Microbiology, Metagenomics and Bioinformatics

Johan Bengtsson-Palme, University of Gothenburg | Wisconsin Institute for Discovery

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Let me get straight to something somewhat besides the point here: summer students can achieve amazing things! One such student I had the pleasure to work with this summer is Shruthi Magesh, and a preprint based on work she did with me at the Wisconsin Institute for Discovery this summer just got published on bioRxiv (1). The preprint describes a software tool called Mumame, which uses database information on mutations in DNA or protein sequences to search metagenomic datasets and quantifies the relative proportion of resistance mutations over wild type sequences.

In the preprint (1), we first of all show that Mumame works on amplicon data where we already knew the true outcome (2). Second, we show that we can detect differences in mutation frequencies in controlled experiments (2,3). Lastly, we use the tool to gain some further information about resistance patterns in sediments from polluted environments in India (4,5). Together these analyses show that one of the most central aspects for Mumame to be able to find mutations is having a very high number of sequenced reads in all libraries (preferably more than 50 million per library), because these mutations are generally rare – even in polluted environments and microcosms exposed to antibiotics. We expect Mumame to be a useful addition to metagenomic studies of e.g. antibiotic resistance, and to increase the detail by which metagenomes can be screened for phenotypically important differences.

While I did write the code for the software (with a lot of input from Viktor Jonsson, who also is a coauthor on the preprint, on the statistical analysis), Shruthi did the software testing and evaluations, and the paper would not have been possible hadn’t she wanted a bioinformatic summer project related to metagenomics, aside from her laboratory work. The resulting preprint is available from bioRxiv and the Mumame software is freely available from this site.

References

  1. Magesh S, Jonsson V, Bengtsson-Palme JQuantifying point-mutations in metagenomic data. bioRxiv, 438572 (2018). doi: 10.1101/438572 [Link]
  2. Kraupner N, Ebmeyer S, Bengtsson-Palme J, Fick J, Kristiansson E, Flach C-F, Larsson DGJ: Selective concentration for ciprofloxacin in Escherichia coli grown in complex aquatic bacterial biofilms. Environment International, 116, 255–268 (2018). doi: 10.1016/j.envint.2018.04.029 [Paper link]
  3. Lundström S, Östman M, Bengtsson-Palme J, Rutgersson C, Thoudal M, Sircar T, Blanck H, Eriksson KM, Tysklind M, Flach C-F, Larsson DGJ: Minimal selective concentrations of tetracycline in complex aquatic bacterial biofilms. Science of the Total Environment, 553, 587–595 (2016). doi: 10.1016/j.scitotenv.2016.02.103 [Paper link]
  4. Bengtsson-Palme J, Boulund F, Fick J, Kristiansson E, Larsson DGJ: Shotgun metagenomics reveals a wide array of antibiotic resistance genes and mobile elements in a polluted lake in India. Frontiers in Microbiology, 5, 648 (2014). doi: 10.3389/fmicb.2014.00648 [Paper link]
  5. Kristiansson E, Fick J, Janzon A, Grabic R, Rutgersson C, Weijdegård B, Söderström H, Larsson DGJ: Pyrosequencing of antibiotic-contaminated river sediments reveals high levels of resistance and gene transfer elements. PLoS ONE, Volume 6, e17038 (2011). doi:10.1371/journal.pone.0017038.

Can you spot the differences in color shade in this picture?

Me neither. And that’s why I suspect that I’ve spent 30 hours or so on something completely not useful stuff. I hereby take weekend. See you next week!

I’ve been having a very intense start of the year with the move to the US and getting the family accustomed to Madison (which has taken time and energy, but gone really well). I just wanted to make you aware of that I have started posting at the Wisconsin Blog again and hope to be sharing research related stuff from my year in the US there. For more personal stuff, our family has set up a blog (in Swedish) at this address: https://palmeiamerikat.blogspot.com. You are very welcome to follow our adventure there!

It’s been a long time before I have written something here, mostly because making ourselves at home in Madison have take some time; then we go the flue; and then there have been a lot at work after that. But now i will try to have another go at writing somethings on the Wisconsin Blog. First, a look at my (already messy) desk here at the Wisconsin Institute for Discovery.

And then, a look at my lab space, which I have a view of straight from my desk, through a glass window.

This week, I have started experiments with exposing our little model community to antibiotics and it looks like I’m getting potentially exciting results. I have to sit down with the data today to see if there’s statistical differences, but from the looks of the biofilms, there is potential here.

Next week I will try to start experiment with sand columns and see if I can replicate some of this in this setting as well. It is interesting being back in the lab, and I feel that this an experience that will be very valuable for me going forward. I look forward to the days later this spring when I will start generating sequence data from my own experiments!

Leaving Madison

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Jorn's Hall

Jorn's Hall - will not be missed

So this morning, I packed my stuff, cleaned out my room and went to WID for the last time this year. I had time to say goodbye to everyone in the lab but Bailey, and I have to admit that I feel a little bit sad leaving. This is a really good place, filled with very good people, in a very beautiful town. I will miss you Handelsman lab, and I will miss you Madison. On the other hand, tomorrow I will be picking up my daughter after preschool for the first time in a month.

My plane

Waiting for my flight

I am now waiting for my flight at the airport. I prepare for a long night of lost sleep (as I cannot sleep on airplanes). I am endlessly happy that I will be able to combine these two fantastic worlds next year when we are moving here altogether. Until then, farewell Madison – see you soon again.

Wisconsin Institute for Discovery

Wisconsin Institute for Discovery - a place filled with people I will miss

I guess this concludes the blog for this time around. Please check back this winter when we’re going here the next time. This is Johan in Madison – over and out.

Lake Mendota

Lake Mendota

Conjugation and Transduction

Copper to treat Anemia

Copper to treat Anemia

Knockout mice

Knockout mice

Vitamin D derivatives

Vitamin D derivatives

Around the biochemistry building of the University of Wisconsin-Madison campus (and perhaps also elsewhere), there are a lot of these signs, highlighting discoveries that were made in Madison. And they really have a history to be proud of. This place is where both bacterial conjugation and transduction was discovered, paving the way for much of the genome editing and studies we take for granted today. And they also did the basics required to generate knock-out mice for genetics studies. I could go on, but I I think I made my point. This place has a lost of history. And I doubt that my slightly messy project will live up to these groundbreaking discoveries.

Labor day

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My desk at WID

The place I work at WID... for two more days

This Monday is Labor day in the US. Which means, I work. After having Wednesday off from lab work, me and Manuel finished up my final experiments for this round yesterday evening, so I have spent this afternoon doing some data analysis. Or rather, I should have spent the day doing data analysis, but it was boring, so instead I have mostly been scripting this… Hopefully that will pay off at a later date when I analyze more plates…

Before that, I went to see a house, which I liked a lot, so hopefully I will be able to sign a contract for this before I leave for Sweden again on Wednesday. Fingers crossed, over and out!

Farmers Market

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State Capitol and Farmers Market

State Capitol and Farmers Market

Today, I went to the big Saturday thing in Madison – the Farmers Market. Apparently this is one of the largest (if not the largest) farmers markets in America, and it’s huge. Farmers (and sellers of some other stuff like yarn) gather all around the State Capitol, and when I arrived at around half past eleven, there were so many people there that you had to just enter into the crowd and follow it in the given walking direction. The sign “Stay right for hot & spicy cheese bread” was right on it; you had to stay in the right lane to get there. But there was so much nice stuff. And then on the opposite side of the stands, there were families picnicking on the lawn and kids running around. I imagine that this will be a nice Saturday outing next Spring when we bring the kids here. For now, I ended up with three apples and some sprouts. I need to finish the food in the fridge before I go back home on Wednesday. Is it time to go home already? It’s been an eternity, but time has also passed so fast…

Madison Farmers Market

Madison Farmers Market


Stay right for bread

Stay right for hot & spicy cheese bread

The rest of the day I spent walking around by the Lake Monona and Lake Wingra, taking a break for a coffee at Colectivo Coffee. Very nice place with excellent pourovers. But now my feet hurts and I need to get some sleep!

Vegetables

Vegetables

Tomatos

Tomatos

Aubergines

Aubergines

Red peppers

Red peppers or chilies or whatever they are called

Surface tension on 96-well plate

Surface tension - or what we have been working with over the last few days

I’ve spend the lion’s share of the past few days in the lab, working quite late to obtain supercool results that we will now verify over the weekend. It’s very exciting to be able to generate useful data already during your first week in the lab (after not doing wet lab work for years), but mostly this is thanks to Manuel. Anyway, look at the beauty of those plates! My new favorite color must be crystal violet.

Students are back

Students are back, and with that 'all campus worship', whatever that is

Another thing that has happened over the past week is that the students are beginning to come back her for the autumn semester. This means nice little paintings on the ground, lots of things happening downtown, but also that it’s harder to sleep since the previously quiet area of college dorms is not seeing quite a significant amount of nightlife. Large packs of students passing by my window at night, talking loudly. It’s mostly nice though. There are many parallels between Uppsala and Madison – both are cities heavily centered around their universities, and both seem to die a little during summers when the students move out. The differences mainly are that Madison is much more beautifully located between three lakes, that Uppsala has a much older history, and that Madison seems to have quite a bit of nightlife also in the summertime.

Almost there

Almost there...

On Monday I will go to see a house to rent next year. Please keep your fingers crossed that it’s good and, if so, that we’ll get it! Otherwise housing has been a bit of a hassle, since everyone wants to rent now before the autumn semester. But I hope that that problem will no longer be a problem on Monday. Have a nice weekend!

On my way to work

On my way to work

Okay, first of all this is a shoutout post to Manuel Garavito, who has been putting up with me for the last two days – my first two days in a wet lab for several years. Manuel has been beyond fantastic in showing me their basic protocols and having patience with my rusty lab skills. If this project will work out, it will very much be because of him.

The Hop CatThis weekend, there haven’t been a lot of time for other things than lab work, but I spent Friday evening at the Memorial Union where I happened to stumble into a concert with Brazilian (I think, not sure) music. And on Saturday evening I went out to the Hop Cat, where I tasted the fantastic beer Psychedellic Cat Grass and got taught the basics of American football by a woman who was also there on her own, apparently to watch the game. So, I’m doing my well but working my ass off with things I am not really that good at. Yet.