JPH02219566A - Treated euglena and use thereof - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention relates to a Euglena treated product with improved dispersibility and a method for raising aquatic microanimals using the same. The present invention also relates to feed for aquatic micro-animals that is rich in ω-3 highly unsaturated fatty acids and various vitamins.

(Prior art and its problems) In the current aquaculture industry, microscopic underwater animals are given as feed at the same time as the embryonic eggs of fish, shrimp, and crabs are transformed into stems. In the commonly used aquaculture method, rotifers, brine shrimp, daphnia, cobepoda, etc. are fed sequentially.

However, in this aquaculture method, it is said that it is necessary to supply 50 to 100% of the weight of the larvae as feed organisms every day, and supplying this feed organism is an important task in large-scale aquaculture operations. Become.

Most commonly used in aquaculture operations, the rotifer is a zooplankton that ingests phytoplankton to proliferate. Therefore, in aquaculture facilities, a method is used to prepare food for feeding by cultivating phytoplankton, feeding it to the rotifers, and then feeding the grown rotifers to the young fish. However, cultivating large quantities of phytoplankton to feed rotifers is dependent on the amount of natural light and temperature, so it is quite difficult and unstable even if the culture tank is well managed. For this reason, it was devised to use yeast cells instead of phytoplankton (1970 Japan Society of Fisheries Science, Spring Meeting Abstracts, p. 43, No. 213), but the Shiomizu chytrid rotifer cultured using yeast cells It has been revealed that the content of ω-3 highly unsaturated fatty acids is low and there are nutritional deficiencies (Marine Science Vol. 10, No. 9, p. 740, 197
8 years). To compensate for this defect, marine yeast was developed by kneading yeast cells and oil from fish and shellfish (Japanese Patent Laid-Open No. 53-69
196) has problems such as oil oxidation and soiling of the culture tank.

However, although Euglena and various other microorganisms have excellent nutritional value, there have been major problems in handling them. That is, if this is done incorrectly, the ingredients tend to rot, and the solids tend to aggregate into solid particles or lumps. If it is too big for food, it will not be eaten, it will settle and rot, and if it is too small it will float on the water surface and will not be eaten. Furthermore, aggregates of microorganisms have conventionally been tightly bound by mucilage, which has also been one of the causes of hindering dispersibility.

(Objective of the Invention) The present invention is to provide a stable supply of a treated product of Euglena that is easy to handle.

(Elements of the Invention Although the present invention is constructed as described in the claims, Euglena treated in the present invention has not been used industrially as feed for aquatic micro-animals. However, 1)
It has an appropriate size for feeding by microscopic aquatic animals, and unlike other freshwater and seawater monofilament species, its extracellular membrane is composed of a proteinaceous membrane called vertal, making it highly digestible. 2) The amino acid composition of the constituent proteins is
It has properties similar to animal protein, containing a large amount of synthetic amino acids, and has nutritional value comparable to casein, as well as containing various vitamins and highly unsaturated fatty acids. 3) Tank culture using glucose or the like as a carbon source is possible, and growth is relatively fast.

In the present invention, Euglena is a protozoan that belongs to the genus Euglena (genus Euglena) according to the zoological classification, and includes all species, varieties, and variants belonging thereto. A typical example is Euglena gracilis.
a gracilis), x-Glenna gracilis
Bacillus gracilis (Euglena gracilis)
var, bacillaris), Euglena viridis, Astasia longa, etc. In botany, it is classified as Euglena.

The medium used for culturing such Euglena is Koren-Hallertner medium (Journal of Protozoology).
Journal of Protozoology)
Volume 14 (1967, enlarged page 17), Halatner medium (Journal of Protozoology Volume 6 (19)
Known media such as those described in 1999), p. 23) can be used. In addition, glucose, starch hydrolyzate,
Molasses, glutamic acid, acetic acid, ethanol, etc. are used, and ammonium nitrate, diammonium phosphate, diammonium phosphate, etc. are used as nitrogen sources.
Inorganic nitrogen sources such as ammonium sulfate, glutamate,
Amino acids such as aspartic acid, or organic nitrogen sources such as peptone, casamino acids, yeast extract, and corn staple liquor are appropriately combined, and calcium, magnesium,
Inorganic salts such as manganese and iron, and vitamins B1 and B12
It is also possible to use a medium containing a small amount of . It is also possible to use Euglena photosynthesis without using a carbon source.

The appropriate culture temperature for Euglena is 20 to 35°C, and the appropriate initial pH is 3.0 to 7.5. Cultivation may be carried out either under light irradiation or in the dark. Further, during culturing, it is preferable to perform shaking 50 to 250 times per minute and moderate aeration.

In the present invention, it is advantageous for later processing to concentrate the Euglena culture solution by centrifugal collection or filtration. In particular, when collecting bacteria by centrifugation, centrifugal collection under conditions of 3000Xg or less provides better dispersibility in water after freeze-drying or freezing, and single cells (20~
Since it can be dispersed up to 40 microns in diameter, it can be said to be very preferable as a feed.

Temperature is not a particular issue when it comes to freezing or freezing.

What can be said is that the effects of the present invention cannot be obtained at temperatures higher than at least 4°C, and the enzyme action seems to proceed at such temperatures, so the present invention cannot be achieved when Euglena is handled under living conditions. It seems that no effect can be obtained.

The aquatic microfauna as used in the present invention refers to freshwater or saltwater cladocerans, cyclopods, rotifers, etc., and specifically includes cladoceran daphnia and ringworm, and cyclopods such as daphnia. It refers to microscopic animals such as rotifers, rotifers, and rotifers, which are generally used as aquatic feed.

In addition, if Euglena is desired to be enriched with highly unsaturated fatty acids and various vitamins from the viewpoint of feed for aquatic micro-animals, it can be treated with fats and oils containing a large amount of such highly unsaturated fatty acids and various vitamins during cultivation. It is possible to fortify highly unsaturated fatty acids and vitamins, and as a result of their incorporation into Euglena cells, the oxidative stability of the fortifying components also improves.

(Effects of the Invention) The Euglena treated product of the present invention has excellent dispersibility in water and is therefore suitable as feed for aquatic microfauna. Furthermore, it is obvious that it has good storage stability, so this is also suitable for use as feed. Treated Euglena feed products can also be enriched with necessary nutrients.

In addition, since the dispersibility in water is improved, the treated Euglena product of the present invention has excellent processability as a nutrient apart from being used as feed, and has uses as feed and food.

(Example 1) 600 g of crucose, 210 g of diammonium hydrogen phosphate
g, magnesium sulfate heptahydrate 15g, potassium dihydrogen phosphate 15g, calcium carbonate 6g, EDTA disodium salt 1.5g, Mohr's salt 1.5g% manganese sulfate tetra-Futoku hydrate 0.54g, zinc sulfate heptahydrate Japanese product 0.75g,
Corn staple liquor 150g, vitamin B, 1
50mg and 123ooμg of Vitamin'JB were dissolved in 30L of tap water, and the solution was charged into a 50L jar fermenter (120°C, 20 minutes) and sterilized with steam. To this, Euglena gracilis (Eu
glena gracilis) was inoculated, and aerated culture was performed at pH 4, 5, and 28°C for 48 hours.

The Euglena used was Euglena gracilis NIES48, which was provided by the National Institute of Pollution and Microbial System Conservation (16-2 Onogawa, Tsukuba City, Ibaraki Prefecture), and the same Euglena can be prepared manually upon request.

After culturing, the culture solution was centrifuged to about 100% using a continuous centrifuge.
The cells were collected by centrifugation at 0xg to obtain 950 g of a semi-fluid cake with a solid content of approximately 33%.

The resulting cake was dried using a commercially available laboratory freeze dryer.
Using OOL J (manufactured by Yamato Kagaku ■, product name), -
The mixture was quickly frozen to -20°C, then freeze-dried under high vacuum conditions, and stored until use.

(Example 2) The same culture as in Example 1 was carried out to culture Euglena fortified with 2 nutrients, and the culture solution was enriched with 0% as a highly unsaturated fatty acid enrichment treatment.
．． 5% squid oil (containing 12% eicosapentaenoic acid and 1385% docosahexaenoic acid) was added to the medium 30 hours after the start of culture. As vitamin enrichment treatment, 0.1% vitamin C and 0% and 0% vitamin E were added. After the culture was completed, the bacteria were collected in the same manner as in Example 1 to obtain 965 g of bacterial cells.

The resulting cake was similarly frozen under conditions as described in Example 1 and stored until use.

Eicosapentaenoic acid (EPA) in enriched Euglena
An analysis example of the docosahexaenoic acid (DHA) content and vitamin content is shown in Table 1 in comparison with the Euglena obtained in Example 1.

Table 1: Unenriched Euglena Enriched Euglena EPA+DMA” ?, 2 28
．． 3 Vitamin C" 26.0 543.1 Vitamin E" 8.5 381.5* Percentage in total fatty acids* mg7100g (dry basis
) (Example 3) Outdoor cultivation for comparison with Euglena obtained in Example 1! ! After harvesting, dried and crushed marine chlorella was used separately to culture Shiomitsu chytrid rotifer. Culture was carried out for 8 days using a 500 L polycarbonate water pot and feeding with the freeze-dried product obtained and stored in Example 1. Feeding amount of Euglena and marine chlorella is 300,000 cells each.
/ml, 680 x IO'cells/1.

The given Euglena was easily dispersed in the water. Marine chlorella was not always well dispersed, with some floating on the water surface and others settling. Culture is done by thinning method.
The total number of rotifers is shown in Figure 1. By feeding Euglena to Shimizu rotifers, a growth rate equal to or higher than that of marine chlorella was obtained.

(Reference example, Rearing of larvae using aquatic micro-animals) A 3-week rearing test was conducted using red sea bream larvae (total length: 4.9411In+) 7 days after snoring. There were three test plots: 1) rotifer plot cultured with marine chlorella, 2) rotifer plot cultured with Euglena-treated material obtained in Example 1, and 3) rotifer plot cultured with nutrient-enriched Euglena-treated material obtained in Example 2. It was designated as a rotifer ward. A 100L polycarbonate water tank was used for the test, and 300
It housed red sea bream larvae with one tail at a time. In addition, it is known that when red sea bream are reared with the low-nutrition rotifer (cultured with baker's yeast, for example), the fish body swells around 6 m long, begins to swim frantically, and dies. Nagasaki Water Test Research Report, 2@, pp. 113-116, 197
6 years). The survival rate and changes in total length during the test period are shown in Figures 2 and 3, respectively.

As is clear from this figure, the rotifers cultured on Euglena have high nutritional value for fish, and can be said to have a significant nutritional enrichment effect.

(Example 4) A feeding test similar to that of Example 3 was conducted except that Daphnia daphnia was used instead of the rotifer.

Figure 4 shows the proliferation of Daphnia daphnia. As is clear from the figure, Euglena was a good food when fed to Daphnia daphnia.

(Example 5) A feeding test similar to that in Example 3 was conducted, except that Tigriobus was used instead of the rotifer.

Figure 5 shows the growth of Tigliobus. As is clear from the figure, Euglena was a good food even when fed to Tigriobus.

As is clear from the above examples, the Euglena treated product of the present invention is highly useful.

[Brief explanation of the drawing]

Figure 1 shows the results of culturing Shiomitsu rotifers using the Euglena-treated product of the present invention in comparison with marine chlorella, and Figures 2 and 3 show Shiomitsu rotifers cultured using the Euglena-treated product of the present invention. Figures 4 and 5 show the survival rate and total length of red sea bream larvae reared using rotifers as feed in comparison with a control plot, respectively. FIG. 3 is a diagram showing the results of culturing Daphnia daphnia or Tigriobus, respectively, and conducting a fish farming test using them as feed, in comparison with a control group. Patent applicant: Harima Chemical Industry Co., Ltd.

Claims (1)

[Scope of Claims] 1) A processed product of dispersible Euglena, which is obtained by subjecting fresh Euglena to freeze-drying or freezing treatment. 2) The treated product according to item 1 above, wherein Euglena is enriched with ω-3 polyunsaturated fatty acids or various vitamins during its culture. 3) A method for raising aquatic microanimals, which comprises feeding a processed product obtained by freeze-drying or freezing raw Euglena. 4) The method of paragraph 3, wherein the live Euglena is enriched with omega-3 polyunsaturated fatty acids or vitamins during its culture.