# Mitochondrial and Nuclear DNA Analyses of Rhipicephalus microplus from Mizoram, Northeast India: Insights into Genetic Diversity and Endosymbiont

## TL;DR

This study explores the genetic diversity and endosymbiont presence in Rhipicephalus microplus ticks from Mizoram, India, revealing new phylogenetic insights and regional population patterns.

## Contribution

The first molecular analysis of R. microplus in Mizoram, revealing novel phylogenetic and endosymbiont data from a strategically important region.

## Key findings

- R. microplus populations from India, Bangladesh, and Pakistan form a distinct matrilineal lineage within clade C.
- 24 COX1 haplotypes were identified globally, with only one found in India.
- Coxiella-like endosymbionts were detected in 95% of the analyzed tick specimens.

## Abstract

Background/Objectives: In this study, we conducted molecular identification of R.microplus and explored the genetic diversity of R. microplus for the first time in Mizoram, a Northeastern Hill (NEH) state of India bordering Myanmar. Methods: To assess genetic variation and evolutionary relationships, we employed phylogenetic analyses, genetic divergence metrics, and haplotype network construction based on mitochondrial (COX1 and 16S rDNA) and nuclear (ITS-2 and 18S rDNA) markers. Additionally, multivariate Principal Coordinate Analysis (PCoA) was used to visualize genetic differentiation among R. microplus populations. Results: Our analyses indicated that populations of R. microplus sensu lato from India, Bangladesh, and Pakistan form a closely related matrilineal lineage distinct from R. microplus sensu stricto, clustering within clade C of the COX1-based phylogeny. Globally, 24 COX1 haplotypes were recovered, with 1 haplotype identified in India. The Mizoram population exhibited a single 16S rDNA haplotype; however, intraspecific divergence was evident across India, with seven matrilineal haplotypes detected and nineteen globally. Further, five haplotypes were identified within R. microplus using the ITS-2 marker, while five haplotypes were observed within the Rhipicephalus genus using the 18S rDNA marker. Moreover, this study revealed the presence of Coxiella-like endosymbionts in 95% of the tick specimens analyzed. Conclusions: This study fills a critical knowledge gap by providing the first molecular documentation of tick diversity in Mizoram, a strategic region along the Indo–Myanmar border, and offers novel insights into the phylogeography and symbiotic associations of R. microplus and related tick taxa.