IntroductionApitoxin comes from a Latin word and means: Apisbee and toxikon -venom, being produced by bees as a way of defending and protecting the hive. The Apis mellifera apitoxin composition varies according to the subspecies development stage and habits. The primary findings are variations in the concentrations of apitoxin proteins throughout the seasons of the year, showing an influence of the environment (Abreu, 2010).The apitoxin injected into a sting contains about 50 mg of dry matter. The major proteins present are melittin; 50% of the dry weight of apitoxin (DWV), phospholipase A2 (DWV 12%), mast cell factor degranulation (3% DWV), hyaluronidase (3% DWV) and apamin (2% DWV). Moreover, AbstractThe aims of this study were to determine the period effects (morning and afternoon) and harvest time (30 and 60 minutes) in the apitoxin production as well as the management effects in the expression of a stress related gene. Five Apis mellifera L. beehives were used. The harvest of apitoxin and honeybees occurred three times a week (morning and afternoon at 09h00 and 14h00), according to the following treatments (period and apitoxin harvest time): T1: morning/30 minutes, T2: morning/60 minutes; T3: afternoon/30 minutes; T4: afternoon/60 minutes. The apitoxin was collected by electric collectors. The stress level was monitored by defensin gene expression, using actin gene as control. The results were evaluated by ANOVA followed by Tukey-Kramer test (p ≤ 0.05). It can be concluded that the better period and time to apitoxin harvest is in the morning for 60 minutes, associated to minor stress for honeybees.