Zoea syndrome


From: Anil Ghanekar
To: shrimp@yahoogroups.com
Sent: September 25, 2002


QUESTION:

Many hatcheries have reported high mortalities in
zoeal stages recently.
PCR tests show negative results for WSSV,
Higher levels of nutrition and higher densities in
Zoea have improved survival rates but the problem
still exists.
Any suggestions/solutions to this would be welcome.

Anil Ghanekar

e-mail: anilghanekar@yahoo.com

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COMMENTS 1:

You shouldn't have problems appearing with WSSV at larval or Zoea stages; this worry will happen latter.

As far as the Zoea syndrome has been understood in the Americas with vannamei, it may be an intracellular bacterial problem and with a very clear cross-contamination problem.

Then the best solution is to stock the hatchery at once (all the tanks) in the shortest time and then most (or all) of your hatchery tanks will make it. As soon as you reach mysis and latter stages you don't have this problem anymore.
So find a maturation hatchery that will provide you nauplii to stock in 2 or 3 days!

Eric Pinon

e-mail: epinon@ecua.net.ec

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COMMENTS 2 :

Some success has been reported by utilizing a cyanophyte (Aphanizomenon flos-aquae, AFA) at a 5% inclusion rate in the diet. There is ample publications supporting the fact that AFA causes the immediate release and circulation of N-K cells (natural killer). If you would like to investigate this further I would be happy to fax you the papers that reference this or you may read this on our website: http://www.desertlake.com

Howard W. Newman
Desert Lake Technologies, LLC

12750 Keno Worden Road
Keno, OR 97627, USA

e-mail: hwnewman@desertlake.com or bshimp@aol.com

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COMMENTS 3:

I think that Z2 problems have more to do with algae quality than with any other factor.  Most hatcheries find that by drying out after each cycle they can control or eliminate Z2, but no one has ever been able to show a good correlation between Z2 problems and any particular bacteria, virus or toxin.  Yet, when most hatcheries dry out, they also take their algae cultures back to an agar slant or refrigerator. Then they clean the algae section and start all over again with revitalized, clean cultures. Two or three weeks into larval production the algae quality is taking a nose dive and Z2 pops up. 

If you can get a good way to quantify "algae quality" (color, cell shape,
contamination, doubling time), I'll bet a strong correlation could be
established in most hatcheries with this problem.

Ed 

e-mail: edscura@aol.com

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COMMENTS 4 :

I think Z2 syndrome can affect animals at any stage and is a matter of how well balanced your system and controls are.  I have found that water quality, bacteria, poor nauplii, poor feed, culture environmental conditions and personnel can all contribute to difficulties in raising larvae. And you always have to consider broodstock nutrition as well if you are producing your own nauplii.  There are ways to test each one of the above, not to beyond a reasonable doubt but reasonably.

All aspects of culture need to be checked for quality, and cleanliness.
Bacterial work needs to be done on Artemia, algae, salt water supply,
freshwater supply, culture buildings and air supply and anywhere else you can think of.

You have some general things that all hatcheries need to do to minimize the effects of a Z2 syndrome.  Sterilize your water supplies by some means, we use chlorination using only 3 ppm because the water is so low in dissolved organics and because we have only one reservoir and I need to turn it around every 14 hours.  Your chlorination rate depends on how long you will have active chlorine in the water, I was once told 10 hours at low dosages should kill most things, then back it up with bacterial plates to make sure you are killing everything you want to.

Water lines need to be bleached at least once a week. Algae feed hoses or lines should be bleached at daily.  Artemia needs to be disinfected, alga should be started using antibiotic plates.

Residual chlorine may be undetectable because it can be bound by organics then released later affecting the organisms in the larvae tanks, too much thiosulfate can also affect you animals.

You need to look at the shrimp and ask yourself if you are seeing a water quality issue or a pathogen issue? Then take steps from there to test each possible scenario.

David Leong

e-mail: dleong@bluecadia.com

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COMMENTS 5:

We did a series of experiments using different water sources, including synthetics and did show a water quality impact on Z2 problems. Not enough resources available to pin down the problem, but it doesn’t appear to be any of the things we normally test for. But it was reproducible.

Dallas R. Weaver, PhD
Scientific Hatcheries
5542 Engineer Dr.
Huntington Beach, CA 92649, USA

e-mail: deweaver@gte.net

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COMMENTS 6 :

I agree water WILL make a difference in your survivals. The point I was trying to make is that it can be one or combinations of multiple factors contributing to Z2 syndrome, do not settle for a print out of what your water should and should not have, do not just listen to your managers saying that it is the water.  It is more than likely a combination of things affecting your animals. Deformities due to water quality and due to pathogens look different to me.  When I talk about Z2 syndrome I am referring to 40 to 90% of your larvae all dying at the same time whether it is at Z1, Z2, or even M1.

I have seen what I believe epoxy paint oxidation affect animals, 85%
mortality at Z1 with no EDTA, technician did not to add to tanks, this was for some reason corrected with 10ppm EDTA. Subsequent runs 50 to 60% survival harvesting at 75 to 100 animals per liter. The reason I guessed the tanks was because fibreglass 300L tanks stocked same day with same animals and no EDTA did 80% to Z3.

Less than perfect water may be acceptable for sustained production.  I do remember seeing a 10 % difference between 2 different artificial salts alone, don't remember where the hatchery water placed.  This would be a good place to start, perform a bioassay using artificial salts versus your properly sterilized normal source of culture water and check for a difference and make sure the results are reproducible. Then you may have either identified one of your problems or crossed one off your list. 

David Leong

e-mail: dleong@bluecadia.com

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COMMENTS 7:

Algae is a prime culprit. We are so lucky here in Fiji but I don't know how long we can keep it that way. We have bacterial and viral afflictions all around us but so far remain clean.

We all "whitewash" our hatcheries, clean up the whole thing and then go back to our stock cultures of algae. A few days later, we have a problem. I have experienced this in Australia and
East Africa. The so called "Z2" syndrome can hit you anywhere Z1, Z2, right thru to M3. Of course sanitize the hatchery completely, but remember to check the algae.

Alex Forbes

e-mail : docalec@connect.com.fj

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COMMENTS 8 :

So many possible reasons like water quality, algae, egg quality... Give a great attention to breeders nutrition (if maturation used) and select only the best spawns (check eggs and nauplii). You can also try to "wash" your nauplii in a specific tank, with a quite strong water exchange several years before entering into larval rearing ponds. 
If I remember well there are only one or two small shrimp farms in Fiji, right ?

Michel Autrand

127 Av. de Peyre Grosse
34980 Saint Clement de Riviere, France
tel/fax (33) 4 67 67 00 12
Mobile : 06 11 92 22 67
michel.autrand@wanadoo.fr
http://www.marine-cultures.com

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COMMENTS 9 :

We have all kinds of nasties around us in Fiji, but none in Fiji and it is our aim to keep it that way. Australia, New Caledonia, Indonesia, Philippines, they have it for the most part, yet here in Fiji, like the Seychelles many years ago, we are disease free. With frozen imports from Thailand and live imports, it is but a matter of time before we are subject to disease also.

Alec Forbes

e-mail: docalec@connect.com.fj

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COMMENTS 10 :

Here are some more details:

- This syndrome is recently observed in P. monodon hatcheries on the East coast, and a few on the west coast of India.
- Normal survival rates of 70 to 50% for last few years dropped to 40 to 20% in this cycle.
- Water treatment: 5ppm right up to 15 ppm chlorine, also some use only micron filtration and UV.
- Algae: Both Chaetoceros and Skeletonema are equally popular.
- No remarkably different bacterial loads seen in tank waters on TCBS plates.
- In some cases only about 20-30% tanks had extreme Zoea mortalities, while other tanks showed normal survival.
- Some reported smaller mysis size, and low mysis survival.
- In some cases, the problem started with lower algal blooms, but persisted, though to a lesser extent, even after the algae problem was corrected, or new algal strains were introduced.

Anil Ghanekar

e-mail: anilghanekar@yahoo.com

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COMMENTS 11:

We have not experienced anything we could call z2 syndrome, though we have had our problems and more! In conditioning pond-reared F. indicus, we find there is a conditioning process that affects the larval rearing success. Initially, survival from nauplius to Z1 is very low (20-30%), then as the broodstock condition, zoea survival improves.

Two years ago, an increasingly serious bacterial problem set in in out hatcheries and we could not get rid of it through dry-outs, sterilisation etc. We also tried a couple of probiotics and eventually found one that seemed to occupy the niche of our specific bacterial bug that affected us through all stages. This was related to pink stains on the floors of the LRT and often 90% zoea mortality in 10 hours. Such zoea would start off getting pale and pinkish and die rapidly. After the probiotic application, the problem basically disappeared. The carry-through has continued into this season, though we have not been able to afford the probiotic again yet (cannot sell last season's crop!). Without the probiotic, we have had our first instance of pink bacterial stains and related low performance in zoea-mysis in three years. Very subjective observation however.

As for our algae, we never "cleanup" using agar, but use the dilution method. Having read the comments on algae as a potential source of trouble, I am going to do a quick cleanup again. Basically, I stretch a pasteur pipette into a capillary under a flame and draw up a small volume from a mother culture using capillary action. I do this about 5 times, innoculating 5 different testubes with fresh L1 (Guillard's modified/updated F/2) medium. I then repeat this process from these tubes, so getting a massive dilution. I repeat this again, getting another massive dilution. Even a 4th dilution may be tried. These tubes are then left to grow. The very low dilutions may fail to grow, but weeks later, one or two will start to "smoke", showing algal growth. These are then checked microscopically and used for further propagation, or put through a second dilution process.

Additionally, we feed very healthy ("young") algae (Chaetoceros) to our zoea and add some nutrients to the LRT to encourage this algae to bloom in the LRT. We have fine tuned this to a routine: add the algae (100 litres) and 100ml L1 nutrients at 2pm the day before z1. This alga blooms to about 100,000 cells per ml by the next morning. This way, the zoea encounter healthy, exponentially growing algae.

Laurence Evans

e-mail : Laurence@amatikulu.co.za

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COMMENTS 12 :

I have technical experience when I was working in Colombia. We had several times z2 syndrome in one of our hatcheries. After following different ideas, we found down that water quality was the source of the problem.

We did different tests:

1.Grow shrimp with our first source of water (W1) and feed them with algae grown in the same water.

2.Grow shrimp with water from different sources (W2) and feed them with algae grown in the first source of water (W1).

3.Grow shrimp with water from different sources (W2) and feed them with algae grown in different sources of water (W2).

4. Grow shrimp with water from different sources (W2), and feed with poor quality of algae growing in different water sources (W2).

We used the same source of nauplii.

This test showed us that the problem was in our water because we got
Z2 syndrome in test number 1 and 2, but not in the test 3 and 4. We expected the syndrome in test number 4, but this never happened, also we got the highest survival in this test.

Now, working in USA with the environmental area, we are using plants and algae to clean polluted water. I could understand what happened, in the hatchery, years ago. I thought that any element in high concentration in the water could affect the normal animal behavior, as in some plants where iron is important for growth, but in high concentration it will be toxic.

When you have problems with algae it is because algae accumulate
some elements, which are present in high concentration in your water, and then they will be toxic for your animal. Sometimes, bacteria, virus, and fungi are not the first source in disease problems.

We changed our water source and some months later, we got the Z2  syndrome again, but just in two tanks, which came from different maturation source. We have never bought animal from this maturation, and then I ran the hatchery for 3 years. We were always working with high density; 250 and 300 animals per liters.

Robinson Bazurto

e-mail: buhocol@hotmail.com


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