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.

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.

[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.

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.

Management of accidental exposure to Ebola virus in the biosafety level 4 laboratory, Hamburg, Germany.

A needlestick injury occurred during an animal experiment in the biosafety level 4 laboratory in Hamburg, Germany, in March 2009. The syringe contained Zaire ebolavirus (ZEBOV) mixed with Freund's adjuvant. Neither an approved treatment nor a postexposure prophylaxis (PEP) exists for Ebola hemorrhagic fever. Following a risk-benefit assessment, it was recommended the exposed person take an experimental vaccine that had shown PEP efficacy in ZEBOV-infected nonhuman primates (NHPs) [12]. The vaccine, which had not been used previously in humans, was a live-attenuated recombinant vesicular stomatitis virus (recVSV) expressing the glycoprotein of ZEBOV. A single dose of 5 × 10(7) plaque-forming units was injected 48 hours after the accident. The vaccinee developed fever 12 hours later and recVSV viremia was detectable by polymerase chain reaction (PCR) for 2 days. Otherwise, the person remained healthy, and ZEBOV RNA, except for the glycoprotein gene expressed in the vaccine, was never detected in serum and peripheral blood mononuclear cells during the 3-week observation period.

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.

Biohazards symbol: development of a biological hazards warning signal.

The need for a symbol to warn of potential infection hazards became apparent during Public Health Service contract work on the development of containment facilities for virus-leukemia research. A program of direct inquiry and a search of the literature revealed that there was no universally used signal and that scientific and safety organizations concurred in the need for one. Criteria for symbol design were established, and final selection was based on "uniqueness" and "memorability." The National Institutes of Health is recommending use of the symbol as a warning of biological hazard.

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.

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.

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.

Managing potential laboratory exposure to ebola virus by using a patient biocontainment care unit.

In 2004, a scientist from the US Army Medical Research Institute of Infectious Diseases (USAMRIID) was potentially exposed to a mouse-adapted variant of the Zaire species of Ebola virus. The circumstances surrounding the case are presented, in addition to an update on historical admissions to the medical containment suite at USAMRIID. Research facilities contemplating work with pathogens requiring Biosafety Level 4 laboratory precautions should be mindful of the occupational health issues highlighted in this article.

[Biosafety profile of laboratory workers at three education hospitals in Izmir, Turkey]. / Izmir'deki üç egitim hastanesinde laboratuvar çalisanlarinin biyogüvenlik profili.

The laboratory personnel in hospitals are at risk in terms of transmission of various infectious diseases. The aim of this study is to evaluate the knowledge, behavior and attitude of the health personnel who work in one university and two state hospitals in Izmir, Turkey, about biosafety. The study is an observational-sectional study. Participants were selected via random sampling method. The hospitals were visited on workdays determined by the random selection method and all of the personnel (doctor, technician, cleaning-staff) were included to the study. The data were analyzed statistically using Chi square test. Of the 183 participants included in the study, 106 were from Dokuz Eylül University School of Medicine Central Laboratory and 77 were from state hospitals. 62.8% of the participants were female, 37.2% were male and mean age of all was 32.8 +/- 6.9 years. 23.5% of the participants stated that they had previously taken education about biosafety (p= 0.002). It was determined that 91.3% of the participants were wearing gloves and 87.4% of them were wearing lab-coat during laboratory studies. A significant difference was observed between the hospitals in terms of use of gloves (p= 0.004). All the participants stated that they wash their hands and 43% of them indicated that their daily hand wash rate was > or = 10 times. It was determined that 38.3% of the participants consumed food or drinks in the laboratory, however, this rate was statistically significantly less in the university hospital laboratory (p= 0.000). The rate of participants who had been subjected to a microorganism in the last six months was 6.6%. Obedience to the biosafety rules in laboratory will not only provide a safer environment but also improve the quality of work. We believe that the results of this study will serve as a guide for future studies on laboratory biosafety.

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.

[Current biosafety in clinical laboratories in Japan: report of questionnaires' data obtained from clinical laboratory personnel in Japan].

To determine the status of biosafety in clinical laboratories in Japan, we conducted a survey using questionnaires on the biosafety of laboratory personnel in 2004. We obtained data from 431 hospitals (response: 59.5%). Respondents were 301 institutions (70%) having biological safety cabinets (BSCs). BSCs were held in 78% of microbiological laboratories, 7.9% of genetic laboratories, 2.7% of histopathological laboratories, and 1% or less at other laboratories. A clean bench in examination rooms for acid-fast bacilli was applied at 20 hospitals. We found 28 cases of possible laboratory-associated tuberculosis infection, 25 of which were associated with lack of BSC. Other risk factors were immature skills and insufficiently skilled eguipment operation. The frequency of rupture accidents during specimen centrifugation was 67% in dealing with blood and 9.7% in collecting acid-fast bacilli. Half or more accidents were related to inadequate sample tube materials. Technologists were shown to be working on blood collection in many hospitals (75%), and 1,534 events of self-inflicted needle puncture developed in the last 5 years. These results suggest that biosafety systems are woefully lacking or inadequate in clinical laboratories in Japan and must be established at the earliest possible opportunity.

[Biosafety and biosecurity in the medical laboratory. Update and trends]. / Biosiguranta si biosecuritatea în laboratorul medical. Actualitati si tendinte.

Biosafety includes the protective measures against the risks of contamination with pathogen germs in the laboratories that handle pathogens, or stock or manipulate potentially contaminated products, or perform microbiological tests for medical or scientific research purposes, as well as the means of protecting the environment and the human collectivities against hazard contaminations that have as starting point these laboratories. Besides, lately, a new notion emerged, that of biosecurity, which refers to the sum of measures designed to protect workers, environment and population against the loss, theft, use and release in the environment of pathogenic biological agents. The work overviews the present concerns for the regulation of these two notions and the way in which a system for the management of the biological risks in a laboratory that handles pathogens should be documented and implemented. The need for the continuous professional training of the staff and for the establishment of individual and collective responsibilities for preventing biosafety incidents and trespassing biosecurity rules are as well emphasized. The main biosafety measures are pointed out and a series of considerations regarding biosafety and bioterrorism in correlation with the medical laboratory are as well mentioned.

[Biosafety in laboratories concerning exposure to biological agents]. / La biosicurezza nei laboratori per gli esposti ad agenti biologici.

Laboratory workers are exposed to a variety of potential occupational health hazards including those deriving from infectious materials and cultures, radiations, toxic and flammable chemicals, as well as mechanical and electrical hazard. Although all of them are significant, this paper will focus on biological hazards present in clinical and research laboratories. In fact, in spite of numerous publications, guidelines and regulations, laboratory workers are still subject to infections acquired in the course of their researches. This paper describes some aspects that include good microbiological practices (GMPs), appropriate containment equipment, practices and operational procedures to minimize workers' risk of injury or illness.