JP2018183075A - Feed for laboratory animals for low-fluorescence and production method thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To produce a low-fluorescence experimental animal feed capable of reducing autofluorescence derived from feed and solving the problem of noise due to autofluorescence in fluorescence imaging analysis while maintaining the nutritional composition necessary for the growth of experimental animals. Provide a method. SOLUTION: In a low fluorescent experimental animal feed containing 60 to 85% by mass of a carbohydrate source, 10 to 30% by mass of a protein source and 3 to 15% by mass of fat and oil, (i) sugars 3 to 20 as the carbohydrate source. It contains 40-70% by weight of starches and (ii) one or more non-alpha starches selected from pregelatinized corn starch and rice flour, rice starch, tapioca starch, sago starch and wheat starch. It is a feed for experimental animals containing starch-containing starch and having a mass ratio of the pregelatinized corn starch to the non-pregelatinized starch in the range of 1: 2 to 10. [Selection diagram] FIG. 1A

Description

  The present invention relates to a low-fluorescence experimental animal feed and a method for producing the same.

  Conventionally, experimental animal feeds include various feeds for normal growth and reproduction, those that cause diseases such as obesity and diabetes, such as high-fat diets, and experimental animals that lack specific nutrient sources Feeds for laboratory use have been developed, and feeds for experimental animals suitable for specific uses and types of laboratory animals are used (see, for example, Patent Documents 1 and 2).

On the other hand, in various research and analysis in the fields of medicine and biology, fluorescence imaging analysis using fluorescent or luminescent labeling substances is performed, and in recent years, methods for analyzing cells and animals in a living state have been developed. ing.
However, when fluorescent imaging analysis is performed on an experimental animal in a living state (in vivo), noise due to autofluorescence derived from feed occurs in tissues other than the target site, particularly in the digestive system, which makes accurate analysis difficult. there were.
Thus, experimental animal feeds with reduced autofluorescence have been developed, but the current situation is that the degree of reduction of autofluorescence is low and accurate imaging analysis is difficult.

In addition, as a method for reducing the autofluorescence by a feed, changing the nutrient composition in a feed or fasting an experimental animal is also considered.
However, the above method cannot cope with a long-term test, and fasting should be avoided from the viewpoint of animal welfare.

  Therefore, when conducting in vivo fluorescence imaging analysis of experimental animals, feeds for low-fluorescence experimental animals that reduce feed-derived autofluorescence, enable clear fluorescence imaging analysis, and do not affect the growth of experimental animals, and Development of the manufacturing method is demanded.

JP 2003-052312 A JP 2002-335813 A

  An object of the present invention is to meet such demands, overcome the current situation, solve the above-described problems, and achieve the following objects. That is, the present invention is a low-fluorescence experimental animal feed that can reduce the autofluorescence derived from the feed while eliminating the nutritional composition necessary for the growth of the laboratory animal, and can eliminate the problem of noise due to autofluorescence in fluorescence imaging analysis. And it aims at providing the manufacturing method.

  As a result of diligent efforts to achieve the above object, the present inventors have included a specific carbohydrate source in a specific amount in a laboratory animal feed so that the growth of the experimental animal is not affected and fluorescence imaging is performed. We found that feed-derived autofluorescence in the analysis was reduced, and that it became a feed capable of clear fluorescence imaging analysis.

The present invention is based on the above findings of the present inventors, and means for solving the above problems are as follows. That is,
<1> In a low-fluorescence experimental animal feed containing a carbohydrate source of 60 to 85% by mass, a protein source of 10 to 30% by mass and an oil and fat of 3 to 15% by mass,
(I) containing 3 to 20% by weight of sugars and 40 to 70% by weight of starch as the carbohydrate source;
(Ii) The starch contains pregelatinized corn starch and at least one unpregelatinized starch selected from rice flour, rice starch, tapioca starch, sago starch and wheat starch,
The experimental animal feed is characterized in that a mass ratio of the pregelatinized corn starch and the non-pregelatinized starch is in the range of 1: 2 to 10.
<2> The experimental animal feed according to <1>, wherein the protein source is defatted casein and / or egg white powder.
<3> The experimental animal feed according to <1> or <2>, wherein the experimental animal is a rodent.
<4> The experimental animal feed according to any one of <1> to <3>, wherein the cellulose content is 5% by mass or less.
<5> The experimental animal feed according to any one of <1> to <4>, wherein the non-pregelatinized starch is rice starch or sago starch.
<6> The laboratory animal feed according to any one of <1> to <5>, further containing vitamins and minerals.
<7> In a method for producing a low-fluorescence experimental animal feed containing a carbohydrate source of 60 to 85% by mass, a protein source of 10 to 30% by mass and an oil and fat of 3 to 15% by mass,
(I) 3-20 mass% of saccharides and 40-70 mass% of starches are blended as the carbohydrate source, and (ii) pregelatinized corn starch, rice flour, rice starch, tapioca starch, sago starch as the starches And one or more non-pregelatinized starches selected from wheat starch,
A method for producing a feed for experimental animals, characterized in that a mass ratio of the pregelatinized corn starch and the non-pregelatinized starch is adjusted to a range of 1: 2 to 10.
<8> The production method according to <7>, wherein defatted casein and / or egg white powder is blended as the protein source.

  According to the present invention, it is possible to solve the above-mentioned problems in the prior art, reduce feed-derived autofluorescence while maintaining the nutritional composition necessary for the growth of experimental animals, and the problem of noise due to autofluorescence in fluorescence imaging analysis It is possible to provide a low-fluorescence experimental animal feed and a method for producing the same.

FIG. 1A is a diagram showing a result of photographing a fluorescence spectrum image at an excitation wavelength of 675 nm / an emission wavelength of 720 nm for a mouse fed with feed 2 in Test Example 4. FIG. 1B is a diagram illustrating a result of photographing a fluorescence spectrum image at an excitation wavelength of 675 nm / an emission wavelength of 720 nm for a mouse fed with feed 4 in Test Example 4. FIG. 1C is a diagram showing a result of photographing a fluorescence spectrum image at an excitation wavelength of 745 nm / an emission wavelength of 820 nm for a mouse fed with feed 2 in Test Example 4. FIG. 1D is a diagram showing a result of photographing a fluorescence spectrum image at an excitation wavelength of 745 nm / an emission wavelength of 820 nm for a mouse fed with feed 4 in Test Example 4. FIG. 2A is a diagram showing a result of photographing a fluorescence spectrum image at an excitation wavelength of 675 nm / an emission wavelength of 720 nm for a mouse fed with feed 4 in Test Example 5. FIG. 2B is a diagram showing a result of photographing a fluorescence spectrum image at an excitation wavelength of 675 nm / an emission wavelength of 720 nm for a mouse fed with feed 5 in Test Example 5. FIG. 2C is a diagram showing a result of photographing a fluorescence spectrum image at an excitation wavelength of 745 nm / an emission wavelength of 820 nm for a mouse fed with feed 4 in Test Example 5. FIG. 2D is a diagram showing a result of photographing a fluorescence spectrum image at an excitation wavelength of 745 nm / an emission wavelength of 820 nm for a mouse fed with the feed 5 in Test Example 5.

(Low fluorescent laboratory animal feed)
The low-fluorescence experimental animal feed of the present invention (hereinafter sometimes referred to as “the feed of the present invention”) contains at least a specific carbohydrate source, a protein source, and fats and oils, and if necessary, Contains ingredients.

<Carbohydrate source>
The carbohydrate source contains at least sugars and specific starches, and further contains other starches as necessary.

  The content of the carbohydrate source in the feed of the present invention is not particularly limited as long as it is 60 to 85% by mass, and can be appropriately selected. However, it is excellent in nutrition and reduces autofluorescence derived from the feed. Is preferably 65 to 80% by mass.

-Sugar-
There is no restriction | limiting in particular as said saccharides, It can select suitably, For example, sucrose, glucose, galactose, fructose, maltose, lactose etc. are mentioned. These may be used individually by 1 type and may use 2 or more types together.
As the saccharide, one prepared by a known method may be used, or a commercially available product may be used.

  The content of the saccharide in the feed of the present invention is 3 to 20% by mass, and is not particularly limited and can be appropriately selected. However, it is excellent in nutrition and can reduce autofluorescence derived from the feed. From the point which can do, 5-15 mass% is preferable.

-Starch-
The starch contains at least pregelatinized corn starch and specific non-pregelatinized starch, and further contains other starches as necessary.

  The content of the starch in the feed of the present invention is not particularly limited as long as it is 40 to 70% by mass, and can be appropriately selected, but is excellent in nutrition and reduces autofluorescence derived from the feed. From the point which can do, 50-65 mass% is preferable.

-Alpha-ized corn starch-
The pregelatinized corn starch is used for forming a feed.
The said pregelatinized corn starch may use what was prepared by the well-known method, and may use a commercial item.

  There is no restriction | limiting in particular as content in the feed of this invention of the said pregelatinized corn starch, It can select suitably, For example, 5-20 mass% etc. are mentioned.

--- Non-pregelatinized starch--
The non-pregelatinized starch is at least one selected from rice flour, rice starch, tapioca starch, sago starch and wheat starch. These may be used individually by 1 type and may use 2 or more types together.

In the present invention, the starch includes processed starch that has been processed other than pregelatinization.
The processed starch is not particularly limited and may be appropriately selected. Examples thereof include heat-treated starch, acid-treated starch, cross-linked starch, etherified starch, and esterified starch. The processed starch may be subjected to a plurality of processing treatments (crosslinking and etherification, acid treatment and crosslinking, etc.).

  Among the non-pregelatinized starches, rice flour, rice starch, and sago starch are preferable, and rice starch and sago starch are more preferable because autofluorescence can be further reduced.

  There is no restriction | limiting in particular as the kind of rice used as the raw material of the said rice flour and rice starch, For example, a sticky rice, glutinous rice, etc. are mentioned.

  As the non-pregelatinized starch, one prepared by a known method may be used, or a commercially available product may be used.

  The mass ratio of the pregelatinized corn starch and the non-pregelatinized starch is not particularly limited as long as it is in the range of 1: 2 to 10, and can be appropriately selected. The range of 1: 3-5 is preferable at the point which can reduce fluorescence.

  There is no restriction | limiting in particular as the quantity of each non-alpha-ized starch in the case of using 2 or more types of the said non-alpha-ized starch, It can select suitably.

-Other starches-
There is no restriction | limiting in particular as long as the said other starches do not impair the effect of this invention, It can select suitably, For example, pregelatinized starch other than pregelatinized corn starch, corn starch, potato starch, etc. are mentioned. These may be used individually by 1 type and may use 2 or more types together.
As the other starches, those prepared by a known method may be used, or commercially available products may be used.
There is no restriction | limiting in particular as content in the feed of this invention of the said other starches, unless the effect of this invention is impaired, It can select suitably.

<Protein source>
The protein source is not particularly limited and may be appropriately selected. Examples thereof include milk casein, defatted casein, egg white powder, and soy protein. These may be used individually by 1 type and may use 2 or more types together.
Among these, defatted casein and / or egg white powder are preferable, and egg white powder is more preferable in that autofluorescence can be further reduced.
As the protein source, one prepared by a known method may be used, or a commercially available product may be used.

  There is no restriction | limiting in particular as a method of the degreasing process of the said degreasing process casein, A well-known method can be selected suitably, For example, the degreasing process by alcohol etc. are mentioned. By the said degreasing process, content of the vitamin component contained in milk casein can be reduced. The degreased casein is sometimes referred to as vitamin-free casein.

  The content of the protein source in the feed of the present invention is not particularly limited as long as it is 10 to 30% by mass, and can be appropriately selected, but is excellent in nutrition and reduces autofluorescence derived from the feed. Is preferably 15 to 25% by mass.

<Oil and fat>
The fats and oils are not particularly limited and may be appropriately selected. Examples thereof include vegetable fats and oils such as soybean oil, corn oil, rapeseed oil, rice oil and palm oil, and animal fats and oils such as lard and beef tallow. . These may be used individually by 1 type and may use 2 or more types together.
What was prepared by the well-known method may be used for the said fats and oils, and a commercial item may be used for it.

  As content in the feed of this invention of the said fats and oils, if it is 3-15 mass%, there will be no restriction | limiting in particular, Although it can select suitably, it is excellent in nutrition and can reduce the autofluorescence derived from feed. From the point which can do, 5-10 mass% is preferable.

<Other ingredients>
Other ingredients in the feed of the present invention are not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected. For example, vitamins, minerals, cellulose, L-cystine (hereinafter, “cystine”). ), Purified feed materials such as DL-methionine and tert-butylhydroquinone, and general feed materials that are widely used. These may be used individually by 1 type and may use 2 or more types together.
As the other components, those prepared by a known method may be used, or commercially available products may be used.
The content of the other components in the feed of the present invention is not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected.

-Vitamins-
The vitamins are not particularly limited and can be appropriately selected. For example, vitamin A, vitamin D ₃ , vitamin E, vitamin K ₁ , vitamin K ₃ , vitamin B ₁ , vitamin B ₂ , vitamin B ₆ , Examples include vitamin B ₁₂ , vitamin C, biotin, folic acid, calcium pantothenate, paraaminobenzoic acid, nicotinic acid, inositol, choline bitartrate, choline chloride and the like. These may be used individually by 1 type and may use 2 or more types together.
As the vitamins, those prepared by a known method may be used, or commercially available products may be used.
As content in the feed of this invention of the said vitamins, unless the effect of this invention is impaired, there is no restriction | limiting in particular, It can select suitably.

  Specific examples of the vitamins include, for example, AIN-76 vitamin mixture (published in 1977), which is a vitamin mixture in a standard purified feed for nutritional research using mice and rats published by the National Institute of Nutrition (AIN). ), AIN-93 vitamin mixture (published in 1993) and the like.

As a typical example of the vitamin mixture, the composition of vitamins derived from the vitamin mixture in 100 g of feed supplemented with AIN-93 vitamin mixture is shown below.
・ Vitamin A ・ ・ ・ 400 (IU)
・ Vitamin D ₃ ... 100 (IU)
・ Vitamin E ・ ・ ・ 7.5 (mg)
・ Vitamin K ₁ ... 75 (μg)
・ Vitamin B ₁ ... 0.6 (mg)
・ Vitamin B ₂ ... 0.6 (mg)
・ Vitamin B ₆ ... 0.7 (mg)
・ Vitamin B ₁₂ ... 2.5 (μg)
・ Biotin 20.0 (μg)
・ Folic acid: 0.2 (mg)
・ Calcium pantothenate ... 1.6 (mg)
・ Nicotinic acid ... 3.0 (mg)
・ Choline bitartrate 0.25 (g)

-Minerals-
The minerals are not particularly limited and can be appropriately selected. For example, calcium, phosphorus, magnesium, sodium, potassium, iron, copper, zinc, manganese, molybdenum, selenium, silicon, chromium, nickel, lithium, Examples include vanadium, iodine, fluorine, boron, chlorine, sulfate radical (SO ₄ ), and sulfur (inorganic). These may be used individually by 1 type and may use 2 or more types together.
As the minerals, those prepared by a known method may be used, or commercially available products may be used.
As content in the feed of this invention of the said minerals, unless the effect of this invention is impaired, there is no restriction | limiting in particular, It can select suitably.

  Specific examples of the minerals include, for example, AIN-76 mineral mixture (announced in 1977), which is a mineral mixture in a standard purified feed for nutritional research using mice and rats published by the National Institute of Nutrition (AIN). ), AIN-93 mineral mixture (announced in 1993) and the like.

As a representative example of the mineral mixture, the composition of minerals derived from the mineral mixture in 100 g of feed supplemented with AIN-93 mineral mixture is shown below.
・ Calcium ・ ・ ・ 500 (mg)
・ Phosphorus ... 200 (mg)
・ Magnesium: 50 (mg)
・ Sodium ... 100 (mg)
・ Potassium ・ ・ ・ 360 (mg)
・ Iron: 3.5 (mg)
・ Copper ・ ・ ・ 0.6 (mg)
・ Zinc ... 3.0 (mg)
・ Manganese: 1.0 (mg)
・ Molybdenum ... 0.015 (mg)
・ Selenium: 0.015 (mg)
・ Silicon: 0.5 (mg)
・ Chromium ... 0.1 (mg)
・ Nickel: 0.05 (mg)
・ Lithium: 0.01 (mg)
・ Vanadium: 0.01 (mg)
・ Iodine: 0.02 (mg)
・ Fluorine: 0.1 (mg)
・ Boron ・ ・ ・ 0.05 (mg)
・ Chlorine: 160 (mg)
・ Sulfur (inorganic) ... 30 (mg)

-Cellulose-
As the cellulose, one prepared by a known method may be used, or a commercially available product may be used.
The content of the cellulose in the feed of the present invention is not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected, but is preferably 5% by mass or less from the viewpoint that autofluorescence can be further reduced. .

-General feed ingredients for general use-
The general feed material used in general is not particularly limited and may be appropriately selected. Examples thereof include corn flour, wheat flour, soybean meal, rice bran, bran, barley flour, fish flour, meat flour, and skim milk powder. It is done. These may be used individually by 1 type and may use 2 or more types together.
As the general feed material that is widely used, those prepared by a known method may be used, or commercially available products may be used.
The content of the general feed raw material that is widely used in the feed of the present invention is not particularly limited and can be appropriately selected as long as the effects of the present invention are not impaired.

  There is no restriction | limiting in particular as a shape, a structure, a magnitude | size, and hardness of the feed of this invention, According to an experimental animal, it can select suitably.

The feed of the present invention may be sterilized as necessary.
There is no restriction | limiting in particular as said sterilization process, A well-known method can be selected suitably, For example, the sterilization process by gamma irradiation, the sterilization process by an autoclave, etc. are mentioned.

<Target>
There is no restriction | limiting in particular as an experimental animal used as the object of the feed of this invention, For example, in addition to rodents, such as a mouse | mouth, a rat, and a hamster, a guinea pig, a rabbit, a miniature pig etc. are mentioned. However, rodents are preferred.

According to the feed of the present invention, when the fluorescence imaging analysis of the experimental animal is performed in a living state (In vivo) while maintaining the nutritional composition necessary for the growth of the experimental animal, particularly the digestive system The self-fluorescence derived from feed in can be reduced, and the problem of noise due to auto-fluorescence can be solved. Therefore, the present invention also relates to a method for reducing noise in fluorescence imaging analysis in vivo, in which an experimental animal is bred with the feed of the present invention.
Moreover, according to the feed of this invention, especially in the fluorescence imaging analysis in a wavelength range of about 600-850 nm, the influence by autofluorescence can be reduced.

  There is no restriction | limiting in particular as a manufacturing method of the feed of this invention, Although it can select suitably, It can manufacture suitably with the manufacturing method of the low fluorescence laboratory animal feed of this invention mentioned later.

(Method for producing low-fluorescence experimental animal feed)
The method for producing a low-fluorescence experimental animal feed of the present invention is a method for producing the above-described feed of the present invention (hereinafter sometimes referred to as “the feed production method of the present invention”).

The feed production method of the present invention is a method for producing a low-fluorescence experimental animal feed comprising 60 to 85% by weight of a carbohydrate source, 10 to 30% by weight of a protein source, and 3 to 15% by weight of fats and oils. ) 3-20% by weight of sugar and 40-70% by weight of starch as a carbohydrate source, and (ii) pregelatinized corn starch, rice flour, rice starch, tapioca starch, sago starch and wheat starch as starches There is no particular limitation as long as it includes using one or more kinds of non-pregelatinized starch selected and adjusting the mass ratio of the pregelatinized corn starch and the non-pregelatinized starch to a range of 1: 2 to 10. A general method can be selected as appropriate.
In addition, defatted casein and / or egg white powder may be blended as the protein source.

For example, in the case of a pellet-shaped solid feed, the raw materials are mixed and then prepared using an agitator or a conditioner so that the raw material moisture is about 10 to 25% by mass using water and steam. Next, a method of putting into an extrusion granulator such as a pellet mill or an extruder, extruding from a hole of a die, cutting the pellet having a desired length with a fixed blade or a rotary blade, and then drying is exemplified.
There is no restriction | limiting in particular as a diameter and length of a pellet, It can select suitably, For example, diameter is about 3-15 mm, length is 1 to 1.5 times or more of the said diameter, etc. are mentioned.
There is no restriction | limiting in particular as the said drying method, It can select suitably, For example, the method of drying by about 80-140 degreeC heating etc. using normal ventilation drying etc. is mentioned.

  Hereinafter, the present invention will be described with reference to test examples, but the present invention is not limited to these test examples.

(Test Example 1: Examination of raw materials-1)
Each of the following raw materials was tested for fluorescence.
<Raw material>
A: pregelatinized starch (pregelatinized tapioca starch, manufactured by Showa Sangyo Co., Ltd.)
B: Un-α-modified starch (Tapioca starch, Showa Sangyo Co., Ltd.)
C: Un-α-modified starch (Sago starch, Showa Sangyo Co., Ltd.)
D: Un-α-modified starch (Tapioca starch, manufactured by Nippon Shokuhin Kako Co., Ltd.)
E: Un-α-modified starch (Uruchi rice starch, manufactured by Joetsu Starch Co., Ltd.)
F: pregelatinized starch (pregelatinized glutinous rice starch, manufactured by Joetsu Starch Co., Ltd.)
G: Un-α-modified starch (wheat starch, manufactured by Shinshin Co., Ltd.)
H ・ ・ ・ Un-alphanated starch (potato starch)
I: Non-alpha-modified waxy corn starch (manufactured by J-Oil Mills)

<Test method>
Each raw material was charged into the microplate (the amount of each raw material was the same amount), and using IVIS (Perkin Elmer), fluorescence spectrum images at each excitation wavelength / emission wavelength shown in Table 1 were taken and compared. . In addition, the result of the comparison was evaluated by the number of “−” and “+”. Specifically, “−” indicates that no fluorescence was emitted, and the number of “+” was increased as the fluorescence emitted was higher. The results are shown in Table 1.

  From the results in Table 1, the autofluorescence of the non-α-modified starch was lower than that of the non-α-modified corn starch that is usually used as a carbohydrate source. In addition, among the non-pregelatinized starch, rice starch and sago starch had lower autofluorescence. The pregelatinized starch had higher autofluorescence than the non-pregelatinized starch.

(Test Example 2: Examination of raw materials-2)
Each of the following raw materials was tested for fluorescence.
<Raw material>
J: Milk casein (manufactured by Fonterra Limited)
K: Degreasing casein (manufactured by Biomedicals)
L ... Egg white powder (manufactured by Taiyo Chemical Co., Ltd.)

<Test method>
Each raw material was charged into the microplate (the amount of each raw material was the same amount), and using IVIS (Perkin Elmer), fluorescence spectrum images at each excitation wavelength / emission wavelength shown in Table 2 were taken and compared. . In addition, the result of the comparison was evaluated by the number of “−” and “+”. Specifically, “−” indicates that no fluorescence was emitted, and the number of “+” was increased as the fluorescence emitted was higher. The results are shown in Table 2.

  From the results in Table 2, the auto-fluorescence was lower in the defatted casein and egg white powder than in milk casein that is usually used as a protein source. In addition, the autofluorescence of the egg white powder was particularly low at an excitation wavelength of 710 nm / emission wavelength of 760 nm and an excitation wavelength of 745 nm / emission wavelength of 800 nm.

(Comparative Example 1)
As a general experimental basic feed, an improved NIH feed (produced by Oriental Yeast Co., Ltd.) was used as the feed of Comparative Example 1 (hereinafter sometimes referred to as “Feed 1”).
The composition of feed 1 is as follows.
Skim milk powder (5.0%), fish meal (10.0%), defatted soybean (10.0%), alfalfa meal (4.0%), gluten meal (3.0%), corn (24.5%) Wheat flour (32.87%), brewer's yeast (2.0%), molasses (0.75%), soybean oil (2.5%), salt (0.33%), dicalcium phosphate (1. 25%), AIN-93 mineral mix (1.05%), AIN-93 vitamin mix (1.0%).

(Comparative Example 2)
The feed in which the alfalfa meal in the feed 1 was removed was used as the feed of Comparative Example 2 (hereinafter sometimes referred to as “feed 2”).

(Comparative Example 3, Examples 1-2)
After mixing the raw materials shown in Table 3 below, water was added to 40% of the outer portion and extruded from a die hole having a hole diameter of 12 mm using a single screw extruder to obtain a solid feed. Next, the pellet-shaped solid feed of Comparative Example 3 and Examples 1-2 was obtained under normal conditions of cooling after drying with a band dryer (hereinafter, the feed of Comparative Example 3 was referred to as “Feed 3”, The feed of Example 1 may be referred to as “Feed 4” and the feed of Example 2 may be referred to as “Feed 5”).

  In Table 1, “AIN-93 Vitamin Mix” and “AIN-93 Mineral Mix” are for nutritional studies using mice and rats published by the National Institute of Nutrition (AIN) in 1993 (AIN-93). Represents a mixture of vitamins and minerals in a standard refined feed (AIN-93 refined feed).

(Test Example 3: Examination of feed-1)
The feeds 1 to 4 produced in Comparative Examples 1 to 3 and Example 1 were tested for fluorescence.
<Test method>
For feeds 1 to 4, fluorescence spectrum images at each excitation wavelength / emission wavelength shown in Table 4 were taken and compared using IVIS (Perkin Elmer). In addition, the result of the comparison was evaluated by the number of “−” and “+”. Specifically, “−” indicates that no fluorescence was emitted, and the number of “+” was increased as the fluorescence emitted was higher. The results are shown in Table 4.

  From the results of Table 4, autofluorescence occurred in the feeds of Comparative Examples 1 to 3, whereas autofluorescence was suppressed in the feed of Example 1. Therefore, it turned out that the feed of this invention can reduce the autofluorescence derived from the feed in a fluorescence imaging analysis, and can become a feed in which a clear fluorescence imaging analysis is possible.

(Test Example 4: Feed examination-2)
About the feed 2 and 4 produced in the comparative example 2 and Example 1, the autofluorescence in the mouse | mouth which fed and fed each feed was tested.
<Mouse breeding>
In a commercially available cage, feed 2 or feed 4 and water were freely ingested and bred for 1 week. In addition, wire netting slats were used for breeding to prevent contact with bedding and feces.

<Test method>
The test animals fed the test feed for 1 week were euthanized and opened. Next, using IVIS (Perkin Elmer), fluorescence spectrum images at an excitation wavelength of 675 nm / emission wavelength of 720 nm or an excitation wavelength of 745 nm / emission wavelength of 820 nm were taken and compared.

FIG. 1A (mice fed with feed 2) and FIG. 1B (mice fed with feed 4) show the results of taking fluorescence spectrum images at an excitation wavelength of 675 nm / emission wavelength of 720 nm, and FIG. FIG. 1D (mouse bred by feeding feed 4) and FIG. 1D (mouse bred by feeding feed 2) show the results of taking fluorescence spectrum images at an excitation wavelength of 745 nm / emission wavelength of 820 nm.
From the results of FIGS. 1A to 1D, the feed-derived autofluorescence in the intestine is reduced in the mouse fed with the feed 4 of Example 1 compared to the mouse fed with the feed 2 of Comparative Example 2. Therefore, by feeding and breeding the feed of the present invention, in vivo fluorescence imaging analysis of experimental animals, feed-derived autofluorescence can be reduced, enabling clear fluorescence imaging analysis. Indicated.

(Test Example 5: Examination of feed-3)
Mice were bred in the same manner as in Test Example 4 except that the feed 4 or 5 prepared in Example 1 or 2 was used as the feed, and autofluorescence was tested.

FIG. 2A (mice fed with feed 4) and FIG. 2B (mice fed with feed 5) show the results of taking fluorescence spectrum images at an excitation wavelength of 675 nm / emission wavelength of 720 nm, and FIG. FIG. 2D (mouse bred by feeding feed 5) and FIG. 2D (mouse bred by feeding feed 4) show the results of photographing fluorescence spectrum images at an excitation wavelength of 745 nm / emission wavelength of 820 nm.
From the results of FIGS. 2A to 2D, it was confirmed that the autofluorescence derived from the feed in the intestine was reduced even in the mice fed with the feed of Example 2 and raised. Therefore, also from this test example, by feeding the feed of the present invention and rearing, in vivo fluorescence imaging analysis of experimental animals, feed-derived autofluorescence can be reduced, and clear fluorescence imaging analysis is possible. It was shown to be.

(Test Example 6: Examination of feed-4)
Mice were bred with the feed of Comparative Example 1, Example 1 or 2, and the changes in the body weight of the mice over time were tested. The mice were raised in the same manner as in Test Example 4, and the body weight was measured for 34 days from the start of the breeding.
As a result, it was confirmed that the weight fluctuations of the mice fed with the feed of Comparative Example 1 and the mice fed with the feed of Example 1 or 2 were comparable. Therefore, it was confirmed that the feed of the present invention maintains the nutritional composition necessary for the growth of experimental animals.

Claims (8)

In the low-fluorescence experimental animal feed containing 60 to 85% by mass of carbohydrate source, 10 to 30% by mass of protein source and 3 to 15% by mass of fat and oil,
(I) containing 3 to 20% by weight of sugars and 40 to 70% by weight of starch as the carbohydrate source;
(Ii) The starch contains pregelatinized corn starch and at least one unpregelatinized starch selected from rice flour, rice starch, tapioca starch, sago starch and wheat starch,
A feed for laboratory animals, wherein a mass ratio of the pregelatinized corn starch to the non-pregelatinized starch is in the range of 1: 2 to 10.   The animal feed according to claim 1, wherein the protein source is defatted casein and / or egg white powder.   The experimental animal feed according to claim 1 or 2, wherein the experimental animal is a rodent.   The feed for laboratory animals according to any one of claims 1 to 3, wherein the cellulose content is 5% by mass or less.   The experimental animal feed according to any one of claims 1 to 4, wherein the non-pregelatinized starch is rice starch or sago starch.   Furthermore, the laboratory animal feed according to any one of claims 1 to 5, further comprising vitamins and minerals. In the method for producing a low-fluorescence experimental animal feed comprising a carbohydrate source of 60 to 85% by mass, a protein source of 10 to 30% by mass and an oil and fat of 3 to 15% by mass,
(I) 3-20 mass% of saccharides and 40-70 mass% of starches are blended as the carbohydrate source, and (ii) pregelatinized corn starch, rice flour, rice starch, tapioca starch, sago starch as the starches And one or more non-pregelatinized starches selected from wheat starch,
A method for producing a feed for laboratory animals, wherein a mass ratio of the pregelatinized corn starch and the non-pregelatinized starch is adjusted to a range of 1: 2 to 10.   The production method according to claim 7, wherein defatted casein and / or egg white powder is blended as the protein source.