Rhinoncomimus latipes Korotyaev (Coleoptera: Curculionidae), a biological control agent of mile-a-minute weed, Persicaria perfoliata (L.) H. Gross, has been mass reared with no infusion of new genetic material for 8–9 yr (at least 24–36 generations), while insects from the same genetic stock have been subject to field conditions in North America for that same period of time. Our main objective was to compare the laboratory population with the field population (and in 1 yr with a Chinese field population) to determine whether genetic changes had occurred, especially ones that may reduce the effectiveness of the laboratory population when released in the field. The laboratory insects laid more eggs and had reduced survival compared with field weevils in several comparisons, and had reduced responsiveness to cues that induce reproductive diapause. Exposure to older plants had the greatest effect on induction of reproductive diapause in both laboratory and field weevils, with effects of daylength and temperature less pronounced. At least a portion of the laboratory weevil population overwintered successfully. Results suggest that it is not necessary to add wild-type genetic material to the rearing colony at this time.