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non-viable embryos

non-viable embryos - Zebrafish Forum

non-viable embryos - Zebrafish Danio Rerio Forum. Discuss zebrafish, aquarium culture and molecular biology lab specimens.


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  #1  
Old 11-03-2010, 06:18 PM
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Default non-viable embryos



We are having problems with our embryos and I was wondering if you could possibly point me in the right direction.

The fishes we are using to breed are <1 year old (including the ones that you just sent us a few weeks ago) with most of them being between 6-8 months of age. Up until about a week ago, we were getting fairly decent clutches with normal development. Recently, though, the development of the embryos has been highly irregular. They are either laid as small and opaque, or they appear normal to the naked eye, but when they are placed under a microscope it becomes apparent that the cell division is highly uneven (ie one side is much larger than the other). This has been occurring on both of our systems (housed in the same room about 3 feet apart). Death occurs to approximately 90% of all embryos.

The water quality parameters have been averaging the following:

pH 6.9- 7.1
Conductivity 450- 520
Alkalinity 17.1 mg/L
Carbon Dioxide 8 mg/L
Cl ion 180 mg/L
Ammonia 0 mg/L
Oxygen 10 mg/L
Nitrite 0 mg/L
Hardness 119 mg/L (though this has been a little elevated lately)


I am doing a 10% water change daily, except for the weekends. To make my fishwater, I usually add about 175 ml of concentrated Instant Ocean (20 g/L) per 20L of ddWater. This varies a little depending on the conductivity measurement. Also, I am using the carbonite method of water buffering that we discussed previously.

As for feeding, I am feeding freshly hatched brineshrimp 2X per day (morning and afternoon) as well as feeding them with the Master Mix that ZIRC uses (I don't use the cyclopeeze in this, but otherwise it's identical to the recipe that you sent me).

Anyway, that's all that I can think of for right now. If you see any flaw in my protocol, I'd be open to suggestions.

Thanks in advance for all of your help
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Old 11-10-2010, 07:14 PM
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Default Re: non-viable embryos

Are you noticing any bacteria, fungi, or other parasites in the clutches when you look at them under a scope? Is the water you're using for the embryos just the same as the system water (I presume you're not taking it from the system, but is it made the same way)?

I always found it was a good idea to add methylene blue to the "egg water" - it acts as an antifungal and it's not bad for the fish. We would make 20L of egg water at a time and add a ml or so of methylene blue. The water will be fairly blue when you make it and will slowly photobleach. If it photobleaches to the point when you can barely see any blue color in the water, add more methylene blue.

Are you working with any other fish in the lab that have mutations which would cause early developmental defects? Is it possible there was a mix-up? That's the most common cause of odd phenotypes in fish labs (certainly far more likely than a mutation).

Cheers,
-Carlton
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Old 11-10-2010, 07:14 PM
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Default Re: non-viable embryos

One more thing... When is death occurring?
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Old 12-10-2010, 02:02 PM
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Default Re: non-viable embryos

Hi, I have the simmilar problem. Anything new with your embryos? Were you able to figure out something? Thank you,
ondrej
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Old 12-10-2010, 07:13 PM
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Default Re: non-viable embryos

Some dewormers (levamisole, specificaly) render zebrafish sterile. I don't know if it could also interfere with embryo development.
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