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Genetic Structure of Amblyomma Cajennense (Acari: Ixodidae) Populations Based on Mitochondrial Gene Sequences

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Georgia Southern University Digital Commons@Georgia Southern Electronic Theses and Dissertations Graduate Studies, Jack N. Averitt College of Spring 2009 Genetic Structure of Amblyomma Cajennense (Acari: Ixodidae) Populations Based on Mitochondrial Gene Sequences Erica Janelle Burkman Follow this and additional works at: https://digitalcommons.georgiasouthern.edu/etd Recommended Citation Burkman, Erica Janelle, "Genetic Structure of Amblyomma Cajennense (Acari: Ixodidae) Populations Based on Mitochondrial Gene Sequences" (2009). Electronic Theses and Dissertations. 704. https://digitalcommons.georgiasouthern.edu/etd/704 This thesis (open access) is brought to you for free and open access by the Graduate Studies, Jack N. Averitt College of at Digital Commons@Georgia Southern. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of Digital Commons@Georgia Southern. For more information, please contact [email protected]. GENETIC STRUCTURE OF AMBLYOMMA CAJENNENSE (ACARI: IXODIDAE) POPULATIONS BASED ON MITOCHONDRIAL GENE SEQUENCES by ERICA JANELLE BURKMAN (Under the Direction of Dr. Lorenza Beati) ABSTRACT Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) is a common tick species that has a large geographic distribution from the southern regions of the United States (Texas), to the Caribbean Islands, Central, and South America. This tick is a vector of the agent of Brazilian spotted fever, an often fatal disease in South America. Throughout its geographic range, populations of A. cajennense have shown differences in ecological adaptation while feeding on a variety of hosts ranging from livestock, birds, and humans. In order to examine the taxonomic status and phylogeographic evolution of this species, we analyzed mitochondrial 12S rDNA, control region (d-loop), and cytochrome oxidase II gene sequences of A. cajennense specimens collected in eight different localities. The results showed that our samples are grouped in five strongly supported monophyletic lineages, each corresponding to geographically or ecologically distinct populations. The strong phylogenetic structure indicates that A. cajennense may actually be a species complex in need of thorough morphological reassessment. INDEX WORDS: Amblyomma cajennense, 12S rDNA, Control region (d-loop), Cytochrome oxidase II, Brazilian spotted fever, Cryptic species, Phylogeography, Genetic structure GENETIC STRUCTURE OF AMBLYOMMA CAJENNENSE (ACARI: IXODIDAE) POPULATIONS BASED ON MITOCHONDRIAL GENE SEQUENCES by ERICA JANELLE BURKMAN B.S., Florida International University, 2002 A Thesis Submitted to the Graduate Faculty of Georgia Southern University in Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE STATESBORO, GEORGIA 2009 2 © 2009 Erica Janelle Burkman All Rights Reserved 3 GENETIC STRUCTURE OF AMBLYOMMA CAJENNENSE (ACARI: IXODIDAE) POPULATIONS BASED ON MITOCHONDRIAL GENE SEQUENCES by ERICA JANELLE BURKMAN Major Professor: Lorenza Beati Committee: Lorenza Beati Lance Durden William Irby Electronic Version Approved: May 2009 4 TABLE OF CONTENTS Page LIST OF TABLES...............................................................................................................7 LIST OF FIGURES .............................................................................................................8 CHAPTER 1 INTRODUCTION .............................................................................................9 Taxonomic information.................................................................................9 Synonymy list................................................................................................9 Geographic distribution...............................................................................10 Host Associations........................................................................................11 Economic impact.........................................................................................11 Ticks and Population genetic studies ..........................................................12 Main Hypothesis..........................................................................................13 Purpose of study ..........................................................................................13 2 MATERIALS AND METHODS.....................................................................14 Sample Collection .......................................................................................14 DNA Extraction...........................................................................................15 PCR Analysis ..............................................................................................16 DNA Purification ........................................................................................18 Sequencing ..................................................................................................18 Assembly .....................................................................................................18 Alignment....................................................................................................18 5 3 RESULTS ........................................................................................................20 Phylogenetic analysis: 12S rDNA..............................................................20 D-loop..........................................................................................................24 COII.............................................................................................................28 Concatenated analysis .................................................................................32 4 CONCLUSION................................................................................................35 Sampling......................................................................................................35 Choice of gene sequences............................................................................35 Phylogenetic analyses..................................................................................37 Future studies ..............................................................................................39 REFERENCES ..................................................................................................................42 APPENDICES ...................................................................................................................47 A Location and collectors of samples used in this study.....................................47 B Results of DNA extraction from samples ........................................................48 C A. cajennense haplotypes obtained from samples............................................55 6 LIST OF TABLES Page Table 1: Amblyomma cajennense were collected from the following countries and the biomes of the sampled areas..............................................................................15 Table 2: Lists the gene position of the PCR amplification mixture and the primers used....................................................................................................................17 Table 3: PCR mixtures and programs for 12S, Control Region (dloop) and Cytochrome Oxidase II ..........................................................................................................17 Table 4: Maximum likelihood distances (%) between 12S rDNA sequences...................21 Table 5: Maximum likelihood distances (%) between D-loop sequences.........................25 Table 6: Maximum likelihood distances (%) between COII sequences............................28 7 LIST OF FIGURES Page Figure 1: Map of known distribution of A. cajennense ....................................................10 Figure 2: Geographic range of A. cajennense in South American biomes ........................14 Figure 3: Strict consensus tree of A. cajennense 12S rDNA gene sequences....................22 Figure 4: Bayesian phylogeny of A. cajennense 12S rDNA gene sequences....................23 Figure 5: Strict consensus tree of A. cajennense D-loop gene sequences .........................26 Figure 6: Bayesian phylogeny of A. cajennense D-loop gene sequences..........................27 Figure 7: Strict consensus tree of A. cajennense COII gene sequences.............................30 Figure 8: Bayesian phylogeny of A. cajennense COII gene sequences.............................31 Figure 9: Concatenated strict consensus of A. cajennense gene sequences.......................33 Figure 10: Concatenated Bayesian phylogeny of A. cajennense gene sequences .............34 Figure 11: Mitochondrial genome of Rhipicephalus sanguineus ......................................36 Figure 12: Geographic clades compared to concatenated Bayesian phylogeny................41 8 CHAPTER 1 INTRODUCTION Taxonomic information Amblyomma cajennense (Fabricius, 1787), commonly known as the Cayenne tick, is a common tick species infesting livestock and a number of other vertebrates including humans and birds (Borges et al., 2002; Oliviera et al., 2003; Lopes et al., 1998; Rojas, Marini and Coutinho, 1999). This tick has a wide distribution, ranging from the southern regions of the United States to the Neotropics. Because this tick is so widespread, taxonomists have been studying it since the late 18th Century and several names have been applied to it as shown below Synonymy list Acarus cajennensis Fabricius,
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