The resistance status in Hyalomma anatolicum anatolicum collected from Banaskantha district, Gujarat (India) was estimated by larval packet test (LPT) with different concentrations of amitraz (125, 250, 500, 750 and 1,000 ppm) and cypermethrin (100, 200, 300, 400 and 500 ppm). The regression graphs of mean mortality of larvae ticks were plotted against values of progressively increasing concentrations of amitraz and cypermethrin for the estimation of LC 95 values and were determined as 1,529.39 and 351.84 ppm, respectively. Further a resistance of level I was determined against cypermethrin whereas, a comparatively higher resistance level (II) was recorded against amitraz. The current study appears to be the pioneer report of amitraz and cypermethrin resistance in H. a. anatolicum from the Gujarat state, India.