JPH02238855A - Color-improving feed for fish - Google Patents

Abstract

PURPOSE:To obtain a color-improving feed for fish effective in improving the color tone of fishes and crustaceans to natural color tone by adding a culture product, etc., of an astaxanthin-producing microbial strain of genus Phaffia rhodozyma and vitamins C and E. CONSTITUTION:The objective color-improving feed for fish (e.g. red sea bream, rainbow trout, coho salmon, horse mackerel or sweetfish), crustacean (e.g. Japanese shrimp), etc., can be prepared by compounding a feed with (A) one or more kinds of culture product of an astaxanthin-producing microbial strain belonging to Phaffia rhodozyma (e.g. ATCC 24202) or microbial cell, decomposed cell or disintegrated cell of the strain and (B) vitamin C and/or E in an amount at least twice the amount required from the viewpoint of the nutrition of fish.

Description

[Detailed Description of the Invention] [Industrial Application Fields] The present invention is applicable to red sea bream, rainbow trout, yellowtail, coho salmon, horse mackerel, sweetfish,
This article concerns feed for improving the color tone of shrimp (hereinafter simply referred to as fish). [Prior Art] In recent years, fish farming has been widely practiced, but the habitat and food sources are different from those of natural fish, and the fish do not exhibit the same color tone as natural fish. Therefore, methods such as adding raw red shrimp, etc. to the feed for farmed fish, or adding pigments such as canthaxanthin, cabsanthin, zeaxanthin, cabunrubin, etc. are being used. However, even if these feeds are given, a color tone that is satisfactory compared to that of naturally caught fish cannot be obtained. The red color of fish is related to astaxanthin, and it has been known for a long time that the color tone can be improved by feeding fish, shrimp, etc. that contain large amounts of this pigment. contains 70-90% water
Contains so it must be stored frozen. therefore,
When using shrimp, shrimp, etc. as raw materials for compound feed, it must be dried with a moisture content of around 10%, but during the drying process, astaxanthin contained in shrimp, shrimp, etc. changes to ascuscin, which has no coloring effect. Cheap.

[The problem that the invention aims to solve 2n] Therefore, as a result of repeated research with the aim of obtaining a feed that is easy to use and has excellent effects as a color-improving feed for fish, we have developed a culture solution and bacterial cells of astaxanthin-producing bacteria belonging to Phaffia rhodochyma. When fish were fed with feed that contained , bacterial decomposition products, and crushed bacterial cells, and in which vitamin C and vitamin E were added in an amount more than double the nutritional requirements of fish, fish species, habitat environment, and color development were observed. The present invention was completed by discovering that the color tone is similar to that of natural products regardless of the part of the body. The present invention contains one or more of a culture solution, bacterial cells, bacterial cell decomposition products, and bacterial cell fragments of astaxanthin-producing bacteria belonging to Phaffia rhodozyma, and further contains vitamin C and vitamin E.
This is a color-improving feed for fish that contains at least twice the nutritionally necessary amount of the fish. [Means for solving the problem] Phaffia rotochima (Pharr) used in the present invention
A typical astaxanthin-producing bacterium belonging to the genus Ia rhodozyma is ATCC 24202.
The American Type Cultu Collection
re Collection) Catalog of Strains 1 Sixteenth Edition 1
985 (Catalogue or Strains
I Six[eenth Edition 1985
) is a bacterium described in Many other strains can be used. To select them, inoculate yeast extract/malt extract medium, select strains that exhibit a characteristic red color, check the amount of astaxanthin accumulated by a conventional method, and use strains that have a high ability to accumulate astaxanthin. This bacterium is grown in a slightly acidic (pH 5 to 6) medium containing carbon sources such as glucose, maltose, and sucrose, yeast extract/malto extract, organic and organic nitrogen sources such as ammonium sulfate, and other trace nutrients. 15-25°C (
Astaxanthin (3,3°-dihydroxy-β. β-carotene-4,4°-dione] accumulates within the bacterial cells. The feed of the present invention uses this culture as it is or its concentrate. Alternatively, bacterial cells in a centrifuged culture solution may be used. In addition, the cell wall of bacteria can be autolyzed, treated with enzymes,
It is also possible to use astaxanthin accumulated within the bacterial cells by destroying it by either chemical treatment such as acid hydrolysis or physical treatment such as grinding, ultrasonic treatment, or pressure crushing. For self-digestion, wash the bacterial cells with water and leave the wet bacterial cells to stand in the usual manner. In addition, when enzymatic treatment is performed, enzymes capable of dissolving the cell walls of lysozyme, Bacillus circulans, etc. are brought into contact with the bacterial cells using a conventional method.9Also, acid hydrolysis treatment makes it easier to digest the bacterial cell walls within the fish body. This is done by treating with an acid such as dilute hydrochloric acid to a certain extent. Pressure crushing involves crushing the cell walls of bacteria using a commonly used pressure crusher such as a French press. Ultrasonic treatment involves treating bacterial cells with ultrasonic waves to the extent that the cell walls of the bacterial cells are destroyed. The culture solution, bacterial cells, bacterial cell decomposition products, and bacterial cell crush products of astaxanthin-producing bacteria belonging to Phaffia rhodozyma obtained as described above may be used as feed as they are, but
It is preferable to coat the astaxanthin with gelatin, beef tallow, etc. in order to prevent the astaxanthin accumulated in the bacterial cells from being oxidized. Also, before coating, antioxidants such as B
It is even better to add HT (butyl hydroxytoluene), BHA (butyl hydroxyanisole), etc. This feed is mixed with commonly used feed materials, such as fish meal, meat and bone meal, krill meal, soybean oil meal, corn gluten meal, Torula yeast, wheat flour, rice bran oil meal, vitamins, etc., and made into pellets or mash. It can be molded into mixed feed. In this case, the effective amount of astaxanthin bacterial cells of the present invention is 1% or more (preferably 1 to 30%). In addition, in order to enhance the color improvement effect of astaxanthin-producing bacteria, vitamin C and vitamin E are added in an amount that is at least twice the nutritionally necessary amount of fish. , adjust the amount of vitamin E.

The feed of the present invention is extremely useful for fish farming because, when ingested by fish, not only astaxanthin accumulated in the bacterial cells is deposited in the colored parts of the fish bodies, but also other bacterial components serve as nutritional sources for the fish. It is.

Next, an experimental example of the coloring effect of the feed of the present invention on red sea bream, rainbow trout, yellowtail, coho salmon, horse mackerel, sweetfish, and prawn will be shown.

The following experiment was conducted to examine the coloring effect of the feed of the present invention on red sea bream, rainbow trout, yellowtail, coho salmon, and horse mackerel.

(Leave below) Example 1 Effect on red sea bream epidermis The coloring effect of the feed of the present invention on red sea bream was investigated using the following method.The composition of the compound feed for red sea bream used in the experiment is shown in Table 1, and the obtained results are shown in Table 1. It is shown in Table 2.

(1) Method ■Test period May 18, 1988 to August 25 ■Test fish Red sea bream (average weight approx. 96g), 15 fish of the same weight were selected and used for each experimental section.

■Breeding conditions (breeding tank) 150ρ glass tank (breeding/breeding)
Seawater was filtered through sand and heated to 25°C using a boiler. (Aeration/Water injection) A blower was used to sufficiently aerate the water, and water was added so that the water in the tank was changed once every hour. (Baiting: Morning and evening) I threw the bait twice until the fish were satiated. (
2) Method for evaluating color development effect - Macroscopic observation of body color At the end of the color development test, the body color and color of the caudal fin were observed and the degree of color development was visually ranked.

(Ranking guide for visual coloration) +10 The body surface and caudal fin are deep red, and the abdomen is red, and the red chromaticity and color tone are equal to or higher than that of wild red sea bream.10.The body surface and caudal fin A red color can be seen, but it is not as dark as Juboshi.

The body surface and caudal fin have almost no red color and are tinged with black ■Measurement of the total amount of rotinoites on the body surface After the breeding test, 10 fish were randomly selected from each area, immediately killed, and then taken from a certain part. A certain area of epidermis (scales and epidermis 10
04/1 fish] was peeled off. The exfoliated epidermis was ground with anhydrous sodium sulfate, and the trichotinoids were extracted with acetone. ∎ The extract of chlorinoid was concentrated under reduced pressure and transferred to a separating funnel containing petroleum ether in order to transfer the carotenoid pigments into petroleum ether. The crude carotenoid-petroleum ether solution was then washed with water, dehydrated with calcium carbonate, and then concentrated under reduced pressure to obtain a certain amount of solution. The visible absorption curve of the thus obtained crude carotenoid-petroleum ether liquid was determined using a spectrophotometer.
The total amount of rotinite was determined from the absorbance value of the maximum absorption appearing near l as the specific absorption coefficient E': s-2000 (
Figure 1).

2. Coloring effect of the feed of the present invention on rainbow trout was investigated by the following method. Table 3 shows the composition of the rainbow trout feed used in the experiment, and Table 4 shows the results obtained.

(1) Method ■Test period March 25, 1988 to May 24 ■Test fish Rainbow trout (Rainbow trout) were raised to llt in January 1988 and then raised only with a compounded feed that did not contain astaxanthin.
Forty fish of the same weight were used in each experimental area, with an average weight of approximately 90 g. ■ Breeding conditions (breeding tank) 150ρ glass-lined aquarium (breeding water) pumped from a well, approximately 16°C ± 1°C
Water (aeration/water injection) A blower was used to sufficiently aerate the tank, and water was added once every hour so that the water in the tank was changed. (Feeding) Follow the Leitnitz feeding schedule twice, once in the morning and once in the evening.

(2) Coloring effect evaluation method ■ Visual observation of flesh color At the end of the coloring test, randomly select 10 fish from each section, immediately kill them, peel off the epidermis, observe the flesh color, and visually rank the degree of coloring. I did it.

(Ranking guide for macroscopic coloration) +10 Muscles are dark red, comparable to the color of wild sockeye salmon - G Red is observed in the muscles, but not as deep as ++.Almost red is observed in the muscles. Measurement of the amount of total muscle stomatinoids After the visual observation of the flesh color, a certain amount of muscle (2
g/per fish] was collected. The collected muscles were ground with anhydrous sodium sulfate, and the rough carotenoids were extracted with acetone. The crude carotenoid extract was concentrated under reduced pressure.
In order to transfer and dissolve the carotenoid pigment into petroleum ether, it was transferred to a separating port containing petroleum ether. Then, the crude carotenoid-petroleum ether solution was washed with water, dehydrated with anhydrous calcium carbonate, and then concentrated under reduced pressure to obtain a certain amount of solution. The visible absorption curve of the thus obtained crude carotenoid-petroleum ether liquid was determined using a spectrophotometer.
From the absorption value of the maximum absorption that appears near 70n, the specific absorption coefficient E':. -2000 and the total amount of oral tinoids was determined (Figure 2). 3. The coloring effect of the feed of the present invention on yellowtail was investigated using the following method. Table 5 shows the composition of the yellowtail moist pellets used in the experiment, and Table 6 shows the results obtained. (1) Method ■Test period: August 4, 1988 to January 6, 1988 ■Test fish Young yellowtail fish (average weight: approx. 150 g ), 50 fish of the same weight were used per experimental area. ■Breeding conditions (breeding tank) 3-ton concrete circular tank (breeding water)
Natural seawater filtered through sand (aeration/water injection) A blower was used to sufficiently aerate the tank, and water was added once every hour to change the water in the tank. (Baiting) Bait was cast twice, once in the morning and once in the evening, until the fish were sated.

(2) Coloring effect evaluation method ■ Visual observation of body color At the end of the coloring test, the color of the lateral line was observed, and the degree of coloring was visually ranked.

(Ranking guide for macroscopic color development) 10 Yellow in the lateral line is deep, equal to or better than natural yellowtail + Yellow is observed in the lateral line, but not as dark as 10+ Yellow is almost yellow in the lateral line. What is not allowed ■Experiment to measure the total amount of rotinoid on the body surfaceg! 41. The coloring effect of the feed of the present invention on coho salmon was investigated using the method described in Section 4 below. The composition of the moist pellets for coho salmon used in the experiment is shown in Table 7, and the obtained results are shown in Table 8.

(1) Method ■Test period January 11, 1983 to April 11 ■Test fish In December 1988, fish were tricked into sea water rather than fresh water, and then raised with moist pellets consisting of minced sardines and mixed feed. Salmon (average weight: approximately 300 g) were selected from among them, and 30 salmon were used in each experimental area. ■Breeding conditions (breeding tank) 3-ton concrete circular tank (breeding water)
Natural seawater filtered through sand (aeration/water injection) A blower was used to sufficiently aerate the tank, and water was added once every hour to change the water in the tank. [Baiting] Cast the bait twice, once in the morning and once in the evening, until the fish have eaten enough.

(2) Method for determining color development effect Judgment was made by visual observation of body color and total carotenoid content in muscle, and these measurements were conducted in accordance with Experimental Example 2.

Example 5 Addition to horse mackerel skin The coloring effect of the feed of the present invention on horse mackerel was investigated by the following method. Table 9 shows the composition of the compound feed for horse mackerel used in the experiment, and Table 10 shows the results obtained.

(1) Method ■Test period: September 6, 1988 - December 9, 1988 ■Test fish: The weight of horse mackerel (average weight approximately 40 g) that was caught in the wild in July 1988 and then raised with sardine mince. Fifty fish were used in each experimental area. ■Breeding conditions (breeding tank) 3F' capacity concrete circular tank (breeding water) Natural seawater filtered through sand (aeration/water injection) Provide sufficient ventilation with a blower, and change the water in the tank once an hour. Water was poured into the area. (Baiting) Cast bait twice, once in the morning and once in the evening, until the fish have eaten enough. (
2) Judgment method for coloring effect Judgment was made by visual observation of the epidermis and total amount of stomatinoids in the muscles. These measurements were performed according to Experimental Example 2.

(Margin below) Tsukasa hundred N3 Table lO Test results (horse mackerel) Feed category Visual judgment The amount of pigment in the epidermis + ++ ++ 30. 57. 90. 92.

Total amount of ronanoid, JJ (1/100 cJ Results) In all fish species, the color of the body surface or flesh of fish that received astaxanthin-containing Phaffia cells was redder than that of fish that were not given Phaffia cells. Total rotinoid accumulation was further increased by increasing the amount of either vitamin C or vitamin E added, but when the amount of both vitamins added at the same time was more than twice the nutritional requirement, The increase in red color and total stomatinoids accumulation was even more remarkable, indicating the enhancing effect of both vitamins on astaxanthin deposition in the fish body.

(Left below) Feed of the present invention (Faffia rhodochyma (ATCC 24202) and vitamin C and vitamin E added in an amount more than twice the nutritional requirement) was fed to failed salmon, sweetfish, and prawns. When compound feeds containing the feeds added in the proportions listed in Table 11 were prepared and fed to coho salmon, sweetfish, and prawns, an improvement in body color and flesh color was observed.

(Margin below)

Claims (1)

[Claims] (1) Culture, bacterial cells, bacterial cell decomposition products, and bacterial cell fragments of astaxanthin-producing bacteria belonging to Phaffia rhodozyma 1
Contains one or more species, and further contains vitamin C or/
and color-improving feed for fish and crustaceans (red sea bream, rainbow trout, yellowtail, coho salmon, horse mackerel, sweetfish, prawn, etc.) that contains vitamin E in an amount that is at least twice the nutritional requirement of the fish.