Tuesday, December 8, 2015

Horizontal gene transfer from two bacterial genomes into one animal nucleus

For the past two weeks, the science of horizontal gene transfer (exchange of DNA between unrelated organisms such as between microbes and animals) has been a source of scholarly debate, specifically for the case of tardigrades (see Atlantic article). Our latest lab publication in PeerJ (an affordable, transparent, and open access journal) by first author @DNAdiva87 will not cause such a ruckus. Based on DNA sequencing, analysis, and importantly staining of the bacterial inserts in the animal genome, we show for the first time transfer of hundreds of genes from two different Wolbachia bacteria into one host grasshopper genome. Graphical and text abstract are enclosed below. It is also, as of today, featured at the top of the PeerJ journal site.

This work could not have been done without the talents of a great scientist like Lisa and our colleagues in Spain. They and the work were featured in a podcast for "This Week in Microbiology" on 12.17.15. You can check it out here from time 40:48 - 54:25.

Hybrid zones and the consequences of hybridization have contributed greatly to our understanding of evolutionary processes. Hybrid zones also provide valuable insight into the dynamics of symbiosis since each subspecies or species brings its unique microbial symbionts, including germline bacteria such as Wolbachia, to the hybrid zone. Here, we investigate a natural hybrid zone of two subspecies of the meadow grasshopper Chorthippus parallelus in the Pyrenees Mountains. We set out to test whether co-infections of B and F Wolbachia in hybrid grasshoppers enabled horizontal transfer of phage WO, similar to the numerous examples of phage WO transfer between A and B Wolbachia co-infections. While we found no evidence for transfer between the divergent co-infections, we discovered horizontal transfer of at least three phage WO haplotypes to the grasshopper genome. Subsequent genome sequencing of uninfected grasshoppers uncovered the first evidence for two discrete Wolbachiasupergroups (B and F) contributing at least 448 kb and 144 kb of DNA, respectively, into the host nuclear genome. Fluorescent in situ hybridization verified the presence of Wolbachia DNA in C. parallelus chromosomes and revealed that some inserts are subspecies-specific while others are present in both subspecies. We discuss our findings in light of symbiont dynamics in an animal hybrid zone.
Cite this as
Funkhouser-Jones LJ, Sehnert SR, Martínez-Rodríguez P, Toribio-Fernández R, Pita M, Bella JL, Bordenstein SR. (2015Wolbachia co-infection in a hybrid zone: discovery of horizontal gene transfers from two Wolbachia supergroups into an animal genomePeerJ 3:e1479 

Sunday, December 6, 2015

Getting the Hologenome Concept Right

Related publication: Getting the hologenome concept right: an eco-evolutionary framework for hosts and their microbiomes (2016)

What is the target of natural selection in animals and plants? From a phenotypic perspective, selection sees trait variation that influences who passes on offspring and who does not. Given this rather obvious tenet, selection at the individual level is effectively selection on the holobiont - the community of microrganisms and its host. Consequently, variation in the collective genomes of the holobiont - i.e, the hologenome - yields variation in phenotypes upon which evolution can act. This hologenome concept of evolution distinguishes itself by placing emphasis on the vast symbiotic complexity and transmission routes inherent in holobiont animals and plants. As we emphasized in a recent perspective, it is also true that just as  large parts of the nuclear genome can evolve neutrally or be in conflict, so too can large parts the hologenome.

Yesterday, I read a skeptical essay on the hologenome concept by Nancy Moran and Daniel Sloan.  I have several thoughts on it. In short, it was great to see the attention on the topic, as all opinions and analyses are welcome at this nascent stage of the framework. Raising the profile of this area was precisely the intent of a recent publication with Kevin Theis, in which we clarified the history and use of the terms, revised the hologenome concept to include new perspectives such as neutrality, and laid a foundation for hypothesis-driven research and theoretical analyses for the field. This article subsequently calls for some of the same, which is great, but it is unnecessarily narrow and divisive at times. I briefly lay out the misconceptions about the article.

From the start in paragraph 2, it wasn't clear if M&S read our publication in detail. They attribute the origin of the word "holobiont" to Mindell (1992) but we showed that it was first used by Lynn Margulis in 1991. I had the pleasure of corresponding with Mindell prior to our publication, and he humbly tipped us off to the false credit. As we state...
The term "holobiont" traces back to Lynn Margulis and refers to symbiotic associations throughout a significant portion of an organism's lifetime, with the prefix holo- derived from the Greek word holos, meaning whole or entire. Amid the flourishing of host microbiome studies, holobiont is now generally used to mean every macrobe and its numerous microbial associates [19,22]
Second, M&S read our article well enough to extract some quotes, which I fully stand by. But they seem to be skeptical while at the same time they do not elaborate on the justifications spelled out in the paper. Take the example of thinking about host-microbe interactions as interspecific epistasis, similar to gene-gene interactions in the same genome. This is just scaling well accepted population genetic epistasis to community genetic epistasis and it has been done by many eco-evolutionary biologists. As we discuss in our essay, theory shows that there is an intellectual continuum between epistasis in the same genome and interspecific epistasis between different genomes, even for symbionts that are horizontally transmitted. This is not controversial or problematic. What is up for experimentation is the degree to which the epistasis comes and goes versus persist for selection to operate on it. The challenges are interestingly the same for both genetic epistasis in the same genome and intergenomic epistasis. There is nothing out of the ordinary here.

Third, I heartily agree with M&S that coevolution has run amok in the microbiome field (without any evidence) and has been applied incorrectly in most cases. Glad to see this getting more attention. It is precisely why I came up with the word "phylosymbiosis" in 2012-2013 because it does not confer coevolution or codiversification apriori to the observed pattern of microbial community relationships recapitulating host phylogenetics.  It does, however, provide evidence for community selection in controlled studies of the microbiota where microbes are assembled in a deterministic manner that parallels host genetic relationships. Controlled studies of Nasonia wasps and Hydra best show this pattern. I also like their Figure 1 on phylosymbiosis and think that it could be improved as both models could be operating at the same time. Some microbes could be strictly evolving in synchrony with hosts by vertical transmission while other microbes may be horizontally transmitted.

Fourth, the essay ignores that hologenomic models fully accommodate mutualism and parasitism. It is a misconception that it does not. Just as selfish genes are part of the nuclear genome, pathogens are part of the hologenome. See Principle 8 of our essay.

  1. VIII. The hologenome is shaped by selection and neutrality
    • Natural selection can work to remove deleterious nuclear mutations or microbes while spreading advantageous nuclear mutations or microbes; in the absence of selection, the neutral spread of hologenomic variation through populations is an inherently stochastic process.
    • Mixed ecological models of stochastic and deterministic community assembly likely reflect natural systems, and partitioning the microbiota into stochastic versus deterministic subunits will be an important future goal of the field.
Fifth, the hologenome concept fits squarely into multi-level selection theory. Most would agree that it is not correct to say that it is the only unit of selection, but it is equally not correct to say that the concept fails as a whole if there are others level of selection operating. Many levels are operating simultaneously. Moreover, for phenotypic selection, the key point is that nearly all individual selection is holobiont-level selection given the pervasive affects of the microbiota on host biology. The genetic response to selection is what's clearly more challenging to unravel and whether this occurs at a holobiont-level or not is indeed an experimental question worthy of future attention that people are working on.

  1. VII. The hologenome concept fits squarely into genetics and accommodates multilevel selection theory
    • Multilevel selection theory asserts that selection operates across multiple levels of genetic variation with phenotypic effects, from genes to hologenomes and beyond.
    • Holobionts are exclusive to hosts and their associated microbiota; different holobionts, such as a pollinator and a flower, interact with each other under standard ecological principles.
Sixth, the paper frequently plays both sides of the debate. Obligate endosymbionts and organelles are presented as hologenomic entities, but then the paper at times claims hologenomic evolution could be relatively insignificant or rare. These statements are contradictory.

The paper's abstract concludes that "Although selection at the level of the symbiotic community, or hologenome, occurs in some cases, it should not be accepted as the null hypothesis for explaining features of host-symbiont associations". This opinion is largely based on the inaccurate pretense that scientists are claiming holobiont-level selection without evidence or that holobionts are a kumbaya conglomerate. Neither is true. It is easy to oversimplify any concept to death if one wants to. Yet as Einstein once noted "Everything should be made as simple as possible but not simpler".

Tuesday, November 24, 2015

Q&A on Holobionts and Hologenomes

Image from Brucker and Bordenstein, 2013, Zoology
In September, a few friends on twitter entered into a lively discussion on holobionts and hologenomes that was subsequently borne out in a series of Q&A emails. Below, we summarize this discourse - whose goal was to flesh out some of the nuances and foundations of the holobiont and hologenome terms (related blog post  - What are holobionts and hologenomes?) and concepts. David Baltrus poses queries to Seth Bordenstein and Kevin Theis. What follows is very informal, but useful for the three of us and perhaps our colleagues. We all hope it is helpful from the various angles that one may look at this stimulated area. 

Q1: The first point that I think still needs clarifying after the hologenome article and the perspective piece in Science is what the point of promoting “the hologenome” actually is. I’m probably just too close...but to me it’s not a controversial idea that microbes matter for evolution/ecology of macro-organisms.

A1 (SB): There’s two ways to answer this question. First, I think that this is an uncertain argument as one could raise that same question for virtually any term. Why do we need the term “metagenome” if we knew there were previously microbial genomes in soil, or why do we need the term "selfish gene” if we already knew there were transposons, etc. We could stop there, but I think these terms clearly have uses now that one would not question. Indeed, the selfish gene debate early on is very similar to the holobiont and hologenome terms; moreover I think it goes deeper than just this. We’re not just using words willy nilly :) There hasn’t been a good singular term yet to describe the entity of the multispecies organism and their genomes. In fact, we argue its been quite the opposite as we describe in principle 2 (link to paper):

There appears to be a considerable number of misplaced characterizations and colloquialisms used to refer to host-microbiota symbioses, and these misnomers can potentially act as impasses to new advances”.

We need to chuck out all the superorganism and organ system colloquialisms for good reasons. Moreover, the holobionts and hologenomes properly emphasize the diverse and complex array of symbionts and their genomes in one host species. In contrast to binary symbioses, the multispecies consortia are relatively new and universal features of macrobes that emerged within the last decade for many biologists. That’s what the terms best capture in a timely fashion. See Gordon, Knowlton, Reman, Rohwer, Youle for similar arguments (link).  

A1 (KRT): Yes, it is readily accepted that microbes matter for the evolution/ecology of macro-organisms, but importantly the hologenome concept goes beyond this observation in a very substantive way. Specifically, it posits that the host and its microbiota are a potential primary unit of selection. It is this component of the concept, along with the emphasis on the diversity of the symbiotic communities to be considered, as SB already noted, that requires the use of new terminology. All macro-organisms are populated by microbes, these associations are often not random but rather are replicated across macro-organismal generations and are reflective of their phylogeny, and these associations also often profoundly affect the phenotypes of what we typically think of as individual macro-organisms. As we note in the paper:

the debatable and testable issue of the hologenome is whether nuclear genes and microbes are coinherited to a degree that evolution can operate on their interaction.”

Q2: In light of my own point #1 above, almost all of the PLoS principles paper is couched in terms of microbial “symbionts”. While I gather that you are using the broad term “symbionts” here which includes pathogens, it seems to me that all the points that the piece brings up can be flipped and looked at from the point of view of pathogens as well. I don’t think any of this is controversial if it’s couched in terms of microbial pathogens, especially long term chronic ones. HIV can be environmentally acquired or vertically inherited and certainly causes changes in human phenotypes/evolution/ecology. These phenotypic changes (i.e. loss of T cells) can appear to “reboot” some aspects of Lamarck’s ideas just as much as beneficial microbes, only in this case the acquired phenotype is disease. Do pathogenic infections count for rebooting Lamarckian principles if they are inherited across generations? Seems like it's the same story, but you guys put the Larmarckian thing in there to be a bit provocative, which is completely understandable, it just kind of sticks out to me. I don’t think that you see a distinction between beneficial and pathogenic microbes, but I think at least some of the ideas mentioned have been pretty thoroughly worked out for pathogens (speciation for one). If anything, I might argue that pathogens have more power to coevolve with humans than beneficial microbes because of stronger selection pressures on both ends (handwaving over a lot of assumptions).

A2 (SB): I'm trying to follow the logic here, as it seems you are playing both sides of the Q/A. First, you are right in that we have always used symbionts in our publications to refer to all types of associations. And we do not subscribe to overselling one phenotype for another. Quite the contrary, we write that

In the microbiome, selection favors the spread of beneficial microbes involved in nutrition, defense, or reproduction, while pathogenic microbes are either purged by holobiont selection or the pathogens deploy adaptations….”

Second, there is the issue of Lamarck. You agree with Lamarckian aspects of pathogens, but then questions if using Lamarck in the paper is too provocative. Mmmm…can’t have it both ways :) Our goal is to get the science right in its most accurate way, not be provocative in a purposeful sense. If that accuracy or view of science happens to be taken as provocative, that might say something more about where the community is now, than our intent per se. Indeed, Margaret McFall-Ngai once told similar stories for some of her perspectives on the immune system’s role in regulating beneficial microbes. The immunologists were initially up in arms. Finally and to your last point, it is the very fact that many aspects of symbioses have been worked out that we can reason the holobiont and hologenome concepts are the best model to explain the complexity of the systems in a comprehensive light. I think Kevin and I both agree also that its’ not perfect. It needs to be made better with more concepts, theory, and experimentation that is the stated intent of the essay.

A2 (KRT):  We do employ “symbiont” broadly, referring to resident microbes: beneficial, pathogenic, or otherwise. The hologenome concept accommodates each of these interaction types. Lamarckian evolution is simply heritability of acquired characteristics. If acquired symbionts have fitness consequences and are thereafter heritable, they are consistent with Lamarckian evolution. I do not see that as provocative. It should, however, be noted that although we present the argument for a Lamarckian element to the hologenome concept, the Rosenbergs (and many others) of course originally did so (link).

In general, strength of selection may be greater on pathogenic interactions as you suggest, but it can be substantial on beneficial interactions as well. The Rosenbergs’ coral example comes readily to mind (link). Regardless, this is not consequential to the validation or refutation of the hologenome concept as again it accommodates all interaction types. Lastly, I concur with SB, the hologenome concept is not perfect, and in the essay we discuss where future efforts should be focused, but it is the most comprehensive evolutionary framework we currently have for discussing and investigating the ecological and evolutionary complexities of host-microbial systems.

Q3: We have a pretty good vocabulary for ideas like co-evolution, multi-level selection, phenotypic plasticity. What makes the hologenome different enough to warrant special mention in light of a lot of previous theory and literature? If it’s combination of all these things, all of these ideas must inherently incorporate each other in nature anyway, no?

A3 (SB): I think A1 mostly applies here too, but let me know if I'm missing something. Most importantly, we haven’t had a word/singular framework that captures the fact that all of these evolutionary and ecological processes are happening at once across the complexity of the holobiont. We’ve traditionally seen host-microbe interactions as interactions between individual host and individual microbe. The holobiont and holgoenome concepts instead place emphasis on the complex systems biology of the whole system driving evolution. For many phenotypes (and as Charles Goodnight initially put in an email to me), so-called "individual selection" will actually be selection on the community, with many players affecting the selection, and that is what we as a community haven’t appreciated enough.

A3 (KRT): Nice explanation SB. I will only add that, for me at least, the holobiont concept is a redefining of that which constitutes an individual animal or plant. This new construct, or unit, requires a name and a clear definition. That definition is itself ‘evolving,’ hopefully catalyzed to some degree by our essay.

Q4a: You say that the hologenome is “not complex” and point out that they are really only referring to microbes (see microbiome in Title).

A4a (SB): Mmmm, you’re taking words out of context here. We actually state the opposite:

The holobiont and hologenome concepts upgrade this conventional vision to encompass the vast ecological and genomic COMPLEXITY of a host and its total microbiota

And the particular quote excised above was from the following paragraph:

For instance, do the genomes of an insect pollinator and flower constitute a hologenome? The ANSWER HERE IS NOT COMPLEX. Holobionts and their hologenomes are exclusive to the hosts and their associated microbiota. Different holobionts, such as the aforementioned pollinator and flower, clearly interact, but these interactions are not new to biology, as they form the basis for all past and present ecological investigations. They are simply holobionts themselves interacting with each other."

Q4b: My read of this has always been that I don’t know why microbes should be viewed any differently than macrobes that have close associations with humans. The presence of worms can affect distributions of gut microbes across animal life. Macrobes can have just as much an effect on human evolution as microbes. Why the distinction? Is there a distinction? Is it because that humans/etc...are hosts for microbes? Humans are hosts for worms and mites over the course of their whole lives too. Worms and mites can show patterns of vertical inheritance just like microbes. Why are worms and mites “holobionts themselves interacting with one another”? Microbes inside of the microbiome interact within one another, and can be hosts to other microbes and viruses (see Bdellovibrio and prophage), why isn’t each microbe itself a holobiont? <head briefly explodes>. There has to be a distinction somewhere along the way, and I’m still not getting it even after multiple discussions and reading the PLoS paper. Maybe I’m just prone to thinking everything in Biology is complex and worry too much about the specifics? Even so, there has to be a simple demarcation point going forward that delineates what the hologenome is.

A4b (SB): A4a applies here in addition to the salient difference that while worms live in and outside humans, the worm and human can live without themselves, but neither can live without their microbiomes. This is a key point for why the holobiont and hologenome stop at the host and its associated microbes. We as a community just haven’t appreciated this aspect enough and there’s far more science to delve into because of it.

A4b (KRT): I agree with SB’s broader sentiment here, and will add that the hologenome concept explains (or seeks to explain) the evolution of animals and plants, organisms (i.e. holobionts) whose phenotypes are necessarily products of interactions between their own genomes and those of their symbiotic microbes. The concept does not explain the evolution of microbes.

Q5: What about microbes that are completely neutral to the hologenome? Are these incorporated into the concept (I assume they are)? I’m coining the term “junk microbes” here for these very special cases so that we can have all the fun of the ENCODE folks about arguing what percentage of the hologenome is composed of junk microbes. If you include “junk” microbes into the discussion, then the hologenome includes cases where selection doesn’t act. Therefore, the hologenome idea would include post-reproductive stages of hosts as well, where (hand waving about grandmother’s nurturing effects on evolution aside) the hologenome doesn’t matter for evolution and is just there in the present time for the host. The microbes will of course still evolve with each other. Maybe one prediction is bacterial cheaters arising in older hosts? Just fuel for discussion,  another added layer to think about whether selection is implicit in the concept of the hologenome (I don't think it is).

A5 (SB): Are we really on Q5? Wow :) To your first point, principle VIII and III is all about neutrality and selection in the hologenome. I think we’re on the same page here. And this is an important point of the essay as the original holobiont and hologenome concepts did not distinguish between neutral and selected microbes.

A5 (KRT): Ah, see the “Microbiome Mutiny Hypothesis” (link).

Q6: “Antibiotic or axenic experiments in speciation studies must be routine”. This doesn’t make sense to me, even though I understand exactly where the data is coming from and why you say it. Speciation has never occurred in the absence of microbes in any eukaryote. Taking the microbes away is a completely unrealistic environment and wouldn’t ever happen under natural conditions. I’d be on board with saying microbe swaps are necessary for speciation studies, but to me the axenic experiments don’t tell you anything about speciation in naturo because it’s artificial.

A6 (SB): Would you believe that you disagree with Jerry Coyne too, as he stated the same thing in his Nature review article in 1992! The axenic experiments will importantly tell the relative fraction of reproductive isolation phenotypes that are microbe dependent, something that is assumed by most speciation geneticists to be rare if you were to poll them. So, its a starting point to get us a “symbiotic heritability” if you will for the effects of symbionts vs. host genes on reproductive isolation traits. The microbe swapping is certainly a good next step, but the former is where we lack the most data right now and the easiest to do for wide-ranging systems.

Q7: Phylosymbiosis...Again, I completely get the concept in the broad sense, but this is another place the absolutely needs to be fleshed out. Different branches in the phylogeny should have their own microbiome signals. Where do you draw the line? Microbiomes can easily differ within a species. Microbiomes can easily differ within one individual within one species over time, sometimes in really random ways…….My further guess is that, like most coevolution studies, it’s going to be hard to distinguish between co-evolution and (for lack of a better term) shared environments. Humans/chimps have largely divergent microbiomes, enough to see phylosymbiotic signals. I’m not sure that the same could be said for different species of lemurs on Madagascar because they all fill different niches. Correlations make things messy.

A7 (SB): The question of “where do you draw the line” of microbiome divergence between species for within species is both an experimental question that we’re trying to answer right now and an intellectual question that scales just as well to good old fashion genes. The blurriness in transition from within to between species variation is evidence for evolutionary change over time rather than problematic. Phylosymbiosis is really only best done under “controlled” studies that avoid confounding issues of diet, gender, age, etc. That is why the Nasonia and Hydra studies stand out in my mind. We’ve got more data on other systems as well now. And its important to note here that phylosymbiosis does not equal coevolution. I think we've been pretty clear about this since its inception. I actually came up with the word phylosymbiosis precisely because the concordance between the host phylogeny and microbiome communities does not necessarily mean coevolution.   

A7 (KRT): As the field biologist, or at least once upon a time field biologist, I will suggest that, while I agree with everything SB wrote, broad phylosymbiotic investigations in the wild have merit in that they reveal the extent of microbiota/microbiome variation among hosts in natural systems (descriptive data that are needed), and, if no host phylogenetic signal in symbiotic microbial community composition or structure is evident (while taking into account other host characteristics like diet, sex, etc.), then it strongly suggests that these host-microbial associations are not as pronounced and persistent as the holobiont and hologenome concepts posit.

Q8:  I’m “in like” with the analogy to mitochondria and microbiomes, I’m not in love with it. You can lose some of your microbes and still be functional, you can’t lose your mitochondria and still be a functional human. I think there is a fundamental difference, given phenotypic redundancy in the microbiome. Always willing to be proven otherwise:)

A8 (SB): Right…it's a gradient of dependency. I take the fact that certain microbes can be lost, but some microbes are required as a compelling reason for why the holobiont and hologenome concepts are relevant and novel. We know that the microbes need to be there to some degree and with some specificity, but we also know that any given microbe many not be required. Fascinating!

Thursday, November 19, 2015

This Is What Congressional Support For Science Looks Like

STUNNING! (While it is only for NIH, it is still welcome news.)

Dependent Care Travel Policy for Colleges and Universities

Academia is a profession that can require a substantial amount of travel - from serving on grant review panels to attending conferences and more. While it is a tremendous opportunity to see the world and meet new people, it is also problematic for some. Faculty, staff, and students with families or dependents cannot readily travel, and yet they are often measured in job interviews or tenure and promotion reviews on the extent to which they have spoken at conferences or served on panels. In response to this problem, several universities have earned my respect in leading the way with travel assistance policies. 

I wrote the letter below to bring a similar policy to fruition at Vanderbilt. It is a work in progress as it moves through leadership and adminsitratorion (with some progress), but perhaps this letter will help others in the same pursuit. 


EXAMPLE LETTER                                                                

Dear Colleagues:

Thank you for taking the time to read this proposal on behalf of Vanderbilt University's single and dual-career parents and other dependent caretakers. From research staff to faculty, professionals in academia often struggle to find the work-life balance and financial resources to attend national and international meetings because the costs of airfare and care services for dependents (children, elderly, ill, or disabled family) are prohibitive. As a result, their scholarship is at a disadvantage relative to others who are not juggling dependent care in academia. Indeed, childcare is one increasing and welcome trend at large meetings, yet without the support to travel to conferences, staff and faculty parents must restrict their professional development and networking.

In response to this escalating demand, universities are pioneering Family-Friendly Travel Policies (see five examples below with web links). Each one supports employees with financial assistance for airfare and/or care services for dependents per fiscal year. At a time when single and dual-career parents and caretakers are rising, it seems that a similar policy at Vanderbilt would not only lift the burden of current scholars with families, but also serve as a recruiting tool for those that consider family policies in their job search. Indeed, there appears to be no existing policy after discussing this topic with colleagues in the Schools of Arts and Science, Law, Medicine, Nursing, and Peabody. A reduced opportunity to speak, meet colleagues, and hear about the latest work in the field translates for many to reduced career growth. We can eliminate this bias and at the same time lift the spirit around family-friendly policies at Vanderbilt. 

Ø  Cornell University | Faculty Dependent Care Travel Fund | $1000 per year
Ø  UC Berkeley | Dependent Care Travel Policy | Covers all costs + 70% more than actual costs to offset taxes that will be deducted
Ø  Northwestern University | Dependent Care Professional Travel Grant Program | $750 per year
Ø  University of Michigan | Child Travel Expense Policy | $1000 per year
Ø  Brown University | Dependent Care Travel Fund | $750 per year

Thank you very much for considering this Faculty Life principle.

Wednesday, October 28, 2015

Important Message About Funding Postdocs

I received this university-wide message today about potentially big changes to how we fund postdocs. Its unclear what will happen, but it is an important message for labs with postdocs.

The Fair Labor Standards Act (FLSA) is the federal law governing minimum wages and overtime pay.  It currently requires that most* employees making less than $23,660 record their hours and must be paid for overtime at 1.5 times their usual rate for any hours worked over 40.  The federal government has proposed a revision to the FLSA which would increase this minimum threshold to $50,440.  This proposed change, which will likely take effect before November 2016, will apply to the wages we are federally required to pay postdoctoral scholars.

Most of our postdocs currently receive salaries of $42,000 or more, and are therefore within roughly $8,000 of the proposed new minimum. Most postdocs regularly work more than 40 hours/week, and it would make financial sense to increase these salaries to the new federal minimum rather than pay overtime.  Yet even for postdocs who do not regularly work more than 40 hours, university leaders believe that redefining postdocs as hourly workers would undermine the professional nature of their work.

Changes to the FLSA, if enacted, would have a significant impact upon department budgets, grant budgets, and more. There are still many details to be clarified, and numerous offices across the university are working to determine possible and best courses of action. The final rule will determine the actions we need to take. In the meantime, we want to be prepared for the likely scenario of needing to increase postdoctoral salaries. 

For the present, please do two things:
1.      Alert all necessary members of your department to take these proposed changes into account as needed (for example, budgeting, writing grant proposals, making new offers to postdoctoral scholars, etc.)  Again, changes will likely take effect before November 2016 but the final regulation has not been published yet. No immediate changes to salaries need to be made at this time.
2.      Provide resources for those who have questions:
a.      The documents attached to the email provide some further information.
b.     XXXXX is the primary contact person for specific questions.  She is available for meetings with faculty groups. 
c.       Remind those with questions that this is an open issue which is subject to change and for which the university is still exploring options.

*The law provides certain exceptions, including executives, MDs, teachers, lawyers, and trainees. Postdocs whose primary duty is teaching could be defined as holding a teaching position and receive this exception. The federal definition of “trainee” includes that trainees may not receive wages, so this designation is not an option for most postdocs with the exception of those on NIH NRSA training grants.

Tuesday, October 13, 2015

My phage talk at North Carolina State University

Just got back from North Carolina State University last week. Eric Miller invited me out for a lecture in his Department of Plant and Microbial Biology. Eric has been involved with various bacteriophage projects over the years, including most recently the HHMI phage genome sequencing project and phages that might be useful for killing bee pathogens. I was really impressed with NCSU and how linked in they are with the various industries of Research Triangle. Nearly every person I spoke to had a collaborative, educational, or financial relationship of some kind with them. I don't often see this kind of collaboration between industry and academia, and it is clearly a model for other universities to follow. As a side note, NCSU just launched a major initiative on hiring new faculty in the area of non-human microbiomes.

Anyway, I presented our long-term studies and recent @srbordenstein work on Wolbachia's phage WO, which has some very interesting features that are unique to the viral world. This work is currently in review. I also covered work by @JMetcalfVU and @DNADiva87 on bioprospecting Archaea for new antibiotics that originally stemmed from our investigations of phage WO (link to paper). Here's the talk that I recorded in Keynote, exported to Quicktime, and posted to YouTube. Comments and questions are most welcome. 

Friday, October 9, 2015

The War on Science

There will always be a war on science. Such is the nature of humanity. This video is worth your next five minutes. It speaks for itself....

"Somwhere, something incredible is waiting to be known" - Carl Sagan

Friday, October 2, 2015

Wolbachia Milestone: "Video Game" Status

There are many reasons why Wolbachia bacteria are one of the most amazing microorganisms that you'll ever read about (and why scientists have studied them for almost a century now). For starters, they curiously dwell inside the cells of animal gonads (testes and ovaries) spanning insects to filarial worms. Second, they can change the gender or reproduction of those insects. Third, they can not only prevent mosquitoes from passing on dengue virus to humans, but they may also be a target for curing diseases such as lymphatic filariasis or river blindness. Fourth, they are one of the best cases of how infectious microorganisms can assist the origin of new species. 

Among many other astounding facts, Wolbachia have become so famed apparently in the popular news that they are also now featured in the video game series Metal Gear Solid 5. A friend pointed me to this sound track yesterday with the quote "can't escape it - Wolbachia are now making appearances in video games. Wolbachia in this game are used by the enemy to create a pathogen that kills people speaking only certain languages". 

The full video / sound track is here:

The two clips where Wolbachia are featured are:

and here:

Sunday, July 12, 2015

PeerJ has 5 Nobel Prize Winners on Editorial Board

Our lab has largely embraced the open access paradigm in recent years by choosing, when possible, to publish in open accesss journals. For our highest quality work, we typically submit to PLOS Biology  (Mom Knows Best by @DNAdiva87 and Host Biology in Light of the Microbiome with @KevinRTheis) and eLife (Antibacterial Gene Transfer by @JMetcalfVU). For other papers in the last couple of years, we supported PeerJ (our articles here) - a disruptive, open access publisher and award winning biological and medical sciences journal that publishes reviews and rebuttal letters alongside each article, is fully transparent in the review process, and is importantly affordable. Imagine my surprise today then when I searched the editorial board and found FIVE, count them yourself below - FIVE!!, Nobel Prize winners as well as colleagues Jonathan Eisen and Irene Newton. Anyone else blown away? If so, then support PeerJ.

Friday, June 12, 2015

What are Holobionts and Hologenomes?

Updated 08/09/16:

1. The article referred to below is now published at PLOS Biology (Bordenstein and Theis 2015)

2. Holobionts are defined as the host plus of all its microbial symbionts, including transient and stable members. The term was originally and briefly defined in 1991 by Lynn Margulis in the book Symbiosis as a Source of Evolutionary Innovation (link) as a "compound of recognizable bionts". Bionts = organisms. The bionts can be cooperative or competitive. Please see the aforementioned PLOS article for details.

3. Hologenomes are then defined as the genomes of the holobiont, i.e., the host and microbial genomes, and the pluralistic attributes of a holobiont scale directly to the hologenome (Theis et al, 2016, mSystems). The terms were first used by Richard Jefferson in 2004 (link) and independently by Eugene Rosenberg and Ilana Zilber-Rosenberg in 2007 (link).

June 12, 2015
For the past eight months, Kevin Theis and I have been working hard on an essay on holobionts and their hologenomes. These terms and concepts are well known to a small cadre of biologists, but otherwise are mired in confusion and misinterpretation in part because the literature in this area is diffuse and sparse. Our paper has now gone through a mostly transparent review, and in and outside of that process, we have received some excellent feedback from colleagues in the community. I'm particularly interested in scaling the feedback up and crowdsourcing the process through the comments section below. Some of these principles may not make sense to the reader because we have not yet put forth the entire manuscript (due to be published this year). Nonetheless, I hope that some of what's below resonates with readers to stir a lively discussion. Thanks in advance for your perusal and feedback if you can. This is a draft in progress that can only improve with your insights. 

I. Holobionts and hologenomes are units of biological organization

o   Complex multicellular eukaryotes are not and have never been autonomous organisms, but rather are biological units organized from numerous microbial symbionts and their genomes
o   Biomolecular associations between host and microbiota are more conceptually similar to an intergenomic, genotype x genotype interaction, than a genotype x environment interaction

II. Holobionts and hologenomes are not organ systems, superorganisms, or metagenomes

o   As holobionts are complex assemblages of organisms consisting of diverse microbial genomes, biology should draw a clear distinction between holobionts/hologenomes and other terms that were not intended to describe host-symbiont associations
o   Organ systems and superorganisms are biological entities comprised of one organism's genome; Metagenome means "after" or "beyond" the genome, does not intrinsically imply organismality for environmental samples, and obviates the obviates the fundamentals of symbiosis in the holobiont

III. The hologenome is a comprehensive gene system

o   The hologenome consists of the nuclear genome, organelles, and microbiome
o   Beneficial, deleterious, and neutral mutations in any of these genomic subunits underlie hologenomic variation

IV. Hologenomic evolution is most easily understood by equating a gene in the nuclear genome to a microbe in the microbiome

o   Evolution for both genes and symbionts is fundamentally a change in population frequency over successive generations, i.e., the fraction of holobionts carrying that particular nuclear allele or microbe.
o   Covariance of hosts and microbes in a holobiont population (i.e., community genetics) follows a theoretical continuum directly to coinheritance of gene combinations within a genome (i.e., population genetics)
o   A grand unified theory of evolutionary and ecological genetics deserves priority attention

V. Hologenomic variation integrates all mechanisms of mutation

o   Every hologenome is a multiple mutant meaning that there is variation across many individual genomes spanning the nucleus, organelles, and members of microbiome
o   Base pair mutation, horizontal gene transfer, recombination, gene loss and duplication, and microbial loss and amplification are all sources of variation

VI. The hologenome concept reboots elements of Lamarckian evolution

o   Although Lamarck never imagined microbes in his theory, applying the tenets to holobionts rebirths some major aspects of Lamarckism
o   The nuclear genome is inherited mainly within a Mendelian framework, but the microbiome is originally acquired from the environment and may become inherited
o   Host-microbe associations can forge disequilibria via parental transfer or stable environmental transmission

VII. The hologenome concept fits squarely into genetics and accommodates multi-level selection theory

o   Multi-level selection theory asserts that selection operates across multiple levels of genetic variation with phenotypic effects, from genes to hologenomes and beyond
o   Holobionts are exclusive to hosts and their associated microbiota. Different holobionts, such as a pollinator and flower, interact with each other under standard ecological principles

VIII. The hologenome is shaped by selection and neutrality

o   Natural selection can work to remove deleterious nuclear mutations or microbes, while spreading advantageous nuclear mutations or microbes. In the absence of selection, the neutral spread of hologenomic variation through populations is an inherently stochastic process
o   Mixed ecological models of stochastic and deterministic community assembly likely reflect natural systems, and partitioning the microbiota into stochastic versus deterministic subunits will be an important future goal of the field

IX. Hologenomic speciation blends genetics and symbiosis

o   The Biological Species Concept was never intended to be exclusive of symbiosis, though history largely divorced the two and created unnecessary controversy
o   Antibiotic or axenic experiments in speciation studies must be a routine, if not obligatory, set of experiments in genetic analyses of speciation for an all-inclusive understanding of the origin of species.

X. Holobionts and their hologenomes do not change the rules of evolutionary biology

o   There is no fundamental rewriting of Darwin's and Wallace's theory of evolutionary biology, though the concepts redefine that which constitutes an individual animal or plant

o   Simply put, if the microbiome is a major, if not dominant, component of the DNA of a holobiont, then microbiome variation can quite naturally lead to new adaptations and speciation just like variation in nuclear genes