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Viral and bacterial profiles in endemic influenza A virus infected swine herds using nanopore metagenomic sequencing on tracheobronchial swabs

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Abstract
Swine influenza A virus (swIAV) plays an important role in porcine respiratory infections. In addition to its ability to cause severe disease by itself, it is important in the multietiological porcine respiratory disease complex. Still, to date, no comprehensive diagnostics with which to study polymicrobial infections in detail have been offered. Hence, veterinary practitioners rely on monospecific and costly diagnostics, such as Reverse Transcription quantitative PCR (RT-qPCR), antigen detection, and serology. This prevents the proper understanding of the entire disease context, thereby hampering effective preventive and therapeutic actions. A new, nanopore-based, metagenomic diagnostic platform was applied to study viral and bacterial profiles across 4 age groups on 25 endemic swIAV-infected German farms with respiratory distress in the nursery. Farms were screened for swIAV using RT-qPCR on nasal and tracheobronchial swabs (TBS). TBS samples were pooled per age, prior to metagenomic characterization. The resulting data showed a correlation between the swIAV loads and the normalized reads, supporting a (semi-)quantitative interpretation of the metagenomic data. Interestingly, an in-depth characterization using beta diversity and PERMANOVA analyses allowed for the observation of an age-dependent interplay of known microbial agents. Also, lesser-known microbes, such as porcine polyoma, parainfluenza, and hemagglutinating encephalomyelitis viruses, were observed. Analyses of swIAV incidence and clinical signs showed differing microbial communities, highlighting age-specific observations of various microbes in porcine respiratory disease. In conclusion, nanopore metagenomics were shown to enable a panoramic view on viral and bacterial profiles as well as putative pathogen dynamics in endemic swIAV-infected herds. The results also highlighted the need for better insights into lesser studied agents that are potentially associated with porcine respiratory disease. IMPORTANCE To date, no comprehensive diagnostics for the study of polymicrobial infections that are associated with porcine respiratory disease have been offered. This precludes the proper understanding of the entire disease landscape, thereby hampering effective preventive and therapeutic actions. Compared to the often-costly diagnostic procedures that are applied for the diagnostics of porcine respiratory disease nowadays, a third-generation nanopore sequencing diagnostics workflow presents a cost-efficient and informative tool. This approach offers a panoramic view of microbial agents and contributes to the in-depth observation and characterization of viral and bacterial profiles within the respiratory disease context. While these data allow for the study of age-associated, swIAV-associated, and clinical symptom-associated observations, it also suggests that more effort should be put toward the investigation of coinfections and lesser-known pathogens (e.g., PHEV and PPIV), along with their potential roles in porcine respiratory disease. Overall, this approach will allow veterinary practitioners to tailor treatment and/or management changes on farms in a quicker, more complete, and cost-efficient way.
Keywords
Infectious Diseases, Cell Biology, Microbiology (medical), Genetics, General Immunology and Microbiology, Ecology, Physiology, coinfections, influenza A virus, nanopore sequencing, diagnostics, domestic pigs

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MLA
Vereecke, Nick, et al. “Viral and Bacterial Profiles in Endemic Influenza A Virus Infected Swine Herds Using Nanopore Metagenomic Sequencing on Tracheobronchial Swabs.” MICROBIOLOGY SPECTRUM, edited by Artem S. Rogovskyy, vol. 11, no. 2, American Society for Microbiology, 2023, doi:10.1128/spectrum.00098-23.
APA
Vereecke, N., Zwickl, S., Gumbert, S., Graaf, A., Harder, T., Ritzmann, M., … Stadler, J. (2023). Viral and bacterial profiles in endemic influenza A virus infected swine herds using nanopore metagenomic sequencing on tracheobronchial swabs. MICROBIOLOGY SPECTRUM, 11(2). https://doi.org/10.1128/spectrum.00098-23
Chicago author-date
Vereecke, Nick, Sophia Zwickl, Sophie Gumbert, Annika Graaf, Timm Harder, Mathias Ritzmann, Kathrin Lillie-Jaschniski, Sebastiaan Theuns, and Julia Stadler. 2023. “Viral and Bacterial Profiles in Endemic Influenza A Virus Infected Swine Herds Using Nanopore Metagenomic Sequencing on Tracheobronchial Swabs.” Edited by Artem S. Rogovskyy. MICROBIOLOGY SPECTRUM 11 (2). https://doi.org/10.1128/spectrum.00098-23.
Chicago author-date (all authors)
Vereecke, Nick, Sophia Zwickl, Sophie Gumbert, Annika Graaf, Timm Harder, Mathias Ritzmann, Kathrin Lillie-Jaschniski, Sebastiaan Theuns, and Julia Stadler. 2023. “Viral and Bacterial Profiles in Endemic Influenza A Virus Infected Swine Herds Using Nanopore Metagenomic Sequencing on Tracheobronchial Swabs.” Ed by. Artem S. Rogovskyy. MICROBIOLOGY SPECTRUM 11 (2). doi:10.1128/spectrum.00098-23.
Vancouver
1.
Vereecke N, Zwickl S, Gumbert S, Graaf A, Harder T, Ritzmann M, et al. Viral and bacterial profiles in endemic influenza A virus infected swine herds using nanopore metagenomic sequencing on tracheobronchial swabs. Rogovskyy AS, editor. MICROBIOLOGY SPECTRUM. 2023;11(2).
IEEE
[1]
N. Vereecke et al., “Viral and bacterial profiles in endemic influenza A virus infected swine herds using nanopore metagenomic sequencing on tracheobronchial swabs,” MICROBIOLOGY SPECTRUM, vol. 11, no. 2, 2023.
@article{01GVJ595C66MFEZWHGKDR8X550,
  abstract     = {{Swine influenza A virus (swIAV) plays an important role in porcine respiratory infections. In addition to its ability to cause severe disease by itself, it is important in the multietiological porcine respiratory disease complex. Still, to date, no comprehensive diagnostics with which to study polymicrobial infections in detail have been offered. Hence, veterinary practitioners rely on monospecific and costly diagnostics, such as Reverse Transcription quantitative PCR (RT-qPCR), antigen detection, and serology. This prevents the proper understanding of the entire disease context, thereby hampering effective preventive and therapeutic actions. A new, nanopore-based, metagenomic diagnostic platform was applied to study viral and bacterial profiles across 4 age groups on 25 endemic swIAV-infected German farms with respiratory distress in the nursery. Farms were screened for swIAV using RT-qPCR on nasal and tracheobronchial swabs (TBS). TBS samples were pooled per age, prior to metagenomic characterization. The resulting data showed a correlation between the swIAV loads and the normalized reads, supporting a (semi-)quantitative interpretation of the metagenomic data. Interestingly, an in-depth characterization using beta diversity and PERMANOVA analyses allowed for the observation of an age-dependent interplay of known microbial agents. Also, lesser-known microbes, such as porcine polyoma, parainfluenza, and hemagglutinating encephalomyelitis viruses, were observed. Analyses of swIAV incidence and clinical signs showed differing microbial communities, highlighting age-specific observations of various microbes in porcine respiratory disease. In conclusion, nanopore metagenomics were shown to enable a panoramic view on viral and bacterial profiles as well as putative pathogen dynamics in endemic swIAV-infected herds. The results also highlighted the need for better insights into lesser studied agents that are potentially associated with porcine respiratory disease.

IMPORTANCE To date, no comprehensive diagnostics for the study of polymicrobial infections that are associated with porcine respiratory disease have been offered. This precludes the proper understanding of the entire disease landscape, thereby hampering effective preventive and therapeutic actions. Compared to the often-costly diagnostic procedures that are applied for the diagnostics of porcine respiratory disease nowadays, a third-generation nanopore sequencing diagnostics workflow presents a cost-efficient and informative tool. This approach offers a panoramic view of microbial agents and contributes to the in-depth observation and characterization of viral and bacterial profiles within the respiratory disease context. While these data allow for the study of age-associated, swIAV-associated, and clinical symptom-associated observations, it also suggests that more effort should be put toward the investigation of coinfections and lesser-known pathogens (e.g., PHEV and PPIV), along with their potential roles in porcine respiratory disease. Overall, this approach will allow veterinary practitioners to tailor treatment and/or management changes on farms in a quicker, more complete, and cost-efficient way.}},
  articleno    = {{e00098-23}},
  author       = {{Vereecke, Nick and Zwickl, Sophia and Gumbert, Sophie and Graaf, Annika and Harder, Timm and Ritzmann, Mathias and Lillie-Jaschniski, Kathrin and Theuns, Sebastiaan and Stadler, Julia}},
  editor       = {{Rogovskyy, Artem S.}},
  issn         = {{2165-0497}},
  journal      = {{MICROBIOLOGY SPECTRUM}},
  keywords     = {{Infectious Diseases,Cell Biology,Microbiology (medical),Genetics,General Immunology and Microbiology,Ecology,Physiology,coinfections,influenza A virus,nanopore sequencing,diagnostics,domestic pigs}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{20}},
  publisher    = {{American Society for Microbiology}},
  title        = {{Viral and bacterial profiles in endemic influenza A virus infected swine herds using nanopore metagenomic sequencing on tracheobronchial swabs}},
  url          = {{http://doi.org/10.1128/spectrum.00098-23}},
  volume       = {{11}},
  year         = {{2023}},
}

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