JPS58175451A - Artificial raising method of lobster and crab - Google Patents

Abstract

PURPOSE:To reduce remarkably the death rate of lobsters and crabs in the growth process thereof and obtain active eggs having a high hatchability in a large amount, by adding purple nonsulfur bacteria or/and purple sulfur bacteria and vitamin B1, etc. to a mixed feed for the artificial raising. CONSTITUTION:A feed prepared by adding 0.1-10wt% purple nonsulfur bacteria or/and purple sulfur bacteria, and very small amount, e.g. several g-several tens of mg, based on 100g commercially available feed used for the artificial riasing, vitamin B1, p-aminobenzoic acid, nicotinic acid and biotin to the commercially available mixed feed for the artificial raising.

Description

[Detailed Description of the Invention] The present invention relates to an artificial culture method for shrimp and crab.
The purpose is to provide an artificial culture method that can keep the mortality rate as low as possible during the metamorphosis and molting stages of the growth process of shrimp and shrimp.Despite the fact that the harvest volume has been on the decline in recent years. Many attempts have been made to artificially farm shrimp and crabs, which are in high demand and expensive, but the reality is that no profitable artificial farming method has been established due to the high mortality rate.

The well-known shrimp, shrimp, and crabs undergo metamorphosis and molt during their growth process to grow into adults. For example, the fertilized eggs of the spiny lobster attach to the mother's abdomen and swimming limbs and are protected in the naupp. 17 (Naupliug) stage, approximately 1
At the end of the month, it becomes Phyllosoma and enters the Zoia (Zo・a) stage. During this period, the body grows larger as it repeatedly molts its skin while living a floating life, and enters the Mysia stage, during which it molts once. It becomes an adult.

Incidentally, American Caputo shrimp, Long-tailed shrimp, and Kuruma prawns also become adults through a growth process that is almost the same as that described above (However, Kuruma prawns are laid eggs in water, so they spend the narapuri period in water).

In addition, for example, the fertilized eggs of the crab attach to the abdomen and appendages of the mother's body and pass through the narapuri stage while being protected.
After about 1 to 3 weeks, the larva becomes molten and becomes independent from its mother's body, entering the zoia stage. During this stage, the larva moves its abdomen back and forth, living a floating life, and repeatedly molts its skin, increasing its size and becoming the megaloba (M@galopa) stage. During this period, they molt once and become adults.

In addition, when artificially cultivating shrimp and crabs that follow the same growth process as the above-mentioned growth process, the mortality rate during the metamorphosis and molting stages is extremely high. .

For example, in the case of artificial cultivation of Kuruma prawns, the following is true: If the fertilized eggs released from Kuruma prawns with ripe eggs are collected with a net and cultured in a culture tank, the survival rate during the Narapuri stage is The survival rate is high at about 90 chicks because they live on the egg yolk in their bodies, but in the zoia stage, the survival rate when fed with diatoms is very low at about 28 chicks, and when they reach the mysis stage, they become stronger and become animals. The survival rate when fed plankton is about 86%.

Note that the same applies to other types of shrimp and crabs, and the mortality rate during the zoia stage is extremely high. We have been conducting systematic research for many years in search of artificial aquaculture methods that can keep the mortality rate of shrimp and crabs as low as possible during the growth process, but in the process, we have When feeding by adding photosynthetic bacteria such as purple nonsulfur bacteria, purple sulfur bacteria, and trace amounts of vitamin B4, para-aminobenzoic acid, nicotinic acid, and biotinic acid to the compound feed commonly used for aquaculture. In other words, the present invention provides a method for artificially cultivating shrimp and crabs by reducing the mortality rate to an extremely low level. For this purpose, purple non-sulfur bacteria and/or purple sulfur bacteria and trace amounts of vitamin B4, para-aminobenzoic acid, nicotinic acid, and biotin are added to commercially available compounded feed used for artificial cultivation of shrimp and crabs. Shrimp characterized by feeding bait,
This is an artificial crab farming method.

Now, the structure and effects of the method of the present invention will be explained as follows. It survives in most of the five backwater locations (paddy fields, ditches, sewage, rivers, etc.) in the subtropics, and the details listed below are well known.

Purple non-sulfur bacterium - Asiorodase: Mthiorhod
aceae- has the following genera (for species belonging to each member, one representative species is listed and its mycological properties are noted ◎)01
Rhodoshade Mona x: Rhodop, aeudom
Onas genus Capgulatu: Capgulatu
S species A1 Morphological characteristics It is a short rod-shaped bacterium (width 0.5μ x length 1.0μ) that has a single flagellum and is highly motile, but depending on the type of culture medium and culture period. Long rod soft bacteria (width 0.5~0.7μ
x length 6.0 μ), which indicates a polymorphic phenomenon.

b1 Growth conditions Growth status in various media (anaerobic lighting conditions) Meat juice Ten peptone water +++ Potato medium - Ciosulfate -
Alanine + Liesin - Asparagine + Aspartic acid - Glutamic acid
life stone acid
-citric acid - geltaric acid earth acid acid decapropionic acid
＋＋＋Lactic acid
++ Succinic acid Decmalic acid Decabutyric acid Ten+ Crotonic acid Decopyruvate ++ Ethanol - Mannitol - Sorbitol
- Glucose Mannose - Fructose Decoglycerol - (Both use 0.2-concentration for the substrate) Note: ++
10 - Good growth + → Possible to grow - → Not viable C1 Physiological properties (1) Optimum growth conditions PH7.2, temperature 27℃, anaerobic lighting (10
ux] (2) Conditions for growth pH 6 [1 - 85 Temperature 23 - 39°C % Air - Anaerobic dark conditions - Lighting conditions (5) Negative Durham staining (4) Acid-fast ants (5) Production of indole (6) Generates hydrogen sulfide (7) Capable of fixing nitrogen gas (8) In nitrate medium, nitric acid is reduced to N2? 0 (9) Formation of camerase (→ liquefaction of gelatin) C (u) Hydrolysis of starch C (b) Reduced methylene blue, reduced methyl ( or benzyl) Ability to oxidize biotin pigment (b) Requires biotin, thiamine, and nicotinic acid as growth factors. So, the moving spiral shape (05
~1.5μ x length 20~10μ) 0 Ob1 growth conditions showing various phenomena Growth status in various media (anaerobic lighting conditions) Meat juice Ten peptone water Ten potato medium - Thiosulfate - Arani/ + Leusin ± Asparagine + aspartic acid ・ + glutamic acid
Namazake stone acid −
− Citric acid − Geltar acid − Vinegar
Acid Decapropionic acid
milk acid
Decasuccinic acid Decimalic acid Decbutyric acid Deccrotonic acid Decopyruvate Tenethanol + Mannitol - Sorbitol - Glucose Mannose - Fructose - Glycerol - (Both substrates are 0
・Use 2-concentration) Note: + → Viable ± → Grows υ, Did not grow) − → Unviable C1 Physiological properties (1) Optimum growth conditions pH & 8-7. O, 30-35℃ (2) Conditions for growth p'H6, 0-8, 5 aerobic to anaerobic, dark to light (3) Negative for Durham staining (4) Anti-acidic (5) Production of indole (6) Production of hydrogen sulfide (8) Nitrate media also perform denitrification, which is the opposite of nitrogen fixation by reducing nitric acid and gasifying N2↑ (9) Production of catalase (→ gelatin Liquefaction (U) Hydrolysis of starch (B) Oxidation ability of reduced methylene blue, reduced methyl (or benzyl) biolodiene dye (requires 4-biotin as a growth factor).

Purple sulfur bacteria - Shiorhoda-4: Th1orhoaace
ae- has the following genera (species includes one representative species,
0) which describes its Dutch characteristics.

1, RiOYchi x -A: Chromatium genus pinosum: vinosum species a1 Usually oval to short rod-shaped (width 1-4 μ, length 2-1.
0μ) and flagellum-induced motility. b. Growth conditions. Growth conditions in various media (anaerobic lighting conditions).
Juice - Peptone water Shima bell medium - Thiosulfate + Sodium propionate - Sodium malate + Sodium succinate
+ grape sugar
-ethanol
- (Using 0 and 2 concentrations of substrate in both cases) Note: +
→Growable - →Non-viable C1 Physiological characteristics 1) Optimal growth conditions (pH 8.2, temperature 30℃, anaerobic light (
1o, ooo lux)) 2) Conditions for growth (pH 7.6-8.8, temperature 25°C
~40°C, anaerobic lighting conditions) 5) Negative Gram stain 4) No acid-fast residue 5) No indole formation 6) Very good use of hydrogen sulfide.

7) Nitrogen fixation ability ib 8) Does not grow on nitrate medium 9) Catalase production but no gelatin liquefaction u) Starch hydrolysis not U) Oxidation of reduced methyl (or benzyl) bio-mouth paste pigment Do the following.

→No vitamin requirement. In addition, methods for harvesting purple non-sulfur bacteria and purple sulfur bacteria from hot water 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 cultivation methods are known for large-scale cultivation, and a typical example is as follows.

That is, in the nutrient source mixing tank (NH4)2s04, KH
2PO4, MgSO4-7H20, NaCl, CaCl
2.1iaHco, .

Mix and dissolve culture medium such as yeast extract to prepare a basal medium. When culturing purple sulfur bacteria, add KNa28.9H20 to this basal medium and mix with the culture medium.
111) When culturing purple sulfur-free bacteria, add acetic acid, propionic acid, lactic acid, etc. to this basic medium and transfer the culture solution to a closed, illuminated culture tank, where the purple sulfur bacteria will be cultured. Alternatively, purple non-sulfur bacteria can be transplanted and cultured.

To be more specific, the culture medium composition is (NH4)2So40.5 g, K2 for 14 tap water.
HPO40.5 g, MgSO4-7H200.2 g
, NaCl Q,s g, CaC1゜0・5 g
%MaHCOs G ・2 g1 yeast extract 0.01
In the case of purple sulfur bacteria, add 0.3 to 0.5 of Ha28.9■20 to this composition.
1) 11B・2~8 with KOH solution
・Adjust to 5 and culture ◎ If you put the above culture solution in a transparent container such as glass, seal it, and culture it for 2 days at 30℃, 5000 lux lighting, you can collect about 3 g/l of bacterial cells ◇- The general components of the bacterial cells harvested by the culture method listed in h are 55.4% protein, 2.3% fat, 55-1% sugar, 2.9% fiber, and 4.4% ash.
Adashi, the content of vitamin A is 864,600 units, U, 7
100g, vitamin C is 161 filag 〇Also, the content of vitamin B is 20orl (by the way, yeast is 1r%, chlorella is only 101%) K. In addition, carotene pigments (lycopene, rhodobin, Spirilloxanitin, etc.), and the bacterial chlorophyll content is extremely high (1000 to 500 mg9GK).

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, spraying on the target commercially available formulated feed It is convenient to add by

The amount added needs to be at least 0.1% by weight or more based on the target commercially formulated feed, and if it is less than this, it is difficult to obtain a significant effect.

The upper limit is not particularly limited, but addition of approximately 10% by weight is sufficient in practice. However, even greater effects can be obtained by adding trace amounts of vitamin B7, para-aminobenzoic acid, nicotinic acid, and biotin ginseng.

The amount of the above-mentioned vitamins added may be extremely small, and it is sufficient to add several μ9 to several 10 M per 100 g of commercially available feed.

It has not yet been theoretically elucidated 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.However, commercially available combinations It has been confirmed that adding the above vitamins to feed has almost no effect. From this point of view, the above vitamins promote the growth of purple non-sulfur bacteria and purple sulfur bacteria. It is considered to be 0th order 1
Describe the commercially available compounded feed used in the artificial cultivation of shrimp and crabs0 A commercially available compounded feed for the artificial cultivation of shrimp and crab is a cylindrical object approximately 5 tm in diameter x 15m in length with ash content,
Contains phosphorus, calcium, manganese, iodine, minerals and vitamins (Mu, D, C, L, B2, B4, B12) In carrying out the method of the present invention, the above-mentioned commercially available shrimp , purple non-sulfur bacteria or/
and adding a predetermined amount of purple sulfur bacteria and the vitamins,
Just mix and feed. The feeding method and timing are no different from feeding commercially available shrimp and crab artificial aquaculture compound feed. According to the method of the present invention as explained above, compared to the conventional artificial aquaculture method, It is possible to extremely reduce the mortality rate during the growth process of shrimp and crabs, especially during the zoia stage, and to prevent growth retardation or cessation at each stage.As a result, it is possible to produce large quantities of shrimp and crabs profitably. It is possible to cultivate them artificially.

In addition, KVi, a compounded feed for artificial culture of shrimp and crab that is currently commercially available, not only improves the survival rate during the zoia stage.
There are no feeds that are particularly effective in preventing growth retardation or cessation during the growth process until adulthood (in fact, none have been developed from this perspective), and from this point of view, the present invention The method can be said to be excellent.Next, the method of the present invention will be explained with examples and comparative examples.Example 1 400 fertilized eggs of American caputo shrimp were divided into four parts
They were placed in four aquariums (one was used as a control, and the rest were treated, sealed, and sealed), and the water temperature in each tank was maintained at 20℃, with aeration, and 20% ( Artificial culture was performed by replacing the water (volume ratio).

The food used in each tank was as follows.

Control area commercially available compound feed for artificial culture of shrimp and crab: Made by Mu Co., cylindrical granules with a diameter of approximately 5 m and a length of approximately 15,811 cm, composition: crude protein approximately 35%, crude fat 2.0%, carbohydrate approximately Excellent 10, crude fiber 5-Ols, crude ash 9-low or less, phosphorus, calcium, manganese, iodine, minerals, vitamins A, D,
Small amounts of C, 1% B2, B, B, etc. (hereinafter referred to as commercially available feed) were fed at 1 g per day for 1 to 1.5 months.

Treatment area I Commercial feed A100f [vitamin B, o.2g,
Para-aminobenzoic acid 0.1f, nicotinic acid 0.05ml
11. Bait added with biotin 0.01109 was fed to 1f per day for 1 to 1.5 months. Treatment area: Feed added with freeze-dried bacterial cells 109 was added to 1g per day for 1 to 1.5 months.
Feeding for 1.5 months and 90 treatment areas ■ 109 of the same bacterial cells as above, 0.1 g of vitamin B, 0.1 g of para-aminobenzoic acid,
A feed containing 0.osmt of nicotinic acid and 10 μf of biotin was fed at 1 g per day for 1 to 1.5 months.

The culture conditions in each aquarium were as follows: In the control plot, molting was not completed after the 3rd molt, the growth stopped, and the child died at the zoia stage; in the 90 treatment plot ■, the molting did not occur after the 4th molting. It did not complete, continued to grow, and died at the zoia stage.In treatment, 50 chicks molted 10 times and reached adulthood, and continued to grow and matured until they held 7 fertilized eggs. In the 〇 treatment, 98 chicks molted 10 times, reached adulthood, and continued to grow and mature to the point where they could hold fertilized eggs.

Example 2 Treatment area 1. Artificial culture was carried out under exactly the same conditions as in Example 1, except that the bacterial cells of Rin were changed to freeze-dried images of Rhodospirillum rubrum "Feikoken Bacteria No. 878". The culture conditions in each tank were as follows. In the same O-treated compartment, 50 sacs molted 10 times and reached adulthood, and continued to grow and mature to the point where they could hold fertilized eggs.

In treatment plot I, 97Is molted 10 times and reached adulthood, and continued to grow and mature to the point where it held fertilized eggs.

Example 5 Artificial culture was carried out under exactly the same conditions as in Example 1, except for the treatment and rotation, and the bacterial cells were changed to freeze-dried images of Chromatineum pinosum "Huikoken Bacteria No. 890". The culture conditions in each tank are as follows. In the O-treated section, 49- molted 10 times and reached adulthood, and K also matured until it continued to grow and hold fertilized eggs.

In the treatment, 88% molted 10 times and reached adulthood, and K also matured until it continued to grow and hold fertilized eggs.

Example 4 Treated area 1, 100 g of commercially available baits were added to 10 g of freeze-dried cells of Cyterodoschutes monas capsiellatus "Koukouken Bacteria No. 879" and Chromachyme pinosum. [Freeze-dried image 109 of Microtechnical Research Institute No. 890 J
Artificial culture was carried out under the same conditions as in Example 1, except for the following changes: The culture conditions in each aquarium are as follows. As they continue to grow, they mature to the point where they can hold fertilized eggs.

In the treatment, 99- molted 10 times and reached adulthood, and continued to grow and mature to the point where it could hold fertilized eggs.

Example 5 Divide 400 spiny lobster phyllocyma (zoia stage) into quarters, put 100 pieces each into 4 aquariums, and add 100 pieces in the same manner as in Example 1.
Contrast area! IID was treated as treatment zone ■, Seal, and Kaku, and artificial culture was carried out under the same conditions as in Example 1 (however, the number of feeding days was approximately 2 months in all 6 tanks).The culture conditions in each tank were as follows. ◎ In the control plot, the two buns metamorphosed into adults with the same morphology as their parents.

In treatment area ■, 5- metamorphosed into adult Kt with the same morphology as the parent.

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.

When subjected to treatment, 98% of the eggs metamorphosed into adults with the same morphology as their parents, and continued to grow steadily and mature 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 treatment and rotation and the bacterial cells in the images were changed to freeze-dried images of Chromatineum pinosum "Feikoken Bacteria No. 890"◎ The culture conditions in each tank were as follows.

In the treatment and rotation, 45 Chi metamorphosed into an adult K with the same morphology as its parents, and continued to grow steadily and even matured to the point where it could hold fertilized eggs.

In the treated plots, 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, 100 pieces each were placed in 4 aquariums, and the same procedure as in Example 1 was carried out.
Artificial culture was carried out by keeping the water temperature in each tank at 20℃, aerating, and replacing 1001% (by volume) of water per day. The feed used was as follows.

Control area The bait used in the control area of Example 1 was fed to 1-5f per day for 2 months.Treatment area I The bait used in the treatment area of Example 1 was applied to 1-59f per day for 2 months. Feed O Treatment area I The 9 baits used in the treatment area of Example 1 were fed to 1 to 59 people per day for 2 months. I fed it for 2 months.

The cultivation conditions in each tank were as follows: In the control plot, 5 chicks metamorphosed and reached the megaloba stage, which resembles the body shape of their parents, but the remaining 9 chicks died during the zoia stage.

In treatment plot I, 7% metamorphosed and reached the megaloba stage, which resembled the parent's body shape, but the rest died during the zoia stage.

In treatment plot I, 80% metamorphosed and reached the megadonkey stage, which resembled the parent's body shape, and then further metamorphosed and reached 65-year-olds, which continued to grow and mature to the point where they could hold fertilized eggs.

In the treatment area, the 98-year-olds undergo metamorphosis and reach the megadonkey stage, which resembles the body shape of their parents, and then further metamorphosis, with 92% of them reaching adulthood. , the bacterial cells of 11 treated areas were transformed into Rhodospirillum rubrum.
There was almost no change in the results even when the cells were changed to freeze-dried cells from ``Kouken Bacteria No. 878.'' In addition, the bacterial cells from 11 kg of the treatment area were replaced by Chromateam Pinosum ``Kouken Bacteria No. 878.'' No. 890
When changing to freeze-dried microorganisms, 78% of them reached the megaloba stage, which resembled the body shape of their parents in the treatment area, and 62% of them reached adulthood and even held fertilized eggs. In fact, 95% of the plants in the treatment plot reached the megaloba stage, which resembled the body shape of their parents, and 89% of the plants reached adulthood and even held fertilized eggs.

1 Example 8 Divide 400 fertilized shrimp eggs into 4 parts, put 100 eggs each into 4 aquariums, and as in Example 1, use 1 as a control group and the rest as a treated group! , bait, and seal under the same conditions as in Example 1 (however, the number of days for feeding was 1 to 1.5 months for each type, and commercially available bait A
Commercially available shrimp and crab artificial culture feed: [Manufactured by Company B]
Granular material with a particle size of about 2 mm and a length of about 10 fi, composition - crude protein sob, crude fat about 5.6%, carbohydrates about 10 t, crude fiber 1.2 t, crude ash 10%, +77 , Calcium, Manganese, Iodine, Minerals, Hitamine, Dl C, 1%
A trace amount of each of B2, B4, and 9 was changed to ``-'' (hereinafter referred to as commercial feed B). ) The culture conditions in each tank were as follows.

In the control plot, 5-thorax t=4 underwent metamorphosis and molting and grew to adulthood, but the rest died at the zoia stage.

In treatment area I, 8 chicks underwent metamorphosis and molt and grew to adulthood, but the rest died at the zoia stage.In treatment unit 90, 50 chicks underwent metamorphosis and molt and grew to adulthood, and continued to grow steadily. It has even matured to the point where it can carry fertilized eggs.

In the treatment area, 85 chinese underwent metamorphosis and molting until they reached adulthood, and after that, they continued to grow and became fully mature with fertilized eggs. Freeze-dried bacterial cells of Rhodoshademonas capsiellatus ``Hui Technol. No. 879'' (HF and lyophilized cells of Chromatimus pinosum ``Hui Technol. No. 890'') When the bacterial cells were changed to 1G1F, the number of cells that had matured to the point of holding fertilized eggs in the treated section was 45, and that of the treated section was 87-9.

Example 9 400 fertilized tiger shrimp eggs were divided into 4 parts, 100 eggs each were placed in 4 aquariums, and as in Example 1, 1 egg was used as a control group, and the rest were used as treatment areas. Artificial culture was carried out under the same conditions as in Example 1 (however, the number of feeding days was 1 to 1.5 months for each species, and commercially available feed B was changed to commercially available feed B).

The culture conditions in each tank were as follows.

In the control plot, 4 chicks underwent metamorphosis and molt and grew to adulthood.

Processing area! In this case, 6% underwent repeated metamorphosis and molting to reach adulthood, and in 90 treatment area I, 45% underwent repeated metamorphosis and molt to grow to adulthood.

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

patent applicant Foundation for Production Development Science

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 B4 and para-aminobenzoic acid. An artificial culture method for shrimp and crab, which is characterized by feeding a feed containing , nicotinic acid, and biotin.