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After a slight hiccup, here is the paper for week 14(ish) of microtwjc!  We all have favourite bugs and this one is fast becoming mine (if its cool enough for history guy Dan Snow and C4 filthy cities episode, its good enough for me )

Title: Poly-N-Acetylglucosamine Expression by Wild-Type Yersinia pestis Is Maximal at Mammalian, Not Flea, Temperatures

Abstract: Numerous bacteria, including Yersinia pestis, express the poly-N-acetylglucosamine (PNAG) surface carbohydrate, a major component of biofilms often associated with a specific appearance of colonies on Congo red agar. Biofilm formation and PNAG synthesis by Ypestis have been reported to be maximal at 21 to 28°C or “flea temperatures,” facilitating the regurgitation of Ypestis into a mammalian host during feeding, but production is diminished at 37°C and thus presumed to be decreased during mammalian infection. Most studies of PNAG expression and biofilm formation by Ypestis have used a low-virulence derivative of strain KIM, designated KIM6+, that lacks the pCD1 virulence plasmid, and an isogenic mutant without the pigmentation locus, which contains the hemin storage genes that encode PNAG biosynthetic proteins. Using confocal microscopy, fluorescence-activated cell sorter analysis and growth on Congo red agar, we confirmed prior findings regarding PNAG production with the KIM6+ strain. However, we found that fully virulent wild-type (WT) strains KIM and CO92 had maximal PNAG expression at 37°C, with lower PNAG production at 28°C both in broth medium and on Congo red agar plates. Notably, the typical dark colony morphology appearing on Congo red agar was maintained at 28°C, indicating that this phenotype is not associated with PNAG expression in WT Ypestis. Extracts of WT sylvatic Ypestis strains from the Russian Federation confirmed the maximal expression of PNAG at 37°C. PNAG production by WT Ypestis is maximal at mammalian and not insect vector temperatures, suggesting that this factor may have a role during mammalian infection.

Importance: Yersinia pestis transitions from low-temperature residence and replication in insect vectors to higher-temperature replication in mammalian hosts. Prior findings based primarily on an avirulent derivative of WT (wild-type) KIM, named KIM6+, showed that biofilm formation associated with synthesis of poly-N-acetylglucosamine (PNAG) is maximal at 21 to 28°C and decreased at 37°C. Biofilm formation was purported to facilitate the transmission of Ypestis from fleas to mammals while having little importance in mammalian infection. Here we found that for WT strains KIM and CO92, maximal PNAG production occurs at 37°C, indicating that temperature regulation of PNAG production in WT Ypestis is not mimicked by strain KIM6+. Additionally, we found that Congo red binding does not always correlate with PNAG production, despite its widespread use as an indicator of biofilm production. Taken together, the findings show that a role for PNAG in WT Ypestis infection should not be disregarded and warrants further study.

This weeks paper can be found here

Usual questions apply…..

  1. Is this paper well written?
  2. Does the data support the conclusion?
  3. What other data/experiments would you like to have seen presented? and most importantly
  4. Where next to determine a role (if there is one?!?) for PNAG in Y. pestis virulence?

Hope everyone can make it to #microtwjc on Tuesday


Sometimes a paper comes along which is simply too good not to include in a journal club. An exciting yet contentious  finding that presents as a paradigm shift in our understanding of a pathogenic microbe with far reaching implications for public health.

Unfortunately I just couldn’t shift my focus to that paper from one essentially about toilets. This weeks paper therefore concerns that which we would rather not know – the diversity, and sources, of surface microbial communities in public restrooms.

The authors present the application of 16S rRNA sequence analysis (apologies, it is another next-gen approach) to further our understanding of microbial communities on surfaces.  They in turn demonstrate the utility of SourceTracker (a Bayesian-based comparison tool) for elucidating the likely source of these contaminations. The implications of their findings to human health are finally discussed.

Some points to consider:

1. Was it really worth it?

2. Was the goal of this study  clear?

3. Given the analysis methods employed can any extrapolation to the prevalence of disease causing bacteria really be made?

As ever please feel free to leave further discussion points……