The current state of S. gigantea - acclimation and antibiotic treatment

[D-Nak]

I wanted to start a thread where we can pull our notes together regarding the recent surge in gigantea purchases, but also the increase in deaths associated with those purchases. We need to know the protocol used -- what meds, how long, type of environment, including tank size, whether or not a heater was used, etc. Hopefully we can develop a new protocol to try on future acquisitions. I would like to keep this thread specific to S. gigantea only, as I believe that all anemones species vary slightly, and if we are indeed talking about a bacterial infection, these bacteria are species specific.

My thoughts:
Cipro and more recently Septra have, in the recent past -- up to the last six months of so -- aided in the acclimation process of S. gigantea, saving countless anemones from dying. The assumption is that without antibiotic treatment, the anemone would've died.

Most, if not all, recently acquired -- specifically newly imported -- gigs have died even with the use of Cipro and/or Septra. Several hypotheses have been proposed, but the leading hypothesis is that antibiotics have been added somewhere in the supply chain, causing resistant strain(s) of bacteria.

My understanding is that antibiotics have actually been used for quite some time, the most common being Nitrofurazone, which is useful in koi for the treatment of furunculosis. Assuming the gigantea are exposed to antibiotics, and that the bacteria have now become resistant, we need to determine a protocol to successfully kill the bacteria without killing the zooxanthellae population or the anemone itself.

It's also worth noting that coral bleaching has been studied in great detail for many years, and is applicable to anemones because both contain zoox. Aside from environmental causes, one of the causes for bleaching is Vibrio spp. Unfortunately, theres are hundreds of species of Vibrio, and determining which species is infecting S. gigantea -- if any -- would have to be carried out by the scientific community. This is, of course, assuming it is Vibrio.

My questions:

1. Typical treatment for Vibrio is antibiotic therapy (doxycycline or a quinolone). Being that Cipro is a quinolone, have others tried other quinolones, a combination, or simply doxycycline by itself? I've read many posts about its recommended use, just need to know if anyone has tried it.

2. What are the environmental dangers of the use of other quinolones? We know that Cipro breaks down in light, making it a low impact antibiotic. What about others?

3. Has anyone found a research paper discussing the effects of antibiotics on a zoox population? Can it weaken or even kill zoox?

4. Based on my limited amount of research, Vibro seem to favorably respond to higher water temperatures. What would happen if we lowered the temperature in our QT tanks? What is the lowest temperature gigantea can endure without without causing undue stress?

The future:
I'm wondering if we might want to start experimenting with other types of antibiotics, or a combination of antibiotics. As I am not part of the scientific or healthcare-related communities, I cannot lead the charge. I'd be happy to assist in any way possible, aside from lending a healthy gigantea for scientific research (hah!). Again, the goal of this thread is to start a dialogue to help us move forward, I don't want this thread to dread in the past or lay blame on a particular retailer or wholesaler or exporter. We can't stop what they're doing, but we can develop an effective response protocol based on what we assume they are doing. We just need to work together and be prepared for a long journey.

[D-Nak]

I also wanted to note the following: it was stated on another thread that one of the online dealers has their own treatment protocol. Here it is:

"Prior to introduction into this specialized system, we medicate every single Gigantea, Magnifica, Haddoni, and Mertensii in Ciprofloxacin for seven consecutive days, at a rate of 250mg/10 gallons with a 100% water change performed every morning. I personally have found this regiment to be the most effective. The medication starts immediately upon arrival into our facility following proper acclimation from overseas exporters, importers, or other suppliers we deal with here in the US. After the 7-day treatment, we house the treated anemones in this dedicated system and continue to quarantine and condition prior to offering them for sale. If any anemone out of the batch is not in optimum health, we start all over again and continue with the protocol at a higher dosage. If the anemone(s) pull through, perfect, if not they are unfortunately destroyed, or perish on their own in different holding tank(s). We typically hold all of the above mentioned anemones for a minimum of 3 weeks total, oftentimes much longer as I want to only offer them if they are in optimum health."

If I'm reading it correctly they have the nems isolated for 7 days undergoing Cipro treatment. After the 7 days, if they appear healthy, they move them into the dedicated system.

What we tell people on the boards is to keep the nem isolated for another 7 days without meds for observation purposes. This gives the nem time to either 1)get sick again if not fully cured or 2) allow them to purge the remaining bacteria so they're not "carriers". What the retailer is doing is going straight from QT into the dedicated system without a period of observation (without antibiotics). This could feasibly create a scenario where what appear to be healthy gigs are infecting other gigs, but no symptoms appear until they arrive at our homes. As I said many times, it can take up to a month for symptoms to become externalized where we can see them. Unfortunately, all of us -- myself included -- have rushed this process and we pay with the death of the nem (this is assuming a bacteria is the cause of death).

The reason I mention this is because it's been proven that Vibrio spp. that infect coral can survive in the water column and move to another host. If we are dealing with Vibrio infecting gigantea, they can easily move from tank to tank especially because nems are essentially balloons full of water. It's been documented many times where sick gigs have gotten other gigs sick.

[addictedreefer]

Just to remind everyone that treatment is not always necessary, I have a blue gig going on 4 months that was never treated (despite several deflations early on).

[D-Nak]

Just to remind everyone that treatment is not always necessary, I have a blue gig going on 4 months that was never treated (despite several deflations early on).

I agree. There have been quite a few that didn't need any treatment at all. I have a tiny gig that needed no treatment, while the three others I received at the same time all died, even with treatment.

[tchi]

"at a rate of 250mg/10 gallons with a 100% water change performed every morning"

This may be the reason why the recent DD gigs haven't been responding to treatment. They do a water change in the morning and add the medication when the light cycle begins. While they have good intentions, they fail to realize that cipro has been shown to degrade rapidly under light which is why in Minh's protocol it says to add it after the lights are out so the anemone has maximum contact with the antibiotic throughout the day. If cipro is added in the morning, it is rapidly degrading throughout the morning/afternoon, dramatically lowering the concentration as the day progresses, and essentially turning it from a low concentration bath to almost nothing by the time lights are out. This is the opposite of what the recommended treatment protocol says to do...

[Reefahholic]

I agree D-Nak, we need to develope a new treatment protocol. Not saying the current protocol isn't working, but I believe we can tweak it and make it better.

I've been working on the side of nurses who use multiple antibiotics and see the negatives and positives first hand. My sister is also a nurse. I've got a list of new antibiotics I've been testing the last few weeks. There's a few that have done well, but it's too early for me to see a pattern. I need more time.

Another issue we need to realize is that these infections can be both gram negative & positive. The bacteria can also have a somewhat broad classification as anaerobic, aerobic, or facultative. This is based on the types of reactions they utilize to generate energy for their growth and other activities.

Needless to say, we have a lot more to learn. I hope that in the near future we can save more Gigantea's.

[Breadman03]

"at a rate of 250mg/10 gallons with a 100% water change performed every morning"

This may be the reason why the recent DD gigs haven't been responding to treatment. They do a water change in the morning and add the medication when the light cycle begins. While they have good intentions, they fail to realize that cipro has been shown to degrade rapidly under light which is why in Minh's protocol it says to add it after the lights are out so the anemone has maximum contact with the antibiotic throughout the day. If cipro is added in the morning, it is rapidly degrading throughout the morning/afternoon, dramatically lowering the concentration as the day progresses, and essentially turning it from a low concentration bath to almost nothing by the time lights are out. This is the opposite of what the recommended treatment protocol says to do...

You might have a point there.

[Sugar Magnolia]

I can admit up front that I have no experience treating these nems with antibiotics. My question is, is it or is it not hit or miss treating these nems prophalactically with antibiotics when there is no known diagnosis for what is ailing the anemone? Seems like putting a bandaid on a wound of unknown origin to me. Equating treating an anemone with antibiotics to treating a human with antibiotics seems like quite a stretch to me.

:shrug:

[DasCamel]

I can admit up front that I have no experience treating these nems with antibiotics. My question is, is it or is it not hit or miss treating these nems prophalactically with antibiotics when there is no known diagnosis for what is ailing the anemone? Seems like putting a bandaid on a wound of unknown origin to me. Equating treating an anemone with antibiotics to treating a human with antibiotics seems like quite a stretch to me.

:shrug:

If we practiced this way with humans it would be frowned upon. Well some physicians do,but not the point here. Unfortunately we do not yet have a proper diagnostic tool set to properly ID what exactly is causing the Anemone in question to be sick. Until we do, the antibiotics do seem to be effective in increasing the survival rate.

[OrionN]

About Cipro been degraded by light, I am not sure how fast these are been destroy by light. How fast depends on how much light. If we dose fully and change water 100%, I don't think the degradation of antibiotic mater too much.
Personally, I also find that daily 100% water change, with additional water change as needed works best. I change water whenever there are particles in the water I cannot suck out (the water is cloudy). Lately I used 500 mg Cipro per 10 gal.

Other antibiotics can be use inclueded trimethoprim/sulfamethxazole at the dose of 160/800 mg per 10 gal. Nitrofurantoin at dose of 100 mg per 10 gal. I would still use 100% water change daily with these two medication. Likely Nitrofurantoin would have more narrow spectrum of activity than either ciprofloxacin or Trim/Sulfa.

My last Blue Gigantea from DD, I treated with Trim/Sulfa. He relaped when I have to end treatment early. I put him back in TT and treat him with combination of Ciprofloxacin and Trim/sulfa. He is doing better despite a heat controller mal-function that got the tank temp up to 90 degree for a few hrs (in my QT reef system where I put this anemone in after treatment). Enough to crash the Xenia population I have there but I fixed that with 3 100% water change. The Gigantea seem to have a minor set back but doing well. I will update the thread later.

[OrionN]

I can admit up front that I have no experience treating these nems with antibiotics. My question is, is it or is it not hit or miss treating these nems prophalactically with antibiotics when there is no known diagnosis for what is ailing the anemone? Seems like putting a bandaid on a wound of unknown origin to me. Equating treating an anemone with antibiotics to treating a human with antibiotics seems like quite a stretch to me.

:shrug:

IMO, there is clear cut survival improvement for anemones with antibiotic treatment. The earlier, the better for survival. While it is nice to know exactly what we are treating, Dr. empirically treating without culture result all the time. It certainly cost a lot more to culture everything, and withold treatmetn until we have all the result would kill a lot of sick patients.
I would love to see retailer like DFS culture the next 10-100 sick anemones (what ever number that is feasible for them) and publish the result as to what bacterial species involved. There are a huge number of bacterial species but only a few handful are pathogen (for human). I am fairly sure it is going to be similar in anemone infections.

[TinManx]

D-Nak, Do you currently have any sick giganteas? I still want to try and use aiptasia as a model organism for treatment development. It's documented that pathogenic vibrio sp. can be transfected from sick coral to aiptasia.

[D-Nak]

D-Nak, Do you currently have any sick giganteas? I still want to try and use aiptasia as a model organism for treatment development. It's documented that pathogenic vibrio sp. can be transfected from sick coral to aiptasia.

I don't have any sick gigs right now. I do have a lot of healthy majano in my DT however, you're welcome to have those! :D

[OrionN]

Can't use Aptasia or Majano as model for treatment because they are not the same, or any where near delicate as the anemones we want to keep. Use them as test animal, then everything will work, meaning does not mater what med we do use, they will be fine, even without treatment.

[TinManx]

Can't use Aptasia or Majano as model for treatment because they are not the same, or any where near delicate as the anemones we want to keep. Use them as test animal, then everything will work, meaning does not mater what med we do use, they will be fine, even without treatment.

I am not exactly sure what you are saying. If you are saying that aiptasia are not susceptible to bacterial infection, or that they are not sucseptible to tranfection with vibrio sp. from other species of host, you are wrong. It is published.

If you are saying that treatment developed on a hardy species such as aiptasia may not transfer to S. gigantea due to its perceived delicateness, you may be correct. If you are saying that I may not be able to infect an aiptasia with gigantea disease you may also be correct. But science is about trying things that may not work.

[OrionN]

If we inject vibrio into the aptasia, sure they will get infected. How is that going to be our model for infection of Gigantea or Magnifica? Aptasia will live through anything. You scape them off of the rock with just a little tissue behind and a new on pop up.

If we are going to try to get some useful information regarding infection in host anemones, we will need to have a large number of anemones/sick anemones available. We will need resource to swab and culture these sick anemones and tabulate what are the bacteral species that infect these anemones. The culture also will have antibiotic sensitivity. Once we have these results, it will be easy to determine what is the best antibiotic to try to cover these infection emperically in the future.

Model animal is not needed to gather these type of informations.

[OrionN]

In this issue, we are (or at least I am) almost certaint that most of the time what kill these anemones are bacterial infections. These infections are due to the stress, injury and poor conditions (incompair to ocean condition) in which they are keep after collection. What we need is to improve the condition as much as we can (which have been done I am sure) and ID these pathogens and determine the antibiotic sensitivety of these pathogen so we can use this information and treat them when they are sick.

[TinManx]

If we inject vibrio into the aptasia, sure they will get infected. How is that going to be our model for infection of Gigantea or Magnifica? Aptasia will live through anything. You scape them off of the rock with just a little tissue behind and a new on pop up.

Will they live through vibrio infection. The literature examples say no.

If we are going to try to get some useful information regarding infection in host anemones, we will need to have a large number of anemones/sick anemones available.

To collect the number of gigantea needed for this study would be both inhumane and cost prohibitive (at least on my wallet).

We will need resource to swab and culture these sick anemones and tabulate what are the bacteral species that infect these anemones. The culture also will have antibiotic sensitivity. Once we have these results, it will be easy to determine what is the best antibiotic to try to cover these infection emperically in the future.

What kills cells on a petri dish may not work as a treatment in a QT setting, or may just kill the anemones. Aiptasia are more like gigantea than culture media.

Model animal is not needed to gather these type of informations.

I am confused on this one. How do you think therapeutics are developed?

[OrionN]

Will they live through vibrio infection. The literature examples say no.
I am not sure why injection Vibrio into aptasia would give us any useful information. If I want to kill an aptasia, I just inject kalk into it :). What to do with tiny or inaccesable aptisia is the problem. I just get a CBB.

To collect the number of gigantea needed for this study would be both inhumane and cost prohibitive (at least on my wallet).
I think company like DFS would have enough anemones availabe to do the type of study I have in mind. They do have a lot of anemones moving through their facility. Set up culture swabs for these are not likly to be expensive.

What kills cells on a petri dish may not work as a treatment in a QT setting, or may just kill the anemones. Aiptasia are more like gigantea than culture media.
We always use petri dish to test treatments of antibiotics. This method works very well for antibiotics. In complext animals like mamals, there are various bariers that keep antibiotics or other molecules out of certain area, and rapid degradation of these molecules (kidney, liver, avrious molecues that bind to and degrade medication). This is why there is differences between in vitro and in vivo outcome. I don't think we need to worry about this in anemones.

I am confused on this one. How do you think therapeutics are developed?

We need to ID what these infections are. How does model animals/anemones give us this information? We even know that other host anemones does not get infected or died as easy as Magnifica or Gigantea.
Can you think of anyway you can use model animals to give us this information?

[raythepilot]

I do not know about gigantea but I've treated BTA for bacterial problems with cipro.
I keep them in a small isolation tank and do 2 x 100% (morning /night) water changes per day. With each change I weigh in the correct amount of cipro. This treatment is 100% effective after 5 days. The problem is the anemone come out of treatment bleached because the cipro is toxic to the symbiotic algae in the nem. It takes about two weeks after treatment for the anemone to return to full color.

[OrionN]

Some of my Gigantea and Magnifica undergoes cipro treatment and did not bleached so I do not think the antibiotic, by itself is toxic to the zooxanthellae.

[raythepilot]

Do a google search on cipro as a plant killer.

[D-Nak]

I do not know about gigantea but I've treated BTA for bacterial problems with cipro.
I keep them in a small isolation tank and do 2 x 100% (morning /night) water changes per day. With each change I weigh in the correct amount of cipro. This treatment is 100% effective after 5 days. The problem is the anemone come out of treatment bleached because the cipro is toxic to the symbiotic algae in the nem. It takes about two weeks after treatment for the anemone to return to full color.

This is what I'm trying to determine. As Minh pointed out, some nems don't bleach. Could it be that the zoox is already dying or dead? On many occasions, treatment is accompanied by dead zoox but in a unique form where it appears binded ("rat poop"). This is not to say that the antibiotic is causing this binding.

[D-Nak]

Do a google search on cipro and a plant killer.

????

Can you provide a link?

[raythepilot]

????

Can you provide a link?

http://www.biology-online.org/articles/anti-anthrax-drug-kill-plants-too.html

[D-Nak]

http://www.biology-online.org/articles/anti-anthrax-drug-kill-plants-too.html

Thanks, this is very interesting.

[raythepilot]

This is what I'm trying to determine. As Minh pointed out, some nems don't bleach. Could it be that the zoox is already dying or dead? On many occasions, treatment is accompanied by dead zoox but in a unique form where it appears binded ("rat poop"). This is not to say that the antibiotic is causing this binding.

I'm not sure what you mean, but from my own experience using cipro, if the nem doesn't bleach the cipro is probably not effective. Cipro is a potent plant killer.

[OrionN]

I find it interesting also.
However, I like to pointout that zooxanthellae is more accurate described as endosymbiotic dinoflagellates. While they can be call an algale, they are not plants. They are not any closer to plan than the bluegreen algae which is more resemble a bacterial.
Zooxanthellaes are not even in the plante kingdom which all the plants belong to.

[raythepilot]

I find it interesting also.
However, I like to pointout that zooxanthellae is more accurate described as endosymbiotic dinoflagellates. While they can be call an algale, they are not plants. They are not any closer to plan than the bluegreen algae which is more resemble a bacterial.
Zooxanthellaes are not even in the plante kingdom which all the plants belong to.

You are right, I couldn't possibly know what I'm talking about after having done the experiment.

[OrionN]

You are right, I couldn't possibly know what I'm talking about after having done the experiment.
I am not sure if you been sarcastic or not.
I did sucessfully treated anemones with Cipro. Some bleach and other not. I don't know if Cipro kill plant or not, but this is beside the point as I pointed out that Zooxanthellae are not even plants. They are not even in the Plante kingdom.

I did find out that high dose prolong antibiotic are stressful to my Gigantea as I documented in my last Gigantea treated thread. The anemone did bleached. given everything, I cannot conclude that he bleached due to cipro.

[raythepilot]

I did find out that high dose prolong antibiotic are stressful to my Gigantea as I documented in my last Gigantea treated thread. The anemone did bleached. given everything, I cannot conclude that he bleached due to cipro.

How about this (My experiment):
Start with a bifurcated nem. Both pieces healed but showing signs of bacterial infection (ie inverting, giving off slime.)
Nem A is control (stays in tank). Nem B goes into holding tank. Water in holding tank is changed 2x per day and equivalent of 25 mg /gal cipro is weighed in each time. Treatment is for 5 days.
Piece A is not bleached and still has signs of infection. Piece B is cured and bleached.

[OrionN]

Ray,
Your experiment is full of holes. Do you really want me to poke at them?
The condition of the holding tank is obviously not the same as the DT. A better experiment would be taken them both to identical holding tanks (light, current, temp and water change regiment and same water source) the only different is the antibiotic.
Even with this you only have 1 trial. You cannot make sweeping conclusion with this finding.
You can also put 2 healthy anemone in optimal tank and treat them with Cipro on one and no Cipro treatment with the other. If you have multiple trials with this and cipro treated bleach every time, then you have something there.
As we found out DD treat their anemone with 7 days of Cipro at 250 mg/10 gal. Some of the anemone are bleach and other are healthy and not bleached. No control needed. Just this fact alone, would definatively show that bleaching is not due to Cipro.

[raythepilot]

Ray,
Your experiment is full of holes. Do you really want me to poke at them?
The condition of the holding tank is obviously not the same as the DT. A better experiment would be taken them both to identical holding tanks (light, current, temp and water change regiment and same water source) the only different is the antibiotic.
Even with this you only have 1 trial. You cannot make sweeping conclusion with this finding.
You can also put 2 healthy anemone in optimal tank and treat them with Cipro on one and no Cipro treatment with the other. If you have multiple trials with this and cipro treated bleach every time, then you have something there.
As we found out DD treat their anemone with 7 days of Cipro at 250 mg/10 gal. Some of the anemone are bleach and other are healthy and not bleached. No control needed. Just this fact alone, would definatively show that bleaching is not due to Cipro.

Of course, every nem that I treat this way gets bleached but I see clearly that I am wrong. Obviously you are using a more controlled method.
Are you sure that you have good cipro and a consistent level?
Could you try my technique and see what happens?
But then you don't have an unlimited supply of nem clones like I do.

[D-Nak]

Of course, every nem that I treat this way gets bleached but I see clearly that I am wrong. Obviously you are using a more controlled method.
Are you sure that you have good cipro and a consistent level?
Could you try my technique and see what happens?
But then you don't have an unlimited supply of nem clones like I do.

Banter aside, whether or not Cipro bleaches nems may be a non-issue as gigs that are treated with both Cipro and Septra are both bleaching (or not bleaching) but are still dying. I was wondering if the two were related, but I realize it will take a lot to prove or disprove the theory. We just don't know if what's bleaching nem is also killing the nem. Granted, we know that without a healthy zoox population, the nem will need to eat or will starve to death.

As I mentioned in my initial post, while it helps to hear about other species of nems, we really need to learn specifically about gigantea. BTAs readily clone, and they are a lot hardier than gigantea, so it's hard to compare the two species as to how each will react to Cipro or any other antibiotic. I've never had to treat ANY of my BTAs. I've treated almost all of my gigantea.

[raythepilot]

As I mentioned in my initial post, while it helps to hear about other species of nems, we really need to learn specifically about gigantea. BTAs readily clone, and they are a lot hardier than gigantea, so it's hard to compare the two species as to how each will react to Cipro or any other antibiotic. I've never had to treat ANY of my BTAs. I've treated almost all of my gigantea.

Your right! If we want to find something that cures a human disease do your study right in humans, Why mess around with mice. It's not the same.

Get my point.

If you have never had a sick BTA then you were pretty lucky. BTA suffer from the same decline disease as other species. In fact I've has healthy BTA's get sick from eating contaminated food. Had one for 6 months and went down in 2 months after taking frozen shrimp. If I had known about Cipro then I might have been able to save it.

Here is something that I think everyone here has a problem with. Crappy water. Yes bad water. My experience with BTA is that they are extremely sensitive to low levels of NH4+. You cannot test for the level of NH4+ that will kill a BTA.

I regularly do 2 x 25% water changes per week and often do 200% water changes per day when necessary.

You need that kind of water change to simulate a real reef condition.

[OrionN]

Of course, every nem that I treat this way gets bleached but I see clearly that I am wrong. Obviously you are using a more controlled method.
Are you sure that you have good cipro and a consistent level?
Could you try my technique and see what happens?
But then you don't have an unlimited supply of nem clones like I do.

It is easy for me to get Cipro for human rather for aquarium, but O do not think there is much different between the two.
I do think at the dose of 500 mg per 10 gal we reach near toxic level of Cipro for the anemone (Gigantea) I think 250 for 10 gal or 7.5 gal are more appropriate doses.

I think I will try to email Kevin with DD and see if he interested in trying to DI some of these infections.

[Amoo]

because the cipro is toxic to the symbiotic algae in the nem.

Since this is a thread for gathering facts, it should be pointed out that this is just not one of them. Here's an example of why, This .pdf shows the toxicity level of one of the drugs I have acquired to try, Cefotaxime.

http://www.phytotechlab.com/pdf/antibiotics.pdf

It also shows the toxicity levels to a lot of other commonly used drugs to treat bacterial infections.

What the article you linked states is that Cipro can plants. Okay, so can almost all of the drugs listed on the table, but guess what, they are all used in some form of function via testing/treatment.

I have treated both a GBTA and a Purple Mag with 250mg/10G of Cipro for 7 days and have not experienced bleaching in either. Many others have treated and not experienced bleaching. Trying to state Cipro is toxic to Zoox and linking an article that says Cipro can be toxic to plants is just not going to cut it in regards to the types of info we're going to need.

Here's a study where Vanco, Carbenicillin and Cefo were used on multiple plants, again all three are toxic beyond certain levels:

http://www.horticultureresearch.net/pdf/Silva.pdf

To assume Cipro is any different is nothing more then an assumption of which the article you linked does nothing to dissuade.

Here's a Cipro study actually related to aquatic plants:

http://www.ncbi.nlm.nih.gov/pubmed/21919043

Or you can just get to the crux of the matter which is discussed here:

Fluoroquinolones
(FQs) like ciprofloxacin, lomefloxacin, ofloxacin, and
enrofloxacin are a class of antibiotics. They are commonly
used in both human and veterinary medicine, and are not
readily biodegradable by microorganisms [1]. At low con-
centrations, FQs are phytotoxic and toxic to most plants and
animals. At the same time, while water plants respond to
FQs with chlorosis, due to chlorophyll synthesis inhibition
fish are not affected even by high concentrations [2].

http://www.pjoes.com/pdf/22.1/Pol.J.Environ.Stud.Vol.22.No.1.71-76.pdf

The above was taken from an article published 3 years after the one you linked.

So we have learned that FQs, specifically Cipro does have some toxicity level to plants, as most drugs do. It also will inhibit chlorophyll synthesis. Nowhere have we learned that Cipro kills Zoos below or above a certain level.

Keep in mind also, that we are currently treating 250mg/10g regardless of size OR type of specimen. We don't have any specific guidelines in place based on size or kind of nem, it's just a sweeping "get the water to this concentration".

[DasCamel]

Your right! If we want to find something that cures a human disease do your study right in humans, Why mess around with mice. It's not the same.

Get my point.

If you have never had a sick BTA then you were pretty lucky. BTA suffer from the same decline disease as other species. In fact I've has healthy BTA's get sick from eating contaminated food. Had one for 6 months and went down in 2 months after taking frozen shrimp. If I had known about Cipro then I might have been able to save it.

Here is something that I think everyone here has a problem with. Crappy water. Yes bad water. My experience with BTA is that they are extremely sensitive to low levels of NH4+. You cannot test for the level of NH4+ that will kill a BTA.

I regularly do 2 x 25% water changes per week and often do 200% water changes per day when necessary.

You need that kind of water change to simulate a real reef condition.

An ammonia badge starts to detect at .02, I had a little spike adding fish back into my system and noticed it. Ever since then my bubble tip has been on a slow death spiral. My others Nems were irritated and bounced back rather quickly. Some Amquel resolved the issue as the tank naturally balanced out. The ammonia badge was very key. I agree they really don't like ammonia.

[taylor t]

I'm not familiar with all the medical lingo. But, what I do know, is I've treated gigs with a bottle marked as "aquatic cipro" (that makes PEOPLE sick, don't ask, it's different than pharmacy grade, I found out first hand) that has stopped the downward spiral of the gigs I've had to treat, and not all of them have bleached, and then have them recovered. To me, it doesn't matter what one calls it. I've watched them recover and not lose color with treatment, more than once. Have they lost color? Some, but not every time. If it was truly meds, it would happen every time. My test pool (me and Pete combined is over a dozen), is only a small batch of tested gigs with meds. For us, the meds work, and don't bleach every time, maybe it's a placebo? Not sure, but makes humans feel sick that's for sure. Either way, for us, it was the missing link we didn't have prior. I've also always been an advocate of 100% water changes every day when trying to get them to take, and I've found, how I make my water matters, for me anyways. Pulling off the 50 gallon container sitting doesn't have good results for me, I needed to make fresh every 24 hours with a power head in a 5 gallon bucket. Consistent. Gigs get to use water from hours 24-48 after making. Then change. Just how I did it with good results. Maybe, just the water changes makes the difference, and the meds don't matter. I don't know. What I think is they have problems purging from the shipping, on top of lacking flow and light for so long. Two other factors that play a big part. No matter how you see it, gigs are a TOUGH anemone to get to stabilize from import. A month won't do it, it can take that long just to show signs of dying. But to me, it seems once they are well "acclimated", they are pretty durable, more than sps corals anyways. Still sensitive don't get me wrong, but not hyper sensitive like in the beginning. I would never consider BTA as hyper sensitive, even when cut.

A few of our gigs came in already cut, as freshly cutting them would definitely add stress, which could be a factor too. I've cut BTA's also, and had one half act one way, and the other half act differently, same water. More than once. There is night and day difference between gigs and BTA, so how BTA's react isn't even a factor in my book. I've treated gigs together and had them both act differently. That's what makes our efforts in gig recovering so difficult, it's not a "one size fits all", we could talk all day about it and still walk away baffled. I currently have 2 purple gigs, both came in cut, both are recovering differently as one is nice and dark with the mouth becoming more centered (almost looks like it wasn't cut), the other is not as dark, still looks like it's partially bleached, and the mouth is still on the edge. Both treated the same, and both "stable", but far from acclimated, 5 months later.

I would consider only 2 (or 3) of my 7 gigs "well acclimated" ( I would consider it 3, but one of them was a cut in the beginning, and I'm not sure it would survive a cut again). The rest are just "stable". IMO, cutting a "stable" gig is a dead gig. Cutting an "acclimated" gig has a chance, IMO, but not a chance I'm willing to take, yet. Honestly, I bought one of the last 4 I added for a trial cut, but it takes so long for them to acclimate, not just become "stable". It takes sooooooooooooo loooooooooong for them to acclimate, yet, they can go downhill so fast. Just my .02.

[Fishmommy]

I would like to beg of you who treat with antibiotics - please dump your water change water and don't reintroduce it to the water supply.

[OrionN]

.......
Keep in mind also, that we are currently treating 250mg/10g regardless of size OR type of specimen. We don't have any specific guidelines in place based on size or kind of nem, it's just a sweeping "get the water to this concentration".
The total fluid volume of an average size human is about 10 gal. This is very rough calculation on my part. Assuming that the medication we use have a complete distribution to all water volume in a human, given a single dose for human use in 10 gal should get the concentration to where the medication is effective to treat the infection.
Factors that increase or decreased concentration in certain area due to barrier we have in our body (not all the medication are absorbed, Medication does not get into some of the cavities well like joints or to the brain because of blood brain barrier, medication are concentrated to the kidney and bladder because it is actively eliminate into urine....)

In land animal the volume of distribution of the medication is limited to the water volume of the animal. That is the reason why dose of medication depends on the size of the animal. In the fish tank, on the other hand, the volume of of distribution of the medication is the tank volume therefore the recommended does depends on the water volume of the treatment tank.

I have tried the medication dose for my anemone first to see if there is any clear cut problem before I recommended it. In case that I guess and not used that dose before personally, I try to mention that this is the case.


I would like to beg of you who treat with antibiotics - please dump your water change water and don't reintroduce it to the water supply.
In both ciprofloxacin and trime/sulfa, in human use, our body excreted the medication unchanged into the urine thus into the sewage system. Both medication are photo degraded and get broken down by sunlight. I assume that in animal use, they eliminated thought the urine into the environment, and there it get degraded by the Sun.

[Amoo]

Minh,

I understand where you get your volume calculation from and honestly have no issues with it. The one area though where I feel it may be revised eventually (or may not) is dosage via species. As we can see in some of the studies I link and others that are out there a plant is not a plant is not a plant. Many show different tolerances to some of these drugs and I'm wondering if Nem species are the same.

[OrionN]

I am hoping that the needed concentration is not anywhere near the toxic concentration. The information we need is what species and the sensitivity of the pathogens.

[taylor t]

(I'll attempt humor once more, last time it came out as disaster, I'm sorry...)
(humor here)
I'm not the sharpest knife in the drawer. I'm, ok with that. I'd rather have fun. Clearly, all the health care professionals understand it much more clearer, all I'm reading (humor here, remember :) )is "bla bla bla gram negative bla bla gig". But, is it possible, (humor) that once you find out the "patient" has hepatitis "c", even giving it the proper medication and the proper grams, still doesn't address the issue that they walked into the ER with a plastic bag stuck down it's throat? or has bad sun burn with pealing skin that needs something more than a band aid? Or ate too much the night before and went straight to bed? (all humor here)




(ok, end of humor)
I don't know a lot of things. But just looking back on my own share of gigs I've killed, I've realized a lot of things I wish I did differently now. I think there's a LOT of other things that kill them as well, and I think meds are looked at as the "skapegoat" as mentioned before. Even DSF kills their share, and they're doctors! No disrespect to anyone, as it's a requirement to start with a good one first... We all have understood this for a while now. No one can save one that's already internally decided to die. Finding out if it's positive or negative or neutral, may help a few, but most likely not the majority, still. I still think there is some luck in getting one that hasn't "checked out" already, which could take a month or longer sometimes to do so, no matter how many water changes are done. D-Nak's an accomplished reefer, and he was able to save 1 of 4. He has YEARS of gig experience too. Luck of the draw sometimes. Gigs don't travel well.

I look at the difference of flow with them, and, let's say a BTA. You blast a BTA with flow(I've done it), and they will move away, usually. I blast a gig, it stretches TOWARDS flow, again, usually. Always an exception. Is it possible, that it's not an infection, but a structural difference inside we just don't know or understand? Maybe gigs can't move water internally on their own, fast enough, (which requires high flow to move it around for them), and maybe a BTA can, or doesn't need as much as fast because it's "metabolism" is slower?. Maybe some slow current early upon receiving a gig, so the gig can "understand (if you can appease me a moment) when they first get there that it's ok to inflate, then slowly ramp it up to what it really needs long term? (HUMOR) before they start getting light headed (maybe I'm the one that's light headed, I don't know). Maybe it's in dire need of some blasting to get "the ol' pipes moving again". Maybe dead zoox clogs it's internal "pipes" upstream internally at times. Just a thought, we've all been constipated if you're honest.

Then there's light. Could be the patient gets sunburned really easy, on day one, out of the truck container. Who knows what it was getting before? Was it under a ledge, up against some tall grass only getting 5 hours of direct light, or on a rock getting 14 hours??? Blast too much early, sunburn and shrink. Not enough, it will kill the zoox it already has, if it's not in the process of clearing the dead zoox already... Uh oh, here comes constipation again... Then fast ramp up of intensity, then sun burn again... "Why is it shrinking???" ..."I don't know?".... Sounds like a Brian Regan comedy show. (We have lots of laughs in our family, if you haven't grasped that yet! I will try to contain myself from here on out).

Then there's water. Could be it's too drastic of an elevation change, (PH) like going from Florida sea level, to Colorado overnight, or to a smog air filled city, and then asked to go jogging like the day before. We'd all deflate too.

The above are just some silly (and some real) thoughts. No disrespect to anyone.. :) Maybe something I've said will spark some thought in the guys that understand biology better than me, I'm not schooled (I earn money turning screwdrivers). Even figuring out what virus they get, who's to say one comes with bird flu, one with common cold, and the next shipment comes with cancer from the beach they were living on that dumps toxic chemicals in the water a mile away. TOOOOOO many variables with all of this. I think it would be more productive to show what a thriving gig looks like, what they all seem to like when they are thriving, and for those with gigs to explain their own journey how their particular gig made it "over the hump", and struggles they have had to overcome, I know I've had mine. EVERYONE has issues at times. Even Minh has had to treat his purple after being in captivity for so long, and he's the treatment pioneer! I may need to treat one of mine again in the future too. My green 210 gig was treated 4 months ago, for probably the tenth (who knows) time in captivity, I'm the third reefer to treat it. I've found they all are a little different initially. Bottom line, gigs are a TOUGH anemone to acclimate... Gosh, I've said too much, I can't believe I'm posting this.... Grace please. Best of luck to everyone. :)

[D-Nak]

I should've been more specific when I introduced the thread. The assumption here is that the gigs we're dealing with are newly imported and sick, from what we don't know for sure, but going on the assumption that the gig has a bacterial infection. In other words, we don't mean to use the infection as a scapegoat for not properly acclimating an otherwise healthy gig, but that we need to figure out a new way to treat gigs that are obviously sick. Yes, there are a lot of variables to contend with, and not only is it going to take a lot of time, but we'll also some luck to figure this out.

I completely agree with you when it comes to flow, lighting, and water. These are variables that we need to study. While we frequently have gigs in our tanks literally blasted with water, most of the photos I see of gigs in the wild are in lagoons with minimal flow. Many times, these same photos show the gigs exposed to the sun during low tide. This causes us to assume that gigs can be blasted with light. However, during the collection, export, import, and final sale, we don't know what conditions the gigs are in. To cause bad bleaching like we've seen, we're assuming that the gigs are without light or are in low light environments for a long time. Regarding water quality, again referring to the photos of gigs in the wild, many are in turbid water, which we can assume is nutrient rich.

But, these are photos of healthy gigs. So, we need to determine what constitutes optimal conditions for a SICK gig. Water quality is easy -- we just start with clean water. Flow is a bit more difficult to determine, as you pointed out. I completely agree with you that sick gigs need enough flow to help them purge the dead zoox ("rat poop") but not too much flow where they are using energy to fight a current to remain upright or attached. Constipation is the same word I would use, and have used, to describe that it looks like the gig is experiencing. Healthy gigs don't expel rat poop. Regarding lighting -- this is a tough one. Not enough light, and the remaining zoox could die. Too much light and the gig could react negatively. For example, IME many gigs deflate during the later part of the day, after they've received an ample amount of light. This leads me to think that the zoox population's fuel tank is full so to speak, and to stop the process, the gig deflates. However, this doesn't happen 100% of the time. My last few gigs were deflated in the morning, then inflated with lights on.

So, in short, while getting to the root of the issue in terms of the sickness itself is important, I agree that we also need to study environmental conditions. Just another piece of the puzzle. And yes... too many variables!

(I'll attempt humor once more, last time it came out as disaster, I'm sorry...)
(humor here)
I'm not the sharpest knife in the drawer. I'm, ok with that. I'd rather have fun. Clearly, all the health care professionals understand it much more clearer, all I'm reading (humor here, remember :) )is "bla bla bla gram negative bla bla gig". But, is it possible, (humor) that once you find out the "patient" has hepatitis "c", even giving it the proper medication and the proper grams, still doesn't address the issue that they walked into the ER with a plastic bag stuck down it's throat? or has bad sun burn with pealing skin that needs something more than a band aid? Or ate too much the night before and went straight to bed? (all humor here)




(ok, end of humor)
I don't know a lot of things. But just looking back on my own share of gigs I've killed, I've realized a lot of things I wish I did differently now. I think there's a LOT of other things that kill them as well, and I think meds are looked at as the "skapegoat" as mentioned before. Even DSF kills their share, and they're doctors! No disrespect to anyone, as it's a requirement to start with a good one first... We all have understood this for a while now. No one can save one that's already internally decided to die. Finding out if it's positive or negative or neutral, may help a few, but most likely not the majority, still. I still think there is some luck in getting one that hasn't "checked out" already, which could take a month or longer sometimes to do so, no matter how many water changes are done. D-Nak's an accomplished reefer, and he was able to save 1 of 4. He has YEARS of gig experience too. Luck of the draw sometimes. Gigs don't travel well.

I look at the difference of flow with them, and, let's say a BTA. You blast a BTA with flow(I've done it), and they will move away, usually. I blast a gig, it stretches TOWARDS flow, again, usually. Always an exception. Is it possible, that it's not an infection, but a structural difference inside we just don't know or understand? Maybe gigs can't move water internally on their own, fast enough, (which requires high flow to move it around for them), and maybe a BTA can, or doesn't need as much as fast because it's "metabolism" is slower?. Maybe some slow current early upon receiving a gig, so the gig can "understand (if you can appease me a moment) when they first get there that it's ok to inflate, then slowly ramp it up to what it really needs long term? (HUMOR) before they start getting light headed (maybe I'm the one that's light headed, I don't know). Maybe it's in dire need of some blasting to get "the ol' pipes moving again". Maybe dead zoox clogs it's internal "pipes" upstream internally at times. Just a thought, we've all been constipated if you're honest.

Then there's light. Could be the patient gets sunburned really easy, on day one, out of the truck container. Who knows what it was getting before? Was it under a ledge, up against some tall grass only getting 5 hours of direct light, or on a rock getting 14 hours??? Blast too much early, sunburn and shrink. Not enough, it will kill the zoox it already has, if it's not in the process of clearing the dead zoox already... Uh oh, here comes constipation again... Then fast ramp up of intensity, then sun burn again... "Why is it shrinking???" ..."I don't know?".... Sounds like a Brian Regan comedy show. (We have lots of laughs in our family, if you haven't grasped that yet! I will try to contain myself from here on out).

Then there's water. Could be it's too drastic of an elevation change, (PH) like going from Florida sea level, to Colorado overnight, or to a smog air filled city, and then asked to go jogging like the day before. We'd all deflate too.

The above are just some silly (and some real) thoughts. No disrespect to anyone.. :) Maybe something I've said will spark some thought in the guys that understand biology better than me, I'm not schooled (I earn money turning screwdrivers). Even figuring out what virus they get, who's to say one comes with bird flu, one with common cold, and the next shipment comes with cancer from the beach they were living on that dumps toxic chemicals in the water a mile away. TOOOOOO many variables with all of this. I think it would be more productive to show what a thriving gig looks like, what they all seem to like when they are thriving, and for those with gigs to explain their own journey how their particular gig made it "over the hump", and struggles they have had to overcome, I know I've had mine. EVERYONE has issues at times. Even Minh has had to treat his purple after being in captivity for so long, and he's the treatment pioneer! I may need to treat one of mine again in the future too. My green 210 gig was treated 4 months ago, for probably the tenth (who knows) time in captivity, I'm the third reefer to treat it. I've found they all are a little different initially. Bottom line, gigs are a TOUGH anemone to acclimate... Gosh, I've said too much, I can't believe I'm posting this.... Grace please. Best of luck to everyone. :)

[taylor t]

Ok. Suppose a gig has everything it needs to thrive. Acceptable water, flow, temp, and light. It continues to go downhill in the exact same water quality a healthy gig is stabilized in. Someone who understands biology may be able to answer this. If a medical person was to take a swab from the deteriorating anemone, by swabbing the gaping mouth, find out what the bacteria was living inside, advise on the appropriate antibiotic, I'm wondering. IF, that deteriorating anemone DID NOT have an infection of sorts, (and had internal structural issues in it's handling, or respiratory issues from lack of "x,y, or z" for some time), and antibiotic "x" was dosed because the bacteria inside was needed for the anemone to live. It would kill the anemone. Right? Maybe a swab of a good, healthy anemone should be done to see what is living at "ground zero", so there's some sort of baseline that you DON'T want to kill. Then, find a deteriorating anemone to swab, and compare, no?

And, just throwing this out there, maybe way out I don't know... It could also be possible that there is a structural "issue" that causes gigs to degrade from lack of something, whether it's flow or light, or build up of x,y,or z for too many hours. There could be just a natural process of accelerated decomposition internally, because of the nature of salt water, no? When we see them in nature, they are in calm pools, yes. But, it's a constant taking away of "waste water" away from the anemone, where they don't get that benefit in our little glass boxes. Maybe that's why they do so well with high flow in our tanks, they can't handle a waste product they are excreting. When I kill flow to my tanks, they puff up and look beautiful, for a time. Too long with no flow, they go down, whether it's an hour or a day(s) (in our tanks I'm talking). About 2 weeks ago, my green 210 gig went flat, for a couple days. I almost pulled it, but decided to "hammer" it with flow. It had gaping mouth and flappy. It was getting good flow on most of it, but had one side that was tucked up in the rocks with almost no flow for the last couple months, tents were short on the back of the nem, shorter than the flowing front area. That's what got me thinking, maybe there's a structural issue they have with not being able to "pump" water around inside themselves, like other anemone's. (In the animal kingdom, some species have differences between groups, right?) So I left it, added 2 more power heads next to it, raised it up so the foot got blasted and the part that didn't get flow was flapping in the flow, and with in a couple hours, it was back to where it was. I was shocked how fast it inflated, after being floppy so long. I really thought it was going to need meds agian, all the baby clowns were rubbing it for a day or two while it was flat. After flow was added, it went back to normal. Not sure of the relevance of all this, just throwing it out there. Maybe we're looking for a structural difference, or a bacterial difference, or both. BUT, IF, there IS a bacterial issue, wouldn't we need a "ground zero" starting point we DON'T want to upset? We may be looking at 2 issues, and that's why we struggle, we're looking at the problem as 1 issue. Just a thought.

[taylor t]

And to add, if gigs can't move water by themselves, because the internal "pump" isn't working, could that add to the problem that some don't respond to antibiotics? Because they aren't getting the drugs inside them to do the job?

[raythepilot]

Here's a Cipro study actually related to aquatic plants:

[url]http://www.ncbi.nlm.nih.gov/pubmed/21919043


This is interesting!
In this study, the EC50s for D. subspicatus was 8,042 µg/L for ciprofloxacin which corresponds to about 30 mg/gal.
This is very close to the recommended dosage of 25 mg/gal. which may account for the variable results found by some people.

Based on this report, cipro should not be used above 3 mg/gal unless you can tolerate some algae death.

This confirms my opinion that if you do not have bleaching, you probably do not have an effective dose of Cipro.

[Amoo]

This is interesting!
In this study, the EC50s for D. subspicatus was 8,042 µg/L for ciprofloxacin which corresponds to about 30 mg/gal.
This is very close to the recommended dosage of 25 mg/gal. which may account for the variable results found by some people.

Based on this report, cipro should not be used above 3 mg/gal unless you can tolerate some algae death.

This confirms my opinion that if you do not have bleaching, you probably do not have an effective dose of Cipro.

...facepalm

[taylor t]

Now, for the slow kid in class, does this mean, most of my gigs have not been treated properly, and nothing more than a fresh saltwater purging is what made them better??? That most of what we've treated for, was just saltwater changes with a dab of selective strain strengthening? (because nothing really died?)

This would mean, we're right back at square one, with the exception, we've learned they have difficulty purging all the old water inside when we first get them.

[DasCamel]

And to add, if gigs can't move water by themselves, because the internal "pump" isn't working, could that add to the problem that some don't respond to antibiotics? Because they aren't getting the drugs inside them to do the job?

I give all my sick Nems massages until they inflate and can expel/move fluids on their own. Circulation of fluid is definitely a factor. Using a turkey baster to blast them a bit also works.

[taylor t]

raythepilot, are you saying, you have better survival rates with BTA's by using cipro, and bleaching them, than not? Now I'm wondering, if much of the "success" we've seen, or unsuccess we've seen, has been determined by the amount of, and process of water changing alone, or lack there of. With exception to the anemone's that have bleached, only making their recovery more drawn out than it needs to be? Then I further wonder, if establishments that are using Cipro, are actually creating more problems, ADDING to the equation drug resistant pathogens. Thoughts?

[raythepilot]

I am not sure about mags or any other species but here is what I know from long years of experience with BTA.
Water is the first thing. If any of my nems look sick I do lots of water changes. I am very careful about how I make my water. It has to be 35.0 ppt +/_ 0.1 ppt before I use it and it has to be within +/-0.2C of the ambient tank temp before I add it to the nem tank.
I regularly do 2 x 50% water changes per week and more if anything doesn't look right.
This is my baseline and when ever I check parameters they are dead on.

Recently when bifurcating the animals, some got infected and displayed the typical symptoms of nem decline. I've successfully treated these with cipro at 25 mg./gal. but they bleach.

I reseeded these anemone with flora from some healthy anemone and they recovered in about 2 weeks. One of these was in the pair that recombined in my post on the subject.

[taylor t]

Thanks raythepilot. I appreciate your insight.

[Amoo]

Now, for the slow kid in class, does this mean, most of my gigs have not been treated properly, and nothing more than a fresh saltwater purging is what made them better??? That most of what we've treated for, was just saltwater changes with a dab of selective strain strengthening? (because nothing really died?)

This would mean, we're right back at square one, with the exception, we've learned they have difficulty purging all the old water inside when we first get them.

Not necessarily, honestly we just don't know. The part he quoted was for a green FW alagae. That same article talks about the toxicity level of Cyano to the same drugs and they are more sensitive.

All we know is that Cipro has a photosynthesis inhibition component. Every plant/algae...etc that has been tested shows different results in regards to toxicity. We know that the nem itself is a "marine plant" and they have a symbiotic relationship with their Zoox. Ironically we believe but don't know that different species of Nems don't infect each other, but they MAY have chemical warfare.

All of these things would lead me to ask if the Zoox inside a BTA is the same as a Zoox inside a Gig as inside a Mag...etc. The answer here is we don't know. It is entirely possible that the Zoox inside a BTA has a different toxicity level then the zoox in a Gig. BTAs may be more susceptible to FQs then Gigs...etc. I believe raythepilot has bleached every BTA he has treated. I also believe I didn't bleach the 8" one I treated (or the Mag). I use the same dosage he does and I have a medical grade Cipro so I know the dosage is spot on. Was there a different in our flow, temperature, lighting that could have an effect on the bleaching? My guess would be yes. Could those factors combined with a photosynthesis inhibitor cause his to bleach and mine not to? Absolutely.


The important thing here is we are treating the bacterial infection and trying our best to do as little harm as possible to the Nem, kind of like Chemo in a way. The important thing is we administer a high enough dose to kill the bacteria while doing as little harm as possible to the Nem (none ideally, but we can see from bleaching that isn't always the case).

PilotRay seems to treat a lot, he is in a better situation then most of us to experiment with lowering his dosage to see if he can still kill the bacteria and not bleach the nem. Move down to 20 or 15 or 10...etc. Again though I firmly believe 5-10 years from now when we have a more clear picture of what's going on, we're going to find we have different dosages for different species.

[clorox]

We need to set up a Kickstater campaign to sponsor the study and rehabilitation of anemones ;)

[OrionN]

I have been reading some papers on culture bacteria from sea anemones. The problem is that the normally acceptable method of culture result in death of the anemone.
In one study, the anemone was washed in steril salt water then a 1cm2 of the endodem was aseptically removed, pulverized. The result liquid then was cultured.

Anyone know of any method of culture bacterial from anemone? I am thinking of deep mouth swab or aspirated with a steril syrine.

[clorox]

I have been reading some papers on culture bacteria from sea anemones. The problem is that the normally acceptable method of culture result in death of the anemone.
In one study, the anemone was washed in steril salt water then a 1cm2 of the endodem was aseptically removed, pulverized. The result liquid then was cultured.

Anyone know of any method of culture bacterial from anemone? I am thinking of deep mouth swab or aspirated with a steril syrine.

Did you look at this?

http://www.ncbi.nlm.nih.gov/pubmed/18405113

It doesn't describe the methodology, but from it you might be able to trace papers that might.

[TinManx]

I have been reading some papers on culture bacteria from sea anemones. The problem is that the normally acceptable method of culture result in death of the anemone.
In one study, the anemone was washed in steril salt water then a 1cm2 of the endodem was aseptically removed, pulverized. The result liquid then was cultured.

Anyone know of any method of culture bacterial from anemone? I am thinking of deep mouth swab or aspirated with a steril syrine.

A model system would come in handy .... lol

[OrionN]

A model system would come in handy .... lol
How can a model system help tel us what bacteria infect Gigantea or Magnifica?

[TinManx]

How can a model system help tel us what bacteria infect Gigantea or Magnifica?

I would personally rather put aiptasia in a blender than gigantea. Much of the work in this area requires destruction, or potential destruction, of the animal of interest. In situations like these, most people who do this for a living will opt to work with a model organism. Why do you think coral scientists transfected vibrio sp. to aiptasia in the first place? Aiptasia are free, they reproduce rapdily, are clones (same genes), and have been genitically characterized.

[OrionN]

Still I don't see how you can use model for these information.

We use models when we know the cause of the disease which does not have to be infectious. We find a model with this disease, or cause them to get the disease and try treatment protocols.
In human, we find a model like Rhesus Monkey, or Chimpanzee, give them the disease and try treatment protocols on them. Obviously we should not give another human the disease then try treat (this did not stop some people in the past like in the case of Japanese researchers during WWII). Another way we use model is to give it to the animal and follow and study the natural history of the disease. Some US researchers did this to human in the past too regarding Syphilis infection. Although the researchers did not give a group of prisoners Syphilis but they diagnosed prisoners with this disease. The researchers then follow and did not treat the prisoners so they can observe the natural history of this disease. Because of the ethics involved, models are only useful in human diseases. Chimpanzee DNA is about 98% the same as human DNA. Aptasia is not anywhere similar enough to Magnifica or Gigantea to even remote consider as a model for these two more valuable anemones.

I only briefly scan the article you mentioned. Notice that they injected Vibrio into the Aptasia and notice that they get infected and died. I thought the paper was pretty useless, at least for me. I have no idea why they do this. I cannot think of any application that this can be use or or any useful information can derive from this experiment. The result was so obvious that if the animal involve is anything else other than the lowly Aptasia, it would be unethical waste of life IMO.

[raythepilot]

Did you look at this?

http://www.ncbi.nlm.nih.gov/pubmed/18405113

It doesn't describe the methodology, but from it you might be able to trace papers that might.

I took me a while to get the complete paper because it is crossed reference in the data base at the J. Environ. Bio.

But the paper is not too encouraging. According to the authors, the studied anemone Stichodactyla haddoni has a number of normal flora bacteria that provide the host with resistance to human and fish born pathogens. They are like the flora found in our guts.
I guess when you treat nems with a broad spectrum antibiotic like Cipro, you kill off their gut bacteria and leave them in even worse shape than before.
I guess when I inoculated my bleached nems with some tissue from a healthy nem I was also reintroducing the normal flora they needed.

It makes sense actually.

I guess what you need are some healthy gigs. Then when you kill off everything in a new gig with cipro you can add back the healthy flora.

But they can still be infected with bacteria from sea source food.

[TinManx]

Still I don't see how you can use model for these information.

We use models when we know the cause of the disease which does not have to be infectious. We find a model with this disease, or cause them to get the disease and try treatment protocols.
In human, we find a model like Rhesus Monkey, or Chimpanzee, give them the disease and try treatment protocols on them. Obviously we should not give another human the disease then try treat (this did not stop some people in the past like in the case of Japanese researchers during WWII). Another way we use model is to give it to the animal and follow and study the natural history of the disease. Some US researchers did this to human in the past too regarding Syphilis infection. Although the researchers did not give a group of prisoners Syphilis but they diagnosed prisoners with this disease. The researchers then follow and did not treat the prisoners so they can observe the natural history of this disease. Because of the ethics involved, models are only useful in human diseases. Chimpanzee DNA is about 98% the same as human DNA. Aptasia is not anywhere similar enough to Magnifica or Gigantea to even remote consider as a model for these two more valuable anemones.

I only briefly scan the article you mentioned. Notice that they injected Vibrio into the Aptasia and notice that they get infected and died. I thought the paper was pretty useless, at least for me. I have no idea why they do this. I cannot think of any application that this can be use or or any useful information can derive from this experiment. The result was so obvious that if the animal involve is anything else other than the lowly Aptasia, it would be unethical waste of life IMO.


Thank you for that! Very educational!

[OrionN]

Did you look at this?

http://www.ncbi.nlm.nih.gov/pubmed/18405113

It doesn't describe the methodology, but from it you might be able to trace papers that might.
I read some of the papers. The normal methods of culture bacterial from anemones is to wash the anemone several times in sterile sea water. The anemone is the cut up, pulverized and the liquefied anemone juice is then plated out, or culture in broth. Some study separated the area/organs of interested (tentacles, body, endometrium...) then pulverized it and culture. In all the studies I read, culture required scarified the anemone.
I also read online notebook of some students somewhere for a upper division biology course (Labs or field study) They collected the anemone, wash them then irritated it by wash in sterile pure water, collect the mucus and culture. Swab the outer surface and culture..... I am not sure if this can be use.

Anybody who find any method of obtain culture from anemones please let me know. Post it here or send me a PM or email would be great. I don't have full access to some of the scientific papers online, so sometime it is difficult for me to get full text of some of these papers. Sometime I just have to pay for the paper.

[OrionN]

I can set cultures and test for sensitivity but don't have exposure to newly import anemones. If I can get somebody with access to import anemones to obtain samples, I can culture them for the pathogens.
Brain storming on how to culture anemones, I came up with these three methods.

1. Get a syringe with large bore needle and stick blindly into the anemone and aspirated the juice for culture. This certainly will stress and may cause significant damage to the anemone. I know that for healthy Tridacna clams, biopsy like this does not cause significant problem. I think for healthy anemone, this should not cause significant problem. I am sure that this does cause significant stress to the anemone and may kill a sicken anemone.

2. Do a deep swab inside the mouth of the anemones. This is not likely to cause significant stress but there may be contamination and we may grow something that is not the actual pathogen.

3. Take the anemone out, wash him with sterile salt water, then and put him in a minimal amount of sterile sea water. Get him to contracted, then culture the solution we get.

I think the most promising method will be aspiration of the fluid inside the anemone, but I am too chicken to try it on my anemones at this time. What do you guys think? Any other idea? If I can come up with a, not too invasive, method to culture the anemones I will try this on my anemones to see what I get.

[raythepilot]

I can set cultures and test for sensitivity but don't have exposure to newly import anemones. If I can get somebody with access to import anemones to obtain samples, I can culture them for the pathogens.
Brain storming on how to culture anemones, I came up with these three methods.


Since you are dead set on this, why don't you try this:

According to the Ebert paper, they found that the level of bacteria was higher in the tentacles than the body tissue. They found about 2.5 x 10^4 CFU per gram in the tentacles vs 2.0 x 10^4 CFU per gram in the body.

If you send someone a few tubes of sterile saline, they could clip off a few tentacles from the nem and add it to the tubes and send them back packed in ice by Fed Ex. I used to do that all the time and it always worked. We even set reference cultures around like that.

[raythepilot]

Here are the normal flora you may find:

The associated bacterial
counts were maximum in tentacle tissues than in body tissue (Fig.1),
which were identified as Alcaligenes sp, Corynebacterium sp,
Aeromonas sp, Sporosarcina sp, Renibacterium sp,
Carnobacterium sp1, Carnobacterium sp2 and Salinococcus sp

[OrionN]

I don't want the normal flora. I wan to culture the pathogen, if any. I will culture healthy anemone also so that when we recover an organism, we have a idea if this is the pathogen or not.

[raythepilot]

I don't want the normal flora. I wan to culture the pathogen, if any. I will culture healthy anemone also so that when we recover an organism, we have a idea if this is the pathogen or not.

How will you know if it is a pathogen? Infect another nem?

[OrionN]

How will you know if it is a pathogen? Infect another nem?
It is a guessing game aid by statistic.

Speaking hypothetically, if I get samples from 10 healthy anemones and culture them. If I recovered 5 species of bacterial (A, B, C, D, E) in these 10 anemones, and all five are in about 80% of the anemones. This is a clear cut case of normal flora.
Also if we quantity and do a colony count per ml, I would expect very sick anemone will have a much higher pathogen bacterial count per ml (say 50,000+ per ml), then a normal flora count (<20,000 per ml). These number just got throw out from my head as examples. I just have to collect a bunch of cultures, detail/pictures of the anemones when and from which the culture from. Details before, any treatment pre/post culture and out come. From these information, than I just have to try to draw accurate, meaningful conclusions from the data.

Knowing the information above, then if I culture a sick anemone and come up with A, B, C, E, F and colony count of all strain except F is 10,000 per ml. Colony count of F is 100,000 colonies per ml. This is a clear cut information that show F is a pathogen and A, B, C, and E are normal flora.

Analyze and come up with meaningful and accurate conclusions is a huge part of an experiment.

[raythepilot]

It is a guessing game aid by statistic.



Sounds like a real research project! Do you get another degree when you publish. :beer:
You probably know this but you can homogenize the nem parts with a tissue grinder and saline then plate 0.1ml directly on media.

Good luck/skill.

[clorox]

I think the most promising method will be aspiration of the fluid inside the anemone, but I am too chicken to try it on my anemones at this time. What do you guys think? Any other idea? If I can come up with a, not too invasive, method to culture the anemones I will try this on my anemones to see what I get.

You know who will do it for you.....hint...hint :)

[Amoo]

Obviously the least intrusive method here would be the fine-needle aspirate. Unfortunately I'm not sure it would be the most effective in what we are trying to accomplish. It could, but you would need to develop very exact protocols which may be impossible to follow. For example:

You would always want to ensure you aspirate from almost the exact same location on every specimen, withdrawing the exact same amount of fluid. This I feel is going to be very tough going from species to species or even specimen to specimen. Again, I think it could be effective, but it may be damn hard to draw say 5ml of fluid out of deflated nem assuming you want the fluid from inside. I also think it would be too difficult to get people to withdraw the proper amount of physical sample while posing a much greater risk to the Nem itself (Stabbing into the stomach...etc.)

The next best and possible ideal option to accomplish what "we" are trying to do here, would be to cut off "x" inches from one tentacle. This may sound cruel, but how many of us already do something similar to repopulate Zoo in a bleached nem. Developing a protocol to have tentacle samples of "x" length sent to you seems like the way to gather the most information with the least harm.

Regardless of how you go about it, obviously there is going to need to be some protocol set for "rinsing" the anemone in a fresh SW bath (possible even saying use IO since it's cheap) then placed into a tank semi submerged and use sterile procedures (latex gloves and boiled scissors and hemostats/forceps could work for most hobbyists) to collect the sample. This could be placed inside a tube with say 5ml of fresh SW and sealed.

I would think using fresh SW in the tubes instead of dry or sterile saline may help to keep anything growing alive long enough for it to get to you. Once you receive it you could puree it or do whatever you needed to do to collect the information, but I feel developing something that is near impossible to screw up would be our best chance to ensure we're getting consistent samples.

I'm also with you Mihn in I feel that using statistics to "figure out" what's there that shouldn't be when comparing healthy vs sick nems would be the best way to go about this.

I'm hesitant with the mouth swap simply because I'm not sure we understand the anatomy of an anemone enough to ensure we're swabbing the proper parts to give us sufficient information every time. Too many things in there that all look the same for you, me and the next guy to all know we're swabbing the same thing.

I also feel that forcing deflation is a death sentence waiting to happen and I'm not positive that is the best option either.

[Amoo]

Kind of off topic here but kind of not so I'm going to post this here as a conversational side note.

The nem I got in yesterday went straight into a QT tank with a cycled HOB BioWheel filter. I usually keep a snail in these tanks just to have something to look at when there's nothing in an empty tank.

When I make the decision to turn the QT into a Tx I simply pull the HOB AND the snail and begin Cipro treatment. I then add water to a second tank and swap the Anemone back and forth between two Tx tanks while in treatment so I never have the nem out of water for more then 10 seconds or so during treatment.

Well this time I forgot to remove the snail. I'm about to clean the tank in the next hour, but from what I can see, he seems to have survived the 250mg Cipro treatment. I'll confirm this later today, but as I said I find it an interesting side about treatment.

[OrionN]

Thanks Amoo.
I am thinking of just aspirate 1 cc from the colum of the anemone. If he is infected, I would expect this aspiration would recover the organism (like do a blood culture in human) If we use aseptic technique, sterile needle and syringes, we should minimize the amount of contamination from outside of the anemone. What ever bacteria we recovered, it should be what inside the anemone.

I think this is what I will do. I will get several test healthy Gigantea to try this on and see how well they tolerate this procedure.

[raythepilot]

OK this project has really interested me. Have you tried to get a sample yet?

[OrionN]

I am working on logistic now. I need to get enough tanks set up and and light to keep these guys first.
I want to get 3-5 gigantea and Magnifica at a time. As soon as they arrive I will aspirate and culture them. Over the next day or two, depends on how they look, I will treat them. I will need to get enough system set up to house them first before I start anything.
For the purpose of obtain the information, I can just culture them and then flush them. Certainly this is not what I want to do, so I have to find accomodation for them first. My tanks are full. I will need to set up a system that will just keep these anemones. Right now I just don't have this set up yet.

Then not the least of which is how to get enough money for this. One set of culture will cost me about 30, more if I have to do drug sensitivity. The support equibments needed the keep them alive and well. I guess I can piggy these tank to my main system but each will need PH and light and circulation pumps.

So the bottom line is I am workingout the logistic now. I think I am reasonably sure about how to obtain culuter and will need to test on one or two anemone first to make sure that this will not kill them or serverely stress them.

[raythepilot]

I've read a few anemone research papers and they get around your problem by doing 100% water changes.
The pic enclosed is my holding tank. It holds only 1.7L of water. It sits inside another 10 gallon tank. I am treating the BTA in the holding tank with Cipro. I can add as much antibiotic as I like into the holding tank and not worry about messing up my tank biology.
I change the water in the holding tank 2x a day to make sure that the antibiotic concentration is always OK.
You can keep anemones in a tank like this, doing 1x 100% water changes per day, indefinitely. The trick is you have to be careful with your replacement water. My make up water is always 35.0 +/- 0.1 ppt and the temperature is +/- 0.2 deg C of the tank water.
It may be ugly hanging a bunch of these in your tank but it is easy, fast and effective.

[OrionN]

BTA is one thing. Magnifica and Gigantea is another. They are big, require a lot of water movement. I am not sure that air stone alone is enough water movement for these two species.
I have a 48X18 12H inches tank divided into 4 sections each 12X18 12H. This is my brood stock tank for my clownfish. I will use this to keep 4 anemones. I will use 4 PH for circulation, block off with egg crate so that the anemone cannot get to the PH. Now I just have to get enough light for them. I will have a huge sump with sand and rock for filtration. I thinking of just get 2 Radions for this and mount each Radion to light two sections. That should be enough light. Neither species require sand so I can have something that they will attach to that is easily remove.
So far that is what I am thinking.

[Amoo]

Could save yourself quite a bit of money and get a Photon or even one of the value fixtures without the auto dimming.

[CoralsAddiction]

BTA is one thing. Magnifica and Gigantea is another. They are big, require a lot of water movement. I am not sure that air stone alone is enough water movement for these two species.
I have a 48X18 12H inches tank divided into 4 sections each 12X18 12H. This is my brood stock tank for my clownfish. I will use this to keep 4 anemones. I will use 4 PH for circulation, block off with egg crate so that the anemone cannot get to the PH. Now I just have to get enough light for them. I will have a huge sump with sand and rock for filtration. I thinking of just get 2 Radions for this and mount each Radion to light two sections. That should be enough light. Neither species require sand so I can have something that they will attach to that is easily remove.
So far that is what I am thinking.


Given the fact that you like to see your fish early before work and after, having sunrise/sunset effect will probably be only beneficial to you. So I vote yes on Radions or any other light that has that feature.

[kris4647]

Minh has worked on the pathology and treatment of Mags and Gigs for many years. I respect his opinion immensely. Don't have time from the airport to Chime in with my experiences and ideas; but the methodology Dr Minh is espousing seems like a good jumping off point for experimentation. I have a few Petco 45 tanks Minh if you head my wAy ever.

[raythepilot]

BTA is one thing. Magnifica and Gigantea is another. They are big, require a lot of water movement. I am not sure that air stone alone is enough water movement for these two species.


I think you are confusing high shear with high water movement?
I can never turn my aerators full up. If I did in that little tank, the nem would spin like a towel in a clothes dryer!
The aerator has very low shear; so, the anemone can go right into the moving column. They often do that and if they stay in it, I start thinking that they are not feeling well.
The natural movement of anemone is down stream toward the stream intake. That is OK if it is an aerator, bad if it is a PH.

I was wondering, If you treat the nems in your tank with antibiotics, will that ruin your bio-filter?