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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.

Study objective Throughout the coronavirus disease 2019 pandemic, many emergency departments have been using passive protective enclosures ("intubation boxes") during intubation. The effectiveness of these enclosures remains uncertain. We sought to quantify their ability to contain aerosols using industry standard test protocols. Methods We tested a commercially available passive protective enclosure representing the most common design and compared this with a modified enclosure that incorporated a vacuum system for active air filtration during simulated intubations and negative-pressure isolation. We evaluated the enclosures by using the same 3 tests air filtration experts use to certify class I biosafety cabinets: visual smoke pattern analysis using neutrally buoyant smoke, aerosol leak testing using a test aerosol that mimics the size of virus-containing particulates, and air velocity measurements. Results Qualitative evaluation revealed smoke escaping from all passive enclosure openings. Aerosol leak testing demonstrated elevated particle concentrations outside the enclosure during simulated intubations. In contrast, vacuum-filter-equipped enclosures fully contained the visible smoke and test aerosol to standards consistent with class I biosafety cabinet certification. Conclusion Passive enclosures for intubation failed to contain aerosols, but the addition of a vacuum and active air filtration reduced aerosol spread during simulated intubation and patient isolation.

This study aimed to discuss the prevention methods and recommendations from severe acute respiratory syndrome-coronavirus2 (SARS-CoV-2) during the course of pathology laboratory procedures, before macroscopic dissection and during cytological processes. In the light of the latest scientific data, possible virus viability was evaluated on all surfaces and biopsy samples in Hitit University Faculty of Medicine pathology laboratory. In order to continue the pathology and cytology workflow, virus persistence probabilities according to the size of the biopsies, “intraluminal formaldehyde fixation” before luminal organs and especially bowel dissection, consideration prevention methods from contamination in the cytological and histological work process were determined and suggested. Cytological preparations should not be transported in the open after the smear process and should be transported in protective closed containers. Technical handling of cytology specimens should be performed in a Class II biosafety cabinet with appropriate personal protection equipment. In order to accelerate the drying of the smears, it should be avoided to dry by shaking them in the air or drying with the help of air blowing devices. Before the intraoperative consultation, as in the normal process, clinical and pathology correlation should be established and it should be learned whether the patient carries a risk or not. Given the evolving pathology landscape and the high-risk nature of SARS-CoV-2, a clear understanding of the new information necessary for protection methods and implementation in biopsy process has become increasingly important to help guide for pathology practice. The need to organize definite rules within ‘clinic-laboratory procedures and principles’ to prevent unnecessary risks and spreads of the virus and to keep safety in pathology laboratories at the highest level is clear. Healthcare workers must be vaccinated.

The outbreak of Novel Coronavirus Pneumonia SARS-CoV-2 has necessitated the reinforcement of infection control measures in the hospital and laboratory setting. Contact and droplet infection control measures are advised for handling patients diagnosed with COVID-19 and airborne precautions for procedures that generate aerosols.Risk assessment is conducted for all steps of laboratory processes viz. preanalytical, analytical and postanalytical. Standard Precautions must be followed at all times when laboratory staff handle clinical specimens that may contain infectious microorganism. Standard precautions must include hand hygiene along with the use of personal protective equipment (PPE). All samples are collected in appropriate containers and all containers are decontaminated by 62-71% ethanol (alcohol) before transporting them to the laboratory in triple packaging.All samples should undergo initial processing in a biosafety cabinet (BSC). It should be ensured while undertaking all technical procedures that there is minimal formation of aerosols and droplets.All biomedical waste should be disposed as per state and national guidelines. Decontamination includes use of 1% sodium hypochlorite, 62-71% ethanol for surface disinfection or Hydrogen peroxide (0.5%).These laboratory biosafety measures are important to minimise the risk of laboratory transmission of COVID-19 to health care workers.

The gut microbiome and the gut brain axis are potential determinants of Alzheimer’s disease (AD) etiology or severity and gut microbiota might coordinate with the gut-brain axis to regulate behavioral phenotypes in AD mouse models. Using 6-month-old human amyloid precursor protein (hAPP) knock-in (KI) mice, which contain the Swedish and Iberian mutations [APP NL-F (AppNL–F)] or the Arctic mutation as third mutation [APP NL-G-F (AppNL–G–F)], behavioral and cognitive performance is associated with the gut microbiome and APP genotype modulates this association. In this study, we determined the feasibility of behavioral testing of mice in a biosafety cabinet and whether stool from 6-month-old AppNL–G–F mice or AppNL–G–F crossed with human apoE4 targeted replacement mice is sufficient to induce behavioral phenotypes in 4-5 month-old germ-free C57BL/6J mice 4 weeks following inoculation. We also compared the behavioral phenotypes of the recipient mice with that of the donor mice. Finally, we assessed cortical Aβ levels and analyzed the gut microbiome in the recipient mice. These results show that it is feasible to behaviorally test germ-free mice inside a biosafety cabinet. However, the host genotype was critical in modulating the pattern of induced behavioral phenotypes as compared to those seen in the genotype- and sex-match donor mice. Male mice that received stool from AppNL–G–F and AppNL–G–F/E4 donor genotypes tended to have lower body weight as compared to wild type, an effect not observed among donor mice. Additionally, AppNL–G–F/E4 recipient males, but not females, showed impaired object recognition. Insoluble Aβ40 levels were detected in AppNL–G–F and AppNL–G–F/E4 recipient mice. Recipients of AppNL–G–F, but not AppNL–G–F/E4, donor mice carried cortical insoluble Aβ40 levels that positively correlated with activity levels on the first and second day of open field testing. For recipient mice, the interaction between donor genotype and several behavioral scores predicted gut microbiome alpha-diversity. Similarly, two behavioral performance scores predicted microbiome composition in recipient mice, but this association was dependent on the donor genotype. These data suggest that genotypes of the donor and recipient might need to be considered for developing novel therapeutic strategies targeting the gut microbiome in AD and other neurodegenerative disorders.

Processes like single-cell isolation demand an immensely sterile environment and a remarkably magnified plat-form, such as the optical microscope, to analyze the growth of the cell lines and in the precise selection of cells from them. State-of-the-art technologies fail to provide microscopes compatible inside the biosafety cabinets, and therefore the samples are constantly moved from the safety cabinets to the microscope repeatedly for analysis. This, in turn, increases the risk of contamination of the sample as well as the laboratory surroundings. We report the design and development of an automated optical microscope adaptable in the safety cabinet and can be employed for cell isolation processes. Experiments were carried out using the developed imaging system, and results reveal the system’s reliability in cell biology applications.

Covid-19 pandemic is becoming increasingly hazardous to humans, necessitating the use of additional Personal Protective Equipment (PPE) such as N95 Filtering Face-Mask Respirators (FFRs) and face shields to ensure the safety of our frontline personnel. The production of these is becoming an enormous burden, and researchers are attempting to alleviate this stress by designing systems that can make FFRs reuse [1] . Here we present a photocatalytic oxidation sterilization system based on UV-light and synergistically used metal oxide nanoparticles catalyst panels. It is a portable system for fast sterilization of FFRs to suit the growing and urgent requirements. Two indigenous ultraviolet (UV) light sterilization assemblies, having lantern UV-lamps arrangement and peaking at wavelength 254 nm, are fitted within a conventional Class II A2 biosafety cabinet in the developed system to deactivate bacteria and viruses from FFRs, in a portable protective environment. Metal oxide nanoparticle catalysts coated plates are utilized in conjunction with UV lamps in the advanced photocatalytic process. The system is optimized to deliver a UV dose of 1000 mJ/cm 2 for assured decontamination of FFRs in around 5 minutes of treatment time (CDC guidelines for UV dosage for FFR decontamination [2] ). The optimized system is exposed to 10 5 spores (Geobacillus Stearothermophilus) to verify sterilizing efficiency, and we achieved a 5 LOG reduction within 5 minutes of UV-exposure. In addition to bacteria and viruses, it can also remove odors, volatile organic compounds (VOCs), and any other suspended particles from FFRs. One can prepare more than 150 masks for reuse every day using the developed portable system.

Made available in DSpace on 2022-04-29T08:31:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-02 Introduction: The biological safety cabinet (BSC) is a collective protective device used to hold aerosols produced in laboratory procedures. The BSC protects workers, material handling, and the environment. It relies on ultraviolet light (UV) lamps that have germicidal action, altering the nucleic acids of microorganisms. Objective: The objective of the present study was to evaluate the germicidal action of BSC class II type A2 UV light against the culture of two bacterial species with different conditions of exposure to UV light. Methods: For the research, the bacteria Staphylococcus aureus ATCC 25923 and Klebsiella pneumoniae ATCC 10031 were used, at a concentration of 1.5x108 Colony Forming Units/ml (CFU/ml), which were seeded in Petri biplates. Use UV light under different conditions and times. Results: The bacterial species showed the same growth or inhibition profile when subjected to different exposure conditions. Petri dishes with the cover open and protected or not with sterilization packaging showed bacterial inhibition at 15 and 20 minutes. The microorganisms in the biplates protected by the aluminum foil and in the biplates with the cover on, regardless of the time and condition, showed bacterial growth. Conclusion: With the results obtained, it is suggested that the germicidal action of UV light was effective, ensuring adequate decontamination and ensuring quality in laboratory biosafety. Universidade Estadual Paulista Júlio de Mesquita Filho- UNESP Faculdade de Tecnologia de Bauru- FATEC, SP Universidade Estadual Paulista Júlio de Mesquita Filho- UNESP Botucatu. Hospital Estadual de Bauru, SP Universidade Estadual Paulista Júlio de Mesquita Filho- UNESP Universidade Estadual Paulista Júlio de Mesquita Filho- UNESP Botucatu. Hospital Estadual de Bauru, SP

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.

Laboratory training is the cornerstone of science education in higher education. However, in several cases hands-on experimental procedures are not possible, and therefore technology provide alternative educational methods. One of the rapidly evolving technologies, namely Virtual Reality (VR) can offer multiple benefits in laboratory training through the development of simulations and virtual laboratories that support, facilitate, and promote an effective their learning experience. We present an empirical research carried out at the Department of Molecular Biology and Genetics of the Democritus University of Thrace during the winter semester of the academic year 2020-2021. 51 undergraduate students carried out a Virtual Reality activity aiming to train them to the use of a Class II Biosafety Cabinet (BSC) in an immersive virtual environment. Our results show that VR approach was highly and enthusiastically accepted by the students; they reported that they had an authentic learning experience which enabled them to better achieve the learning objectives. However, in some cases symptoms like dizziness and blurry image were reported most likely due to equipment, showing that improvement of the equipment used in VR is needed. Article visualizations

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

Burkholderia pseudomallei is a tier 1 select agent that is associated with laboratory-acquired melioidosis, with international guidelines recommending isolate handling within a class II biosafety cabinet (BSC) in a biosafety level 3 (BSL3) facility. In low-resource settings, this may not be practical; therefore, we aimed to assess the risk of laboratory-acquired melioidosis during routine work. Prior exposure to the organism was determined with a questionnaire and concomitant serology. Of 30 laboratory scientists handling B. pseudomallei on 1,267 occasions outside a biosafety cabinet, no infections were documented and all participants remained seronegative. Additionally, we performed controlled environmental air sampling during 78 laboratory handling events, including plate opening, oxidase testing, and McFarland suspension creation. None of the experiments demonstrated aerosolization of the organism. This study suggests the risk of laboratory-acquired melioidosis is low. However, individual laboratories will need to undertake a risk assessment, including melioidosis endemicity, availability of resources for containment, the nature of routine handling to be undertaken, and the presence of predisposing risk factors for infection in the staff concerned. Additionally, laboratories should take region-specific guidelines into consideration. Further research is required to better inform on the overall risk of infection in the microbiology laboratory.

Cell products consisting of human cells are being actively researched and developed in the areas of regenerative medicine and drug testing platforms. In regenerative medicine, some cell products are used as cell injections, while others are used as a graft with three-dimensional (3D) structures. Using the Kenzan method, it is possible to produce large 3D cell structures that are several centimeters in size. Cyfuse Biomedical K.K. (Cyfuse) is developing 3D cell products as well as equipment for Bio 3D printing. Cyfuse has developed a Bio 3D printer with the following features: (1) it is sufficiently compact to set up in a biosafety cabinet; (2) it allows the flexibility to fabricate several forms of multicellular constructs, in order to expand its potential contribution to research. This chapter introduces the concept of equipment for automatic manufacturing of cell products in regenerative medicine and the functions required for Bio 3D printing using the Kenzan method.

We retrospectively investigated cases of false-positive diagnoses using the BIOFIRE (R) FilmArray (R) meningitis/encephalitis (ME) panel to measure the impact of using a dedicated biosafety cabinet combined with preventive measures to reduce the prevalence of false-positive diagnoses due to pre-analytical in-laboratory contamination. False-positive results were identified by reviewing clinical data, biological parameters and cytology results of cerebrospinal fluid (CSF) samples showing discrepant results between the FilmArray ME panel and routine PCR assays. A total of 327 CSF were analysed over 16 weeks in point-of-care (POC) A and B, over two 8-week periods, periods 1 and 2. The analysis yielded 30 (9 center dot 17%) detection of at least one pathogen including 21/30 (70%) viruses and 9/30 (30%) bacteria. During period 1, POC-A and POC-B manipulated CSF under a non-dedicated hood featuring laminar flow, whereas during period 2, CSFs were manipulated under a dedicated biosafety cabinet without any airflow in POC-A. During period 1, false positives were detected in 3/114 CSF (2 center dot 63%) in POC-A and 1/36 (2 center dot 77%) in POC-B (P = 0 center dot 97); during period 2, false positives were detected in 0/139 CSF (0%) in POC-A and 1/38 (2 center dot 63%) in POC-B (P = 0 center dot 23). All false positives were bacterial. The use of a dedicated cabinet without ventilation along with preventive measures during period 2 in POC-A significantly reduced the number of false-positive results (P = 0 center dot 05). Preventive measures described in this study can mitigate false positives when using PCR-based multiplex assays such as the BIOFIRE FilmArray ME Panel for the diagnosis of meningitis and other infectious diseases.

Despite widespread concern regarding cytokine storms leading to severe morbidity in COVID-19, rapid cytokine assays are not routinely available for monitoring critically ill patients. We report the clinical application of a digital protein microarray platform for rapid multiplex quantification of cytokines from critically ill COVID-19 patients admitted to the intensive care unit (ICU) at the University of Michigan Hospital. The platform comprises two low-cost modules: (i) a semi-automated fluidic dispensing/mixing module that can be operated inside a biosafety cabinet to minimize the exposure of the technician to the virus infection and (ii) a 12–12–15 inch compact fluorescence optical scanner for the potential near-bedside readout. The platform enabled daily cytokine analysis in clinical practice with high sensitivity (

Background: Good laboratory practices in terms of biosecurity and biosafety are essential for the protection of laboratory personnel, the populations and the environment. If not implemented, the risk of pathogen spreading from the laboratory is conceivable. In addition, in current African geopolitical context, the reliable practices such as the protection, control and tracking of valuable biological material will prevent their loss, theft, uncontrolled access and potential misuse. WAHO, with the support of KfW is implementing trainings of trainers on biosecurity in the ECOWAS region in a project called PROALAB. PROALAB team realized that many partners were aiming at implementing in parallel quite similar non internationally certified programs in the ECOWAS region. Therefore, PROALAB decided to organize a meeting of key stakeholders in order to pool efforts, avoid duplication, and harmonize strategies and procedures. Objective: Develop a harmonized strategy and harmonized procedures for the training of health professionals in biosecurity/biosafety and biohazard risk management in ECOWAS member countries to pass internationally certified biosafety and biosecurity examinations. Methods: Programs of each of the institutions were presented and discussed. A regional program on biosecurity/biosafety training was elaborated during group working and a regional program of certification of trainers was discussed and validated. A work plan and timeline were developed through group working. Results: In the ECOWAS region, some countries are more advanced in biosecurity training. Training is generally organized by national biosecurity associations with the support of the IFBA or other partners (WHO, FMx, PHE, etc.). Côte d’Ivoire, Nigeria and Mali seem more advanced and could be solicited to organize training for less-advanced countries. The IFBA certification covers five domains: (1) Biorisk management, (2) biosecurity, (3) biosafety cabinet, (4) biological waste management and (5) biocontainment facility. The technical working group decided consensually that basic training for biosecurity managers in the laboratories should cover the first two domains. More advanced experts advising governments should be trained in the other domains progressively. Progressive training could be built on the 3 levels Nigerian/Ivoirian system of “basic-advanced-consolidated (or expert)” system. Existing module from WHO, FMx, IPD, IPCI and other ECOWAS countries should be adapted to match the international certification needs. Practical training should also be organized in the laboratory by the same institutions that will be allowed to deliver a practical certificate on behalf of WAHO. Conclusion: With the extension of laboratory activities in the context of strengthening the surveillance programs in the ECOWAS region, more expertise on biosecurity is needed. With the support of national biosecurity associations, WAHO could play a role in harmonizing reinforcing training on biosecurity and legislation.

Novel coronavirus infection [coronavirus disease 2019 (COVID-19)] has spread to more than 203 countries of various regions including Africa, America, Europe, South East Asia and Western Pacific. The WHO had declared COVID-19 as the global public health emergency and subsequently as pandemic because of its worldwide spread. It is now one of the top-priority pathogens to be dealt with, because of high transmissibility, severe illness and associated mortality, wide geographical spread, lack of control measures with knowledge gaps in veterinary and human epidemiology, immunity and pathogenesis. The quick detection of cases and isolating them has become critical to contain it. To meet the increasing demand of the diagnostic services, it is necessary to enhance and expand laboratory capabilities since existing laboratories cannot meet the emerging demand. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a BSL-2 (Biosafety Level 2) agent and needs to be handled in biosafety cabinet using standard precautions. This review highlights minimum requirements for the diagnostic laboratories opting testing of material for the diagnosis of COVID-19 and associated biorisk to the individuals and to the community.

With the ongoing novel coronavirus disease 2019 (COVID-19) pandemic, the number of individuals that need to be tested for COVID-19 has been rapidly increasing. A walk-through (WT) screening center using negative pressure booths that is inspired by the biosafety cabinet has been designed and implemented in Korea for easy screening of COVID-19 and for safe and efficient consultation for patients with fever or respiratory symptoms. Here, we present the overall concept, advantages, and limitations of the COVID-19 WT screening center. The WT center increases patient access to the screening clinics and adequately protects healthcare personnel while reducing the consumption of personal protective equipment. It can also increase the number of people tested by 9-10 fold. However, there is a risk of cross-infection at each stage of screening treatment, including the booths, and adverse reactions with disinfection of the booths. These limitations can be overcome using mobile technology and increasing the number of booths to reduce congestion inside the center, reducing booth volume for sufficient and rapid ventilation, and using an effective, harmless, and certified environmental disinfectant. A WT center can be implemented in other institutions and countries and modified depending on local needs to cope with the COVID-19 pandemic.

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...

Objectives Work-related carpal tunnel syndrome (CTS) has been reported in different occupations, including laboratory technicians, so this study was carried out to determine the prevalence and the associated personal and ergonomic factors for CTS among laboratory technicians. Methods A cross-sectional study was conducted among 279 laboratory technicians at King Fahd Hospital, Saudi Arabia, who filled in a self-administered questionnaire, including questions regarding their demographic criteria, occupational history, job tasks, workplace tools, ergonomic factors at work, and symptoms suggestive of CTS. Physical examinations and electrodiagnostic studies were carried out for those who had symptoms suggestive of CTS to confirm the diagnosis. Univariate and multivariate analysis were performed for both personal and physical factors in association with confirmed CTS among laboratory technicians. Results The prevalence of CTS among the laboratory technicians was 9.7% (27/279). The following were the statistically significant risk factors for CTS among them: gender (all cases of CTS were female, P=0.00), arm/hand exertion (OR: 7.96; 95% CI: 1.84-34.33), pipetting (OR: 7.27; 95% CI: 3.15-16.78), repetitive tasks (OR: 4.60; 95% CI: 1.39-15.70), using unadjustable chairs or desks (OR: 3.35; 95% CI: 1.23-9.15), and working with a biosafety cabinet (OR: 2.49; 95% CI: 1.11-5.59). CTS cases had significant longer work duration (17.9 ± 5.6 years) than CTS non-case (11.5 ± 7.4 yeas) with low OR (1.108). Conclusion This study demonstrates some personal and ergonomic factors associated with CTS among the laboratory technicians, including female gender, arm/hand exertion, pipetting, repetitive tasks, working with a biosafety cabinet, and an unadjusted workstation.

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...

Background & Aim Aseptic processing is one of the most important issues in the production of cell-based products which are handled by opening the culture vessels. The practical level of aseptic processing depends on the design of aseptic environment where the equipment for research are installed in the cell manufacturing facility. An aseptic environment where biosafety cabinet (BSC) is installed as the aseptic processing area (Grade A) has vulnerability due to manual operations crossing between aseptic processing area and its surrounding area. Though robustness of an aseptic environment where isolator system is installed is much higher than that in the BSC, the facility cost of the isolator system may be more expensive. On the other hand, the facility with BSC has more lot out risks in production due to biological contamination, resulting in higher manufacturing cost. When the cost of goods for a cell-based product is calculated, it must be considered adding on the required resources with the failure probability of cell manufacturing caused by biological contamination. Methods, Results & Conclusion In this study, biological contamination risks depend on the design of aseptic environment construction has been assessed by introducing the method of work breakdown structure (WBS). Then the failure probability of cell manufacturing caused by biological contamination was simulated with consideration of the possibilities which is based on the particle contamination in the product depending on the equipment usage in facility. It is expected that the established assessment model will be useful for design of cell manufacturing facility.

In order to dilute and effectively remove the bioaerosol in the negative pressure isolation ward, a certain amount of flow rate, i.e., the air change rate, is needed.

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 SPILL RESPONSE method to be used in the event of a biosafety incident.

BIOLOGICAL SPILL RESPONSE method to be used in the event of a biosafety incident.

Objective/background: Guidelines for the manipulation of Mycobacterium tuberculosis (MTB) cultures require a Biosafety Level 3 (BSL-3) infrastructure and accompanying code of conduct. In this study, we aimed to validate and apply detection methods for viable mycobacteria from surfaces in a BSL-3 MTB laboratory. Methods: We evaluated phenotypic (Replicate Organism Detection and Counting [RODAC] plates) and molecular (propidium monoazide [PMA]-based polymerase chain reaction [PCR]) approaches for the detection of viable mycobacteria, as well as the effect of 70% ethanol applied for 5 min for disinfection against mycobacteria. For validation of the method, recovery of serial dilutions of Mycobacterium bovis bacillus Calmette-Guerin from glass slides was measured. Subsequently, we stamped surfaces in and around the biosafety cabinet (BSC) after different technicians had manipulated high bacterial load suspensions for routine drug-susceptibility testing in a Class II BSC. Results: RODAC stamping could detect as few as three bacteria on slides stamped either 5 min or 60 min after inoculation. PMA-based PCR, tested in parallel, did not pass validation. Mycobacteria were still detected after 5-min disinfection with ethanol 70%. In the BSL-3, from 201 RODAC-stamped surfaces, MTB was detected in four: three inside a BSC-on a tube cap and on an operator's gloves-and one outside, on an operator's gown. Conclusion: RODAC plates detect mycobacteria at low numbers of microorganisms. In addition, this method allowed us to show that 70% ethanol does not reliably kill mycobacteria when applied for 5 min to a dried surface, and that MTB bacilli may arrive outside a Class II BSC during routine practice, although the route could not be documented. (C) 2016 Asian-African Society for Mycobacteriology. Production and hosting by Elsevier Ltd. All rights reserved.

Sydney Domville Rowland was a bacteriologist and staff member at the Lister Institute of Preventive Medicine when the First World War broke out in 1914. Following a request to the Director of the Lister Institute to staff and equip a mobile field laboratory as quickly as possible, Rowland was appointed to take charge of No. 1 Mobile Laboratory and took up a temporary commission at the rank of Lieutenant in the Royal Army Medical Corps. On 9 October 1914, Rowland set out for the European mainland and was subsequently attached to General Headquarters in Saint-Omer, France (October 1914-June 1915), No. 10 Casualty Clearing Station in Lijssenthoek, Belgium (June 1915-February 1916, during which period he was promoted Major), and No. 26 General Hospital in Etaples, France (February 1916-March 1917). His research focused on gas gangrene, typhoid fever, trench fever, wound infection and cerebrospinal fever. In February of 1917, while engaged in identifying meningococcal carriers, Rowland contracted cerebrospinal meningitis to which he succumbed at age 44 on 6 March 1917. His untimely death might have been caused by laboratory-acquired meningococcal disease, especially since Rowland9s work with Neisseria meningitidis isolates had extended beyond routine laboratory techniques and included risk procedures like immunisation of rabbits with pathogenic strains isolated from cerebrospinal fluid. Currently, microbiology laboratory workers who are routinely exposed to N. meningitidis isolates are recognised as a population at increased risk for meningococcal disease, for which reason recommended preventive measures include vaccination and handling of isolates within a class II biosafety cabinet.

IntroductionComplete inactivation of virus is crucial before samples are manipulated outside of biological containment areas or general cleaning. There are several control methods that could decrease the risk from viral contamination on surfaces, which include chemical disinfectants, heating sterilisation, and ultraviolet germicidal irradiation depending on the nature and properties of the materials to be sterilised. To date, there are limited studies reporting the effectiveness of physical inactivation methods of dengue virus. Therefore, this study was designed to evaluate the effectiveness of two physical inactivation methods, which are heat- and ultraviolet-inactivation, against dengue virus.Material and methodsAll dengue virus serotypes were subjected to heat treatment at various temperatures and exposed to UV light (wavelength of 250-270 nm) at a distance of approximately 75 cm in a Class II Biosafety cabinet (ESCO) at room temperature. The effectiveness of inactivation methods was tested using viability testing on Vero cells and immunofluorescence assay.ResultsDengue virus can be effectively inactivated by heat treatment at 56°C for at least 30 minutes or at higher temperature. On the other hand, the virus required 45 minutes or longer of ultraviolet light exposure at 75 cm distance from the source to be completely inactivated.ConclusionThe results indicated that DENV can be effectively inactivated using high temperature, i.e. 56°C or above, and UV light irradiation. This result would serve as guidelines in proper decontamination and control of dengue virus in laboratory settings, provided proper conditions are met.

Laboratory testing to support the care of patients with highly infectious diseases may pose a risk for laboratory workers. However, data on the risk of virus transmission during routine laboratory testing conducted using standard personal protective equipment (PPE) are sparse. Our objective was to measure laboratory contamination during routine analysis of patient specimens. Remnant specimens were spiked with the nonpathogenic bacteriophage MS2 at 1.0 × 10 7 PFU/ml, and contamination was assessed using reverse transcriptase PCR (RT-PCR) for MS2. Specimen containers were exteriorly coated with a fluorescent powder to enable the visualization of gross contamination using UV light. Testing was performed by two experienced laboratory technologists using standard laboratory PPE and sample-to-answer instrumentation. Fluorescence was noted on the gloves, bare hands, and laboratory coat cuffs of the laboratory technologist in 36/36 (100%), 13/36 (36%), and 4/36 (11%) tests performed, respectively. Fluorescence was observed in the biosafety cabinet (BSC) in 8/36 (22%) tests, on test cartridges/devices in 14/32 (44%) tests, and on testing accessory items in 29/32 (91%) tests. Fluorescence was not observed on or in laboratory instrumentation or adjacent surfaces. In contrast to fluorescence detection, MS2 detection was infrequent (3/286 instances [1%]) and occurred during test setup for the FilmArray instrument and on FilmArray accessory equipment. The information from this study may provide opportunities for the improvement of clinical laboratory safety practices so as to reduce the risk of pathogen transmission to laboratory workers.

Background & Aim To study the Environment Monitoring (EM) activities in a hospital-based Cell Therapy Manufacturing Facility (mainly processes islet cells and HPC(s), to ensure compliance with FACT (03/2018), ISO14644 (12/2015), validation standards and facility EM SOP. Methods, Results & Conclusion Methods Available EM data from 08/2014 to 12/2017 analyzed, including initial validation EM by vendor. EM are normally performed on a monthly basis, while some months had partial or no EM done due to facility construction, schedule conflicts, or machine error. EM includes particle count (PC), air sampling (AS), contact plate bacterial & fungal (CP). See Image 1 for routine EM locations. CR: Cleanroom; GR: Gown Room; DR: Degown Room; BSC: Biosafety Cabinet. Results In the 41 months, 37 months of particle counts, 40 months of Air Sampling, 38 months of contact plate data were analyzed. Among them, (1) One was In-Process EM, all the others were At Rest EM. The In-Process particle counts were significantly higher than those At-Rest. (2) All critical ISO 5 areas were all within expected ranges for all three EM parameters. (3) Two deviations filed due to high air sampling counts, and both passed after improving operator technique and after facility cleaning. (4) In this 3 year data, particle counts and contact plates results are trending low or stable, within expected range, while Air Sampling results had some fluctuations, see Image 2 and 3. (5) Besides the regular/routine sampling locations, several non-routine sampling locations were also sampled, which detected “mold” in Incubator, “>100 CFU” in freezer, and both repeated “0 CFU” count after equipment decontamination. See Image 3. Conclusion (1) The facility's EM program monitors particulates, microbial contamination, with proper sampling sites, frequency, investigative and corrective actions when specified limits exceeded. (2) All BSC data were satisfactory and safe for cell therapy manufacturing processes. (3) Gown Room and Degown Room were built per ISO 8, and met ISO 7 requirements. (4) More In-Process EM shall be performed to provide baseline data during manufacturing. (5) Non-routine sampling in ISO unclassified areas helps detect suspected/ at risk areas, and help improve the cleanliness of the facility.

Customer demand for convenient food products has led to an increased production of prepackaged and ready-to-eat food products. Most of these products rely mainly on surface disinfection and other traditional approaches to ensure shelf life and safety. Novel processing techniques, such as cold plasma, are currently being investigated to enhance the safety and shelf life of prepacked foods. The purpose of this study was to determine the effects of cold plasma corona discharge on the inactivation of Salmonella on prepackaged, tomato-and-lettuce mixed salads. Two different inoculation methods were evaluated to address cross-contamination of Salmonella from cherry tomatoes to lettuce and vice versa. In separate studies, a sample of either cherry tomatoes (55 g) or romaine lettuce (10 g) was inoculated with a Salmonella cocktail (6.93 ± 0.99 log CFU/mL), placed into a commercial polyethylene terephthalate plastic container, and thoroughly mixed together with its noninoculated counterpart. Mixed salads were allowed to dry in a biosafety cabinet for 1 h. Samples were treated with 35 kV cold plasma corona discharge inside plastic containers for 3 min. Samples were stomached and serially diluted in buffered peptone water and then were plated onto aerobic plate count Petrifilm and incubated for 18 h at 37°C. When lettuce was the inoculated counterpart, log kill of Salmonella was significantly greater on tomatoes (0.75 log CFU/g) compared with lettuce (0.34 log CFU/g) (P = 0.0001). Salmonella was reduced on mixed salad only when lettuce was the inoculated counterpart (0.29 log CFU/g) (P = 0.002). Cold plasma can kill Salmonella in a prepackaged mixed salad, with efficacy dependent on the nature of contamination, direction of transfer, and the surface topography of the contaminated commodity.

Transmission electron microscopy (TEM) is used to observe the ultrastructure of viruses and other microbial pathogens with nanometer resolution. Most biological materials do not contain dense elements capable of scattering electrons to create an image; therefore, a negative stain, which places dense heavy metal salts around the sample, is required. In order to visualize viruses in suspension under the TEM they must be applied to small grids coated with a transparent surface only nanometers thick. Due to their small size and fragility, these grids are difficult to handle and easily moved by air currents. The thin surface is easily damaged, leaving the sample difficult or impossible to image. Infectious viruses must be handled in a biosafety cabinet (BSC) and some require a biocontainment laboratory environment. Staining viruses in biosafety levels (BSL)-3 and -4 is especially challenging because these environments are more turbulent and technicians are required to wear personal protective equipment (PPE), which decreases dexterity. In this study, we evaluated a new device to assist in negative staining viruses in biocontainment. The device is a capsule that works as a specialized pipette tip. Once grids are loaded into the capsule, the user simply aspirates reagents into the capsule to deliver the virus and stains to the encapsulated grid, thus eliminating user handling of grids. Although this technique was designed specifically for use in BSL-3 or -4 biocontainment, it can ease sample preparation in any lab environment by enabling easy negative staining of virus. This same method can also be applied to prepare negative stained TEM specimens of nanoparticles, macromolecules and similar specimens.

Background The Spanish National Institute of Security and Hygiene at Work has just published a new technical note ‘Hazardous drugs: prevention measures for their preparation and administration’, based on the list of biohazardous drugs published by the National Institute for Occupational Safety and Health (NIOSH). Purpose To determine the degree of implementation of the document recommendations in the pharmacy unit of our hospital. In particular, to quantify the number of drugs whose preparation meets the standards set out in the technical note with the intention to improve the adequacy of the pharmacy unit protocols. Material and methods The drugs available in the pharmacy unit that are included in the technical note ‘Biohazardous drugs’ are classified into the three categories: biosafety cabinet (BSC) I and II and reproductive risk (RR). Results Presently, 121 of 213 drugs included in the technical note are available in our hospital pharmacy. Of these, 53 (43.8%) should be handled into BSC II according to the standards. The preparation of 48 (90.6%) hazardous drugs was performed in accordance with the recommended prevention measures. Only 5 drugs (9.4%) (phenytoin, leuprorelin, risperidone, tacrolimus, triptorelin) were not handled according to the standards. 61/121 drugs should be handled in BSC I (43 only BSC I and 18 BSC I and RR). None of these drugs were prepared in accordance with the recommended prevention measures. From the 7 drugs with a proven RR, 5 (71.4%) were not handled according to the standards. Conclusion At present, 90.6% of biohazardous drugs BSC II, none of biohazardous drugs BSC I and 28.6% with RR were handled according to the standards set in the technical note, which leads us to conclude that implementation of the recommendations reported is still low in our hospital. Setting up multidisciplinary groups of professionals, including labour union representatives and experts in occupational health and safety, would undoubtedly be helpful to improve the implementation of these recommendations in the healthcare system. The resulting increase in the workload of the pharmacy units is a fact that must be taken into account by the authorities in each centre. References and/or acknowledgements Technical note. No conflict of interest

Sample preparations for transmission electron microscopy of virulent Mycobacterium tuberculosis are usually performed with chemical fixation using glutaraldehyde (GA) in a biosafety area followed by post-fixation with aqueous osmium tetroxide (OT) in a conventional laboratory outside the biosafety area. Freeze-substitution with osmium-acetone (OA) at ultralow temperature (− 85 °C) has been shown to provide high quality final images and preserves cellular structures intact. However, some preparation procedures for freeze-substitution often require large fixed devices for freezing in a special laboratory. We have reported a novel freeze-substitution preparation method that can be performed using a portable device in a biosafety cabinet at biosafety level (BSL) 3 areas. Here, as a next step, we examined whether images obtained from rapid freeze-substitution (RFS) after fixation with glutaraldehyde (GA > RFS) are of comparable quality to those obtained using standard RFS. GA > RFS provided excellent preservation of mycobacterial cell ultrastructure, including visualization of cytoplasmic ribosomes, DNA fibers, and the outer membrane. The average number of ribosomes per cubic micrometer counted on RFS and GA > RFS was not significantly different (6987.8 ± 2181.0 and 6888.9 ± 1799.3, respectively). These values were higher, but not significantly so, than those obtained using conventional chemical fixation (5018.7 ± 2511.3). This procedure may be useful for RFS preparation of unculturable mycobacteria strains or virulent strains isolated in laboratories that cannot perform RFS.

In order to dilute and effectively remove the bioaerosol in the negative pressure isolation ward, a certain amount of flow rate, i.e., the air change rate, is needed.

Soil columns have been utilized in hydrology to study vertical solute transfer through porous material for decades. Soil columns are typically designed as open tubes with soil held in place with meshing. While this open design is sufficient for non-hazardous particles, it is not ideal for hazardous biological contaminants that may be harmful to humans. The design of this study features a closed soil column system for use with potentially hazardous biological components. The apparatus is comprised of a mist nozzle, flow-reducing cap, and meshing to simulate rainfall on each soil column. After percolating through the soil, water and contaminants pass through a funnel coupling and discharge tube into a collection container. For additional safety, the soil column design fits within a standard biosafety cabinet for use with hazardous contaminants. Its modular design allows for simple maintenance, water flowrate adjustment, and versatility that encourages use in multiple applications. These soil columns were created to study the vertical flow of pathogens, pesticides, and other biological agents. Further experimentation with various hazardous components will develop a better understanding of their fate and transport in soil. This paper details the construction processes and testing methods to validate the system&rsquo, s ability to replicate a desired flowrate, which is a precursor to studying the vertical transport of pathogens and other agents through soil.

Background The aim of this study was to assess public and private medical diagnostic laboratories in Nigeria for the presence of biosafety equipment, devices, and measures. Methods A total of 80 diagnostic laboratories in biosafety level 3 were assessed for the presence of biosafety equipment, devices, and compliance rate with biosafety practices. A detailed questionnaire and checklist was used to obtain the relevant information from enlisted laboratories. Results The results showed the presence of an isolated unit for microbiological work, leak-proof working benches, self-closing doors, emergency exits, fire extinguisher(s), autoclaves, and hand washing sinks in 21.3%, 71.3%, 15.0%, 1.3%, 11.3%, 82.5%, and 67.5%, respectively, of all laboratories surveyed. It was observed that public diagnostic laboratories were significantly more likely to have an isolated unit for microbiological work ( p = 0.001), hand washing sink ( p = 0.003), and an autoclave ( p ≤ 0.001) than private ones. Routine use of hand gloves, biosafety cabinet, and a first aid box was observed in 35.0%, 20.0%, and 2.5%, respectively, of all laboratories examined. Written standard operating procedures, biosafety manuals, and biohazard signs on door entrances were observed in 6.3%, 1.3%, and 3.8%, respectively, of all audited laboratories. No biosafety officer(s) or records of previous spills, or injuries and accidents, were observed in all diagnostic laboratories studied. Conclusion In all laboratories (public and private) surveyed, marked deficiencies were observed in the area of administrative control responsible for implementing biosafety. Increased emphasis on provision of biosafety devices and compliance with standard codes of practices issued by relevant authorities is strongly advocated.

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.

The early use of continuous cell lines (CCLs) for the manufacture of biological products is represented by the manufacture of foot-and-mouth disease vaccine in the Syrian hamster cell line baby hamster kidney (BHK), the production of interferon from the B-lymphoblastoid cell line Namalwa, and the introduction of monoclonal antibodies from hybridoma cells. Long-standing experience has shown that contamination with pathogenic agents is the most important hazard and merits careful assessment of safety precautions. The culture types, in order of decreasing risk, are primary cell cultures; CCLs; and intensively characterized cells, including human diploid fibro-blasts. Most national guidelines recommend that human and other primate cells be handled using biosafety level 2 practices and containment and that all work be performed in a biosafety cabinet (BSC). For airfreight, cell cultures have been classified as ‘’diagnostic specimens’’ under the International Air Transport Association (IATA) regulations. In many cases gene therapy concepts are based on viral vectors (e.g., adenoviruses and retroviruses). This chapter describes safety measures for large-scale production of biologicals, and focuses on characterization of cell lines. PCR is now commonly used to identify cell lines and to detect both cross contamination by other cell lines and contamination by adventitious agents or in combination with other methods. Safety testing of cells used in the manufacture of biologicals is usually performed with procedures accredited to good laboratory practice.

This chapter provides mechanisms for the evaluation of the components of a biosafety program and gives examples of tools that can be used for this process. A biosafety program may include blood-borne pathogens, infectious materials, diagnostic specimens, recombinant-DNA (rDNA) oversight, management of biosafety cabinet (BSC) certification, and/or the select agent program. The most common method used to evaluate a biosafety program is an audit of the laboratory. Biosafety manager, self-inspection, safety generalist, outside consultant and regulatory agency are explained in this chapter. One logical way to evaluate a component of the biosafety program is through review of records and documentation rather than a site visit. In many institutions the biosafety program manages the BSC certification program. The chapter discusses annual reverification of biosafety level 3 (BSL-3) laboratories. Inspection should be scheduled well in advance to ensure that laboratory experiments are not in progress, since the annual reverification focuses on facility operations and is not a review of laboratory procedures. A BSL-3 facility should be reverified on an annual basis to ensure that it continues to meet the BSL-3 criteria stated in Biosafety in Microbiological and Biomedical Laboratories (BMBL).

Any discussion of biological safety within undergraduate basic science and clinical teaching laboratories should be prefaced by several important caveats. First, the foundation of any science laboratory experience should include a solid understanding of safety. Practicing safety teaches responsibility and respect for life and property. Regardless of unique institutional conditions and oversight agencies, adherence to common safety and biosafety practices demonstrates a good-faith effort to students, parents, faculty, administrators, and even accreditors, that academic laboratory users are valued. Additionally, most safety practices were developed in response to documented need and thus serve to mitigate hazards. At its core, the teaching laboratory should be an environment where students are challenged by the science of microbiology, not by fear of infection. Furthermore, as the science of microbiology evolves, time-tested biosafety practices continue as a microbiology legacy, passed down to subsequent generations of microbiologists as resume staples.

SummaryAims: Vaccination by aerosol inhalation can be used to efficiently deliver antigen against HPV to mucosal tissue, which is particularly useful in developing countries (simplicity of administration, costs, no need for cold chain). For optimal immunological response, vaccine particles should preferentially be delivered to proximal bronchial airways. We aimed at quantifying the deposition of inhaled particles in central airways and peripheral lung, and to assess administration biosafety. Participants, methods: 20 healthy volunteers (13W/7M, aged 24±4y) performed a 10-min free-breathing inhalation of 99mTc-stannous chloride colloid aerosol (450 MBq) in a buffer solution without vaccinal particles using an ultrasonic nebulizer (mass median aerodynamic diameter 4.2 μm) and a double mask inside a biosafety cabinet dedicated to assess environmental particle release. SPECT/CT and whole-body planar scintigraphy were acquired to determine whole-body and regional C/P distribution ratio (central-to-peripheral pulmonary deposition counts). Using a phantom, SPECT sensitivity was calibrated to obtain absolute pulmonary activity deposited by inhalation. Results: All participants successfully performed the inhalation that was well tolerated (no change in pulmonary peak expiratory flow rate, p = 0.9). It was environmentally safe (no activity released in the biosafety filter.) 1.3±0.6% (range 0.4–2.6%) of the total nebulizer activity was deposited in the lungs with a C/P distribution ratio of 0.40±0.20 (range 0.15–1.14). Conclusion: Quantification and regional distribution of inhaled particles in an aerosolized vaccine model is possible using radioactive particles. This will allow optimizing deposition parameters and determining the particles charge for active-particles vaccination.