Composition for rearing ostriches

Abstract

Provided is a composition for rearing ostriches, the composition comprising: cells of a bacterium belonging to Lactiplantibacillus plantarum; a bacterial cell culture; or extracts thereof.

Description

Composition for breeding ostriches

[0001] The present invention relates to a composition for breeding ostriches.

[0002] With the global population increase and economic development, the shortage of protein supply, one of the three major nutrients, has become serious. Ostriches can obtain the same amount of edible meat with less feed (e.g., grains) compared to cows. Therefore, breeding ostriches to produce edible meat has been attracting attention as a new protein source. Since the breeding method of ostriches is still in the development stage, various breeding methods are being studied.

[0003] For example, Japanese Patent Application Laid-Open No. 2011218616 (Patent Document 1) discloses an auxiliary feed composition for ostriches containing at least one of Lactobacillus oris, Lactobacillus brevis, Lactobacillus johnsonii, Bifidobacterium pseudolongum subsp. globosum, and Enterococcus faecalis.

[0004] Japanese Patent Application Laid-Open No. 2011218616

[0005] The survival rate of ostriches is easily affected by the environment such as the temperature and humidity of the land. For example, breeding in a hot and humid region like Japan results in a lower growth rate (e.g., weight gain during growth) and a lower survival rate of chicks compared to other regions (e.g., regions with a savannah climate). Therefore, improvement of feed and breeding methods is desired.

[0006] The present invention has been made in view of the above circumstances, and an object thereof is to provide a breeding composition capable of increasing the weight or improving the survival rate of ostriches.

[0007] As a result of intensive research, the present inventors have found that by allowing bacteria belonging to the species Lactiplantibacillus plantarum, their cell bodies, cell cultures, or extracts thereof to be ingested by ostriches, it is possible to increase the weight or improve the survival rate, and thus completed the present invention. That is, the present invention relates to the items exemplified below.

[0008] [1] A composition for ostrich rearing comprising bacterial cells or bacterial cultures of a bacterium belonging to the species Lactiplantibacillus plantarum, or extracts thereof. [2] The composition for ostrich rearing according to [1], wherein the bacterial cells are heat-sterilized. [3] The composition for ostrich rearing according to [1] or [2], wherein the composition for ostrich rearing comprises bacterial cells, and the content of the bacterial cells is 0.0001 to 0.01% by mass relative to the composition for ostrich rearing. [4] The composition for ostrich rearing according to any one of [1] to [3], further comprising basic feed or drinking water. [5] The composition for ostrich rearing according to [4], further comprising the basic feed, wherein the basic feed comprises protein or lipid. [6] The composition for ostrich rearing according to any one of [1] to [5], further comprising at least one selected from the group consisting of dextrin, vitamins, trace inorganic salts, organic acids, and enzymes. [7] A method for raising ostriches, comprising the step of having ostriches ingest an ostrich breeding composition described in any of [1] to [6]. [8] The method for raising ostriches according to [7], wherein the ostrich includes a chick. [9] The method for raising ostriches according to [7] or [8], wherein the ostrich includes a North African ostrich, a Masai ostrich, a South African ostrich, a Somali ostrich, a Mauritanian ostrich, or an African black ostrich.

[10] A method for producing ostrich-derived food ingredients, comprising the steps of raising ostriches by the method for raising ostriches according to any of [7] to [9], and processing the raised ostriches into food ingredients.

[11] A bacterium having a 16S rRNA gene containing a nucleotide sequence having 99.8% or more sequence identity with the nucleotide sequence described in Sequence ID No. 1, and having a weight-increasing effect or a survival rate-improving effect on ostriches.

[12] The bacterium according to

[11] , having a 16S rRNA gene containing the nucleotide sequence described in Sequence ID No. 1.

[13] Bacteria deposited under accession number NITE-BP-04252.

[0009] According to the present invention, it is possible to provide a breeding composition that can increase the weight of ostriches or improve their survival rate.

[0010] This figure shows (A) the shape of the colony and (B) the Gram staining results for NITE-BP-04252 in Experiment 1. This graph shows the effect of NITE-BP-04252 on weight gain in ostrich chicks in Experiment 2. The vertical axis shows the amount of weight gain. The horizontal axis shows the age in days. This graph shows the effect of NITE-BP-04252 on improving the survival rate of ostrich chicks in Experiment 2. The vertical axis shows the survival rate. The horizontal axis shows the age in days.

[0011] Hereinafter, one embodiment of the present invention (hereinafter referred to as "this embodiment") will be described. However, this embodiment is not limited thereto. In this specification, the notation in the form of "A to Z" means the upper and lower limits of the range (i.e., A or more and Z or less), and if no unit is specified for A and only a unit is specified for Z, the unit for A and the unit for Z are the same.

[0012] <Composition for Ostrich Rearing> The composition for ostrich rearing according to this embodiment contains bacterial cells or bacterial cultures of bacteria belonging to the species Lactiplantibacillus plantarum, or extracts thereof.

[0013] In this embodiment, "ostrich feeding composition" means a composition intended for ingestion by ostriches. The ostrich feeding composition includes feed or drinking water or additives to be added thereto. The ostrich feeding composition may be solid or liquid.

[0014] (Bacterial cells of the species Lactiplantibacillus plantarum) In this embodiment, "bacteria belonging to the species Lactiplantibacillus plantarum" (hereinafter sometimes referred to as "L. plantarum") refers to a rod-shaped bacterium belonging to the phylum Bacillota, class Bacilli, order Lactobacillales, family Lactobacillaceae, and is a Gram-positive rod-shaped bacterium. The above L. plantarum is not particularly limited, and examples include NBRC15891, NBRC3070, and NBRC3074. In this embodiment, the above L. plantarum is preferably NITE-BP-04252. NITE-BP-04252 is considered highly safe when used in ostrich rearing because it exists in the natural environment. NITE-BP-04252 may also be an isolated bacterium.

[0015] Conventionally, it has been known that when ostriches are raised in hot and humid regions such as Japan, their growth rate (e.g., weight gain during growth) is lower and the survival rate of chicks is lower compared to other regions (e.g., regions with a savanna climate). As a result of diligent research, the inventors have found that by having ostriches ingest L. plantarum such as NITE-BP-04252, it is possible to increase the weight of the ostriches or improve their survival rate. In other words, the above ostrich rearing composition, when ingested by ostriches, makes it possible to increase the weight of the ostriches or improve their survival rate. In particular, since it is possible to significantly improve the survival rate even in hot and humid regions such as Japan, the above ostrich rearing composition is suitable for raising ostriches in hot and humid regions. In this embodiment, "hot and humid region" means a region where the average temperature is 30°C or higher and 36°C or lower and the average humidity is 55% or higher and 77% or lower.

[0016] NITE-BP-04252 is a bacterium that has been internationally deposited with the National Institute of Technology and Evaluation (NPMD, address: Room 122, 2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture 292-0818, Japan) under the Budapest Convention, with accession number NITE-BP-04252 (original deposit date: December 26, 2024). NITE-BP-04252 is a bacterium belonging to the species Lactiplantibacillus plantarum, a type of lactic acid bacterium. The mycological properties of the above bacterium are shown in Tables 1 and 2 and Figure 1 below.

[0017] The ostrich rearing composition according to this embodiment contains L. plantarum cells or cell cultures or extracts thereof. The cells may be cells isolated from the environment or cultured cells. The cells may be live cells or heat-sterilized cells. The cells may be present in a culture medium, buffer solution, etc., or in a concentrated state from which the liquid component has been removed. The cells may also be present in a dried state of the culture medium (e.g., freeze-dried product) or in a frozen stock state. Any known method may be used to heat-sterilize the cells. For example, a method of heat-sterilizing the cells at 70°C for 1 hour may be used.

[0018] The bacterial culture comprises bacteria (L. plantarum) and a culture medium. The culture medium may be a liquid medium or a solid medium. The bacterial culture may contain bacterial secretions, metabolites, etc. The bacterial culture may also contain peptides, proteins, sugars, enzymes, or organic acids produced by the bacteria.

[0019] The above-mentioned L. plantarum can be cultured according to the usual methods for culturing lactic acid bacteria. Typical culture methods include culturing at a temperature of 30°C using MRS (de Man, Rogosa and Sharp) liquid medium or MRS agar medium.

[0020] Extracts of bacterial cells or bacterial cultures are prepared in such a way that they do not lose the weight-gaining effect or viability-enhancing effect of the L. plantarum bacterial cells or bacterial cultures. The extracts can be obtained, for example, by treating bacterial cells or bacterial cultures with ultrasonic disruption, bead grinding, freeze-thaw cycles, chemical dissolution, etc. The extracts may also be obtained by salting out, ultrafiltration, ion-exchange chromatography, or liquid-phase extraction using organic solvents. These treatments can be carried out in appropriate combinations. The extracts may contain bacterial cell fragments, nucleic acids, peptides, proteins, sugars, and enzymes. In this specification, bacterial cells or bacterial cultures or extracts thereof are also referred to as "bacterial preparations."

[0021] If the above ostrich breeding composition contains bacterial cells, the bacterial cell content may be 0.0001 to 0.01% by mass, 0.00001 to 0.001% by mass, or 0.001 to 0.1% by mass relative to the ostrich breeding composition. The amount of bacterial cells can be quantified by fluorescence in situ hybridization (FISH method). The base sequence of the oligonucleotide used as a probe is as follows. Needless to say, the FISH method can also be used to qualitatively measure the presence or absence of the above bacterial cells in the ostrich breeding composition. Probe name: Lpb V3 Base sequence: CTGTTCAGGTTATTGACG (SEQ ID NO: 2)

[0022] (Other components) The above ostrich rearing composition may further contain basic feed or drinking water. In one aspect of this embodiment, the above ostrich rearing composition may further contain at least one selected from the group consisting of dextrin, vitamins (e.g., vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin D, vitamin E, calcium pantothenate, nicotinamide, folic acid, etc.), trace inorganic salts (e.g., magnesium sulfate, iron sulfate, copper sulfate, zinc sulfate, potassium iodide, cobalt sulfate, etc.), organic acids (e.g., formic acid, propionic acid, etc.), and enzymes (e.g., phytase, etc.).

[0023] The above-mentioned basic feed may contain protein or lipids.

[0024] The basal feed used in the ostrich feed composition of this embodiment is not particularly limited as long as it is a basal feed used for raising poultry, and may be manufactured by known methods or purchased as a commercially available product. Examples of commercially available products include the products described in the examples below. Examples of raw materials used in the manufacture of the basal feed include corn, rice, wheat, milo, soybean meal, rapeseed meal, bran, defatted rice bran, fish meal, skim milk powder, dried whey, alfalfa meal, North Pacific meal, soybean oil, powdered refined beef tallow, wheat flour, rapeseed oil, meat and bone meal (feather meal), animal fats and oils, calcium phosphate, corn gluten meal, corn distiller's grain soluble, molasses, calcium carbonate, tricalcium phosphate, and sodium chloride. Examples include choline chloride, vitamins (vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin D, vitamin E, calcium pantothenate, nicotinamide, folic acid, etc.), amino acids (lysine, methionine, threonine, etc.), trace inorganic salts (magnesium sulfate, ferrous sulfate, copper sulfate, zinc sulfate, potassium iodide, cobalt sulfate, etc.), organic acids (formic acid, propionic acid, etc.), enzymes (phytase, etc.), and probiotics.

[0025] The above-mentioned ostrich rearing composition may contain bacteria other than L. plantarum. Examples include lactic acid bacteria, bifidobacteria, yeast, Bacillus subtilis, Aspergillus oryzae, butyric acid bacteria, acetic acid bacteria, and the like.

[0026] The above-mentioned ostrich rearing composition can be manufactured by known methods. For example, the following manufacturing method can be used. First, the feed pellets, which are the basic feed, are weighed. The weighed feed pellets are coarsely ground in a mixer to about half their original size. A portion of the coarsely ground feed pellets and the L. plantarum fungal cells are added to a plastic bag and thoroughly mixed in the bag. Then, the remaining portion of the coarsely ground feed pellets is added to the plastic bag and mixed further to obtain the ostrich rearing composition. The weighing at this stage can be done using a balance or the like.

[0027] In one aspect of this embodiment, the proportion of L. plantarum fungal cells added during the production of the ostrich breeding composition may be 0.0001 to 0.01% by mass, 0.00001 to 0.001% by mass, or 0.001 to 0.1% by mass, relative to the ostrich breeding composition produced.

[0028] In one aspect of this embodiment, the proportion of other raw materials used in the production of the ostrich breeding composition can be appropriately set within the range in which the effects of the present invention are achieved.

[0029] <Weight-increasing agent, survival rate-enhancing agent> The ostrich feed composition according to this embodiment can increase the weight or improve the survival rate of ostriches when ingested. Therefore, the above ostrich feed composition can also be understood as a weight-increasing agent for ostriches or a survival rate-enhancing agent for ostriches. Specifically, the weight-increasing agent for ostriches according to this embodiment contains bacterial cells or bacterial cultures of bacteria belonging to the species Lactiplantibacillus plantarum, or extracts thereof. The survival rate-enhancing agent for ostriches according to this embodiment contains bacterial cells or bacterial cultures of bacteria belonging to the species Lactiplantibacillus plantarum, or extracts thereof.

[0030] <Ostrich Rearing Method> The ostrich rearing method according to this embodiment includes the step of having the ostrich ingest the above-mentioned ostrich rearing composition. The specific form of the above-mentioned ostrich rearing composition is as described above.

[0031] The method for administering the above-mentioned ostrich feeding composition to ostriches is not particularly limited, and any known method can be used. For example, the ostrich feeding composition may be freely administered to ostriches, or it may be administered automatically at regular intervals using an automatic feeder in which the ostrich feeding composition is stored. If the ostrich is a chick, the ostrich feeding composition may be administered to the chick by hand feeding. The above-mentioned ostrich feeding composition may be in liquid or solid form. The method for administering the above-mentioned ostrich feeding composition to ostriches may be appropriately modified depending on the growth stage of the ostrich.

[0032] The above ostrich rearing composition may be administered once or multiple times per day. The above rearing composition may also be administered once every few days. The period for administering the above rearing composition can be appropriately selected depending on the type of ostrich, etc. The above rearing composition containing L. plantarum cells may be administered continuously for the entire rearing period, or for only a portion of the period. "A portion of the period" may be, for example, 10% or more, 20% or more, 30% or more, 50% or more, 70% or more, or 90% or more of the entire rearing period. In one aspect of this embodiment, the upper limit of the above-mentioned "a portion of the period" is not particularly limited, but may be, for example, less than 100% of the entire rearing period, or 99% or less. The period for administering the above rearing composition containing L. plantarum cells may be, for example, 2 weeks to 17 months (for example, 14 days to 489 days), or about 2 months (for example, 59 days to 61 days). The above-mentioned L. plantarum fungal cell-containing rearing composition may be administered repeatedly with interruptions at any time.

[0033] The ostriches mentioned above may include chicks. In this embodiment, chicks refer to ostriches that are 0 to 3 months old (for example, 0 to 92 days old).

[0034] The ostriches mentioned above may include North African ostriches, Masai ostriches, South African ostriches, Somali ostriches, Mauritanian ostriches, or African black ostriches. In particular, African black ostriches may be included.

[0035] In this embodiment, the period for raising ostriches in this process is not particularly limited, but may be, for example, 1 month to 17 months (e.g., 28 days to 489 days), or 2 months to 12 months (e.g., 28 days to 366 days).

[0036] <Method for producing ostrich-derived food ingredients> The method for producing ostrich-derived food ingredients according to this embodiment includes the steps of raising ostriches using the above-described method of raising ostriches, and processing the raised ostriches into food ingredients.

[0037] (Process of raising ostriches) The process of raising ostriches described above is carried out according to the ostrich raising method described above. The specific implementation is the same as the ostrich raising method described above.

[0038] (Process for processing ostrich into food ingredients) In this embodiment, "processing" means adding something to a whole. The above "processing" does not specify the degree of adding something (degree of processing). Examples of "processing" in this embodiment include "poultry processing" as defined in Article 2, Item 5 of the "Act on Regulation of Poultry Processing Businesses and Inspection of Poultry (Act No. 70 of 1990)" in Japan.

[0039] In this embodiment, "ostrich-derived ingredients" means ingredients obtained by processing ostriches. The ostrich-derived ingredients may include edible meat, skin, bones, or organs, or extracts obtained from these. Examples of the extracts include dashi (Japanese soup stock). The ostrich-derived ingredients may also include edible meat, skin, or organs that have been seasoned with seasonings. The edible meat may or may not have bones.

[0040] The above method for producing the ostrich-derived food ingredient may further include a step of seasoning the food ingredient with seasonings or a step of packaging the processed food ingredient.

[0041] <Bacteria with weight-increasing or survival-enhancing effects on ostriches> The bacteria according to this embodiment have a weight-increasing or survival-enhancing effect on ostriches. These bacteria have a 16S rRNA gene containing a nucleotide sequence that has 99.8% or more sequence identity with the nucleotide sequence described in Sequence ID No. 1, and thus have a weight-increasing or survival-enhancing effect on ostriches.

[0042] In the present embodiment, "sequence identity" means the percentage (%) of identical bases with respect to all overlapping base sequences in an optimal alignment (preferably, the algorithm can consider the introduction of gaps into one or both of the sequences for optimal alignment) when two base sequences are aligned using a mathematical algorithm known in the art. The "sequence identity" of a base sequence can be easily confirmed by those skilled in the art. For example, NCBI BLAST (National Center for Biotechnology Information Basic Local Alignment Search Tool) can be used.

[0043] In one aspect of the present embodiment, the bacterium may have a 16S rRNA gene containing the base sequence set forth in SEQ ID NO: 1, or may have a 16S rRNA gene consisting of the base sequence set forth in SEQ ID NO: 1. Further, the bacterium may be a bacterium deposited under accession number NITE-BP-04252.

[0044] In the present embodiment, the "weight gain effect" is determined to exist when the weight gain of the ostrich at 35 days of age is greater than the weight gain of the control group at the same time point. "Weight gain" means the weight increased during the period from the start of breeding to the time of weight measurement. The weight gain is obtained by the following formula 1. Here, the "control group" means a group of ostriches that have not ingested the bacterium. The "start point of breeding" means the start point of breeding in the chick house (see Examples). (Weight gain) g = (weight at the measurement time point) - (weight at the start point of breeding) (Formula 1)

[0045] In the present embodiment, the "survival rate improvement effect" is determined to exist when the survival rate of the ostrich at 35 days of age is greater than the survival rate of the control group at the same time point. The survival rate is obtained by the following formula 2. (Survival rate) % = 100 × (number of individuals surviving at the measurement time point) / (number of individuals surviving at the start point of breeding) (Formula 2)

[0046] Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

[0047] [Experiment 1: Isolation and Identification of NITE-BP-04252] The separation source (Quercus acutissima) was ground together with sterilized water. The ground liquid was appropriately diluted and added to a 1 / 2 MRS liquid medium for enrichment culture. The enrichment culture solution was spread on an MRS agar medium containing calcium carbonate, and microorganisms forming halos were isolated. Hereinafter, this isolate is referred to as isolate A. When the culture solution of isolate A was suspended in hydrogen peroxide, no bubbles were generated. Since isolate A has no catalase activity, it was confirmed to be a lactic acid bacterium.

[0048] Isolate A was identified by 16S rRNA gene analysis, morphological observation, and physiological and biochemical property tests. (1) 16S rRNA gene analysis Genomic DNA was extracted from isolate A, and using the obtained genomic DNA as a template, PCR amplification of the 16S rRNA gene was performed using a cloning forward primer 9F and a cloning reverse primer 1510R (Yasuyoshi Nakagawa et al.: Gene Analysis Method, Base Sequence Determination Method of 16S rRNA Gene, edited by the Japanese Society for Actinomycetes, Classification and Identification of Actinomycetes, 88-117 pp., Japan Society Affairs Center, 2001). PCR amplification was performed using Tks Gflex DNA polymerase (manufactured by Takara Bio Inc.), and the amplified product after PCR was purified.

[0049] A cycle sequencing reaction was performed using the purified amplified product after PCR. The cycle sequencing reaction was performed using the BigDye Terminator v3.1 Cycle Sequencing Kit. The obtained reaction solution was purified, and the purified solution was subjected to DNA sequence analysis (3130xl DNA Analyzer) to determine the base sequence of the 16S rRNA gene of the template DNA extracted from isolate A. As primers for sequence analysis, 9F, 515F, 1099F, 536R, 926R, 1510R (Yasuyoshi Nakagawa et al.: Gene Analysis Method, Base Sequence Determination Method of 16S rRNA Gene, edited by the Japanese Society for Actinomycetes, Classification and Identification of Actinomycetes, 88-117 pp., Japan Society Affairs Center, 2001) were used.

[0050] The nucleotide sequence of the 16S rRNA gene of isolate A was analyzed using the microbial identification system "ENKI" (Techno Suruga Labs) and performed a BLAST homology search against the microbial identification database DB-BA15.0 (Techno Suruga Labs) and the international nucleotide sequence database (DDBJ / ENA(EMBL) / GenBank). The nucleotide sequence of the 16S rRNA gene of isolate A showed 99.9% identity with the 16S rRNA gene of Lactobacillus pentosus (JCM1558) in both the microbial identification database and the international nucleotide sequence database, indicating Lactobacillus plantarum subsp. The nucleotide sequence of the 16S rRNA gene of plantarum (JCM1149) showed 99.8% identity in the microbial identification database and 99.9% identity in the international nucleotide sequence database. However, no microorganisms were found that possessed a 16S rRNA gene that perfectly matched the nucleotide sequence of isolate A.

[0051] (2) Morphological observation and physiological / biochemical characterization tests Isolated bacteria A was spread on MRS agar medium and cultured aerobically at 30°C for 72 hours. Cell morphology, Gram staining, motility, and colony morphology were observed using the following methods. Colony morphology was observed using a stereomicroscope SMZ800N (Nikon Corporation). Cell morphology was observed using an optical microscope BX50F4 (Olympus Corporation). For Gram staining, Faber G "Nissui" (Nissui Pharmaceutical Co., Ltd.) was used. Based on the method described in Barrow & Feltham (Cowan and Steel's Manual for the Identification of Medical Bacteria, 3rd ed. Cambridge: Cambridge University Press; 1993), tests were conducted on catalase reaction, oxidase reaction, acid / gas production from glucose, and oxidation / fermentation (O / F) of glucose. The physiological and biochemical properties of bacteria were investigated using the API50CHB kit (bioMerieux, France).

[0052] As shown in Figure 1(A), isolate A formed circular colonies. As shown in Figure 1(B), isolate A was Gram-positive. The results of the physiological and biochemical characterization tests and fermentation tests of isolate A are shown in Tables 1 and 2. Isolate A was a non-motile, Gram-positive rod that did not form spores. Isolate A was negative for catalase and oxidase reactions and fermented glucose. These characteristics were consistent with those of the genus Lactiplantibacillus, to which 16S rDNA partial sequencing analysis suggested a possible affinacy. In fermentation tests using an API kit, isolate A fermented galactose, fructose, etc., but did not ferment glycerol, D-xylose, etc. Isolate A did not show arginine dihydrolase activity and grew at 10°C. These characteristics were consistent with those of L., to which 16S rDNA partial sequencing analysis suggested a possible affinacy. Of L. pentosus and L. plantarum, isolate A differed from L. pentosus in that it did not exhibit glycerol and D-xylose fermentation, and its characteristics were consistent with L. plantarum. Therefore, isolate A was found to be a novel isolate belonging to Lactibactibacillus plantarum (a bacterium belonging to the Lactibactibacillus plantarum species). Isolate A was internationally deposited as NITE-BP-04252. Isolate A was stored at -80°C as a frozen stock using a known method until Experiment 2, described later, was performed.

[0053]

[0054]

[0055] [Experiment 2: Verification of the effect of L. plantarum ingestion on weight gain or improved survival rate] This experiment investigated whether orally ingested L. plantarum cells had an effect on weight gain or improved survival rate in ostrich chicks. The experiment was conducted from mid-July to early September 2023. The average daytime temperature outdoors during the experiment period was 30°C to 34°C, and the average humidity was 51% to 74%. Twelve ostriches (7 males and 5 females) were used in each test group. The specific procedure is described below.

[0056] (1: Production of feed additive containing isolate A) Isolate A, which had been stored as frozen stock, was awakened using a known method, then transferred to MSR liquid medium and cultured with shaking at 30°C for 24 hours. At this time, a plastic bag manufactured by ZACROS (product name: Culture Bag CB20-1) was used as the culture container.

[0057] The plastic bags containing the cultured cells were transferred to an autoclave and sterilized by heat (70°C for 10 minutes) using the dissolution and heat retention function. The bacterial cultures in the plastic bags were then transferred to a centrifuge tube (himac Co., Ltd., product name: 1000PP bottle (WN)) and centrifuged using a himac Co., Ltd. centrifuge (product name: CR22N) at 4°C, 8000 × g, for 5 minutes. The supernatant was removed from the centrifuge tube, and 20 times the volume of the bacterial cells was added to the tube. The centrifuge tube was washed by manually shaking it to suspend the bacterial cells. The centrifuge tube containing the bacterial suspension was then subjected to centrifugation and washing with ion-exchanged water using the same method as described above. This series of operations was repeated a total of three times.

[0058] Subsequently, the centrifuge tube containing the bacterial suspension was centrifuged using the same method as described above to recover only the bacterial cells. The recovered bacterial cells were freeze-dried at -60°C for 72 hours using a freeze-dryer manufactured by CHRISTO (product name: Alpha 1-2 LDplus) to obtain dried bacterial cells (heat-sterilized bacterial cells) of isolate A.

[0059] The above-mentioned dried bacterial cells and dextrin powder (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., product name: dextrin hydrate) were added to a mortar in a mass ratio of 1:9, and then crushed and mixed with a pestle. Following this procedure, a feed additive containing heat-sterilized bacterial cells of NITE-BP-04252 was obtained. This feed additive can be understood as one embodiment of the ostrich rearing composition of the present invention.

[0060] (2: Production of feed containing isolate A) First, 5 kg of feed pellets (approximately φ3 mm, approximately 7 mm in length) (manufactured by Jonan Green System Co., Ltd., product name: Starter Feed 20 KG), which are the base feed, were weighed. The weighed feed pellets were coarsely ground in a mixer (manufactured by TIGER, product name: Tiger Microcomputer Food Processor) to about half their original size. The coarsely ground feed pellets (approximately 0.1 kg) and the feed additive (500 mg) were added to a plastic bag and thoroughly mixed in the plastic bag. Then, the remaining coarsely ground feed pellets were added to the plastic bag and mixed further. Feed containing isolate A (feed (A)) was obtained by the above procedure. Feed (A) can be understood as one embodiment of the ostrich rearing composition of the present invention (Example).

[0061] Furthermore, the comparative feed (feed (B)) was prepared using the same procedure as above, except that the dextrin powder (500 mg) was used instead of the above feed additive.

[0062] (3: Ostrich chick rearing experiment) Seventy ostrich eggs laid on the same day were placed in an incubator manufactured by Showa Franki Co., Ltd. (product name: Franki SS-18 for ostriches) and incubated at 36.5°C and 40.0% humidity. Twenty-four chicks (14 males, 10 females) that hatched between July 12 and July 14, 2023, were randomly divided into two groups. The ostrich chicks were raised in a constant temperature and humidity chamber (36.5°C, 40.0% humidity) until rearing in the prefabricated shed described later was started.

[0063] On July 16 of the same year, the ostrich chicks were raised in a prefabricated shed (5.5m long, 3.5m wide, 2.3m high). A 2m x 3.5m section within the shed was divided in half by a partition board (approximately 30cm high), and 12 chicks were raised in each section. A brooder (infrared heater) was installed in each section. In addition, fresh air was constantly drawn into the shed by a ventilation fan.

[0064] One of the two groups (the bacterial cell intake group) was given feed (A), and the other group (the control group) was given feed (B). Both groups had free access to feed and drinking water. The intake period was two months. During the intake period, the weight of each individual was measured 3, 6, 10, 16, 20, 24, 28, and 30 days after the chick was moved to the chick house, and the weight gain from the start of intake was calculated using formula 1 above. The average weight gain for each group was calculated from the calculated weight gain. The results are shown in Figure 2. In addition, the number of surviving chicks was counted at the same time, and the survival rate for each group on that day was calculated using formula 2 above. The results are shown in Figure 3.

[0065] In the group that ingested feed (A), which is the example, the survival rate at 40 days of age was 60% (Figure 3). On the other hand, in the control group that ingested feed (B), which is the comparative example, the survival rate decreased significantly after 15 days of age, and the survival rate at 40 days of age was 0% (Figure 3). Therefore, weight measurements could only be taken up to 35 days of age in the control group (Figure 2). In the group that ingested bacteria, the amount of weight gained at 35 days of age was approximately twice that of the control group. In addition, in the group that ingested bacteria, the slope of weight gain after 40 days of age was steeper compared to before 40 days of age. From these results, it was found that feeding ostriches (e.g., chicks) with bacteria belonging to the species Lactiplantibacillus plantarum (e.g., NITE-BP-04252) can be used to increase weight or improve survival rates. Furthermore, it was suggested that using the feed additives and feed in this embodiment for ostrich farming could increase the production of ostrich-derived food ingredients and significantly contribute to the Sustainable Development Goals (SDGs).

Claims

1. A composition for ostrich rearing containing bacterial cells or bacterial cultures of bacteria belonging to the species Lactiplantibacillus plantarum, or extracts thereof.

2. The ostrich breeding composition according to claim 1, wherein the bacterial cells include heat-sterilized bacterial cells.

3. The ostrich breeding composition according to claim 1 or 2, wherein the ostrich breeding composition contains bacterial cells, and the content of the bacterial cells is 0.0001 to 0.01% by mass relative to the ostrich breeding composition.

4. The ostrich rearing composition according to claim 1 or 2, further comprising basic feed or drinking water.

5. The ostrich rearing composition according to claim 4, further comprising the basic feed, wherein the basic feed comprises protein or lipid.

6. The ostrich breeding composition according to claim 1 or 2, further comprising at least one selected from the group consisting of dextrin, vitamins, trace inorganic salts, organic acids, and enzymes.

7. A method for raising ostriches, comprising the step of administering the ostrich breeding composition described in claim 1 or 2 to an ostrich.

8. The method for raising ostriches according to claim 7, wherein the ostriches include chicks.

9. The method for raising ostriches according to claim 7, wherein the ostrich includes the North African ostrich, Masai ostrich, South African ostrich, Somali ostrich, Mauritanian ostrich, or African black ostrich.

10. A method for producing ostrich-derived food ingredients, comprising the steps of: raising ostriches according to the ostrich rearing method described in claim 7; and processing the reared ostriches into food ingredients.

11. A bacterium possessing a 16S rRNA gene containing a nucleotide sequence that has 99.8% or more sequence identity with the nucleotide sequence described in Sequence ID No. 1, and which has a weight-increasing effect or a survival rate-improving effect on ostriches.

12. The bacterium according to claim 11, having a 16S rRNA gene containing the base sequence described in Sequence ID No.

1.

13. Bacteria deposited under accession number NITE-BP-04252.

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