Abstract

BackgroundChlamydia abortus and Chlamydia psittaci are important pathogens of livestock and avian species, respectively. While C. abortus is recognized as descended from C. psittaci species, there is emerging evidence of strains that are intermediary between the two species, suggesting they are recent evolutionary ancestors of C. abortus. Such strains include C. psittaci strain 84/2334 that was isolated from a parrot. Our aim was to classify this strain by sequencing its genome and explore its evolutionary relationship to both C. abortus and C. psittaci.ResultsIn this study, methods based on multi-locus sequence typing (MLST) of seven housekeeping genes and on typing of five species discriminant proteins showed that strain 84/2334 clustered with C. abortus species. Furthermore, whole genome de novo sequencing of the strain revealed greater similarity to C. abortus in terms of GC content, while 16S rRNA and whole genome phylogenetic analysis, as well as network and recombination analysis showed that the strain clusters more closely with C. abortus strains. The analysis also suggested a closer evolutionary relationship between this strain and the major C. abortus clade, than to two other intermediary avian C. abortus strains or C. psittaci strains. Molecular analyses of genes (polymorphic membrane protein and transmembrane head protein genes) and loci (plasticity zone), found in key virulence-associated regions that exhibit greatest diversity within and between chlamydial species, reveal greater diversity than present in sequenced C. abortus genomes as well as similar features to both C. abortus and C. psittaci species. The strain also possesses an extrachromosomal plasmid, as found in most C. psittaci species but absent from all sequenced classical C. abortus strains.ConclusionOverall, the results show that C. psittaci strain 84/2334 clusters very closely with C. abortus strains, and are consistent with the strain being a recent C. abortus ancestral species. This suggests that the strain should be reclassified as C. abortus. Furthermore, the identification of a C. abortus strain bearing an extra-chromosomal plasmid has implications for plasmid-based transformation studies to investigate gene function as well as providing a potential route for the development of a next generation vaccine to protect livestock from C. abortus infection.

Highlights

  • The family Chlamydiaceae comprises a group of obligate intracellular Gram-negative bacteria that are responsible for a broad range of infections in non-human mammals, birds and humans [1]

  • Overall, the results show that C. psittaci strain 84/2334 clusters very closely with C. abortus strains, and are consistent with the strain being a recent C. abortus ancestral species

  • Phylogenetic analysis of the 16S rRNA sequences produced a similar tree structure to that obtained by multi-locus sequence typing (MLST) analysis (Additional file 3), the only exception being C. buteonis strain RSHA which clusters with C. abortus species, while ribosomal MLST based on 53 genes encoding the bacterial ribosome protein subunits [34] predicted the species taxon of strain 84/2334 as C. abortus with 100% support (Additional file 4)

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Summary

Introduction

The family Chlamydiaceae comprises a group of obligate intracellular Gram-negative bacteria that are responsible for a broad range of infections in non-human mammals, birds and humans [1]. Since 1999 the family has undergone a number of taxonomic reclassifications at the genus and species levels based on sequence analysis of the 16S and 23S rRNA genes, as well as taking into account biological differences [2,3,4,5]. Current classification within this family recognises a single genus, Chlamydia, and 14 species (Chlamydia abortus, Chlamydia avium, Chlamydia buteonis, Chlamydia caviae, Chlamydia felis, Chlamydia gallinacea, Chlamydia muridarum, Chlamydia pecorum, Chlamydia pneumoniae, Chlamydia poikilothermis, Chlamydia psittaci, Chlamydia serpentis, Chlamydia suis and Chlamydia trachomatis) [6,7,8], plus a further four Candidatus species (Ca. Chlamydia ibidis, Ca. Chlamydia corallus, Ca. Chlamydia sanzinia and Ca. Chlamydia testudinis) [9,10,11,12]. Our aim was to classify this strain by sequencing its genome and explore its evolutionary relationship to both C. abortus and C. psittaci

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