Lab Matters Summer 2018 - Page 66

APHL 2018 Annual Meeting Poster Abstracts of highly related isolates can help support the confirmation of specific sources of LD infection and inform the development of water management programs. We analyzed the genetic relatedness of environmental isolates recovered from 2 facilities (located in Georgia and Illinois) where clinical isolates from individuals with confirmed or suspected healthcare exposures were also available. In the Georgia facility, L. pneumophila Sequence Types (ST) 1 and 36 were recovered, however, available clinical isolates belonged to ST1. wgMLST analysis revealed that environmental ST1 isolates recovered from samples taken nearly 6 months apart and clinical isolates recovered in previous years shared >99% identical alleles. Similarly, ST1 and ST36 isolates were recovered from the Illinois facility, however, clinical isolates belonged to ST36. Genomic analysis of the clinical isolates and ST36 environmental isolates recovered from samples collected over 3 consecutive years revealed that the isolates shared >99% identical alleles. These data support the notion that some common L. pneumophila sequence types can persist in healthcare facilities and highlight the need for genome sequence analysis of isolates obtained from large, complex water systems in an effort to find ways to reduce the risk of Legionella growth and transmission. The findings and conclusions in this presentation are those of the authors and do not necessary represent the official position of the Centers for Disease Control and Prevention. cases in Ohio and Michigan. This novel clade belongs to sequence type 11/clonal complex 11, a hyperinvasive lineage that has caused meningococcal outbreaks. Further in-depth characterization found that the isolate possessed specific characteristics from urethritis- associated Nm genomes including multigene deletion at the capsule locus, causing its NG phenotype and the presence of the aniA gene, a gonococcal-acquired gene suspected to allow survival in anaerobic environments. Vaginal and nasopharyngeal swabs were collected from the mother to determine potential carriage and transmission, but no cultures were recovered. Our investigation demonstrates that an isolate belonging to the Nm urethritis clade has caused neonatal conjunctivitis. Modes of transmission are not fully understood for this clade and future investigations of Nm recovered from nonsterile sites (e.g., urogenital and oropharyngeal sites) for patients with non- invasive disease will be important to increase our understanding of non-invasive Nm. State and local health departments should consider investigating Nm conjunctivitis among newborns to more fully understand the clinical implications, modes of transmission and epidemiologic evolution of this emerging urethritis clade. Presenter: Brian Raphael, PhD, Centers for Disease Control and Prevention, Atlanta, GA, Phone: 404.639.4292, Email: braphael@cdc.gov Analysis of Microbial Diversity — A Novel Metagenomic Approach to Decipher Beach Microbiome Whole-Genome Sequencing Analysis of Neisseria meningitidis from the Urethritis Clade Causing Conjunctivitis in a Newborn Delivered by Cesarean — New York, August 2017 C. Kretz 1 , B. Genevieve 2 , M. Aldrich 3 , D. Bloch 2 , T. Halse 4 , Q. Liu 1 , E. Gonzalez 1 , E. Omoregie 1 , G. Zayas 2 , J. Wang 2 , M. Levi 3 , 5 , S. Hughes 1 , J. Rakeman 1 , D. Weiss 2 , K. Musser 4 ; 1 New York City Public Health Laboratory, New York, NY, 2 New York City Department of Health and Mental Hygiene, New York, NY, 3 Children’s Hospital at Montefiore, New York, NY, 4 Wadsworth Center, New York State Department of Health, Albany, NY, 5 Albert Einstein College of Medicine Neisseria meningitidis (Nm) is a leading cause of bacterial meningitis and is identified as a rare cause of newborn conjunctivitis linked to perinatal transmission from a genitally or nasopharyngeally colonized parent. Additionally, since 2015, it has been associated with outbreaks of urethritis. On August 31, 2017, Montefiore Medical Center isolated a nongroupable Nm (NmNG) from the eye of an infant aged 3 days with conjunctivitis. The infant was born to a healthy mother in a New York City (NYC) hospital, by cesarean section done due to prolonged rupture of membranes. For in-depth molecular characterization of the strain responsible for the disease and to determine the genetic similarity to circulating strains, we used whole-genome sequencing (WGS) and phylogenetic analysis. We used phylogenetic analyses to compare the sequences with publicly available Nm sequences and available NYC sequences collected for outbreaks and sporadic case investigations. Genomic analyses confirmed that the conjunctivitis isolate was phylogenetically part of the Nm urethritis clade with between 20-50 single nucleotide polymorphisms (SNP) differences. Moreover, the isolate possessed specific molecular characteristics found in the emerging clade of NmNG associated with an increase of urethritis 64 LAB MATTERS Summer 2018 Presenter: Cecilia Kretz, CDC/ New York City Public Health Laboratory, New York, NY, Phone: 323.309.1770, Email: ckretz@health.nyc.gov M. Khubbar 1 , B. Fanelli 2 , N. Hasan 2 , J. Wojnar 1 , V. Kalve 1 , R. Colwell 3 , S. Bhattacharyya 1 ; 1 Milwaukee Health Department Laboratory, Milwaukee, WI, 2 CosmosID, Inc., Rockville, MD, 3 University of Maryland, Baltimore, MD Introduction: Microbial ecology of recreational beach waters is one of the most underexplored ecosystems, often disrupted by anthropogenic activities, particularly in summer months. Here we deciphered microbial ecology of three Milwaukee beaches using state of the art whole genome shotgun metagenomics to explore microbial diversity, community resistome, virulome and their possible implication to public health. Methods: A total of 18 pooled water samples were selected based on disparity between findings by Colilert and qPCR methods. Samples were collected during summer 2016 at three Milwaukee area beaches- Bradford, McKinley and South Shore. DNA extractions of 100 ml filtrates were performed using Fecal DNA kit (Zymo Research). dsDNA quantification was performed using Qubit 4 Fluorometer. Lyophilized SmartBeads (BioGx, Inc.) were used for E. coli. Nextera XT library prep protocol was used to generate fragment libraries followed by 150 bp paired end sequencing on Illumina HiSeq4000, generating an average of 40M read pairs per sample. CosmosID bioinformatics platform was used for multi-kingdom microbiome analyses and profiling of community resistome and virulome. Results: Diverse population of bacteria (n=645), fungi (n=14), parasites (n=20), viruses and bacteriophages (n=21) are identified in all samples. Chao diversity index indicated diverse bacterial population on a given day as compared to fungi, protists and viruses. Proteobacteria (60-68%), Actinobacteria (24-27%), Bacteroidetes (4-7%), Firmicutes (0.6-1%), Verrucomicrobia (0.6-2%), Cyanobacteria (0.3-2%), Planctomycetes (0.6-0.9%) represents the dominant bacterial and Basidiomycota (36-89%) and Ascomycota (11-64%) as dominant fungal phyla detected PublicHealthLabs @APHL APHL.org