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Umali D.V.,University of the Philippines at Los Banos | Umali D.V.,Yamaguchi University | Umali D.V.,Poultry Products Quality Control PPQC Co. | Ito H.,Tottori University | And 5 more authors.
Journal of Veterinary Medical Science | Year: 2014

Relatively little is known about the distribution of avian paramyxoviruses (APMVs) among wild birds in Japan. Surveillance of APMV in migratory waterfowl was conducted in the San-in region of western Japan during winters of 2006 to 2012. A total of 16 avian paramyxoviruses consisting of 3 lentogenic Newcastle disease viruses (NDVs), 12 APMV-4 and 1 APMV-8 were isolated from 1,967 wild-bird fecal samples. The results show that NDV and APMV-4 are relatively widely distributed among wild waterfowl that migrate to Japan from northern regions. Phylogenetic analysis revealed that there was no genetic relationship between the isolates from wild birds and domestic poultry in Japan. However, surveillance of APMVs in wild waterfowl needs to be conducted due to the pathogenic potential of these isolates in domestic poultry. © 2014 The Japanese Society of Veterinary Science. Source

Umali D.V.,University of the Philippines at Los Banos | Umali D.V.,Yamaguchi University | Umali D.V.,Poultry Products Quality Control PPQC Co. | Ito H.,Tottori University | And 7 more authors.
Virology Journal | Year: 2013

Background: Newcastle Disease (ND) is a highly contagious and economically devastating disease of poultry. At present, limited molecular epidemiological data are available regarding the causes of ND outbreaks in vaccinated commercial poultry farms. Knowing the genomic characteristics of Newcastle disease virus (NDV) infecting commercial poultry operations in spite of vaccination might give important insights on the infection dynamics of these viruses. In addition, molecular analyses at the subgenotype level and studies on the relationship of Japanese NDVs with other isolates from around the world are lacking. Therefore, in the present study, a molecular epidemiological investigation was conducted to characterize nine NDVs isolated from vaccinated commercial poultry flocks in five different Prefectures in non-epidemic areas of Japan between 1969 and 2002. Methods. Nucleotide sequencing and phylogenetic studies were performed to characterize the complete fusion (F)-protein gene, 3-prime end of the nucleoprotein (NP)-gene and 5-prime end of the RNA dependent RNA polymerase (L)-gene. Sequence data were compared with 180 NDV strains from GenBank representing different NDV genotypes and subgenotypes from different regions of the world at different time periods. Deduced amino acids were analyzed for homologies, recombination and mutation. Recombination events were estimated using Recombination Detection Program (RDP) version 3.44. Phylogenetic trees were constructed to determine evolutionary relationships among strains. Results: Mean death time (MDT: 48-56 hr), Intracerebral Pathogenicity Index (ICPI: 1.7-1.9) and deduced amino acid sequences of the F0 proteolytic cleavage site (§ssup§112§esup§RRQKR§ssup§116§esup§) revealed that all nine field isolates were velogenic. Phylogenetic analysis showed that these isolates could be classified into two genetic lineages and three sublineages namely genotypes VIa (lineage 4a), VId (lineage 4d) and VIId (lineage 5d). No recombination events were observed but a point mutation in one of the neutralizing epitope of the F-protein was identified in the field isolates from Japan. Conclusions: All field isolates from vaccinated commercial poultry in non-epidemic areas of Japan were part of much bigger outbreaks in provinces and regions and, in some cases, continents. In general, four ND panzootics occurred in Japan and that these outbreaks were mostly characterized by co-circulation of genetically distinct virus lineages due to involvements of infected wild birds. The point mutation identified in the field isolates from Japan may be due to escape from vaccine pressure. The identification of such mutation may be useful for future site-directed mutagenesis to understand the dynamics of NDV infection in vaccinated chickens. © 2013 Umali et al.; licensee BioMed Central Ltd. Source

Umali D.V.,University of the Philippines at Los Banos | Umali D.V.,Yamaguchi University | Umali D.V.,Poultry Products Quality Control PPQC Co. | Ito H.,Yamaguchi University | And 6 more authors.
Virus Genes | Year: 2014

The complete genome sequences of three strains of Newcastle disease virus (NDV) isolated from vaccinated commercial layer flocks in Japan in the span of three decades were characterized. All strains had genome lengths of 15,192 nucleotides consisting of six genes in the order of 3′-NP-P/V/W-M-F-HN-L- 5′. The general genomic characteristics of the Japanese field strains were consistent with previously characterized class II NDV, except for those belonging to early genotypes (genotype I-IV), which lack the six nucleotide insertion at nucleotide positions 1,648-1,653 of the nucleoprotein (NP) gene. Phylogenetic analysis showed that the Japanese strains could be classified into genotypes VIc and VIIe using the complete genome sequence and the complete coding sequence of the fusion (F) gene according to the unified NDV classification system. Characterization of functional domains and neutralizing epitopes of the F and hemagglutinin-neuraminidase (HN) proteins of Japanese field strains revealed a total of 31 amino acid substitutions, as compared to vaccine strains Ishii and B1, which were widely used in Japan. Although virus neutralization (VN) test showed that poor flock immunity due to vaccination failure or partial and non-uniform immunization maybe the major factors involved in the mechanism of breakthrough infection of the Japanese field strains, approximately two to threefold decrease in the VN titers of the field NDV strains possessing a point mutation (E347K or E347G) at the linear epitope of the HN protein was observed, as compared to vaccine strain B1 and field strain 2440/69, which lack the point mutation. This study may be a useful reference in characterizing future ND outbreaks in vaccinated chickens and as a genetic map for future investigations regarding vaccine designs, reverse genetics systems, and development of molecular diagnostic tools to prevent future ND outbreaks in vaccinated poultry flocks. © 2014 Springer Science+Business Media. Source

Umali D.V.,University of the Philippines at Los Banos | Umali D.V.,Yamaguchi University | Umali D.V.,Poultry Products Quality Control PPQC Co. | Ito H.,Tottori University | And 6 more authors.
Poultry Science | Year: 2015

In 2002, a commercial layer flock in Japan was initially diagnosed as being infected with infectious bronchitis (IB) based on clinical signs, virus isolation, and serological analysis but was later found to be atypically infected with velogenic Newcastle disease virus (NDV) following molecular diagnosis. The flock had slightly decreased egg production and an increased occurrence of soft-shelled eggs without significant mortality. IB-like viruses were isolated, which caused dwarfing and curling in 12-day-old chicken embryos. Ten years after this case, retrospective genetic analyses showed that apart from IB virus (IBV), the flock was also infected with NDV. Mean death time (MDT), intracerebral pathogenicity index (ICPI), and deduced amino acid sequence of the cleavage site of the fusion (F)-protein gene revealed that the NDV isolate was velogenic (112RRQKR116). These results indicate that poultry clinicians should look out for atypical velogenic ND, especially in vaccinated commercial chicken flocks, which may harbor hidden NDV infection. © 2015 Poultry Science Association Inc. Source

Umali D.V.,University of the Philippines at Los Banos | Umali D.V.,Poultry Products Quality Control PPQC Co. | Lapuz R.R.S.P.,University of the Philippines at Los Banos | Suzuki T.,Poultry Products Quality Control PPQC Co. | And 2 more authors.
Avian Diseases | Year: 2012

Rodents play a major role in the transmission and maintenance of Salmonella contamination cycles in poultry facilities. However, very limited field data are available regarding the transmission routes, infection cycle, and shedding patterns of Salmonella by naturally infected wild rodents from commercial layer farms. In this study, a total of 128 resident wild roof rats (Rattus rattus) were captured from a Salmonella-contaminated layer facility. All roof rats were divided into 51 laboratory cages, and weekly monitoring of Salmonella fecal shedding patterns was conducted for 53 wk. Seven roof rats from cages that were observed to frequently shed Salmonella were isolated in individual cages, and daily Salmonella monitoring was performed for 35 days. At the end of monitoring, each roof rat was euthanatized, and isolation of Salmonella from different organs was performed. Results of weekly monitoring of Salmonella showed that 21 of 51 cages (41.2) were positive for Salmonella Infantis, while two cages (3.92) were positive for Salmonella Enteritidis. Moreover, 11 cages were positive for Salmonella for at least two sampling weeks. Isolation of Salmonella from fecal droppings was mainly observed during the first 12 wk of captivity. The longest interval between two Salmonella-positive fecal dropping was 24 wk. In the daily Salmonella monitoring, only Salmonella Infantis was isolated from fecal droppings, in which the highest number of Salmonella Infantis organisms per fecal dropping was at 1 × 108 colony-forming units (cfu), while the lowest measured quantity was 1 × 103 cfu. It was noted that the frequency of Salmonella shedding in fecal droppings appeared to have a linear correlation (r 0.85) with the number of Salmonella organisms (cfu) per fecal pellet (P < 0.05). Moreover, pulsed-field gel electrophoresis analysis of Salmonella Infantis isolates revealed a single identical pulsed-field pattern. Salmonella Enteritidis isolates from fecal droppings and internal organs also generated a single identical pulsed-field pattern. Interestingly, Salmonella Infantis was not isolated from any of the organs examined, while Salmonella Enteritidis was isolated from the spleen and liver of one roof rat. These results may indicate that wild roof rats could persistently carry Salmonella and contaminate commercial poultry facilities through intermittent fecal shedding. Moreover, Salmonella Enteritidis in wild roof rats appears to be more of a systemic infection, in which isolation is most likely to occur in internal organs, whereas Salmonella Infantis is more likely an enteric type of infection, in which isolation is most likely to occur in the intestinal contents. It is very plausible that layer chickens could become infected with Salmonella through ingestion of Salmonella-positive fecal droppings or feeds contaminated with these fecal droppings from infected resident roof rats. This is likely one of the major reasons why layer houses can be persistently infected by Salmonella even if the facilities are thoroughly cleaned and disinfected and if replacement stocks are obtained from Salmonella-free breeders and rearing units. It is therefore a noteworthy suggestion that rodent control programs inside poultry premises comprise an essential and effective tool in the management and control of Salmonella contamination in layer flocks. © American Association of Avian Pathologists. Source

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