Infected fire salamanders died within 7 d after a short episode of anorexia, apathy, and ataxia. The pathology consistently comprised multifocal superficial erosions and deep ulcerations
the skin all over the body. Keratinocytes with eosinophilic necrosis and marginated nuclei were at the periphery of the erosions. Each of these keratinocytes contained one centrally
located thallus, the majority being segmented (colonial thalli).
Bacteria superficially colonized the ulcers. Additionally, anywhere in the skin, small foci of keratinocytes immediately below the damaged keratin layer were found. These presented similar
eosinophilic necrosis, marginated nuclei, and centrally located colonial thalli.
The intraepidermal organisms did stain with immunohistochemistry (8) (Fig. 3A). Transmission electron microscopic examination of the skin lesions confirmed the presence
of intracellular structures consistent with the colonial thalli (Fig.3B). All animals were also screened for a wide array of other infectious diseases, but no evidence for any other pathology was found: neither PCR (9) nor quantitative PCR (qPCR) (10) suggested the presence of chytrid B. dendrobatidis DNA in the skin samples. Virological examination [including PCR for the detection of herpes viruses (11), adenoviruses (12), and ranaviruses (13) and inoculation of IgH2 (iguana heart epithelial cells)and RTG (rainbow trout gill) cell cultures for general virological
investigation] was negative. Ziehl Neelsen staining, PCR for Chlamydiaceae (14), and bacterial isolation attempts did not yield any evidence of bacterial infections.
Infected fire salamanders died within 7 d after a short episode of anorexia, apathy, and ataxia. The pathology consistently comprised multifocal superficial erosions and deep ulcerations the skin all over the body. Keratinocytes with eosinophilic necrosis and marginated nuclei were at the periphery of the erosions. Each of these keratinocytes contained one centrallylocated thallus, the majority being segmented (colonial thalli).Bacteria superficially colonized the ulcers. Additionally, anywhere in the skin, small foci of keratinocytes immediately below the damaged keratin layer were found. These presented similareosinophilic necrosis, marginated nuclei, and centrally located colonial thalli. The intraepidermal organisms did stain with immunohistochemistry (8) (Fig. 3A). Transmission electron microscopic examination of the skin lesions confirmed the presenceof intracellular structures consistent with the colonial thalli (Fig.3B). All animals were also screened for a wide array of other infectious diseases, but no evidence for any other pathology was found: neither PCR (9) nor quantitative PCR (qPCR) (10) suggested the presence of chytrid B. dendrobatidis DNA in the skin samples. Virological examination [including PCR for the detection of herpes viruses (11), adenoviruses (12), and ranaviruses (13) and inoculation of IgH2 (iguana heart epithelial cells)and RTG (rainbow trout gill) cell cultures for general virologicalinvestigation] was negative. Ziehl Neelsen staining, PCR for Chlamydiaceae (14), and bacterial isolation attempts did not yield any evidence of bacterial infections.
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