KR101099629B1 - A novel Newcastle disease virus K148/08 and a vaccine for Newcastle disease comprising the same - Google Patents

Abstract

The present invention relates to a vaccine for poultry protection against diseases caused by Newcastle disease virus infection, comprising the Newcastle disease virus deposited under accession number KCTC 11570BP and the Newcastle disease virus of the present invention and a pharmaceutically acceptable carrier or diluent.

Newcastle disease, Newcastle disease virus, vaccine

Description

A novel Newcastle disease virus K148 / 08 and a vaccine for Newcastle disease comprising the same}

The present invention relates to a novel Newcastle Disease Virus K148 / 08 strain, and a Newcastle Disease vaccine containing the virus.

Over the last few decades, the production of commercial edible chicken has evolved into an industry characterized by high efficiency in livestock raising. However, a strong tendency to increase the efficiency of the breeding situation cannot be achieved without overcoming some essential difficulties. The most significant increase in the incidence of infectious diseases that occurs frequently in animal populations confined in high density, closed spaces. Many of the most destructive diseases in poultry have been controlled or limited by treatment or vaccination with therapeutics such as antibiotics. Unfortunately, there are still numerous diseases of complex etiology that are not controlled by drugs and do not have adequate vaccination programs.

Newcastle disease (hereinafter referred to as "ND") is an acute infectious disease of chickens caused by the ND virus (Newcastle disease virus, hereinafter referred to as "NDV") and is classified as a Type 1 statutory infectious disease in the outbreak of livestock epidemic. It is a disease that has a duty of killing and reporting to the authorities in case of a disease, and is the only reportable disease designated by the Office International des Epizooties (OIE), which occurs only in Korea. Infection of chickens with NDV causes clinical manifestations such as depression, green diarrhea, speculum and conjunctivitis, with up to 100% mortality in non-immunized chickens and up to 50 in immunized chickens. It has been reported to cause mortality up to%.

NDV is susceptible to most birds and particularly high poultry including chickens, pigeons, crows and parrots, and is known to cause conjunctivitis in humans. On the other hand, wild birds such as ducks, geese, and jaws are able to infect NDV but have been reported to be more resistant than other birds.

NDV was first isolated by Doyle in 1926 in poultry in Newcastle, England, and was named NDV. Paramyxoviridae Avian paramyxovirus type-I (APMV-I) virus belongs to Avulavirus . NDV is a non-segmented single-stranded RNA gene consisting of L (large), HN (haemagglutinin-neuraminidase), F (fusion), NP (nucleoprotein), P (phosphoprotein) and M (matrix) proteins. HN and F proteins are known to play an important role in the pathogenicity of ND.

NDV is classified into velogenic strain, mesogenic strain, lentogenic strain and apathogenic strain according to the virulence of the virus based on the mortality rate for chicks. It is divided into eight genotypes. Genotypes II, III, and IV are known to be genotypes isolated during the first NDV pandemic, and genotypes V and VI NDV are known to be associated with the second pandemic in the late 1960s. Since then, genotype Ⅶ and NDV have spread to some parts of Europe, including Asia and South Africa, and to date have caused significant damage to the poultry industry worldwide. In order to prevent NDV infection, mid toxin, attenuated and non-pathogenic strains are used as vaccines among NDV outdoor isolates isolated from various regions, and attentive and non-pathogenic strains are used as vaccines in Korea.

Since the first occurrence of NDV in Korea was confirmed in 1926, various genotypes have been reported continuously for 80 years. Since 1995, the prevalence of genotype Ⅶ NDV has been confirmed. Therefore, as a part of 'prevention measures for minimizing the damage of Newcastle disease in Korea' since the 1990s, NDV vaccine against one-day-old herbivorous lettuce is legally mandated in domestic hatcheries. In addition, only a weak and non-pathogenic strains are used to minimize the vaccination response. Among them, intestinal affinity strains are used rather than respiratory affinity strains to minimize the effects on the respiratory tract.

The domestic ND live vaccine market forms a relatively large poultry vaccine market size of 4 billion won as a single vaccine, and the types of live ND vaccines used in Korea vary as shown in FIG. 1. However, in the ND live vaccine market, two multinational company-owned products currently have 80% market share, and no domestically developed and produced vaccines exist.

In the present invention, to test the safety and efficacy when spraying the K148 / 08 strain NDV isolated from domestic wild ducks compared to the existing commercial ND vaccine strain (see Figure 1) to develop a live poison vaccine applicable to domestic 1-day-old herbivorous It was.

The present invention solves the above problems, and the object of the present invention is to isolate a novel Newcastle disease virus to be used as a live New vaccine vaccine.

Another object of the present invention is to provide a live vaccine of Newcastle disease.

Yet another object of the present invention is to provide a method for producing Newcastle disease virus.

Yet another object of the present invention is to provide a method for preparing a Newcastle disease vaccine.

Another object of the present invention is to provide a method for controlling a disease state resulting from Newcastle disease virus infection in poultry.

In order to achieve the above object, the present invention provides Newcastle disease virus deposited with accession number KCTC 11570BP.

The virus of the present invention was deposited on October 8, 2009 with accession number KCTC 11570BP to the Korea Institute of Bioscience and Biotechnology, Yuseong-gu, Daejeon, Korea.

In one embodiment of the present invention, the virus is preferably isolated from domestic wild ducks, but is not limited thereto.

A detailed description of the isolation method of Newcastle disease virus of the present invention is disclosed in Example 1.

As a preferred starting material in the isolation method of the virus of the present invention, it can be isolated from diseased algae, in particular from the respiratory tract of wild ducks or excreta excreted by such birds. It should not be excluded that other organs may be used as starting materials for the isolation of the virus of the invention.

The Newcastle disease virus of the present invention may be in live, live attenuated or inactivated form.

In another aspect, the present invention provides a vaccine for protecting poultry against diseases leading to mortality, with clinical manifestations of depression, green diarrhea, speculum, conjunctivitis, etc. caused by Newcastle disease virus infection. Newcastle disease virus and a pharmaceutically acceptable carrier or diluent.

Newcastle disease virus of the invention can be included in the vaccine as a live, attenuated or inactivated form. If the Newcastle disease virus is in live, attenuated or inactivated form, the nature of the avian Newcastle disease virus inducing the above-mentioned diseases is significantly reduced or completely eliminated.

The weakening of the Newcastle disease virus of the present invention can be achieved by methods well known in the art for this purpose, for example as disclosed in Gouvea et al. (Virology 126, 240-247, 1983). Briefly, virus isolates are inoculated into chicken embryo fibroblast cells (CEF) in primary chickens after virus isolation from the target animal. If the isolate is unable to produce a cytopathic effect (CPE), the virus is inoculated repeatedly (eg 3-10 times) until CPE is observed. As soon as CPE is visible, cells and cell cultures are collected, frozen and thawed, cleared by centrifugation, and the supernatants containing Newcastle disease virus isolates are split and stored at -20 ° C. This process can be repeated (eg 10-100 times) to further weaken the virus.

Vaccines of the invention can be prepared according to conventional methods commonly used, for example, in commercial live inactivated Newcastle disease virus vaccines. Preparation of veterinary vaccine compositions is particularly described in "Handbunch der Schutzimpfungen in der Tiermedizin" (ed. Mays, A. et al., Verlag Paul Parey, Berlin und Hamburg, Germany, 1984) and "Vaccines or Veterinary Applications" (ed. Peters, AR). Et al., Butterworth-Heinemann Ltd, 1993.

In brief, the susceptible substrate is inoculated with the Newcastle disease virus of the present invention in a live or live attenuated form, and propagated until the virus replicates at a predetermined infectivity or antigen content (mass). The reovirus bearing the substance can then be harvested and formulated into a pharmaceutical composition with prophylactic activity.

All substrates that can be used for replication of the Newcastle Disease Virus as defined above can be used to generate a vaccine according to the present invention if necessary after the Newcastle Disease Virus has adapted to the substrate.

Suitable substrates include primary (algal) cell cultures (e.g., chicken embryo liver cells (CEL), chicken embryo fibroblast cells (CEF), or chicken kidney cells (chicken). kidney cells (CK)), mammalian cell lines (eg Vero cell lines) or avian cell lines such as BGN-70 cell lines, QT-35, QM-7 or LMH. Usually, after inoculation of the virus the virus is propagated for 3-10 days, after which the cell culture supernatant is collected and, if necessary, filtered or centrifuged to remove cell debris.

Alternatively, after the Newcastle disease virus of the present invention is propagated in embryonated eggs, Newcastle disease virus material may be harvested by conventional methods.

Vaccines of the invention containing live attenuated virus are prepared and marketed in the form of lyophilized or (frozen) suspensions. The vaccine further comprises a diluent or pharmaceutically acceptable carrier commonly used in the composition. Carriers include stabilizers, preservatives and buffers. Suitable stabilizers are, for example, SPGA, hydrocarbons (eg sorbitol, mannitol, starch, sucrose, dextran, glutamate, or glucose), proteins (eg dried milk serum, albumin or casein) or degradation products thereof. . Suitable buffers are for example alkali metal phosphates. Suitable preservatives are thimerosal, methiolate, gentamicin. Dilutions are water, aqueous buffers (eg buffered saline), alcohols and polyols (eg glycerol).

If necessary, the live vaccine of the present invention may contain an adjuvant. Examples of suitable compounds and compositions with adjuvant activity are the same as mentioned below for the preparation of inactivated vaccines.

The live vaccine of the present invention can be administered, for example, by intramuscular or subcutaneous injection, but the live vaccine is preferably administered by a low cost mass dose technique commonly used for Newcastle disease virus vaccination. This technique includes drinking and spray vaccination.

Another method for the administration of live vaccines includes in ovo administration, eye drop administration and beak dipping administration.

The present invention also provides a vaccine comprising the Newcastle disease virus in an inactivated form for Newcastle disease. The main advantage of inactivated vaccines is that long levels of high levels of protective antibodies persist. This property makes inactivated vaccines particularly suitable for breeder vaccination.

The purpose of inactivating the harvested virus after the propagation step is to prevent regeneration of the virus. In general, this can be done by chemical or physical means. Chemical inactivation can be performed, for example, by treating the virus with enzymes, formaldehyde, β-propiolactone, ethylene-imine or derivatives thereof. If necessary, the deactivated compound is later neutralized. Formaldehyde inactivated material may be neutralized with, for example, thiosulfate. Physical inactivation may preferably be carried out by irradiating the virus with sufficient energy, for example UV light. If necessary, the pH may be adjusted to a value of about 7 after treatment.

Vaccines containing the inactivated Newcastle disease virus may comprise one or more of the aforementioned pharmaceutically acceptable carriers or diluents suitable for this purpose, for example. Preferably, the inactivated vaccine of the present invention comprises one or more compounds with adjuvant activity. Suitable compounds or compositions for this purpose include vegetable oils such as aluminum hydroxide, -phosphate, -oxide, mineral oil, vitamin E acetate, and oil-in-water or water-in-oil emulsions based on saponins.

Inactivated vaccines are usually administered parenterally, ie intramuscularly or subcutaneously.

The vaccine of the present invention comprises, as an active ingredient, an effective amount of Newcastle disease virus, that is, an amount that immunizes Newcastle disease virus material that will induce immunity in vaccinated birds or their offspring against attack by toxic viruses (antigen administration). Immunity is defined herein to induce a significantly higher level of protection in avian populations after vaccination compared to the non-vaccinated group.

Typically, the live vaccine of the present invention can be administered at a dose of 10 ² -10 ⁹ TCID ₅₀ , preferably 10 ² -10 ⁶ TCID ₅₀ per bird. Inactivated vaccines also contain antigens equivalent to 10 ⁴ -10 ¹⁰ TCID ₅₀ per bird.

The Newcastle disease virus vaccine of the present invention can be used effectively in chickens, but other poultry such as turkey, guinea fowl and quail can also be vaccinated effectively. Chickens include edible chickens, cloned cattle, and laying eggs.

The age of the animal receiving the live or inactivated vaccine according to the present invention is the same as the age of the animal receiving the live- or inactivated Newcastle disease virus vaccine currently on the market. For example, edible chickens are directly vaccinated with the live weakened vaccine of the present invention from day 1 of birth. Vaccination of parental livestock, such as breeders of edible chickens, can be performed with the live, attenuated or inactivated vaccine of the present invention, or a combination of both. The advantages of this type of vaccine program include the direct protection of one day old offspring provided by antibodies from the maternal line that are delivered vertically to the offspring. A typical breeder's vaccination program involves inoculating a six week old breeder with a live, weakened vaccine followed by an inactivated vaccine at 14-18 weeks of age. Alternatively, live vaccines may be given two doses with inactivated vaccines at 10-12 weeks of age and 16-18 weeks of age.

The present invention also encompasses vaccine combinations comprising one or more vaccine components of other pathogens infected with poultry in addition to the Newcastle disease virus of the present invention.

Preferably the vaccine component in the vaccine combination is a live, attenuated or inactivated form of the pathogen infected with the poultry.

In particular, the present invention provides a vaccine combination in which all vaccine components are in an inactivated form.

Vaccine combinations include infectious bronchitis virus (IBV), avian reovirus, Infectious bursal disease virus (IBDV), fowl adenovirus (FAdV), and hypogonadal virus (egg). It is preferable to include one or more (inactivated) vaccine strains of drop syndrome 76 'virus, EDS 76' virus and turkey rhinotracheitis virus (TRTV).

In the present invention, it is possible to effectively protect poultry from Newcastle disease by providing a new Newcastle disease virus, a vaccine comprising the same, and a method for producing the same.

Hereinafter, the present invention will be described in more detail with reference to non-limiting examples. However, the following examples are illustrated to illustrate the present invention and the scope of the present invention is not to be construed as limited by the following examples.

Example 1 Isolation of Newcastle Disease Virus K148 / 08 strain from Mallard duck

1) Isolation and Identification of Newcastle Disease Virus

In 2008, after collecting fecals from Mallard duck, diluted to 10% weight / volume (w / v) in PBS and centrifuged, the supernatant was filtered to 0.45㎛ pore size to exclude bacterial contamination. Specific pathogen free (SPF) was inoculated in chicken embryos and cultured in a 37 ℃ thermostat. After incubation for 3 days, allantoic fluid was collected from chicken eggs and reacted with 10% SPF chicken erythrocytes in a 1: 1 ratio to determine the hemagglutination activity of HA. It was. Fecal duck feces by performing polymerase chain reaction (PCR) on the matrix (M) protein gene, a specific protein of Newcastle disease virus (NDV), against hemagglutinin virus It was confirmed that the Newcastle disease virus was present within.

2) Test for other pathogens in the live seed vaccine Master Seed

S (Spike) of M protein gene of Avian influenza virus (AIV), Infectious bronchitis virus (IBBV) PCR was performed on protein genes, and it was confirmed that there were no pathogens other than Newcastle disease virus in wild duck feces.

3) Production of live vaccine Seed virus stock

Urinary fluid obtained from wild-capped poultry eggs was prepared as a master seed, and two seeded virus stocks were prepared. The virus content of Seed virus stock was found to be 10 ^9.0 EID ₅₀ / ml as a result of calculating 50% Egg infectious dose (EID ₅₀ ).

4) Analysis of the nucleotide sequence of the F protein gene of Newcastle disease virus K148 / 08 strain

In order to analyze the F protein nucleotide sequence of Newcastle disease virus isolated from wild duck, PCR was performed using Primer which can amplify the 508th sequence of the F protein gene from the 1055th sequence of the M protein gene. Was carried out.

Example 2: Pathogenicity Test of NDV K148 / 08 Isolates

1) Pathogenicity Identification through NDV Fusion (F) Gene Sequencing

NDV exhibits pathogenicity as the NDV surface protein F protein is cleaved by enzymes in vivo, and the type of enzyme cleaved according to the Motif sequence of amino acids 111-117 of the F protein determines the pathogenicity. Difference appears. The amino acid sequence of ¹¹² G / EK / RQG / ER ¹¹⁶ -L ¹¹⁷ appears at the cleavage site of the F protein in NDV attenuated or non-pathogenic strains, whereas the amino acid sequence of ¹¹² R / K- RQK / RR ¹¹⁶ -F ¹¹⁷ is found in NDV toxicists. Appears.

In order to identify the pathogenic classification of wild duck-derived NDV K148 / 08 isolates, the sequence of amino acids 111-117 of the F protein was analyzed. As a result, NDV K148 / 08 isolates were classified as non-pathogenic because they had a characteristic of ¹¹² GKQGRL- ¹¹⁷ at the cleavage site of F protein. (Figure 2)

2) Pathogenicity Identification Using 10-day-old SPF Gyetae-Aran and 1-day-old SPF Superspine

In addition to F gene sequencing, NDV pathogenicity measures include mean death time (MDT) measurements using SPF embryonic eggs and intracerebral pathogenic index (ICPI) measurements using 1-day-old SPF herbivores. have. In case of NDV poisoning, ICPI scores are within 1 ~ 2 points and MDT within 60 hours.

Diluted 10-fold dilution of NDV K148 / 08 isolates into the cerebrum of 1-day-old SPF herbivore and observed for 8 days of clinical symptoms and mortality. 0 points for normal, 1 point for clinical symptoms, and 2 points for mortality. The cerebral pathogenicity index was calculated by summing up and dividing by total inoculation sheath harvest. As a result, the cerebral pathogenicity index of NDV K148 / 08 isolate was measured as 0.1.

In addition, the K148 / 08 isolates were diluted 10 ^-6 to 10 ^-9 and then inoculated in 10-day-old SPF chick embryos for 7 days, causing mortality of the maximum dilution factor that causes mortality in all inoculated chick embryos. The mean mortality time was tested by measuring the time. The average mortality of NDV K148 / 08 isolates was 144 hours.

Based on the cerebral pathogenicity index and mean mortality time, we identified NDV K148 / 08 isolate as non-pathogenic heat resistant strain. (Figure 2)

Example  3: NDV K148 / 08 heat resistance test of separationism

Some of the commercially available NDV vaccine strains (Fig. 1) are characterized by heat resistance (Thermostability) which does not lower the titer of virus infection even after exposure to 56 ° C for more than 1 hour, which is a cold-chain during storage and transportation of the vaccine. There is an advantage that does not need). Therefore, the virus titer (50% egg infectious dose, 50% Egg infectious dose, EID ₅₀ ) and hemagglutination capacity after treatment for 1 hour at 56 ° C were tested to test the heat resistance of NDV K148 / 08 isolates. Heat resistance was tested by comparing the hemagglutination activity (HA). As a result, NDV K148 / 08 isolate had heat resistance because virus titer and hemagglutination ability did not decrease even after treatment at 56 ℃ for 1 hour. (Fig. 3)

Example  4: SPF From chicken NDV K148 / 08 Immunogenicity and Protective Efficacy Testing of Isolates

NDV is known that humoral immunity during vaccination plays an important role in disease defense. Therefore, hemagglutination inhibition (HI) titer using the characteristics of NDV that aggregates chicken red blood cells has been used as a useful method for evaluating the efficacy of the vaccine. In addition, the NDV National Vaccine Screening Standard (National Veterinary Quarantine Service) indicates that the NDV vaccination rate is 80% or higher when the NDV vaccination vaccination is administered. have.

In order to confirm the neutralizing antibody formation ability and the protective ability against NDV prevalent in NDV K148 / 08 isolates, three 120-week-old SPF chickens were divided into three groups and two NDVs commercialized with NDV K148 / 08 isolates. Vaccines (Avinew, Merial, France / DSB-HP, Daesung, Korea) were inoculated with a dose of 10 ^5.0 EID ₅₀ . Two weeks after vaccination, hemagglutination was performed with serum collected to measure HI antibody titer. HI dilutes serum with PBS in V-shape bottom 96 well microplate (Greiner bio-one, Kremsmuenster, Austria), mixes equal amounts (25 μl) of NDV antigen in 4HA, and sensitizes at room temperature for 30 minutes. 25 μl of 1% chicken erythrocytes were added and mixed well, and after reading for 40 minutes at room temperature. HI titer was the dilution factor of the final well where hemagglutination occurred.

In order to evaluate the protection against NDV poisonous strains of NDV K148 / 08 isolates, the clinical symptoms after challenge with the genotype NDV strains (Kr-005 / 00), which are currently in prevalence in Korea and Asia, 2 weeks after vaccination, Mortality was observed to calculate defensive capacity. As a result, the NDV K148 / 08 isolate vaccinated group showed higher antibody titers than the two commercially available NDV vaccines. It was confirmed that it can be suppressed. (Figure 4)

Example 5: Safety test in 1-day-old broiler herbivore with vaccinated NDV K148 / 08 isolate

To assess the safety of NDV K148 / 08 isolates, one-day-old broiler herbivore was divided into three groups, and NDV K148 / 08 isolates and two commercialized vaccines (Avinew, Merial, France / DSB-HP, Daesung, Korea) Was sprayed using a Desvac sprayer (Intervett, The Netherlands) of 115 탆 spray particle size, which is the most used in Korea, at a dose of 10 ^5.0 EID ₅₀ per allowance.

Four, seven, eleven, and 14 days after vaccination, the upper, middle, and lower organs were taken from three test groups for each group, and the cross section of the tracheal ring was observed under a microscope to evaluate the degree of cilia. 0 points if cilia are active throughout the cross section of the engine ring, 1 point if 25% of the cilia are stopped, 2 points if the 50% of the cilia are stopped, 3 points if the cilia are stopped, and 3 points of the whole ring. If it was stopped, the ciliary loss factor was calculated by calculating the mean value as 4 points. In addition, 4, 7, 11, and 14 days after vaccination, upper, middle, and lower organs and lung tissues were collected from three test groups in each group. Tissue sections were prepared and tissue lesions were observed by staining with Hematoxylin & Eosin. The tissue lesion index was regarded as 0 points for normal tissue, 1 point for localized tissue lesions, 2 points for localized lesions, and 3 points evenly observed throughout the site. Finally, after weighing 20 animals in each group 4 weeks after vaccination, the average value was calculated to observe the effect of vaccination on chicken growth rate.

The test results showed that the ciliary motor loss index of the test system vaccinated with the NDV K148 / 08 isolate was lower than the safety criteria (2.0) of the European Pharmacopoeia, and the same or lower than that of the commercial vaccine. In histological lesions, no significant histological lesions were observed in both the NDV K148 / 08 isolate and two commercial vaccination groups. In addition, the increase rate of NDV K148 / 08 isolates was significantly higher than that of commercial vaccinated groups, indicating that NDV K148 / 08 isolates were safe vaccines with less effect on gain ratio than commercialized vaccines. (See FIGS. 5, 6 and 7)

Example 6: Safety test in 1-day-old broiler herbivore when inoculated with fine particles of NDV K148 / 08 isolates

To evaluate the safety and efficacy of injecting fine spray particles (50 µm) against NDV K148 / 08 isolates that have demonstrated efficacy and safety as a vaccine for spray vaccination using a conventional spray inoculator (Desvac, particle size 115 µm) 1 Separate days old broiler sod weight into three groups and NDV K148 / 08 isolates and commercialize two NDV baeksinju (Avinew, Merial, France / DSB-HP, Daesung, Korea), the allowance 10 ^5.0 EID 50 to ₅₀ used in the country The fine spray was inoculated using a cabinet-type sprayer (SK-MO-Auto-3000, Three Shine, Korea) having a particle size of spray particles.

After 4, 7, 11, and 14 days of vaccination, the upper, middle, and lower organs were collected from three test groups in each group, and then examined for cilia movement using a microscope. If 25% of the cilia are stopped, 1 point, 50% of the cilia are stopped, 2 points, 75% of the cilia are stopped, 3 points; The ciliary dissipation index was calculated. In addition, 4, 7, 11, and 14 days after vaccination, the upper, middle, and lower organs and lung tissues were collected from three test groups, and tissue sections were prepared and tissue lesions were observed by staining with Hematoxylin & Eosin. Tissue lesion index was considered to be 0 for normal tissue, 1 for localized lesions, 2 for multiple lesions, and 3 for evenly observed lesions. Four weeks after vaccination, the weight of 20 test subjects was measured in each group, and the average value was calculated to observe the effect of the vaccine on the weight gain rate.

The results showed that cilia loss index was found to be higher than the European Pharmacopoeia safety standard (2.0) in the test system in which the NDV vaccine was commercially vaccinated, whereas the cilia loss index in the NDV K148 / 08 isolate was fine. Observations at levels below safety criteria confirmed that NDV K148 / 08 isolates had less effect on respiratory cilia than commercial NDV vaccine strains. In addition, the results of histological examination after fine spraying showed that ciliary loss, lymphocytic infiltration, mucosal congestion, etc. were observed between 4 and 7 days of vaccination in both the NDV K148 / 08 isolates and the commercially available two vaccine strains. Respiratory tissue damage was observed, but the histological lesion index of the NDV K148 / 08 isolates was the same or lower than that of the two vaccinated strains. Tissue lesions were observed up to 11 days after inoculation, whereas tissue lesions were observed up to 7 days after vaccination in the NDV K148 / 08 isolate vaccination system. It was found to be less than in the vaccine. In the test of the increase rate after spray vaccination, the test system spray-inoculated with NDV K148 / 08 isolate showed significantly higher gain rate than the test system spray-inoculated with two commercialized vaccines. The chief was confirmed. (FIGS. 8-10)

1 is a table listing domestically sold ND live vaccine vaccine.

Figure 2 is a table showing the results of pathogenic identification using NDV F protein amino acid sequence analysis and SPF system embryos and SPF herbivore.

3 is a table showing the results of virus titer and hemagglutination change after 56 ℃ heat treatment of NDV K148 / 08 isolate.

 Figure 4 is a table showing the results of defense test against HI antibody formation ability and NDV virulent challenge after inoculating the NDV K148 / 08 isolate in the 1-day-old SPF herbivores.

Figure 5 is a table comparing the weight gain after 4 weeks of NDV K148 / 08 spray vaccination.

6 is a graph showing the results of evaluating the ciliary motor loss index after spray vaccination of K-148 NDV isolate.

7 is a table showing the results of evaluating the respiratory tract tissue lesion index after the spray vaccine vaccination of NDV K148 / 08 isolate.

FIG. 8 is a table comparing weight gain after 4 weeks of NDV K148 / 08 isolate injection.

9 is a graph showing the results of evaluating the ciliary motor loss index after spraying the fine particle of the K-148 NDV isolate.

10 is a table showing the results of evaluating the respiratory tract tissue lesion index after the injection of the NDV K148 / 08 isolated fine particle spray vaccine.

Claims (11)

NDV K148 / 08 Newcastle Disease Virus deposited with Accession Number KCTC 11570BP. The Newcastle disease virus according to claim 1, wherein the virus is isolated from domestic wild ducks. A vaccine for poultry protection against diseases caused from Newcastle disease virus infection, comprising the Newcastle disease virus of claim 1 or 2 and a pharmaceutically acceptable carrier or diluent. 4. The vaccine of claim 3 wherein said Newcastle disease virus is in live, attenuated form. The vaccine of claim 3 wherein said Newcastle disease virus is in an inactivated form. 4. The vaccine of claim 3 wherein said vaccine further comprises an adjuvant. The vaccine of claim 3, wherein the vaccine further comprises one or more vaccine components of other pathogens infectious to poultry. a) inoculating a susceptible substrate with the Newcastle disease virus as defined in claim 1; b) propagating the Newcastle disease virus; And c) collecting the Newcastle disease virus-containing material. The method of claim 8, wherein the substrate is one cell selected from the group consisting of embryonic liver cells (CEL), chicken embryo fibroblasts (CEF), chicken kidney cells (CK), and Vero cell line. Method of producing Newcastle disease virus, characterized in that. 10. A method of inducing a Newcastle disease virus infection comprising the step of combining the collected Newcastle disease virus obtained by the method of claim 8 with a pharmaceutically acceptable carrier or diluent, if necessary, after the inactivation of the Newcastle disease virus. Method for preparing a vaccine for poultry protection against diseases. A method of controlling a disease resulting from Newcastle disease infection in poultry comprising administering the vaccine of claim 3 to birds.

Images