“…Positive samples obtained in this study can further be subjected to pathotyping and phylogenetic studies which is an advantage that goes beyond the previous serological study. Even though, the distribution of the disease differs among the study areas in Eastern Shoa Zone using qRT-PCR analysis with statistically significance difference (p < 0.05), [12] reported an overall prevalence of 26.7% by using rRT-PCR for fussion (F) gene assay from Adama and Bishoftu which is in a close resemblance with our current findings (21.42%) when it was analyzed only for Adama and Bishofitu. In addition, using a rRT-PCR for M gene study conducted by [11] in three selected districts (Batu, Bishoftu and TikurWuha) in rift valley areas revealed an overall prevalence of 8.4% which is less than the current finding and this could be due to the small area coverage.…”
Section: Test Agreement Results Between Qrt-pcr and Rrt-pcr Detection supporting
confidence: 83%
“…The choice of matrix gene detection in NDV screening is due its highly conserved nature in the NDV genome where matrix gene assay is able to detect most of NDV isolates [10]. Despite the wide spread problem of ND in Ethiopia in general and Oromia in particular [7,11,12], there is limited information about the genetic profile of NDVs circulating in backyard poultry in Ethiopia. Therefore, the aim of this study was to detect the existence of Newcastle Disease virus by molecular tool in non-vaccinated village poultry in central rift valley of Oromia, Ethiopia.…”
Background Newcastle disease (ND) is a major infectious disease of poultry caused by a virulent strain of Avian Paramyxovirus – 1. It is a major threat to the poultry industry in many countries of the world including Ethiopia. Newcastle Disease Virus (NDV) is an enveloped, non-segmented, single-stranded negative-sense RNA virus with a helical morphology whose genome has six open reading frames (ORF) which encode for the following proteins: nucleoprotein (NP), phosphoprotein (P), matrix protein (M), fusion protein (F), hemagglutinin-neuraminidase (HN) and RNA-dependent RNA polymerase (L). The aim of this study was to detect matrix gene (M-gene), for NDV by molecular tools and identify its risk factors in non-vaccinated village chicken in Central Rift Valley of Oromia, Ethiopia.Methods A total of 84 pooled in five swab samples from 420 cloacal and tracheal chickens were sampled and RNA was extracted from the 84 pooled samples to carry out real-time quantitative polymerase chain reaction (qRT-PCR). A real-time reverse transcriptase polymerase chain reaction (rRT-PCR) along with the quantification was also done for 10 positive qRT-PCR samples that were having high concentration of viral load Ct. value.Results Out of the 84 pools of five swab samples tested for M-gene using qRT-PCR, 16.7% (14/84) samples were detected which included 13 positives and 1 negative for NDV. The prevalence of ND in males was found to be 16.10% and that in females was 14.67%. Although the overall ND prevalence was 15.48% (13/84), the highest score was recorded in Adama, 42.86% (6/14), and no positive case was detected in Bote and Bishoftu (p <0.05) while intermediate scores were obtained from Batu, Arsi-negelle and Shashemene.Conclusions In general, the present study provides important information on the epidemiology of NDV based on M-gene assay in Central Rift Valley of Oromia, Ethiopia, and highlights the importance of implementing surveillances and biosecurity practices in live poultry markets and village chickens.
“…Positive samples obtained in this study can further be subjected to pathotyping and phylogenetic studies which is an advantage that goes beyond the previous serological study. Even though, the distribution of the disease differs among the study areas in Eastern Shoa Zone using qRT-PCR analysis with statistically significance difference (p < 0.05), [12] reported an overall prevalence of 26.7% by using rRT-PCR for fussion (F) gene assay from Adama and Bishoftu which is in a close resemblance with our current findings (21.42%) when it was analyzed only for Adama and Bishofitu. In addition, using a rRT-PCR for M gene study conducted by [11] in three selected districts (Batu, Bishoftu and TikurWuha) in rift valley areas revealed an overall prevalence of 8.4% which is less than the current finding and this could be due to the small area coverage.…”
Section: Test Agreement Results Between Qrt-pcr and Rrt-pcr Detection supporting
confidence: 83%
“…The choice of matrix gene detection in NDV screening is due its highly conserved nature in the NDV genome where matrix gene assay is able to detect most of NDV isolates [10]. Despite the wide spread problem of ND in Ethiopia in general and Oromia in particular [7,11,12], there is limited information about the genetic profile of NDVs circulating in backyard poultry in Ethiopia. Therefore, the aim of this study was to detect the existence of Newcastle Disease virus by molecular tool in non-vaccinated village poultry in central rift valley of Oromia, Ethiopia.…”
Background Newcastle disease (ND) is a major infectious disease of poultry caused by a virulent strain of Avian Paramyxovirus – 1. It is a major threat to the poultry industry in many countries of the world including Ethiopia. Newcastle Disease Virus (NDV) is an enveloped, non-segmented, single-stranded negative-sense RNA virus with a helical morphology whose genome has six open reading frames (ORF) which encode for the following proteins: nucleoprotein (NP), phosphoprotein (P), matrix protein (M), fusion protein (F), hemagglutinin-neuraminidase (HN) and RNA-dependent RNA polymerase (L). The aim of this study was to detect matrix gene (M-gene), for NDV by molecular tools and identify its risk factors in non-vaccinated village chicken in Central Rift Valley of Oromia, Ethiopia.Methods A total of 84 pooled in five swab samples from 420 cloacal and tracheal chickens were sampled and RNA was extracted from the 84 pooled samples to carry out real-time quantitative polymerase chain reaction (qRT-PCR). A real-time reverse transcriptase polymerase chain reaction (rRT-PCR) along with the quantification was also done for 10 positive qRT-PCR samples that were having high concentration of viral load Ct. value.Results Out of the 84 pools of five swab samples tested for M-gene using qRT-PCR, 16.7% (14/84) samples were detected which included 13 positives and 1 negative for NDV. The prevalence of ND in males was found to be 16.10% and that in females was 14.67%. Although the overall ND prevalence was 15.48% (13/84), the highest score was recorded in Adama, 42.86% (6/14), and no positive case was detected in Bote and Bishoftu (p <0.05) while intermediate scores were obtained from Batu, Arsi-negelle and Shashemene.Conclusions In general, the present study provides important information on the epidemiology of NDV based on M-gene assay in Central Rift Valley of Oromia, Ethiopia, and highlights the importance of implementing surveillances and biosecurity practices in live poultry markets and village chickens.
“…Even though the distribution of the disease differs among the study areas in Eastern Shoa Zone using RT-qPCR on M gene assay with statistically significance difference ( P < 0.05), Miressa et al. (2016) reported an overall prevalence of 26.7% by using real-time reverse transcription polymerase chain reaction for F gene assay from Adama and Bishoftu.…”
Section: Resultsmentioning
confidence: 86%
“…The choice of M gene detection in APMV-1 screening is due to its highly conserved nature in the APMV-1 genome that enables to detect most of APMV-1 isolates ( Bello et al., 2018 ). Despite the wide spread problem of ND in Ethiopia in general and Oromia in particular ( Chaka et al., 2012 ; Terefe et al., 2015 ; Miressa et al., 2016 ), there is no information on M gene–based study about the status of APMV-1 circulating in backyard poultry in Central Rift Valley of Oromia, Ethiopia. Therefore, the aim of this study was to conduct molecular surveillance of ND virus based on M gene assay and identify potential risk factors for nonvaccinated village chicken in central rift valley of Oromia, Ethiopia.…”
Newcastle disease (
ND
) is a major infectious disease of poultry caused by a virulent strain of Avian Paramyxovirus type-1 (
APMV-1
). It is a major threat to the poultry industry in many countries of the world including Ethiopia. The aim of this study was to conduct molecular surveillance of ND Virus and identify potential risk factors for nonvaccinated village chicken in Central Rift Valley of Oromia, Ethiopia. A total of 84 pooled swab samples, each made from pools of 5 swabs for analysis, from cloacal and tracheal sites of chickens in the Central Rift Valley were collected, and RNA was extracted to carry out real-time quantitative polymerase chain reaction. Out of the 84 pooled swab samples tested for M-gene, 13 (15.48%) samples were found positive for APMV-1. The prevalence of ND in males was found to be 16.10% and that in females was 14.67%. Although the overall ND prevalence was 15.48% (13/84), the highest prevalence was recorded in Adama, 42.86% (6/14), and no positive case was observed in Bote and Bishoftu (
P
< 0.05), while intermediate prevalence was obtained from Batu, Arsi-negele, and Shashemene (
P
> 0.05). In general, the present study provides important information on the epidemiology of ND based on M-gene assay in Central Rift Valley of Oromia, Ethiopia, and highlights the importance of implementing molecular surveillances practice in live poultry markets and village chickens.
“…Similarly, our analysis revealed that NDV isolates of ON033824, ON033823, and ON033821 clustered with the genotype I, II, and III of NDV strain. Figure 2 Phylogenetic relationships between the Ethiopian NDV strain genotype (with accession number: ON033821, ON033822, ON033823, ON033824, and ON033825) and reference sequences with accession number: GU182327.1, AY175710.1, AY935490.1, GQ338309.1, JX129807.1, and KC851848.1, 24 the out group accession number: HM357606.2, 18 based on the partial F gene region nucleotide sequence by MEGA 10.2.4. …”
Background
Newcastle disease (ND) is a highly infectious poultry disease that causes major economic losses worldwide. The disease is caused by Newcastle Disease Virus (NDV) and early detection and identification of the viral strain is essential. Having knowledge of the NDV strain genotype that circulates in some regions would help in designing an effective vaccine to control the disease. In this regard, there is little information on NDV strain in chickens in mid Rift Valley and the central part of Ethiopia. Therefore, the purpose of this study was to detect and characterize NDV strain genotype from chickens in mid-Rift Valley and the central part of Ethiopia and test whether this NDV strain genotype matches the vaccine strain currently used in the study area.
Methods
A total of 98 samples: 78 (tracheal and cloacal) swabs from chicken pools of five and 20 tissue samples were collected. To detect NDV strain, conserved region of the virus Matrix (M) gene was amplified by qRT-PCR. To characterize NDV strain genotypes, M-gene positive samples were specifically re-amplified by conventional PCR targeting the Fusion (F) gene region and sequenced by Sanger method.
Results
13.26% of tested samples were positive for NDV strain in the study area with statistically significant difference (P<0.05) among the study sites. Further characterization of the F genes from NDV strain isolates by phylogenetic analysis indicated that one field isolate clustered with genotype VII whereas three of the isolates clustered to genotype I, II, and III. The isolate of the current NDV strain vaccine in use in the study area clustered with genotype II.
Conclusion
The current study indicates the existence of different NDV strain genotype from that of the vaccine strain currently used. Even though large-scale characterization of several isolates is required at national level, the current study laid baseline information for the existence of variations between field NDV strain genotype and vaccine strain currently used against ND in the country.
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