Fig 1.
HTNV and PUUV infect hamsters in a dose dependent manner.
Syrian hamsters were challenged with varying concentrations of either HTNV (A) or PUUV (B) through the i.m. (left) or i.n. (right) route. Between 28 and 35 days post infection hamsters were terminally bled and N-ELISA endpoint titers (log10) were used to determine infection status. The mean titer is displayed for each group, and the limit of detection (2log10) is depicted as a dashed line.
Table 1.
ID50 and ID99 for HTNV and PUUV.
Fig 2.
Kinetics of seroconversion in low dose hamster models.
Syrian hamsters were infected either 10 PFU HTNV i.m., 500 PFU HTNV i.n., 1000 PFU PUUV i.m., or 1000 PFU PUUV i.n. Hamsters were terminally bled at various points post infection and N-ELISA endpoint titers (log10) were used to determine infection status. The mean titer (A-D), and specific OD sum ± standard error of the mean (SEM) (E-F) for the N-ELISA is displayed for each group. The limit of detection (2log10) for the N-ELISA is depicted as a dashed line. The specific OD sum represents the area under the titer curve.
Fig 3.
Viral genome is detectable in the organs and sera of infected hamsters.
Syrian hamsters were infected either 10 PFU HTNV i.m., 500 PFU HTNV i.n., 1000 PFU PUUV i.m., or 1000 PFU PUUV i.n. Hamsters were euthanized at various points post infection and heart (A), lung (B), liver (C), spleen (D), kidney (E), brain (F), sera (G), and urine (H) were evaluated by RT-PCR for the presence of viral genome. The mean ± SEM is displayed at each time point. The dashed line indicates the limit of detection (1log10).
Fig 4.
Infectious virus was isolated from the organs of HTNV i.m. infected animals.
Syrian golden hamsters were infected with 10 PFU HTNV i.m. and sacrificed at various time points post infection. Heart (A), lung (B), liver (C), spleen (D), kidney (E), and brain (F) from the time range where the organ was RT-PCR positive were sampled by plaque assay for the presence of infectious virus. The mean ± SEM is displayed at each time point. The limit of detection (1.7 log10) is depicted as a dashed line.
Table 2.
Pathology scores for HTNV IM infected hamsters.
Fig 5.
Ferrets are susceptible to HTNV and PUUV infection.
Ferrets were challenged with either 2,000 PFU HTNV i.n., or 2,000 PFU PUUV i.n. (low dose challenge arrow) and did not seroconvert. The same animals were subsequently challenged with either 200,000 PFU HTNV (A,D,G,J), 94,000 PFU PUUV Beaumont (B,E,H,K), or 164,000 PFU of PUUV Seloignes (C,F,I,L) i.m. on day 0, and immunosuppressed with 30 mg/kg cyclophosphamide on day 41. The daily weight (g) for each ferret is displayed (A-C). Seroconversion of ferrets was monitored by N-ELISA specific OD sum (D-F), and neutralizing antibody production by PsVNA50 (G-I) and PRNT50 (J-L). The dashed line represents the limit of detection for the PRNT (20) and PsVNA (20) assay.
Fig 6.
PUUV infected ferrets experience weight loss, but no other clinical signs of infection.
Ferrets were infected with 94,000 PFU PUUV Beaumont i.m. and sera was collected twice weekly. Weight (A), temperature (B), N-ELISA specific OD sum (C), N-ELISA titer (log10), (D) PsVNA50 titers (E), and PRNT50 titers (F). The dotted line represents the limit of dectection for the PRNT assay (20), the PsVNA assay (20) and the N-ELISA (2log10).
Fig 7.
PUUV infected ferrets do not exhibit signs of kidney impairment.
Ferrets were infected with 94,000 PFU PUUV Beaumont i.m. Blood and urine were collected twice weekly for analysis of kidney function by assaying proteinuria (A), hematuria (B), blood urea nitrogen (C), and creatinine (D) levels. Shaded gray areas represent the normal range [48].
Fig 8.
Marmosets are susceptible to HTNV infection.
Three marmosets were infected with 1,000 PFU HTNV i.m. Sera was collected weekly to monitor seroconversion. N-ELISA titer (log10) (A) N-ELISA specific OD sum (B) PRNT50 (C) and PsVNA50 titers(D) are displayed. Serum viremia was assayed by RT-PCR (E). The limit of detection for N-ELISA titer (2log10), PRNT (20), PsVNA (20) and RT-PCR (1.0log10) is depicted by the dotted line.