Past decades have represented great challenges for most beekeepers around the world. The number of beekeeping communities is in decline, and the global Varroa destructor infestation is one of the major causes. Beekeepers are still most commonly using synthetic acaricides to combat contagion. But the increasing presence of mite resistance, as well as growing number of studies suggesting potentially harmful effects that synthetic acaricides may have on bees and on the quality of bee products, increases the interest in alternative methods. Certain essential oils and their substances have shown great acaricidal activity, one of which is the monoterpenic phenol carvacrol that has yet to be investigated in this way. We examined the effect carvacrol has on the bee survival, on enzyme involved in transmission of nervous signal (acetylcholinesterase; AChE) and on enzyme involved in detoxification (glutathione S-transferase; GST). Both enzyme activities were measured by means of colorimetric methods. Our results suggest that 0.01% carvacrol is a sublethal dose that does not affect bee mortality, but causes a negative effect manifested by increased AChE and GST activity. 0.001% carvacrol, on the other hand, didn’t show any adverse effects. The influence that carvacrol has on the enzyme activity level is important since AChE might represent a molecular target of carvacrol, and because, besides its classical cholinergic function, it also causes effects on different roles such as learning, memory and offspring raising. Induction of GST activity implies an induction of detoxification processes considering its one the major enzymes involved in xenobiotic defense. Considering the existence of two AChE forms, the membrane-bound and water-soluble, AChE was separated by native polyacrylamide gel electrophoresis and the potential of this method for measuring the activity of both AChE forms was investigated. Staining the gels in maleate developing buffer with 1 mM of acetylthiocholine chloride promises the best results.