Psyllaephagus bliteus courtship and mating behavior were analyzed through direct observation under stereoscopic microscope ZEISS, Leica Application Suite (version 2.1.0) software. Adults parasitoids were sexed after emergence (Riek 1962) and fed with honey bee. After this, the behavior of 50 unmated couples (1 male: 1 female) of P. bliteus with different ages and 20 combinations (2 males: 1 female) were analyzed. Then at 1:1, fifty couples were formed to 10 couples of newly-emerged parasitoids, 10 of 24 h-old females and newly-emerged males, 10 of newly-emerged females and 24 h old males, 10 of 48 h old females and newly-emerged and males, and 10 of newly-emerged females and 48 h old males. At 2:1, all 20 combinations of parasitoids were newly emerged. Pairs of parasitoids were formed and analyzed daily for 15 d, without replacing the dead males or females (Table 1). The experiments were carried out at 25 ± 2°C with one pair or combination per Petri dish (5 cm diameter) for 20 min. Males and females were individualized until the moment of the test and after the analysis period. The behavioral sequences (identification, courtship, mating and postmating) associate with 474 observations in the laboratory, and successful and unsuccessful mating attempts were analyzed every day until one died, as well the period for the male and female to find each other, number of matings (duration), and postmating behaviors were recorded.

Longevity of Psyllaephagus bliteus (Hymenoptera: Encyrtidae) couples in days (mean + SE)

Preliminary assays did not indicate variations in parasitoid behavior and mating activity in the morning, afternoon, twilight, or night during observations for 60, 40, 30, or 20 min. The male parasitoid was not more active at any period, to find the females and to try to mate.

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