Abstract

Introduction: Brucellosis is a zoonotic disease that has a significant economic, social and public health impact in many parts of the world. The causative agents are members of the genus Brucella currently comprising 11 species and with an expanding known host range in recent years. Case presentation: One of a pair of White’s tree frogs (Litoria caerulea) developed skin lesions from which a pure growth of a haemolytic organism was obtained. The isolate was identified as Brucella melitensis by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry, although the colony morphology was inconsistent with this identification. Applying the classical biotyping approach used to subdivide members of the genus Brucella, the isolate did not correspond to any known Brucella sp. However, PCR targeting of genes specific for members of the genus Brucella was strongly positive and 16S rRNA gene sequencing revealed a close relationship with extant Brucella spp. In order to place the isolate more accurately, a multilocus sequencing approach was applied, which confirmed that the isolate represented a novel member of the emerging ‘atypical’ Brucella group, which includes isolates from human disease, from rodents and, more recently, reported isolations from frogs in Germany. Conclusion: This case represents the first report of isolation of a Brucella sp. from frogs outside Germany and suggests that these isolates may be widespread. Whilst there is no evidence to date that these isolates represent a zoonotic threat, the association of other ‘atypical’ Brucella sp. with human disease suggests that appropriate measures should be taken to avoid unnecessary contact with potentially infected amphibians until the zoonotic potential of this group is better understood.

Highlights

  • Brucellosis is a zoonotic disease that has a significant economic, social and public health impact in many parts of the world

  • We add to this picture describing a case of isolation of Abbreviations: APHA, Animal and Plant Health Agency; MALDI-TOF MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

  • Whilst this isolate would be excluded as Brucella by conventional phenotyping approaches, molecular analysis confirmed that the isolate belongs to the rapidly expanding group of ‘atypical’ Brucella spp

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Summary

Introduction

Brucellosis is a zoonotic disease with a significant global impact. Of the 11 recognized species, Brucella melitensis, Brucella abortus and Brucella suis, causative agents of small ruminant, bovine and swine brucellosis, respectively, are considered the most significant animal and human pathogens. The isolate was examined by a slide agglutination test using unabsorbed Brucella antiserum and showed strong agglutination This test is considered to lack specificity, and the gold standard for identification is a biotyping approach conventionally used to identify Brucella isolates to the species and biovar levels (Whatmore, 2009). Phylogenetic placement based on concatenated sequence data, excluding a single gene that does not amplify from B. inopinata, was performed, comparing the isolate with isolates representing the known genetic diversity of members of the genus Brucella (Fig. 2b). These included both ‘core’ Brucella spp., comprising all the classically described major pathogenic species, as well as. Whole-genome sequencing currently in progress should provide a route to understanding the structure of UK8/14 LPS

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