Cargando…
Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora
Respiratory disease is responsible for dramatic population declines in bighorn sheep (Ovis canadensis), and respiratory pathogen diagnostics contribute to the management of bighorn populations. To create a comprehensive and consistent approach to bighorn sheep respiratory diagnostics, we created a c...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10586700/ https://www.ncbi.nlm.nih.gov/pubmed/37856492 http://dx.doi.org/10.1371/journal.pone.0293062 |
_version_ | 1785123202224095232 |
---|---|
author | Fox, Karen A. MacGlover, Christopher A. W. Blecha, Kevin A. Stenglein, Mark D. |
author_facet | Fox, Karen A. MacGlover, Christopher A. W. Blecha, Kevin A. Stenglein, Mark D. |
author_sort | Fox, Karen A. |
collection | PubMed |
description | Respiratory disease is responsible for dramatic population declines in bighorn sheep (Ovis canadensis), and respiratory pathogen diagnostics contribute to the management of bighorn populations. To create a comprehensive and consistent approach to bighorn sheep respiratory diagnostics, we created a culture-independent assay to detect and strain type Mannheimia haemolytica, Bibersteinia trehalosi, Pasteurella multocida, and Mycoplasma ovipneumoniae. The assay also detects and characterizes the Pasteurellaceae leukotoxin A gene, and broadly assesses the bacterial composition of each sample based on 16S rRNA sequences. The assay is based on a three-step approach: 1) Multiplex PCR to amplify targets including eight loci for each bacterial species, the Pasteurellaceae lktA gene, and the 16S rRNA gene 2) Library preparation, barcoding, and short-read Illumina sequencing to determine the genetic sequences of each target, and 3) Bioinformatics in the form of automated software to analyze genetic sequences. The assay was designed to assess shared pathogens between domestic and bighorn sheep, but could be useful for many applications in bighorn sheep respiratory disease research and management. |
format | Online Article Text |
id | pubmed-10586700 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-105867002023-10-20 Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora Fox, Karen A. MacGlover, Christopher A. W. Blecha, Kevin A. Stenglein, Mark D. PLoS One Research Article Respiratory disease is responsible for dramatic population declines in bighorn sheep (Ovis canadensis), and respiratory pathogen diagnostics contribute to the management of bighorn populations. To create a comprehensive and consistent approach to bighorn sheep respiratory diagnostics, we created a culture-independent assay to detect and strain type Mannheimia haemolytica, Bibersteinia trehalosi, Pasteurella multocida, and Mycoplasma ovipneumoniae. The assay also detects and characterizes the Pasteurellaceae leukotoxin A gene, and broadly assesses the bacterial composition of each sample based on 16S rRNA sequences. The assay is based on a three-step approach: 1) Multiplex PCR to amplify targets including eight loci for each bacterial species, the Pasteurellaceae lktA gene, and the 16S rRNA gene 2) Library preparation, barcoding, and short-read Illumina sequencing to determine the genetic sequences of each target, and 3) Bioinformatics in the form of automated software to analyze genetic sequences. The assay was designed to assess shared pathogens between domestic and bighorn sheep, but could be useful for many applications in bighorn sheep respiratory disease research and management. Public Library of Science 2023-10-19 /pmc/articles/PMC10586700/ /pubmed/37856492 http://dx.doi.org/10.1371/journal.pone.0293062 Text en © 2023 Fox et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Fox, Karen A. MacGlover, Christopher A. W. Blecha, Kevin A. Stenglein, Mark D. Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora |
title | Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora |
title_full | Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora |
title_fullStr | Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora |
title_full_unstemmed | Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora |
title_short | Assessing shared respiratory pathogens between domestic (Ovis aries) and bighorn (Ovis canadensis) sheep; methods for multiplex PCR, amplicon sequencing, and bioinformatics to characterize respiratory flora |
title_sort | assessing shared respiratory pathogens between domestic (ovis aries) and bighorn (ovis canadensis) sheep; methods for multiplex pcr, amplicon sequencing, and bioinformatics to characterize respiratory flora |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10586700/ https://www.ncbi.nlm.nih.gov/pubmed/37856492 http://dx.doi.org/10.1371/journal.pone.0293062 |
work_keys_str_mv | AT foxkarena assessingsharedrespiratorypathogensbetweendomesticovisariesandbighornoviscanadensissheepmethodsformultiplexpcrampliconsequencingandbioinformaticstocharacterizerespiratoryflora AT macgloverchristopheraw assessingsharedrespiratorypathogensbetweendomesticovisariesandbighornoviscanadensissheepmethodsformultiplexpcrampliconsequencingandbioinformaticstocharacterizerespiratoryflora AT blechakevina assessingsharedrespiratorypathogensbetweendomesticovisariesandbighornoviscanadensissheepmethodsformultiplexpcrampliconsequencingandbioinformaticstocharacterizerespiratoryflora AT stengleinmarkd assessingsharedrespiratorypathogensbetweendomesticovisariesandbighornoviscanadensissheepmethodsformultiplexpcrampliconsequencingandbioinformaticstocharacterizerespiratoryflora |