title: Stages of infection during the tripartite interaction between Xenorhabdus nematophila, its nematode vector, and insect hosts.
abstract: Bacteria of the genus Xenorhabdus are mutually associated with entomopathogenic nematodes of the genus Steinernema and are pathogenic to a broad spectrum of insects. The nematodes act as vectors, transmitting the bacteria to insect larvae, which die within a few days of infection. We characterized the early stages of bacterial infection in the insects by constructing a constitutive green fluorescent protein (GFP)-labeled Xenorhabdus nematophila strain. We injected the GFP-labeled bacteria into insects and monitored infection. We found that the bacteria had an extracellular life cycle in the hemolymph and rapidly colonized the anterior midgut region in Spodoptera littoralis larvae. Electron microscopy showed that the bacteria occupied the extracellular matrix of connective tissues within the muscle layers of the Spodoptera midgut. We confirmed the existence of such a specific infection site in the natural route of infection by infesting Spodoptera littoralis larvae with nematodes harboring GFP-labeled Xenorhabdus. When the infective juvenile (IJ) nematodes reached the insect gut, the bacterial cells were rapidly released from the intestinal vesicle into the nematode intestine. Xenorhabdus began to escape from the anus of the nematodes when IJs were wedged in the insect intestinal wall toward the insect hemolymph. Following their release into the insect hemocoel, GFP-labeled bacteria were found only in the anterior midgut region and hemolymph of Spodoptera larvae. Comparative infection assays conducted with another insect, Locusta migratoria, also showed early bacterial colonization of connective tissues. This work shows that the extracellular matrix acts as a particular colonization site for X. nematophila within insects.
authors.affiliation: Laboratoire GPIA, Université Montpellier II-CNRS-IFREMER (UMR5171), Montpellier, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Sicard
authors.forename: Karine
authors.initials: K
authors.lastname: Brugirard-Ricaud
authors.forename: Sylvie
authors.initials: S
authors.lastname: Pagès
authors.forename: Anne
authors.initials: A
authors.lastname: Lanois
authors.forename: Noel E
authors.initials: NE
authors.lastname: Boemare
authors.forename: Michel
authors.initials: M
authors.lastname: Brehélin
authors.forename: Alain
authors.initials: A
authors.lastname: Givaudan
mesh_terms: D000818:Animals; D003238:Connective Tissue / Q000382:microbiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006458:Hemolymph / Q000382:microbiology; D007422:Intestines / Q000382:microbiology; D007814:Larva / Q000382:microbiology; D048188:Locusta migratoria / Q000382:microbiology; D008854:Microscopy, Electron; D017168:Rhabditida / Q000382:microbiology*; D018411:Spodoptera / Q000382:microbiology*; D020593:Xenorhabdus / Q000254:growth & development* / Q000472:pathogenicity*
chemical_list: D049452:Green Fluorescent Proteins