As obligate blood-feeding arthropods, ticks transmit pathogens to humans and domestic animals more often than other arthropod vectors. Livestock farming plays a vital role in the rural economy of Pakistan, and tick infestation causes serious problems with it. However, research on tick species diversity and tick-borne pathogens has rarely been conducted in Pakistan. In this study, a systematic investigation of the tick species infesting livestock in different ecological regions of Pakistan was conducted to determine the microbiome and pathobiome diversity in the indigenous ticks. A total of 3,866 tick specimens were morphologically identified as 19 different tick species representing three important hard ticks, Rhipicephalus, Haemaphysalis and Hyalomma, and two soft ticks, Ornithodorus and Argas. The bacterial diversity across these tick species was assessed by bacterial 16S rRNA gene sequencing using a 454-sequencing platform on 10 of the different tick species infesting livestock. The notable genera detected include Ralstonia, Clostridium, Staphylococcus, Rickettsia, Lactococcus, Lactobacillus, Corynebacterium, Enterobacter, and Enterococcus. A survey of Spotted fever group rickettsia from 514 samples from the 13 different tick species generated rickettsial-specific amplicons in 10% (54) of total ticks tested. Only three tick species Rhipicephalus microplus, Hyalomma anatolicum, and H. dromedarii had evidence of infection with “Candidatus Rickettsia amblyommii” a result further verified using a rompB gene-specific quantitative PCR (qPCR) assay. The Hyalomma ticks also tested positive for the piroplasm, Theileria annulata, using a qPCR assay. This study provides information about tick diversity in Pakistan, and pathogenic bacteria in different tick species. Our results showed evidence for Candidatus R. amblyommii infection in Rhipicephalus microplus, H. anatolicum, and H. dromedarii ticks, which also carried T. annulata. Ticks are known for their negative impact on animal and human health through infestation and are capable of transmitting a wide range of pathogens including protozoan, viruses, and bacteria such as the spirochetes and rickettsiae. Ticks are widely distributed in different ecological, and geographical regions of Pakistan. Tick-borne diseases, such as bovine babesiosis and theileriosis have been reported in Pakistan. Crimean-Congo hemorrhagic fever (CCHF), a tick-borne viral disease in Pakistan, affects individuals who have close contact with livestock, including those working in slaughterhouses, veterinary practices, and hospitals. In Pakistan, CCHF is vectored by Hyalomma tick species, and therefore a significant threat could be posed to human health. However, there is a paucity of data on tick-borne zoonoses in Pakistan. In the current study, tick species infesting livestock across the Pakistan were catalogued, and molecular approaches were utilized to gain an insight into the microbiome associated with tick species. The results of our study revealed 19 tick species infesting livestock, and rich diversity of microbial communities associated with the ticks. Spotted Fever Group rickettsia, and Theileria annulata specific quantitative PCR assay confirmed the infection in Hyalomma tick species. To the best of our knowledge, this is the first detailed molecular study on tick species infesting livestock in Pakistan, and in depth vector competence studies should be conducted.

Ticks and tick-borne diseases (TBDs) affect the productivity of bovines in tropical and subtropical regions of the world, leading to a significant adverse impact on the livelihoods of resource-poor farming communities. Globally, four main TBDs, namely anaplasmosis, babesiosis, theileriosis, and cowdriosis (heartwater) affect bovines, and the former three are of major economic importance in bovines in Pakistan. Given that the livestock sector has become an integral part of Pakistan’s economy and a large number of dairy cattle are being imported into the country, in order to meet an increasing demand of milk and milk products, it is timely to review current status of bovine TBDs in Pakistan and to identify gaps in the knowledge of TBDs and their control. Although there has been a recent increase in the number of studies of TBDs in this country, information on their prevalence, distribution, tick vectors, and control is limited. This article provides a brief background on key bovine TBDs and ticks and reviews the current status of bovine TBDs in Pakistan to identify gaps in knowledge and understanding of these diseases, propose areas for future research and draw attention to the need for improved tools for the diagnosis and control of TBDs in this country.

A battery trial was conducted to evaluate the drug sensitivity in Eimeria tenella field isolates against the commonly used anticoccidials salinomycin