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20 posts tagged with dehumidifier.
Displaying 1 through 20 of 20. Subscribe:
Are there any dehumidifiers available to the US not made in China?
I'm looking for a home dehumidifier in the 30-70 pint range. Please suggest me a brand made in the USA, or Gemany, etc that I can buy (like from a chain store) or order? [more inside]
Which moisture absorber should I use?
Amazon sells a bunch of different kinds of moisture absorbers, mostly silica based. They are variously "rechargeable," by plugging them in or putting them in the oven for hours (!). Reviews are all over the place. I need to be able to store some mattresses wrapped in plastic in my damp basement. Which absorber(s) do I want for placing inside the plastic?
Dehumidifier with short: fix or dispose of? If so, how?
I have a Frigidaire dehumidifier with a short (when I plug it in, it trips the fuse on any house circuit I try). Should I try to fix it, or is it likely to be unobvious? I'm not particularly handy, but I don't mind taking something apart. Is it safer/better to simply dispose of it? If so, is it recyclable?
I want a QUIET dehumidifier...which won't catch on fire.
The title says it all. EVERY SINGLE dehumidifier I have for my personal use, as well as the ones we use at my workplace's records storage rooms have now been recalled. Has anyone used anything worth buying? I want one which is quiet and will auto-restart/retain settings after a power failure [more inside]
Seeking the Bosch of dehumidifiers. Bonus if it's also the Prius.
Can you help us pick a super-reliable, long-lasting dehumidifier for our damp basement? It would be fantastic if it's also unusually energy-efficient. [more inside]
Where can I buy a dehumidifier today in the San Francisco Bay Area?
Where can I buy a powerful (60-75 pint) dehumidifier in the Bay Area today? I'm willing to buy it online and pick it up in the store, or just go directly to the store, but I'm trying to find a way to avoid spending all afternoon driving from store to store. Is there a specialty shop? Have you had any luck with "buy online - pick up in store today" options at Home Depot, Lowe's, Ace, Target, etc.? Is there a website that compiles local inventory for things like this? [more inside]
Hemp me dehumidify
How can I quickly reduce relative humidity in a small enclosed environment within a fume hood without raising air temperature? [more inside]
Should we get a dehumidifier or do something else?
Is a dehumidifier the right solution for my problem? What else should we be considering? [more inside]
Dehumdifier water is basically distilled water, right?
Is water from my dehumidifier's collection tank safe for tadpoles to live in? [more inside]
Cheap DIY way to dehumidify a basement during a rainy spell?
What is the best creative way to dehumidify a basement without buying an expensive dehumidifier? I don't want to run one constantly, but with all the flooding rain, I need to do something. [more inside]
Moist is a horrible word
My new dehumidifier has me curious...how many gallons of water are in the air in my building at a given temp and relative humidity? Plus a couple of related questions. [more inside]
Good dehumidifier for crawl space?
What's a good dehumidifier for a crawl space? I've read the posts on dehumidifiers for basements, but does a crawl space require something more robust? I'm trying to decide whether I can get away with something I can pick up easily at a big box store for a few hundred dollars, or whether I need to spend double or triple that.
Another Sauna Showdown
[Arachnid Filter] Why don't these two spiders eat each other? [more inside]
DIY dehumidifier?
Looking for a cheap, simple, DIY technique to dehumidify my apartment. [more inside]
Cheap "Dry Box" For Camera Gear?
Are there cheap alternatives for the large and elaborate "dry boxes" I see listed for use as dehumidified storage cabinets for lenses and other camera gear? I've built up a little collection, live where the summer humidity is often high, and do not run my AC 24/7. My first thought is that this is basically an airtight box with a bunch of dessicant inside. Can't really be that simple, though.
My basement smells of decaying fruit. Help!
My basement smells like rotting fruit. What is causing it and, since I'm thinking it needs dehumidifying, any dehumidifier tips or recommendations? [more inside]
Mold and Mildew Prevention
What is the better mold & mildew prophylaxis: a steam cleaner, a dehumidifier, or something else? [more inside]
moldy mouldering mold
Will a dehumidifier work for my mold problem? The size charts say a 25 pint one (~$150) would be fine for my square footage, but would one be able to cover the bathroom, living room and bedroom considering they are separate rooms? Would the air flow be good enough, or should I get some of the much smaller (seems like less well-reviewed too) dehumidifiers that go for $50-$70 on Amazon and put one in each room? [more inside]
Begone, basement humidity!
Are you experienced . . . with buying dehumidifiers? [more inside]
Out damn humidity, out!
Questions about dehumidifiers. [more inside]
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Tarah Donoghue, Class of 2000, presented an outstanding graduation speech at today's Prize Day Ceremony.
Ms. Donoghue earned a double major in Government and Art History from Georgetown University. Her early career was spent at the White House - first as an Assistant Press Secretary to the Vice President and then as Deputy Press Secretary to Laura Bush, the First Lady.
After leaving the White House, Ms. Donoghue was an associate at Gibraltar Associates, a public relations firm. Ms. Donoghue is currently a Communications Consultant with the Susan G. Komen for the Cure Foundation in Boston.
Following are her remarks:
St. Mark's students, faculty and staff, parents and families, members of the Board of Trustees…Good morning and thank you.
I am deeply honored to be with the St. Mark's School class of 2009.
Your class is composed of a diverse and talented group of individuals, representing more than ten countries and twenty-one states across America.
Tomorrow you will be unified by one similarity: You will all be St. Mark's graduates.
Here in Southborough, you have received the guidance and care of a talented and selfless faculty—some who have been your biggest critics…but also your biggest fans.
You have learned the life long lessons of playing team sports—winning with confidence and class… losing with grace and humility.
You have been inspired by the sources of creativity and culture that this school offers—nurturing and supporting your individual interests and passions.
Today I can see the pride in the faces of your parents and families. They have worked hard to give you the unique opportunity of a St. Mark's education, and you will appreciate their generosity more and more with each year of your life.
My days since leaving St. Mark's have been shaped by the values, the knowledge, and the friendships I took away from here.
And I'm grateful that my parents gave me such a tremendous foundation for my life.
In preparation for this speech, I thought back to my own Prize Day in 2000 and tried to remember the words of advice from my graduation speaker…
He was a St. Mark's alum and founder of the financial web site Motley Fool named David Gardner. These days, I wish I had taken notes at David's speech—I might be better off financially…
But what I do remember most vividly that day is the black eye I suffered from a lacrosse game the weekend before. In fact, when I was invited to speak today I was excited at the prospect of re-doing some of those old photos from my Prize Day…
But I know my main responsibility is to share with you a bit about the adventures I have had since graduating from St. Mark's and my hopes for each of you as you leave campus.
This morning I want to talk about opportunity. Opportunity is what I know, and I found so much opportunity because of the skills and spirit of discovery I developed at St. Mark's.
For me, a career path I never expected to pursue arose from seizing a random opportunity. My work in government began from responding to a bulletin board posting in the Georgetown University Government Department, where I saw a flyer seeking White House interns.
So if you think you have a clearly defined path for the next four years, I would encourage you to think again and to keep your eyes open for new and unforeseen experiences.
The Class of 2009 is tasked with resolving some very unique challenges both within your communities and beyond our borders.
Today I urge you to see these challenges as an opportunity.
Because of technology, much of our world is a closely connected place; we cannot ignore the critical and urgent needs of others far away.
With Skype and Facebook, the opportunity to engage in causes anywhere in the world is just a click away. You're online, and your causes are viral. You can meet people with shared passions on every continent.
But take the next step…Find your area of interest and passion and get out in the field.
Continue and expand upon the great work that you have already started…
I know this past year you have tutored at Immaculate Conception School and provided meals at Our Father's Table in Marlborough. You have worked to reduce greenhouse gas emissions on campus with the Green Team. You have held mock electoral debates and election watching parties in dorm common rooms.
Now is a time when a sense of political interest among young people in this country—no matter what party affiliation—is at its highest point ever.
The 2008 Presidential campaigns brought forth a level of engagement in the future of this country that was unprecedented for our generation.
Use this momentum. Don't let age be a barrier to your achievements. Get right out there and be as active as you can be in your area of interest.
I know that hard work will bring boundless opportunities regardless of your age.
At age twenty-seven I've already been to more places than I thought I would by the time I was 50.
And I've been shocked – and humbled – by the amount of responsibility people have been willing to give me. The opportunities are out there. But you have to look for them.
A little luck does not hurt either…but that does not come out of nowhere. One of my favorite sayings was often spoken by Mr. Wiedergott, a former teacher and Athletic Director at St. Mark's. He always said: "Luck is what happens when preparation meets opportunity."
Former President George W. Bush, with his tendencies to invent words, would call this "prep-or-tunity"...I'm not kidding…
I would like to share a brief story that demonstrates how you will use the lasting skills you have learned here at St. Mark's:
The morning started out like any other White House morning for me, with a 5:45 am arrival to my small East Wing office, and a careful read of the day's newspapers with pen in hand.
I then headed over to the West Wing at 7:30 am for my daily meeting with the other White House press secretaries covering the major news items of the day and our strategies for response…As usual, thirty minutes in the press office for this meeting consumed my entire notebook of paper.
That day was a travel day for the First Lady and her staff, and after my meeting, I raced out to the South Lawn to board Mrs. Bush's motorcade to head out to Andrews Air Force Base.
After what was always a very quick ride, we pulled up on the tarmac next to Bright Star—the Air Force call sign for the First Lady's plane—and boarded our flying office.
Right when I sat down across from Mrs. Bush in the cabin, she expressed great concern about a specific news item of the day—the tragic assassination of a female Afghan government official by Taliban militants.
I briefed her further on what I knew, and she asked that I quickly prepare a press statement for her review by the time we were "wheels down" at our destination.
This situation created a tight but typical deadline for me with the short flight, but I was prepared to be fast and accurate—two requirements of my job working for Mrs. Bush.
After having taken very thorough notes at my morning briefing on this same security issue, I was ready to deliver results to my boss.
I must admit…I have to thank my St. Mark's history teacher, Mr. Lyons, entirely for my detailed note-taking and analysis skills.
The preparation and training I received in his class has stayed with me my entire academic and professional career.
I gained such invaluable skills here at St. Mark's, and you too leave with them today.
While I do not have many years of wisdom to share with you, I have been very fortunate to travel to some exotic places and meet some interesting people.
In closing, I'd like to share a few pieces of advice based on my recent adventures:
First, you cannot always pick and choose the experiences you learn from.
I certainly would have preferred for my first time speaking on the record as a White House spokesperson to have been on the subject of national security.
However, the first time I was called upon to respond to press inquiries was when the Vice President shot a man in the face while hunting in Texas. That was my introduction to crisis communications…
Second, make sure to put yourself in a position where you are constantly challenged and constructively criticized.
When I first started working for Mrs. Bush, I received more red pen marks on my briefing papers and press statements than I could ever imagine. Yet I continued to learn from her and refine my writing.
And oh the delight in seeing a paper marked with nothing in red but the word "good" from the First Lady—the White House editor-in-chief.
Third, be flexible and always resourceful.
Your St. Mark's education has taught you to be problem solvers and creative thinkers. In my foreign travels, I was often called upon to make the seemingly impossible happen—like coordinating live TV interviews from places with no electricity, from remote villages in Mali to the beaches of Midway Island.
I learned that oftentimes, the answer is whatever you come up with to get the job done right.
And lastly, be aware of and open to meeting interesting people. You will likely learn the most from those you least expect.
For me, my most memorable White House moments were not from interactions with high level diplomats or policy-makers, but from my daily conversations with the White House residence staff:
James Ramsey is a White House butler who has served for six administrations; Dale Haney is the head groundskeeper who has been there for 36 years; and White House florist Nancy Clarke served since the Carter Administration.
I learned so much from taking the time to get to know each of these individuals who have experienced history from behind the scenes.
For me, getting involved was about serving in our nation's government.
I had the incredible chance to be a temporary custodian of a job where there was a sense of living history around me every day.
It was a great adventure and I hope each of your will leave here today and look for opportunities where you will both challenge yourselves and serve others…and in doing so find adventure too.
Congratulations to all of you. Thank you so much for giving me the chance to share this important day with you.


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MP calls for hormone block drugs for trans children
Post your comment
Comments on this article are now closed.
Reader comments
The awful thing about the existing UK policy is that it flies completely against the balance of evidence and risk and treats the existing "do nothing" approach as though it is the zero risk option against the presumed higher risk of hormone blocking intervention.The opposite is true in fact. Quite apart from the suicide risks highlighted in this article doing nothing involves permitting changes to occur which the patient will then have to deal with for the rest of their lives. By contrast the hormone blocking route does nothing irreversible, has a secure evidence base and buys time for the young person to think and grow in their ability to make fully informed decisions about their own treatment.
As a 30-something post-op transwoman, being able to choose to have my puberty halted would have helped prevent the daily problems I face. It was having to go through puberty "in the wrong gender" that caused me more harm. I welcome the opportunity for transpeople of whatever age to have an easier path, after all we are born into the wrong body, it is not a lifestyle choice.
I agree whole heartadly with Michelle and Christine, the treatment of children with GID is so cruel.I first became aware my body was wrong at 3, yet for various reasons I felt unable to do anything about it until I was 38. I am now with a gender clinic, 11 months + on HRT, and seeing some wonderful changes.If more help, education and access to hormone blockers had been available in the late 70's early 80's, then I would not have suffered the awful masculization effects of male puberty. Which happened late in life for me, aged 17, and my body made up for lost time and my feminine looks evaporated very quickly.I too have contemplated suicide prior to starting my transition process, these children need to be listened to by the medical profession.HBS is not a life choice.So I fully support the campaign backed by Dr Jones.Stephanie
As a transgendered woman, I feel it is incredibly patronising for those not suffering from gender disphoria to have such a controlling impact on the quality of such a young persons future. Thus denying them drugs which will help stabilise their appearance until aged 16 or 18 when they could legally make the descision.
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gets a new look !
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A Short film by Hugo Chesnard
Produced by Butterfly Productions
Production year : 2011
It is about the course of a future expelled person, about the judicial machine whichentails the illegal immigrants to leave the country. This story is taken from real facts arisen during the summer, 2006 in Paris. The story is redrawn under the shape of a social operetta. Souleymane Bagayogo employee, taxpayer without papers, was arrested at his workplace, then expelled towards Mali. A little bit earlier, he had assigned to labor relations board his make his employer respect his rights.
Pierre-François Bernet, Pauline Seigland
Serge Balu, Damien Tronchot, Antoine Larcher


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Someone had the great idea of putting a high-pressure, heat-resistant camera inside the cylinder of a four-stroke engine. Shooting at 1000 fps, the camera recorded just exactly what happens to make engines run. Suck, Squeeze, Bang, Blow! [LiveLeak]
Micro Camera Shows the Workings of a Combustion Engine
Discuss13 Comments | Reply
Ruben GuestDecember 25, 2007 9:08 am
I'm an Engg student and I never really liked ICengines in Mechanics.
by the looks of it, it looks like a Petrol Engine (ifits a spark plug. dunno what a fuel injector looks like, if the white thing is a fuel injector, its a diesel engine). Its 4 stroke.
But this is way cool than how our lecturer says.
Jason GuestDecember 25, 2007 10:05 am
That is a four stroke gasoline engine. The white thing is the spark plug, the mist coming from the valve on the right is the fuel/air mixture (can't tell if it is fuel injected or if it has a carburetor), and the valve on the left is the exhaust valve. This video shows why it is called a four stroke engine... intake (when the intake (right) valve is open and the piston (big thing on the bottom in the middle) is going down creating a vacuum in the cylinder, compression (both valves are closed and the piston is compressing the fuel/air mixture), power (the spark plug ignites the highly flammable fuel/air mixture pushing the piston down with great force), and exhaust (the exhaust valve (left one) opens while the piston once again goes up expelling the burnt gas through the exhaust ports and once the exhaust valve closes the intake valve opens).
This video was helpful in explaining the combustion chamber to my girlfriend.
Dave Bowes GuestDecember 28, 2007 8:06 am
Neat video. However the sharp eyed observer will not that it is imcomplete. It shows the exhaust valve opening but you don't see the piston coming up on the exhaust stroke and the exhaust valve closing. The video jumps from exhaust valve fully open to exhaust valve closed and intake starting to open. So you don't get to see if there is any exhaust - intake overlap.
The compression cycle shows the spark occuring just before TDC implying that the engine is running at low speed.
The video will be the same if the engine has a carb or is fuel injected.
Tom GuestSeptember 30, 2009 12:17 am
Edwin, how can you see flames exiting? you would need a camera on the other side of the valve.
what you are seeing is the petrol that is present on the valve burning off. The condition of the valves cannot be determined from this video.
I would love to see this same video camera technology used in a compression ignition engine.
bruce thompson GuestOctober 1, 2009 11:24 am
We did a bunch of this stuff for Cummins on the 5.9 B series(Dodge pickup app) with a much whiter more solid flame front expanding from the nozzle..THAT flashing off the intake valve appears to be still wetted fuel there causing a richer/visible orange trail. We saw effects like that on bad/dribbly injectors(CI, Diesel)
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Freedom of the Seas Cruise Review by sailingsue1: 5 night get-away
Overall Member Rating
5 night get-away
Destination: Western Caribbean
Embarkation: Tampa
We were happy to know that there was an "adult" only area to relax during this cruise. When we arrived we found out that the cruise lines define adults as 16 years old. The pool and hot tub had kids with glass containers and drunk noisy groups standing around. Security is hard to find and not around to help tame these people.
There was a crying (screaming) baby in the late (8:30) seating and the staff would do nothing. The rules state that age 7 and younger are to use the early sitting. At the customer relations desk, a woman told me that some people have different customs and the baby was allowed. There is a baby sitting service on board but these people chose to not use it. This child was clearly unhappy and it was stressful to see this sadness.
On the positive side, the stateroom attendents are wonderful and bend over backwards to help. The food was good, and the public areas clean.
By the way, the shows are not at all Broadway quality. There was a skilled More and funny juggler there and I hope that he finds his way to a ship that is worthy of his talent. Less
Published 01/02/13
Read more Freedom of the Seas cruise reviews >>
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Cabin review: F3524 Large Oceanview Stateroom
Room too small, closet cramped, lots of counter storage spaceRead All Large Oceanview Stateroom (F) Reviews >>
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Better than Expected
Great Time on Freedom
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Really enjoyable - give it a g...
Great Price - Great Time
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7 Days of Funk video contest: the runners up
- March 04, 2014
Late last year we launched a video contest for "High Wit Me" by Dam-Funk & Snoopzilla, a bonus track from their album 7 Days of Funk. We offered a download of green-screen footage of Snoop & Dam-Funk performing the song, along with these simple rules: "You can do anything you like as long as it's creative - shoot new footage, animation, found footage, Snoop and Dam green screen footage or combinations of all of the above." Snoop & Dam looked through all the submissions and chose their favorite five clips.
THE WINNER: 7 DAYS OF FUNK "HIGH WIT ME" by DANIEL FARAH
First runner-up: Directed by Jen Go.
Second runner-up: directed by Stefloo, aka Staif Floot, who says: "A one-night labor of love, for hip-hop heads, retro gamers and of course weed smokers!!!"
Third runner-up: Directed by Egidio Iovene aka EDG, who says: "Create, eat, listen, love, smoke. God bless Stones Throw for this art & music." Thanks for the video Egidio!
Fourth runner-up: Directed by The Hynie Spanker
TERMS & CONDITIONS OF ENTRY aka THE BLAH BLAH BLAH:
Entrant warrants and represents that the Video: A. is original; B. does not infringe the intellectual property, privacy, publicity, and/or ownership rights of any third party; C. is suitable for public viewing. | By submitting an Entry, Entrant acknowledges that Stones Throw Records is granting a conditional license to use the Green Screen footage of Dam-Funk and Snoopzilla (aka Snoop Dogg) and the Composition and Master Recording "High Wit Me" and Entrant represents that he/she owns all rights to each element of his/her Entry (excluding the Green Screen footage and Composition and Master Recording). | Each Entrant irrevocably grants Stones Throw Records and it's successors and assigns, the unconditional and perpetual right and permission to reproduce, encode, store, copy, transmit, publish, post, broadcast, display, publicly perform, monetize, adapt, exhibit and/or otherwise use or reuse (without limitation as to when or to the number of times used), on an exclusive basis in any and all media without limitation as to time or territory, and without additional compensation or approval from the Entrant or any other party. Entrant waives intellectual property rights, privacy/publicity rights or other legal or moral rights that might preclude the Stones Throw Record's use of any element of the Entry, and agrees not to sue or assert any claim against the Stones Throw Records for the use of any element of the Entry. | Entrant agrees to indemnify and hold Stones Throw Records and its affiliates, officers, directors, agents, co-branders or other partners, and any of their employees (collectively, the "Indemnitees"), harmless from any and all claims, damages, expenses, costs (including reasonable attorneys' fees) and liabilities (including settlements), brought or asserted by any third party against any of the Indemnitees due to or arising out of the use of any element of Entrant's Entry, or the Entrant's conduct in submitting an Entry or creating a video or otherwise in connection with this Contest, including but not limited to claims for trademark infringement, copyright infringement, violation of an individual's right of publicity or right of privacy, or defamation. | Stones Throw Records, in its sole discretion, may accept or deny any Entry to participate in the Contest.
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A specific mix of generalists: bacterial symbionts in Mediterranean Ircinia spp.
Microbial symbionts form abundant and diverse components of marine sponge holobionts, yet the ecological and evolutionary factors that dictate their community structure are unresolved. Here, we characterized the bacterial symbiont communities of three sympatric host species in the genus Ircinia from the NW Mediterranean Sea, using electron microscopy and replicated 16S rRNA gene sequence clone libraries. All Ircinia host species harbored abundant and phylogenetically diverse symbiont consortia, comprised primarily of sequences related to other sponge-derived microorganisms. Community-level analyses of bacterial symbionts revealed host species-specific genetic differentiation and structuring of Ircinia-associated microbiota. Phylogenetic analyses of host sponges showed a close evolutionary relationship between Ircinia fasciculata and Ircinia variabilis, the two host species exhibiting more similar symbiont communities. In addition, several bacterial operational taxonomic units were shared between I. variabilis and Ircinia oros, the two host species inhabiting semi-sciophilous communities in more cryptic benthic habitats, and absent in I. fasciculata, which occurs in exposed, high-irradiance habitats. The generalist nature of individual symbionts and host-specific structure of entire communities suggest that: (1) a 'specific mix of generalists' framework applies to bacterial symbionts in Ircinia hosts and (2) factors specific to each host species contribute to the distinct symbiont mix observed in Ircinia hosts.
- sponge
- bacterial symbionts
- host specificity
- phylogenetics
Sponges are sessile, filter-feeding invertebrates that inhabit diverse marine ecosystems and host remarkably abundant and diverse microbial symbiont populations (Taylor et al., 2007; Webster & Taylor, 2011), in some hosts accounting for up to 35% of sponge biomass (Vacelet, 1975) and consisting of hundreds to thousands of symbiont taxa (Webster et al., 2010; Lee et al., 2011). These diverse symbiont communities may enhance sponge holobiont metabolism through microbial processes, including photosynthesis (Erwin & Thacker, 2008), nitrification (López-Legentil et al., 2010), and sulfate reduction (Hoffmann et al., 2005), and can produce defensive secondary metabolites (Flatt et al., 2005) that decrease the susceptibility of host sponges to predation and fouling (Paul & Ritson-Williams, 2008). In turn, sponge-associated bacteria may benefit from the unique microenvironment within host tissues, potentially nourished by ammonia as the end product of animal metabolism and protected from open-ocean predation and UV exposure. Although empirical evidence for symbiont benefit is scarce (Taylor et al., 2007), the high biodiversity of sponge-associated microorganisms and their exclusivity to host sponges suggest that these niche habitats are fertile grounds for marine microorganisms.
The study of sponge microbiology has revealed striking trends in the distribution and specificity of microbial symbionts, yet little is known about the factors that structure these communities. Evidence for sponge-specific microbial lineages, or symbionts shared among unrelated sponge hosts from distant geographic regions (Hentschel et al., 2002; Olson & McCarthy, 2005; Hill et al., 2006), suggests a generalist distribution of symbionts and some degree of conformity in the sponge microbiota (Taylor et al., 2007). However, from a host perspective, the composition and structure of microbial symbiont communities have been recently reported as species-specific, despite the presence of sponge-specific clusters within these communities (Webster et al., 2010; Lee et al., 2011). Multiple factors may influence the composition and structure of microbial symbionts in sponges, including environmental factors such as temperature and nutrient levels (Mohamed et al., 2008b; Webster et al., 2008; Turque et al., 2010; Webster et al., 2011). In addition, the presence of distinct symbiont communities among unrelated sponge hosts from the same habitat (Lee et al., 2009b; Radwan et al., 2010; Webster et al., 2010; Lee et al., 2011) and similar symbiont communities among related sponges from different oceans (Montalvo & Hill, 2011) suggests that host-specific factors also play a role in structuring the sponge microbiota. Uncoupling the effects of environment and host on symbiont communities requires the study of related sponges from the same environment, a sampling design that to date has rarely been employed (Lee et al., 2009b) or limited to a specific portion of overall symbiont communities (Erpenbeck et al., 2002; Thacker & Starnes, 2003).
To investigate the structuring and specificity of bacterial symbionts in closely related high-microbial-abundance (HMA) host sponges, we studied three sympatric species (i.e. sponges with overlapping species distributions) in the genus Ircinia from the Mediterranean Sea. The genus Ircinia is a chemically diverse and symbiont-rich sponge taxon that exhibits high species richness, occurring in shallow to deepwater habitats of tropical and temperate marine environments. The bacterial diversity of Ircinia spp. is consistent with other HMA host sponges, composed largely of sponge-specific sequences from Acidobacteria, Actinobacteria, Chloroflexi, Nitrospira, Poribacteria, and Proteobacteria (Schmitt et al., 2007, 2008; Mohamed et al., 2008a, b, 2010; Webster et al., 2010; Yang et al., 2011). These studies have focused on the microbiota in Caribbean and Indo-Pacific host species, whereas the molecular diversity of microbial symbionts in Mediterranean Ircinia hosts has been addressed by a single study that focused specifically on cyanobacteria (Usher et al., 2004).
In this study, we characterized the bacterial communities in the common Mediterranean species, Ircinia fasciculata, I. variabilis, and Ircinia oros, using replicated clone libraries of 16S rRNA gene sequences and compared the richness, diversity, structure, and specificity of symbiont communities among these congeneric and sympatric hosts. Phylogenetic analyses were conducted to compare symbionts in Mediterranean Ircinia spp. with previously described sponge-associated bacteria, including sequences derived from Caribbean Ircinia spp. In addition, we resolved the phylogenetic relationships among the three host sponges using ribosomal and mitochondrial DNA markers, thus allowing for the determination of symbiont specificity within a well-defined host phylogenetic context. Herein, we use the terms 'semi-sciophilous' and 'photophilic' to refer to benthic communities commonly inhabited by Ircinia spp. Semi-sciophilous ('shade-loving') communities are found in low-irradiance and cryptic (i.e. less exposed) habitats, such as vertical walls and overhangs, while photophilic ('light-loving') communities occur in high-irradiance and exposed habitats, such as horizontal substrates.
Materials and methods
Sample collection
The marine sponges I. fasciculata (Pallas 1766; Fig. ⇓a), I. variabilis (Schmidt, 1862; Fig. ⇓b), and I. oros (Schmidt, 1864; Fig. ⇓c) were collected from shallow (< 20 m) littoral zones at three neighboring sites (< 12 km apart) along the Catalan Coast (Spain) in the northwestern Mediterranean Sea. Ircinia fasciculata colonies were sampled at Punta de S'Agulla (Blanes; 41°40′54.87″N, 2°49′00.01″E), I. variabilis at Mar Menuda (Tossa de Mar; 41°43′13.62″N, 2°56′26.90″E), and I. oros at Punta Santa Anna (Blanes; 41°40′21.48″N, 2°48′13.55″E) by SCUBA during three consecutive days in March 2010. At each site, ambient seawater samples (500 mL) were collected simultaneously and in close proximity (< 1 m) to sampled sponges. Sponge and seawater samples were transported in an insulated cooler to the laboratory (ca. 2 h transit time), where sponge samples were preserved in 100% ethanol and stored at −20 °C and seawater samples were concentrated on 0.2-μm filters and stored at −80 °C.
Transmission electron microscopy (TEM)
To visualize the bacterial diversity present in I. variabilis, I. fasciculata, and I. oros, small ectosomal and choanosomal tissue pieces (ca. 4 mm3) were dissected and fixed in a solution of 2.5% glutaraldehyde and 2% paraformaldehyde, buffered with filtered seawater. Samples were incubated in the fixative mixture overnight at 4 °C and subsequently rinsed with filtered seawater to remove fixative and then dehydrated in a graded ethanol series and embedded in Spurr resin at room temperature. A Reichert Ultracut microtome was used to produce ultrathin sections (ca. 60 nm) that were contrasted with uranyl acetate and lead citrate for ultrastructural observation (Reynolds, 1963). TEM observations were made at the Microscopy Unit of the Scientific and Technical Services of the University of Barcelona on a JEOL JEM-1010 (Tokyo, Japan) coupled with a Bioscan 972 camera (Gatan, Germany).
DNA extraction
DNA extracts were prepared separately for three individuals of each host sponge species (including ectosomal and choanosomal tissue) and three samples of concentrated seawater (one from each collection site) using the DNeasy® Blood & Tissue kit (Qiagen®). Full-strength and 1 : 10 diluted DNA extracts were used as templates in PCR amplifications.
16S rRNA gene sequence clone libraries
The universal bacterial forward primer 8F (Reysenbach et al., 1994) and reverse primer 1509R (Martínez-Murcia et al., 1995) were used to amplify approximately 1500-bp fragments of bacterial 16S rRNA gene sequences from all sponge and seawater extracts. Total PCR volume was 50 μL, including 10 pmol of each primer, 10 nmol of each dNTP, 1× Reaction Buffer (Ecogen), and five units of BIOTAQ™ polymerase (Ecogen). Thermocycler reaction conditions were an initial denaturing time of 2 min at 94 °C, followed by 30 cycles of 1 min at 94 °C, 0.5 min at 50 °C, and 1.5 min at 72 °C, and a final extension time of 2 min at 72 °C. To minimize PCR amplification biases, a low annealing temperature and low cycle number were used and three separate reactions were conducted for each sample. PCR amplification products were gel-purified and cleaned using the QIAquick Gel Extraction kit (Qiagen®). Triplicate PCR products were combined and quantified using a Qubit™ fluorometer and Quant-iT™ dsDNA Assay kit (Invitrogen™). Purified PCR products (ca. 75 ng) were ligated into plasmids using the pGEM®-T Vector System (Promega).
Individual clones were PCR-screened using vector primers, and clones with approximately 1500-bp inserts were purified and sequenced at Macrogen, Inc. A single sequencing reaction was performed for all clones to recover the 5′-end of 16S rRNA gene sequences using the primer 800R (5′-TAC CAG GGT ATC TAA TCC-3′). Ambiguities on sequencing reaction ends were excluded by trimming sequences at the 5′-end to the highly conserved Escherichia coli site 54 and at the 3′-end to E. coli site 754, yielding sequences ranging from 613 to 725 bp (average length = 683 bp) that were used for diversity calculations and phylogenetic metrics. In addition, bidirectional sequencing reactions with vector primers were performed to recover near full-length 16S rRNA gene sequences (range = 1423–1523 bp; average = 1491 bp) of representative clones (total = 39) for phylogenetic analyses. Raw sequence data were processed in Geneious (Drummond et al., 2010), and low-quality sequencing reads were discarded. Sequences were screened for sequencing anomalies (e.g. chimeras) using Mallard (Ashelford et al., 2006) and a reference 16S rRNA gene sequence from E. coli (GenBank accession no. U00096). Putative sequence anomalies were subsequently confirmed or refuted using Pintail (Ashelford et al., 2005) and two related reference 16S rRNA gene sequences. All confirmed chimeras were removed from the dataset. Quality-checked sequences are archived in GenBank under accession nos. JN655200–JN655511.
Operational taxonomic unit (OTU) assignment and composition
Clone library sequences were ascribed to OTUs calculated at different sequence identity percentages (99%, 97%, 95%, 90%, 85%, and 80%) using the nearest neighbor algorithm, as implemented in the mothur software package (Schloss et al., 2009). The observed OTU richness (Sobs) for each bacterial community was compared across different OTU thresholds, calculated as total (combined sequence data by source) and average values (separated sequence data by samples). All subsequent OTU-based metrics were conducted using an OTU classification at 99% sequence identity. Representative sequences from each 99% OTU were analyzed using a nucleotide–nucleotide blast search (Altschul et al., 1990) to find the most closely related sequence, and using the Ribosomal Database Project II (Cole et al., 2007) sequence classifier to assess taxonomic affiliations.
Diversity, structure, and similarity of bacterial communities
The diversity of recovered bacterial communities in Ircinia spp. and ambient seawater was compared using multiple metrics for OTU richness, dominance, and evenness, calculated in the mothur software package. Richness calculations included observed species richness (99% OTUs), rarefaction analysis, and expected species richness using the Chao1 estimator (SChao1). The effect of increasing sequencing effort on OTU richness was estimated using the Boneh calculation (Boneh et al., 1998). Dominance metrics included Simpson's inverse index (1/D) and the Berger-Parker index (d), and evenness calculations included the Simpson's evenness measure (E1/D) and Smith & Wilson's evenness index (EVAR).
Genetic diversity of symbiont communities was compared among host species and seawater sources using nonparametric tests for homogeneity of molecular variance (homova), as implemented in the mothur software package, and an analysis of molecular variance (amova; Stewart & Excoffier, 1996). These tests provide OTU-independent assessments of genetic variation and differentiation within and among bacterial communities inhabiting Ircinia spp. and seawater. Distances were calculated for amova using the Tajima and Nei algorithm with α = 0.05. Using the arlequin software package, version 3.5 (Excoffier & Lischer, 2010), a hierarchical partitioning of genetic variation was assessed across different levels (among sources, among replicates within sources, and among sequences within replicates), and pairwise variation among sources was computed as FST, with statistical significance based on 1000 permutations. Distributions of unique lineages among bacterial communities were examined using a phylogenetic lineage-sorting test (P-test; Martin, 2002), also referred to as the parsimony test (Schloss, 2008), as implemented in the mothur software package.
To determine the distribution of bacterial OTUs within each community, OTU rank-abundance plots were constructed and compared to a fitted log-series and geometric series distributions by calculating the Kolmogorov–Smirnov test statistic (Dmax, α = 0.05 and 0.01). Community similarity among sources was calculated as Bray–Curtis similarity values and visualized in complete linkage similarity dendrograms using primer v6 (Plymouth Marine Laboratory, UK) computer software. Finally, the integral form of LIBSHUFF (∫-LIBSCHUFF; Schloss et al., 2004) was used to test pairwise differences in bacterial communities from each source, implemented in the mothur software package with significance values based on 100 000 randomizations and adjusted using Bonferroni corrections for multiple pairwise comparisons (Sokal & Rohlf, 1995).
Phylogenetic analysis of bacterial symbionts
Phylogenetic analyses of 16S rRNA gene sequences were conducted to determine the affiliations between sequences recovered from Ircinia spp. herein and previously characterized sponge symbionts. In particular, sequences from recent studies characterizing sponge-associated bacterial communities in Ircinia species from the Caribbean were targeted. Publicly available datasets for Ircinia felix (Schmitt et al., 2007, 2008) and Ircinia strobilina (Mohamed et al., 2008b; Yang et al., 2011) were retrieved from GenBank and grouped into 99% OTUs, following the methods employed for the clone libraries herein. Representative sequences from each OTU for I. felix (n = 102) and I. strobilina (n = 156), top matching sequences from blast searches (n = 189), and near full-length 16S rRNA gene clones (n = 39) and partial 16S rRNA gene clones (n = 87) from this study were aligned to the greengenes reference database (DeSantis et al., 2006) using the mothur software package, with an outgroup sequence from Archaea (Haloarcula vallismortis, GenBank accession no. D50581). Maximum-likelihood (ML) phylogenetic trees were constructed in RAxML (Stamatakis et al., 2005) using the General Time Reversible model with a gamma distribution of variable substitution rates among sites (GTR + G). Data were resampled using 100 bootstrap replicates, and a thorough ML search was conducted to optimize the topology and recover the best-scoring tree. Owing to the variable length of 16S rRNA gene sequences being compared (422–1526 bp), a binary backbone constraint tree was constructed from long (> 1000 bp) sequences and used to restrict topology changes when introducing short (< 1000 bp) sequences into the phylogeny. This method allowed for: (1) accurate reconstruction of deeper nodes, based on the most informative sequences, and (2) precise placement of short 16S gene sequence fragments near terminal nodes, for comparative analysis with previous Ircinia sp. datasets (e.g. excised and sequenced denaturing gradient gel electrophoresis bands; Schmitt et al., 2007).
Molecular identification of host sponges
Sponge samples were identified to species using morphological observations, including gross morphology and fiber characteristics, and ribosomal and mitochondrial molecular markers. A segment of nuclear ribosomal DNA, corresponding to the 3′-end of the 5.8S subunit, the entire second internal transcribed spacer (ITS-2) region, and the 5′-end of the 28S subunit, was PCR-amplified following the method described by Erwin & Thacker (2007). A fragment of the mitochondrial gene cytochrome oxidase I (COI), corresponding to the standard barcoding partition (i.e. 'Folmer' partition) (Folmer et al., 1994; Herbert et al., 2003) and the I3-M11 partition (Erpenbeck et al., 2006), was PCR-amplified using a degenerated version of the universal barcoding forward primer dgLCO1490 (Meyer et al., 2005) and the reverse primer COX1-R1 (Rot et al., 2006). PCR amplicons were gel-purified and cleaned using the QIAquick Gel Extraction kit (Qiagen®) and ligated into plasmids using the pGEM®-T Vector System (Promega). Individual clones were PCR-screened using vector primers and then purified and bidirectionally sequenced at Macrogen, Inc. Consensus sequences were constructed by sponge individual (COI) or by clone (rRNA gene sequences), the latter to account for potential intragenomic variation in ITS-2 copies (Wörheide et al., 2004), and archived in GenBank under accession nos. JN655171–JN655199.
Consensus sequences for rRNA gene sequences were aligned using MAFFT (Katoh et al., 2002) with outgroup sequences from Smenospongia aurea (Dictyoceratida; Thorectidae) (Erwin & Thacker, 2007). Alignment of consensus COI sequences included the congeneric Caribbean species I. strobilina (Erpenbeck et al., 2009) and the outgroup species Hippospongia lachne (Dictyoceratida; Spongiidae) (Lavrov et al., 2008). Pairwise genetic distance matrices (uncorrected p-distance) were constructed using the software package mothur. For rRNA gene sequences, ML phylogenies were constructed using PHYML (Guidon & Gascuel, 2003) and the Hasegawa–Kishino–Yano model with a gamma distribution of variable substitution rates among sites (HKY + G), as suggested by FINDMODEL; data were resampled using 100 bootstrap replicates. Neighbor-joining trees were constructed using Geneious and the HKY model of nucleotide substitution; data were resampled using 1000 bootstrap replicates.
Transmission electron microscopy
Electron microscopy observations of host sponge tissue revealed a high density of bacterial cells (Fig. ⇑). Characteristic of HMA sponges, examined sections were comprised primarily of bacterial cells, with only occasional sponge cells (archaeocytes) and structural elements (spongin and collagen fibers). In I. fasciculata and I. variabilis, ectosomal (peripheral) tissue sections revealed dense populations of 'Candidatus Synechococcus spongiarum' identifiable by their characteristic spiral thylakoid membranes encompassing the perimeter of the cells. Synechococcus spongiarum cells dominated the ectosomal regions of host tissue and were observed to be actively dividing, exhibiting several stages of binary fission (Fig. ⇑d and e). In I. oros, no S. spongiarum symbionts or other cyanobacterial cells were observed in ectosomal tissue, rather a high density of heterotrophic bacteria occurred consisting of multiple bacterial morphotypes, some of which were also showing active cell division (Fig. ⇑g). Sections from deeper tissue regions (choanosome) of I. variabilis revealed the absence of S. spongiarum cells and the proliferation of heterotrophic bacteria cells, many with similar morphotypes to those observed in I. oros ectosomal tissue (Fig. ⇑f).
Bacterial OTUs and sampling coverage
Bacterial 16S rRNA gene sequences recovered from I. fasciculata (n = 77), I. variabilis (n = 80), I. oros (n = 82), and ambient seawater (n = 73) grouped into 124 OTUs, defined by 99% or greater sequence similarity. Grouping sequences according to lower identity thresholds reduced the number of OTUs within each source community; however, the same trend in comparative OTU richness among communities was observed (Supporting Information, Fig. S1). Coverage estimates revealed that sampled sponge-associated bacterial communities represented the majority of expected diversity (76.3% in I. variabilis, 80.5% in I. fasciculata, and 81.7% in I. oros), and doubling the sampling effort conducted herein was predicted to produce few (5–6) new OTUs. In contrast, recovered seawater bacterial communities represented less than half (43.8%) of the total expected diversity, and doubling the sampling effort was predicted to produce 12 new OTUs, indicating that more extensive sampling is required to fully characterize bacterioplankton diversity. Similarly, rarefaction analyses showed the sponge-associated bacterial communities beginning to reach OTU saturation, whereas seawater bacterial communities continued to steadily accumulate new OTUs (Fig. S2).
Composition of bacterial communities
Sponge-associated bacterial communities exhibited high diversity and were comprised of nine phyla, including representatives from four classes of Proteobacteria (Table ⇓). Sequences affiliated with Proteobacteria and Bacteroidetes were recovered from all three Ircinia host species and seawater, whereas Acidobacteria, Nitrospira, Chloroflexi, and Gemmatimonadetes were exclusively found in Ircinia-associated communities. Within the sponge-associated microbiota, Deltaproteobacteria-affiliated sequences comprised an abundant component of microbial communities in all three host species (> 15% total clones), although phylum-level differentiation of symbiont communities among hosts was apparent (Table ⇓). Ambient seawater bacteria represented 10 phyla and included four phyla not detected in sponge-associated communities (Table ⇓).
|Phylum (class)
|I. fasciculata
|I. variabilis
|I. oros
|45.4 (16)
|61.3 (22)
|56.1 (17)
|56.2 (28)
|54.5 (65)
|6.5 (4)
|11.3 (6)
|12.2 (4)
|31.5 (16)
|15.0 (26)
|1.3 (1)
|2.7 (2)
|1.0 (3)
|20.8 (10)
|33.8 (11)
|28.0 (10)
|22.0 (10)
|26.1 (31)
|18.2 (2)
|15.0 (4)
|15.9 (3)
|12.4 (5)
|32.5 (2)
|7.5 (1)
|1.4 (1)
|10.2 (3)
|5.2 (3)
|5.0 (3)
|18.3 (3)
|7.3 (8)
|9.1 (3)
|3.8 (2)
|3.7 (2)
|9.6 (7)
|6.7 (14)
|2.5 (2)
|9.8 (5)
|8.2 (4)
|5.4 (11)
|2.6 (1)
|15.0 (1)
|2.4 (1)
|5.1 (2)
|3.8 (2)
|3.7 (2)
|12.3 (3)
|4.8 (7)
|3.9 (3)
|4.9 (2)
|2.2 (5)
|4.1 (3)
|1.0 (3)
|1.3 (1)
|1.3 (1)
|1.2 (1)
|1.0 (3)
|1.4 (1)
|0.3 (1)
|1.4 (1)
|0.3 (1)
|1.4 (1)
|0.3 (1)
|Uncertain classification
|4.0 (2)
|1.0 (2)
Percentage of total clones is shown by bacterial phyla (classes of Proteobacteria indicated with an asterisk). Values in parenthesis depict the number of 99% OTUs within each lineage.
The vast majority of 16S rRNA gene sequences recovered from I. fasciculata (87.0%), I. variabilis (85.0%), and I. oros (81.0%) matched most closely to other sponge-derived bacterial sequences, generally at high sequence identity levels (≥ 97%; Fig. ⇓). Sequences from Ircinia spp. not associated with sponge-derived clones were most commonly matched to sequences from marine sediment (n = 19), corals (n = 4), and seawater (n = 3). Most sequences obtained from ambient seawater bacteria (98.6%) were closely related to sequence from other bacterioplankton sources, with nearly all sequences (90.3%) matching at very high sequence identity levels (≥ 99%; Fig. ⇓).
Diversity, structure, and similarity of bacterial communities
Seawater bacterioplankton communities were clearly differentiated from the sponge-associated bacterial communities, exhibiting higher OTU richness (observed and expected), lower dominance indices, and higher evenness indices (Table ⇓). Among the three host sponges, bacterial communities in I. variabilis and I. oros exhibited very similar richness and evenness values (Table ⇓). By comparison, the bacterial community in I. fasciculata exhibited lower OTU richness and a less even (more dominant) community structure; however, overlapping confidence intervals for index values were observed among all Ircinia hosts (Table ⇓), indicating similar symbiont diversity across the three host species. Genetic diversity analyses revealed similar trends to OTU-based metrics, with bacterial communities in seawater exhibiting significantly higher genetic diversity compared to those in Ircinia sponges (homova, P < 0.005) and no significant pairwise differences in genetic diversity among sponge-associated communities (P > 0.237; Table ⇓).
|Diversity metric
|I. fasciculata
|I. oros
|I. variabilis
|Observed OTUs Sobs
|Expected OTUs SChao1
|40 (32–64)
|43 (36–65)
|53 (40–91)
|188 (105–394)
|Berger-Parker d
|Simpson Index 1/D
|10.3 (7.1–19.1)
|19.4 (13.6–34.0)
|19.9 (14.0–33.9)
|58.4 (36.0–155.2)
|Smith & Wilson Evar
|Simpson Index E1/D
Lower and upper 95% confidence intervals are shown in parentheses where available.
Rank-abundance analyses revealed the presence of a few, dominant OTUs and numerous, rare OTUs within each clone library, consistent with a log-series distribution of bacterial OTUs (Dmax, P > 0.05). The most dominant OTU in the seawater community accounted for 8.0% of the total community, and singleton OTUs (n = 41) accounted for over 80% of all OTUs. In fact, only five OTUs (9.4%) were recovered more than twice in seawater clone libraries. Dominant OTUs in I. variabilis and I. oros accounted for 15% of each community, with the top three most abundant OTUs accounting for one-third of the total community. In I. fasciculata, a single OTU (corresponding to the cyanobacterium S. spongiarum) accounted for over one-fourth of all recovered clones and the top three bacterial OTUs accounted for nearly half (48.1%) of all recovered clones. Approximately half of the OTUs recovered from I. variabilis (n = 19, 55.9%), I. oros (n = 15, 45.5%), and I. fasciculata (n = 15, 51.7%) were singletons.
Minimal overlap in OTU composition was observed between seawater and sponge-derived sequences, consisting of only two shared OTUs between I. variabilis and seawater that accounted for a small portion of the total clones from each library (2.5% and 5.3%, respectively). Comparisons among the sponge-associated bacterial communities revealed four dominant bacterial OTUs (ca. 25% of each clone library) present in all three host sponge species, with an additional seven OTUs shared between I. fasciculata and I. variabilis and six OTUs shared between I. variabilis and I. oros (Fig. ⇓). No additional bacterial OTUs were shared between I. fasciculata and I. oros. The majority of OTUs recovered for each host species' community consisted of symbionts recovered from a single host species (Fig. ⇓) and represented rare OTUs, commonly appearing once (65.5% of specific OTUs) or twice (25.5%) in clone libraries.
Consistent with patterns of symbiont OTU overlap among host sponges, overall bacterial community similarity values were lowest between I. fasciculata and I. oros (16.7%) and the symbionts in these two hosts differed significantly in community structure (LIBSHUFF, P < 0.001; Table ⇑). The microbiota in I. variabilis exhibited higher similarity to I. fasciculata symbiont communities (36.6%), where no significant difference in symbiont structure was detected (P > 0.639; Table ⇑), than to bacterial symbionts in I. oros (31.1%), where significant differences in community structure were detected (P < 0.05; Table ⇑). Further, distinct phylogenetic lineages of symbionts (P-test, P < 0.001) were observed among all pairwise comparisons between source communities (Table ⇑), and amova revealed significant genetic differentiation among all four sources (P < 0.001; 62.1% of genetic variation) and among replicates within each source (P < 0.001; 37.9% of genetic variation).
Phylogenetic analysis of bacterial symbionts
Phylogenetic analysis revealed that sequences recovered from I. fasciculata, I. variabilis, and I. oros formed 56 monophyletic sequence clusters (Fig. ⇓). Nearly half (48.2%) of these clusters were comprised exclusively of sponge-associated bacterial sequences, with an additional 11 clusters (19.6%) consisting of sponge and coral-associated clones. The remaining 18 clades (32.1%) contained nonsymbiotic representatives, most commonly derived from sediment (n = 8) and seawater bacterioplankton (n = 5). Previously described bacterial sequences from Caribbean Ircinia spp. were present in 21 of the 56 clades (37.5%), with I. felix from Florida (n = 16) and I. strobilina from the Bahamas (n = 12) more commonly presenting related symbionts than I. strobilina from Florida (n = 3). Only 1 of the 56 clades (1.8%) was comprised of sequences exclusively from Ircinia spp. (Fig. ⇓c). Notably, symbiont sequences from the unrelated host species Aplysina aerophoba (Order Verongida) from the Mediterranean, Ancorina alata (Order Astrophorida) from New Zealand, Rhopaloeides odorabile (Order Dictyoceratida, Family Spongiidae) from Australia, and Xestospongia muta (Order Haplosclerida) from the Caribbean were also prevalent in these 56 clusters (32.1%, 19.6%, 14.3%, and 10.7% of clusters, respectively; Fig. ⇓).
Phylogenetic analysis was also used to compare the host specificity of bacteria observed among the three Ircinia spp. with host specificity on a broader scale. The four common bacterial OTUs (i.e. generalist symbionts) present in all three Mediterranean Ircinia spp. were related to sequences derived from unrelated host sponge species, nonsponge (coral) hosts, and environmental (sediment) clones (Fig. ⇑). These generalist symbionts corresponded to one Deltaproteobacterium (IRC001) and three Gammaproteobacteria (IRC006, IRC012, and IRC019), with only IRC006 forming a sequence cluster comprised exclusively of sponge-derived clones. The remaining generalist symbiont OTUs formed sequence clusters with not only sponge-derived clones but also coral-derived (IRC001) and sediment-derived (IRC012 and IRC016) bacteria. Further, even the bacterial OTUs identified as specific to a single species of Ircinia in the clone libraries constructed herein (Fig. ⇑) were closely related to sequences derived from unrelated sponge hosts and environmental samples (Fig. ⇑).
Molecular identification of host sponges
Consensus COI sequences from I. fasciculata (n = 3) and I. variabilis (n = 3) and I. oros (n = 2) individuals yielded a 1213-bp fragment encompassing the standard barcoding region ('Folmer' partition; 676 bp) and the I3-M11 region (537 bp). No intraspecific variation was observed among species, and I. fasciculata and I. variabilis exhibited identical sequences across the entire fragment length (Table ⇓). Ircinia oros was clearly differentiated from I. fasciculata and I. variabilis, with higher variability observed in the Folmer partition of COI (Table ⇓). Interestingly, I. fasciculata and I. variabilis sequences were more closely related to the Caribbean species I. strobilina (0.6% divergence) than the sympatric Mediterranean species I. oros (1.8% divergence).
|I. fasciculata
|I. variabilis
|I. strobilina
|I. oros
|H. lachne
|I. fasciculata
|I. variabilis
|I. strobilina
|I. oros
|H. lachne
Values shown as percentages.
Consensus rRNA gene sequences from I. fasciculata (n = 6), I. variabilis (n = 7) and I. oros (n = 8) clones yielded a 650- to 654-bp fragment encompassing the 3′-end of the 5.8S subunit (50 bp), the entire ITS-2 region (243–248 bp), and the 5′-end of the 28S subunit (356–357 bp). Low levels of intragenomic polymorphisms were observed and variable sites occurred in all rRNA gene subunits and regions as inconsistent point mutations (i.e. occurring at different positions). Partial 28S rRNA gene sequences differentiated I. oros from I. fasciculata (1.16% divergence) and I. variabilis (1.10% divergence) but did not resolve the latter two species. ITS-2 sequences exhibited the highest variability and clearly differentiated I. fasciculata from I. variabilis (2.97% ±0.38 divergence), with variability among species (range = 2.43–3.63%) consistently greater than variability within I. fasciculata (range = 0.40–2.02%) and I. variabilis (range = 0.00–0.00%). Phylogenetic analysis of combined ITS-2 and 28S rRNA gene sequences resolved each species into monophyletic clades with high bootstrap support and showed that I. fasciculata and I. variabilis were more closely related to each other than to I. oros (Fig. S3), similar to the bacterial symbiont communities inhabiting these host species (Fig. ⇓).
The Mediterranean sponges I. fasciculata, I. variabilis, and I. oros were shown to host-dense communities of phylogenetically diverse bacterial symbionts, as occurs in congeneric hosts in the Caribbean and more distantly related HMA host sponges. Bacterial communities associated with Ircinia spp. were clearly differentiated from ambient bacterioplankton communities, in terms of richness, diversity, and composition, and were comprised primarily of sponge-specific symbionts related to bacterial sequences derived from related and unrelated host sponge species. Despite the generalist nature of these symbionts among sponge hosts, phylogenetic metrics revealed that each species of Ircinia harbored a unique bacterial community. These differences in symbiont communities among congeneric and sympatric species suggest that factors specific to each host play a role in structuring bacterial symbiont communities in marine sponges.
Host-specific symbiont structuring is particularly notable as the vast majority of the bacterial symbiont communities found in Mediterranean Ircinia spp. were closely related (> 97% identity) to sequences derived from other sponges, often hosts from different oceans. The observed generalist nature of bacterial symbionts, combined with the host-specific structure of entire symbiont communities, suggests that each Ircinia species harbors a specific mix of sponge generalist symbionts. A similar trend was observed in a meta-analysis of bacterial symbionts in 10 sponge species that revealed significant pairwise differences in symbiont community structure among all hosts, despite the predominance of generalist symbionts comprising each microbiota (Taylor et al., 2007). It is important to note that the widespread distribution of generalist symbionts does not imply that they are ubiquitous components of the sponge microbiota. On the contrary, unrelated and geographically distant hosts often share symbionts that are absent from the microbiota of neighboring sponges with close taxonomic affiliations. In the current study, for example, an Actinobacteria-affiliated symbiont (IRC046) of I. oros matched closely (99%) to a symbiont sequence from the Indo-Pacific sponge Theonella swinhoei yet was absent from the microbiota of I. fasciculata and I. variabilis. As a result, the patchy distribution of generalist sponge symbionts can contribute to the differentiation of bacterial communities among hosts.
The persisting question is which factors dictate the distribution of generalist symbionts and account for the observed dissimilarities in microbial community structure among different sponges. To classify the numerous physical, chemical, and biological conditions that may structure symbiont communities, a framework is presented which divides putative factors into four categories based on their source (environment vs. host related) and prevalence among host sponge species (shared vs. specific; Fig. ⇓). The influence and number of factors in each category depends on the geographic and taxonomic scope of each study. Congeneric and sympatric host species were examined herein; thus, numerous environmental and host-related factors were shared among these species and cannot account for the reported differences in symbiont communities. However, factors specific to different host species may represent a source of differentiation for Ircinia-associated bacterial communities.
Environmental factors specific to distinct habitats (i.e. habitat-specific factors; Fig. ⇑) may play a role in structuring symbiont communities, as the host species investigated exhibit different zonation patterns within littoral benthic landscapes. Ircinia variabilis and I. oros occur preferentially in semi-sciophilous communities, commonly inhabiting vertical relief structures where much of the sponge diversity in these habitats exists. In contrast, I. fasciculata is more prevalent in photophilic communities, characterized by high-irradiance conditions and dense algal growth. Irradiance levels are primary factor in structuring sponge assemblages in the Mediterranean, likely as an indirect consequence of stimulating algal growth and competitive pressures (Uriz et al., 1992). The distribution of host species within the local landscape may thus have ecological consequences for sponge–symbiont interactions and dictate symbiont composition by imposing functional performance pressures. For example, the dense populations of cyanobacteria in I. fasciculata may contribute to host nutrition and growth via the transfer of photosynthetic byproducts (Arillo et al., 1993; Erwin & Thacker, 2008; Freeman & Thacker, 2011), thereby enhancing competitive ability and allowing this species to thrive in algal-dominated habitats.
Host factors specific to each sponge species (i.e. species-specific factors; Fig. ⇑) may also influence the composition of symbiotic bacterial communities in sponges, as suggested by the observed correlation between Ircinia spp. phylogenies and symbiont community similarity. Microbial symbionts maintained by vertical transmission, or direct parent-to-offspring passage, are linked to the evolutionary trajectory of host sponges and thus more similar among hosts sharing a more recent ancestor. Vertical transmission has been observed in several marine sponges (Usher et al., 2001; Ereskovsky et al., 2005; Oren et al., 2005; Enticknap et al., 2006; de Caralt et al., 2007; Sharp et al., 2007; Steger et al., 2008; Lee et al., 2009a; Webster et al., 2010), including Ircinia species where successive generations are seeded as larvae with diverse bacterial symbionts (Schmitt et al., 2007, 2008). Therefore, while periodic horizontal transmission of microbial symbionts (i.e. environmental acquisition) may contribute to the homogenization of symbiont communities in unrelated sponge hosts (Schmitt et al., 2008), recurrent vertical transmission may dictate symbiont composition over shorter evolutionary time scales owing to legacy effects of the microbial inheritance. Notably, environmental and host-related factors are not mutually exclusive and may act in concert to structure bacterial symbiont communities in marine sponges.
Discerning the contribution of host ancestry to symbiont structure requires the accurate resolution of evolutionary relationships among sponges based on molecular data and phylogenetic analyses. The current study showed that COI mtDNA sequences (including extended coverage of the I3-M11 partition) were unable to differentiate I. fasciculata from I. variabilis. The utility of the I3-M11 region in differentiating related sponge species (Reveillaud et al., 2011) and as a population genetics marker for intraspecific differentiation in sponges (López-Legentil & Pawlik, 2009; Xavier et al., 2010) suggests that this gene region is either highly conserved within the genus Ircinia or that I. fasciculata and I. variabilis are closely related species. Indeed, some degree of controversy surrounds the taxonomic status of I. fasciculata and I. variabilis (Pronzato et al., 2004), and these species can be difficult to differentiate in the field (Maldonado et al., 2010). Analysis of the second internal transcribed spacer region (ITS-2) of the nuclear ribosomal operon revealed consistent genetic differentiation between I. fasciculata and I. variabilis, indicating that some molecular markers are able to delineate these species. The variable resolution of different genetic markers among sponge taxa indicates that multiple target genes are required to accurately resolve the relationships among species and determine how these relationships may affect the community structure of bacterial symbionts in sponges.
Based on the sponge-specific nature of individual symbionts and host-specific structure of symbiont communities, our results reveal a complex picture of host–symbiont specificity in HMA sponges and suggest that a 'specific mix of generalists' framework characterizes bacterial communities in Ircinia spp. from the Mediterranean. Further, by comparing symbiont communities in congeneric sponges from the same environment, our results suggest that factors specific to the each sponge species may structure the distinct symbiont mix observed in Ircinia hosts, including habitat-specific conditions (e.g. irradiance) and host evolutionary history. Additional research on the spatiotemporal dynamics of bacterial symbionts in sponges and controlled experimental manipulations of sponge holobionts are required to further unravel the multiple and potentially interactive factors that structure the complex sponge microbiota.
Supporting Information
Additional Supporting Information may be found in the online version of the article:
Fig. S1. Diversity (OTU richness) of microbial communities in Ircinia spp. and seawater at different sequence identity thresholds, showing combined data (top) and average values (bottom; n = 3).
Fig. S2. Rarefaction analysis of bacterial community diversity in sponges Ircinia fasciculata, I. variabilis and I. oros and ambient seawater.
Fig. S3. Detailed phylogenetic tree of host sponges (Ircinia spp.).
Please note: Wiley-Blackwell are not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article.
We thank Dr. M. Uriz (CEAB) for help with sponge taxonomy and sample identification and F. Crespo (CEAB) for field assistance. This research was supported by the Spanish Government projects CTM2010-17755 and CTM2010-22218 and by the US National Science Foundation under grant 0853089.
Editor: Max Häggblom
- © 2011 Federation of European Microbiological Societies


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Tag: PalFest
Rahat Kurd: Out of It: An Exchange with Palestinian-British Writer Selma DabbaghOctober 2015
Selma Dabbagh talks about the personal and political in her debut novel, Out of It.
Life Under LockdownAugust 2012
Residents of the Gaza Strip are restricted in their movements, in what they can bring into and send out of their land, even how far off their shores they can fish. Words, though, know no borders.


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Do you use the "No Wait Rebate" option?
I ordered a cpu from tigerdirect.com, which had a $50 MIR. The rebate is processed through Worldwide Rebates, and offered a "No Wait Rebate" option.
Basically, you pay them a small percentage of the rebate (in my case $4, or 8% of the rebate...im not sure if it varies with price), and they "expedite" rebate process from around 5-8 weeks down to 1 week.
So how many people use that option, vs waiting for the entire amount?
4 Comments
Sign up for a Slickdeals account to remove this ad.
can i see the money within 2 weeks?


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Join Couchsurfing to see Yuliya's full profile. It's free!
About Me
we should act more, think less, and stop watching ourselves live (c)
I am not a person of any/many virtues.. someone just have to be simple and usual here.
I do not have enough wit to have a good sense of humor, or to carry a deep conversation.
I am problematic (for myself) and strange (introversive character) which comes from my sensitivity. I do not want to deceive anyone there about me being cool and easy going.
I am very kind (you can not notice this but this is usually what i feel).
I argue.
I have to write something good about myself here:
I am never frightened with your disadvantages and I just can love you with all your bright sides and nightsides. It is like having Stockholm syndrome to everyone.
Why I'm on Couchsurfing
Oh God, some people are just irreplaceable.. Even those one whom I do not keep in touch with any more and they are not on my friends list there, they are still in my heart and I remember them..
Drawing spirals on the snow coasts with the snowboard, reading, dreaming, phisical training, running, traveling, history learning.
Dream - Yachtmaster Sail Training Course:) Mmmm...
- running
- reading
- traveling
- drawing
- sailing
- snowboarding
- history
Music, Movies, and Books
From the last books the one that impressed me - Doctor Zhivago, Boris Pasternak.
One Amazing Thing I've Done
I was travelling in a night train from Pescara to Venice. (In Ukraine in all night trains, there is a porter who hands out linen and 'beds' are separated). There was no porter and I found out a free compartment and beded down on seats that have to be drawn together to create a bed. I was very impressed. My impression grown up more when a yong man joined my appartment and lied down, his knees touched my knees!
Countries I've Visited
Austria, Bulgaria, Croatia, Czech Republic, France, Georgia, Germany, Greece, Hungary, Italy, Netherlands, Norway, Portugal, Russian Federation, Slovakia, Slovenia, Spain, Thailand, Turkey, United Kingdom
Countries I've Lived In
Austria, Ukraine


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Thursday, November 6, 2008
Home on the Range November Highlights
I'll be blogging a lot on the road in November. I'm doing a class for the Academy and then have a few short hops here and there to testify and do some follow up work on some old cases. The posts will be posting, thanks to my own little laptop.
9 comments:
Just don't get that zip caught up in the forward assist. Interesting things might happen.ReplyDelete
OK. I'm looking at it on a Blackberry and when Peter said "forward assist" I went back and gave it a closer look.
There's cleavage in that picture!
And I missed it the first time. Wow, that's not like me; I'm a man! But I'm an old man.
Con sarn it! By cracky! I feel like Gabby Hayes' older brother, Purple.
Put me down for a subscription!ReplyDelete
word: fultust
Glad it wasn't a hot .308 casing.ReplyDelete
I was blistered once when a hot one landed in the pocket behind the right collarbone. I couldn't fish it out fast enough because of my shirt and some webbing. Like most ole honkies of Scotch-Irish descent, it doesn't take much to raise a blister.
I did keep the muzzle pointed downrange.
Somerled, BTDT from a hot 30.06 brass last year at a high-power match. Down the collar, down my back to stop at the belt line between me and my undershirt.ReplyDelete
Big time boogie!!
*whistles, leers, á la Heinlein's The Moon Is A Harsh Mistress*ReplyDelete
- anonnemoNovember 2, 2008 at 3:54 PM
love it.ReplyDelete
Pity, This doesn't seem to be on the news stands in Chicago.ReplyDelete
I started this blog so the child I gave up for adoption could get to know me, and in turn, her children, as well as share stories for a family that lives too far away. So please keep it friendly and kid safe. Posts that are only a link or include an ad for an unknown business automatically to to SPAM..


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"Well it's just unfortunate for him. Especially for a senior and a kid that's done so much for the program. First of all, you hate it for him. You hate it for a team. You hate it for all of Carolina because Bryn's done so much. He's a leader on our football team. He's thrown for over 8,000 yards. He's thrown for 64 touchdown passes. He's done some incredible things.
"We talk about it all the time with our players. You're one play away from it all being over with. So you hope you don't take things for granted. Unfortunately for him it is here at Carolina. We're going to move forward. Marquise (Williams) is going to step up and lead this football team and we're going to move from there."
How has Renner taken the news and how has the team responded?
"Well it's been rough on Bryn. I mean it's been really rough. When the MRI was read the doctor didn't give him any choice at all on the matter. It was we're going to have to have surgery. We're going to need to do it this week and this is it. It's been kind of a shock to him. It really has. He knew it dislocated in the game but he came back in and played. He felt like he would rehab a little bit, get a little bit of strength back into it and be ready to play. He just did not know the extent of the injury at that time.
"Team-wise, we've never mentioned it. We didn't talk about it in meetings. We didn't talk about it at all. We don't ever talk about injuries. We don't talk about them with anybody. Everybody understands in this game that they come and they go. The next guy's got to step up and that's why the next guy's got to be prepared. I think Marquise has done a nice job with that."
How does the rest of your quarterback depth chart look behind Marquise Williams?
"Well I see the next guy that's thrown a touch down pass would be Ryan Switzer so we'd probably put Switzer out there. Quinshad Davis, we'd throw him in there and then we'll just see. No, but Kanler Coker and Mitch Trubisky are preparing right now to be that next guy. We'll just see what happens from there."
Is the goal to keep Mitch Trubisky's red-shirt status for the season?
"Ultimately, yes. But you know if duty calls he's ready to go. I can assure you. He came by the other night when he heard about it. It was about 9:30 at night and he came by the office and said, 'Coach, I just wanted to let you know I'm ready with whatever you need me to do.' He's got the right attitude about it. Kanler's got the right attitude. They all do. Bryn has done a really good job leading that group. He's been the leader in that quarterback room and he's done a good job preparing those kids."
Does it give you more confidence having seen Marquise Williams produce from the pocket already this season as he moves forward as the starter?
"We're very comfortable with Marquise in the game. I think the team is too. That's the silver lining in him playing up to this point is that the team is very comfortable with him. They have confidence in him. They know he can get the job done and they feel good about it. It's not like that everybody's got to rally around a guy that hasn't played where we're all going to have to play super human to be able to get it done. It's not going to need that. Everybody just needs to do their job and he's do his and we'll be okay."
Comments on Landon Turner's play on the offensive line against N.C. State:
"He really did a nice job. We talked about that was the strength of their defense was their four down guys especially inside. Landon, time after time, he dominated his guy. He drove him off the ball in the running game. It was big factor in why we were successful running the football."
Will there be fewer designed run plays for Marquise Williams now that he is the official starter and primarily taking the reps in the pocket?
"We talked about that. Some of the times on the hits, he was just extending plays and doing those things. We're going to always have a couple of quarterback runs that are designed for him because we want to take advantage of what he can do. But yeah, we don't want to get him beat up. No doubt about it."
Comments of Kanler Coker and what he brings to the team from the quarterback position:
"Well Kanler can throw the ball and he can run. He's a dual threat guy. He can do both. If we get to Kanler we feel pretty confident that he will be able to go out there and lead this football team."
Thoughts on the Virginia Cavaliers:
"Well first of all, if you remember last year they lost six games in a row and then they went to N.C. State for their homecoming and won the game. They've been in this situation before. Let me tell you, it's tough when you look at them on film they don't look like a 2-7 football team. They're in every one of these games. You keep looking and you say when's the other team scoring and when's it getting out of hand. It doesn't until late in the fourth quarter. They're playing a lot of young kids. They've got some talented guys. They're doing some good things. We are going to have to play a really good football game."
What have you seen from Jon Tenuta's Virginia defense:
"He's blitzing about 60 percent of the time. He's going to bring the heat and he may even decide to do more now and test Marquise out. We've got to prepare for that. You have to understand that sometimes they're going to get you on some of the blitzes. Hopefully you're going to bust them your share of the time."
Comments on Virginia quarterback David Watford:
"He's a dual-threat guy. He can run and they're taking advantage of him. He's got a nice arm on him. He throws the ball well. They've got a running back that's doing really well and they're trying to get the ball to the five-star recruit, Taquan Mizzell, and they're doing that out in open spaces. It's definitely tough on you."
"Larry Fedora Live" airs weekly during the season on Tar Heel Sports Network affiliates, including UNC's flagship station (WCHL 1360AM/97.9FM).
Follow @insidecarolina


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Friday night was the last day of VBS and something was a little different. Maybe it was the fact that Emma had told everyone in Abby's class that we're adopting, but one of the volunteers that's there every year said that she had also adopted from China. She gave us her contact info and asked us to contact her so that she can get us hooked up with the church's adoption support group and so that the church can help us fundraise. Apparently the church has a heart for adoptions and wants to help in any way possible. That meant the world to me. To have people along this journey that can support us through friendship is priceless. This is an emotional journey that can at times take a lot out of you mentally, physically, and spiritually.
Since we've announced our adoption we've had an overwhelming amount of support, except from our family members. My dad, my cousin, and my aunt are the only family members that congratulated us and been an encouragement. Everyone else has acted as if we're committing some sort of selfish act and seems disappointed in our decision to adopt a child that was abandoned and provide him with a loving home. But I have to remind myself that the people that are not supportive are those that have chosen to not be active participants in our lives, and that although I respect their opinions, I do not need to allow the opinions of others to change whether I obey God.
Although I know that the only thing that matters is that we follow God's guidance in all of this, I still woke up on Sunday morning feeling down by some of the comments I received from family members (some I haven't seen in over 20 years and have never met my kids) over the weekend. I got everyone ready for church and even though we were 10 minutes late walking into a new church, we did it without any issues lol. We decided to visit the church where the girls attend VBS. Emma stayed in the special needs class to hang out with Abby (I love that she wants to be with her), and Lincoln went to the nursery to hang with the other babies.
Matt and I finally made it to the sanctuary and got our seats. The church had a guest speaker and I can't say enough for the amazing word that God gave this man. It was everything I needed to hear. After the service was over we walked out to get the kids and head off to lunch. As we were loading everyone into the car (the 30 minute ordeal that happens when we load and unload lol) one of the kids from Abby's class started talking to Matt from inside his parents car, which sparked a conversation between Matt and the child's parents. Once all the kids were in the car I walked over and we all got to talking. Come to find out, their son Landon (Abby's been in class with him for 3 years) has hydrocephalus and get this...they're also adopting from China and are working with the same adoption agency. They have four biological children and also have the struggles that come with having a special needs child. It was just so absolutely encouraging to meet a family that is in the same shoes medically and is also pursuing adoption. It just confirmed in my heart that it does not matter whether the naysayers accept this as God's will for our lives, because ultimately the only thing that matters is whether we're listening to God, following His lead, and when it comes down to it God knows our hearts. I'm so excited for this new friendship and traveling the adoption and special needs road with our new friends. I can't wait to see God's will unfold in all our lives.
I do want to make sure that I take this time to thank those of you that have been a source of encouragement, we truly appreciate your kind words and prayers. We also want to thank those that have donated to our puzzle fundraiser. We have a $500 fee coming up that we have to send in and the money raised so far is being put towards that. From the bottom of our hearts we're truly thankful for every prayer, loving words, thoughts, and donations.
Thank you so much!!!!!
He predestined us for adoption as sons through Jesus Christ, according to the purpose of His will.
Ephesians 1:5


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