[leebca]

FRESHWATER DIP WITH METHYLENE BLUE


INTRODUCTION
I totally support the use of a quarantine process for newly acquired marine specimens. My quarantine process and its benefits are written here: A Fish Quarantine Process
The matter isn't where the fish came from or how it was acquired. I don’t care how reliable the source (a best friend who guarantees his tanks and fishes have no disease; an LFS who says he did the quarantine for you; etc.) or how healthy you think the fish is (you watched it for days at your LFS and could see nothing wrong with it), it needs to be quarantined and before that quarantine, a freshwater dip.

Fish are always stressed. See: Stress (and the Single Marine Fish). But a proper quarantine process doesn’t have to be extra stressful to the new fish. The quarantine process can be made to be less stressful, if it is done properly with forethought. But, this post is not about the quarantine process. It is about what to do before the fish gets into your quarantine tank.

IMHO
All fish need to have a freshwater dip/bath before getting into your quarantine tank. It is essential for all fishes. There are just too many flukes, external parasites, gill parasites and things on and in newly acquired marine fishes that (especially to the eye of the excited aquarist who just bought the fish) are not readily visible. It is the greatest prophylactic step the aquarist can take to further protect their investment and the health of the newly acquired fish.

So here, I’ve provided as much (too much?) detail as possible about performing a good general freshwater dip for newly acquired marine fish.

THINGS YOU NEED

TWO identically sized plastic (non-clear) bowls such that (when equipped with the colander insert if you choose to use one) it is no less than 2 inches over the height of the fish and enough room for the fish to swim around in (or at least be able to easily turn around in).

ONE cover for the above bowls that is ‘open’ or has holes in it, like a plastic latticed cover or flat colander.

Methylene Blue (a solution sold for fish medication that is about 2.0 to 2.2% Methylene Blue)

Sodium Bicarbonate (Food Grade Arm & Hammer Baking Soda) to adjust pH up

Vinegar (pure without additives) to adjust pH down BUT avoid the use of this -- don't overshoot pH and prefer to dump the bath and start over rather than adding vinegar to the bath

Timer (stove, microwave, portable timer, watch, etc.)

Net(s) (preferably, use a colander (see below) to transfer and dip fish to avoid injury from a net)

Long plastic kitchen spoon (either solid or with holes) for mixing the bath

pH tester

Temperature tester


OPTIONAL THINGS YOU MAY WANT

A third plastic bowl (non-clear) exactly like the two from THINGS YOU NEED

Plastic Colander (that fits snugly into the above required plastic bowls; the fish will be inside this and it still needs to give the fish the 2" minimum water over the fish and swimming around room)

pH meter (Hanna handheld or similar model that measures to 0.05 pH units (or better) reliably)

Thermometer (easily portable digital that reads to 0.2 degrees F reliably)


PREPARATIONS

Prepare equipment:

There must be no sharp edges to the colander and plastic bowls (use fine sand paper then Emery Cloth to remove sharp edges)
There must be no labels or glue left from removed labels, on the equipment and tools.
Clean everything and rinse well with RO/DI or distilled water. (NOTE: Especially if you use a dishwasher for your household, you must rinse in hot tap water to remove any residue, then thoroughly rinse again in RO/DI).
Although it isn’t necessary, if you haven’t recently calibrated the pH meter (if you’ll be using one) I suggest doing that before this process.

Prepare the freshwater bath:

1. Use RO/DI water or distilled water for the bath water. Measure how much water you are using in gallons or liters.
2. Adjust bath water to about 3F above the temperature of the quarantine tank water (some cooling will occur). Stir with spoon.
3. Using Sodium Bicarbonate in small increments, adjust bath water to a pH just about 0.10 pH units below the pH of the quarantine tank water. Stir with spoon.
4. Add drops of Methylene Blue to the bath water as follows: 2 or 3 drops per liter of water; 9 drops (0.4 ml) per gallon of water. (NOTE: dispense drops holding the dropping bottle or dropper exactly vertical (not at an angle).)
5. Stir bath with spoon.
6. Finish adjusting the pH to match that of the quarantine tank water pH while stirring the bath.
NOTE: It is not impossible for the QT water to change pH in a short time. Be sure to verify that the bath pH still matches the pH of the QT water JUST BEFORE the fish is to enter the bath. Adjust pH just prior to use, if necessary.
7. Check temperature. If it is up to 2F above the quarantine tank water, that will be okay. If the bath water temperature is lower than the quarantine tank water, it must be raised. A lower bath water temperature than the quarantine tank water, is not acceptable. (You can use a microwave to heat a small portion of the bath water, then mix with the whole bath to raise the bath water temperature).

Prepare rinse bath:

Fill one of the bowls with quarantine tank water to use as a rinse, just before you need it (see DIP PROCESS)

----------------

So you have two (or three if using the colander method) identical sized bowls;

1 with the freshwater bath water in it, properly adjusted for pH and temperature
1 empty (to be filled when needed with quarantine water for the rinse off)
1 empty with the colander inside of it (if you're using nets, these two pieces aren't needed)

The colander will fit snugly into each of the three identical bowls


DIP PROCESS

Process directions are given as if you are using the colander suggested equipment.

1. Acclimate the fish in its bag, to the quarantine tank water according to: It Was Acclimation, I know. . .
2. After this acclimation and while fish is still in its transport bag: Gently pour the fish and water into the empty bowl with the colander insert in it. Cover the colander. (Now the fish is inside the plastic colander, which is inside the bowl, which now contains the bag water).
3. Lift the covered colander and let the water drain away out of the colander (and away from the fish). Hold for two seconds.
4. Insert covered colander into bowl with freshwater bath in it, and sink it.
5. Set timer for 5 minutes and start the timer.
6. For the first 5 minutes watch how the fish is doing through the lid or by peaking under the lid. Tangs often pretend to be dead. Don’t be fooled! If the fish is in legitimate danger, stop the bath and proceed to number 9.
7. After 5 minutes, check the fish again and start the timer again (for another 5 minutes).
8. Repeat 7. until the fish has been in the bath for 30 minutes or until the fish is in legitimate danger (see below: Signs of a fish in trouble).
9. Fill the still empty dry (third) bowl with quarantine tank water.
10 When time has come to stop the bath, raise the covered colander out of the bath, sink it into the rinse container of quarantine tank water to rinse off excess bath water. Pause just a few seconds.
11. Raise colander and sink it slowly into the quarantine tank so that the fish can quietly swim out.

Look closely at the freshwater bath water in the container. Do you see anything that came off the fish? It is maybe hard to see through the blue (so pour some off (decant it); dilute it with distilled water), but spend some time and look closely to see if anything came off. Look for anything the size and shape of a sesame seed (fluke); look for tiny specs the size of powdered sugar (Marine Velvet (Amyloodinium ocellatum)). This will help you with a diagnosis, should the fish end up being sick, infected or ?


TIME IN THE BATH

A freshwater bath experience is more stressful to the novice aquarist that it is to the fish. Most fishes can handle an hour in this bath, but 30 minutes is plenty. With experience you can tell the difference between a fish faking stress and one that is really in trouble. Hard to explain in writing, but I’ll try.

Signs of a fish in trouble. Often tangs (and a few other fishes) sink to the bottom of the bath and ‘play dead’ laying on their side. It is okay to prod it gently with a clean plastic rod or the long spoon handle, with no sharp edges. If it flops about or swims or swims then falls over ‘dead’ again, it is okay. Once some time goes by the tang will probably swim around. You’re mostly interested in how it’s breathing. Watch the gills.

Some fish will go into their defensive mode by erecting all spines, changing colors, changing patterns, or any combo of these. What the aquarist wants to do is don’t read too much into such antics, but to look closely at the fish to see how it is breathing. Rapid or normal breathing is okay for the first few minutes, but the methylene blue should calm the fish down shortly after the first few minutes (see below). If the fish is breathing very fast or not at all, with any one or more of those 'defensive' signs, the bath should stop.


BACKGROUND INFO

Without getting technical, the methylene blue helps calm the fish and provides the fish an easier time to breath. At the same time, the methylene blue will actually make life difficult for some external and gill parasites. It has a very nice dual use in this process so take advantage of it. DON’T overdose the bath water with methylene blue. More is not better!

I found you can find/buy inexpensive colanders and bowls from your 99 cent store, K-Mart, Wal-Mart, etc. You want a plastic colander that fits tightly into the bowls and gives the fish the most swimming/moving around room inside the colander. You can even buy such plastic containers from suppliers of aqua culture products through the Internet. Alternatively you can use nets, but as we all know, nets can harm the fish (and by the time you get the fish, the fish is totally afraid of nets!). Buy the bowl sizes that are right for the size fishes you will be dipping.

If you use nets rather than the colander method, then you don’t need one of those bowls and the colander. You’ll make the transfers using the net to move the fish from bath to rinse to QT.


SUMMARY

If you perform this process on all acquired fishes, you can save yourself a lot of grief of having to cure sick fishes, or even just give an acquired sick fish a good chance to fight off its affliction.

In the case of anemonefish: After they are in the QT they need to begin a treatment for Brooklynella. The freshwater dip will not clear them of Brooklynella. ALL ANEMONEFISH need to be treated for Brooklynella. It’s too common of a disease for these fishes not to do this treatment.

Similarly, after the fish is in the QT, all tangs of the Acanthurus genus should be treated with Cupramine copper treatment to rid them of both Marine Ich (Cryptocaryon irritans) and Marine Velvet (Amyloodinium ocellatum. These fish are so often infected with one, the other or both that treatment should be considered part of the QT process. I like the use of hyposalinity, but hyposalinity doesn’t kill Marine Velvet. Copper kills both these parasites.


More information you say?
Acclimation:
It Was Acclimation, I know. . .

Marine Fish Stress:
Stress (and the Single Marine Fish)

A fine quarantine article:
http://reefkeeping.com/issues/2004-1...ture/index.htm

Lee's Quarantine Process:
A Fish Quarantine Process

Now that the fish is in the QT, start feeding it right:
Feeding Marine Fish and Fish Nutrition

Lastly, if you find any 'things' in the dip water you are unsure about, post in this Forum. If you need any help with the QT process or a condition, injury or disease of your fish, post in the Marine Fish: Care, Health and Disease Treatment Forum here on Reefland.

Good luck!

[baobao]

Awesome post!! I just have a question on the length of time in this freswater dip - 30 minutes? is it necessary to go that long? wouldn't 10 minutes be sufficient?

[leebca]

Good question!

The dip is geared towards a variety of fishes with unknown conditions. The fishes with a thick mucous coat need the time for the freshwater to get to any organism that may be protected by this coat.

Totally exposed organisms (sitting on the surface in direct contact with the freshwater) can be eliminated in a few minutes. The traditional freshwater bath time was recommended for 1 to 3 minutes because in fact, the effects of surface organisms were seen in that time frame.

But when dealing with potentially 'protected unknowns' this procedure is the most effective. By this I mean that if I knew what organism I was dealing with, I'd administer the freshwater dip accordingly to succeed at affecting the target organism. Since the fish has been put through this, the 30 minutes v. 10 minutes doesn't mean that much to the fish in the way of short or long term negative impact. So. . .Safe and effective.

[Ixthys]

Wow! you weren't kidding about the Tangs! I just dipped 5 of them; the Bristletooth was the first and immediately went to the bottom and laid on his side. I noticed any time I looked into the bucket he was swimming around, but when he'd see me, he'd head straight down....what a player! The one that took it like a man (or man-fish?) was the Desjardini. Didn't flinch the entire time. At one point I think he even gave me the finger/fin!

Thanks for the heads up on this one!

[PufferDad]

What quantity of M-blue is to be used per gallon, liter? Almost lost my yellow tailed blue damsels in dip. How clear should the dip water be with the M-blue?

[leebca]

Part of the problem with giving directions as a number of drops is that a 'drop' can be of a wide range of sizes. In addition, you have to use the same concentration of Methylene Blue which the drop directions are given.

So, if the MB is in solution, that concentration of MB should be between 2.0 and 2.2% by weight, MB. If using this solution, the drop size is most uniform (and smallest) when the dropping bottle dispenser is held perfectly vertical when letting the drop fall. The number of drops recommended is based upon the MB bottle having a dropper cap on it. Held vertically, the number of drops per liter is 2 or 3 drops. Or, about 9 drops per gallon.

If it can be measured accurately (by calibrated or volumetric pipette), it would measure 0.4 ml of a 2.2% solution of MB, per gallon of water.

If you're looking through the side of a gallon container, the bath water would look a little darker than sky blue. The fish should be visible during the treatment, assuming the bath isn't more than a gallon or so in volume. The clarity of the water should be crystal clear. There should be no cloudiness or evident precipitation in the bath.

[bguile]

a few months back, before this thread, I tried FW dipping a Nox Angel and I believe an Auriga butterfly (memory eludes me) and I lost them both within a day or two. Granted I had not used the MB and brought pH up to the levels of the QT or display at the time. I floated the specimen container in the display water until the temp matched. Was there something totally wrong in this? Are there particular fish (other than sharks and the like) that should NOT be dipped like this? The FW dip and QT method have appealed to me in logic, but I'm afraid to risk another fish that I've just paid $40 and up for and it took me months to find it!

[leebca]

Probably the most important water quality feature in the dip bath is that the pH exactly match that of the QT water. Everything goes into the toilet if they don't match. The loss within a 'day or two' is pretty good evidence of an acclimation caused death. There are guidelines for that, here, too: It Was Acclimation, I know. . .

I just dipped a $400.00 fish using the above method. After dipping a few thousand fishes, it works, IF the directions are followed to the T. I focus on the lives saved (the fish is destined for a 300 gallon aquarium with more than $7,000 worth of fish in it!) rather than the possible loss of the new fish. This monetary value is added to the emotional value invested in the display tank fishes.

Good for all scaled fishes. If the care is followed about monitoring the fish's progress, then even the so-called 'delicate' fishes can be dipped and taken out of the bath before the full 30 minutes if and when they show signs of a negative reaction. Every fish has its own tolerance, but the vast majority can handle this dip procedure.

[bguile]

Sorry I'm just seeing the response to this one. Got caught up in the dialog of our other thread.

One question that has always concerned me was if we must acclimate a fish over time to a different salinity (ex. hyposalinity), then how/why is it ok to FW dip a fish? Why not just drop them into a QT where the salinity is kept at 1.009 or below?

[leebca]

That is a very good question. I always wonder why aquarists don't make that connection.

The answer has to do with how the fish handles the stress. Any change in salinity is stressful to the fish. If you take a glimpse at: Stress - A More Technical Discussion you'll note that the fish's reaction to stress is at first, physiological. There are chemical changes going on inside the fish to help the fish thwart or compensate for the stress. The fish need energy to do this.

A drop in salinity is not as significant as a rise in salinity. That is, the more stressful scenario is a fish trying to handle the stress of a rise in salinity. This is not meant to imply that there is no stress to the fish in dropping the salinity. There is still stress.

But the 30 minute drop and rise is short term. By the time the fish's internal biology ramps up the stress response chemistry, the fish is back in its 'normal' salinity. The fish is still stressed, only it is short term with some, but little affects.

If you take a fish and drop it immediately into a low salinity, say 11-12ppt, from a 33ppt salinity, the fish will lose stored energy over time in attempting to adjust to the sudden change. This is not good for a newly acquired fish which has most likely already lost much if not all of its stored energy. Taking 2 or 3 days to get to that salinity, will give the fish a transition to the new salinity, and time to gather more strength (eating the superior and nutritious food the smart aquarist is providing).

On the increase phase, a rise in salinity from 11-12ppt to 33ppt should take at least 7 days. The adjustment up is more stressful -- or read this to be -- the adjustment up takes more energy.

The dip is an acute event for 30 minutes. The sudden changing from 33ppt to 11-12ppt is a long term (chronic) event, taking a greater toll on the fish.

[WillieM96]

I've always wondered whether freshwater dips actually work. I've read "Freshwater dips are effective in killing free-swimming stages of amyloodinium; however, since encysted stages are protected, a single freshwater dip is not an effective treatment. Decreasing the salinity in a system has been suggested as a method for controlling amyloodinium epizootics, but because the organism flourishes in brackish water, the effectiveness of this strategy is doubtful." Am I missing something else not addressed here? I've seen this in several other studies but here's the link to this source: http://edis.ifas.ufl.edu/VM004

Thoughts?

[leebca]

Hey Willie! Thanks for posting.

That statement is 'mixed' up:

Freshwater dips are effective in killing free-swimming stages of Amyloodinium; . . .

The 'free-swimming stage' of Marine Velvet (Amyloodinium ocellatum) isn't on the fish. So it doesn't make sense that the dip (of the fish) would kill the free-swimming parasites that are in the water the fish comes from. See what I mean? The parasites IN the fish are called Trophonts. And the dip actually does kill many of those in the case of Marine Velvet. [Noga, 2000 and Noga & Levy, 1995]. Marine Velvet parasites are especially susceptible to FW dips when they have recently (less than 24 hours) attached themselves to the body of the fish.

I didn't check out the link you provided. If the above statement comes from there, and that bit about 'effective' and 'doubtful' comes from there (see below) I don't think it's worth my time to review it, IMO.

Once the Marine Velvet parasites gets into the fish, the fish protects it to some degree. Like Marine Ich (Cryptocaryon irritans) a freshwater dip won't kill ALL the ones buried in and protected by the fish, but it is often effective at ridding many of the parasites (Velvet and Ich) from the gill area. In the case of MV, these parasites in the skin of the fish are more susceptible to the affects of a FW dip (see above Noga references) than the MI parasites. The parasites attached to the gills are even more susceptible to being killed by a FW dip -- since the fish constantly bathes the gills with dip water, and the gills are not as heavily mucous coated.

MV likes and prefers to infect the fish in the gills first. By the time the aquarist sees signs of the disease on the body of the fish, the fish is not only heavily infected, but most likely to die. So whatever can be done to attack the parasite in the gills, where it: 1) is mostly likely hiding, and 2) is most susceptible, is a real advantage. That's where the FW dip comes into play for MV.

The dip does not cure a fish of either Marine Velvet (MV) nor Marine Ich (MI). It can help the fish fight off the disease if it has already begun, by reducing the parasites numbers on the gills (especially). The fish and aquarist could be lucky in that the FW could actually kill off all MV on the fish, but the odds are against it. But I hasten to add that the dipping of the fish isn't recommended because of MI nor MV.

If you read closely my recommendations. . .After the FW dip of Genus Acanthurus fishes, I still conduct a copper treatment to rid the fish of MI and MV.

In order of rationale, the purpose of the FW dip is to:
Rid the fish of external parasites (flukes, worms, etc.)
Rid the fish of gill parasites (flukes, worms, etc.)
Reduce the number or kill some other parasites (MI, MV, etc.)

Flukes, worms, and MI can be found in the dip water if the fish was infected with them and it is very good idea to inspect the dip water closely after the fish has gone through the process.

You'll find that one of the ways to identify a MV infection is to perform a fresh water dip and then to examine the dip water for MV parasites. It is more helpful to have a microscope to perform the examination, but a sharp eye can still detect MV parasites in the dip water. See: http://www.petsforum.com/personal/tr...fwdiptest.html

So, because I've seen the above work myself, there is no 'doubt' that the FW dip works on MV -- but not to cure it. So the part of the quote about the FW being ineffective I know to be false.

The last part about hyposalinity being a doubtful cure/treatment for MV. . .It isn't doubtful at all. It doesn't work.

[anthias]

yes I did it lee,
to day it was my first time to make afresh water dip for a maroon clown ,it took me time to adjuste PH value and temp ,but I matched it very precisely,the original water 8.2 so I set the qt and the fresh water 8.2
I watched the fish carefuly in the fresh water with my heart being fast but the fish seems to injoy it then I put the fish in the QT and now the fish looks great (12 hours in the QT) but I still have 44 days 12 hours to finish this QT process.
lee we still have to fined treatment for a syndrom caused with long time this hoppy takes,it is AWS (Angry wife syndrom)

[leebca]

Now there's a very luck fish!

[mhender]

Is it necessary to dip inverts and such? What about any macroalgae that is introduced into a tank?

/mh

[vaporize]

Originally Posted by mhender

Is it necessary to dip inverts and such? What about any macroalgae that is introduced into a tank?

/mh

Dont dip inverts, you'll kill them.

Macroalgae, you can do that but will kill the pods that might come with.

[leebca]

Like vaporize said, dipping invertebrates can be harmful. There are some exceptions, but being a 'fish person' I don't know them. I believe the anemone can be dipped but the procedure is different.

As far as invertebrates, live rock, macro algae, etc. are concerned, they are to be quarantined for sure, just no dipping unless there are instructions to the contrary.

[vaporize]

LEE, just a thought, I am always getting new fish into my systems (mostly clownfish that are going through different QTs). I am wondering if there is a fast way of storing FW dip water so I can just do a "match temperature and go" type of thing every time I get new fish in. Trying to adjust pH slowly with baking soda every time is killing me, takes 1/2 hr or longer in cases.

I tried to stored such RODI water + baking soda pH adjusted in food graded container. However when I use it later I found out that the pH actually dropped from the previously mixed 8.1-8.2 to 7.4-6. Any hints why? (maybe things settled)

Any tricks in storing this for constantly usage? (like once a week type of thing).

Thanks

[leebca]

I appreciate your challenge. I also appreciate the fact that you are doing what is right for the fish and your system -- giving a FW dip before release.

The FW dip doesn't lend itself to a continual, mass production or on-going type of application. The adjustment of fresh, pure water with Baking Soda is a temporary pH adjustment. Since the water is not buffered (like marine water) the FW with Baking Soda is unstable. As air gets to the water, there are chemical reactions going on which alter the pH of the dip water.

Part of the instructions note that just before use, the aquarist wants to verify the pH. The pH of the QT and the pH of the FW bath. pH can change in even the short time it took to prepare the FW dip.

Putting the dip water into an air-tight container would not likely work to hold the pH steady. The Baking Soda is releasing its own gas into the water which is going through changes to affect the pH.

I have no suggestions for long-term storage of dip water. Sorry.

Since I have come closest to your situation (in needing such water quite frequently) I have, more or less, gotten very good at quickly adjusting the pH and temperature of the water. I start off with water that is too cool. I microwave a bit of it to very hot. I slowly add this to the batch until the temperature is a degree F or two above my target. I make sure I have a known volume. Then, I put in some Baking Soda. I get a pH reading (a pH reading on pure water is almost useless -- there needs to be some ions present to yield a proper pH reading on a meter) and have learned within 0.1g how much more Baking Soda to add to bring the pH to the number I want. Remember: I use a constant source water and a constant volume every time I prepare the bath.

[vaporize]

Thanks LEE for the help, guess I just have to keep trying. FW dip + formalin bath for every new clownfish purchase takes ~ 2 hrs LOL I gotta find better use of my time hahhahahha

Learnt something new that I can show off to my friends now LOL My friend was always having a reading of 8-10 pH on his RODI water, so he asked me to find him an answer, I think I know why now, because the pH meter does not read any ions.