Ultrastructure and molecular characterization of the microsporidium, Nosema chrysoperlae sp. nov., from the green lacewing, Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae) used for biological pest control.

Lacewing larvae are generalist predators that are commercially available for aphid control on a variety of crops in both Europe and North America. Although lacewings are known for their symbiotic association with yeasts and bacteria, there are few reports of microsporidia in these natural enemies. An undescribed microsporidium was found in Chrysoperla carnea (Stephens) during the routine examination of specimens that were obtained from a commercial insectary for biological pest control. The objective of this study was to describe the pathogen by means of ultrastructure, molecular characterization and tissue pathology. All stages of the microsporidium were diplokaryotic and developed in direct contact with the host cell cytoplasm. Merogony and sporogony were not observed. Mature spores measured 3.49±0.10×1.52±0.05μm and had an isofilar polar filament with 8-10 coils that were frequently arranged in a single row, although double rows were also observed. Spores contained a lamellar polaroplast and a relatively small and inconspicuous polar vacuole was observed in the posterior region of about half of the spores that were examined. Tubular structures, similar in appearance to those in Nosema granulosis were observed in both sporonts and in spores. A cluster of small tubules was also observed in the posterior region of some spores. Microsporidian spores were observed in cells of the proventriculus, diverticulum and in epithelial cells of the posterior midgut. The Malpighian tubules, ileum, and rectum were heavily infected. Spores were also observed in the fat body, peripheral region of the ganglia, within and between the flight muscles, and beneath the cuticle. Although the tissues adjacent to the ovaries were heavily infected, microsporidian spores were not observed within the developing eggs. Pathogen transmission was not studied directly because it was difficult to maintain microsporidia-infected C. carnea in the laboratory. The presence of microsporidian spores in the alimentary canal suggests that the pathogen is transmitted per os and horizontal transmission may occur when infected larvae or adults are cannibalized by uninfected larvae. Molecular analysis of the microsporidian genome showed that the pathogen described in this study was 99% similar to Nosema bombycis, N. furnacalis, N. granulosis and N. spodopterae. Based on information gained during this study, we propose that the microsporidium in C. carnea be given the name Nosema chrysoperlae sp. nov.
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