Feed composition for improving lipid metabolism comprising lactic acid bacteria fermented Hermetia illucens

Abstract

This specification relates to a feed composition comprising a lactic acid fermented product of a black soldier fly solvent extract, wherein the lactic acid fermented product of the black soldier fly solvent extract has the effect of promoting animal growth, increasing the ratio of beneficial bacteria in the intestines, increasing unsaturated fatty acids in the body, and / or improving lipid metabolism.

Description

Technology Field

[0001] This specification relates to a feed composition comprising a lactic acid fermented product of a black soldier fly solvent extract, and specifically, it can be used for promoting animal growth, increasing the ratio of beneficial bacteria in the intestines, increasing unsaturated fatty acids in the body, and / or improving lipid metabolism. Background Technology

[0002] Recently, as the production burden on aquaculture has increased due to high demand for fishmeal and the resulting price hikes, linkages are being established with the development of insect proteins for aquaculture and livestock farming. The search for alternative and sustainable proteins requires edible solutions in the short term, and the benefits of insects as feed options are being highlighted. In particular, insects serve as a natural food source for various fish and poultry; for instance, chickens peck at worms and larvae in the topsoil and straw, while maggots are used as bait in fishing.

[0003] Major insects used as animal feed include common housefly larvae, silkworms, mealworms, grasshoppers, or termites. While common housefly larvae have been used as a substitute for fishmeal in broiler production, concerns have been raised regarding disease transmission and bacterial proliferation in livestock feed. In the case of termites, although they have been used as food for fish or birds, they are difficult to raise and have raised safety issues due to methanol emissions. Silkworms have been reported to have no impact compared to general feed in terms of feed intake, weight gain, feed efficiency, or protein efficiency. Recently, black soldier flies, capable of decomposing organic substances such as spoiled food or feces, are a protein source that can replace fishmeal and soybean meal, containing 40% protein and 30% fat. The fatty acid composition of black soldier fly larvae generally shows a higher content of saturated fatty acids (SFA; 59.1%) than unsaturated fatty acids (UFA; 37.4%), and among saturated fatty acids, lauric acid (C12:0) has the highest content at 35.72%, accounting for approximately 33% of the total fatty acids (Makkar et al. 2014). Lauric acid is a type of medium-chain fatty acid (MCFA) composed of 6 to 12 carbon atoms and possesses antibacterial properties, so it can be used as a natural antibiotic for gut health (Zeitz et al., 2015).

[0004] Therefore, to utilize insects in general animal feed, it is necessary to ensure safety through heavy metal content and toxicity assessments, prepare for contamination and spoilage caused by microorganisms such as bacteria and viruses, and conduct research on safe long-term storage, distribution, and preservation methods, as well as on feed ratios, growth, and immune enhancement. Meanwhile, in the case of fermentation methods using microorganisms, new efficacy distinct from the existing benefits of the raw materials can be imparted, and numerous harmful components can be transformed into safe ones. Furthermore, by breaking down nutritional components into low-molecular-weight compounds through the action of decomposing enzymes, absorption can be increased, irritation can be reduced, and allergenic components can be decreased.

[0005] There is a high need for research regarding fermented edible insect extracts, which are produced by generating natural antimicrobial substances using bioconversion technology with lactic acid bacteria and applying them to edible insects to enhance antimicrobial, antioxidant, and preservative functions for storage, distribution, and preservation; and regarding fermented edible insect extracts that improve bioavailability and functionality by breaking down amino acids, peptides, fats, and carbohydrates using the enzymes of the lactic acid bacteria themselves. Furthermore, fermentation by beneficial intestinal microorganisms generates short-chain fatty acids such as acetic acid, propionic acid, and butyric acid, which serve as major energy sources for the intestinal microbiome and the human body. Short-chain fatty acids (SCFAs) in the intestines act as nutrients for intestinal epithelial cells, affecting the viable cell count and ratio of the intestinal microbial community, pH, and gene expression for cell proliferation (Cook 1998). Short-chain fatty acids have functions to improve irritable bowel syndrome and inflammatory bowel disease, as well as to prevent cardiovascular disease and obesity (Kumar 2012).

[0006] The masu salmon is a fish belonging to the family Salmonidae, along with trout, coho salmon, and humpback salmon. The scientific name of the masu salmon is Oncorhynchus masou, and currently, the only species of the genus Salmon that inhabit or return to rivers in Korea are the common salmon, masu salmon, chum salmon, and rainbow trout. The chum salmon is the anadromous form (O. masou var masou) of the masu salmon, while the term "masu salmon" commonly refers to the landlocked form (O. masou var ishikawa). The cold-water fish species, the cherry salmon, is an economically viable fish species that is rich in essential amino acids (threonine, valine, methionine, isoleucine, leucine, phenylalanine, lysine, arginine, histidine), vitamin C (600 ug / 100g), and vitamin E (200 ug / 100g), and is also rich in unsaturated fatty acids such as oleic acid (C18:1).

[0007] There has been no research to date on the effects of fermenting black soldier fly, which is rich in long-chain fatty acids, with lactic acid bacteria capable of producing short-chain fatty acids, not only as a protein substitute but also on the improvement of lipid metabolism through changes in animal gut microbiota, growth, and fatty acids, and thus research on this is necessary. The problem to be solved

[0009] Under the background described above, the inventors confirmed the effects of a feed composition containing a fermented black soldier fly lactic acid bacteria on promoting animal growth, increasing the ratio of beneficial bacteria in the intestines, and increasing the content of unsaturated fatty acids in the body, thereby completing the present invention.

[0010] The present specification provides a feed composition comprising a lactic acid fermented product of a black soldier fly solvent extract, and

[0011] The above lactic acid bacteria may be one or more selected from the group consisting of Lactobacillus plantarum, Lactococcus lactis, Leuconostoc mesenteroides, and Lactobacillus brevis, but are not limited thereto.

[0012] The above feed composition may be a feed composition for promoting animal growth, a feed composition for increasing the ratio of beneficial intestinal bacteria, a feed composition for improving lipid metabolism in the animal body, and / or a feed composition for increasing the content of unsaturated fatty acids in the animal body, but is not limited thereto.

[0013] This specification

[0014] (1) A step of preparing black soldier fly extract; and

[0015] (2) A method for preparing a feed composition is provided, comprising the step of inoculating the black soldier fly extract prepared above with lactic acid bacteria and culturing it to produce a fermented product.

[0016] The present specification provides a method for promoting growth in animals, a method for increasing the ratio of beneficial bacteria in the animal's intestines, a method for improving lipid metabolism in the animal's body, and / or a method for increasing the content of unsaturated fatty acids in the animal's body, comprising the step of feeding a feed composition containing a lactic acid fermented product of a black soldier fly solvent extract.

[0017] The above lactic acid bacteria may be one or more selected from the group consisting of Lactobacillus plantarum, Lactococcus lactis, Leuconostoc mesenteroides, and Lactobacillus brevis, but are not limited thereto.

[0018] This specification

[0019] Lactobacillus plantarum strain having accession number KCCM13068P.

[0020] Lactococcus lactis strain having accession number KCCM13066P.

[0021] A strain of Leuconostoc mesenteroides having accession number KCCM13067P; and / or

[0022] Provides a Lactobacillus brevis strain having accession number KCCM13069P. means of solving the problem

[0023] The present specification provides a feed composition comprising a lactic acid fermented product of a black soldier fly solvent extract, and

[0024] The above lactic acid bacteria may be one or more selected from the group consisting of Lactobacillus plantarum, Lactococcus lactis, Leuconostoc mesenteroides, and Lactobacillus brevis, but are not limited thereto.

[0025] The above lactic acid bacteria is

[0026] Lactobacillus plantarum having accession number KCCM13068P;

[0027] Lactococcus lactis having accession number KCCM13066P;

[0028] Leuconostoc mesenteroides having accession number KCCM13067P; and

[0029] It may be one or more lactic acid bacteria selected from the group consisting of Lactobacillus brevis having accession number KCCM13069P, but is not limited thereto.

[0030] The above feed may be intended for use in fish, shrimp, and / or pets, but is not limited thereto.

[0031] The lactic acid fermented product of the above black soldier fly solvent extract may have one or more characteristics selected from the group consisting of the characteristics of (1) to (3) below, but is not limited thereto:

[0032] (1) Growth-promoting ability of animals;

[0033] (2) Increase in the ratio of beneficial gut bacteria; and

[0034] (3) Improvement of lipid metabolism in animals.

[0035] The above feed composition may be a feed composition for promoting animal growth, a feed composition for increasing the ratio of beneficial intestinal bacteria, a composition for improving lipid metabolism in the animal body, and / or a feed composition for increasing the content of unsaturated fatty acids, but is not limited thereto.

[0036] This specification

[0037] (1) A step of preparing black soldier fly extract; and

[0038] (2) A method for preparing a feed composition is provided, comprising the step of inoculating the black soldier fly extract prepared above with lactic acid bacteria and culturing it to produce a fermented product.

[0039] The step of preparing the black soldier fly extract in the method for preparing the above feed composition

[0040] (1) Step of drying black soldier flies;

[0041] (2) A step of grinding the dried black soldier fly to produce powder; and

[0042] (3) The method may include, but is not limited to, a step of mixing the above-prepared powder with a solvent to extract.

[0043] The method for preparing the above feed composition may additionally include, but is not limited to, a step of mixing the prepared fermented product with one or more selected from the group consisting of LT fishmeal (Blumar, Denmark), soybean meal, wheat gluten, tankage meal (Puyer biopharma, China), starch, wheat flour, mineral mix (Puyer biopharma, China), vitamin mix (Puyer biopharma, China), vitamin C, vitamin E, MCP (Monocalcium Phosphate; AAKO, Japan), choline, taurine, and fish oil (Shangdong pharmaceutical, China).

[0045] The present invention will be explained in more detail below.

[0047] In this specification, “animal” refers to a group of organisms corresponding to plants among the two branches of the biological kingdom that consume organic matter as nutrients and have differentiated digestive, excretory, and respiratory organs, and may include, but is not limited to, fish, mammals, reptiles, amphibians, birds, etc.

[0048] In this specification, "black soldier fly" refers to an insect of the family Hermetidae, order Diptera, class Insecta, class Arthropoda. It is characterized by living in dirty water, manure piles, etc., and being distributed in Korea, Japan, and other regions. Its larvae possess excellent organic matter decomposition capabilities and have been industrialized for use in the treatment of food waste. The above "black soldier fly" may refer to one or more selected from the group consisting of eggs, larvae, pupae, and adults of the black soldier fly (Hermetia illucens). In one example, the black soldier fly may be the larva and / or pupa of the black soldier fly, and in another example, it may be the larva of the black soldier fly.

[0049] In this specification, the term “feed” may refer to food given to livestock (animals, etc.), but is not limited thereto.

[0050] Provided in this specification

[0051] Lactobacillus plantarum strain having accession number KCCM13068P.

[0052] Lactococcus lactis strain having accession number KCCM13066P.

[0053] A strain of Leuconostoc mesenteroides having accession number KCCM13067P; and / or

[0054] A strain of Lactobacillus brevis having accession number KCCM13069P is used in the fermentation of black soldier fly extract to produce a fermented product, and said fermented product may be used to promote growth in animals, increase the proportion of beneficial bacteria in the intestines and / or improve lipid metabolism in animals, but is not limited thereto.

[0055] The black soldier fly, which serves as the raw material for the feed additive provided by the present invention, can be freeze-dried and used in powder form, and, if necessary, may undergo additional washing and fasting steps prior to the freeze-drying step. The fasting step is a step for emptying the internal organs of the black soldier fly and may be carried out by stopping the feed supply to the black soldier fly or by feeding glutinous rice flour within the scope of the purpose, but is not limited thereto.

[0056] The above pulverization process can be performed by appropriately utilizing grinding and resting times as needed, and by having a resting time, it has the advantage of minimizing damage to the black soldier fly larva sample caused by heat generated in the grinder. In one embodiment, black soldier fly larva powder (Hermetia illucens; Hi) was prepared by repeating a grinding cycle of grinding for 1 minute followed by resting for 3 minutes five times.

[0057] The above black soldier fly extract can be prepared using black soldier flies, dried black soldier flies, and / or freeze-dried black soldier flies, and if necessary, black soldier flies, dried black soldier flies, and / or freeze-dried black soldier flies can be powdered and used.

[0058] In one embodiment, the black soldier fly extract may be used with an appropriate extraction solvent selected as needed, and the extraction solvent may be one or more selected from the group consisting of alcohols having 1 (C1) to 4 (C4) carbon atoms and water. For example, it may be water, straight-chain or branched-chain alcohols having 1 to 4 carbon atoms, and mixtures thereof; more specifically, it may be ethanol (EtOH) or an aqueous solution of ethanol with an ethanol concentration of 10 to 99 v / v%, 20 to 99 v / v%, 25 to 99 v / v%, 30 to 99 v / v%, 10 to 90 v / v%, 20 to 90 v / v%, 25 to 90 v / v%, 30 to 90 v / v%, 10 to 70 v / v%, 20 to 70 v / v%, 25 to 70 v / v%, or 30 to 70 v / v%. The extraction method for preparing the extract may use a conventional extraction method, for example, extraction with an extraction solvent, supercritical fluid extraction, ultrasonic extraction, hot water extraction, etc., and preferably, a hot water extraction method may be used. The solvent extraction time may be 1 minute to 1 hour, 1 minute to 40 minutes, 1 minute to 30 minutes, 1 minute to 20 minutes, 1 minute to 15 minutes, 5 minutes to 1 hour, 5 minutes to 40 minutes, 5 minutes to 30 minutes, 5 minutes to 20 minutes, 5 minutes to 15 minutes, 10 minutes to 1 hour, 10 minutes to 40 minutes, 10 minutes to 30 minutes, 10 minutes to 20 minutes, 10 minutes to 15 minutes, 14 minutes to 1 hour, 14 minutes to 40 minutes, 14 minutes to 30 minutes, 14 minutes to 20 minutes, or 14 minutes to 15 minutes, for example, 15 minutes, but is not limited thereto.

[0059] In one embodiment, the extraction temperature during solvent extraction is 50 to 200°C, 50 to 160°C, 50 to 150°C, 50 to 140°C, 50 to 130°C, 50 to 125°C, 50 to 121°C, 80 to 200°C, 80 to 160°C, 80 to 150°C, 80 to 140°C, 80 to 130°C, 80 to 125°C, 80 to 121°C, 100 to 200°C, 100 to 160°C, 100 to 150°C, 100 to 140°C, 100 to 130°C, 100 to 125°C, 100 to 121°C, 110 to 200°C, 110 to The temperature may be 160°C, 110 to 150°C, 110 to 140°C, 110 to 130°C, 110 to 125°C, 110 to 121°C, 120 to 200°C, 120 to 160°C, 120 to 150°C, 120 to 140°C, 120 to 130°C, 120 to 125°C, or 120 to 121°C, for example, 121°C, and the solvent may be water, but is not limited thereto.

[0060] In this specification, “extract” may be a hot water extract, solvent extract, distillation extract, ultrasonic extract, press extract, supercritical extract, and / or ultra-high pressure extract, and may be extracted by methods of hot water extraction, solvent extraction, distillation extraction, ultrasonic extraction, press extraction, supercritical extraction, and / or ultra-high pressure extraction, but is not limited thereto.

[0061] The lactic acid fermented product of the black soldier fly extract mentioned above can be used as a raw material for fermentation by using the black soldier fly extract as is, or as a raw material for fermentation after undergoing post-processing steps such as centrifugation or filtration. In order to minimize quantitative loss in order to utilize the active substances of the black soldier fly, the entire black soldier fly extract may be used, or only the supernatant of the black soldier fly extract may be used.

[0062] The lactic acid bacteria fermented product included in the feed composition provided in this specification may be obtained by adding one or more carbon sources selected from the group consisting of dextrose, galactose, mannose, sucrose (sugar, sucrose), glucose (glucose), starch, fructose, lactose (milk sugar), maltose (malt sugar), cellulose, lactulose, trehalose, cellobiose, chitobiose, kozibiose, nigerose, isomaltose, sophorose, laminaribiose, gentiobiose, turanos, maltulose, isomaltulose, gentiobiulose, mannobiose, melivios, beribiulose, rutinose, rutinulose, xylobiose, and rhamnose to the black soldier fly extract and culturing lactic acid bacteria, but is not limited thereto.

[0063] In one embodiment, the carbon source is 0.1 to 10 (w / v)%, 0.1 to 7 (w / v)%, 0.1 to 4 (w / v)%, 0.1 to 3 (w / v)%, 0.1 to 2.3 (w / v)%, 0.1 to 2 (w / v)%, 0.5 to 10 (w / v)%, 0.5 to 7 (w / v)%, 0.5 to 4 (w / v)%, 0.5 to 3 (w / v)%, 0.5 to 2.3 (w / v)%, 0.5 to 2 (w / v)%, 1 to 10 (w / v)%, 1 to 7 (w / v)%, 1 to 4 (w / v)%, 1 to 3 (w / v)%, and 1 to 2.3 based on the volume of the extract. (w / v)%, 1 to 2 (w / v)%, 1.5 to 10 (w / v)%, 1.5 to 7 (w / v)%, 1.5 to 4 (w / v)%, 1.5 to 3 (w / v)%, 1.5 to 2.3 (w / v)%, 1.5 to 2 (w / v)%, 1.7 to 10 (w / v)%, 1.7 to 7 (w / v)%, 1.7 to 4 (w / v)%, 1.7 to 3 (w / v)%, 1.7 to 2.3 (w / v)%, 1.7 to 2 (w / v)%, 2 to 10 (w / v)%, 2 to 7 (w / v)%, 2 to 4 (w / v)%, 2 to 3 (w / v)%, or 2 to It may be added at 2.3 (w / v)%, for example, at 2 (w / v)%, but is not limited thereto.

[0064] The lactic acid bacteria fermentation product included in the feed composition provided in this specification may be obtained by adding one or more nitrogen sources selected from the group consisting of yeast, milk casein, peptone, soy peptone, casein peptone, meat peptone, polypeptone, tryptone, soytone, and beef extract to the black soldier fly extract and culturing lactic acid bacteria, but is not limited thereto, and the yeast may be a yeast extract, specifically, may be in powder form, but is not limited thereto.

[0065] In one embodiment, the nitrogen source is 0.1 to 10 (w / v)%, 0.1 to 5 (w / v)%, 0.1 to 2 (w / v)%, 0.1 to 1 (w / v)%, 0.1 to 0.7 (w / v)%, 0.1 to 0.5 (w / v)%, 0.3 to 10 (w / v)%, 0.3 to 5 (w / v)%, 0.3 to 2 (w / v)%, 0.3 to 1 (w / v)%, 0.3 to 0.7 (w / v)%, 0.3 to 0.5 (w / v)%, 0.4 to 10 (w / v)%, 0.4 to 5 (w / v)%, 0.4 to 2 (w / v)%, and 0.4 to 1 based on the volume of the extract. It may be added in an amount of (w / v)%, 0.4 to 0.7 (w / v)%, 0.4 to 0.5 (w / v)%, 0.5 to 10 (w / v)%, 0.5 to 5 (w / v)%, 0.5 to 2 (w / v)%, 0.5 to 1 (w / v)%, or 0.5 to 0.7 (w / v)%, for example, 0.5 (w / v)%, but is not limited thereto.

[0066] In the feed additive and / or feed composition comprising a lactic acid fermented product of black soldier fly extract provided in this specification, the lactic acid fermented product of black soldier fly extract may be prepared by undergoing the step of adding (inoculating) one or more types of lactic acid bacteria to the lactic acid fermented product of black soldier fly extract and fermenting it.

[0067] In one embodiment, the lactic acid bacteria may be one or more types of lactic acid bacteria selected from the group consisting of Lactobacillus plantarum, Lactococcus lactis, Leuconostoc mesenteroides, and Lactobacillus brevis, but is not limited thereto.

[0068] In one example, the above lactic acid bacteria is

[0069] Lactobacillus plantarum having accession number KCCM13068P;

[0070] Lactococcus lactis having accession number KCCM13066P;

[0071] Leuconostoc mesenteroides having accession number KCCM13067P; and

[0072] It may be one or more lactic acid bacteria selected from the group consisting of Lactobacillus brevis having accession number KCCM13069P, but is not limited thereto. The above bacteria may be obtained and used from the Korean Culture Center of Microorganisms (KCCM).

[0073] The above Lactobacillus plantarum strain may be a strain having a 16S rRNA gene having sequence identity of 99% or more, 99.5% or more, or 99.9% or more with the nucleotide sequence of SEQ ID NO. 3, for example, a strain having a 16S rRNA sequence consisting of the nucleotide sequence of SEQ ID NO. 3, and more preferably, a strain of accession number KCCM13068P.

[0074] The above Lactococcus lactis strain may be a strain having a 16S rRNA gene having sequence identity of 99% or more, 99.5% or more, or 99.9% or more with the nucleotide sequence of SEQ ID NO. 4, for example, a strain having a 16S rRNA sequence consisting of the nucleotide sequence of SEQ ID NO. 4, and more preferably, a strain with accession number KCCM13066P.

[0075] The above-mentioned Leuconostoc mesenteroides strain may be a strain comprising a 16S rRNA gene having sequence identity of 99% or more, 99.5% or more, or 99.9% or more with the nucleotide sequence of SEQ ID NO. 5, for example, a strain comprising a 16S rRNA sequence consisting of the nucleotide sequence of SEQ ID NO. 5, and more preferably, a strain of accession number KCCM13067P.

[0076] The above Lactobacillus brevis strain may be a strain having a 16S rRNA gene having sequence identity of 99% or more, 99.5% or more, or 99.9% or more with the nucleotide sequence of SEQ ID NO. 6, for example, a strain having a 16S rRNA sequence consisting of the nucleotide sequence of SEQ ID NO. 6, and more preferably, a strain of accession number KCCM13069P.

[0077] In the feed additive and / or feed composition comprising a lactic acid fermented product of black soldier fly extract provided by the present invention, the feed may be a feed for use in fish, crustaceans, and / or animals, and the fish may be one or more selected from the group consisting of trout, spotted flounder, stone flounder, halibut, rockfish, rainbow trout, yellowtail, eel, carp, ornamental fish, catfish, herring, sea bass, cod, anglerfish, pufferfish, scorpionfish, horse mackerel, skate, smelt, etc., the crustaceans may be shrimp, etc., and the animals may be one or more selected from the group consisting of reptiles, amphibians, chickens (e.g., broiler chickens, laying hens, etc.), pigs, cattle, goats, sheep, horses, dogs, cats, ducks, geese, etc. In one example, the above feed may be applied to fish and / or shrimp, and / or to pets such as dogs or cats, but is not limited thereto.

[0078] In a feed additive comprising a lactic acid fermented product of black soldier fly extract provided by the present invention, the content of the lactic acid fermented product of the black soldier fly extract is 0.1 to 50 wt%, 0.1 to 40 wt%, 0.1 to 30 wt%, 0.1 to 20 wt%, 0.1 to 10 wt%, 0.1 to 5 wt%, 1 to 50 wt%, 1 to 40 wt%, 1 to 30 wt%, 1 to 20 wt%, 1 to 10 wt%, 1 to 5 wt%, 1 to 4.5 wt%, 2 to 50 wt%, 2 to 40 wt%, 2 to 30 wt%, 2 to 20 wt%, 2 to 10 wt%, 2 to 5 wt%, 2 to 4.5 wt%, 4 to 50 wt%, 4 to 40 wt%, 4 to It may be 30 wt%, 4 to 20 wt%, 4 to 10 wt%, 4 to 5 wt%, 4 to 4.5 wt%, 4.5 to 50 wt%, 4.5 to 40 wt%, 4.5 to 30 wt%, 4.5 to 20 wt%, 4.5 to 10 wt%, or 4.5 to 5 wt%, for example, 4.5 wt%, but is not limited thereto.

[0079] The lactic acid fermented product of the black soldier fly solvent extract of the feed composition provided in this specification may have one or more characteristics selected from the group consisting of the characteristics of (1) to (3) below, but is not limited thereto:

[0080] (1) Growth-promoting ability of animals;

[0081] (2) Increase in the ratio of beneficial gut bacteria; and

[0082] (3) Improvement of lipid metabolism in animals.

[0083] The improvement of lipid metabolism in the animal body described above refers to the synthesis and breakdown (oxidation) of triglycerides, phospholipids, cholesterol, fatty acids, etc., and / or the prevention of obesity, pancreatitis (pancreatic cancer), and dyslipidemia through the management of triglycerides in the body. Specifically, it may refer to the enhancement or synthesis of unsaturated fatty acid content, particularly the enhancement of the content of essential fatty acids such as omega-3 (EPA, DHA, α-linoleic acid), omega-6 (ETA, linolenic acid, arachidonic acid), and / or omega-9, but is not limited thereto.

[0084] In one embodiment, when the above feed composition is consumed, compared to when a feed not containing the black soldier fly solvent extract (hereinafter referred to as "general feed") is consumed, the daily growth rate calculated by the method of [Mathematical Formula 2] measured at the 4th week after consumption of the above feed composition is 1.01 to 3 times, 1.01 to 2 times, 1.01 to 1.5 times, 1.01 to 1.2 times, 1.01 to 1.1 times, 1.01 to 1.08 times, 1.04 to 3 times, 1.04 to 2 times, 1.04 to 1.5 times, 1.04 to 1.2 times, 1.04 to 1.1 times, 1.04 to 1.08 times, 1.06 to 3 times, 1.06 to 2 times, 1.06 to 1.5 times, 1.06 to It may increase by 1.2 times, 1.06 to 1.1 times, 1.06 to 1.08 times, 1.07 to 3 times, 1.07 to 2 times, 1.07 to 1.5 times, 1.07 to 1.2 times, 1.07 to 1.1 times, or 1.07 to 1.08 times, for example, it may increase by 1.076 times, but is not limited thereto.

[0085] [Mathematical Formula 2]

[0086]

[0087] The lactic acid fermented product of black soldier fly extract and the feed additive and / or feed composition comprising the lactic acid fermented product of black soldier fly extract of the present specification may have the effect of increasing the ratio of beneficial bacteria in the intestines.

[0088] In one embodiment, the beneficial intestinal bacteria may be one or more selected from the group consisting of Lactococcus lactis, Enterococcus faecium, Lactobacillus plantarum, Leuconostoc mesentroides, and Lactobacillus brevis, but are not limited thereto.

[0089] In one embodiment, the proportion of Lactococcus lactis in the gut can be increased to 50% or more, 55% or more, 60% or more, 61% or more, e.g., 61.8%, and can be increased to 99.9% or less, 99% or less, but is not limited thereto.

[0090] In one embodiment, a group of animals fed a feed additive containing a lactic acid fermented product of black soldier fly extract and / or a feed composition of the present invention, and a group of animals fed a feed not containing the lactic acid fermented product of black soldier fly extract (hereinafter referred to as "general feed"), at the 4th week after the start of feeding, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus fermentum, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactococcus lactis, Enterococcus faecium, Enterococcus faecalis, Streptococcus thermophilus, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animalis, The distribution of 21 species of intestinal microorganisms, including Lactobacillus brevis and Leuconostoc mesentroides, was confirmed using Illumina MiSeq.

[0091] As a result, it was confirmed that 5 out of 21 types of intestinal microorganisms were distributed at the 4th week after the start of feeding of animals fed a feed additive containing a lactic acid fermented product of black soldier fly extract and / or the feed composition of the present specification. Specifically, Lactococcus lactis, a lactic acid bacterium administered to HiLAB, accounted for the majority at 61.8%, Enterococcus faecium accounted for 32.1%, and the remaining 3 types were Lactobacillus plantarum (0.6%), Leuconostoc mesentroides (4.6%), and Lactobacillus brevis (0.9%), which were administered as notified bacteria.

[0092] The feed composition comprising a lactic acid fermented product of black soldier fly extract provided in this specification may have the effect of increasing the unsaturated fatty acid content in the animal body, but is not limited thereto.

[0093] In one embodiment, the unsaturated fatty acid may be one or more selected from the group consisting of myristoleic acid, palmitoleic acid, oleic acid, linoleic acid, α-linolenic acid, gadoleic acid, eicosatrienoic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, and nervonic acid, but is not limited thereto.

[0094] In one embodiment, the unsaturated fatty acid may be one or more selected from the group consisting of omega-3 unsaturated fatty acids, omega-6 unsaturated fatty acids and omega-9 unsaturated fatty acids, but is not limited thereto.

[0095] In the feed composition comprising a lactic acid fermented product of black soldier fly extract provided in this specification, the feed may be fed alone, or mixed with and / or alternately fed with a general feed that does not contain the lactic acid fermented product of the black soldier fly solvent extract. The feeding amount and frequency of the feed may be appropriately adjusted as needed by a person skilled in the art, depending on the type of animal, the number of individuals, and the age and / or size of the animal.

[0096] In one embodiment, the characteristics of the feed composition containing the lactic acid bacteria fermented product of the black soldier fly extract—(1) growth-promoting ability of animals, (2) increase in the ratio of beneficial bacteria in the intestines, and / or (3) increase in the content of unsaturated fatty acids in the animal body—can be tested by feeding the feed to fish and raising them, but are not limited thereto.

[0097] Feeding the fish can be done by preparing two groups of tanks and placing the same number of fish (e.g., 50 fish) in each group of tanks, and feeding each tank with a feed composition containing a lactic acid fermented product of black soldier fly extract (experimental group) and a feed composition containing soybean meal instead of the fermented product (control group) for 4 weeks, and the rearing conditions of the fish can be created by supplying clean tap water to the rearing tanks at a constant rate and allowing any excess to drain out through the drain, but is not limited thereto. Effects of the invention

[0098] The present invention relates to a feed composition comprising a lactic acid bacteria fermented product of black soldier fly extract, wherein the lactic acid bacteria fermented product has the effect of improving lipid metabolism by promoting animal growth, increasing the ratio of beneficial bacteria in the intestines, and increasing the content of unsaturated fatty acids in the animal body. Brief explanation of the drawing

[0099] Figure 1 is a graph showing LAB (Lactic Acid Bacteria) Counts (log cfu / ml) measurements, which represent the number of colonies formed according to culture conditions and the type of lactic acid bacteria added during culture. Figure 2 is a graph showing the daily growth rate of cherry salmon measured according to the type of feed given. HiLAB is the experimental group fed with feed containing lactic acid fermented black soldier fly larvae extract, and Control is the control group fed with feed not containing lactic acid fermented black soldier fly larvae extract. Figure 3 is a graph showing the distribution of intestinal microbial strains of mountain trout measured according to the type of feed administered. HiLAB is the experimental group fed with feed containing a lactic acid fermented product of black soldier fly larvae extract, and Control is the control group fed with feed not containing a lactic acid fermented product of black soldier fly larvae extract. Figure 4 is a graph showing the content of saturated fatty acids (SFA) and unsaturated fatty acids (UFA) in the body of cherry salmon measured according to the type of feed fed. HiLAB refers to the experimental group fed with feed containing lactic acid fermented black soldier fly larvae extract, and Control refers to the control group fed with feed not containing lactic acid fermented black soldier fly larvae extract. Figure 5 is a graph showing the content of monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFA) in the body of cherry salmon measured according to the type of feed fed. HiLAB refers to the experimental group fed with feed containing lactic acid fermented black soldier fly larvae extract, and Control refers to the control group fed with feed not containing lactic acid fermented black soldier fly larvae extract. Figure 6 is a graph showing the content of omega-3, omega-6, and omega-9 related unsaturated fatty acids in the body of cherry salmon measured according to the type of feed fed. HiLAB refers to the experimental group fed with feed containing lactic acid fermented black soldier fly larvae extract, and Control refers to the control group fed with feed not containing lactic acid fermented black soldier fly larvae extract. Specific details for implementing the invention

[0100] The present invention will be explained in more detail below through examples. However, the following examples are intended to illustrate the present invention and do not limit the scope of the present invention.

[0102] Example 1. Preparation of Black Soldier Fly Extract

[0103] Example 1-1. Preparation of Black Soldier Fly Powder

[0104] Black soldier fly larvae were raised for 15 to 18 hours at a temperature of 27°C and a humidity of 60%, and then fasted for 3 days to clean the intestines of the black soldier fly larvae. Next, the black soldier fly larvae were washed under running water and then treated with insecticide by hot-air drying at 100°C for 24 hours.

[0105] The dried black soldier fly larvae were ground into powder using a grinder. Specifically, to prevent heat generation from the use of the grinder, a grinding cycle of grinding for 1 minute followed by a rest for 3 minutes was repeated 5 times to produce black soldier fly larva powder (Hermetia illucens; Hi).

[0106] Example 1-2. Preparation of Black Soldier Fly Hot Water Extract

[0107] 50g of black soldier fly larva powder (Hi) prepared in Example 1-1 above was mixed with 950ml of water and mixed by irradiating with ultrasound at 750 watts, 10kHz conditions at 10-second intervals for 5 minutes (Sonics Vibra-Cell, USA). Afterwards, the mixture was treated in a 121℃ high-temperature high-pressure autoclave (steam sterilizer) for 5 to 20 minutes to prepare a hot water extract of black soldier fly larva (Hotwater Extract of Hermetia illucens, HiHe). The hot water extract was stored at -70℃ and then freeze-dried, or freeze-dried immediately after preparation to form a powder, or the liquid extract was taken and used in the following experiments.

[0109] Example 2. Isolation and identification of lactic acid bacteria for the preparation of fermented product

[0110] Example 2-1. Isolation of lactic acid bacteria

[0111] To prepare a fermented product using effective strains for fish feed, Lactobacillus plantarum (KCCM13068P), Lactococcus lactis (KCCM13066P), Leuconostoc mesenteroides (KCCM13067P), and Lactobacillus brevis (KCCM13069P), which are lactic acid bacteria strains with functions such as antibacterial and enzymatic activity, were isolated and identified from seawater and used, respectively. The above-mentioned Lactobacillus plantarum (KCCM13068P), Lactococcus lactis (KCCM13066P), Leuconostoc mesenteroides (KCCM13067P), and Lactobacillus brevis (KCCM13069P) were isolated from deep sea water at an aquaculture farm of the Gyeongbuk Fisheries Research Institute in June 2018. The strains isolated from the above-mentioned deep sea water were plated onto DE MAN, ROGOSA and SHARPE (MRS) agar medium supplemented with 0.1% (v / v) bromocresol purple (BCP) and cultured at 37°C for 1 day. Subsequently, single species were isolated by repeatedly selecting yellow colonies formed by reaction with BCP.

[0112] The four isolated strains were cultured in MRS liquid medium, mixed with 50% (v / v) glycerol to achieve a final glycerol concentration of 25%, and then stored at -70℃.

[0113] Example 2-2. Identification of Lactic Acid Bacteria

[0114] The strain isolated in Example 2-1 above was identified using 16S rRNA sequencing analysis. To identify the four strains, the genomic DNA of each strain was first isolated using a mini genomic DNA extraction kit (Promega).

[0115] Using the isolated genomic DNA above as a template, the 16S rRNA gene regions of the four strains above were amplified using a TaKaRa PCR Thermal Cycler (Japan) with the following sets of 27F (Sequence No. 1) and 1492R primers (Sequence No. 2) and the AccPower PCR premix kit (Bioneer, Korea), and the information of the primer sets is shown in Table 1 below.

[0116] primer Nucleotide sequence (5' → 3') Sequence number 27F primer AGAGTTTGATCMTGGCTCAG 1 1492R primer GGTTACCTTGTTACGACTT 2

[0117] The above PCR product was purified by PCR clean-up Gel extraction (Macherey Nagel, USA) and the gene sequence was analyzed using an ABI3730 DNA analyzer (Applied Bioxyxtems, USA). The 16S rDNA sequences of the Lactobacillus plantarum, Lactococcus lactis, Leuconostoc mesenteroides, and Lactobacillus brevis strains isolated in Example 2-1 are shown in SEQ ID NOs 3, 4, 5, and 6, respectively, and the information is shown in Table 2 below.

[0118] strain Nucleotide sequence (5' → 3') Sequence number Lactobacillus plantarum MIRE_TS55 GGGCGAAGTGCCGGGGGTGCCTGATACATGCAAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTAGGAACCAGCCGCTTAAGTGTACAGGATTCC 3 Lactococcus lactis MIRE_P1 TGCCAGGGGGCGGCATGCTATACATGCAAGTTGAGCGCTGAAGGTTGGTACTTGTACCGACTGGATGAGCAGCGAACGGGTGAGTAACGCGTGGGGAATCTGCCTTTGAGCGGGGGACAACATTTGGAAACGAATGCTAATACCGCATAAAAACTTTAAACACAAGTTTTAAGTTTGAAAGATGCAATTGCATCACTCAAAGATGATCCCGCGTTGTATTAGCTAGTTGGTGAGGTAAAGGCTCACCAAGGCGATGATACATAGCCGACCTGAGAGGGTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGACGAAAGTCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAACTCTGTTGGTAGAGAAGAACGTTGGTGAGAGTGGAAAGCTCATCAAGTGACGGTAACTACCCAGAAAGGGACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGTGGTTTATTAAGTCTGGTGTAAAAGGCAGTGGCTCAACCATTGTATGCATTGGAAACTGGTAGACTTGAGTGCAGGAGAGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGCCTGTAACTGACACTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGATGTAGGGAGCTATAAGTTCTCTGTATCGCAGCTAACGCAATAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATACTCGTGCTATTCCTAGAGATAGGAAGTTCCTTCGGGACACGGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCCATCATTAAGTTGGGCACTCTAACGAGACTGCCGGTGATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTCGCGAGACAGTGATGTTTAGCTAATCTCTTAAAACCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGGGAGTTGGGAGTACCCGAAGTAGGTTGCCTAACCGCAAGGAGGGCGCTCCTAAGGTAGACCGCGTGGA 4 Leuconostoc mesenteroides MIRE_ts01 GGGGAAGTGCCGGCGTGCTATACATGCAGTCGAACGCACAGCGAAAGGTGCTTGCACCTTTCAAGTGAGTGGCGAACGGGTGAGTAACACGTGGACAACCTGCCTCAAGGCTGGGGATAACATTTGGAAACAGATGCTAATACCGAATAAAACTTAGTGTCGCATGACAAAAAGTTAAAAGGCGCTTCGGCGTCACCTAGAGATGGATCCGCGGTGCATTAGTTAGTTGGTGGGGTAAAGGCCTACCAAGACAATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCTGCAGTAGGGAATCTTCCACAATGGGCGAAAGCCTGATGGAGCAACGCCGCGTGTGTGATGAAGGCTTTCGGGTCGTAAAGCACTGTTGTATGGGAAGAACAGCTAGAATAGGAAATGATTTTAGTTTGACGGTACCATACCAGAAAGGGACGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGTCCCGAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGACGGTTTATTAAGTCTGATGTGAAAGCCCGGAGCTCAACTCCGGAATGGCATTGGAAACTGGTTAACTTGAGTGCAGTAGAGGTAAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTTACTGGACTGCAACTGACGTTGAGGCTCGAAAGTGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCACACCGTAAACGATGAACACTAGGTGTTAGGAGGTTTCCGCCTCTTAGTGCCGAAGCTAACGCATTAAGTGTTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTTTGAAGCTTTTAGAGATAGAAGTGTTCTCTTCGGAGACAAAGTGACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTGTTAGTTGCCAGCATTCAGATGGGCACTCTAGCGAGACTGCCGGTGACAAACCGGAGGAAGGCGGGGACGACGTCAGATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGCGTATACAACGAGTTGCCAACCCGCGAGGGTGAGCTAATCTCTTAAAGTACGTCTCAGTTCGGATTGTAGTCTGCAACTCGACTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTTTGTAATGCCCAAAGCCGGTGGCCTAACCTTTAGGAAGGAGCCGTCTAAGCAGACAGTCCTCGA 5 Lactobacillus brevis MIRE_TS66 CGGGAATGGGGGGCTGCCGTATACATGCAAGTCGAACGAGCTTCCGTTGAATGACGTGCTTGCACTGATTTCAACAATGAAGCGAGTGGCGAACTGGTGAGTAACACGTGGGGAATCTGCCCAGAAGCAGGGGATAACACTTGGAAACAGGTGCTAATACCGTATAACAACAAAATCCGCATGGATTTTGTTTGAAAGGTGGCTTCGGCTATCACTTCTGGATGATCCCGCGGCGTATTAGTTAGTTGGTGAGGTAAAGGCCCACCAAGACGATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAATGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACACCTTTGAGAGTAACTGTTCAAGGGTTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTTAACCGGAGAAGTGCATCGGAAACTGGGAGACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTAGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATCTTCTGCCAATCTTAGAGATAAGACGTTCCCTTCGGGGACAGAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTCAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTACAACGAGTTGCGAAGTCGTGAGGCTAAGCTAATCTCTTAAAGCCGTTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGCCGTTGAGATAACCTTCGGGAGTCAGCAGTCTAAGTGACAGGGGATCGCCG 6

[0119] 16S rRNA sequence similarity analysis was performed using the 16S rRNA sequence information of each isolated strain above, and the results are shown in Table 3 below.

[0120] Lactic acid bacteria origin isolated strain Proximate 16S rRNA sequence identity Trustee number sea ​​water Lactobacillus plantarum MIRE_TS55(16S rRNA of sequence number 3) Lactobacillus plantarum MT597760.1 (Sequence No. 7) 99% KCCM13068P sea ​​water Lactococcus lactis MIRE_P01(16S rRNA of sequence number 4) Lactococcus lactis MT473540.1 (Sequence No. 8) 99% KCCM13066P sea ​​water Leuconostoc mesenteroides MIRE_TS01 (16S rRNA of sequence number 5) Leuconostoc mesenteroides KC108669.1 (Sequence No. 9) 99% KCCM13067P sea ​​water Lactobacillus brevis MIRE_TS66 (16S rRNA of sequence number 6) Lactobacillus brevis MT597799.1 (Sequence No. 10) 99% KCCM13069P

[0121] As shown in Table 3 above, the lactic acid bacteria Lactobacillus plantarum, Lactococcus lactis, Leuconostoc mesenteroides, and Lactobacillus brevis isolated from seawater in Example 2-1 showed 99% sequence similarity to the Lactobacillus plantarum strain (Accession No. GenBank accession no. MT597760.1) and Lactococcus lactis strain (Accession No. GenBank accession no. GQ337880.1), and the Leuconostoc mesenteroides strain (Accession No. GenBank accession no. MT416441.1) and Lactobacillus brevis strain (Accession No. GenBank accession no. MT597799.1), respectively, based on 16S rRNA analysis results. The strains isolated from the seawater were registered with the Korean Culture Collection Center (KCCM) in 2021 They were deposited on October 27 and received accession number KCCM13068P for Lactobacillus plantarum (MIRE_TS55), accession number KCCM13066P for Lactococcus lactis (MIRE_P1), accession number KCCM13067P for Leuconostoc mesenteroides (MIRE_ts01), and accession number KCCM13069P for Lactobacillus brevis (MIRE_TS66).

[0123] Example 3. Confirmation of optimal conditions for the preparation of black soldier fly extract lactic acid bacteria fermented product

[0124] In preparing a lactic acid fermented product of black soldier fly larvae extract, an experiment was conducted to determine whether the addition of a carbon source is necessary as an optimal condition to effectively induce lactic acid fermentation. To verify this, a medium (Test Medium 1) was prepared by adding 1 L of the hot water extract of black soldier fly larvae (HiHe) prepared in Examples 1-2 above, 2 (w / v)% of galactose (Galactose Sigma, USA) as a carbon source and 0.5 (w / v)% of yeast powder (Kisan Bio, Korea) as a nitrogen source based on the volume of the extract. In addition, as a control, a DE MAN, ROGOSA and SHARPE (MRS) medium, which is a commonly used lactic acid bacteria medium (Control Medium), and a medium containing only 1 L of HiHe (Test Medium 2) were prepared.

[0125] In the three media prepared above (Test Medium 1, Test Medium 2, and Control Medium), 1 x 10⁶ of the four types of lactic acid bacteria—Lactobacillus plantarum (KCCM13068P), Lactococcus lactis (KCCM13066P), Leuconostoc mesenteroides (KCCM13067P), and Lactobacillus brevis (KCCM13069P)—were each added. 8 1 mL aliquots were inoculated at a concentration of cfu / ml, and additionally, 1 x 10 of the above four types of lactic acid bacteria were added. 8 0.25 mL of a mixture was inoculated at a concentration of cfu / ml, and the lactic acid bacteria inoculated into the above medium were cultured at 30°C for 24 hours.

[0126] The optimal culture conditions for the fermented product were determined by measuring the LAB (Lactic Acid Bacteria) Counts (log cfu / ml) after 24 hours of culture. Specifically, the fermented product was diluted to 1 (v / v)% in a solid medium prepared by adding agarose (BD Difco, USA) to each liquid medium, 1 ml was spread onto a LAB plate, and the number of colonies formed was measured after incubation at 30°C for 24 hours. The results are shown in Table 4 and Figure 1 below.

[0127] culture medium Probiotics LAB Counts (log cfu / ml) Control medium Lactobacillus plantarum 6.26E+09 Lactococcus lactis 1.2E+11 Leuconostoc mesenteroides 3.E+11 Lactobacillus brevis 2.E+10 Mixed inoculation of 4 types of probiotics 2.4E+11 Test medium 2 Lactobacillus plantarum 8.E+09 Lactococcus lactis 1.E+11 Leuconostoc mesenteroides 3.E+11 Lactobacillus brevis 4.E+08 Mixed inoculation of 4 types of probiotics 1.2E+10 Test medium 1 Lactobacillus plantarum 2.7E+12 Lactococcus lactis 1.3E+11 Leuconostoc mesenteroides 1.5E+14 Lactobacillus brevis 2.31E+11 Mixed inoculation of 4 types of probiotics 1.4E+13

[0128] As shown in Table 4 above, Test Medium 2 containing a hot water extract of black soldier fly larvae and Test Medium 1, which added 0.5% yeast and 2% galactose to the same, both increased the growth of four types of lactic acid bacteria—Lactobacillus plantarum, Lactococcus lactis, Leuconostoc mesenteroides, and Lactobacillus brevis—compared to the control medium, with Test Medium 1 showing a particularly high degree of increase. In addition, it was confirmed that culturing a mixture of the four types of lactic acid bacteria increased the growth of the bacteria compared to single inoculation.

[0130] Example 4. Confirmation of the effect of black soldier fly fermentation product on fish growth

[0131] Example 4-1. Preparation of fish feed

[0132] An experiment was conducted after manufacturing cherry salmon feed to determine the effect of lactic acid fermentation of black soldier fly extract on fish growth.

[0133] Specifically, a lactic acid bacteria fermentation product (HiLAB) prepared by inoculating a mixture of the four types of lactic acid bacteria into an extract to which 2 (w / v)% of galactose (D-glucose, Galactose Sigma, USA) and 0.5 (w / v)% of yeast powder (Kisan Bio, Korea) were added based on the volume of the extract to the hot water extract of black soldier fly larvae (HiHe) prepared in Examples 1-2 above, and LT fishmeal (Blumar, Denmark), soybean meal, wheat gluten, tankage meal (Puyer biopharma, China), starch, wheat flour, mineral mix (Puyer biopharma, China), vitamin mix (Puyer biopharma, China), vitamin C, vitamin E, MCP (Monocalcium Phosphate; AAKO, Japan), choline, taurine, and fish oil; A feed for mountain trout (experimental feed) was prepared by mixing Shangdong Pharmaceutical (China), and the specific composition information is shown in Table 5 below. As a control group, a feed (control feed) mixed with soybean meal was prepared in place of the lactic acid fermented black soldier fly extract included in the experimental feed, and the specific composition information of the control group is also shown in Table 5 below. The above experimental feed and control feed were prepared by mixing at the Feed Center of the National Institute of Fisheries Science.

[0134] feed ingredients Control feed (weight%) Experimental feed (weight%) LT fishmeal 40.00 40.00 Lactic acid fermented product of black soldier fly extract (HiLAB) 0.00 4.50 Soybean meal 4.50 0 Wheat gluten 12.00 12.00 Tankage Mill 12.00 12.00 starch 6.70 7.20 flour 9.00 9.00 Vitamin Mix 1.00 1.00 Mineral mix 1.00 1.00 Vitamin C 0.20 0.20 Vitamin E 0.10 0.10 MCP 0.50 0.50 Choline 2.50 2.50 Taurine 0.50 0.50 fish oil 10.00 9.50 Total 100.00 100.00

[0135] In addition, component analysis was performed on the above-mentioned feed in a dry state.

[0136] Specifically, 40g of refined sea sand of 20 mesh was mixed with 20g of the above-prepared feed and dried in a dry oven at 105℃ for 5 hours. The moisture content ((weight of feed after drying / weight of feed before drying) x 100) was determined by the atmospheric pressure heating drying method, the ash content was determined by the direct incineration method (drying at 550℃ for 3 hours), the crude protein content was determined by the nitrogen quantification method using an automatic crude protein analyzer (Auto kjeldahl system; Bunchi, Switzerland), and the crude fat content was determined by the ether extraction method. The carbohydrate content was calculated from the analyzed moisture, ash, crude protein, and crude fat content using the following mathematical formula 1, and the analysis results are shown in Table 6 below.

[0137] [Mathematical Formula 1]

[0138] Carbohydrate content (%) = {100% - (Moisture% measured above + Ash% measured above + Crude protein% measured above + Crude fat% measured above}

[0139] control feed Experimental feed Moisture ((w / w)%) 2.86 2.45 Crude protein ((w / w)%) 52.67 52.48 Crude lipid ((w / w)%) 17.21 16.93 Views (crude ash) ((w / w)%) 10.13 10.67 Energy (cal / g) 5,245 5,229 Crude fiber ((w / w)%) 0.13 0.20

[0140] Example 4-2. Conduct of feed feeding and growth experiment on mountain trout

[0141] As the experimental subjects, juvenile individuals of cherry salmon produced by the Gyeongsangbuk-do Freshwater Fish Research Center, acclimated to compound feed (dominant), and raised to a body weight of 14 to 15 g were used. A total of 6 tanks were prepared, and 50 fish were placed in each tank. The feed prepared in Example 4-1 was divided into two groups of three tanks each, and the tanks in each group were fed the control feed (control group) and the experimental feed (experimental group) respectively for 4 weeks. During the experiment, clean tap water was supplied consistently to the rearing tanks, and measures were taken to ensure that any excess water drained out through the drain.

[0142] As described above, while feeding, the total body weight was measured before the start of feeding (Day 0) and at weeks 2 and 4 after the start of feeding. Specifically, for each measurement round, the weight was measured for 50 cherry salmon per tank, and the average value for each group was calculated.

[0143] In addition, to measure the growth rate of the cherry salmon, the daily growth rate (Specific Growth Rate, SGR) was calculated using the method of Equation 2 below.

[0144] [Mathematical Formula 2]

[0145]

[0146] In the above mathematical formula 2, the initial fish body weight refers to the average body weight of the corresponding group of trout immediately before the feed feeding experiment, and the final fish body weight refers to the average body weight of the corresponding group of trout measured at the beginning (week 0 of the experiment start; before feeding the control feed and experimental feed), week 2 (week 2 of the experiment start; week 2 after feeding the control feed and experimental feed), or week 4 (week 4 of the experiment start; week 4 after feeding the control feed and experimental feed), and the results of the daily growth rate measurement at week 4 are shown in Figure 2.

[0147] As shown in Figure 2, the daily growth rate of the experimental group fed with HiLAB was measured to be higher, and it was confirmed that the lactic acid fermented product of black soldier fly larva extract (HiLAB) is effective for fish growth after undergoing the lactic acid fermentation process.

[0149] Example 5. Confirmation of intestinal microorganisms by ingestion of fermented black soldier fly extract

[0150] To confirm the intestinal microbial distribution at week 4 after the start of feeding for the control and experimental groups of Example 4-2 above, the intestines were aseptically excised from 9 trout of each group, washed three times with 0.75 (v / v)% physiological saline, and after removing intestinal contents resembling excrement, only the intestinal mucus was collected in a 1.5 mL tube and stored at -70°C. Each collected sample was incubated at 37°C for 2 days in a test tube containing tryptomen soya broth (TSB; Merck, Germany). 2 mL of the cultured sample was centrifuged at 9,000 rpm for 10 minutes to concentrate the bacteria. The concentrated bacterial genomic DNA was extracted using the QIAamp DNA Mini Kit (Qiagen, Italy). The extracted genomic DNA was diluted to a final concentration of 5 ng / μl, and a 16S rRNA library was constructed using 16S metagenomic Sequencing Library Preparation (Illumina). To amplify the 16S rDNA of bacteria, PCR was performed using Bakt_341F and Bakt_805R primers, which can identify most bacteria, to identify the V1-V2 (340-370 bp) and V3-V4 (440-460 bp) regions, and the primer information is shown in Table 7 below.

[0151] primer Nucleotide sequence (5' → 3') Sequence number Bakt_341F CCTACGGGNGGCWGCAG 11 Bakt_805R GACTACHVGGGTATCTAATCC 12

[0152] (In Table 7 above, N represents an arbitrary sequence as A, G, C, or T / U, W represents A or T / U, H represents A, C, or T / U, and V represents A, G, or C.) PCR was performed by mixing 2.5 μl (12.5 ng) of genomic DNA extracted from the control and experimental groups isolated above with 1 μm of each primer and 2x KAPA Hifi Hotstart Ready Mix (KAPA Bio, UK) premixture to make 25 μl. The PCR reaction was performed using an Agilent 2200 TapeStation (Agilent Tech, Italy) with denaturation at 95°C for 5 minutes, followed by 27 cycles of 95°C for 30 seconds, 55°C for 30 seconds, and 72°C for 30 seconds, and finally an extension step at 72°C for 5 minutes. The obtained PCR products were classified into V1-V2 (340-370 bp) and V3-V4 (440-460 bp), purified using the Agencourt AMPure XP kit (Beckman, Iatly), and a library was completed. The nucleotide sequences were analyzed using Illumina MiSeq against 19 strains notified by the Ministry of Food and Drug Safety (October 2020; Appendix 1-2~4) https: / / www.foodsafetykorea.go.kr / foodcode / 01_03.jsp?idx=12135) and non-notified strains Lactobacillus brevis and Leuconostoc mesentroides added to feed. The results are shown in Table 8 and Figure 3.

[0153] number strain name Strain distribution (%) in the control group Strain distribution (%) within the experimental group 1 Lactobacillus acidophilus - - 2 Lactobacillus casei - - 3 Lactobacillus gasseri - - 4 Lactobacillus delbrueckii - - 5 Lactobacillus helveticus - - 6 Lactobacillus fermentum - - 7 Lactobacillus paracasei - - 8 Lactobacillus plantarum - 0.6 9 Lactobacillus reuteri - - 10 Lactobacillus rhamnosus - - 11 Lactobacillus salivarius - - 12 Lactococcus lactis - 61.8 13 Enterococcus faecium - 32.1 14 Enterococcus faecalis - - 15 Streptococcus thermophilus - - 16 Bifidobacterium bifidum - - 17 Bifidobacterium breve - - 18 Bifidobacterium longum - - 19 Bifidobacterium animalis - - 20 Lactobacillus brevis 0.9 21 Leuconostoc mesentroides 4.6 total - 0 100

[0154] As shown in Table 8 above, in the control group of cherry salmon fed with the control feed for 4 weeks, none of the 21 strains of the control were identified in the intestinal microorganisms.

[0155] In contrast, it was confirmed that a total of 5 types of intestinal microorganisms were distributed in the intestines of the experimental group of mountain trout fed with the experimental samples for 4 weeks, consisting of 3 types of 19 types of lactic acid bacteria and 2 types of non-notified bacteria. Specifically, Lactococcus lactis, a lactic acid bacterium administered to HiLAB, accounted for the majority at 61.8%, and Enterococcus faecium accounted for 32.1%. The remaining 3 types were confirmed to be present as Lactobacillus plantarum (0.6%), Leuconostoc mesentroides (4.6%), and Lactobacillus brevis (0.9%), which were administered as notified bacteria.

[0157] Example 6. Confirmation of changes in fatty acids in cherry salmon due to consumption of fermented black soldier fly extract

[0158] Example 6-1. Measurement of unsaturated fatty acids in cherry salmon

[0159] The unsaturated fatty acid content in the control group and experimental group trout fed the feed in Example 4-2 above was measured.

[0160] Specifically, the flesh of the trout, with the internal organs removed, was collected and treated with a methanolic sodium hydroxide solution. Subsequently, a trifluoroborane methanol solution was added, and the mixture was heated to induce esterification. The resulting fatty acid esters were dissolved in isooctane and analyzed using gas chromatography (US / HP 6890, Aglient Technologies, Youido, Korea). Undecanoic acid was used as the standard solution. For the gas chromatography (GC) conditions, a silica capillary column (Omegawax 205, 0.25 μm film thickness) was used, with the injection port temperature set to 225°C and the detector temperature maintained at 285°C. The column temperature was raised from 100°C to 250°C and maintained for at least 15 minutes. The above process was repeated twice to analyze the fatty acid content, and the results are shown in Table 9 below. The measured content values ​​are expressed as mean ± standard deviation. The unit of the content values ​​measured below is μg / g muscle weight, and the unit of all content values ​​measured in Example 6 is μg / g muscle weight.

[0161] Free fatty acids O.masou control group Experimental group (HiLAB) C14 Myristic acid 2.47±0.01 2.52±0.10 C14:1 myristoleic acid 0.37±0.01 0.45±0.01 C16 Palmitic acid 18.88±0.44 15.18±0.36 C16:1 Palmitoleic acid 0.63±0.09 0.98±0.09 C18 Stearic acid 6.18±0.15 4.99±0.12 C18:1 Oleic acid 20.76±0.47 22.31±0.49 C18:2 Linoleic acid 2.91±0.07 3.14±0.07 C18:3 α-Linolenic acid 1.48±0.03 1.59±0.03 C20:0 Eicosanoic acid 0.31±0.00 0.39±0.01 C20:1 Gadoleic acid 0.34±0.01 0.28±0.01 C20:3 Eicosatrienoic acid 0.02±0.00 0.03±0.00 C20:4 Arachidonic acid 0.12±0.00 0.12±00 C20:5 Eicosapentaenoic acid 0.38±0.01 0.44±0.01 C22:0 Behenic acid 0.57±0.01 0.56±0.01 C22:1 Erucic acid 0.30±0.01 0.34±0.01 C22:6 Docosahexaenoic acid 0.56±0.00 0.82±0.01 C24:0 Lignoceric acid 0.08±0.00 0.09±0.00 C24:1 Nervonic acid 0.16±0.00 0.14±0.00

[0162] As shown in Table 9 above, both the control group and the experimental group were measured to have high levels of palmitic acid and oleic acid, with palmitic acid being measured to be higher in the control group and oleic acid being measured to be higher in the experimental group.

[0163] Example 6-2. Measurement of saturated and unsaturated fatty acid content in cherry salmon

[0164] Based on the fatty acid content measured in Example 6-1 above and the number of carbon chains presented in Table 9, the content of saturated fatty acids (fatty acids having one or more single bonds in the fatty acid chain; saturated fatty acid; SFA) and unsaturated fatty acids (fatty acids having one or more double bonds in the fatty acid chain; Unsaturated fatty acid; USFA) in the trout was measured separately, and the content of monounsaturated fatty acids (cases containing one double bond; monounsaturated fatty acids; MUFAs) and polyunsaturated fatty acids (fatty acids having two or more double bonds in the fatty acid chain; polyunsaturated fatty acid; PUFA) among the unsaturated fatty acids in the trout was also measured, and the results are shown in Figures 4 and 5, respectively.

[0165] As shown in Figure 4, the content of saturated fatty acids (SFA) was measured to be higher in the control group, and the content of unsaturated fatty acids (UFA) was measured to be higher in the experimental group, confirming that the content of unsaturated fatty acids was higher in the trout that consumed feed containing HiLAB.

[0166] In addition, as shown in Figure 5, the content of both monounsaturated fatty acids and polyunsaturated fatty acids was measured to be high in the experimental group, confirming that the content of both types of unsaturated fatty acids was higher in trout that consumed feed containing HiLAB.

[0167] Example 6-3. Measurement of Omega-3, Omega-6, and Omega-9 Unsaturated Fatty Acid Content in Cherry Salmon

[0168] The content of omega-3, omega-6, and omega-9 related unsaturated fatty acids in cherry salmon was measured in substantially the same way as in Example 6-1 above, and the results are shown in Figure 6.

[0169] As shown in Figure 6, the content of the anti-inflammatory omega-3 unsaturated fatty acids α-linolenic acid (ALA) (C18:3), eicosapentaenoic acid (EPA) (C20:5), and docosahexaenoic acid (DHA) (C22:6) was all measured to be high in the experimental group, the content of the omega-6 unsaturated fatty acids α-linolenic acid (C18:2) and eicosapentaenoic acid (C20:3) was measured to be high in the experimental group, the content of arachidonic acid (C20:4) was measured to be similar in the control and experimental groups, and the content of the anti-inflammatory omega-9 unsaturated fatty acids oleic acid (C18:1) and erucic acid (C22:1) was also all measured to be high in the experimental group. Based on the above measurement results, it was confirmed that the content of omega-3, omega-6, and omega-9 unsaturated fatty acids was higher in cherry salmon fed a diet containing HiLAB, or was equivalent to or greater than that in cherry salmon fed a diet not containing HiLAB.

[0171] Name of depositing institution: Korean Culture Collection of Microorganisms Trustee Number: KCCM13066P Date of Deposit: 2021-10-27 Name of depositing institution: Korean Culture Collection of Microorganisms Trustee Number: KCCM13067P Date of Deposit: 2021-10-27 Name of depositing institution: Korean Culture Collection of Microorganisms Trustee Number: KCCM13068P Date of Deposit: 2021-10-27 Name of depositing institution: Korean Culture Collection of Microorganisms Trustee Number: KCCM13069P Date of Deposit: 2021-10-27

Claims

Claim 1 It comprises a lactic acid fermented product of a black soldier fly solvent extract, wherein the lactic acid bacteria are Lactobacillus plantarum, Lactococcus lactis, Leuconostoc mesenteroides, and Lactobacillus brevis, wherein the Lactobacillus plantarum is the MIRE_TS55 strain deposited under accession number KCCM13068P, the Lactococcus lactis is the MIRE-P1 strain deposited under accession number KCCM13066P, the Leuconostoc mesenteroides is the MIRE_TS1 strain deposited under accession number KCCM13067P, and the Lactobacillus brevis is the MIRE_TS66 strain deposited under accession number KCCM13069P. A feed composition characterized by being a strain. Claim 2 A feed composition according to claim 1, wherein the black soldier fly solvent extract is a solvent extract obtained by extracting black soldier flies with water, a straight-chain or branched-chain alcohol having 1 to 4 carbon atoms, or a mixture thereof. Claim 3 A feed composition according to paragraph 2, wherein the black soldier fly extract is a solvent extract of black soldier fly larva powder. Claim 4 A feed composition according to claim 1, wherein the lactic acid bacteria fermented product is obtained by adding one or more carbon sources selected from the group consisting of dextrose, galactose, mannose, sucrose, glucose, starch, fructose, lactose, maltose, cellulose, lactulose, trehalose, cellobiose, chitobiose, kozibiose, nigerose, isomaltose, sophorose, laminaribiose, gentiobiose, turanos, maltulose, isomaltulose, gentiobiulose, mannovios, melivios, beribiulose, rutinose, rutinulose, xylobiose, and rhamnose to a black soldier fly solvent extract and culturing lactic acid bacteria. Claim 5 A feed composition according to claim 4, wherein the lactic acid bacteria fermented product is a black soldier fly solvent extract to which lactic acid bacteria are cultured by additionally adding one or more nitrogen sources selected from the group consisting of yeast, milk casein, peptone, soy peptone, casein peptone, meat peptone, polypeptone, tryptone, soytone, and beef extract, along with one or more carbon sources. Claim 6 delete Claim 7 A feed composition according to claim 1, wherein the feed is intended for use in fish, shrimp, chickens, or pets. Claim 8 A feed composition according to claim 7, wherein the fish is one or more selected from the group consisting of cherry salmon, spotted flounder, stone flounder, halibut, rockfish, rainbow trout, yellowtail, eel, carp, ornamental fish, catfish, herring, sea bass, cod, anglerfish, pufferfish, scorpionfish, horse mackerel, skate, and sweetfish. Claim 9 A feed composition according to claim 7, wherein the pet is a dog or a cat. Claim 10 A feed composition in which, in claim 7, the chicken is a broiler or a laying hen. Claim 11 A feed composition according to claim 1, wherein the content of the lactic acid bacteria fermented product of the black soldier fly solvent extract is 1 to 10 weight%. Claim 12 A feed composition according to claim 1, wherein the lactic acid fermented product of the black soldier fly solvent extract has one or more characteristics selected from the group consisting of the following (1) to (3): (1) growth-promoting ability of animals; (2) increase in the ratio of beneficial bacteria in the intestines; and (3) improvement of lipid metabolism in animals. Claim 13 A feed composition according to claim 12, wherein the improvement of lipid metabolism in the animal body means an increase in the content of unsaturated fatty acids in the animal body. Claim 14 In claim 12, the feed composition is characterized in that, when the above feed composition is consumed, the daily growth rate calculated by the method of [Mathematical Formula 2] below, measured at the 4th week after consumption of the above feed composition, is 1.01 to 3 times the daily growth rate calculated by the method of [Mathematical Formula 2] below, measured at the 4th week after consumption of the feed not containing the lactic acid bacteria fermented product of the black soldier fly solvent extract, when the above feed is consumed: [Mathematical Formula 2] . Claim 15 A feed composition according to claim 12, wherein the beneficial intestinal bacteria are one or more selected from the group consisting of Lactococcus lactis, Enterococcus faecium, Lactobacillus plantarum, Leuconostoc mesentroides, and Lactobacillus brevis. Claim 16 A feed composition according to claim 13, wherein the unsaturated fatty acid is one or more selected from the group consisting of myristoleic acid, palmitoleic acid, oleic acid, linoleic acid, α-linolenic acid, gadoleic acid, eicosatrienoic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, and nervonic acid. Claim 17 A feed composition according to claim 13, wherein the unsaturated fatty acid is one or more selected from the group consisting of omega-3 unsaturated fatty acids, omega-6 unsaturated fatty acids, and omega-9 unsaturated fatty acids. Claim 18 A feed composition that is a composition for improving lipid metabolism in an animal body, in any one of claims 1 to 5 and 7 to 17. Claim 19 delete Claim 20 delete Claim 21 delete Claim 22 delete

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