Under standard Workplace Health & Safety guidelines, prior to entering the lab for the first time, team members were given a safety induction. This outlined both appropriate laboratory behaviour and laboratory safety features such as: the location of emergency showers; eyewash stations; fire and chemical extinguishers; fire blankets; and emergency power shut off points. All team members wore appropriate personal protective equipment upon entry into the laboratory, which consisted of a lab coat, enclosed sturdy footwear, safety glasses, and tied back hair (where applicable). No eating or drinking occurred within the laboratory and all wet lab work was conducted within the laboratory.

A laboratory technician,academic advisor, or staff member, was always present when any wet lab work was being conducted within the laboratory, and all team members were briefed on the operation of certain pieces of equipment prior to their usage. All potentially hazardous biological substances was disposed of into relevant waste and biohazardous material bins, and benches were cleaned down with ethanol to disinfect them at the end of the day, or after the completion of our experiments. We used designated fridges and freezers for storing our biological products and used biosafety cabinets when diluting H₂ gas producing, Hydrogen Gas Production Gene Cluster cultures down to a concentration required for Clarke electrode measurements.

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The E. coli K-12 strain, DH5α, we have modified (or engineered) is classified under Australian Law as a safe strain as host genes cannot be transferred and is unlikely to mutate. The E.coli is a knockout strain from the human gut microbiome and would require additional nutrients to grow in the environment freely (the addition of thiamine and leucine), and it has no pathogenesis factors classifying it as a biosafety level 1 organism (BSL-1).

The genes we have added are from Chlamydomonas reinhardtii and are therefore not readily found throughout environment in plants, algae, and other bacteria. These genes would not give rise to a resistance or survival advantage, especially as they have been engineered to produce hydrogen gas, a highly unlikely advantageous trait. These traits are more likely to be disadvantageous and impart the organism with reduced fitness as its metabolism has been re-engineered to produce non-essential, and possibly growth inhibitory, output.

Horizontal transfer could occur in the environment even if our Hydrogen Gas Production Gene Cluster cells were dead as the plasmid may still be transferred, however as mentioned, the plasmid does not contain any advantageous genes and therefore would not pose environmental risk.

Working in an Australian laboratory places us under the Gene Technology Act 2000. Therefore we have to protect the health, safety of individuals, and of the environment. We have done so by identifying the risks posed by, or as a result of our technology and have managed these risks by regulating our dealings with our technology. We have been given approval by the Macquarie University Institutional Biosafety Committee to work with the K-12 based Escherichia coli strain, which under the Gene Technology Act 2000 is classified as an exempt dealing. And we therefore will not release it into the environment. Overall the project is still in its design phase which takes place within the lab environment. If it went to market, it would have to be trialled in a consultation phase, controlled release and then a final phase release. All of our work complied with the Office of the Gene Technology Regulator, Australia’s national regulatory body.

Australian Government, Department of Health – Office of the Gene Technology Regulator. Gene Technology Act 2000. (Commonwealth).