臺灣博碩士論文加值系統

Background. Wolbachia belongs to the order Rickettsiales and is considered a common intracellular bacteria within a variety of arthropods. As an endosymbiont, Wolbachia is not known to directly infect vertebrates and possesses an interesting ability to alter its host biology in diverse ways, such as suppression of vector population and interference of pathogen transmission. Previous studies have reported that Wolbachia plays a role in cytoplasmic incompatibility and is able to reduce dengue and Zika virus transmission in some mosquitoes. Hence, this symbiotic bacteria’s ability to manipulate its host may have potential applications in the vector control of tick-borne pathogens.

Aim. To determine the presence, prevalence, and phylogenetic analysis of symbiotic Wolbachia in Rhipicephalus sanguineus ticks collected from Kaohsiung, Taiwan

Methods. Ticks were collected from dogs throughout six districts of Kaohsiung, Taiwan. DNA samples were extracted from homogenized ticks using Qiagen DNeasy kit. Nested PCR assays were performed to detect wsp (Wolbachia surface protein) gene of Wolbachia supergroups A and B. Phylogenetic relationships were analyzed by Maximum likelihood method.

Results. In general, Wolbachia DNA was detected in 46% (463/1004) of the Rhipicephalus sanguineus ticks collected from dogs. The prevalence of infection was detected in males, females, and nymphs with an infection rate of 44%, 40% and 56%, respectively. Phylogenetic analysis also verifies these ticks as being either singly or superinfected with groups A and B of Wolbachia.

Conclusions. This study provides the first molecular evidence of Wolbachia infection in Rhipicephalus sanguineus ticks in Taiwan. Additionally, it is the first study to reveal the detection of Wolbachia in male Rhipicephalus sanguineus ticks and indicates the possible mechanism of vertical transmission within ticks. Based on the nested PCR assays, Rhipicephalus sanguineus ticks were found either singly or superinfected with groups A and B of Wolbachia.

Background. Wolbachia belongs to the order Rickettsiales and is considered a common intracellular bacteria within a variety of arthropods. As an endosymbiont, Wolbachia is not known to directly infect vertebrates and possesses an interesting ability to alter its host biology in diverse ways, such as suppression of vector population and interference of pathogen transmission. Previous studies have reported that Wolbachia plays a role in cytoplasmic incompatibility and is able to reduce dengue and Zika virus transmission in some mosquitoes. Hence, this symbiotic bacteria’s ability to manipulate its host may have potential applications in the vector control of tick-borne pathogens.

Aim. To determine the presence, prevalence, and phylogenetic analysis of symbiotic Wolbachia in Rhipicephalus sanguineus ticks collected from Kaohsiung, Taiwan

Methods. Ticks were collected from dogs throughout six districts of Kaohsiung, Taiwan. DNA samples were extracted from homogenized ticks using Qiagen DNeasy kit. Nested PCR assays were performed to detect wsp (Wolbachia surface protein) gene of Wolbachia supergroups A and B. Phylogenetic relationships were analyzed by Maximum likelihood method.

Results. In general, Wolbachia DNA was detected in 46% (463/1004) of the Rhipicephalus sanguineus ticks collected from dogs. The prevalence of infection was detected in males, females, and nymphs with an infection rate of 44%, 40% and 56%, respectively. Phylogenetic analysis also verifies these ticks as being either singly or superinfected with groups A and B of Wolbachia.

Conclusions. This study provides the first molecular evidence of Wolbachia infection in Rhipicephalus sanguineus ticks in Taiwan. Additionally, it is the first study to reveal the detection of Wolbachia in male Rhipicephalus sanguineus ticks and indicates the possible mechanism of vertical transmission within ticks. Based on the nested PCR assays, Rhipicephalus sanguineus ticks were found either singly or superinfected with groups A and B of Wolbachia.

Table of Contents.....i
List of Tables....ii
List of Figures.....ii
Abstract....iv-v

Chapter 1 Introduction....1-3
1.1. Background....1-2
1.1.1. Rhipicephalus sanguineus.....1-2
1.1.2. Wolbachia....2
1.2. Objectives....2-3
Chapter 2 Literature Review....4-7
2.1. Rhipicephalus sanguineus as a Vector of Diseases and Relevance to Taiwan.... 4
2.2. Wolbachia Based Biological Control....5
2.3. Wolbachia Detection in Ticks....6
2.4. Genetic Analysis of Wolbachia....6-7
Chapter 3 Materials and Methods....8-10
3.1. Rhipicephalus sanguineus Collection....8
3.2. DNA Extraction and PCR Analysis....8-9
3.2.1. DNA Extraction....8
3.2.2. DNA Amplification for Wolbachia....9
3.3. Sequencing and Phylogenetic Analysis....10
3.4. Statistical Analysis....10
Chapter 4 Results....11-12
4.1. Wolbachia Detection in Rhipicephalus sanguineus Ticks....11
4.2. Wolbachia Infection Rate....11
4.3. Phylogenetic Analysis of Wolbachia....12
Chapter 5 Discussions....13-16
References.....17-21
Appendix....32-33

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