Composition for oral administration for prevention or treatment of viral infections in fully dosed organism

Abstract

The present disclosure provides an orally administered composition for preventing or treating a disease caused by a virus in an organism of sufficient interest, which contains a cell of Lactobacillus plantarum or a cell culture or an extract thereof.

Description

Technical Field

[0001] The present invention relates to an orally administered composition for preventing or treating viral infections in Decapoda organisms. Background Art

[0002] In recent years, the harm (disease) caused by white spot disease / acute viremia (hereinafter also referred to as "WSD / PAV") has increased in shrimp farms. White spot disease (WSD) is caused by white spot disease virus (WSSV) and is a serious widespread animal disease that affects shrimp farming. WSD / PAV has been reported to occur in China, South Korea, Japan, Southeast Asia (Thailand, Vietnam, Malaysia, Indonesia, etc.), Australia, the United States, Ecuador, etc. When WSD / PAV occurs in a culture pond, a large number of shrimps die rapidly within 3 to 10 days, with a mortality rate close to 100% [Global distribution of white spot syndrome virus genotypes determined using a novel genotyping assay J. Oakey, C. Smith, D. Underwood, M. Afsharnasab, V. Alday-Sanz, A. Dhar, S. Sivakumar, ASSahul Hameed, K. Beattie & A. Crook. Archives of Virology volume 164, pages 2061-2082 (2019)]. In order to suppress the occurrence of WSD / PAV, seed production combining parent shrimp selection by PCR method and fertilized egg disinfection or water quality management of culture ponds is implemented, but it is difficult to prevent the occurrence of WSD / PAV by these measures alone.

[0003] To date, methods for suppressing the damage to the culture of kuruma prawns caused by the infection of white spot disease virus have been developed. For example, Japanese Patent Laid-Open No. 2008-63302 (Patent Document 1) discloses an oral vaccine composition containing a recombinant protein of the outer shell protein of white spot disease virus in shrimp. Japanese Patent Laid-Open No. 6-181656 (Patent Document 2) discloses that yellow head disease is prevented / treated by orally administering a composition obtained by allowing black tiger shrimp to ingest a disrupted product of cells containing bacteria of the genus Brevibacterium. Japanese Patent Laid-Open No. 2015-172019 (Patent Document 3) discloses that the death caused by the infection of white spot disease virus is reduced by feeding lactic acid bacteria fermented products obtained by culturing Enterococcus faecalis NBRC3989 strain with a culture medium containing spent mushroom substrate to prawns. Peraza-Gomez, et. al; Aquaculture Research, (MAR 2011) Vol. 42, No. 4, pp. 559-570. (Non-Patent Document 1) shows that the death caused by the infection of white spot disease virus is reduced by feeding a feed mixed with a probiotic mixture of medicinal plant powder, three strains of lactic acid bacteria derived from tilapia, and one strain of yeast to Litopenaeus vannamei. Partida-Arangure et. al; African Journal of Biotechnology (2013), Volume 12, Number 21, pp. 3366-3375. (Non-Patent Document 2) shows that when feeding shrimp with a shrimp feed further added with white spot disease virus prepared in a feed formulated with a live bacteria mixture (only a 4-strain mixture) of lactic acid bacteria (BAL3 and BAL7) derived from the intestinal tract and hepatopancreas of Litopenaeus vannamei and Bacillus bacteria (BC1 and CIB1) derived from plants (agave); [both are unidentified species], inulin only, and their complexes, the mortality rates of the normal feed application group, the inulin only added group, and the 4-strain mixture only added group are the same, but the mortality rate in the group applying the feed containing the 4-strain mixture and inulin is reduced.

[0004] Prior Art Documents

[0005] Patent Documents

[0006] Patent Document 1: Japanese Patent Laid-Open No. 2008-63302

[0007] Patent Document 2: Japanese Patent Laid-Open No. 6-181656

[0008] Patent Document 3: Japanese Patent Laid-Open No. 2015-172019

[0009] Non-Patent Documents

[0010] Non-patent Document 1: Peraza-Gomez, et. al; Aquaculture Research, (MAR 2011)Vol. 42, No. 4, pp. 559-570.

[0011] Non-patent Document 2: Partida-Arangure et. al; African Journal of Biotechnology(2013), Volume 12, Number 21, pp.3366-3375. Summary of the Invention

[0012] Problems to be Solved by the Invention

[0013] An object of the present invention is to provide an orally-administered composition for preventing or treating viral infections in decapod organisms, which has less impact on the environment and less toxicity to humans, and a method for prevention or treatment.

[0014] Means for Solving the Problems

[0015] The present invention relates to the following [1] to [8].

[0016] [1] An orally-administered composition for preventing or treating diseases caused by viruses in decapod organisms, which contains cells or cell cultures of Lactobacillus plantarum or extracts thereof.

[0017] [2] An orally-administered composition for preventing or treating white spot disease / acute viremia in decapod organisms, which contains cells or cell cultures of Lactobacillus plantarum or extracts thereof.

[0018] [3] The orally-administered composition for prevention or treatment according to [1] or [2], wherein the Lactobacillus plantarum is NITE-BP-03201.

[0019] [4] The orally-administered composition for prevention or treatment according to any one of [1] to [3], wherein the decapod organism is a penaeid organism.

[0020] [5] The orally-administered composition for prevention or treatment according to any one of [1] to [4], which is a feed or a feed additive.

[0021] [6] A method for preventing or treating diseases caused by viruses in decapod organisms, which includes the step of inoculating a composition containing cells or cell cultures of Lactobacillus plantarum or extracts thereof into decapod organisms.

[0022] [7] A method for preventing or treating white spot disease / acute viremia in decapod organisms, which comprises the step of inoculating a composition containing the cells or cell cultures of NITE-BP-03201 or their extracts into decapod organisms.

[0023] [8] The preventive or therapeutic method according to [6] or [7], wherein the aforementioned decapod organisms are organisms of the family Penaeidae.

[0024] Advantages of the Invention

[0025] According to the present invention, there can be provided an oral administration composition for preventing or treating viral infections in decapod organisms and a preventive or therapeutic method, which have less impact on the environment and less toxicity to humans. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 A figure showing (A) the shape of colonies and (B) the results of Gram staining of NITE BP-03201 in Experiment 1.

[0027] Figure 2 A figure showing the inhibitory effect of NITE BP-03201 on decapod infections caused by white spot virus in Experiment 2. DETAILED DESCRIPTION OF THE INVENTION

[0028] Hereinafter, the modes for carrying out the present invention will be described in detail. In addition, the present invention is not limited to the following embodiments. In the present specification, the expression in the form of "A to B" means the upper and lower limits of the range (that is, A or more and B or less), and in the case where no unit is described in A and only a unit is described in B, the units of A and B are the same.

[0029] [Composition for Rearing Decapod Organisms]

[0030] The composition for rearing decapod organisms according to one embodiment of the present invention contains cells or cell cultures of Lactobacillus plantarum or their extracts. The aforementioned composition for rearing may contain cells or cell cultures of Lactobacillus plantarum. The aforementioned Lactobacillus plantarum is preferably NITE-BP-03201. The composition for rearing decapod organisms according to the present invention can prevent or treat infections caused by viruses in decapod organisms.

[0031] ​​NITE-BP-03201 is a bacterium that was deposited under the accession number NITE BP-03201 (original deposit date: April 9, 2020) at the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (NPMD, address: Room 122, 2-5-8 Kamigata-Kamakari, Kisarazu City, Chiba Prefecture 292-0818) based on the Budapest Treaty for international deposit. NITE-BP-03201 is a bacterium that all belong to Lactobacillus plantarum, a kind of lactic acid bacterium. Regarding the mycological properties of the above strain, they are shown in Tables 1 to 2 described later and Figure 1 .

[0032] Since NITE-BP-03201 exists in the natural environment, it is considered to be highly safe when applied in the cultivation of shrimp. NITE-BP-03201 can be an isolated bacterium.

[0033] The composition for culturing decapod organisms according to the present invention contains the cells or cell cultures of the above lactic acid bacteria or their extracts. The cells can be dead cells or live cells. The cells can exist in a culture solution, buffer solution, etc., can be a dried product of the culture solution, or can be a frozen stock solution.

[0034] The cell culture can contain secretions, metabolites, etc. of the bacteria. The cell culture contains peptides, proteins, sugars, enzymes, organic acids produced by the bacteria and the culture media (liquid culture media and solid culture media) containing them.

[0035] NITE-BP-03201 can be cultured according to the usual culturing methods for lactic acid bacteria. As a representative culturing method, a method of culturing at a temperature of 30 °C using MRS (de Man, Rogosa and Sharpe) liquid medium or MRS agar medium is listed.

[0036] The cells or cell cultures are formulated in such a way that the antiviral activity possessed by the cells or cell cultures of the above lactic acid bacteria is not lost. In this specification, the cells or cell cultures of the bacteria are also described as "cell preparations".

[0037] The extracts of the cells or cell cultures are formulated in such a way that the antiviral activity possessed by the cells or cell cultures of the above lactic acid bacteria is not lost. The extracts can be obtained, for example, by subjecting the cells or cell cultures of the bacteria to treatments such as ultrasonic disruption, bead milling, freeze-thawing, chemical dissolution, etc. The extracts can be obtained by salting out, ultrafiltration, ion exchange chromatography of the cells or cell cultures or liquid extraction using an organic solvent, etc. These treatments can be carried out in appropriate combinations. The extracts can contain fragments of the cells of the bacteria, nucleic acids, peptides, proteins, sugars and enzymes. In this specification, the "extracts of the cells or cell cultures" can also be included in the above "cell preparations".

[0038] In this specification, Decapoda, also known as Decapod, is one of the taxonomic groups of crustaceans. Decapoda is a taxonomic group that includes shrimps, crabs, hermit crabs, crayfish, etc. Decapoda includes Dendrobranchiata and Pleocyemata. Dendrobranchiata includes Sergestoidea, Penaeidea, Sicyoniidae, Solenoceridae, Aristeidae, and Benthesicymidae. Penaeidea includes Penaeus monodon (common name: black tiger shrimp), Litopenaeus vannamei (common name: whiteleg shrimp), Fenneropenaeus chinensis, Marsupenaeus japonicas, Melicertus latisulcatus, Metapenaeus ensis, Metapenaeus joyneri, Metapenaeus moyebi, Metapenaeopsis barbata, Metapenaeopsis lata, Penaeus semisulcatus, Trachysalambria curvirostris, etc. Penaeidea contains a large number of important species in the shrimp aquaculture industry. Pleocyemata includes Stenopodidea, Caridea, Polychelida, Achelata, Astacidea, Glypheidea, Axiidea, Gebiidea, Anomura, and Brachyura.

[0039] The composition for culturing decapod organisms can be an orally administered composition. The orally administered composition is not particularly limited as long as it is a composition orally administered to decapod organisms. The composition for culturing decapod organisms can be feed or a feed additive for decapod organisms. The orally administered composition can be applied to the environment (e.g., culture water) for culturing decapod organisms, and the cell preparation of the above-mentioned lactic acid bacteria dissolves in the environment and is orally ingested by the decapod organisms cultured in the environment. The orally administered composition for decapod organisms containing the cell preparation of the above-mentioned lactic acid bacteria can prevent or treat infectious diseases caused by viruses (such as WSSV) in the administered decapod organisms.

[0040] The feed for decapods can contain any components as long as they are components commonly used in the breeding and cultivation of decapod organisms, and can be prepared by any preparation method. The composition of the feed for decapods can be appropriately selected according to the species and growth stage of decapods. For example, the feed for decapods can contain protein sources such as squid powder, krill powder, fish meal, soybean oil residue, and corn bran powder, and binders such as gluten and starch. The feed for decapods can contain known carriers or additives permitted in feeds, and can contain: aquatic organism drugs such as erythromycin preparations, ampicillin preparations, praziquantel preparations, lysozyme chloride preparations, oxytetracycline preparations, spiramycin preparations, nifurstyrenate sodium preparations, lincomycin hydrochloride preparations, flumequine preparations, and glutathione preparations, vitamins such as vitamin C, vitamin B1, vitamin A, vitamin D, and vitamin E, and nutritional supplements such as amino acids such as lysine, methionine, and histidine, pigments such as β-carotene, astaxanthin, and canthaxanthin, minerals such as calcium and silicic acid, trace metals, preservatives, etc.

[0041] The feed for decapods can have any shape and size corresponding to the species and size of the decapods being cultured. For example, the feed for decapods can be a powdered feed obtained by mixing and pulverizing dry raw materials, a solidified feed (dry pellets) obtained by solidifying the powder, a paste-like feed (wet pellets) containing moisture, etc. As an example of the feed for decapods, there is cited the type in which a liquid (such as a culture solution) containing the cells of the above-mentioned lactic acid bacteria is impregnated into a general solidified feed for shrimp culture, and the type in which a powder containing the cells (such as a dried powder of the culture solution) is sprinkled.

[0042] The amount of the cell preparation of the above-mentioned lactic acid bacteria contained in the feed for decapods is not particularly limited as long as it shows antiviral activity against white spot syndrome virus (WSSV), and can be an amount suitable for the culture site, culture temperature, oxygen concentration, culture density, species of decapods as the object, etc. For example, the amount of the lactic acid bacteria cells in the feed for decapods can be 1×10 3 ~1×10 12 cfu (colony forming unit) / g, and can be 1×10 4 ~1×1012 cfu / g, which can be 1×10 5 ~1×10 12 cfu / g. The above-mentioned colony forming units can be determined by the agar plate culture method. The amount of the above-mentioned lactic acid bacteria (content ratio) in the feed for decapod can be 0.001 to 30% by mass, preferably 0.01% to 10% by mass, relative to the weight of the feed. The amount of the bacteria can be weighed with a balance or the like.

[0043] The feed additive for decapod is not particularly limited as long as it can be added to the general feed for decapod farming. The feed additive can be in liquid or powder form and can be freeze-dried. The feed additive can be the cell preparation of the above-mentioned lactic acid bacteria itself. The feed additive can contain a liquid, a spreading agent, etc. for facilitating the attachment, absorption or mixing of the cell preparation of the above-mentioned lactic acid bacteria to the feed for decapod. The feed additive is preferably added to the feed in such a way that the amount of bacteria contained in the feed falls within the above range.

[0044] The feed for decapod containing the cell preparation of the above-mentioned lactic acid bacteria can be fed once a day or in multiple times. The feed containing the cell preparation of the above-mentioned lactic acid bacteria can be fed once every few days. The feeding time of the feed containing the cell preparation of the above-mentioned lactic acid bacteria can be appropriately selected according to the species of the target decapod and the like. The feed containing the cell preparation of the above-mentioned lactic acid bacteria can be continuously fed throughout the entire period of farming (raising), or can be fed only during a part of the period. The "part of the period" can 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 period of raising. In one aspect of the present embodiment, the upper limit value of the above-mentioned "part of the period" is not particularly limited. For example, it can be less than 100% of the entire period of raising, and can be 99% or less. The feeding time of the feed containing the cell preparation of the above-mentioned lactic acid bacteria can be, for example, 1 month to 4 months, or about 3 months. The feed containing the cell preparation of the above-mentioned lactic acid bacteria can be repeatedly fed with continuation and interruption at any time.

[0045] The feed for decapod can be supplied by dropping into the water for raising decapod organisms and can be automatically supplied at regular intervals.

[0046] One embodiment of the present invention is the use of the cells or cell cultures or their extracts of Lactobacillus plantarum in the preparation of a composition for raising decapod organisms. Other embodiments of the present invention are the use of the cells or cell cultures of Lactobacillus plantarum in the preparation of a composition for raising decapod organisms. The above-mentioned Lactobacillus plantarum is preferably NITE-BP-03201.

[0047] [Oral administration composition for preventing or treating white spot disease / acute viremia in decapod organisms]

[0048] One embodiment of the present invention relates to an orally administered composition for preventing or treating white spot disease / acute viremia in decapod organisms, which comprises cells or cell cultures of Lactobacillus plantarum or extracts thereof. The orally administered composition for prevention or treatment may contain cells or cell cultures of Lactobacillus plantarum. The Lactobacillus plantarum is preferably NITE-BP-03201. The orally administered composition for prevention or treatment may be a veterinary drug.

[0049] White spot disease / acute viremia is a decapod infection caused by white spot syndrome virus (WSSV). That is, the orally administered composition for prevention or treatment can be regarded as an orally administered composition for preventing or treating decapod infections caused by white spot syndrome virus. In addition, white spot disease / acute viremia is one of the infections with an increasing incidence of damage (disease) in shrimp farms in recent years. Therefore, the orally administered composition for prevention or treatment can be regarded as an orally administered composition for preventing or treating diseases caused by viruses in decapod organisms. Here, "disease" means damage to aquatic products (such as shrimp) caused by diseases.

[0050] By applying NITE-BP-03201, infections in decapod organisms caused by white spot syndrome virus (WSSV) (hereinafter sometimes referred to as "decapod infections") can be prevented or treated. In this specification, treatment includes alleviation of symptoms, improvement of symptoms, and cure. The lactic acid bacteria have direct antiviral activity against the causative virus of the infection, and the mechanism of inhibiting the infection does not depend on the host. Therefore, regardless of the species or growth stage of the decapod in question, decapod infections can be more generally inhibited. If the above lactic acid bacteria are applied, more stable cultivation of decapod organisms becomes possible.

[0051] As an infection of decapod caused by a virus, WSD / PAV caused by the infection of white spot syndrome virus (WSSV) is listed. When WSD / PAV occurs in juvenile shrimps, the mortality rate is almost 100%, which becomes a fatal problem in the aquaculture industry. As other viruses that cause infections in decapod organisms, it includes Decapod iridescent virus, Taura syndrome virus, yellow head virus, covert mortality nodavirus, Infectious hypodermal and haematopoietic necrosis virus, Macrobrachium rosenbergii nodavirus, myonecrosis virus, etc.

[0052] The prophylactic or therapeutic composition can be an orally administered composition and can be administered to decapod organisms in the form of feed or a feed additive. The prophylactic or therapeutic composition can contain components that can be included in the orally administered composition used in the rearing of the above-mentioned decapod organisms. The prophylactic or therapeutic composition further contains a component known to have antiviral activity against white spot syndrome virus (WSSV) or a component that enhances the immune activity of decapod organisms. The shape, preparation method, and administration method of the prophylactic or therapeutic composition can be the same as those of the above-mentioned orally administered composition. The content of the cell preparation of the above-mentioned lactic acid bacteria in the prophylactic or therapeutic composition is not particularly limited as long as it can inhibit the infection caused by white spot syndrome virus (WSSV) in the administered decapod organisms, and can be in the same range as the content in the above-mentioned orally administered composition.

[0053] One embodiment of the present invention is the use of the cells or cell cultures of Lactobacillus plantarum or their extracts in the preparation of a prophylactic or therapeutic composition for decapod infections caused by white spot syndrome virus (WSSV). Other embodiments of the present invention are the use of the cells or cell cultures of Lactobacillus plantarum in the preparation of a prophylactic or therapeutic composition for decapod infections caused by white spot syndrome virus (WSSV). The above-mentioned Lactobacillus plantarum is preferably NITE-BP-03201.

[0054] [Method for preventing or treating white spot / acute viremia in decapod organisms]

[0055] One embodiment of the present invention relates to a method for preventing or treating white spot disease / acute viremia in decapod organisms, which includes the step of administering the cells or cell cultures of Lactobacillus plantarum or their extracts to decapod organisms. The above-mentioned preventive or therapeutic method may include the step of administering the cells or cell cultures of Lactobacillus plantarum to decapod organisms. The administration can be oral administration or immersion administration. The above-mentioned Lactobacillus plantarum is preferably NITE-BP-03201.

[0056] One embodiment of the present invention relates to a method for preventing or treating white spot disease / acute viremia in decapod organisms, which includes the step of inoculating the cells or cell cultures of Lactobacillus plantarum or their extracts into decapod organisms. The above-mentioned preventive or therapeutic method may include the step of inoculating the cells or cell cultures of Lactobacillus plantarum into decapod organisms. The cell preparation of the above-mentioned lactic acid bacteria can be supplied to decapod organisms as the above-mentioned preventive or therapeutic composition.

[0057] One embodiment of the present invention is the use of the cells or cell cultures of Lactobacillus plantarum or their extracts for the prevention or treatment of white spot disease / acute viremia in decapod organisms. Other embodiments of the present invention are the use of the cells or cell cultures of Lactobacillus plantarum for the prevention or treatment of white spot disease / acute viremia in decapod organisms. The above-mentioned Lactobacillus plantarum is preferably NITE-BP-03201.

[0058] Examples

[0059] Hereinafter, examples are listed to illustrate the present invention in more detail, but the present invention is not limited to these examples.

[0060] [Experiment 1: Isolation and Identification of NITE-BP-03201]

[0061] The isolation source (Ficus hispida) was ground together with sterilized water. The ground liquid was appropriately diluted, added to 1 / 2 MRS liquid medium, and subjected to enrichment culture. The enriched culture solution was spread on MRS agar medium containing calcium carbonate to isolate the microorganisms that formed a halo. 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 does not have catalase activity, it was confirmed as lactic acid bacteria.

[0062] Isolate A was identified by 16S rRNA gene analysis, morphological observation, and physiological / biochemical property tests.

[0063] (1) 16S rRNA Gene Analysis

[0064] Genomic DNA was extracted from isolate A, and the resulting genomic DNA was used as a template. Using the forward primer 9F for cloning and the reverse primer 1510R for cloning (Yoshiyuki Nakagawa et al.: Gene Analysis Method, Method for Determining the Base Sequence of the 16S rRNA Gene, edited by the Japanese Society for Actinomycetes, Classification and Identification of Actinomycetes, 88 - 117 pp., Japanese Society Affairs Center, 2001), PCR amplification of the 16S rRNA gene was performed. The PCR amplification was carried out using Tks Gflex DNA polymerase (manufactured by Takara Bio Inc.), and the amplified product after PCR was purified.

[0065] The cycle sequencing reaction was performed using the purified amplified product after PCR. The cycle sequencing reaction was carried out using the BigDye Terminator v3.1 Cycle Sequencing Kit. The resulting 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, and 1510R were used (Yoshiyuki Nakagawa et al.: Gene Analysis Method, Method for Determining the Base Sequence of the 16S rRNA Gene, edited by the Japanese Society for Actinomycetes, Classification and Identification of Actinomycetes, 88 - 117 pp., Japanese Society Affairs Center, 2001).

[0066] The base sequence of the 16S rRNA gene of isolate A was subjected to BLAST homology search against the microbial identification database DB - BA15.0 (manufactured by TechnoSuruga Lab Inc.) and the international base sequence database (DDBJ / ENA(EMBL) / GenBank) using the microbial identification system "ENKI" (manufactured by TechnoSuruga Lab Inc.). The base sequence of the 16S rRNA gene of isolate A showed 99.93% identity with the base sequence of the 16S rRNA gene of Lactobacillus pentosus (JCM1558), 99.93% identity with the base sequence of the 16S rRNA gene of Lactobacillus plantarum subsp. Plantarum (JCM1149), and 99.73% identity with the base sequence of the 16S rRNA gene of Lactobacillus paraplantarum (DSM10667). However, there was no microorganism with a 16S rRNA gene having a base sequence completely identical to that of isolate A.

[0067] (2) Morphological observation and physiological / biochemical property tests

[0068] The isolated strain A was spread on MRS agar medium and aerobically cultured at 30 °C for 48 hours. The cell morphology, Gram stainability, motility, and colony morphology were observed by the following methods.

[0069] The colony morphology was observed with a stereomicroscope SMZ800N (manufactured by Nikon Corporation). The cell morphology was observed with an optical microscope BX50F4 (manufactured by Olympus Corporation). Favor G “Nissui” (manufactured by Nissui Pharmaceutical Co., Ltd.) was used in Gram staining. 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 the catalase reaction, oxidase reaction, acid / gas production from glucose, and oxidation / fermentation (O / F) of glucose. The API50CHB kit (manufactured by bioMerieux, France) was used to study the physiological / biochemical property reactions of the bacteria.

[0070] As Figure 1 shown in (A) of Figure 1 , the isolated strain A formed round colonies. As Figure 1 shown in (B) of Figure 1 , the Gram stainability of the isolated strain A was positive. The results of the physiological / biochemical property tests and fermentation tests of the isolated strain A are shown in Tables 1 and 2. The isolated strain A was a Gram-positive bacillus that did not show motility and did not form spores. The catalase reaction and oxidase reaction of the isolated strain A were negative, and it fermented glucose. These properties were consistent with the results of the partial base sequence analysis of 16S rDNA and the properties of the genus Lactobacillus, which showed the possibility of belonging. As a result of the fermentation test using the API kit, the isolated strain A fermented galactose, fructose, and melezitose, etc., and did not ferment glycerol, D-xylose, etc. The isolated strain A did not show arginine dihydrolase activity and grew at 15 °C. Among the results of the partial base sequence analysis of 16S rDNA, which suggested belonging to Lactobacillus pentosus and Lactobacillus plantarum, it was different from Lactobacillus pentosus in not showing the fermentation of glycerol and D-xylose, and was consistent with the properties of Lactobacillus plantarum. Therefore, it was known that the isolated strain A was a newly isolated strain belonging to Lactobacillus plantarum. The isolated strain A was internationally deposited as NITE BP-03201.

[0071] [Table 1]

[0072]

[0073] [Table 2]

[0074]

[0075] [Experiment 2: Verification of the preventive or therapeutic effect of oral administration of lactic acid bacteria on WSD / PAV]

[0076] Verify whether the cell culture of lactic acid bacteria administered orally can inhibit the infection caused by white spot disease virus (WSSV) in decapod organisms (i.e., WSD / PAV). As the rearing environment for shrimp, 10 L of artificial seawater was prepared in an 18 L glass aquarium. The shrimp were reared while filtering the rearing water with a SUISAKU 8 of size M. The water temperature was set at 28 °C. 25 Litopenaeus vannamei were used for each test group.

[0077] In the commercially available granular feed used in Thailand, the cell culture of lactic acid bacteria was added to the extent that the feed was sufficiently soaked to prepare an oral administration composition for rearing decapod organisms. The above-mentioned cell culture of lactic acid bacteria was prepared in the following order. First, NITE-BP-03201 was cultured in MRS Broth at a temperature of 30 °C for 1 day to obtain a culture solution. The obtained culture solution was heat-treated at 70 °C for 1 hour, and then dextrin was added so that the final concentration became 4%, followed by freeze-drying. Thereafter, distilled water was added to the freeze-dried cells in an amount equal to the original culture solution, and the cells were suspended to obtain the above-mentioned cell culture of lactic acid bacteria. The turbidity of the original lactic acid bacteria culture solution at a wavelength of 600 nm was approximately 12 OD. The negative control was a liquid prepared by adding dextrin to MRS Broth, which is a medium for lactic acid bacteria, so that the final concentration became 4%, followed by freeze-drying, and then adding distilled water in an amount equal to the original solution and suspending. These were added to the feed and set aside. For feeding, an amount of about 5% of the shrimp body weight was administered 3 times a day. The virus solution used during infection was prepared by grinding Litopenaeus vannamei suffering from WSD / PAV. The oral administration composition was fed starting 14 days before the infection test and continued during the infection test. The virus solution was added to the rearing water, and after soaking for 3 hours, the shrimp were transferred to a new aquarium and observed for 8 days. In addition, the addition amount of the above-mentioned virus solution was determined by a preliminary test in advance, and the amount was such that about 90% of the shrimp died in one week (in this experiment, 5 mL). The survival rate of the shrimp from the start of infection is shown in Figure 2 .

[0078] The survival rate of decapod organisms fed with an oral administration composition (a prophylactic or therapeutic oral administration composition) containing the bacterial cell culture of NITE BP-03201 was increased compared to the negative control. It was shown that the infection (WSD / PAV) caused by white spot syndrome virus (WSSV) can be prevented by orally administering the bacterial cell culture of NITE BP-03201.

Claims

1. An orally administered composition for preventing or treating diseases caused by viruses in decapod organisms, which contains cells or cell cultures of Lactobacillus plantarum or their extracts.

2. An orally administered composition for preventing or treating white spot disease / acute viremia in decapod organisms, which contains cells or cell cultures of Lactobacillus plantarum or their extracts.

3. The orally administrable composition for prophylaxis or treatment according to claim 1 or claim 2, wherein, The Lactobacillus plantarum is NITE-BP-03201.

4. The orally administrable composition for prophylaxis or treatment according to claim 1 or claim 2, wherein, The decapod organisms are organisms of the family Penaeidae.

5. The orally administered composition for prevention or treatment according to claim 1 or claim 2, which is a feed or a feed additive.

6. A method for preventing or treating diseases caused by viruses in decapod organisms, which includes the step of inoculating a composition containing cells or cell cultures of Lactobacillus plantarum or their extracts into decapod organisms.

7. A method for preventing or treating white spot disease / acute viremia in decapod organisms, which includes the step of inoculating a composition containing cells or cell cultures of NITE-BP-03201 or their extracts into decapod organisms.

8. The prophylactic or therapeutic method according to claim 6 or claim 7, wherein, The decapod organisms are organisms of the family Penaeidae.

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