The effects of the botanical insecticide azadirachtin were examined on the life history traits, fecundity and immune parameters of Galleria mellonella L. (Lepidoptera: Pyralidae). We determined that for the topical application of azadirachtin, the LC 50 was 16.564 ppm; at 100 ppm the adult emergence time was prolonged, however the longevity of adults remained unchanged above sublethal concentrations. The mean number of healthy eggs and the fecundity of adults decreased, whereas the number of defective eggs increased with azadirachtin treatment. At concentrations >50 ppm female G. mellonella adults laid no eggs. Azadirachtin reduced total hemocyte counts at 24 and 48 h posttreatment, however the alterations in differential hemocyte counts were only significant at 100 ppm. Laminarin-induced nodulation response and the spreading ability of hemocytes were also suppressed with azadirachtin treatment. Our results suggest that azadirachtin, as a good candidate for integrated pest control, has the capability to affect the biological parameters and cellular immunity of the model insect G. mellonella.

Venom from the pupal endoparasitoid Pimpla turionellae L. (Hymenoptera: Ichneumonidae) contains a mixture of biologically active components, which display potent paralytic, cytotoxic, and cytolytic effects toward hosts. Here, we further investigate whether parasitism or envenomation by P. turionellae alters hemocyte numbers of its host Galleria mellonella L. (Lepidoptera: Pyralidae). Total hemocyte counts declined sharply in pupae and larvae of G. mellonella exposed to P. turionellae. These same cellular responses occurred when wasp venom was artificially injected into hosts, suggesting that venom alone induces cytotoxicity in hemocytes. Analysis of the differential hemocyte counts in untreated pupae and larvae revealed that more than half of the circulating hemocytes were granular cells followed by plasmatocytes. Parasitism reduced the number of granular cells while increasing the number of plasmatocytes. This trend was most evident at 4 h postparasitism, and a similar trend was observed with the artificial injection of high (but not low) doses of venom. When isolated larval hemocytes were exposed to a LC99 dose of venom, a differential response was observed for granular cells versus plasmatocytes. Both types of cells displayed some formation of vacuoles within the cytoplasm within 15 min posttreatment. However, the degree of vacuole formation was much more extensive in granular cells at later time points than for plasmatocytes, and granular cells seemed much more susceptible to venom as evidenced by cell death.

In parasitoid species devoid of polydnaviruses and virus‐like particles, venom appears to play a major role in suppression of host immunity. Venom from the pupal endoparasitoid Pimpla turionellae L. (Hymenoptera: Ichneumonidae) has previously been shown to contain a mixture of biologically active components, which display potent paralytic, cytotoxic, and cytolytic effects toward lepidopteran and dipteran hosts. The current study was undertaken to investigate if parasitism and/or envenomation by P. turionellae affects the frequency of apoptotic and necrotic hemocytes, hemocyte viability and mitotic indices in Galleria mellonella L. (Lepidoptera: Pyralidae) pupae and larvae. Our study indicates that parasitism and experimental envenomation of G. mellonella by P. turionellae resulted in markedly different effects on the ratio of apoptotic hemocytes circulating in hemolymph depending on the host developmental stages. The ratio of early and late apoptotic hemocytes increased in G. mellonella pupae and larvae upon parasitization and at high doses of venom when compared to untreated, null and Phosphate Buffered Saline (PBS) injected controls. In contrast, an increase in necrotic hemocytes was only observed in parasitized pupae at 24 h and no difference was observed in larvae. The lowest hemocyte viability values were observed with pupae as 69.87%, 69.80%, and 72.47% at 4, 8, and 24 h post‐parasitism. The ratio of mitotic hemocytes also decreased in pupae and larvae upon parasitization and at high doses of venom. Staining of hemocytes with annexin V‐FITC revealed green fluorescent ‘halos’ along the plasma membranes of venom treated cells within 15 min following exposure to venom. By 1 h post‐venom – treatment, the majority of hemocytes displayed binding of this probe, indicative of early stage apoptosis. These same hemocytes also displayed a loss of plasma membrane integrity at the same time points as evidenced by accumulation of propidium iodide in nuclei.

The endoparasitic wasp Pimpla turionellae L. (Hymenoptera: Ichneumonidae) injects its pupal host with venom during oviposition. Venom from P. turionellae has previously been shown to contain a mixture of biologically active components, which display potent paralytic, cytotoxic and cytolytic effects towards lepidopteran and dipteran hosts. This study was undertaken to investigate if parasitism and/or envenomation by P. turionellae affects the encapsulation and melanization responses of its host Galleria mellonella L. (Lepidoptera: Pyralidae) in larval and pupal stages. Analysis of the effects of venom on encapsulation and melanization of the Sephadex A‐25 beads revealed that the number of beads strongly encapsulated and melanized were reduced by more than 50% at 4 and 24 h post‐venom injection into pupae. Injection of a lethal dose of venom (0.5 venom reservoir equivalent) in the last instar larvae was sufficient to reduce the ability of haemocytes to encapsulate the beads by more than 50% at 4 h post‐injection. Similar results were also obtained when beads were recovered from parasitized pupae indicating that parasitization by P. turionellae suppressed haemocyte‐mediated encapsulation in G. mellonella. We found that the cellular defence reactions occur more rapidly in larvae compared with pupae of G. mellonella, indicating the higher susceptibility of pupal haemocytes to parasitism and venom injection.

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