Applied Biosafety

With increased research efforts on vector-borne diseases such as malaria, dengue, and recently Zika, there has been a need to maintain in captivity arthropods that are known or believed to be involved in the transmission of pathogens to humans in a well-controlled laboratory environment. Therefore, it is pivotal to establish safe yet practical procedures inside an insectary to minimize the associated biosafety risks and prevent the potential escape of the arthropods used into the environment. This article provides a detailed overview of an arthropod disease vectors facility features with a focus on arthropod containment level 2 (ACL-2) laboratory and aims to support health safety and environment (HSE) and biosafety professionals as well as scientists who work on vector-borne diseases by detailing safety procedures inside an ACL-2 environment. In particular, I detail here the associated safety procedures of an ACL-2 and the insectary-specific equipment required for this type of facility. Research on mosquitoes was used here as an example of a disease vector and presents procedures relevant to usage of this vector in the context of Zika virus. The work presented here shows that risks associated with working on infected arthropod vectors can be minimized to an acceptable level and in turn facilitate the research on the diseases they help spread. Similar principles can be applied to other vector-borne diseases research as well, although the work may have a distinct nature.

Emerging technologies in the life sciences call for new models of biosafety risk management. We examine the question of how to address new developments in the life sciences and biosciences in a bottom-up manner—that is, from the concrete level of biosafety practice with a focus on the risk management and risk assessment of emerging technologies in the biology laboratory. We use research on “gene drives” as an example of challenging work with new constructs that have major biosafety implications for the work in the laboratory and beyond. Gene drives are intended for use in ecosystems and require, at an early stage, the consideration of potential future biosafety, biosecurity, and societal impact. We argue for an integrative approach, a truly collaborative model that involves scientists, biosafety officers, institutional leadership, and ethics consultants, with the aim of maximizing safety as well as scientific progress.

The ability to detect and quantify viral vector sequences from a variety of clinical sample types is crucial to performing biosafety risk assessments. Viral vector-mediated gene transfer studies are often performed in animals, and these animals must be placed under appropriate biocontainment conditions to protect the workers and environment. Data on the shedding of viral vectors from animals are limited, and the sample types are challenging for polymerase chain reaction (PCR). For this reason, we developed a quantitative PCR assay that could be used for such purpose. We designed a sequence-specific, probe-binding method for detecting adenoviral and lentiviral vector sequences. A duplex strategy was used that included a quality control sequence to be amplified in the same reaction as the viral vector. This sequence provided an internal control for normalization of noncellular sample types, such as animal excretions that inherently lack a natural control sequence. The new assay was used to establish the efficiency of reverse transcription and to detect viral genomes in stocks of whole virus particles. We identified sets of primers and probes for both adenoviral and lentiviral sequences that work well together with no interference. The average conversion rate of RNA into complementary DNA was 18.5%. The new quantitative PCR assay was efficient and specific, and it measured successfully the number of viral genomes in stocks of whole virus particles. This assay could be used to detect adenoviral and lentiviral vector sequences for biosafety and other research purposes.

The objective of this study was to evaluate various procedures for decontamination of vehicles and equipment during outbreaks of notifiable animal diseases in cold weather. The evaluation was done in 4 field trials held in outdoor operational settings in Canada, at ambient temperatures from –2°C to 11°C. Procedures included various combinations of dry cleaning, wet cleaning, disinfection, and final rinsing. Geobacillus stearothermophilus spores and infectious bursal disease virus (IBDV) were used as surrogates for bacterial and viral pathogens, particularly Bacillus anthracis spores and foot-and-mouth disease virus. Spores and viruses were suspended in a light organic soil preparation, inoculated onto stainless-steel disks, and covered with a heavy soil preparation. Inoculated disks were attached to various surfaces of farm vehicles and equipment. In all field trials, spore and IBDV reduction was greater (P < .05) on disks where the soil was completely removed, as compared with only partially removed. Greater (P < .05) spore and IBDV reduction was seen when disinfection and final rinse steps were included than when not included after dry and wet cleaning in 3 of 3 and 1 of 3 field trials, respectively. A wet-cleaning step before application of a disinfectant increased (P < .05) spore and IBDV reduction versus no wet cleaning. The results provide evidence that vehicle and equipment decontamination in cold weather could be significantly improved by thorough removal of organic matter to enhance disinfection and elimination of disease agents.

Academic medical centers comprise a diverse group of students, researchers, and health care professionals. There are also 3 unique environments: education, health care, and research. Differences in biosafety practices and procedures exist between these environments, but the common goal is to produce well-trained and successful health care and research professionals. An important part of the educational experience should include biosafety training. Having a strong foundation on biosafety principles will increase safety compliance in both the clinical and research workforces. Both students and employees should be knowledgeable and aware of biosafety. A biosafety survey was developed to measure the knowledge of students on biosafety principles at an academic medical center. This survey was validated by field professionals, including physicians, nurses, laboratory technologists, and researchers, who were surrogates for a multidisciplinary student population. The goal of the validation was to determine if the biosafety survey included appropriate questions and responses to adequately assess biosafety knowledge among students at an academic medical center. This step was very important because there are differences in the practices and guidelines for health care and research arenas. The validation results indicated that there are differences in the understanding of biosafety principles among different disciplines. There are also major differences in interpretations of biosafety principles between clinicians and researchers.