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:
[email protected] 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:
[email protected]
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
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