Application of new DNA sequencing methods may determine the cause of a disease that is enigmatic but plaguing sensitive coral reefs
We are trying to find the biological culprit behind the 2012 Barrell Sponge (Xestospongia muta) Die Offs
A marine epidemic called “small orange band” (SOB) disease of the common yet iconic "barrel" reef sponge, Xestospongia muta, re-emerged on South Florida sponges in April 2012. Gross SOB symptoms appeared as bleaching, decaying and crumbling of the affected sponge tissues, followed by rapid death of the whole or most of the sponge individual. The photo below shows how the sponge was almost completely "dissolved" by this phenomenon. During this outbreak, anecdotal reports and video surveys (like those shown above) carried out by REEF-RESCUE.org found that 15- 20% of local large barrel sponges died or were adversely affected. The SOB condition has been witnessed periodically, and has been histologically described in scientific studies. However, the question remains "What is the cause of this blight and destruction?" YET TO DATE, NO CLEAR CAUSE OR PATHOGEN HAS BEEN IDENTIFIED. This condition could appear again, but with further research on the possible cause we could be better prepared to handle it.
In May 2012, our laboratory was able to collect multiple samples of diseased and healthy sponges for in depth genetic and microscopic analyses. We hypothesize that current molecular genetics biotechnologies may be able provide basic answers to what factors are possibly causing SOB.
Recent studies and breakthroughs in DNA and RNA sequencing technologies are revealing the inner mechanisms of how our cells and bodies work. Sponges are the most primitive animal form (http://en.wikipedia.org/wiki/Sponge), but they may still educate us on fundamental biological concepts, such as the role of microbial symbioses (foreign cells that living with larger organisms but causing no harm). This project will apply lessons learned from recent symbiosis studies such as the current Human Microbiome Project (HMP)- http://www.hmpdacc.org/impacts_health/impact_health.php. The HMP applies "high throughput" DNA sequencing methods to characterize the thousands of microbes that live in and within our bodies, often to our benefit. For example, some bacterial species benignly live in our intestinal tracts and help digest plant materials, or provide essential vitamins that our bodies cannot produce.
These molecular biotechnology approaches can be applied to REEF studies and may indirectly help PROTECT REEFS. Coral reef ecosystems are in peril due to many threats, such as pollution, global warming and disease - http://www.nytimes.com/2012/07/14/opinion/a-world-without-coral-reefs.html?_r=1&emc=eta1. However, previous appeals to various agencies have not yet resulted in funding for this project.
Until adequate funds are obtained, the Xestospongia muta samples we have collected will remain frozen, archived and unanalyzed. Thus, this is a direct appeal for funds that will be carried out by our Marine Microbiology and Genetics Laboratory in the brand new Center of Excellence for Coral Reef Ecosystems Sciences of Nova Southeastern University (NSU), in Dania Beach Florida (www.nova.edu/ocean). This is a state of the art facility with modern molecular labs to conduct the work. NSU is a non-profit university, and 100% of donations for this project will be used towards the science or the rewards related to the science.
The Number 1 goal of this project is to understand and determine the cause of SOB disease on Xestospongia muta. Funds received from this project will go to the following experimental activities:
1. To apply the latest high tech “next generation” DNA sequencing methods to characterize the microbial diversity (eubacteria, archaea, eukaryotes, and viruses) which may be associated with healthy and disease Xestospongia muta sponges. We have already characterized sponge microbial partners in a recent paper by White et al 2012 (see reference 2 below for downloadable PDF).
2. To evaluate the effects of the disease on host physiology, Illumina HiSeq of total RNA (cDNA) will be performed on the samples representing the three physical states of the sponge (healthy, diseased, healthy sections from diseased animals). Current costs for “Next Generation” Illumina HiSeq 100 bp DNA sequencing that we will be applying, can be viewed at http://www.ibbr.umd.edu/facilities/sequencing/illumina/pricing. Almost half of the total costs of the project will be to prepare RNA libraries for each sample (N=25), which requires great care due to the fragility and low amounts of "messenger RNA" that will be sequenced.
The photo above shows the current Illumina DNA sequencer that will be used to read the extracted sponge samples.
Other costs to be covered will be a) fixing sponge samples for electron microscopy and b) the application of analytical software involved in analyzing the large volumes of data that will be generated by Illumina DNA sequencers. The data will culminate in at least one peer-reviewed publication(s), thousands of new gene sequences uploaded to the public GenBank data repository: http://www.ncbi.nlm.nih.gov/guide/.
3. We also aim to work closely with other local and state agency groups concerned about the welfare of reefs, such as http://research.myfwc.com/ to keep them informed of advances.
Our research team will greatly appreciate any support you can provide, and will highlight and fully acknowledge your donations with the rewards we will provide upon reaching our funding goal.
For Further Reading:
1. Angermeier H, Kamke J, Abdelmohsen UR, Krohne G, Pawlik JR, Lindquist NL, Hentschel U. (2011) The pathology of sponge orange band disease affecting the Caribbean barrel sponge Xestospongia muta. FEMS Microbiol Ecol. 75: 218-30.
2. White, J, Patel, J., Ottesen, A., Arce, C., Blackwelder, P., Lopez, J.V 2012.Pyrosequencing of microbes within Axinella corrugata sponges: Diversity and seasonal variability. PLoS ONE 7(6): e38204. doi:10.1371/journal.pone.0038204; http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0038204
3. Webster, N. S., A. P. Negri, R. I. Webb and Hill R. T. (2002). A spongin-boring a-proteobacterium is the etiological agent of disease in the Great Barrier Reef sponge, Rhopaloeides odorabile. Mar. Ecol. Prog. Ser. 323:305-309.
4. Vega Thurber, R.L. and Correa. Viruses of reef-building scleractinian corals. (2011). JEMBE 408:102–113
5. Human Microbiome Project Consortium with 248 Collaborators. Structure, function and diversity of the healthy human microbiome. Nature. 2012 Jun 13;486(7402):207-14. doi: 10.1038/nature11234.
Have a question? If the info above doesn't help, you can ask the project creator directly.
pledged of $17,600 goal
seconds to go
Funding Unsuccessful This project reached the deadline without achieving its funding goal on October 12, 2012.
Sep 2, 2012 - Oct 12, 2012 (40 days)
Pledge $20 or more
I will send a personal thank-you email to anyone who donates $20 or more to this project. The Thank you email will be a genuine expression of my gratitude for supporting this project.Estimated delivery: Oct 2012
Pledge $70 or more
I will contact you by phone or SKYPE to personally thank you, and provide a 30 minute overview of the project, followed by a personalized letter of thanks and photograph of the reefs to mark our discussion and your supportEstimated delivery: Oct 2012
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At a mutually agreeable time, I will lunch with you and give a 2 hour detailed tour of my laboratory at the Center of Excellence in Coral Reef Ecosystem Sciences at Nova Southeastern University, Dania Beach, FL – www.nova.edu/oceanEstimated delivery: Oct 2012
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I will present a one-hour original seminar on the research project. I will then deliver the seminar at a mutually convenient time. The seminar can be followed by a half-hour discussion. All travel and accommodation expenses are the responsibility of the donor.Estimated delivery: Nov 2012
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I will place a formal acknowledgement in the peer-review publication resulting from the project thanking either the individual or the organizational donation. The acknowledgement will read “We thank ________! for supporting this research through a generous donation on Kickstarter”.Estimated delivery: Jan 2014
Pledge $1,600 or more
Dive with the scientist. I will accompany anyone with a valid open water diving permit, and current DAN insurance, to a local (Ft Lauderdale) Florida coral reef to view the amazing habitats and model organisms. This will include boat costs, but you will have to bring your own SCUBA equipment. Similar to the previous reward you will also be acknowledged in any peer reviewed publication that utilized your donations.Estimated delivery: Oct 2012