Lab Matters Fall 2023 | Page 85

APHL 2023 POSTER ABSTRACTS

PARTNERSHIPS , ACADEMIC , RESEARCH AND INDUSTRY COLLABORATIONS , TRAINING AND OUTREACH

Five Lessons Learned in Developing Training Videos for High Containment Facilities

D . Boyles , J . Bickell , S . Shearrer , S . Altmann , L . Gardiner ; MRIGlobal

Objectives : MRIGlobal ’ s biosafety team traveled to a client ’ s agricultural research facility to begin filming a series of scripted biosafety training videos . The goal of the trip was to assess and optimize the functionality of the current filming process . Methods : Prior to the visit , MRIGlobal wrote video scripts on topics including select agents , laboratory waste treatment , laboratory-acquired infections , and biosafety cabinets . Scripts were delivered to the client for feedback and approval . After approval , documents outlining specific spaces and props needed for each scene of each video were developed and submitted to the client in preparation for the filming trip . MRIGlobal then worked with a film crew to review scripts and create a filming schedule before departing for a 10-day filming trip . Discussion : During this visit , the team identified five areas unique to filming that required distinctive preparation compared to traditional training materials . We refer to these as the “ five lessons learned .” These lessons spanned the course of the development process from script writing to acting and post-production editing and can be summarized as : ( 1 ) adopt an appropriate writing style ; ( 2 ) plan scenes as you write ; ( 3 ) plan your resources ; ( 4 ) provide oversight for actors ; and ( 5 ) know your set ahead of time . These lessons are applicable to filming training videos in all settings but specifically help address some of the challenges of filming within biocontainment spaces . Conclusions : As a result of these lessons learned , a series of templates were developed that address these topics in advance to ensure that , once on site , filming can proceed without interruption . These tools created from the lessons learned can benefit anyone looking to develop training videos .

Presenter : Devin Boyles , dboyles158 @ gmail . com

Evaluation of Antiviral Properties of Amniotic Fluid-derived Extracellular Vesicles in Experimental Models of Influenza Virus Infection

C . Guenther 1 , C . Burgess 1 , M . Shipley 1 , D . Eddins 1 , A . Kumar 1 , T . del Rivero 2 , M . Bellio 2 , M . Mitrani 2 , S . Sambhara 1 , S . Gangappa 1 ;

1

US Centers for Disease Control and Prevention , 2 Organicell Regenerative Medicine

Influenza is one of the most frequently occurring viruses , with different strains circulating each year . Due to its prevalence and rapid mutation rate , it is imperative to research and develop novel antivirals that constrain influenza pathogenesis . Zofin™ is an amniotic fluid-derived therapeutic composed of naturally produced extracellular vesicles . Extracellular vesicles include both exosomes and microvesicles , which contain cargo , such as miRNA and proteins , that are used by host cells for intercellular communication and to regulate inflammation . This study explores the antiviral effects of Zofin™ using in-vitro and in-vivo models of influenza A virus ( PR8 ; H1N1 subtype ) infection . For in vitro experiments , human lung epithelial cells ( A549 ) were infected at two different multiplicities of infection ( MOI of 1 [ low ] or a MOI of 10 [ high ]) and treated with either 5 % or 25 % of Zofin™ . Cell culture supernatants were collected at 24 and 48 hours post-infection ( hpi ). For the invivo study , groups of Balb / c mice were infected , treated intranasally with Zofin™ , and lung tissue homogenates were collected at four and seven days post-infection ( dpi ). Virus titers were determined by plaque assays for PR8-infected , Zofin™-treated cells / animals and compared to PBS-treated vehicle controls . Interestingly , we demonstrate that Zofin™-treated PR8-infected cell cultures have a significant reduction in virus titer at 24 hpi at both MOIs . At 48 hpi , a significant reduction was seen for the 25 % Zofin treated infected cell cultures at both MOIs . Conversely , the antiviral effects seen in-vitro were not observed in-vivo at either 4 or 7 dpi . Ongoing studies are aimed at defining the mechanisms by which Zofin™ exerts antiviral effects in cell culture models and further research is needed to delineate any possible species-specific ( human vs mouse ) divergences in antiviral potential of Zofin™ observed in our experimental models .

Presenter : Camy Guenther , tzo9 @ cdc . gov

Experience Working on a Pilot Project with Public Health Laboratories and the CDC from a Fellow ’ s Perspective

A . Lo , D . Mallal , A . Rossheim , S . Matzinger ; Colorado Department of Public Health and Environment

Current national PulseNet surveillance protocols require culture and isolation to perform whole genome sequencing ( WGS ). Many clinical laboratories have transitioned to culture-independent diagnostic tests to identify enteric pathogens , increasing the burden on public health laboratories to perform culture and isolation in addition to WGS . To address this , US Centers for Disease Control and Prevention ( CDC ) proposed an alternative method to characterize enteric pathogens directly from stool samples via a highly multiplexed amplicon sequencing ( HMAS ) protocol . One food safety fellow was assigned to the Colorado Department of Public Health and Environment ( CDPHE ) to work on this pilot protocol during the 2022 – 2023 APHL Fellowship cycle . Overall , due to decreased cost , high throughput and likely ability to hit Salmonella HMAS targets ( i . e ., serotyping , antimicrobial resistance , virulence , core genome multilocus sequence typing ), we expect stool HMAS to be a useful option to decrease manual efforts at the state public health laboratory for characterization of Salmonella specimens for national and local cluster detection and foodborne public health response . Currently , CDPHE is one of two state public health lab pilot sites tasked with performing side-by-side comparison of the JUNO HMAS protocol with the current isolate-based PulseNet WGS protocol on Salmonella isolates and the original stool from which the isolates were cultured . This will be accomplished by gathering three pieces of data for paired isolate and stool samples : Isolatebased PulseNet WGS , isolate-based HMAS and stool-based HMAS sequencing results . Under the supervision of mentors and other technicians , the fellow learned current PulseNet WGS protocols , summating in obtaining the PulseNet WGS lab and analysis certifications . Additionally , the fellow assisted in the development and implementation of the JUNO HMAS protocol on Salmonella isolates and stool samples . This included extraction from stool and bacterial isolates , WGS and HMAS sequencing library preparation , troubleshooting of proposed protocols and communication with CDC ’ s culture-independent metagenomics subtyping group ( CIMS ) enterics group , other participating public health laboratories and