Lab Matters Summer 2019 - Page 61

APHL 2019 POSTER ABSTRACTS New York City’s Drinking Water Microbiome — Microbial Diversity and Distribution C. Kretz 1 , J. Novak 2 , I. Rubinstein 2 , Y. Chekoff 2 , S. Silver 2 , T. Geiz 2 , S. Hughes 2 , A. Capetanakis 3 , S. Feud 3 , J. Rakeman 2 , E. Omoregie 2 ; 1 Centers for Disease Control and Prevention, 2 New York City Public Health Laboratory, 3 New York City Department of Environmental Protection Drinking water is one of our most valuable and closely monitored resources and its quality can have an immense influence on human health. Previous studies have reported that drinking water hosts diverse assemblages of microorganisms. This includes opportunistic pathogens that can host virulence and antimicrobial resistance genes, which are capable of many different types of environmental transformations and affect drinking water quality. New York City’s (NYC) water distribution system has a unique, comprehensive and efficient chemical and microbiological water quality monitoring program. This monitoring has shown the drinking water is high quality and free from enteric pathogens. However, relatively little is known about the microbial community composition in NYC’s drinking water. The project, which is ongoing, is to investigate the microbial community structure and metabolic functionality of NYC’s drinking water microbiome in relation to water source and treatment. This study coupled 16S rRNA gene sequencing, metagenomics sequence analysis, and physiological measurements. Samples were collected monthly over a 1-year period from 19 sites that included pretreated and posttreated waters. Preliminary data analysis of four months of 16S rRNA gene sequence revealed presence of more than 695 bacterial genera. Many of these genera are known for nutrient cycling in soil and aquatic systems, including Firmicutes, Azospira spp., Shewanella spp., and Methylobacter spp. Enteric pathogens, (e.g., E. coli or Salmonella) were not detected. However, genera that include opportunistic pathogens and are common in freshwater aquatic systems (e.g., Legionella spp. and Mycobacterium spp.) were detected. A comparison between microbial communities based on 16S rRNA sequences indicated that Actinobacteria sequences dominated the microbiome of pretreated waters by at 69% of the total number of sequences, while comprising only 5% from posttreated waters. Whole genome metagenomics sequencing and metabolic functionality is in the process of being assessed. Use of 16S rRNA gene sequencing indicates that NYC’s drinking water microbiome is diverse and replete with microorganisms common to other freshwater environments. This study provides a deeper understanding of the microbiome found in drinking water using metagenomic tools to investigate presence of potential pathogens, virulence factors, and antibiotic genes. It can provide new insights on potential biological processes associated with the microbiome and pathogen detection of drinking water microbial communities. The main goal is to ultimately protect public health by laying ground work for using new metagenomics tools for water surveillance. Presenter: Cecilia Kretz, Centers for Disease Control and Prevention, Atlanta, GA, Genetic Characterization of Human Adenoviruses Associated with Multiple Respiratory Outbreaks in the United States in 2018 X. Lu, S. Sakthivel, E. Schneider, H. Biggs, J. Watson, S. Gerber and S. Lindstrom, Centers for Disease Control and Prevention Background: Human adenoviruses (HAdVs) are medically important PublicHealthLabs @APHL pathogens responsible for substantial human disease burden. HAdV infections can cause a wide variety of clinical syndromes, including respiratory, ophthalmic, gastroenteric, neurologic and immuno- compromised host disease. HAdVs are classified into 7 species (A-G). Over 85 genotypes have been described by computational analysis of complete genomic sequences. Species B and E HAdVs, in particular types 3, 4, 7 and 14 are more commonly associated with outbreaks of severe acute respiratory infections. In 2018, CDC provided laboratory support to US public health laboratories to identify and characterize HAdVs associated with respiratory disease outbreaks and individual cases occurring in the US. Objective: To characterize HAdVs associated with human respiratory cases and outbreaks in multiple states in the US in 2018 using a comprehensive set of genetic analysis methods developed at CDC. Methods: Respiratory specimens collected from multiple HAdVs- associated respiratory disease cases in 2018 were sent to CDC for further characterization. Specimens were tested by a universal pan-HAdV real-time PCR (rPCR) assay to confirm HAdV detection and characterized by HAdV type-specific rPCR assays. Virus isolation and whole genome sequencing were performed to determine HAdV genotypes. DNA libraries were constructed using Nextera XT DNA Library Prep Kit and paired-end sequencing was performed on the Illumina MiSeq. De novo and reference-guided genome assemblies were achieved using CLC Genomics Workbench V11.0. Phylogenetic trees of the complete genome sequences were constructed by the neighbor-joining method implemented in MEGA V7. In silico restriction enzyme analysis was performed using Geneious 9.0.5. Results: HAdV-type specific rPCR identified four different HAdV types associated with respiratory disease clusters and cases: HAdV-3, HAdV-4, HAdV-7, HAdV-14, mixed HAdV-4 and HAdV-7. In silico restriction enzyme analyses identified HAdV-7 outbreak strains as genotype 7d that recently emerged in the US in 2013. HAdV-14 outbreak strains were identified as genotype 14p1. HAdV-4 strains were identified as 4a1 genotype. HAdV-3 strains were typed as genotype 3a2. Conclusions: In 2018, several US state public health laboratories reported cases and outbreaks of HAdV associated severe respiratory disease and CDC provided comprehensive genetic analysis that identified four different HAdVs types. Application of these genetic analysis methods permitted detailed characterization of the virus confirming its etiologic link with the cases and outbreaks. This demonstrates the value of rapid characterization of currently circulating HAdV types in supporting the public health response to control HAdV-associated outbreaks, and improving our understanding of the prevalence of respiratory illness associated with HAdVs. Presenter: Xiaoyan Lu, Centers for Disease Control and Prevention, Atlanta, GA,  Development of a Targeted Next-Generation Sequencing Method to Detect Drug Resistance in Mycobacterium tuberculosis E. Ransom, S. Burns, B. Metchock and J. Posey, Centers for Disease Control and Prevention Conventional drug susceptibility testing in Mycobacterium tuberculosis (Mtb) requires weeks-to-months to complete due to the slow growth rate of Mtb. Reducing the turnaround time is Summer 2019 LAB MATTERS 59