A New Source and Large Quantity of Resveratrol in Cratoxylum Species and Their Activities on Normal Human and Cancer Cells.

Cratoxylum formosum ssp. formosum (Cff), C. formosum ssp. pruniflorum (Cfp), and C. sumatranum (Cs) were investigated for phytochemical analysis. Toxicity testing, programmed cell death, and cell cycle arrest were tested on CHL-1, HCT-116, and HepG2 cancer cell lines, and human normal PBMCs. The results are revealed in the following order. The phytochemical percentages varied in each species, the quantity and concentration of α-amyrin and resveratrol were 0.038 mg/g and 0.955 mg/mL, and 0.064 mg/g and 0.640 mg/mL. The most studied Cratoxylum extracts showed IC50 values in PBMCs and cancer cell lines except for the hexane Cff and ethanol Cfp extracts. All studied extracts did not induce DNA breaks in PBMCs but caused significant DNA breaks in the cancer cell lines. All studied extracts induced both apoptosis and necrosis in cancer cell lines, and the DNA quantity in the S and G2-M phases decreased significantly but did not induce apoptosis and necrosis in PBMCs. Except for the ethanolic extracts of Cff and Cfp that induced PBMCs apoptosis and necrosis, these data confirmed that the three studied Cratoxylum samples have inhibiting properties for the growth of cancer cells and low toxicity to PBMCs. Cs showed more toxicity to cancer cell lines than Cf and cisplatin.

The chloroplast genome sequences of Ipomoea alba and I. obscura (Convolvulaceae): genome comparison and phylogenetic analysis.

Ipomoea species have diverse uses as ornamentals, food, and medicine. However, their genomic information is limited; I. alba and I. obscura were sequenced and assembled. Their chloroplast genomes were 161,353 bp and 159,691 bp, respectively. Both genomes exhibited a quadripartite structure, consisting of a pair of inverted repeat (IR) regions, which are separated by the large single-copy (LSC) and small single-copy (SSC) regions. The overall GC content was 37.5% for both genomes. A total of 104 and 93 simple sequence repeats, 50 large repeats, and 30 and 22 short tandem repeats were identified in the two chloroplast genomes, respectively. G and T were more preferred than C and A at the third base position based on the Parity Rule 2 plot analysis, and the neutrality plot revealed correlation coefficients of 0.126 and 0.105, indicating the influence of natural selection in shaping the codon usage bias in most protein-coding genes (CDS). Genome comparative analyses using 31 selected Ipomoea taxa from Thailand showed that their chloroplast genomes are rather conserved, but the presence of expansion or contraction of the IR region was identified in some of these Ipomoea taxa. A total of five highly divergent regions were identified, including the CDS genes accD, ndhA, and ndhF, as well as the intergenic spacer regions psbI-atpA and rpl32-ccsA. Phylogenetic analysis based on both the complete chloroplast genome sequence and CDS datasets of 31 Ipomoea taxa showed that I. alba is resolved as a group member for series (ser.) Quamoclit, which contains seven other taxa, including I. hederacea, I. imperati, I. indica, I. nil, I. purpurea, I. quamoclit, and I. × sloteri, while I. obscura is grouped with I. tiliifolia, both of which are under ser. Obscura, and is closely related to I. biflora of ser. Pes-tigridis. Divergence time estimation using the complete chloroplast genome sequence dataset indicated that the mean age of the divergence for Ipomoeeae, Argyreiinae, and Astripomoeinae, was approximately 29.99 Mya, 19.81 Mya, and 13.40 Mya, respectively. The node indicating the divergence of I. alba from the other members of Ipomoea was around 10.06 Mya, and the split between I. obscura and I. tiliifolia is thought to have happened around 17.13 Mya. The split between the I. obscura accessions from Thailand and Taiwan is thought to have taken place around 0.86 Mya.

The complete chloroplast genome of Pandanus amaryllifolius Roxb. ex Lindl. (Pandanaceae) and its phylogenetic relationship.

Pandanus amaryllifolius of Pandanaceae, a plant native to Southeast Asia, has been domesticated for its health benefits and aromatic leaves. It is also used for phytoremediation and soil rehabilitation. However, genetic studies of this species are limited. This study aims to expand its genomic information by assembling and characterizing the complete chloroplast genome of P. amaryllifolius. The chloroplast genome, which was 157,839 bp long, contains a total of 133 genes, including 87 protein-coding (CDS), 38 tRNA, and eight rRNA genes. The overall G/C content was 37.7%. A phylogenetic analysis using 79 shared unique CDS revealed a monophyletic relationship in Pandanales. Based on the limited sampling size, Pandanus amaryllifolius was the first to diverge in Pandanaceae. The genomic data will be useful for future phylogenetic and evolutionary studies of Pandanaceae.

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.

Complete Chloroplast Genome of Four Thai Native Dioscorea Species: Structural, Comparative and Phylogenetic Analyses.

The chloroplast genomes of Dioscorea brevipetiolata, D. depauperata, D. glabra, and D. pyrifolia are 153,370-153,503 bp in size. A total of 113 genes were predicted, including 79 protein-coding sequences (CDS), 30 tRNA, and four rRNA genes. The overall GC content for all four species was 37%. Only mono-, di-, and trinucleotides were present in the genome. Genes adjacent to the junction borders were similar in all species analyzed. Eight distinct indel variations were detected in the chloroplast genome alignment of 24 Dioscorea species. At a cut-off point of Pi = 0.03, a sliding window analysis based on 25 chloroplast genome sequences of Dioscorea species revealed three highly variable regions, which included three CDS (trnC, ycf1, and rpl32), as well as an intergenic spacer region, ndhF-rpl32. A phylogenetic tree based on the complete chloroplast genome sequence displayed an almost fully resolved relationship in Dioscorea. However, D. brevipetiolata, D. depauperata, and D. glabra were clustered together with D. alata, while D. pyrifolia was closely related to D. aspersa. As Dioscorea is a diverse genus, genome data generated in this study may contribute to a better understanding of the genetic identity of these species, which would be useful for future taxonomic work of Dioscorea.

Characterization of the plastid genome of Cratoxylum species (Hypericaceae) and new insights into phylogenetic relationships.

To expand the genomic information of Hypericaceae, particularly on Cratoxylum, we characterized seven novel complete plastid genomes (plastomes) of five Cratoxylum and two of its allied taxa, including C. arborescens, C. formosum subsp. formosum, C. formosum subsp. pruniflorum, C. maingayi, C. sumatranum, Hypericum hookerianum, and Triadenum breviflorum. For Cratoxylum, the plastomes ranged from 156,962 to 157,792 bp in length. Genomic structure and gene contents were observed in the five plastomes, and were comprised of 128-129 genes, which includes 83-84 protein-coding (CDS), 37 tRNA, and eight rRNA genes. The plastomes of H. hookerianum and T. breviflorum were 138,260 bp and 167,693 bp, respectively. A total of 110 and 127 genes included 72 and 82 CDS, 34 and 37 tRNA, as well as four and eight rRNA genes. The reconstruction of the phylogenetic trees using maximum likelihood (ML) and Bayesian inference (BI) trees based on the concatenated CDS and internal transcribed spacer (ITS) sequences that were analyzed separately have revealed the same topology structure at genus level; Cratoxylum is monophyletic. However, C. formosum subsp. pruniflorum was not clustered together with its origin, raising doubt that it should be treated as a distinct species, C. pruniflorum based on molecular evidence that was supported by morphological descriptions.

Chloroplast Genome Analysis for Genetic Information and Authentication in Five Barleria Species.

In order to authenticate the genomic information of Barleriacristata L., B. lupulina Lindl., B. repens Nees, B. siamensis Craib, and B. strigosa Willd, cp genomes were investigated. They revealed a general structure with a total size of 151,997-152,324 bp. The genomes encoded a total of 131 genes, including 86 CDS, 37 tRNA, and 8 rRNA genes. Other details found were as follows: different numbers and types of SSRs; identical gene content, which is adjacent to the border regions, except for B. strigosa, that revealed a shorter ndhF gene sequence and lacked the ycf1 gene; slightly different genetic distance values, which can be used for species identification; three distinct gaps of nucleotide variations between the species located at the intergenic spacer regions of the LSC and CDS of the SSC; three effective molecular markers derived from divergent hotspot regions, including the ccsA-ndhD, ndhA-ndhH-rps15, and ycf1. The genetic relationships derived from the cp genome and the CDS phylogenetic trees of Barleria and the 13 genera in Acanthaceae and different families, Scrophulariaceae and Phrymaceae, showed similar results. The six Barleria species as monophyletic groups with inner and outer outgroups were found to have perfect discrimination. These results have helped to authenticate the five Barleria species and the six genera in Acanthaceae.

Investigation of Morinda citrifolia Activities through Pinoresinol and α-EG Related Gene Expression.

α-EG is a unique substance that was first found in the leaves and fruits of Morinda citrifolia (Mc) growing in Thailand using GC-MS at 52.33% and 54.12%. It was then concentrated and its abundance quantified, along with that of pinoresinol, via GC, compared to the standards in leaves, ufp, rfp, rawfs, and seeds. α-EG and pinoresinol, which have collagen stimulating, skin whitening, and an inhibitory effect on wrinkle formation, were found in different concentrations and amounts. Three different concentrations of the five Mc part extracts were tested on NHDF for gene expression related to the aforementioned activities, COL1A1, COL1A2, and COL3A1, FGF1 and FGF7 by qRT-PCR. The results showed various expression levels, both stimulatory and inhibitory, with different concentrations of plant parts and genes. Similar results were revealed when the experiments were performed with Morus alba (Ma), which was found to contain 20.48 g protein p/100 g leaves at concentrations of 3.11 mg/mL. The studied Mc parts seem to have advantages based on the stated objectives, gene type and level of activity of each plant part. Rawfs and leaves supplemented with Ma samples were selected for toxicity tests with PBMCs. The lack of both cell and DNA toxicity from the rawfs indicated that they can be used safely.

Formulations for Effective Detoxification Derived from Three Medicinal Plants: Thunbergia laurifolia, Clerodendrum disparifolium and Rotheca serrata.

BACKGROUND: Based on the long history of the medicinal use of Thunbergia laurifolia, Clerodendrum disparifolium and Rotheca serrata, the extract formulations of these species: T. laurifolia and C. disparifolium; T. laurifolia and R. serrata; and T. laurifolia, C. disparifolium and R. serrata, called formulas 1, 2 and 3, were created for detoxification testing to take more advantage of each species. OBJECTIVE: The objective of this study is to estimate the detoxifying effects of studied extract formulations on human cell and tissue culture as a preclinical trial. METHODS: The major phytochemicals were derived by GC-MS. The detoxification efficacy of these formulations in cells and DNA levels were derived by MTT and comet assays in toxic PBMCs (incubated with rice whisky or bathroom cleaner). RESULTS: The phytochemical constituents were detected at 23.48% phytol and 43.03% oleamide in T. laurifolia; 12.88% oleamide, 20.93% 9,12,15-octadecatrien, 25.52% squalene, 22.19% butylated hydroxy toluene and 15.36% vitamin E in C. disparifolium; and 30.41% phytol, 32.78% oleamide, and 12.20%, 9,12,15-octadecatrien-1-ol in R. serrata. The toxic cells treated with the plant formulas 1, 2 and 3 showed no IC50 values, but formulas 1 and 2 displayed higher efficacies than formula 3 did. The comet assay indicated that the experiments (the treatment on toxic cells with the plant formulas) induced significant (p < 0.05) DNA damage compared to the negative control due to poisoning occurring before administration of the plant formulas. The OTM of the controls was significantly (p < 0.05) longer than the experimental samples showing significantly reduce the toxicity of the created formulations. CONCLUSION: The formulas showed high detoxification efficacies and formulations 1 and 2 resulted in higher levels of detoxification than formulation 3, especially in immediate treatment after receiving toxic substances.

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.

The Unique Substance, Lidocaine and Biological Activity of the Dioscorea Species for Potential Application as a Cancer Treatment, Natural Pesticide and Product.

The six Dioscorea species, D. brevipetiolata, D. bulbifera, D. depauperata (Dd), D. glabra (Dg), D. pyrifolia and D. hamiltonii were analyzed for phytochemicals, toxicity in PBMCs, and biological activity in two cancer cell lines by MTT and comet assays, and pesticide efficiency. Via GC-MS, lidocaine was found to be the predominant compound in two of the studied species. To confirm the systematics, lidocaine was also found in lower amounts in 11 species. The MTT assay showed no toxicity in all six of the studied species. The comet assay showed the key result that the ethanol extracts of Dd and Dg violently broke DNA into pieces. Biological activity of these two species' extracts showed toxicity on HepG2 and no effects on HCT-116. The water extracts of Dd and Dg, applied to Brassica chinensis showed high efficiency as a bioprotectant. In summary, lidocaine seems to be the predominant identifying compound of the genus Dioscorea in Thailand, which is useful in systematics. At least the two species, Dd and Dg, may be used for human hepatocyte cancer treatment and as an alternative pesticide for economically important vegetables. Dioscorea species containing lidocaine or extracted lidocaine have promise for natural product creation.

One Health Approach of Melioidosis and Gastrointestinal Parasitic Infections from Macaca fascicularis to Human at Kosumpee Forest Park, Maha Sarakham, Thailand.

BACKGROUND AND OBJECTIVES: Gastrointestinal parasitic and melioidosis infections are major causes of morbidity and mortality from infectious disease in rural areas, especially in northeastern Thailand. Both diseases are zoonotic giving rise to health problems in both long-tailed macaques and in humans. In Thailand, macaques have adapted to live and share space with humans and can spread some zoonoses to humans. Therefore, this research aimed to measure the prevalence of gastrointestinal parasitic infections and melioidosis in long-tailed macaques at Kosumpee Forest Park and measure associated risk factors of their diseases among people in this area. METHODS: This study was conducted at Kosumpee Forest Park, Maha Sarakham, Thailand. Twenty-eight blood samples and 135 fecal samples were collected from free-ranging long-tailed macaques. Blood samples were tested by indirect hemagglutination test and fecal samples were analyzed by formalin-ethyl acetate concentration technique. A cross-sectional study was conducted among 350 respondents who were involved with the Forest Park using a multi-stage stratified random sampling method and performed to measure knowledge, attitude, and practice toward the zoonoses among the respondents. RESULTS: It was found that seroprevalence of melioidosis was 57.1% from macaque samples. The prevalence of gastrointestinal parasites infection was 35.11% from fecces samples, including Strongyloides spp. (15.27%), Trichuris spp. (22.9%), hookworm (4.58%) and Ascarid spp. (1.53%). KAP study indicated that the level of knowledge related to melioidosis and gastrointestinal parasites of people in the area was very low and moderate, respectively. The attitude of respondents who were aware of the diseases was at a moderate level for melioidosis and a high level for parasitic infection. CONCLUSION: The study therefore emphasizes the importance of one health approach for diagnosis, surveillance and management of zoonotic diseases to promote the development of hygiene measures and to educate people in the community around Kosumpee Forest Park.

Oleamide in Ipomoea and Dillenia Species and Inflammatory Activity Investigated through Ion Channel Inhibition.

BACKGROUND: Oleamide is an essential substance for human health. So, the plants with high oleamide content are great sources for health care products. OBJECTIVE: This study is conducted to investigate the quality of oleamide in plants and test the bioactivity in the selected two studied species. METHODS: The three Ipomoea and five Dillenia species including Ipomoea alba, Ipomoea aquatica and Ipomoea pes-caprae, and Dillenia indica, Dillenia obovata, Dillenia ovata, Dillenia parviflora and Dillenia pentagyna were investigated for the quantity of oleamide by high-performance liquid chromatography. The biological activity test was conducted on the powder formulation of the chosen plants, Dillenia ovata and Dillenia parviflora at a ratio of 30:70, for anti-inflammatory activity ex vivo on a panel of molecular targets through ion channel inhibition including voltage-gated sodium channel, voltage-gated potassium channel, and the cardiac ion as human ether-a-go-go related gene. RESULTS: The results showed that the leaf extracts of I. aquatica and D. ovata gave the highest and subsequent oleamide quantity i.e. 7.52 and 5.17 mg/g, respectively. Out of the Dillenia formulation which contained various compounds, oleamide showed the highest percentages of inhibition at 8.0-20.0%, and 6.2-14.2% in voltage-gated sodium channel, and voltage-gated potassium channel which had slightly lower values than the oleamide standard, and no effect as 0.0% value inhibition in the cardiac ion channel. CONCLUSION: The Dillenia formulation exhibits anti-inflammatory activity without affecting the heart. Accordingly, the three studied Ipomoea and three studied Dillenia species may be used for the same activity as a single component or formulation with effective solvent for disease treatments.

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.

PRE-CLINICAL EVALUATION OF EXTRACTS AND ESSENTIAL OILS FROM BETEL-LIKE SCENT PIPER SPECIES IDENTIFIED POTENTIAL CANCER TREATMENT.

BACKGROUND: Nine Piper species with betel-like scents are sources of industrial and medicinal aromatic chemicals, but there is lack of information on cytotoxicity and genotoxicity for human safety, including how these plants impact human cervical cancer cell line. METHODS: Plant leaves were extracted with hexane and hydro-distilled for essential oils. The extracts and oils were pre-clinically studied based on cyto - and genotoxicity using microculture tetrazolium (MTT) and comet assays. RESULTS: The crude extracts showed an IC50 in leukocytes and HeLa cells of 58.59-97.31 mg/ml and 34.91-101.79 mg/ml, the LD50 is higher than 5000 mg/kg. With lower values than the crude extracts, the essential oils showed an IC50 in leukocytes and HeLa cells of 0.023-0.059 µg/ml and 0.025-0.043 µg/ml the LD50 is less than 50 mg/kg. IC50 values showed that the essential oils were highly toxic than the crude extracts. At the level of human genetic materials, the crude extracts of two species, including P. betloides and P. crocatum, showed a significant toxicity (p < 0.05) in leukocytes. The other samples were non-toxic. The crude extracts of all samples showed significant genotoxicity in HeLa cells. The essential oils of all studied Piper species showed insignificant toxicity in leukocytes. For HeLa cells, the eight-studied species showed significant toxicity in HeLa cells, whereas only P. submultinerve showed insignificant toxicity. CONCLUSION: The crude extracts and essential oils should be tested as putative cervical cancer treatments due to less toxicity in human normal cells.

Major Phytochemical as γ-Sitosterol Disclosing and Toxicity Testing in Lagerstroemia Species.

Medicinal plants in genus Lagerstroemia were investigated for phytochemical contents by GC-MS and HPLC with ethanol and hexane extracts and their toxicity MTT and comet assay on human peripheral blood mononuclear cells (PBMCs). γ-Sitosterol is the major component found in all species at 14.70-34.44%. All of the extracts, except for L. speciosa ethanol extract, showed high percentages of cell viability. The IC50 value, 0.24 mg/mL, of ethanol L. speciosa extract predicted an LD50 of 811.78 mg/kg, which belongs to WHO Class III of toxic chemicals. However, in-depth toxicity evaluation by the comet assay showed that the four tested species induced significant (p < 0.05) DNA damage in PBMCs. γ-Sitosterol was previously reported to possess antihyperglycemic activity by increasing insulin secretion in response to glucose. Nonetheless, consumers should consider its toxicity, and the amount of consumption should be of concern.