Molecular Characterization of Staphylococcus aureus Complex Isolated from Free-Ranging Long-Tailed Macaques at Kosumpee Forest Park, Maha Sarakham, Thailand.

The Staphylococcus (S.) aureus complex, including methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA), and S. argenteus are bacterial pathogens that are responsible for both human and animal infection. However, insights into the molecular characteristics of MRSA, MSSA, and S. argenteus carriages in wildlife, especially in long-tailed macaques, rarely have been reported in Thailand. The objective of this study was to assess molecular characterization of MRSA, MSSA, and S. argenteus strains isolated from free-ranging long-tailed macaques (Macaca fascicularis) at Kosumpee Forest Park, Maha Sarakham, Thailand. A total of 21 secondary bacterial isolates (including 14 MRSA, 5 MSSA, and 2 S. argenteus) obtained from the buccal mucosa of 17 macaques were analysed by a Polymerase chain reaction (PCR) to identify several virulence genes, including pvl, tst, hla, hlb clfA, spa (x-region), spa (IgG biding region), and coa. The most prevalent virulence genes were clfA, coa, and the spa IgG biding region which presented in all isolates. These data indicated that MRSA, MSSA, and S. argenteus isolates from the wild macaques at Kosumpee Forest Park possess a unique molecular profile, harbouring high numbers of virulence genes. These findings suggest that wild macaques may potentially serve as carriers for distribution of virulent staphylococcal bacteria in the study area.

Investigation of methicillin-resistant Staphylococcus aureus, methicillin-susceptible Staphylococcus aureus, and Staphylococcus argenteus from wild long-tailed macaques (Macaca fascicularis) at Kosumpee Forest Park, Maha Sarakham, Thailand.

Background and Aim: In the past, the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) infections in both humans and animals has increased across Thailand. Staphylococcus argenteus has been associated with infections among humans, exotic pets, and livestock. Both species have been identified in non-human primate species from geographically diverse locations but not from non-human primates in Thailand. This study aimed to determine the presence of MRSA/methicillin-susceptible S. aureus (MSSA) and S. argenteus isolates collected from buccal swab samples in Macaca fascicularis at Kosumpee Forest Park (KFP), Maha Sarakham, Northeast Thailand. Materials and Methods: Aseptic buccal swab samples were collected from 30 free-ranging macaques in November 2018. All isolates were tested using multiple biochemical tests and S. aureus latex slide agglutination test. Presumptive S. aureus isolates were tested for the presence of the mecA gene using polymerase chain reaction (PCR) assays. The isolates were phenotypically determined to be resistant to a ß-lactam antibiotic using the disk diffusion method with a 30 mg cefoxitin disk. The isolates were analyzed by PCR for the non-ribosomal peptide synthetase (NRPS) gene to distinguish S. argenteus from S. aureus. Results: Fifteen macaques (50%) were colonized with S. aureus and 21 isolates were characterized. Three of the macaques carried both the MRSA and MSSA isolate. One animal carried both MRSA and S. argenteus isolate, and one animal carried only S. argenteus. The NRPS gene analysis confirmed that 2 isolates (9.52%) were S. argenteus and 19 isolates (90.48%) were S. aureus [five MSSA and 14 MRSA]. Conclusion: This study is the first to identify MRSA/MSSA and S. argenteus in wild free-ranging M. fascicularis from Thailand at the KFP in Maha Sarakham. This study is also the first report on the occurrence of S. argenteus carriage in M. fascicularis from Thailand.

Leptospira Seroprevalence in Free-Ranging Long-Tailed Macaques (Macaca fascicularis) at Kosumpee Forest Park, Maha Sarakham, Thailand.

BACKGROUND: Leptospirosis is a zoonotic disease that is ubiquitously distributed and is classified as a re-emerging infectious disease in humans and animals. Many serovars are carried by wildlife; all of them are capable of causing illness in humans. The purpose of this study was to investigate the prevalence of Leptospirosis in wild long-tailed macaques (Macaca fascicularis) at Kosumpee Forest Park, Mahasarakham, Thailand. METHODS: A cross-sectional study was conducted at the park. Blood samples were collected via saphenous vein from 30 free-ranging long-tailed macaques. All samples were tested by the microscopic agglutination test. The LipL32 gene was used to detect pathogenic Leptospira in blood samples by conventional polymerase chain reaction. RESULTS: Screening of the 30 wild macaques showed an overall Leptospira seroreactivity of 13.33%. Three of 30 macaques reacted against Leptospira serovar Shermani and one macaque was infected with Leptospira serovar Sejroe. None of the macaques presented clinical signs of leptospirosis. None of the blood samples showed the detection of the LipL32 gene. CONCLUSIONS: The results indicate that the long-tailed macaques at Kosumpee Forest Park may act as natural reservoirs for Leptospirosis. Further, the results provide evidence-based information indicating that several pathogenic Leptospira serovars are circulating in the wild macaques in the study area.

Correction: Fecal microbiota dysbiosis in macaques and humans within a shared environment.

[This corrects the article DOI: 10.1371/journal.pone.0210679.].

Fecal microbiota dysbiosis in macaques and humans within a shared environment.

Traditional zoonotic disease research focuses on detection of recognized pathogens and may miss opportunities to understand broader microbial transmission dynamics between humans, animals, and the environment. We studied human-macaque microbiome overlap in Kosum Phisai District, Maha Sarakham Province, Thailand, where a growing population of long-tailed macaques (Macaca fascicularis) in Kosumpee Forest Park interact with humans from an adjacent village. We surveyed workers in or near the park with elevated exposure to macaques to characterize tasks resulting in exposure to macaque feces in addition to dietary and lifestyle factors that influence gut microbiome composition. Fecal samples were collected from 12 exposed workers and 6 controls without macaque exposure, as well as 8 macaques from Kosumpee Forest Park and 4 from an isolated forest patch with minimal human contact. The V4 region of the 16S rRNA gene from fecal sample extracted DNA was amplified and sequenced using Illumina MiSeq to characterize the microbial community. A permuted betadisper test on the weighted UniFrac distances revealed significant differences in the dispersion patterns of gut microbiota from exposed and control macaques (p = 0.03). The high variance in gut microbiota composition of macaques in contact with humans has potential implications for gut microbiome stability and susceptibility to disease, described by the Anna Karenina principle (AKP). Human samples had homogenous variance in beta diversity but different spatial medians between groups (p = 0.02), indicating a shift in microbial composition that may be explained by fundamental lifestyle differences between the groups unrelated to exposure status. SourceTracker was used to estimate the percent of gut taxa in exposed humans that was contributed by macaques. While one worker showed evidence of elevated contribution, the overall trend was not significant. Task observations among workers revealed opportunities to employ protective measures or training to reduce exposure to occupational hazards. These results suggest the potential for hygiene measures to mitigate negative aspects of contact between humans and macaques in order to optimize the health of both populations.

Long-Tailed Macaques (Macaca fascicularis) in Urban Landscapes: Gastrointestinal Parasitism and Barriers for Healthy Coexistence in Northeast Thailand.

Gastrointestinal parasites have diverse life cycles that can involve people, animals, and the environment (e.g., water and soil), demonstrating the utility of One Health frameworks in characterizing infection risk. Kosumpee Forest Park (Thailand) is home to a dense population of long-tailed macaques (Macaca fascicularis) that frequently interact with tourists and local residents. Our study investigated the presence of zoonotic parasites, and barriers to healthy coexistence by conducting stool analysis on macaques (N = 102) and people (N = 115), and by examining risk factors for infection with a household questionnaire (N = 95). Overall, 44% of macaques and 12% of people were infected with one or more gastrointestinal helminths, including Strongyloides spp., Ascaris spp., and Trichuris sp. An adults-only generalized linear mixed model identified three factors significantly associated with human infection: household size, occupational exposure, and contact with macaque feces at home. Participants identified both advantages and disadvantages to living in close contact with macaques, suggesting that interventions to improve human and animal health in Kosumpee Forest Park would be welcome.

PREVIOUSLY UNREPORTED POPULATION OF RHESUS MACAQUES Macaca mulatta IN CHIANG RAI PROVINCE, THAILAND: PRELIMINARY OBSERVATIONS.

Efforts to document the distribution of macaques in Thailand have been gradually improving over the past several years. Here we add to the growing database with a summary of a previously unreported population of Rhesus Macaque Macaca mulatta located at Wat Phrabuddhabat Pa Reau in Chiang Rai Province. This work is part of an on-going study addressing human-primate conflict and coexistence in Thailand. The Wat covers an area of about 53 ha consisting of forest and ponds surrounded by an agricultural/rural residential area. The macaques receive some provisioning by the monks, nuns, local residents and occasional tourists. We conducted observations of the macaques and queried the monks, nuns and local residents during 20-21 November 2015 and 22-23 July 2016. Those queried reported population sizes ranging between "100" and "2000" monkeys and 1-4 groups. Based on our observations, we identified at least two groups of approximately 55 and 44 monkeys. All individuals queried reported crop raiding and expressed concern over an "increasing" monkey population and need for effective population management. We plan to follow up with a more intensive survey of this population to better assess conservation concerns, human-primate interaction, and options for healthy coexistence.