Fig 1.
Confocal microscopy analysis of embryos of Gallus gallus domesticus 48 hpi with Yellow Fever 17DD virus.
Mesenchymal cells in leg skeletal muscle (A) and in heart (B). Yellow fever virus in green, nuclei stained with DAPI in blue.
Fig 2.
Confocal microscopy analysis of Gallus gallus domesticus 72 hpi with Yellow Fever 17DD virus.
Infected skeletal muscle bundles (A). The viral protein form clusters in the cytoplasm and follow the muscular striations; Infected muscular fibers and mesenchymal cells adhering to infected and uninfected muscular fibers (B); Infected muscular cells in the heart (C). Infected kidney tubular epithelium cells (D). Yellow fever virus proteins in green, nuclei stained with DAPI in blue and desmin in red.
Fig 3.
Confocal microscopy analysis of brain in Gallus gallus domesticus 72 hpi with Yellow Fever 17DD virus.
Infected nervous tissue cells in the brain (A); Details of infected neurons in the brain (B). Yellow fever virus proteins in green, nuclei stained with DAPI in blue.
Fig 4.
Confocal microscopy analysis of different tissues in Gallus gallus domesticus 72 hpi with Yellow Fever 17DD virus.
Infected cells in dermal connective tissue (A); Infected lung parenchyma cells (B); Infected cells in the muscular layer of the gizzard (C); Infected fibroblastoid cells in the yolk stalk (D). Yellow fever virus proteins in green, nuclei stained with DAPI in blue.
Fig 5.
Brightfield microscopy analysis of Gallus gallus domesticus 72hpi with Yellow Fever 17DD virus.
Apoptotic corpuscles in muscle bundles (A), and in the kidney tubular epithelium (B). Apoptotic nuclei are indicated by black arrow (→). Hematoxylin and eosin stain.
Fig 6.
Detection of YF 17DD viral RNA by nested-PCR analysis.
The RNA was extracted from formalin-fixed paraffin-embedded tissues. YF 17DD amplicons were assayed by means of agarose gel electrophoresis. The above figure show samples from a chicken embryo infected for 72 hours, and the bottom figure 96 hours. The lanes correspond to the following specimens: (1) empty; (2) and (3) head; (4) and (5) legs; (6) and (7) wings; lanes from (8) to (15) trunks; (16) and (17) vitelline membrane; (18) and (19) chorioallantoic membrane; from (20) to (23) negative control (water-inoculated animals). Even-numbered lanes indicate samples submitted to amplification of genomic RNA whereas odd-numbered lanes indicate samples submitted to amplification of the replicative intermediate RNA. The molecular length markers are indicated on the left of the figure. The black arrow indicates the 156bp amplicon obtained from the amplification of YF 17DD RNA.
Fig 7.
Brightfield microscopy analysis of Gallus gallus domesticus 96hpi with Yellow Fever 17DD virus.
Apoptotic bodies in (A) muscular bundles; (B) heart cells; (C) kidney tubular epithelium; (D) gizzard muscle; (E) lung parenchyma; and (F) brain cells. Apoptotic nuclei are indicated by black arrow (→). Hematoxylin and eosin stain.
Fig 8.
Confocal microscopy analysis of muscular tissue in Gallus gallus domesticus 96hpi with Yellow Fever 17DD virus.
Infected skeletal muscle bundles, shown viral protein aggregation (A); detail of infected muscular fibers shown perinuclear immunostaining of viral protein and clusters follow the muscular striations, longitudinal section (B) and transversal section (C); infected cardiomyocytes in the heart (D). Apoptotic bodies closer to infected cells (→).Yellow fever virus proteins in green, nuclei stained with DAPI in blue and desmin in red.
Fig 9.
Confocal microscopy analysis of brain in Gallus gallus domesticus 96 hpi with Yellow Fever 17DD virus.
Infected nervous tissue cells in the brain (A, B); Detail of infected cells in the brain congested with virus proteins (C,D). Yellow fever virus proteins in green, nuclei stained with DAPI in blue.
Fig 10.
Confocal microscopy analysis of nervous system in Gallus gallus domesticus 96 hpi with Yellow Fever 17DD virus.
Infected neurons in the spinal cord (A); infected cells in nerve bundles (B); infected cells in the dorsal root ganglion (C); infected fibroblastoid cells in the meninges (D). Yellow fever virus proteins in green, nuclei stained with DAPI in blue.