The fall semester has begun, and with that we have started a new round of recordings of the Microbiology Lab Pod. Our fourth episode was recorded on August 20, and the now-familiar crew (Johan Bengtsson-Palme, Emil Burman, Haveela Kunche and Anna Abramova) has been augmented with two new master students in the lab: Sebastian Wettersten and Mahbuba Lubna Akter. This time, we discuss microbial communities of dead and alive deep-sea hydrothermal vents, look at a model system for pathogenic biofilm formation in the lungs, and check in on why patients with covid-19 commonly lose their sense of smell.
The specific papers discussed in the pod (with approximate timings) are as follows:
- 11:30 – Hou, J., Sievert, S.M., Wang, Y. et al., 2020. Microbial succession during the transition from active to inactive stages of deep-sea hydrothermal vent sulfide chimneys. Microbiome 8, 102. https://doi.org/10.1186/s40168-020-00851-8
- 28:45 – Harrington, N.E., Sweeney, E., Harrison, F., 2020. Building a better biofilm – Formation of in vivo-like biofilm structures by Pseudomonas aeruginosa in a porcine model of cystic fibrosis lung infection. Biofilm 2, 100024. https://doi.org/10.1016/j.bioflm.2020.100024
- 52:30 – Brann, D.H., Tsukahara, T., Weinreb, C., et al., 2020. Non-neuronal expression of SARS-CoV-2 entry genes in the olfactory system suggests mechanisms underlying COVID-19-associated anosmia. Science Advances 6, eabc5801. https://doi.org/10.1126/sciadv.abc5801
- 71:45 – Chen, M., Shen, W., Rowan, N.R., et al., 2020. Elevated ACE2 expression in the olfactory neuroepithelium: implications for anosmia and upper respiratory SARS-CoV-2 entry and replication. European Respiratory Journal 2001948. https://doi.org/10.1183/13993003.01948-2020
- 77:15 – Zhang, X., Wang, J., 2020. Deducing the Dose-response Relation for Coronaviruses from COVID-19, SARS and MERS Meta-analysis Results. medRxiv. https://doi.org/10.1101/2020.06.26.20140624
- 78:30 – Sekine, T., Perez-Potti, A., Rivera-Ballesteros, O., et al., 2020. Robust T cell immunity in convalescent individuals with asymptomatic or mild COVID-19. Cell. https://doi.org/10.1016/j.cell.2020.08.017
- 79:45 – Mateus, J., Grifoni, A., Tarke, A., et al., 2020. Selective and cross-reactive SARS-CoV-2 T cell epitopes in unexposed humans. Science eabd3871. https://doi.org/10.1126/science.abd3871
- 80:30 – Lv, H., Wu, N.C., Tsang, O.T.-Y., et al., 2020. Cross-reactive Antibody Response between SARS-CoV-2 and SARS-CoV Infections. Cell Reports 31, 107725. https://doi.org/10.1016/j.celrep.2020.107725
The podcast was recorded on August 20, 2020. If you want to reach out to us with comments, suggestions or other feedback, please send an e-mail to podcast at microbiology dot se or contact @bengtssonpalme via Twitter. The music that can be heard on the pod is composed by Johan Bengtsson-Palme and is taken from the album Cafe Phonocratique.
One thing that I find slightly annoying is when people do not get the basic concepts right – or when debatable concepts are used without discussion of their implications. This further annoys me when it is done by senior scientists, who should know better. Sometimes, I guess this happens out of ignorance, and sometimes to be able to stick your subject to a certain buzzword concept. Neither is good, even though the former reason is little more forgivable then the latter. One area where this problem becomes agonizingly evident is when molecular biologists or medical scientists moves into ecology, as has happened with the advent of metagenomics. When the study of the human gut microflora turned into a large-scale sequencing effort, people who had previously studied bacteria grown on plates started facing a world of community ecology. However, I get the impression that way too often these people do not ask ecologists for advice, or even read up on the ecological literature. Which, I suppose, is the reason why medical scientists can talk about how the human gut microflora can “evolve” into a stable community a couple years after birth, even though words such as “development” or “succession” would be much more accurate to describe this change.
The marker gene flaw
To set what I mean straight, let us compare the human gut to a forest. If an open field is left to itself, larger plants will slowly inhabit it, and gradually different species will replace each other, until we have a fully developed forest. Similarly, the human gut microflora is at birth rather unstable, but stabilizes relatively quickly and within a few years we have a microbial community with “adult-like” characteristics. To arrive at this conclusion, scientists generally use the 16S (small sub-unit) genetic marker to study the bacterial species diversity. This works in pretty much the same way as going out into the forest and count trees of different kinds.
Now, if I went out into the forest once and counted the tree species, waited for 50 years and then did the same thing again, I would presumably see that the forest species composition had changed. However, if I called this “evolution”, fellow scientists would laugh at me. Raspberry bushes do not evolve into birches, and birches do not evolve into firs. Instead, ecologists talk about “succession”; a progressive transformation of a community, going on until a stable community is formed. The concept of succession seems well-suited also to describe what is happening in the human gut, and should of course also be used in that setting. The most likely driver of the functional community changes is not that some bacterial species have evolved new functions, but rather that bacterial species performing these new functions have outcompeted the once previously present.
In fact, I would argue that it is impossible to study evolution through a genetic marker such as the 16S gene (except in the rare case when you study evolution of the 16S gene itself). Instead, the only thing we could assess using a marker gene is how the copy number of the different gene variants change over time (or space, or conditions). The copy number tells us about the species composition of the community at a given time, which can be used to measure successional changes. However, evolutionary changes would require heritable changes in the characteristics of biological populations, i.e. that their genetic material change in some way. Unless that change happens in the marker gene of choice, we cannot measure it, and the alterations of composition we measure will only reflect differences in species abundances. These differences might have arisen from genetic (i.e. evolutionary) changes, but we cannot assess that.
What are we studying with metagenomics?
This brings us to the next problem, which is not only a problem of semantics and me getting annoyed, but a problem with real implications. What are we really studying using metagenomics? When we apply an environmental sequencing approach to a microbial community, we get a snapshot of the genetic material at a given time and site; at specific conditions. Usually, we aim to characterize the community from a taxonomic or functional perspective, and we often have some other community which we want to compare to. However, if we only collect data from different communities at one time point, or if we only study a community before and after exposure, we have no way of telling if differences stem from selective pressures or from more a random succession progress. As most microbial habitats are not as well studied as the human gut, we know little about microbial community assembly and succession.
Also, in ecology a disturbance to a particular community is generally considered as a starting point for a new succession process. This process may, or may not, return the community to the same stable state. However, if the disturbance was of permanent nature, the new community will have to adapt to the new conditions, and the stable state will likely not have the same species distribution. Such an adaption could be caused by genetic changes (which would clearly be an evolutionary process), or by simple replacement of sensitive species with tolerant ones. The latter would be a selective process, but not necessarily an evolutionary one. If the selection does not alter the genetic material within the species, but only the species composition, I would argue that this is also a case of succession.
Complications with resistance
This complicates the work with metagenomic data. If we study antibiotic resistance genes, and say that bacteria in an environment have evolved antibiotic resistance, we base that assertion on that genes responsible for resistance have either evolved within the present bacteria, or have (more likely) been transferred into the genomes of the bacteria via horizontal gene transfer. However, if the resistance profile we see is simply caused by a replacement of sensitive species with resistant ones, we have not really discovered something new evolving, but are only witnessing spread of already resistant bacteria. In the gut, this would be a problem by itself, but say that we do the same study in the open environment. We already know that environmental bacteria have contained resistance genes for ages, so the real threat to human health here would be a spread from naturally resistant bacteria to human pathogens. However, as mentioned earlier, without extremely well thought-through methodology we cannot really see such transmissions of resistance genes. Here, the search for mobile elements, and large-scale takes on community composition vs. resistance profiles in contaminated and non-polluted areas can play a huge role in shedding light on the question of spreading. However, this will require larger and better planned experiments using metagenomics than what is generally performed at the moment. The questions of microbial community assembly, dispersal, succession and adaption are still largely unanswered, and our metagenomic and environmental sequencing approaches have just started to tinker around with the lid of the jar.