Studies of infectious laryngotracheitis vaccines: immunity in broilers

Broiler chickens were vaccinated at 18 days of age against infectious laryngotracheitis (ILT) using chicken-embryo-origin (CEO) and tissue-culture-origin (TCO) vaccines, each vaccine given either by drinking water, spray, or eyedrop. Controls were not vaccinated. The broilers were challenged 3 weeks later with virulent ILT virus (USDA challenge strain). Serum samples taken before challenge were analyzed by a virus neutralization (VN) test to determine titers due to vaccination. Both vaccines, regardless of route of administration, produced low VN titers, geometric mean titer (GMT) being less than 4.0 in all vaccinated groups. When administered by the same route, the CEO vaccine produced higher titers than the TCO vaccine. Titers following drinking-water or eyedrop administration of vaccines were higher than titers following spray vaccination. There was an inverse relationship between pre-challenge VN titers of groups of birds and the percentage of birds in the groups dying from ILT virus challenge. The drinking-water route of vaccination provided the most protection, while the spray provided the least.     

Comparison of a tissue-culture virus-neutralization test and the enzyme-linked immunosorbent assay for measurement of antibodies t infectious bronchitis

Two serological tests, the virus-neutralization (VN) test in tissue culture using a tissue-cell-adapted virus and the enzyme-linked immunosorbent assay (ELISA), were compared to detect antibodies against Massachusetts 41 and Connecticut 46 strains of Infectious Bronchitis Virus (IBV). The VN test was conducted in wells of microplates by the usual procedure. The two strains of IBV were adapted after 20 serial passages to induce CPE in 24 hours in chickens embryos kidney cells (CEKC). The ELISA test was carried out using partially virus following ultracentrifugation of each stain of IBV as antigen. The ELISA test detected higher geometric mean antibody titers (GMT) against both strains of IBV than did the VN test. One hundred four serum samples taken at 1, 3, 5, 9, 22, 24, and 26 weeks of age from a flock of chickens vaccinated with the Mass strain three times and the Conn strain of IBV two times during the growing period showed higher antibody titer responses to the Conn 46 than to the Mass 41 strain. Maternal antibodies in chicks one week of age were readily detected by the ELISA test, whereas low or insignificant titers were found by the VN test. Sera of vaccinated chickens collected following challenge with Mass 41 or Conn 46 strain of IBV showed that the ELISA was more sensitive and showed higher titers than did the VN test. Although the VN test showed no rise in GMT in the same sera tested with the heterologous virus, the ELISA showed a slight increase or cross-reaction. The serum samples from the unchallenged control group showed no change in GMT with either test or IBV strain.     

Studies on the live-virus vaccine against infectious laryngotracheitis of chickens. II. Evaluation of the tissue-culture-modified strain C7 in laboratory and field trials

Laboratory and field tests were conducted to evaluate the immunogenicity of a live-virus vaccine against infectious laryngotracheitis (ILT) of chickens that was prepared using tissue-culture-modified strain C7. Eighty-three percent or more of chickens vaccinated by the ocular (OC) or intranasal (IN) route with 10(4.75) TCID50 of the attenuated strain C7 at 50 days of age were protected against challenge with a virulent strain of ILT virus without showing any clinical signs for 4 weeks post-vaccination (PV). When vaccine was administered by aerosol, however, only 65% of vaccinated chickens were protected against challenge. Fifty-seven percent of chickens vaccinated at 70 days of age maintained immunity for 6 months PV. Immune response in younger chickens was inferior to that in older ones. In the field trials, clinical observation revealed no adverse reactions caused by the vaccination, and 60% or more of broilers and 80% or more of layers vaccinated by the OC route were protected against challenge at 4 weeks PV. These results confirmed the safety and efficacy of the vaccine.     

Safety and efficacy of the cell-associated vaccine prepared by an attenuated infectious laryngotracheitis virus.

The safety and efficacy of the cell-associated (C-A) vaccine prepared by chicken embryo fibroblast (CEF) cells infected with the tissue-culture-modified strain of infectious laryngotracheitis (ILT) virus were studied in chickens. Over seventy percent of chickens inoculated with the C-A vaccine by the subcutaneous (S.C.) or intramuscular (I.M.) route at 1 day of age was protected against challenge with a virulent strain of ILT virus without any clinical signs. Chickens vaccinated with the C-A vaccine at 1 day of age acquired immunity within 6 days after vaccination, and the protection rate maintained more than 60% until 10 weeks post-vaccination. The C-A vaccine was invariably effective for chickens at various age. There was no evidence that the development of immunity was hindered by further vaccination with Newcastle disease and infectious bronchitis combined live vaccine. In addition, the C-A vaccine was safe when chickens were inoculated with 10 doses. In the field trials of the C-A vaccine, no adverse reaction was observed, and over 65% of vaccinated chickens was protected against the challenge of the virulent ILT virus at 8 weeks after vaccination.     

Relationship among transmissible gastroenteritis virus antibody titers in serum, colostrum, and milk from vaccinated sows, and protection in their suckling pigs

We studied the antibody responses to transmissible gastroenteritis (TGE) in serum, colostrum, and milk from sows vaccinated with 2 attenuated (1 IM and 1 oral-IM) and 1 nonattenuated live vaccines and the relationship of these responses with the survivability of the sow's suckling pigs after challenge exposure with virulent TGE virus. Contrary to previous studies, the anti-TGE virus-neutralizing geometric mean titers (GMT) in the milk of sows vaccinated with attenuated vaccines at 3 and 5 days of lactation were similar to that found in the colostrum. Colostral and serum antibody titers were highest in sows given 2 injections of the IM attenuated vaccine. Half of the sows given the oral-IM attenuated vaccine did not seroconvert after 2 oral doses. Only sows vaccinated with the nonattenuated live vaccine had milk GMT that remained high for 21 days after farrowing. The linear relationship between colostral GMT and percentage of survivability of suckling pigs challenge exposed at 3 days of age was significant (P less than 0.05), although the relationship between serum GMT and percentage of survivability and the relationship between milk GMT and percentage of survivability were not significant (P greater than 0.10). The linear relationship between colostral (P less than 0.10) or pre-challenge exposure milk (P less than 0.05) GMT and percentage of survivability of suckling pigs challenge exposed at 5 days of age was significant.(ABSTRACT TRUNCATED AT 250 WORDS)     

抗禽流感或传染性喉气管炎两种重组鸡痘病毒疫苗联合免疫的相互影响

“表达H5N1禽流感病毒HA和NA基因的重组鸡痘病毒（rFPV—AI）疫苗”以及“表达传染性喉气管炎病毒gB基因的重组鸡痘病毒（rFPV—ILT）疫苗”均已在实现商业化生产。由于两种疫苗使用了相同的载体病毒,如何使用才能减少相互干扰而产生最好的免疫效果。本研究设计了三个试验组：1）免疫rFPV—AI后间隔4周接种rFPV—ILT;2）rFPV—AI和rFPV—ILT混合后接种;3）将rFPV—AI和rFPV—ILT分别在两个翅膀同时免疫。结果表明,试验鸡接种rFPV—AI后4周接种rFPV—ILT,对传染性喉气管炎病毒WG株攻击的保护率为60％（6／10）,低于rFPV—ILT单独免疫组的保护率（100％,10／10）;rFPV—AI和rFPV—ILT混合后免疫组对禽流感病毒强毒攻击的保护率为80％（8／10）,对传染性喉气管炎病毒强毒攻击的保护率为70％（7／10）,而rFPV—AI和rFPV—ILT分两点同时接种对两种病毒攻击均能产生完全保护。本试验结果建议将这两种相同载体的重组病毒疫苗分两点同时接种以避免相互之间的干扰。     

Protection induced by infectious laryngotracheitis virus vaccines alone and combined with Newcastle disease virus and/or infectious bronchitis virus vaccines

Two types of live attenuated vaccines have been used worldwide for the control of infectious laryngotracheitis virus (ILTV): 1) chicken embryo origin (CEO) vaccines; and 2) tissue culture origin vaccines (TCO). However, the disease persists in spite of extensive use of vaccination, particularly in areas of intense broiler production. Among the factors that may influence the efficiency of ILTV live attenuated vaccines is a possible interference of Newcastle Disease virus (NDV) and infectious bronchitis virus (IBV) vaccines with the protection induced by ILTV vaccines. The protection induced by CEO and TCO vaccines was evaluated when administered at 14 days of age alone or in combination with the B1 type strain of NDV (B1) and/or the Arkansas (ARK) and Massachusetts (MASS) serotypes of IBV vaccines. Two weeks after vaccination (28 days of age), the chickens were challenged with a virulent ILTV field strain (63140 isolate, group V genotype). Protection was evaluated at 5 and 7 days postchallenge by scoring clinical signs and quantifying the challenge virus load in the trachea using real-time PCR (qPCR). In addition, the viral load of the vaccine viruses (ILTV, NDV, and IBV) was quantified 3 and 5 days postvaccination also using qPCR. The results of this study indicate that the NDV (B1) and IBV (ARK) vaccines and a multivalent vaccine constituted by NDV (B1) and IBV (ARK and MASS) did not interfere with the protection induced by the CEO ILTV vaccine. However, the NDV (BI) and the multivalent (B1/MASS/ARK) vaccines interfered with the protection induced by the TCO vaccine (P < 0.05). Either in combination or by themselves, the NDV and IBV vaccines decreased the tracheal replication of the TCO vaccine and the protection induced by this vaccine, since the ILTV-vaccinated and -challenged chickens displayed significantly more severe clinical signs and ILTV load (P < 0.05) than chickens vaccinated with the TCO vaccine alone. Although NDV and IBV challenges were not performed, the antibody responses elicited by NDV and/or the IBV vaccinations were significantly reduced (P < 0.05) when applied in combination with the CEO vaccine.     

Cell-culture virus-neutralization test and enzyme-linked immunosorbent assay for evaluation of immunity in chickens against fowlpox

Two serological tests--the virus-neutralization (VN) test in chicken embryo fibroblasts (CEF) using a cell-culture-adapted virus, and the enzyme-linked immunosorbent assay (ELISA)--were used for evaluating the immune response in chickens against fowlpox virus. The VN test was conducted in 96-well tissue-culture plates using a fowlpox virus that was adapted to induce cytopathic effects (CPE) in CEF in 48 hr. The ELISA was carried out with an antigen prepared by precipitation of a cell-culture-propagated virus suspension with ammonium sulfate and concentration by centrifugation. A 0.1 M acetate buffer, pH 5, was used as the sensitizing solution for maximum specific binding of the antigen to the microplate plastic well. No antibodies were detected by the VN test in 228 serum samples taken from chickens at irregular intervals between 1 and 39 weeks of age, even though the birds were vaccinated against fowlpox at 13 weeks of age. However, in sera collected 4 weeks after a sample of laying hens was challenged with fowlpox virus, VN titers of 1/10 to 1/40 were detectable. On the other hand, significant antibody reactions were detected by the ELISA on sera from chickens during the growing period, following vaccination and challenge. Although no maternal antibodies were found at 1 week of age, a continuous increase in the mean ELISA titers to fowlpox was demonstrated during the entire experimental period. This study showed that the ELISA was considerably more sensitive and practical than the VN test.     

Comparison of a kinetic-based enzyme-linked immunosorbent assay (KELISA) and virus-neutralization test for infectious bursal disease virus. II. Decay of maternal antibody in progeny from white Leghorns receiving various vaccination regimens

A computer-assisted single-serum-dilution indirect kinetic-based enzyme-linked immunosorbent assay (KELISA) was used for quantitating the natural decay rate of infectious bursal disease virus (IBDV) maternal antibody in progeny obtained from white leghorn breeders. The KELISA results were compared with those of a standard virus-neutralization (VN) test. Pullets were subjected to two IBDV immunization regimens. Group 1 was vaccinated at weeks 0, 2, and 10 with two live vaccines in drinking water and at week 20 with an oil-emulsified (SC) IBDV vaccine, group 2 received only the first and last immunizations, and group 3 served as the unvaccinated control. Pullets were artificially inseminated at 28 weeks of age. Progeny chicks from each group were bled every other day for 47 days. Both KELISA and VN test detected linear relationship in the decay of maternal antibodies. The VN test detected no significant differences (P greater than 0.05) in the IBDV maternal antibody titers at day 1 or in the rate of decay between the progeny from groups 1 and 2. The VN maternal antibody titers decreased at a rate of 0.16 log2 titer per day. In contrast, KELISA revealed higher IBDV maternal antibody titers in day-old progeny from pullets vaccinated 4 times (log2 = 14.3). However, KELISA titers of progeny from this group decreased at a faster rate than titers of progeny from pullets vaccinated twice (0.20 vs. 0.13 log2 titer per day).     

基于新城疫病毒V蛋白鉴别鸡群感染与免疫的间接ELISA方法的建立

本研究以新城疫病毒(NDV)V蛋白羧基端结构域(Vc)的重组蛋白为包被抗原,建立了用于检测NDV V蛋白抗体的间接ELISA方法,并采用该方法检测了鸡群免疫或接毒后血清中的V蛋白抗体水平。结果显示:两组不同NDV灭活疫苗组在免疫后的3周内检测结果均为阴性;两组灭活疫苗免疫3周后再人工感染NDV强毒的鸡群,攻毒后第7、14和21 d,NDV阳性率分别为60%、80%、70%和50%、80%、70%;两组不同的NDV弱毒疫苗免疫组鸡群,仅在免疫后第21 d阳性率分别为20%和10%。以上结果表明,NDV疫苗免疫组与强毒感染组的V蛋白抗体阳性率存在明显差异,本方法可在群体水平上区分新城疫疫苗免疫与强毒感染鸡群,为NDV血清学诊断和流行病学调查提供了一种新的检测手段。     

Efficacy of live virus vaccines against infectious laryngotracheitis assessed by polymerase chain reaction-restriction fragment length polymorphism

The efficacy of four different commercial live vaccines (vaccines A, B, C, and D) against the infectious laryngotracheitis virus (ILTV) was assessed in specific-pathogen-free (SPF) chickens. SPF chickens were vaccinated intraocularly at 6 wk old with ILTV live vaccines and were challenged intratracheally with the N91B01 strain of virulent Korean ILTV 2 wk after vaccination. The immunity against ILTV live vaccines was assessed by the incidence of latent infection by the challenge virus in the chickens' tracheas and trigeminal ganglia, the reisolation rate of the challenge virus, and the clinical signs in the chickens challenged with the N91B01 strain of ILTV. The latent infection in chickens was assessed by nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Our data showed that the clinical signs and challenge virus isolation were negative in all chickens receiving four difference commercial ILTV live vaccines. The viral DNA of the vaccine strain, but not that of the challenge virus, was detected in chickens vaccinated with vaccine A by nested PCR-RFLP. The viral DNAs of both the vaccine and challenge strains were detected from chickens vaccinated with vaccines B, C, and D. This study showed that only vaccine A can protect chickens from latent infection with the field virulent ILTV. We speculate that the efficacy of infectious laryngotracheitis live vaccines to protect chickens from latent infection with virulent ILTVs can be assessed by nested PCR-RFLP analysis.     

禽流感油乳剂灭活疫苗的研究

将6种不同亚型的禽流感病毒（AIVH2N9、H3N8、H5N1、H5N2、H7N1、H9N2）分别接种鸡胚,收获尿囊液,经甲醛灭活,以矿物油为佐剂制成油乳剂灭活疫苗。疫苗接种4和8周龄SPF鸡,注苗后均无不良反应。每种亚型疫苗免疫后14天和21天攻毒保护率均达90％-100％。分别用H5N1和H9N2亚型灭活疫苗免疫8周龄SPF鸡、25和28周龄健康商品蛋鸡,免疫后7天产生免疫力,14天保护率达100％,21天后抗体达高峰,仔鸡接苗后最高血凝抑制（HI）几何平均滴度（GMT）为7.3-8.0log2,蛋鸡为8.0-10.5log2。免疫后180天,抗体效价不低于6.5log2。免疫后180天分别以AIV攻击,H5亚型疫苗组,用强毒攻击无一发病和死亡,对照鸡全部发病死亡;H9亚型疫苗组,对照鸡在攻毒后72小时停产,免疫鸡无一发病且产蛋正常,对同源攻毒的保护率达100％。     

Vaccinating chickens against avian influenza with fowlpox recombinants expressing the H7 haemagglutinin

OBJECTIVE: To evaluate the vaccine efficacy of a fowlpox virus recombinant expressing the H7 haemagglutinin of avian influenza virus in poultry. PROCEDURE: Specific-pathogen-free poultry were vaccinated with fowlpox recombinants expressing H7 or H1 haemagglutinins of influenza virus. Chickens were vaccinated at 2 or 7 days of age and challenged with virulent Australian avian influenza virus at 10 and 21 days later, respectively. Morbidity and mortality, body weight change and the development of immune responses to influenza haemagglutinin and nucleoprotein were recorded. RESULTS: Vaccination of poultry with fowlpox H7 avian influenza virus recombinants induced protective immune responses. All chickens vaccinated at 7 days of age and challenged 21 days later were protected from death. Few clinical signs of infection developed. In contrast, unvaccinated or chickens vaccinated with a non-recombinant fowlpox or a fowlpox expressing the H1 haemagglutinin of human influenza were highly susceptible to avian influenza. All those chickens died within 72 h of challenge. In younger chickens, vaccinated at 2 days of age and challenged 10 days later the protection was lower with 80% of chickens protected from death. Chickens surviving vaccination and challenge had high antibody responses to haemagglutinin and primary antibody responses to nucleoprotein suggesting that although vaccination protected substantially against disease it failed to completely prevent replication of the challenge avian influenza virus. CONCLUSION: Vaccination of chickens with fowlpox virus expressing the avian influenza H7 haemagglutinin provided good protection against experimental challenge with virulent avian influenza of H7 type. Although eradication will remain the method of first choice for control of avian influenza, in the circumstances of a continuing and widespread outbreak the availability of vaccines based upon fowlpox recombinants provides an additional method for disease control.     

Efficacy of oil-emulsion vaccines prepared with pigeon paramyxovirus-1, Ulster, and La Sota Newcastle disease viruses

Three strains of avian paramyxovirus-1 virus (PMV-1) were used to prepare four experimental monovalent oil-emulsion vaccines. A pigeon PMV-1 isolate (PPMV-1) and the Newcastle disease virus strains La Sota and Ulster were used to prepare four pools of beta-propiolactone-inactivated allantoic fluid for the vaccines. Groups of susceptible white rock chickens and racing homing pigeons were vaccinated subcutaneously with one of the vaccines, and their serologic responses were determined using the hemagglutination-inhibition (HI) test at frequent intervals up to 9 weeks postvaccination. Pigeons were challenged after 10 weeks with a virulent PPMV-1 isolate given intravenously, observed for signs of disease for 5 weeks, and then tested for secondary serologic HI responses. The HI responses were measured using the three strains of virus as HI test antigens. The titers were generally greater when the hemagglutination antigen used in the test was homologous with the antigen used to prepare the vaccine. All vaccines protected pigeons against morbidity and death but not against infection with the challenge virus. The shedding of PPMV-1 challenge virus from PPMV-1 vaccinates was greatly reduced 6 days after challenge.     

Embryo vaccination against Marek's disease with serotypes 1, 2 and 3 vaccines administered singly or in combination

Marek's disease virus (MDV) vaccines of serotypes 1 and 2 administered in 18-day-old embryonated eggs induced better protection against post-hatch challenge at 3 days with virulent MDV than vaccines given at hatch. Embryonal vaccination with a polyvalent vaccine containing equal quantities of serotypes 1 and 2 of MDV and serotype 3 virus (turkey herpesvirus, HVT) was also significantly more effective than post-hatch vaccination. These and earlier results indicate that protective efficacy of single or combined Marek's disease vaccine serotypes against post-hatch challenge at 3 days can be substantially improved if the vaccines are injected into 18-day embryos rather than at hatch. Injection of vaccines of serotypes 1 or 2 into embryonated eggs or hatched chicks did not cause detectable gross or microscopic lesions in chickens. Vaccine viruses of serotypes 1 and 2 could be isolated from spleen cells of chickens 1 week post-vaccination, and the titer of recoverable viruses was higher in chickens that received the vaccines at the 18th day of embryonation than in chickens vaccinated at hatch. Although embryo vaccination with HVT usually provided better protection than post-hatch vaccination against early post-hatch challenge with variant pathotypes of MDV, the protection was poor regardless of vaccination protocol. If challenge with variant pathotypes of MDV was delayed until embryonally or post-hatch HVT-vaccinated chickens were 21 days of age, protection of chickens by HVT was not enhanced. Thus, resistance induced by embryonal vaccination with HVT was qualitatively similar to that induced by post-hatch vaccination with this virus.     

AN ASSESSMENT OF THE AUSTRALIAN V4 STRAIN OF NEWCASTLE DISEASE VIRUS AS A VACCINE BY SPRAY, AEROSOL AND DRINKING WATER ADMINISTRATION

SUMMARY An Australian strain of Newcastle disease virus, was evaluated for use as a vaccine following its administration by drinking water, aerosol and spray to chickens at 1 and 21 days of age. Haemagglutination inhbition antibody was produced and persisted for 11 weeks. Aerosol vaccination induced higher levels of haemagglutination inhibition antibody than the other methods of vaccination. No respiratory disease was observed following vaccination. Chickens vaccinated by aerosol and spray were fully protected when challenged at 5, 7 and 11 weeks of age with virulent Newcastle disease virus. Mortality of 10 to 30 per cent was observed in chickens vaccinated by drinking water and intranasally following challenge.     

Effect of maternal immunity on the immune response to oral vaccination against rabies in young foxes.

OBJECTIVE: To determine effect of maternal antibodies on immune response to oral vaccination against rabies in young foxes. ANIMALS: 250 cubs from 48 vixens. PROCEDURE: Sera were obtained from cubs of 36 vaccinated (maternally vaccinated [MV+]) and 12 nonvaccinated (MV-) vixens between 23 and 71 days of age and tested for neutralizing antibodies. Seventy-one MV+ cubs and 33 MV-cubs were vaccinated orally with modified-live virus vaccine SAD B19. Geometric mean titer (GMT) was determined in these cubs approximately 21, 39, and 57 days after vaccination. In a subsequent experiment, 10 vaccinated MV+ cubs, 6 vaccinated MV- cubs, and 6 control cubs were challenge inoculated with virulent rabies virus approximately 100 days after vaccination. RESULTS: Serum GMT of nonvaccinated MV cubs (0.23 U/ml) was significantly greater than that of non-vaccinated MV- cubs (0.15 U/ml). The GMT of vaccinated MV+ cubs 21, 39, and 57 days after vaccination were 2.85, 2.11, and 0.79 U/ml, respectively, and were significantly less than those of vaccinated MV- cubs (12.19, 6.76, and 4.02 U/ml, respectively). All challenge-inoculated cubs with GMT < 0.5 U/ml succumbed to rabies. CONCLUSION AND CLINICAL RELEVANCE: Partially impaired immune response in cubs < 8 weeks old from vaccinated vixens causes insufficient protection against rabies. Inhibition of the immune response persists longer than the period during which maternal antibodies are detectable. Thus, oral vaccination campaigns for young foxes in areas where vaccination has been performed need to be reconsidered.     

Effects of infectious bursal disease on Marek's disease vaccination: suppression of antiviral immune response

Studies were made to determine whether infectious bursal disease virus (IBDV) infection would affect the response of chickens to turkey herpesvirus (HVT) vaccination in the development and level of HVT viremia and virus-neutralizing (VN) antibodies to HVT. The HVT viremia in the vaccinated chickens was not affected by IBDV, whether IBDV was inoculated simultaneously with HVT vaccination at one day of age or whether it was inoculated 3 weeks postvaccination with HVT. However, VN antibody response to HVT was significantly suppressed (P less than 0.001) when vaccinated chickens were exposed to IBDV either at the time of vaccination or at 3 weeks postvaccination. Such immunosuppression by IBDV of VN antibody response to HVT vaccination may result in a reduced antiviral immunity against Marek's disease virus.     

Immunisation of fancy chickens against Newcastle disease]

The German Regulation on Fowl plague which is in force since 1994 laid down that any chicken of all races and all hybrids must be vaccinated against Newcastle disease (ND) in a mode that an adequate immunity is achieved. Onset, duration, and resistance to challenge of immunity induced by vaccination is well documented in the scientific literature for hybrid chicken of the layer and meat types. These data prove also innocuity and efficacy of the registered vaccines. In contrast, only a few and incomplete data exist on the development of ND directed immunity in fancy chickens. The present study describes vaccinations of chickens of 14 different hobby breeds with live LaSota vaccine (conjunctival application of 10(6) embryo-infective dose50 per bird) and with an inactivated oil-emulsion vaccine (intramuscular application of 0.5 ml per bird) and subsequent intramuscular challenge infections using the highly virulent NDV strain Herts 33/66. Chickens of all 14 breeds tolerated the application of both vaccines. All fancy chickens reacted with the production of serum antibodies which were measured in the haemagglutination inhibition (HI) and virus neutralisation (VN) tests. According to the scientific literature, maximal antibody levels are reached in hybrid chickens between day 10 and 20 post vaccination. In contrast, in fancy chickens the antibody maxima are delayed to the seventh to eighth week post vaccination. All fancy chickens vaccinated either once with live LaSota virus or with live and inactivated vaccines resisted challenge with the highly virulent Herts 33/66 strain of NDV and did not develop any signs of disease. There are indications for gradual differences in susceptibility of different breeds of fancy chickens. The levels of non-specific neutralisation as measured in the virus neutralisation test differ between breed. Also, the viral content in tissues obtained from non-vaccinated but challenged birds differ markedly. It is concluded from the results of this study that fancy chickens can also successfully protected against Newcastle disease by using live and inactivated vaccines which are licensed for hybrid chickens. However, the optimal time for the detection of maximal antibody levels in fancy chickens is reached seven to eight weeks post vaccination.