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We report the complete genome sequences of two non-typical Avibacterium paragallinarum (AP) strains isolated from chickens in the absence of clinical signs. The availability of these genomes can aid scientists in improving current diagnostics and increase our understanding of AP epidemiology and pathogenicity in chickens.
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Respiratory infections caused by Ornithobacterium rhinotrachealis (ORT) and Pasteurella multocida (PM) bacteria are significant threats to the poultry industry by causing economic losses and welfare issues. Due to characterization difficulties and underutilization of epidemiological tools, description of the spatio-temporal spread of these diseases in the field is limited. The objectives of this retrospective observational cross-sectional study were to (a) investigate the existence of space-time clusters (hotspots); and (b) investigate the association between genetic similarity and spatial proximity for both pathogens using molecular typing and a recently developed Core-Genome Multilocus Sequencing Typing (cgMLST) scheme. ORT (n = 103) and PM (n = 69) isolates from confirmed disease outbreaks from one commercial company between 2013 and 2021 were obtained from a veterinary diagnostic laboratory, characterized using a cgMLST scheme and visualized using a minimum spanning tree. Spatio-temporal cluster analysis using SaTScanTM and a Spearman's rank correlation were performed to investigate clustering and any association between allelic diversity and geospatial distance. The cgMLST sequencing revealed three allelic clusters for ORT and thirteen clusters for PM. The spatio-temporal analysis revealed two significant clusters for PM, one with a 259.3 km cluster containing six cases between May and July 2018 and a 9 km cluster containing five cases between February 2019 and February 2021. No spatio-temporal clusters were found for ORT. A weak negative correlation between allelic diversity and geospatial distance was observed for ORT (r = -0.04, p < 0.01) and a weak positive correlation was observed for PM (r = 0.11, p < 0.01). This study revealed regional spatio-temporal clusters for PM in commercial turkey sites between 2018 and 2021 and provided additional insight into bacterial strain subgroups and the geographical spread of ORT and PM over time.
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Pasteurella multocida is one of the major causes of mass mortalities in wild birds. Here, we report the complete genome sequences of two P. multocida isolates from wild populations of two endangered seabird species, the Indian yellow-nosed albatrosses (Thalassarche carteri) and the northern rockhopper penguins (Eudyptes moseleyi).
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Malacoplasma iowae, previously known as "Mycoplasma iowae," is associated with embryo mortality, reduced hatchability, and leg abnormalities in turkeys, leading to considerable economic losses. Here, we report the complete and annotated genome sequence of Malacoplasma iowae type strain 695.
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Ornithobacterium rhinotracheale has been associated with respiratory disease in poultry, particularly turkeys, leading to significant economic losses. However, O. rhinotracheale is poorly studied, and a very limited number of complete genomes are available. Here, we report the complete genome sequences of three O. rhinotracheale strains, generated using a Nanopore-Illumina hybrid assembly approach.
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The authors would like to make the following corrections to this paper [...].
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Ornithobacterium rhinotracheale (ORT) has been associated with poultry respiratory disease worldwide. The organism is fastidious and isolation is challenging. One TaqMan real-time PCR (qPCR) assay has been developed for the detection of ORT. However, during validating the ORT qPCR, the assay performance was suboptimal. During the in silico evaluation, deviations from the basic parameters for primers and probes designs (e.g., presence of stable undesirable primer-dimers) were observed. The suboptimal design led to low efficiency and low sensitivity of the assay. Initially, modification on the probe was carried out to improve the performance of the assay. However, the assay's performance (efficiency and sensitivity) was still suboptimal. In this manuscript, we describe the development of a new qPCR assay and the comparison of its performance with the currently available assay. A highly efficient, sensitive, and specific qPCR assay was developed with approximately 1000-folds reduction in the limit of detection (from 3 × 106 plasmid DNA copies/mL to 1 × 103 plasmid DNA copies/mL). Additionally, the efficiency of the new assay (E = 98.70%) was significantly better than the current assay (E = 73.18%). The newly developed assay is an improved diagnostic tool for the sensitive and efficient diagnosis of ORT from clinical samples.
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Bordetella avium (BA) is one of many pathogens that cause respiratory diseases in turkeys. However, other bacterial species can easily overgrow it during isolation attempts. This makes confirming the diagnosis of BA as the causative agent of turkey coryza more difficult. Currently, there are two PCR assays for the molecular detection of BA. One is conventional gel-based PCR and the other is TaqMan real-time PCR (qPCR) assay. However, multiple pitfalls were detected in both assays regarding their specificity, sensitivity, and efficiency, which limits their utility as diagnostic tools. In this study, we developed and validated two TaqMan qPCR assays and compared their performance to the currently available TaqMan qPCR. The two assays were able to correctly identify all BA isolates and showed negative results against a wide range of different microorganisms. The two assays were found to have high efficiency with a detection limit of approximately 1 × 103 plasmid DNA Copies/mL with high repeatability and reproducibility. In comparison to the currently available TaqMan qPCR assay, the newly developed assays showed significantly higher PCR efficiencies due to superior primers and probes design. The new assays can serve as a reliable tool for the sensitive, specific, and efficient diagnosis of BA.
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BACKGROUND AND AIM: Poultry infections with H9N2 avian influenza viruses (AIVs) are endemic in Egypt. This study determined the genetic changes in the sequences of H9N2 AIVs isolated from chicken and quails in Egypt, including determining genetic reassortment and detecting the main genetic changes in hemagglutinin (HA) and neuraminidase (NA) genes. MATERIALS AND METHODS: Swab samples were collected from chicken and quails, examined through reverse transcription-polymerase chain reaction, and AIVs from positive samples were isolated in embryonated chicken eggs. Complete genome sequencing and phylogenetic analyses were conducted for two H9N2 AIV isolates, and sequences of HA and NA gene segments were analyzed in another two isolates. RESULTS: A novel reassortant virus was identified from a commercial chicken flock (A/chicken/Egypt/374V/2016) and quails from a live bird market (A/quail/Egypt/1253V/2016). The reassortant viruses acquired four genome segments from the classic Egyptian H9N2 viruses (HA, NA, NP, and M) and four segments from Eurasian AIVs (PB2, PB1, PA, and NS). Many genetic changes have been demonstrated in HA and NA genes. The isolated novel reassortant H9N2 virus from quails showed amino acid mutations in the antigenic sites on the globular head of the mature HA monomer matched with the parent Egyptian H9N2 virus. CONCLUSION: This work described the genetic characterization of a novel reassortment of the H9N2 virus in Egypt. The emergence of new reassorted AIV viruses and genome variability raises the concern of an influenza pandemic with zoonotic potentials.
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Avian infectious bronchitis virus (AvIBV) is the causative agent of a highly contagious respiratory disease in chickens which results in significant economic losses in the poultry industry. Here, we report a near-complete genome sequence of the strain, designated IA1162/2020, identified in tracheal swabs from chickens in Iowa in 2020.
