WO2024135672A1 - Composition for increasing sperm count and/or sperm concentration - Google Patents

Abstract

Provided is a composition for increasing sperm count and/or sperm concentration that includes a fish egg lipid preparation.

Description

Compositions for increasing sperm count and/or sperm concentration

This patent application claims priority to Japanese Patent Application No. 2022-203475, the entire contents of which are incorporated herein by reference.
The present disclosure relates to compositions for increasing sperm count and/or sperm concentration.

Reproductive health is translated as "sexual and reproductive health" and refers to a person's lifelong mental and physical condition in regards to sex and reproduction, and can also be referred to as reproductive health. The Ministry of Health, Labor and Welfare's health policy "Health Japan 21" highlights its importance as a lifelong "health promotion." In addition, the US National Institutes of Health also considers it important for men's reproductive health to address factors that may reduce fertility, not just women's, in order to protect their reproductive health and that of their partners.

　Men's reproductive health is thought to be maintained when sperm concentration, motility rate, and normal morphology rate are within the normal range. However, in modern times, there have been reports of deterioration of semen findings, such as decreased sperm count and sperm DNA fragmentation, due to various physical and mental stressors in daily life. It is known that smoking, excessive drinking, obesity, etc. affect semen findings, and that improving these lifestyle habits helps improve men's reproductive health.

The physiological activity of omega-3 polyunsaturated fatty acids such as DHA and EPA has been attracting attention in recent years, and various research results have been reported, including the effect of improving cognitive function through intake.

It has been reported that daily intake of 1,500 mg of DHA improves the progressive motility and total antioxidant capacity of sperm, and reduces the percentage of sperm with fragmented DNA (Non-Patent Document 1). It has also been reported that mice lacking an enzyme (LPAAT3) that incorporates DHA into phospholipids have abnormal sperm shape (Non-Patent Document 2). Patent Document 1 also discloses a composition that contains a large amount of phospholipids bound to DHA.

WO2021/132516

The objective of this disclosure is to provide a new means for increasing sperm count and/or sperm concentration.

The present disclosure provides a composition for increasing sperm count and/or sperm concentration, comprising a fish egg lipid preparation.

The present disclosure provides a new means for increasing sperm count and/or sperm concentration.

The study flow chart shows the process from test food allocation to analysis. The results of a semen analysis in a middle-aged group (35 to 44 years old) are shown below. The test food group showed a significant increase in the change in sperm concentration (sperm count) at 12 weeks compared to the placebo group.

In this disclosure, when a numerical value is accompanied by the term "about," it is intended to include a range of ±10% of that value. For example, "about 20" is intended to include "18-22." Numerical ranges include all values between and at the endpoints. "About" in reference to a range applies to both endpoints of the range. Thus, for example, "about 20-30" is intended to include "18-33." When percentages of ingredients are expressed in this disclosure, they are by weight unless otherwise noted.

Unless otherwise specifically specified, terms used in this disclosure have the meanings generally understood by those of ordinary skill in the art of organic chemistry, medicine, pharmacology, molecular biology, microbiology, and the like. Below are definitions of some of the terms used in this disclosure, but these definitions take precedence over the general understanding in this disclosure.

(Method of producing fish roe lipid preparation)
The fish roe lipid preparation is, for example, prepared from the roe of salmonid fish, which includes the genus Salmonella, Salmoides, Salmognathus, and Salmognathus, with Salmonella or Salmogna being more preferred. Examples of Pacific salmon include chum salmon (Oncorhynchus keta), coho salmon (Oncorhynchus kisutch), pink salmon (Oncorhynchus gorbuscha), cherry salmon (Oncorhynchus masou masou), yamame (Oncorhynchus masou masou), Taiwan salmon (Oncorhynchus masou formosanus), satsukimasu (Oncorhynchus masou ishikawae), amago (Oncorhynchus masou ishikawae), Biwa salmon (Oncorhynchus masou rhodurus), rainbow trout (Oncorhynchus mykiss), chinook salmon (Oncorhynchus tshawytscha), sockeye salmon (Oncorhynchus nerka), kokanee (Oncorhynchus nerka), and kunimasu (Oncorhynchus kawamurae). Examples of Atlantic salmon include Atlantic salmon (Salmo salar) and brown trout (Salmo trutta). In one embodiment, the fish roe lipid preparation is prepared from pink salmon (Oncorhynchus gorbuscha) roe.

In this disclosure, the term "fish roe" refers to any degree of processing, unless otherwise specified. Thus, in this disclosure, the term "fish roe lipid preparation" also includes lipid preparations made from fish roe extracts, fish roe oils, refined fish roe oils, dried products of any of these, and the like.
In a particularly preferred embodiment, the fish roe lipid preparation is prepared from salmon roe, salmon roe, or any processed product thereof, such as salmon roe extract, salmon roe extract, salmon roe oil, or refined salmon roe oil.

　The extraction of the fish roe lipid preparation from the raw material, which is fish roe or a processed product thereof, can be carried out, specifically, by mixing the raw material with an organic solvent having low polarity. Examples of the solvent having low polarity include, but are not limited to, one or more organic solvents selected from the group consisting of 60-99% aqueous ethanol, ethanol, hexane, isopropyl alcohol, ethyl acetate, acetone, ether, chloroform, and methanol. The extraction temperature with the organic solvent is 0-90°C, preferably 30-70°C. It is known that the phospholipid content in the obtained extract can be increased by manipulating the water content of the aqueous ethanol (see Oleoscience, Vol. 2, No. 2, pp. 67-74). The extraction of the fish roe lipid preparation from the raw material, which is fish roe or a processed product thereof, can also be carried out by a supercritical fluid extraction method using carbon dioxide.

As a fish roe lipid preparation, the fish roe lipid composition described in WO2021/132516 (Patent Document 1) may be used.

(Phospholipids)
In one embodiment, the phospholipid content of the fish egg lipid preparation is increased. Phospholipids refer to lipids having phosphorus in the form of phosphate esters, and include glycerophospholipids and sphingophospholipids. Representative examples of glycerophospholipids are phosphatidylcholine (PC), α-glycerophosphocholine (α-GPC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), 1-lysophosphatidylcholine (LPC-1), 2-lysophosphatidylcholine (LPC-2), and 2-lysophosphatidylethanolamine (LPE-2), and representative examples of sphingophospholipids are sphingomyelin (SM) and dihydrosphingomyelin (DHSM). The phospholipid content of the fish egg lipid preparation is, for example, about 26% or more, and may be about 30% or more, preferably about 35% or more, more preferably about 37.5% or more, and even more preferably about 40% or more. The upper limit of the phospholipid content in the fish egg lipid preparation is not particularly limited, but as the phospholipid content increases, the viscosity increases, and if the viscosity exceeds a certain level, it becomes difficult to handle during production, so the upper limit is, for example, about 50% or less.

In one embodiment, the fish egg lipid preparation contains at least one selected from the group consisting of α-GPC, SM, and DHSM, and in a preferred embodiment, contains SM and DHSM. The contents of α-GPC, SM, and DHSM are not particularly limited. The content of α-GPC in the fish egg lipid preparation is, for example, about 0.050% to 0.60%. The content of SM in the fish egg lipid preparation is, for example, about 0.8% to 2.3%. The content of DHSM in the fish egg lipid preparation is, for example, about 0.050% to 0.40%.

The phospholipids other than α-GPC, SM and DHSM contained in the fish egg lipid preparation are not particularly limited, and it is preferable that they are high in PC, PE, PI, LPC-2, etc. The PC content in the fish egg lipid preparation is, for example, about 24-45%, preferably about 26-43%, more preferably about 28-41%, and even more preferably about 30-39%. The PE content in the fish egg lipid preparation is, for example, about 0.90-2.3%. The PI content in the fish egg lipid preparation is, for example, about 0.80-1.8%. The LPC-2 content in the fish egg lipid preparation is, for example, about 0.60-3.0%.

In fish egg lipid preparations, PC is contained in a relatively large amount in phospholipids. The PC content in the phospholipids is, for example, about 74% or more, and preferably about 80% or more. In fish egg lipid preparations with an increased phospholipid content, the PC content in the phospholipids is even higher. Thus, in a preferred embodiment, the phospholipid content in the fish egg lipid preparation is about 35% or more, and more preferably about 40% or more, and the PC content in the phospholipids is about 74% or more, and preferably about 80% or more.

(Fatty Acid Composition)
The composition ratio of DHA in the constituent fatty acids of lipids of fish egg lipid preparation is relatively high. Fish eggs are known to contain DHA in the form of phospholipids or triglycerides (TG). Specifically, the composition ratio of DHA in the constituent fatty acids of lipids of fish egg lipid preparation is, for example, about 15% or more, preferably about 18% or more, more preferably about 22% or more, and even more preferably about 24% or more. The upper limit of the composition ratio of DHA in the constituent fatty acids of lipids in fish egg lipid preparation is not particularly limited, but is, for example, about 46% or less, preferably about 40% or less, more preferably about 35% or less, and even more preferably about 30% or less. In this disclosure, when the composition ratio of a specific fatty acid in the constituent fatty acids of lipids is expressed in %, it is based on the area of the chart in which the fatty acid composition is analyzed by gas chromatography, unless otherwise specified.

The composition ratio of EPA in the constituent fatty acids of the lipids in the fish egg lipid preparation is relatively low, for example, about 25% or less, preferably about 23% or less, more preferably about 21% or less, and even more preferably about 19% or less. The lower limit of the composition ratio of EPA in the constituent fatty acids of the lipids in the fish egg lipid preparation is not particularly limited, but is, for example, about 5.0% or more, may be about 6.0% or more, preferably about 8.0% or more, more preferably about 10% or more, and even more preferably about 12% or more.

Fish egg lipid preparations contain the following fatty acids: myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1, n9c), eicosenoic acid (C20:1), and docosapentaenoic acid (DPA) (C22:5). Other constituent fatty acids may include myristoleic acid (C14:1), pentadecenoic acid (C15:1), heptadecenoic acid (C17:1), docosenoic acid (C22:1), tetracosenoic acid (C24:1), linoleic acid (C18:2n-6), alpha-linolenic acid (C18:3n-3), gamma-linolenic acid (C18:3n-6), eicosadienoic acid (C20:2n-6), eicosatrienoic acid (C20:3n-6), arachidonic acid (C20:4n-6), docosadienoic acid (C22:2), etc.

Fish egg lipid preparations contain a large amount of phospholipids bound to DHA. In detail, the weight of DHA per 100 g of fish egg lipid preparation is, for example, about 10 g or more, preferably about 12 g or more, more preferably about 14 g or more, and even more preferably about 15 g or more. There is no particular upper limit to the weight of DHA per 100 g of fish egg lipid preparation, but it is, for example, about 30 g or less, preferably about 25 g or less, more preferably about 22 g or less, and even more preferably about 20 g or less.

The weight of EPA per 100 g of the fish egg lipid preparation is, for example, about 5.0 g or more, and more preferably about 6.0 g or more. There is no particular upper limit to the weight of EPA per 100 g of the fish egg lipid preparation, but it is, for example, about 20 g or less, and preferably about 15 g or less.

One of the DHA-bound phospholipids contained in fish egg lipid preparations is palmitoyl-docosahexanoyl-glycerophosphocholine (PDPC). PDPC is a phosphatidylcholine (38:6) in which one of the acyl groups at C-1 and C-2 is hexadecanoyl (16:0) and the other is docosahexanoyl (22:6).

The other DHA-bound phospholipid contained in fish egg lipid preparations is stearoyl-docosahexanoyl-glycerophosphocholine (SDPC). SDPC is a phosphatidylcholine (40:6) in which one of the acyl groups at C-1 and C-2 is stearoyl (18:0) and the other is docosahexanoyl (22:6).

Another DHA-bound phospholipid contained in fish egg lipid preparations is oleoyl-docosahexanoyl-glycerophosphocholine. Oleoyl-docosahexanoyl-glycerophosphocholine is a phosphatidylcholine (40:7) in which one of the acyl groups at C-1 and C-2 is oleoyl (18:1) and the other is docosahexanoyl (22:6).

Another DHA-bound phospholipid contained in fish egg lipid preparations is didocosahexanoyl-glycerophosphocholine. Didocosahexanoyl-glycerophosphocholine is a phosphatidylcholine (44:12) in which both the C-1 and C-2 acyl groups are docosahexanoyl (22:6).

One of the phospholipids contained in fish egg lipid preparations is an ether-type phospholipid. An ether-type phospholipid is a phospholipid that has a hydrocarbon chain formed by an ether bond. In particular, a glycerophospholipid that has a hydrocarbon chain formed by a vinyl ether bond at the sn-1 position and a fatty acid bonded to the sn-2 position is called a plasmalogen.

In one embodiment, the fish egg lipid preparation contains one selected from the group consisting of DHA-bound ether-type phospholipids and EPA-bound ether-type phospholipids.

(Ingredients other than lipids)
The fish egg lipid preparation may contain components other than lipids. The components other than lipids are proteins and inorganic substances such as sodium, potassium, and phosphorus. The lipid refers to a substance of biological origin that is soluble in a non-polar solvent, and includes simple lipids, complex lipids, and derived lipids (fatty acids, terpenoids, steroids, carotenoids, etc.).
The content of components other than lipids in the fish egg lipid preparation is, for example, 10.0% or less, preferably 8.0% or less, more preferably 7.5% or less, and even more preferably 7.0% or less.

The fish egg lipid preparation may contain astaxanthin. The content in 100 g of the fish egg lipid preparation is, for example, about 0.7 mg or more, preferably about 1.0 mg or more, more preferably about 1.2 mg or more, and even more preferably about 1.5 mg or more. There is no particular upper limit to the content of astaxanthin in 100 g of the fish egg lipid preparation, but it is, for example, about 23 mg or less, preferably about 20 mg or less, more preferably about 10 mg or less, and even more preferably about 5.0 mg or less.

(Application)
As shown in the examples below, the ingestion of the composition of the present disclosure can increase the sperm count and/or sperm concentration in the semen of a subject. Therefore, the composition of the present disclosure can be used to suppress the decrease in sperm count and/or sperm concentration and maintain an appropriate sperm count and/or sperm concentration in a subject who is in a condition that reduces the sperm count and/or sperm concentration, such as aging, fatigue, and disease.

The sperm count and/or sperm concentration may be measured by semen analysis, for example, by referring to "Human Semen Examination and Procedures, WHO Laboratory Manual, 5th Edition." The sperm count and/or sperm concentration of the subject may be normal or below normal. For example, the sperm concentration of the subject may be 50 million/ml. The manual may be consulted for normal values of sperm count and/or sperm concentration. For example, the lower limit of the sperm count standard may be 39 million/ejaculate, and the lower limit of the sperm concentration standard may be 15 million/ml.

The subject is typically a human male. The subject's age is not limited, but may typically be 20 years or older, 25 years or older, 30 years or older, or 35 years or older, and may be 65 years or younger, 60 years or younger, 55 years or younger, 50 years or younger, 45 years or younger, or 44 years or younger. In some embodiments, the subject is a male 44 years or younger, e.g., a male 25 years or older and 44 years or older, or a male 35 years or older and 44 years or younger. In some embodiments, the subject is a male in middle age. The subject may or may not have been diagnosed with male infertility. The subject may or may not have been undergoing infertility treatment.

The composition of the present disclosure may be a pharmaceutical composition. The method of administration of the pharmaceutical composition is not particularly limited, and is preferably oral administration, transdermal administration, or nasal administration, and more preferably oral administration. Dosage forms for oral administration include granules, fine granules, powders, coated tablets, tablets, powders, soft capsules, hard capsules, microcapsules, chewable tablets, liquids, suspensions, emulsions, etc. Dosage forms for transdermal administration include patches, tapes, sprays, lotions, creams, ointments, liquids, emulsions, suspensions, etc. Dosage forms for nasal administration include nasal drops, nasal sprays, etc.

These dosage forms are manufactured by formulating the formulation in the usual manner. In addition, various pharma- ceutically acceptable formulation substances can be blended according to the formulation needs. The formulation substances can be appropriately selected depending on the formulation, and examples include buffering agents, surfactants, stabilizers, preservatives, excipients, diluents, additives, disintegrants, binders, coating agents, lubricants, flavoring agents, sweeteners, solubilizers, etc.

The composition of the present disclosure may also be a food composition. The food composition may be in the form of a typical processed food. For example, it may be a solid food, or a food to be consumed as a liquid such as a beverage, a drinkable supplement, a powdered beverage, or a soup. Specifically, it may be consumed as, for example, juice, confectionery, jelly, tablet, dressing, seasoning, etc.

Such foods may also be provided as health functional foods or dietary supplements. Health functional foods include, for example, foods for specified health uses, foods with nutrient functions, and foods with functional claims. Health functional foods may be labeled as being used for purposes such as supporting the maintenance of sperm count and/or sperm concentration, and maintaining sperm count and/or sperm concentration that decrease with age. The labeling may be directly on the packaging, container, label, tag, or insert accompanying the product, or indirectly through advertising and promotional activities. Dietary supplements include, for example, nutritional supplements and health supplements.

The amount of intake of the composition of the present disclosure can be about 100 mg to 10,000 mg of fish egg lipid preparation per day, preferably about 300 mg to 5,000 mg, more preferably about 500 mg to 2,500 mg, even more preferably about 700 mg to 1,500 mg, for example about 1,000 mg.

Alternatively, the amount of the composition of the present disclosure taken per day can be about 20 mg or more of DHA, preferably about 50 mg or more, more preferably about 100 mg or more, and even more preferably about 150 mg or more. The amount of the composition of the present disclosure taken per day can be about 2000 mg or less of DHA, preferably about 1000 mg or less, more preferably about 750 mg or less, even more preferably about 500 mg or less, even more preferably about 300 mg or less, and especially preferably about 200 mg or less. In one embodiment, the amount of the composition of the present disclosure taken per day is about 150 mg (e.g., 135 mg to 165 mg) or about 160 mg (e.g., 144 mg to 176 mg) of DHA.

Furthermore, the intake amount of the composition of the present disclosure can be an amount that allows about 10 mg or more of EPA to be ingested per day, preferably about 30 mg or more, more preferably about 50 mg or more, and even more preferably about 60 mg or more. Further, the intake amount of the composition of the present disclosure can be an amount that allows about 500 mg or less of EPA to be ingested per day, preferably about 300 mg or less, and more preferably about 150 mg or less. In one embodiment, the intake amount of the composition of the present disclosure is an amount that allows about 60 mg (e.g., 54 mg to 66 mg) or about 100 mg (e.g., 90 mg to 110 mg) of EPA to be ingested per day.

The daily intake of each component may be contained in one composition, or may be dispersed across multiple compositions ingested in one day. For example, the daily intake of each component of the composition may be dispersed across 2 to 15, preferably 3 to 10, e.g., 4 capsules, etc., ingested in one day.

The composition of the present disclosure may be taken once or multiple times. When taken multiple times, for example, it may be taken once to several times a day, for example, once, twice or three times a day, daily or every few days, for example, every 1, 2, 3 or 7 days. The duration of intake is not limited, and for example, it may be taken continuously for a period during which an increase in sperm count or sperm concentration is required, for example, for one week or more, preferably one month or more, more preferably two months or more, and particularly preferably three months or more. There may be periods during which intake is discontinued. In one embodiment, the composition of the present disclosure is taken daily for at least 12 weeks.

The compositions of the present disclosure can be used alone or in combination with one or more additional ingredients. "Combining" ingredients means not only the use of a dosage form containing all of the ingredients and the use of a combination of dosage forms containing each ingredient separately, but also the simultaneous intake of each ingredient or delayed intake of any ingredient, so long as they are used for the same purpose. It is also possible to combine two or more additional ingredients. For example, a composition containing one or more additional ingredients in addition to a fish roe fat preparation may be used.

The composition of the present disclosure may further contain astaxanthin. The intake amount of astaxanthin may be about 50 μg or more, preferably about 100 μg or more, more preferably about 200 μg or more per day, and may be about 1000 μg or less, preferably about 400 μg or less, for example, about 200 μg (for example, 180 μg to 220 μg). Astaxanthin may be contained in the fish egg lipid composition, but astaxanthin from another source may also be used. For example, astaxanthin extracted from natural products such as krill, salmon, trout, Adonis rosa, red yeast, and Haematococcus algae, or a synthetic product may be used, and preferably, astaxanthin is contained in the pigment of Haematococcus algae. The extraction solvent for obtaining astaxanthin from natural products may be an aqueous solvent or an organic solvent. Examples of organic solvents that can be used include methanol, ethanol, isopropanol, acetone, 1,3-butylene glycol, ethylene glycol, propylene glycol, glycerin, ethyl acetate, ether, and hexane. Supercritical carbon dioxide can also be used. These solvents can be used alone or in combination of two or more.

Specific examples of Haematococcus algae include Haematococcus pluvialis, Haematococcus lacustris, Haematococcus capensis, Haematococcus droebakensis, and Haematococcus zimbabwiensis. Commercially available Haematococcus algae extracts can also be used, such as ASTOTS-S, -5O, -10O, etc. manufactured by Fuji Film Co., Ltd., Astareal Oil 50F, 5F, etc. manufactured by Fuji Chemical Industry Co., Ltd., Astaxanthin-5C, 20C, etc. manufactured by Oryza Oil & Fat Chemical Co., Ltd., Astaxanthin 5% Oil manufactured by Bioactives Japan, Astabio ^{(registered trademark)} AR1, AR5, etc. manufactured by Biogenic Co., Ltd. Also, krill-derived extracts include Astax-S manufactured by Marine Daio Co., Ltd.

In one aspect, a composition for increasing sperm count and/or sperm concentration is provided comprising a fish egg lipid preparation.
In one aspect, there is provided a method for increasing sperm count and/or sperm concentration comprising ingesting a fish egg lipid preparation to a subject in need thereof.
In one aspect, a fish egg lipid preparation for increasing sperm count and/or sperm concentration is provided.
In one aspect, there is provided a use of a fish egg lipid preparation for increasing sperm count and/or sperm concentration.
In an aspect there is provided a use of a fish egg lipid preparation in the manufacture of a composition for increasing sperm count and/or sperm concentration.

In one embodiment, a composition for increasing sperm count and/or sperm concentration containing DHA is provided, which is prepared so that the daily intake amount is about 100 mg to 750 mg of DHA. Other ingredients that may be included in the composition and the method of using the composition are the same as described above.

In one aspect, a method is provided for increasing sperm count and/or sperm concentration comprising administering to a subject in need thereof a daily intake of about 100 mg to 750 mg of DHA.
In one embodiment, a DHA for increasing sperm count and/or sperm concentration is provided, characterized in that the daily intake is about 100 mg to 750 mg of DHA.
In one embodiment, there is provided a use of DHA for increasing sperm count and/or sperm concentration, characterized in that the daily intake is between about 100 mg and 750 mg of DHA.
In one aspect there is provided the use of DHA in the manufacture of a composition for increasing sperm count and/or sperm concentration, characterized in that the composition provides a daily intake of about 100 mg to 750 mg of DHA.

For example, the following embodiment is provided.
[1] A composition for increasing sperm count and/or sperm concentration, comprising a fish egg lipid preparation.
[2] The composition according to paragraph 1, which is prepared so that the daily intake amount is about 100 mg to 750 mg (preferably about 150 mg to 750 mg, more preferably about 150 mg to 200 mg) of DHA.
[3] A composition for increasing sperm count and/or sperm concentration, comprising DHA, the composition being prepared so that the daily intake amount is about 100 mg to 750 mg of DHA.
[4] The composition according to any one of claims 1 to 3, which is prepared so that the daily intake amount is about 150 mg (or 135 mg to 165 mg) of DHA.
[5] The composition according to any one of claims 1 to 3, which is prepared so that the daily intake amount is about 160 mg (or 144 mg to 176 mg) of DHA.
[6] The composition according to any one of items 1 to 5, which is formulated to provide a daily intake of about 30 mg to 300 mg (preferably about 60 mg to 300 mg, more preferably about 60 mg to 150 mg) of eicosapentaenoic acid (EPA).
[7] The composition according to any one of items 1 to 6, which is formulated to provide a daily intake of about 60 mg (or 54 mg to 66 mg) of EPA.
[8] The composition according to any one of items 1 to 6, which is formulated to provide a daily intake of about 100 mg (or 90 mg to 110 mg) of EPA.
[9] The composition according to any one of items 1 to 8, which is formulated to provide a daily intake of about 100 mg to 750 mg (preferably about 150 mg to 750 mg, more preferably about 150 mg to 200 mg) of DHA and about 30 mg to 300 mg (preferably about 60 mg to 300 mg, more preferably about 60 mg to 150 mg) of EPA.
[10] The composition according to any one of items 1 to 9, which is formulated to provide a daily intake of about 100 mg to 750 mg of DHA and about 30 mg to 300 mg of EPA.
[11] The composition according to any one of items 1 to 10, which is formulated to provide a daily intake of about 150 mg (or 135 mg to 165 mg) of DHA and about 60 mg (or 54 mg to 66 mg) of EPA.
[12] The composition according to any one of items 1 to 10, which is formulated to provide a daily intake of about 160 mg (or 144 mg to 176 mg) of DHA and about 100 mg (or 90 mg to 110 mg) of EPA.
[13] The composition according to any one of items 1 to 12, further comprising astaxanthin.
[14] The composition described in item 13, wherein the astaxanthin is derived from a pigment of Haematococcus algae.
[15] The composition according to item 13 or 14, which is prepared so that the daily intake amount of astaxanthin is about 100 μg to 1000 μg (preferably about 200 μg to 1000 μg, more preferably about 200 μg to 400 μg).
[16] The composition according to any one of items 13 to 15, which is prepared so that the daily intake amount of astaxanthin is about 100 μg to 1000 μg.
[17] The composition according to any one of items 13 to 16, which is prepared so that about 200 μg (or 180 μg to 220 μg) of astaxanthin is ingested per day.
[18] The composition according to any one of items 1 to 17, comprising a fish egg lipid preparation containing about 30% or more (preferably about 40% or more) of phospholipids.
[19] The composition according to any one of items 1 to 18, wherein the fish egg lipid preparation contains DHA-bound phospholipid and the composition ratio of DHA to the constituent fatty acids of the lipid is about 15% or more.
[20] The composition according to any one of items 1 to 19, wherein the fish egg lipid preparation comprises at least one selected from the group consisting of α-GPC, SM, and DHSM (preferably the group consisting of SM and DHSM).
[21] The composition described in any one of items 18 to 20, wherein the phospholipid contains about 74% or more PC.
[22] The composition according to any one of items 1 to 21, wherein the DHA composition ratio of the constituent fatty acids of lipids in the fish egg lipid preparation is about 15% or more, and the EPA composition ratio is about 5 to 25%.
[23] The composition according to any one of items 1 to 22, which is taken for at least 12 weeks.
[24] The composition according to any one of items 1 to 23, which is taken daily.
[25] The composition described in any one of items 1 to 24, which is to be ingested by a healthy male.
[26] The composition according to any one of items 1 to 25, which is to be ingested by a middle-aged male.
[27] The composition according to any one of items 1 to 26, which is to be taken by a male aged 35 to 44.
[28] The composition according to any one of items 1 to 27, which is a pharmaceutical composition.
[29] The composition according to any one of items 1 to 27, which is a food composition.
[30] A method for increasing sperm count and/or sperm concentration, comprising administering a fish egg lipid preparation to a subject in need of such an increase.
[31] A fish egg lipid preparation for increasing sperm count and/or sperm concentration.
[32] Use of a fish egg lipid preparation for increasing sperm count and/or sperm concentration.
[33] Use of a fish egg lipid preparation in the manufacture of a composition for increasing sperm count and/or sperm concentration.
[34] A composition for increasing sperm count and/or sperm concentration, comprising DHA, the composition being prepared so that about 100 mg to 750 mg of DHA is ingested per day.
[35] A method for increasing sperm count and/or sperm concentration, comprising having a subject in need of such an increase ingest about 100 mg to 750 mg of DHA per day.
[36] DHA for increasing sperm count and/or sperm concentration, characterized in that a daily intake amount of about 100 mg to 750 mg of DHA is administered.
[37] Use of DHA for increasing sperm count and/or sperm concentration, characterized in that a daily intake amount of about 100 mg to 750 mg of DHA is administered.
[38] Use of DHA in the manufacture of a composition for increasing sperm count and/or sperm concentration, comprising a daily intake of about 100 mg to 750 mg of DHA.

All documents cited herein are hereby incorporated by reference.
The present invention will be described in more detail below with reference to the following examples, but the present invention is not limited to these examples. The above descriptions are all non-limiting, and the present invention is defined in the appended claims, and various modifications are possible without departing from the technical spirit of the present invention.

<Production of fish roe lipid preparation from salmon roe>
Aqueous ethanol (150 L) was added to frozen and crushed raw pink salmon (Oncorhynchus gorbuscha) roe (15 kg) for extraction, and the residue was filtered off. The residue was returned to the extractor, and ethanol (75 L) was added again for extraction. The solvent was removed from the filtrate under reduced pressure to produce a composition (40 PL) (Table 1) containing approximately 40% phospholipids in the red oily substance.

*Quantitative values are the total amount of lipids containing each fatty acid and free fatty acids.

<Production of soft capsules>
Haematococcus algae pigment (astaxanthin) was added to the 40PL to produce a soft capsule "ONO-SR/AST" containing the ingredients shown in Table 2. The daily intake of salmon roe oil was set at 1000 mg. In addition, a placebo capsule containing safflower oil instead of 40PL and astaxanthin was produced.

The amounts of phospholipid classes contained in ONO-SR/AST are shown in Table 3.

The specifications of ONO-SR/AST are as shown in the table below.

<Clinical Trials>
Subjects and Methods 1. Subjects The subjects were 110 healthy men aged 35 to 65 years. There were 55 subjects in each group. Regarding semen testing, the target was 30 subjects in each group, taking into consideration the need to obtain consent for the test. Prior to the study, a consent document was provided to the subjects, the purpose and content of the study were fully explained, and written consent was obtained from the subjects of their own free will. Based on the results of the pre-tests of subjects who had given consent, subjects who did not fall under the following exclusion criteria were selected.
(1) People who regularly consume health foods rich in DHA, EPA, and astaxanthin.
(2) Anyone who is at risk of developing an allergic reaction to the research.
(3) Persons taking medications that may affect the results of the study (such as lipid-lowering drugs, antipsychotics, antianxiety drugs, antidepressants/manic drugs, antiparkinsonian drugs, antiepileptic drugs, anticoagulants, etc.).
(4) Persons with a history of serious illness that required medical treatment (e.g. malignant tumors, diabetes, liver disease (hepatitis), kidney disease, heart disease, etc.).
(5) Anyone with a history or current medical history of drug or alcohol dependence.
(6) Those who are deemed unsuitable as subjects based on the clinical test values and measurements from the preliminary examination.
(7) Those who have donated or transfused 200 mL or 400 mL of whole blood within three months prior to the pre-examination.
(8) Individuals who have participated in other clinical research within one month of obtaining consent to participate in this study, or who plan to participate in other clinical research after obtaining consent to participate in this study.
(9) Individuals who are deemed unsuitable as subjects based on their responses to a lifestyle questionnaire.
(10) Subjects who are unable to adhere to contraception during the study period.
(11) Any other person who the responsible physician deems unsuitable as a subject.

2. Study Foods The study foods were of two types: test food (ONO-SR/AST) and placebo. The test food contained 1000 mg of salmon roe oil (containing 150 mg of DHA and 60 mg of EPA) and 200 μg of astaxanthin as an additional ingredient in the daily intake amount (1 bag, 4 tablets).

3. Study design This study was a randomized, double-blind, placebo-controlled, parallel-group comparative study with an intake period of 12 weeks. The allocation manager, who was not directly involved in the study, used a random number table to create an allocation table for each of the groups of subjects who underwent semen testing (60 subjects) and those who did not undergo semen testing (50 subjects), and assigned allocation numbers to the study foods based on this table. The allocation table was sealed by the allocation manager and kept sealed until the subjects and data were fixed. Blindness was maintained for the principal investigator, subjects, medical facility staff, and all other staff involved in the study.

Before the intervention began, all subjects in both groups underwent pre-examinations, including physical measurements (height, weight, BMI calculation), physical examinations (blood pressure, pulse rate), various clinical tests (hematology, blood biochemistry, urine tests), and questionnaires (SF-36, POMS2-AS). Those who underwent semen testing also underwent semen testing (semen volume, reaction (pH), sperm concentration, motility, rate of abnormal sperm, stained sperm specimens and photography), and the values obtained were used as the baseline.

Each subject was randomly assigned to take one bag of the study food (four tablets) per day after meals with water or lukewarm water, without chewing, throughout the 12-week intervention period. During the intervention period, subjects were asked to keep a diary recording their intake of the study food, changes in their physical condition, and medication use. Six and 12 weeks after the start of intake, all subjects visited the hospital for physical measurements, physical examinations, questionnaires, and semen tests. After 12 weeks, in addition to the above, clinical tests, special tests, SF-36, and a post-ingestion questionnaire (questionnaire on impressions after ingesting the study food) were also conducted.

4. Evaluation of efficacy The efficacy of the study food was evaluated using semen analysis (sperm concentration, motility, rate of abnormal sperm) as the primary endpoint, and SF-36, POMS2-AS, a post-ingestion questionnaire, luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone measurements as secondary endpoints.

SF-36 was used to assess health-related quality of life. SF-36 asks questions about health and generates eight subscales (physical functioning (PF), role functioning (physical) (RP), bodily pain (BP), general health (GH), vitality (VT), social functioning (SF), role functioning (mental) (RE), and mental health (MH)). The summary score showing the physical aspect of QOL (physical summary score: PCS), the summary score showing the mental aspect of QOL (mental summary score: MCS), and the summary score showing the role/social aspect of QOL (role/social summary score: RCS) were calculated using a scoring method (Web Scoring System) based on the 2017 national standard values. A higher score indicates a higher QOL.

The short version of the POMS2 for adults scores the mood state over the past week on seven scales (anger-hostility (AH), confusion-bewilderment (CB), depression-depression (DD), fatigue-lethargy (FI), tension-anxiety (TA), vitality-vigor (VA), friendliness (F)), and standardized scores (T-scores) and TMD scores were calculated according to the POMS2 Japanese manual.

In the post-ingestion questionnaire, participants were asked to answer each question about "body movement/changes," "sensation/pain/concentration," and "sleep/eyes/other" on a four-point scale: "0: not felt," "1: felt a little," "2: felt," and "3: felt a lot."

The subjects for efficacy analysis were those who completed all of the specified study schedules and contents, excluding subjects who met the following analysis exclusion criteria.
(1) Those whose intake rate of the study food was less than 80%.
(2) Those who have engaged in behavior that significantly undermines the reliability of the test results, such as missing diary records.
(3) Those who were found to meet the exclusion criteria after inclusion in the study, or those who were found to be unable to comply with the restrictions during the study period.
(4) Any other person for whom there are clear reasons that make it appropriate to exclude them.

5. Safety Evaluation Based on the interviews, physical measurements, and physical examination results on each visit, as well as the entries in the diary, the responsible physician treated all undesirable or unintended injuries, illnesses, or symptoms thereof that occurred in the subjects during the intake period as adverse events. In addition, clinical tests were performed for the following items: Hematological tests (white blood cell count, red blood cell count, hemoglobin, hematocrit, platelet count), blood biochemistry tests (total protein, albumin, total bilirubin, direct bilirubin, indirect bilirubin, ALP, AST (GOT), ALT (GPT), LD, γ-GT, total cholesterol, triglycerides, HDL-cholesterol, LDL-cholesterol, urea nitrogen, creatinine, uric acid, Na, K, CL, blood glucose, HbA1c (preliminary test only)), and urine test (protein (qualitative), sugar (qualitative), occult blood reaction (qualitative)). The judgment of whether the test values of each subject were abnormal fluctuations corresponding to adverse events was made by the responsible physician based on the standard values set by the medical institution, with reference to the criteria for judging abnormal fluctuations set by the Japanese Society of Chemotherapy, CTCAE v5.0-JCOG, and the judgment classification of the Japanese Society of Ningen Dock (revised on April 1, 2018, partially revised on December 14, 2018). In addition, the responsible physician investigated and considered the relevance to the study food, and judged whether each adverse event was a side effect of the study food. For adverse events, subjects who had consumed the study food at least once were included in the safety analysis. For measurement and test values, subjects who had completed all of the specified study schedules and contents were included.

6. Statistical analysis: Even if the test values and measurements were considered to be outliers, they were used in the evaluation. If the cause was clearly a measurement error, that time point was excluded from the analysis. If there were missing values, they were not imputed and were not included in the analysis as missing values.

　A chi-square test was used to compare gender at baseline between groups. For other items, a two-sample t-test was used to compare the change from baseline at each time point after intake between groups. For the post-intake questionnaire, the actual values measured at week 12 were compared between groups using the Mann-Whitney U test. For reference, the change at week 12 when the baseline was set at 0 was evaluated using the Wilcoxon signed rank sum test.

In order to examine the effect on sperm depending on age, a subgroup analysis was conducted based on age at the time of the pre-test. The Ministry of Health, Labour and Welfare's health policy "Health Japan 21" classifies subjects aged 25-44 as prime age and 45-65 as middle age, and accordingly, a subgroup analysis was conducted by dividing the subjects into prime age (35 to 44 years old) and middle age (45 to 65 years old). In addition, an analysis of post-intake questionnaires was conducted for those aged 40 years or older. No adjustment for multiplicity was made in each analysis. The statistical analysis software used was Microsoft Excel 365 (Microsoft, USA) and IBM SPSS statistics 26 for windows (IBM). The statistical significance level was set at 5% on both sides.

Results 1. Subjects Figure 1 shows the study flow chart, covering the process from subject selection and test food allocation to analysis. Preliminary tests were conducted on 204 subjects living in the Kanto region who provided written consent to participate in the study. From these results, 110 subjects (55 in each of the test food group and placebo group) who met the inclusion criteria and did not violate any of the exclusion criteria were included in the study as eligible subjects. Semen tests were conducted on 30 subjects in each group. Subject background is shown in the table below. No significant differences were observed between the groups.

2. Data analysis for primary and secondary endpoints The primary endpoint was the semen test items, and the secondary endpoints were the SF-36, POMS2-AS, and post-intake questionnaire. Changes after the intervention were examined in both groups.
The semen test results are shown in the table below. No significant differences were observed between groups in the amount of change in any of the items.

The table below shows the trends in SF-36, which was set as a secondary endpoint. No significant differences were observed between groups in the amount of change in any of the parameters.

The changes in POMS2-AS scores over time are shown in the table below. No significant differences were observed between groups in the amount of change for any item. No significant differences were observed between groups in the amount of change for any item in the post-ingestion questionnaire.

No significant differences were observed between groups in the amount of change in luteinizing hormone (LH), follicle-stimulating hormone (FSH), or testosterone. In a comparison within the groups, testosterone (at week 12) in the test food group and placebo group increased significantly compared to pre-treatment levels.

3. Subgroup analysis In order to examine male vitality according to age, efficacy analysis subjects were divided into prime-age (35-44 years) and middle-aged (45-65 years) based on their age at the time of the pre-test, and subgroup analysis of semen tests was conducted (Tables 9 and 10). In the prime-age group, the test food group showed a significantly higher change in sperm concentration (sperm count) at 12 weeks compared to the placebo group (Figure 2). In the middle-aged group, no significant differences were observed between groups in the change in any item in the semen test.

Next, an analysis was conducted on subjects aged 40 and over, based on the fact that many people in their 40s responded to the National Health and Nutrition Survey that they were "not getting much" or "not getting any" rest through sleep. As a result, as shown in the table below, the test food group showed a significant increase (improvement) compared to the placebo group in the sleep and eye-related items in the post-ingestion questionnaire, such as "I'm sleeping soundly now" and "I wake up less often to urinate (less frequent urination at night)."

Furthermore, to examine the effect of this study food on preventing lifestyle-related diseases, an analysis was conducted on triglycerides in a group with high triglycerides (TG 150mg/dL or higher) at the pre-test, and on LDL-cholesterol in a group with high LDL-cholesterol (LDL-C 155mg/dL or higher). As a result, no significant differences were observed between groups in the amount of change in any of the items, but it was observed that the test food group had a greater reduction in triglycerides and LDL-cholesterol at 12 weeks than the placebo group.

4. Safety Evaluation A safety evaluation was conducted on 55 safety analysis subjects in each group (Figure 1) for both adverse events and measured and laboratory values. Regarding adverse events, some subjective symptoms were observed during the study period, but all symptoms were mild, and no adverse events were judged to be serious. In addition, the responsible physician determined that there was no relationship to the study food in any of the following cases. In the measured and laboratory values, significant changes were observed in some items when comparing before and after intake in each group, but the responsible physician determined that none of these would pose a clinical problem.

<Safety assessment through overdose testing>
1. Subjects The purpose of this study was to confirm the safety of excessive intake (5x dose) of salmon roe oil and astaxanthin-containing food "ONO-SR/AST." When investigating food safety, it is desirable to conduct and study across a wide range of age groups and genders, so the 5x dose group consisted of 20 subjects to ensure that men and women of each age group, from their 20s to 60s, were included as evenly as possible. For reference, to confirm the safety of 1x and 2x doses, the number of subjects was reduced to 5 subjects each for the 1x and 2x dose groups.

Prior to conducting the study, a consent form was provided to the subjects, the purpose and contents of the study were fully explained, and the subjects' voluntary consent was obtained in writing. Subjects who gave consent were then subjected to pre-examinations (lifestyle questionnaire, medical interview, physical measurements, physical examination, and fasting clinical test). Based on the results of the pre-examinations, subjects who did not meet the following exclusion criteria were selected.

Exclusion criteria: (1) individuals who regularly consume health foods rich in DHA, EPA, or astaxanthin; (2) individuals with or a history of serious illnesses such as diabetes, liver disease, kidney disease, or heart disease; (3) individuals at risk of developing allergies related to the study; (4) individuals currently undergoing treatment for an illness that may affect the study, or individuals with a history of a chronic or serious illness that required medication; (5) individuals with a history or current history of drug or alcohol dependence; (6) individuals who are deemed unsuitable as subjects based on the clinical test values and measurements from the pre-examination; (7) individuals who have participated in other clinical trials within one month of obtaining consent to participate in this study, or individuals who plan to participate in other clinical trials after obtaining consent to participate in this study; (8) individuals who are pregnant, breastfeeding, or planning to become pregnant or breastfeed during the study period; (9) individuals who are deemed unsuitable as subjects based on the results of their responses to the lifestyle questionnaire; (10) individuals who are otherwise deemed unsuitable as subjects by the responsible physician.

2. Test Food The test food "ONO-SR/AST" (Ono Pharmaceutical Co., Ltd.) was a soft capsule containing 1g of salmon roe oil (150mg DHA, 60mg EPA) and 200μg of astaxanthin per 4 capsules, with gelatin, glycerin, water, and caramel coloring as secondary ingredients. The raw materials and nutritional components are shown in the table below.

3. Test method The test was an open-label study. The 1x dose group was given one bag (4 tablets), the 2x dose group two bags (8 tablets), and the 5x dose group five bags (20 tablets) per day, taken without chewing with water or lukewarm water after a meal. The time of intake was not specified, and the drug could be taken in multiple doses throughout the day. The intake period was four weeks. During the test period, participants were instructed not to change their lifestyle habits such as drinking, eating, and sleeping as much as possible, to limit excessive exercise that deviated greatly from the normal range, and to limit dieting and overeating, not to start new exercise, and not to stop previous exercise habits.

　Subjects were asked to visit the hospital in the second and fourth weeks after taking the test food, where they were interviewed (to check their physical condition), measured physically, and had a fasting clinical test. They were also instructed to keep a study diary every day during the study period, which recorded their intake of the test food, changes in their physical condition, mood in daily life, sleep (how well they fell asleep and slept), and medication status. Similar tests were planned to be conducted two weeks after the end of intake, but due to the declaration of a state of emergency due to COVID-19, subjects were not asked to visit the hospital in the second week after the end of intake to ensure their safety. After the end of the intake period, subjects continued to enter information into the diary for two weeks.

4. Test items and evaluation items The tests included a medical interview to check the patient's physical condition, confirmation of the presence or absence of adverse events, physical measurements including height (pre-test only), weight, and BMI, physical examinations including systolic and diastolic blood pressure, pulse rate, fasting clinical hematological tests including white blood cell count, red blood cell count, hemoglobin, hematocrit, and platelet count, blood biochemistry tests including total protein, albumin, total bilirubin, direct bilirubin, indirect bilirubin, ALP, AST, ALT, LD, γ-GT, total cholesterol, triglycerides, HDL-cholesterol, LDL-cholesterol, urea nitrogen, creatinine, uric acid, sodium, potassium, chloride, blood glucose, and HbA1c (pre-test only), and urine tests including protein qualitative, glucose qualitative, and occult blood reaction. Except for items that were only pre-tested, each test was performed at the pre-test, the second week of intake, and the fourth week of intake. In addition, the clinical testing was outsourced to LSI Medience Corporation (Shinagawa-ku, Tokyo).

　Evaluation items were adverse events, measurement values, and test values. Adverse events were subjective symptoms reported in the interview and diary, as well as abnormal fluctuations in test values.

5. Statistical analysis The measured values before and after intake were compared using a one-sample t-test. The significance level of the test was set at 5% on both sides. Microsoft Excel (Microsoft Corporation) was used as the statistical analysis software.

Result 1. Subject background In this study, pre-testing was conducted on 62 subjects who provided written consent to participate in the study. Based on the results of the pre-testing, 15 men and 15 women, a total of 30 subjects, who met the inclusion criteria but did not meet the exclusion criteria, were selected as eligible subjects for this study. Subject background is shown in Table 2. Note that some subjects had clinical test values that exceeded the reference range (test reference values of the clinical testing institution), but in all cases the responsible physician examined each subject individually and, after determining that there were no problems with their participation in the study, they were included in the study.

All 30 subjects began taking the test food, but one man dropped out on the 15th day of intake, and one woman on the first day after the end of intake, due to the spread of COVID-19 infection and the fact that they refrained from going out, and one man dropped out on the first day after the end of intake due to being too busy to continue the study. With these three subjects dropping out, 27 subjects (1x intake group: 5 subjects, 2x intake group: 5 subjects, 5x intake group: 17 subjects) completed the intake of the test food. Although there was a plan to test the subjects two weeks after the end of intake, due to the declaration of a state of emergency due to COVID-19, subjects were not allowed to come to the hospital to ensure their safety. Therefore, in this study, safety was evaluated using measurement and test values from data up to the fourth week. Therefore, the measurement and test values were evaluated for the 27 subjects who completed intake, and all 30 subjects were evaluated for adverse events as analysis subjects.

2. Adverse events There were 13 cases of fluctuations in the measured values and test values for each subject in 9 subjects. None of the fluctuations in the test values were considered to be abnormal (not adverse events).
The incidence and total number of adverse events was 0 out of 5 subjects (0%) at 1x dose, 5 out of 5 subjects (40%) at 2x dose, and 8 out of 20 subjects (25%) at 5x dose. All adverse events were "mild," and no serious adverse events occurred in this study. In addition, the association with the test food was deemed "not present."

3. Measurements and Test Values There were some physical measurements, physical test values, and clinical test values (hematology tests, blood biochemistry tests) that showed significant changes after intake compared to before, but the responsible physician determined that all of these changes were at a level that would not pose clinical problems. Several subjects showed positive clinical test values (urine tests), but as this was thought to be due to the effects of menstruation or temporary fluctuations, the responsible physician determined that there would be no clinical problems.

Conclusion These results show that there are no safety issues when an overdose test is conducted in which subjects take the salmon roe oil and astaxanthin-containing food "ONO-SR/AST" in amounts 1, 2, or 5 times the recommended daily intake amount for 4 consecutive weeks.

Discussion: A study was conducted on men aged 35 to 65 years old to examine the effect of food containing salmon roe oil (DHA 150 mg, EPA 60 mg) on increasing male vitality and the effect on subjective assessment of physical and mental health status when taken continuously for 12 weeks, compared with a placebo. As a result, no significant differences were observed in the primary outcome of semen analysis, or the secondary outcomes of SF-36, POMS2-AS, post-ingestion questionnaire, LH, FSH, and testosterone.

In order to examine the effect on sperm depending on age, a subgroup analysis was conducted based on the prior average age. In the Ministry of Health, Labor and Welfare's health policy "Health Japan 21," issues are extracted and goals are set for 25-44 year olds as the prime age, 45-65 year olds as the middle age, and 65 years and older as the elderly age, and the results of this subgroup analysis showed that sperm concentration (sperm count) was significantly higher in the test food group compared to the placebo group in the prime age group of 35 to 44 years old. These results suggest that the test food increases sperm concentration (sperm count) in prime age subjects.

In the semen tests conducted in this study, the test food group showed a significant increase in sperm concentration compared to the placebo group in the middle-aged subjects. According to the WHO standard values for semen tests, the lower limit for total sperm count is 3900 x ¹⁰⁴ and sperm concentration is 1600 x ¹⁰⁴ /mL, but in this study, the sperm concentration changed from 6373 x ¹⁰⁴ /mL to 6813 x ¹⁰⁴ /mL in the placebo group and from 5605 x ¹⁰⁴ /mL to 9200 x ¹⁰⁴ /mL in the test food group, which is within the normal range.

　Supplementing omega-3 polyunsaturated fatty acids, which cannot be adequately obtained through modern diets, with the test foods and supporting the maintenance of normal sperm counts is thought to lead to the maintenance and improvement of men's reproductive health.

In regards to safety, some subjective symptoms were reported during the study period, but all were mild and the lead physician determined that there was no relationship to the study food. Additionally, no adverse effects were observed in an overdose test of the test food (5 times the amount), demonstrating the safety of the test food.

The present disclosure relates to increasing sperm count and/or sperm concentration and may be used in the medical or food fields.

Claims (8)

1. A composition for increasing sperm count and/or sperm concentration comprising a fish egg lipid preparation.
2. The composition according to claim 1, which is formulated to provide a daily intake of about 100 mg to 750 mg of docosahexaenoic acid (DHA).
3. The composition according to claim 1, which is formulated to provide a daily intake of about 30 mg to 300 mg of eicosapentaenoic acid (EPA).
4. The composition according to claim 1, which is formulated to provide a daily intake of about 100 mg to 750 mg of DHA and about 30 mg to 300 mg of EPA.
5. The composition of claim 1, which is formulated to provide a daily intake of about 160 mg of DHA and about 100 mg of EPA.
6. The composition according to any one of claims 1 to 5, which is taken daily for at least 12 weeks.
7. The composition according to any one of claims 1 to 6, which is to be taken by middle-aged men.
8. The composition according to any one of claims 1 to 7, which is a food composition.

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