The Prevalence of Staphylococcus aureus and the Occurrence of MRSA CC398 in Monkey Feces in a Zoo Park in Eastern China

SIMPLE SUMMARY: The increasing observation of methicillin-resistant Staphylococcus aureus (MRSA) in wildlife species has raised the concern of its impact on the animal health and the potential of zoonotic transmission. The molecular types of MRSA and S. aureus in wild animals has been found to mix with several molecular types from humans and livestock, indicating a dynamics of S. aureus transmission between these animals and humans. Thus, it is important to investigate the epidemiological relationship of MRSA between zoo animals and humans. To investigate the prevalence and transmission of S. aureus in non-human primates and their living environment, we conducted this study on all the non-human primates in a zoo park located in eastern China. Fecal samples from non-human primates and samples from their living environments were selectively enriched and conducted for S. aureus isolation. The whole genome analysis showed that these two MRSA t034/CC398 isolates were found from a monkey fecal sample and its living environment. The virulence factor analysis showed that these two MRSA isolates carried virulence factors relating to the cell adherence, biofilm formation, toxins, and human-associated immune evasion cluster. The further phylogenetic analysis revealed that these two MRSA isolates have a close genomic relationship with the human-associated clinical isolates from China, which indicates a potential risk of bidirectional transfer of MRSA between monkeys and humans. ABSTRACT: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the important antibiotic resistant pathogens causing infections in humans and animals. The increasing observation of MRSA in wildlife species has raised the concern of its impact on animal health and the potential of zoonotic transmission. This study investigated the prevalence of S. aureus in fecal samples from non-human primates in a zoo located in Jiangsu, China, in which 6 out of 31 (19.4%) fecal samples, and 2 out of 14 (14.3%) indoor room floor swab samples were S. aureus-positive. The antibiotic susceptibility tests of the eight isolates showed that the two isolates were resistant to both penicillin and cefoxitin, the three isolates were resistant only to penicillin, while three isolates were susceptible to all detected antibiotics. The two isolates resistant to cefoxitin were further identified as MRSA by the presence of mecA. Five different spa types were identified including t034 of two MRSA isolates from Trachypithecus francoisi, t189 of two methicillin-susceptible S. aureus (MSSA) isolates from Rhinopithecus roxellana, t377 of two MSSA isolates from Colobus guereza, and two novel spa types t19488 and t19499 from Papio anubis. Whole genome sequencing analysis showed that MRSA t034 isolates belonged to ST398 clustered in clonal complex 398 (CC398) and carried the type B ΦSa3 prophage. The phylogenetic analysis showed that the two MRSA t034/ST398 isolates were closely related to the human-associated MSSA in China. Moreover, two MRSA isolates contained the virulence genes relating to the cell adherence, biofilm formation, toxins, and the human-associated immune evasion cluster, which indicated the potential of bidirectional transfer of MRSA between monkeys and humans. This study is the first to report MRSA CC398 from monkey feces in China, indicating that MRSA CC398 could colonize in monkey and have the risk of transmission between humans and monkeys.