Vol.:(0123456789) 1 3 https://doi.org/10.1007/s00436-022-07596-3 ARTHROPODS AND MEDICAL ENTOMOLOGY - ORIGINAL PAPER Epidemiologic profile of hard ticks and molecular characterization of Rhipicephalus microplus infesting cattle in central part of Khyber Pakhtunkhwa, Pakistan Samia Sultan 1  · Jehan Zeb 1,6  · Sultan Ayaz 2  · Sadeeq Ur Rehman 2  · Sanaullah khan 3  · Mubashir Hussain 4  · Haytham Senbill 5  · Sabir Husain 6  · Olivier Andre Sparagano 6 Received: 13 December 2021 / Accepted: 4 July 2022 © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022 Abstract Tick infestation is a major public and animal health concern causing significant financial losses, especially in tropical and subtropical regions of the world. This study aimed at investigating the epidemiologic profile of ticks infesting cattle and molecular identification of R. microplus in the centrally ignored part of Khyber Pakhtunkhwa, Pakistan. A total of 600 cattle from 20 farms were exam- ined for the tick infestation, among them 358 (59.7%) cattle were infested with ticks. A total of 2118 nymph, larvae and adult tick stages were collected and morphologically identified followed by molecular confirmation of Rhipicephalus microplus. Host-based demographic and ecological parameter analysis revealed significantly higher tick infestation in adult, female, exotic, freely grazing, and with irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, gender, breed, acaricides use, and feeding method were significantly (P < 0.05) associated, whereas multivariate analysis revealed only host breed and feeding method were potential risk factors (P < 0.05) for tick infestation. Microscopy-based examination identified four different species of ticks including R. microplus (44.5%), Hyalomma anatolicum (38.5%), and Hyalomma marginatum (10.5%) and Hyalomma excavatum (6.5%). Tick infestation pattern showed that 55.9% of cattle was found co-infested with R. microplus and H. anatolicum followed by R. microplus and H. anatolicum and H. marginatum (29.3%) then R. microplus, H. anatolicum, H. marginatum, and H. excavatum (11.2%). Sequencing of the second internal transcribed spacer (ITS2-) and 16S rRNA gene fragments also confirmed the molecular identification of Rhipicephalus microplus. Phylogenetic analysis of ITS-2 revealed all sequences clustered in single clade of the R. microplus while the 16S rRNA nucleotide sequences showed that R. microplus in this study was clustered together in clade A along with other isolates from Pakistan, China, and India. The high tick infestation suggests the need for designing strategic and integrated control measures for ticks in order to ensure good health of domestic animals in this region of Pakistan. Keywords Tick infestation · R. microplus · Cattle · Microscopy · 16S rRNA Introduction Pakistan is an agricultural country, contributing 20.9% of the country GDP (gross domestic product) and 43.4% employer workforce according to 2013/2014 Pakistan Livestock Census (Ministry of Finance | Government of Pakistan 2013–2014). According to a recent survey, livestock herd comprises more than 213 million animals. This constitutes 42.4 million Riv- erine buffaloes and 51.5 million cattle (Pakistan Economic Survey 2020–2021), which can fulfill more than 97% of the country’s gross milk demand (Hussain et al. 2021). The FAO estimates that one-third of the developing world’s milk pro- duction is derived from dairy farms in India and Pakistan (FAO 2018). In Pakistan, smallholder dairy farms face huge challenges of various vector-borne diseases particularly ticks Section Editor: Leonhard Schnittger. Highlights 1. The study shows higher tick prevalence in cattle population in central part of Khyber Pakhtunkhwa, Pakistan. 2. Morphological identification showed the presence of four different species of ticks including R. microplus, Hyalomma anatolicum, Hyalomma marginatum, and Hyalomma excavatum. 3. Grazing and cattle breed are the significant risk factors associated with higher tick infestation. 4. 16S rRNA is a standard genetic marker in differentiating R. microplus genetic assemblages from a wide range of biogeographical regions. 5. ITS-2-based phylogenetic analysis revealed a single clade of R. microplus while based on the 16S rRNA nucleotide sequencing; R. microplus was assigned to clade A in the present study. Extended author information available on the last page of the article / Published online: 15 July 2022 Parasitology Research (2022) 121:2481–2493