Surface sterilization methods impact measures of internal microbial diversity in ticks
Abstract Background Ticks are obligate blood feeders transmitting major pathogens worldwide. Over the past few years, considerable research efforts have focused on the diversity, distribution and impact of gut and intracellular bacterial symbionts on tick development and tick-borne pathogen transmis...
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| Format: | Article |
| Language: | English |
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BMC
2019-05-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13071-019-3517-5 |
| _version_ | 1818482743121543168 |
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| author | Florian Binetruy Marlène Dupraz Marie Buysse Olivier Duron |
| author_facet | Florian Binetruy Marlène Dupraz Marie Buysse Olivier Duron |
| author_sort | Florian Binetruy |
| collection | DOAJ |
| description | Abstract Background Ticks are obligate blood feeders transmitting major pathogens worldwide. Over the past few years, considerable research efforts have focused on the diversity, distribution and impact of gut and intracellular bacterial symbionts on tick development and tick-borne pathogen transmission. The study of this internal microbiome requires the use of a sterilization method to remove external (i.e. cuticular) microbes present on the tick’s surface and to avoid any further contamination. Several sterilization methods exist, including ethanol- or bleach-based treatments that are both effective in killing microbes but with different potential effects on DNA denaturation. Methods We examined how these different sterilization methods impact the measure of internal microbial diversity hosted by the Cayenne tick Amblyomma cajennense (sensu stricto). Bacterial barcoding investigations based on 16S rRNA gene sequences were conducted on two batches of 50 individuals each: Ticks of the first batch were sterilized with bleach diluted at 1% and the second batch with 70% ethanol. Tick external microbiome was also determined from cuticle smearing and water samples used for tick washing. Results Bacterial barcoding investigations showed major differences between ethanol- and bleach-treated specimens. Both methods led to the detection of major intracellular bacteria associated with A. cajennense (s.s.) but ethanol-treated ticks always harbored a higher bacterial diversity than bleach-treated ticks. Further examinations of tick gut and tick external microbiome revealed that ethanol-based surface sterilization method is inefficient to eliminate the DNA of external bacteria. Conclusions We herein provide evidence that studies investigating the internal microbiome of ticks should consider bleach as the gold standard to efficiently remove cuticular bacterial DNA. Indeed, this method does not impact the internal bacterial diversity hosted by ticks and is thus a better method than the ethanol-based one for studying the internal microbiome. |
| first_indexed | 2024-12-10T11:51:10Z |
| format | Article |
| id | doaj.art-ed51c755c3e040a28094cb1a59a16529 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-12-10T11:51:10Z |
| publishDate | 2019-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-ed51c755c3e040a28094cb1a59a165292022-12-22T01:49:56ZengBMCParasites & Vectors1756-33052019-05-0112111010.1186/s13071-019-3517-5Surface sterilization methods impact measures of internal microbial diversity in ticksFlorian Binetruy0Marlène Dupraz1Marie Buysse2Olivier Duron3MIVEGEC (Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle), Centre National de la Recherche Scientifique (CNRS) - Institut pour la Recherche et le Développement (IRD) - Université de Montpellier (UM)MIVEGEC (Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle), Centre National de la Recherche Scientifique (CNRS) - Institut pour la Recherche et le Développement (IRD) - Université de Montpellier (UM)MIVEGEC (Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle), Centre National de la Recherche Scientifique (CNRS) - Institut pour la Recherche et le Développement (IRD) - Université de Montpellier (UM)MIVEGEC (Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle), Centre National de la Recherche Scientifique (CNRS) - Institut pour la Recherche et le Développement (IRD) - Université de Montpellier (UM)Abstract Background Ticks are obligate blood feeders transmitting major pathogens worldwide. Over the past few years, considerable research efforts have focused on the diversity, distribution and impact of gut and intracellular bacterial symbionts on tick development and tick-borne pathogen transmission. The study of this internal microbiome requires the use of a sterilization method to remove external (i.e. cuticular) microbes present on the tick’s surface and to avoid any further contamination. Several sterilization methods exist, including ethanol- or bleach-based treatments that are both effective in killing microbes but with different potential effects on DNA denaturation. Methods We examined how these different sterilization methods impact the measure of internal microbial diversity hosted by the Cayenne tick Amblyomma cajennense (sensu stricto). Bacterial barcoding investigations based on 16S rRNA gene sequences were conducted on two batches of 50 individuals each: Ticks of the first batch were sterilized with bleach diluted at 1% and the second batch with 70% ethanol. Tick external microbiome was also determined from cuticle smearing and water samples used for tick washing. Results Bacterial barcoding investigations showed major differences between ethanol- and bleach-treated specimens. Both methods led to the detection of major intracellular bacteria associated with A. cajennense (s.s.) but ethanol-treated ticks always harbored a higher bacterial diversity than bleach-treated ticks. Further examinations of tick gut and tick external microbiome revealed that ethanol-based surface sterilization method is inefficient to eliminate the DNA of external bacteria. Conclusions We herein provide evidence that studies investigating the internal microbiome of ticks should consider bleach as the gold standard to efficiently remove cuticular bacterial DNA. Indeed, this method does not impact the internal bacterial diversity hosted by ticks and is thus a better method than the ethanol-based one for studying the internal microbiome.http://link.springer.com/article/10.1186/s13071-019-3517-516S rRNABacterial communitiesTick microbiomeMetabarcodingAmblyomma |
| spellingShingle | Florian Binetruy Marlène Dupraz Marie Buysse Olivier Duron Surface sterilization methods impact measures of internal microbial diversity in ticks Parasites & Vectors 16S rRNA Bacterial communities Tick microbiome Metabarcoding Amblyomma |
| title | Surface sterilization methods impact measures of internal microbial diversity in ticks |
| title_full | Surface sterilization methods impact measures of internal microbial diversity in ticks |
| title_fullStr | Surface sterilization methods impact measures of internal microbial diversity in ticks |
| title_full_unstemmed | Surface sterilization methods impact measures of internal microbial diversity in ticks |
| title_short | Surface sterilization methods impact measures of internal microbial diversity in ticks |
| title_sort | surface sterilization methods impact measures of internal microbial diversity in ticks |
| topic | 16S rRNA Bacterial communities Tick microbiome Metabarcoding Amblyomma |
| url | http://link.springer.com/article/10.1186/s13071-019-3517-5 |
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