Our Second Issue

The format of our Guppy Labs e-bulletin may continue to evolve as it adapts to its environment. But what is for sure is that we are committed to developing a web-based platform to help disseminate accurate information about Poecilia reticulata. This is our second issue. Past issues of this e-bulletin will continue to be available in our archives.

Our first issue is now showing up when using the most common search engines. We are visited by readers from Singapore, Brazil, Germany, Indonesia, Taiwan, Thailand, Japan, Costa Rica, Canada, Spain, Uruguay, USA, Belgium, Estonia, Austria, Denmark, Australia, Poland, Malaysia, Indonesia, Italy, UK, Barbados, Vietnam, Mexico, The Netherlands, and more. Thanks to web sites like Guppyinfo.de, Guppy Club Singapore, Guppy-Forum.de, and Uruguay en El Acuarista, our first issue received 1,161 first time visitors and 2,165 hits during the first 90 days since publication. Not bad for the first issue! But then again, it's not only what you know that counts, it's also who you know! Thank you guys for putting a link to our e-bulletin on your web sites and discussion forums . We hope to continue to have your support. When it comes to the content of future issues, we promise to continue to strive for top-quality articles with relevant information. These first two issues are two firm steps in the right direction. Our only regret is that we are only publishing in English.

We hope you will agree that the information on this e-bulletin is relevant to the hobby. Coincidentally, this second issue has a tilt towards guppy nutrition. Three of the articles included in this issue relate to live feeds for guppies (Hsueh), types and performance of guppy feeds (Chaim), and vitamin C deficiency (Patiño); each providing a unique view of a practical example in guppy nutrition. This issue also provides an important historical account of guppy breeding and keeping in La Plata Basin, a border region between Uruguay and Argentina (Arijòn), and an important window into guppy keeping in Taiwan (Hsueh). We are also delighted to include an article about the beneficial uses of plants in the aquarium (Chris Ng).

If you are experienced with guppies, there is a lot to read between the lines. Hopefully that will motivate other to write articles for this bulletin. We hope someone from the Japan, Europe, USA, Canada, can contribute articles of interests or reports on the guppy scene for our next and future issues.

Enjoy reading and please e-mail this e-bulletin to guppy-people you know.

Hello to all my fellow guppy friends around the world. My name is Bruce Hsueh , and I am a guppy breeder from Taiwan. I think it is fair to say that breeders of Taiwan haven’t really been exposed to the rest of the world. When my good friend Tomoko asked me to write for Sergio’s e-bulletin, I just couldn’t pass by the opportunity.

There are many large scale breeders (200+ tanks) in Taiwan, and I have had the pleasure to meet almost all of them. I really have to admit, every one of them earned my admiration for their dedication and time spent in the field of breeding guppies. As large scale breeders, they often encounter many problems; Mother Nature, time, and cost efficiency. Among the cost efficiency, is the food for the guppies. How do you cut down the food bill, and be able to maintain fast growing and healthy guppies?

One of the breeders I had met had actually solved this problem by feeding his guppies live tubifex worms as their main diet. Yes, I know about feeding tubifex worms to your guppies, with the great risk, and all the hype about potentially and completely wiping out your fish room, virtually overnight. Also, how tubifex worms carry heavy metals, bacterial, protozoal, and nematode diseases, and should not be used as guppy food. All is true IF the worms are NOT processed, or cleaned, correctly.

From here, we will go to a breeder in Tainan (Nan means south in Chinese), and see how he has conquered this problem. His name is Felix Chen, and is the owner of South Bay Aquarium (breeds and sells guppies only), which is located on the second floor of a warehouse on the outskirts of Tainan City. Number of tanks he possesses: 720+. All his tanks are divided into three sections: breeders, baby and juvies, and grow-out. Babies and juvies are fed strictly with freshly hatched brineshrimp, and once they can be sexed, males and females are separated and moved into the grow-out tanks. All 480 tanks here are equipped with time-saving water exchange system. This is where they will do most of growing, by stuffing themselves silly with tubifex worms.

Here's the bottom line on feeding these creatures to your guppies. Nothing, and I'll say it again, absolutely nothing puts size on a guppy faster than regular feeding of tubifex worms. When you feed the tubifex worms, guppies attack and go into a feeding frenzy, as if they were possessed.

All tanks here are bare bottom, where worms can’t burrow and hide. Each tank has a double sponge filter, and one single tubifex feeder, where tubi’s are in constant supply.

On the corner of Felix’s warehouse, is a small room he has built, where he processes his tubifex worms. Ground level, and shaped in a square spiral cemented trench, is this ‘cleaning’ station. The clean well water (Yes, well water!) flows outwards from the middle of the spiral. See picture 1.

 

Picture 1 - tubifex worms process station

The width of the trench is approximately 8”, and built specifically to fit the rectangular baskets that hold the different batches of tubifex worms. The basket that is positioned closest to the water source, holds the cleanest batch of worms. It usually takes about five days to cycle up to that position. Hence, the basket that is positioned farthest away from the clean water source is the newly arrived and dirtiest batch. Once the cleanest batch is emptied and the basket has been removed, all of the other baskets will be moved one step closer to the clean water source. The reason for doing it this way is to prevent contamination, since the water only flows one way. To make sure the worms have enough oxygen, air stones are placed throughout the trench.

The source of these tubifex worms is from the local muddy and polluted streams, where people there sell them by the kilos. The price for one kilo of tubifex worms is about $90 NTD, or $2.50 USD for 2.2 pounds, in which toxins and parasites harbor in the stench surrounding these worms. According to Felix, it takes about three days to thoroughly “bleed” out their internal system, but to be on the safe side, and in accordance to his rigid standards, he will not use them for feed until they have been cleaned for five days.

After about five days on a “tubifex crash diet” for these worms, Felix gets about 75% rate of return by weight. The most amazing and noticeable difference is the smell. That awful, disgusting, and ‘puke-my-guts-out’ smell is magically…GONE! Felix relies heavily on his nose to judge the condition of a batch of worms. This is not recommended for anyone who has a weak stomach, or who has just eaten dinner.

Just exactly how safe is it? Felix claims that he hasn’t had any disease outbreak that is caused by tubifex worms, and has never had to de-worm his guppies, not even once! After three visits to his warehouse, I don’t recall ever seeing one sick tank. Some with bent spinal cords during the heat of the summer, but never a sick tank! Nothing devastates my pride and ego more than seeing another breeder who can raise bigger guppies than me. I had seen a DS male there, that at first glance appeared to be a 2” female, but upon closer inspection, turned out to be a not fully matured male.

Felix doesn’t hide the fact that he runs a successful business in South Bay Aquarium. His clients range from private collectors, to the aquarium stores throughout the Island of Taiwan, and even Hong Kong and China. One of the biggest reasons for his success… Feeding his guppies tubifex worms!

Here’s his website for those who knows how to read Chinese. http://www.guppy-taiwan.com.tw/

Picture 2 - Grow out tanks

Picture 3 - Felix Chen

Picture 4 - A newly arrived batch of worms

PLANTS IN GUPPY TANKS

In the breeding of fancy guppies, we often utilize bare tanks to facilitate the cleaning of mulm-fish waste, uneaten food (i.e. the stuff that is found on the bottom of the tanks).

Ammonia will be produced in the initial break down of the mulm and fishes do not take well to ammonia. Our guppies also produce invisible waste, which adds on to the ammonia load. The bacteria (nitrosomonas and nitrobacter) in established filters are able to break down the ammonia into nitrite and eventually into nitrate. Water changes are able to reduce the concentration of these nitrogenous compounds to a certain extent. Good water quality contributes to better growth and health in our fishes, enabling us to enjoy the hobby.

The aerobic bacteria can only break down ammonia and nitrite. Nitrate is present till a water change. We usually do partial water changes, removing only part of the nitrates. How can we reduce the concentration of nitrates further? A good method is the use of aquatic plants as we have lighting for our guppies too.

Benefits of aquatic plants

Plants take in nitrates readily for growth, especially for fast growing plants like duckweed and hydrilla. Plants also absorb carbon dioxide and produce oxygen during photosynthesis. By reducing the nutrients available to algae, less algae growth is present and incidence of green water is less.

Plants also provide hiding places. Over-zealous males might harass female guppies and plants provide a refuge from the attention of the males. Moreover, due to our hectic work, we might have missed out a pregnant female or a birth. Fries are able to hide in the plants and save themselves from becoming a meal for their parents.

Directly or indirectly, plants are a source of food for our fishes. Firstly, plants provides opportunities for the growth of micro-organisms such as infurosia. Secondly, as guppies are omnivorous, plant material is part of their complete diet. Allowing them to snack on the plants will definitely contribute to their well-being.

In our bare tanks, plants provide an aesthetic value. Plants come in various shades of green, red and brown, complementing the colours of our guppies.

Personal experience

All of my tanks/containers for guppies have plants… and most of them have no active filters.

Although none of my tanks can be considered as aquascapes, they are functional. The plants are growing, the fishes are active and growing well, the frequency of water change is reduced.

Some plants that I use and are found in some hobbyists’ tanks and guppy farms in my country

L: Amazon frog bits (Limnobium laevigatum) This is a fast-growing floating plant that absorbs lots of nitrates. It has long extensive roots which provides shelter for new-born fry and a host for micro-organisms. Algae tends to gather at the roots and removal of the alage-infested plants will suffice. Reaches around 3inches ( 7.5cm) in width.	R: Duck weed (Spirodela punctata) A tiny, rapid growing plant and troublesome to remove. I started with one plant and my tank was filled within a month! I do not recommend this plant in any guppy tanks.
L:Water hyacinth (Eichhornia crassipes) A big plant that produces beautiful purple flowers. Its root mass is a habitat of microorganisms that guppies could snack upon. However, it needs great light intensity and long duration. Massive nitrate sucker. Requires regular removal of dead leaves.	R: Java moss (Vesicularia dubyana) A good fry saver and requires little light. A slow grower that is a good habitat of micro-organisms, well suited for a fry tank. Mulm tends to gather beneath the moss.
L:Java fern (Microsorum pteropus 'Windelov') A shade loving plant that provides a haven for fry and micro-organisms alike. Easy to cultivate and aesthetically appealing.	R: Elodea (Egeria densa) The plant which I am holding. An easy to grow, fast growing plant, can be rooted or left floating. A great oxygenator and well-suited for a guppy tank.
L: Hornwort (Ceratophyllum demersum) A fast growing plant that has no roots. This plant produces a chemical that inhibits algae. Best left floating around in the tank.	R: Echinodorus 'Ozelot' A hybrid swordplant that grows up to 40 cm. Needs a deep substrate and has extensive roots. This plant requires lots of nutrients, mainly from the substrate; it can be grown emersed with its crown beneath the water surface.

In writing this article, I hope that fellow guppy hobbyists are able to beautify our tanks and bring out the beauty of our guppies further. These plants also contribute to better water quality at little cost to us, resulting in colourful and vigorous guppies.

If circumstances permit, I will follow up with an aquascaping article on the next issue of this E-bulletin.

Since I began breeding guppies, I have been concerned to feed then the best way I can, but without too many boring and expenses. At first I fed them only famous brand imported dry foods specially formulated for guppies, as many times per day as I could. Many times I saw brazilian commercially manufactured food-fish feeds work pretty well for food fishes. So I thought the same approach should work for guppies. I mean I trusted the labels... Also, these famous guppy feeds costs much more than food-fish feeds, so I thought this stuff should worthy. As I live, work, and breed my fish in the same property I run my business, I fed them every 2 hours. Perhaps you already know what happened.. After four months my juveniles were only middle sized and I was tired of stopping my work to feed fish.. I gave up! At least I learned that these aquarium-fish feed manufacturers do not know as much as what they charge for their products.

I went to the the common place of most breeders and I started feeding artemia nauplii to my guppies. Again, perhaps you already know what happened... The next generation grew bigger than the parents and were producing more fry than the grand-parents produced.

However I was not yet satisfied. I like to see my fish with a full belly. When feeding my fish a lot of bbs, the aquaria water becomes "bad" very quickly unless I returned to my every 2 hours feeding protocol, and I did not want to do so. Also, when looking for higher hatching outputs and/or money saving, I used low salinity hatching solutions which didn't allow me properly separate nauplii and shells. Last but not least was the usual wasting of some batches due bacterial growth when hatching cysts. Ok, I learned that artemia nauplii is fine for guppies but it is fine for bacteria too.

The next choice were moinas. I’m not sure if because they are freshwater beings, or if due to their reduced nutritional profile, but the quality of water in my aquaria improved (or didn't deteriorate as quickly). On the other hand, moinas fed on baker's yeast didn’t support the same growth rate like I had observed when feeding my fishes on bbs. So the logical step was to combine the “freshwater living quality” of the moinas to the nutritional profile of artemia nauplii. I then began to study about the live food enrichment methods, and about the nutritional compositions of moinas, artemia, beef heart and tubifex. From my lessons on enrichment methods, I got that since that newborn guppy fry could be readily fed on dry feeds. Plus it is easier to add the needed nutrients supplements to the dry feed instead to the live feed. In my case this new approach (moina plus home improved feed) worked better than artemia plus famous guppy feed, at least until the subsequent winter when I had to choose between to heat my aquaria or to heat moina cultures. I got back to artemia nauplii step.

In my searches on the nutritional profile of artemia nauplii, I found an article written by Garcia-Ortega et al. 1988 which brought me to the feeding protocol I'm using right now. They studied the factors which induce the superior performance of live organisms as feed for fish larvae, mapping the biochemical changes occurred in decapsulated artemia cysts during the hatching process. They also discussed their findings comparing the nutritional composition of decapsulated cysts and early stage nauplii.

On Table 1 we can see that protein and lipids increased, and carbohydrate and ash decreased significantly during cyst hatching when expressed as percentage of dry mater. On the other hand despite they found no significant differences there was a tendency for decreased concentration of all nutritional components per individual cysts/nauplii, mainly at later developmental stages where there was higher presence of nauplii in the samples. Although this may sound strange for you it is explained by a huge and disproportional decrease in individual dry weight observed during hatching. Let's say that you began the hatching process of 1 gram of decapsulated cysts dry matter, I said dry matter because water is not a nutrient, using the data in Table 1 we calculate you had in your hatching bottle 0.506g of protein (1g x 50.6%), and 0.147g of lipids (1g x 14.7%). After 24 hours, if ALL, I repeat ALL, cysts hatched into a nauplii, your fish feed shrunk around 32% in dry basis ( (3.42ug-2.31ug)/3.42ug ), these are the nutrients that embryos burned to grow and to leave their shells, so you will harvest ~0.680g of nauplii dry matter. By the end of hatching process you"ll catch in your nauplii net only ~0.382g of protein (0.680g x 56.2%), and ~0.116g (0.680g x 17%) of lipids, everything else is gone by your plumbing system to feed bacteria in the sewage treatment station, through your hands, fingers and pocket. Also there is another point related to the amount of nutrients caught per peck but it is highly variable as function of the different patterns of dispersion that nauplii may present in the aquaria, but if you assume that a fish expends the same amount of energy to catch a motionless prey (decapsulated cyst) or to catch a swimming prey (nauplii), and if you again assume that a fish catches the same number of individual prey items per peck, you'll realize that a fish eating decapsulated cysts ingests more nutrients per unit of energy expended to capture feed than a fish eating nauplii. Last but least, although in vitro protein digestibility data were not statistically analyzed we see a tendency for higher digestion rates of the protein from decapsulated cysts rather than nauplii protein.

Table from Garcia-Ortega et al. 1988.

Table 1. Proximate composition, individual dry weight and in vitro protein digestibility of Artemia at different developmental stages as function of incubation time

click here for larger picture

At Table 2 are shown the results for amino acid analysis. Among those 10 amino acids taken as essential for fishes 3 (threonine, valine and methionine) did not change significantly during hatching. Among the left 7 amino acids which changed significantly during development 5 (isoleucine, leucine,phenylalanine, lysine and tryptophan) were present in comparable amounts at both decapsulated cysts (1h development) and nauplii (25h development), just like total amino acid content. Finally, decapsulated cysts contained significantly more histidine and arginine than nauplii.

Table from Garcia-Ortega et al. 1988 .

Table 2. Amino acid composition (g 100 g-1 protein) of Artemia at different developmental stages as function of incubation time

click here for larger picture

At Table 3 are shown the results for fatty acids. Linoleic acid (18:2n-6), linolenic acid (18:3n-3), arachidonic acid (20:4n-6); eicosapentaenoic acid - EPA (20:5n-3) and docosahexaenoic acid DHA (22:6n-3) are taken as essential for one or other fish specie. Among these linoleic acid with cys configuration, linolenic acid, arachidonic acid and EPA changed significantly during incubation and in all cases were present in higher concentrations at decapsulated cysts rather than nauplii.

Table from Garcia-Ortega et al. 1988 .

Table 3. Fatty acid composition (mg g-1 dw) of Artemia at different developmental stages as function of incubation time

click here for larger picture

Garcia-Ortega et al. 1988 also studied proteolytic enzymes and they found no significant differences between cysts and nauplii. This last finding is very important for those thinking of feeding decapsulated cysts. Until the publication of this article, it was believed that superior performance of fish larvae/fry fed live feed when compared with ones fed man made diets was mainly due to the exogenous enzyme contribution to digestion. I should not say that exogenous enzymes do not play a role in digestion in all species because there are more enzymes involved in digestion than only the few proteolytic ones studied in this experiment, and because it was proven that supplementation of enzymes improved the survival and growth for some species . But for guppies, which are born with full developed digestive system and readily able to digest artificial diets, it looks that the exogenous enzyme contribution doesn't matter much (as suggested by another study cited below). The fact is that some fish species will grow better when fed decapsulated cysts instead artemia nauplii and some species won't.

Here in Brazil, we say that there are more things between the sky and the ground than could suppose our philosophy. But based on such data, Garcia-Ortega et al. 1988 concluded that “the superior performance of live food is probably more related to food intake and digestibility than to the biochemical composition of the food."

Well, at the time I found this Garcia-Ortega article, I was not interested on the use of decapsulated cysts, but to produce a feed matching the nutritional profile of the famous baby brine shrimp. It wasn't until I read a posting at Guppy Designer BBS stating that a breeder was feeding his fishes decapsulated cysts and he was producing the larger fishes he ever saw, that I got interested. At this time, feeding decapsulated cysts really caught my eyes...(If some day the person who posted that message comes across this article here, please, send me an email).

Last January, when I was at the library looking for something noteworthy to publish in this e-bulletin, I found a jewel from Singapore. It was an article by Lim et al. 2003, which brought me to another article, Lim et al. 2002, called “Use of decapsulated artemia cysts in ornamental fish culture.”

Dr. Lim and his partners compared the growth of guppy fry (1 day old) and adults (2 month old) fed dried decapsutaled cysts, brine preserved decapsulated cysts, Artemia nauplii or Moina. Following are the results of their study:

From Lim et al. 2002 .

In Table 1 from Lim et al. 2002 we see that authors found no significant differences in the stress resistance, wet weight, dry weight, total length and survival rate of the so called guppy “adults” when they were fed dried decapsulated cysts, brine preserved decapsulated cysts or Artemia nauplii. However, there was a tendency for guppies fed on dried cysts perform better than guppies fed on brine cysts and nauplii; I should say possibly due to their neutral buoyancy. Anyway, the guppies from all 3 groups did significantly better than the guppies fed on moinas which is the “standard” live food for guppy aquaculture in Singapore. In short, decapsulated cysts proved to be so good as nauplii as feed for juvenile guppys!

From Lim et al. 2002 .

In Table 2 from Lim et al. 2002, we see that results using guppy fry were different that result with guppy adults. Guppy fry fed dried cysts showed significantly better biometrics (wet weight, dry weight and total length) development than fry under all other treatments. Guppy fry fed brine preserved cysts shown somewhat inverse behaviour, with significantly better stress index, comparable survival rate but with significantly poorer growth than fishes fed dried cysts. Frys fed nauplii grew just like frys fed brine cysts but with somewhat lower welfare score than frys fed dried cysts. Finally moina eater frys performed a bit worst than nauplii eater's. About the intriguing low survival rates observed at this experiment, I should say they housed two hundred guppy fry in 40 liters of water (50 l. aquaria), and although they used box filters with aeration and they made 30% daily water changes, keeping water parameters at the “acceptable” range (temperature: 26-27.5C; pH: 6.4-6.6; dissolved oxygen: 7.5-7.9 mg/l; ammonia: <0.02 mg/l and nitrite <0.1 mg/l), and with no significant differences in these water quality parameters among treatments, I think that the low survival for all treatments is due to the rearing system adopted. Lim et al. 2002 also measured the ascorbic acid (vitamin C) and fatty acids contents in the feeds and fish tissue, for the different treatments. They observed that fishes which performed better were the ones fed feeds containing higher levels of n-3 highly unsaturated fatty acids, which is also know to reduce the effects of stress. interestingly, these feeds were relatively low in vitamin C, when vitamin C has been shown to boost resistance to stress in guppies.

Lim et al. 2002 concluded: “it became clear that the performance of fish fed decapsulated Artemia cysts was better than or similar to those fed Artemia nauplii or Moina”. Lim et al. 2002 highlighted the advantages of using decapsulated cysts as follows:

1 - More hygienic feed: All decapsulated cysts are disinfected by hypochlorite during decapsulation process. They are therefore more hygienic than potentially contaminated Moina.
2 - Off-the-shelf feed: Decapsulated cysts can be processed for a long-term storage, and may be fed to fish directly as and when required. This characteristic would ensure a ready supply of the feed to farms, and also the possibility of a more frequent food distribution to the culture tanks. The reduced feed retention time in the culture tank would lead to better water quality and lower risk of nutritional deterioration of the feeds.
3 - Lower Artemia cyst requirement: Decapsulated cysts constitute the highest energy form of Artemia and hence a lower amount of cysts would be required to provide same energy level as the nauplii. The improved energy balance of the decapsulated cysts may also result in substantial saving in feed cost.
4 – Labor saving: Compared with the use of Artemia nauplii, direct feeding of decapsulated cysts would alleviate the heavy work load in hatchery operation, as the labor intensive nauplii production is no longer necessary.

Last but not least, Lim et al. 2002 also discussed the economics of decapsulated cysts use. “It has been estimated that about 50% of the Artemia cysts have a low commercial value due to their low hatchability. These cysts can be acquired cheaply, at a fraction of the cost of the regular Artemia cysts (estimated to be US$ 6/kg vs. US$ 60/kg). After decapsulation and dehydratation the weight of decapsulated cysts is reduced by 46%, but this would not effect the utilizable amount of the cysts. Taking into consideration the cost of processing (US$ 12/kg) for decapsulated cysts and cost of hatching (US$ 6/kg) for regular cysts, the decapsulated cysts were estimated to cost 27% (US$ 18 vs. US$ 66 based on original weight of the cysts) of the regular Artemia cysts. For regular cysts, only a portion of cysts is hatched into nauplii for feeding, whereas for decapsulated cysts all the cysts can be used directly for feeding. When compared with an average hatch of cysts with 75% hatching percentage, the decapsulated cysts was estimated to cost 20% (27% x 75%) of artemia nauplii produced in the fish hatchery. As the dry weight ad energy content of decapsulated cysts are 30-40% higher than, for instance, 1 nauplii, the cost of decapsulated cysts is further reduced to 15% (20% divided by 1.35) of the Artemia nauplii in terms of feeding efficiency”. I didn’t understand all this explanation but I am quoting it just like it was published in the original article...

Yet, on the economical aspect, I would like cite : “Using decapsulated cysts as a direct food source for carp larvae, instead of nauplii, the quantity of cysts needed can be reduced by about 25% to 35% after one and two weeks of culturing, respectively.” Vanhecke et al. 1990.

When I wrote this article (march ‘04) Brine Shrimp Direct was selling 90%, 80% and 70% hatching cysts by US$25.95, US$17.95 and US$12.95/lb, respectively. On the other hand, they were asking US$13.95/lb for decapsulated cysts. Artemia International was asking US$13/500g of decapsulated cysts. I apologize for not mentioning prices from others suppliers, but I found no others price lists available in the web. Taking into consideration that one nauplii is around 2.4 micrograms in dry weight, that decapsulated cysts are 90% dry weight, that 1lb is around 454g, that hatching cost is zero and the prices and hatch-out rates of Brine Shrimp Direct products, you’ll see in the table below that the money saving as a result of feeding your fishes with decapsulated cysts instead nauplii is not so low as calculated by Lim at al. 2002, but it is yet quite significant when considering feed dry weight cost.

Product Hatch-out Nauplii/g Output              g dw/g of product Acquisition Cost US$/lb Acquisition Cost US$/g Final  Cost US$/g dw  Cysts 90% Hatch 245000 0.588 25.95 0.057 0.097 Cysts 80% Hatch 225000 0.540 17.95 0.040 0.073 Cysts 70% Hatch 195000 0.468 12.95 0.029 0.061 Decapsulated Cysts   0.900 13.95 0.031 0.034

I realize that the suppliers in the Artemia market prefer to sell decapsulated cysts (US$13-14/lb), but instead of non-hatching cysts (supposedly US$3/lb). I think that the prices for decapsulated cysts are still worth it. I suppose you have better things to do on your free time than decapsulating cysts yourself. Therefore, I won’t be too extensive on my explanation of the decapsulation process. But, if you are interested, I am citing the decapsulation processes described by Van Stappen in Lavens & Sorgeloos 1996 at worksheet 4.2.4, or by San Francisco Bay Brand or by Artemia-International or by Brine Shrimp Direct . I experimented with all methods and now I use this one described by Van Stappen .

Basically decapsulation process involves at least 3 steps: (1) hydratation, (2) decapsulation and (3) chlorine deactivation.

1 – Hydratation is did to make the cysts change from a “wilt ball shape” to a “full ball shape”, it is absolutely essential because it makes the cyst shell spherical and more readily removed;
2 – Decapsulation itself is the removal of the cysts shell external layers by the action of the hypochlorite solution. This reaction must take place at high pH (~9-10) and under 40C to preserve nutritional quality of embryos. I had observed that when decapsulating good quality cysts that the burning shell process is faster than for bad cysts which do not allow solution temperature raise above 30-35C.

Figure 4.2.1. Schematic diagram of the ultrastructure of an Artemia cyst. (modified from Morris and Afzelius, 1967)

From Van Stappen in Lavens & Sorgeloos 1996 .

3 – Chlorine deactivation can be made using sodium theosophies or acids like vinegar. I observed that when using acids occurred something like a coagulation of the cysts that makes me get afraid about protein denaturant which could decrease its protein digestibility. Now I neutralize chlorine by hugely washing cysts in tap water until chlorine smell is gone and dipping them in theosophies solution until chlorine test results colorless or negative.

Dehydration process could be omitted if you plan use all the cysts in less than a week. If so, they may be stored in the refrigerator using natural or artificial saltwater (Shyer, 2003). If you Decapsulated cysts for longer, the dehydration step becomes a must. I had observed that when cysts are dipped in saturated brine solution, they float and as they loose water they sink, they become darker and they also shrink in size. Right now I had an insight: I could try not dehydrating decapsulated cysts to preserve their buoyancy.Lim et al. 2002 concluded about the low survival rates observed in their study with molly and tetra fry fed brine preserved decapsulated cysts that “this was at least partly due to the rapid settling of the brine cysts, which made them less accessible to the pelagic fry during the first week of feeding. Hence more buoyant dried cysts should be used for feeding pelagic fish fry.” Somehow, that could also explain why guppy fry fed dried cysts grew better than those fed brine cysts for Lim et al. 2002.

About drying cysts, Lim et al. 2002 concluded in their study with guppy fry that “examination of the fatty acid profiles has revealed that dried cysts and guppy fry fed dried cysts are richer in (n-3) Highly Unsaturated Fatty Acids, including both EPA and DHA, than brine cysts and fish fed brine cysts respectively. The lower content in brine cysts could be due to their further metabolism after processing, as they were still alive, while the dried cysts were dead.” To produce dried cysts they washed brine preserved decapsulated cysts and dried them at 40C and forced ventilation until individual cysts were obtained.

Concluding I would like say that you to do not be afraid about feeding your guppies something relatively new, but yet scientifically proved to be worth it.

Thank you very much Dr. Jean Dhoti for allowing us access to the articles written by staff of the Artemia Reference Center – University of Gent– Belgium.

Further Reference:
Shyer, J. P. 2003. Live Feeds: Production of the brine shrimp Artemia without natural sea water or micro algae. Aquaculture Extension bulletin. University of Hawaii, Sea Grant Extension Service and UH CTAHR Cooperative Extension Service.

HIKARI Brand frozen baby brine shrimp

Can Nutritional Deficiencies Result in Bent Spines Under Particular Growing Conditions?

Right around December 20, 2003, one of my albino Coral Red double sword females had a drop of about 125 fry. Such large number of fry from an albino female ...I was very excited! I had brought this female all the way from Lund, Sweden; straight out of John T's fish room. This large drop would ensure my success in getting this strain established here in the USA, as long as they did well. Little did I know then what would be their fate...

Before I left for my trip to Scandinavia, I had been feeding frozen baby brine shrimp (bbs), which I had been hatching. But when I got back, since I had only a few fish to take care of and a bunch of store-credit at the local fish store, I decided to cash in and get a few packages of Hikari Brand frozen bbs.

Like I said, I had previously fed guppy fry my own frozen (newly-hatched) bbs exclusively with good results (i.e. no bent spines, and good growth and survival). I thought the Hikari Brand frozen bbs, being enriched with extra vitamins and such, had to be at least as good as my own frozen bbs. Was I right? I am not so sure. In fact, there are enough reasons to believe that the Hikari Brand frozen bbs used was deficient in something; perhaps vitamin C.

About 125 Newly-Born Albino Coral Red Double Sword Fry

Although you can't see it in this photo (above), these were normal fry when born, without any detectable abnormalities. Their mother was kept at 19-21 degrees Celsius for at least one month before birth. I took the fact that she had 125 fry all at once to mean that she had had adequate nutrition to that point. I also kept the fry in a shoe box at around 19-21 degrees Celsius, feeding Hikari Brand frozen bbs exclusively for 2 weeks. What happened during those two weeks is described below:

	You can clearly see the bent spines of these 2-week-old fry. Fry were reared at 19-22 degrees Celsius.
	These fry had adequate growth rate and their size was about right for their age. Other than being deformed (bent spines), these fry were eating well and not showing any signs of disease that I could see. Mortality was zero (0%) for this 2-week period.
	Almost every single fry in this batch had bent spines (about 90%). Obviously, this is not the result of high temperature. It has to be something else. It is also possible that there are two main types of deformities, but better photos and closer examination would be required to formulate a hypothesis about the types of deformities that resulted here.
	Even if seemingly straight when seen from above, some of the spines are bent backwards. Some of this fish appeared to have a "broken back", which was even more evident after they had eaten. You can see that these fry just ate. Their stomachs are (somewhat) full.

I had two other smaller batches of fry, one from a Japan Blue double sword female, and one from a snakeskin double sword female, which also developed the same abnormalities during that same general time period, but not in such a high rate (maybe only 50%). As soon as I realized the magnitude of this phenomenon, I realized what could be the problem. I changed diets and the problem went away. Now I think it had to be a nutritional problem. That is my hypothesis.

Under this hypothesis, the explanation for the observed deformities could be impaired collagen formation and/or decreased bone calcification. These symptoms are attributed to deficiencies of various nutrients, such as vitamin C (Sato et al., 1983; Soliman et al., 1986; Dabrowski et al., 1990) and minerals (Furuichi et al., 1997) in food, while deformities of rays were attributed to the deficiency of vitamin C and to the insufficient enrichment of live food with fatty acids (Gapasin et al., 1998; Gapasin and Duray, 2001).

What gives weight to my hypothesis is that, after changing diets, bent spines, were no more! Actually, I still see one or two here and there. As long as it is here and there, I can live with that. I've seen skeletal deformities occur at low rates when culturing other species . That is considered "normal", and there are several explanations for these occurrences.

Now I feed decapsulated brine shrimp cysts for a few days and then add mixed powdered flake/powdered trout starter in combination with decapsulated cysts for another few days. I am still hoping to figure out best feeding regimes for different size fry grown under semi-intensive conditions. I am looking for the right feed combinations for fry; something simple, inexpensive, and that works the best for me. The first 2-3 weeks of growth in guppies are crucial, and it is absolutely critical to get it right.

What About the Effect of High Temperature?

I've read in a few guppy web sites that high temperatures can result in fish with bent spines. I don't know of any scientific publication on temperature effects on guppy deformities. But there are several studies on the effect of temperature on skeletal deformities. Sfakianakis et al. (2004), reported a gradual increase in fish larvae (Pagellus erythrinus) with skeletal abnormalities as temperature increased, from 33% at 16 oC to 57%, 73% and 75% at 18, 21 and 23 oC, respectively. Temperature was also directly correlated with the severity of the deformities, expressed mainly as fish with more deformities simultaneously. Fuiman et al. (1998) concluded that higher temperatures accelerate the rate of development more than the rate of growth, thus resulting in different larval sizes at specific developmental stages. Environmental temperature has been shown to be an important factor for the development of anatomical abnormalities, especially at the extreme temperature and during the early embryonic and larval stages (Wiegand et al., 1989; Falk-Petersen and Hansen, 2001; Wang and Tsai, 2000).

Taking into account the effects of temperature on other fish species, it is possible that bent spines in guppies can result form exposure to high temperatures during the gestation period or during the first few weeks after birth. It would be useful to determine if the deformities are already present at the time of birth in guppies when their mothers are reared at high temperatures.

Perhaps someone should try supplementing diets for pregnant females and/of newly-born fry with 1000% excess in vitamin C and see if that helps counteract what may be the effects of high temperature on the development and growth of the skeleton in guppies. Maybe this would be a relatively simple follow up experiment: divide a batch of fry into two groups, rear them at the upper end of the temperature tolerance, and feed one group a diet without vitamin C and the other group a diet supplemented with 1000% vitamin C. Note: some manufacturers supplement their feeds with excess vitamin C. But be aware that vitamin C is not very stable and will deteriorate rather rapidly.

What About Genetic Factors (or Inheritance)?

Reports of genetic factors (inheritance) responsible for skeletal deformities in fish are rare (Divanach et al., 1996). Afonso et al. (2000) reported that a vertebral deformity consisting of a consecutive repetition of lordosis, scoliosis and kyphosis (LSK syndrome) could be genetically determined in Sparus aurata. Although it is highly unlikely that genetic factors were the cause of the phenomenon observed in the batch of Coral Red albino double sword guppy fry described above, the hypothesis of a genetic factor cannot be excluded. Especially when the degree or rate of deformities in that albino batch was higher than for two other batches of two different strains fed similar diet.

Looking For an Expert's Opinion

I decided to write Dr. Davis at the Department of Fisheries and Allied Aquacultures, Auburn University and ask for his opinion. We exchanged e-mail messages between March 28-30, 2004. Here is what he had to say:

GuppyLabs: Dr. Davis, I am looking for references regarding nutritional deficiencies causing morphological deformities in fish. Do you know of any relevant publications? Is there more to it that lack of vitamin C? Dr. Davis You are correct, vitamin C is the primary cause and is the most likely reason...If it is a large part of the population, then it is probably vitamin C deficiency. But bent spines can be caused by injury, deficiency of P (in combination with injury), etc. Halver and Hardy's new book on fish nutrition has a summary. The NRC (National Research Council) Nutrient Requirements of fish is much cheaper and also summarizes this. I do not know of a fish vitamin book per see but there is probably one out there. GuppyLabs Thanks Dr. Davis. This helps me a lot.... Dr. Davis What are you seeing this in (i.e. species, size, number, etc)? GuppyLabs Guppies. I had two batches (two strains) with ~50% and one albino batch with about 90%. I was feeding a commercial brand of frozen baby brine shrimp for two weeks after birth. I didn't noticed any deformities when newly born. But after about 2 weeks, these are the results. See pictures... Dr. Davis Vitamin C is very easily destroyed and will break down even in the freezer. I would suggest this is the primary culprit. GuppyLabs Thanks so much for your replies. There is a lot of anecdotal evidence in the international scene suggesting that high temperature also increases the incidence of bent spines in guppies. I am not exactly sure what sort of proportions we are talking about in those accounts. Would you guess vitamin C may be involved in that phenomenon as well, through some sort of accelerated metabolism/induced vitamin deficiency? Or could it be something totally unrelated? Dr. Davis You have several things at work at higher temperatures. Remember as the temperature increases, so does growth. Hence, the nutrients needed on a daily basis also increase. The daily intake for vitamin C, Phosphorus, as well as other nutrients required to build bone, cartilage, etc., all increase as the speed of the growth increases. The faster the growth, the more likely you will see deficiencies. Think of it like this if you are in you car on a flat road and you remove one spark plug you probably will survive the trip. However, if you are carrying a large load or going up hill, you will notice a major performance difference. My suggestion is that vitamin C is marginal at reduced temperatures (growth is reduce hence you are ok). However, as you increase temperature, you increases growth and you then have a deficiency. Just a thought. Good luck. I would try to get some prepared feeds (with vitamin's) into your guppies. GuppyLabs Dr. Davis, This is very helpful. I a writing a column for our GuppyLabs e-bulletin. With your permission, I would like to publish your comments there. This is what the piece will look like in our bulletin.. (bulletin.htm#hikari). It is in draft form. Dr. Davis Not a problem in posting my comments. However, I did not realize this was your goal. In the future you should make this clear. The note is quite good and will hopefully help other people. If you do not mind, I may use your notes as a practical example for my class. I have not really looked into nutrition of tropical fish but it would appear that there has been very little work done for this rather large industry. Good luck. GuppyLabs Yes, my bad. I am sorry!...Thank you! And yes, you can use this for your class. Dr. Davis Thanks, ...(The article) Looks great. Hopefully, you will keep a few people from making the same mistake.

Thank you Dr. Davis for allowing us to publish your comments

Comments From Hikari USA

On March 30th, 2004, we forwarded this article to Hikari USA (Hikari Customer Service <fish@hikariusa.com>). On March 31, 2004, Hikari replied with the following statement:

"Enrique - I will forward this information to our lab staff for their comments. Given the levels of stabilized vitamin C in this product, I have a strong suspicion it is not the food unless you have some of the variables suggested by the Davis staff. I will report back soon.

We did not hear from Hikari's lab staff after this initial contact with their customer service representative...

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References

Afonso, J.M., Montero, D., Robaina, L., Astorga, N., Izquierdo, M.S., Gines, R., 2000. Association of lordosis– scoliosis – kyphosis deformity in gilthead seabream (Sparus aurata) with family structure. Fish Physiol. Biochem. 22, 159–163.

Dabrowski, K., El-Fiky, N., Ko¨ck, G., Frigg, M., Wieser, W., 1990. Requirement of ascorbic acid and ascorbic sulfate in juvenile rainbow trout. Aquaculture 91, 317–337.

Divanach, P., Boglione, C., Menu, B., Koumoundouros, G., Kentouri, M., Cataudella, S., 1996. Abnormalities in finfish mariculture: an overview of the problem, causes and solutions. In: Chatain, B., Saroglia, M., Sweetman, J., Lavens, P. (Eds.), Seabass and seabream Culture: Problems and Prospects. European Aquaculture Society, Oostende, Belgium, pp. 45– 66.

Furuichi, M., Furusho, Y., Matsui, S., Kitajima, C., 1997. Essentiality of mineral mixture supplement to white fish meal diet for tiger puffer. J. Fac. Agric., Kyushu Univ. 42, 77– 85.

Gapasin, R.S.J., Duray, M.N., 2001. Effects of DHA-enriched live food on growth, survival and incidence of opercular deformities in milkfish (Chanos chanos). Aquaculture 193, 49–63.

Gapasin, R.S.J., Bombeo, R., Lavens, P., Sorgeloos, P., Nelis, H., 1998. Enrichment of live food with essential fatty acids and vitamin C: effects on milkfish (Chanos chanos) larval performance. Aquaculture 162, 269–286.

Sato, M., Kondo, T., Yoshinaka, R., Ikeda, S., 1983. Effect of water temperature on the skeletal deformity in ascorbic-acid deficient rainbow trout. Bull. Jpn. Soc. Sci. Fish. 49, 443– 446.

Soliman, A.K., Jauncey, K., Roberts, R.H., 1986. The effect of varying forms of dietary ascorbic acid on the nutrition of juvenile tilapias (Oreochromis niloticus). Aquaculture 52, 1 –10.

Wang, L.-H., Tsai, C.-L., 2000. Effects of temperature on the deformity and sex differentiation of tilapia, Oreochromis mossammbicus. J. Exp. Zool. 286, 534–537.

Wiegand, M.D., Hataley, J.M., Kitchen, C.L., Buchanan, L.G., 1989. Induction of developmental abnormalities in larval goldfish, Carassius auratus L., under cool incubation conditions. J. Fish Biol. 35, 85– 95.

LEBISTES OR GUPPY

The Full Blue of the Río de la Plata

Part One

Argentina with thirty three million inhabitants and Uruguay with three million coexist at the south of South America sharing deep popular roots such as tango, pies, dulce de leche, asado and the mate. Tango Empanadas Dulce de leche Asado Mate The liking for aquarism aroused simultaneously in both borders of the Río de la Plata. There has been almost no diffusion of our activity abroad. Internet provides the means to fill the gap between past and present. Regarding lebistes, we have updated our knowledge due to the fact that we have exchanged opinions with foreign breeders taking into consideration differences and similarities. During the last 20 years, Uruguay has been breeding a lebistes with colour on the upper part of the head and a basic blue tonality from the snout to the caudal fin. These variations developed in Uruguay were called Full Blue. They present different degrees of dull, metallic or iridescent colours. Some variations have darker areas in the front body and around the pectoral fins. Another characteristic shared by almost all the Full Blues is the major or minor motility of their pigment, which does to a temperamental coloration fish. In Uruguay, we found out about the origin of this mutation because Carlos Pastre brought back individuals with such characteristic pattern during one of his visits to the breeding place of Professor Daniel Tejedor in La Plata, Argentina. Carlos Pastre bred these fish and their descendants showed a deep dark blue colour from the snout to the outer edge of the caudal fin. During a trip to Buenos Aires, I purchased three males in the Fair of Pompeya, descendants of Tejedor´s stock. They had full greenish bodies, heads and fins, with white outer borders of the caudal fins. From these three males, I developed Full Blue variations with solid and fantasy caudal fins. The caudal fins that present marks or spots of diverse colours and sizes name the uruguayan “Fantasy Lebistes”, while in other countries the uruguayan Fantasy pattern is called Glass, Grass or Mosaic. Photos by Rosario Arijón Juveniles Analyzing differences and similarities, I found doubtless resemblance between the Full Blue lebistes developed in Uruguay and the characteristic pattern of the famous Moscow guppy, highly appreciated all over the world. I contacted Professor Daniel Tejedor, whom I have met years ago during one of his trips to Montevideo where he visited my fish room, in order to understand the origin of this type of fish. Professor Daniel Tejedor is licensed in Zoology (CsNs), doctorate with thesis in genetic markers, teacher of genetics for twenty years in the University of La Plata with exclusive dedication (investigation and teaching). He has done aquiculture fieldwork, has taken courses in Japan and has been teaching in Ciencias Veterinarias in the University of Tandil for four years. Tejedor, "old lebistero" according to his own definition, owns a fish farm of 25 hectares in Oliden, with 30 diggings, more than two hectares of water, thirty thousand cubic meters, two supplying pumps with a total 200 cubic meters per hour. Fifty kilometres away is the family breeding place where he works with a water volume of 150.000 to 200.000 liters, with a pump of 7.000 liters. Besides guppys, he breeds platys (mickey mouse, red, tuxedo and black strains), swordtails (tuxedo, red, berlin, black, neon and high fin lyretail strains), koi, carassius (comet, shubunkin, black, goldfish and calico strains), herbivorous carps and Blue Jack Dempsey. Romina Tironi, who’s in charge of fish keeping and breeder’s selective care has been working in Tejedor’s establishment for the past nine years in order to afford her odontologist course. She pays attention to any variation that might occur such as the albino guppys that came from one unique albino female found by chance. Some of Tejedor 's guppy lines have more than thirty years for shape and colour selective breeding. His guppys are perfectly adapted to breed at 18ºC and live at 14ºC. When asked about the body pattern in question, Professor Tejedor stated that it is just one more of the many mutations occurred in his establishment along his fish breeder life. It was about 1970 when two males and three females 3/4 black, of excellent quality, originated his Multicolors, 3/4 Blues and the Full Blues. Those five 3/4 Black guppys, ancestors of Full Blue, were a gift of his friend Alfio Ramina. Alfio Ramina got that original black line from Andrés Randazo who created those 3/4 Black guppys that produced a percentage of recessive Half Black Blue. Dr. Oscar de Barrio recalls that his friend Andrés Randazo created 3/4 Blacks by crossing Harlequins and 3/4 Blues. The Argentinean Harlequins are as the Uruguayan Africans called Red Tuxedo or Half Black Red in the rest of the world. After exchanging information via emails with Professor Tejedor, we shared thorough