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Post by safrey on May 29, 2007 19:41:11 GMT -5
Lastnight I collected a living specimen from my hair for the first time. I observed this specimen at 200x magnification and, other than the enviroment in which it is living, it exhibits charcteristics exactly like those of a bryozoan, including the "flicking" of tentacles, a classic bryozoan trait, it also has brilliant green eyes, which I have yet to find reference to in regards to bryozoans. I have video of this specimen if anyone would like to see it but have not uploaded this video anywhere as of yet. Any suggestions as to where I may post it?
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Post by Admin on May 29, 2007 20:11:21 GMT -5
Steve........Thanks for trusting us with your personal involvement here. We were not aware that you and your family are victims of this horror...........
What Whiterose is saying is that we have a researcher, Dr. Hildegarde Staninger, who hs spent the better part of a year researching the Morgellons fibers and we have lab results which were presented with a 3 hour radio audio.
We also have silicone running wild in Jan Smith's knee tissue and fibers of High Density Polyethlyene.
We have images of fibers which, when burned, show a glowing Golden head attached to a blue fiber which shows circular arrays within the fibers......Can only be manmade.
I will bring the links all over here - and hope that you spend the time listening to Audio Specials No. 5, 6 and 7 on rense.com they are all there - - three - three=hour shows...and the documentation for each one needs to be followed on the internet pages while listening........as in a show and tell ..........
- since it is OBVIOUS to me you are not only a brilliant researcher - but needful, as are the people on this board - for more answers -If you are willing to do that, I/we may be able to all get on the same page. There is certainly more power in numbers of minds working together.
Welcome, here, Steve.....Blessings, Shoshanna
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Post by Admin on May 29, 2007 22:28:13 GMT -5
Dr. Hildegarde Staninger - Home drhildy.proboards46.com/index.cgi RENSE MORGELLONS RADIO SPECIALS -------------------------------------------------------------------------------- MORGELLONS SPECIAL No. 1 Cliff Mickelson & Jan Smith Morgellons Victims June 22, 2006 (MP3 Audio File) www.cherokeechas.com/mp3/CM-rns01.mp3 MORGELLONS SPECIAL No. 2 Cliff Mickelson, Jan Smith, Cindy Casey, RN and Charles Holman Morgellons Victims June 27, 2006 (MP3 Audio File) www.cherokeechas.com/mp3/CM-rns02.mp3 MORGELLONS SPECIAL No. 3 Cliff Mickelson, Jan Smith, Cindy Casey, RN and Charles Holman, Greg Smith, MD & Judy Smith, RN Morgellons Victims July 17, 2006 www.cherokeechas.com/mp3/CM-rns03.mp3(MP3 Audio File) MORGELLONS SPECIAL No. 4 Cliff Mickelson, Jan Smith, Gill from Melbourne, Australia Morgellons Victims August 18, 2006 (MP3 Audio File) www.cherokeechas.com/mp3/CM-rns04.mp3 Courtesy of New Morgellons Order www.cherokeechas.com/ MORGELLONS SPECIAL No. 5 Hildegarde Staninger, PhD, RIET-1 Rahim Karjoo, M.D., F.A.S.C.P./F.C.A.P. Jan Smith, Morgellons Victim, Stunning Data, Solid Evidence October 24, 2006 (MP3 Audio File) rense.gsradio.net:8080/rense/special/rensemorgellons5.mp3 Documentation-Morgellons Disease-Remarkable New Lab Findings No. 5 www.dldewey.com/morgel.htm MORGELLONS SPECIAL No. 6 Hildegarde Staninger, PhD, RIET-1 Rahim Karjoo, M.D., F.A.S.C.P./F.C.A.P. Jan Smith, Morgellons Victim Shocking Breakthroughs! November 9, 2006 (MP3 Audio File) rense.gsradio.net:8080/rense/special/rensemorgellons6.mp3 Documentation: Morgellons Specimen Photos in Sequence - Special No. 6 www.rense.com/general74/morg6-1.htm MORGELLONS SPECIAL No. 7 Hildegarde Staninger, PhD, RIET-1 Dr. R. Michael, Castle Jan Smith, Morgellons Victim Morgellons Identified - World Exclusive! Shocking Research - Nanotech Machines In The Human Body March 29, 2007 (MP3 Audio File) rense.gsradio.net:8080/rense/special/rensemorgellons7.mp3 Morgellons #7 Documentation Project FMM - Lab Report Analyses - Dr. Staninger (49 pages) www.rense.com/morgphase/phase2_1.htm MORGELLONS SPECIAL No. 8 Psych, Emotional, Personality And Relationship Impacts - Cliff Mickelson, Jan Smith & Guests May 10, 2007 (MP3 Audio File) rense.gsradio.net:8080/rense/special/rensemorgellons8.mp3 -------------------------------------------------------------------------------- See what's free at AOL.com.
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Post by Admin on May 29, 2007 22:31:14 GMT -5
Please notice to COPY AND PASTE the entire URLs into your browser for the rense.com audio files in Special Nos. 5, 6, 7 & 8.
Our system here is not supporting making a hotlink.
Thanks.....Sho
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Post by safrey on May 29, 2007 23:14:36 GMT -5
I have already read most of what you've posted regarding Dr Hildegarde Staninger work and I think that she is very talented and sincere in her efforts. The results of the chemical analysis she has performed play an important role in my hypothosis as well. I would be interested to know if she has ever looked at the chemical analysis of sponge and bryozoan spicules. I am not a chemist by any stretch of the word but from what I find the two(morgellons fibers and sponge spicules) appear very similar in their composition. Chemical Composition of a Spicule
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Post by skytroll on May 30, 2007 1:27:28 GMT -5
Safrey,
Am so sorry to hear your family has this.
We are trying to pitch together, here, and many of us have differing ideas, but, we do want so much to get to the real facts, here.
Dr. Staninger has some facts, some observations, which are in the above links.
You say you saw them, do you think these spicules may be being used for other purposes?
If there is an axial filament composed of protein, could those be substituted into the human body and be used for oh, maybe artificial muscles? or nerves or even capillaries?
How do they get in the human body? This is what I wonder?
Am tired, but, will read more on your posts, later, and welcome to the board. We all try to aim in a somewhat coherent fashion. Would you think that nano machines could mimic bryozoans? Also if used for fiber optics, could these also be used in humans?
My big question is , though, is how would these get into humans and how would they survive there?
Can the silica be used as nano bricks? Seems there is an exploitation of many proteins common in organisms to be used for other purposes, even for human medicine.
Take care,
Hopefully after some sleep, I can contribute more.
Hang on folks, there is something about the silica and carbon there. mmmmmm
Well, lets keep at it.
Thanks Safrey.
Skytroll
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Post by Admin on May 30, 2007 2:19:54 GMT -5
"For Demospongiae it has been well established that the siliceous spicules have a central core and an axial filament composed of protein. Important is the observation that these filaments are impregnated with silica. Specialized cells, the sclerocytes, which are characterized by a high metabolic activity, secrete spicules. Secretion of spicules appears to proceed either intracellularly or extracellularly, very likely depending on the size of the spicules"
The fibers from our M's victims have no core - they are like straws...and some like a straw covered with a polymer coating.
Sure would like to see more images.....
(about your video - can you figure out how to post it on YouTube? There are a host of M's victims' specimen videos there.)
I only saw one image - and it looked like a SPONGE (yup!) with fibers growing out of the end up toward the body of the SPONGE.... Have not seen any M's victims with specimens like that.
Tired, too.......and will come back to work here tomorrow.
Most of the images we see are "..........-like" in that they resemble thngs we 'know' but are off somewhat - as in mutated and/or spliced DNA creatures......and as a result, synthetic.
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Post by safrey on May 30, 2007 20:30:45 GMT -5
Thanks for the welcome. I just want to say that I am here for the same reason we all are, to put an end to this nightmare. Let me tell you where I stand. I can contribute significantly to the investigation of the morgellons parasite in many ways. I have dealt with the symptoms for over 13 years, performed numerous experiments, made countless microscopic observations(up to 200x), and have researched extensively in the field of invertebrates, and a substantial amount in the fields of bacterias, minerals and elements, and a small amount in nano-technology as they would apply to this investigation. I believe I may be one of the only, if not the only one to have witnessed and documented the ability of this organism to transcend a latex glove (which I have an explanation for), I have grown the same culture that the silentsuperbug video illustrates and observed and documented the formation of an extremely complex crystal that I believe is comprised of calcite(calcium carbonate), this crystal is so unusual that it alone compelled me to research into nano-technology and to this day I am still not convinced that nature could create something like this. On the other hand I have much more compelling evidence to conclude that in some way the traits and characteristics of a bryozoan are at work here. One of the most compelling pieces of evidence is in reference to the photo # 5 at rense: Morgellon's Photos 5www.rense.com/general74/morg6-5.htmthis is the classic description of an "avicularia" which is a life form of the bryozoan. Full text here: www.flmnh.ufl.edu/malacology/IZ2005/LabNotes/Lab10/lab10.htmThere are many more correlations that can be made between morgellons and the bryozoan including Cliff's and other detailed descriptions of observations and the decription of a zooid and bryozoan colony. I'm sorry for re-referencing this but it's very important. Partial description of a bryozoan Full text of this article contained here: earthlife.net/inverts/bryozoa.htmlFull text of this article contained here: www.reefkeeping.com/issues/2003-10/rs/index.phpExcerts from Cliff's and Chester's observations These are just some of the correlations, and there is much more evidence to support this claim, but that does not mean that I have ruled out nano-technology completely, the crystal still needs an answer. The little research I did into nano-technology taught me three things relative to this investigation. One, Calcite, which I'm certain is a factor here for numerous reasons which I can go into at another time, is not only applicable to the bryozoan but to nano-technology as well. Two, there has been and currently is research being performed by those involved in nano-technology involving the sponge and it's ability to produce silica fibers like those in fiber-optic industry. Three, I believe the object in nano-tech is generally to replicate something that is produced by nature, correct me if I'm wrong here, the bryozoan could be choosen for several reasons. Now I am just speculating here but if I were trying to replicate something for say sewage treatment or oil spill cleanup I might consider the bryozoan because it has the ability to ingest chemicals, they're part of it's diet. So I think that it's plausible that something like this has taken place and has had unexpected consequences. This being said, I am not implying that I believe this to be true only suggesting that if evidence pointed in that direction it could fit into my hypothesis. So in my opinion, for whatever it's worth, is that any research done to connect this with nano-technology should include the bryozoan because there is simply so much evidence indicating it's involvement. One last thing, the bryozoans tentacles are hollow, and very complex, they deliver sperm through them to the egg. It is my contention that the one of the adaptations that the bryozoan has made, or someone has made, for it's different enviroment is to it's tentacles, particularly their length and function. I think that some of the fiber material we find with morgellons may be bryozoan tentacles, this is an area that I haven't pinned down completely but I'm leaning towards the blue fibers as the ones. Do you know if all of the fibers analyzed are hollow? This is all for now been a long post so I will try to answer your other questions later
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Post by skytroll on May 30, 2007 22:30:32 GMT -5
Wow......Safrey........the calcite........I am interested in.......and that could be involved, but, could it be the bryozoan dying or replicating itself?
could a synthetic gene have been made that would mimic the bryozoan?
The nano then would be synthetic, but the calcite could be part of the grown or death of the bryozoan.
see what I can find.......
Thank you for clarifying that again,
Do you think this tech is heading for biocomputer or human as computer?
Or do you think this was a microbial mistake by the Bioremediation folks?
Skytroll
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Post by skytroll on May 30, 2007 22:39:21 GMT -5
safrey, did you see this? Note the critter going back into the sheath. tinyurl.com/35mhvkskytroll
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Post by skytroll on May 30, 2007 22:41:41 GMT -5
However how can something that looks like dirt, start moving and do this?
Has to assembly or is incognitio. hiding inside of ......say a nanotube?
SKytroll
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Post by safrey on May 30, 2007 23:41:01 GMT -5
from what I've found calcite seems to be something they excrete continuously, other invertebrates do as well to make their exoskeletons, they produce it. there are many aspects of morgellons that suggest calcite or calcium carbonate, like for one the orange glow under ultraviolet that some are reporting seeing in nasal passages, I personally have orange between my toes(worst of my morgellons affected areas) under ultraviolet, calcite glows orange under ultraviolet.
this I don't know, I know they are working with the sponge in that respect but the bryozoan is a bit more complex.
after learning just how bizarre the bryozoan is by nature I tend to lean more towards an all nature cause, with two exceptions, one I already mentioned that being the crystal, the other is just how bizarre the eyes are on this specimen that I collected and videoed the other night, you have to see them, brilliantly green like they are lit up, very strange, the video is 42 meg long and I have dial-up so need to upload from some other location
I am still waiting for this video to load, takes forever with dial-up but "going back into the sheath" sounds like something a bryozaon would do, will comment later.
There is one trait that can explain many of the very odd occurrences in morgellons. It is a trait that is, based on my research, thought to only exist in sponges(I cannot find reference to other animals having this trait), that trait is the ability to reconstruct itself. They say if you put a sponge in a blender, mix it up and pour it into a bowl that it can put itself back together! This can also explain one of Cliff's findings,
To me this sounds exactly what a creature with the ability to reconstruct itself would do, to transcend a latex glove or to change shape, although science knows of I think six forms of the bryozaoan but could there be a limit? I mean, a creature that can break down into pieces and putt itself back together, thats a pretty bizarre creature.
Since the sponge and bryozoan share other traits that are unique between the two of them I am conjecturing that the bryozoan also shares this trait and science just has not documented it yet, there is a great deal unknown about the bryozoans, especially a small group called burrowing bryozoans.
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Post by skytroll on May 31, 2007 1:15:20 GMT -5
Now, if these are burrow, don't they burrow into coral, or sponge? Why would they want to burrow into humans, when they usually burrow into water bearing plants, like algae, or even cyanobacteria, but can they live in spores? mmmmmmm
Interesting little fellas.
Skytroll
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Post by sarahconnor on May 31, 2007 3:17:30 GMT -5
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Post by whiterose on May 31, 2007 10:50:06 GMT -5
Something that should be kept in mind is that there are different variants of morgellons, I have heard as many as 11. Just something to consider.
wr
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Post by safrey on May 31, 2007 19:10:47 GMT -5
Burrowing bryozoans are know to burrow into crustaceans, living within their shell, multiplying until they eventually kill the organism within. This is a quote from an article co-authored by two of the most knowledgeable people in the world regarding bryozoans, JOHN D. SOULE and DOROTHY F. SOULE. icb.oxfordjournals.org/cgi/content/abstract/9/3/791If you think about it, what would be the difference between living within a shell or living with a human body?
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Post by whiterose on May 31, 2007 20:47:01 GMT -5
I'm so sorry, I spelled it wrong, it is Raman not Ramon, my apologies and here is the link of all the research that was paid for that tells of the Raman scope: www.rense.com/morgphase/phase2_1.htm
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Post by Admin on May 31, 2007 23:49:54 GMT -5
Steve - I have spent quite a number of hours researching for images of Bryozoan - and find absolutely NOTHING that is a match for the 5000+/- specimen images I have from victims.
The 'hook' looks like a claw from a big cat....found that last night, copied it out - downloaded a program which told me I could upload images - and then download them on here - WOW! Pictures! But it did not work.......
Lost the url after uploading the image to the program...didn't think I neede it......duh!
Your research here is extensive - and even the parasitologist experts with whom I have corresponded over the last year - sharing images and asking for their 'take' on what we're looking at - cannot say 100% about ANYTHING until and unless they do DNA testing........ Do you have specimens from your own body that match any Bryozoan images? Can you provide us with links? and just say "I HAVE THIS SAME THING in a bottle!" or some-such, oor post some images on youtube?
-- AND if there is anyone here who knows how to upload images to this Board - PLEASE HELP!
That would help a bunch.......the description of an elephant in words? .... well - you know.
ALSO, you must have an idea as to HOW and at WHAT STAGE these Bryozoan critters may have entered the bodies of M's victims - and some idea as to how thousands are now afflicted from Coast to Coast and around the World.
You say the sponge can self-assemble....or re-generate, if you will. What about the other 'forms' in the Bryo's life cycle?
More later, Steve........ Blessings, Shoshanna
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Post by safrey on Jun 1, 2007 2:12:13 GMT -5
Hi Shoshanna, It was not an overnight process for me to come to this conclussion, it's not cut and dry unfortunantly, For one there is so very much data out there yet still so much unknown. Images of sponges and bryozoans are usually either of colonies rather than an individual zooid and they vary so much. Let me give you a little more insight to my current battle. Earlier in my investigation I had come to the conclusion that the Sponge, or Phylum Porifera, was involved. Then after further study I became convinced it is the bryozoan, now I'm battling a problem. There's a large amount of evidence to suggest that the bryozoan is involved, but yet there is also substaintial evidence that the organism involved has the ability to rebuild itself if seperated, I am having difficulty in tying this ability to the bryozoan, only the sponge can do this as far as I can tell. So tonight I read where it has been found that bryozoans and sponges can be symbiont with each other(an organism that is associated with another in a mutually beneficial relationship). Now a whole new field to research, it never ends Thsi will give you idea what I'm up against as far as diversity Polyzoa is another name for bryozoa Polyzoa from the Gilbert Islands. www.nhm.ac.uk/hosted_sites/iba/bryozoa_home_page/library/maplestone/map1909.htmlJust one type of avicularia www.flmnh.ufl.edu/malacology/IZ2005/LabNotes/Lab10/Image3.jpgI know it's a lot to take in, but I'll throw it at you best I can. I have some specimens on slides yes, they are either partial zooids that I collected from surface of latex glove after they transcended or in a couple I think the whole zooid came thru and reconstructed or whatever it did to get thru.
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Post by safrey on Jun 1, 2007 3:44:16 GMT -5
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Post by whiterose on Jun 1, 2007 5:16:00 GMT -5
So considering everything I have read that you have posted, you are saying this could be nanotechnology that is mimicking Bryozoan.
Now that I can buy, we all know nanotechnology can mimick just about anything. We know Dr. Hildegard Staninger and Dr. Rahim Karjoo who sent samples that were collected from different folks off to five different labs and followed complete procedure protocol were told what we at this board were dealing with is nanotechnology. This information came from 5 different labratories. Are you suggesting that they don't know what they are doing and that their methods were flawed. Or are you saying that with your equipment you were unable to pinpoint that it was nanotechnology?
Perhaps what you have is different than what the rest of us at this board are afflicted with, could that be a possibility? I hope you are able to pin point what is wrong with you and your family.
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Post by safrey on Jun 1, 2007 17:27:30 GMT -5
First let me say that in my opinion all of us who have the fiberlike deposits within the skin are infected with the same organism, maybe different variations but the same, whether that be a manmade one or natural one.
I'm not at all suggesting that the professionals working on this problem don't know what they are doing. I'm not suggeting they are wrong. My objective here is to make everyone aware of the specific correlations between morgellons symptoms and the bryozoan and sponge to encourage further research involving one of these phylums.
Like I mentioned earlier, it is my understanding that nano-technology objectives usually arise from the desire to replicate certain traits know to occur in nature in specific organisms, so in seeking links to nano-tech it might be advantageous to do research into areas where nano-tech links with the sponge or bryozoan.
There is much that still remains unanswered or unaccounted for. All I know for sure is that in some capacity the traits and characteristics of the bryozoan and or possibly the sponge are part of this equation, there is just to much evidence not to conclude that. Whether it be a manmade replication of these organisms or just the incredible act of evolution is yet to be known.
In regards to analysis of the fibers done by Dr Hildegarde Staninger, my only comment is, I wouldn't rush to conclussions, nature is very complex, we learn this lesson all to often.
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Post by whiterose on Jun 1, 2007 19:12:09 GMT -5
You have a right to your opinion sir, but you have no right to categorize anyone else here in with your opinion, that is a bit arrogant on your part.
Your demeaning of Dr. Hildegarde Staninger's work says a great deal of what you are about; furthermore, I'm sure all the lab folks would be interested in your demeaning of their work. This work did not come from a single source but from a group of many professionals, good day sir.
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Post by safrey on Jun 1, 2007 20:09:35 GMT -5
To be honest with you I have no idea how you are reading any of that in my posts. These folks you are referring to may fully believe they have the answer, maybe they do have the answer, maybe they don't. Just because someone claims something is that cause to take it for gospel?
I do know one thing for certain, working with attitudes like I've seen here is not what I'm looking for so good luck in your quest.
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Post by skytroll on Jun 6, 2007 18:39:33 GMT -5
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Post by safrey on Jun 6, 2007 21:13:13 GMT -5
Most interesting articles, thank you for the links. I'm happy to know that atleast your curious enough to research, keep up the good work.
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Post by skytroll on Jun 7, 2007 1:09:22 GMT -5
Well, it seems to me this should be of interest especially the parchment worms, wonder why so many of them in one place.
And they look very strange.
But the bryozoan is part of the lineage to the human genome. That kinda sparked my interest and the connection between synapses and the sponge. I find that qutie interesting.
I look under all the rocks, per se, and look at all directions and also observe these critters on my skin, and observe the environment also.
Thank you for taking a peak.
Skytroll
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Post by safrey on Jun 7, 2007 3:24:02 GMT -5
The more you learn about the bryozoans the more interesting they become, adaptable little animals, seems as though they can make just about whatever adaptation they need to. There is also a direct connection to certain bacteria associated with morgellons: "Candidatus Endobugula glebosa," a Specific Bacterial Symbiont of the Marine Bryozoan Bugula simplex aem.asm.org/cgi/content/abstract/70/8/4921www.springerlink.com/content/x62621703817x2h9/
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Post by skytroll on Jun 7, 2007 7:29:20 GMT -5
why can't we post on this thread? Was knocked off. skytroll
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Post by skytroll on Jun 7, 2007 7:53:09 GMT -5
As I was saying...... Vertical transmission through the sponge of microbes: "In summary, it could be shown that in I. felix, vertical transmission of microorganisms through the larvae is an important mechanism for the establishment of the sponge-microbe association. " aem.asm.org/cgi/content/abstract/73/7/2067-------------------------------------------------------------------------------- mmbr.asm.org/cgi/content/abstract/71/2/295"We review the status of this important field, outlining the various approaches (e.g., cultivation, cell separation, and metagenomics) which have been employed to access the chemical wealth of sponge-microbe associations..... tinyurl.com/39a48gMetagenomics: "Metagenomics (also referred to as environmental and community genomics) is the genomic analysis of microorganisms by direct extraction and cloning of DNA from an assemblage of microorganisms. The development of metagenomics stemmed from the ineluctable evidence that as-yet-uncultured microorganisms represent the vast majority of organisms in most environments on earth. This evidence was derived from analyses of 16S rRNA gene sequences amplified directly from the environment, an approach that avoided the bias imposed by culturing and led to the discovery of vast new lineages of microbial life. Although the portrait of the microbial world was revolutionized by analysis of 16S rRNA genes, such studies yielded only a phylogenetic description of community membership, providing little insight into the genetics, physiology, and biochemistry of the members. Metagenomics provides a second tier of technical innovation that facilitates study of the physiology and ecology of environmental microorganisms. Novel genes and gene products discovered through metagenomics include the first bacteriorhodopsin of bacterial origin; novel small molecules with antimicrobial activity; and new members of families of known proteins, such as an Na+(Li+)/H+ antiporter, RecA, DNA polymerase, and antibiotic resistance determinants. Reassembly of multiple genomes has provided insight into energy and nutrient cycling within the community, genome structure, gene function, population genetics and microheterogeneity, and lateral gene transfer among members of an uncultured community. The application of metagenomic sequence information will facilitate the design of better culturing strategies to link genomic analysis with pure culture studies. " ..."The outcomes of the recognition of uncultured microorganisms are worthy of examination. One of these outcomes, metagenomics, is further shaping microbiology. Metagenomics has already opened new avenues of research by enabling unprecedented analyses of genome heterogeneity and evolution in environmental contexts and providing access to far more microbial diversity than has been viewed in the petri dish. This review will explore the origins of metagenomics and examine its recent application to microbial ecology and biotechnology. " d"The word metagenomics was coined (69) to capture the notion of analysis of a collection of similar but not identical items, as in a meta-analysis, which is an analysis of analyses (64). (Community genomics, environmental genomics, and population genomics are synonyms for the same approach.) The idea of cloning DNA directly from environmental samples was first proposed by Pace (108), and in 1991, the first such cloning in a phage vector was reported (126). The next advance was the construction of a metagenomic library with DNA derived from a mixture of organisms enriched on dried grasses in the laboratory (71). Clones expressing cellulolytic activity were found in these libraries, which were referred to as zoolibraries, a term that has not been used widely in the field (71). The work of DeLong's group defined the field when they reported libraries constructed from prokaryotes in seawater (136). They identified a 40-kb clone that contained a 16S rRNA gene indicating that the clone was derived from an archaeon that had never been cultured. Construction of libraries with DNA extracted from soil lagged due to difficulties associated with maintaining the integrity of DNA during its extraction and purification from a soil matrix (14, 69, 80, 118) but eventually produced analyses analogous to those from seawater (39, 72, 118). APPROACHES TO METAGENOMIC ANALYSIS Top Previous Next References Metagenomic analysis involves isolating DNA from an environmental sample, cloning the DNA into a suitable vector, transforming the clones into a host bacterium, and screening the resulting transformants (Fig. 3). The clones can be screened for phylogenetic markers or "anchors," such as 16S rRNA and recA, or for other conserved genes by hybridization or multiplex PCR (136) or for expression of specific traits, such as enzyme activity or antibiotic production (39, 44, 61, 78, 87, 88, 91, 93, 118, 125), or they can be sequenced randomly (141, 142). Each approach has strengths and limitations; together these approaches have enriched our understanding of the uncultured world, providing insight into groups of prokaryotes that are otherwise entirely unknown. " "Symbiosis Many bacterial symbionts that have highly specialized and ancient relationships with their hosts do not grow readily in culture. Many of them live in specialized structures, often in pure or highly enriched culture, in host tissues, making them ideal candidates for metagenomic analysis because the bacteria can be separated readily from host tissue and other microorganisms. This type of analysis has been conducted with Cenarchaeum symbiosum, a symbiont of a marine sponge (111), a Pseudomonas-like bacterium that is a symbiont of Paederus beetles (110), Buchnera aphidicola, an obligate symbiont of aphids (1), the Actinobacterium Tropheryma whipplei, the causal agent of the rare chronic infection of the intestinal wall (13, 31), and the Proteobacterium symbiont of the deep sea tube worm Riftia pachyptila (75). These systems provide good models for metagenomic analysis of more complex communities and thus warrant further attention in this review, although the term metagenomics typically connotes the study of multispecies communities. Therefore, the following section focuses on two of these obligate symbionts and the insight into their lifestyles offered by metagenomic analysis. Buchnera-aphid symbiosis. The first genome reconstruction of an uncultured organism was that of Buchnera aphidicola, the endosymbiont of aphids. The relationship between the bacterium and the insect is ancient, leaving each partner unable to function independently of the other, as is reflected in the genomic analysis. Moran's group isolated bacterial DNA from the insect and sequenced and reassembled the bacterial genome. The genus Buchnera contains a "reduced" genome of 564 open reading frames. Upon comparison with a reconstructed ancestral genome, 1,906 genes appear to have been lost. Most of the functions are associated with biosynthetic pathways contributed by the host, suggesting that the genome shrinkage is the result of the symbiotic lifestyle, which has become obligate because of gene loss (1, 42, 101). The reconstruction of B. aphidicola's genome provided insights into the evolution of the symbiosis between the insect and bacterium, the biochemical mutual dependence that they have developed, and the mechanisms of genome shrinkage and rearrangement. The success of genome reconstruction with a single uncultured species provided part of the impetus needed to propose sequencing and reconstructing genomes in more complex assemblages. Proteobacterium-tube worm symbiosis. Riftia pachyptila, the deep sea tube worm, lives 2,600 m below the ocean surface, near the thermal vents that are rich in sulfide and reach temperatures near 400°C. The tube worm does not have a mouth or digestive tract, and therefore it is entirely dependent on its symbiotic bacteria, which provide the worm with food. The bacteria live in the trophosome, a specialized feeding sac inside the worm (32). The bacteria and trophosome constitute more than half of the animal's body mass. The bacteria oxidize hydrogen sulfide, thereby producing the energy required to fix carbon from CO2, providing sugars and amino acids (predominantly as glutamate) that nourish the worm (55, 84). The worm contributes to the symbiosis by collecting hydrogen sulfide, oxygen, and carbon dioxide and transporting them to the bacteria on hemoglobin-like molecules (3, 46, 149-153). The bacterium is a member of the -Proteobacteria, as identified by 16S rRNA gene sequence (47). The bacteria have not been grown in pure culture in laboratory media, but they provide an excellent substrate for metagenomics because they reach high population density in the trophosome and exist there as a single species. Hughes et al. (75) isolated DNA from the bacterial symbiont and constructed fosmid libraries from it that were used to understand the physiology of the bacteria. Robinson et al. (116) identified a gene with similarity to ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) from the same fosmid library. All of the residues associated with the active site are conserved in the protein sequence deduced from the DNA sequence, and it has highest similarity with the RubisCO from Rhodospirillum rubrum. The characterization of this gene lends further support to the premise that the chemoautotrophic bacterial symbiont in R. pachyptila fixes carbon for its host. " Note the picture of the first one clone from a phage! mmbr.asm.org/cgi/content/full/68/4/669/F3the phylogenetic tree of Bacteria, including Archaea! mmbr.asm.org/cgi/content/full/68/4/669/F1Phylogenic strains: mmbr.asm.org/cgi/content/full/68/4/669/F2" APPROACHES TO METAGENOMIC ANALYSIS Top Previous Next References Metagenomic analysis involves isolating DNA from an environmental sample, cloning the DNA into a suitable vector, transforming the clones into a host bacterium, and screening the resulting transformants (Fig. 3). The clones can be screened for phylogenetic markers or "anchors," such as 16S rRNA and recA, or for other conserved genes by hybridization or multiplex PCR (136) or for expression of specific traits, such as enzyme activity or antibiotic production (39, 44, 61, 78, 87, 88, 91, 93, 118, 125), or they can be sequenced randomly (141, 142). Each approach has strengths and limitations; together these approaches have enriched our understanding of the uncultured world, providing insight into groups of prokaryotes that are otherwise entirely unknown." antiobiotics discovered: mmbr.asm.org/cgi/content/full/68/4/669/F4Intracellular screen for quorum sensing inducers: mmbr.asm.org/cgi/content/full/68/4/669/F5FIG. 6. Metagenomics-based model of biogeochemical cycles mediated by prokaryotes in acid mine drainage mmbr.asm.org/content/vol68/issue4/images/large/zmr0040420700006.jpeg"Metagenomics has changed the way microbiologists approach many problems, redefined the concept of a genome, and accelerated the rate of gene discovery. The potential for application of metagenomics to biotechnology seems endless. Functional screens have identified new enzymes (39, 52, 53, 67, 68, 72, 73, 88, 93, 104, 118, 122, 124, 144) and antibiotics (22, 23, 39, 61, 69, 91, 110, 118, 146) and other reagents in libraries from diverse environments. A number of barriers have limited the discovery of new genes that provide insight into microbial community structure and function or that can be used to solve medical, agricultural, or industrial problems. " mmbr.asm.org/cgi/content/full/68/4/669#Modern_Microbiology—a_Pure_Culture_Is_Not_Enough skytroll
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