Comparative genomics of the dairy isolate Streptococcus macedonicus ACA-DC 198 against related members of the Streptococcus bovis/Streptococcus equinus complex

Abstract

Background: Within the genus Streptococcus, only Streptococcus thermophilus is used as a starter culture in food fermentations. Streptococcus macedonicus though, which belongs to the Streptococcus bovis/Streptococcus equinus complex (SBSEC), is also frequently isolated from fermented foods mainly of dairy origin. Members of the SBSEC have been implicated in human endocarditis and colon cancer. Here we compare the genome sequence of the dairy isolate S. macedonicus ACA-DC 198 to the other SBSEC genomes in order to assess in silico its potential adaptation to milk and its pathogenicity status.

Results: Despite the fact that the SBSEC species were found tightly related based on whole genome phylogeny of streptococci, two distinct patterns of evolution were identified among them. Streptococcus macedonicus, Streptococcus infantarius CJ18 and Streptococcus pasteurianus ATCC 43144 seem to have undergone reductive evolution resulting in significantly diminished genome sizes and increased percentages of potential pseudogenes when compared to Streptococcus gallolyticus subsp. gallolyticus. In addition, the three species seem to have lost genes for catabolizing complex plant carbohydrates and for detoxifying toxic substances previously linked to the ability of S. gallolyticus to survive in the rumen. Analysis of the S. macedonicus genome revealed features that could support adaptation to milk, including an extra gene cluster for lactose and galactose metabolism, a proteolytic system for casein hydrolysis, auxotrophy for several vitamins, an increased ability to resist bacteriophages and horizontal gene transfer events with the dairy Lactococcus lactis and S. thermophilus as potential donors. In addition, S. macedonicus lacks several pathogenicity-related genes found in S. gallolyticus. For example, S. macedonicus has retained only one (i.e. the pil3) of the three pilus gene clusters which may mediate the binding of S. gallolyticus to the extracellular matrix. Unexpectedly, similar findings were obtained not only for the dairy S. infantarius CJ18, but also for the blood isolate S. pasteurianus ATCC 43144.

Conclusions: Our whole genome analyses suggest traits of adaptation of S. macedonicus to the nutrient-rich dairy environment. During this process the bacterium gained genes presumably important for this new ecological niche. Finally, S. macedonicus carries a reduced number of putative SBSEC virulence factors, which suggests a diminished pathogenic potential.

Figure 1

The circular map of the genome of Streptococcus macedonicus ACA-DC 198. Genomic features appearing from the periphery to the centre of the map: 1. Forward CDSs (red); 2. Reverse CDSs (blue); 3. Putative pseudogenes (cyan); 4. rRNA genes (orange); 5. tRNA genes (green); 6.% GC plot; 7. GC skew.

Figure 2

Chromosome alignments of the Streptococcus bovis/Streptococcus equinus complex members as calculated by progressiveMauve. Chromosome alignments among Streptococcus gallolyticus, Streptococcus macedonicus and Streptococcus pasteurianus(A) and all the aforementioned streptococci and Streptococcus infantarius(B). Local collinear blocks (LCBs) of conserved sequences among the strains are represented by rectangles of the same colour. Connecting lines can be used to visualize synteny or rearrangement. LCBs positioned above or under the chromosome (black line) correspond to the forward and reverse orientation, respectively. The level of conservation is equivalent to the level of vertical colour filling within the LCBs (e.g. white regions are strain-specific). Sequences not placed within an LCB are unique for the particular strain.

Figure 3

Core genome analysis of members of the Streptococcus bovis/Streptococcus equinus complex. Whole CDS Venn diagrams of Streptococcus gallolyticus, Streptococcus macedonicus and Streptococcus pasteurianus(A) or Streptococcus gallolyticus, Streptococcus infantarius, Streptococcus macedonicus and Streptococcus pasteurianus(B). In (B)Streptococcus gallolyticus ATCC 43143 was selected as a representative of the S. gallolyticus species, since it has the longest genome size among the three sequenced strains.