First evidence of a new simian adenovirus clustering with Human mastadenovirus F viruses.

BACKGROUND: Adenoviruses play an important role as human pathogens, though most infections are believed to be asymptomatic. The over 100 human adenovirus types are classified into seven species (A-G), some of which include simian adenoviruses. Recent findings have highlighted that simian adenoviruses have a zoonotic potential and that some human adenoviruses are likely the result of relatively recent spillover events. METHODS: In order to evaluate the risks associated with primates hunted and sold as bushmeat, multiple samples from 24 freshly killed monkeys were collected in the Republic of the Congo and tested for adenovirus DNA by PCRs targeting the conserved DNA polymerase and hexon genes. RESULTS: The DNA of a novel simian adenovirus was detected in a moustached monkey (Cercopithecus cephus) by the DNA polymerase PCR, but not by the hexon PCR. The 275 nucleotide amplicon was most closely related to members of the Human mastadenovirus F species (93% HAdV-40 and 89% HAdV-41 amino acid identity), rather than to other known simian adenoviruses. CONCLUSIONS: The phylogenetic clustering with Human mastadenovirus F sequences suggests a common ancestor, more recent than the last common ancestor of humans and moustached monkeys. The findings increase concerns about the zoonotic potential of simian adenoviruses and highlight the need for more research and surveillance on the issue.

Taxonomy proposal for Old World monkey adenoviruses: characterisation of several non-human, non-ape primate adenovirus lineages.

A species classification regarding Old World monkey adenoviruses is proposed. We determined the nucleotide sequences of PCR-amplified fragments from the genes of the IVa2, DNA-dependent DNA polymerase, penton base, and hexon proteins from every simian adenovirus (SAdV) serotype that originated from Old World monkeys for which the full genome sequence had not yet been published. We confirmed that the majority of Old Word monkey SAdVs belong to two previously established species. Interestingly, one is the most recently established human AdV species, Human mastadenovirus G, which includes a single human virus, HAdV-52, as well as SAdV-1, -2, -7, -11, -12, and -15. The other approved species, Simian mastadenovirus A includes SAdV-3, -4, -6, -9, -10, -14, and -48. Several SAdVs (SAdV-5, -8, -49, -50) together with baboon AdV-1 and rhesus monkey AdV strains A1139, A1163, A1173, A1258, A1285, A1296, A1312, A1327 and A1335 have been proposed to be classified into an additional species, Simian mastadenovirus B. Another proposed species, Simian mastadenovirus C has been described for SAdV-19, baboon AdV-2/4 and -3. Our study revealed the existence of four additional AdV lineages. The corresponding new candidate species are Simian mastadenovirus D (for SAdV-13), Simian mastadenovirus E (for SAdV-16), Simian mastadenovirus F (for SAdV-17 and -18), and Simian mastadenovirus G (for SAdV-20). Several biological and genomic properties, such as the host origin, haemagglutination profile, number of fibre genes, and G+C content of the genome, strongly support this classification. Three SAdV strains originating from the American Type Culture Collection turned out to be mixtures of at least two virus types, either of the same species (SAdV-12 and -15 types from Human mastadenovirus G) or of two different species (SAdV-5 types from Simian mastadenovirus B and Human mastadenovirus G).

A novel adenovirus species associated with an acute respiratory outbreak in a baboon colony and evidence of coincident human infection.

Adenoviruses (AdVs) are DNA viruses that infect many vertebrate hosts, including humans and nonhuman primates. Here we identify a novel AdV species, provisionally named "simian adenovirus C (SAdV-C)," associated with a 1997 outbreak of acute respiratory illness in captive baboons (4 of 9) at a primate research facility in Texas. None of the six AdVs recovered from baboons (BaAdVs) during the outbreak, including the two baboons who died from pneumonia, were typeable. Since clinical samples from the two fatal cases were not available, whole-genome sequencing of nasal isolates from one sick baboon and three asymptomatic baboons during the outbreak was performed. Three AdVs were members of species SAdV-C (BaAdV-2 and BaAdV-4 were genetically identical, and BaAdV-3), while one (BaAdV-1) was a member of the recently described SAdV-B species. BaAdV-3 was the only AdV among the 4 isolated from a sick baboon, and thus was deemed to be the cause of the outbreak. Significant divergence (<58% amino acid identity) was found in one of the fiber proteins of BaAdV-3 relative to BaAdV-2 and -4, suggesting that BaAdV-3 may be a rare SAdV-C recombinant. Neutralizing antibodies to the other 3 AdVs, but not BaAdV-3, were detected in healthy baboons from 1996 to 2003 and staff personnel from 1997. These results implicate a novel adenovirus species (SAdV-C) in an acute respiratory outbreak in a baboon colony and underscore the potential for cross-species transmission of AdVs between humans and nonhuman primates. IMPORTANCE Adenoviruses (AdVs) are DNA viruses that infect many animals, including humans and monkeys. In 1997, an outbreak of acute respiratory illness from AdVs occurred in a baboon colony in Texas. Here we use whole-genome sequencing and antibody testing to investigate new AdVs in baboons (BaAdVs) during the outbreak, one of which, BaAdV-3, came from a sick animal. By sequence analysis, BaAdV-3 may be a recombinant strain that arose from a related BaAdV found in baboons nearby in the colony (who were not sick) and yet another unknown AdV. We also found antibodies to these new BaAdVs in baboons and staff personnel at the facility. Taken together, our findings of a new AdV species as the cause of an acute respiratory outbreak in a baboon colony underscore the ongoing threat from emerging viruses that may carry the potential for cross-species transmission between monkeys and humans.

Adenoviruses in fecal samples from asymptomatic rhesus macaques, United States.

Adenoviruses can cause infectious diarrheal disease or respiratory infections in humans; 2 recent reports have indicated probable human infection with simian adenoviruses (SAdVs). To assess the possibility of animal-to-human transmission of SAdVs, we tested fecal samples from asymptomatic rhesus macaques housed in 5 primate facilities in the United States and cultured 23 SAdV isolates. Of these, 9 were purified and completely sequenced; 3 SAdV samples from the American Type Culture Collection (SAdV-6, SAdV-18, and SAdV-20) were also completely sequenced. The sequence of SAdV-18 was closely related to that of human adenovirus F across the whole genome, and the new isolates were found to harbor 2 fiber genes similar to those of human adenovirus (HAdV) strains HAdV-40 and HAdV-41, which can cause infectious diarrhea. The high prevalence of adenoviruses in fecal samples from asymptomatic rhesus macaques and the similarity of the isolates to human strains indicates the possibility of animal-to-human transmission of SAdVs.

Isolation of a novel monkey adenovirus reveals a new phylogenetic clade in the evolutionary history of simian adenoviruses.

Adenoviruses of primates include human (HAdV) and simian (SAdV) isolates classified into 8 species (Human Adenovirus A to G, and Simian Adenovirus A). In this study, a novel adenovirus was isolated from a colony of cynomolgus macaques (Macaca fascicularis) and subcultured in VERO cells. Its complete genome was purified and a region encompassing the hexon gene, the protease gene, the DNA binding protein (DBP) and the 100 kDa protein was amplified by PCR and sequenced by primer walking. Sequence analysis of these four genes showed that the new isolate had 80% identity to other primate adenoviruses and lacked recombination events. The study of the evolutionary relationships of this new monkey AdV based on the combined sequences of the four genes supported a close relationship to SAdV-3 and SAdV-6, lineages isolated from Rhesus monkeys. The clade formed by these three types is separated from the remaining clades and establishes a novel branch that is related to species HAdV-A, F and G. However, the genetic distance corresponding to the newly isolated monkey AdV considerably differs from these as to belong to a new, not yet established species. Results presented here widen our knowledge on SAdV and represents an important contribution to the understanding of the evolutionary history of primate adenoviruses.

Identification of adenoviruses in fecal specimens from wild chimpanzees (Pan trogylodytes schweinfurthii) in western Tanzania.

DNA of two distinctive adenoviruses was detected in wild chimpanzees in western Tanzania that showed clinical signs of acute, upper respiratory disease, notably coughing. The amplified sequences from part of the capsid hexon gene suggests that one virus is a novel adenovirus serotype candidate and the other virus is a species C adenovirus most closely related to recent isolates from captive chimpanzees in the United States, Simian AdV 37 with 86% nucleic acid identity and Simian AdV 40 with 95% nucleic acid identity, respectively. The species C adenovirus sequences suggest possible recombination with a human adenovirus. The source of these viruses and disease association is not known.

Molecular detection of novel adenoviruses in fecal specimens of captive monkeys with diarrhea in China.

Adenovirus (AdV) has been recently detected among monkeys with diarrhea in a major research primate colony in China. To better assess disease burden and epidemiology of adenoviruses in the colony, we examined the prevalence of this virus in fecal specimens by PCR using broadly reactive hexon gene-specific primers. Of the 29 strains that were characterized by sequence and phylogenetic analysis, we identified a broad spectrum of simian AdV (SAdV) types, including species SAdV-A (n=14) and HAdV-G (n=9). Six additional strains represented two genetic clusters distantly related to other known SAdVs. A better understanding of the epidemiology of SAdVs and their potential role in gastroenteritis is critical to the implementation of advanced prevention strategies against AdV infection in captive primates.

Isolation of enteroviruses and adenoviruses in continuous simian cell lines.

The ability of five simian cell lines--BGM, LLC-MK2, MA104, Vero and BSC-1--to isolate enteroviruses and adenoviruses from faeces was assessed and compared with that of primary or secondary rhesus monkey kidney (RhMK) and HEp2 cultures. For enteroviruses, LLC-MK2 and BGM were best of the five lines, detecting 77% and 73% respectively of 88 strains isolated, but neither was as good as RhMK, which detected 88%. Both Vero and BSC-1 detected less than 50%. All cell types were good at isolating poliovirus and coxsackie B strains, although LLC-MK2 and BGM were slightly better than MA104, Vero, BSC-1 and HEp2 for the latter. Greatest variation in sensitivity occurred with coxsackie A9 and echovirus isolates, with all lines being less sensitive than RhMK. However, the combined isolation rate for BGM, LLC-MK2 and HEp2 was the same as that for RhMK and HEp2, although the strains not detected by each cell combination were different. Both Vero and BSC-1 were reasonable for detecting adenoviruses, but neither was as good as HEp2. The reasons for variations in results, and the future use for such cell lines for isolating viruses from clinical and environmental samples, are discussed.

Maternal-fetal transmission of SIV in macaques: disseminated adenovirus infection in an offspring with congenital SIV infection.

To develop a nonhuman primate model for maternal-fetal transmission of HIV infection, we have inoculated pregnant Macaca nemestrina with uncloned SIVMne. Three animals inoculated during the third trimester delivered healthy infants. One of the three infants, a male born 31 days after the mother was inoculated with SIV, became virus-positive but failed to produce SIV-specific antibody and died with overt simian immunodeficiency and disseminated adenovirus (SV20) infection at age six and one-half months. SIV and adenovirus antigen could be demonstrated by immunohistochemical methods in multiple organ systems.

Aspiration pneumonia with adenovirus infection in a Japanese macaque (Macaca fuscata fuscata).

Pneumonia with aspiration of foreign bodies and adenovirus infection was identified in a wild Japanese macaque which died 1 month after capture. The adenovirus infection was considered to be secondary in importance to the pneumonia.

Influence of water quality on enteric virus concentration by microporous filter methods.

Four enteric viruses, poliovirus type 1, echovirus type 1, reovirus type 3, and simian adenovirus SV-11, were concentrated from seeded 1.3-liter volumes of raw, finished, and granular activated carbon-treated waters by adsorption to 47-mm-diameter (17 cm2), electropositive ( Virosorb 1MDS ) filters at pH 7.5 or electronegative ( Filterite ) filters at pH 3.5 with and without 5 mM added MgCl2, followed by elution with 0.3% beef extract in 50 mM glycine at pH 9.5. Removal of particulates from raw and finished waters by 0.2-micron prefiltration before virus addition and pH adjustment had little effect on virus concentration efficiencies. Soluble organic compounds reduced virus adsorption efficiencies from both raw and finished waters compared with granular activated carbon-treated water, but the extent of interference varied with virus type and adsorption conditions. For electropositive 1MDS filters, organic interference was similar with all virus types. For Filterite filters, organic interference was evident with poliovirus and echovirus, but could be overcome by adding MgCl2. Reovirus and SV-11 were not adversely affected by organics during adsorption to Filterite filters. Elution of reovirus and adenovirus was inefficient compared with that of poliovirus and echovirus. None of the three adsorption schemes ( 1MDS at pH 7.5 and Filterite with and without 5 mM MgCl2 at pH 3.5) could be judged superior for all viruses and water types tested.

[Identification of a new antigenic variant of simian adenovirus (SV30N)]. / Identifikatsiia novogo antigennogo varianta adenovirusa obez'ian (SV30N).

A new variant of simian adenovirus SV30-N is described. The variant is antigenically close to SV30 virus in the neutralization test but has some antigenic relationship to SA7 virus. The properties of SV30-N virus were retained after 5 clonings by the plaque method, after passage and cloning of the virus treated with immune sera to SV30 or SA7. The heteroduplex analysis showed DNA of the virus under study to be by 80% and 40% homologous to SV30 DNA and SA7 DNA, respectively, whereas no heteroduplex molecules between SV30 and SA7 DNA were found. By the set of polypeptides the antigenic variant SV30-N is close to SV30 but differs from SV30 and, particularly from SA7. Unlike SV30 and SA7 viruses, the SV30-N virus showed no oncogenic activity for hamsters.

Virus detection in monkeys with diarrhea: the association of adenoviruses with diarrhea and the possible role of rotaviruses.

To explore the role of viruses in the etiology of diarrhea in colony-reared monkeys, direct electron microscopy, the fluorescent virus precipitin test and cell culture inoculation were used to examine the stools of monkeys with and without diarrhea. The animals were predominantly rhesus with a few macaques of other species, and included infants, juveniles and adults. Adenoviruses were isolated from a higher proportion of specimens from rhesus monkeys with diarrhea (73% of specimens from infants and 78% of specimens from juveniles and adults) than from control monkeys without diarrhea (22% of specimens from infants and 26% of specimens from juveniles and adults). SV 20 was the most frequently isolated simian adenovirus type; SV 17 and SV 32 also were recovered. Noncultivable adenoviruses detectable only by electron microscopy were not seen. Although adenovirus excretion was associated with diarrhea, the causal role of adenoviruses was difficult to assess. When serial specimens from animals with chronic or intermittent episodes of diarrhea were examined, sequential infections with different viruses were found to be common. Rotaviruses were detected by electron microscopy and isolated in cell cultures from two infant rhesus monkeys with diarrhea. However, the low detection rate, together with negative serologic data on 40% of infant monkeys with diarrhea, suggested that rotaviruses were not the major cause of gastroenteritis in the monkeys under study.

Three serologic types of adenovirus infections of owl monkeys.

From a colony of 131 owl monkeys, adenovirus was isolated 68 times from 433 oropharyngeal and rectal swab sample pairs collected over a 2-year period. A total of 63 of 68 adenovirus isolates were grouped by neutralization into 1 of 3 types designated owl monkey adenovirus types (OMAV Ty) I, II, and III. The OMAV Ty I and Ty II were identified by neutralization with antiserum to squirrel monkey adenovirus type I and adenovirus SV-11, respectively. The OMAV Ty III was partially neutralized by antiserum to OMAV Ty II. The OMAV Ty I was isolated from 8 of 30 newly imported owl monkeys within 3 weeks of arrival. A total of 13 of 26 owl monkeys had a fourfold or greater rise in serum-neutralization (SN) titer with 25 of 27 developing SN titers of greater than or equal to 1:80. A total of 40 of 87 other owl monkeys in the colony had SN titers of greater than or equal to 1:5. The OMAV Ty II was isolated from 51 (12%) of 433 swab samples pairs. Virus was isolated from 26 owl monkeys with 13 having persistent infections. Viral isolations spanned a 10- to 17-month period for five of these owl monkeys. In 114 owl monkeys, 101 (86%) had SN titers to OMAV Ty II greater than or equal to 1:20. The OMAV Ty III was isolated from three owl monkeys. Of 114 owl monkeys, 93 (82%) had SN titers to OMAV Ty III of greater than or equal to 1:20. Although the distribution of SN titers were similar for OMAV Ty II and Ty III, 42 (37%) of 114 owl monkeys had eightfold or greater differences in SN titers between OMAV Ty II and Ty III.

Improved methods for detecting enteric viruses in oysters.

New and improved methods for concentrating enteroviruses, reoviruses, and adenoviruses from oysters have been developed and evaluated. Viruses are efficiently adsorbed to homogenized oyster meat by adjusting the homogenate to pH 5.0 and a conductivity of less than or equal to 2,000 mg of NaCl per liter. After low-speed centrifugation, the virus-free supernatant is discarded and the viruses are eluted from the sedimented oyster solids with pH 7.5 glycine-NaCl having a conductivity of 8,000 mg of NaCl per liter. The oyster solids are removed by low-speed centrifugation and filtration, and the viruses in the filtered supernatant are concentrated to a small volume by either ultrafiltration or acid precipitation at pH 4.5. The concentrate is treated with antibiotics and inoculated into cell cultures for virus isolation and quantitation. When these methods were tested with oysters experimentally contaminated with polioviruses, reoviruses, and adenoviruses, recovery efficiencies averaged about 46%. With the exception of virus assay and quantitation, these methods are simple and inexpensive enough to be done in typical shellfish microbiology laboratories.

Persistent shedding of adenovirus in urine of chimpanzees.

A new adenovirus, designated Pan 11, was isolated repeatedly from the urine of several chimpanzees for more than 1 year. One chimpanzee had chronic interstitial nephritis; the others were healthy. Most chimpanzees tested had neutralizing antibodies to Pan 11 virus in the serum; three people who worked with chimpanzees also acquired antibodies. Transplantable rhabdomyosarcomas developed in hamsters inoculated as newborns with Pan 11 virus.

Necrotizing pancreatitis due to simian adenovirus type 31 in a rhesus monkey.

Simian adenovirus type 31 was isolated from pancreatic tissue of an 8-year-old male rhesus monkey that died as the result of acute necrotizing pancreatitis. Histologically, the pancreas showed wide-spread necrosis, extensive infiltration of polymorphonuclear leukocytes, and nuclear inclusions in pancreatic acinar cells. Large numbers of adenovirus particles were demonstrable in the acinar cells by electron microscopy. This is the second case of adenovirus-associated pancreatitis we have found in rhesus monkeys and the first case in which we attempted to isolate the virus. Our data indicate that adenovirus pancreatitis is a distinct entity in rhesus monkeys and may occur with greater frequency than is generally believed.