APHL 2022 POSTER ABSTRACTS morbidity areas of the state of Wisconsin will include support as a reference lab and SURRG – Center of Excellence ( COE ) to perform N . gonorrhoeae-Antimicrobial Susceptibility Testing ( GC-AST ) for patients with suspect treatment failure , and for culture confirmed cases of gonococcal disseminated infection ( DGI ).
Methods : N . gonorrhoeae is identified from InTrays with oxidase positive colonies , gram-negative diplococci and confirmed by api NH . AST is performed using Etest with Ciprofloxacin ( CI ), Ceftriaxone ( TX ), Cefixime ( IX ) and Gentamicin ( GM ) gradient strips . Surveillance for ARGC from genital , rectal and pharyngeal sources conducted at six designated SURRG clinics – STD clinics and five communitybased clinics in Milwaukee . Isolates are sent to Tennessee Antibiotic Resistance Lab Network ( ARLN ) for confirmatory AST using agar dilution . Isolates from patients reported with DGI are sent to CDC for genomic characterization .
Results : A total of 390 patients ( 445 isolates / 2779 positive cultures ), were identified with ARGC . Majority of ARGC isolates ( 440 ; 98.9 %) isolates had reduced susceptibility ( RS ) to Azithromycin . During five years of SURRG Cycle 1 , very few were identified with RS to cephalosporins ( 2 Ceftriaxone ; 3 Cefixime ). No treatment failures have been observed . During CY2019 – CY2021 , 29 cases of culture confirmed DGI were reported in Wisconsin . Of the 17 isolates shared with MHDL for GC-AST , with culture confirmed DGI , two isolates had RS to Azithromycin . None of the DGI isolates were resistant to cephalosporins .
Conclusions : The Southeastern Region of Wisconsin has the highest reported rate of GC in the state ( 183 per 100,000 in CY 2020 ). MHDL serves a critical role in this high morbidity region as it continues to provide GC-AST surveillance activities employing rapid methods to detect ARGC in patients in diverse set of select SURRG clinics with high rates of reported GC morbidity . As MHDL expands its capacity as a SURRG-COE reference laboratory during SURRG Cycle 2 , these PHL services will be an important statewide resource to support GC-AST for patients with suspect treatment failure and patients with culture confirmed DGI . While genomic analyses of DGI isolates at CDC help understanding and characterization of disseminated gonococcal infections , building local cultureindependent and genomic capacity will allow timely addressing increasing trends of reported DGI observed in Wisconsin and across the country .
Presenter : Suzanna Budiono , City of Milwaukee Health Department , sbudio @ milwaukee . gov
Comparison of Environmental and Clinical isolates of Legionella pneumophila Associated with a Long-Term Care Facility using Next Generation Sequencing
S Nguyen , J Doss , R Blackwell , S Merid , B Hamilton , D Edwards , E Vaughn , K Tran and J Hauser , DC Department of Forensic Sciences
Legionella pneumophila is an opportunistic pathogen that infects amoebas in freshwater environments . The ability to persist in these protozoan hosts lends it the ability to survive as intracellular parasites in humans . Clinical diseases associated with Legionella species are often found in immunocompromised patients such as neonates and the elderly . Genomic surveillance of L . pneumophila is limited in comparison to well established pathogen surveillance programs such as foodborne pathogens with PulseNet . Here , we detail the use of next generation sequencing methods to identify genetic links between an environmental isolate of L . pneumophila with a clinical isolate collected from a long-term care facility in the District of Columbia and a resident with confirmed legionellosis in 2021 . The long-term care facility has had a history of Legionella infections and additional isolates collected in 2018 were also compared . Sequence-based typing ( SBT ) of these isolates identified the Legionella as ST224 , a member of L . pneumophila serogroup 1 . Additionally , the use of a Single Nucleotide Polymorphisms Pipeline ( SNP ) analysis showed the environmental isolate and the clinical isolate had no SNP differences , indicating an environmental source for legionellosis cases at this facility . Our study demonstrates the use of next generation sequencing analysis to determine relatedness among environmental and clinical Legionella isolates .
Presenter : Scott Nguyen , DC Department of Forensic Sciences , scott . nguyen @ dc . gov
Improved Genomic Characterization of Clinical Bacterial Isolates of Unknown Identification in Florida using Hybrid Assemblies from Short- and Long-read Sequencing Platforms
T Bazile 1 , 2 , M Hogshead 1 , J Blanton 1 , S Schmedes 1 ; 1 Florida Bureau of Public Health Laboratories , 2 Association of Public Health Laboratories
The Microbiology and Molecular Departments at the Florida Bureau of Public Health Laboratories ( BPHL ) regularly receive clinical bacterial specimens from unknown etiologies as culture for identification ( CFI ). Whole-genome sequencing is readily performed for genetic identification and characterization of the bacterial specimens that traditional culture and biochemical methods cannot identify . BPHL has sequenced and assembled over 20 draft genomes from CFI specimens using short-read sequencing data , generated on Illumina platforms , that continue to remain difficult to identify . This is likely due to minimal representation at the genus or species level in public genomic repositories . To improve the genomic characterization of CFIs and to increase the representation in public genomic databases , long-read sequencing was used to improve the draft assemblies . Hybrid assemblies were generated for each CFI using short-read and long-read sequencing data , generated from the Oxford Nanopore Technologies minION , to improve characterization of these minimally represented organisms . With both the accessibility to more complete regions of the genome and the continuing progress in accuracy , long-read sequencing has the potential to enable improved CFI profiling to aid in the identification of hard-to-identify clinical bacterial specimens . Improved genetic characterization and identification of these specimens facilitates more effective public health responses to the related clinical cases .
Presenter : Tassy Bazile , Florida Bureau of Public Health Laboratories , Tassy . Bazile @ flhealth . gov
Infectious Diseases