TW201204379A - Compositions for treating or preventing infection of Riemerella anatipestifer - Google Patents

Abstract

The present invention relates to a composition for treating or preventing infection of Riemerella anatipestifer comprising an effective amount of a peptide having a given amino acid sequence and a carrier.

Description

201204379 VI. Description of the Invention: [Technical Field to Be Invented by the Invention] The present invention relates to a composition against infection by R. vaginalis. [Previous technique], the bacterium is the main infectious agent of poultry, and can infect ducks, chickens, cockroaches, swan, cranes, etc. (2〇〇5; 49:1〇4_7). = In the case of ducks, 'Rie's rod g infection is common in young ducks, often accompanied by sepsis septicemia) and sacral serositis (p〇iyser〇si〗 〖), the mortality rate is high, which has a great impact on the duck industry ( Sangha 1985; 29: 128_35; and 1997; 26: 791-802). However, at present, there is limited knowledge of the defense system against duck-resistant pathogens. Even if the vaccine has been developed based on the mutation, it is impossible to produce cross-protection in different blood/month t-like bacteria (cmss-p such as eCti〇n). Therefore, it limits the use of these vaccines in the dyeing of the thief. Therefore, the aquaculture industry is still mainly using antibiotics. (4) Antibiotics used in the bacterium of the genus Rep. include ceftriofbr, lofx (10) fl0xacin, novobiocin (n〇v〇bi〇dn), Lmcomycin, sulphate, ph〇namides, and the like. However, these antibiotics may cause the side 1 to cause drug resistance, and even this resistant strain can be tested by the food chain into the human body, which is detrimental to people's lives.

Vet Diagn invest 2m, 15:26-9 ; ^ Antimicrob Agents Chemother 2005; 49:690-8 ) 〇nemother Park So there is still a need to develop in this area to effectively fight Reed's rods, especially non-antibiotics. In the poultry raising 仏 仏 仏 杆菌 « «. 201204379 SUMMARY OF THE INVENTION The present invention has found that a specific fragment of an antibacterial protein has an anti-staining effect. The invention breaks through the current prevention and control of the Raleigh rod of the poultry culture _ if f has side effects and drug resistance, etc. The antibacterial protein of the "C. sinensis" has a pole 2 for the prevention and control of the Rayleigh rod. Therefore, in one aspect, the invention Provided for use in therapy or comprising: an effective amount of a peptide and a carrier, (1) having a peptide of the amino acid sequence of SEQ ID NO: 1; (=) having a value of, for example, 8 £(^: a peptide of the acid sequence of 2; (3) a peptide having the amino acid sequence of SEQ ID NO: 3; (2 a peptide having the amino acid sequence of SEQ ro NO: 4; and (5) Combination of one or more of the above (1) to (4) - The present invention provides the use of the above-mentioned winning form for the preparation of a music for treating or preventing the infection of the genus of the genus of the genus. The killing of the Rayleigh rod is used for the preparation of the composition containing the above-mentioned peptide and the carrier in an effective amount, and is applied to the article or environment containing the R. cerevisiae of the month b. Having a detailed description of the various specific embodiments and the patented target of the present invention will be clearly presented below. The general knowledge in the field of practice does not require further - 兄,, in the case of the following, the invention can be used according to the description herein to its broadest scope. The following description should be taken as an illustration and not as Any means is a limitation of the limitations of the present invention. [Embodiment] The technical and scientific terms of Luming iff qing qing 'No SHI lang lang are the same as those commonly understood by those skilled in the art of this technology. The article "-" refers to a grammatical term of the article H or more (ie, at least one of 201204379). In one aspect, the present invention provides a composition for treating or preventing a Rayleigh rod, which includes an effective amount a peptide and a carrier selected from the group consisting of a sputum composition (1) a peptide having the amino acid sequence of SEQ ID NO: 1; (2) an amino acid sequence having SEQ ID NO: 2. a peptide; (3) a peptide having the amino acid sequence of SEQ ID NO: 3; (4) a peptide having the amino acid sequence of SEQ ID NO: 4; and (5) one of the above (1) to (4) Or a combination of multiples. - In a further aspect, the invention provides a method for killing a bacterium of the bacterium The method 'includes an effective amount of the above-mentioned peptide and face composition for use in an article or environment that may contain R. cerevisiae. The "protein", "polypeptide" or acesulfon peptide used in the text may be interactive. By use, it refers to a biological polymer obtained by linking amino acids via peptide bonds. A protein or polypeptide of a specific sequence can be prepared by chemical synthesis or genetic engineering techniques. The peptides according to the present invention each have SEQ ID N0: The aminoguanidine sequence of 2, 3, or 4 can be prepared by a conventional chemical synthesis method or genetic engineering technique and can be prepared according to the teachings and teachings of the present invention. Phase-specific technology, including polymerase chain reaction

(PCR) amplification techniques, nucleic acid purification, plastids, enzyme processing of nucleic acid fragments, sequencing techniques, and protein expression and synthesis, see M〇lecular cl〇ning : A

Laboratory Manual ' Second Edition ' Cold Spring Harbor Laboratory

PreSS ’ Sambr〇〇k J et al., 1989, and Current Protocols in

Molecular Biology, edited by Frederick M. A. et al., 2001, John Wiley & Sons, Inc. α The peptide according to the present invention can also be isolated from a living organism (for example, fish or shrimp) and the example of the deficiency can be separated from the grouper (Epinejaus (3) (6) plus) to isolate the grouper antibacterial protein-1 (epinecidin). -1) which has the amino acid sequence of SEQ ID NO: 1 (see, for example, Nai-Ce//〇 〇/2007; 26: 403-13). In addition, 201204379 can isolate an anti-lipop〇ly Saccharide fact〇r (ALF) from grass shrimp (household (9) from (10)), which has an amino acid sequence of SEq ID N〇2; The endotoxin of the fresh-negative g, including a chemostructure or a cyclic structure, wherein the cyclic structure is produced by the C-terminal cystine acid of the anti-lipopolysaccharide factor and the n-terminal cystic acid. Formed by the key (see / ζ ζ · dirty (10) ρ / ί 2αα? / 2〇〇7; 7: 687-700). In one embodiment, the linear structure of the molecule is placed in a 1% by weight dimethyl sulfazone (DMS® Dimethyl Sulf0xide), and the cyclic structure of the anti-lipid oligo is obtained after a reaction at room temperature for several hours. In addition, hepatidin can be isolated from Wu Guoyu (7) Tengxin (10)^biliary-), including TH1-5 and TH2-3' having the amino acid sequences of SEQ ID NOS: 3 and 4, respectively. (See Mol immunol 2007; 44: 1922-34) 胜 The peptide according to the invention includes variants, which refer to peptides that are essentially homologous to the original peptide, but due to one or more deletions, insertions or The substitution results in a seric acid sequence which is different from the peptide of the present invention, but retains the same biological activity as the original. The substitution may include a sequence of retention substitutions, meaning that the amino acid group is known to be substituted with a group having similar physiochemical properties. In ancestors or eggs, the appropriate heterogeneous substitution of 'silveric acid' is well known to those of ordinary skill in the art and can be accomplished without altering the biological enthalpy of the resulting molecule. Preferably, the variant has the same steric structure as the original protein peptide. ^文中赖 "Active" B_ means having the effect of confrontation. The activity can be demonstrated by the Minimization Inhibition Concentrations (MIC) for R. cerevisiae according to the US Clinical Microbiological Laboratory Standards. The MIC of the present invention has a MIC of about 6.25_5Q, as shown in the following Table 1, and the cyclic structure of the amino acid sequence of & NO. The hall of the peptide is about mm, the MIC of the linear structure peptide is about 6.25. • The MIC of the amine phase of SETI^a 3 is _ 25 = month ^ and the silk phase of the SEQ m NO: 4 of the present invention Win = 201204379 is 100-450 pg/ml. Or pre-existence (4) therapeutic treatment τ to prevent the disease from the hair == two-body infected with the bacterium of the bacterium, or the sensation of the peptide, until the individual may be exposed to the above-mentioned individuals may contact the bacillus and the tendency The individual or individual of the above-mentioned therapeutic peptides is subjected to the body weight and variety of the treated body, the means of receiving the agent, and the preparation according to the disclosed agent: the acceptable indication The active ingredient contained in the composition is compatible, and is intended to be administered to a body or an avid ingredient, and == when the carrier is adjusted to a carrier, the drug group & And acceptable to the excipients, for example, examples of the emulsifiable carrier include but not powder, gum arabic, squaric acid =, f recommended sugar, sorbose, mannose, Calcium, microcrystalline cellulose, Polyethylene buckwheat melon, Astragalus membranaceus, Minglu, Lixi acid methylcellulose; Lubrication;: Second, Guanjing; Emulsifying and suspending agent; with Dendrobium, stearic acid Magnesium and mineral oils; wet sweeteners, and flavoring agents, for example, benzoic acid (iv) and vinegar, pills, pills, powders, acids, packs, suppositories, injections and packaging Stinger, syrup, soft and hard gelatin capsules,. Knife. In a particular embodiment, the composition of the invention 7 201204379 is in the form of an injectable solution. The intestinal tract is administered by any physiologically acceptable route, for example, orally, meat, intravenously, subcutaneously, intraperitoneally, transdermally, rectally, or by injection. In a particular implementation, the composition of the invention is

In the environment of :=, (for example, 'avian: long = field = domain), used to control infection by R. brevis. N 1杲2 day ^^ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ , with excellent / alpha therapy or to prevent the effect of Reptile infection. The minimum bacteriostatic action of the concentration of smutr against the bacterium of the genus Rep. in a special case, the test for the common cultured ducks confirmed that the cockroach and the fine g of the present invention were applied in advance, and the hair was applied in advance. _ wins the dye again, or first accepts the bacterial infection and then the peptide of the present invention, the survival rate of the young duck is significantly higher than that of the control group only. Further, the invention provides a method of killing a bacterium of the bacterium, which comprises administering an effective amount of the above-mentioned peptide and a fine composition to an article or environment which may contain a bacterium. In a specific example, the treatment of the bacterium with the peptide of the present invention, using a transmission electron microscope, clearly observes that after the treatment of the bacteria, the cell membrane exhibits an irregular shape and is damaged, resulting in cell biliary cells such as cells. Causes cell death. In a specific example, the composition of the present invention is formulated into a liquid form, a granule form, or a powder form. In particular, the compositions of the invention may be applied to the farm or farmed waters of the animal. The present invention will be further illustrated by the following examples, but the actual invention is not limited to the embodiment shown. 201204379 Example 1: Synthesis of the anti-g protein of the present invention (4) The present invention was prepared by the Gil Biochemical Company (Shanghai, China) using a solid phase procedure of Fmoc/tBu chemistry. Table 1 lists the names of the individual anti-prion proteins and the amino acid sequence. Table 1

Name Grouper Antibacterial Protein-1 Amino Acid Sequence GFIFHIIKGLFHAGKMIHGLV (SEQ ID NO: 1)

Grass shrimp circular anti-lipopolysaccharide molecule ECKFTVKPYLKRFQVYYKGRM (CA) WCP (SEQ ID NO: 2) - a circular structure of grass shrimp linear anti-lipopolysaccharide molecule ECKFTVKPYLKRFQVYYKGRM WCP (SEQ ID NO: 2) - linear structure Wu Guoyu liver antibacterial Peptide (TH1-5) Wu Guoyu Liver Antibacterial Peptide (TH2-3) GIKCRFCCGCCTPGICGVCCRF (SEQ ID NO: 3) QSHLSLCRWCCNCCRSNKGC (SEQ ID NO: 4) The synthesized peptide is extracted, lyophilized, and utilizes a reverse phase high-performance solution. Purification by reverse-phase high-performance liquid chromatography (RP-HPLC). The molecular weight and purity of the purified polypeptide were analyzed by mass spectrometry and high performance liquid chromatography. The purified polypeptide was reconstituted with phosphate buffer (pH 7.4) for subsequent experiments. Example 2: Determination of Minimum Inhibitory Concentration This example tests the minimum inhibitory concentration of antibacterial proteins against several strains of R. eutropha, including A2, B2, T6, RA16 provided by Professor Yu Zhangyou of the Department of Veterinary Medicine, National Chiayi University. And the T10 strain, the CFC27, CFC437, CFC363, RA3 and RA9 strains provided by Prof. Zhang Zhaofu from the National Taiwan University Veterinary Research Institute, and the MRS isolated from the cherry valley duck of the private duck farm in Jiaoxi, Taiwan. Strain. The culture solution containing each strain was combined with 201204379 687 7〇m.矣i & 枰友进仃 (/咐7(10)2007; 7: 687-700). Table 2 shows the test results. Table 2

CFC437 CFC363 RA3 RA9 CFC27 RA16 T10 B2 T6 A2 MRS House bacteriostatic repellent (lug/ml) ^--LA_EPI TH1-5 Th2-3 25 12.5 12.5 12.5 25 12.5 12.5 12.5 12.5 25 25 12.5 6.25 12.5 12.5 25 12.5 12.5 12.5 12.5 12.5 25 25 50 25 50 25 25 25 6.25 25 25 25 50 50 25 50 100 25 50 25 400 50 50 ND 100 ND 100 100 100

450 ND ND As indicated by f 2, the above antibacterial protein has antibacterial effect against various strains of Listeria species listed. The minimum inhibitory concentration range measured is 6 25_45 〇 μέ/ηι1. Example 3: Transmitted Electron Microscopy The experimental procedure for a transmission electron microscope is the same as that of the previously published literature 2007; 7: 687-700). Briefly, the bacterial cultures of the strains of Reuteri strains T6, MRS and CFC27 were passed through a centrifuge (Beckman Coulter, Avanti J-20XP, JA-25.50, Palo Alto, CA, USA) at 3000 rpm at 4 °. Centrifuge for 5 minutes at C. The precipitate was reconstituted in PBS at a concentration of 1.0 at 550D550 nm. 5 ml of the bacterial reconstituted product was mixed with various antibacterial proteins (1 μ μβ/πι1) and reacted for 6 hours. The control group contained only bacterial reconstituted material (not treated with antibacterial protein) suspended in PBS or culture. After sectioning, observation was carried out by a transmission electron microscope (Hitachi, 201204379 H-7000, Tokyo, Sakamoto) under conditions of 75 kV. CA, =rTHf ^ It is clearly observed that the two strains of the R. eutropha strain treated with the antibacterial protein® and rFrr? H1-5 of the present invention have a cell type of 2 (Fig. 1C) and an untreated control. Groups, in the fine and broken, cause the cells to flow out of the cells, causing cell death. Example 4: Animal Infection Experiment Animals were tested with pumpkin duck (C(iv)-dUCk) and Cherry Valley Duck. The source of the animal is the feed and sterile water that can be obtained by the four-day hatchery in the general farm. The young ducks are kept in the isolation and the temperature is maintained in the isolation room. Use ° (G x 1/2' Terumo Medical, Elkton, MD, USA) to inject Ray's white and / or PBS_ into the abdominal cavity of the duck, each time the band ^ ^ real 8 test" each group 3G only tested juvenile duck 'control group received: each: solid (Xl0 colony forming unit (cfu) 'every duck) * PBS injection; oysters η group shell! Receive each antibacterial protein (100 飓, each duckling) Injection with a mixture of Lei's rod + x 10 cfu 'each duckling'. The shooting experiment was 6 immediately after the first experiment was completed for 336 hours, and additional injections of Frey's f bacteria (ΐχΐ.6*, each duckling) were added. In the three f experiments, there were 20 tested ducklings in each group, first injection of Rees: xio cfu, mother only ducklings, and then 2 or 24 hours later, pBS or each antibacterial egg was injected 100 tons, each Young ducks. f In the fourth group of experiments, each group had 2 babies tested, first injected pBs antibacterial protein (10), each duckling), and after 2 or 24 hours, skin shot, rod 8 ((10), each duckling.) Animal test, in the Cherry Valley duck, the strain of the bacterium used in the bacterium is 'EPI, CA, LA, and TH1-5, and anti-201204379 3 ί The interval between the bacteria is 2 hours; in the pumpkin duck, the used Rex ## strain is Τ6 'The antibacterial proteins tested are EPI, CA, LA, and THW' and the antibacterial protein and the bacterium of the bacterium The interval was 24 out and the statistical analysis was performed on each of the sisters' experimental data to compare the differences between the two groups. The comparison between the groups was tested by the variation f (ANOVA) of the spss software. The different letters on the graph are shown in There is a difference between the two groups, and (4) the letter ugly does not exist between the two groups. The results show that EPI, CA, LA, and THM are based on the truth. And iHl-5 can significantly reduce the mortality of the mrs sensation: the mortality of the peach duck (Figure 2); while Ερ, CA, LA, thi_5, and ΤΗ2-3 can significantly reduce the death of the pumpkin duck infected by Ding 6 Rate (Fig. y. = 2 (A), the cherry valley duck in the road 8 strain (丄χ 1〇8 cfu / duck) Sense = 14 days after the death rate is extremely high (in 3 days, 3 animals in the animal) 23 deaths) 'but if the antibacterial protein LA or THl-SdOOpg/ml is applied together and the 14-day survival paste of the animals of the 8 strains reaches 93.33% (2 out of 30 animals in three days), the joint application The 14-day survival rates of the animals with antibacterial protein Ερ or CA and strains also reached 7666% (7 out of 3 animals in 9 days) and 73.33% (8 out of 30 animals in 9 days). In the experiment of jointly applying antibacterial protein and bacteria, the survival rate of the young ducks was only in the control group of the bacteria (?<〇.〇5). As shown in Fig. 2(B), the animals were subjected to the above-mentioned joint application. After playing antibacterial protein and bacteria, after 14 days, the mrs strain (lxl〇8cfu/duck) was applied again. The fruit showed that the survival rate of only the ducklings that received the bacteria injection was only 166.6%. The survival rate of the young ducks with the antibacterial protein EPI, CA, LA, or TH1-5 and bacteria was increased to 47.82%, 63 63%, 71 42% and 75%, respectively. As shown in Figures 2(C) and 2(D), the survival rate of young ducks was significantly higher than that of the beatings, whether they were first infected with bacterial infection and then treated with anti-8 protein. Bacterial and PBS control group. * Similarly, as shown in Figure 3(A), the survival rate of pumpkin squash in T6 strain (1 X l〇8cfll/duck) was only 30%, and only the pBS ducklings 201204379 TH2 ί动 Λ Λ 抗菌 抗菌 EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP EP %. In the Wei and fine _ experiments, the survival rate of the young ducks was significantly higher than that shown in Figure i (B). After the pumpkin ducks received the above-mentioned joint application against f white and =, 'after 14 days', the τ6 strain was again applied. 〇8 - :1 public: The fruit shows that the young ducks that only receive fine limbs have a survival rate of only 4%, while the young ducks that have received the common antibacterial protein TH2_3, Epi, Tm_5, la, or bacteria The survival rate is higher, 91.7%, 90%, 86.70/0, 86.7%, and 83.3%, respectively. In addition, as shown in Figure 3 (C), only animals that have been beaten by the Τ6 strain will be caused within days. High mortality rate (2 deaths out of 2 animals in 4 days); OBJECTIVE: If the antibacterial protein Ερι TH2-3, TH1_5, LA or CA is applied 24 hours after the snoring 6 strain, the animal survives. The rates are higher, respectively, %, 95%, 90%, 85%, and 80%. Finally, as shown in Figure 3 (D), the animals that received only T6 strains had a mortality rate of 14 days. 80%; in contrast, if antibacterial protein fertilizer, C^A, LA 8, ΤΗ1-5 and ΤΗ2-3 (1〇〇μβ) were applied in advance, the strain 6 (1x10 cfU/duck) was applied after 24 hours. ), then animals The survival rates were higher, 7〇%, 〇5%, 60%, 55%, and 65%, respectively. Example 4: Bactericidal activity test of antibacterial protein in animal liver—The ducklings received different forms of antibacterial protein and/or The strain was applied. In the first group of tests, the animals were first beaten with T6 strain (ixio8 cfil/duck), and after 24 hours, the antibacterial proteins EPI, CA, LA, TH1-5 or TH2-3 were applied (1〇 〇μβ/ duck). In the second group of experiments, the animal first applied antibacterial protein Epl, CA, LA, TH1-5 or ΤΗ2-3 (100μδ/duck)' 24 hours later and then applied T6 strain (lxl08 cfu/ Duck). In the third group of experiments, 'animals were combined with T6 strain (ixl08cfll/duck) and anti-® protein EPI, CA, LA, TH1-5 or TH2-3 (100 pg/duck). 201204379 Accepted in animals After 1, 3, 5, and 7 days, liver tissue was taken for analysis. First, the liver tissue was homogenized and placed in PBS, which was spread on a sheep blood agar plate, and then the plate was placed on the plate. 37. (: 24 hours of culture, and finally calculate the number of bacterial colonies separately. Calculate the significant difference using the SAS analysis program (ρ < 0·05). The results show that no The antibacterial proteins EPI, CA, LA, TH1_5 4ΤΗ2_3 ^ can be significantly reduced below the bacteria after 7 days of injection with the bacteria (Fig. 4Α), post-infection (Fig. 4Β), or pre-infection (Fig. 4C). Number of bacteria in the liver (ρ < 0.05). Summarizing the above experiments, the present invention confirmed the antibacterial proteins EPI, ca, LA,

TH1-5 or TH2-3 has bactericidal activity against R. vaginalis and can significantly increase the survival rate of infected animals. In view of the increasing side effects of antibiotics and the increasing number of drug-resistant strains, the present invention confirms that the above-mentioned antibacterial proteins have the potential to develop into infections against R. vaginalis, and agents or adjuvants, especially in the waterfowl breeding industry, have a . Those who are familiar with the art can not confuse the duck god. It should be noted that the present invention is not limited to = there is a change in other drawings according to the application. :=,本发====式列微镜影像:=蛋=, penetrating, untreated MAS strain, antibacterial protein to (the illusion is not treated by antibacterial protein CA treatment ^^^ 1 treated strains, strains, and treated mice with antibacterial egg = (1) are _ strains at M = f, and (1) to () (repeated test results under the same conditions of sputum. Figure 1 14 201204379 for CFC27 strain (4) to (4) are untreated CFC27, strains, CFC27 treated with antibacterial protein Cong, CFC27 treated with m27; anti-protein LA, CFC27 strain treated with chitosan TH1-5, and (7) to repeat the test results under the same conditions of 〇. Fig. 1 (C), (4) to (〇 are untreated T6 strains, respectively, J = Ϊ Τ6 strain, Τ6 strain treated with antibacterial protein CA, = protein LA treated Τ6 strain, and treated with antibacterial protein τηι_5 for tongue, - strain, and (1) to (1) are at (a) to (e) Repeated test results under the same conditions. Figure 2 below shows that the antibacterial protein of the present invention can increase the survival rate of cherry valley ducks which are affected by the bacterium of the bacterium R. serrata. In Fig. 2(A), Purchased strains; deleted PBS; EPI,

The table does not jointly apply various antibacterial proteins and R. smegmatis. In the same condition of Figure 2 (B Ξίί表, 2 (A), additional injections of R. vaginalis, σ fruit. In Figure 2 (C), RA-2hr-PBS is first administered with R. Then, after 2 hours, the PBS, RA-2hr-THl-5 was firstly applied to the bacterium, and then the antibacterial protein TH1_5 was applied after 2 hours; from the Qin LA, the arsenic was first applied, in 2 hours. After the application of antibacterial protein LA; ^_2111^ eight is first applied to thunder, bacillus, and then applied antibacterial protein CA after 2 hours; and is first applied to the bacterium, and then 2 days after the application of antibacterial protein Ε π. 2 (9), PBS-2hr-RA was first administered with PBS, and then 2 days after the application of Rep.; THl-5-2hr-RA was first applied antibacterial protein Tm_5, and then 2 days after the application of Rep; LA-2hr-RA is first applied antibacterial protein LA', and then treated with R. eutropha 2 hours later; CA_2hr-RA is first applied antibacterial protein CA, 'After 2 hours, Streptococcus is administered; and muscle is first The antibacterial protein EPI' was applied to the bacterium, and the antibacterial protein of the present invention was able to increase the survival rate of the pumpkin duck infected by the bacterium R. serrata 6 in Fig. 3 (Fig. 3 In A), ra means that only the Rayleigh rod halogen is applied, PBS means that only PBS is applied; EPI, CA, LA, TH1-5, and TH2-3 15 201204379 indicate that the respective antibacterial proteins and R. eutropha are jointly applied. In 3 (B), each line indicates the result of additional injection of R. eutropha under the same conditions as in Fig. 3 (A). In Fig. 3 (C), RA-24hr-PBS is first administered with R. rapae, and then After 24 hours, PBS was applied; RA-24hr-TH2-3 was firstly administered with R. rapae, and then the antibacterial protein TH2_3 was applied after 24 hours; it was first applied to T. reesei and then after 24 hours. Protein Tm_5; is first applied to the bacterium, and then the antibacterial protein LA is applied after 24 hours; RA-24hr-CA is first applied to the bacterium, and then the antibacterial protein CA is applied after 24 hours; and RA-24hr-EPI The antibacterial protein EPI was first applied after 24 hours. In Figure 3 (D), 'PBS-24hr-RA was first administered with PBS, and after 24 hours, the strain was killed; TH2_3_24hr- RA is the first to fight the anti-peptone 2?, and then after 24 hours, the bacterium is killed; after the 丨 伽 伽 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋LA-24hr-RA is first applied antibacterial protein la, and then treated with R. eutropha 24 hours later; CA-24hr-RA is first applied antibacterial protein CA, then 24 hours after the application of R. serrata; and EPI -2hr-RA is first applied to the antibacterial protein EPI, and then administered to the bacterium, 24 hours later. Figure 4 shows the results of the bacteriostatic activity of the antibacterial protein against the R. reuteri T6 strain in the liver of the animal. In Fig. 4(A), RA indicates that only R. serrata was administered;

And EPI+RA, CA +RA, LA +RA, ΤΗ1 -5 +RA, and ΤΗ2-3 +RA indicate that each of the antibacterial proteins and R. smegmatis are co-administered. In Fig. 4 (β), it is indicated that only R. serrata is administered; RA-24hr-TH2-3 is first applied to R. rapae, and then the antibacterial protein TH2_3 is applied after 24 hours; RA-24hr-THl-5 is the first application The antibacterial protein TH1_5 was applied after 24 hours; the antibacterial protein la was first applied after 24 hours; the RA-24hr-CA was first applied to the bacterium, and after 24 hours The antibacterial protein CA was applied; and the RA-24hr-EPI was firstly administered with R. eutropha, and the antibacterial protein EPI was applied after 24 hours. In Fig. 4(C), RA indicates that only R. serrata was administered; TH2-3-24hr-RA was first applied to the antibacterial protein TH2_3, and then T. reesei was administered after 24 hours; THl-5-24hr-RA was first The antibacterial protein TH1-5 was applied, and then the bacterium was killed by 24 hours after 16 201204379; LA-24hr-RA was first applied with antibacterial protein la, and then treated with R. eutropha 24 hours later; CA-24hr-RA was the first The antibacterial protein CA was applied, and the R. eutropha was administered after 24 hours; and the EPI-2hr-RA was first applied with the antibacterial protein EPI, and then the R. eutropha was administered 24 hours later.

17 201204379 Sequence Listing <ιι〇> Central Research Institute

<120> Composition for treating or preventing Reye's infection; <130> XA0001/ACA0034TW <160>

<170> Patentln 3.5itS

<210> 1 <211> 21 <212> PRT <213> spotted grouper (Epinephelus coioides) <400>

Gly Phe lie Phe His lie lie Lys Gly Leu Phe His Ala Gly Lys Met 1 5 10 15

Lie His Gly Leu Val <210> 2 <211> 24

<212> PRT <213> Grasses (Panaeus monodon) <400> 2

Glu Cys Lys Phe Thr Val Lys Pro Tyr Leu Lys Arg Phe Gin Val Tyr 1 5 10 15

Tyr Lys Gly Arg Met Trp Cys Pro 20 <210> 3

<211> 22 <212> PRT <213> Wu Guoyu (Oreochromis mossambicus)

<400> 3

Gly lie Lys Cys Arg Phe Cys Cys Gly Cys Cys Thr Pro Gly lie Cys 1 5 10 15

Gly Val Cys Cys Arg Phe 20 <210> 4

<211> 20 <212> PRT <213> Wu Guoyu (Oreochromis mossambicus) <400〉 4

Gin Ser His Leu Ser Leu Cys Arg Trp Cys Cys Asn Cys Cys Arg Ser 15 10 15

Asn Lys Gly Cys 20 1

Claims (1)

201204379 VII. Patent application scope: : 9ΑΛ 4 into 'oral therapy or prevention of trespassia (and β(10), therefore less than r) Do not include an effective amount of peptide and carrier, the peptide is selected from The following (2 peptide having the acid sequence of SEQ ID NO: 1; peptide 2 having the seric acid sequence of SEQ ID NO: 2) having the amino acid sequence of SEQ ID NO: 3 = having SEQmN: 4 a peptide of the amino acid sequence; and a combination of one or more of the above (1) to (4). Xrrv The composition of claim 1, wherein the peptide having the amino acid sequence of SEQ ID. has a linear or cyclic structure. A composition of a poultry 3, Qianlin for treating or preventing the composition of the third aspect of the patent, wherein the poultry is a duck. The alfalfa is selected from the group consisting of the following peptides for preparation for treatment or prevention of Rees Use of the composition of the genus-infected product: (1 ^ a peptide having the amino acid sequence of SEQ ID NO: 1; a peptide having the amino acid sequence of SEQ ID NO: 2; a peptide of the amino acid sequence of SEQ ID NO: 3; ^4) a peptide having the amino acid sequence of SEQ ID NO: 4; and (5) a combination of one or more of the above (1) to (4). The use of the fifth item of the claim, wherein the peptide having the amino acid sequence of seq 1 is linear or cyclic. The application of the fifth aspect of the patent application is in which the composition is used. For the treatment or prevention of F. faecalis infections in poultry. 8. According to the use of item 7 of the scope of the patent application, wherein the poultry is a duck. The method of killing Lei's off, which is turned over to the secret material, Included in a composition comprising a peptide selected from the group consisting of a carrier selected from the group consisting of: Environment: & ", with 201204379 (1) a peptide having the amino acid sequence of SEQ ID NO: 1; (2) a peptide having the amino acid sequence of SEQ ID NO: 2; (3) A peptide having the amino acid sequence of SEQ ID NO. 3 (4) having an amino acid sequence as SEQ ID Na4, to i4) wherein - or a combination thereof. The method of claim 9 of the patent application scope, wherein the amino acid sequence of the NO: 2 amino acid sequence has the same formula as SEQ K 11. It is liquid or granular according to the ninth item of the patent application scope. Or powder. The method, method, and composition described herein are 酉 12. 12. According to the scope of patent application No. $ or cultured waters. , method, and environment described in the culture 2