Helium atmospheric pressure plasma discharge (APPD) was previously shown to have insecticidal activity with a possible site of action on the insect nervous, neuromuscular system, or both. In the current study, methods to increase the insecticidal activity of plasma by using increased APPD temperature and the introduction of molecular oxygen were investigated for the first time. An increase in the helium plasma temperature from 37 to 50°C increased the insecticidal activity of plasma for the control of the German cockroach, Blattella germanica (L.); western flower thrips, Frankliniella occidentalis (Pergande); and citrus mealybug, Planococcus citri (Risso). This increase in activity could not be explained by the increase in air temperature alone, and it suggests that the enhanced insecticidal activity resulted from increased ionization of the APPD and ion bombardment of the insect. Emission spectroscopy showed that the introduction of 0.5% oxygen into helium plasma produced ionic molecular oxygen at 559.7 and 597.3 nm. The introduction of oxygen to the APPD greatly increased the insecticidal activity of plasma for the citrus mealybug but not the German cockroach or western flower thrips. For the mealybug as an example, the mortality of a 60-s exposure of 37°C helium plasma was 0% at 1 h after exposure and 100% under the same conditions after the introduction of oxygen. It seems that increases in temperature and the introduction of oxygen even at low levels can increase the insecticidal activity of plasma to varying degrees depending on the insect species. The symptomology of cockroach death for both hot plasma and plasma containing trace amounts of molecular oxygen continued to suggest that the site of action of APPD is the insect nervous system, neuromuscular system, or both.