Distinct genealogies for plasmids and chromosome

An earlier perspective on the diversity of conjugative elements in microbes [1] attempted to provide a broad audience with an introductory overview of the arcane biology of mobile genetic elements and their terminologies. It might well have been entitled "Plasmids, ICEs, IMEs, and Other Mobile Elements for Dummies," but common sense prevailed. This perspective introduces two related articles in the current issue of PLOS Genetics [2,3] and might have equally aptly been entitled "Antibiotic-Resistant Plasmids and Their Epidemiology for Dummies.

Population structures in the SARA and SARB reference collections of Salmonella enterica according to MLST, MLEE and microarray hybridization

In the 1980's and 1990's, population genetic analyses based on Multilocus Enzyme Electrophoresis (MLEE) provided an initial overview of the genetic diversity of multiple bacterial species, including Salmonella enterica. The genetic diversity within S. enterica subspecies enterica according to MLEE is represented by the SARA and SARB reference collections, each consisting of 72 isolates, which have been extensively used for comparative analyses. MLEE has subsequently been replaced by Multilocus Sequence Typing (MLST). Our initial MLST results indicated that some strains within the SARB collection differed from their published descriptions. We therefore performed MLST on four versions of the SARB collection from different sources and one collection of SARA, and found that multiple isolates in SARB and SARA differ in serovar from their original description, and other SARB isolates differed between different sources. Comparisons with a global MLST database allowed a plausible reconstruction of the serovars of the original collection. MLEE, MLST and microarrays were largely concordant at recognizing closely related strains. MLST was particularly effective at recognizing discrete population genetic groupings while the two other methods provided hints of higher order relationships. However, quantitative pair-wise phylogenetic distances differed considerably between all three methods. Our results provide a translation dictionary from MLEE to MLST for the extant SARA and SARB collections which can facilitate genomic comparisons based on archival insights from MLEE

How old are bacterial pathogens?

Only few molecular studies have addressed the age of bacterial pathogens that infected humans before the beginnings of medical bacteriology, but these have provided dramatic insights. The global genetic diversity of Helicobacter pylori, which infects human stomachs, parallels that of its human host. The time to the Most Recent Common Ancestor (tMRCA) of these bacteria approximates that of anatomically modern humans, i.e. at least 100,000 years, after calibrating the evolutionary divergence within H. pylori against major ancient human migrations. Similarly, genomic reconstructions of Mycobacterium tuberculosis, the cause of tuberculosis, from ancient skeletons in South America and mummies in Hungary support estimates of <6,000 years for the tMRCA of M. tuberculosis. Finally, modern global patterns of genetic diversity and ancient DNA studies indicate that during the last 5,000 years plague caused by Yersinia pestis has spread globally on multiple occasions from China and Central Asia. Such tMRCA estimates provide only lower bounds on the ages of bacterial pathogens, and additional studies are needed for realistic upper bounds on how long humans and animals have suffered from bacterial diseases

The B cell response to Plasmodium chabaudi chabaudi malaria in the mouse model

This thesis characterises the B cell response of C57BL/6 mice after infection with Plasmodium chabaudi chabaudi (AS) with regard to plasma cell longevity, B cell memory and the specific response to part of the merozoite surface protein-1 (MSP-1 21), an antigen involved in protection against malaria. Infected spleens showed massive but reversible disruption of the white pulp architecture around the peak of infection. Nevertheless, strong plasma cell and germinal centre responses were detected along with atypical plasma cell location and apparent involvement of marginal zone B cells. The MSP-1 21-specific B cell response to primary infection did not display any obvious abnormalities at the cellular level. The longevity of protection was assessed in the presence and absence of parasites. P. c. chabaudi parasites persisted for 2-3 months after infection. A single infection mediated protection in form of reduced parasitaemias for up to 9 months. When mice were reinfected after 2.5 months but not later, the presence of parasites from the primary infection led to an additional reduction. This can be attributed to cooperation between effector and memory cells. MSP-1 21-specific IgG levels were measured in primary and secondary infections and during the intervening period. Plasma levels of IgG against MSP-1 21 and crude malarial extract drop significantly between 1 and 2. 5 months post infection and then remain constant, suggesting that Plasmodium-specific plasma IgG levels are maintained by long-lived plasma cells independent of parasite presence or absence after day 30. However, MSP-1 21-specific memory B cell generation, maintenance or reactivation appears to be reduced in the presence of parasites albeit with increased affinity maturation. Primary and secondary antibody responses to MSP-1 21 develop more slowly than antibody responses observed in other infections, suggesting that despite the strong B cell response during primary infection with malaria, there is an impairment of the B cell response at some level

Metagenomics of the modern and historical human oral microbiome with phylogenetic studies on Streptococcus mutans and Streptococcus sobrinus

We have recently developed bioinformatic tools to accurately assign metagenomic sequence reads to microbial taxa: SPARSE [1] for probabilistic, taxonomic classification of sequence reads, EToKi [2] for assembling and polishing genomes from short read sequences, and GrapeTree [3], a graphic visualizer of genetic distances between large numbers of genomes. Together, these methods support comparative analyses of genomes from ancient skeletons and modern humans [2,4]. Here we illustrate these capabilities with 784 samples from historical dental calculus, modern saliva and modern dental plaque. The analyses revealed 1591 microbial species within the oral microbiome. We anticipated that the oral complexes of Socransky et al. [5] would predominate among taxa whose frequencies differed by source. However, although some species discriminated between sources, we could not confirm the existence of the complexes. The results also illustrate further functionality of our pipelines with two species that are associated with dental caries, Streptococcus mutans and Streptococcus sobrinus. They were rare in historical dental calculus but common in modern plaque, and even more common in saliva. Reconstructed draft genomes of these two species from metagenomic samples in which they were abundant were combined with modern public genomes to provide a detailed overview of their core genomic diversity

Formal comment to Pettengill : the time to most recent common ancestor does not (usually) approximate the date of divergence

In 2013 Zhou et al. concluded that Salmonella enterica serovar Agona represents a genetically monomorphic lineage of recent ancestry, whose most recent common ancestor existed in 1932, or earlier. The Abstract stated ‘Agona consists of three lineages with minimal mutational diversity: only 846 single nucleotide polymorphisms (SNPs) have accumulated in the non-repetitive, core genome since Agona evolved in 1932 and subsequently underwent a major population expansion in the 1960s.’ These conclusions have now been criticized by Pettengill, who claims that the evolutionary models used to date Agona may not have been appropriate, the dating estimates were inaccurate, and the age of emergence of Agona should have been qualified by an upper limit reflecting the date of its divergence from an outgroup, serovar Soerenga. We dispute these claims. Firstly, Pettengill’s analysis of Agona is not justifiable on technical grounds. Secondly, an upper limit for divergence from an outgroup would only be meaningful if the outgroup were closely related to Agona, but close relatives of Agona are yet to be identified. Thirdly, it is not possible to reliably date the time of divergence between Agona and Soerenga. We conclude that Pettengill’s criticism is comparable to a tempest in a teapot

Accurate reconstruction of bacterial pan- and core genomes with PEPPAN

Bacterial genomes can contain traces of a complex evolutionary history, including extensive homologous recombination, gene loss, gene duplications and horizontal gene transfer. In order to reconstruct the phylogenetic and population history of a set of multiple bacteria, it is necessary to examine their pangenome, the composite of all the genes in the set. Here we introduce PEPPAN, a novel pipeline that can reliably construct pangenomes from thousands of genetically diverse bacterial genomes that represent the diversity of an entire genus. PEPPAN outperforms existing pangenome methods by providing consistent gene and pseudogene annotations extended by similarity-based gene predictions, and identifying and excluding paralogs by combining tree- and synteny-based approaches. The PEPPAN package additionally includes PEPPAN_parser, which implements additional downstream analyses including the calculation of trees based on accessory gene content or allelic differences between core genes. In order to test the accuracy of PEPPAN, we implemented SimPan, a novel pipeline for simulating the evolution of bacterial pangenomes. We compared the accuracy and speed of PEPPAN with four state-of-the-art pangenome pipelines using both empirical and simulated datasets. PEPPAN was more accurate and more specific than any of the other pipelines and was almost as fast as any of them. As a case study, we used PEPPAN to construct a pangenome of ~40,000 genes from 3052 representative genomes spanning at least 80 species of Streptococcus. The resulting gene and allelic trees provide an unprecedented overview of the genomic diversity of the entire Streptococcus genus

Neutral genomic microevolution of a recently emerged pathogen, salmonella enterica serovar agona

Salmonella enterica serovar Agona has caused multiple food-borne outbreaks of gastroenteritis since it was first isolated in 1952. We analyzed the genomes of 73 isolates from global sources, comparing five distinct outbreaks with sporadic infections as well as food contamination and the environment. Agona consists of three lineages with minimal mutational diversity: only 846 single nucleotide polymorphisms (SNPs) have accumulated in the non-repetitive, core genome since Agona evolved in 1932 and subsequently underwent a major population expansion in the 1960s. Homologous recombination with other serovars of S. enterica imported 42 recombinational tracts (360 kb) in 5/143 nodes within the genealogy, which resulted in 3,164 additional SNPs. In contrast to this paucity of genetic diversity, Agona is highly diverse according to pulsed-field gel electrophoresis (PFGE), which is used to assign isolates to outbreaks. PFGE diversity reflects a highly dynamic accessory genome associated with the gain or loss (indels) of 51 bacteriophages, 10 plasmids, and 6 integrative conjugational elements (ICE/IMEs), but did not correlate uniquely with outbreaks. Unlike the core genome, indels occurred repeatedly in independent nodes (homoplasies), resulting in inaccurate PFGE genealogies. The accessory genome contained only few cargo genes relevant to infection, other than antibiotic resistance. Thus, most of the genetic diversity within this recently emerged pathogen reflects changes in the accessory genome, or is due to recombination, but these changes seemed to reflect neutral processes rather than Darwinian selection. Each outbreak was caused by an independent clade, without universal, outbreak-associated genomic features, and none of the variable genes in the pan-genome seemed to be associated with an ability to cause outbreaks