Guardians of the Herd: Molecular Surveillance of Tick Vectors Uncovers Theileriosis Perils in Large Ruminants

<i>Theileria</i> sp. (Piroplasmida: Theileriidae) is one of the most widely known infections transmitted by hard ticks (Acari: Ixodidae) and has been linked to significant economic losses across the globe. The study’s main emphasis was theileriosis, a disease that is common in Pakistan and has an incidence ranging from 0.6% to 33%. Through DNA screening of the vector ticks and host blood, this study sought to determine the risk of tick-borne theileriosis in populations of buffalos (<i>Bubalus bubalis</i>) and cattle (<i>Bos indicus</i>) in Toba Tek Singh district of Punjab, Pakistan. Identified tick species include <i>Hyalomma anatolicum</i> (35.4%), <i>Rhipicephalus (Boophilus) microplus</i> (30.2%), and <i>R. sanguineus</i> (25%). Tick specimens were collected from animals and their respective microenvironments. PCR assays targeting <i>Theileria annulata</i> were used to investigate the infection in the DNA extracted from the collected blood samples from large ruminants and salivary glands (SGs) of the <i>Hyalomma</i> ticks. The 18S rRNA of <i>T. annulata</i> was amplified using specific primers. Positive <i>T. annulata</i> amplicons were sequenced and verified using BLAST analysis. Overall, 50% of SGs contained <i>T. annulate</i> DNA. Female ticks, and those collected from cattle and from riverine environments had significantly higher (<i>p</i> < 0.05) rates of <i>Theileria</i> infection in their acini. Overall prevalence of <i>Theileria</i> infection was 35.9% in blood collected from large ruminants. Cattle had a substantially greater frequency of bovine theileriosis (43.2%) than buffalos (28.7%). Age and sex of large ruminants were significantly positively associated (<i>p</i> < 0.05) with <i>Theileria</i> infection. Furthermore, compared to non-riverine cattle (35%) and buffalo (19.5%), riverine cattle (52.2%) and buffalo (36.2%) showed a considerably higher prevalence. The results of this study, which is the first in Pakistan to examine the blood of large ruminants and vectorial function of Ixodid ticks in the transmission of <i>T. annulata</i> along with associated risk factors, offer an important insight for risk assessment of <i>Theileria</i> infection in livestock using vectorial infectivity.