Simian adenoviruses are non-enveloped, double-stranded DNA viruses of the genus Mastadenovirus in the family Adenoviridae, with icosahedral virions of 70–90 nm in diameter. Nevertheless, a continuing surveillance of pathogens in monkeys is warranted to quickly identify possible emerging zoonotic outbreaks. We detected neutralizing antibodies against SAdV RBR-7-10 in 6.8% ( n = 103) of the monkey samples but in none of the 125 human serum samples, suggesting no cross-species transmission of SAdV RBR-7-10 occurred at this study site. The SAdV RBR-7-10 virus was used in a microneutralization assay to identify virus-specific antibodies in monkey plasma and human serum samples collected from the same areas in Prachuap Khiri Khan Province. Its full hexon gene and nearly complete DNA polymerase gene were sequenced and analyzed, and the virions were imaged by transmission electron microscopy. One SAdV-B, SAdV RBR-7-10, was isolated from a long-tailed macaque fecal sample and identified by mass spectrometry. The viruses identified in these samples included Simian adenovirus (SAdV)-A, SAdV-B, SAdV-H, Human adenovirus (HAdV)-D, HAdV-G, and a bat adenovirus species. Adenoviruses were detected in 33.3% (70 out of 210) of the fecal or rectal swab samples. A few fecal samples from captive wild monkeys (a stump-tailed macaque, pig-tailed macaques, gibbons, and a leaf monkey) were also tested. Most of the monkeys tested here were long-tailed macaques, free-ranging in areas close to human territories across four provinces: Ratchaburi, Kanchanaburi, Lopburi, and Prachuap Khiri Khan. In this study, we performed a molecular survey of adenoviruses in monkeys in Thailand. The close genetic relatedness between simian and human adenoviruses, with its associated potential for the cross-species transmission of zoonotic adenoviruses from monkeys to humans and vice versa, poses important health concerns and thus warrants research. Mastadenoviruses infect mammals, including non-human primates and humans. This family is composed of non-enveloped, double-stranded DNA viruses that infect a wide range of animals. Simian adenoviruses are in the genus Mastadenovirus of the family Adenoviridae.

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