Abstract

Climate change and global movements of people and goods have accelerated the spread of invasive species, including insects that vector infectious diseases, which threaten the health of more than half of the world’s population. Increasing research efforts to control these diseases include the study of vector – pathogen interactions, involving the handling of pathogen-infected vector insects under biosafety level (BSL) 2 and 3 conditions. Like microbiology BSL-3 laboratories, BSL-3 insectaries are usually subjected to fixed-term or emergency room decontamination using recognized methods such as hydrogen peroxide (H2O2) or formaldehyde fumigation. While these procedures have been standardized and approved for the inactivation of diverse pathogens on surfaces, to date, there are no current standards for effective room-wide inactivation of insects in BSL-3 facilities in case of an emergency such as the accidental release of a large number of infected vectors. As H2O2 is often used for standard room decontamination in BSL-3 facilities, we evaluated H2O2 fumigation as a potential standard method for the safe, room-wide deactivation of insects in BSL-3 insectaries in comparison to heat treatment. To account for physiological diversity in vector insect species, six species from three different orders were tested. For the H2O2 fumigation we observed a strong but also varying resilience across all species. Lethal exposure time for the tested dipterans ranged from nine to more than 24 h. Furthermore, the coleopteran, Tribolium castaneum, did not respond to continuous H2O2 exposure for 48 h under standard room decontamination conditions. In contrast, temperatures of 50°C effectively killed all the tested species within 2 to 10 min. The response to lower temperatures (40–48°C) again showed a strong variation between species. In summary, results suggest that H2O2 fumigation, especially in cases where a gas generator is part of the laboratory equipment, may be used for the inactivation of selected species but is not suitable as a general emergency insect inactivation method under normal room decontamination conditions. In contrast, heat treatment at 48 to 50°C has the potential to be developed as an approved standard procedure for the effective inactivation of insects in BSL-3 facilities.

Highlights

  • The increasing worldwide threat by emerging and re-emerging vector-borne infectious diseases has boosted related research activities to study vector-pathogen interactions and enable the development of diagnostic tests, vaccines, and drugs

  • Blackborough, Imperial College, London, United Kingdom), Drosophila melanogaster [Oregon-R strain (Lindsley and Grell, 1968), adults, mixed-sex], and Ceratitis capitata representing the Diptera; (2) Tribolium castaneum (San Bernardino laboratory strain, adults, mixed-sex; strain sourced from Fraunhofer IME-Bioresources) representing the Coleoptera; and (3) Spilostethus pandurus representing the Hemiptera

  • We tested if either H2O2 fumigation or heat treatment has the potential to be developed into a standard procedure for the safe and ethical room-wide decontamination of biosafety levels (BSL)-3 insectaries in emergency situations, such as mass escapes of pathogen-infected insects

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Summary

Introduction

The increasing worldwide threat by emerging and re-emerging vector-borne infectious diseases has boosted related research activities to study vector-pathogen interactions and enable the development of diagnostic tests, vaccines, and drugs. Research involving vector-borne diseases often involves experiments on pathogen-infected insects, requiring specific containment and decontamination measures, which depend on the biosafety classification of the pathogen. There are four globally recognized biosafety levels (BSL) defined by the Centers for Disease Control and Prevention in the United States that have been adopted in Europe under Directive 90/679/EEC (Council of the European Union, 1990). The lowest biosafety level, BSL-1, is appropriate when there is little danger to personnel or the environment. The highest level (BSL-4) is required for transmitted pathogens that cause severe to fatal human diseases for which there are no available vaccines or treatments

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