The efficacy of medical masks and respirators against respiratory infection in healthcare workers

Objective: We aimed to examine the efficacy of medical masks and respirators in protecting against respiratory infections using pooled data from two homogenous randomised control clinical trials (RCTs).
Methods: The data collected on 3591 subjects in two similar RCTs conducted in Beijing, China, which examined the same infection outcomes, were pooled. Four interventions were compared: (i) continuous N95 respirator use, (ii) targeted N95 respirator use, (iii) medical mask use and (iv) control arm. The outcomes were laboratory‐confirmed viral respiratory infection, influenza A or B, laboratory‐confirmed bacterial colonisation and pathogens grouped by mode of transmission.
Results: Rates of all outcomes were consistently lower in the continuous N95 and/or targeted N95 arms. In adjusted analysis, rates of laboratory‐confirmed bacterial colonisation (RR 0.33, 95% CI 0.21‐0.51), laboratory‐confirmed viral infections (RR 0.46, 95% CI 0.23‐0.91) and droplet‐transmitted infections (RR 0.26, 95% CI 0.16‐0.42) were significantly lower in the continuous N95 arm. Laboratory‐confirmed influenza was also lowest in the continuous N95 arm (RR 0.34, 95% CI 0.10‐1.11), but the difference was not statistically significant. Rates of laboratory‐confirmed bacterial colonisation (RR 0.54, 95% CI 0.33‐0.87) and droplet‐transmitted infections (RR 0.43, 95% CI 0.25‐0.72) were also lower in the targeted N95 arm, but not in medical mask arm.
Conclusion: The results suggest that the classification of infections into droplet versus airborne transmission is an oversimplification. Most guidelines recommend masks for infections spread by droplets. N95 respirators, as “airborne precautions,” provide superior protection for droplet‐transmitted infections. To ensure the occupational health and safety of healthcare worker, the superiority of respirators in preventing respiratory infections should be reflected in infection control guidelines.
REFERENCES: MacIntyre, Chandini Raina et al. “The Efficacy of Medical Masks and Respirators against Respiratory Infection in Healthcare Workers.” Influenza and Other Respiratory Viruses 11.6 (2017): 511–517.

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Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented—one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

REFERENCE:
Bowey-Dellinger, Kristen et al. “Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory.” Journal of Microbiology & Biology Education 18.2 (2017): 18.2.38. PMC. Web. 4 Jan. 2018.


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Biosafety and Biosecurity in European Containment Level 3 Laboratories: Focus on French Recent Progress and Essential Requirements

Even if European Union (EU) Member States are obliged to implement EU Directives 2000/54/EC on the protection of workers from risks related to exposure to biological agents at work, national biosafety regulations and practices varied from country to country. In fact, EU legislation on biological agents and genetically modified microorganisms is often not specific enough to ensure harmonization leading to difficulties in implementation for most laboratories. In the same way, biosecurity is a relatively new concept and a few EU Member States are known to have introduced national laboratory biosecurity legislation. In France, recent regulations have reinforced biosafety/biosecurity in containment level 3 (CL-3) laboratories but they concern a specific list of pathogens with no correlation in other European Members States. The objective of this review was to summarize European biosafety/biosecurity measures concerning CL-3 facilities focusing on French specificities. Essential requirements needed to preserve efficient biosafety measures when manipulating risk group 3 biological agents are highlighted. In addition, International, European and French standards related to containment laboratory planning, operation or biosafety equipment are described to clarify optimal biosafety and biosecurity requirements.

REFERENCE:
Pastorino, Boris, Xavier de Lamballerie, and Rémi Charrel. “Biosafety and Biosecurity in European Containment Level 3 Laboratories: Focus on French Recent Progress and Essential Requirements.” Frontiers in Public Health 5 (2017): 121.


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Basic Scholarship in Biosafety Is Critically Needed To Reduce Risk of Laboratory Accidents

Our firm conducted a risk/benefit assessment of “gain-of-function” research, as part of the deliberative process following a U.S. moratorium on the research (U.S. Department of Health and Human Services, U.S. Government Gain-of-Function Deliberative Process and Research Funding Pause on Selected Gain-of-Function Research Involving Influenza, MERS, and SARS Viruses, 2014). Due to significant missing but theoretically acquirable data, our biosafety assessment faced limitations, and we were forced to provide a relative, instead of absolute, measure of risk (Gryphon Scientific, LLC, Risk and Benefit Analysis of Gain of Function Research, 2016). Here, we argue that many of these types of missing data represent large and stunning gaps in our knowledge of biosafety and argue that these missing data, once acquired via primary research efforts, would improve biosafety risk assessments and could be incorporated into biosafety practices to reduce risk of accidents. Governments invest billions in biological research; at least a small fraction of this support is warranted to prevent biological accidents.
REFERENCE:
Ritterson, Ryan, and Rocco Casagrande. “Basic Scholarship in Biosafety Is Critically Needed To Reduce Risk of Laboratory Accidents.” Ed. Michael J. Imperiale. mSphere 2.2 (2017): e00010–17.


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Biosafety Level 3 setup for multiphoton microscopy in vivo

Multiphoton microscopy has revealed important insights into cellular behavior in vivo. However, its application in infectious settings often encounters technical, safety and regulatory limitations that prevent its wider use with highly virulent human pathogens. Herein, we present a method that renders multiphoton microscopy in vivo compatible with biosafety level 3 regulations and present an example of its application and potential to visualize a Mycobacterium tuberculosis infection of the mouse lung.
REFERENCE:
Barlerin, D. et al. “Biosafety Level 3 Setup for Multiphoton Microscopy in Vivo.” Scientific Reports 7 (2017): 571. PMC. Web. 4 Jan. 2018.


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Non-occupational Uses of Respiratory Protection

A respirator is one method for reducing personal exposure to particulate respiratory hazards. NIOSH has been certifying respirators and conducting research on these devices for over 40 years. Along with our collaborators in government, industry, and academia, we have accumulated a wealth of knowledge on how respirators perform in workplace settings. In the last decade, respirator use by the general public has become a more frequent topic of debate as public health officials at the local, state, and federal level consider which public health and non-pharmaceutical interventions to recommend. In this blog, we have teamed with colleagues from across the Centers for Disease Control and Prevention (CDC) to assist state and federal public health agencies, and other safety professionals, by translating the lessons we’ve learned from respirator use at work to respirator use by the general public.
REFERENCE:
CDC. Non-occupational Uses of Respiratory Protection – What Public Health Organizations and Users Need to Know. By Ronald Shaffer et al.
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The ghosts of HeLa: How cell line misidentification contaminates the scientific literature

While problems with cell line misidentification have been known for decades, an unknown number of published papers remains in circulation reporting on the wrong cells without warning or correction. Here we attempt to make a conservative estimate of this ‘contaminated’ literature. We found 32,755 articles reporting on research with misidentified cells, in turn cited by an estimated half a million other papers. The contamination of the literature is not decreasing over time and is anything but restricted to countries in the periphery of global science. The decades-old and often contentious attempts to stop misidentification of cell lines have proven to be insufficient. The contamination of the literature calls for a fair and reasonable notification system, warning users and readers to interpret these papers with appropriate care.

REFERENCE:
Horbach SPJM, Halffman W (2017) The ghosts of HeLa: How cell line misidentification contaminates the scientific literature. PLoS ONE 12(10): e0186281. https://doi.org/10.1371/journal.pone.0186281

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Assessing the Biological Safety Profession's Evaluation and Control of Risks... #FieldSampling

FRAGMENT: This study developed a web-based survey distributed to practicing biological safety professionals to determine the prevalence of and extent to which biological safety programs consider and evaluate field collection activities. In cases where such issues were considered, the data collected characterize the types of controls and methods of oversight at the institutional level that are employed. Sixty-one percent (61%) of the survey respondents indicated that research involving the field collection of biological specimens is conducted at their institutions. A majority (79%) of these field collection activities occur at academic institutions. Twenty-seven percent (27%) of respondents indicated that their safety committees do not consider issues related to biological specimens collected in the field, and only 25% with an oversight committee charged to review field collection protocols have generated a field research-specific risk assessment form to facilitate the assembly of pertinent information for a project risk assessment review. The results also indicated that most biosafety professionals (73% overall; 71% from institutions conducting field collection activities) have not been formally trained on the topic, but many (64% overall; 87% from institutions conducting field collection activities) indicated that training on field research safety issues would be helpful, and even more (71% overall; 93% from institutions conducting field collection activities) would consider participation in such a training course. Results obtained from this study can be used to develop a field research safety toolkit and associated training curricula specifically targeted to biological safety professionals.
REFERENCE:
Patlovich ST, et al. Assessing the Biological Safety Profession's Evaluation and Control of Risks Associated with the Field Collection of Potentially Infectious Specimens. Appl Biosaf. 2015 Mar; 20(1): 27–40. Author manuscript; available in PMC 2018 Jan 9.

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Safe-by-Design: from Safety to Responsibility

Safe-by-design (SbD) aims at addressing safety issues already during the R&D and design phases of new technologies. SbD has increasingly become popular in the last few years for addressing the risks of emerging technologies like nanotechnology and synthetic biology. We ask to what extent SbD approaches can deal with uncertainty, in particular with indeterminacy, i.e., the fact that the actual safety of a technology depends on the behavior of actors in the value chain like users and operators. We argue that while indeterminacy may be approached by designing out users as much as possible in attaining safety, this is often not a good strategy. It will not only make it more difficult to deal with unexpected risks; it also misses out on the resources that users (and others) can bring for achieving safety, and it is undemocratic. We argue that rather than directly designing for safety, it is better to design for the responsibility for safety, i.e., designers should think where the responsibility for safety is best situated and design technologies accordingly. We propose some heuristics that can be used in deciding how to share and distribute responsibility for safety through design.
REFERENCE:
Van de Poel, Ibo, and Zoë Robaey. “Safe-by-Design: From Safety to Responsibility.” Nanoethics 11.3 (2017): 297–306. PMC. Web. 8 Jan. 2018.

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National Framework for Personal Protective Equipment Conformity Assessment - Infrastructure

The goal of our efforts at the National Institute for Occupational Safety and Health (NIOSH) is to provide national and world leadership to prevent workplace illnesses and injuries. We accomplish this by conducting and supporting activities to protect workers from work-related exposures to hazards. One core objective of this approach involves the development and use of personal protective equipment (PPE). Workers are more likely to appropriately use PPE when they are confident that the equipment will provide the intended protections based on its conformance with appropriate standards. The National Academies of Sciences, Engineering, and Medicine (the Academies) indicates that “for the consumer or worker, conformity assessment provides confidence in the claims made about the product by the manufacturer and may assist the consumer with purchasing decisions in determining the fitness of a product for it its intended use.” [IOM, 2011, page 3] A comprehensive and tailor-made conformity assessment (CA) program is the most effective way to manage risks of a non-conforming PPE and instill this confidence in PPE users.
REFERENCE:
NIOSH [2017]. National framework for personal protective equipment conformity assessment – infrastructure. By D’Alessandro M. Pittsburgh, PA: U.S. Department of Health and Human Services, Centers for Disease Control and Prevention, National Institute for Occupational Safety and Health, DHHS (NIOSH) Publication 2018–102.

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CDC Safety Training Course for Ebola Virus Disease Healthcare Workers

Response to sudden epidemic infectious disease emergencies can demand intensive and specialized training, as demonstrated in 2014 when Ebola virus disease (EVD) rapidly spread throughout West Africa. The medical community quickly became overwhelmed because of limited staff, supplies, and Ebola treatment units (ETUs). Because a mechanism to rapidly increase trained healthcare workers was needed, the US Centers for Disease Control and Prevention developed and implemented an introductory EVD safety training course to prepare US healthcare workers to work in West Africa ETUs. The goal was to teach principles and practices of safely providing patient care and was delivered through lectures, small-group breakout sessions, and practical exercises. During September 2014–March 2015, a total of 570 participants were trained during 16 course sessions. This course quickly increased the number of clinicians who could provide care in West Africa ETUs, showing the feasibility of rapidly developing and implementing training in response to a public health emergency.
REFERENCE:
Narra, Rupa et al. “CDC Safety Training Course for Ebola Virus Disease Healthcare Workers.” Emerging Infectious Diseases 23.Suppl 1 (2017): S217–S224.

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Network Experiences from a Cross-Sector Biosafety Level-3 Laboratory Collaboration

The Swedish Forum for Biopreparedness Diagnostics (FBD) is a network that fosters collaboration among the 4 agencies with responsibility for the laboratory diagnostics of high-consequence pathogens, covering animal health and feed safety, food safety, public health and biodefense, and security. The aim of the network is to strengthen capabilities and capacities for diagnostics at the national biosafety level-3 (BSL-3) laboratories to improve Sweden's biopreparedness, in line with recommendations from the EU and WHO. Since forming in 2007, the FBD network has contributed to the harmonization of diagnostic methods, equipment, quality assurance protocols, and biosafety practices among the national BSL-3 laboratories. Lessons learned from the network include: (1) conducting joint projects with activities such as method development and validation, ring trials, exercises, and audits has helped to build trust and improve communication among participating agencies; (2) rotating the presidency of the network steering committee has fostered trust and commitment from all agencies involved; and (3) planning for the implementation of project outcomes is important to maintain gained competencies in the agencies over time. Contacts have now been established with national agencies of the other Nordic countries, with an aim to expanding the collaboration, broadening the network, finding synergies in new areas, strengthening the ability to share resources, and consolidating long-term financing in the context of harmonized European biopreparedness.

REFERENCE:
Thelaus, Johanna et al. “Network Experiences from a Cross-Sector Biosafety Level-3 Laboratory Collaboration: A Swedish Forum for Biopreparedness Diagnostics.” Health Security 15.4 (2017): 384–391.

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A biosafety level 2 virology lab for biotechnology undergraduates

Medical, industrial, and basic research relies heavily on the use of viruses and vectors. Therefore, it is important that bioscience undergraduates learn the practicalities of handling viruses. Teaching practical virology in a student laboratory setup presents safety challenges, however. The aim of this article is to describe the design and implementation of a virology laboratory, with emphasis on student safety, for biotechnology undergraduates. Cell culture techniques, animal virus infection, quantification, and identification are taught at a biosafety level 2 for a diverse group of undergraduates ranging from 20 to 50 students per group.

REFERENCES:
Matza‐Porges, Sigal, and Dafna Nathan. “A Biosafety Level 2 Virology Lab for Biotechnology Undergraduates.” Biochemistry and Molecular Biology Education 45.6 (2017): 537–543. PMC.

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Feliz 2018

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NOM-005-SCT/2008, Información de emergencia para el transporte de substancias, materiales y residuos peligrosos

La presente Norma Oficial Mexicana establece en forma uniforme para su aplicación en los diversos modos de transporte, los datos y especificaciones que debe contener la Información de Emergencia para el Transporte de Substancias, Materiales y Residuos Peligrosos, que indique las acciones a seguir para casos de incidentes o accidentes (fugas, derrames, explosiones, incendios, exposiciones, etc.), que debe llevar toda unidad durante el transporte de substancias, materiales y residuos peligrosos, en un lugar accesible de la unidad y retirada de la carga.

REFERENCIA:
NOM-005-SCT/2008, Información de emergencia para el transporte de substancias, materiales y residuos peligrosos.

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NOM-003-SEGOB-2011, Señales y avisos para protección civil.- Colores, formas y símbolos a utilizar.

La experiencia indica que la correcta aplicación de esta Norma Oficial Mexicana, contribuye a mejorar las condiciones de seguridad en instalaciones y sitios en los que, conforme a leyes, reglamentos y normatividad aplicable en materia de prevención de riesgos, debe implementarse un sistema de señalización sobre protección civil, en beneficio de la población que concurre o labora en ellos.

REFERENCIA:
NORMA Oficial Mexicana NOM-003-SEGOB-2011, Señales y avisos para protección civil.- Colores, formas y símbolos a utilizar.

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Lessons to be Learned from #Biosafety Incidents in the USA

During recent months, the Centers for Disease Control and Prevention (CDC) announced the occurrence of three major biosafety incidents, raising serious concern about biosafety and biosecurity guideline implementation in the most prestigious agencies in the United States: the CDC, the National Institutes of Health (NIH) and the Federal Drug Administration (FDA). These lapses included: a) the mishandling of Bacillus anthracis spores potentially exposing dozens of employees to anthrax; b) the shipment of low pathogenic influenza virus unknowingly cross-contaminated with a highly pathogenic strain; and c) an inventory lapse of hundreds of samples of biological agents, including six vials of variola virus kept in a cold storage room for decades, unnoticed. In this review we present the published data on these events, report the CDC inquiry's main findings, and discuss the key lessons to be learnt to ensure safer scientific practice in biomedical and microbiological service and research laboratories.

REFERENCE:
Weiss S, Yitzhaki S, Shapira SC. Lessons to be Learned from Recent Biosafety Incidents in the United States. Isr Med Assoc J. 2015 May;17(5):269-73. Review. PubMed PMID: 26137650.

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Enhancing Surveillance and Diagnostics in Anthrax-Endemic Countries

Naturally occurring anthrax disproportionately affects the health and economic welfare of poor, rural communities in anthrax-endemic countries. However, many of these countries have limited anthrax prevention and control programs. Effective prevention of anthrax outbreaks among humans is accomplished through routine livestock vaccination programs and prompt response to animal outbreaks. The Centers for Disease Control and Prevention uses a 2-phase framework when providing technical assistance to partners in anthrax-endemic countries. The first phase assesses and identifies areas for improvement in existing human and animal surveillance, laboratory diagnostics, and outbreak response. The second phase provides steps to implement improvements to these areas. We describe examples of implementing this framework in anthrax-endemic countries. These activities are at varying stages of completion; however, the public health impact of these initiatives has been encouraging. The anthrax framework can be extended to other zoonotic diseases to build on these efforts, improve human and animal health, and enhance global health security.

REFERENCE:

Vieira, Antonio R. et al. “Enhancing Surveillance and Diagnostics in Anthrax-Endemic Countries.” Emerging Infectious Diseases 23.Suppl 1 (2017): S147–S153. PMC. Web. 9 Dec. 2017.

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Shelf-Life of Chlorine Solutions Recommended in Ebola Virus Disease Response

In Ebola Virus Disease (EVD) outbreaks, it is widely recommended to wash living things (handwashing) with 0.05% (500 mg/L) chlorine solution and non-living things (surfaces, personal protective equipment, dead bodies) with 0.5% (5,000 mg/L) chlorine solution. Chlorine solutions used in EVD response are primarily made from powdered calcium hypochlorite (HTH), granular sodium dichloroisocyanurate (NaDCC), and liquid sodium hypochlorite (NaOCl), and have a pH range of 5–11. Chlorine solutions degrade following a reaction highly dependent on, and unusually sensitive to, pH, temperature, and concentration. We determined the shelf-life of 0.05% and 0.5% chlorine solutions used in EVD response, including HTH, NaDCC, stabilized NaOCl, generated NaOCl, and neutralized NaOCl solutions. Solutions were stored for 30 days at 25, 30, and 35°C, and tested daily for chlorine concentration and pH. Maximum shelf-life was defined as days until initial concentration fell to <90% of initial concentration in ideal laboratory conditions. At 25–35°C, neutralized-NaOCl solutions (pH = 7) had a maximum shelf-life of a few hours, NaDCC solutions (pH = 6) 2 days, generated NaOCl solutions (pH = 9) 6 days, and HTH and stabilized NaOCl solutions (pH 9–11) >30 days. Models were developed for solutions with maximum shelf-lives between 1–30 days. Extrapolating to 40°C, the maximum predicted shelf-life for 0.05% and 0.5% NaDCC solutions were 0.38 and 0.82 hours, respectively; predicted shelf-life for 0.05% and 0.5% generated NaOCl solutions were >30 and 5.4 days, respectively. Each chlorine solution type offers advantages and disadvantages to responders, as: NaDCC is an easy-to-import high-concentration effervescent powder; HTH is similar, but forms a precipitate that may clog pipes; and, NaOCl solutions can be made locally, but are difficult to transport. We recommend responders chose the most appropriate source chlorine compound for their use, and ensure solutions are stored at appropriate temperatures and used or replaced before expiring.
REFERENCIA:
Iqbal, Qais et al. “Shelf-Life of Chlorine Solutions Recommended in Ebola Virus Disease Response.” Ed. Vincent Jacobus Munster. PLoS ONE 11.5 (2016): e0156136.


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Enterococcus hirae biofilm formation on hospital material surfaces and effect of new biocides

BACKGROUND: Nowadays, the bacterial contamination in the hospital environment is of particular concern because the hospital-acquired infections (HAIs), also known as nosocomial infections, are responsible for significant morbidity and mortality. This work evaluated the capability of Enterococcus hirae to form biofilm on different surfaces and the action of two biocides on the produced biofilms.
METHODS: The biofilm formation of E. hirae ATCC 10541 was studied on polystyrene and stainless steel surfaces through the biomass quantification and the cell viability at 20 and 37 °C. The effect of LHIDROXI FAST and LH ENZYCLEAN SPRAY biocides on biomasses was expressed as percentage of biofilm reduction. E. hirae at 20 and 37 °C produced more biofilm on the stainless steel in respect to the polystyrene surface. The amount of viable cells was greater at 20 °C than with 37 °C on the two analyzed surfaces. Biocides revealed a good anti-biofilm activity with the most effect for LH ENZYCLEAN SPRAY on polystyrene and stainless steel at 37 °C with a maximum biofilm reduction of 85.72 and 86.37%, respectively.
RESULTS: E. hirae is a moderate biofilm producer depending on surface material and temperature, and the analyzed biocides express a remarkable antibiofilm action.
CONCLUSION: The capability of E. hirae to form biofilm can be associated with its increasing incidence in hospital-acquired infections, and the adoption of suitable disinfectants is strongly recommended.
REFERENCIA:
Di Lodovico S, et al. Enterococcus hirae biofilm formation on hospital material surfaces and effect of new biocides. Environ Health Prev Med. 2017 Aug 2;22(1):63. doi: 10.1186/s12199-017-0670-3. PubMed PMID: 29165147; PubMed Central PMCID: PMC5664585.

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Guidance on regulations for the transport of infectious substances 2017–2018

This publication provides information for identifying, classifying, marking, labelling, packaging, documenting and refrigerating infectious substances for transportation and ensuring their safe delivery.

The document provides practical guidance to facilitate compliance with applicable international regulations for the transport of infectious substances by all modes of transport, both nationally and internationally, and include the changes that apply from 1 January 2017. The current revision replaces the document issued by the World Health Organization (WHO) in 2015 (document WHO/HSE/GCR/2015.2). This publication, however, does not replace national and international transport regulations.

Applicable as from 1 January 2017

Authors:
World Health Organization

Publication details

Number of pages: 40
Publication date: 2017
Languages: English
WHO reference number: WHO/WHE/CPI/2017.8

Downloads

• ==> English pdf 1.63 MB <== 

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Biological samples transportation by drones: ready for prime time?

FRAGMENT:
According to the concept originally introduced by George D. Lundberg in the 1980s, the total testing process entails three essential and sequential parts, that are the preanalytical phase, the analytical phase and the postanalytical phase (1). Briefly, the preanalytical phase encompasses all those (prevalently) manually-intensive activities designed for obtaining, handling, transporting, preparing and storing biological samples before testing (2). Reliable evidence, accumulated after decades of research aimed to improve the total quality of the testing process, underpins the notion that the vast majority of problems in laboratory diagnostics are attributable to incorrect or inappropriate preanalytical activities (3).
REFERENCE:
Lippi, Giuseppe, and Camilla Mattiuzzi. “Biological Samples Transportation by Drones: Ready for Prime Time?” Annals of Translational Medicine 4.5 (2016): 92. PMC. Web. 6 Nov. 2017.

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Molecular Viability Testing of UV-Inactivated Bacteria

The polymerase chain reaction (PCR) is effective at detecting bacterial DNA in samples, but it is unable to differentiate viable bacteria from inactivated cells or free DNA fragments. New PCR-based analytical strategies have been developed to address this limitation. Molecular viability testing (MVT) correlates bacterial viability with the ability to rapidly synthesize species-specific ribosomal RNA precursor (pre-rRNA) in response to brief nutritional stimulation. Previous studies demonstrated that MVT can assess bacterial inactivation by chlorine, serum, and low-temperature pasteurization. Here, we demonstrate that MVT can detect inactivation of Escherichia coli, Aeromonas hydrophila, and Enterococcus faecalis cells by ultraviolet (UV) irradiation. Some UV-inactivated E. coli cells transiently retained the ability to synthesize pre-rRNA post-irradiation (generating false-positive MVT results), but this activity ceased within one hour following UV exposure. Viable but transiently undetectable (by culture) E. coli cells were consistently detected by MVT. An alternative viability testing method, viability PCR (vPCR), correlates viability with cell envelope integrity. This method did not distinguish viable from UV-inactivated bacteria under some conditions, indicating that the inactivated cells retained intact cell envelopes. MVT holds promise as a means to rapidly assess microbial inactivation by UV treatment.
IMPORTANCE Ultraviolet (UV) irradiation is increasingly used to disinfect water, food, and other materials for human use. Confirming the effectiveness of UV disinfection remains a challenging task. In particular, microbiological methods that rely on rapid detection of microbial DNA can yield misleading results. This is due to the detection of “remnant” DNA associated with dead microbial cells. This report describes a novel method that rapidly distinguishes living from dead microbial cells after UV disinfection.
REFERENCE:
Kris M. Weigel, et al. Molecular Viability Testing of UV-Inactivated Bacteria. Appl Environ Microbiol. 2017 May 15; 83(10): e00331-17. Prepublished online 2017 Mar 10. Published online 2017 May 1. doi: 10.1128/AEM.00331-17. PMCID: PMC5411506

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#History 1990: Model for inactivation and disposal of infectious HIV and radioactive waste in a BL3 facility

A method is described for autoclaving low levels of solid infectious, radioactive waste. The method permits steam penetration to inactivate biologic waste, while any volatile radioactive compounds generated during the autoclave process are absorbed. Inactivation of radiolabeled infectious waste has been problematic because the usual sterilization techniques result in unacceptable radiation handling practices. If autoclaved under the usual conditions, there exists a high probability of volatilization or release of radioisotopes from the waste. This results in the radioactive contamination of the autoclave and the laboratory area where steam is released from the autoclave. Our results provide a practical method to inactivate and dispose of infectious radioactive waste. For our research, Bacillus pumilus spore strips and vaccinia virus were used as more heat-resistant surrogates of the human immunodeficiency virus (HIV). These surrogates were used because HIV is difficult to grow under most conditions and is less heat tolerant than the surrogates. In addition, B. pumilus has defined cell death values, whereas such values have not been established for HIV. Both B. pumilus and vaccinia virus are less hazardous to work with. The autoclave method is time efficient and can be performed by laboratory personnel with minimal handling of the waste. Furthermore, waste site handlers are able to visually inspect the solid waste containers and ascertain that inactivation procedures have been implemented.

REFERENCE:

Stinson MC, et al. Model for inactivation and disposal of infectious human immunodeficiency virus and radioactive waste in a BL3 facility. Appl Environ Microbiol. 1990 Jan;56(1):264-8.

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Dead or Alive: Molecular Assessment of Microbial Viability

Nucleic acid-based analytical methods, ranging from species-targeted PCRs to metagenomics, have greatly expanded our understanding of microbiological diversity in natural samples. However, these methods provide only limited information on the activities and physiological states of microorganisms in samples. Even the most fundamental physiological state, viability, cannot be assessed cross-sectionally by standard DNA-targeted methods such as PCR. New PCR-based strategies, collectively called molecular viability analyses, have been developed that differentiate nucleic acids associated with viable cells from those associated with inactivated cells. In order to maximize the utility of these methods and to correctly interpret results, it is necessary to consider the physiological diversity of life and death in the microbial world. This article reviews molecular viability analysis in that context and discusses future opportunities for these strategies in genetic, metagenomic, and single-cell microbiology.
REFERENCE:
Cangelosi, Gerard A., and John S. Meschke. “Dead or Alive: Molecular Assessment of Microbial Viability.” Ed. H. L. Drake. Applied and Environmental Microbiology 80.19 (2014): 5884–5891. PMC. Web. 4 Sept. 2017.

Nucleic acid-based analytical methods, ranging from species-targeted PCRs to metagenomics, have greatly expanded our understanding of microbiological diversity in natural samples. However, these methods provide only limited information on the activities and physiological states of microorganisms in samples. Even the most fundamental physiological state, viability, cannot be assessed cross-sectionally by standard DNA-targeted methods such as PCR. New PCR-based strategies, collectively called molecular viability analyses, have been developed that differentiate nucleic acids associated with viable cells from those associated with inactivated cells. In order to maximize the utility of these methods and to correctly interpret results, it is necessary to consider the physiological diversity of life and death in the microbial world. This article reviews molecular viability analysis in that context and discusses future opportunities for these strategies in genetic, metagenomic, and single-cell microbiology.

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