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.     

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.     

Antibody response and resistance of turkeys to Newcastle disease vaccine strain LaSota.

Studies were undertaken to determine the influence of repeated revaccination on the immune response in immuno-competent turkeys as measured by humoral antibody and resistance to challenge. Protection was better in turkeys given the LaSota spray vaccine at 4 weeks and 30 days later than in turkeys given one vaccination by spray or intramuscular route or exposed 4 times at 10-day intervals by the aerosol route. The anamnestic response, as measured by the HI tests to revaccination with the same immunogen, was not evident by the 3rd day postrevaccination but was observed on the 7th day. The interval between primary and secondary vaccination was found to be important to a true and optimal anamnestic response. Response was greater, however, in vaccinated turkeys exposed to VVND, a more virulent virus antigenically different from the vaccine strain. Exposure to LaSota vaccine by the intramuscular route gave a poorer HI response than LaSota given by aerosol.     

Characterisation of genotype VII Newcastle disease virus (NDV) isolated from NDV vaccinated chickens,and the efficacy of LaSota and recombinant genotype VII vaccines against challenge with velogenic NDV

A Newcastle disease virus (NDV) isolate designated IBS002 was isolated from a commercial broiler farm in Malaysia. The virus was characterised as a virulent strain based on the multiple basic amino acid motif of the fusion (F) cleavage site ¹¹²RRRKGF¹¹⁷ and length of the C-terminus extension of the hemagglutinin-neuraminidase (HN) gene. Furthermore, IBS002 was classified as a velogenic NDV with mean death time (MDT) of 51.2 h and intracerebral pathogenicity index (ICPI) of 1.76. A genetic distance analysis based on the full-length F and HN genes showed that both velogenic viruses used in this study, genotype VII NDV isolate IBS002 and genotype VIII NDV isolate AF2240-I, had high genetic variations with genotype II LaSota vaccine. In this study, the protection efficacy of the recombinant genotype VII NDV inactivated vaccine was also evaluated when added to an existing commercial vaccination program against challenge with velogenic NDV IBS002 and NDV AF2240-I in commercial broilers. The results indicated that both LaSota and recombinant genotype VII vaccines offered full protection against challenge with AF2240-I. However, the LaSota vaccine only conferred partial protection against IBS002. In addition, significantly reduced viral shedding was observed in the recombinant genotype VII-vaccinated chickens compared to LaSota-vaccinated chickens.     

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.     

Detection and differentiation of strains of Newcastle Disease Virus by complement fixation.

A complement-fixation test to detect Newcastle disease virus with antiserum produced in guinea pigs is described. Methodology is given for serum production and for standardization of the test. The test was used to differentiate 13 strains of Newcastle disease virus. Velogenic strains, including isolants form 1970-71 disease outbreaks in California, Florida, and Texas, were poor complement-fixing antigens, whereas lentogenic strains, including LaSota, Hitchner, and England F, were strong complement-fixing antigens. Mesogenic strains ranged from weak to strong in complement-fixing capabilities. This test can be used to differentiate velogenic field isolants from vaccine strains such as LaSota, Hitchner, and Roakin.     

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.     

Characteristics of paramyxoviruses isolated from pigeons in Czechoslovakia

Nine strains of paramyxovirus isolated from racing pigeons in southern Bohemia, Moravia and western Slovakia in 1983 were identified by the haemagglutination-inhibition test with antisera to seven types of paramyxovirus and three types of influenza A virus as PMV-1, Newcastle disease virus, in all cases. The haemagglutination activity and pathogenicity of the isolates for chicken embryos, chicken fibroblast cultures, and chickens of different age were determined. The mean death time of chicken embryos (MDT/MLD) was 52.8 to 95.4 h, the average being 75.7 h. The intracerebral pathogenicity index (ICPI8) was on an average 1.42 +/- 0.10 (P = 0.95). Experimental infection of chickens at the age of one, two, three and eight weeks did not cause any clinical disease but increased the level of haemagglutination-inhibiting (HI) serum antibodies up to 1 : 256 within three weeks. The course of heat inactivation of pigeon viruses at the temperature of 56 degrees C was practically identical with the inactivation of the velogenic viscerotropic strain California/1082/71. On the basis of the results, the pigeon isolates may be considered the Newcastle disease virus of velogenic viscerotropic type whose pathogenicity for chickens has been reduced to the level of mesogenic strains by long-time passaging in pigeons.     

Vaccination of ostriches (Struthio camelus, Linnaeus, 1758) against Newcastle disease: evidence for vaccine compatibility and seroconversion after vaccinations using the hemagglutination inhibition and virus neutralization tests

Newcastle disease (ND) remains to be the worldwide most important infectious disease of poultry. This epizootic is in Germany and many other countries a notifiable disease. Prophylactic vaccination is the major tool for the control of ND in poultry and other birds. Eighty-three ostriches (Struthio camelus) which were kept on farms in Germany were checked for the presence of NDV-specific antibodies. Some of these birds are said to be vaccinated against Newcastle disease. Only some of these ostriches contained antibodies which were measurable in haemagglutination inhibition and virus neutralisation tests. Twenty-three previously unvaccinated ostriches were vaccinated with commercially available vaccines. Both the LaSota live and inactivated oil emulsion vaccines were well tolerated following conjunctival or subcutaneous application, respectively. Neither local nor systemic side reactions were observed. After the vaccinations high antibody titres were detected in hemagglutination inhibition and virus neutralisation tests. A strong correlation between both established methods (r = 0.92; < 0.001) were noted.     

Immunization of day-old chicks having maternally derived antibodies against infectious bronchitis: degree of protection as monitored by ciliary activity after intratracheal challenge.

One-day-old chicks with maternally derived antibodies were vaccinated against infectious bronchitis (IB) with 3000 EID50 of the IB vaccine virus designated H120. The degree of protection induced by intranasal-eye drop (IE) vaccination was compared to that achieved by spray (S) vaccination. The protection afforded by vaccination was monitored by intratracheal challenge with IBV strain M-41 (clinical signs, ciliary activity in tracheal explants, virus isolation) and by serological tests (ovoneutralization, microneutralization in cell culture, haemagglutination inhibition (HI) test, ELISA). Intranasal-eye drop vaccination provided protection against intratracheal challenge. Immunity developed around 31 days of age. Spray vaccination failed to give protection against challenge by the same route. No difference was demonstrable in effectiveness between the two routes of vaccination by serological tests. No elevation of the antibody level occurred in either group. The level of maternally derived antibodies declined with age.     

Response of domestic geese to lentogenic and velogenic strains of Newcastle disease virus

A total of 54 domestic white meat-type geese were included in vaccination/challenge trials to evaluate susceptibility to disease and humoral immune responses using the haemagglutination inhibition (HI) and virus neutralization (VN) tests against Newcastle disease (ND). Two groups of twenty geese, five weeks of age, were conjunctivally vaccinated with either 100 x 10(6) or 2.5 x 10(6) EID50 (egg infectious dose 50 per cent) per bird of live La Sota virus, respectively, and 14 geese remained unvaccinated. At 15 weeks of age all vaccinated geese and seven unvaccinated geese were subcutaneously injected with 0.5 ml of inactivated oil emulsion ND vaccine, whereas seven geese remained as negative controls. At an age of 20 weeks, all 54 geese were challenged with 10(8.0) EID50 per bird of the viscerotropic velogenic NDV strain Herts 33/56. Live virus application as well as the oil emulsion vaccine did not induce discernible clinical signs and have no detrimental effect on body weight gains. At days 1, 3, 5, 8, 13, 16, 20, 23 and 27 after the application of lentogenic vaccine pharyngeal and cloacal swabs were taken, after challenge samples were taken at days 2, 5 and 8. Lentogenic as well as velogenic virus were never reisolated. Low and shortlived antibody responses post vaccination were equally well measured in HI and VN tests. Only two out of seven unvaccinated but challenged geese developed signs of ND whereas all vaccinated/challenged geese remained normal but developed high to moderate levels of HI and VN antibodies. Since domestic geese do not readily excrete NDV's in detectable amounts and since they do not contain detectable amounts of the challenge virus fourteen days post challenge in their tissues the assumption is promoted that geese do not play a major role in the epidemiology of Newcastle disease.     

Avian paramyxovirus serotype 1 isolates from the spinal cord of parrots display a very low virulence

The spinal cord of 32 psittacines suffering from proventricular dilatation disease (PDD) was investigated. In six cases, a virus was isolated which upon electron microscopic examination revealed morphological details typical of members of the Paramyxoviridae. All isolates were subsequently characterized as avian paramyxovirus serotype 1 (APMV-1) by type-specific polyclonal antisera. According to their reactivity with APMV-1 specific monoclonal antibodies, the six isolates shared epitopes within the haemagglutinin-neuraminidase spike protein, distinct from pigeon-type paramyxoviruses and the LaSota vaccine strain. This grouping was further corroborated by properties of the haemagglutinin: all isolates showed a very thermosensitive haemagglutination activity and were rapid eluters. Virulence of the APMV-1 isolates in 1-day-old specific pathogen free (spf) chicken was very low, with intracerebral pathogenicity indices between 0 and 0.1. In embryonated spf chicken eggs, psittacine isolates replicated to high titres (10(8.6)-10(10.7) EID50/ml). However, they exhibited a reduced lethality over an observation time of 7 days (10(6.1)-10(8.3) ELD50/ml). In a haemagglutination inhibition test with parrot sera from birds with no history of APMV-1 vaccination, sera reacted preferentially with two isolates compared with APMV-1 vaccine strains LaSota and B1. The other four isolates exhibited a differentiated reaction pattern with the parrot sera, indicating an antigenic inhomogeneity. This is the first report of isolating very low virulent APMV-1 from neuronal tissue of parrots and implications for a possible role in slow progressing disease will be discussed.     

Antibody response to strain combinations of Newcastle disease virus as measured by hemagglutination-inhibition

Differences in antibody response to three Newcastle disease virus (NDV) strains--B-1, LaSota, and Ulster--were investigated using the hemagglutination-inhibition (HI) micro-titer test in chickens hatched from ND-immune and unimmune flocks. When used singly as primary vaccines, the Ulster strain stimulated the lowest antibody response of the three in both immune and unimmune (susceptible) chickens. Subgroups of each of the primary-vaccinated groups were revaccinated with each of the three strains. Ulster-vaccinated chicks, revaccinated with Ulster, gave the poorest booster response. All other revaccination combinations gave a significant titer increase, though some were better than others. It is suggested that the Ulster strain as primary vaccine followed by booster does of B-1 or LaSota will induce a higher antibody response (i.e., immunity) in susceptible chicken populations with less risk of a post-vaccination reaction.     

Antigenic relationship among strains of ibaraki virus and epizootic haemorrhagic disease virus studied with monoclonal antibodies

Twelve monoclonal antibodies against ibaraki virus (IbV) were established and preliminarily characterised by indirect immunoperoxidase (IIP), haemagglutination inhibition (HI) and neutralisation (NT) tests. Five antibodies reacted in the IIP test with all IbV and epizootic haemorrhagic disease virus (EHDV) strains tested, and five antibodies reacted with IbV and Alberta strain (serotype 2) but not with New Jersey strain (serotype 1) of EHDV. Two of 12 antibodies showed both HI and NT activities. Viral proteins with molecular weights of about 24,000 daltons (24KD) and 78KD were determined by two monoclonal antibodies in Western blot analysis. One of two antibodies with the ability of both HI and NT recognised a viral protein with a molecular weight of about 78KD.     

Variation in immunogenicity of ruminant pestiviruses as determined by the neutralisation assay

Immunogenicity in naive three-month-old Friesian bull calves of nine ruminant pestiviruses, three each of type 1 bovine virus diarrhoea virus (BVDV), type 2 BVDV and border disease virus (BDV) was directly compared in reciprocal cross-neutralisation tests using sera obtained eight weeks after intranasal and intravenous inoculation with live virus. Cytopathic (CP) type 1 BVDV strain C86, non-cytopathic (NCP) type 2 BVDV strain 890 and NCP BDV strain V2536/2 were found to elicit significantly broad cross-neutralising antibodies against viruses in other species whereas other virus strains in all three species produced a much more pronounced homologous and/or species specific response. Results are clearly relevant in the selection of strains for vaccines against diseases caused by these successful, economically important ubiquitous viruses.     

Humoral antibody response and assessment of protection following primary vaccination of chicks with maternally derived antibody against avian infectious bronchitis virus

Commercially bred chicks with maternally derived antibody to avian infectious bronchitis virus (IBV) were hatched in isolated conditions and a number vaccinated within the first three weeks of life with live IBV strain H120. Humoral antibody responses were assayed by haemagglutination inhibition (HI) or neutralisation (SN) tests, and the degree of protection against challenge with the virulent Massachusetts M41 strain assessed on the basis of tracheal ciliary activity four days after challenge. Maternal antibody in unvaccinated chicks declined linearly with a mean half-life of five to six days based on both HI and SN tests; these chicks were protected against challenge until four weeks old. There was complete correlation between ciliary activity and histopathological findings, but little between protection and antibody titre. It was concluded that the optimum age for primary vaccination was about two weeks.     

一株鸭源新城疫病毒毒力基因分析及其对鸭的致病性研究

为了解中国鸭源新城疫病毒(Newcastle disease virus,NDV)的毒力特点,从2009年广东地区发病鸭群中分离和鉴定出1株新城疫病毒(简称NDV-104),对其生物学特性、致病性和融合蛋白(fusion,F)、血凝素神经氨酸酶(hemagglutinin-neuraminidase,HN)基因进行了研究。结果显示,分离株的MDT、ICPI和IVPI分别为56.4 h、1.95和1.64,结合F蛋白裂解位点(112~117位)的氨基酸序列分析,确定了分离株为新城疫强毒。致病性结果表明,分离病毒对雏鸭具有感染性和致病性。F基因遗传进化结果显示,分离株属于基因Ⅶd亚型。F、HN蛋白的氨基酸同源性结果表明,分离株与2000年以来国内外分离到的基因Ⅶ型NDV同源性较高,分别为96.6%~99.3%和97.0%~99.7%,而其与常用疫苗株B1、V4、Clone30、Mukteswar和LaSota的F、HN蛋白氨基酸序列的同源性较低,且与LaSota株和V4株的同源性最低,分别仅为88.1%和87.6%,说明分离株与经典新城疫病毒毒株存在一定的差异。     

蝙蝠作为流行性乙型脑炎病毒宿主的研究

1986、1988、1990和1997年的7－8月，在云南省共捕获653只蝙蝠，其中棕果蝠（Rousettus leschenaulti)593只，金管鼻蝠（Murina aurata)60只，采集血清及剖取脑组织作病毒分离和抗体检查。用细胞法和乳鼠法分离到6株病毒，带毒率为0．96％，其中从捕自景洪的棕果蝠脑组织中分离到3株，从河口的棕果蝠血清中分离到2株，从捕自耿马的金管鼻蝠脑组织中分离出1株。棕果蝠和金管鼻蝠的带毒率分别为0．84％和1．67％。这6株病毒能引起C6／36和BHK21细胞病变和导致乳小白鼠或3－4周龄小白鼠发病和死亡，并具有典型的嗜神经病毒感染症状，经血凝抑制，补体结合，免疫荧光和中和试验鉴定。新分离的6株病毒均为乙脑病毒（JE virus)。同期用血凝抑制试验对采获的425份棕果蝠血清作乙脑病毒抗体检查。阳性153－份，阳性率为36％，人有较高的感染率，。本次从棕果蝠和金管鼻蝠体内分离出乙脑病毒属具有重要流行病学意义。     

Generation of a genotype VII Newcastle disease virus vaccine candidate with high yield in embryonated chicken eggs

To generate a genotype VII Newcastle disease virus (NDV) vaccine with high yield in embryonated chicken eggs, we selected genotype VII NDV strain JS5/05, which possesses a high virus titer in embryos as the parental virus. Using reverse genetics, we generated a genetically tagged derivative (NDV/AI4) of JS5/05 by changing the amino acid sequence of the cleavage site of the F0 protein. Pathogenicity tests showed that NDV/AI4 was completely avirulent. NDV/AI4 was genetically stable and replicated efficiently during 10 consecutive passages in embryos. More importantly, serologic assays showed that oil-emulsion NDV/AI4 induced higher hemagglutination inhibition (HI) titers against the prevalent virus than oil-emulsion LaSota vaccine in chickens and geese. Moreover, NDV/AI4-induced HI titers rose faster than those elicited by LaSota in chickens. Both NDV/AI4 and LaSota provided protection against clinical disease and mortality after the challenge with the genotype VII NDV strain JS3/05. However, NDV/AI4 significantly reduced virus shedding from the vaccinated birds compared to LaSota. Taken together, these results suggest that NDV/AI4 can provide better protection than LaSota and is a promising vaccine candidate against genotype VII NDV.