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Fig 1.

Experimental design.

Guinea pig infection and tick blood meal.

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Fig 2.

Experimental design: moulting and testing samples.

Real-time PCR and culture: □; Transmission Electron Microscopy: ●; Fluorescence In Situ Hybridization: ▲.

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Table 1.

Engorged Dermacentor reticulatus female ticks after feeding on guinea pigs infected with F. tularensis subsp. holarctica.

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Table 2.

Engorged Ixodes ricinus female ticks after feeding on guinea pigs infected with F. tularensis subsp. holarctica.

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Fig 3.

TEM micrographs of Ixodes ricinus pre-vitellogenic oocyte after feeding on guinea pigs infected with F. tularensis subsp. holarctica.

The morphology of some inclusions is typical of a Gram-negative bacterium, and their size is congruent with that of bacteria from the Francisella genus. m: mithocondrium, b: bacterium. Scale bar: A 0.22 μm, B 1.1 μm.

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Fig 4.

TEM micrographs of Ixodes ricinus pre-vitellogenic oocyte after feeding on guinea pigs infected with F. tularensis subsp. holarctica.

Note the presence of residual bodies into the large, phagocytic vacuole. Oo: oocyte, mv: microvilli, Tp: tunica propria. Scale bar: 0.37 μm.

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Fig 5.

FISH staining with a probe specific for Francisella tularensis in Dermacentor reticulatus oocytes after feeding on guinea pigs infected with F. tularensis subsp. holarctica.

(A) Image obtained from light transmission; (B) Overlay image with blue signal for F. tularensis (23S rRNA probe for F. tularensis labelled with the fluorochrome Cy5) and red signal for universal eubacterial probe EUB338; (C) 23S rRNA probe for all F. tularensis; (D) universal eubacterial probe EUB338. Scale bar: 200 μm.

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Fig 6.

FISH staining with a probe specific for Francisella tularensis in Ixodes ricinus salivary glands from female after feeding on guinea pigs infected with F. tularensis subsp. holarctica.

(A) Image obtained from light transmission; (B) Overlay image with blue signal for F. tularensis (23S rRNA probe for F. tularensis labelled with the fluorochrome Cy5) and red signal for universal eubacterial probe EUB338; (C) 23S rRNA probe for F. tularensis; (D) universal eubacterial probe EUB338. Scale bar: 100 μm.

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Fig 6 Expand