Tensiomyography (TMG) has been described as an effective method for identifying differences in muscle response to athletic training stimuli. However, there is a lack of studies that evaluated changes in muscle contractile properties in response to anaerobic effort and related these changes to Wingate test parameters in more than one lower limb superficial muscle. The main contribution of this work is to evaluate changes in the contractile properties of muscles in response to anaerobic exercise. In a group of 20 physically active men, the body composition analysis was performed using a Tanita DC-360 device. The anaerobic effort was assessed by a Wingate test (30 s), and the contractile properties of muscles were measured using a TMG S2 device. The TMG parameters were measured in selected superficial muscles of the dominant lower limb. The study also calculated indices of the differences (d) and the size of the effect (r). An anaerobic effort in the form of the Wingate test resulted in greater stiffness (Dm) in the vastus lateralis muscle (p = 0.0365; r = 0.47) and a delayed response to stimulus (Td) in the vastus lateralis (p = 0.0239; r = 0.51) and vastus medialis (p = 0.0031; r = 0.66) muscles. The half relaxation time (Tr) (p = 0.0478; r = 0.44) and sustained contraction time (Ts) (p = 0.0276; r = 0.49) in the rectus femoris muscle were also increased. In contrast, a prolonged stimulus-response time (Td) with a decrease in sustained contraction time (Ts) was observed in the gastrocnemius lateralis (Td: p = 0.0054; r = 0.64 and Ts: p = 0.0012; r = 0.74) and gastrocnemius medialis (Td: p = 0.0229; r = 0.52 and Ts: p = 0.0054; r = 0.64) muscles. A significant decrease in contraction time (Tc) (p = 0.0051; r = 0.63) occurred only in the soleus muscle. In addition, significant correlations were shown between selected changes in contractile properties of muscle and parameters of the Wingate anaerobic test. Anaerobic exercise has a significant effect on changes in skeletal muscle contractility parameters. TMG is an effective method for identifying differences in muscle response to sports training stimuli.