This Guideline is aimed specifically at Canadian facilities handling human and terrestrial pathogens or toxins, and elaborates on key topics introduced in the Canadian Biosafety Handbook (CBH) to serve as a resource for stakeholders seeking additional information to establish a robust biosecurity plan. In Canada, most facilities where human and terrestrial animal pathogens or toxins are handled and stored are regulated by the Public Health Agency of Canada (PHAC) and the Canadian Food Inspection Agency (CFIA) under the _Human Pathogens and Toxins Act _(HPTA), _Human Pathogens and Toxins Regulations (HPTR), Health of Animals Act (HAA), and Health of Animals Regulations _(HAR). Regulated facilities must develop and maintain a biosecurity plan in accordance with requirements set forth in the Canadian Biosafety Standard (CBS), 2nd edition. These Canadian Biosafety Guidelines have been developed by the PHAC and CFIA as part of an ongoing series of biosafety and biosecurity themed guidance documents. The Public Health Agency of Canada focuses on preventing disease and injuries, responding to public health threats, promoting good physical and mental health, and providing information to support informed decision making. The CFIA is charged with safeguarding, food, animals and plants in order to enhance the health of Canada’s people, environment, and economy. While these guidelines are specifically designed for facilities to meet Canadian regulatory requirements, they may be used as an example of biosafety guidelines for other nations and non-Canadian laboratories. This document is publicly available as a PDF in both English and French.