scholarly journals No Evidence of SARS-CoV-2 Among Flies or Cockroaches in COVID-19 Positive Households

2021 ◽  
Author(s):  
Christopher M Roundy ◽  
Sarah A. Hamer ◽  
Italo B. Zecca ◽  
Edward B. Davila ◽  
Lisa D. Auckland ◽  
...  
Keyword(s):  
Companion Animals ◽  
Mechanical Transmission ◽  
Viral Detection ◽  
Viral Inactivation ◽  
House Flies ◽  
Turkey Coronavirus ◽  
Insect Trap ◽  
Level 3 ◽  
Natural Settings ◽  
Biosafety Level

Abstract Background:The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in a pandemic of coronavirus disease (COVID-19), which continues to cause infections and mortality worldwide. SARS-CoV-2 is transmitted primarily via the respiratory route and has experimentally been found to be stable on surfaces for multiple days. Flies (Diptera) and other arthropods mechanically transmit several pathogens, including turkey coronavirus. A previous experimental study demonstrated house flies, Musca domestica, can mechanically transmit SARS-CoV-2, but the ability of flies in general to acquire and deposit this virus in natural settings has not been explored. The purpose of this study was to explore the possibility of mechanical transmission of SARS-CoV-2 by peridomestic insects and their potential as a xenosurveillance tool for detection of the virus.Methods:In order to optimize chances of viral detection, flies were trapped in homes where at least one confirmed human COVID-19 case(s) resided. Sticky and liquid baited fly traps were deployed inside and outside of the homes of SARS-CoV-2 human cases in Brazos, Bell, and Montgomery Counties, from June to September 2020. Flies from sticky traps were identified, pooled by taxa, homogenized, and tested for the presence of SARS-CoV-2 RNA using qRT-PCR. Liquid traps were drained, and the collected fluid similarly tested after RNA concentration. Experimental viral detection pipeline and viral inactivation were confirmed in a Biosafety Level 3 lab. As part of a separate ongoing study, companion animals in the home were sampled and tested for SARS-CoV-2 on the same day of insect trap deployment.Results:We processed the contents of 133 insect traps from 44 homes, which contained over 1,345 individual insects of 11 different Diptera families and Blattodea.These individuals were grouped into 243 pools, and all tested negative for SARS-CoV-2 RNA. Dead flies exposed to SARS-CoV-2 in a BSL3 lab were processed using the same methods and viral RNA was detected by RT-PCR. Fourteen traps in seven homes were deployed on the day that cat or dog samples tested positive for SARS-CoV-2 RNA by nasal, oral, body, or rectal samples.Conclusions:This study presents evidence that biting and non-biting flies are not likely to contribute to mechanical transmission of SARS-CoV-2 or be useful in xenosurveillance for SARS-CoV-2.

scholarly journals Network Experiences from a Cross-Sector Biosafety Level-3 Laboratory Collaboration: A Swedish Forum for Biopreparedness Diagnostics

2017 ◽  
Vol 15 (4) ◽  
pp. 384-391 ◽  
Author(s):  
Johanna Thelaus ◽  
Anna Lindberg ◽  
Susanne Thisted Lambertz ◽  
Mona Byström ◽  
Mats Forsman ◽  
...  
Keyword(s):  
Level 3 ◽  
Biosafety Level

scholarly journals Establishment of Biosafety Level 3 Laboratory Infrastructure to Study with Highly Pathogen Organisms and Principles of Operation: Review

2014 ◽  
Vol 34 (1) ◽  
pp. 120-136
Author(s):  
Fatıma YÜCEL ◽  
Hivda ÜLBEĞİ POLAT ◽  
Esin AKÇAEL ◽  
Taşkın DENİZ
Keyword(s):  
Level 3 ◽  
Biosafety Level

scholarly journals The Broad Clinical Spectrum of Disseminated Histoplasmosis in HIV-Infected Patients: A 30 Years’ Experience in French Guiana

2019 ◽  
Vol 5 (4) ◽  
pp. 115 ◽  
Author(s):  
Pierre Couppié ◽  
Katarina Herceg ◽  
Morgane Bourne-Watrin ◽  
Vincent Thomas ◽  
Denis Blanchet ◽  
...  
Keyword(s):  
French Guiana ◽  
Histoplasma Capsulatum ◽  
Opportunistic Infections ◽  
Clinical Findings ◽  
Diagnostic Methods ◽  
Medical Mycology ◽  
Time To Initiation ◽  
Level 3 ◽  
Endemic Areas ◽  
Biosafety Level

Histoplasmosis is a common but neglected AIDS-defining condition in endemic areas for Histoplasma capsulatum. At the advanced stage of HIV infection, the broad spectrum of clinical features may mimic other frequent opportunistic infections such as tuberculosis and makes it difficult for clinicians to diagnose histoplasmosis in a timely manner. Diagnosis of histoplasmosis is difficult and relies on a high index of clinical suspicion along with access to medical mycology facilities with the capacity to implement conventional diagnostic methods (direct examination and culture) in a biosafety level 3 laboratory as well as indirect diagnostic methods (molecular biology, antibody, and antigen detection tools in tissue and body fluids). Time to initiation of effective antifungals has an impact on the patient’s prognosis. The initiation of empirical antifungal treatment should be considered in endemic areas for Histoplasma capsulatum and HIV. Here, we report on 30 years of experience in managing HIV-associated histoplasmosis based on a synthesis of clinical findings in French Guiana with considerations regarding the difficulties in determining its differential diagnosis with other opportunistic infections.

scholarly journals Propagation, Inactivation, and Safety Testing of SARS-CoV-2

Viruses ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 622 ◽  
Author(s):  
Alexander S. Jureka ◽  
Jesus A. Silvas ◽  
Christopher F. Basler
Keyword(s):  
Plaque Assay ◽  
Virus Infectivity ◽  
Safety Testing ◽  
Level 3 ◽  
Chinese Province ◽  
Infected Cells ◽  
Multiple Cell ◽  
Novel Coronavirus ◽  
Effective Procedures ◽  
Biosafety Level

In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly described and effective procedures that enable the propagation and quantification of infectious virus. As work with the virus is recommended to be performed at biosafety level 3, validated methods to effectively inactivate the virus to enable the safe study of RNA, DNA, and protein from infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat.

Achieving Biosafety Level 3 through the Use of Biosafety Level 2 Facilities and Biosafety Level 3 Practices: A Prevalence Survey of Medical Research and Academic Institutions

1997 ◽  
Vol 2 (4) ◽  
pp. 43-46 ◽  
Author(s):  
Robert J. Emery ◽  
Pek Lee ◽  
James Garman
Keyword(s):  
Research Work ◽  
Health And Safety ◽  
Mail Survey ◽  
Prevalence Survey ◽  
Strict Adherence ◽  
Level 3 ◽  
Level 2 ◽  
Safety Guidelines ◽  
Biosafety Level ◽  
Biosafety Level 2

Heightened interest in pathogens with the potential for aerosol transmission and for which prevention and medical treatment is not readily available has resulted in a need for more work environments that meet Biosafety Level 3 (BSL 3) criteria. Recognizing that the facility-based criteria for BSL 3 cannot be achieved by some existing laboratories, the Centers for Disease Control and Prevention (CDC) and National Institutes of Health (NIH) biological safety guidelines provide an option for attaining BSL 3 status through the use of Biosafety Level 2 (BSL 2) facilities and strict adherence to BSL 3 practices (BSL 2/3). Inherent to this provision is a greater emphasis on safe work practices. Since the extent to which this approach is actually used in practice is not known, a nationwide mail survey of medical academic and research institutions was conducted to provide an objective indication of the proportion of BSL 3 operations actually being carried out in the BSL 2/3 mode. The results obtained indicate that 2% of activities designated as BSL 3 in the study population actually achieve this level of protection using the BSL 2/3 approach. The findings quantitatively estimate for the first time the proportion of BSL 3 activities being carried out in this fashion, and can serve as a reference point for future studies to evaluate usage trends. The results also demonstrate the utility of flexible, performance-based health and safety guidelines, as a significant amount of clinical and research work is being accommodated with the BSL 2/3 provision.

Research Activities of Hokudai Center for Zoonosis Control in Zambia

2014 ◽  
Vol 9 (5) ◽  
pp. 818-822
Author(s):  
Hideaki Higashi ◽  
◽  
Hiroshi Kida
Keyword(s):  
South Africa ◽  
Infectious Diseases ◽  
Emerging Infectious Diseases ◽  
Epidemiological Studies ◽  
Zoonotic Diseases ◽  
University Research ◽  
Research Activities ◽  
Level 3 ◽  
The University ◽  
Biosafety Level

The Hokkaido University Research Center for Zoonosis Control (CZC) established the Hokudai Center for Zoonosis Control in Zambia (HUCZCZ) at the School of Veterinary Medicine, the University of Zambia, in 2007 to control zoonotic diseases in the areas of South Africa, where various emerging infectious diseases have occurred. The CZC promotes epidemiological studies and basic researches of infectious diseases caused by viruses, protozoa, and bacteria by using the biosafety level 3 facility in the HUCZCZ. This article introduces research activities of the HUCZCZ in Zambia.

Certification of Biosafety Level 3 (BSL3) Facilities

1996 ◽  
Vol 1 (1) ◽  
pp. 26-51 ◽  
Author(s):  
Esmeralda Party ◽  
James Reiman ◽  
Edward L. Gershey
Keyword(s):  
Environmental Protection ◽  
Review Process ◽  
Infectious Agents ◽  
Level 3 ◽  
Work Done ◽  
Biosafety Level

The increasing need to work with airborne infectious agents places greater reliance upon the personnel and environmental protection that biocontainment facilities provide. Ensuring that these facilities function properly depends on carefully reviewing post-construction performance and establishing operating protocols that take advantage of a facility's attributes while recognizing its individual limitations. Our review process expands on the work done by others and provides a foundation for developing standard certification procedures for biocontainment facilities.

scholarly journals A Universal Next-Generation Sequencing Protocol To Generate Noninfectious Barcoded cDNA Libraries from High-Containment RNA Viruses

mSystems ◽  
2016 ◽  
Vol 1 (3) ◽  
Author(s):  
Lindsey A. Moser ◽  
Lisbeth Ramirez-Carvajal ◽  
Vinita Puri ◽  
Steven J. Pauszek ◽  
Krystal Matthews ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Operating Procedure ◽  
Rna Viruses ◽  
Standard Operating Procedure ◽  
Next Generation ◽  
Standard Operating ◽  
Level 3 ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing ◽  
Biosafety Level

ABSTRACT This report establishes and validates a standard operating procedure (SOP) for select agents (SAs) and other biosafety level 3 and/or 4 (BSL-3/4) RNA viruses to rapidly generate noninfectious, barcoded cDNA amenable for next-generation sequencing (NGS). This eliminates the burden of testing all processed samples derived from high-consequence pathogens prior to transfer from high-containment laboratories to lower-containment facilities for sequencing. Our established protocol can be scaled up for high-throughput sequencing of hundreds of samples simultaneously, which can dramatically reduce the cost and effort required for NGS library construction. NGS data from this SOP can provide complete genome coverage from viral stocks and can also detect virus-specific reads from limited starting material. Our data suggest that the procedure can be implemented and easily validated by institutional biosafety committees across research laboratories. Several biosafety level 3 and/or 4 (BSL-3/4) pathogens are high-consequence, single-stranded RNA viruses, and their genomes, when introduced into permissive cells, are infectious. Moreover, many of these viruses are select agents (SAs), and their genomes are also considered SAs. For this reason, cDNAs and/or their derivatives must be tested to ensure the absence of infectious virus and/or viral RNA before transfer out of the BSL-3/4 and/or SA laboratory. This tremendously limits the capacity to conduct viral genomic research, particularly the application of next-generation sequencing (NGS). Here, we present a sequence-independent method to rapidly amplify viral genomic RNA while simultaneously abolishing both viral and genomic RNA infectivity across multiple single-stranded positive-sense RNA (ssRNA+) virus families. The process generates barcoded DNA amplicons that range in length from 300 to 1,000 bp, which cannot be used to rescue a virus and are stable to transport at room temperature. Our barcoding approach allows for up to 288 barcoded samples to be pooled into a single library and run across various NGS platforms without potential reconstitution of the viral genome. Our data demonstrate that this approach provides full-length genomic sequence information not only from high-titer virion preparations but it can also recover specific viral sequence from samples with limited starting material in the background of cellular RNA, and it can be used to identify pathogens from unknown samples. In summary, we describe a rapid, universal standard operating procedure that generates high-quality NGS libraries free of infectious virus and infectious viral RNA. IMPORTANCE This report establishes and validates a standard operating procedure (SOP) for select agents (SAs) and other biosafety level 3 and/or 4 (BSL-3/4) RNA viruses to rapidly generate noninfectious, barcoded cDNA amenable for next-generation sequencing (NGS). This eliminates the burden of testing all processed samples derived from high-consequence pathogens prior to transfer from high-containment laboratories to lower-containment facilities for sequencing. Our established protocol can be scaled up for high-throughput sequencing of hundreds of samples simultaneously, which can dramatically reduce the cost and effort required for NGS library construction. NGS data from this SOP can provide complete genome coverage from viral stocks and can also detect virus-specific reads from limited starting material. Our data suggest that the procedure can be implemented and easily validated by institutional biosafety committees across research laboratories.

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