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. 2014 Feb 10;5:43. doi: 10.3389/fmicb.2014.00043

Figure 3.

Figure 3

N. marisprofundi is associated with D. marci reproductive organs. (A) A diagram of infected D. marci male and female. Gonads, male vas deferens, and female uterus are labeled in blue. Microsporidia spores are labeled in red. The bar graphs show the tissue-specific patterns of spore distribution in males and females, numbers are not exclusive. (B) Nomarski DIC of a moderately infected female. E, eggs; V, vulva; arrow, microsporidia spores. (C) N. marisprofundi infection of male reproductive organs. Microsporidia spores are distributed at the end of the vas deferens (arrow) and at the dorsal posterior side; S, male spicule; magnification as in B. (D) N. marisprofundi spores dissected from a female genital tract and stained by 4′,6-diamidino-2-phenylindole (DAPI). (E) DAPI staining (blue) revealed nucleus-like structure inside the spores (arrowhead) magnification as in (D). (F) Calcofluor white staining (blue) of N. marisprofundi spores (arrows) attached to a dissected worm cuticle. (G) In situ hybridization with N. marisprofundi SSU rRNA probe (red) and DAPI staining (blue) of an infected female fixed by paraformaldehyde to detect specifically the vegetative stages (see Materials and Methods section in Supporting Information). Vegetative stages (arrowheads) are positioned in the vicinity of spores. E developing embryo. Scale bars are 20 μm (B,F,G); 5 μ m (D); insert 2.5 μm (G).