Influence of Xenorhabdus (Gamma-Proteobacteria: Enterobacteriaceae) symbionts on gonad postembryonic development in Steinernema (Nematoda: Steinernematidae) nematodes

Steinernema nematodes and their Xenorhabdus partners form an obligate mutualistic association. This partnership is insecticidal to a wide range of insects. Steinernema rely on their Xenorhabdus partner to produce toxins inside the insect cadaver to liberate nutrients from the insect, as well as antimicrobials to sterilize the cadaver, thus creating a suitable environment for reproduction. In return, Steinernema vector their Xenorhabdus between insect hosts. Disruption of this partnership may affect the success of both partners. For instance, when Steinernema associates with non-cognate symbionts, their virulence and reproductive fitness are affected. In this study, we examined the effect of symbiotic (cognate and non-cognate) and non-symbiotic bacteria on maturation time, gonad postembryonic development, and sex ratio of first-generation Steinernema adults. Two Steinernema spp. were considered: S. feltiae SN and S. carpocapsae All. In vitro assays were carried out by pairing each nematode sp. with symbiotic (cognate and non-cognate) Xenorhabdus, and with non-symbiotic bacteria (Serratia proteamaculans). Additionally, for comparative purposes, we also considered adult nematodes reared in vivo in Galleria mellonella larvae to assess nematode development under natural conditions. Results from this study showed non-symbiotic Serratia proteamaculans did not support adult development of S. feltiae but it allowed development of S. carpocapsae adults. Sex ratio decreased from 2:1 to 1:1 (female: male) when S. carpocapsae adults were reared with the non-symbiotic S. proteamaculans. Cognate or non-cognate Xenorhabdus spp. and/or strains did not change the sex ratio of any of either Steinernema spp. tested. Morphometric analysis also revealed that bacterial conditions influenced adult size and gonad postembryonic development in both Steinernema species. Body size (length and width), and gonad length in both S. feltiae males and females, were significantly reduced when reared with a non-cognate Xenorhabdus species. In S. carpocapsae, males exhibited an enhanced body size (length and width) and gonad length when reared with a non-cognate X. nematophila strain. S. carpocapsae females also exhibited an enhanced gonad length when reared with a non-cognate X. nematophila strain. S. carpocapsae males and females were underdeveloped when reared with the non-symbiotic S. proteamaculans, and exhibited reduced body sizes and gonad lengths. We conclude that development of first-generation adults of both Steinernema spp. tested, in particular time to adult maturation as well as body and gonad size were directly influenced by the bacterial symbionts they were cultured with. However, response to the culture conditions was species specific.