Evaluation of the Replication, Pathogenicity, and Immunogenicity of Avian Paramyxovirus (APMV) Serotypes 2, 3, 4, 5, 7, and 9 in Rhesus Macaques

Khattar, Sunil K.; Nayak, Baibaswata; Kim, Shin-Hee; Xiao, Sa; Samal, Sweety; Paldurai, Anandan; Buchholz, Ursula J.; Collins, Peter L.; Samal, Siba K.

The nasal wash, BAL, tracheal lavage and fecal samples from each monkey were clarified by centrifugation, and supernatants were collected. The clarified samples were diluted in serial 10-fold dilutions in PBS. The dilutions 10 0 , 10 1 , 10 2 , 10 3 and 10 4 (0.1 ml each) were inoculated in triplicate into the allantoic cavity of 9day-old embryonated chicken eggs. Eggs were incubated at 37uC for 4 days. Allantoic fluid was collected from each egg and the presence of virus was detected by hemagglutination (HA) test with 0.5% chicken RBC. The 50 percent egg infectious dose (EID 50 ) virus titer (expressed as log 10 EID 50 per mL) was determined by the method of Reed and Muench [48] . Replication of APMV-5 in rhesus monkeys was determined by titration in Vero cells, followed by neuramindase (NA) assay as described by Huang et al., 2004 [49] .