Reptile-associated Borrelia species in the goanna tick (Bothriocroton undatum) from Sydney, Australia
Abstract Background Knowledge on the capacity of Australian ticks to carry Borrelia species is currently limited or missing. To evaluate the potential of ticks to carry bacterial pathogens and their DNA, it is imperative to have a robust workflow that maximises recovery of bacterial DNA within ticks...
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| Format: | Article |
| Language: | English |
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BMC
2017-12-01
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| Series: | Parasites & Vectors |
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| Online Access: | http://link.springer.com/article/10.1186/s13071-017-2579-5 |
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| author | Jessica L. Panetta Radek Šíma Nichola E. D. Calvani Ondřej Hajdušek Shona Chandra Jessica Panuccio Jan Šlapeta |
| author_facet | Jessica L. Panetta Radek Šíma Nichola E. D. Calvani Ondřej Hajdušek Shona Chandra Jessica Panuccio Jan Šlapeta |
| author_sort | Jessica L. Panetta |
| collection | DOAJ |
| description | Abstract Background Knowledge on the capacity of Australian ticks to carry Borrelia species is currently limited or missing. To evaluate the potential of ticks to carry bacterial pathogens and their DNA, it is imperative to have a robust workflow that maximises recovery of bacterial DNA within ticks in order to enable accurate identification. By exploiting the bilateral anatomical symmetry of ticks, we were able to directly compare two DNA extraction methods for 16S rRNA gene diversity profiling and pathogen detection. We aimed to assess which combination of DNA extraction and 16S rRNA hypervariable region enables identification of the greatest bacterial diversity, whilst minimising bias, and providing the greatest capacity for the identification of Borrelia spp. Results We collected Australian endemic ticks (Bothriocroton undatum), isolated DNA from equal tick halves using two commercial DNA extraction methods and sequenced samples using V1-V3 and V3-V4 16S rRNA gene diversity profiling assays. Two distinct Borrelia spp. operational taxonomic units (OTUs) were detected using the V1-V3 16S rRNA hypervariable region and matching Borrelia spp. sequences were obtained using a conventional nested-PCR. The tick 16S rRNA gene diversity profile was dominated by Rickettsia spp. (98–99%), while the remaining OTUs belonged to Proteobacteria (51–81%), Actinobacteria (6–30%) and Firmicutes (2–7%). Multiple comparisons tests demonstrated biases in each of the DNA extraction kits towards different bacterial taxa. Conclusions Two distinct Borrelia species belonging to the reptile-associated Borrelia group were identified. Our results show that the method of DNA extraction can promote bias in the final microbiota identified. We determined an optimal DNA extraction method and 16S rRNA gene diversity profile assay that maximises detection of Borrelia species. |
| first_indexed | 2024-12-23T13:17:58Z |
| format | Article |
| id | doaj.art-70dad083a4e642c1884f2ef2f0569cc5 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-12-23T13:17:58Z |
| publishDate | 2017-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-70dad083a4e642c1884f2ef2f0569cc52022-12-21T17:45:33ZengBMCParasites & Vectors1756-33052017-12-0110111310.1186/s13071-017-2579-5Reptile-associated Borrelia species in the goanna tick (Bothriocroton undatum) from Sydney, AustraliaJessica L. Panetta0Radek Šíma1Nichola E. D. Calvani2Ondřej Hajdušek3Shona Chandra4Jessica Panuccio5Jan Šlapeta6Sydney School of Veterinary Science, Faculty of Science, University of SydneyInstitute of Parasitology, Biology Centre of the Czech Academy of SciencesSydney School of Veterinary Science, Faculty of Science, University of SydneyInstitute of Parasitology, Biology Centre of the Czech Academy of SciencesSydney School of Veterinary Science, Faculty of Science, University of SydneySydney School of Veterinary Science, Faculty of Science, University of SydneySydney School of Veterinary Science, Faculty of Science, University of SydneyAbstract Background Knowledge on the capacity of Australian ticks to carry Borrelia species is currently limited or missing. To evaluate the potential of ticks to carry bacterial pathogens and their DNA, it is imperative to have a robust workflow that maximises recovery of bacterial DNA within ticks in order to enable accurate identification. By exploiting the bilateral anatomical symmetry of ticks, we were able to directly compare two DNA extraction methods for 16S rRNA gene diversity profiling and pathogen detection. We aimed to assess which combination of DNA extraction and 16S rRNA hypervariable region enables identification of the greatest bacterial diversity, whilst minimising bias, and providing the greatest capacity for the identification of Borrelia spp. Results We collected Australian endemic ticks (Bothriocroton undatum), isolated DNA from equal tick halves using two commercial DNA extraction methods and sequenced samples using V1-V3 and V3-V4 16S rRNA gene diversity profiling assays. Two distinct Borrelia spp. operational taxonomic units (OTUs) were detected using the V1-V3 16S rRNA hypervariable region and matching Borrelia spp. sequences were obtained using a conventional nested-PCR. The tick 16S rRNA gene diversity profile was dominated by Rickettsia spp. (98–99%), while the remaining OTUs belonged to Proteobacteria (51–81%), Actinobacteria (6–30%) and Firmicutes (2–7%). Multiple comparisons tests demonstrated biases in each of the DNA extraction kits towards different bacterial taxa. Conclusions Two distinct Borrelia species belonging to the reptile-associated Borrelia group were identified. Our results show that the method of DNA extraction can promote bias in the final microbiota identified. We determined an optimal DNA extraction method and 16S rRNA gene diversity profile assay that maximises detection of Borrelia species.http://link.springer.com/article/10.1186/s13071-017-2579-5Bothriocroton undatumBorreliaGoanna tickDNA extractionMiSeqIllumina |
| spellingShingle | Jessica L. Panetta Radek Šíma Nichola E. D. Calvani Ondřej Hajdušek Shona Chandra Jessica Panuccio Jan Šlapeta Reptile-associated Borrelia species in the goanna tick (Bothriocroton undatum) from Sydney, Australia Parasites & Vectors Bothriocroton undatum Borrelia Goanna tick DNA extraction MiSeq Illumina |
| title | Reptile-associated Borrelia species in the goanna tick (Bothriocroton undatum) from Sydney, Australia |
| title_full | Reptile-associated Borrelia species in the goanna tick (Bothriocroton undatum) from Sydney, Australia |
| title_fullStr | Reptile-associated Borrelia species in the goanna tick (Bothriocroton undatum) from Sydney, Australia |
| title_full_unstemmed | Reptile-associated Borrelia species in the goanna tick (Bothriocroton undatum) from Sydney, Australia |
| title_short | Reptile-associated Borrelia species in the goanna tick (Bothriocroton undatum) from Sydney, Australia |
| title_sort | reptile associated borrelia species in the goanna tick bothriocroton undatum from sydney australia |
| topic | Bothriocroton undatum Borrelia Goanna tick DNA extraction MiSeq Illumina |
| url | http://link.springer.com/article/10.1186/s13071-017-2579-5 |
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