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United States Department of Agriculture

Agricultural Research Service

Research Project: MINING THE GENOME OF RHIPICEPHALUS MICROPLUS TO DEVELOP NOVEL CONTROL TECHNOLOGY AND VACCINES

Location: Tick and Biting Fly Research

Title: Assessment of bacterial diversity in the cattle tick Rhipicephalus (Boophilus) microplus through tag-encoded pyrosequencing

Authors
item Andreotti, Renato -
item Perez De Leon, Adalberto
item Dowd, Scot
item Guerrero, Felix
item Bendele, Kylie
item Scoles, Glen

Submitted to: BMC Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 7, 2010
Publication Date: January 6, 2011
Citation: Andreotti, R., Perez De Leon, A.A., Dowd, S.E., Guerrero, F., Bendele, K.G., Scoles, G.A. 2011. Assessment of bacterial diversity in the cattle tick Rhipicephalus (Boophilus) microplus through tag-encoded pyrosequencing. BMC Microbiology. 11:article 6.

Interpretive Summary: Ticks are considered to be second only to mosquitoes as worldwide vectors of human diseases, but they are regarded as the most relevant vectors of disease-causing microorganisms in domestic and wild animals. The cattle tick, Rhipicephalus (Boophilus) microplus, hinders livestock production in tropical and subtropical parts of the world where it is endemic. The objective of this study was to characterize bacterial diversity in R. microplus using molecular methods that avoided the need to culture bacteria. Specifically, the 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) technique was used in this study. bTEFAP is a deoxyribonucleic acid (DNA) sequencing technique based on the detection of released pyrophosphate during DNA synthesis. DNA fragments were produced by copying information from the 16S ribosomal ribonucleic acid gene, which is highly conserved between different species of bacteria. bTEFAP was performed on adult males and females, eggs, and gut and ovary tissues from adult females derived from samples of R. microplus collected during outbreaks in southern Texas. Bioinformatic procedures were applied to the bTEFAP data prior to analysis. The total number of bacterial genera identified by tick sample comprised: 53 in adult males, 61 in adult females, 11 in gut tissue, 7 in ovarian tissue, and 54 in the eggs. Bacterial genera identified that are known to include species with pathogenic effects on vertebrate or invertebrate hosts included: Wolbachia, Staphylococcus, Coxiella, Corynebacterium, Enterococcus, Pseudomonas, Clostridium, Serratia, Borrelia, Streptococcus, Klebsiella, Moraxella, Enterobacter, Shigella and Escherichia. This work contributes qualitative and quantitative data for bacteria associated with the cattle tick R. microplus. Bacteria harbored by the cattle tick with the potential to be pathogenic to vertebrate hosts were identified. This report represents the first comprehensive survey of the bacteriome in the cattle tick using non-culture based molecular approaches.

Technical Abstract: Ticks are considered to be second only to mosquitoes as worldwide vectors of human diseases, but they are regarded as the most relevant vectors of disease-causing microorganisms in domestic and wild animals. The cattle tick, Rhipicephalus (Boophilus) microplus, hinders livestock production in tropical and subtropical parts of the world where it is endemic. The objective of this study was to apply the bacterial 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) technique to characterize bacterial diversity in R. microplus. Pyrosequencing was performed on adult males and females, eggs, and gut and ovary tissues from adult females derived from samples of R. microplus collected during outbreaks in southern Texas. Raw data from bTEFAP was screened and trimmed based upon quality scores and binned into individual sample collections. The total number of genera identified by tick sample comprised: 53 in adult males, 61 in adult females, 11 in gut tissue, 7 in ovarian tissue, and 54 in the eggs. Genera identified that are known to include species with pathogenic effects on vertebrate or invertebrate hosts included: Wolbachia, Staphylococcus, Coxiella, Corynebacterium, Enterococcus, Pseudomonas, Clostridium, Serratia, Borrelia, Streptococcus, Klebsiella, Moraxella, Enterobacter, Shigella and Escherichia. This work contributes qualitative and quantitative data for bacteria associated with the cattle tick R. microplus. Bacteria harbored by the cattle tick with the potential to be pathogenic to vertebrate hosts were identified. This report represents the first comprehensive survey of the bacteriome in the cattle tick using non-culture based molecular approaches.

Last Modified: 4/23/2014
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