Biosafety of personnel of microbiological laboratories in the context of the Federal Law of the Russian Federation № 492-FZ of December 30, 2020 «On the biological safety of the Russian Federation¼.

One of the most important requirements for the personnel of microbiological laboratories working with pathogenic and infectious agents is the observance of precautionary measures and the implementation of a set of preventive measures, collectively interpreted as biological safety (biosafety). To a large extent, biosafety problems are also relevant for all clinical laboratories working with biosubstrates, with the potential threat of containing pathogens of bloodborne infections in them. On December 30, 2020, the President of the Russian Federation signed Federal Law № 492 «On the Biological Safety of the Russian Federation¼ (№ 492-FZ), which regulates the basic legal norms and regulation of biosafety issues, as well as a list of measures to prevent the risks of the spread of infections due to accidents, bioterrorist acts and sabotage. The current pandemic of the coronavirus infection COVID-19 has demonstrated, on the one hand, the epidemiological vulnerability of the single world space, and on the other hand, the decisive influence of biological emergencies on the emergence of negative political and economic processes in the world community. In this regard, the issues of ensuring biosafety in the work of microbiological laboratories in the context of protecting personnel and the environment from accidental or unintentional spread of infections are relevant. Working with pathogenic biological agents in microbiological laboratories is constantly associated with the risk of accidents and possible laboratory infection (laboratory-acquired infections) of employees, environmental pollution if the requirements of regulatory documents on biological safety are not met. In accordance with the requirements of № 492-FZ, in order to prevent biological threats, it is necessary to create a system for monitoring biological risks in microbiological laboratories when working with any infected material.

Cellular pathology in the COVID-19 era: a European perspective on maintaining quality and safety.

COVID-19 is a zoonotic viral infection that originated in Wuhan, China, in late 2019. WHO classified the resulting pandemic as a 'global health emergency' due to its virulence and propensity to cause acute respiratory distress syndrome. The COVID-19 pandemic has had a major impact on diagnostic laboratories, particularly those handling cell and tissue specimens. This development carries serious implications for laboratory practice in that safety of personnel has to be balanced against high-quality analysis and timely reporting of results. The aim of this article is to present some recommendations for the handling of such specimens in the preanalytical, analytical and postanalytical phases of laboratory testing and analysis in an era of high COVID-19 prevalence, such as that seen, for example, in the UK, Spain, Italy and France.

Laboratory Biosafety Considerations of SARS-CoV-2 at Biosafety Level 2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen that causes coronavirus disease 2019 (COVID-19), which was first detected in Wuhan, China. Recent studies have updated the epidemiologic and clinical characteristics of COVID-19 continuously. In China, diagnostic tests and laboratory tests of specimens from persons under investigation are usually performed in a biosafety level 2 environment. Laboratory staff may be at greater risk of exposure due to a higher concentration and invasiveness of emerging pathogens. Current infection prevention strategies are based on lessons learned from severe acute respiratory syndrome, expert judgments, and related regulations. This article summarizes biosafety prevention and control measures performed in severe acute respiratory syndrome coronavirus 2 testing activities and provides practical suggestions for laboratory staff to avoid laboratory-acquired infections in dealing with public health emergencies.

Sterilization of Mycobacterium tuberculosis infected samples using methanol preserves anti-tuberculosis drugs for subsequent pharmacological testing studies.

Pharmacokinetic/pharmacodynamic studies of anti-tuberculosis agents in animal models of tuberculosis are hampered by the frequent necessity to perform sample bioanalysis outside the biosafety level-3 environment. Thus, each specimen has to undergo tedious and time-consuming sample sterilization procedures that may also affect drug stability. Here, we tested treatment of Mycobacterium tuberculosis (Mtb) infected samples with methanol to sterilize samples while preserving drug integrity for further pharmacokinetic/pharmacodynamic evaluations. Tissue samples harvested from Mtb infected mice were homogenized, incubated in methanol, and tested for sterility. Once sterility was confirmed, the samples were used to determine concentrations of the anti-tuberculosis drug spectinamide-1599 in lung homogenates using liquid chromatography coupled with mass spectrometry. The results demonstrate that methanol sterilizes tissue samples harvested from Mtb infected mice without altering the integrity of the drug in the tissue.

Proposition of a safe Mycobacterium tuberculosis complex denaturation method that does not compromise the integrity of DNA for whole-genome sequencing.

Whole-genome sequencing plays now a leading role in epidemiologic studies of tuberculosis. DNA extraction of Mycobacterium tuberculosis complex (MTBC) requires complete inactivation of the strains, to be handled for further molecular procedures. In this study we compared two chloroform-based denaturation methods (one with a step of heat killing, one without) to a traditional heat inactivation method. Our results showed that 40% of the strains of MTBC treated by the traditional protocol resulted in a positive culture whereas no culture was observed with the two chloroform-based protocols. The DNA extracts obtained with chloroform-based protocols preparation were successfully used for whole-genome sequencing. We recommend inactivation with our rapid and efficient denaturation method using chloroform without heat killing which met our expectations and biosecurity requirements.

Working Safely with Vaccinia Virus: Laboratory Technique and Review of Published Cases of Accidental Laboratory Infections with Poxviruses.

Vaccinia virus, the prototype Orthopoxvirus, is widely used in the laboratory as a model system to study various aspects of viral biology and virus-host interactions, as a protein expression system, as a vaccine vector, and as an oncolytic agent. The ubiquitous use of vaccinia viruses in laboratories around the world raises certain safety concerns because the virus can be a pathogen in individuals with immunological and dermatological abnormalities, and on occasion can cause serious problems in normal hosts. This chapter reviews standard operating procedures when working with vaccinia virus and reviews published cases of accidental laboratory infections with poxviruses.

Laboratory-acquired Scrub Typhus and Murine Typhus Infections: The Argument for a Risk-based Approach to Biosafety Requirements for Orientia tsutsugamushi and Rickettsia typhi Laboratory Activities.

This study examined the literature on laboratory-acquired infections (LAIs) associated with scrub typhus (Orientia tsutsugamushi) and murine typhus (Rickettsia typhi) research to provide an evidence base for biosafety and biocontainment. Scrub typhus LAIs were documented in 25 individuals, from 1931 to 2000 with 8 (32%) deaths during the preantibiotic era. There were 35 murine typhus LAI reports and no deaths. Results indicated that the highest-risk activities were working with infectious laboratory animals involving significant aerosol exposures, accidental self-inoculation, or bite-related infections. A risk-based biosafety approach for in vitro and in vivo culture of O. tsutsugamushi and R. typhi would require that only high-risk activities (animal work or large culture volumes) be performed in high-containment biosafety level (BSL) 3 laboratories. We argue that relatively low-risk activities including inoculation of cell cultures or the early stages of in vitro growth using low volumes/low concentrations of infectious materials can be performed safely in BSL-2 laboratories within a biological safety cabinet.

Improved Biosafety and Biosecurity Measures and/or Strategies to Tackle Laboratory-Acquired Infections and Related Risks.

Herein, we reviewed laboratory-acquired infections (LAIs) along with their health-related biological risks to provide an evidence base to tackle biosafety/biosecurity and biocontainment issues. Over the past years, a broad spectrum of pathogenic agents, such as bacteria, fungi, viruses, parasites, or genetically modified organisms, have been described and gained a substantial concern due to their profound biological as well as ecological risks. Furthermore, the emergence and/or re-emergence of life-threatening diseases are of supreme concern and come under the biosafety and biosecurity agenda to circumvent LAIs. Though the precise infection risk after an exposure remains uncertain, LAIs inspections revealed that Brucella spp., Mycobacterium tuberculosis, Salmonella spp., Shigella spp., Rickettsia spp., and Neisseria meningitidis are the leading causes. Similarly, the human immunodeficiency virus (HIV) as well as hepatitis B (HBV) and C viruses (HCV), and the dimorphic fungi are accountable for the utmost number of viral and fungal-associated LAIs. In this context, clinical laboratories at large and microbiology, mycology, bacteriology, and virology-oriented laboratories, in particular, necessitate appropriate biosafety and/or biosecurity measures to ensure the safety of laboratory workers and working environment, which are likely to have direct or indirect contact/exposure to hazardous materials or organisms. Laboratory staff education and training are indispensable to gain an adequate awareness to handle biologically hazardous materials as per internationally recognized strategies. In addition, workshops should be organized among laboratory workers to let them know the epidemiology, pathogenicity, and human susceptibility of LAIs. In this way, several health-related threats that result from the biologically hazardous materials can be abridged or minimized and controlled by the correct implementation of nationally and internationally certified protocols that include proper microbiological practices, containment devices/apparatus, satisfactory facilities or resources, protective barriers, and specialized education and training of laboratory staffs. The present work highlights this serious issue of LAIs and associated risks with suitable examples. Potential preventive strategies to tackle an array of causative agents are also discussed. In this respect, the researchers and scientific community may benefit from the lessons learned in the past to anticipate future problems.

Cerebrospinal fluid protein and glucose examinations and tuberculosis:Will laboratory safety regulations force a change of practice?

Cerebrospinal fluid (CSF) protein and glucose examinations are usually performed in chemical pathology departments on autoanalysers. Tuberculosis (TB) is a group 3 biological agent under Directive 2000/54/EC of the European Parliament but in the biochemistry laboratory, no extra precautions are taken in its analysis in possible TB cases. The issue of laboratory practice and safety in the biochemical analyses of CSF specimens, when tuberculosis infection is in question is addressed in the context of ambiguity in the implementation of current national and international health and safety regulations. Additional protective measures for laboratory staff during the analysis of CSF TB samples should force a change in current laboratory practice and become a regulatory issue under ISO 15189. Annual Mantoux skin test or an interferon-γ release assay for TB should be mandatory for relevant staff. This manuscript addresses the issue of biochemistry laboratory practice and safety in the biochemical analyses of CSF specimens when tuberculosis infection is in question in the context of the ambiguity of statutory health and safety regulations.

[Safety in the Microbiology laboratory]. / Seguridad en el laboratorio de Microbiología Clínica.

The normal activity in the laboratory of microbiology poses different risks - mainly biological - that can affect the health of their workers, visitors and the community. Routine health examinations (surveillance and prevention), individual awareness of self-protection, hazard identification and risk assessment of laboratory procedures, the adoption of appropriate containment measures, and the use of conscientious microbiological techniques allow laboratory to be a safe place, as records of laboratory-acquired infections and accidents show. Training and information are the cornerstones for designing a comprehensive safety plan for the laboratory. In this article, the basic concepts and the theoretical background on laboratory safety are reviewed, including the main legal regulations. Moreover, practical guidelines are presented for each laboratory to design its own safety plan according its own particular characteristics.

Using ATP-driven bioluminescence assay to monitor microbial safety in a contemporary human cadaver laboratory.

INTRODUCTION: The objective of this study was to utilize a cost-effective method for assessing the levels of bacterial, yeast, and mold activity during a human dissection laboratory course. Nowadays, compliance with safety regulations is policed by institutions at higher standards than ever before. Fear of acquiring an unknown infection is one of the top concerns of professional healthcare students, and it provokes anti-laboratory anxiety. Human cadavers are not routinely tested for bacteria and viruses prior to embalming. Human anatomy dissecting rooms that house embalmed cadavers are normally cleaned after the dissected cadavers have been removed. There is no evidence that investigators have ever assessed bacterial and fungal activities using adenosine triphosphate (ATP)-driven bioluminescence assays. METHODS: A literature search was conducted on texts, journals, and websites regarding bacterial, yeast, and mold activities in an active cadaver laboratory. Midway into a clinical anatomy course, ATP bioluminescence assays were used to swab various sites within the dissection room, including entrance and exiting door handles, water taps, cadaver tables, counter tops, imaging material, X-ray box switches, and the cadaver surfaces. RESULTS: The results demonstrated very low activities on cadaver tables, washing up areas, and exiting door handles. There was low activity on counter tops and X-ray boxes. There was medium activity on the entrance door handles. CONCLUSION: These findings suggest an inexpensive and accurate method for monitoring safety compliance and microbial activity. Students can feel confident and safe in the environment in which they work.

The hidden face of academic researches on classified highly pathogenic microorganisms.

Highly pathogenic microorganisms and toxins are manipulated in academic laboratories for fundamental research purposes, diagnostics, drugs and vaccines development. Obviously, these infectious pathogens represent a potential risk for human and/or animal health and their accidental or intentional release (biosafety and biosecurity, respectively) is a major concern of governments. In the past decade, several incidents have occurred in laboratories and reported by media causing fear and raising a sense of suspicion against biologists. Some scientists have been ordered by US government to leave their laboratory for long periods of time following the occurrence of an incident involving infectious pathogens; in other cases laboratories have been shut down and universities have been forced to pay fines and incur a long-term ban on funding after gross negligence of biosafety/biosecurity procedures. Measures of criminal sanctions have also been taken to minimize the risk that such incidents can reoccur. As United States and many other countries, France has recently strengthened its legal measures for laboratories' protection. During the past two decades, France has adopted a series of specific restriction measures to better protect scientific discoveries with a potential economic/social impact and prevent their misuse by ill-intentioned people without affecting the progress of science through fundamental research. French legal regulations concerning scientific discoveries have progressively strengthened since 2001, until the publication in November 2011 of a decree concerning the "PPST" (for "Protection du Potentiel Scientifique et Technique de la nation", the protection of sensitive scientific data). Following the same logic of protection of sensitive scientific researches, regulations were also adopted in an order published in April 2012 concerning the biology and health field. The aim was to define the legal framework that precise the conditions for authorizing microorganisms and toxins experimentation in France; these regulations apply for any operation of production, manufacturing, transportation, import, export, possession, supply, transfer, acquisition and use of highly pathogenic microorganisms and toxins, referred to as "MOT" (for "MicroOrganismes et Toxines hautement pathogènes") by the French law. Finally, laboratories conducting researches on such infectious pathogens are henceforth classified restricted area or ZRR (for "Zone à Régime Restrictif"), according an order of July 2012. In terms of economic protection, biosafety and biosecurity, these regulations represent an undeniable progress as compared to the previous condition. However, the competitiveness of research laboratories handling MOTs is likely to suffer the side effects of these severe constraints. For example research teams working on MOTs can be drastically affected both by (i) the indirect costs generated by the security measure to be applied; (ii) the working time devoted to samples recording; (iii) the establishment of traceability and reporting to national security agency ANSM, (iv) the latency period required for staff members being officially authorized to conduct experiments on MOTs; (v) the consequent reduced attractiveness for recruiting new trainees whose work would be significantly hampered by theses administrative constraints; and (vi) the limitations in the exchange of material with external laboratories and collaborators. Importantly, there is a risk that French academic researchers gradually abandon research on MOTs in favor of other projects that are less subject to legal restrictions. This would reduce the acquisition of knowledge in the field of MOTs which, in the long term, could be highly detrimental to the country by increasing its vulnerability to natural epidemics due to pathogenic microorganisms that are classified as MOTs and, by reducing its preparedness against possible bioterrorist attacks that would use such microorganisms.

Análisis bacteriológico de superficies inertes / Bacteriological analysis of inert surfaces

Objetivo: analizar microrganismos presentes en las superficies inertes, que representen un riesgo para la salud de los estudiantes. Métodos: se realizó un estudio observacional, exploratorio y transversal realizado en el periodo febrero- julio de 2012. Se efectuó en un muestreo aleatorio utilizando el método del hisopo y se obtuvieron 72 muestras. Las unidades de análisis fueron mesas, microscopios y charolas por considerarse superficies de mayor contacto con alumnos. Resultados: se encontraron hongos en el 100 por ciento de los cultivos realizados y bacterias en el 66 por ciento. De estas, el 25 por ciento (12) correspondieron a bacterias de flora normal, el 62,5 por ciento (30) a bacterias oportunistas y 12,5 por ciento (6) a bacterias patógenas. Conclusión: las mesas y los microscopios de los laboratorios de enseñanza se encuentran contaminados por hongos y bacterias como Salmonella paratyphi A y Salmonella sp que constituyen un riesgo de infección para los estudiantes que realizan prácticas educativas(AU)

Objective: analyze microorganisms present on inert surfaces which represent a health hazard for students. Methods: an observational cross-sectional exploratory study was conducted from February to July 2012. Random sampling was performed using the swab method. Seventy-two samples were obtained. The study surfaces were tables, microscopes and trays, i.e. the surfaces most commonly touched by students. Results: fungi were found in 100 percent of the cultures. Bacteria were found in 66%. Of the latter, 25 percent (12) were normal flora bacteria, 62.5 percent (30) were opportunistic, and 12.5 (6) were pathogenic. Conclusion: tables and microscopes in teaching laboratories were contaminated with fungi and bacteria such as Salmonella paratyphi A and Salmonella sp., which constitutes an infection hazard for students doing laboratory practice(AU)

[Analysis on status and characteristics of laboratory-acquired vaccinia virus infections cases].

OBJECTIVE: By analyzing the status and characteristics of vaccinia virus laboratory-acquired infections in the bibliographical information, this paper provides relevant recommendations and measures for prevention and control of vaccinia virus laboratory-acquired infections in China. METHODS: Choosing PubMed, Embase, Biosis and SCIE, SSCI, CPCI-S as well as CPCI-SSH covered by Web of Science as the data source, indexing the bibliography of vaccinia virus laboratory-acquired infections, this paper analyzes the information on whether to vaccinate, the occurrence time of symptoms, diseasedparts, symptom characteristics and the disease-causing reasons. RESULTS: The outcome shows that 52. 9% of the cases never get vaccinated, 82.4% engaged in vaccinia virus related researches never get vaccinated in 10 years, 52. 9% get infected by the accidental needlestick in hands during the process of handling animal experiments, 70. 6% of infections occur in the hands and having symptoms after being exposed with an average of 5. 1 days. CONCLUSION: Although it is still controversial that whether or not to be vaccinated before carrying out vaccinia virus related works, it should be important aspects of prevention and control of vaccinia virus laboratory-acquired infections with the strict compliance with the operating requirements of the biosafety, by strengthening personal protection and timely taking emergency measures when unforeseen circumstances occur, as well as providing the research background information to doctors.

Factors influencing biosafety level and LAI among the staff of medical laboratories.

BACKGROUND: The aim of the study was to assess the biological risks of medical laboratory employees with particular focus on laboratory acquired infection (LAI), activities having the greatest risk, accidents with biological material, post exposure procedure, preventive measures and workers' knowledge about biological exposure. MATERIALS AND METHODS: The study involved 9 laboratories. A questionnaire survey was attended by 123 employees and 9 heads of these units with the use of two questionnaires for laboratory workers and the managers. RESULTS: 32.5% of the respondents (40 persons) had an accident at least once. Needlestick or a broken glass injury covered 18.7% respondents (23 persons), while splashing the skin, mucous membranes or conjunctivae related to 22.8% (28 persons). Among the employees who had an accident, only 45% of the respondents (18 persons) reported this to the manager. Microbes dominant in the biological material were known only to 57 respondents (46.3%), less than half could correctly give an example of a disease (57 persons, 46.3%). More than half of the respondents admitted that they do not know all of the possible routes of infection while working in the laboratory (68 persons, 55.3%). CONCLUSIONS: In the study population, a high incidence of accidents was observed, usually during blood sampling and transfer of biological material. Condition of the workers' equipment with personal protective measures and laboratory facilities in devices to reduce the risk of infection and procedures for handling the potentially infectious material should be considered as insufficient. Lack of basic knowledge of the employees about biohazards at workplaces was shown.

[Risk assessment on laboratory biosafety of Leishmania].

OBJECTIVE: To provide the evidence for improving the risk assessment and personal protective equipment and techniques to laboratory staff related to Leishmania. METHODS: The laboratory biosafety of Leishmania was preliminarily assessed based on the biological background information, potential hazards in experimental activities, the risk analyses of laboratory personnel and other relevant factors. RESULTS: The risk assessment on laboratory biosafety of Leishmania was helpful for the establishment of the laboratory standard operating procedure, and was helpful for protecting the staff from infection of Leishmania. CONCLUSION: The risk assessment on laboratory biosafety is important to the safety of laboratory activity related to Leishmania, and is of a great significance to protect the laboratory staff.