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The operator protection factor (OPF) of four biological safety cabinets (BSCs) has been measured under standard and suboptimal conditions.The OPF for one BSC1, two BSC2, and an acid-fast bacilli staining station (AFBSS) was measured using the potassium iodide method for in situ testing of BSCs (CEN12469) over a range of inflow velocities under standard conditions and with common interfering factors (fans, opening doors, and walk pasts).The BSC1 and the AFBSS gave a high level of protection under standard test conditions at all airflows (down to 0.3 and 0.38 m/s, respectively). During interfering processes, the BSC1 and AFBSS gave a high level of protection (OPF10Although BSC2s are capable of giving a high level of performance, this is design dependent and the BSC1 and AFBSS give a more predictable level of performance due to their simpler design. In environments where BSC certification is not possible, they may provide more robust and sustainable primary containment.

OBJECTIVE: The objective of this study was to evaluate whether a Class II type A2 biosafety cabinet in a laboratory could be decontaminated while the laboratory was being fumigated using vaporous hydrogen peroxide or peracetic acid dry fogging. METHODS: To validate decontamination of all parts of the biosafety cabinet, biological indicators were placed at various locations within the biosafety cabinet, including between the pleats of supply and exhaust HEPA filters. To assess whether the operational status of the biosafety cabinet influenced the outcome of its decontamination, fumigation validations were undertaken with the cabinet running and not running. The amount of fumigant and the duration of fumigation remained constant whether the biosafety cabinet was running or not. DISCUSSION: Biosafety cabinet decontamination was successful only when the cabinet was running to facilitate the fumigant’s circulation within the plenums and across the HEPA filters. This study shows both vaporous hydrogen peroxide and peracetic acid dry fogging can be used successfully to decontaminate Class II type A2 biosafety cabinets during laboratory fumigation, provided the biosafety cabinets are operational and running during the fumigation.

Introduction: Keeping a contamination free environment in the laboratory has commonly been achieved by one of two ways: a flame or a biosafety cabinet (BSC). However, it has been frequently observed that these two practices have been combined, where a heat source has been used within the BSC. As flames require flammable gasses and cause hot air to rise, it was hypothesized that these could lead to a loss of BSC containment, as BSCs rely on unidirectional downflow air. Objectives: The objective of this study was to determine whether BSCs can maintain containment when a heat source is operated within the work area. Methods: Several heat sources (Bunsen burner, High Heat Bunsen Burner, Spirit Lamp and Bacti-cinerator) were placed within two sizes of BSCs (4-foot and 6-foot), and smoke was used to visualize airflow disturbances, air cleanliness was measured by particle counting , and aerosol microbiological testing was conducted to ascertain containment. The risk of introducing a flammable gas into a BSC was also calculated. Results: Large flamed Bunsen burners were found to have the most detrimental effects on the ability of the BSC to maintain containment, especially in the center of the work area, while the smaller heat sources were more variable. Containment was completely lost in the 4-foot BSC, whereas the 6-foot BSC was capable of maintaining containment in only a few conditions. The BSC was also calculated to be able to maintain the required volume of flammable gas needed to operate the burners, not taking into consideration unintended leaks. Conclusions: Overall, it was determined that BSCs cannot operate safely and reliably while housing a heat source, as it could cause unexpected contamination of the work or the worker, or BSC ignition or explosion.

Objectives The main aim of this work was to compare the methods of DNA isolation in the moulds of genus Mucorales with special regard to the amount and purity of the DNA acquired. The acquired DNA was then amplified by specific real-time PCR. Design Five DNA extraction procedures were carried out in a Class 2 Biosafety cabinet in a dedicated room with suitable biosafety precautions and appropriate biowaste disposal methods. A total of 6 Mucorales clinical strains were used. Results From the viewpoint of concentration and purity, methods A shown abundant amount of fungal DNA whereas methods E report a pure fungal DNA with R260/280 of 1.7 near the optimal 1.8. The DNA quantity reach statistically difference at ANOVA test with p value 0.0005 Conclusion Overall, the E method was the most efficient method in the extraction of DNA from fungal cultures compared to the other methods considering time, cost, technical expertise, and instrumentation. Use of this assay will allow researchers to obtain DNA from fungi quickly for use in molecular assays

Most laboratory techniques are performed in biosafety cabinets (BSCs) to provide contamination control of the experiments. By using specifically directed airflow and high-efficiency particulate air...

The emergence of highly infectious diseases such as Hepatitis and HIV increasingly drives the importance of health and safety in medical laboratories. This study compared the biosafety and biosecur...

When properly used and adequately maintained, a biological safety cabinet (BSC) provides protection to the laboratory worker, the products, and the environment. Knowledge about proper use of the BS...

Millions of Americans contract food poisoning or are affected by microbial pathogens each year. Rapid, sensitive detection of dilute levels of pathogens in foods, produce, water, and biomanufacturing process samples is key to consumer protection; however, current enrichment methods require as much as a full day to enrich viable bacterial pathogens to detectable levels. Our lab previously demonstrated the ability to concentrate and detect dilute levels of pathogens, within 8 hr, from various food matrices using microfiltration in our continuous cell concentration device (i.e., C3D) with one or two filter modules. This short communication describes the design, materials and construction, layout, and operational characteristics of a four filter module multiplexed system based on a four channel device. Benefits are a 2× greater sample capacity than an equivalent duplex system (achieving the same time to result of less than 8 hr from sample preparation to detection), simpler operation, and a footprint enabling operation inside a biosafety cabinet instead of requiring a BSL-2 room. Flow rate variability through four channels fit within an operational envelope of ±3%; flow rates are reproducible from one run to the next thus ensuring relatively simple, concurrent processing of samples.

Recombinant protein production in the baculovirus expression vector system (BEVS) has emerged as a system of choice for the production of recombinant human proteins for R&D purposes. Scale-up protein production in insect cells past the one or two liter volume generally utilizes disposable cellbag bioreactors that provide a means to scale to the 5-25L range in a single vessel. However, cellbags can be expensive and their use requires capital investment in dedicated rocker platforms and their associated air pumps and exhaust heaters. Additional equipment, such as tube welders and liquid pumps are often also deployed for the sterile transfer of media outside of a biosafety cabinet. Herein it is reported that Sf9, Sf21 and High Five insect cells demonstrate normal growth characteristics when cultured at the 2.5 L level in 3 L Erlenmeyer flasks, or at the 4.5 L level in 5 L Erlenmeyer flasks in standard laboratory shakers. In addition, a direct comparison of the expression levels of four separate proteins at the 4.5 L scale in 5 L flasks versus those at the 5 L scale in 10 L cellbags demonstrates that protein production is equal to, or slightly better, in the flasks versus the cellbags. The adoption of high-volume shake flasks for routine recombinant protein production in insect cells has a number of advantages over disposable bioreactors in terms of ease of use, and equipment and disposables costs.

Biological safety cabinets (BSCs) are the primary means of containment used in laboratories worldwide for the safe handling of infectious microorganisms. They provide protection to the laboratory worker and the surrounding environment from pathogens. To ensure the correct functioning of BSCs, they need to be properly maintained beyond the daily care routines of the laboratory. This involves annual maintenance and certification by a qualified technician in accordance to the NSF/American National Standards Institute 49-2014 Biosafety Cabinetry: Design, Construction, Performance, and Field Certification. Service programs can be direct from the manufacturer or through third-party service companies, but in many instances, technicians are not accredited by international bodies, and these services are expensive. This means that a large number of BSCs may not be operating in a safe manner. In this article, we discuss our approach to addressing the lack of trained and qualified personnel in Thailand who can install, maintain, and certify BSCs in a cost-effective and practical manner. We initiated a program to create both local and regional capacity for repair, maintenance, and certification of BSCs and share our experiences with the reader.

Objective: Chlorine dioxide (CD) gas has been used as a fumigant in the disinfection of biosafety laboratories. In this study, some experiments were conducted to assess the inactivation of spores inoculated on six materials [stainless steel (SS), painted steel (PS), polyvinyl chlorid (PVC), polyurethane (PU), glass (GS), and cotton cloth (CC)] by CD gas. The main aims of the study were to determine the sporicidal efficacy of CD gas and the effect of prehumidification before decontamination on sporicidal efficacy. Methods: Material coupons (1.2 cm diameter of SS, PS, and PU; 1.0 cm×1.0 cm for PVC, GS, and CC) were contaminated with 10 μl of Bacillus subtilis var. niger (ATCC 9372) spore suspension in mixed organic burden and then dried in a biosafety cabinet for 12 h. The spores were recovered by soaking the coupons in 5 ml of extraction liquid for 1 h and then vortexing the liquid for 1 min. Results: The log reductions in spore numbers on inoculated test materials exposed to CD gas [0.080% (volume ratio, v/v) for 3 h] were in the range of from 1.80 to 6.64. Statistically significant differences were found in decontamination efficacies on test material coupons of SS, PS, PU, and CC between with and without a 1-h prehumidification treatment. With the extraction method, there were no statistically significant differences in the recovery ratios between the porous and non-porous materials. Conclusions: The results reported from this study could provide information for developing decontamination technology based on CD gas for targeting surface microbial contamination.

The objective of this study was to evaluate both condensing (wet) and non-condensing (dry) vaporous hydrogen peroxide (VHP) technologies for decontaminating a worst-case scenario Class II biosafety...

Aims: To determine the effectiveness and limitations of ultraviolet (UV) germicidal lamps in biosafety cabinets and pass-boxes, with particular reference to anthrax spores. Methods and Results: An estimated 50 colony forming units (CFUs) of spores of the Sterne strain of Bacillus anthracis were distributed on blood agar plates and on smooth and rough plastic surfaces, glass slides, galvanized metal strips, squares of writing paper, nitrocellulose/cellulose acetate strips, and membrane filters. After drying these were exposed in triplicate to the UV sources of a Class 2 biosafety cabinet and a biosafety level (BSL) 3 pass-box for various time periods. Viable B. anthracis could not be recovered from hard surfaces at 15 minutes (the shortest time period tested) when fully exposed to UV at intensities of i 3.8 W/m2 (the lowest intensity tested) in a Class 2 biosafety cabinet and a BSL3 pass-box. However, the penetrating ability of the UV was found to be extremely limited with even the microstructural elements of nitrocellulose or similar membranes able to substantially shield the spores. The UV failed totally to penetrate thin (66 µm) paper or Petri dish plastic, although it did penetrate plastic sandwich wrap. Conclusions: The UV from germicidal lamps in biosafety cabinets and pass-boxes is rapidly sporicidal for fully exposed anthrax spores on hard surfaces but is readily nullified by the slightest obstacle to direct exposure. Significance and Impact of the Study: The results fill a major information gap for those devising or following protocols for decontamination of items and materials in biosafety cabinets and pass-boxes. Such protocols need to be written around a full comprehension of the data presented here.

During the last three decades, animal cell culturing has been essential for biomedical research and biotechnological activities in general. Along with this increasing importance, biosafety concerns have pointed to the risks of manipulating animal cell cultures for human health and the environment. A maximal reduction of these risks necessitates a thorough risk assessment of the cell cultures used. It involves an evaluation of both the intrinsic properties of the cell culture, including subsequent properties acquired as a result of genetic modification, and the possibility that the cell culture may inadvertently or deliberately become contaminated with pathogens. The latter is a major hazard associated with the manipulation of animal cell cultures, as adventitious agents may be pathogenic and have a better capacity to survive in unfavorable conditions. Consequently, most of the containment measures primarily aim at protecting cells from adventitious contamination. Therefore, a comprehensive evaluation of the risks encountered during the handling of cell cultures should include considerations regarding the type of manipulation as well. As a rule, cell cultures known to harbor an infectious etiologic agent should be manipulated in compliance with containment measures recommended for the etiologic agent. With the exception of very well-characterized cell cultures for which the use of a type II biosafety cabinet depends on the origin of the cells, work with cell cultures from human or primate origin should generally and minimally be performed under containment level 2 using a type II biosafety cabinet. In every case, containment measures should minimize adventitious contamination of the cell cultures and offer a maximal protection of human health and the environment.

Objective: Biosafety is an important element of quality management systems in laboratory practice as it is a measuring tool for compliance with accreditation and certification standards. Our aim was to assess the practices of laboratory personnel towards biosafety measures in their respective laboratories. Methods: The study design was a quantitative and qualitative cross sectional prospective one obtained with the use of a structured questionnaire. Data analysis was done with Epi Info version 3.4.1 and was presented using frequency tables with chi square calculated for different variables with the significant p value set at ≤ 0.05. Results: There were 63 respondents in the study. Private laboratories fared better in an assessment of their biosafety practices. Comparative variables identified as unsafe biosafety practices between public and private laboratories were non availability of Hepatitis B virus antibody testing, p value 0.01, odds ratio 5.4, consuming food in the laboratory, p value 0.00, odds ratio 0.2., non-use of N95 masks p value 0.04, O.R 3.9 and biosafety cabinet use p value 0.05, O.R 2.8. Discussion: Biosafety practices in Nigeria need to be streamlined in accordance with good laboratory practices in order to forestall workplace accidents and promote a safe environment for testing of clinical human samples. Conclusion: The absence of appropriate biosafety policies and practices is one of the challenges facing health care workers in sub Saharan Africa. There is the need for biosafety to be placed in the front burner of issues in laboratory practice in our respective facilities.

The first steps in tissue culture are dating back to the beginning of the nineteenth century when biosafety measures did not yet exist. Later on, animal cell culture became essential for scientific research, diagnosis and biotechnological activities. Along with this development, biosafety concerns have emerged pointing to the risks for human health and in a lesser extent for the environment associated to the handling of animal cell cultures. The management of these risks requires a thorough risk assessment of both the cell cultures and the type of manipulation prior the start of any activity. It involves a case-by-case evaluation of both the intrinsic properties of the cell culture genetically modified or not and the probability that it may inadvertently or intentionally become infected with pathogenic micro-organisms. The latter hazard is predominant when adventitious contaminants are pathogenic or have a better capacity to persist in unfavourable conditions. Consequently, most of the containment measures primarily aim at protecting cells from adventitious contamination. Cell cultures known to harbour an infectious etiologic agent should be manipulated in compliance with containment measures recommended for the etiologic agent itself. The manipulation of cell cultures from human or primate origin necessitates the use of a type II biosafety cabinet. The scope of this chapter is to highlight aspects relevant for the risk assessment and to summarize the main biosafety recommendations and the recent technological advances allowing a mitigation of the risk for the handling of animal cell cultures.

Background: The jet-in-air cell sorters currently available are not very suitable for sorting potentially biohazardous material under optimal conditions because they do not protect operators and samples as recommended in the guidelines for safe biotechnology. To solve this problem we have adapted a cell sorting system to a special biosafety cabinet that satisfies the requirements for class II cabinets. With aid of this unit, sorting can be performed in conformance with the recommendations for biosafety level 2. Methods: After integrating a modified fluorescence-activated cell sorter (FACS) Vantage into a special biosafety cabinet, we investigated the influence of the laminar air flow (LAF) inside the cabinet on side stream stability and the analytical precision of the cell sorter. In addition to the routine electronic counting of microparticles, we carried out tests on the containment of aerosols, using T4 bacteriophage as indicators, to demonstrate the efficiency of the biosafety cabinet for sorting experiments performed under biosafety level 2 conditions. Results: The experiments showed that LAF, which is necessary to build up sterile conditions in a biosafety cabinet, does not influence the conditions for side stream stability or the analytical precision of the FACS Vantage cell sorting system. In addition, tests performed to assess aerosol containment during operation of the special biosafety cabinet demonstrated that the cabinet-integrated FACS Vantage unit (CIF) satisfies the conditions for class II cabinets. In the context of gene transfer experiments, the CIF facility was used to sort hematopoietic progenitor cells under biosafety level 2 conditions. Conclusions: The newly designed biosafety cabinet offers a practical modality for improving biosafety for operators and samples during cell sorting procedures. It can thus also be used for sorting experiments with genetically modified organisms in conformance with current biosafety regulations and guidelines. © 2005 Wiley-Liss, Inc.