Vet World Vol.12 October-2019 Article-6
Research Article
Veterinary World, 12(10): 1546-1553
https://doi.org/10.14202/vetworld.2019.1546-1553
Identification of uncultured bacteria from abscesses of exotic pet animals using broad-range nested 16S rRNA polymerase chain reaction and Sanger sequencing
2. Private Consultant, 6 Gundaring Turn, Caning Vale, Western Australia 6155.
3. Exotic Pet Clinic, Faculty of Veterinary Medicine, Veterinary Teaching Hospital, Kasetsart University, Bangkok, Thailand.
4. Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.
5. Department of Infectious Diseases and Global Health, Cummings School of Veterinary Medicine, Tufts University, Boston, MA, USA.
6. Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, Bangkok, Thailand.
Background and Aim: This study demonstrated the usefulness of a broad-range polymerase chain reaction (PCR) with Sanger sequencing to identify the majority population of bacteria in abscesses from exotic pet animals.
Materials and Methods: This study performed a pilot investigation of abscesses from 20 clinical cases (17 rabbits, 2 hedgehogs, and 1 sugar glider) using standard culture methods for both aerobes and anaerobes and broad-range nested PCR targeting the 16S rRNA gene followed by the Sanger sequencing technique.
Results: The standard culture and PCR techniques detected bacteria in 9 and 17 of 20 samples, respectively. From the 17 sequencings of the 16S rRNA, 10 PCR products were found to be closely related with obligate anaerobes including Bacteroides spp., Fusobacterium spp., Prevotella spp. Phylogenetic analysis using the rpoB gene revealed that the species for the Bacteroides was thetaiotaomicron and for the Fusobacterium was varium and nucleatum. However, the amplification of the rpoB gene for the Prevotella spp. was unsuccessful. Correlations between the standard culture and PCR techniques were found in 9 (6 positive and 3 negative samples) of 20 samples. Eleven samples were discordant between the standard culture and PCR techniques which were composed of eight samples negative by culture but positive by PCR and three samples had different bacteria by the culture and PCR techniques.
Conclusion: According to this study, broad-range PCR combined with Sanger sequencing might be useful for the detection of dominant anaerobic bacteria in abscesses that were overlooked based on conventional bacterial culture. Keywords: anaerobic bacteria, abscesses, exotic pet animals, Sanger sequencing.
Keywords: anaerobic bacteria, abscesses, exotic pet animals, Sanger sequencing.
How to cite this article: Duangurai T, Siengsanan-Lamont J, Bumrungpun C, Kaewmongkol G, Areevijittrakul L, Sirinarumitr T, Fenwick SG, Kaewmongkol S (2019) Identification of uncultured bacteria from abscesses of exotic pet animals using broad-range nested 16S rRNA polymerase chain reaction and Sanger sequencing, Veterinary World, 12(10): 1546-1553.
Received: 30-05-2019 Accepted: 26-08-2019 Published online: 09-10-2019
Corresponding author: S. Kaewmongkol E-mail: cvtswt@ku.ac.th
DOI: 10.14202/vetworld.2019.1546-1553
Copyright: Duangurai, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.