AUTHOR=Limayem Alya , Wasson Sarah , Mehta Mausam , Pokhrel Anaya Raj , Patil Shrushti , Nguyen Minh , Chen Jing , Nayak Bina 

TITLE=High-Throughput Detection of Bacterial Community and Its Drug-Resistance Profiling From Local Reclaimed Wastewater Plants

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=9

YEAR=2019

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2019.00303

DOI=10.3389/fcimb.2019.00303

ISSN=2235-2988

ABSTRACT=<p>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 (<italic>Enterococcus faecium</italic> and <italic>Streptococcus surrogates, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa</italic>, and <italic>Enterobacter</italic> species along with the emerging nosocomial <italic>Escherichia</italic> 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, <italic>Pseudomonas</italic> was the most predominant genus related to ESKAPE in influent, with sequence reads corresponding to 21.356%, followed by <italic>Streptococcus</italic> (6.445%), <italic>Acinetobacter</italic> (0.968%), <italic>Enterococcus</italic> (0.063%), <italic>Klebsiella</italic> (0.038%), <italic>Escherichia</italic> (0.028%) and <italic>Staphylococcus</italic> (0.004%). Despite the different treatment methods used, the effluent still revealed the presence of some <italic>Pseudomonas</italic> strains (0.066%), and a wide range of gram-positive cocci, including <italic>Staphylococcus</italic> (0.194%), <italic>Streptococcus</italic> (0.63%) and <italic>Enterococcus</italic> (0.037%), in addition to gram-negative <italic>Acinetobacter</italic> (0.736%), <italic>Klebsiella</italic> (0.1%), and <italic>Escherichia</italic> sub-species (0.811%). The AST results indicated that the strains <italic>Escherichia</italic> along with <italic>Klebsiella</italic> and <italic>Acinetobacter</italic>, isolated from the effluent, displayed resistance to 11 antibiotics, while <italic>Pseudomonas</italic> was resistant to 7 antibiotics, and <italic>Streptococcus</italic> along with <italic>Staphylococcus</italic> 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.</p>