Draft Genome Sequences of Three Multidrug-Resistant <named-content content-type='genus-species'>Staphylococcus</named-content> spp. Isolated from Hospital Wastewater in Malaysia

Staphylococcus spp. are Gram-positive bacteria that reside within the normal microbiota of humans and animals but pose a health threat as reservoirs of antimicrobial resistance genes. Here, we present the draft genome sequences of three Staphylococcus sp. strains isolated from hospital wastewater in Malaysia that demonstrated resistance to multiple antibiotics. S taphylococcus spp. have emerged as a major health threat in nosocomial and community settings due to their active participation in the transmission of antibiotic resistance (1–3). Water contaminated with antibiotics from human waste, livestock farms, health care settings, and pharmaceutical residues may serve as a risk factor that places Southeast Asia at the highest risk for the emergence and spread of antibiotic resistance in humans (4–6). Here, we isolated and sequenced three Staphylococcus sp. strains from hospital wastewater to discover the mechanisms and mobile genetic elements that are responsible for the transmission of resistance genes in the tropics. Wastewater samples were collected from a river near the sewage treatment plant of the University of Malaya Medical Centre (UMMC) (3°07901.50N, 101°39951.40E) in Kuala Lumpur, Malaysia. The samples were filtered and grown on mannitol salt agar (MSA) at 37°C for 24 h, followed by Gram staining and catalase and coagulase tests (7). The genus and species were confirmed after 16S rRNA gene sequencing using the 27F and 1492R primers, as well as the nuc primer (which encodes a thermonuclease enzyme in certain Staphylococcus aureus isolates). Isolates were then screened for multidrug-resistant (MDR) strains using the disk diffusion assay (8, 9). Staphylococcus sp. strains S36, S59, and S75 showed resistance to multiple classes of antibiotics and were selected for whole-genome sequencing. Genomic DNA of the strains was extracted from pure cultures grown overnight in nutrient broth using the HiYield genomic DNA minikit (RBC BioScience). A 400-bp library was constructed using the Ion Xpress Plus fragment library kit (Thermo Fisher Scientific) according to the manufacturer’s protocol and quantified using an Agilent 2100 bioanalyzer. The library was diluted prior to template preparation using the Ion Chef system (Thermo Fisher Scientific), followed by sequencing using the Ion S5XL system (Thermo Fisher Scientific). Torrent Suite software (Thermo Fisher Scientific) was used for raw data analysis, alignment, and variant calling. Short reads from the Ion S5XL system were quality trimmed and assembled using CLC Genomics Workbench v11.0. 1. The reads were trimmed with the following parameters: quality score limit, 0.05; discarded reads, ,400 nucleotides; and maximum number of ambiguous nucleotides, 2. Default parameters were used for the assembly. Citation Ahmad Sabri NS, Mohd Zulkeflle SN, Yusof N, Md Akhir FN, Othman N, Zakaria Z, Hara H. 2021. Draft genome sequences of three multidrug-resistant Staphylococcus spp. isolated from hospital wastewater in Malaysia. Microbiol Resour Announc 10:e00332-21. https://doi.org/10.1128/MRA.00332-21. Editor Steven R. Gill, University of Rochester School of Medicine and Dentistry Copyright © 2021 Ahmad Sabri et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Address correspondence to Hirofumi Hara, hhara@mjiit.jp. Received 2 April 2021 Accepted 11 April 2021 Published 6 May 2021 Volume 10 Issue 18 e00332-21 mra.asm. org 1 GENOME SEQUENCES