Use-Dilution Test and Newcastle Disease Virus
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The use-dilution test for evaluating the effectiveness of disinfectants against bacteria was modified to determine the effectiveness of disinfectants against a group of viruses. Modifications were kept to a minimum to retain the general principles of the test and thereby retain the test's familiarity among testing laboratory personnel. Modifications included the use of a standard allantoic fluid suspension of Newcastle disease virus instead of a standard bacterial culture. The only other modification was the inoculation of six embryonated chicken eggs (10 to 12 days old) with 0.1 ml of nutrient broth into which a carrier ring was transferred after a standard period in diluted disinfectant. The death or survival of 60 embryos, then, is the criterion by which a disinfectant can be judged effective at use-dilution. Experiments are described which establish the validity of the modified test procedure. The effectiveness of nine common disinfectants against Newcastle disease virus as judged by this test procedure is reported.Keywords:
Embryonated
Disinfectant
Newcastle Disease
Dilution
Serial dilution
Newcastle disease is one of the major economic threats to poultry population because of its high morbidity and mortality varying from 90-100%. It is caused by Avian Paramyxovirus-1 (APMV-1). This research work was carried out to identify Newcastle disease virus (NDV) by using reverse transcription-polymerase chain reaction (RT-PCR) assay and further isolate the virus in embryonated chicken eggs. A total of 127 cloacal swabs were collected from local chickens in live bird market and exotic chickens in commercial poultry farms in Zaria and environs, Nigeria between November, 2014 and January, 2015. Five commercial poultry farms and four live bird markets were purposively sampled. Molecular screening of NDV Matrix-gene (M-gene) was performed on all the samples using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). Newcastle disease positive samples were further inoculated into embryonated chicken eggs for isolation of Newcastle disease virus. Isolates were confirmed as Newcastle disease virus by haemaggulitination inhibition (HI) test. Newcastle disease virus Matrix-gene was detected in 16 (12.5%) out of 127 cloacal swabs; 13 (10.2%) from live bird markets and 3 (2.3%) from commercial poultry farms. However, only 10 Newcastle disease viruses were isolated in embryonated chicken eggs as confirmed by Haemagglutination inhibition (HI) test. Due to the higher detection rate recorded by reverse transcriptase-polymerase chain reaction (RT-PCR), it is therefore important that molecular technique be made easily accessible so that samples from each suspected outbreaks of NDV be screened so that rapid and confirmatory diagnosis can be achieved.Keywords: Embryonated chicken eggs, Haemagglutination inhibition test, Newcastle disease virus, RT-PCR
Embryonated
Newcastle Disease
Poultry farming
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The egg-adapted Newcastle disease virus (NDV) California strain used for this study was obtained in 1954 from the Bureau of Animal Industry, Department of Agriculture, Washington, D. C. This strain was isolated by the Bureau from infected chicken tissue from the corresponding state before the introduction of commercial live virus vaccines for Newcastle disease. The culture had been carried in 10-day embryonated chicken eggs through 28 serial passages. Embryo passage of the California strain was continued through the 29th passage. Allanto-amniotic fluid from several eggs dead of the strain was harvested and the strain of the virus was confirmed to be NDV by a neutralization test conducted in 10-day embryonated chicken eggs. The test was performed according to the procedure recommended by the Bureau of Animal Industry (U. S. Dept. Agri. Bull., 1946). Newcastle-immune chicken serum neutralized the virus in all cases while normal chicken serum had no effect. Reagan, …
Embryonated
Newcastle Disease
Strain (injury)
Virus strain
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Newcastle disease (ND) is one of the major viral diseases of poultry causing great economic losses to the poultry industry. This study was aimed to assess the epidemiological role of free living birds in the transmission of ND. A total of 63 cloacal swabs/droppings from various categories of free living birds (desi chicken, pigeon, turkeys, crows, sparrows, geese, parrots) were collected and inoculated into specific pathogen free embryonated chicken eggs for isolation of Newcastle disease virus (NDV). Four NDV isolates were obtained out of 63 cloacal swabs inoculated and the isolates were characterized as velogenic and lentogenic pathotypes based on mean death time (MDT) and intracerebral pathogenicity index (ICPI). It is concluded that free living birds may play an important role in the transmission of NDV to domestic chicken and enforce the biosecurity measures to minimize the effective contact between them.
Embryonated
Newcastle Disease
Biosecurity
Specific-pathogen-free
Cloaca
Poultry farming
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PREVIOUS studies have demonstrated the efficacy of immunizing chickens against Newcastle disease (ND) with B1 strain Newcastle disease virus (NDV) administered through the drinking water under laboratory and field conditions (Winterfield and Seadale, 1955Winterfield and Seadale, 1956Winterfield and Seadale, 1957a,b). In this investigation, F and LaSota NDV strains were used as vaccines in addition to the B1. The results are presented in this paper. MATERIALS AND METHODS B1 strain NDV (Hitchner and Johnson, 1948) vaccine was prepared from stock virus maintained in the authors’ laboratory. F strain NDV was obtained in a dry state from the Newcastle Disease Repository, University of Wisconsin, and the LaSota strain was isolated from a commercial vaccine. The F and LaSota strains were passed through three generations in embryonated chicken eggs before being used as vaccines. Each vaccine was in the form of amnioallantoic fluid (AAF) obtained from infected embryos. The same vaccine pool of each strain was used . . .
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Embryonated
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Strain (injury)
Attenuated vaccine
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RT-PCR for the detection of Newcastle disease virus (NDV) in allantoic fluids of SPF embryonated eggs as well as in tissues of SPF chickens infected experimentally is described. The method proved to be specific as all tested NDVs were detected and no cross reaction with other RNA viruses was observed. Sensitivity of the method was established at 10 5 ELD50/0.1 ml. To detect NDV in chicken tissues, SPF chickens were inoculated with 10 6 EID50 of 3 NDV reference strains: La Sota (lentogenic), Roakin (mesogenic) and Italy (velogenic pigeon variant) and 5 d p.i. various tissue samples were aseptically collected followed by RT-PCR and virus isolation on SPF embryos. The results showed high concordance: 93% (La Sota and Italy) to 100% (Roakin) between both methods.
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1. Data are presented to support the contention that the growth of Newcastle disease virus in the developing chick embryo occurs in cycles of 4 hr. The apparent 4-hr latent period in the production of virus following the inoculation of an infective dose does not correspond to the lag phase in the multiplication of bacterial cells, but rather indicates an intercellular development of virus particles that cannot be determined by any of the present technics for measuring viral activity. The maximum adsorption of virus onto susceptible cells occurs within 10 minutes after its introduction into the embryonated egg. 2. The existence of an unequal development of infective and hemagglutinating titers during the growth of Newcastle disease virus is described and an attempted explanation for this discrepancy is made on the hypothesis that the hemagglutination unit results from a maturation of the infectious particles and cannot therefore be detected early in its growth.
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Diagnosis of velogenic viscerotropic Ranikhet disease from six different flocks of desi chicken in and around Mumbai by gross and histopathological examination, isolation of virus and molecular methods.A total of 25 carcasses (varying between 2 and 6 carcasses from each flock) of six different flocks of adult desi chicken were subjected to necropsy examination for diagnosis of the disease during the span of a year (2014-2015). After thorough gross examination, the tissue samples were collected and processed for virus isolation and histopathological examination. The 20% tissue homogenate was inoculated into 9-day-old specific pathogen free (SPF) embryonated eggs. Mean death time (MDT) of embryos after inoculation and intracerebral pathogenicity index (ICPI) were used to judge velogenic nature of the virus. Newcastle disease virus (NDV) was isolated from six cases and confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) targeting the partial fusion protein gene of the viral genome.A total of 25 carcasses (varying between 2 and 6 carcasses from each flock) of six different flocks of desi chicken were presented for postmortem examination to Department of Veterinary Pathology, Bombay Veterinary College, Parel, Mumbai during 2014-2015. The gross and histopathological examination revealed lesions suggestive of viscerotropic velogenic form of the Newcastle disease (ND). The 20% tissue homogenate was inoculated into 9-day-old embryonated eggs from SPF chicken. NDV was isolated from six cases and confirmed by RT-PCR targeting the partial fusion protein gene. MDT of all the isolates was <60 h which indicated velogenic nature of the virus. ICPI of the isolates ranged between the 1.63 and 1.78. In four out of six outbreaks concurrent moderate to heavy infection of Ascardii galli in one flock and Railetina spp. in three flocks was also noted. In this study, viscerotropic velogenic form of ND was confirmed in all six outbreaks by gross and microscopic examination, virus isolation and RT-PCR.In this study, viscerotropic velogenic form of ND was confirmed in all six outbreaks by gross and microscopic examination, virus isolation and RT-PCR. Nowadays, vaccine strains Lasota, B1 and F strains are used widely in India to control the infection of NDV. However, virulent NDV strains are still isolated frequently in the birds under backyard and also in commercial venture which demonstrates that NDV remains an on-going threat to commercial as well as backyard poultry flocks.
Embryonated
Newcastle Disease
Flock
Gross examination
Histopathological examination
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Embryonated
Newcastle Disease
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In this study, we determined the suitability of lentogenic LaSota and naturally occurring avirulent I2 vaccine strains of Newcastle disease (ND) virus for efficacious in-ovo vaccination of broiler chickens. A total of 114 embyonated eggs divided into five groups (A, B, C, D and E) consisting of 25 eggs in each of groups A and B, 20 eggs in each of groups C and D and 24 eggs in group E were used in the study. Eighteen-day-old embryonated eggs in group A were vaccinated in-ovo with ND-I2 vaccine while the same age of embryos in group B were vaccinated with ND-LaSota. Thirteen-day-old embryonated eggs in groups C and D were vaccinated with ND-I2 and ND-LaSota respectively. Group E served as unvaccinated control. There was significant difference (p 2) used for the in-ovo vaccination were pathogenic for chick embryos, however, ND-I2 vaccine was better tolerated when administered to 18-day-old chick embryo.
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In ovo
Newcastle Disease
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Newcastle disease (ND) is one of the major viral diseases of poultry causing great economic losses to the poultry industry. A total of 63 cloacal swabs/droppings from various categories of free living birds (desi chicken, pigeon, turkeys, crows, sparrows, geese, parrots) were collected and inoculated into specific pathogen free embryonated chicken eggs for propagation of Newcastle disease virus (NDV). Four out of 63 cloacal swabs were positive for NDV by reverse transcriptase polymerase chain reaction for fusion protein cleavage site of NDV. It is concluded that free living birds may play an important role in the transmission of NDV to domestic chicken and strict farm biosecurity measures has to be adopted to minimize the effective contact between them.
Newcastle Disease
Embryonated
Biosecurity
Specific-pathogen-free
Poultry farming
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