Figure 1.
Clinical and epidemiologic features of the titi monkey outbreak.
(A) Map of the titi monkey cages situated in one quadrant of a building, showing the locations of asymptomatic, at-risk monkeys (brown or green), affected surviving monkeys (black), and monkeys who died from their illness (skeleton). 3 monkeys were moved into the building (arrows pointing down and to the left) and 4 monkeys out of the building (arrows pointing up and to the right) during the 3rd week of the outbreak. The upper left photograph shows an image of an adult male titi monkey and his infant. The upper right inset shows the location of the titi monkey cages relative to other rhesus monkey cages in the building. Asymptomatic monkeys with positive serum antibody titers to TMAdV 4 months after the outbreak are shown in green. (B) Epidemic curve of the outbreak, with the number of cases in blue and cumulative attack rate in red. (C) Anteroposterior chest radiograph of an affected titi monkey, showing bilateral basilar infiltrates and a prominent right middle lobe consolidation. (D) 1 – gross photograph of lungs at necropsy; the lungs failed to fully collapse upon opening the chest, and a single ∼1.5 cm focus of dark red discoloration (hemorrhage) can be seen in the left caudal lobe. 2 – photomicrograph of H&E stained lung tissue showing a severe diffuse necrotizing bronchopneumonia characterized by the presence of hemorrhage and intranuclear inclusions (arrows). 3 – photomicrograph of H&E stained liver tissue showing a multifocal necrotizing hepatitis with numerous intranuclear inclusions (arrows). 4 – transmission electron micrograph of an affected lung alveolus (scale bar = 1 µm) filled with adenovirus-like particles (inset, scale bar = 0.1 µm).
Figure 2.
Discovery and whole-genome characterization of the novel adenovirus TMAdV.
(A) The locations of the 4 Virochip probes derived from adenovirus sequences and used to detect TMAdV are mapped onto the ∼37 kB genome. The 4 Virochip probe sequences are also aligned with the corresponding sequence in the TMAdV genome, with mismatches highlighted in pink. (B) Coverage map of deep sequencing reads corresponding to TMAdV using BLASTN (blue) and TBLASTX (transparent blue) alignments to SAdV-18. The actual coverage achieved by deep sequencing as determined by alignments to the fully sequenced genome of TMAdV is much higher (red). (C) Genome organization of TMAdV. Predicted protein coding regions are shown as boxes. Boxes above the central black line represent open reading frames (ORFs) that are encoded on the forward strand, while boxes underneath the black line represent reverse-strand encoded ORFs. Early region ORFs are shaded in gray. The x-axis refers to the nucleotide position along the ∼37k genome of TMAdV. Abbreviations: FAdV, fowl adenovirus; SAdV, simian adenovirus; PAdV, porcine adenovirus; HAdV, human adenovirus, TMAdV, titi monkey adenovirus.
Table 1.
PCR screening for TMAdV.
Figure 3.
Whole-genome phylogenetic analysis of TMAdV.
The whole-genome nucleotide phylogenetic tree is reconstructed from a multiple sequence alignment of all 95 unique, fully-sequenced adenovirus genomes in GenBank and TMAdV. Both rectangular cladogram and radial tree layouts are displayed. The branch corresponding to TMAdV is highlighted in boldface red. Abbreviations: HAdV, human adenovirus; SAdV, simian adenovirus; MAdV, mouse adenovirus, FrAdV, frog adenovirus; TAdV, turkey adenovirus; SnAdV, snake adenovirus; DAdV, duck adenovirus; OAdV, ovine adenovirus; BAdV, bovine adenovirus; PAdV, porcine adenovirus; TSAdV, tree shrew adenovirus; CAdV, canine adenovirus.
Figure 4.
Scanning pairwise alignment of representative adenoviruses with TMAdV.
The scanning nucleotide pairwise identities of TMAdV relative to representative human (yellow) or simian (brown) adenoviruses in species A–G, porcine adenovirus (red), and fowl adenovirus (green) are shown. The window size is 400 bp with a step size of 40 bp. The x-axis refers to the nucleotide position along the ∼37 k genome of TMAdV. Abbreviations: HAdV, human adenovirus; SAdV, simian adenovirus; PAdV, porcine adenovirus; FAdV, fowl adenovirus.
Table 2.
Amino acid identity of TMAdV relative to other adenoviruses.
Figure 5.
Growth and propagation of TMAdV in cell culture.
The flow chart displays 10 passages (P1–P10) of TMAdV cultured in human lung adenocarcinoma (A549, orange), primary rhesus macaque kidney (PMK, brown), or established African green monkey kidney (BSC-1, green) cells.
Figure 6.
Seroprevalence of TMAdV in humans and monkeys.
Sera from titi monkeys (circles), rhesus macaques (squares), and humans (triangles) were tested for antibodies to TMAdV by virus neutralization. Arrows designate pre-outbreak and post-outbreak serum samples from the same individual monkey. Pre-outbreak serum samples were previously banked in 2007. Sera from CNPRC personnel and close contacts (orange triangles) were collected 4 months post-outbreak, except for the two family members of the clinically ill researcher, whose sera were collected 1 year post-outbreak. *, clinically ill researcher; **, family member of the researcher, who was also sick. Abbreviations: CNPRC, California National Primate Research Center; NEG, negative.