Theileriosis caused by Theileria lestoquardi (malignant ovine theileriosis) in sheep and Theileriaannulata (topical theileriosis) in cattle, is a tick-borne hemoprotozoal disease that causes major economic losses inanimal production caused by a tick-transmitted haemoprotozoan Theileriaspecies, infecting various domestic and wildanimals. It is a threat to the buffalo population in Pakistan due to high mortality rates and economic losses. Extensivemixed farming in Pakistan poses threats due to mixed infections and increased mortality rates. The current case reportdescribes the mixed infection and characterization of Theileria lestoquardi with Theileria annulatain a buffalo raisedin a mixed farming setup, in Pakistan. A 6.5-year-old female buffalo presented at the Outdoor Hospital exhibitingswollen sub-scapular lymph nodes, pale mucous membranes, a high fever (41.5°C), dullness, depression, shortness ofbreath, and a decrease in milk production. Investigation through microscopy, hematology, and PCR confirmed that thebuffalo was suffering from theileriosis. A Giemsa-stained thin blood smear showed piroplasms indicating theileriosis.Hematological analysis of the blood revealed lower values for hemoglobin and red blood cells (RBCs), while highercounts of lymphocytes, neutrophils, and monocytes were observed during the infection. The PCR performed on theblood sample was positive for Theileriaspp. using RLB universal primers and T. annulata-specific primers. Thesequence revealed similarity with T. lestoquardireported in Pakistan, Iran, Tunisia, and India. The buffalo was treatedwith Bupralex®and Oxtra LA®at a dosage of 1ml/20kg of body weight I/M, respectively, along with supportivetreatment. Despite these interventions, the animal died probably due to damage to cardiac muscles. This study reportsthe first case of T. annulatamixed infection with T. lestoquardiin a buffalo in Punjab, Pakistan. It can be concludedthat mixed farming exacerbates the epidemiology and spread of ovine malignant theileriosis and mortalities possiblydue to cardiac arrest in mixed infections.
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