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.

A Forgotten Episode of Marburg Virus Disease: Belgrade, Yugoslavia, 1967.

In 1967, several workers involved in poliomyelitis vaccine development and production fell ill at three different locations in Europe with a severe and often lethal novel disease associated with grivets (Chlorocebus aethiops) imported from Uganda. This disease was named Marburg virus disease (MVD) after the West German town of Marburg an der Lahn, where most human infections and deaths had been recorded. Consequently, the Marburg episode received the most scientific and media attention. Cases that occurred in Frankfurt am Main, West Germany, were also described in commonly accessible scientific literature, although they were less frequently cited than those pertaining to the Marburg infections. However, two infections occurring in a third location, in Belgrade, Yugoslavia, have seemingly been all but forgotten. Due in part to their absence in commonly used databases and in part to the fact that they were written in languages other than English, the important articles describing this part of the outbreak are very rarely cited. Here, we summarize this literature and correct published inaccuracies to remind a younger generation of scientists focusing on Marburg virus and its closest filoviral relatives of this important historical context. Importantly, and unfortunately, the three episodes of infection of 1967 still represent the best in-depth clinical look at MVD in general and in the context of "modern" medicine (fully resourced versus less-resourced capacity) in particular. Hence, each individual case of these episodes holds crucial information for health care providers who may be confronted with MVD today.

Survey of Suspected Laboratory-Acquired Infections and Biosafety Practices in Research, Clinical, and Veterinary Laboratories in Karachi, Pakistan.

Laboratory-acquired infections (LAIs), an occupational illness, are defined as all infections acquired through laboratory or laboratory-related activities. A report published in 1898 described an LAI resulting from Corynebacterium diphtheriae being transmitted through mouth pipetting. Despite all efforts, LAIs continue, especially in developing countries like Pakistan, which has been fighting to curb many infectious diseases. As reflected in the published literature, the biosafety culture is severely lacking in many laboratories, and there are no data available from Pakistan on LAIs. Our objective was to ascertain the frequency and rate of LAIs in various labs with versatile portfolios in relation to biosafety and biosecurity practices in Karachi. Ours is a descriptive multicenter cross-sectional study conducted in 30 laboratories located in Karachi from November 2017 to April 2018. Data were collected from laboratories including the university hospital labs, research labs, animal labs, and biomedical labs. Out of 30 facilities, half (n = 15) were clinical/biomedical laboratories, 16.6% (n = 5) were university hospital laboratories, 26.6% (n = 8) were R&D laboratories, and 6.6% (n = 2) were animal laboratories. Needle stick was found to be the most common injury, followed by animal bite/scratch, cut on mucous membrane, falling of personnel, and burn injury.

Laboratory exposure to Coccidioides: lessons learnt in a non-endemic country.

Coccidioides is a primary pathogenic fungus, which infects humans through highly infectious arthroconidia, causing substantial morbidity including life-threatening disseminated infections. Due to the low infectious dose, laboratory personnel might become infected during diagnostic procedures. Accordingly, coccidioidomycosis is reported as the most frequent laboratory-acquired systemic mycosis worldwide. This risk is aggravated in non-endemic countries, where the diagnosis may not be suspected. We report on an inadvertent exposure of 44 persons to Coccidioides posadasii in a clinical microbiology laboratory in Chile, the measures of containment after rapid diagnosis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the lessons learnt in a non-endemic setting.

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.

Outbreak of laboratory-acquired Brucella abortus in Brazil: a case report

Human brucellosis is an occupational disease affecting workers in slaughterhouses, butcher shops and the milk and dairy product industry as well as individuals who work in clinical or research laboratories. We report the first outbreak of a Brucella abortus infection in a Brazilian laboratory and compare the data obtained with reports available in the literature. Exposure was a result of damage to a biological safety cabinet and failure of the unidirectional airflow ventilation system. An epidemiological investigation identified 3 seroconverted individuals, 1 of whom had clinical manifestations and laboratory results compatible with infection at the time of exposure (n=11; attack rate=9.1%).

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.

Outbreak of laboratory-acquired Brucella abortus in Brazil: a case report.

Human brucellosis is an occupational disease affecting workers in slaughterhouses, butcher shops and the milk and dairy product industry as well as individuals who work in clinical or research laboratories. We report the first outbreak of a Brucella abortus infection in a Brazilian laboratory and compare the data obtained with reports available in the literature. Exposure was a result of damage to a biological safety cabinet and failure of the unidirectional airflow ventilation system. An epidemiological investigation identified 3 seroconverted individuals, 1 of whom had clinical manifestations and laboratory results compatible with infection at the time of exposure (n=11; attack rate=9.1%).

Evolution, safety, and highly pathogenic influenza viruses.

Experience with influenza has shown that predictions of virus phenotype or fitness from nucleotide sequence are imperfect and that predicting the timing and course of evolution is extremely difficult. Such uncertainty means that the risk of experiments with mammalian-transmissible, possibly highly virulent influenza viruses remains high even if some aspects of their laboratory biology are reassuring; it also implies limitations on the ability of laboratory observations to guide interpretation of surveillance of strains in the field. Thus, we propose that future experiments with virulent pathogens whose accidental or deliberate release could lead to extensive spread in human populations should be limited by explicit risk-benefit considerations.

Investigation of the first laboratory-acquired human cowpox virus infection in the United States.

BACKGROUND: Cowpox virus is an Orthopoxvirus that can cause infections in humans and a variety of animals. Infections occur in Eurasia; infections in humans and animals have not been reported in the United States. This report describes the occurrence of the first known human case of laboratory-acquired cowpox virus infection in the United States and the ensuing investigation. METHODS: The patient and laboratory personnel were interviewed, and laboratory activities were reviewed. Real-time polymerase chain reaction (PCR) and serologic assays were used to test the patient's specimens. PCR assays were used to test specimens obtained during the investigation. RESULTS: A specimen from the patient's lesion tested positive for cowpox virus DNA. Genome sequencing revealed a recombinant region consistent with a strain of cowpox virus stored in the research laboratory's freezer. Cowpox virus contamination was detected in 6 additional laboratory stocks of viruses. Orthopoxvirus DNA was present in 3 of 20 environmental swabs taken from laboratory surfaces. CONCLUSIONS: The handling of contaminated reagents or contact with contaminated surfaces was likely the mode of transmission. Delays in recognition and diagnosis of this infection in a laboratory researcher underscore the importance of a thorough patient history-including occupational information-and laboratory testing in facilitating a prompt investigation and application of control and remediation measures.

Viral infections in workers in hospital and research laboratory settings: a comparative review of infection modes and respective biosafety aspects.

OBJECTIVES: To compare modes and sources of infection and clinical and biosafety aspects of accidental viral infections in hospital workers and research laboratory staff reported in scientific articles. METHODS: PubMed, Google Scholar, ISI Web of Knowledge, Scirus, and Scielo were searched (to December 2008) for reports of accidental viral infections, written in English, Portuguese, Spanish, or German; the authors' personal file of scientific articles and references from the articles retrieved in the initial search were also used. Systematic review was carried out with inclusion criteria of presence of accidental viral infection's cases information, and exclusion criteria of absence of information about the viral etiology, and at least probable mode of infection. RESULTS: One hundred and forty-one scientific articles were obtained, 66 of which were included in the analysis. For arboviruses, 84% of the laboratory infections had aerosol as the source; for alphaviruses alone, aerosol exposure accounted for 94% of accidental infections. Of laboratory arboviral infections, 15.7% were acquired percutaneously, whereas 41.6% of hospital infections were percutaneous. For airborne viruses, 81% of the infections occurred in laboratories, with hantavirus the leading causative agent. Aerosol inhalation was implicated in 96% of lymphocytic choriomeningitis virus infections, 99% of hantavirus infections, and 50% of coxsackievirus infections, but infective droplet inhalation was the leading mode of infection for severe acute respiratory syndrome coronavirus and the mucocutaneous mode of infection was involved in the case of infection with influenza B. For blood-borne viruses, 92% of infections occurred in hospitals and 93% of these had percutaneous mode of infection, while among laboratory infections 77% were due to infective aerosol inhalation. Among blood-borne virus infections there were six cases of particular note: three cases of acute hepatitis following hepatitis C virus infection with a short period of incubation, one laboratory case of human immunodeficiency virus infection through aerosol inhalation, one case of hepatitis following hepatitis G virus infection, and one case of fulminant hepatitis with hepatitis B virus infection following exposure of the worker's conjunctiva to hepatitis B virus e antigen-negative patient saliva. Of the 12 infections with viruses with preferential mucocutaneous transmission, seven occurred percutaneously, aerosol was implicated as a possible source of infection in two cases, and one atypical infection with Macacine herpesvirus 1 with fatal encephalitis as the outcome occurred through a louse bite. One outbreak of norovirus infection among hospital staff had as its probable mode of infection the ingestion of inocula spread in the environment by fomites. CONCLUSIONS: The currently accepted and practiced risk analysis of accidental viral infections based on the conventional dynamics of infection of the etiological agents is insufficient to cope with accidental viral infections in laboratories and to a lesser extent in hospitals, where unconventional modes of infection are less frequently present but still have relevant clinical and potential epidemiological consequences. Unconventional modes of infection, atypical clinical development, or extremely severe cases are frequently present together with high viral loads and high virulence of the agents manipulated in laboratories. In hospitals by contrast, the only possible association of atypical cases is with the individual resistance of the worker. Current standard precaution practices are insufficient to prevent most of the unconventional infections in hospitals analyzed in this study; it is recommended that special attention be given to flaviviruses in these settings.

[Demodex spp. positivity among laboratory staff, kitchen staff, cleaning workers and nurses working in a state hospital]. / Devlet hastanesi çalisanlarindan laboratuar personeli, mutfak personeli, temizlik isçileri ve hemsirelerdeki Demodex spp. pozitifligi.

OBJECTIVE: This study aimed to determine the prevalence of the types of Demodex spp. among workers in hospitals, where people work collectively in a closed setting for long periods of time. METHODS: In order to determine the parasite prevalence among staff working in the Beydagi State Hospital, including laboratory and kitchen staff, cleaning workers and nurses, defined as risk groups by the infection experts and hospital administration, specimens were collected from their faces using standardized surface skin biopsy (SSSB) and analyzed in parasitology laboratories. Demodex spp. was found in 74.7% of the 95 specimens. The statistical analysis did not reveal any significant differences by jobs, age and gender variables. RESULTS: Moreover, it was observed that positivity was 75.4% for women, and 73.7% for men. Demodex spp. positive staff reported in their histories that they had been diagnosed with rosacea and had pruritus and blushes on their faces from time to time, for which they had not received any medical analyses. Subjects with Demodex spp. were referred to the relevant clinic. CONCLUSION: It was concluded that the high prevalence of the parasite among the hospital staff is associated with inadequate information about Demodex spp. among the staff.

Risks associated with vaccinia virus in the laboratory.

Vaccinia virus (VACV) is used commonly in research laboratories. Non-highly attenuated strains of VACV are potentially pathogenic in humans, and VACV vaccination and biosafety level 2 facilities and protocols are currently recommended for vaccinated laboratory workers in the United States who handle non-highly attenuated strains of the virus. Despite this, laboratory-related VACV exposures continue to occur and a number of recent instances of VACV infection in non-vaccinated laboratory workers have been documented. We provide a discussion of the usage and risks associated with VACV in laboratory research.

Laboratory-acquired vaccinia exposures and infections--United States, 2005-2007.

The last case of naturally acquired smallpox disease, caused by the orthopoxvirus variola virus (VARV), occurred in 1977, and the last laboratory-acquired case occurred in 1978. Smallpox was eradicated largely as the result of a worldwide vaccination campaign that used the related orthopoxvirus, vaccinia virus (VACV), as a live virus vaccine. Routine childhood vaccination for smallpox in the United States was terminated by 1972, but vaccination continues or has been reintroduced for specific groups, including laboratory workers who may be exposed to orthopoxviruses, members of the military, selected health-care workers, and first responders. Severe complications of VACV infection can occur, particularly in persons with underlying risk factors, and secondary transmission of VACV also can occur. VACV is used in numerous institutions for various research purposes, including fundamental studies of orthopoxviruses and use as a vector for the expression of foreign proteins (often antigens or immunomodulators) in eukaryotic cells and animal models. The widespread use of VACV for research has resulted in laboratory-acquired VACV infections, some requiring hospitalization. The current Advisory Committee on Immunization Practices (ACIP) guidelines recommend VACV vaccination for laboratory workers who handle cultures or animals contaminated or infected with nonhighly attenuated VACV strains or other orthopoxviruses that infect humans. This report describes five recent occurrences of laboratory-acquired VACV infections and exposure and underscores the need for proper vaccination, laboratory safety, infection-control practices, and rapid medical evaluation of exposures in the context of orthopoxvirus research.

Pseudo-outbreak of Bordetella bronchiseptica infection associated with contaminated rabbit blood used as a broth culture supplement.

A cluster of clinical isolates of Bordetella bronchiseptica was identified by microbiology laboratory personnel. A clinical and molecular epidemiologic study determined that this cluster represented a pseudo-outbreak due to bacterial contamination of rabbit blood used as a broth culture supplement. This pseudo-outbreak highlights the importance of quality assurance programs in the laboratory.