Ecological immunology is based upon the notion that activation and use of the immune system is costly and should thus be traded off against other energy-demanding aspects of life history. Most of the studies on insects that have examined the possibility that mating results in trade-offs with immunity have shown that mating has immunosuppressive effects. The connection between mating and immunity has traditionally been investigated using indirect measures of immunity. However, studies that have assessed the effects of mating on the resistance against real pathogens have had conflicting results. A previous study on Tenebrio molitor showed that copulation decreases phenoloxidase activity in the haemolymph, and concluded that copulation corrupts immunity in this species. In the present study we tested whether mating also affects the ability of T. molitor to resist the entomopathogenic fungus, Beauveria bassiana. Interestingly, we found that mating enhanced resistance against the fungal infection and that the effect was stronger on males than females. Furthermore, we found that male beetles were overall more susceptible to the fungal infection than were females, indicating an immunological sex difference in immunity. Our study highlights the importance of the use of real pathogens and parasites in immuno-ecological studies.

In order to demonstrate the involvement of nitric oxide in the defence systems of marine bivalves, we investigated the production of superoxide and nitrite, following in vitro phorbol myristate acetate stimulation of Mytilus galloprovincialis, Crassostrea gigas and Ruditapes decussatus haemocytes. Whereas M. galloprovincialis and C. gigas haemocytes were found to produce superoxide and nitrite, R. decussatus haemocytes were found to be unable to generate either of these mediators. Nitrite is a stable end product of nitric oxide and peroxynitrite as well; it appeared therefore that some marine bivalves, to kill microbial pathogens, use NADPH-oxidase and nitric oxide-synthase pathways. This was confirmed at an experimental level where inhibitors of both enzymatic pathways blocked the production of nitrite. Moreover, this notion was strengthened by the inability of the haemocytes from R. decussatus, which cannot produce superoxide, to release nitrite when stimulated.