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The present study was planned to investigate pharmacokinetics of levofloxacin following a single dose (3 mg kg-1) intravenous (i.v.) and subcutaneous (s.c.) administration in sheep with febrile condition. Plasma samples were collected after each treatment in crossover design and analyzed for drug concentration using High Performance Liquid Chromatography (HPLC). Following intravenous administration, the drug was rapidly and widely distributed in the body with distribution and elimination half-lives of 0.24±0.03 and 2.15±0.07 h, respectively. The high volume of distribution of the drug with total clearance of 0.42±0.03 L h-1 kg-1 was observed. The drug was rapidly absorbed from subcutaneous site of administration and maximum drug concentration (Cmax) of 1.60±0.09 μg mL-1 was achieved at 1 h (Tmax). The elimination half-life and total body clearance of the drug was comparative to those observed following intravenous injection. The bioavailability (F) of the drug following subcutaneous administration in febrile sheep was 89.20±3.84%. Pharmacokinetic characteristics and absence of adverse reactions in the present study revealed that levofloxacin may be a potentially useful drug to treat bacterial diseases in sheep having acute phase reaction.