Laboratory Experiment with Popeye...

[jessibell]

I'm taking Microbiology for Animal Health Technologists at the moment, and a couple of my bettas developed popeye a few weeks ago and I decided to do some testing on it. So far the results are rather interesting so I thought I'd share them:

First I tested Melafix (diluted to the proper concentration for aquaium water) and a boiled down tea of Indian Almond leaves foreffectiveness against both a gram positive and gram negative bacteria. Both of them showed no evidence of being inhibitory (meaning they did nothing to slow the growth of the bacteria)

Next, I took water samples from an infected tank and a non-infected tank. Each of these water samples were from tanks that had a 100% water change exactly seven days prior. I streaked both of these on blood auger plates for isolated colonies and incubated them for 24 hours at 35 degrees C. (a little less than body temp.)

I was surprised today to see that the uninfected water had very little bacterial growth where as the infected tank had at least two distinguishable species of bacteria on it. I made slides of both of them and examined them under a microscope. They are both gram negative; the larger colonies showed hemolysis and are bascilli. the smaller colonies did not show hemolysis and are cocci.

My Microbiology teacher examined the colonies and states that they looked like enterobactia, which are normally found in the intestinal tract and can infect water through feces. We set up an entero tube for the larger of the two colonies which contains several indicators which should tell us what species it is.

Now that I have isolated colonies of these bacteria, I can set up some antimicrobial plates to test what antibiotics are effective against it.

It was interesting to me, as popeye is often caused by allowing the water to get too dirty (which I admit I did fail to do a weekly water change during mid term exams and this is when it happened ) so if it is indeed an enterobacteria causing the popeye, this would make sense. But the fact that the bacteria is still prevalent in the water after infecting the fish despite several water changes shows that once the infection takes hold, water changes alone are not going to get rid of it. I am now treating with kanamycin and salt and doing water changes every two days. I have heard ampicillin works against popeye, and we have ampicillin test disks at the lab that I can test it against. I wouldn't mind testing the kanamycin as well.

 

[mattrox]

Thank you for sharing this. It is great information.

 

[jessibell]

No problem, I am quite curious myself to identify the bacteria I have cultured from this tank - and either tomorow or next week I want to test both colonies against different antibiotics - starting with kanamycin, found in Seachem's Kanaplex, and a nitrofurazon/furazolidone complex (found in Jungle Fungus Eliminator) and, if I can get it, Maracyn 2 and Triple Sulfa. I can place test disks saturated in the reccomended dilutions of these drugs and measure the inhibition zone around them so see how effective they are. (basically, an entire petri plate is covered with one type of bacteria and these test disks are places on top, and if the drugs in them are effective against the bacteria, there will be a ring of "no growth" around them after incubating them for a day or two. The bigger the ring, the more effective the antibiotic is.

Of course, not all cases of popeye are bacterial - but the fact that I have a lot of funky growth in that tank compared to a tank with healthy fish seems like something is going on!

 

[mfdrookie516]

You lost me at "I'm"...

No seriously, you did! This is great info though. It would be cool to have some insight as to what causes diseases like this. While I'm sure you're not the first person to research this, I'll bet you're the first person from here I guess my question is, if not all cases of popeye are bacterial, then what exactly is it caused by then?

 

[jessibell]

Sometimes popeye can be caused by trauma or rough handling, and sometimes it can be caused by parasites

Yeah I have read all kinds of things about aquarium diseases in an attempt to find out more about it, but either the information is too vague to answer my questions or too scientific and convoluted. In microbiology we started to learn about bacteria and how they do their stuff and the more I learned about it, the more curious I got about my aquarium stuff. I'll try to keep in layman's terms as much as possible, as after all, I want the information to be a little useful if possible LOL

 

[ChizerBunoi]

That is very great and helpful information. Thank you for providing us with your test results.

 

[jessibell]

I should have the bacteria identified by monday or tuesday - and I will see about setting up some antimicrobial testing to see what works and what doesn't work.

 

[reefdiver]

**** your future job sound so cool.

 

[jessibell]

So we have the results back on the bacteria - they are not an enterobacteria like we thought. My micro teacher said it is either A Vibrio or Aeromonas species - which I found this interesting link on:

Aeromonas, Vibrio, Septicemia, Pop Eye: Treatment & aquarium diagnosis of.

These species can cause popeye, and commonly lesions. I don't see any red sores on my fish but the female is notably pale and shows a slight pink tinge around her gills - it was hard to tell if this was redness or just her pigment until she became so pale.

The site reccomends kanamycin/nitrofurazone which is what I am doing right now. It also says triple sulfa may work.

The only thing more we can do in the lab is do another water sample from the treated water and see if that has changed the bacterial growth at all. As for the fish themselves, they are not showing any improvements yet For those that are following along, do you think that raising the temperature would help knock the infection off or would it make it worse?

 

[WhiteDevil]

Popeye is a symptom of an interior bacterial infection, Id look at the fishes kidneys and other internal organs, I am sure you will find not only those two bacteria cells you and the professor found but you will find many many more.

"The affected fish should be immediately taken out to be separately treated. It is difficult to specify a specific treatment unless the main cause is definitely known. Large daily water changes should help, if not Epsom salts has been used with good results to draw the fluid out. One tablespoon per 5 gallons of water for at least three days, longer if necessary. Epsom salts isn't really salt (sodium chloride) it is Magnesium Sulfate. If water quality is the problem, a 50 percent water change must be made as soon as possible.

If a new item was added to the aquarium recently, it should be double checked that it is not poisoning the water or letting off chemicals. The quality of the water conditioner that is used should be checked that it has a good opinion by other aquarists. If the water readings are wrong (high nitrAtes etc), a 50 percent water change is recommended again and 15-20 percent water changes 3-5 times per week, until the water readings are correct. Overstocking is a common problem for high water readings. Another common reason would be decaying dead fish and fish food. The tank should be thoroughly searched for any dead fish and precautions must be taken NOT TO OVER FEED, and if the tank is overstocked, steps should be taken in finding some of your fish another good home. If a bacterial infection is the cause we would recommend 'Maracyn', 'Maracyn II', 'eSHa 2000' and 'Anti-internal bacteria' for treatment."