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To investigate the epidemiological relationship of isolates from different Portuguese geographical regions and to assess the diversity among isolates, the MLVA16 Orsay  assay (panels 1, 2A and 2B) was performed with a collection of 126  Brucella melitensis  (46 human and 80 animal isolates) and 157  B. abortus  field isolates, seven vaccine strains and the representative reference strains of each species. The MLVA16 Orsay  showed a similar high discriminatory power (HGDI 0.972 and 0.902) for both species but panel 1 and 2A markers displayed higher diversity (HGDI 0.693) in  B. abortus  compared to  B. melitensis  isolates (HGDI 0.342). The  B. melitensis  population belong to the “Americas” (17%) and “East Mediterranean” (83%) groups. No isolate belonged to the “West Mediterranean” group. Eighty-five percent of the human isolates (39 in 46) fit in the “East-Mediterranean” group where a single lineage known as MLVA11 genotype 116 is responsible for the vast majority of  Brucella  infections in humans.  B. abortus  isolates formed a consistent group with bv1 and bv3 isolates in different clusters. Four MLVA11 genotypes were observed for the first time in isolates from S. Jorge and Terceira islands from Azores. From the collection of isolates analysed in this study we conclude that MLVA16 Orsay  provided a clear view of  Brucella  spp. population, confirming epidemiological linkage in outbreak investigations. In particular, it suggests recent and ongoing colonisation of Portugal with one  B. melitensis  lineage usually associated with East Mediterranean countries.

MLVA16 Typing of Portuguese Human and Animal Brucella melitensis and Brucella abortus Isolates