Characterization of Newcastle Disease Viruses Isolated from Chickens and Ducks in Tamilnadu,India

During 1993, outbreaks of Newcastle disease occurred on many farms in Tamilnadu, India. Six Newcastle disease virus (NDV) isolates were obtained from the chickens on five different farms and from the birds on one duck farm during outbreaks of the disease. All the isolates were characterized as velogenic, based on the mean death time, intravenous pathogenicity index, intracerebral pathogenicity index (ICPI), stability of haemagglutinin at 56°C, agglutination of equine erythrocytes, haemagglutination elution pattern and adsorption of haemagglutinin by chick brain cells. The isolate obtained from ducks resembled a group D strain, based on its ICPI and its reaction with a panel of monoclonal antibodies. The other five NDV isolates obtained from chickens were placed in groups B(1), C1(2) and D(2) on the basis of their binding patterns with the panel of monoclonal antibodies. In challenge experiments, it was found that LaSota vaccine provided 100% protection against each of these field isolates and against a local NDV strain obtained from the Institute of Veterinary Preventive Medicine, Tamilnadu, India, while unvaccinated chickens succumbed to challenge. The possible origin of epizootic viruses causing outbreaks in vaccinated flocks is discussed.     

An investigation of the disease status of village poultry in Mauritania

Three contagious poultry diseases in Mauritania were studied. Serum samples and tracheal swabs were taken from 80 chickens in village poultry flocks in each of three different regions. Antibodies against Newcastle disease virus (NDV) were detected in 4.8% of chickens, antibodies against Gumboro disease virus in 15.8% of chickens and antibodies against Salmonella pullorum in 6.2% of chickens. Six isolates of NDV were made, of which four formed plaques on chicken embryo fibroblast monolayers. Seropositivity was highest in the Trarza region, bordering the River Senegal, and four out of six of the NDV isolates were made from chickens in this region.     

鸵鸟新城疫病毒野毒株的分离及其生物学特性

用鸡胚从疑似鸵鸟新城疫的送检病料中分离到 2株病毒。 2株病毒均能凝集鸡红细胞 ,且能被抗新城疫病毒 ( NDV)阳性血清抑制 ;用抗 NDV单抗 PEG夹心 ELISA测定 2株分离毒均为阳性。对其中 1株作进一步生物学特性鉴定 ,按照国际上规定的 NDV毒力判定标准测定的该毒株最低致死量致死鸡胚的平均死亡时间 ( MDT)为 54h,1日龄雏鸡脑内接种致病指数 ( ICPI)为 1 .88,6周龄雏鸡静脉接种致病指数 ( IVPI)为2 .75。结果表明 ,本分离株为鸵鸟新城疫病毒强毒     

Paramyxovirus type 1 infections of racing pigeons: 1 characterisation of isolated viruses

Viruses isolated from field outbreaks of disease in racing pigeons in continental Europe and Great Britain were shown to be identical by serological tests using conventional chicken antisera and mouse monoclonal antibodies. The pigeon viruses showed high levels of cross-reaction to Newcastle disease virus (NDV) in haemagglutination inhibition tests and Madin-Darby bovine kidney cells infected with pigeon virus isolates bound three out of nine mouse monoclonal antibodies prepared against NDV Ulster 2C. These results confirm their classification in the paramyxovirus type 1 serotype of avian paramyxoviruses. However, the pigeon viruses could be distinguished from more classical paramyxovirus type 1 viruses by the significantly different titres obtained in haemagglutination inhibition tests, the failure of mouse monoclonal antibodies directed against the HN1 epitope of NDV Ulster 2C to inhibit their haemagglutinating activity and a unique binding pattern seen with the nine mouse monoclonal antibodies.     

Antigenic heterogeneity amongst the field isolates of Newcastle Disease Virus (NDV) in relation to the vaccine strain. Part II: studies on viruses isolated from domestic birds in Israel

Forty three Newcastle disease virus (NDV) strains isolated before and during 1997 in Israel from domestic birds were studied by means of the three panels of monoclonal antibodies prepared against all the viral envelope proteins in order to reveal the possible antigenic differences between them and the VH strain used in Israel for poultry vaccination. Three isolates were found to have significant antigenic differences in the hemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins as compared to the vaccine strain. As to the matrix protein, almost all the viruses isolated during the year 1997 were found to have considerable differences from the vaccine strain in two of four antigenic sites.     

Characterization of a battery of monoclonal antibodies for differentiation of Newcastle disease virus and pigeon paramyxovirus-1 strains

Twenty monoclonal antibodies (MCAs) prepared against the velogenic GB-Texas strain of Newcastle disease virus (NDV) and the type 1 pigeon paramyxovirus (PPMV-1) were characterized and examined as potential immunodiagnostic reagents. All MCAs generated were found to bind specifically, but with varying reactivity, to various NDV strains in direct binding assays. In addition, MCA 15C4 neutralized and inhibited hemagglutination (HA) of all lentogenic, mesogenic, and velogenic NDV strains tested but not the PPMV-1 strain. Antibody 10D11 also inhibited HA activity, but inhibition was more selective and limited to the mesogenic and domestic or indigenous velogenic strains of NDV. MCA 79 reacted in all serologic assays with an antigenic site common to all serotype 1 avian paramyxoviruses. Passive immunization studies involving three different neutralizing MCAs (35, 79, and 15C4) showed that enhanced, but not complete, protection against virulent NDV challenge was provided when the three MCAs were administered in combination.     

新城疫La Sota疫苗对山东分离株的免疫保护试验

用标准疫苗株La Sota活疫苗在隔离器中接种SPF鸡,进行免疫保护试验.免疫后,每7 d采血监测NDV抗体,免疫后2周,利用经过鉴定的新城疫病毒(NDV)潍坊毒SGM01、昌乐毒SCL03、东营毒HY、日照毒SRZ03、莘县毒SSX03和标准强度F48E9分别进行攻毒试验,同时设SPF鸡对照.每天观察,及时剖检发病鸡,检查鸡群病变,确定疫苗的保护性.结果表明,La Sota疫苗能对除SGM01和HY以外的病毒攻击的SPF鸡提供较好的保护.     

A reservoir of virulent Newcastle disease virus in village chicken flocks

Village chicken flocks in 6 different regions of Morocco were studied for the presence of Newcastle disease. Three of the regions contained industrialised poultry farming and 3 were isolated mountainous regions with no industrialised poultry farming. Serum samples and tracheal swabs were taken from 100 chickens in each region. Antibodies against Newcastle disease virus (NDV) were found in chickens from every region. Forty-one isolates of NDV were obtained including some from chickens in every region. Two virus isolates from each region characterised and all were found to be velogenic. Thus, village chicken flocks throughout Morocco harbour a reservoir of virulent NDV, independently of industrialised farms.     

从自然发病鸡中分离出一株新城疫病毒

应用电镜技术、血清学检查、生物学试验等方法,从未注射过疫苗的某自然发病鸡群分离出一株新城疫病毒。该分离株对１０日龄鸡胚的平均致死时间为１１６小时,静脉接种致病指数为０．０６,其毒力介于新城疫病毒的ＬａＳｏｔａ株与Ｖ４株之间。免疫原性试验结果表明,该分离株具有免疫后无不良反应、免疫后６天开始产生抗体、产生的抗体效价高、对新城疫强毒的攻击能１００％保护等特点,是一株良好的新城疫候选疫苗株     

Comparative characteristics of the Israeli Newcastle disease virus field strains by means of a wide panel of monoclonal antibodies against hemagglutinin-neuraminidase glycoprotein

Fourteen mouse monoclonal antibodies (MAB) were tested for their ability to react with 15 reference and 52 local Newcastle disease virus (NDV) strains isolated in Israel during the last decade from feral birds. All the field isolates had no antigenic difference when examined by classic serological tests. However, MAB-mediated analysis revealed wide antigenic heterogeneity amongst the studied viruses. By the pattern of the MAB reactivity, all the isolates could be distributed into 13 groups.     

Pathogenicity and cross-protection of pigeon paramyxovirus-1 and Newcastle disease virus in young chickens

Avian paramyxovirus-1 (PMV-1) isolates from Delaware racing pigeons were compared with Newcastle disease virus (NDV) in pathogenicity and cross-protection studies in young chickens. The pathogenicity of pigeon PMV-1 isolates was more closely related to mesogenic (Roakin) NDV than to lentogenic (La Sota) or velogenic (Texas GB) NDV strains. Pigeon PMV-1 produced 100% mortality in 1-day-old NDV-susceptible chickens following intratracheal and intracerebral inoculation. Laboratory tests often used in conjunction with chicken pathogenicity procedures for patho-typing NDV gave conflicting results. Pigeon PMV-1 isolates produced large clear plaques (up to 3.5 mm) in chicken-embryo-fibroblast cultures. Chicken embryo mean death times were considerably greater for pigeon PMV-1 (88 and 109 hr) than for Roakin (66 hr) and Texas GB (48 hr). B1 strain NDV and pigeon PMV-1 produced complete cross-protection in challenge studies in chickens. Extensive cross-reaction between pigeon PMV-1 and NDV occurred in hemagglutination-inhibition tests using polyclonal antisera. However, pigeon PMV-1 and NDV were readily distinguishable using a NDV monoclonal antibody, 2F12.     

Protective antibody response produced by the chickens vaccinated with green coloured thermostable Newcastle disease virus

The efficacy of green-coloured (GC) I-2 Newcastle disease vaccine was determined in the present study. I-2 vaccine was mixed with a green coloured dye and stored at 4°C for 6 months while assayed for the virus infectivity at a monthly interval. Chickens were vaccinated with the GC vaccine by eye drop. Serum samples were collected from all birds before and after vaccination at weekly interval for 4 weeks and tested for haemagglutination-inhibition (HI) antibody against Newcastle disease virus (NDV). These chickens were challenged with NDV virulent strain four weeks after vaccination. The results showed that there was no difference between the infectivity titres of GC and uncoloured vaccines. However, chickens vaccinated with GC vaccine produced higher HI antibody titres than chickens vaccinated with uncoloured vaccine. Results from the challenge trial showed that all vaccinated chickens survived whereas all unvaccinated chickens died. The findings from this study have shown that the GC vaccine is safe and produced protective antibodies against NDV in vaccinated chickens. Wambura, P. N., 2008. Protective antibody response produced by the chickens vaccinated with green coloured thermostable Newcastle disease virus. Tropical Animal Health and Production.     

Identification of a variable epitope on the Newcastle disease virus hemagglutinin–neuraminidase protein

Fifteen virulent Newcastle disease viruses (NDVs) were isolated from diseased birds in Eastern China in 2005. To investigate the antigenic variation in the epitopes on NDV hemagglutinin–neuraminidase (HN) protein, these isolates, together with six reference strains, were subjected to the hemagglutination inhibition (HI) tests using five HI-positive monoclonal antibodies (MAbs) against velogenic NDV strain ZJ1. The MAbs 2G5, 3A4, 3B5 and 6B1 recognized 12 of the 15 NDV isolates, and exhibited HI activity towards the six reference strains. However, these MAbs did not react with three local isolates, JS-02/05, JS-06/05 and JS-10/05. HN gene sequence analysis of all NDV strains revealed that these MAb-resistant NDV isolates possessed residue K at position 347 of the HN protein, whereas all remaining strains possessed E or G at the same site. To determine the contribution of the residue at position 347 to antigenic epitope formation, we generated by reverse genetics two recombinant viruses, ZJ1HNK with an E347K mutation on ZJ1 HN, and JSHNE with a K347E mutation on JS-06/05 HN. The HI test demonstrated that ZJ1HNK lost reactivity with MAbs 2G5, 3A4, 3B5 and 6B1, whereas JSHNE did react with these MAbs. Further verification by immunofluorescent assay demonstrated that residue 347 was a critical determinant for formation of the antigenic epitope (residues 345–353) on the HN protein.     

Two types of Newcastle disease viruses isolated from feral birds in Western Australia detected by monoclonal antibodies

Thirteen viruses isolated from feral birds and one isolated from a domestic duck, obtained in 1979-1980 during a survey of birds in Western Australia, were shown to be Newcastle disease viruses of low virulence for chickens. The binding of mouse monoclonal antibodies, raised against NDV-Ulster 2C to MDBK cells infected with the isolates was assessed using an indirect immunoperoxidase test. Five viruses caused binding of all 9 monoclonal antibodies tested, whereas the other 9 isolates induced binding of only 4 monoclonal antibodies.     

Detection of antibody-forming cells directed against Newcastle disease virus and their immunoglobulin class by double immunoenzyme histochemistry

Antibody-forming cells (AFCs) against Newcastle disease virus (NDV) and their immunoglobulin (Ig) class were demonstrated by a double immuno-enzyme histochemical technique. The AFCs were stained and quantified in spleen sections of chickens euthanatized at day 7 postexposure to the Roakin strain of NDV. The sections were incubated with NDV to determine the specificity of the AFCs. Bound virus was subsequently visualized with a primary monoclonal antibody (MAb), a secondary horseradish peroxidase-conjugated MAb, and 3-amino-9-ethylcarbazole as substrate. IgM and IgA were stained with MAbs and an alkaline phosphatase (AP)-conjugated secondary antibody. IgG class antibodies were demonstrated with an AP-conjugated rabbit serum. The final substrate for the three Igs was naphthol AS-MX-phosphate and fast blue BB. About 64-159/mm2 AFCs against NDV were detected. Of these virus-binding cells, about 55% produced IgM, 37% produced IgG, and the remaining 8% produced IgA.     

THE RESPONSE OF AUSTRALIAN CHICKENS NATURALLY INFECTED WITH AVIRULENT NEWCASTLE DISEASE VIRUS TO CHALLENGE WITH VELOGENIC NEWCASTLE DISEASE VIRUS

SUMMARY Sixty-eight breeder chickens, 4 to 12 months of age, were taken from Australian flocks that had been naturally infected with avirulent Newcastle disease virus (NDV) and transported by air to Malaysia. Nearly all the breeders had haemagglutination inhibition antibodies to NDV, at titres of from 2 to 128. Thirty-two were inoculated intranasally with an Asian, velogenic, viscerotropic strain of NDV and all survived this challenge. Thirty-six were exposed to contact infection with the same velogenic NDV and 2 died of Newcastle disease within 14 days. The levels of haemagglutination inhibition antibodies against NDV increased in the surviving breeders after challenge, reaching 2048 or greater in a few birds. Velogenic NDV was isolated from a cloacal swab from one clinically normal breeder 10 days after challenge by contact. Cloacal swabs taken 7 to 10 days after challenge from another 23 breeders yielded no NDV. Twenty-four broilers, 7 weeks of age, were also transported from Australia to Malaysia. All lacked detectable haemagglutination inhibition antibody to NDV and they were from a flock with no detectable antibody to NDV. Twelve were challenged with velogenic NDV intranasally and 12 were subjected to contact challenge. All broilers died of Newcastle disease within 13 days.     

新城疫病毒单克隆抗体的制备及与不同分离株的反应性

以纯化的新城疫病毒(Newcastle dis-ease virus,NDV)弱毒株La Sota为免疫原,接种6周龄~8周龄Balb/c小鼠。最后一次加强免疫后3 d,取小鼠脾细胞与骨髓瘤细胞(SP2/0)融合。以纯化的NDV建立了筛选单抗的间接ELISA,经多次检测,3次亚克隆,共获得5株NDV特异性单抗,这5株单抗与NDV不同分离株的反应性存在差异,表明部分NDV毒株抗原性发生了变异。     

INTERACTION BETWEEN INFECTIOUS BURSAL DISEASE VIRUS AND NEWCASTLE DISEASE VIRUS IN CHICKENS

The Australian strain of infectious bursal disease virus (IBDV), 002/73, affected the response of chickens to Newcastle disease virus (NDV). The titre of serum antibodies to NDV in chickens infected with IBDV was significantly lower than that of birds infected with NDV alone. It also appeared that IBDV affected NDV excretion from chickens as NDV was more frequently isolated from chickens infected with IBDV, IBDV infection did not alter the pathogenicity of NDV in chickens. This Australian strain of IBDV therefore appeared to be immunodepressive in one-day-old chickens.     

Selection of thermostable Newcastle disease virus progeny from reference and vaccine strains.

In a study of low-virulence Newcastle disease virus (NDV) isolates from poultry, 38% of the isolates had a more thermostable hemagglutinin than the lentogenic reference strains B1 and La Sota or live vaccines derived from those strains. Whether those strains with a more thermostable hemagglutinin are truly indigenous or whether they could have originated from vaccines used in the flocks was unknown. Seven monovalent NDV vaccines of B1 or La Sota type and reference B1 and La Sota strains were heat treated at 56 C to select variants more thermostable than the parent virus. Four thermal treatment cycles were completed, and virus propagated from the second and fourth heat treatments was assayed for changes in thermostability and antigenicity. The hemagglutinin thermostability of all vaccine and reference strain variants increased from the initial < or =10 min to > or =120 min after four treatments. Antigenic changes evaluated by hemagglutination inhibition against NDV monoclonal antibodies identified changes in only the heat-treated La Sota strains. The results demonstrate that the field isolates with a more thermostable hemagglutinin could have been derived by selection from the heterogenous NDV populations in vaccine strains and that minor antigenic changes may be a result of that selection.     

Vaccination against Newcastle disease with a recombinant baculovirus hemagglutinin-neuraminidase subunit vaccine

Vaccination of chickens with an oil-emulsion vaccine containing a recombinant baculovirus that expressed the hemagglutinin-neuraminidase (HN) of Newcastle disease virus (NDV)-induced hemagglutination-inhibition (HI) and virus-neutralizing antibodies against NDV. HI antibody titers obtained in response to vaccination with the live recombinant virus were higher than those obtained when the recombinant was inactivated with beta-propiolactone, and the titers were lower than those obtained in response to the same HN concentrations in live or beta-propiolactone-inactivated NDV strain B1. The serological response to the recombinant baculovirus was differentiated from the response to NDV by an enzyme-linked immunosorbent assay in which purified NDV nucleoprotein was used as antigen. Chickens vaccinated with the live recombinant or with inactivated NDV resisted an oculonasal challenge with the neurotropic velogenic Texas GB strain of NDV, which was lethal in unvaccinated controls. It was concluded that the HN protein of NDV expressed as a subunit by a recombinant baculovirus was protective against Newcastle disease.