New metagenomics software available

A new publication in BMC Bioinformatics by Eppley et al. is available describing Strainer. Strainer is a metagenomics analysis software package focused on population genetics, from the Banfield lab. The software is available here.

Their summary of the software:

Background
Metagenomic analyses of microbial communities that are comprehensive enough to provide multiple samples of most loci in the genomes of the dominant organism types will also reveal patterns of genetic variation within natural populations. New bioinformatic tools will enable visualization and comprehensive analysis of this sequence variation and inference of recent evolutionary and ecological processes.

Results
We have developed a software package for analysis and visualization of genetic variation in populations and reconstruction of strain variants from otherwise co-assembled sequences. Sequencing reads can be clustered by matching patterns of single nucleotide polymorphisms to generate predicted gene and protein variant sequences, identify conserved intergenic regulatory sequences, and determine the quantity and distribution of recombination events.

Conclusions
The Strainer software, a first generation metagenomic bioinformatics tool, facilitates comprehension and analysis of heterogeneity intrinsic in natural communities. The program reveals the degree of clustering among closely related sequence variants and provides a rapid means to generate gene and protein sequences for functional, ecological, and evolutionary analyses.

Microbial genomics and Metagenomics workshop

For those interested in microbial genomics or metagenomics see the announcement here:

The U.S. Department of Energy Joint Genome Institute (DOE JGI) is offering a five-day workshop on Microbial Genomics and Metagenomics, in Walnut Creek, California, January 7-11, 2008. The workshop includes two days of intensive seminars and three days of hands-on tutorials. The goal is to provide training in microbial genomic and metagenomic analysis and to demonstrate how the cutting-edge science and technology of DOE JGI can enhance your research. Participation is limited to 40 attendees (graduate students, postdocs, and faculty or staff scientists). To register for the workshop, submit your application online from the registration page: http://www.jgi.doe.gov/meetings/mgm/


Note the Workshop is FREE. Yes that is right. FREE.

GME 2007 – Getting feedback on PLoS ONe

While at the GME meeting – I have now been approached by 3 people saying they really liked the discussion I initiated on PLoS One for a paper on metagenomics a few weeks ago. They were not saying they liked my comments but that there was an active discussion about some important topics. I think the function on PLoS One has great potential to engage the broad scientific community in discussions that might have previously been limited to journal clubs. So it is nice to see (1) that people are reading stuff on PLoS One and (2) that they seem to like the commenting function.

PLoS One Commentary Expansion and Metagenomics of Warm Waters

ResearchBlogging.org

People interested in metagenomics and/or marine microbiology should check out a recent paper in PLoS One on Metagenomics of the Deep Mediterranean, a Warm Bathypelagic Habitat.

In this paper, the authors use metagenomic sequencing to study microbes from 3000m depth int he Mediterranean. The most interesting thing about this paper is that the water at this depth is much warmer than water from similar depths that have been previously studied by metagenomics. The authors do some comparative metagenomic analysis and conclude that temperature of the water is more important than depth in determining the suite of organisms and genes that are present in the water. This is a preliminary result but does being to show the potential power of comparative metagenomic analyses.

The other thing people might want to check out is the Commentary on this article. I have been trying to get people in my lab to layer their comments and annotations onto this article as a test of the new PLoS One system. So far a few people in the lab have added comments as have I and more are coming. Others should add comments too. This commenting system is one of the nice new features of PLoS One in my mind, although it is still in beta testing. One thing I am still trying to work out is the trackback system …

Martín-Cuadrado, A., López-García, P., Alba, J., Moreira, D., Monticelli, L., Strittmatter, A., Gottschalk, G., & Rodríguez-Valera, F. (2007). Metagenomics of the Deep Mediterranean, a Warm Bathypelagic Habitat PLoS ONE, 2 (9) DOI: 10.1371/journal.pone.0000914

RNA based metagenomics – bee CCD study suggests a way to survey all microbes at once

After being reprimanded justifiably by one of the authors of the recent paper on bee colony collapse disorder (for a blog I posted and then removed) I have come to realize they used a very interesting approach to metagenomics that I have not seen used extensively before.

Their paper can be found here.

Here is the problem they were faced with – how to survey an sample for ALL the microbes present including both viruses and cellular microbes. The challenge to this is that some viruses have RNA genomes and thus if one simply extracts DNA and sequences it one will not sample any of the RNA viruses. One approach to such a challenge would be to isolate RNA and make cDNA and sequence to sample the RNA viruses. And then to separately isolate DNA and then sequence it either directly (using shotgun sequencing) or indirectly by first amplifying genes with PCR.

They chose a different approach – to isolate RNA and make cDNA and then sequence it. In doing this they in fact do get a sample of both RNA viruses AND cellular organisms. For cellular organisms, since most of the RNA in a cell is ribosomal RNA they get a sample of that organisms rRNA which can be used to say what type of organisms are present. Thus in one fell swoop they in essence sample ALL the microbes present in a sample. I had blogged about this and criticized them because they did not explain in the paper or in the press releases all of this logic, but one of the authors set me straight so I deleted my blog. Then I started thinking about it and realized that this seemed to be a relatively novel approach to metagenomics.

Now – I am not sure if this method is quantitative or exactly how robust it is, but it does provide an alternative to rRNA PCR (which has all the biases of PCR) and also provides an alternative to separately sequencing RNA and DNA from a sample. Certainly many have used RNA to cDNA and then sequencing to survey RNA viruses before. But usually they do this in material in which the cellular organisms have been first removed so that one does not get overwhelmed by the RNA from those organisms. But here, they used the power of massively high throughput sequencing and turned this “problem” of getting RNA from cellular organisms on its head and used it to sample RNA viruses and cellular organisms at the same time.

I do not know if this has been done before — maybe other out there know of examples. But whether or not it has been done before, it is an important approach that should be considered in metagenomic surveys and it also suggests that ANYONE doing metagenomic surveys of microbes might want to purify RNA and save it form samples even if one is going to first focus on DNA.

Metagenomics 2007 presentations available

For those interested in metagenomics, the Metagenomics 2007 meeting (also see Konrad’s blog) has posted video and pdf’s of most of the presentations. You can get everything at the CAMERA web site here. I have posted links and titles below:

Larry Smarr [video] [PDF]

Masahira Hattori, Tokyo University/RIKEN
Length: 56:30 [video] [PDF]

Paul Gilna, UCSD
Length: 5:55 [video]

Jed Fuhrman, University of Southern California
Length: 1:04:01 [video] [PDF]

Dawn Field, Oxford Centre for Ecology and Hydrology

Length: 27:05 [video] <!–[video]–> [PDF]

Janet Jansson, Swedish Univ of Agricultural Sciences
[PDF]

James M. Tiedje, Michigan State University
Length: 27:09 [video] [PDF]

Breakouts:

Medical Metagenomics
Human Biology
Chair: Trevor G. Marshall, Autoimmunity Research Foundation
Speaker: Peter J. Turnbaough, Washington University
Length: 49:32 [video] [PDF]

Computational Metagenomics
Ontology and Standardization
Chair: Jonathan Eisen UC Davis
Length: 44:18 [video]

Discovering opportunity for Bioenergy
Chair: Phil Hugenholtz, JGI
Speaker: Yuri Gorby, JCVI

Length: 50:45 [video] [PDF]

Modeling the natural world
Chair: Trina McMahon, Univ. of Wisconsin-Madison
Speaker: Patrick Schloss, UMass Amherst

Length: 1:00:36 [video] [PDF]

Evolution and population
Chair: Francisco Rodriguez-Valera, Universidad Miguel Hernández in Alicante

Length: 1:00:36 [video]

John Wooley, UCSD (Moderator); various speakers
Length: 40:47 [video]

Metagenomic analyses of corals
Forest Rohwer, San Diego State University
Length: 21:15 [video] [PDF]

Population structure of microbial communities in the world’s oceans
Mitchell Sogin, Marine Biological Lab, Woods Hole
Length: 41:20 [video] [PDF]

Structural metagenomics: Lessons from the first experimentally characterized GOS proteins
Adam Godzik, Burnham Institute
Length: 46:35 [video]

Detailed view of the architecture and implementation of a metagenomics server
Philip Papadopoulos, UC San Diego/SDSC
Length: 24:27 [video] [PDF

Metagenome sequence data management and analysis
Victor M. Markowitz, Lawrence Berkeley National Lab
Length: 34:25 [video] [PDF]

Developing a software workbench for marine ecological genomics
Frank Oliver Gloeckner, Max Planck Institute for Marine Microbiology
Length: 32:39 [video] [PDF]

The Metagenomics RAST server: Automated analysis of Sanger and 454-type metagenomes
Folker Meyer, Argonne National Laboratory
Length: 17:00 [video] [PDF]

Technologies for metagenomics selection and sequencing
Gautam Dantas, Harvard University
Length: 33:30 [video] [PDF]

Micro-Mar: A database for dynamic representation of marine microbial diversity
Ravindra Pushker, University College Dublin
Length: 32:00 [video]

Conclusion
Kayo Arima, UC San Diego
Length: 2:16 [video]

Single Cell Genomics Workshop at Bigelow Laboratory

Just got this email from Ramunas Stepanauskas regarding a workshop on single cell genomics. I had mentioned in my blog last week about how this seems to be a critical technology for the future of environmental microbiology. And if you want to get in early into this new technology — apply to this workshop.

I want to draw your attention to the workshop “Single Cell Alternatives to Metagenomics in Environmental Microbiology”, which will take place in Boothbay Harbor, Maine, during September 9-11:

We want to gather the different groups developing microbial single cell genomics (SCG) methodology, exchange information, and thereby enable the field to make faster progress; to examine the dominant science questions that are best addressed by this powerful new tool. The use of SCG will likely have a major impact on the fields of microbial ecology, evolution, and bioprospecting, by enabling partial or complete genome assembly of the uncultured taxa from complex communities, thus providing a critical link between isolate genomics and metagenomics. We envision workshop participants as a mix of about 30 principal investigators, graduate students, and postdocs.

Please see the website (above) for more information and how to apply.

Sincerely,
Ramunas Stepanauskas

Metagenomics 2007 Update

Some brief notes on the Metagenomics 2007 meeting. For those who do not know – metagenomics is the simultaneous sequencing of the genomes of communities of microbes.

Day 1 : I missed the introductory talk by Larry Smarr (my flight was late). Then there was an opening poster session – a diverse collection of things. My favorite were those involving methods to sequence the genomes of single cells (e.g., one by Ramunas Stepanauskas). Such methods will be critical for getting reference genomes from organisms that are not abundant in a community but might nevertheless be important.

Then there was a dinner and a post dinner talk by Masahira Hattori. The talk was not overwhelmingly interesting … sort of a good example of metagenomics as a fad. Hattori has extensive experience in genome sequencing – having been involved in sequencing the human genome as well as the genomes of other large charismatic furry organisms. As with many people who worked on the human genome, he moved into microbial genomic studies. He has done some interesting things in this arena, the most interesting to me being the sequencing of the genome of a symbiotic bacteria called Carsonella. He did talk about this and I learned something I missed in the paper – this genome was sequenced with the aid of genome amplification methods (see above).

Then Hattori talked about his recent work on human microbiome metagenomics. He has done a lot of work in this area including a big project in which 80,000+ sequence reads were generated for 13 healthy Japanese individuals (both adults and children). I confess much of his results/analyses were not convincing – they seemed to me to be something of the order of “I know I should be doing metagenomics – here is some.” His talk did lead to one of the funnier moments of the meeting – someone asked a question that went something like this

“I know when I go to Japan my intestinal tract changes” leading to a groan from the audience and at least one “A little too much information” comment. The questioner was clearly going somewhere with then but before he got too far, Hattori interjected “It gets much better”

Day 2: Some notes.

Day 2 was a pleasant surprise. For many metagenomics related meetings you hear the same talks and the same data over and over and over again. That was not the case here.

The first talk was by Jed Furman. This was truly inspiring to me. He presented a tour de force of environmental microbiology where metagenomics was simply a component and he interwove the history of metagenomics too. Among his topics – use of ARISA in surveying microbes, studies of the functions present in marine archaea, species richness versus latitude, the scalar nature of sampling for microbes, changes in ocean communities over time, and functional redundancy. This to me is the ideal use of metagenomics – as a tool in other studies. In this sense I disagree with those who say metagenomics is a field. I view it more as a tool. A powerful tool, as Furman showed. But most powerful if used in the context of other studies. In particular he said “Remarkably few experiments other than sequencing have been done in environmental genomics and I hope that will change”

Other talks were by Dawn Field, David Relman, Jim Tiedje, Janet Jansson, Forest Rohwer, and Mitch Sogin. Just a few tidbits from these:

  • Janet Jannson who talked in part about a twin study of microbes associated with Crohn’s disease lauded the idea of a Genomic Encyclopedia for Bacteria and Archaea (a project I am leading at the JGI … if you want to know more stay tuned or ask). Also she referred to humans as “fermenters of microbes” a phrase I kind of like.
  • David Relman gave a good overview of his and others’ work on the human microbiome. One thing he emphasized was the need to remember that rare organisms can be important and that most metagenomics projects do not sample the rare bugs well. And he too then talked about single-cell genomics. He also mentioned a paper I have been waiting for and did not know it had come out. This is a paper on microbial colonization of the human gut in infants by Chana Palmer et al. (with Pat Brown as senior author). In this paper they use a rRNA chip to track microbes colonization infants. This paper is spectacular. Really.
  • Forest Rohwer, who talked most about coral associated microbes and coral damage by people started off by saying “This is my humans suck talk”

There were also breakout sessions, one of which I chaired on “Computational Metagenomics.” Our general conclusion from this – metagenomic informatics is really difficult, we need more interdisciplinary efforts to develop new approaches, and that more money is needed for computational efforts if we want to get the most out of metagenomic data (like we were going to say we need less money). All jokes aside, I do agree with this point – metagenomic sequence analysis is orders of magnitude more complex than analyzing any single-organisms genome. And we need a major ratheting up of computational efforts if we are to get the most out of these data sets that are being produced.

Metagenomics 2007 – streaming live

If you are interested in Metagenomics – log on to the Webcast here.

More info on the meeting can be found here.

Metagenomics Meeting July 11-13, 2007 UCSD

Fro those interested in metagenomics, a meeting will be held July 11-13 at UCSD with many of the major players in the field participating in some way (I am on the planning committee, so no bias here).

This meeting is connected to the new CAMERA Metagenomics Database run out of UCSD (as a Joint Venture with the Venter Institute). I am about to head down to the Scientific Advisory Board meeting for CAMERA. So I am asking anyone out there who is interested in metagenomics to check out CAMERA and let me know what you think. Right now I am I guess kind of an insider/outsider in CAMERA. I am supposed to have a subcontract to Davis to help integrate some phylogenetic tools into the database but alas the Davis office has been a bit lax shall we say about getting my subcontract set up. So until that is set up I guess I am a CAMERA advisor.

Anyway, if anyone has any useful opinions out there about CAMERA I will be happy to share them with those at the SAB meeting.