Newcastle Disease Outbreaks Research Articles

Efficacy of Newcastle disease LaSota vaccine-induced hemagglutination inhibition antibodies against challenges with heterologous virulent strains of genotypes VII and IX

Newcastle disease (ND), caused by virulent Newcastle disease virus (NDV), still remains one of the most important avian diseases affecting the poultry industry worldwide, despite intensive vaccination programs have been implemented in many countries. All NDV isolates characterized to date are of one serotype and classified into classes I and II, with class II being further divided into twenty-one genotypes. Antigenic and genetic diversity is observed among the different genotypes. Current commercially available vaccines belonging to genotypes I and II are genetically divergent from strains that caused ND outbreaks worldwide in the last two decades. Reports of vaccination failures on their insufficient ability to inhibit infection or virus shedding have created renewed interest in developing vaccines homologous to virulent NDV circulating in the field. In this study, after vaccination with the most widely used LaSota vaccine (genotype II), chickens with different hemagglutination inhibition (HI) antibody levels were challenged with heterologous virulent NDV strains of genotypes VII and IX to evaluate how antibody levels relate to clinical protection and infection or virus shedding. Under the experimental condition, LaSota vaccine could fully protect birds from morbidity and mortality, but higher antibody levels were required to inhibit virus shedding. The number of birds shedding virus generally tended to decrease as the HI antibody titers increase in vaccinated birds. When the HI antibody titers reached ≥ 13 log2 and ≥ 10 log2, the virus shedding from JSC0804 strain (genotype VII) and F48E8 strain (genotype IX) could be completely inhibited, respectively, but it may be difficult to ensure that all individuals reach and maintain those levels in chicken flocks vaccinated according to routine procedure. Furthermore, the virus shedding in vaccinated birds was correlated with the amino acid similarity between the vaccine and challenge strains; more similarity, less virus shedding. The results obtained highlight that stringent biosecurity measures combined with vaccination are crucial for chicken farms to maintain a virulent NDV-free status.

Retrospective investigation of Newcastle disease reported in Türkiye between 2017-2019

This study was aimed to understand the spatial and seasonal epidemiology of Newcastle disease (ND) in Türkiye using the outbreak data between 2017-2019 and also to calculate the case-fatality rates of this disease. It was also aimed to produce the maps by using Geographical Information Systems (GIS). Data were obtained from the World Animal Health Information System (WAHIS) database of the World Organization for Animal Health (OIE). Total number of 220 outbreaks of ND were registered in 47 provinces of Türkiye between this years. Accordingly, 88,372 poultry birds transmitted the disease. The highest number of ND outbreaks, cases and deaths was reported in The Black Sea Region. According to the regions there was a statistically significant difference in the number of outbreaks (P<0.05), but there was no statistically significant difference in terms of cases and deaths (P>0.05). On the other hand the highest number of ND outbreaks, cases and deaths was reported in the spring season. As a result of the comparisons according to the seasons, there was a statistically significant difference in the number of deaths (P<0.05), but there was no statistically significant difference in terms of outbreaks and cases (P>0.05). The spatial and seasonal distributions identified in this study should be taken into account while attempting to control the disease. Also, it is thought that the creation of spatial maps based on ND outbreaks that are common in Türkiye will contribute to the determination of the areas where precautions should be taken against the disease.

Newcastle disease burden in Nepal and efficacy of Tablet I2 vaccine in commercial and backyard poultry production.

Poultry (Gallus domesticus) farming plays an important role as an income generating enterprise in a developing country like Nepal, contributing more than 4% to the national Gross Domestic Product (GDP). Newcastle Disease (ND) is a major poultry disease affecting both commercial and backyard poultry production worldwide. There were more than 90 reported ND outbreaks in Nepal in 2018 with over 74,986 birds being affected. ND is responsible for over 7% of total poultry mortality in the country. Recent outbreaks of ND in 2021 affected many farms throughout Nepal and caused massive loss in poultry production. ND is caused by a single-stranded ribonucleic acid (RNA) virus that presents very similar clinical symptoms as Influenza A (commonly known as bird flu) adding much complexity to clinical disease identification and intervention. We conducted a nationwide ND and Influenza A (IA) prevalence study, collecting samples from representative commercial and backyard poultry farms from across the major poultry production hubs of Nepal. We used both serological and molecular assessments to determine disease exposure history and identification of strains of ND Virus (NDV). Of the 40 commercial farms tested, both NDV (n = 28, 70%) and IAV (n = 11, 27.5%) antibodies were detected in majority of the samples. In the backyard farms (n = 36), sero-prevalence of NDV and IAV were 17.5% (n = 7) and 7.5% (n = 3) respectively. Genotype II NDV was present in most of the commercial farms, which was likely due to live vaccine usage. We detected never reported Genotype I NDV in two backyard farm samples. Our investigation into 2021 ND outbreak implicated Genotype VII.2 NDV strain as the causative pathogen. Additionally, we developed a Tablet formulation of the thermostable I2-NDV vaccine (Ranigoldunga™) and assessed its efficacy on various (mixed) breeds of chicken (Gallus domesticus). Ranigoldunga™ demonstrated an overall efficacy >85% with a stability of 30 days at room temperature (25°C). The intraocularly administered vaccine was highly effective in preventing ND, including Genotype VII.2 NDV strain.

Emergence of a novel genotype of class II New Castle Disease virus in North Eastern States of India

Outbreaks of New Castle Disease from three north eastern states of India were confirmed by clinico-pathological examination followed by reverse transcription-PCR detection of F gene of ND Virus (NDV). Irrespective of vaccination, the outbreaks resulted 90–100% mortality in the affected flocks. The analysis of fusion protein sequences from ten field isolates revealed them as the velogenic or highly virulent strain. Phylogenetic analyses based on the complete F gene nucleotide sequences of the isolates have characterized only one of the isolate (OK149201) in the genotype XIII.2.2. The rest of the nine isolates are depicted in a distinct monophyletic group with average nucleotide distances from the other 20 genotypes ranged from 10.90 − 20.70. The nine isolates were further divided into two sub branches with the bootstrap support value of 100% at the nodes that define the two subgroups with an average evolutionary nucleotide distance of 6.00between the isolates in the two subgroups. As per the recommendation put forth in recently updated unified phylogenetic classification system for NDV, our findings clearly indicates emergence of a novel genotype of class II NDV in the biodiversity hot spot region of NER, India. The isolates in the newly identified genotype is designated with next available Roman numerals XXII. Further, the two subgroups within the genotype are designated as XXII.1 and XXII.2.

Game Birds Can Act as Intermediaries of Virulent Genotype VII Avian Orthoavulavirus-1 between Wild Birds and Domestic Poultry.

Newcastle Disease (ND), caused by virulent forms of Avian orthoavulavirus serotype-1 (AOAV-1) is an economically important avian disease worldwide. The past two incursions of ND into the United Kingdom occurred in game bird populations during 2005 and 2006. The nature of the game bird semi-feral rearing system, which can bring these birds into close contact with both wild birds and commercial or backyard poultry, has been hypothesized to act as a bridge between these two environments. As such, the risk that AOAV-1-infected game birds may pose to the UK poultry industry was investigated. Pheasants, partridges and chickens were experimentally infected with the virulent strain APMV-1/Chicken/Bulgaria/112/13, a genotype VII.2 virus associated with ND outbreaks in Eastern Europe. The study demonstrated that both chickens and pheasants are susceptible to infection with APMV-1/Chicken/Bulgaria/112/13, which results in high mortality and onward transmission. Partridges by contrast are susceptible to infection, but mortality was reduced, as was onward transmission. However, the data indicated that both pheasants and partridges may serve as intermediate hosts of AOAV-1 and may bridge the wild bird-domestic poultry interface enabling transmission into an economically damaging environment where morbidity and mortality may be high.

Understanding the disease and economic impact of avirulent avian paramyxovirus type 1 (APMV-1) infection in Great Britain.

Newcastle disease (ND) is a notifiable disease affecting chickens and other avian species caused by virulent strains of Avian paramyxovirus type 1 (APMV-1). While outbreaks of ND can have devastating consequences, avirulent strains of APMV-1 generally cause subclinical infections or mild disease. However, viruses can cause different levels of disease in different species and virulence can evolve following cross-species transmission events. This report describes the detection of three cases of avirulent APMV-1 infection in Great Britain (GB). Case 1 emerged from the 'testing to exclude' scheme in chickens in Shropshire while cases 2 and 3 were made directly from notifiable avian disease investigations in chicken broilers in Herefordshire and on premises in Wiltshire containing ducks and mixed species, respectively). Class II/genotype I.1.1 APMV-1 from case 1 shared 99.94% identity to the Queensland V4 strain of APMV-1. Class II/genotype II APMV-1 was detected from case 2 while the class II/genotype I.2 virus from case 3 aligned closely with strains isolated from Anseriformes. Exclusion of ND through rapid detection of avirulent APMV-1 is important where clinical signs caused by avirulent or virulent APMV-1s could be ambiguous. Understanding the diversity of APMV-1s circulating in GB is critical to understanding disease threat from these adaptable viruses.

Molecular characterization of the Newcastle disease virus that caused an outbreak in backyard birds in Costa Rica in 2015

Costa Rica gained its Newcastle Disease Virus NDV-free status with vaccination according to OIE proceedings in 1996, and its declaration as a country free of the velogenic, viscerotropic form of this disease (G/SPS/GEN/119) presented to the World Trade Organization (WTO) in 1999. On April 24th, 2015, SENASA (National Animal Health Service) attended a velogenic Newcastle disease outbreak that affected backyard chickens in a small town (Bellavista, Guanacaste) in the northern part of the country, near the Nicaraguan border. Sixty-five backyard birds died from a total of 84 exposed animals. Blood samples, cloacal swabs, tracheal swabs, cecal tonsils, lung and trachea tissues were collected for diagnosis at the National Veterinary Services Laboratory (LANASEVE). These samples were screened for Avian Influenza (AIV) and NDV. All samples were negative for Avian Influenza in ELISA test and RT-PCR. Serum samples were positive for NDV antibody by hemagglutination inhibition test, and tissue and swab samples were positive for NDV by conventional RT-qPCR targeting a 310 bp fragment of the virus fusion protein gene. The amino acid sequence of the protease cleavage site within the amplicon matched the sequence of a virulent strain (112RRQKRF117). The nucleotide sequence had a 98.7% identity and an e value of 4e-153 with a genotype V velogenic sequence from Belize (KF767467) and Honduras (JN872194) collected in 2008 and 2007, respectively, according to BLASTN. A total of 3604 backyard birds were euthanized in town and its surroundings (1 km), including 3495 chickens, 66 turkeys, 6 geese, and 37 ducks. The case was considered resolved, and OIE was notified in November 2015 following OIE guidelines. In April 2017, Costa Rica recovered its disease-free status through executive decree No. 40301-MAG.

Identification of Newcastle Disease Virus sub-genotype VII 1.1 isolated from chickens in Sabah, Malaysia.

Newcastle disease (ND) is an extremely contagious and fatal viral disease causing huge economic losses to the poultry industry. Following recent ND outbreaks in Sabah in commercial poultry and backyard farms, it was speculated that this could be due to a new introduction of Newcastle Disease Virus (NDV) genotype/sub-genotype. Here we report the genetic characterization of NDVs isolated from Sabah during early 2021. All isolates were amplified and sequenced with primers specific to the viral fusion (F) gene using reverse transcription-polymerase chain reaction (RT-PCR). Nucleotide sequence analysis of the F gene showed that all isolates shared similar homology of 99.4% with NDV strain from Iran isolated in 2018. Amino acid sequences of the F protein cleavage site revealed the motif of 112RRQKRF117 indicating all isolates were of virulent strain. Phylogenetic analysis demonstrated that all isolates were clustered under sub-genotype VII 1.1 and clustered together with isolates from Iran (previously known as subgenotype VIIl). The present findings suggested that there is an emerging of a new sub-genotype into the poultry population in Sabah and this sub-genotype has never been reported before in Malaysia. Therefore, transboundary monitoring and continuous surveillance should be implemented for proper control and prevention of the disease. A further molecular epidemiological analysis of NDV is needed to well understand the circulatory patterns of virulent strains of NDV in the country to prevent future outbreaks.

Comparative pathogenesis of two genotype VI.2 avian paramyxovirus type-1 viruses (APMV-1) in pheasants, partridges and chickens

ABSTRACT Newcastle disease (ND) is caused by virulent forms of avian paramyxovirus-1 (APMV-1) and is an economically important disease of poultry world-wide. Pigeon paramyxovirus 1 (PPMV-1), a sub-group of APMV-1 is endemic in Columbiformes and can cause infections of poultry. An outbreak of ND in partridges in Scotland, UK, in 2006 (APMV-1/partridge/UK(Scotland)/7575/06) was identified as a class II, genotype VI.2.1.1.2.1, more commonly associated with PPMV-1. It has been hypothesized that game birds may be a route of transmission into commercial poultry settings due to the semi-feral rearing system, which potentially brings them into contact with both wild-birds and poultry species. Therefore, the pathogenesis and transmission of APMV-1/partridge/UK(Scotland)/7575/06 in game birds and chickens was investigated, and compared to a contemporary PPMV-1 isolate, PPMV-1/pigeon/UK/015874/15. Viral shedding and seroconversion profiles demonstrated that pheasants were susceptible to infection with APMV-1/partridge/UK(Scotland)/7575/06 with limited clinical signs observed although they were able to excrete and transmit virus. In contrast, partridges and pheasants showed limited infection with PPMV-1/pigeon/UK/015874/15, causing mild clinical disease. Chickens, however, were productively infected and were able to transmit virus in the absence of clinical signs. From the data, it can be deduced that whilst game birds may play a role in the transmission and epidemiology of genotype VI.2 APMV-1 viruses, the asymptomatic nature of circulation within these species precludes evaluation of natural infection by clinical surveillance. It therefore remains a possibility that genotype VI.2 APMV-1 infection in game birds has the potential for asymptomatic circulation and remains a potential threat to avian production systems. RESEARCH HIGHLIGHTS Demonstration of infection of game birds with Pigeon paramyxovirus-1 (PPMV-1). There are differing dynamics of infection between different game bird species. Differing dynamics of infection between different PPMV-1 isolates and genotypes in game birds and chickens.

Analysis of Amino Acid Changes in the Fusion Protein of Virulent Newcastle Disease Virus from Vaccinated Poultry in Nigerian Isolates.

The roles of fusion gene in the virulence of Newcastle disease virus are well established, but the extent of its variation among the XIV, XVII, and XVIII genotypes reported in Central Africa and West Africa has until recently been understudied. In this study, virulent Newcastle disease virus (vNDV) was isolated from dead chickens among vaccinated flocks between March and April 2020. Fusion (F) gene was sequenced and analysed for characterization and information about genetic changes. Many substitutions were observed along the region and some of their functions are yet to be determined. Results showed that all study isolates have virulent cleavage site sequence 112-RRRKR-116/F117 and clustered within genotype XIVb. Sequence analysis showed K78R mutation in the A2 antigenic epitope in all isolates and more along the F-gene which varied in some instances within the isolates. Mutation in this A2 antigenic epitope has been reported to induce escape mutation to monoclonal antibodies generated using the NDV LaSota strain. The range of percentage nucleotide and amino acid homology between the study isolates and commercially available vaccine strains is 81.14%-84.39% and 0.175-0.211, respectively. This report provides evidence of vNDV among vaccinated chicken flock and molecular information about circulating vNDV strains in Kano State, Nigeria, which is useful for the development of virus matched vaccines. Newcastle disease (ND) surveillance and molecular analysis of circulating strains in this region should be encouraged and reported. Furthermore, ND outbreaks or cases among vaccinated poultry presented to veterinary clinics should be reported to the state epidemiologist. Nucleotide sequences were assigned accession numbers OK491971-OK491977.

JMM Profile: Avian paramyxovirus type-1 and Newcastle disease: a highly infectious vaccine-preventable viral disease of poultry with low zoonotic potential.

Newcastle disease (ND) is a highly contagious disease of poultry caused by virulent avian paramyxovirus-1 (APMV-1) (previously termed avian avulavirus-1 and avian orthoavulavirus-1). APMV-1 is endemic in poultry in many developing countries, whilst outbreaks still occur in developed countries, affecting both commercial and backyard flocks. ND outbreaks can have substantial economic consequences due to high mortality rates and the imposition of trade restrictions. APMV-1 nucleic acid can be detected from swabs or tissues of suspected cases by PCR. Evidence of infection or vaccination may be demonstrated by the presence of specific antibodies against HN in serum samples. No anti-viral treatments exist, but vaccines are available, although there are currently concerns over their efficacy.

Screening of Healthy Feral Pigeons (Columba livia domestica) in the City of Zurich Reveals Continuous Circulation of Pigeon Paramyxovirus-1 and a Serious Threat of Transmission to Domestic Poultry.

Pigeon paramyxovirus-1 (PPMV-1) is predominantly isolated from pigeons or doves and forms a separate group of viral strains within Avian Orthoavulavirus-1, the causative agent of Newcastle disease in poultry. Since the introduction of PPMV-1 into Europe in 1981, these strains have rapidly spread all over Europe, and are nowadays considered to be enzootic in feral and hobby pigeons (Columba livia domestica). Infections with PPMV-1 can range from asymptomatic to fatal. To assess whether PPMV-1 continuously circulates in healthy feral pigeons, 396 tissue samples of pigeons from the city of Zurich were tested by reverse transcriptase real-time PCR over the period of one year. PPMV-1-RNA was detected in 41 feral pigeons (10.35%), determined as the dominant European genotype VI.2.1.1.2.2. In 38 of the 41 pigeons where organ samples tested positive, PPMV-1-RNA was also detected in either choana or cloaca swabs. There were no significant differences in positivity rates between seasons, age, and sex. The current study shows that feral pigeons without clinical signs of disease can harbour and most likely excrete PPMV-1. Spill-over into free-range holdings of chickens are therefore possible, as observed in a recent outbreak of Newcastle disease in laying hens due to PPMV-1 genotype VI.2.1.1.2.2. in the canton of Zurich in January 2022.

Outreach Efforts to Prevent Newcastle Disease Outbreaks in Southern California.

In May 2018, an outbreak of virulent Newcastle disease (vND) affected backyard and commercial premises in Southern California. The re-occurrence of these outbreaks since the 1970s suggests that some poultry communities may not have reliable and stable resources available regarding biosecurity and disease prevention. Therefore, staff at the University of California, Davis (UCD) School of Veterinary Medicine (SVM), and the California Department of Food and Agriculture (CDFA) began organizing educational events and learning more about the gamefowl breeder community through a needs assessment exercise, during which local feedstores and neighborhoods were also visited. Focus groups were organized with breeders in various cities within the regional quarantine area, established by the CDFA during the vND outbreak. The focus groups were aimed at creating open communication networks with gamefowl breeders in the affected area, as well as to learn about their current sources of information, learning preferences, and current management practices. With the input from gamefowl breeders, as well as funding and input from the CDFA and the United States Department of Agriculture (USDA), a quality assurance program called the “Gamefowl Wellness Program” was established. Educational content was created and published through the UCD Gamefowl Wellness Program poultry health website. Additionally, with the help of the CDFA, the USDA, and pharmaceutical companies, Newcastle disease vaccines and training for their application were provided to feedstores with gamefowl breeder clientele. Nurturing trust with these poultry communities allowed us to receive the information needed to develop effective outreach strategies that could better serve them. Responding to community concerns might be a way to garner the trust necessary to prevent or at least promptly detect foreign animal disease outbreaks.

A study of risk factors associated with Newcastle disease and molecular characterization of genotype XIII Newcastle disease virus in backyard and commercial poultry in Assam, India

The continuous emergence of Newcastle disease virus (NDV) poses a persistent threat to the poultry industry. Recent increasing outbreaks of NDV in the North East region of India have highlighted the need to closely monitor and analyze the potential risk factors for Newcastle disease (ND) outbreaks. In the present study, an attempt was made to genotype the circulating Newcastle disease virus (NDV) in the backyard and commercial poultry flocks in Assam, India. Sera samples from unvaccinated backyard poultry flocks and tissue samples of ND suspected cases were collected and tested for the presence of NDV antibodies using the Haemagglutination inhibition (HI) test. A total of seven NDV isolates were analyzed from different districts of Assam, India, both genotypically and pathotypically. All isolates were characterized as virulent, carrying 112RRKQRF117 amino acid residues at the cleavage site. As determined by phylogenetic analysis, the isolates clustered with members of genotype XIII of class II NDV. Further analysis of risk factors of ND occurrence was conducted through a questionnaire survey. All the results indicated an occurrence of genotype XIII of NDV in the farms with inadequate biosecurity and farming practices.

Vaccination indices and concomitant serological status of Newcastle disease in chickens in Aba and Umuahia of Abia State

The vaccination indices of Newcastle disease (ND) in chickens in Aba and Umuahia towns of Abia state were studied alongside their corresponding antibody status. A total of 296 sera samples were collected from 74 chicken farms. A Haemagglution inhibition (HI) test was conducted to determine the ND virus serum antibody levels. Open and closed-ended questionnaires were administered to staff on the farms selected randomly in the study areas. Information on vaccine types, origin, administering personnel, revaccination interval, and records of ND outbreaks was collected. Whereas 68.9% of the farm carried out vaccination on their own, 27.0% and 4.1% of the vaccination were done by veterinarians and animal health scientists Farms in Aba and Umuahia had average geometric mean titres (GMT) of 166.32 and 100.33, respectively. Approximately 87% of the farms had protective immunity (GMT >8 or log23) against ND. Chickens aged 1-3, 4-8, 9-16, 17 weeks and above had average GMTs of 64.00, 76.99, 283.7 and 197, respectively. Post-vaccination antibody titres were 128.92, 110.63, 52.07 and 43.65, after 1 week, 2-3 weeks, 4 weeks, and above 4 weeks, respectively. Indigenous ND vaccines had an average GMT of 182.55, while foreign ND vaccines had a GMT of 120.82. The Result showed that 77% of farmers used foreign vaccines whereas 23% used indigenous vaccines. On vaccination interval, 40.5% revaccinated for ND every three weeks, 21.6% monthly, 8.1% bi-monthly and 29.7% revaccinated when necessary. About 54% of farmers reported previous ND occurrence. This study identified high seroprevalence of ND antibodies in the flock studied and indicated a high level of awareness and adherence to NDV vaccination among the farmers in the study area. The local vaccines elicited better immunogenic responses than their foreign counterparts. We, therefore, recommend that usage of the local vaccines be adopted, and revaccination is done before a month interval.

Emergence of XIII.2.2 genotype of Avian Avulavirus-1 with unique FPCS site in India

In the present study, two isolates namely D162 and D165 obtained from Newcastle disease outbreaks inbackyard poultry in Tamil Nadu were subjected to phylogenetic analysis using complete Fusion (F) gene sequenceto understand the circulation and evolution of Avian Avulavius-1 (AAv1). Further, these isolates were pathotypedbased on mean death time and intra-cerebral pathogenicity index. Pathotyping and genotyping of D162 and D165showed that these isolates belonged to velogenic pathotype and XIII.2.2 sub-genotype. In addition to this, the Fusion Protein Cleavage Site (FPCS) of F gene was analysed for amino acid sequence pattern. The amino acid sequence of FPCS of both D162 and D165 was 112RRRKRF117, which is unique among XIII.2.2 genotypes of AAv1, whereas it is common among Pigeon Paramyxovirus. It was hypothesized that cross-species infection by AAv1 has led to emergence of isolates with increased virulence.

Isolation and Molecular Detection of Newcastle Disease Virus from Field Outbreaks in Chickens in Central Ethiopia.

BackgroundNewcastle disease is a major viral disease of poultry. The virus is a major problem for chickens in Ethiopia and there is a scarcity of updated information on the virological and molecular status of confirmation of Newcastle disease outbreak cases in the country.MethodsNewcastle disease outbreaks were investigated from February 2021 to October 2021 in central Ethiopia to isolate and detect the virus by cell culture and reverse transcriptase PCR. A total of 44 pooled tissue specimens were sampled from sick and recently dead chickens showing typical clinical signs of Newcastle disease. Virus isolation were performed using DF-1 cells and detection of the virus was done by real-time PCR.ResultsOut of 44 collected tissue samples, 38.63% (17/44) were positive on DF-1 cells. The result shows 17 of the clinically sick and dead chickens were positive for the virus by reverse transcriptase polymerase chain reaction. Based on the sample type, 54.54% (6/11) of the brain samples, 36.36% (4/11) of the intestines, 54.54% (6/11) of lung and trachea, 9% (1/11) of pooled liver, kidney, heart, and spleen samples were positive. Viruses were isolated in the proportions 37.5% (6/16), 25% (2/8), 50% (2/4), 25% (1/4), 50% (2/4) and 50% (4/8) from Sebeta, Bishoftu, Sululta, Nifas Silk, Kolfe and Yeka, respectively.ConclusionThis study showed that Newcastle disease is a major viral disease causing death of chickens in the study area. Therefore, any control approach should focus on the appropriate characterization of the virus strain causing the outbreak in the study area.

Molecular techniques are better choice as compared to conventional methods for pathotypic characterization of Newcastle disease virus in poultry birds

Newcastle disease (ND) has importance in the poultry industry across the world. Despite vaccination, outbreaks of ND have been reported from various parts of Pakistan. The purpose of current study was to characterize the Newcastle disease virus (NDV) through different methods i.e., molecular techniques and conventional methods and evaluate their efficiency. To isolate pathogenic NDV, samples were taken from the commercial poultry farms after an outbreak, while for the non-pathogenic NDV isolation commercially available Lasota vaccine was used. In the first step NDV was tested by F gene sequence through RT-PCR. The predicted sequence of the amino acid in fusion protein indicates the following motif 112R-R-Q-R-R-F117 in pathogenic and 112G-R-Q-G-R-L117 in non-pathogenic NDV strains. While in the second phase, the pathotypic characterization of NDV was done through conventional methods like Intracerebral pathogenicity index (ICPI) and Mean death time (MDT). The results from conventional methods were also in agreement with standard values. The findings of current study suggest that for the assessment of genetic nature and the rapid diagnosis of NDV; molecular techniques are better choice than conventional methods.

Detection of Newcastle disease virus and assessment of associated relative risk in backyard and commercial poultry in Kerala, India.

BackgroundNewcastle disease (ND) is an economically important viral disease affecting the poultry industry. In Kerala, a state in South India, incidences of ND in commercial and backyard poultry have been reported. But a systematic statewide study on the prevalence of the disease has not been carried out.ObjectivesA cross‐sectional survey was performed to detect the presence of Newcastle disease virus (NDV) in suspect cases and among apparently healthy commercial flocks and backyard poultry, in the state and to identify risk factors for NDV infection.MethodsReal‐time reverse transcription‐PCR (RT‐PCR) was used to detect the M gene of NDV in choanal swabs and tissue samples collected from live and dead birds, respectively and the results were statistically analysed.ResultsThe predominant clinical signs of the examined birds included mild respiratory signs, huddling together and greenish diarrhoea. Nervous signs in the form of torticollis were noticed in birds in some of the affected flocks. On necropsy, many birds had haemorrhages in the proventriculus and caecal tonsils which were suggestive of ND. Of the 2079 samples tested, 167 (8.0%) were positive for the NDV M‐gene by RT‐PCR. Among 893 samples collected from diseased flocks, 129 (14.5%), were positive for M gene with pairwise relative risk (RR) of 15.6 as compared to apparently healthy flocks where 6 out of 650 (0.9%) samples were positive. All positive samples were from poultry; none of the ducks, pigeons, turkey and wild birds were positive. Commercial broilers were at higher risk of infection than commercial layers (RR: 4.5) and backyard poultry (RR: 4.9). Similarly, birds reared under intensive housing conditions were at a higher risk of being infected as compared to those reared under semi‐intensive (RR: 6.7) or backyard housing (RR: 2.1). Multivariable analysis indicated that significantly higher risk of infection exists during migratory season and during ND outbreaks occurring nearby. Further, lower risk was observed with flock vaccination and backyard or semi‐intensive housing when compared to intensive housing. When the M gene positive samples were tested by RT‐PCR to determine whether the detected NDV were mesogenic/velogenic, 7 (4.2%) were positive.ConclusionsIn Kerala, NDV is endemic in poultry with birds reared commercially under intensive rearing systems being affected the most. The outcome of this study also provides a link between epidemiologic knowledge and the development of successful disease control measures. Statistical analysis suggests that wild bird migration season and presence of migratory birds influences the prevalence of the virus in the State. Further studies are needed to genotype and sub‐genotype the detected viruses and to generate baseline data on the prevalence of NDV strains, design better detection strategies, and determine patterns of NDV transmission across domestic poultry and wild bird populations in Kerala.

Circulation, genomic characteristics, and evolutionary dynamics of class I Newcastle disease virus in China

ABSTRACT Newcastle disease caused by Newcastle disease virus (NDV) is one of the most serious threats to chickens and has two clinical forms, typical and atypical, caused by velogenic and lentogenic strains, respectively. To control the epidemic, many vaccines against velogenic class II NDVs have been introduced worldwide, but this has led to accelerated mutation of class II viruses under immune pressure and, on the other hand, to non-vaccine targeting class I NDVs becoming the dominant population in poultry. In this context, this study provided the first large-scale genomic epidemiological and quasispecies dynamic analysis of class I NDVs in China, and found that class I viruses that first appeared in East and South China have spread to central China and become the dominant class with an average evolutionary rate of 1.797 × 10−3. In addition, single nucleotide polymorphism and intra-host single nucleotide variation analyses show that HN and P genes have high mutation rates and may act as front-runners for NDV to expand their host range and enhance their virulence. This study also found that the class I NDV population has accumulated a number of mutations under positive selection and that six isolates with shortened C-terminal extensions of the HN protein are evolving toward increased virulence. These results not only enrich the research resources but also help us to better understand the dynamic evolution and mutational trends of NDV at the genomic level, which is crucial for monitoring, early warning, and controlling the outbreak of Newcastle disease.