JPH0336486B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an artificial culture method for shrimp and crabs, which is capable of keeping the mortality rate as low as possible during the metamorphosis and molting stages of shrimp and crab growth. The purpose is to provide aquaculture methods.

In recent years, many attempts have been made to artificially farm shrimp and crabs, which are in high demand and expensive despite the declining harvest, but artificial farming methods have been established that are profitable due to the high mortality rate. The reality is that it is difficult to say so.

As is well known, shrimp and crabs undergo metamorphosis and molting during their growth process to grow into adults.

For example, the fertilized egg of a spiny lobster passes through the Nauplius stage while being attached and protected to the mother's abdomen and swimming limbs, and after about a month it becomes a phyllosoma and enters the Zoea stage. As they live a floating life, they repeatedly shed their skin and grow larger, becoming Mysis.
During this period, they molt once and become adults.

Incidentally, American rhinoceros shrimp, long-eared shrimp, and black shrimp also become adults through the same growth process as described above (however, black shrimp are laid eggs in water, so they pass through the naupuri period in water).

In addition, for example, the fertilized eggs of a freshwater crab go through the naupli stage while being protected by attaching to the abdomen and appendages of the mother's body, hatch after about 1 to 3 weeks, become independent from the mother's body, and enter the zoia stage. During this period, the body grows larger as it moves its abdomen back and forth and floats around, repeatedly shedding its skin and becoming a megadonkey (Megalopa).
During this period, they molt once and become adults.

Note that other types of crabs also become adults through the same growth process as described above.

When artificially cultivating shrimp and crabs that follow the growth process described above, the mortality rate during the metamorphosis and molting stages is extremely high.

For example, the case of artificial cultivation of prawns is as follows.

When fertilized eggs released from prawns with ripe eggs are collected using a net and placed in a culture tank for cultivation, the survival rate during the naupuri stage is as high as approximately 90%, as they live on the egg yolk in their bodies. During the zoia stage, the survival rate when fed with diatoms is very low at about 28%, and when the mysis stage is reached, the survival rate increases to about 86% when fed with zooplankton.

The same applies to other types of shrimp and crabs, and the mortality rate during the zoia stage is extremely high.

In view of the current situation as described above, the present inventors
Systematic research has been carried out for many years in search of artificial aquaculture methods that can keep the mortality rate of crabs as low as possible during the growth process, but in the process, artificial aquaculture of shrimp and crabs has been carried out for many years. The commonly used compounded feed contains photosynthetic bacteria such as purple nonsulfur bacteria, purple sulfur bacteria, and trace amounts of vitamins.
When feeding with _B1 , para-aminobenzoic acid, nicotinic acid, and biotinic acid, the mortality rate can be extremely low and growth retardation or stoppage can be prevented. I got the necessary knowledge.

That is, in the artificial cultivation of shrimp and crabs, the present invention adds purple non-sulfur bacteria and/or purple sulfur bacteria and a trace amount of vitamin B ₁ to commercially available compounded feed used for artificial cultivation of shrimp and crabs. , para-aminobenzoic acid, nicotinic acid, and biotin are added to the shrimp or crab, thereby reducing the mortality rate of the shrimp or crab during metamorphosis and molting. There is an artificial cultivation method.

The structure and effects of the method of the present invention will now be explained as follows. First, I will explain in detail about purple non-sulfur bacteria and purple sulfur bacteria. Both of these bacteria are well known as photosynthetic bacteria and live in most places with standing water in the tropics and subtropics (paddy fields, ditches, sewage, rivers, etc.), and the details listed below are also well known. .

Purple non-sulfur bacterium - Asiorodase:
Athiorhodaceae has the following genera (one representative species belonging to each genus is listed and its mycological properties are described).

1 Rhodopseudomonas
Genus Capsulatus: Capsulatus species a Morphological characteristics Extremely motile with a single flagellum;
Usually, it is a short rod-shaped bacterium (width 0.5μ x length 1.0μ), but depending on the type of culture medium and culture period, long rod-shaped bacteria (width 0.5-0.7μ x length 6.0μ) may be produced. It comes out. That is, it shows a polymorphic phenomenon.

b Conditions Growth conditions in various growth media (under anaerobic lighting conditions) Meat juice + peptone water +++ Potato medium − Thiosulfate − Alanine + Leucine − Asparagine + Aspartic acid − Glutamic acid + Tartaric acid − Citric acid − Glutaric acid + Acetic acid + Propionic acid +++ Lactic acid ++ Succinic acid + Malic acid + Butyric acid ++ Crotonic acid + Pyruvate ++ Ethanol - Mannitol - Sorbitol - Glucose + Mannose - Fructose + Glycerol - (both use 0.2% concentration for substrate) Note: +++ → Good growth + → Possible to grow − → Unable to grow c Physiological properties (1) Optimal growth conditions pH 7.2, temperature 27℃, anaerobic lighting (10000 lux) (2) Conditions for growth pH 6.0 to 8.5 temperature 23 ~39℃, aerobic to anaerobic dark conditions ~ lighting conditions (3) Gram staining negative (4) Acid-fast (5) Indole production None (6) Hydrogen sulfide production None (7) Ability to fix nitrogen gas Yes (8) Nitrate media also perform denitrification, which is the opposite of nitrogen fixation by reducing nitric acid and gasifying N ₂ ↑.

(9) Catalase production Yes (10) Gelatin liquefaction None (11) Starch hydrolysis None (12) Oxidation ability of reduced methylene blue, reduced methyl (or benzyl) biolodiene dyes (13) Biotein, thiamiin, Requires nicotinic acid as a growth factor.

2 Rhodospirillum: Rhodospillum genus Rubrum: Rubrum species a Morphological characteristics The initial culture is a flagellated, motile spiral-shaped bacterium (0.5-1.5μ x length 2.0-5.0μ). Shows polymorphism.

b Growth conditions Growth conditions in various media (anaerobic lighting conditions) Meat juice + peptone water + potato medium - thiosulfate - alanine + lyucine ± asparagine + aspartic acid + glutamic acid + tartaric acid - citric acid - glutaric acid - acetic acid + Propionic acid + lactic acid + succinic acid + malic acid + butyric acid + crotonic acid + pyruvic acid + ethanol + mannitol - sorbitol - glucose ± mannose - fructose - glycerol - (both use 0.2% concentration for substrate) Note: + →Can grow ±→Grows or does not grow −→Cannot grow c Physiological properties (1) Optimal growth conditions pH 6.8 to 7.0, 30 to 35℃ (2) Conditions for growth pH 6.0 to 8.5 favorable Aerobic to Anaerobic Dark to Light (3) Gram staining Negative (4) Anti-acidity Yes (5) Indole formation None (6) Hydrogen sulfide formation None (7) Ability to fix nitrogen gas Yes (8) Nitrate The culture medium also performs denitrification, which is the opposite of nitrogen fixation, by reducing nitric acid and gasifying N ₂ ↑.

(9) Catalase production Yes (10) Gelatin liquefaction None (11) Starch hydrolysis None (12) Reduced methylene blue, reduced methyl (or benzyl) biolodiene oxidation ability Yes (13) Biotin as a growth factor request.

Purple sulfur bacteria - Siolodase:
Thiorhodaceae has the following genera (in addition,
One representative species was listed and its mycological properties were described. ).

1 Chromatium: Chromatium genus Vinosum: Vinosum species a Usually oval to short rod-shaped (width 1-4μ, length 2
~10 μ) and motile due to flagella, b Growth conditions Growth conditions in various media (anaerobic lighting conditions) Meat juice − Peptone water ± Potato medium − Thiosulfate + Sodium propionate − Sodium malate + Sodium succinate + Glucose - Etonol - (Both substrates used at 0.2% concentration) Note: + → Viable - → Unviable c Physiological characteristics (1) Optimal growth conditions (pH 8.2, temperature 30℃, anaerobic light (10.000lux) )) (2) Conditions for growth (PH7.6-8.8, temperature 25℃)
(~40℃, anaerobic lighting conditions) (3) Gram staining is negative (4) Acid-fastness No remainder (5) Indole formation None (6) Utilizes hydrogen sulfide very well.

(7) Nitrogen fixation ability Yes (8) Does not grow on nitrate medium (9) Catalase production Yes (10) Gelatin liquefaction No (11) Starch hydrolysis No (12) Reduced methyl (or benzyl) biologien dye Perform oxidation.

(13) Vitamin requirement None The methods for harvesting the purple non-sulfur bacteria and purple sulfur bacteria listed above have already been established. It is well known that a large amount of bacterial cells can be obtained during the purification process of starch production waste liquid, etc.). Furthermore, various culture methods are known as mass culture methods, and a typical example thereof is as follows.

That is, in the nutrient source mixing tank (NH ₄ ) ₂ SO ₄ ,
KH ₂ PO ₄ , MgSO ₄・7H ₂ O, NaCl, CaCl ₂ ,
When culturing purple sulfur bacteria by mixing and dissolving culture media such as NaHCO ₃ and yeast extract to create a basal medium, add Na ₂ S and 9H ₂ O to this basal medium to make a culture medium, and create a purple sulfur-free culture medium. When culturing bacteria, lower fatty acids such as acetic acid, propionic acid, and lactic acid are added as Na salts to this basic medium to create a culture solution. In either case, this culture solution is transferred to a closed, illuminated culture tank, and The purple sulfur bacteria are transplanted and cultured.

To explain more specifically, the culture medium composition is 0.3 g of (NH ₄ ) ₂ SO ₄ per 1 part of tap water;
_{K2HPO4 0.5g} , _MgSO4・_7H2O0.2g , NaCl0.5
g, _CaCl2 0.5g, _NaHCO3 0.2g, yeast extract
In the case of purple sulfur bacteria, use a basal medium containing 0.01g of Na ₂ S・9H ₂ O.
Add to 0.5% and adjust pH to 8.2-8.5 with KOH solution.
In the case of purple nonsulfur bacteria, add 0.3 to 0.5% of lower fatty acids such as acetic acid, butyric acid, and lactic acid to the above-mentioned basal medium to prepare a culture solution. In this case, the pH is 7.

If the above culture solution is placed in a transparent container such as glass, sealed, and cultured for 2 days at 30°C under 5000 lux lighting, approximately 3 g of bacterial cells can be collected.

The general components of the bacterial cells harvested by the above culture method are 55.4% protein, 2.3% fat, 25% sugar, and fiber.
2.9%, ash 4.4%, and vitamin A content.
864600I.U./100g, vitamin C is 161.2mg%.

In addition, the content of vitamin _B12 reaches 200γ% (incidentally, yeast has only 1γ% and chlorella has only 10γ%). Furthermore, the content of carotene pigments (lycopene, rhodopine, spirilloxanitin, etc.) and bacterial chlorophyll is extremely high, reaching 1000 to 3000 mg%.

In the method of the present invention, when using purple non-sulfur bacteria and/or purple sulfur bacteria, wet bacterial cells or freeze-dried bacterial cells may be used. In the former case, it is convenient to add it to the target commercially available compounded feed by spraying.

The amount added must be at least 0.1% by weight or more based on the commercially available feed, and if it is less than this, it is difficult to obtain a significant effect.
The upper limit is not particularly limited, but in practice it is
Addition of about 10% by weight is sufficient.

Next, as seen in the examples below, a sufficient effect can be obtained by adding purple non-sulfur bacteria and/or purple sulfur bacteria, but in addition, a trace amount of vitamin B ₁ ,
Greater effects can be obtained by adding para-aminobenzoic acid, nicotinic acid, and biotin.

The amounts of the above vitamins added may be extremely small;
A few micrograms to several tens of milligrams per 100 grams of commercial feed is sufficient.

The reason why the effect is significantly improved when purple non-sulfur bacteria, purple sulfur bacteria and the above vitamins are used together compared to when they are not used together has not yet been theoretically elucidated. However, it has been confirmed that adding the above vitamins to commercially available mixed feed has almost no effect.From this point of view, the above vitamins do not inhibit the growth of purple non-sulfur bacteria and purple sulfur bacteria. This is considered to be a promotion.

Next, we will discuss commercially available mixed feeds used in artificial culture of shrimp and crabs.

Generally commercially available compound feed for artificial culture of shrimp and crab is a cylindrical product with a diameter of about 3 mm and a length of about 15 mm, and its composition is 35 to 45% crude protein and 2 to 20% crude fat.
5%, carbohydrates 10-20%, and further contains crude fiber, crude ash, phosphorus, calcium, manganese, iodine, minerals and vitamins (A, D, C, E, _B2 ,
B ₆ , B ₁₂ ).

In implementing the method of the present invention, a predetermined amount of purple non-sulfur bacteria or/and purple sulfur bacteria and the vitamins mentioned above are added and mixed into the commercially available compounded feed for artificial culture of shrimp and crab as described above. All you have to do is throw the bait. The feeding method and timing are no different from feeding commercially available shrimp and crab compound feed for artificial cultivation.

According to the method of the present invention as explained above, compared to conventional artificial aquaculture methods, it is possible to significantly reduce the mortality rate during the growth process of shrimp and crabs, especially during the zoia stage, and at the same time Growth retardation or cessation can be prevented, and as a result, shrimp and crabs can be artificially cultivated in large quantities at a profitable rate.

It should be noted that the compounded feeds for artificial culture of shrimp and crabs that have been commercially available to date are not only effective in improving the survival rate during the zoia stage but also in preventing growth retardation or cessation during the growth process until adulthood. There is no bait (in fact, nothing has been developed from this point of view), and from this point of view as well, the method of the present invention can be said to be excellent.

Next, the method of the present invention will be explained using Examples and Comparative Examples.

Example 1 400 fertilized eggs of American rhinoceros shrimp were divided into four parts,
100 fish were placed in four aquariums (one was used as a control, and the rest were treated), and the water temperature in each tank was kept at 20℃, with ventilation, and 20% per day.
(volume ratio) was replaced with water and artificial culture was performed.

The food used in each tank was as follows.

Control area: Commercially available compound feed for shrimp and crab artificial culture: “Manufactured by Company A, cylindrical granules with a diameter of approximately 3 mm and a length of approximately 15 mm, composition: approximately 35% crude protein, approximately 2.0% crude fat, approximately carbohydrates. Ten
%, crude fiber approximately 5.0%, crude ash content approximately 9.0% or less, phosphorus,
Calcium, manganese, iodine, minerals, vitamins A, D, C, E, _B2 , _B6 , _B12 in trace amounts.
(hereinafter referred to as commercial feed A) to 1 g per day.
I fed it for 1.5 months.

Treatment area Feed containing 0.2 g of vitamin B ₁ , 0.2 g of para-aminobenzoic acid, 0.05 mg of nicotinic acid, and 0.01 mg of biotin per 100 g of commercially available feed A was added to 1 g per day.
I fed it for 1.5 months.

Treatment area: Rhodoseudomonas capsulitas ``Feikoken Bacteria No. 879'' per 100g of commercial feed A
1g of feed supplemented with 10g of freeze-dried bacterial cells per day.
I fed them for 1 to 1.5 months.

Treatment area: 10 of the same bacteria as above for 100g of commercial feed A
g and vitamin B ₁ 0.1 g, para-aminobenzoic acid 0.1
A feed containing 1 g of nicotinic acid, 0.05 mg of nicotinic acid, and 10 μg of biotin was fed to the rats at a rate of 1 g per day for 1 to 1.5 months.

The culture conditions in each tank were as follows.
In the control plot, molting was not completed after the third molting, growth stopped, and the chicks died at the zoia stage.

In the treated area, the molt did not complete after the 4th molt, the growth stopped, and the larva died at the zoia stage.

In the treatment area, 50% of the animals molted 10 times and reached adulthood, continuing to grow and even reaching the point where they could hold fertilized eggs.

In the treatment area, 98% of the animals molted 10 times and reached adulthood, continuing to grow and even reaching the point where they could hold fertilized eggs.

Example 2 Artificial cultivation was carried out under exactly the same conditions as in Example 1, except that the cells in the treated plot were changed to freeze-dried cells of Rhodosprirum rubrum "Kikoken Bacteria No. 878".

The culture conditions in each tank are as follows.

In the treatment area, 50% of the animals molted 10 times and reached adulthood, continuing to grow and even reaching the point where they could hold fertilized eggs.

In the treatment area, 97% of the animals molted 10 times and reached adulthood, continuing to grow and even reaching the point where they could hold fertilized eggs.

Example 3 Artificial culture was carried out under exactly the same conditions as in Example 1, except that the cells in the treated plot were changed to freeze-dried cells of Chromatium binosum "Keikoken Bacteria No. 890".

The culture conditions in each tank are as follows.

In the treatment area, 49% molted 10 times, reached adulthood, continued to grow, and even matured to the point where they could hold fertilized eggs.

In the treatment area, 88% of the animals molted 10 times and reached adulthood, continuing to grow and even reaching the point where they could hold fertilized eggs.

Example 4 The cells in the treatment area were mixed with 10 g of freeze-dried cells of Rhodoseudomonas capsulitas ``Feikoken Bacteria No. 879'' and 10 g of freeze-dried bacterial cells of Chromatium binosum ``Feikoken Bacteria No. 879'' for 100 g of commercial feed A. Artificial cultivation was carried out under exactly the same conditions as in Example 1, except that 10 g of freeze-dried bacterial cells of "Kyoyori No. 890" were used.

The culture conditions in each tank are as follows.

In the treatment area, 54% of the animals molted 10 times and reached adulthood, continuing to grow and even reaching the point where they could hold fertilized eggs.

In the treatment area, 99% of the animals molted 10 times and reached adulthood, continuing to grow and even reaching the point where they could hold fertilized eggs.

Example 5 400 spiny lobster phyllosomes (zoia stage) were divided into quarters and 100 pieces each were placed in 4 aquariums.
In the same way as above, 1 piece was placed in the control area, the rest were placed in the treated area, etc.
Artificial culture was carried out under the same conditions as in Example 1 (however, the number of days for feeding was approximately 2 months in each tank).

The culture conditions in each tank were as follows.

In the control group, 2% of the animals metamorphosed into adults with the same morphology as their parents.

In the treated area, 5% of the animals metamorphosed into adults with the same morphology as their parents.

In the treatment area, 68% of the animals metamorphosed into adults with the same morphology as their parents, and continued to grow steadily and even matured to the point where they could hold fertilized eggs.

In the treatment area, 98% of the animals metamorphosed into adults with the same morphology as their parents, and continued to grow steadily and even matured to the point where they could hold fertilized eggs.

Example 6 Artificial cultivation was carried out under exactly the same conditions as in Example 5, except that the cells in the treatment plot were changed to freeze-dried cells of Chromatium binosum "Keikoken Bacteria No. 890".

The culture conditions in each tank were as follows.

In the treatment area, 45% of the eggs metamorphosed into adults with the same morphology as their parents, and continued to grow steadily and even matured to the point where they could hold fertilized eggs.

In the treatment area, 83% of the animals metamorphosed into adults with the same morphology as their parents, and continued to grow steadily and even matured to the point where they could hold fertilized eggs.

Example 7 400 freshwater crabs that had just entered the zoia stage were divided into four, and 100 crabs each were placed in four aquariums Example 1
In the same way as above, 1 piece was placed in the control area, the rest were placed in the treated area, etc.
Artificial culture was carried out by maintaining the water temperature in each tank at 20°C, aerating the tank, and replacing the water 100% (by volume) each day.

The food used in each tank was as follows.

Control group: 1 to 5 g of the feed used in the control group of Example 1 per day.
I have been feeding to this address for 2 months.

Treatment area The feed used in the treatment area of Example 1 was fed 1 to 5 times a day.
I fed to g for 2 months.

Treatment area The feed used in the treatment area of Example 1 was fed 1 to 5 times a day.
I fed to g for 2 months.

Treatment area The feed used in the treatment area of Example 1 was fed 1 to 5 times a day.
I fed to g for 2 months.

The culture conditions in each tank were as follows.

In the control group, 5% metamorphosed and reached the megalopa stage, which resembled the parent's body shape, but the rest died during the zoia stage.

In the treated plots, 7% of the rats metamorphosed and reached the megalopa stage, which resembled their parent's body shape, but the rest died during the zoia stage.

In the treatment area, 80% metamorphose and reach the megalopa stage, where the body shape resembles that of the parent, and after that, 65% undergo further metamorphosis.
reached adulthood, and continued to grow and mature to the point where it could hold fertilized eggs.

In the treatment area, 98% metamorphose and reach the megalopa stage, where the body shape resembles that of the parent, and after that, 92% undergo further metamorphosis.
reached adulthood, and continued to grow and mature to the point where it could hold fertilized eggs.

Furthermore, even when the cells in the treated plot were changed to freeze-dried cells of Rhodospirillar rubrum "Feikoken Bacteria No. 878", the results remained almost unchanged.

In addition, the bacterial cells in the treated area were collected as chromatium.
When the cells were changed to the freeze-dried cells of Vinosum ``Kikoken Bacteria No. 890'', 78% of the cells in the treatment area reached the megalopa stage, which resembled the body shape of the parents, and 78% of the cells reached adulthood and carried fertilized eggs. Of those in the treatment area, 62% have reached maturity, 95% have reached the megalopa stage, which resembles the body shape of their parents, and 89% have reached adulthood and are capable of carrying fertilized eggs. Ta.

Example 8 Divide 400 fertilized shrimp eggs into four, put 100 eggs each into four aquariums, set one as a control group and the rest as a treatment group, same as Example 1. Conditions (however, the number of feeding days is 1 to 1.5 months for each tank, and commercially available feed A is used as a commercially available mixed feed for artificial culture of shrimp and crabs: "Manufactured by Company B, particle size approximately 2 mm, length approximately 10 mm.
mm of granules, composition - crude protein approx. 50%, crude fat approx.
5.6%, carbohydrates approximately 10%, crude fiber approximately 1.2%, crude ash approximately 10
%, phosphorus, calcium, manganese, iodine, minerals, and vitamins A, D, C, E, B ₂ , B ₆ , and B ₁₂ were changed to trace amounts of each (hereinafter referred to as commercial feed B). ) was used for artificial cultivation.

The culture conditions in each tank were as follows.

In the control plot, 5% underwent metamorphosis and molting and grew to adulthood, but the rest died at the zoia stage.

In the treated area, 8% of the animals underwent metamorphosis and molting and grew to adulthood, but the rest died at the zoia stage.

In the treatment area, 50% of the animals underwent metamorphosis and molting to reach adulthood, and continued to grow steadily and even matured to the point where they could hold fertilized eggs.

In the treatment area, 85% of the animals underwent metamorphosis and molting to reach adulthood, and continued to grow steadily and even matured to the point where they could hold fertilized eggs.

In addition, 100 g of commercially available feed B was added to the bacterial cells in the treated area.
10 g of freeze-dried bacterial cells of Rhodoseudomonas capsulitas "Feikoken Bacillus No. 879" and Chromatium vinosum "Feiken Bacillus No. 890"
When changing to 10 g of freeze-dried bacterial cells, 45% of the treated cells matured to the point of holding fertilized eggs, and 87% of the treated cells.

Example 9 Divide 400 fertilized tiger shrimp eggs into four, put 100 eggs each into four aquariums, set one as a control group and the rest as a treatment group, same as in Example 1. Conditions (However, the number of days for feeding was 1 to 1.5 months in each tank, and commercial bait A was changed to commercial bait B.)
We carried out artificial cultivation.

The culture conditions in each tank were as follows.

In the control group, 4% of the animals underwent metamorphosis and molting to reach adulthood.

In the treated area, 6% of the animals underwent metamorphosis and molting to reach adulthood.

In the treated area, 45% of the animals underwent metamorphosis and molting to reach adulthood.

In the treated area, 75% of the animals underwent metamorphosis and molting to reach adulthood.

Claims (1)

[Claims] 1. When artificially cultivating shrimp and crabs, the commercially available compounded feed used for artificially cultivating shrimp and crabs contains purple non-sulfur bacteria and/or purple sulfur bacteria and trace amounts of vitamin B ₁ , para-aminobenzoic acid, nicotinic acid,
A method for artificially cultivating shrimp and crabs, characterized in that the mortality rate of shrimps and crabs during metamorphosis and molting is reduced by feeding a feed supplemented with biotin.