Data_Sheet_1_High-Throughput Detection of Bacterial Community and Its Drug-Resistance Profiling From Local Reclaimed Wastewater Plants.docx

Treated wastewater from reclaimed facilities (WWTP) has become a reusable source for a variety of applications, such as agricultural irrigation. However, it is also a potential reservoir of clinically-relevant multidrug resistant (MDR) pathogens, including ESKAPE (Enterococcus faecium and Streptococcus surrogates, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species along with the emerging nosocomial Escherichia strains). This study was performed to decipher the bacterial community structure through Illumina high throughput 16S rRNA gene sequencing, and to determine the resistance profile using the Sensititre antimicrobial susceptibility test (AST) conforming to clinical lab standards (NCCLS). Out of 1747 bacterial strains detected from wastewater influent and effluent, Pseudomonas was the most predominant genus related to ESKAPE in influent, with sequence reads corresponding to 21.356%, followed by Streptococcus (6.445%), Acinetobacter (0.968%), Enterococcus (0.063%), Klebsiella (0.038%), Escherichia (0.028%) and Staphylococcus (0.004%). Despite the different treatment methods used, the effluent still revealed the presence of some Pseudomonas strains (0.066%), and a wide range of gram-positive cocci, including Staphylococcus (0.194%), Streptococcus (0.63%) and Enterococcus (0.037%), in addition to gram-negative Acinetobacter (0.736%), Klebsiella (0.1%), and Escherichia sub-species (0.811%). The AST results indicated that the strains Escherichia along with Klebsiella and Acinetobacter, isolated from the effluent, displayed resistance to 11 antibiotics, while Pseudomonas was resistant to 7 antibiotics, and Streptococcus along with Staphylococcus were resistant to 9 antibiotics. Results herein, proved the existence of some nosocomial MDR pathogens, known for ESKAPE, with potential drug resistance transfer to the non-pathogen microbes, requiring targeted remediation.

经过处理的重水厂（WWTP）废水已成为多种应用的可再生资源，如农业灌溉。然而，它也是临床相关多药耐药（MDR）病原体的潜在储存库，包括ESKAPE（肠球菌、替代链球菌、金黄色葡萄球菌、肺炎克雷伯菌、鲍曼不动杆菌、铜绿假单胞菌和肠杆菌属，以及新兴的医院感染大肠杆菌菌株）。本研究旨在通过Illumina高通量16S rRNA基因测序解析细菌群落结构，并使用符合临床实验室标准（NCCLS）的Sensititre抗菌药物敏感性测试（AST）确定耐药谱。从废水和出水口检测到的1747个细菌菌株中，铜绿假单胞菌是出水口中最主要的与ESKAPE相关的属，其序列读数占21.356%，其次是链球菌（6.445%）、不动杆菌（0.968%）、肠球菌（0.063%）、克雷伯菌（0.038%）、大肠杆菌（0.028%）和葡萄球菌（0.004%）。尽管使用了不同的处理方法，但出水仍然检测到一些铜绿假单胞菌菌株（0.066%），以及包括葡萄球菌（0.194%）、链球菌（0.63%）和肠球菌（0.037%）在内的广泛革兰氏阳性球菌，以及革兰氏阴性不动杆菌（0.736%）、克雷伯菌（0.1%）和大肠杆菌亚种（0.811%）。AST结果显示，从出水口分离的大肠杆菌、克雷伯菌和不动杆菌菌株对11种抗生素具有耐药性，铜绿假单胞菌对7种抗生素具有耐药性，而链球菌和葡萄球菌对9种抗生素具有耐药性。本研究结果证实了一些医院感染的多药耐药病原体（ESKAPE）的存在，这些病原体可能将药物耐药性转移到非病原微生物，需要针对性的修复措施。