AppendixA All the supplements can be accessed via: S1. Experimental details and bioinformation analysis for 16S rRNA amplicon sequencing and metagenomic sequencing; Figure S2. Differences in α diversity on four indices between the salt beach and nonsalt beach (Fig. 1 A – Shannon ndex; B – Simpson index; C – Ace index; D – Chao index). Figure S3. A. Distance box plot on the genus level for SAB and NSB groups by ANOSIM analysis. B. Distance box plot on OTU level for SAB and NSB groups by ANOSIM analysis Figure S4. A – Source tracking of faecal contamination of two marine beaches; B – Unique common genera between beaches and warm-blooded hosts. Figure S5. A – Community heat map analysis for beaches and hosts at the species level. B – Community heatmap analysis for beaches and hosts at the genus level. Supplementary Materials. S1 – experiment methods and bioinformation analysis for 16S rRNA amplicon sequencing and metagenomic sequencing. S2 – differences in α diversity on four indices between the salt beach and non-salt beach (Fig. 1 A: Shannon index; B: Simpson index; C: Ace index; D: Chao index). S3 – Figure A, B, C, and D. ANOSIM analysis of the distance box plot on the genus and OTU levels between SAB and NSB, as well as within the SAB group. S4 – Figure A. Source tracking of faecal contamination of two marine beaches; B. Unique common genera between beaches and warm-blooded hosts. S5 – Figure A. Community heat map analysis for beaches and hosts at species level. B – community heat map analysis for beaches and hosts at genus level.
The aim of the study was to explore the correlation between characteristics of microbial community, pathogenic bacteria and high-risk antibiotic-resistant genes, between coastal beaches and a multi-warm-blooded host, as well as to determine potential species biomarkers for faecal source contamination on tropical coastal beaches in China.

Material and methods:
The ‘One-Health’ approach was used in a microbiological study of beaches and warm-blooded hosts. The was analyzed using 16S rRNA gene amplicons and shotgun metagenomics on Illumina NovaSeq.

The Chao, Simpson, Shannon, and ACE indices of non-salt beach were greater than those of salt beaches at the genus and OTU levels (P < 0.001). Bacteroidota, Halanaerobiaeota, Cyanobacteria, and Firmicutes were abundant on salt beaches (P<0.01). Human-sourced microorganisms were more abundant on salt beaches, which accounted for 0.57%. Faecalibacterium prausnitzii and Eubacterium hallii were considered as reliable indicators for the contamination of human faeces. High-risk carbapenem-resistant Klebsiella pneumoniae and the genotypes KPC-14 and KPC-24 were observed on salt beaches. Tet(X3)/tet(X4) genes and four types of MCR genes co-occurred on beaches and humans; MCR9.1 accounted for the majority. Tet(X4) found among Cyanobacteria. Although rarely reported at Chinese beaches, pathogens, such as Vibrio vulnificus, Legionella pneumophila, and Helicobacter pylori, were observed.

The low microbial community diversity, however, did not indicate a reduced risk. The transfer of high-risk ARGs to extreme coastal environments should be given sufficient attention.

The research was funded by the Science and Technology Department of Hainan Province (Grant No. ZDYF2020181), and the National Natural Science Foundation of China (Grant No. 81460487). The authors particularly express their thanks to the participants in Danzhou County for their invaluable collaboration and support.
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