch of B. melitensis strains previously isolated from Croatia, Albania, Kosovo and Bosnia and Herzegovina. Two isolates were determined to be B. melitensis bv. 3, while the third showed a unique phylogenetic profile, growth profile on dyes and bacteriophage typing results. This isolate contained the 609-bp omp31 sequence, but not the 723-bp omp31 sequence present in the two isolates of B. melitensis bv. 3. CONCLUSIONS: Identification of a novel Brucella variant in this geographic region is predictable given the historic endemicity of brucellosis. The emergence of a new variant may reflect a combination of high prevalence among domestic ruminants and humans as well as weak eradication strategies. The zoonotic potential, reservoirs and transmission pathways of this and other Brucella variants should be explored.
Authors: I López-Goñi; D García-Yoldi; C M Marín; M J de Miguel; P M Muñoz; J M Blasco; I Jacques; M Grayon; A Cloeckaert; A C Ferreira; R Cardoso; M I Corrêa de Sá; K Walravens; D Albert; B Garin-Bastuji Journal: J Clin Microbiol Date: 2008-08-20 Impact factor: 5.948
Authors: Ignacio López-Goñi; David García-Yoldi; Clara M Marín; María J de Miguel; Elías Barquero-Calvo; Caterina Guzmán-Verri; David Albert; Bruno Garin-Bastuji Journal: Vet Microbiol Date: 2011-07-02 Impact factor: 3.293
Authors: Philippe Le Flèche; Isabelle Jacques; Maggy Grayon; Sascha Al Dahouk; Patrick Bouchon; France Denoeud; Karsten Nöckler; Heinrich Neubauer; Laurence A Guilloteau; Gilles Vergnaud Journal: BMC Microbiol Date: 2006-02-09 Impact factor: 3.605