Biological Safety Cabinet Research Articles

A comparison of air sampling methods for Clostridium difficile endospore aerosol

The airborne dissemination of Clostridium difficile (C. difficile) endospores (spores) in healthcare environments is documented in multiple studies. Once airborne, spores have the potential for transport on air currents to other areas. This study compared the methods in the collection of C. difficile spore aerosol. This study determined the relative efficiency of commonly used bioaerosol air sampling methods when characterizing airborne C. difficile spore concentrations. Air samplers evaluated in this study were the AirTrace slit-to-agar impactor, AGI-30 impinger, SKC BioSampler impinger, and a 47-mm mixed cellulose ester (MCE) filter cassette. Non-toxigenic C. difficile spores were nebulized into an enclosure contained in a biological safety cabinet. Side-by-side air samples were drawn from the enclosure. The slit-to-agar impactor, successfully used in previous studies to collect airborne spores, served as the reference method. Relative efficiency for the 47-mm MCE filter cartridge was higher than the slit-to-agar impactor (mean 136.6%, 95% CI 124.7–148.5%). Efficiencies of the impingers were similar and were low (mean 4.13%, 95% CI 2.27–5.99%). Impingers failed to maintain culturability of C. difficile spores during sampling. This study is the first to compare the efficiencies of commonly used bioaerosol sampling methods to collect airborne C. difficile spores. Filter air sampling provided the greatest collection of airborne spores. Slit-to-agar air sampling may underestimate the number of airborne spores present. Impinger air sampling could significantly underestimate the actual number of airborne C. difficile spores present or fail to detect airborne spores.

Validation of cleaning procedures used in an Italian Hospital Pharmacy for antineoplastic drug decontamination: a new tool for industrial hygiene.

Background:Current Italian regulations and procedures for surface decontamination of antineoplastic drugs (ADs) are not clear. Therefore, most hospital pharmacies follow internal procedures as an interpretation of the recommended handling guidelines.Objectives:Our study compared 7 different cleaning procedures after controlled contamination of the work surface of a biological safety cabinet workbench in an Italian hospital oncology pharmacy (HOP) to determine which of them is more efficient and practical. Moreover, in order to approximate operative routine and improve risk awareness, cleaning procedures were carried out by the personnel that usually operate in the HOP.Methods:Measured quantities, i.e. a drop (100 μL) of 5-FluoroUracil, IPhosfamide, CycloPhosphamide and Gemcitabine, were deposited on the work surface within precisely delimited areas. Following the wipe-test analysis using UPLC-MS/MS, the cleaning efficacy was calculated based on the ratio of the residual concentration of the AD, after the cleaning procedure, to the concentration of each AD before the procedure.Results:Tested cleaning procedures were: 1) Hypo-Chlor®, hot water and Farmecol70®; 2) Hypo-Chlor® and hot water; 3) Farmecol70®; 4) Surfa’Safe SH® and hot water; 5) Amuchina® 10%, hot water and Farmecol70®; 6) Incidin® Oxyfoam and hot water; 7) liquid Marseille soap, hot water and Farmecol70®. Within the studied HOP, the Marseille soap was evaluated to be the optimal choice due to its efficacy, low cost, and the very short contact time needed before rinsing.Discussion:The application of the protocol for procedure validation suggested here could be used in every HOP as a reliable industrial hygiene tool to demonstrate the validity of the chosen cleaning procedure.

Draft Genome Sequences of Three Novel Staphylococcus arlettae Strains Isolated from a Disused Biological Safety Cabinet.

The genome sequences of three new strains of Staphylococcus arlettae named Bari1, Bari2, and Bari3 are presented. The strains exhibited tolerance to hexavalent chromium ions. An sprC gene encoding a putative chromium transporter was present in each of the three draft genome sequences.

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

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

Closed-system drug-transfer devices plus safe handling of hazardous drugs versus safe handling alone for reducing exposure to infusional hazardous drugs in healthcare staff.

Closed-system drug-transfer devices plus safe handling of hazardous drugs versus safe handling alone for reducing exposure to infusional hazardous drugs in healthcare staff.

バイオクリーン施設への安全キャビネットの適用に関する研究 （その2）空調気流や扉開閉で生じる気流が安全キャビネット内部の清浄度に及ぼす影響

バイオクリーン施設への安全キャビネットの適用に関する研究 （その2）空調気流や扉開閉で生じる気流が安全キャビネット内部の清浄度に及ぼす影響

Francisella tularensis: Understanding Reported Occupational Exposures and Laboratory Methods Used for the Identification of Francisella tularensis.

Francisella tularensis, the causative agent of tularemia, is a HHS Tier 1 select agent. Tularemia is the most commonly reported human and animal infection caused by a bacterial select agent in the United States. Because of the rarity of disease, low clinical suspicion, and the organism's low infectious dose, F. tularensis poses a hazard for unsuspecting laboratorians, particularly those who handle cultures outside a biological safety cabinet or without use of appropriate personal protective equipment (PPE). We examined Form 4s and Form 3s submitted to the Federal Select Agent Program between 2011 and 2015 to assess laboratory methods used in the identification of F. tularensis and categorize reported occupational exposures. Culture, which is used in a confirmatory identification, was the primary method used in clinical laboratories. Reported occupational exposures in clinical, veterinary, and reference laboratories occurred at a rate of 33.8, 14.0, and 0.4/100 isolates, respectively. The number of exposed workers in clinical, reference, veterinary, and research laboratories was 3.2, 2.4, 5.1, and 0.9 workers per reported incident, respectively. Most reported occupational exposures occurred in clinical laboratorians working on the bench at BSL-2 conditions with isolated cultures with no suspicion that the organism was F. tularensis; the fewest occurred in research laboratories at BSL-3 where occupational exposures were prevented by prior knowledge that the organism was F. tularensis and the PPE that was used in these laboratories.

Comparison of Decontamination Efficacy of Cleaning Solutions on a Biological Safety Cabinet Workbench Contaminated by Cyclophosphamide

Several studies have compared cleaning procedures for decontaminating surfaces exposed to antineoplastic drugs. All of the cleaning products tested were successful in reducing most of the antineoplastic drug quantities spilled on surfaces, but none of them completely removed residual traces. To assess the efficacy of various cleaning solutions for decontaminating a biological safety cabinet workbench exposed to a defined amount of cyclophosphamide. In this pilot study, specific areas of 2 biological safety cabinets (class II, type B2) were deliberately contaminated with a defined quantity of cyclophosphamide (10 μg or 107 pg). Three cleaning solutions were tested: quaternary ammonium, sodium hypochlorite 0.02%, and sodium hypochlorite 2%. After cleaning, the cyclophosphamide remaining on the areas was quantified by wipe sampling. Each cleaning solution was tested 3 times, with cleaning and wipe sampling being performed 5 times for each test. A total of 57 wipe samples were collected and analyzed. The average recovery efficiency was 121.690% (standard deviation 5.058%). The decontamination efficacy increased with the number of successive cleaning sessions: from 98.710% after session 1 to 99.997% after session 5 for quaternary ammonium; from 97.027% to 99.997% for sodium hypochlorite 0.02%; and from 98.008% to 100% for sodium hypochlorite 2%. Five additional cleaning sessions performed after the main study (with detergent and sodium hypochlorite 2%) were effective to complete the decontamination, leaving no detectable traces of the drug. All of the cleaning solutions reduced contamination of biological safety cabinet workbenches exposed to a defined amount of cyclophosphamide. Quaternary ammonium and sodium hypochlorite (0.02% and 2%) had mean efficacy greater than 97% for removal of the initial quantity of the drug (107 pg) after the first cleaning session. When sodium hypochlorite 2% was used, fewer cleaning sessions were required to complete decontamination. Further studies should be conducted to identify optimal cleaning strategies to fully eliminate traces of hazardous drugs.

The incidence of musculoskeletal system occupational diseases among tuberculosis laboratory workers

Objectives. Health care workers are exposed to various kinds of professional risks like needle stick injuries, lower back and back problems, allergies, violence and stress. Health care workers in tuberculosis laboratory are exposed to both infection and musculoskeletal system occupational disease risks because of using Class II, Type B biological safety cabinets and laboratory vortex equipment. This study was carried out to determine incidence rates and causes of the musculoskeletal system occupational diseases among health care workers in tuberculosis laboratories of two State hospitals in Ankara. Methods. Study population was composed of 16 laboratory workers in tuberculosis laboratories of two State hospitals in Ankara. Data was collected using a questionnaire. Results. Thirteen (81.2%) laboratory workers were male and three (18.8%) were female. The mean age of the subjects was 40.1 ± 7.0 years and average duration of occupation of subjects was 208.3 ± 11.6 month. According to the data, of the 16 personnel, 10 (62.5%) had occupational disease. Incidence rates of occupational diseases were as follows; 62.5% shoulder pain, 25.0% wrist pain and 18.8% elbow pain. Duration of their occupation was significantly associated with elbow pain ( p = 0.037). There was a significant relationship between hand and wrist pain and smoking ( p = 0.042). Seventy-five per cent of laboratory workers did not think they had enough information on occupational diseases, and 68.8% of them wanted to have information about occupational diseases. Discussion. The most prevalent occupational diseases among the tuberculosis laboratory workers in our sample were shoulder, elbow and wrist pains. Duration of occupation and smoking were associated with the incidence of occupational diseases.

Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory.

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented—one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

Sensitivity tests of biological safety cabinets' contaminant contention to variations on indoor flow parameters in biosafety level laboratories

Laboratory buildings demand from 4 to 6 times more energy use than conventional applications, and near 50% of this energy use is due to its ventilation system. Previous studies have demonstrated that the strategy of increasing ventilation rates, in mixing ventilation mode, has limited effectiveness on airborne infection control and that the local airflow patterns are more important than a global ventilation rate for that purpose. The primary objective of this research was to assess the sensitivity of contaminant contention to variations on indoor flow parameters, for biosafety level (BSL-Biosafety level) laboratories. Selected testing scenarios were modeled, and the only contaminant source was located inside the biological safety cabinet (BSC-Biological safety cabinets). Numerical modeling, using computational fluid dynamics was performed, in order to assess the sensitivity of BSC's contaminant contention to different values on: generation rate; room thermal load; downflow turbulence; inflow velocity; and room air change rate. Another objective of this research was to assess the time scale related to contaminant transport from the BSC interior to the operator respiration zone, after a concentration build up has taken place inside the BSC. The results showed that BSC's contaminant contention is dependent on room indoor flow pattern, room turbulence level and inflow velocity. The use of higher values of ventilation rates in biosafety level laboratories should be analyzed with caution, because increasing the ventilation rate, with mixed ventilation, decreases the BSC's contaminant contention, due to the enhanced turbulent level, and may increase personnel exposure to hazardous contaminants that leak from the BSC.

An evaluation of chemotherapy drug preparation process in hospitals in Turkey - A pilot study.

To identify the practice differences among chemotherapy units and the opinions of pharmacists on safety precautions for handling antineoplastic drugs in hospitals. The survey was administered to hospital pharmacists in Turkey. The pharmacists who work in chemotherapeutic drug preparation units were asked to participate if they are volunteered. The participating pharmacists were asked questions about the type of chemotherapy units, order and verification process of chemotherapy protocols and potential shortcomings of chemotherapy preparation. The questions were initially assessed in a small sample of pharmacists and subsequently revised and refined. The filled and returned questionnaires were accepted as given consent. A total of 97 questionnaires were delivered and 40 (41%) were returned by pharmacists. The median age and practicing years in hospitals of the pharmacists were 38.0 and 6.25 years, respectively. Fully automated chemotherapy units were used by 42.5% of respondents, whereas 30% prepare chemotherapeutics manually; furthermore, 87.5% provide chemotherapeutic drug preparations (admixtures) less than 120 per day. With regard to the drug preparation process, more than half of the respondents 'always' use protective equipment without foot covers (32.3%). Almost 90% of respondents 'always' use a coat, mask and biological safety cabinets; however, 20% of respondents 'often' use closed-system drug transfer sets. More than 50% of pharmacists expressed that they have sufficient knowledge on cytotoxics and the risk of exposure, protective precautions, selection and usage of personal protective equipment and correct storage of cytotoxics. Pharmacists acknowledge sufficient understanding of safe preparation and handling of chemotherapeutics, but reported practices by pharmacists in hospitals are not consistent with published recommendations. Providing theoretical and practical educations for pharmacists and developing standard procedures for preparing chemotherapy may be beneficial in improving the preparation and use of antineoplastics in Turkey.

Performance evaluation of the compounding robot, APOTECAchemo, for injectable anticancer drugs in a Japanese hospital

BackgroundThe accuracy, safety and feasibility of, the compounding robot APOTECAchemo were evaluated in the clinical practice of Japan.MethodsAccuracy and precision of robotic preparations by APOTECAchemo was evaluated in 20 preparations of fluorouracil (FU) and cyclophosphamide (CPA) infusions by four pharmacists. Environmental and product contaminations with FU and CPA were evaluated by wipe testing. Robotic performance was compared with manual preparation in a biological safety cabinet. The number of robotic products, total compounding time and total pre-reconstitution time of lyophilized drugs between January 1, 2014 to December 31, 2015 were investigated.ResultsRobotic preparation resulted more accurate and precise (mean absolute dose error and coefficient of variation were 0.83 and 1.04% for FU and 0.52 and 0.59% for CPA) than those of manual preparation (respective values were 1.20 and 1.46% for FU and 1.70 and 2.20% for CPA). Drug residue was not detected from any of the prepared infusion bags with the robotic preparation, whereas FU was detected in two of four analyzed infusion bags with manual preparation. Average total time to make single anticancer drug preparation (compounding plus reconstitution of lyophilized drugs) was 6.11 min in the second half of 2015. During the study period, the highest percentage of production covered by APOTECAchemo was 70.4% of the total inpatient pharmacy activity.ConclusionRobotic preparation using APOTECAchemo should give substantial advantages in drug compounding for accuracy and safety and was able to be successfully worked in Mie university hospital.

Knowledge and Practices Regarding Usage of Biological Safety Cabinets

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 BSC is therefore important. This study evaluated BSC usage knowledge and practices and the effect of prior experience on training outcomes. Thirteen participants randomly selected from research centers were evaluated through a structured questionnaire covering biosafety considerations involved in using a BSC as per the World Health Organization’s Laboratory Biosafety Manual, third edition. The questionnaire was administered before and after the training. Training was conducted using the NUITM-KEMRI biosafety training program, enhanced to include more modules on BSCs. Differences in the participants’ knowledge and practices regarding BSC usage were observed. After training, the participants were able to give correct responses. Furthermore, participants who had fewer years of experience showed the greatest improvement. Results indicate the need for proper training of laboratory workers early in their careers, coupled with close supervision, mentorship, and regular refresher training.

Comparison of respiratory inductive plethysmography versus head-out plethysmography for anesthetized nonhuman primates in an animal biosafety level 4 facility

For inhalational studies and aerosol exposures to viruses, head-out plethysmography acquisition has been traditionally used for the determination of estimated inhaled dose in anesthetized nonhuman primates prior to or during an aerosol exposure. A pressure drop across a pneumotachograph is measured within a sealed chamber during inspiration/exhalation of the nonhuman primate, generating respiratory values and breathing frequencies. Due to the fluctuation of depth of anesthesia, pre-exposure respiratory values can be variable, leading to less precise and accurate dosing calculations downstream. Although an anesthesia infusion pump may help stabilize the depth of sedation, pumps are difficult to use within a sealed head-out plethysmography chamber. Real-time, head-out plethysmography acquisition could increase precision and accuracy of the measurements, but the bulky equipment needed for head-out plethysmography precludes real-time use inside a Class III biological safety cabinet, where most aerosol exposures occur. However, the respiratory inductive plethysmography (RIP) acquisition method measures the same respiratory parameters by detecting movement of the chest and abdomen during breathing using two elastic bands within the Class III biological safety cabinet. As respiratory values are relayed to a computer for software integration and analysis real-time, adjustment of aerosol exposure duration is based on the depth of sedation of the animal. The objective of this study was to compare values obtained using two methodologies (pre-exposure head-out plethysmography and real-time RIP). Transitioning to RIP technology with real-time acquisition provides more consistent, precise, and accurate aerosol dosing by reducing reported errors in respiratory values from anesthesia variability when using pre-exposure head-out plethysmography acquisition.

Laboratory biological safety cabinet (BSC) explosion

Scientists at universities across Iraq are actively working to report actual incidents and accidents occurring in their laboratories in order to raise awareness and encourage openness, leading to widespread adoption of robust Chemical Safety and Security (CSS) practices. In this study, we describe a serious event that resulted in a postgraduate student sustaining serious injuries when the biological safety cabinet (BSC) she was using exploded. Of particular note, the paper highlights how a combination of failures and deficiencies at many levels within an organization and its technical community (rather than a single piece of faulty equipment or the careless behaviour of one person) can lead to a dangerous, potentially life-threatening incident. By openly sharing what happened along with the lessons learnt from the accident, we hope to minimize the possibility of another researcher being injured in a similar incident in the future.

Evaluation of Environmental Cytotoxic Drug Contamination in a Clinical Setting.

The use of cytotoxic drugs to treat neoplastic conditions is increasing in veterinary practices. Many agents have the potential to be genotoxic and evidence of exposure to cytotoxic drugs has been found in healthcare workers handling these pharmaceuticals. To date, limited contamination evaluations have been performed in veterinary practices. Evaluation for carboplatin contamination was performed at a veterinary teaching hospital involving twelve areas in the dispensary and treatment room. Detectable levels of platinum were found on the surface of the biological safety cabinet where drugs are transferred from vial to administration device. Results indicate contamination did occur and care must be taken during all phases of cytotoxic drug preparation and administration; precautionary procedures must be in place to limit the spread of surface contamination and personnel exposure.

Effectiveness of activated carbon masks in preventing anticancer drug inhalation.

BackgroundThe exposure of healthcare workers to anticancer drugs such as cyclophosphamide (CPA) is a serious health concern. Anticancer drug pollution may spread outside biological safety cabinets even when a closed system is used. The inhalation of vaporized anticancer drugs is thought to be the primary route of exposure. Therefore, it is important that healthcare workers wear masks to prevent inhalation of anticancer drugs. However, the permeability of medical masks to vaporized anticancer drugs has not been examined. Furthermore, the performance differences between masks including activated carbon with chemical adsorptivity and non-activated carbon masks are uncertain. We investigated activated carbon mask permeability to vaporized CPA, and assessed whether inhibition of vaporized CPA permeability was attributable to the masks’ adsorption abilities.MethodsA CPA solution (4 mg) was vaporized in a chamber and passed through three types of masks: Pleated-type cotton mask (PCM), pleated-type activated carbon mask (PAM), and stereoscopic-type activated carbon mask (SAM); the flow rate was 1.0 L/min for 1 h. The air was then recovered in 50 % ethanol. CPA quantities in the solution were determined by liquid chromatography time-of-flight mass spectrometry. To determine CPA adsorption by the mask, 5 cm2 of each mask was immersed in 10 mL of CPA solution (50–2500 μg/mL) for 1 h. CPA concentrations were measured by high-performance liquid chromatography with ultraviolet detection.ResultsFor the control (no mask), 3.735 ± 0.543 μg of CPA was recovered from the aerated solution. Significantly lower quantities were recovered from PCM (0.538 ± 0.098 μg) and PAM (0.236 ± 0.193 μg) (p < 0.001 and p < 0.001 vs control, respectively). CPA quantities recovered from all of SAM samples were below the quantification limit. When a piece of the SAM was immersed in the CPA solution, a marked decrease to less than 3.1 % of the initial CPA concentration was observed.ConclusionThe SAM exhibited good adsorption ability, and this characteristic may contribute to avoiding inhalation exposure to vaporized CPA. These results suggest that wearing activated carbon masks may prevent anticancer drug inhalation by healthcare workers.

A Biological Safety Cabinet Certification Program: Experiences in Southeast Asia.

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.

Making the Most from Changes in Life

Making the Most from Changes in Life